CLPRST1

Clinical Parasitology
Module
For the Use of University of Baguio School of Natural
Sciences students only
Compiled and Prepared by

Jaleh V. Gacayan, RMT, LPT, MPH
Jessa Felix, RMT
Erlinda P. Sanchez, RMT, MPA,LIB
A Self-regulated Learning Module 2
INTRODUCTION
Clinical parasitology is an area of biology concerned specifically to the
study of important parasites which causes diseases to man. It covers the
classifications of parasites, symptoms, diseases caused by the parasites,
lifecycle, transmission, and treatment. The diseases caused by these parasites
constitute major human health problems throughout the world.
Parasitology is an essential component of clinical laboratory medicine.

The results obtained through proper specimen examination of parasites will
provide crucial information with regards to the diagnosis and treatment of
human disease. Tracking the epidemiology of such organisms as well as
establishing preventive mechanisms may be accomplished with the assistance
of this information.
In spite of the many advances in technology that has been developed in

recent years, the conventional technique of manually processing and
examining the samples both macroscopically and microscopically still occurs in
select clinical settings. It is essential that well-educated and extremely trained
individuals perform these procedures as well as read and interpret the results.
Thus, the goal of this module is to provide the needed information for students
preparing for a career in medical laboratory science. The module is designed to
help the learners in both the lecture and laboratory components related with
clinical parasitology. Students using this module will have the opportunity to
advance their skills necessary to become proficient laboratory scientists.

COURSE TITLE AND DESCRIPTION
This module is a learning package for the CLPRST 1-Clinical Parasitology
offered as a professional course in Bachelor of Science in Medical Laboratory
Science. It comprises of three (3) units lecture and one (1) unit laboratory.
This course deals with the study of human parasites which are of medical
importance especially those commonly found in the Philippines. Emphasis is
given on the epidemiology, pathogenecity, distribution, life cycle, and
laboratory identification of each parasite. Preventive measures against infection
and control are also given emphasis.
ABOUT THE MODULE

This module was designed to adopt the blended learning approach in
teaching Clinical Parasitology to the BSMLS students via online and distance
learning.
Learning Engagement1 of the module deals with the overview of what
Clinical Parasitology is. It includes an introduction which covers the terminologies
commonly used in the course, the different ways of specimen processing to
isolate a parasite, Techniques Used in the Identification of Parasites as well as
prevention and control. It also covers the morphology, pathophysiology, life
cycle, specimens used for the identification, diagnostic features, prevention and
control of Nematodes (Roundworms).
Learning Engagement 2 of the module is about the Morphology,
Pathophysiology, Life cycle, Specimens used for identification, Diagnostic
features, Prevention and Control of Cestodes (Tapeworms). It also deals with
the Morphology, Pathophysiology, Life cycle, Specimens used for identification,
Diagnostic features, Prevention and Control of Trematodes (Flukes).
Learning Engagement 3 covers the Morphology, Pathophysiology, Life
cycle, Specimens used for identification, Diagnostic features, Prevention and

Control of Protozoa. It also deals with the characteristics and role of
ectoparasites affecting man.
COURSE REQUIREMENTS
The pre-requisite assigned for the course is Human Anatomy and
Physiology.
Student’s attendance in the online learning sessions is a basic requirement
in which university policy on tardiness and absences are applied.
Various assessment tools are employed in the course to gauge the
student’s level of understanding and comprehension within the duration of the
online learning sessions throughout the term.
It is hoped that by the end of the course, students have instilled the
necessary and essential skills in clinical parasitology that would equip them in
their practice of the profession.

Requirements of the Course
1. Regular Attendance to classes: You must attend online classes and live quizzes
regularly by logging in to our scheduled online activities. Online lectures will be
done through Google meet and/or facebook live. Assessments shall be given
through Quizziz, Pear Deck, Canvas and/or Google forms. For offline students,
your attendance will be monitored through your responses to text information
and through timely correspondence.
2. Submission of required activities: All required activities (assignments, research
work, laboratory illustrations) should be submitted on or before the given
deadline. Deadlines will be posted by the teacher in the google classroom and
messenger group chat. It will also be texted to offline students. For online
students, requirements must be submitted to the teacher’s email address which
will be provided during the class orientation. For offline students, requirements
must be submitted via mail or express courier (e.g. LBC, JRS) addressed to:
Instructor’s name, School of Natural Sciences, University of Baguio, Baguio City.
3. Seventy percent (70%) passing score in all required activities: Quizzes, exams,
assignments, research work, laboratory illustrations.
Computation of grades:
▪ The Course Grade is obtained by combining the lecture and laboratory
grades (50%:50%) for the subject.
▪ Laboratory grade shall be computed as 30% enhancement activities
(illustrations; research work; case study; experiments – when possible) plus 70%
class standing (quizzes and exams).
▪ The cumulative system of computing grades shall be followed. Grades
computed for midterms and finals are considered tentative. The final midterm
grade is calculated by getting 1/3 of the first grading grade plus 2/3 of the
tentative midterm grade and the final grade is computed by getting 1/3 of
the midterm grade plus 2/3 of the tentative final grade.
4. Study/Learning Guidelines:
a. Manage your time properly. As students of higher education (College), you
are expected to be more responsible in paying attention to course schedules,
requirements, and deadlines. Schedule how you will accomplish all the
requirements in all your enrolled courses (reading the modules, reading on
research/ enhancement questions, doing assignments and laboratory
illustrations) and focus your attention when doing your tasks.
b. Observe proper conduct. Despite this online mode of learning, you must still
maintain appropriate behavior at all times. All standards of student conduct
outlined in the University of Baguio Student Handbook remain in full effect
during this time of distance learning. Be honest in answering your quizzes and
exams. Work independently when accomplishing tasks and assignments.

c. Stay motivated. Your future depends on what you do today. Maintain a
positive attitude towards learning and enjoy a fun-learning environment
despite the current circumstances.
d. Maintain a performance of high standard. Give your best in accomplishing all
the assigned tasks. Do not be complacent with just a 70% passing cut-off
score. Remember that this is a board subject, and the best preparation for the
board/licensure examination should be during these formative years. The
board review is but supplementary to the knowledge you have already
learned during your Med Tech education.
e. Communicate properly. Promptly respond to notifications by regularly visiting
our google classroom and messenger group chat. If you have confusions or
queries in any part of this module, I am here to guide you through. Send your
academic concerns using the same online platforms. For offline students, text
messages and mobile calls are welcome during scheduled hours of the day
and week. Be guided by this schedule when communicating:
▪ Respect private hours. I do not always open my laptop/email/messenger
24/7. Send your queries and/or concerns during regular office hours. For
concerns that need immediate attention, send through mobile text.
▪ Be patient. Messages received between 8 AM to 8 PM will be responded
to within the same day. Messages received after 8 PM will be answered
starting 8 AM the next day.
▪ Before calling my mobile number, text first for permission for I might be
giving an online lecture or in a meeting or on private moment at that
very instance.
▪ Saturdays and Sundays are for my family and home chores. I shall
respond to queries/messages received during these days within the first
office hour of Monday.
f. Show mutual support. Support one another. Let us all be responsible and
supportive in making this new learning process more effective.
g. Live lecture/Video conferencing guidelines:
g.1 Be punctual. Live lectures/Video conferences will be scheduled during the
official class period/time of this course. Log in to the platform at least 5-10
minutes before the class period. Prepare your learning materials such as
this module, pens, papers, etc. Attendance will be checked during the
lecture/video conference.
g.2 Maintain professionalism.
- Wear appropriate clothing and set your gadget in an appropriate
area. You may be asked to turn on your video/camera at any time
during the lecture.

- Log in using your UB gmail account. Unidentified names like nicknames,
phone models, etc. will not be allowed in the video conference.
- Mute your microphone as soon as you log in to the platform to avoid
any excess background noise. Unmute your microphone when
instructed to do so.
- Be courteous. Do not interrupt your teacher or a classmate who is
speaking. You may type your question in the Chat area, or use the
“raise hand” feature if available, and wait until you are allowed to
speak.
- Respect privacy. Do not take a screenshot, picture, snapshott, etc. of
your teacher or fellow students, nor make any unnecessary audio or
video recordings.
g.3 Remain focused and engaged. Do not be distracted by your gadget.
Keep your videoconference platform open and do not navigate other
tabs or webpages unless directed by your teacher.
Endorsed by:
Teresa N. Villanueva, RMT, MACT

Dean, School of Natural Sciences

STUDY SCHEDULE
WEEK TOPIC ACTIVITY
1 Section 1 Introduction to Online Lecture
Parasitology Laboratory : Experiment 1 & 2
2 Continue Section 1 Introduction to Online Lecture, Assessment & Quiz

Parasitology Laboratory : Experiment 3 and 4
3 Section 2: Nematose Intro Online Lecture: Assessment & Quiz

Section 2A Adenophorea Laboratory: Experiment 5
4 Section 2B: Phasmidea Online Lecture: Assessment & Quiz
Laboratory: Experiment 6 and 7
5 Section 2B: Phasmidea Online Lecture: Assessment & Quiz
Blood and Tissue Nematodes Laboratory: Experiment 8 and 9
6 FIRST GRADING EXAM Coverage Section 1 and 2
7 Section 3: Cestodes Online Lecture: Assessment & Quiz
- Introduction Laboratory: Experiment 10
8 Section 3: Cestodes Online Lecture: Assessment & Quiz
Laboratory: PRACTICALS
9 Section 4: Trematodes Online Lecture: Assessment & Quiz
- Introduction Laboratory: Experiment 11
10 Section 4A: Dioecious Flukes Online Lecture: Assessment & Quiz
Laboratory: Experiment 12
11 Section 4B: Monoecious Flukes Online Lecture: Assessment & Quiz
Laboratory: PRACTICALS
12 MIDTERM Coverage Lesson 1-10
13 Section 5A: Sarcodina (Amoeba) Online Lecture: Assessment & Quiz
14 Section 5B: Mastigophora Online Lecture: Assessment & Quiz
(Flagellates) Laboratory: PRACTCALS
15 Section 5C: Mastigophora Online Lecture: Assessment & Quiz
(Hemoflagellates) Laboratory: Experiment 14
16 Section 5D: Ciliophora (Ciliates) Online Lecture: Assessment & Quiz
17 Section 5E: Apicomplexa (Sporozoa) Online Lecture: Assessment & Quiz
18 Section 6: Ectoparasites Online Lecture: Assessment & Quiz
Laboratory: EXAMINATION
19 FINALS Coverage Lesson 1-17

Table of Contents
Section 1: Introduction to Parasitology ............................................................................................. 11
Experiment no. 1: Instrument and Glasswares ............................................................................ 37
Experiment no. 2: Physiology of Stool Formation and Specimen Collection ............................... 40
Experiment no. 3: Routine Stool Examination (Physical and Chemical Phase) ............................. 43
Experiment no. 4: Routine Stool Examination (Microscopic) ...................................................... 46
Section 2: Nematodes ..................................................................................................................... 52
Section 2A: Adenophorea ................................................................................................................... 56
Experiment no. 5: Phylum Nematoda (Aphasmidea) ................................................................. 67
Section 2B: Phasmidea ....................................................................................................................... 72
Experiment no. 6: Phylum Nematoda (Phasmidea) .................................................................. 103
Experiment no. 7: Phylum Nematoda (Secernentia; Phasmidea) ............................................. 109
Experiment no. 8: Phylum Nematoda (Blood and Tissue Nematodes) ..................................... 117
Experiment no. 9: Phylum Nematoda (Other Members) ......................................................... 122
Section 3: Cestodes ....................................................................................................................... 125
Experiment no. 10: Phylum Platyhelminthes (Class Cestoda) .................................................. 145
Section 4: Trematodes ................................................................................................................... 150
Section 4A: Dioecious Flukes ........................................................................................................... 154
Experiment no. 11: Phylum Platyhelminthes (Class Trematoda – Dioecious flukes) .................. 163
Section 4B: Monoecious Flukes ....................................................................................................... 167
Experiment no. 12: Phylum Platyhelminthes (Class Trematoda – Monoecious flukes).............. 199
Section 5: Protozoa ....................................................................................................................... 204
Section 5A: Sarcodina (Amoeba) ...................................................................................................... 209
Experiment no. 13: Phylum Sarcomastigophora (Subphylum Sarcodina) .................................. 233
Section 5B: Mastigophora (Flagellates) ............................................................................................ 237
Section 5C: Mastigophora (Hemoflagellates) ................................................................................... 246
Experiment no. 14: Phylum Sarcomastigophora (Subphylum Mastigophora) ........................... 256
Section 5D: Ciliophora (Ciliates) ....................................................................................................... 262
Experiment no. 15: Phylum Ciliophora....................................................................................... 265
Section 5E: Apicomplexa (Sporozoa) ................................................................................................ 268
Experiment no. 16: Phylum Apicomplexa (Class Sporozoa) ....................................................... 284
Section 6: Ectoparasites ......................................................................................................................... 289

LEARNING ENGAGEMENT 1 CLINICAL PARASITOLOGY: OVERVIEW
INTRODUCTION:
Parasitology is the area of biology concerned with the phenomenon of
dependence of one living organism to another. Medical Parasitology is
concerned primarily with the animal parasites of humans and their medical
significance, as well as their importance in human communities. It includes the
study of three major groups of animals: parasitic protozoa, parasitic helminths
(worms), and the arthropods that directly cause disease or act as vectors of
different pathogens. A parasite is a pathogen that simultaneously injures and
derives sustenance from its host. Some organisms called parasites are actually
commensals, in that they neither benefit nor harm their host (for example,
Entamoeba coli). Although parasitology had its origins in the zoologic sciences,
it is today an interdisciplinary field, greatly influenced by microbiology,
immunology, biochemistry, and other life sciences. The basic biology of the
pathogens, their host-parasite relationships and descriptions of the basic
properties of the pathogens, the pathogenesis of the diseases they cause, host
defenses, and epidemiology are highlighted in this module.

SECTION 1: INTRODUCTION TO PARASITOLOGY
COURSE LEARNING OUTCOMES:
At the end of this session, the students must have:
1. used the basic terminologies and theories related to Parasitology

2. enumerated the precautions and guidelines to be considered in proper
specimen collection
3. appreciated the significance of laboratory waste management, prevention and
control of parasitic infections, and the maintenance of a safe laboratory
environment
4. established guidelines on acceptability of stool samples in the laboratory
5. interpreted and decided on the negativity or positivity of the sample studied and
follow the correct manner of reporting
LESSON PROPER
I. TERMINOLOGIES
PRE-ACTIVITY: Find the meaning of the following terminologies and indicate the
reference used for each.
1. Parasitology 11. Temporary parasite

2. Parasitism 12. Permanent parasite
3. Symbiosis 13. Pathogenic parasite
4. Commensalism 14. Pseudoparasite
5. Mutualism 15. Coprozoic/spurious parasite
6. Parasite 16. Host
a) Ectoparasite 17. Final/definitive host
b) Endoparasite 18. Intermediate host
7. Facultative parasite 19. Accidental host
8. Obligate parasite 20. Paratenic host
9. Erratic parasite 21. Dead-end host
10. Incidental parasite 22. Reservoir host

23. Parasitic infection 27. Oviparous female parasite
24. Parasitic infestation 28. Parthenogenic female parasite
25. Parasitic disease 29. Protandry
26. Larviparous/viviparous female
parasite
II. PARASITIC INFECTION
A. PARASITE-HOST RELATIONSHIPS
Organisms may develop unique relationships due to their habitual and long
associations with one another. These relationships are very important to their survival.
Symbiosis is the living together of unlike organisms. It may involve protection or other
advantages to one or both partners. Different forms of symbiosis may be distinguished
on the basis of whether or not the association is detrimental to one of the two
partners. Commensalism is a symbiotic relationship in which two species live together
and one species benefits from the relationship without harming or benefiting the other.
Mutualism is a symbiosis in which two organisms mutually benefit from each other.
Parasitism is a symbiotic relationship where one organism, the parasite, live in or on
another, depending on the latter for its survival and usually, at the expense of the host.
Table 1-1 lists the terms associated with parasite-host relationships, along with their
definitions.

There are several types of parasites that may be members of a parasite-host
relationship. An organism may be an obligatory parasite or a facultative parasite. It
may be an endoparasite or an ectoparasite. In the same manner, a number of
different hosts may be part of this parasite host relationship. These include accidental
or incidental hosts, definitive hosts, intermediate hosts, reservoir hosts, transport hosts,
and carriers.
When a parasite infects a host, symbiosis results. The primary function of the host
is to carry on the parasite’s life cycle. This newly formed relationship may develop into
commensalism, mutualism, or parasitism. Some of these associations exist as
commensal under certain circumstances and pathogenic under others. Parasites
have an amazing capability to adapt to their host surroundings. In addition to a
number of morphologic adaptations, parasites
are capable of protecting themselves from the host’s immune system. Parasites alter
their antigenic makeup so that the host will not recognize the modified parasites as
foreign, and thus the initiation of an immune response does not
occur.
B. EXPOSURE and INFECTION
Majority of animal parasites are pathogens which are harmful and which
frequently cause mechanical injury to their hosts. A carrier harbors a particular
pathogen without manifesting any signs and symptoms.
Exposure is the process of inoculating and infective agent, while infection
connotes the establishment of the infective agent in the host.
The incubation period is the period between infection and evidence of
symptoms. It is sometimes referred to as the clinical incubation period. The pre-patent
period, also known as the biologic incubation period, is the period between infection
or acquisition of the parasite and evidence of demonstration of infection.
Autoinfection results when an infected individual becomes his own direct
source of infection. On the other hand, superinfection or hyper infection happens
when the already infected individual is further infected with the same species leading
to massive infection with the parasite.
C. SOURCES OF INFECTION

There are various sources of parasitic infections. The most common sources are
contaminated soil and water. Lack of sanitary toilets and the use of night soil or
human excreta as fertilizer allow the eggs to get in contact with the soil. Another
possible source of infection is water and food, which contain the infective stage of the
parasite. Consumption of undercooked or raw fresh water fish can result in several
intestinal and liver fluke infections. Arthropods can also transmit infection. Mosquitoes
are vectors of malaria and filaria parasites. Other animals, whether wild or
domesticated, may also harbor the parasite. Other sources include another person,
his beddings and clothing, the immediate environment he has contaminated, or even
one’s self.
D. MODES OF TRANSMISSION
An infectious agent may be transmitted from its natural reservoir to a
susceptible host in different ways. There are different classifications for modes of
transmission:
1. Direct – contact w/ an infected person or animal, directly from the source
to the susceptible host without involving an intermediate object
a) Droplet spread
b) Sexual intercourse
c) Kissing
d) Holding hands
e) Transplacental / Vertical - mother to fetus
2. Indirect
a) Ingestion of contaminated food & drink
b) Contact w/ contaminated soil
c) Bite of an infected arthropod (vector)
d) Through fomites
Since the most common source of parasitic infection is contaminated food and
water, the most likely portal of entry is the mouth. Majority of infections among
cestodes, trematodes, and intestinal protozoans are foodborne: Taenia solium, Taenia
saginata, and Diphyllobothrium latum from eating food harboring the infective larval
stages; Entamoeba histolytica and Giradia lamblia from drinking water contaminated

with cysts; and Clonorchis, Opistorchis and Haplorchis through ingesting raw or
improperly cooked freshwater fish containing the parasitic larvae.
Skin penetration is another route of transmission. Hookworms and Strongyloides
enter via exposure of skin to soil, while Schistosoma species enter skin via water.
Arthropods also serve as vectors and transmit parasites through their bites.
Examples are agents of malaria, filariasis, leishmaniasis and trypanosomiasis.
Another way of acquiring infection is through congenital transmission.
Toxoplasma gondii trophozoites can cross the placental barrier during pregnancy. In
transmammary infection with Ancylostoma and Strongyloides, the parasites may be
transmitted through the mother’s milk.
Other ways of acquiring the infection include inhalation of airborne eggs of
Enterobius, and sexual intercourse as in the case of Trichomonas vaginalis.
E. LIFE CYCLE
Although parasitic life cycles range from simple to complex, they all have three
common components—a mode of transmission, a morphologic form that invades
humans, known as the infective stage, and one (or more) forms that
can be detected via laboratory retrieval methods, known as the diagnostic stage.
Some parasites require only a definitive host, whereas others also
require one or more intermediate hosts. A parasitic life cycle consists of two common
phases (Fig. 1). One phase involves the route a parasite follows when in or on the
human body. This information provides an understanding of
the symptomatology and pathology of the parasite. Insights about the best the
method of diagnosis and selection of appropriate antiparasitic medication may also
be determined. The other phase, the route a parasite follows independently
of the human body, provides crucial information pertinent to epidemiology,
prevention, and control.

Figure 1. Generic Parasite life cycle
F. DISEASE PROCESSES and SYMPTOMS

A parasitic disease may affect the entire body or any of its parts. The major
body areas associated with such processes include the following: (1) the
gastrointestinal (GI) and urogenital (UG) tracts; (2) blood and tissue; (3) liver, lung, and
other major organs; and (4) miscellaneous locations, such as cerebrospinal fluid (CSF),
eye, skin, and extremities.
A wide variety of representative symptoms, may occur when a parasite

infects a human host. Some persons remain asymptomatic, whereas other parasites
produce severe symptoms and may result in death. The most commonly observed
symptoms include diarrhea, fever, chills, abdominal pain, and abdominal cramping.
Other symptoms, such as elephantiasis (an enlargement of areas such as the breast,
leg, and scrotum caused by a parasite’s presence), anemia, vitamin deficiency,
bowel obstruction, edema, enlargement of major organs, skin lesions, and blindness,
may develop.
G. PREVENTION and CONTROL

Prevention and control measures may be taken against every parasite infective
to humans. Preventive measures designed to break the transmission
cycle are crucial for successful parasite eradication. Examples of such measures
include the following: education programs, use of insecticides and other chemicals,
protective clothing, protective netting, proper water treatment, good personal
hygiene, proper sanitation practices, proper handling and preparation of food, and
avoidance of unprotected sexual relations. The vast capital expenditures required to
accomplish these measures are not available in many endemic countries in the world.
The problem of eradicating parasites is an
ongoing process and is a key goal of international health groups such as the World
Health Organization (WHO)
III. LABORATORY METHODS FOR DIAGNOSIS
A. SPECIMEN FOR DIAGNOSIS

The following are the specimens needed to properly diagnose the presence of
parasites inside the human body.
1. Stool- for intestinal protozoans, nematodes and helminthes
2. Urine- for the recovery of Trichomonas vaginalis and Schistosoma
haematobium
Collection: mid-stream catch
3. Sputum- Paragonimus westermani, larvae of nematodes
• Must be digested using 4-5% sodium hydroxide
4. Blood- for malarial parasites, filarial worms, Leishmania and Trypanosoma
5. Cerebrospinal fluid- Acanthamoeba species
• Collection: lumbar tap
6. Tissue aspirates:
a) Liver aspirate- hydatid cyst and liver amoebic abscess
b) Duodenal aspirate- Giardiasis and Strongyloidiasis infection
• Collection: endoscopy
• Duodenal drainage or “String test”: duodenal contents collected for
Giardia and Strongylodes
• Sigmoidoscopy: Schistosomiasis, Amoebiasis, Balantidiasis and Shigellosis
(Large intestines)
c) Broncho-alveolar lavage – Paragonimus westermani
7. Orifice swab
a) Vaginal swab- Trichomonas vaginalis
b) Perianal swab – Enterobius vermicularis and Taenia
8. Tissue Biopsy
a) Muscle- Trichinella spiralis
b) Rectal – granulomas secondary to Schistosomiasis
B. PROCESSING OF BLOOD SAMPLE
There are parasites where the method of isolation and identification is through
processing of blood.
❖ Blood Films
1. Fresh water smears- for diagnosis of Trypanosomes and microfilaria
2. Thin Dry smears – for the study of the morphology of the parasites and the blood
cells
3. Thick Dry smears – used for malaria survey among patients with chronic infections
or who are undergoing anti-malaria therapy.
❖ Stains
1. Giemsa – most preferred
Composition: Stock solution 1 ml
Buffered water (pH 7.0 – 7.2) 49 ml
Staining time: 30 minutes
- Too dark : acidic pH
- Too red: alkaline pH
2. Rapid stains:
a) Wright’s stain – It is used to stain blood smears in the detection of blood
parasites. The stain distinguishes easily between blood cells and became
widely used for performing differential white blood cell counts, which
are routinely ordered when infections are expected. The stain contains a
fixative, methanol, and the stain in one solution. Thin films of blood are
fixed with methanol to preserve the red cell morphology so that the
relationship between parasites to the red cells can be seen clearly.
✓ Fix with 1-2 drops of methanol
✓ Cover the film with 10% Giemsa stain: 5 minutes
✓ wash with distilled water, drain, dry and examine
b) Leishmann stain – It is a mixture of Methylene blue, and Eosin dye, prepared in
Alcohol medium and diluted with buffer or distilled water during staining
procedure. Leishman stain is a differential stain that is used to variably stain the
various components of the cells and it can be used to study the adherence of
pathogenic bacteria to the human cells. It differentially stains the human and
bacterial cells and appeared as purple and pink colored bodies respectively.
The Leishman stain is one of the best stains for routine blood stain to stain the
Peripheral blood smear for the examinations of blood film under the
microscope and is satisfactory for malaria and other blood parasites
✓ Add 7-8 drops of the stain: 1-2 minutes
✓ Add 12-15 drops of buffered distilled water
✓ Mix thoroughly, let stand (4-8 minutes)
✓ Rinse, drain, dry and examine
c) Field’s stain - It is a histological method for staining of blood smears. It is used for
staining thick blood films in order to discover malarial parasites. Field's stain
consists of two parts - Field's stain A is methylene blue and Azure 1 dissolved in
phosphate buffer solution; Field's stain B is Eosin Y in buffer solution.
d) Acridine orange - Acridine Orange Stain is used as a fluorescent staining agent
to detect the presence of malaria parasite in blood cultures and other bodily

fluids. Acridine orange is a fluorochrome dye that can interchalate into nucleic
acid.
e) Jaswant Singh Battacharya (JSB) Stain for thick and thin films - standard
method: laboratories under the National Malaria Eradication Programme in
India
3. Permanent stains & Other stains
a) Iron Hematoxylin Stain - used for most of the original morphological descriptions
of intestinal protozoa found in humans.
b) MIF Fixative Stain (Merthiolate iodine formaldehyde) - diagnosis of
Trichomonas
c) Chlorazol Black E - An acid dye, used as a fat and general tissue stain, and to
stain protozoa in fecal smears or in tissues.
d) Modified Kohn’s - modification of the chlorazol black E staining technique
e) Wheatley Trichrome - all-purpose (amoebae, flagellates)
f) Methenamine Silver - cyst
g) Fluorescent Staining- Microsporidium
PARASITEMIA
Thick Blood Film:
✓ Infected erythrocytes are counted in relation to a predetermined number of

WBCs
✓ Standard: average of 8000/µl
✓ All parasite species and forms (sexual and asexual) are counted
Formula:
# of parasites/µl = # of parasites x 8000

# of WBCs counted
• If parasites are ≥10, 200 leukocytes are counted:
# of parasites/µl =# of parasites counted x 40
• If parasites are <10, 500 WBCs should be counted:
# of parasites/µl = # of parasites counted x 16

✓ Earle and Perez method
• # of asexual parasites/ 5µl
• thick film is used
• used only in research studies
Thin Blood Film:
✓ % of parasitemia: P. falciparum
✓ # of infected red cells in 1000 RBCs
✓ smear is scanned carefully, one 'row' at a time
✓ total number of red cells and the number of parasitized red cells are tabulated
separately
✓ If 1000 red cells are counted:
• divide the number of parasitized red cells by 10 to get the percentage
• less than 1000 red cells counted
• # of parasitized cells X 100
# of RBCs
"plus system"
✓ less precise
✓ variation in the thickness of the film
✓ results in variation in parasite count
+ = 1–10 per 100 thick fields
++ = 11-100 per 100 thick fields
+++ = 1–10 per thick field
++++ = >10 per thick field
C. COLLECTION OF FECAL SAMPLE

Proper collection of fecal sample is essential for the isolation and proper
diagnosis of infection. The following should be considered:
1. Container: sterile, disposable, wide mouth with tight-fitting lid, transparent
(for physical examination of the specimen)

2. Avoid contamination with urine, water and soil
3. Label
4. Handle carefully because it is a potential source of infection
unsuitable samples from patients receiving:
a. Barium
b. Oil
c. Bismuth
d. Kaolin
e. Antibiotics
TYPES of FECAL SPECIMEN
1. Liquid – either diarrheic or saline-purged

2. Soft
3. Formed
PRESERVATION
a) Physical:
1. Room temperature
2. Refrigeration
b) Chemical
1. Polyvinyl Alcohol (PVA) – used for the preservation of stained fecal+
smears; preservation of trophozoites
2. 5-10% formalin
▪ For concentration techniques or direct fecal smear
▪ For cysts, helminthes and larvae
▪ Not sufficient for preparing permanent stained fecal smears
3. Merthiolate-Iodine-Formalin (MIF) or Thimerosal Iodine Formalin (TIF)
▪ Good for all stages
▪ For liquid stools
▪ For bulk feces, the solution is mixed in the proportion of 9.4 ml
MIF and 0.6 ml Lugol’s for a gram of fecal material
4. Schaudinn’s Fixative

▪ For fresh materials and those recovered from intestinal
mucosal linings
▪ For permanent staining
IMPORTANCE OF FECALYSIS
1. To detect the presence of intestinal parasites

2. For the detection of evidence of malfunction of some parts of the GIT, liver
and pancreas
3. For the detection of GIT bleeding
4. For the detection of excessive fats in stool (steatorrhea)
5. Used as a clue in medical and surgical diagnosis.
D. EXAMINATION OF FECAL SAMPLE

1. Physical Examination
❖ Form and Consistency:
Normal: soft to formed
Variations
a) Very soft and watery – seen in diarrhea and administration of saline
cathartic
b) Excessively hard and scybalous – constipation due to lack of mucus
c) Rice water stool – cholera
d) Pea soup stool – early typhoid
e) Flattened or ribbon like – syphilis, spastic colitis and obstruction at the lower
portion of the colon
f) Butter like- fibrocystic disease of the pancreas
g) Gaseous and fermentative – excessive carbohydrate fermentation
❖ Color
Normal: light brown to dark brown due to stercobilin
Variations
a) Yellow- due to administration of santonin and senna
b) Light clay or putty color – seen after ingestion of Barium meals
c) Reddish or bloody- bleeding in the lower GIT; undigested beets and
tomatoes
d) Dark red/chocolate brown- bleeding in the upper GIT
e) Black/tarry- associated with digestion of blood due to bleeding in the
upper GIT
f) Greenish – amoebiasis
❖ Odor
Normal: foul to offensive due to indole, skatole and butyric acid
Abnormal odors:
a) Putrid odor – found in ulcerative and malignant tumor of the lower bowel
b) Sour/rancid odor- indicates gas formation, fermentation of carbohydrate,
unabsorbed fatty acids
c) Extremely foul odor – usually in alkaline stools, putrefaction of undigested
protein.
2. Chemical Examination
Occult blood – blood which is not visible to the naked eye and can only be
detected by chemical means only
Laboratory Examination for Occult Blood:
a) Benzidine test
b) Guaiac;s test
c) Hematest
General Principle: The peroxidase-like activity of blood (due to peroxidase
contained in the heme portion of hemoglobin) decomposes H2O2 to produce
water and oxygen. The liberation of oxygen causes the oxidation of the
colorless chromogen into a colored compound (blue)
Patient Preparation: meat free diet for 3-5 days prior to the test
3. Microscopic Examination
The following structures may be seen microscopically:
a) Trophozoites and cysts of amoeba
b) Helminth eggs and larvae
c) RBC- due to hemorrhagic disorder, ulcers and contamination
d) Macrophages present in bacterial or parasitic infection
e) WBC- indicative of inflammation

f) Fungi
g) Plant cells, pollen grain and spores
h) Epithelial cells
i) Calcium oxalate and triple phosphate crystals
j) Bacteria
k) Plant fibers, root hairs and animal cells ( similar to helminth ova)
l) Charcot-layden crystals
TECHNIQUES
1. DIRECT FECAL SMEAR – simplest and most frequently used

a) 0.85% sodium chloride/NSS- for the observation of the motility of trophozoites
b) D’antonis solution/Lugol’s Iodine – for protozoan cysts and helminth ova
2. KATO THICK SMEAR (KTS)
▪ Reagents:
Distilled water 100 ml
Glycerin 100 ml
3% malachite green 1 ml
▪ Use cellophane as cover slip
3. CONCENTRATION TECHNIQUES- used in the detection of a small number of
parasites which are not detected using DFS
a) Sedimentation Method – uses either specific gravity or centrifugation
• Formalin-Ether techniques – concentrates helminth eggs, larvae
and protozoan cysts
• Acid-Ether Sedimentation – used for formed stools; applicable for
helminth eggs and larvae
• Simple sedimentation – time-consuming
• Merthiolate-Iodine-Formalin Concentration
• Acid Sodium sulfate Tritan
• Knott Concentration Technique- for concentration of
microfilariae’ utilizes venous blood as specimen
b) Flotation method

✓ Allows the separation of helminth eggs, protozoan cysts and
larvae (1.05 – 1.15) using chemical solutions of higher specific
gravity (1.12-1.21)
✓ Eggs and cysts float to the surface of the solution while fecal
materials sink to the bottom
✓ Superior to sedimentation for concentrating cysts and eggs other
than operculated, schistosomal and infertile Ascaris eggs
• Zinc Sulfate centrifugal flotation technique
- valuable method for concentrating cysts and eggs
- specific gravity: 1.18 (hydrometer); 1:20
formalinized feces
• Sugar Flotation Technique/Sheather’s Sugar Flotation
- Cryptosporidium (rounded oocysts)
• Wilie’s Brine- uses sodium chloride solution
• Lane’s Direct Centrifugal Technique
• Magnesium Sulfate Techniques
4. STAINING METHODS
a) Temporary Staining
• D’Antonis
• 1% Isotonic Eosin Solution & 2% Brilliant Cresyl Blue
- Trophozoites- blue green
- Background- pink
• Eosine in saline
• Buffered Methylene Blue (ex. Nairs Methylene Blue)
b) Permanent Staining
• Gomori’s Trichrome Stain: The trichrome technique of Wheatley for
fecal specimens is a modification of Gomori's original staining
procedure for tissue. It is a rapid, simple procedure which produces
uniformly well stained smears of the intestinal protozoa, human cells,
yeast cells, and artifact material in about 45 min or less.
- Wheatley’s modification

- For intestinal protozoa
• Lawless’ Permanent Mount Stain: A rapid permanent-mount stain
technic for the diagnosis of the intestinal protozoa
- Rapid method of staining trophozoites and cysts
- Protozoa – blue to purplish color
• Modified Acid-Fast stain
- Cryptosporidium, Isospora and Cyclospora
- Two separate stains: Carbol fuchsin and methylene blue
- lacks specificity
- Use of the modified Ziehl-Neelsen stain for faecal smears has
already been established for coccidian protozoa, in particular,
oocysts of Cryptosporidium species, but it is also useful to confirm
the presence of oocysts of Isospora belli and Cyclospora
cayetanensis
• Iron Hematoxylin Stain
- used for most of the original morphological descriptions of
intestinal protozoa found in humans.
- On oil immersion power (1,000 x ), one can examine the
diagnostic features used to identify the protozoan parasite. Iron
hematoxylin staining methods can be used with either fresh, SAF-
preserved, or PVA-preserved specimens.
• MIF Fixative stain ( Merthiolate iodine formaldehyde)
- diagnosis of Trichomonas
• Chlorazol Black E
- An acid dye, used as a fat and general tissue stain, and to stain
protozoa in fecal smears or in tissues.
• Modified Kohn’s
- modification of the chlorazol black E staining technique
5. SPECIAL RECOVERY METHODS
a) Cellulose Tape Preparation/Graham Scotch Tape Method
✓ Enterobius vermicularis and Taenia
✓ Collection: morning before the patient washes or defecates

✓ Commercial collection kits are available
b) Egg Count Technique – provides an estimate of worm burden
✓ Determines the degree of infection (light, moderate or heavy)
✓ For recovery of hookworms, Ascaris and Trichuris
✓ Methods:
1. Direct Smear Egg Count (Method by Beaver)
- 1.5 mg feces (667) = eggs per gram (epg)
- 2.0 mg feces (500) = epg
2. Dilution Egg Count – uses 0.1 N NaOH
- Stoll’s Egg Counting Technique
3. Thick Smear Egg Count – KTS for schistosomes
4. Dilution-Filtration Egg Count – Schistosomes
c) Egg Hatching Technique
✓ Egg hatch assay (EHA) is a laboratory tool used to determine a given
parasite's resistance to extant drug therapy.
✓ Fresh eggs are incubated from the parasite of interest and serial
dilutions of the drug of interest are applied. The percentage of eggs
that hatch or die is determined at each concentration and a drug
response curve may be plotted. The data can then be transformed
and analysed to give further statistics such as an ED50.
✓ This technique is labour intensive, expensive and can take some time,
however an egg hatch assay will give and accurate and reliable
result.
6. CULTURE METHODS
PURPOSES:
✓ For accurate detection of the organisms as a supplement to other
methods
✓ For obtaining a rich yield of organisms to be used as antigen in
immunologic diagnosis
✓ For in-vitro screening of drugs
✓ For investigating the physiology of organisms
✓ As a source for inoculating susceptible experimental animals
CULTURE METHODS for PROTOZOA

a) Balamuth’s Monophasic Medium
- All liquid medium commonly used for the isolation and maintenance
of Entamoeba histolytica and some other intestinal protozoa.
- It is satisfactory for Balantidium coli, although the quality and amount
of starch may need to be adjusted
- Use of egg yolk
b) Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
- Egg slant
- E. histolytica
- Concentrate from saline centrifugal sedimentation of feces rather
than the feces is recommended.
- Use of whole hen’s eggs
c) Cleveland & Collier’s
- This medium is highly specific for E. histolytica , which grows luxuriantly
on this medium. Other intestinal amoebae do not grow readily on this
medium. The technique of overlaying the medium with fresh sterile
horse serum-saline mixture, as reported by Cleveland and Collier, was
reported to be the best method of isolation of E. histolytica .
- Proteose peptone and infusion from liver provide amino acids and
other nitrogenous substances that support growth of E. histolytica .
- Sodium chloride maintains the osmotic balance of the medium and
sodium a-glycerophosphate acts as a phosphorous source.
d) Trussel and Johnson’s medium
✓ Isolation of Trichomonas vaginalis
e) Axenic Culture Method
- histolytica
- Axenic cultivation has been an essential component of numerous
immunological and biochemical studies
- Monophasic clear liquid for rapid growth of dense populations of
amebae
CULTURE METHOD for LEISHMANIA & TRYPANOSOMES
a) Novy, MacNeal and Nicole (NNN) Diphasic

- Best medium

- Use of sterile defibrinated rabbit blood
b) Cellular Media
- For Trypanosoma cruzi and Toxoplasma
IMMUNOLOGIC or SEROLOGIC TESTS – involves antigen-antibody reactions
a) Indirect Hemagglutination (IHA)

- PRINCIPLE: Patient serum and commercial sheep blood cells coated
with the appropriate antigen are required. The serum and cells are
combined and allowed to react. The test sample is then examined
for the presence of red blood cell agglutination
- USES: amoebiasis, Chaga’s disease, malaria, toxoplasmosis,
cystisercosis, hydatid cyst, filariasis, schistosomiasis
b) Flocculation or Agglutination
- PRINCIPLE: The patient serum is added to known cultured parasites.
This mixture is observed for the presence of parasite clumping.
- USES: Leishmaniasis, Chaga’s disease
c) Indirect Fluorescent Antibody (IFA)
- PRINCIPLE: Microscopically visible parasites fixed on a slide are
allowed to react with test serum. The slide is washed and treated with
anti-human globulin with fluorescein (sandwich method)
d) Gel Diffusion (Ouchterlony) and Countercurent Electrophoresis (CEP)
- PRINCIPLE: Patient serum and known antigens are placed in adjacent
wells located on the agar gel. The antigen and antibody are allowed
to migrate across the agar towards one anther. Specific patterns of
precipitate ( precipitin bands) result between the inoculated wells
and are then interpreted.
- USES: Detect amebiasis and fascioliasis infections
e) Enzyme -linked immunosorbent assay (ELISA)
- PRINCIPLE: Test serum is allowed to react with an antigen (coat the
surface of plastic well). Anti-human IgG antibody conjugated with
enzyme is added. If specific antibody was present in the test serum,
the anti-human IgG antibody labeled with an enzyme attaches to

the antigen-antibody complex. The addition of a suitable substrate
generates a visible color reaction.
- USES: T. gondii, T. vaginalis, leishmaniasis, malaria, schistosomiasis etc.
❖ Antibody Capture or Double Sandwich ELISA
✓ Modification of the standard ELISA for increased sensitivity and
specificity in detecting antibodies of the IgM class to
Toxoplasma
✓ Plastic well are coated with anti-human IgM antibodies
✓ After washing, the test serum is added followed by further
washing and addition of the test antigen
✓ The final reagent is an antigen-specific antibody to which the
color reagent has been conjugated
✓ The plates are read spectrophotometrically
✓ This method can be used for assay of parasite-specific IgE
antibodies by starting with anti-human IgE
f) Sabin-Feldman Dye Test
- The Sabin-Feldman Dye Test is based on the fact that live Toxoplasma
tachyzoites can actively take up methylene blue dye from the
culture medium, whereas parasites that are killed because of
complement-mediated lysis do not take up the dye and remain
colorless.
- Specifically designed for the diagnosis of Toxoplasmosis
g) Circumoval Precipitin Test (COPT)
- The circumoval precipitin test is a serological test used for diagnosis of
schistosomiasis japonica. Soluble egg antigens of Schistosoma
japonicum block the formation of the circumoval precipitin by serum
from infected humans. Consequently, circumoval precipitin inhibition
was used to monitor purification of the soluble egg antigens of S.
japonicum.
h) Radioimmunoassay (RIA)

- PRINCIPLE: Radioimmunoassay (RIA) is a very sensitive in vitro assay
technique used to measure concentrations of antigens by use of
antibodies. As such, it can be seen as the inverse of a radiobinding
assay, which quantifies an antibody by use of corresponding
antigens
i) Radioallergosorbent Test (RAST)
- The RAST is a radioimmunoassay test to detect specific IgE antibodies
to suspected or known allergens for the purpose of guiding a
diagnosis about allergy
- In this test, the parasite antigen is first bound to an inert sorbent or
matrix to which the test serum is allowed to react. After washing,
radio-labeled antibody (IgE or IgG) is added to the sorbent. After
another washing to remove excess labeled antibody, the remaining
radioactivity is a measure of antigen-specific IgG or IgE in the test
serum
j) Immunoblotting/Western Blot
- PRINCIPLE: CSF and serum may be used. By using electrophoresis
technique, the DNA containing portion of the patient sample is
transferred to a polyacrylamide gel. This process results in the
separation of protein antigens in the gel. A blotting technique is
employed to transport the proteins onto a paper.
- USES: giardiasis
k) Complement Fixation (CF)
- PRINCIPLE: Three ingredients is necessary to for performing the first
phase: patient serum, a commercial antigen source, known amount
of complement. The complement is “fixed” by the antibodies present
in the serum. In the 2nd phase, sensitized sheep’s rbc is added to the
mixture. The absence of free complement prevents lysis of these cells
( positive). If the patient sera does not contain Ab, complement is
free to lyse ( negative)
- USES: leishmaniasis, Chaga’s disease, pneumocystosis, paragonimiasis
7. ANIMAL INOCULATION TESTS

a) Leishmania
✓ Animal Used: hamster
✓ Specimen: aspirates from biopsy materials from cutaneous ulcers,
lymph nodes, spleen, liver
b) Trypanosoma
✓ Animal Used: Guinea pigs or white rats (Trypanosoma gambiense
and T. rhodesiense)
✓ White mice (Trypanosoma cruzi)
8. XENODIAGNOSIS
✓ An experimental bug (vector) is allowed to take a blood meal from the
patient
✓ The intestinal contents of the bug is assessed for the presence of
diagnostic stages of the parasite
SYNTHESIS
Over the years, parasites which were once considered commensal have evolved to
become human pathogens. During this time, tremendous knowledge was obtained of the
epidemiology, parasite-host relationships, life cycles, disease processes and symptoms,
treatment, and prevention and control of parasites. In addition, parasites are classified
based on their individual characteristics. Traditional as well as new methodologies for
parasite identification allow for accurate laboratory diagnosis.
Parasitology is an interesting and exciting field of the clinical laboratory sciences. The
continued development of high-tech, highly sensitive parasite test methodologies provides
the key to the future of parasitology. Because it is highly unlikely that parasites will totally be
eradicated in the near future, competent practitioners educated in the field of parasitology
are essential to ensure proper parasite identification.
ASSESSMENT
I. RESEARCH ACTIVITY

✓ In not more than 5 sentences, describe and indicate the purpose of the new
techniques in parasitology (20 points). Indicate the reference used. Use your own
words to explain.
1. DNA HYBRIDIZATION PROBE TESTING (DNA PROBE)

2. DNA RESTRICTION FRAGMENT LENGTH POLYMORPHISMS
3. FLOW CYTOMETRY
4. POLYMERASE CHAIN REACTION (PCR)
RUBRIC FOR GRADING:
II. QUIZ
✓ For offline and weak connectivity, it will be provided by the instructor in essay form
✓ For Online/Strong connectivity, it will be provided either through a worksheet, or quizziz
or google forms.
REFERENCES:
Belizario, V. Jr. (1998). Philippine Textbook of Medical Parasitology (1st edition). The
Publications Program.
Zeibig, A. (2013). Clinical Parasitology: A Practical Approach. (2nd ed.). Saunders Elsevier Inc.
Our Wright’s Stain (n.d). Dalyn biologicals (n.d)Retrieved from https://www. dalynn.com
/dyn/ck _assets/ files/ tech/SW80.pdf on July 3, 2020

Leishman staining- Principle, Preparation, Procedure & Result Interpretation (2018).
Paramedics World. Retrieved from https://paramedicsworld.com/hematology-
practicals/leishman-staining-principle-procedure-interpretation/medical-paramedical-
studynotes on July 3, 2020
CLINICAL PARASITOLOGY LABORATORY
LABORATORY GUIDELINES
❖ In case of any future face-to-face classroom/laboratory instructions. Please follow the
guidelines below
STUDENTS
1. Be in your complete white uniform (with nameplate and school ID).

2. Wear your laboratory gowns at all times while inside the laboratory.
3. Incomplete/No uniform /No Laboratory gown = No laboratory performance grade.
4. Put all cellular phones in the silent mode. Answering of cell phones will be dealt
with accordingly.
5. Wash hands thoroughly with soap and water before leaving the laboratory.
6. Long hair should be clipped, fingers and hands should be kept away from the face
and mouth.
ROOM MANAGEMENT & PROPER WASTE DISPPOSAL
1. Strictly no eating inside the laboratory.

2. Maintain cleanliness and orderliness in the laboratory. Refrain from loitering outside
the laboratory room.
3. Disinfect working tables with chlorox, lysol or alcohol before leaving the laboratory.
4. Treat specimens with disinfectants before disposal. Never flush specimens in the
sink. Use the waste cans in disposing remnants.
5. Make use of the waste cans properly. Strictly separate degradable from non-
degradable and infectious from non-infectious wastes.
6. Used slides should be washed or disposed properly. Never leave unwashed slides
on the working tables or in the sink.
7. Return stools under the working tables and pick up pieces of paper before leaving
working area.
❖ The instructions on laboratory reports will be followed regardless of online or face-to-face

instructions
INDIVIDUAL or GROUP REPORTS and QUOTA REQUIREMENTS

1. Whenever a written report is required, references must always be included (Book
title; Author; Edition; Page number) in each item /number of the written report.
Written reports without references will NOT be considered.
2. Online references should contain the complete website address.
3. Record data and label drawings in pen, draw/sketch using pencil (plain or
colored).
4. Submit reports and drawings on time. Late reports and drawings shall be given a
50% deduction from the total score. Reports/drawings submitted on another day
shall be considered for completion purposes only.
5. Margination for written reports are: 1.0 inches – Left; 0.5 inch – top, right & bottom
(in RED ink)
6. Quota specimens should be provided by the students. (Students may obtain
pathologic quota specimens from other clinical laboratories). (as applicable)
7. Slides and cover slips should also be provided by the students. Reagents will be
provided by the central supply office. (as applicable)
PARASITOLOGY LABORATORY DRAWINGS RUBRIC
❖ The rubrics will be utilized for all laboratory activities integrated in this module EXCEPT for
experiments 3 and 4
CATEGORY 4 3 2 1
Labels Every item that Almost all items Most items (75- Less than 75% of
needs to be (90%) that need 89%) that need to the items that
identified has a to be identified be identified have need to be
label. It is very clear have labels. It is labels. It is somewhat identified have
which label goes clear which label clear labels. It is not
with which goes with which which label goes clear which label
structure. structure. with which goes with which
structure. item.
Drawing All assigned details Almost all Almost all assigned Fewer than 85% of
have been added. assigned details (at least the assigned
The details are very details (at least 85%) have been details are
clear and easy to 85%) have been added. A few present.
identify. added. The details are not clear Most details are
details and are difficult not clear and
are clear and to identify are difficult to
easy identify.
to identify.
Accuracy The assigned The assigned The assigned structures The assigned
structures are drawn structures are are drawn with 50-85% structures are
very accurately. drawn with 85- accuracy. drawn with less
and are 95% accuracy. The assigned structures than 50%
recognizable. All All are labelled with 50- accuracy.
assigned assigned 84% accuracy. The assigned
structures are structures are structures are
labeled very labeled with 85- labelled with less
accurately. 95% accuracy. than 50%
accuracy.
Attention to There are no There are 30% or There are 30-50% There are 51%
Detail differences between less obvious obvious differences and more major
the drawing and the differences between the drawing differences
specimen structures. between the and the specimen between the
Work shows attention drawing and the structures. Work shows specimen and
to detail with specimen errors that are the drawing, and
magnification views structures. Work represented when overall details for
and annotation shows minimal comparing the drawing and
information. attention to detail specimen with the information are
with drawing. missing.
magnification
views and
annotation
information..
Formatting All the format 1-2 of the format 3-4 of the format The title, name,
(50%) requirements are requirements are requirements are date, labels and
- Title, Name, present. Magnification missing. missing. Magnification magnification of
Date, Labels, work is done at the Magnification work is shown on back the drawing are
Magnification back of the drawing work is shown at of drawing with less missing.
with more than 75% the back of than 50% accuracy. Magnification
accuracy. drawing with 50- work is not shown
75% accuracy. on back of
drawing.
PARASITOLOGY LABORATORY QUESTIONS FOR RESEARCH RUBRIC
CATEGORY 4 3 2 1
Ideas and All the information 50-80% of the Less than 50% of There is no clear or
Content needed to answer information the information specific explanation in
the question are needed to answer needed to answer to the
present. the question is answer the question.
Supporting details present. The question is
are included. The directions were present. The
directions were followed. directions were
followed. not followed.
Use of terms 76-100% of the 50-75% of the Less than 50% of No terms from the
terms from the terms from the terms from the lesson are used.
lesson are used. lesson are used. lesson are used.
More than 95% of 75-95% of the Less than 75% of
the terms are terms are the terms are
accurately used accurately used. accurately used.
and are fully
defined.
Sentence Fluency More than 75% of At least 75% of the At least 50% of Sentences are
the sentences are sentences are the sentences incomplete or too
complete and complete and are complete long.
clearly understood. clearly and easy to
Connections with understood. understand.
each other are
clearly established.
Conventions Less than 10% have 10-30% have end 31-50% have end More than 50% have
end marks or marks or capital marks or capital end marks or capital
capital letters and letters and letters and letters and structural
structural and structural and structural and and spelling errors.
spelling errors. spelling errors. spelling errors. Answer not clear.
Answer is very Answer is clear. Answer
clear. somewhat clear.

Documentation Uses appropriate Uses appropriate Includes Demonstrates poor
and Referencing evidence and references from a references from a documentation skills
documentation number of few relevant or effort and an over-
demonstrating relevant secondary reliance on tertiary
highly competent information information sources. Silent about
information sources, showing sources, more or unaware of issues
access, evaluation adequate than one tertiary of source accuracy,
and application. information source. reliability, and
Thoroughly access, evaluation Demonstrates authority. Referencing
evaluates source and application. limited lacks consistency of
accuracy, Notes briefly issues bibliographic style and/or format.
reliability, and of source research. Displays
authority. Cited accuracy, inconsistencies
works/bibliography reliability, and about issues of
follow a standard authority. Cited source accuracy,
style and works reliability, and
bibliographic /bibliography authority. Some
format and are follow a standard information is not
skillfully style and referenced.
incorporated into bibliographic Contains several
paper as format with few citation errors.
appropriate citation errors
NAME: RATING:
Laboratory Schedule: Group No:
Date performed: Date submitted:
Experiment No. 1
INSTRUMENTS AND GLASSWARES USED IN CLINICAL PARASITOLOGY
I. Background
The microscope is an essential tool in the study of microorganisms, cells and tissues.
As an instrument, it must be handled and used carefully at all times. In order for the
observer to view organisms under it, it is but necessary that one must know how to use a
microscope effectively.
Aside from the compound microscope, glasswares and instruments are also needed
in Parasitology Laboratory. Glassware is used to contain reagents as well as parasites for
analytical identification. Other instruments are used to facilitate the diagnosis and
identification of parasites.

II. What are expected of you?
At the end of the experiment, you are expected:
1. To describe the different glasswares and different instruments commonly used in

Parasitology
2. To demonstrate the proper operation of the different instruments and glasswares
commonly used in Parasitology
III. What are the materials?
✓ Books
✓ Online resources/references
IV. How do you go about this experiment?

Research on instruments and glass wares. Study the parts, operating techniques and
their uses.
1. Draw and label:

a. Binocular microscope
b. Clinical Centrifuge
c. Glass slides and coverslip
d. Centrifuge tube
e. Plain test tube
f. Beaker
BINOCULAR MICROSCOPE CLINICAL CENTRIFUGE

CENTRIFUGE TUBE PLAIN TEST TUBE
GLASS SLIDE AND COVERSLIP BEAKER
Questions for enrichment (answer on the space provided)
1. What are the quality control measures for the following:
A. Binocular microscope
B. Clinical Centrifuge
C. Glass slides and coverslip
D. Centrifuge tube
E. Plain test tube
F. Beaker

2. What are the different parts of the binocular microscope? List and indicate the
functions of each.
3. Describe the different glass wares/instruments below and give the function of
each in clinical parasitology.
NAME: RATING:
Experiment No. 2
PHYSIOLOGY OF STOOL FORMATION AND PROPER COLLECTION OF SPECIMEN
I. Background
Feces may be defined as the residual mass of material remaining in the intestinal
tract after the full and complete exercise of the digestive and absorptive functions and
are ultimately expelled from the body through the rectum. The semi-fluid intestinal
contents at the duocecal valve are transformed into feces in the large intestine where
the residues remain for one or more days.
1. To enumerate the different organs in the human digestive system

2. To describe the different organs involved in stool formation
3. To demonstrate the proper collection of stool.

Reference materials
Illustration materials/Chart of the Human Digestive System
A. Formation of Feces:
Food is received through the mouth, when food has been chewed to particles
of proper size, mixed with salivary amylase and mucus and shaped into a lubricated
“bolus”; it is now ready for swallowing. The bolus is forced backward through the
mouth and goes down into the pharynx and into the esophagus. Food is forced into
the stomach through the esophagus by muscular movements known as “peristalsis”.
Gentle peristaltic movements pass over the stomach are emptied into the small
intestine. During the passage of intestinal contents through the small intestine,
products of digestion together with many other compounds like vitamins, water,
mineral salts are absorbed. When the contents reach the large intestine, the process
of absorption, with the exemption of water, is normally completed. Here more water
and sodium chloride are absorbed and the remaining material leaves the body as
feces.
B. Proper Collection
1. Stool Container
Stool containers should be covered, clean, wide-mouthed and preferably

colorless. It should be free from water, urine and soil.
2. Time
Submit the specimen immediately to the laboratory and see to it that it is

completely and properly labeled.
Stool examination falls under clinical microscopy in a hospital set-up and thus
may not be given priority since there are other specimens to be examined. However, in
the diagnosis of amoebiasis, consistency of stools dictates that diarrheic or watery stools
must be examined within 30 minutes to 1 hour since trophozoites die within this period
outside the body of the host.
If transportation and examination is to be delayed it should be immediately
preserved in 10%formalin solution or polyvinyl alcohol (PVA).
3. Amount
The amount .of stools to be submitted will depend on the technique that will be
performed. For routine stool examination, 20-40 g of formed stools (usually half of the
thumb) or 5 -6 tbsps of watery stools will suffice.
However, there are situations when it is necessary to submit the whole stool
movement like in cases where the laboratory likes to recover helminthic adults after
treatment. This is best exemplified by the examination done after Taenia species
treatment.
V. Draw and label:

1. Human Digestive System
2. Flow chart of stool formation
HUMAN DIGESTIVE SYSTEM FLOW CHART OF STOOL FORMATION
Questions for enrichment (write answers on the space provided)

1. What are the importance of proper collection of fecal sample?
2. What is the importance of delivering the stool specimen in the laboratory as soon
as possible?
3. Give the reasons why stool should not be mixed with toilet water and urine.
4. List different IMPROPER collection of fecal sample
NAME: RATING:
Experiment No. 3
ROUTINE STOOL EXAMINATION: PHYSICAL & CHEMICAL PHASE
I. Background
A normal stool consists chiefly of undigested food particles, various products of
digestion and a large amount of bacteria, which are usually non-pathogenic. When
diseases occur, food particles increase since the food material is swept out before
complete digestion or absorption can take place. Many of these food remnants can
be identified either macroscopically, such as seeds, fruit or vegetable skins, etc., or
microscopically, such as undigested muscle fibers, vegetable cells, vegetable fibers
etc.

1. To differentiate the normal and abnormal physical characteristics of stool

2. To be able to perform routine chemical examinations on fecal samples.
3. To describe and interpret the results of chemical tests performed on fecal
samples

✓ Books

Procedure:
A. Physical Examination: take note of the .color, odor and Consistency of the fecal
sample.
Normal: Color - yellow or brown
Odor: offensive but excessively
Consistency: soft to semi-formed
B. Chemical examination:
Occult Blood - Benzidine method
1. Smear a pea size stool on a strip of filter paper

2. Put drops of saturated acetic acid with benzidine powder at the center of your
smear.
3. Add an equal amount of 3% hydrogen peroxide.
4. Read the result not after than 2 minutes after the addition of the last reagent.
Results and Interpretations:
Trace - very faint blue
(+) - Faint blue
(++) - Distinct blue
(+++) - Dark blue
(++++) - prussian blue
V. Results and observations:

Color:___________________________ Odor:_____________________
Consistency:______________________ Benzidine test:______________
VI. Draw and label:

1. Positive and neqative results/ Standard Results of the benzidine
POSITIVE STANDARD RESULT NEGATIVE STANDARD RESULT

1. In a table form, list down the different colors of stool with their corresponding
clinical significance
STOOL COLOR ASSOCIATED DISEASES
2. In a table form, list down the different stool consistency with their
corresponding clinical significance
STOOL ASSOCIATED DISEASES
CONSISTENCY
3. Explain the importance of the occult blood test. Illustrate through a flow chart
the principle of occult blood.
4. List the causes of false positive and false negative results in occult blood test.
5. What is the patient preparation prior to occult blood test?
6. What is the test to distinguish between the presence of fetal blood or maternal
blood in an infant’s stool or vomitus? Discuss the principle and the expected
results.

NAME: RATING:
Experiment No. 4
ROUTINE STOOL EXAMINATION: MICROSCOPIC
I. Background
The microscopist should be familiar with the different structures found in the feces
such as trophozoites and cysts of Amoeba, helminth eggs and larvae, RBC,
macrophages, WBC, fungi, plant cells (pollen grains and spores), epithelial cells, crystals
like Calcium oxalate and triple phosphate, bacteria, plant fibers, root hairs and animal
cells are similar to helminth ova.

Most common errors in the identification of parasites may be due to the following:
(1) Lack of familiarity with the parasites, (2) Cursory examination of the slide and (3)
Confusion of parasite with artifacts.

To be familiar with the different method and techniques commonly employed in

the identification of parasites using the stool sample

Slides and Cover slips 10% formalin solution
Applicator sticks ether or ethyl acetate
NSS gauze
Lugol's solution Centrifuge tubes
KTS (Cellophane strips soaked in malachite green solution of glycerol)
✓ Books

Procedure:
A. Direct Fecal Smear (DFS) or Direct Wet Mount Technique
This is the simplest and moderately efficient procedure for examinations of feces. The
mixing action of the intestinal tract usually results in an even distribution of parasites in stool.
DFS preparations from soft, loose and watery specimens should be mandatory. It is highly
recommended for the detection of motile trophozoites in liquid, diarrheic and bloody
mucoid specimens, however, if the number of organisms are few, this method may be of
low sensitivity and insufficient to reveal their presence.
1. Comminute approximately 2 mg of feces with a drop of NSS on a clean slide

using an applicator stick until a uniform suspension without fibers or gritty
materials obtained.
2. Place the cover slip above the preparation and immediately examine under
the LPO of the microscope in a systematic method (figure A). Confirm
identification under HPO.
3. Wet mounts may be preserved for several hours by ringing the cover glass with
paraffin or nail polish.
Figure A: method for systematically scanning a wet mount preparation.
Notes:

The amount of feces used for the direct mount is important. The 2 mg recommended
approximates what would form a low cone at the end of a wooded applicator stick.
When less than 2 mg of feces is used, the suspension will be too thin and may have
blank spaces, whereas the use of more than 2 mg results in a suspension that is too thick
and parasites may be hidden under fecal debris.
Temporary stains may be used with wet mount preparations to aid in location and
identification of protozoa but are not necessary for eggs and larvae. Dilute iodine solutions
are the most commonly used temporary stains and are most useful for recognition of cyst
stage; however, they kill and distort the trophozoites. In cysts, the visibility of nuclei is
enhanced so that their number and morphological features are more clearly seen.
The use of weak iodine solutions is not recommended since they do not stain organisms
well. Likewise, the use of an iodine solution that is too strong will stain the organisms so
darkly that morphological features cannot be seen. Dilute iodine solutions that are
recommended are D' Antoni's, Dobell and O'Connor's and Lugol's.
B. Kato - Thick Smear Preparation:

This technique is based on the fact that when a layer of stool is in contact with glycerin-
soaked cellophane for a period of time, it becomes clarified and helminth eggs become
visible. .
1. Place a pea-size stool sample at the center of glass slide and cover with a square
piece of pre-treated cellophane.
2. With the aid of a rubber stopper, press the cellophane gently to spread the stool,
taking care that the specimen does not spread beyond the cellophane cover,
the cellophane also serves as a cover slip.
3. Leave the prepared slide at room temperature for 10-20 minutes. During this time
the microscopic field becomes clear due to the action of glycerin on the stool
constituents.
4. The slide should be examined after 10-20 minutes or within one hour after
preparation. Allowing the slide to stand for long period of time will cause drying
and shells of hookworm ova will become be difficult to see.
C. Formalin - Ether Concentration Technique (FECT)

The use of concentration procedures for fecal examination virtually ensures the
detection of even small numbers of organisms that would go undetected if only direct
smears of permanent-stained slides were examined.
Concentration procedures generally fall into two categories - flotation and

sedimentation. Sedimentation can be achieved by simple gravity or centrifugation. Since
the sediment will generally contain all the parasites occurring in the stool sample, this
procedure has greater diagnostic sensitivity. An added advantage is that the technique
can be readily applied to both fresh and preserved feces.
1. Thoroughly comminute approximately 1.0 to 1.5 g of fresh feces in 10 ml 10% formalin

in a suitable container. Let it stand for 30 minutes or longer to achieve adequate
fixation. For very loose or watery fecal sample, use 5-6 ml of material.

2. Strain the suspension through two layers of wet gauze and pour into a 15 ml conical
centrifuge tube.
3. Fill tube with NSS; centrifuge at 400 - 500 g for 1 – 2 minutes.
4. Discard the supernatant if cloudy; the sediment should be resuspended then
centrifuged again using NSS. Proceed to the next step only after the supernatant is
clear.
5. Resuspend the sediment in 10% formalin to a volume of 10 ml; add 3 ml of ether (or
ethyl acetate) then shake vigorously for 30 seconds. If ether is used, hold tube with
stopper directed away from face.
6. Centrifuge at 400 - 500 g for 2 -3 minutes. .When tube is removed, it will be seen to
consist of four layers:
a. a top layer of ether
b. a plug of debris adhering to the wall
c. a layer of formalin
d. sediment for examination. Insert applicator stick with cottoned tip to ring and
loosen the plug of debris
7. Decant tube, discard top 3 layers, mix small amount of fluid left with the remaining
sediment and pipette out.
8. Prepare iodine and unstained wet mounts for examination
Notes:
Recently, concern over storage and use of ether, a potentially flammable and
explosive material, has led to the use of ethyl acetate as a substitute. Ethyl acetate
appears to be somewhat more efficient than ether when specimens contain Taenia or
Hymenolepis nana eggs or Giardia cysts since there are fewer tendencies for these eggs
or cysts to become trapped in the plug of debris.
However, one disadvantage is that ethyl acetate, when used as a solvent for
fresh feces, is not as efficient as ether in extraction of fatty or mucoid materials which
may interfere with the microscopic examination of the sample
V. Draw and label:

a. Print/ Copy paste and Label:
1. a picture of a DFS stool smear under the microscope
2. a picture of a KTS stool smear under the microscope
b. Draw and Label:
1. Procedures A, B and C
PRINT and LABEL (Microscopic)

DFS
KTS
PROCEDURE A
PROCEDURE B
PROCEDURE C

1. List the reagents used in KTS and the importance of each
2. Discuss the purpose of concentration technique
3. Enumerate and describe the concentration methods
4. Enumerate and describe the different flotation methods
5. Identify the specific parasites that can be isolated in each special method

SECTION 2: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF NEMATODES
(ROUNDWORMS)

1. identified each member of Phylum Nematode according to morphology, and

recognize stage of the parasite being studied
2. Valued the importance of ethics of observing patients’ and results’ confidentiality
3. Maintained ethical standard in working with other health personnel
LESSON PROPER
This part begins with the discussion of the helminths. The first group of helminths
discussed are the nematodes, commonly known as the intestinal roundworms.
GENERAL CHARACTERISTICS:
1. General morphology: Unsegmented, elongated and cylindrical in shape. The body is

covered by a non-nucleated cuticle which may be smooth striated, bossed or
ornamented with spines.
• Pseudocoele – body cavity which contains all of the viscera (digestive,
excretory, reproductive and nervous system.
• Anterior end – mouth/buccal cavity which may be provided with spines,
hooks, cutting plates, stylets, or other structures for attachment or penetration
• Length – a few mm to a meter in length
• Movement – by sinuous changes of their bodies
2. The sexes are separate, and the females are larger than the males; male posterior end
is usually curved

3. Reproductive system
• Male: testes, vas deferens, seminal vesicle, ejaculatory duct, cloaca
a) accessory copulatory apparatus
b) gubernaculum (wing- like appendage)
c) telamon
d) copulatory spicule
e) copulatory bursa (umbrella like ex: Hookworms)
4. Digestive System: with complete alimentary tract
• Esophagus: filariform, rhabditiform, spiruroid, strongyliform or stichosoma
5. Nervous System: with chemoreceptors
• Acetyl choline- excitatory neurotransmitter
• Gamma-amino butyric acid (GABA) – inhibitory neurotransmitter
• AMPHIDS: pair of laterally placed minute receptor organs in the cephalic or
cervical region of all nematodes.
• PHASMIDS: pair of minute lateral post-anal organs in species w/o caudal glands
6. Excretory System: with collecting tubules or collecting canal & an excretory pore
7. No circulatory system (fluid of the body cavity contains hemoglobin, glucose, proteins,
salts and vitamins)
8. No respiratory system
9. Developmental stages: 1) egg stage 2) four larval stages 3)) adult stage
10. Reproduction: The adult female may be
a) Oviparous (ex. Ascaris)
b) Viviparous (ex. Trichinella)
c) Parthenogenetic (ex. Strongyloides)
11. Modes of attachment
• Anchorage with attenuated ends (T. trichiura)
• Oral attachment to the mucosa (Ancylostoma)
• Penetration of the tissues (Strongyloides)
• Retention in the folds of the mucosa (lumcricoides)
12. Modes of nourishment
• By sucking with ingestion of blood
• By ingestion of lyzed tissues and blood by embedded worms

• By feeding with intestinal contents
• By deriving from body fluids
13. Life span
o Trichinella spiralis: 4 – 16 weeks
o Enterobius vermicularis: 1 – 2 months
o A. lumbricoides: 12 – 17 months
o Hookworms: at least 14 years
o Filarial worms: up to 25 years
o T. trichiura: 5 – 10 years
o S. stercoralis: 20 – 30 years
14. Life Cycle
o Egg - three layers:
Vitelline membrane
Chorionic or true shell
Albuminous covering
o Four larval stages
o Adult stage
CLASSIFICATION OF MEDICALLY IMPORTANT GROUPS AND SPECIES OF NEMATODES
Class: Aphasmidia (lack “Phasmids” or without caudal chemoreceptors)
A. Species which parasitize the small intestine

1. Trichinella spiralis
2. Capillaria philippinensis
B. Species which parasitize the large intestine
1. Trichuris trichiura
Class: Phasmidia (with “Phasmids”)

1. Ascaris limbricoides
2. Necator americanus
3. Ancylostoma duodenale
4. Ancylostoma ceylanicum

5. Strongyloides stercoralis
B. Specie which parasitize the large intestine
1. Enterobius vermicularis
C. Species which parasitize the tissues
1. Wuchereria bancrofti
2. Brugia malayi
3. Loa loa
4. Dracunculus medinensis
D. Species which causes larva migrans in man

1. Ancylostima braziliense
2. Ancylostoma caninum
3. Toxocara spp.

SECTION 2A: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF APHASMID
NEMATODES (ADENOPHOREA)

LESSON PROPER
The Adenophorea (also called the Aphasmidia) seems to be the most primitive group
of nematodes. Mainly, they are free-living in soil and water; however, there are a few
parasitic forms of aphasmids. As the alternate name implies, they do not have phasmids,
and the amphids are located posteriorly on the head region. In fact, they have no sensory
bristles or papillae on the head and body. They are simple, spindle-shaped worms with
simple excretory organs (single-celled).
1. Trichinella spiralis
• Synonym: Trichinella spiralis
• Common name: Trichina worm
• Infective stage: Encysted larva
• Main Habitat: small intestine, skeletal muscles (larva)
• Final Hosts: hogs, rats,
man
• Intermediate hosts: same
animal as the final host
• Developmental stages:
1. Larva: at birth: 80-120 µm
& highly coiled
: has spear-like burrowing tip
at its tapering anterior end
: grows rapidly about 1 mm
2. Adults: rarely seen in stool
or any material
✓ Females: 3.5 mm x 0.06 mm
: Posterior end: bluntly rounded; anterior fifth: with single
vulva
LARVIPAROUS
✓ Males: 1.5 mm x 0.04 mm
: posterior end: ventrally curved with two lobular caudal
appendages

• Life span: 4-16 weeks
• Infective
stage: ENCYSTED
LARVAE
• Mode of transmission:
ingested of
encysted larva
• Life Cycle: BLIND
ALLEY CYCLE
- requires 2
hosts in order to
complete the life
cycle
• Human infection is a
dead-end infection
• Disease: Trichinosis, Trichiniasis, Trichinelliasis

Stages of pathologic changes:
1. Intestinal: inflammation of duodenal & jejunal mucosa, malaise, nausea,

diarrhea and abdominal cramps
2. Muscle invasion: fever, facial edema (particularly in the eyes), pain, swelling
& weakness of the involved muscle, eosinophilia
3. Convalescence: begins at about the end of the 3rd week; s/s subside; cyst
wall then the larva itself CALCIFY
3 clinical phases
1. Intestinal Phase
✓ first week
✓ small intestinal edema and inflammation
✓ nausea, vomiting, abdominal pain, diarrhea, headache and fever,
myalgia
2. Migration Phase
✓ up to 6th week:
- high fever (40oC)
- blurred vision
- edema of the face and eyes
- cough, pleural pains
- eosinophilia (15-40% for 1 month)
✓ 4th to 8th week: death
3. Muscular Phase
✓ acute local inflammation
✓ edema and pain of the musculature
✓ Larval encystation
✓ muscle fibers
- 3-4 days after larval invasion
- Edematous
- spindle shape
- lose their cross striations
- undergo basophilic degeneration
- Nuclei:
size and number
migrate to the anterior of the
muscle cell
Predominating symptoms
✓ Eosinophilia
✓ orbital edema
✓ muscular pain and tenderness
✓ shallow, painful breathing
✓ general weakness
4. Encystment/Encapsulation Phase
✓ fever, weakness, pain and other symptoms start to abate
• Diagnosis
1. Muscle Biopsy:
✓ gastrocnemius and biceps
✓ digest with pepsin-hydrochloride
✓ 3rd or 4th week of infection
2. Bachman Intradermal Test
✓ use antigen preparation of Trichinella larva
✓ Observe after 30 minutes
✓ (+) result: large elevated swelling at the site of injection
3. Beck’s Xenodiagnosis
4. Serodiagnosis
5. Bentonite Flocculation
6. Blood Count : Eosinophilia
• Prevention:
✓ Elimination of encysted larva in hogs through freezing at -30oC for 24 hours
✓ Extermination of rats and mice around farms
✓ Thorough cooking of pork
2. Capillaria philippinensis
• Common name: Pudoc worm

• Infective stage: 3rd stage larva
• Intermediate host: glassfish, “bagsit”, “bagsang”, “ipon”
• DH: Man and birds
• Main Habitat: L.I. & S.I.
• Distribution: Philippines & Thailand
• Developmental stages
1. OVA
✓ Color: Pale yellow

✓ Size: 42x20 µ
✓ similar to that of T. trichiura
✓ smaller & more striated shells
✓ Flattened plugs
✓ Peanut shape
2. FEMALE
✓ size: 2.4 – 4.3 mm
✓ Divisions:
✓ Anterior – esophagus and esophageal glands
✓ Posterior – intestine and reproductive organs
❖ Atypical female
- uterus lined with 2-3 rows of eggs
- Larviparous
- causes internal auto-reinfection
❖ Typical female
- uterus lined with 1 row of egg
- oviparous
3. MALE
✓ size: 2.3 – 3.17mm
✓ caudal alae; long, non-spiny sheath
• Disease caused: Capillariasis or Mystery disease
• Predominant Symptoms
✓ borborygmi
✓ abdominal pain
✓ Diarrhea (chronic)
✓ Untreated:
✓ weight loss; malaise
✓ vomiting; dehydration
✓ anorexia
✓ pneumonia, heart failure and cerebral edema
✓ Death: 2 – 8 weeks after these are seen
• Treatment
1. Albendazole
✓ drug of choice
✓ 400 mg/day for 10 days
✓ destroys larvae readily
2. Mebendazole
✓ 200mg twice a day for 20 days or 400 mg/day for 20 days
3. Electrolyte replacement therapy and high protein diet
• Prevention: Thorough cooking of fish

• Laboratory Diagnosis: DFS & Concentration technique
• Mode of Transmission: eating infected fish

3. Capillaria hepatica
• Common name: Capillary liver worm

• Infective stage: embryonated ova
• Hosts: rats, dogs, cats, monkeys and rarely in human
• Habitat: liver of the host
1. Ova:
✓ “lemon – shaped” eggs
✓ resembles that of Trichuris trichiura
✓ outer shell: pitted like golf ball
2. Adults
✓ males are half as long as the female with slightly chitinized spicule
• Diagnosis: Liver biopsy (revealing characteristic egg and parasite)
• Transmission:
✓ Eating of dog’s meat
✓ Ingestion of contaminated food and drinks (embryonated ova)
4. Trichuris trichiura
• Synonyms: Trichocephalus trichiurus, Trichocephalus dispar
• Common name: whipworm
• Principal host: man, but has been found in hogs, monkeys, cattle, dogs,
mice
• Main habitat: cecum & appendix
• Life span: 5-10 years
1. Ova:
✓ Barrel/football-shaped, Japanese lantern
✓ 50 µmX25µm in size
✓ 3 layers:
✓ undeveloped, unicellular embryo
✓ outermost layer – smooth, bile-stained
✓ Hyaline/Mucus plug
2. Adult
✓ flesh – colored
✓ anterior three-fifths is attenuated (whiplike)
✓ stichosoma type of esophagus
a) male
- 30-45 mm
- posterior portion: coiled ( 360o)
- lanceolate spicule protruding through a refractile
pineal sheath
b) Female:
- 35-50 mm
- bluntly rounded posterior end
- 3,000-10,000 eggs per day
• Disease:
✓ Trichuriasis

✓ Trichucephaliasis
✓ Whipworm infection
- Slight infection – asymptomatic
- Heavy infection – surface of colon matted with worms
• Pathology & Symptomatology
✓ Asymptomatic for light infections
✓ Heavy infection: surface colon is matted with worms
- Bloody or mucoid diarrhea
- Weight loss and weakness
- Abdominal pain and tenderness
- Increased peristalsis and rectal prolapse
- Obstruction and inflammation of the appendix (appendicitis)
- Hypoalbuminemia and IDA
- Extreme cachexia
• Diagnosis: DFS, KTS, Concentration techniques
• Treatment: Mebendazole (500mg) , Albendazole (400 mg) & Oxantel-pyrantel
• Prevention
- Sanitary disposal of feces
- Thorough washing of hands
- Thorough washing and cooking of food
- Avoid using human feces as fertilizer
5. Dioctophyma renale
• Synonym: Eustrongylus gigas

• Common name; Giant Kidney Worm
• Infective stage: 3rd stage larva
• Habitat: Kidney (typically the right kidney)
• Hosts:
✓ Definitive - Mink, dogs, foxes, and other carnivores; fish eating mammals
✓ Intermediate - Oligochaete annelids
✓ Paratenic - Fish and frogs.
• Life span: 5 years
• Diagnosis: eggs in urine (Urine sedimentation)
• Treatment: surgical excision
1. Ova:
✓ ellipsoidal and brownish-yellow
✓ deeply sculptured depressions
2. Adults:
✓ blood red in color
✓ attenuated (slightly) on both ends
a) Males:
- Size: 14-20 cm x 4-6 mm
- Bell-shaped copulatory bursa
- not supported by rays; covering of papillae
b) Females
- vulva: midventral near anterior
SYNTHESIS
Aphasmids are nematodes which lack phasmids or it lacks caudal chemoreceptors.

The nematodes in this classification are:
✓ Trichinella spiralis

✓ Capillaria philippinensis
✓ Capillaria hepatica
✓ Trichuris trichiura
✓ Dioctophyma renale
ASSESSMENT
QUIZ
or google forms.
REFERENCES:
Aguinaldo, A. M. A., J. M. Turbeville, L. S. Linford, M. C. Rivera, J. R. Garey, R. A. Raff, and J. A.

Lake. 1997. Evidence for a clade of nematodes, arthropods and other moulting
animals. Nature. 387:489-493.
Diesing (n.d). Description of the Phylum Nematoda. Retrieved from

https://comenius.susqu.edu/biol/202/animals/protostomes/ecdysozoa/nematoda/nem
atoda-description.html on July 10, 2020

LABORATORY
NAME: RATING:
Experiment No. 5
PHYLUM NEMATODA - CLASS ENOPLEA (ADENOPHOREA; APHASMIDEA)
I. Background
Papillae, amphids and phasmids are the main sensory organs of nematodes. Most sensory organs are
found in the anterior end, but males often have well-developed sensory structures associated with
copulation. Phasmids are paired structures found posteriorly. The presence or absence of phasmids had
been used to classify nematodes into two classes.
In class Adenophorea; Aphasmidea, the amphids are generally well developed (except in parasitic
forms), well behind the lips that are often pocket like. Caudal and hypodermal glands are common.
Phasmids are absent. The excretory system lacks lateral canal and are formed of single, ventral,
glandular cells, or entirely absent. Dierids are absent. Most of the members are free living and some are
parasitic on plants and animals.
General characteristics of the parasitic species within the Enoplea:
A. with 5 esophageal glands

B. generally with caudal and hypodermal glands
C. phasmids absent
D. excretory system formed of single, ventral, glandular cells or may be entirely absent
1. To know the morphology, life cycle, and laboratory methods employed in the identification of
Nematodes.
2. To be familiar with the diagnostic and infective stages of these nematodes.
3. Identify the nematodes using the distinguishing marks of their diagnostic stages
4. to know the mode of transmission of these nematodes

Colored plates
Reference Book
a. Print and label: Microscopic (HPO)
1. ovum of Trichuris trichiura
2. ovum of Capillaria philippinensis
3. encysted larvae of Trichinella spiralis
b. Draw and label: Diagrammatic
1. Adult male and female (typical and atypical) Capillaria philippinensis
c. Label the parts: Diagrammatic
1. Adult male and female Trichuris trichiura
2. Adult male and female Trichinella spiralis
PRINT and LABEL ( Microscopic)
ovum of Trichuris trichiura ovum of Capillaria philippinensis
encysted larvae of Trichinella spiralis
DRAW and LABEL (diagrammatic)
Adult male (typical and atypical) Capillaria philippinensis

Adult female (typical and atypical) Capillaria philippinensis
LABEL THE FOLLOWING:
1. ADULT MALE and FEMALE Trichuris trichiura

( http://mt-
lectures.blogspot.com/2016/07/lecture-2-nematodes.html)
A. ADULT FEMALE B. ADULT MALE

1. _____________________ 1. _____________________
2. _____________________ 2. _____________________
3. _____________________ 3. _____________________
4. _____________________ 4. _____________________
5. _____________________ 5. _____________________
6. _____________________ 6. _____________________
7. _____________________ 7. _____________________
8. _____________________ 8. _____________________
9. _____________________

2. ADULT MALE and FEMALE Trichinella spiralis
(https://www.pinterest.ph/pin/558164947550122566/)
MALE
1. ______________________________
2. ______________________________
FEMALE
1. _____________________________
2. _____________________________
3. _____________________________
4. _____________________________
5. _____________________________
6. _____________________________
7. _____________________________
8. _____________________________
9. _____________________________

1. In a table form, differentiate Trichuris trichiura, Capillaria philippinensis and Trichinella spiralis based
on: ova, morphology, diagnostic stage, infective stage, other name, disease caused, distinguishing
mark or characteristic, mode of transmission
2. Describe the life cycle of Trichuris trichiura, Capillaria philippinensis and Trichinella spiralis
3. Give and describe the laboratory methods in the identification of Nematodes.

SECTION 2B: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF PHASMID
NEMATODES (PHASMIDEA)

LESSON PROPER
The Secernentea (also called the (phasmidia) vary greatly in size from microscopic to
several feet long. The largest known secernentean, which is up to 30 ft (9 m) in length, lives in
the placentas of female sperm whales. The body of secernenteans consists of a flexible
cylinder that tapers at both ends, with a pointed tail and a blunt head. They are considered
non-segmented pseudocoelomates; that is, creatures possessing a three-layered body that
has a fluid-filled body cavity (pseudocoelom) between the endoderm and the mesoderm
(the innermost and middle tissue layers).
A flexible but tough collagenous cuticle surrounds the body with a system of grooves
across the body from head to tail, which protects them internally. The non-cellular cuticle
varies from four to two layers and is almost always transversely striated. Laterally for most of
the body length, the cuticle is generally modified into a wing area that is marked by
longitudinal ridges; generally, this region is only slightly above the normal body contour.
However, in some parasitic forms, it may extend out a distance equal to the body's diameter.
The cellular hypodermis is the subcuticular layer that secretes the cuticle.
The sensory system contains phasmids, which are a pair of bilateral cuticular,
glandular organs, situated laterally in the caudal (posterior to the anus) region and opening
to the surface by a slit or pore. Also known as precaudal glands, phasmids are unique to the
secernenteans, in which their function is believed to be sensory. At the other end are pore-
like amphid apertures, which are a pair of glandular chemosensory organs situated dorso-
laterally in the cephalic (head or anterior) region and opening through the cuticle. Although
usually pore-like, in isolated instances the aperture can be an oval or a cleft. The apertures
show little variation throughout the secernenteans. The amphids are always labial (located
on the lips). The external amphidial aperture is usually less well developed than in
adenophorean worms.
Somatic and cephalic setae, which are elongated structures jointed with the cuticle,
are rare. When present, the cephalic sensilla are located on the labial region, and they are
pore-like or papilliform. In males, there may be caudal setae. In females, somatic setae are
absent. Generally, sixteen sensilla are present in the shape of two circles (an inner circle of
six, and an outer circle of 10). In some parasitic groups, the number of cephalic sensilla may
be reduced. Deirids, pairs of pore-like sensilla that usually protrude above the surface of the
cuticle, are usually present on the cervical region near the level of the nerve ring.
Secernenteans contain no hypodermal glands, but the hypodermal cells of the

hypodermis (a thin tissue layer beneath the cuticle that thickens to form the dorsal, ventral,
and two lateral hypodermal chords, and extends the length of the body) are usually
multinucleate (syncytial: more than two nuclei), but may also be uninucleate (cellular: one
nuclei). These divide the muscle cells into four fields. A layer of longitudinal muscles underlies
the hypodermis.
Bursae, or caudal alae, are sublateral projections generated by a longitudinal

widening of the cuticle. It is common for them to be present within male secernenteans,
each looking like a fluid-filled body sac. The thin cuticle extensions are located on both sides
of the anus, specifically on either side of (or sometimes surrounding) the cloaca (the
urogenital opening) of males. The well-developed excretory system is in the shape of an H or
U. It is a simple tubular system that is located in one or both of the lateral hypodermal chords,
and embedded between the three cell bodies in the hypodermal chord. The system opens
ventromedially by way of a cuticularized duct. The rectal part of the gland system is usually
present. They have no caudal glands.
The muscular esophagus or pharynx varies in configuration, but the majority of

secernenteans have three esophageal glands: two that are subventral and one that is
dorsal. The subventral glands open into the posterior metacarpus. The dorsal gland opens

anteriorly into the procorpus or the anterior metacarpus. Its basic structure is corpus (the
anterior part is cylindrical), with the basal (bottom) region sometimes swollen in the shape of
a bulb. The glands empty their contents into the esophagus to aid in digestion. The tail is the
region between the anus and the back tip of the body.
1. Ascaris lumbricoides
• Most common intestinal roundworm

• Common name: Giant Intestinal roundworm
• Definitive host: man (no intermediate host needed)
• Main habitat: Lumen of the SI
• Life span: 12-17 months
1. Ova/Eggs
Types of eggs
a) Unfertilized: longer and narrower
2 layers of the egg shell
✓ albuminoid layer (absent in old specimens)
✓ chorionic layer or true shell
➢ filled with amorphous mass
➢ lack the cresentric clear area
b) fertilized: broadly avoidal and thick
3 layers:
✓ Chorionic/true shell: chitinous layer; secretory product of the
egg
✓ Vitelline layer: fertilization membrane; highly impermeable
membrane that protects the inner embryo
✓ Protein coat/Albuminous layer: outermost mamillated layer with
a tanning action
➢ Embryonated: same as fertilized but contains the larva of the
embryo
➢ Decorticated: lacks the albuminous mamillated shell; usually seen
in old specimens; it may be fertilized or unfertilized
2. Adult: white, creamy or pinkish yellow when freshly expelled and resembles
earthworm (lumbricus); head is provided with three conspicuous lips
which are finely denticulated; each lip has minute twinned sensory
papillae.

a) Male
➢ 10-31 cm
➢ Usually shorter and slender
➢ ventrically curved posterior end with 2 spicules
➢ genitalia: composed of a single, long tortuous tubule
b) Female
➢ 35 cm long x 3-6 mm
➢ straight posterior end
➢ paired reproductive organs located in the 2/3 of the body
➢ oviparous
➢ gravid uterus : 200,000 eggs
• Disease: Ascariasis, Dooryard or Backyard Infection
• Pathology
a) Due to larval migration:
✓ Ascaris pneumonitis: Damage to the pulmonary tissue
(petechial hemorrhage) when larvae break out of the lung
capillaries into the air sacs
✓ Symptoms manifested: asthmatic type of respiration; cough;
bronchial rales (abnormal respiratory sound); urticarial rash
(hives, vascular reaction of the upper dermis; eosinophilia in the
circulatory blood)
b) Due to adult worms:
✓ Diarrhea, vague abdominal pain, nausea and loss of appetite
✓ Due to its erratic behavior: vomiting; suffocation; intestinal
obstruction, appendicitis; acute pancreatitis; peritonitis
(perforation of the bowel)
• Diagnosis: DFS, KTS, Concentration Technique, ELISA
• Stool examination may give negative results due to the following
- During the early stage of infection (worms are still immature)
- During larval migration through the blood stream
- When only male worms are present in the intestines
• Prevention:
✓ Sanitary disposal of human excreta
✓ Personal hygiene
✓ Avoid the use of night soil fertilizer
✓ Thorough cooking of food particularly vegetables and washing of fruits
✓ Washing solution: aqueous iodine solution (200 parts/million)
✓ kills infective egg and larva in 15 minutes
• Treatment: Mebendazole (500mg), Pyrantel pamoate (10 mg/kg (maximum
of 1 g)), Albendazole (400 mg)

2. HOOKWORMS
Human Hookworms:
a) Necator americanus: American hookworm? American murderer?New World

Hookworm
b) Ancylostoma duodenale: Old world hookworm
c) Ancylostoma ceylanicum
Animal Hookworms: (cause skin lesions or larval migrans in man)
d) Ancylostoma braziliense (cat hookworm)

e) Ancylostoma caninum (dog hookworm)
1. Ova: ovoidal, colorless/hyaline and thin shelled; 56-60 µ x 34-40µ
: 4-8 cell stage when passed in the feces (surrounded by a clear zone)

2. Rhabditiform larva:
➢ Feeding stage, short and stout
➢ Has a long narrow buccal cavity
➢ Flask shaped esophagus
➢ Very small genital primordium
3. Filariform larva:
➢ Non-feeding
➢ Infective
stage
➢ Mouth is
close with a
protecting
sheath
➢ Longer and
slender with a
pointed
posterior end
4. Adult hookworms:
➢ small, cylindrical, fusiform, grayish white
➢ relatively stout with a cervical curvature which appeared like a hook
➢ with a well-developed buccal capsule
Females: 9-13 mm by 0.35 to 0.6 mm
Males: 5 – 11 mm by 0,3-0,45 mm
HOOKWORM SPECIE BUCCAL CAVITY COPULATORY BURSA

Necator americanus - 1 pair of semilunar cutting - fused spicules; deep cleft
plates, dorsal median - bipartite dorsal rays
tooth, deep pair of
triangular subventral
lancets
Ancylostoma duodenale - 2 pairs of fused ventral - unfused spicules; shallow
teeth cleft
- tripartite dorsal rays
Ancylostoma caninum - 3 pairs of ventral teeth - bursa is supported by
long, slender rays
Ancylostoma braziliense - 1 pair of larger outer - bursa is supported by
teeth and 1 pair of very short stubby rays
inconspicuous median
teeth
• Disease: Necatoriasis/Uncinariasis: Necator americanus
Ancylostomiasis: Ancylostoma species
• Pathology:
a) Due to larval stage:
1. Ground itch/Coolie itch/Dew itch
✓ Dermatitis at the site of entrance of filariform

✓ Intense itching, edema and erythema and later
papulovesicular eruption
2. Creeping eruption/Cutaneous larval migrans/Plumber’s itch/
Duck Hunter’s itch
✓ Due to the exposure of the skin to the filariform larvae of
A. braziliense and A caninum, occasionally of N.
americanus and A duodenale.
✓ “Serpiginous tunnel” in the stratum germinativum of the
skin
3. Pumonary lesions: Wakana disease
✓ Petechial hemorrhages with eosinophilic and leukocytic
infiltration that induces cough and pyrexia
b) Due to adult worm
1. Hookworm anemia
✓ Chronic blood loss due to continuous mechanical
suction of blood from the intestinal mucosa and the
presence of bleeding areas left by the adult as they
transfer to new areas
✓ Blood loss: N. americanus: 0.03-0.05 ml/day
A duodenale: 0.16 – 0.34 ml/day
✓ Blood picture: “Mycrocytic Hypochromic Anemia”

2. Hypoalbuminemia
✓ Loss of protein due to a combined loss of blood and
lymph and the protein loss is as well in excess of the loss
of RBC
• Diagnosis:
a) Ground itch and creeping eruption – characteristic of the lesion
and the history of skin contact with soil
b) Recovery of eggs – DFS, KTS, Brine flotation and FECT
c) Harada Mori culture technique
• Treatment
a) Mebendazole, Pyrantel pamoate, Oxantel

b) Severe anemia- raise the hemoglobin level to about 70-80 g/l. Iron
therapy (Ferrous sulfate, 200 mg 3x a day for 3 months)
• Prevention:
a) Sanitary disposal of feces
b) Avoid sites where infected dogs and cats may defecate
c) Eradicating the infection in dogs and cats by periodic anti-
helminthic treatment
d) Personal hygiene such as use of shoes or slippers
e) Avoiding ingestion of raw vegetables

3.
Strongyloides stercoralis
• Common name: Threadworm

• Definitive host: man
• Habitat: Upper small intestines (duodenum)
• 2 species: S. stercoralis, S. fuellerborni
1. Ova
➢ ovoidal thin shelled, transparent, resembles a Chinese lantern
➢ not found in feces except in diarrhea and hyperistalsis
➢ contains a fully developed embryo
2. Rhabditiform larvae
➢ Flask-shaped & stout esophagus

➢ Short buccal cavity
➢ Conspicuous genital primordium
3. Filariform larvae
➢ Non-feeding stage with a long and delicate esophagus
➢ Forked or notched tail
4. Adult
➢ well – developed buccal capsule
➢ no teeth, no cutting plates
➢ but bears a crown of chitinous, leaf-like processes
2 PHASES of development
1. Parasitic
✓ inhabits the intestine of host

✓ female is a delicate filiform worm
✓ L: 2.2 mm
✓ esophagus: occupies 1/3 of the anterior part (longer)
✓ Parthenogenetic
2. Free-living
✓ Exists in the environment

✓ buccal cavity: slightly larger that that of the parasitic male
worm
✓ adult female: shorter, smaller
✓ shorter esophagus
Transmission: skin penetration; autoinfection
• Disease: Cochin-china diarrhea; Strongyloidiasis, Strongyloidosis

• Pathology and Symptoms:
1. Skin: allergic, raised red blotches at the site of larval penetration
2. Migration of larvae: bronchial verminous pneumonia
3. Intestine: abdominal pain, diarrhea and constipation, vomiting, weight
loss, variable anemia, eosinophilia, protein losing enteropathy
❖ Asymptomatic in light infection
4. Death in immuno-compromised patients due to heavy autoinfection or

larval migration throughout the body
• Diagnosis: DFS, stool culture (Harada mori filter paper tech) rhabditiform larvae
in feces; Enterotest; Baermann, ELISA
• Treatment: Thiabendazole – ovicidal & larvicidal
Albendazole – 400 mg/day for 3 days
❖ Strongyloidiasis is difficult to treat
❖ Internal infection can continue for years because of
autoinfection

• Prevention: same as hookworms
4.
Enterobius vermicularis

• Synonym: Oxyuris vermicularis
• Common name: pin worm, seat worm
• Main habitat: cecum and appendix
• Reservoir host: dogs and cats
• Infective stage: embryonated egg
1. Ova
➢ Double lined chorionic shell, transparent and colorless; elongated
and ovoidal with one side flattened
➢ With inner embryonated layer and outer albuminous shell
➢ Embryonated when laid at the perianal area
➢ Remain viable up to 13 days, rarely seen in the stool
2. Adult
➢ Small, spindle-shaped, relatively stout with dorsoventral bladder-like
expansions of cuticle called the “cephalic alae”/”lateral wings”
➢ Have an oral end and three lips, hour glass-shaped esophagus
a) Male: 2-5 mm long, strongly curved pointed tail which is used for
copulation, spicule is conspicuous
b) Female: 8-13 mm in length by 0.4 mm; posterior end is sharply
pointed; vulva found in the middle third; paired genital organs
• Disease: Enterobiasis, Oxyuriasis
• Mode of Transmission
✓ By anus to mouth via contaminated fingers and fomites
✓ Through contaminated food and drinks especially if the food handler is the
carrier
✓ Via inhalation- viable ova can float in the air
✓ Retro infection: gravid female after laying their eggs in the perianal area
goes back through the anus to the large intestine. The larvae upon
hatching migrate back to the large intestine
• Pathology & Symptomatology:
✓ Some are asymptomatic; rarely causes serious lesions

✓ Other symptoms
- Nocturnal perianal itching
- Vulva irritation; vulvovaginitis, salpingitis
- Cardinal feature: hypersensitivity
- Mild nausea or vomiting
- Loss of sleep, irritability
- Slight irritation to intestinal mucosa
• Control & Prevention: extremely difficult once infection sets in the household
✓ Home and community sanitation
✓ Better personal hygiene; fingernails should be cut short
✓ Use showers rather than bath tubs
✓ Infected persons should sleep alone
• Diagnosis
a) Graham Scotch Tape Technique/Cellulose Acetate Technique
b) NIH Swab
Technique
c) Schuffner and
Swelling Rebel
Method

TOXOCARA SPECIES:
• Toxocara canis (Dog Ascarid)

• Toxocara cati ( Cat Ascarid)
➢ Synonyms: Ascaris mystax/Belascaris mystax/Belascaris cati
• Can infect humans and cause damage of the visceral organs
• Morphology: adult female
➢ L: 10-12 cm
➢ passes numerous eggs into their host’s feces
• Disease
There are two major forms of toxocariasis:
1) Ocular larva migrans (OLM):
✓ Toxocara infections can cause OLM, an eye disease that can cause blindness.
2) Visceral larva migrans (VLM):
✓ Heavier, or repeated Toxocara infections, while rare, can cause VLM, a disease
that causes swelling of the body’s organs or central nervous system.
✓ Liver lesions: gray, elevated, circumscribed, diameter: 4 mm
✓ granulomatous lesions: eosinophils, lymphocytes, epitheloid cells and giant cells
of foreign-body type
✓ Larvae: liver, brain, eye, spinal cord, lungs, cardiac muscle, kidney and lymph
nodes
• Symptoms: due to the inflammatory reaction at the site of infection
➢ Benign : 20-80% eosinophilia and hepatomegaly
➢ Severe: Intermittent pain, dermatitis and neurologic disturbances, Pneumonitis.
Liver and spleen enlargement, Skin rashes on the lower extremities
➢ Larvae: liver, brain , eye (blindness-most serious), spinal cord, lungs, cardiac
muscle, kidney and lymph nodes

• Treatment: Mebendazole to eliminate the worm
: Prednisone for inflammatory symptoms
• Prevention: avoidance of infected dogs and cats
• Diagnosis
➢ Clinical s/s: triad of marked eosinophilia, hepatomegaly & hyperglobulinemia
➢ History of exposure to dogs, cats and dirt eating
➢ ELISA; EIA
GNATHOSTOMA SPECIE
• 1st IH: copepods

• 2nd IH: fish and amphibians

• Diagnosis: migratory lesions, eating raw fish
• LENGTH: 2 to 3 centimeters long and are rust-colored
• LARVA:
➢ four rows of hooklets extruding from the surface of the cephalic bulb
➢ Tiny, cuticular spines run along the length of their bodies
➢ Two types of papillae extend from the worm--a cervical papilla off the main
body and two labial papillae on the cephalic bulb
➢ Four sac-like openings in the cephalic bulb
• EGGS/OVA:
➢ ovular with a mucus plug at one end
➢ approximately 40 micrometers to 70 micrometers
• may cause a VLM like syndrome (Southeast Asia). The larvae may migrate through
subcutaneous tissues, causing transient swelling, and to deeper tissues, eventually
invading the CNS
• Gnathostomiasis (Gnathostoma spinigerum)
➢ food-borne parasitic infection that results from the human ingestion of the third-
stage larvae
➢ OTHER TERMS:
- Choko-Fushu Tua chid or chokofishi (Japan),
- consular disease (Nanjing),
- Shanghai rheumatism,
- Tau-cheed (Thailand),
- Woodbury bug (Australia), and
- Yangtze River edema
➢ MOT:
- eating undercooked or raw freshwater fish, eels, frogs, birds, and reptiles
- contaminated water
- In rare instances, larvae can directly penetrate the skin of individuals who
are exposed to contaminated food sources or freshwater.
➢ SIGNS & SYMPTOMS:
- migratory swellings under the skin
- increased levels of eosinophils in the blood.

- Rarely, the parasite can enter other tissues such as the liver, and the eye,
- vision loss or blindness,
- It can also affect the nerves, spinal cord, or brain, resulting in nerve pain,
paralysis, coma and death.
TRICHOSTRONGYLUS SPP
• Zoonotic infection (herbivores)

• Human infection: T. colubriformis, T. orientalis, T. axei, T. brevi
Trichostrongylus tenuis: Infects poultry and other birds worldwide
Trichostrongylus axei.:The stomach hairworm. Infects cattle, sheep, goats, pigs and
horses and many wild mammals. Found worldwide
Trichostrongylus colubriformis. Bankrupt worms, black scours worms: Infects cattle,
sheep, goats, pigs and horses and many wild mammals. Found worldwide
Other less frequent species are Trichostrongylus probolurus and Trichostrongylus vitrinu
• Habitat: small intestines: eggs – larvae-ingestion by DF
• Disease: light & asymptomatic; heavy infection may produce abdominal pain and
diarrhea, usually with eosinophilia
Trichostrongylosis or Trichostrongyliasis
- worms damage the lining of the small intestine or the stomach
- enteritis, gastritis, and sometimes anemia
- Typical signs are diarrhea (mucous and/or hemorrhagic) or constipation
- general weakness and wasting
- loss of appetite
- reduced weight gains or even weight loss, etc.
- Acute severe infections in young animals may be fatal
• Eggs resemble those of hookworms (78-98 µm by 40-50 µm)- slightly tapered at one
end
• Diagnosis:
➢ detection of characteristic eggs in the feces.
➢ determination of the species requires post mortem examination of adult worms
after necropsy
• Life cycle
➢ Direct life cycle
➢ Adult females lay eggs in the large intestine of the host that are shed with the
feces. Once in the environment the eggs release the L1-larvae that complete
development to infective L3-larvae in about 5 days by suitable weather (hot
and humid), significantly longer by cold weather. These infective larvae can
survive in the environment and remain infective for up to 6 months.
Angiostrongylus cantonensis
• Synonym: Pulmonema cantonensis

• Common name: rat/rodent lungworm
• Definitive host: rats Main habitat: lungs
• Disease: Human Angiostrongylosis
• Mode of transmission: ingestion of larvae in raw or undercooked snails or other vectors,
or contaminated water and vegetables
• Infective stage: 3dr stage larva
1. Ova
➢ Elongated, ovoidal with a delicate hyaline shell: 46-48 to 74 microns
2. 1st stage larva
➢ Has a distinct dorsal minute notch near the tip of the tail
3. 3rd stage larva
➢ With 2 well developed chitinous rods with expanded knob-like tips at the
anterior end
4. Adult worms
➢ Filiform worms with a length of 17-25 mm
a) FEMALE: the milky white uterine tubules are spirally wound around the blood
filled intestine and can be seen through transparent cuticle as a “barbers
pole” pattern
b) MALE: has a well developed caudal bursa which is kidney-shaped and
single lobed
• Pathology & Symptomatology
➢ Confusion, incoherence, disorientation and impairment of memory of
profound coma
• Diagnosis:
➢ Brain dyscrasia- moderate to high eosinophilic counts in the spinal fluid
➢ History of the patient as to ingestion of snails, crabs or leafy vegetables
➢ Autopsy to make thorough gross and microscopic examination of the brain
and spinal cord
• Transmission
➢ Ingestion of raw mollusks containing the 3rd stage larva
➢ Ingestion of raw leafy vegetables contaminated with mucus secretions of the
mollusks containing the 3rd stage larva
➢ Drinking water contaminated with the infected larva
➢ Ingestion of paratenic hosts such as fresh water prawns and crabs containing
the infective larva
• Prevention & Control
➢ Thorough cooking of prawns and crabs
➢ Thorough washing of leafy vegetables
➢ Avoidance of ingestion of raw Achatina fulica
➢ Elimination of the snail and eradication of rodents

Anisakis species
• nematode of planctonic crustacean, sea fish and sea mammals

• Common gastrointestinal parasites of marine mammals (with Pseudoterranova)
• Disease: Anisakis:Anisakiasis or Herring Disease
➢ Anisakis spp appear to be more prone to produce invasive disease
➢ Pseudoterranova spp tend to be coughed up or vomited intact
➢ Eustrongyloides spp have been reported in individuals who had eaten live
minnows or home prepared sushi. These parasites usually infect fish-eating birds,
but in humans , the bright red larvae invade the abdominal cavity, requiring
surgical removal.
• Mode of Transmission: ingestion of 3rd stage larva in the flesh of raw fish

• Anisakis Diagnosis: confirmed by the recovery of an intact worm at endoscopy or by
the presence of an eosinophilic granuloma containing an identifiable nematode in a
surgical specimen
• Prevention:
➢ Freezing the fish -20oC for 24 hours
➢ Fishermen are required to eviscerate the fish soon after the catch
• Treatment:
➢ Depending on the severity of the infection, it might require medical attention.
➢ The larva(e) can be removed surgically or during endoscopic examination.
➢ Albendazole
Dracunculus medinensis
• Common name: Guinea

worm; fiery serpent of the Israelites;
Medina worm, serpent worm or dragon worm
• Disease: Dracunculiasis /Dracontiasis/Dracunculosis

• Habitat: cutaneous and subcutaneous tissue
• Life span: 12-18 months
• Morphology: female measures 50-120 cm long by 0.7 -1.7 mm in diameter
Male measures 12-29 by 0.4 mm
• Pathogenesis & Symptomatology
➢ If worms fail to reach the skin, it dies and either disintegrates or calcifies
➢ Mesenteric tissues- it causes pseudoperitoneal syndromes and allergic
manifestations.
➢ Superficial tissues- the worm liberates a toxic substance that produces local
inflammatory reactions; blisters appear at any location and rupture of blister
favors the escape of larva.
➢ Contamination of ruptured blister causes secondary bacterial infection such as
abscesses, cellulitis, extensive ulceration and necrosis
• Diagnosis
➢ Visual observation of painful skin blister- outline of worm under the skin is
revealed by reflected light; emerging worm
➢ Larvae release may be induced when cold water is applied
➢ Calcified worm may be located by x-ray
• Treatment
➢ Mebendazole, niridazole, thiabendazole
➢ Surgical removal of the worm or slow withdrawal from blister by gentle traction
and rolling the protruded portion on a stick (few cm per day)
➢ Aspirin for pain and prevent secondary infection
• Prevention & Control
➢ Infected individuals must not be allowed to bathe or wade in water used for
drinking purposes
➢ Refrain from drinking from suspected water source
➢ Boiling of water. Provision of piped water or wells

BLOOD AND TISSUE NEMATODES IN MAN
• Filarial worms
• Arthropod transmitted/mosquito-borne parasites
• Infective stage to man: filariform/filiform/3rd stage microfilariae
• Infective stage to vector: microfilariae
• Diagnostic stages: Microfilariae: blood
Adult larvae: lymphatics
• Morphology:
1. Adult- threadlike, creamy, white, varies in length (2-50 cm)
2. larval stage- snake-like with a column of cells in the anterior to the posterior portion
3. Microfilaria-pre-larval stage, embryos produced by filarial worms usually found in the
blood or tissues of patients with filariasis, highly motile and threadlike
❖ Location of Microfilariae in man: peripheral blood and lymph spaces of the skin
• Sheath: a thin, translucent egg shell remnant covering the body of the microfilaria and
past the head and tail
SHEATHED Microfilariae
2. Brugia malayi
3. Loa loa
UNSHEATHED Microfilariae
1. Onchocerca volvulus
2. Dipetalonema perstans/Mansonella perstans
3. Dipetalonema streptocerca/Mansonella streptocerca
4. Mansonella ozzardi
• Periodicity: rhythmical appearance of microfilariae in the peripheral blood

Types:
1. Periodic:
➢ Nocturnal – 10pm to 2am
➢ Diurnal – 10am to 2pm
2. Subperiodic:
➢ Nocturnally – peak count during night time
➢ Diurnally – peak count during day time
3. Non-periodic: during night time and day time
• Disease caused: Lymphatic Filariasis

• Clinical Manifestation:
a) Hydrocele: swelling in the scrotum that occurs when fluid collects in the thin sheath
surrounding a testicle.
b) Lymphedema: swelling that generally occurs in one of the arms or legs
c) Elephantiasis: condition characterized by gross enlargement of an area of the
body, especially the limbs
d) Chyluria: rare condition in which lymphatic fluid leaks into the kidneys and turns the
urine milky white
• Laboratory Diagnosis for Filariae
1. Microscopic Examination
a) Wet smears & thick smears – microfilariae
➢ Stains: Wrights; Giemsa; Delafield Hematoxylin
➢ Negative results: low intensity of infection; dead worms; obstructed
lymphatics
➢ DEC (Diethylcarbamazine) provocative tests (3mg/kg single dose)-
stimulates microfilariae to come out to the peripheral circulation
b) KNOTT’S method
➢ 1 ml WB + 10 ml 2% formalin
➢ Centrifuge at 500 x for 10 minutes
➢ Sediment- thick and thin smears: Microfilariae & WBCs
➢ Centrifugation of the blood sample lyzed in 2% formalin (Knott’s
technique)
c) Filtration: Nucleopore or Millipore membrane (5 µm pore)
➢ 1 ml of fresh or anticoagulated blood is drawn up into a syring
➢ Lyze by adding 10 ml distilled water then pass through the Swinney filter
membrane
➢ Examine filter membrane
d) Skin snips: microfilariae of O.volvulus and M. streptocerca
➢ Use corneal-scleral punch, or a scalpel and needle
➢ Incubate sample for 30 minutes to 2 hours in saline or culture medium
then examine for microfilariae that would have migrated from the tissue to
the liquid phase of the specimen
2. Capillary tube method
➢ Use heparinized capillary tube; examine buffy coat layer
3. Immunoassay
➢ Antigen detection- circulating filarial antigens

➢ Molecular diagnosis using PCR- differentiation of filarial species and
stage
4. Identification of adult worms

➢ Ultrasonography- demonstrate live worms in the lymphatics
➢ Contrast lymphangiography & lymphscintigraphy using radiolabeled
albumin or dextran
➢ Tissue samples collected during nodulectomies (onchocerciasis)
➢ Subcutaneous biopsies or worm removal from the eye (loiasis)
• Treatment
1. DEC (Diethylcarbamazine)/Hatrazan
➢ A filaricidal piperazine derivative that kills both microfilariae and some adult
worms of Wuchereria bancrofti and Brugia malayi
➢ Periodic mass treatment: Single dose of DEC for 6 months
➢ 6 mg/kg body weight for 12 days (bancroftian filariasis)
➢ 3 – 6 mg/kg body weight up to a total dose of 36-72 mg/body weight
(brugian filariasis)
▪ Use of DEC fortified tablet salt ( 0.1% DEC or 0.3% DEC for 3-4
month
2. Ivermectin (single dose of 200-400 µg/kbw for 12 days)
▪ More effective if used in combination with DEC
▪ Relief of pain- cool the affected area
• Prevention
➢ Abaca workers-wear long sleeves shirts

➢ Use of mosquito repellants and/or mosquito nets; vector control (insecticides)
SHEATHED FILARIAL WORMS
Wuchereria bancrofti Brugia malayi Loa loa

Common Bancroft’s filarial worm Malayan filarial worm Eye worm, Loa
name worm
Disease(s) Bancroftian filariasis, Malayan filariasis, Loasis, fugitive
caused elephantiasis of lower elephantiasis of upper swelling, calabar
extremeties, extremeties swelling(diameter:5-
lymphatics of scrotum; 10 cm); migration
wuchereriasis rate under the skin:
1 inch/2 mins

Morphology MALE: 4 cm in length MALE: 13-23 mm in length MALE: 30-34 mm by
by 0.1 mm FEMALE: 43-55 mm 0.35 -0.43 mm
FEMALE: viviparous, 8- FEMALE: 40-70 mm
10 cm in length by 0.2- by 0.5 mm
0.3 mm
Development inside Development inside Maturation inside
MOSQUITO: 6-20 days MOSQUITO: 2 weeks MOSQUITO: 10-12
VESSELS OF MAN: 6 VESELS: 3-9 mons days
mons SUBCUTANEOUS
LIFE SPAN: 5 years TISSUES: 1 year
LIFE SPAN: 1-15
years
Habitat Lymph vessels and Lymph vessels and lymph Subcutaneous and
(adult lymph glands (lower) glands (upper) muscular tissues
worm)
Specimen blood blood blood
Vector Culex pipiens Mansonia spp Chrysops spp
quinquefasciatus Anopheles spp (Deer fly /Mango
fly)
Endemicity In the Philippines, the province endemic for W.
bancrofti are: Camarines Norte & Sur, Bohol,
Samar, Albay, Leyte, Sorsogon, all provinces in
Mindanao, Quezon, Mindoro, Mt. Province, Sulu,
Masbate, Palawan, Romblon
W. bancrofti
- Anopheles minismus var, flavirostris – rural type
- Aedes poecillus- urban type of bancroftian
filariasis (both breed in the axils of abaca &
banana)
B. malayi
- Mansonia bonnae (in fresh water)
-Mansonia uniformis (rice fields)

DIFFERENTIAL CHARACTERISTICS OF SHEATHED MICROFILARIAE
Wuchereria Brugia malayi Loa loa

bancrofti
Length 200-300 µm 220-250 µm 250- 300 µm
Diameter 8 µm 6 µm 8 µm
Sheath Stains slightly w/ Stains deeply w/ Almost colorless
Giemsa Giemsa w/Giemsa
Body curves Regular smoothly Irregular and Irregular and
curved twisted twisted same as B.
Graceful sweeping Stiff w/secondary malayi
kinks
Cephalic space Small/short (1:1) Large/longer (1:2) Larger/longer (1:2)
Body nuclei Coarse/well Coarse, tend to Coarse tend to
separated overlap overlap
Tail end No nuclei pointed 2 widely spaced Nuclei present
tip nuclei blunt tip rounded tip
Periodicity nocturnal nocturnal diurnal
UNSHEATHED FILARIAL WORMS
Onchocerca Mansonella Mansonella Mansonella

volvulus perstans streptocerca ozzzardi
Common Blinding worm, Synonyms: Ozzard’s
name convoluted Dipetalonema,Acanthocheilonema
filaria

Disease(s) Onchocerciasis, dipetalonemiasis streptocerciasis Ozzard’s
caused onchocercosis, filariasis,
river blindness mansonelliasis
ozzardi
Morphology MALE: 19-42 cm MALE: 45 mm in FEMALES: 65-81
by 130-210 µm length mm by 0.21-
FEMALE: 50 cm FEMALES: 80 mm 0.25 mm
by 270-400 µm in length
Habitat Subcutaneous Inside body Inside body
tissues cavities cavities,
mesentery and
visceral fat
Specimen Skin snips blood blood blood
Vector Simulium Culicoides spp Culicoides spp Culicoides spp
damnosum (night biting)
(black flies;
buffalo fly)
Endemicity Found in Africa Found in Africa
DIFFERENTIAL CHARACTERISTICS OF UNSHEATHED MICROFILARIAE
Onchocerca Mansonella Mansonella Mansonella

volvulus perstans streptocerca ozzzardi
Length 250-300 µm 150-200 µm 180-240 µm 150-200 µm

Diameter 8 µm 4 µm 5 µm 4 µm
Sheath absent absent absent absent
Body curves Regular slightly Regular often Tail usually Regular slightly
twisted forms loops curved twisted
Cephalic Large and large large large
space bulbous
Body nuclei Coarse mostly Medium sized Fine mostly Fine mostly
separated tend to separated separated
overlap
Tail end No nuclei Nuclei present Nuclei present No nuclei
pointed tip rounded tip curved tip pointed tip
Periodicity Non periodic Non periodic Non periodic Non periodic

SYNTHESIS
Phasmids are unicellular sensilia in the lateral tail region of certain species of
nematodes. They are similar in their structure to amphid sensilla, but smaller. Phasmid neurons
were recently shown to function in modulation of chemorepulsion behavior.. The nematodes
in this classification are:

1. Ascaris limbricoides
2. Necator americanus
3. Ancylostoma duodenale
4. Ancylostoma ceylanicum
5. Strongyloides stercoralis
B. Specie which parasitize the large intestine
1. Enterobius vermicularis
C. Species which parasitize the tissues
2. Brugia malayi
3. Loa loa
D. Species which causes larva migrans in man
1. Ancylostima braziliense
2. Ancylostoma caninum
3. Toxocara spp.
ASSESSMENT
SEATWORK: Complete the table below
DIFFERENCES BETWEEN HOOKWORMS & STRONGYLOIDES

STRONGYLOIDES HOOKWORMS
RHABDITIFORM
Buccal cavity
Genital primordium
FILARIFORM

Esophagus
Tail
QUIZ
or google forms.
REFERENCES
Encyclopedia.com. (2020) Secernentea (Secernenteans) Environment Encyclopedias

almanacs transcripts and maps Secernentea (Secernenteans) Retrieved from
https://www.encyclopedia.com/environment/encyclopedias-almanacs-transcripts-
and-maps/secernentea-secernenteans on July 12, 2020



LABORATORY
NAME: RATING:
Experiment No. 6
PHYLUM NEMATODA - CLASS PHASMIDEA
I. Background
Superfamily Ascaridoidea to which Ascaris lumbricoides belongs have lips that are often prominent.
The buccal capsule is weakly cuticularized and surrounded by esophageal tissue. The esophagus consists
of corpus and posterior ventriculus which may be muscular or glandular. The eggs are thick shelled. The
life cycle is direct or indirect with invertebrate or vertebrate intermediate hosts.
Order oxyurida have medium to small worms often with sharply pointed tails. The esophagus is
characterized with prominent posterior bulb with valve. Eggs are often flattened on one side. Enterobius
vermicularis belongs to this group.

1. To be able to know the morphology of the ova and adult members class phasmidea
2. To describe the life cycle of Ascaris lumbricoides and Enterobius vermicularis
3. To know the mode of transmission for both Ascaris lumbricoides and Enterobius vermicularis
4. To be familiar with the diagnostic and infective stages of these nematodes.
5. Identify the nematodes using the distinguishing marks of their diagnostic stages

Microscope
Prepared slides
Colored plates
Reference Book

1. ova of Ascaris lumbricoides
2. ova of Enterobius vermicularis
b. Label: Diagrammatic

1. decorticated ovum of Ascaris lumbricoides
2. fertilized ovum of Ascaris lumbricoides
3. unfertilized ovum of Ascaris lumbricoides
4. adult male and female of Ascaris lumbricoides
5. Adult male and female of Enterobius vermiclaris
PRINT and LABEL (microscopic)
ova of Ascaris lumbricoides

ova of Enterobius vermicularis
LABEL (diagrammatic)
A. DECORTICATED OVUM of Ascaris

lumbricoides
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwjgtPCDm5bcAhVLn5Q
KHdqiBNIQjxx6BAgBEAI&url=https%3A%2F%2Fwww.slideshare.net%2Fparasitologypera%2Fintestinal-
nematodes&psig=AOvVaw3pevTH4BDHF2aFHB8c9Sra&ust=1531369743890007)

1. _____________________________
2. _____________________________
3. _____________________________
B.FERTILIZED OVUM of Ascaris lumbricoides
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=&url=http%3A%2F%2Fwww.biologydi
scussion.com%2Fparasites%2Fstool-examination-for-parasitic-
infestation%2F29978&psig=AOvVaw3pevTH4BDHF2aFHB8c9Sra&ust=1531369743890007)
1. _______________________________
2. _______________________________
3. _______________________________
4. _______________________________
5. _______________________________
C. UNFERTILIZED OVUM of Ascaris lumbricoides

(http://www.biologydiscussion.com/parasites/stool-examination-for-parasitic-infestation/29978)
1. _____________________________________
2. _____________________________________
3. _____________________________________
D. ADULT MALE and FEMALE of Ascaris lumbricoides

(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=&url=http%3A%2F%2Fwww.biologydisc
ussion.com%2Fplants%2Freproductive-system-and-life-cycle-in-ascaris-with-
diagram%2F2703&psig=AOvVaw346_ifFRttEWLa-UY9Xv0B&ust=1531372413799998)
A. ADULT MALE (A) B. ADULT FEMALE (B)
1. _____________________ 1. _____________________
2. _____________________ 2. _____________________
3. _____________________ 3. _____________________
4. _____________________ 4. _____________________
5. _____________________ 5. _____________________
6. _____________________ 6. _____________________
7. _____________________ 7. _____________________
8. _____________________ 8. _____________________
9. _____________________ 9. _____________________
10. _____________________ 10. _____________________
11. _____________________ 11. _____________________
E. ADULT MALE and FEMALE of Enterobius vermiclaris

(https://www.emaze.com/@ACORTZLW/Enterobius)
A. ADULT MALE (A)

B. ADULT FEMALE (B)
1._________________________
2._________________________ 1. _____________________
3. _________________________ 2. _____________________
4. _________________________ 3. _____________________
5. _________________________ 4. _____________________
6. _________________________ 5. _____________________

1. Describe the morphology of the ova and adult members of class Phasmidea
2. Describe the life cycle of Ascaris lumbricoides and Enterobius vermicularis
3. Differentiate A. lumbricoides and E. vermicularis based on: common name, disease caused, mode of
transmission, diagnostic and infective stage, distinguishing mark/s
4. List and describe laboratory methods to identify and isolate members of class Phasmidea.

NAME: RATING:
Experiment No. 7
PHYLUM NEMATODA - CLASS RHABDITEA (SECERNENTEA; PHASMIDEA)
I. Background
Amphids are paired sensory organs located at the level of the cephalic papillae. They are found in all
nematodes, but are inconspicuous in the terrestrial and parasitic nematodes. Although generally
considered as chemoreceptors, recent studies have shown that they may have secretory functions as well.
In Class Rhabditea, the amphids are ventrally coiled or derived there from. There are three
esophageal glands; the subventral glands open near the base of the esophageal corpus while the dorsal
gland opens at or near buccal cavity.
The amphids in Subclass Rhabditia are generally poorly developed, with small simple pores near or
on the lips; caudal and hypodermal glands are absent. Phasmids are present. The excretory system is with
one or two lateral canals, with or without associated glandular cells. Dierids are commonly present.
Organisms are usually free living .in soil or fresh water or parasitic in plants or animals.
Order Rhabtidita is characterized with tiny to small worms, commonly with six small lips. The
esophagus is muscular and divided into anterior corpus, median isthmus, and posterior bulb. The bulb is
usually present in parasitic species. The tail is conical in both sexes. Most are free living but some are
parasites in the lung of amphibians and reptiles or in the intestine of amphibians, reptiles, birds, and
mammals.
Order: Rabiditida
Families: Rhabdiasidae, Strongyloididae
Genus: Strongyloides
Specie: stercoralis
Members of the Order Strongylida are commonly long, slender worms. The esophagus is usually
swollen posteriorly but lacking a definite bulb. The males are with a well-developed copulatory bursa
supported by sensory rays. The first-, second-, and beginning of third-stage juveniles are free living or
parasitic in invertebrates. They are usually oviparous. The eggs are thin-shelled, rarely developed beyond
morula when laid. They are parasites of all classes of vertebrates.
Order: Strongylida
Genus: Ancylostoma, Necator
Specie: duodenale, americanus

1. To be able to know the morphology of the ova and adult members class Rhabditea
2. To describe the life cycle of Hookworms and Strongyloides stercoralis
3. To know the mode of transmission for both Hookworms and Strongyloides stercoralis
4. To be familiar with the diagnostic and infective stages of Hookworms and Strongyloides
stercoralis.
5. To be able to differentiate morphologically the Hookworms from Strongyloides
6. To know the laboratory methods used for their identification.
Microscope
Prepared slides
Colored plates
Reference Book
a. Print and label : Microscopic (HPO)
1. ova of Hookworm
2. Rhabditiform and Filariform larvae of Hookworm
3. Rhabditiform and filariform larvae of Strongyloides stercoralis
b. Print with proper labels: Diagrammatic
1. Necator americanus - Buccal cavity and copulatory bursa of males
2. Ancylostoma duodenale – Buccal cavity and copulatory bursa of males
3. Ancylostoma caninum - buccal cavity and copulatory bursa of males
4. Ancylostoma braziliense - buccal cavity and copulatory bursa of males
5. Ancylostoma ceylanicum - buccal cavity and copulatory bursa of males
c. Label: Diagrammatic
1. Adult male and female hookworm (A. duodenale)
2. Adult male and Female Strongyloides stercoralis
3. Filariform and Rhabditiform larvae of Hookworms versus Filariform and
Rhabditiform larvae of Strongyloides stercoralis

ova of Hookworm Rhabditiform larvae of Hookworm
Filariform larvae of Hookworm Rhabditiform larvae of Strongyloides stercoralis
filariform larvae of Strongyloides stercoralis

PRINT with PROPER LABEL (diagrammatic)
Specie Buccal cavity Copulatory bursa of males

Necator americanus
Ancylostoma duodenale
Ancylostoma caninum
Ancylostoma braziliense
Ancylostoma ceylanicum

ADULT MALE and FEMALE HOOKWORM ( A. duodenale)

(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwjs2rT9yJjcAhXJGZQKHc40BLQQjxx6BAgBEAI&url=http%3A
%2F%2Fwww.biologydiscussion.com%2Fparasitology%2Fparasitic-worm%2Fstructure-of-adult-hookworm-with-
diagram%2F62228&psig=AOvVaw0HFng41IOb_2MrOBasOj5j&ust=1531450449762596)
ADULT MALE ADULT FEMALE
1. _________________________ 1. _________________________
2. _________________________ 2. _________________________
3. _________________________ 3. _________________________
4. _________________________ 4. _________________________
5. _________________________ 5. _________________________
6. _________________________ 6. _________________________
7. _________________________ 7. _________________________
8. _________________________ 8. _________________________
9. _________________________ 9. _________________________
10. _________________________ 10. _________________________
11. _________________________ 11. _________________________
12. _________________________ 12. _________________________
13. _________________________ 13. _________________________
14. _________________________ 14. _________________________
15. _________________________
16. _________________________
ADULT MALE and FEMALE Strongyloides stercoralis

(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwj1hfDA55jcAhWFqJQKHfQGCTIQjxx6BAgBEAI&url=https%3A
%2F%2Fwww.slideshare.net%2FMizanRahman9%2Fstrongyloides-stercoralis-
63214733&psig=AOvVaw3CPcudMJa34md9a5dzZgJG&ust=1531458943943067)
ADULT PARASITIC FEMALE
1. ________________________________
2. ________________________________
3. ________________________________
4. ________________________________
5. ________________________________
6. ________________________________
7. ________________________________
ADULT FREE LIVING MALE
1. ________________________________
2. ________________________________
3. ________________________________
4. ________________________________
ADULT FREE LIVING FEMALE
1. ________________________________
2. ________________________________
3. ________________________________
4. ________________________________

Filariform and Rhabditiform larvae of Hookworms versus Filariform and Rhabditiform larvae of Strongyloides
stercoralis
(https://www.slideshare.net/PaemikaPidchayathana/nemathelminthes-review)
1. __________________________________________
2. __________________________________________
3. __________________________________________
4. __________________________________________

1. Describe the morphology of the ova and adult of class rhabditea
2. Describe the life cycle of: hookworms, S. stercoralis
3. Differentiate hookworm and S. stercoralis based on: mode of transmission, diagnostic and infective stage,
morphology
4. In a table form, differentiate filariform from rhabditiform larvae
5. List and describe the different laboratory methods for the identification of class rhabditea

NAME: RATING:
Experiment No. 8
PHYLUM NEMATODA - ORDER SPIRURIDA (BLOOD AND TISSUE NEMATODES)
I. Background
In Order Spirurida, six lips, lips absent, or lateral pseudolabia present surround the mouth. The
buccal capsule is usually well developed. The esophagus is divided into anterior muscular and posterior
glandular portions, never with bulb. The development from first to third-stage juvenile is in arthropod
intermediate hosts.
They are parasites in the intestine and deeper tissues of all vertebrate classes. The microfilaria
may be classified by the presence (sheathed) and absence (unsheathed) of a sheath. A sheath is a delicate,
close-fitting membrane that is derived from the original eggshell and is only detectable as its projects
beyond the head or tail.
The sheathed microfilarias are the following: Wuchereria bancrofti, Brugia malayi and Loa loa,
while the unsheathed microfilariae are Onchocerca volvulus, Dipetalonema perstans and Mansonella
ozzardi.

1. To be able to know the morphology of the ova and adult members Order Spirurida
2. To describe the life cycle of Blood and tissue nematodes
3. To know the mode of transmission for both Blood and Tissue nematodes
4. To be familiar with the diagnostic and infective stages of these Blood and Tissue nematodes.
5. Identify the nematodes using the distinguishing marks of their diagnostic stages including the
laboratory methods for identification

Microscope 2% formalin solution
Centrifuge tube Pipette
Prepared slides Colored plates
Reference Book Centrifuge

a. Procedure:
A. Knots Concentration Technique
1. Mix 2 ml blood with 10 ml of 2% formalin in the centrifuge tube
2. Centrifuge at moderate speed for 2 minutes
3. Decant the supernatant liquid
4. Pipette the sediment on the slide and examine under the microscope.
NOTE:
The sediment may be stained with a 1:10,000 aqueous solution of either methylene
blue or azure 11, and the sediment is examined under the microscope.
b. Draw and label:

a. Print and Label : Microscopic (HPO)
1. Group results
b. Print and Label: Diagrammatic
1. Posterior ends of sheathed and unsheathed microfilariae
2. Brugia malayi
PRINT and LABEL (diagrammatic)
MICROFILARIAE POSTERIOR END
Wuchereria bancrofti
Loa loa
Brugia malayi

Mansonella perstans
Mansonella ozzardi
Ochocerca volvulus
Mansonella streptocerca
ADULT Brugia malayi

ADULT Dracunculus medinensis
ADULT MALE and FEMALE Wuchereria bancrofti

(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwjz8sHmtZbcAhXGkZQKHWglAb
cQjxx6BAgBEAI&url=http%3A%2F%2Fwww.studyandscore.com%2Fstudymaterial-detail%2Fwuchereria-general-
characters-life-cycle-and-filariasis&psig=AOvVaw0fJE-gFf7d0yLP8M0dY0ea&ust=1531376764220625)
A. ADULT FEMALE B. ADULT MALE
1. _____________________ 1. ______________________
2. _____________________ 2. ______________________
3. _____________________ 3. ______________________
4. _____________________
5. _____________________

6. _____________________
7. _____________________
8. _____________________
9. _____________________
10. _____________________
11. _____________________
1. Describe the morphology of the ova and adult members of order Spirurida
2. Describe the life cycle and mode of transmission of blood and tissue nematodes
3. In a table form, differentiate the different members of order Spirurida based on diagnostic stage, infective
stage and distinguishing mark.
4. In a table form differentiate sheathed from unsheathed microfilaria
5. List and describe the different laboratory methods for identification of blood and tissue nematodes

PHYLUM NEMATODA – OTHER MEMBERS
NAME: RATING:
Experiment 9
I. Background
There are other members of Phylum Nematoda that are of equal importance with other
species in the Phylum since they can infect humans and may cause damage inside man’s body.
1. To be able to know the morphology of the ova and adult parasites
2. To describe the life cycle
3. To know the mode of transmission
4. To be familiar with the diagnostic and infective stages
5. To know the different laboratory methods applicable in their determination

Microscope
Prepared slides
Colored plates
Reference book
a. Print and Label: Diagrammatic
1. Toxocara canis ova and adult worm
2. Toxocara cati ova and adult worm
3. Gnathostoma pva and adult worm
4. Trichostrongylus ova and adult worm
5. Angiostrongylus cantonensis ova and adult worm
6. Anisakis ova and adult worm

NEMATODE OVA ADULT

Toxocara canis
Toxocara cati
Gnathostoma
Trichostrongylus
Angiostrongylus cantonensis
Anisakis
1. Complete the table below

NEMATODE Synonym/common Disease habitat Definitive host Infective
name stage
2. Illustrate through a flow chart the life cycle of the following nematodes
a. Toxocara canis
b. Toxocara cati
c. Gnathostoma
d. Trichostrongylus
e. Angiostrongylus cantonensis
f. Anisakis
3. List and briefly describe the different methods for the diagnosis of the following nematodes
a. Toxocara canis
b. Toxocara cati
c. Gnathostoma
d. Trichostrongylus
e. Angiostrongylus cantonensis
f. Anisakis

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF CESTODES
(TAPEWORMS)
1. identified each member of Phylum Cestoda according to morphology, and recognize

stage of the parasite being studied
LESSON PROPER
This part of the module consists of a discussion of the class of multicellular worms
noted for their flat or ribbon-like appearance known as Cestoda (the cestodes). The
characteristic appearance of the cestodes forms the basis for the common names
associated with this group, flatworms or tapeworms.
GENERAL CHARACTERISTICS
1. Elongated, ribbon-like, segmented, generally flattened dorsoventrically
2. No alimentary tract and vascular system
3. Food is absorbed by the cuticle
4. Parts
a) scolex: organ of attachment
b) neck: region of growth
c) strobila: chain of progressively developing proglottids
➢ Segments are either:

o Craspedote: proglottids overlap
o Acraspedote: proglottids don’t overlap
o Apolytic: segments are detached with mature eggs
o Anapolytic: proglottids are shed when they are exhausted of eggs
5. Excretory System has FLAME CELL/PROTONEPHRIDIUM

6. Reproductive System (most cestodes are MONOECIOUS and exhibit protandry)
• Vas deferens and vagina have common genital pore that may open on the following:
a) Ventral surface with uterine pore (D. latum)
b) Lateral: same side of proglottide (Hymenolepis species)
c) Lateral: irregularly alternate (Taenia spp)
d) Bilateral: 2 sets of reproductive organs are present (D. caninum)
• CIRRUS – male copulatory organ
• Male uterus
a) Coiled
b) Secular
c) Tube or straight
• Gravid uterus
a) Reticular with ova in capsules (D. caninum)
b) With lateral branches (Taenia spp)
TWO MAIN CLASSES OF LARVA
1. Solid
a) Procercoid- 2nd larval stage; bears 6 hooks near the posterior end
b) Plerocercoid- 3rd larval stage; has a solid body with a developing scolex & strobila
2. Vesicular or bladder or cystic
a) Cysticercoid – has a solid body with an invaginated scolex with poorly developed
bladder or no bladder
b) Cysticercus of true bladder- has a fluid filled and fully developed bladder
Variations:
a) Coenerus – has a well developed cyst with multiple invaginated scolices from
the germinal layer
b) Echinococcus/ hydatid cyst – has a well-developed cyst with brood capsules
and daughter cells with multiple scolices
COMPARISON OF THE TWO ORDERS
Points of Differentiation Pseudophyllidean Cyclophyllidean

1. Scolex Almond shape, with 2 with 4 muscular suckers
sucking grooves or
bothriaw
2. genital pore Center of the segment Margins of segment
3. Uterine pore Center of the segment none
4. Uterus coiled Sac-like; branched, eggs in
capsule
5. Ova Operculated, immature Non-operculated, mature
when laid when laid
6. Oncosphere Ciliated and called Non-ciliated but with 3
coracidium pairs of hook and called
hexacanth embryo
7. larval forms Solid Cystic
a) Procercoid a) cysticercoids
b) plerocercoid b) cysticercus
c) coenurus
d) hydatid
Order: PSEUDOPHYLLIDEAN
1. Diphyllobothrium latum
• Common name: Fish tapeworm/Broad tapeworm

• Diseases: Diphyllobothriasis, Fish or Broad tapeworm infection, Bothriocephaliasis
• 1st IH: copepods of Genus Cyclops and Diaptomus
• 2nd IH: Fresh water fishes like pike, salmon, trout, white fish
• Reservoir hosts: dogs and cats
• Final Host: man
• Infective stage to man: plerocercoid larva
• Distribution: cosmopolitan but prevalent in temperate zone where fresh water fish
forms integral part of the diet (Europe, lake region of Switzerland, North America, Asia)
• MORPHOLOGY:
1. Egg/ova

➢ 30-50 μm
➢ Single shelled, broadly ovoidal with the Posterior part thickened and with
operculum at one end.
➢ Yellowish to yellow-brown in color
➢ Immature if found in feces
➢ 100,000 eggs/day
2. Coracidium
➢ liberated from eggshell

➢ ciliated
➢ swims freely in the water
3. Procercoid larvae
➢ measures 550 in length

➢ with 3 pairs of hooklets
4. Plerocercoid larvae/sparganum
➢ Measures 6 mm in length
➢ Glistening, opaque white
➢ Unsegmented and has an invaginated anterior end
➢ No scolex
5. Adult
➢ Largest tapeworm of man
➢ Measures 3-10 mm in length with as many as 3, 000 proglottids
➢ Ivory white in color
6. Scolex
➢ Almond shaped, unarmed and has 2 deep dorsoventral suctorial grooves
7. Proglottids
➢ Mature segment is broader than long
• Pathology
➢ Systemic toxemia
➢ Mechanical obstruction of the intestines
➢ Bothriocephalus anemia of Tapeworm pernicious anemia
o Macrocytic normochromic anemia

o with thrombocytopenia
o mild leukopenia
➢ Intense local inflammation and eosinophilia
➢ Human sparganosis
➢ Ocular sparganosis – intense pain,irritation and edematous swelling of the eye
lids
• Laboratory Diagnosis
1. Examination of eggs and proglottids in feces
2. Kato thick smear preparation
3. Concentration technique
4. Surgical removal of the worms and drainage of the involved sites ( Human
sparganosis)
• Treatment
1. Praziquantel (5-10 mg/kg single dose)- Drug of choice
2. Niclosamide or Quinacrine hydrochloride- used to kill adult D. latum
3. Paramomycine
4. 40% ethyl alcohol with Procaine injected into the lesion to kill plerocercoid
• Prevention
1. Freezing (-18oC) for 24 hours or (-10oC) for 48 hours
2. Fish reservoirs should be kept free of raw sewage
3. Boiling and filtering of drinking water
4. De-worming of reservoir hosts
5. Thorough cooking of freshwater fishes
6. In endemic areas, feces should not be discarded into rivers and lakes

Order: CYCLOPHYLLIDEAN
1. Taenia solium
• Common Name: pork tapeworm
• Diseases: Taeniasis solium, Pork tapeworm infection, Cysticercosis
• Distribution: Cosmopolitan
• Morphology:
a) Ova- brown, spherical, 31-35 microns in diameter, with 2 radially striated shells
b) Adult- 2 to 7 meters in length
c) Scolex- globular with 4 suckers and armed with rostellum (circular tow of hooks)
d) Proglottid- 5 x 10 mm in size with 800-1000 proglottids
e) Gravid proglottid- contains 30,000-50,000 eggs; uterus exhibit 7-12 lateral branching

• Intermediate host: pig, man
• Pathology:
1. Taeniasis solium – caused by adult tapeworm
➢ Characterized by mild transitory intestinal obstruction and may give rise to
vague abdominal pain similar to hunger pains
➢ Caused by the ingestion of raw of insufficiently cooked pork containing
cysticercus cellulosae
2. Cysticercosis cellulosae – caused by the cysticercus cellulosae
➢ Caused by the accidental ingestion of T. solium, or thru regurgitation of eggs
and gravid segments of the parasite into the stomach
➢ Cysticercus cellulosae may lodge in vital organs like the brain, spinal cord,
heart, liver, eyes etc. giving rise to pressure symptoms
➢ The presence of growing larva provokes cellular reactions with blood cell
infiltration, fibrosis, necrosis of the capsule
➢ (60-90%) Neurocysticercosis – may cause epilepsy or disordered behavior,
intermittent obstructive hydrocephaly, disequilibrium, meningoencephalitis
and falling vision.
• Diagnosis:
a) Taeniasis: presence of ova or gravid proglottids can be observed through stool
examination; scotch tape swab
b) Cysticercosis: development of sub-cuticular nodules which provide material for
biopsy
• Treatment: Praziquantel, Bithionol, Panomomycin
• Prevention and Control
1. A thorough inspection of beef and pork
➢ adequate cooking or freezing
➢ Freezing meat at -20oC for 10 days
2. Avoid eating raw pork
3. Proper disposal of human feces
4. Good personal hygiene

2. Taenia saginata
• Common name: beef tapeworm
• Disease: Taeniasis saginata
➢ Disturbance in the normal functions of the digestive tract
➢ Systemic intoxication
• Intermediate host: cow
• Morphology
a) Ova- same with T, solium
b) Adult – whitish, opaque in color
- 5 to 10 meters in length
- each gravid proglottid contains 80,000 eggs
• Pathology
➢ Disturbance in the normal functions of the digestive tract
➢ Systemic intoxication- due to absorbed by-products of the worms
• Diagnosis
➢ Stool examination- DFS, KTS, Concentration Techniques
➢ Perianal Scotch Tape method
• Life cycle of T. saginata: same as T. solium except for the intermediate host
• METHODS OF DIFFERENTIATING TAENIA SPECIES
1. Examine the proglottids in the feces
➢ inject dye (India ink or safranin) using gauge 26 needle
2. Examine the scolex
➢ T. solium: armed with rostellum with 4 muscular suckers
➢ T. saginata: unarmed with 4 muscular suckers
COMPARISON OF TAENIA SPECIES
DIFFERENCES Taenia solium Taenia saginata

Common name Pork tapeworm Beef tapeworm
Disease/s Taeniasis solium Taeniasis saginata
CYSTICERCOSIS
Scolex Globular in shape with 4 Pyriform in shape with 4
cup-shaped suckers; muscular suckers; NO
rostellum and double rows hooklets
of hooklets
Mature proglottids Roughly square with Irregularly alternate lateral
unilateral or irregularly genital pore
alternate genital pore
Segments 800-1000 1000-2000
Testes 150-200 300-400
Uterine branches 7-12 15-30
Infective stage Cysticercus cellulosae Cysticercus bovis
- ellipsoidal, translucent, thin - pinkish cyst, opaque, with
walled bladder with an invaginated neck and scolex
opaque invaginated neck
and scolex equipped with
suckers and hooks
OTHER SPECIES
❖ Taenia saginata asiatica (Asian Taenia)

✓ MOT: eating raw pig liver; cattle and goat
✓ Morphology: scolex- with sunken rostellum, 2 rows of hooklets
✓ Asymptomatic, though multiple adult infection is possible- abdominal pain,
nausea, weakness, weight loss, headaches
❖ Taiwan taenia
✓ Adult scolex: unarmed
✓ IH: pig, cattle, goats, wild boars, monkeys
3. Hymenolepis nana
• CN: Dwarf Tapeworm

✓ Rats: H. nana var. fraternal
• IH: fleas, rice and flour beetles (Tenebrio spp)
• DH: man, rats, mice
• Infective stage:
✓ embryonated ova (Direct Life Cycle)
✓ cysticercoid larva (Indirect Life Cycle)
• Morphology
a) Ovum
✓ 45 µm in diameter; 4-8 “polar filaments”
✓ Size: 30-55 µm
✓ Oncosphere: six hooks (seen as dark lines at 8 o’clock)
b) Larva (Cysticercoid)
✓ Tailed structure w/invaginated scolex
✓ Lacks a fluid filled bladder
✓ Develops in insect or human villi (SI)
c) Adults
✓ Small, L: 15-50 mm
✓ Scolex: retractable rostellum w/20-30 Y-shaped hooklets
✓ Segments: wider than long, terminal gravid segments break up and release
eggs
✓ Genital pore: located on the same side
• Disease: Dwarf tapeworm infection
• Pathology
a) Generalized toxemia- due to the absorption of metabolic wastes of the parasites
b) Pruritis ani; intestinal obstruction; periodic diarrhea
c) Headache, dizziness, anorexia
• Diagnosis: Fecalysis
• Treatment:
a) Praziquantel (25mg/kg single dose): causes vacuolization and disruption of
tegument at the neck region
b) Niclosamide

• Control &
Prevention
➢ Preventing fecal
contamination of food
and water in institutions
and crowded areas
➢ General
sanitation: rodent and
insect control
(especially control of
fleas and grain insects)
➢ Treatment of
infected patients
➢ Personal hygiene

4. Hymenolepis diminuta
• Common Name: Rat Tapeworm

• Human: accidental infections
• Requires intermediate host
• Morphology
a) Ovum
➢ hexacanth larva (oncosphere)
- Enclosed in an inner membrane
- Lacks bipolar filaments
- Hooklets: fan-like arrangement
➢ D: 60-80 µm, bile stained
b) Adult
➢ L: 40-60 cm
➢ scolex is club-shaped: unarmed rostellum with four suckers
➢ Proglottids:
- L: 0.75 mm; W: 3.5 mm
- 3 ovoid testes and 1 ovary
- Genital pores: unilateral
- Uterus: sac-like with eggs
• Laboratory Diagnosis
➢ Fecalysis: eggs and whole worm
➢ Light infections: concentration techniques
• Treatment:
➢ Praziquantel : 25 mg/kg single dose
➢ Quinacrine
➢ Niclosamide
• Prevention and Control:
a) Preventing fecal contamination of food and water in institutions and crowded
areas

b) Protection of food especially the precooked cereals from insects
c) General sanitation; rodent & insect control
d) Treatment of infected patients
e) Personal hygiene
f) Sanitary disposal of human waste

Differentiation H. nana I. diminuta
Common name Dwarf tapeworm Rat tapeworm
Infection Dwarf tapeworm infection Hymenolepiasis diminuta
scolex Small, globular and bears Club-shaped, has
a short refractile rostellum rudimentary unarmed
with a single row of 20-30 rsotellum
small hooks
No of segments 200 800-1000
Mature proglottid Trapezoidal, 4x as broad as Same with H. nana
long with single genital

pore on its left side and 3
round testes and bilobed
ovary
Uterus Sacculate Sacculate, filled with egg
masses
Ova (has 6 hooklets) Has 4-8 polar filaments No polar filament
habitat Upper 2/3 of ileum S.I of rats and mice
Infective stage Embryonated ova Cysticercoid larvae
Intermediate hosts Flour fleas, flor beetles Insects, such as beetles,
cockroaches, rat fleas
Definitive host Man, rats, mice Man, rat
5. Dipylidium caninum
• Common name: Double-Pored Tapeworm/ Dog TW

✓ Very common parasite of dogs and cats worldwide
✓ Humans: accidental host
• Disease: Dipylidiasis or Dog tapeworm infection
✓ Intestinal disturbances including:
- indigestion
- loss of weight
- Epigastric pain
- Diarrhea
- Anal pruritus
- Allergic reactions
• Distribution: cosmopolitan in dogs and cats; human infection is raMorphologyre
• Morphology
a) Scolex- rhomboidal-shaped, with a club-shaped rostellum and thorn shaped hooks
b) Proglottids: (60-175 proglottids)
1. mature- pumpkin seed shaped with double set of reproductive organs
2. gravid- filled with polygonal-shape uterine egg pocket (8-15 effs in each)

3. ova – sub-
spherical to
oval, hyaline
with thick
transparent
albuminous
covering with
membranes;
each egg
contains an
oncosphere-
with 3 pairs of
delicate
hooklet
• Habitat:
Lumen of
small intestine
of dogs
• Infective stage: cysticercoid larvae
• Intermediate hosts: cat fleas (Ctenocephalides felis)
Dog fleas (Ctenocephalides canis)
Human fleas (Pulex irritans)
• Definitive host: dogs/canines
• Pathology: intestinal disturbances including indigestion and loss of weight
• Diagnosis: Stool exam (DFS, KTS, Concentration tech) observe for the eggs and gravid
segments
• Treatment: Quinacrine, Niclosamide, Bithionol
• Prevention: Children should be informed with the danger of playing with dogs
RAILLIETINA
• genus of parasitic tapeworms that has chicken, turkey, geese and numerous
other domestic and wild birds as final hosts.

• The most relevant ones are:
a) Raillietina cesticillus: the broad-headed tapeworm, up to 13 cm long
and 1-3 mm wide, in chicken, turkey and many wild birds; found
worldwide
b) Raillietina echinobothrida: the nodular tapeworm, up to 25 cm long and
1 -4 mm wide, in chicken, pigeon, pheasants; found worldwide
c) Raillietina tetragona: up to 25 cm long and 1-4 mm wide, in numerous
domestic and wild birds; found worldwide
d) Raillietina bonini: up to 7 cm long and 1.5 mm wide, mainly in pigeons;
found in Europe, Near and Middle East
Raillietina garrisoni
• Common intestinal cestode of rodents in the Philippines

• Morphology:
➢ L: 60 cm
➢ Scolex: minute; subglobular, has rostellum with 90-140 hammer shaped hooks
➢ Mature proglottid: bilobed, surrounded by 36-50 ovoid testes
➢ Genital pore: opens at the side
• IH: flour beetle (Tribolium confusum)
• Pathogenesis:
➢ Asymptomatic
➢ Medical consultation: proglottids in feces
• Diagnosis: Fecalysis – Proglottids and ova
• Treatment: Praziquantel: to expel the worm
• Prevention and Control:
➢ Elimination of rodents
➢ Proper storage of grain products
➢ Sanitary waste disposal
6. Multiceps multiceps/ Taenia multiceps
• Common name: “Gid worm”

• Intermediate host: herbivores

• Definitive host: dogs, wolves and fox
• Infective stage: embryonated eggs
• Disease: cerebral & ocular coenurosis (coenurosis- BLADDER WORM)
• Adult: 40-60 cm
➢ Scolex: pyriform with double circles of rostellar hook
➢ Uterus: 18-20 branches
• Treatment: Praziquantel- to expel the worm
• Prevention and control

➢ Elimination of rodents
➢ Proper storage of grain products
➢ Sanitary waste disposal
7. Echinococcus granulosus
• CN: Hydatid Worm, Hydatid cyst

• Intermediate host: sheep, cattles, horses
• Definitive host: dogs and other canines
• Disease: unilocular echinococcosis, hydatid disease, echinococciasis
❖ Man is parasitized only by the LARVA (hydatid cyst) of the tapeworm. The dog is
the optimum definitive host which consumes the viscera of the IH containing
the infective larval stage and thereby becomes infected.
• Geographical distribution: Australia, New Zealand, South Arica and Africa
• Morphology:
a) Eggs: cannot be differentiated from the eggs of Taenia
b) Adult: shortest tapeworm with only 3 segments; 3-9 mm in length
SCOLEX: Pyriform with 4 suckers and armed rostellum (double crown of 20-36
hooklets)
PROGLOTTIDS: Immature, mature and gravid proglottid
✓ Gravid segment- uterus with its lateral invagination resemble a loosely
twisted coil. Uterus has 12-15 branches distended with around 500 eggs.
c) Larva: hydatid cyst
TYPES OF HYDATID CYST
1. Unilocular cyst- consists of laminar and germinal layers, most prevalent human
type of hydatid cyst; may grow for 5-20 years before it is diagnosed
2. Alveolar cyst- lacks the laminar layer which is the mother cyst
3. Osseous- lacks the laminar layer; found in the bones
• Pathogenesis
Damage produced by the larval forms are mechanical and toxic. Young cyst may
lodge in vital organs like the capillary beds of brain or heart valves producing
dangerous obstruction. In the bone, they cause erosion leading to fracture. In the
abdomen, they grow into tremendous size and burst, followed by anaphylactic
reactions caused by the toxic hydatid fluid
Liver (66%)
- Obstructive jaundice
- Fever
- eosinophilia
Lungs (22%)
- Coughing with allergic symptoms
- Sputum: frothy blood, mucus, hydatid fluid
Kidneys (3%)
- Intermittent pain
- Hematuria
- Kidney dysfunction
Brain
- Increased intracranial pressure

- Jacksonian epilepsy
• Diagnostic stage: Larval stage
a) X-ray (Roentgenography): for hydatid cyst in the lungs, thoracic involvement,
extrahepatic abdominal cyst and in the long bones femur)
b) Hydatid thrill: vibrations felt which is a special diagnostic sign in unilocular hydatid
cyst of the abdominal viscera
c) Exploratory cysts puncture
d) CBC-eosinophilia
e) Immunologic tests
➢ Bentonite flocculation test
➢ Casoni’s test intradermal test
➢ FAT, IHA, ELISA, Immunoelectrophoresis
• Treatment
a) Surgical removal of the cyst; Replacement of the cyst fluid with 10% formalin or 2%
AgNO3
b) Albendazole: 400 mg 2x a day for 4 weeks
c) High dose of Mebendazole
• Prevention
a) Personal hygiene
b) Deworming of dogs
c) Safeguarding all discarded viscera in slaughter houses by boiling or dumping into
dog proof pits
8. Echinococcus multilocularis
• Synonym: Echinococcus alveolaris

• DH: fox, canines, coyotes, cats
• IH: rodents and sometimes humans
• MOT: Fecal-oral
➢ eating raw plants contaminated w/ DH’s feces
• Disease: Alveolar Hydatid disease
• Distribution: known in Europe, Asia, New Zealand, South and North America
• Morphology & Life cycle: similar to E. granulosus
➢ Adult: 3-5 segments (1.2-3.7mm)
➢ Cyst: multilocular
• Pathogenesis: varies, some infections may spontaneously disappear, others go till
death
• Treatment
➢ Surgery
➢ High doses of Albendazole- resistant to praziquantel

SYNTHESIS
Cestodes, or tapeworms, include multiple species of flat worms that can reside in the
human gastrointestinal tract. The species that most commonly cause human disease include
Taenia saginatum, Taenia solium, Diphyllobothrium latum and Hymenolepis nana among
others.
ASSESSMENT
SEATWORK: Complete the table below
Infective stage Disease caused

Fish tapeworm
Pork tapeworm
Beef tapeworm
Dwarf tapeworm
Rat tapeworm
Dog tapeworm
Gid worm
Hydatid worm
Alveolar cyst
QUIZ
or google forms.
REFERENCES
Encyclopedia.com. (2020) Secernentea (Secernenteans) Environment Encyclopedias

almanacs transcripts and maps Secernentea (Secernenteans) Retrieved from

https://www.encyclopedia.com/environment/encyclopedias-almanacs-transcripts-
and-maps/secernentea-secernenteans on July 12, 2020


LABORATORY
NAME: RATING:
Experiment No. 10
PHYLUM PLATYHELMINTHES - CLASS CESTODA
I. Background
The cestodes are flatworms, ribbon-like in shape and are markedly segmented. Adults live
attached to the intestinal wall through an organelle of attachment called scolex. They are not
provided with a digestive system and food is absorbed form the host's intestines. They are
hermaphroditic, so each segment has the male and female reproductive system. The segments
are also called proglottids and the chain of segments is called strobila. Segments that are near the
neck are immature followed by mature segment and the most distal are the gravid segments,
which have fully developed uterus, filled with eggs.
For the identification of cestodes, eggs, proglottids and scolex are important criteria used
in the laboratory.

1. To be able to know the morphology of the ova and adult members class Cestoda
2. To describe the life cycle of Human Tapeworms
3. To know the mode of transmission of tapeworms
4. To be familiar with the diagnostic and infective stages of these tapeworms.
5. Identify the tapeworms using the distinguishing marks of their diagnostic stages
6. To know the morphology and laboratory methods applicable in the determination of
Cyclophyllidean and Pseudophyllidean
Microscope
Prepared slides
Colored plates
Reference Book

a. Print and Label: Microscopic
1. Pseudophyllidean
a. ova of Diphyllobothrium latum (HPO)
b. segment (Scanning objective)
2. Cyclophyllidean
a. Ovum of Taenia species (HPO)
b. Taenia solium scolex (Scanning objective)
c. Taenia solium cysticercus cellulosae (scanning objective)
d. Taenia solium proglottid (scanning objective)
e. Hymenolepis nana ova
f. Hymenolepis diminuta ova
g. Dipylidium caninum ova
h. Echinococcus granulosus (whole mount)
1. Taenia solium scolex
c. Print and Label: Diagrammatic
1. Diphyllobothrium latum scolex
2. Raillietina garrrisoni ova and adult worm

ova of Diphyllobothrium latum (HPO) segment (Scanning objective)
Ovum of Taenia species (HPO) Taenia solium scolex (Scanning objective)
Taenia solium cysticercus cellulosae Taenia solium proglottid (scanning objective)
(scanning objective)

Hymenolepis nana ova Hymenolepis diminuta ova
Dipylidium caninum
ova Echinococcus granulosus (whole
mount)
Taenia solium scolex
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwikq86Gv5bcAhUFnJQK
HXXWAgYQjxx6BAgBEAI&url=http%3A%2F%2Fwww.biologydiscussion.com%2Fparasitology%2Fparasitic-
worm%2Ftaenia-solium-linnaeus-history-and-functions%2F62216&psig=AOvVaw0rKUyJTtzd-
zjr6uFLpYIU&ust=1531379404017475)
1. _____________________________
2. _____________________________
3. _____________________________

4. _____________________________
5. _____________________________
6. _____________________________
7. _____________________________
8. _____________________________
9. _____________________________
Diphyllobothrium latum scolex
Raillietina garrrisoni ova Raillietina garrrisoni adult worm

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF TREMATODES
(FLUKES)
1. In a table form, describe the morphology of the ova and adult members of class Cestoda
CLASS CESTODA OVA ADULT
2. Describe the following:

a) Life cycle of cestodes
b) Mode of transmission of cestodes
3. In a table form, differentiate Cyclophyllidea and Pseudophyllidea
4. Differentiate the different members of class Cestoda based on common name, disease caused,
mode of transmission, diagnostic and infective stage, distinguishing mark/s
1. identified each member of Phylum Trematoda according to morphology, and

2. Valued the importance of ethics of observing patients’ and results’
confidentiality
LESSON PROPER
This part of the module consists of a discussion of the class of helminth parasites
belonging to the class known as Trematoda or Digenea. Like the cestodes, the
trematodes belong to the phylum Platyhelminthes. For classification purposes, the flukes
may be divided into four categories based on the areas of the body that primarily
harbor the parasites: intestinal, liver, lung, and blood. Commonly known as the flukes,
these parasites vary in egg, larva, and adult morphology and production processes.

The trematodes can be divided into two groups, the hermaphroditic (self-fertilizing)
flukes that infect organs and are foodborne, and the blood flukes or schistosomes that
are dioecious (parasites that reproduce via separate sexes) and infect by direct
penetration. Common to all trematodes is their complex life cycle, which almost always
include mollusks (snails) as an intermediate host.
1. Morphology: Broadly oval/leaf-like, bilaterally symmetrical along a long axis and

are usually flattened dorsoventrally, EXCEPT for the blood flukes which are
cylindrical; shape and size varies.
2. Excretory system: bilaterally symmetrical and open at posterior end with FLAME
CELLS or SOLENOCYTES
3. Digestive system:
• With an oral sucker and a globular pharynx extending from the mouth to
a short narrow esophagus
• Oral cavity is situated at the anterior end of the worm and is equipped
with muscular suckers with spines or hooklets. Some species possess a
large ventral sucker (ACETABULUM) posterior to the oral sucker
• Esophagus bifurcates in front of the ventral sucker into a pair of blind
intestinal caeca which may be simple or branched or may reunite to form
a single caecum
• Digestion is predominantly an extra-cellular process
4. Nervous system: composed of a group of paired ganglion cells disposed like
saddle on the dorsum of pharynx/esophagus
5. Reproductive system:
• All are hermaphrodite/monoecious EXCEPT the blood flukes
• All are Oviparous
• Eggs are provided with a lid called operculum, EXCEPT the blood flukes
• The male reproductive system is located at the posterior half of the body.
It consists of the prostate gland, seminal vesicle, cirrus, vas deferens, vas
efferens, and testes

• The female reproductive system is composed of uterus, mehli’s gland,
ootype, vitellaria, seminal receptacle, and ovary.
6. Movement is either through contraction, elongation or flexion
7. Life cycle notes:
• The trematodes need body of water to complete their life cycle
• All require 2 intermediate hosts EXCEPT for blood flukes
• All trematodes lay eggs which upon embryonation, give rise to a ciliated
embryo known as MIRACIDIUM →CERCARIA (Dioecious flukes) →
METACERCARIA (Monoecious flukes)
8. Stages of Trematodes:
a. Egg stage
b. Larval stages
▪ Miracidium – a ciliated embryo that hatches in water
▪ Mother sporocysts – sac like structure
▪ Rediae/daughter sporocysts
▪ Cercaria/daughter redia – tailed and free swimming
▪ Metacercaria – the encysted resting or maturing stage
c. Adult stage
Morphology:
▪ Testes types:
✓ Branched, arranged in tandem: F. hepatica, C. sinensis, F.
buski
✓ Lobulated, arranged side by side: P. westermani
✓ Deeply lobulated, arranged in tandem or dumbbell testes:
E. ilocanum
✓ Oval or round, arranged side by side or slightly oblique: H.
heterophyes, M. yokogawai
✓ Lobular, obliquely arranged: O. felineus
✓ Oval, round: M. yokogawai
▪ Ovary types:
✓ Branched: F. hepatica, F. buski
✓ Lobulated: C. sinensis, O. felineus, P. westermani

✓ Oval or rounded: E. ilocanum, H. heterophyes, M
yokogawai
▪ Vitellariae types:
✓ Highly branched: F. hepatica, C. sinensis
✓ Finely granular vitelline follicles: P. westermani, F. buski
✓ Medium-sized vitelline follicles: E. ilocanum
✓ Compressed vitelline follicles: O. felineus
✓ Polygonal: H. heterophyes, M. yokogawai
▪ Intestinal ceca types:
✓ Branched: F. hepatica
✓ Simple: F. buski, E. ilocanum, M. yokogawai, H. heterophyes,
C. sinensis, O. felineus
✓ Simple but arranged in zigzag fashion: P. westermani
▪ Cercaria types:
o Lophocercus:
❖ Simple tailed: F. hepatica, F. buski, E. ilocanum
❖ Keeled-tail: C. sinensis, O. felineus, H. heterophyes
o Microcercus: P. westermani
o Forked tail: Schistosoma
Differences Between the Monoecious & Dioecious Flukes

Monoecious Dioecious
1. Type and number of hosts 2 intermediate hosts: 1 intermediate host
▪ Snails ▪ Snails only
▪ Fresh water
vegetations
2. Infective stage Metacercariae Cercariae
3. Ova Operculated Non-operculated with spines
4. Cercaria Straight-tailed Forked or bifurcated
5. Adult Flat, leaf-like Cylindrical, separate sexes
(hermaphrodites)
6. Habitat Lungs, liver, and intestines Blood vessels
Classification of Trematodes According to Habitat:

(DIOECIOUS FLUKES)
1. Liver flukes: c. Heterophyes heterophyes

a. Fasciola hepatica d. Metagonimus yokogawai
b. Fasciola gigantica 3. Lung fluke:
c. Clonorchis sinensis a. Paragonimus westermani
d. Opistorchis felineus 4. Blood flukes:
e. Dicrocoelium dendriticum a. Schistosoma japonicum
2. Intestinal flukes: b. Schistosoma mansoni
a. Fasciolopsis buski c. Schistosoma
b. Echinostoma ilocanum haematobium

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LESSON PROPER
The dioecious flukes are considered as the most important digenetic parasites of
man inhabiting the veins of their vertebrate hosts. The usual portal of entry into the
definitive host is the skin. Members of this group parasitize fishes, turtles, birds, and
mammals. There are at least four species (S. haematobium, Schistosoma mansoni,

Schistosoma japonicum, and S. mekongi) that are important agents of disease in man,
whereas others are incidental or potential human parasites. Species parasitizing man
usually inhabits the mesenteric-portal and caval venous blood vessels.
1. Morphology: The adult male and female

typically live in PAIRS
▪ Males:
➢ Larger than the female adult
worm measuring about 12-20 mm
by 0.5 to 0.55 mm in diameter
➢ Grayish in color and is attached
by its suckers to the wall of the
vessel
Source:
➢ With a gynecophoral canal on https://www.cdc.gov/dpdx/schistosomiasis/imag
es/7/S_mansoni_adult_Lammie1.jpg
the ventral side which functions to
surround almost completely the female adult facilitating copulation
▪ Females:
➢ Average size is 26 mm in length by 0.3 mm in diameter
➢ Thread-like structure
2. Infective stage: CERCARIA
3. Distribution: China, Japan, Indonesia, Thailand, Cambodia, Philippines
LIFE CYCLE:

1. Once the eggs are passed out in fresh water, the miracidium is released from the
egg and must locate a snail (intermediate host), where it develops into
cercariae
2. The miracidia in snail develops into primary sporocyst, then to a secondary
sporocyst that develops into cercaria
3. The cercaria will then escape into the water in 1-3 days and is ready to infect a
mammalian host
4. Human infection (Schistosomiasis) occurs in fresh water following the penetration
of fork-tailed cercaria into the skin of a host

5. Once it enters the definitive host, the cercaria will develop into schistosomula
and migrates into the portal veins, where maturation into adulthood is
completed.
6. The location of the adult flukes varies by species: S. mansoni and S. japonicum
reside in the veins that surround the intestinal tract, as well as in the blood
passages of the liver; while S. haematobium resides in the veins surrounding the
urinary bladder.
MAJOR SPECIES OF SCHISTOSOMES:
A. Schistosoma japonicum
▪ Common name:
o Oriental blood fluke
o Visceral blood fluke
▪ Main habitat:
o Superior mesenteric veins
▪ Morphology:
o Ova:
➢ Measuring about 50-85 um
by 38-60 um;
➢ Oval to sub-spherical with
minute lateral spines/knob
o Adult: Source:
https://www.cdc.gov/dpdx/schistosomiasis/images
➢ Male: /3/S_japonicum_egg_2X2b.jpg
➢ shorter but sturdier; measuring about 12-20 mm by 0.4-0.5 mm

➢ with 7 ovoidal testes arranged in single row behind acetabulum
➢ union of ceca and testes is posterior to the middle half of the
body
➢ Female:
➢ More delicate than males
➢ Longer and slender
➢ With pyramid ovary located at the center of the body (containing
50-100 eggs)

▪ Intermediate hosts: (Snails)
o Oncomelania quadrasi (Philippines)
o Oncomelania nocophara (Japan)
o Oncomelania formosana (Taiwan)
▪ Disease:
o Katayama disease
o Oriental schistosomiasis
o Visceral schistosomiasis
▪ Distribution: China, Japan, Indonesia, Thailand, Cambodia, Philippines
B. Schistosoma haematobium
▪ Common name:
o Vesical blood fluke
o Bladder fluke
▪ Main habitat:
o Veins of the urinary bladder
▪ Morphology:
o Ova:
➢ Ellipsoidal with prominent terminal
spines measuring about 110-170 um
by 38-70 um Source:
https://mcdinternational.org/trainings/
malaria/english/dpdx5/IMAGES/Parasit
eImages/S-
Z/Schistosomiasis/S_haematobium_egg
o Adult: _2X2a.jpg
➢ The ovary is located at the posterior to middle of the body

containing 20-30 eggs
▪ Intermediate hosts: (Snails)
o Biomphalaria (Africa)
o Bulimus (Middle east)
▪ Disease:
o Vesical/Urinary schistosomiasis
o Schistosoma hematuria

▪ Distribution: Africa, Western Asia, Southern Europe
C. Schistosoma mansoni
▪ Common name:
o Manson’s blood fluke
▪ Main habitat:
o Inferior mesenteric veins
▪ Morphology:
o Ova:
➢ Ellipsoidal with prominent lateral
spines
➢ Measures 112-182 um by 40-75 um
➢ Appear in the feces 30-40 days
after infection
o Adult:
➢ Male:
➢ Union of ceca is at the middle half Source:
https://blogs.biomedcentral.com/bug
of the body containing 8-9 testes bitten/wp-
content/uploads/sites/11/2019/04/Sc
➢ Female: histosoma_mansoni_egg_4841_lores-
620x342.jpg
➢ Considered as the smallest
schistosome
➢ Ovary is at the anterior to the middle half of the body containing
1-4 eggs
▪ Intermediate hosts:
o Biomphalaria (Africa)
o Tropicorbis (South America)
o Austrolarbis
▪ Disease:
o Intestinal Bilharziasis
o Schistosomal dysentery
▪ Distribution: Africa, Arabian Peninsula, Brazil, Venezuela

PATHOPHYSIOLOGY:
There are three (3) progressive stages in the pathologic processes of Schistosomiasis:
1. Pre-patent/Developmental stage:
➢ Starts from the penetration of the skin to the arrival of the mature worm to
the venules of the intestines or urinary bladder where female adults are
ready to oviposit
➢ Pathologic changes that occur:
✓ Negligible to mild cutaneous lesions at the side of cervical entry
✓ Tissue reactions to the immature worms inside and outside the
blood vessels
✓ Some associated toxic and allergic reactions
2. Acute stage:
➢ There is active egg deposition and extrusion
➢ Symptoms include: Loss of weight and relapse of pain
3. Chronic stage:
➢ Stable egg output and tissue proliferation
➢ Invasion of the tissues by the migrating larva
➢ Symptoms include:
✓ Petechial hemorrhage
✓ Small foci of eosinophilic and neutrophilic infiltration in the lungs
with cough and hemoptysis
✓ Acute inflammation reactions in the liver with fever and urticaria
LABORATORY DIAGNOSIS:
1. Demonstration of eggs in stool sample is diagnostic of Schistosomiasis

2. Concentration techniques – useful in chronic and light infections
3. Liver biopsy/Proctoscopic aspiration
4. Diagnosis of S. japonicum and S. mansoni
➢ Demonstration of eggs in feces
➢ Rectal biopsy

5. Diagnosis of Urinary schistosomiasis:
➢ Demonstration of eggs in urine or recovery of eggs through bladder
biopsy
➢ Accompanied with dysuria and hematuria
6. Immunodiagnostic Tests:
o COPT (Circumoval Precipitin Test)
➢ Principle: it is based on patient serum precipitation with lyophilized
eggs identified under microscope
➢ Uses lyophilized eggs to detect antibodies in patient serum
➢ Gold standard test for diagnosis of Schistosomiasis
➢ Method of choice and is routinely used in the Philippines as
diagnostic test for schistosomiasis
➢ DISADVANTAGE: Cross-reactivity with other helminths (affects
specificity)
➢ Procedure:
a. One drop (0.025 mL) of
the suspension containing
larvae, eggs, or immature
adults are put into the
well of a slide and 3 drops
(0.075 mL) of serum is
Source:
added https://www.researchgate.net/profile/Ta
b. A cover slip is placed over mer_Elbaz/publication/228816902/figure
/fig13/AS:650767220342785@153216640
the well 4890/Schistosoma-Circumoval-precipitin-
test-CPT.png
c. Incubated at 34°C for 24
hours
d. After incubation, the slide is examined under the
microscope
e. Positive result: appearance of precipitates/bleb formation
attached to the worms or eggs
o ELISA – highly sensitive test

TREATMENT:
✓ Praziquantel
✓ Oxamniquine
✓ Metrifonate
✓ Niridazole
PREVENTION AND CONTROL:
✓ Snail control
✓ Sanitary disposal of human excreta
✓ Treatment of infected individual
OTHER SCHISTOSOMES:
S. mekongi (Mekong schistosome) – similar with S. japonicum

S. bovis – a common parasite of cattle, sheep, goats, and equines
S. mattheei
S. curassoni
S. intercalatum
S. incognitum
SYNTHESIS
The schistosomes of man are similar in their basic life cycles and in their
pathologic effects on mammalian host tissues, yet they differ in morphology of the
adults, the shape of their eggs and larvae, in the groups of snails that they utilize as an
intermediate host, and in host susceptibility. The most significant species of Schistosoma
that causes disease to humans are S. japonicum, S. mansoni, and S.
haematobium.
ASSESSMENT
SEATWORK: Complete the table below (15 POINTS).
S. japonicum S. haematobium S. mansoni
Common name

Habitat
Appearance of ova
Intermediate hosts
Disease
QUIZ
✓ For offline and weak connectivity, it will be provided by the instructor in essay
form
✓ For Online/Strong connectivity, it will be provided either through a worksheet, or
quizziz or google forms.
REFERENCES
Beaver, P. (1984). Clinical Parasitology (9th edition). JMC Press Inc.
Zeibig, A. (2013). Clinical Parasitology: A Practical Approach. (2nd ed.). Saunders Elsevier
Inc.
Medina (c.m). COPT Technique. Retrieved from

https://www.slideshare.net/anjeanlopez/copt-technique on July 27, 2020
Nsengiyumva (e.n). Diagnostic Procedures of Schistosomiasis. Retrieved from

https://www.slideshare.net/EmmanuelNSENGIYUMVA3/diagnostic-procedures-of-
schistosomiasis on July 27, 2020
LABORATORY
NAME: RATING:

Experiment No. 11
PHYLUM PLATYHELMINTHES - CLASS TREMATODA (DIOECIOUS FLUKES)
I. Background
Trematodes or flukes, like all members of phylum platyhelminthes, are dorsoventrally
flattened, bilaterally symmetrical helminthes without a body cavity (acoelomate). They differ from
the cestodes in their gross morphology and manner of development. Unlike cestodes, flukes have
a digestive tract. The predominantly leaf-shaped worms characteristically possess suckers
prompting early researchers to call them Trematoda meaning, "body with holes".
All trematodes are heteroxenous (have more than one host) except for Schistosomes
(homoxenous). Unlike cestodes, trematodes never used humans as intermediate host. As a rule,
the first intermediate host is almost always a snail. One or more sexually reproducing embryonic
stages collectively referred to as polyembryony characterizes the trematode life cycle.

1. To be able to know the morphology of the ova and adult members class Trematoda
2. To describe the life cycle of Dioecious flukes
3. To know the mode of transmission Dioecious flukes
4. To be familiar with the diagnostic and infective stages of Schistosomes
5. Identify the Schistosomes using the distinguishing marks of their diagnostic stages

Microscope
Prepared slides
Colored plates
Reference Book
A. Procedure:
a. CIRCUMOVAL PRECIPITIN TEST (COPT)
The COPT is routinely performed in parallel to the Kato-Katz stool examination technique.
It was first developed for the diagnosis of schistosomiasis mansoni. COPT has been reported
to be a very sensitive assay, detecting more than 95% of individuals harboring current S.
japonicum infection.
1. A small amount of freeze-dried egg is placed on a microscope slide
2. Place a drop of the patient's serum.
3. Place a coverslip on top of it and seal the four sides with paraffin.
4. Incubate the specimen at room temperature or in an incubator for 24 to 48 hours.
5. Examine under the microscope for the presence of COP reactions around the eggs.
b. Kato-Katz Method or the Cellophane Covered Thick Smear

This procedure uses a measured amount of stool, which has been sieved through a wire
mesh and pressed under a cellophane paper soaked in glycerin-malachite green solution. A

uniform amount of stool is examined through the use of a template with a uniform-sized hole
in the middle. All eggs are counted from the whole preparation. The total egg count is
multiplied by a factor depending on the amount of stool used.
The procedure is useful for assessing the intensity of infection in schistosomiasis and
common soil-transmitted helminthiasis like ascariasis. trichuriasis and hookworm infection.
Consistency of the stool is the main determinant for the sensitivity of this technique; since
drier stools yield higher egg counts than moist ones. The technique can only be done on fresh
stools and not on liquid preserved samples.
For the identification of Schistosoma ova, 1% eosin solution can be layered over the
cellophane paper. This method can help in the visualization of the miracidium.
B. Draw and label:

a. Print and Label: Microscopic (HPO)
1. ova of Schistosoma japonicum
2. ova of Schistosoma mansoni
3. ova of Schistosoma haematobium
4. cercaria of Schistosomes
1. adult male and female S. japonicum in copula
2. adult male and female S. mansoni in copula
3. adult male and female S. haematobium in copula
ova of Schistosoma japonicum ova of Schistosoma mansoni

ova of Schistosoma haematobium cercaria of Schistosomes
LABEL (Diagrammatic)
(A. http://www.sfu.ca/biology/courses/bisc318/2015%20pdfs/Lecture_18_Feb%2024_Shisto.pdf)
A. adult male and female S. haematobium in copula
B. adult male and female S. mansoni in copula
C. adult male and female S. japonicum in copula
1. ________________________________ 7. _______________________ 13. _______________

2. _________________________________ 8. _______________________
3. _________________________________ 9. _______________________
4. _________________________________ 10. ______________________

5. _________________________________ 11. ______________________
6. _________________________________ 12. ______________________
1. Describe the morphology of ova and adult members of class trematode

2. Describe the following:
a) Life cycle of dioecious flukes
b) Mode of transmission
c) Diagnostic & infective stage as well as distinguishing marks of Schistosomes
d) In a table form, differentiate the different species of Schistosomes
3. List and describe the laboratory methods to identify dioecious flukes

(MONOECIOUS FLUKES)

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LESSON PROPER
The trematodes pass through three morphologic forms during their life cycle –
eggs, multiple larval stages, and adult worms. The eggs, which are the primary
morphologic form recovered in human specimens, vary in appearance. Some contain
a lid-like structure called operculum that under appropriate conditions, flips open to
release its contents for further development, such as in Fasciolopsis and Fasciola.
Based on the organism’s life cycle, the trematodes may be placed into two
categories, those that reside in the intestine, bile duct, or lung (organ dwelling). Human
infection of such organ-dwelling flukes occurs following ingestion of water plants, fish,
crab, or crayfish contaminated with encysted form of the parasite known as
metacercaria.
I. Liver Flukes
A. Family Fasciolidae
These large digenetic trematode species belong to the family Fasciolidae.
They are parasites found in the liver and biliary passages of humans and
herbivorous mammals. Fasciola hepatica (temperate liver fluke) and Fasciola
gigantica (tropical liver fluke) are the causative agents of fascioliasis.
The mode of transmission of Fasciola hepatica and Fasciola gigantica is
through ingestion of metacercaria encysted on edible water plants or by drinking
water contaminated with metacercaria. Thus, the life cycle of Fasciola hepatica
and Fasciola gigantica is the same.
LIFE CYCLE:
1. Upon ingestion, the metacercaria excysts in the duodenum or jejunum, liberating
the juvenile fluke
2. This juvenile fluke will then penetrate the intestinal wall to reach the peritoneal
cavity where it wanders over the viscera until it reaches the liver
3. The parasite then burrows through the liver parenchyma, feeding and growing,
until it finally enters the bile ducts where it becomes sexually mature in 3-4
months (the life span of the adult worm is 9-13 years)
4. Immature eggs are carried by the bile into the intestine and subsequently passed
out in the feces
5. The eggs will undergo maturation in the water forming a viable miracidium

6. The miracidium escapes through the operculum of the egg to seek out and
infect the 1st intermediate host (snail).
7. Inside the snail, the miracidium develops into sporocyst, followed by one or two
generations of rediae which produce cercariae.
8. After escaping from the snail host, the cercaria swims freely in the water, to seek
out for its 2nd intermediate host (aquatic plants)
9. Once it reached its 2nd IH, it will detach its tail and encysts in surfaces of aquatic
plants forming a metacercaria (infective stage)
a. Fasciola hepatica
▪ Common name: Sheep liver fluke
▪ Distribution: found in areas where sheep and cattle are raised and where
humans consume raw watercress (Europe, Middle East and Asia)
▪ Morphology:
o Adult:
➢ Large broad and flat body
which measures about 20-50
mm in length and 6-12 mm in
width
➢ A distinguishing feature is the
cephalic cone which has a
marked widening at the base
of the cone (“shoulder”).
➢ Testes: tandem formation and
highly branched
➢ Vitellaria: branched, found in
Source:
lateral and posterior portion https://www.cdc.gov/dpdx/fascioliasis
/images/2/F_hepatica_adult_stained_
➢ Ovary: branched/dendritic;
BAM.jpg
located posterior to uterus and
anterior to testes

➢ Uterus: short and coiled
o Ova:
➢ Large and ellipsoidal
➢ Thin-shelled with distinct
operculum
➢ Yellowish to light brown
in color
Source:
➢ Immature when laid https://www.cdc.gov/dpdx/fascioliasis/images/1/F_
hepatica_egg_wtmt_HB1.jpg
▪ Infective stage: METACERCARIA

▪ 1st IH:
o Water snail (Lymnea)
▪ 2nd IH:
o Ipomea obscura (kangkong)
o Nasturtium officinale (water cress)
▪ Disease and Clinical Manifestations:
o Liver rot disease/Fascioliasis hepatica
o Acute phase: (migration of the immature fluke through the hepatic
parenchyma)
➢ Abdominal pain, diarrhea, vomiting
➢ Hepatomegaly, fever, urticaria
➢ Eosinophilia
o Chronic phase: (adult fluke within the bile ducts)
➢ Intermittent biliary obstruction and inflammation
➢ Ectopic locations of infection (intestinal wall, lungs,
subcutaneous tissue, and pharyngeal mucosa)
▪ Laboratory diagnosis:
1. Direct fecal smear examination
2. Concentration and sedimentation techniques
3. Antibody detection

▪ Treatment:
1. Bithionol (treatment of choice)
2. Emetine hydrochloride
▪ Prevention and Control:
1. Thorough washing or cooking of vegetable
2. Boiling of water in areas where the infection is endemic
3. Eradication/elimination of the snail intermediate hosts
b. Fasciola gigantica
▪ Common name: Giant Liver Fluke
▪ Distribution: Infections with F. gigantica have been reported, more rarely in
Asia, Africa, and Hawaii
▪ Morphology: Same with Fasciola hepatica EXCEPT for the following:
o Greater in length, shorter cephalic cone

o Larger ventral sucker
o Reproductive organs are located on the anterior portion of the worm
o The ovum is bigger
o Adult:
➢ Compared to F. hepatica, the F.
gigantica adult worm is longer
and its “shoulder” (cephalic cone)
is less developed
➢ Ceca are more branched
➢ Branches of ovary are longer and
more numerous
Source:
https://assets.fishersci.com/TFS
-Assets/CCG/product-
images/F73642~p.eps-650.jpg

o Ova:
➢ The ova of F. hepatica and F.
gigantica are similar although F.
gigantica egg is bigger
▪ 1st IH: Water snail (Lymnea)
▪ 2nd IH: Ipomea obscura (kangkong) and
Nasturtium officinale (water cress) Source:
https://lh3.googleusercontent.com/pro
▪ Disease and Clinical Manifestations: xy/AsMbyJyrcFYaDgd6zilWi_tSwVHJ5R
kO1T2s1lVzcj9XH4gZLxKzBnLZP_onc3u
o Fascioliasis nk8vLRjsp3lWzyKZ2Mha8go6YrYYY8l8x
ExuZX5MIocs8MK4
▪ Laboratory diagnosis: Same with F. hepatica
▪ Treatment: Same with F. hepatica
▪ Prevention and Control: Same with F. hepatica
B. Family Opisthorchiidae
These small digenetic trematodes (C. sinensis, O. felineus, O. viverrini)
belonging to family opisthorchiidae are parasites of the bile duct and gall
bladder of humans and fish-eating mammals.
The life cycle of these liver flukes is similar as well as their mode of
transmission. The mode of transmission is via ingestion of the metacercaria of
the parasite present in infected fish (2nd IH) that is eaten raw or undercooked.
a. Clonorchis sinensis
▪ Common name: Chinese Liver Fluke/Oriental
Liver Fluke
▪ Morphology:
o Adult:
➢ Flat, transparent, elongated,
rounded posteriorly and attenuated
anteriorly
➢ Cuticle – smooth and shiny
➢ Ventral sucker – smaller than oral
sucker Source:
https://www.cdc.gov/dpdx/clonorchiasis/im
ages/2/Clonorchis_new_labeled.jpg

➢ Testes – branched, arranged in tandem
o Ova:
➢ Yellowish-brown
with distinct convex
operculum
➢ Resembles an “old
fashion” electric
bulb
➢ Fully embryonated
Source:
when laid https://www.cdc.gov/dpdx/clonorchiasis/images/1/Clon
orchis_egg2.jpg
▪ Main Habitat:
o Bile passages/bile ducts of fish-eating mammals
▪ LIFE CYCLE:

1. The egg is fully mature when laid by the adult worm. It passes with
the bile to the intestine and escapes into the environment with the
feces.
2. The miracidium hatches only after the egg is ingested by the 1st
intermediate host (operculated snail)
3. Upon entry into the snail host, the miracidium transforms into a
sporocyst, which subsequently produces rediae
4. Each redia produces cercaria that are released into the surrounding
water
5. Upon contact with the second intermediate host (fresh water fish),
the cercaria attaches itself to the muscle or scale of the 2nd IH with its
suckers and encysts as metacercaria (infective stage)
6. Once a definitive host ingest a viable metacercaria, the
metacercaria excysts in the duodenum, and the young fluke moves
through the ampulla of Vater to the common bile duct, and then to
the distal biliary capillaries where it matures into an adult worm.
▪ 1st IH: Operculated snails
o Parafossarulus
o Bulimus
o Semisulcospira
o Alocinma
o Thiara
o Melanoides
▪ 2nd IH: Fresh water fish
o Tingea tingea
o Barbus barbus
o Clonorchiasis
o Carcinoma of the liver, adenocarcinoma of the gall bladder, as well
as pancreatitis have been associated with clonorchiasis
o Inflammation and intermittent obstruction of the biliary ducts →
irritation of the bile ducts

o Early infection:
➢ Leukocytosis and eosinophilia
o Acute infection:
➢ Chills and fever up to 40°C
➢ Enlargement and tenderness of the liver;
o Chronic infection:
➢ In the chronic stage, the clinical presentation may range from
asymptomatic to mild to severe disease
➢ Cirrhosis and portal hypertension are present
➢ Cholangitis, cholelithiasis, cholangiocarcinoma, and
pancreatitis
1. Eggs in biliary drainage (most practical diagnostic method)
2. Adult worms can be recovered through surgery
3. Liver scan
4. Phosphoglycerate kinase (a glycolytic enzyme) – can be used as an
immunoreagent in the serodiagnosis for clonorchiasis
5. Serologic tests: ELISA, EIA
▪ Treatment:
1. Praziquantel
2. Chloroquine phosphate
1. Treatment of positive cases with praziquantel in order to eliminate
human host reservoir
2. Health education for the promotion of cooked fish consumption in
order to prevent infection
3. Proper human waste disposal in order to interrupt the transmission of
the disease
b. Opistorchis felineus
▪ Common name: Cat Liver Fluke
▪ Distribution: Europe and Asia, including the former Soviet Union

▪ Morphology:
o Adult:
➢ Resembles Clonorchis sinensis
➢ Slightly shorter than C. sinensis
measuring about 8-12 mm
long and 1.5 to 3 mm wide
➢ Leaf-like in shape with
transparent tegument
➢ The main similarity between C.
sinensis and O. felineus is the
location of vitellaria, which are
found in the middle third of the
body at the level of the uterus
➢ Whereas the main difference is Source:
https://www.cdc.gov/dpdx/opisthorchia
in the morphology and sis/images/2/O_felineus_Korea.jpg
arrangement of testes
➢ Lobate testes, arranged obliquely
o Ova:
➢ Yellowish-brown, ovoid
➢ Measuring about 26-30 um by
15-17 um
➢ With distinct convex
operculum and a small
protuberance at the
abopercular end
➢ Fully embryonated when laid Source:
https://www.cdc.gov/dpdx/opisthorchiasis
▪ Main Habitat: /images/1/O_viverrini_egg_wtmt_BAM2.j
pg
o Distal ducts occasionally pancreatic
duct
▪ 1st snail IH: Bithynia leachi
▪ 2nd IH: Fresh water fish (Idus melanotus, Tinca tinca, Cyprinos carpio)

▪ LIFE CYCLE:
1. The adult flukes deposit fully developed eggs that are passed in the
feces
2. After ingestion by a suitable snail (first intermediate host) the eggs
will hatch releasing the miracidium
3. Once inside the snail host, the miracidia will undergo in several
developmental stages: mother and daughter sporocysts → redia →
cercaria

4. Cercariae are released from the snail into the surrounding water
seeking to its 2nd intermediate host
7. Upon contact with the second intermediate host (fresh water fish),
the cercaria attaches itself to the muscle or scale of the 2nd IH with its
suckers and encysts as metacercaria (infective stage)
5. After ingestion, the metacercariae excyst in the duodenum and
ascend through the ampulla of Vater into the biliary ducts, where
they attach and develop into adults
o Opistorchiasis
o Most infections are asymptomatic
o Mild cases:
➢ Dyspepsia, abdominal pain, diarrhea, constipation
o Acute infections:
➢ Resembles Katayama fever
➢ Fever, facial edema, lymphadenopathy, athralgia, rash,
eosinophilia
o Chronic infections:
➢ Hepatomegaly and malnutrition
o Rare cases:
➢ Cholangitis, cholecystitis, cholangiocarcinoma
1. Direct Fecal smear
2. Eggs in duodenal aspirate
▪ Treatment:
1. Praziquantel (drug of choice)

▪ Prevention and Control: Same as C. sinensis
c. Opistorchis viverrini
▪ Common name: Southeast Asian Liver
Fluke
▪ Distribution: Europe and Asia
▪ Morphology:
o Adult:
➢ Slightly similar as O. felineus
➢ Opistorchis viverrini is
differentiated from O. felineus Source:
https://www.cdc.gov/dpdx/opisthorchia
adult in that the testes of O. sis/images/2/O_viverrini_Korea.jpg
viverrini, which are positioned

close to each other and are more deeply lobulated
o Ova:
➢ Yellowish-brown, ovoid
➢ Measuring about 26-30
um by 15-17 um
➢ With distinct convex
operculum and a small
protuberance at the
abopercular end
➢ Fully embryonated when
Source:
laid https://www.cdc.gov/dpdx/opisthorchiasis/i
mages/1/O_viverrini_egg_wtmt_BAM1.jpg
▪ Main Habitat:
o Biliary ducts
▪ 1st snail IH: Bithynia spp.
▪ 2nd IH: Fresh water fish

▪ LIFE CYCLE:
1. The adult flukes deposit fully developed eggs that are passed in the
feces
2. After ingestion by a suitable snail (first intermediate host), the eggs
release miracidia which undergo in the snail several developmental
stages (sporocysts → rediae → cercariae)
3. Cercariae are released from the snail and is ready to penetrate
freshwater fish (second intermediate host), encysting as
metacercariae in the muscles or under the scales
4. The mammalian definitive host become infected by ingesting
undercooked fish containing metacercariae.

5. After ingestion, the metacercariae excyst in the duodenum and
ascend through the ampulla of Vater into the biliary ducts, where
they attach and develop into adults
o Opistorchiasis
o Most infections are asymptomatic
o Most pathologic manifestations result from inflammation and
intermittent obstruction of the biliary ducts.
o Clinical manifestations: same with O.felineus
▪ Laboratory diagnosis: Same as O. felineus
▪ Treatment: Same as O. felineus
▪ Prevention and Control: Same as O. felineus
*NOTE: the eggs of C. sinensis, O. viverrini, and O. felineus are difficult to differentiate.
However, when stained with KMnO4 and examined under a 400x magnification of a
light microscope show distinct melon-like ridges on the surface of Opistorchis viverrini
eggs.
d. Dicrocoelium dendriticum
▪ D. dendriticum is a common parasite of ruminants but humans can be
accidental definitive hosts
▪ Common name: Lancet fluke/Lanceolate
fluke
▪ Morphology:
o Adult:
➢ Slender, lancet-shaped, flat
➢ Testes: Slightly lobulated, situated
obliquely to each other anterior to
the small subglobous ovary
➢ Oral sucker: smaller than ventral
sucker Source:
https://i.pinimg.com/originals/44
➢ Vitellaria: Scanty /b1/92/44b192979f8e10afdb510
42adea45e6d.jpg

o Ova:
➢ Operculated and measure
35-45 µm long by 20-30 µm
wide
➢ The eggs are thick-shelled
and usually dark brown in
color
➢ Eggs are fully embryonated
Source:
when shed in feces. https://www.cdc.gov/dpdx/dicrocoelias
is/images/1/D_dendriticum_egg_wtmt
▪ Infective stage: METACERCARIA _JCG_C.jpg
▪ 1st snail IH: Land snails
o Cochicella
o Helicella
o Zebrina
▪ 2nd IH: Ants (Formica fusca)
▪ LIFE CYCLE:
1. Embryonated eggs containing miracidia are shed in feces of
definitive hosts, which are typically ruminants.
2. The eggs are then ingested by the first intermediate host (snail) and
then it will hatch into miracidia
3. They migrate through the gut wall and settle into the adjacent
vascular connective tissue, where they become mother sporocysts
4. The sporocysts migrate to the digestive gland where they give rise to
several daughter sporocysts. Inside each daughter sporocyst,
cercariae are produced
5. Cercariae migrate to the respiration chamber where they are shed
in slime ball from the snail image
6. After a slime ball is ingested by the second intermediate host (ant),
the cercariae will be released in the intestine and migrate to the
hemocoel where they become metacercariae
7. When the infected ant is eaten by a suitable definitive host, the
metacercariae excyst in the small intestine.

8. The worms migrate to the bile duct where they mature into adults
9. Humans can serve as definitive hosts after ingesting food
contaminated with infected ants

o Dricocoeliasis
o Chronic constipation and hepatomegaly
o Diagnosis is based on microscopic identification of eggs in the stool,
duodenal, and/or bile fluid
o Note that eggs may be detected in stool following consumption of
liver infected with adult flukes

o Additional specimens should be collected to distinguish this spurious
passage from a true infection.
o Adult flukes are rarely recovered
II. INTESTINAL FLUKES
A. Fasciolopsis buski
▪ Historical notes: It was first discovered by Busk in 1843, in the
duodenum of an East Indian sailor who died in
London. This fluke has been featured under a variety
of names, due to its size, different shapes, and
differences in integumentary spine pattern.
▪ Common name: Largest Intestinal Fluke
▪ Distribution: Asia and the Indian subcontinent,
especially in areas where humans raise pigs and
consume freshwater plants
▪ Morphology:
o Adult:
➢ Thick, flesh in color
➢ Cuticle is covered with transverse row of Source:
spines https://www.cdc.gov/dp
dx/fasciolopsiasis/image
➢ Testes: dendritic or highly branched and s/2/Fasciolopsis_buski_a
dult_GA.jpg
in tandem positioned at the posterior half of the worm
➢ Ovary: branched and located at the middle of the body
➢ Vitellaria: highly branched
o Ova:
➢ Large, has a clear thin shell
with a small operculum at
one end
➢ Resembles “hen’s egg”
➢ Immature when passed in
feces; usually it takes 3-7
Source:
weeks to embryonate in https://www.cdc.gov/dpdx/fasciolopsias
is/images/1/F_buski_egg_wtmt_BAM2.j
water
pg

▪ Main Habitat:
o Small intestine and sometimes in Large intestine and stomach
▪ 1st snail IH: Planorbid snails
o Segmentina
o Hippeutic
o Gyraulus
▪ 2nd IH: Aquatic plants
o Trapa bicornis (water caltrop)
o Eichuora (water hyacinth)
o Eliocharis tuberosa (water chestnut)
o Nymphaea lotus (lotus)
▪ Definitive host: Man and Pigs
▪ LIFE CYCLE:

1. Immature eggs are released together with the feces into the
water
2. The egg will embryonate in water which gives rise to miracidium in
3-7 weeks
3. The miracidium then seeks out and infects its first intermediate host
(snail)
4. Inside the snail, the miracidium transforms into a sporocyst which
subsequently produces mother rediae, mother rediae and finally,
cercaria
5. The cercariae leave the daughter rediae and undergo further
development in the snail tissues
6. Once the cercaria are released from the snail into the water,
cercariae attach themselves and encyst as metacercaria on the
surfaces of various aquatic plants (2nd IH)
7. Once a definitive host ingest aquatic plants encysted with
metacercaria (infective stage), the viable metacercaria excysts in
the duodenum and attaches to the intestinal wall where it
becomes sexually mature adult worms
o Fasciolopsiasis
o Most infections are light and asymptomatic
o In heavier infections, symptoms may include diarrhea, abdominal
pain, fever, ascites, anasarca, and intestinal obstruction
o The patient may experience generalized toxic and allergic
symptoms such as facial, abdominal walls, and lower limbs edema
o Profound intoxication can result into death
o Demonstration of eggs in the stool
o Vomitous sample may be used for detection of eggs
o NOTE: one should consider the resemblance between Fasciolopsis
buski and Fasciola eggs
▪ Treatment: Praziquantel
1. Since metacercaria are very sensitive to dryness, soaking of aquatic
plants in water should be avoided
2. The time between harvest and consumption could also be
prolonged to prevent infection

3. Washing of the plants to remove metacercaria or boiling them to kill
the parasite can also prevent infection
4. Swamps or ponds, where aquatic plants are cultivated should be
protected from pollution by untreated human or pig excreta
5. Community education
6. Eradication of snail intermediate hosts
7. Avoid eating raw aquatic plants
B. Family Echinostomatidae
The distinctive feature of echinostomes is a head collar bearing one or
two row of spines. The tegument usually bears spines or scales, especially
over the anterior part of the body. Echinostomes are parasites of birds and
mammals, and usually inhabits the intestine.
a. Echinostoma ilocanum
▪ Historical notes: Garrison first found the eggs of this fluke in native
prisoners in Manila in 1907, and later recovered 21 adult worms after
administration of oleoresin of aspidium. Tubangui in 1931 found that
the Norway rat was a reservoir of the infection.
▪ Common name: Garrison’s Intestinal fluke
▪ Distribution: Population of Ilocano in Luzon, Java and China where
dogs are infected
▪ Morphology:
o Adult:
➢ Elongated, reddish-gray
with horse-shoe shaped
collar of spines
surrounding the dorsal
and lateral sides of oral
sucker
➢ Cuticle: with minute
spine-like scales
Source:
https://stanford.edu/class/humbio
103/ParaSites2006/Echinostomiasi
s/Morphology.html

➢ Oral sucker is at the center of the body; ventral sucker is
located at the anterior fifth of the body a
➢ Testes: lobate; located posterior to ovary
➢ Ovary: globular; located anterior to the testes
➢ Uterus: looped, anterior the ovary
➢ Vitellaria: with small follicles which fills the lateral border of the
posterior two-thirds of the worm
o Ova:
➢ Straw-colored with
operculum at one end
➢ Measures 83-116 um by
58-69 um
➢ Immature when passed
in the feces
▪ Main Habitat: Source:
https://www.cdc.gov/dpdx/echinostomiasis/ima
o Small intestine ges/1/Echinostoma_egg_dog_BAM1.jpg
▪ 1st snail IH: Planorbid snails
o Gyraulus convexiusculus (Philippines and Java)
o Hippeutis (Philippines)
o Gyraulus prashadi (India)
▪ 2nd IH: Fresh water Mollusks
o Pila conica
Figure: An example of Pila conica, second IH of E. ilocanum

Source:
https://www.cdc.gov/dpdx/echinostomiasis/images/4/Gyr
aulus_Conchology_A.jpg

▪ LIFE CYCLE:
1. Unembryonated eggs are passed in feces of infected definitive hosts

and develop in water
2. Miracidia usually take about 3 weeks to mature before hatching.
3. After which, they swim freely and penetrate the first intermediate
host (snail)
4. The intramolluscan stages include a sporocyst, one or two
generations of rediae, and cercariae, which are released from the
snail.
5. The cercariae may encyst as metacercariae within the same first
intermediate host or leave the host and penetrate a new second
intermediate host
6. The definitive host becomes infected after eating metacercariae in
infected second intermediate hosts
7. Metacercariae excysts in the duodenum, and adults reside in the
small intestine (for some species, occasionally in the bile ducts or
large intestine)

o Inflammatory lesions may develop at the sites of attachment to
the intestinal wall together with a generalized toxic process
o Usual symptoms are intestinal colic and diarrhea
o Direct fecal smear examination
o Individuals can avoid infection by not eating raw snails in
endemic areas
C. Family Heterophyidae
The heterophyids are small or minute intestinal parasites of birds and
mammals. They are generally less than 1 mm long, pyriform or ovoid, with
spiny integument.
a. Heterophyes heterophyes
▪ Historical notes: this minute fluke was first found by Bilharz in 1851 at the
autopsy of a native Cairo.
▪ Common name:
o Dwarf Intestinal Fluke
o Von Seibold Fluke
▪ Morphology:
o Adult:
➢ It is an elongate, piriform worm,
with a broadly rounded posterior
end and a more attenuate
anterior end
➢ Cuticle is covered with minute
scale-like spine
➢ Testes: ovoid and placed side-
by-side located at the posterior
fifth of the body
➢ Ovary: globular located anterior
of the testes
➢ Vitellaria: with large polygonal
follicle in the lateral posterior
third of the body Source:
https://www.veterinaryparasit
o Ova:
ology.com/uploads/1/1/8/2/11
➢ Light brown in color, thick- 8230013/1b-heterophyes-
shelled, operculated minute egg heterophyes-logo_orig.jpg

➢ Contains fully developed miracidium
▪ Main Habitat: Small intestine
▪ 1st intermediate host: Brackish water snails
o Pirenella
o Cerithidea
▪ 2 intermediate host: Fish
nd
o Mugil cephalus
o Tilapia nilotica
LIFE CYCLE:
1. Mature eggs are passed out in the feces.

2. The miracidium hatches when the egg is ingested by the first snail
intermediate host (snail).
3. Inside the snail, the miracidium transforms into a sporocyst which eventually
develops into one or two generations of rediae that give rise to cercaria
4. Cercaria that are liberated from the snail encyst as metacercariae on or
under the scales, in the muscles, fins, tails, or gills of fish (2 nd IH)
5. Once the definitive host ingested a viable metacercaria, encysted in fish, the
metacercaria reaches the duodenum, it will undergo excystation, liberating a
young larva that attaches itself to the intestinal wall

6. The larva subsequently develops into a sexually mature adult worm that has a
typically short life span of less than one year
▪ Disease and Clinical manifestation:
o The main symptoms are mucoid diarrhea and colicky
abdominal pain
o Migration of the eggs to the heart, resulting in potentially fatal
myocardial and valvular damage
o Migration to other organs has also been reported
o In a study done in Compostela Valley, the most common
clinical manifestations observed were consistent with Peptic
Ulcer Disease or Acid Peptic Disease (APD)
1. Microscopic identification of eggs in the stool (NOTE: eggs are
indistinguishable from those of M. yokogawai, and resemble
those of Clonorchis and Opistorchis)
2. Definitive diagnosis is by detection of eggs in the stool using the
Kato-Katz method
▪ Treatment: Praziquantel (drug of choice)
➢ Preventive measures include avoiding ingestion of raw or
improperly cooked fish
b. Metagonimus yokogawai
▪ Common name:
o Yokogawa’s fluke
o Smallest human fluke
▪ Distribution: Mostly the Far East, Siberia, Balkan States, Israel, and Spain
▪ Morphology:
o Adult:
➢ 1 mm to 2.5 mm by
0.4 mm to 0.75 mm
in size
➢ Slightly resembles
H. heterophyes
➢ Cuticle: covered
with minute scale-
like spines
➢ Ventral sucker: Source:
large, situated at https://upload.wikimedia.org/wikipedia/commons
/a/ae/Metag_yokog_A.jpg
the right side of the
midline with its axis
in a diagonal line
o Ova:
➢ Indistinguishable from H. heterophyes
▪ 1ST intermediate host: Semisulcospira, Thiara, Hua

▪ 2nd intermediate host: Fresh water salmonoid fishes
▪ Disease: Metagonimiasis
➢ The main symptoms are diarrhea and colicky abdominal pain.
➢ Migration of the eggs to extraintestinal sites (heart, brain) can
occur, with resulting symptoms
➢ Microscopic identification of eggs in the stool
➢ However, the eggs are indistinguishable from those of H.
heterophyes and resemble those of Clonorchis and Opistorchis
➢ Specific diagnosis is based on identification of the adult fluke
evacuated after antihelminthic therapy, or found at autopsy
III. LUNG FLUKE

A. Paragonimus westermani
▪ Common name: Lung fluke, Oriental Lung Fluke

▪ Main Habitat: Lung Pockets
▪ Morphology:
o Ova:
➢ Operculated and ovoidal
(80-120 um)
➢ Yellowish-brown to dark or
golden brown on color
➢ Immature when laid with
a germ cell and many
yolk cells when oviposited
o Adults: Source:
https://www.cdc.gov/dpdx/parag
➢ Reddish brown in color onimiasis/images/1/Paragonimus
_egg_wtmt.jpg
➢ Tegument is covered with
single spaced spines
➢ Active state: spoon-shaped with one end contracted
and other end is elongated

➢ Preserved state: oval,
flattened, coffee bean-
shaped; has spinous cuticle
and suckers of equal size
➢ Male part – irregular lobed
testes are oblique to each
other and located at the
posterior third of the body
Source:
➢ Female part – ovary is lobed, https://www.cdc.gov/dpdx/par
agonimiasis/images/4/Parago
located anterior to the testes nimus_adult_hematoxylin2.jpg
on the right side opposite the

coiled uterus
➢ Vitellaria – at the extreme lateral fields on the entire
length of the body
▪ 1ST intermediate host: snail of genus Hua (Semisulcospira, Syncera and
Thiara, Pomatiopsis, Pomacea, Brotia asperata – most common in the
Philippines)
▪ 2nd intermediate host: Fresh water crabs or crayfish
▪ LIFE CYCLE:
1. The eggs are excreted unembryonated in the sputum, or alternately
they are swallowed and passed with stool
2. In the external environment, the eggs become embryonated, that
develops into miracidia
3. Once the miracidia hatch, it will seek for the first intermediate host,
a snail, and penetrate its soft tissues
4. Miracidia go through several developmental stages inside the snail:
sporocysts, rediae, with the latter giving rise to many cercariae
5. Cercaria will emerge from the snail
6. The cercariae invade the second intermediate host, a crustacean
such as a crab or crayfish, where they encyst and become
metacercariae.

7. This is the infective stage for the mammalian host
8. Human infection with P. westermani occurs by eating inadequately
cooked or pickled crab or crayfish that harbor metacercariae of
the parasite
9. The metacercariae excyst in the duodenum, penetrate through the
intestinal wall into the peritoneal cavity, then through the
abdominal wall and diaphragm into the lungs, where they become
encapsulated and develop into adults
10. The worms can also reach other organs and tissues, such as the
brain and striated muscles, respectively. However, when this takes
place completion of the life cycles is not achieved, because the
eggs laid cannot exit these sites.
11. Infections may persist for 20 years in humans.

▪ Disease: Paragonimiasis/Pulmonary Disturbances
o Clinical Manifestations:
➢ Patient with this disease most often complain of cough and
hemoptysis, clinical manifestations are consistent with
Pulmonary Tuberculosis (PTB). Hence, patients with this parasitic
infection are often misdiagnosed with PTB.
✓ Chronic bronchitis
✓ Hemoptysis
✓ Abdominal pain
✓ Dry cough and later produces blood stained sputum with
foul odor
✓ Chest pains, dyspnea
✓ Low grade fever, fatigue and generalized myalgia
▪ Laboratory Diagnosis:
o Definitive diagnosis is based on the detection of the characteristic
eggs in sputum, stool, or less frequently, in aspirated material from
abscessed or pleural effusion
o However, detection of eggs in sputum or feces of patients with
paragonimiasis is often very difficult
o X-ray – a typical finding is a ring-shadowed opacity, comprising
several contiguous cavities that give the appearance of a bunch
of grapes
o Serologic methods (CFT, EIA, Immunoblot assay)
▪ Treatment:
o Praziquantel
o Bithionol
o Thorough cooking of crab meat
o Health education to change food habits of the population

SYNTHESIS
Laboratory diagnosis of the flukes depends primarily on the careful microscopic

examination of stool, biliary drainage, duodenal drainage, or even sputum samples for
the presence of eggs. In addition to noting the specific specimen type, because select
organisms are found in certain samples, the organism’s size, shape, and features (e.g.
operculum, shoulders, and the presence and location of spines) aid in identification.
It is important to determine that the eggs if certain flukes, such as Fasciolopsis

and Fasciola, are indistinguishable and require further investigation for speciation.
Patient travel history, as well as clinical signs and symptoms and the possible recovery of
the adult fluke aid in the identification. The presence of shoulders helps to distinguish
Fasciola from Fasciolopsis.
Overall recovery of the adult flukes is considered to be a rare occurrence.

Nonetheless, it is important to have some idea of their appearance. With the exception
of the schistosomes, the monoecious flukes are basically flattened, leaf-shaped
organisms.
ASSESSMENT
SEATWORK: Complete the Table below. (15 POINTS)
Differences Between Monoecious and Dioecious Flukes

Monoecious Dioecious
1. Type and Number of
host
2. Infective stage
3. Appearance of the ova
4. Appearance of
cercariae
5. Appearance of the
Adult form
6. Main Habitat

QUIZ
form

REFERENCES
Inc.
Laboratory Identification of Parasites of Public Health Concern. Retrieved from

https://www.cdc.gov/globalhealth/ on July 27, 2020

LABORATORY
NAME: RATING:
Experiment No. 12
PHYLUM PLATYHELMINTHES - CLASS TREMATODA (MONOECIOUS FLUKES)
I. Background
Flukes are broadly oval/leaf-like in shape, bilaterally symmetrical along a long axis and are
usual1y flattened dorsoventrally. They vary in shape and size and the most characteristic
internal structures are the Acetabula or suckers. There are two suckers one surrounding the
oral cavity and the other on the ventral surface.
Digestive system consists of globular pharynx which extends from the mouth to a short
narrow esophagus. Intestines bifurcate into two straight or branching cecca that ends blindly.
Digestion is predominantly an extra-cellular process. Excretory system is bilaterally
symmetrical and open at posterior end with flame cells or solenocytes.
Nervous system is composed of a group of paired ganglion cells deposed like saddle on
the dorsum of pharynx or esophagus.
All are hermaphrodites/monoecious. Male genitalia consist of 2 testes: the vasa eferens
unites with the vas deferens that passes anteriorly into the seminal vesicle opening into the
common genital atrium. Female genitalia consist of a single ovary, oviduct, seminal receptacle,
vitelline glands & ootype. Mehli's gland and in some species, Lauren's canal. Some species (eg.
Heterophyids) have a genital sucker called the "gonotyl"
All trematodes lay eggs which upon embryonation give rise to ciliated embryo known as
miracidium. Eggs are provided with a lid called Operculum except for human blood flukes
which are non-operculated.
The trematodes need body of water to complete their life cycle. They require 2
intermediate hosts thus they are considered as heteroxenous.

1. To be able to know the morphology of the ova and adult Monoecious flukes
2. To describe the life cycle of Monoecious flukes
3. To know the mode of transmission Monoecious flukes
4. To be familiar with the diagnostic and infective stages of these flukes.
5. Identify these flukes using the distinguishing marks of their diagnostic stages

Microscope Prepared slides
Colored plates Reference Book

1. ova of Paragonimus westermani
2. ova of Clonorchis sinensis
3. ova of Fasciola hepatica
4. ova of Fasciolopsis buski
5. metacercariae
b. Print and Label: Diagrammatic
1. ova of Echinostoma ilocanum
2. ova of Heterophyid group
1. Opisthorcis felineus adult
2. Dicrocoelium dendriticum ova
ova of Paragonimus westermani ova of Clonorchis sinensis
ova of Fasciola hepatica ova of Fasciolopsis buski

Metacercariae
PRINT and LABEL (Diagnrammatic)
ova of Echinostoma ilocanum
ova of Heterophyid group

Opisthorcis felineus adult

1. __________________________
2. __________________________
3. __________________________
4. __________________________
5. __________________________
6. __________________________
7. __________________________
8. __________________________
9. __________________________
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwiE5KTs_ZjcAhVGHpQKHW5lAgoQjxx6BAgBE
AI&url=https%3A%2F%2Fweb.stanford.edu%2Fgroup%2Fparasites%2FParaSites2003%2FOpisthorchiasis%2Fmorph.htm&psig=AOvVaw
1Bgf2XIRFOqm-gE51iPbil&ust=1531464768597072)
Dicrocoelium dendriticum adult
1. ___________________________
2. ___________________________
3. ___________________________
4. ___________________________
5. ___________________________
6. ___________________________
7. ___________________________
8. ___________________________
9. ___________________________
10. ___________________________
11. ___________________________
12. ___________________________
13. ___________________________
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&ved=2ahUKEwiplPKeg5ncAhULKZQKHcTmAKsQjxx6BAgBEAI&ur
l=https%3A%2F%2Fwww.slideserve.com%2Ftuwa%2Fdicrocoelium-
dendriticum&psig=AOvVaw2iL5ZQUegKTv07gKkNXizn&ust=1531466150648725)

1. Describe the following

a) morphology of the ova and adult monoecious flukes
b) life cycle and mode of transmission
2. In a table form, differentiate the diagnostic & infective stage and distinguishing marks to identify
he flukes
3. In a table form, differentiate the different monoecious flukes based on common name, disease
caused, 1st intermediate host and 2nd intermediate host.

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF PROTOZOA
1. Identified each member of Protozoans according to morphology, and recognize

stage of the parasite being studied
confidentiality
LESSON PROPER
Protozoa are unicellular organisms and the lowest form of animal life. All
protozoa fall under Kingdom Protista. They have been divided into several phyla but the
major organisms causing disease in man belong to Phylum Sarcomastigophora, Phylum
Ciliophora, Phylum Apicomplexa, and Phylum Microspora.
Generally, protozoan parasites are provided with nucleus/nuclei, cytoplasm, an
outer lining membrane, and organelles. Among these are locomotory apparatus, which
include cilia, flagella, and pseudopodia. Many of these protozoans require a wet
environment for feeding, locomotion, osmoregulation, and reproduction. They form
infective stages called cysts, which are relatively resistant to environmental changes
compared to the vegetative stages called trophozoites. The parasitic species are
capable of multiplying within the host and may be transmitted through a biologic
vector within which they can also multiply.
GLOSSARY:
▪ Axostyle – rodlike supporting structure in some flagellates that gives rigidity

▪ Blepharoplast –basal body origin of the flagella that supports the undulating
membrane in blood flagellates
▪ Chromatin – deep-staining DNA-containing portion of the protozoa nucleus
▪ Chromatoid bar/body – rod-shaped structure of condensed RNA material within
the cytoplasm of certain amoeba cyst
▪ Cilia – small, hair-like cytoplasmic projections from a cell organism

▪ Costa – thin, firm, rodlike structure running along the base of the undulating
membrane of some flagellates
▪ Cyst – immotile stage protected by a resistant cyst wall formed by the parasite; it
is the INFECTIVE STAGE
▪ Cytostome – cell mouth present in some flagellates
▪ Ectoplasm – gelatinous cytoplasmic material beneath the cell membrane
▪ Endosome – small mass of chromatin within the nucleus; also known as the
karyosome
▪ Encystation – process when a cyst is formed
o FACTORS AFFECTING ENCYSTATIONS:
1. Deficiency or overabundance of food
2. Excess of catabolic products of the organism or associated
bacteria
3. Marked change in pH
4. Depletion/excess supply of oxygen
5. Overpopulation
▪ Excystation – transformation from a cyst to a trophozoite after the cystic form has
been ingested or swallowed by a host
▪ Flagellum – a tail-like extension of the ectoplasm that provides locomotion
moving in a whiplike motion
▪ Kinetoplast – intensely staining rod- or disc-shaped or spherical extranuclear DNA
structure found in parasitic flagellates near the base of the flagellum
▪ Pseudopod – protoplasmic extension of the trophozoites of amoeba that allows
movement and food engulfment
▪ Trophozoite – delicate and fragile; feeding, reproducing, motile stage
▪ Undulating membrane – protoplasmic membrane with a flagellar rim extending
out like a fin along the outer edge of the body of certain protozoa
CLASSIFICATION OF PROTOZOANS:
I. Phylum Sarcomastigophora
❖ Subphylum Sarcodina (Amoeba) – move by means of cytoplasmic
protrusions called pseudopodia; reproduce asexually. Subphylum

Sarcodina includes the amoeba namely, Entamoeba, Endolimax,
Iodamoeba, Acanthamoeba, and Naegleria.
A. Species which colonize the large intestines
▪ Entamoeba histolytica
▪ Entamoeba coli
▪ Endolimax nana
▪ Iodamoeba butschlii
▪ Entamoeba dispar
▪ Entamoeba hartmanni
B. Species which inhabit the oral cavity
▪ Entamoeba gingivalis
C. Species which inhabit the brain
▪ Naegleria spp.
▪ Acanthamoeba spp.
❖ Subphylum Mastigophora (Flagellates) – move by means of flagella;
reproduce asexually. Members of the subphylum Mastigophora
includes the atrial and hemoflagellates namely, Giardia, Chilomastix,
Trichomonas, Dientamoeba, Trypanosoma, and Leishmania.
A. Species which inhabit the digestive tract
▪ Giardia lamblia
▪ Pentatrichomonas hominis
▪ Chilomastix mesnili
▪ Retortamonas intestinalis
▪ Dientamoeba fragilis
B. Species which inhabit the oral cavity
▪ Trichomonas tenax
C. Species which inhabit the genitalia
▪ Trichomonas vaginalis
D. Species which inhabit the circulatory system
▪ Trypanosoma spp.
▪ Leishmania spp.

II. Phylum Apicomplexa (previously referred to as Sporozoa)
✓ The members of Phylum Apicomplexa have an apical complex at the
anterior end which consists of polar rings, subpellicular tubules, conoid,
rhoptries, and micronemes; these structures are involved in the
penetration and invasion of target cells.
✓ All members are parasitic
✓ Very important groups of parasites fall under Class Sporozoa, namely,
Plasmodia, Babesia, Toxoplasma, Isospora, Cryptosporidium, and
Cyclospora
✓ These organisms have been reported practically from all organ systems of
both humans and animals, specifically in the gastrointestinal tract,
genitourinary tract, CNS, respiratory tract, reticuloendothelial system,
blood, and blood cells, eyes, skin, and even oral cavity.
✓ Members of the Phylum Apicomplexa has a complex life cycle with
alternating sexual and asexual generations.
❖ Class Sporozoa
❖ Subclass Coccidia
❖ Suborder Haemospororina
1. Plasmodium vivax
2. Plasmodium falciparum
3. Plasmodium malariae
4. Plasmodium ovale
❖ Suborder Elmeriorina
1. Cryptosporidium spp.
2. Isospora spp.
3. Sarcocystis spp.
4. Toxoplasma gondii
❖ Subclass Piroplasmasina
1. Suborder Babesiidae
1. Babesia spp.
III. PHYLUM CILIOPHORA – locomotion is accomplished by means of cilia
❖ Class Kinetofragminophorea

2. Subclass Vestibulliferia
1. Balantidium coli
IV. PHYLUM MICROSPORIDIA (formerly classified with Sporoza)
✓ Rarely cause disease in immunocompetent persons, but may do so with
greater frequency in immunosuppressed persons.
1. Enterocytozoon bineusi
2. Encephalitozoon spp.
3. Vittaforma cornea
4. Pleitosphora spp.
A. Physical Functions:
1. Single cell-like organisms
2. Movement: through organelles like cilia, flagella, or pseudopod for
locomotion
3. Nutrition: through absorption of liquid or by the ingestion of solid particles or
by both processes
4. Excretion: the cytoplasm contains contractile vacuoles for maintenance of
osmotic pressure and elimination of waste product
5. Reproduction:
o Asexual
➢ Amitotic – single division
➢ Mitotic – repeated by binary binary fission of nucleus producing 2
daughter cells
o Sexual or Syngamy
B. Basic Structures:
1. Nucleus
2. Cytoplasm/endoplasm
➢ Inner, finely granular cytoplasm
➢ For storage, synthesis, and digestion of food
3. Ectoplasm
➢ For locomotion, respiration, and discharge of metabolic wastes
➢ Consists of:
o Plasma membrane
− Semi-permeable membrane
− For intake and out take of food
o Contractile vacuoles - Regulation of osmotic pressure
o Structures for locomotion
o Cytostome - Cell mouth; located laterally at the anterior end
o Cytophage - Cell anus

IDENTIFICATION, DIAGNOSTIC FEATURES, PREVENTION & CONTROL OF THE PHYLUM
SARCOMASTIGOPHORA: SUBPHYLUM SARCODINA (AMOEBA)
1. Identified each member of the Protozoans of the subphylum Sarcodina

according to morphology, and recognize stage of the parasite being studied
confidentiality
LESSON PROPER
The most important feature that separates amoebas from the other groups of
unicellular Protozoa is the means by which they move. Amoebas are equipped with the
ability to extend their cytoplasm in the form of pseudopods, which allows them to move
within their environment. With one exception, there are two morphologic forms in the
amoebic life cycle – trophozoites, the form that feeds, multiplies, and possesses
pseudopods, and cysts – the nonfeeding stage that is characterized by a thick
protective cell wall designed to protect the parasite from the harsh outside
environment when deemed necessary. It is important to note here that the nuclear
characteristics of trophozoites are basically identical to those of their corresponding
cysts.
The life cycles of all the intestinal amoeba are similar. The most common means
whereby amoebas are transferred to humans is through ingestion of the infective cyst in
contaminated food or water. In most cases, trophozoites are easily destroyed by the
gastric juices of the stomach. Trophozoites are also susceptible to the environment
outside the host. Therefore, trophozoites are not usually transmitted to humans.
Excystation, is the morphologic conversion from the cyst form into the
trophozoite form, which occurs in the ileocecal area of the intestine.

Encystation, is the conversion of trophozoites into cyst form. It usually occurs in
the intestine when the environment becomes unacceptable for continued trophozoite
multiplication. Contrary to the trophozoites, cysts are equipped with a protective cell
wall. The cell wall allows cysts to remain viable in the outside environment for a long
period of time. The ingestion of the infective cysts completes the typical intestinal
amebic life cycle.
General morphologic characteristics of a typical amoeba:
✓ Relatively small
✓ Typically lack contractile vacuoles
✓ Multiply by binary fission
✓ Undergo encystation EXCEPT: E. gingivalis
LIFE CYCLE OF AMEOBA:
1. Cysts and trophozoites are passed in feces

2. Cysts are typically found in formed stool, whereas trophozoites are
typically found in diarrheal stool
3. Infection by Entamoeba histolytica occurs by ingestion of mature cysts in
fecally contaminated food, water, or hands.
4. Excystation occurs in the small intestine and trophozoites are released,
which migrate to the large intestine.
5. The trophozoites multiply by binary fission and produce cysts and both
stages are passed in the feces
6. Because of the protection conferred by their walls, the cysts can survive
days to weeks in the external environment and are responsible for
transmission.
7. Trophozoites passed in the stool are rapidly destroyed once outside the
body; and if ingested, would not survive exposure to the gastric
environment.
8. In many cases, the trophozoites remain confined to the intestinal lumen
(noninvasive infection) of individuals who are asymptomatic carriers,
passing cysts in their stool.

9. In some patients the trophozoites invade the intestinal mucosa (intestinal
disease), or through the bloodstream, extraintestinal sites such as the liver,
brain, and lungs (extraintestinal disease).
10. It has been established that the invasive and noninvasive forms represent
two separate species, respectively E. histolytica and E. dispar.
11. These two species are morphologically indistinguishable unless E.
histolytica is observed with ingested red blood cells (erythrophagocystosis)
A. Entamoeba histolytica (By Schaudin)

➢ Most pathogenic amoeba in man
➢ It is the only amoeba that has the potential of tissue invasion

➢ Synonyms:
o Endamoeba histolytica
o Amoeba dysenteriae
o Entamoeba dysenteriae/Endamoeba dysenteriae
o Entamoeba tetragena
▪ Main Habitat: Large intestine of the host and other organs like liver, lungs, and
the brain
▪ Stages of Development:
A. Trophozoite
B. Precystic stage
➢ Transitional stage prior to the formation of cysts
➢ Colorless, round or oval, smaller than trophozoite but bigger than
cyst
➢ Devoid of food inclusion and movement is sluggish with no
progressive movement
C. Cystic stage
▪ Morphology of E. histolytica:
Trophozoites Cysts
Size 5-12 mcirons 3-10 microns

12-60 u 10-16 u

Pseudopodia Finger-like and rapidly extruded Absent
Motility Active, progressive, and Non-motile
unidirectional motility
Shape; ▪ No definite shape ▪ Spherical with a definite
cytoplasm ▪ Ectoplasm is thick, wide, outer cyst wall
refractile, and clearly ▪ Mature cyst has 4 nuclei
differentiated from (quadrinucleate)
endoplasm which is finely ▪ Immature cyst has 1-2
granular that may contain nuclei
ingested red blood cell BUT
NO bacteria or foreign
material
▪ Karyosome is centrally
located in the nucleus
Unstained ▪ Motile or rounded ▪ Nuclei not visible
▪ Nucleus not visible ▪ Chromatoid bar –
▪ RBCs are visible refractile
▪ Glycogen are refractile in
young cysts
Iodine-stained Nucleus visible Nucleus visible; chromatoid
bodies seldomly seen;
glycogen in young cysts are
visible
Chromatoid Absent Rods with rounded ends
matter (cigar/sausage-shaped)
Others ▪ Thermal death point at
50°C
▪ Resistant to urine, grow best
at anaerobic condition or
under reduced oxygen
tension

▪ Optimum growth at 37°C at
pH 7.0
▪ Mode of Transmission
➢ Ingestion of contaminated food and water containing cysts
➢ Direct contact to infected and uninfected persons (food handlers and
housekeepers)
➢ Faulty installation of water supple
➢ Faulty sanitary disposal
➢ Venereal transmission (sexually transmitted through fecal-oral contact)
▪ Factors Conducive to Invasion by Amoeba
➢ Temperature fluctuation in the host
➢ Abnormal secretory function
➢ Irritant foods
➢ Inadequate diet
➢ Inflammatory processes
▪ Pathogenesis and Symptomatology:
➢ E. histolytica was named by Schaudinn in1903 because of its ability to lyse
human tissues
➢ Its invasive process is initiated when the trophozoite stage is able to
penetrate through the mucus layer covering the colonic epithelium
➢ Disease caused:
a) Amebiasis
b) Amebic hepatitis
c) Amebic colitis/dysentery – clinically presents as gradual onset of
abdominal pain and diarrhea with or without blood and mucus
present in the stool
d) Amebic liver abscess – the most common extra-intestinal form
of amebiasis
e) Ameboma – clinically presents as a mass-like lesion with
abdominal pain; rare to occur
➢ Incubation period: 4-5 days

1. Intestinal/Primary Amebiasis:
➢ Metacystic trophozoites invade the cecum and cecal mucosa
➢ The trophozoites penetrate the cecal mucosa and epithelia by lytic
digestion aided by amoeboid movement
➢ Trophozoites burrow deeper with tendency to spread laterally by
continuous lysis of cells until they reach the submucosa to form flask-
shaped ulcers
2. Extra-intestinal/secondary/metastatic Amebiasis:
➢ Trophozoites demonstrated in every soft organ of the body
➢ Trophozoites which reach the muscularis mucosa frequently erode and
enter the lymphatics or walls of mesenteric venules on the floor of the
ulcers to reach other organs
a) Hepatic abscess
b) Pulmonary abscess
c) Cerebral amebiasis
d) Splenic abscess
e) Cutaneous abscess
▪ Signs and Symptoms:
✓ Asymptomatic in light infections (“luminal amebiasis”)
✓ Abdominal tenderness
✓ Diarrhea which may progress to dysentery in which there is passage of
blood and mucus over a period of weeks
✓ Constipation may be interspersed with diarrhea
✓ Peritonitis
✓ Dehydration
▪ Differential diagnosis of Amebic Dysentery from Bacillary Dysentery
➢ Acute Amebic colitis should be differentiated from bacillary dysentery
caused by bacteria such as Shigella, Yersinia, Salmonella, and Escherichia
coli
➢ Although stools may be grossly bloody, fever and significant elevated
leukocyte count are less common in amebic colitis

➢ Amebic colitis must be ruled out before steroid therapy for inflammatory
bowel disease is started because of the risk of developing toxic
megacolon
▪ Laboratory diagnosis
➢ The standard method of parasitologic diagnosis is through microscopic
detection of the trophozoites and cysts in stool specimen
➢ The detection of E. histolytica trophozoites with ingested red blood cells is
diagnostic of amebiasis
➢ Ideally, a minimum of 3 stool specimens collected in different days should
be examined
1. Direct Fecal Smear Examination and Permanently stained
preparations (using trichrome stain)
✓ For detection of trophozoites - fresh stool specimens
(diarrheic/watery stool) should be examined within 30
minutes after collection
✓ Cyst stage – usually found in formed or semi-formed stool
specimens
✓ Using the DFS with saline solution alone = one can
observed the motility of the trophozoite
✓ Using saline + methylene blue = Entamoeba spp. will stain
blue, thus, differentiating them from white blood cells.
✓ Using saline + iodine = the nucleus and karyosome can be
observed so that E. histolytica can be differentiated from
the non-pathogenic species (E. hartmanni, E. coli, E. nana)
2. Concentration Techniques
✓ Concentration methods such as FECT and MIFC are more
sensitive than the DFS for detection of cysts.
3. Purges saline by cathartic
4. Sigmoidoscopy material
5. Stool culture methods

✓ Balamuth’s medium
✓ Rice egg saline
✓ Locke egg serum
✓ Robinson’s and Inoki medium – more sensitive than stool
microscopy but is not routinely available
6. Serological methods
✓ Complement fixation test
✓ Indirect Immunofluorescence Assay
✓ Gel diffusion
✓ ELISA
✓ Latex agglutination assay
✓ PCR – useful in differentiation of luminal infections (E. dispar)
from invasive amoebiasis (E. histolytica)
NOTES TO REMEMBER:
❖ ANTIBODY DETECTION: most useful in patients with extraintestinal disease (i.e.
amebic liver abscess) when organisms are not generally found on stool examination
❖ ANTIGEN DETECTION: useful as an adjunct to microscopic diagnosis in detecting
parasites and to distinguish between pathogenic and non-pathogenic infections
(between E. histolytica and E. dispar infections
❖ Differentiation of E. histolytica and E. dispar is not possible by microscopy. This can
only be done by PCR, ELISA, and isoenzyme analysis.
❖ ELISA-based for stool is now commercially available showing a sensitivity of 80% and
specificity of 99%
❖ The use of PCR is limited by the requirement of sophisticated equipment
- Belizario & De Leon, 2004
7. For extra-intestinal amebiasis:

✓ Aspiration; biopsy
✓ Molecular methods for differentiating between E. histolytica
and E. dispar
✓ Liver scan
✓ Examination of aspirate (liver abscess)

▪ Treatment
A. For Intestinal Amebiasis
1. Asymptomatic or mild intestinal amebiasis
✓ Entero viaform – 0.25 gram 4x a day for 10 days
✓ Milibin – 0.5 gram 3x a day for 8 days
❖ Either of the above drugs should be combined with chloroquine or
aralen to take care of tissue invaders
2. Acute Amebic Dysentery
✓ Previous regiment is recommended, but in addition, emetine
HCl is given for first 3 days
✓ Tinidazole – 40-50 mg/kg body weight but not exceeding 2,000
mg in single dose repeated on the second day
B. For extra-intestinal Amebiasis – localized in the liver with or without extension
to the lungs (drugs to be given should be amoebicidal)
1. Emetine HCl – 1mg/KBW but not exceeding 65 mg for 7-10 days; after
a rest period of 2 weeks, the course is repeated
2. Chloroquine or aralen – 4 tablets (250 mg salt) first dose; 2 tablets daily
for 2 to 3 weeks
3. Tinidazole as in acute amebic dysentery
▪ Prevention and Control
1. Proper treatment of drinking water through filtration process and boiling of
water
2. Proper disposal and treatment of human excreta
3. Proper installation and maintenance of potable water
4. Proper processing and safe handling and preparation of food
5. Health education and promotion (e.g. Practice of handwashing, proper
use of latrines)
6. Use of iodine tablets to kill cysts
7. Uncooked vegetables should be scalded at 80’C for at least 30 seconds
B. Non-Pathogenic Amoeba of Man (Commensal Amoeba)
➢ The presence of commensal amoeba in the stool of an individual is
significant for two reasons:

1. They may be mistaken for the pathogenic E. histolytica
2. They are an indication of fecal contamination of food or water
➢ Accurate diagnosis of the commensal amoeba, is therefore, crucial.
LIFE CYCLE OF COMMENSAL AMOEBA:
1. Entamoeba coli, E. hartmanni, E. polecki, Endolimax nana, and

Iodamoeba buetschlii are generally considered nonpathogenic and
reside in the large intestine of the human host

2. Both cysts and trophozoites of these species are passed in stool and
considered diagnostic
3. Cysts are typically found in formed stool, whereas trophozoites are
typically found in diarrheal stool.
4. Colonization of the nonpathogenic amebae occurs after ingestion of
mature cysts in fecally-contaminated food, water, or fomites
5. Excystation occurs in the small intestine and trophozoites are released,
which migrate to the large intestine.
6. The trophozoites multiply by binary fission and produce cysts, and both
stages are passed in the feces
7. Because of the protection conferred by their cell walls, the cysts can
survive days to weeks in the external environment and are responsible for
transmission.
8. Trophozoites passed in the stool are rapidly destroyed once outside the
body, and if ingested would not survive exposure to the gastric
environment.
a. Entamoeba hartmanni (Von Prowasek, 1912)
▪ Common name: “small race E.
histolytica”
▪ Synonyms: Entamoeba minuta,
Entamoeba minutussima
▪ Habitat: Intestinal tract of man
▪ Morphology
E. hartmanni trophozoite
➢ Resembles E. histolytica except in
size (it is much smaller, and it does
not ingest red blood cells)
➢ Resembles E. nana in size
➢ Trophozoite: 4-12 u
➢ Cystic stage:
✓ Mature cyst: 5-10 um with 4 E. hartmanni cyst
nuclei (quadrinucleated)
✓ Immature cyst: usually have chromatoidal bars
➢ Chromatoidal bar: rice grain in shape
➢ The nucleus and location of karyosome resembles E. coli
➢ Motility: Sluggish
▪ Differences from E. histolytica:
✓ Trophozoite does not ingest red blood cells
✓ Motility is sluggish
✓ Chromatoidal bodies: shorter with tapered ends often referred
to as “rice grain shaped” or “thin fan like”
✓ Non-pathogenic or utmost causes only mild symptoms of
enteritis
b. Entamoeba coli (Graasi 1879; Casagrandi & Barbagalio 1895)
▪ Synonyms: Amoeba coli, Endamoeba hominis, Laschia coli
▪ Main habitat: large intestine of man
▪ Morphology:
o Nucleus: coarse, irregular peripheral chromatin; eccentric
karyosome
o Trophozoite:
✓ 15-20 um
✓ Coarse cytoplasm with many
vacuoles and ingested
bacteria
✓ Numerous nuclei (6-8 nuclei);
however nucleus is not easily
visualized E. coli trophozoite
✓ Ectoplasm is granular and not easily
differentiated from densely granular
✓ Short, blunt, and multiple pseudopods
o Cyst:
✓ Round or spherical
✓ 10-35 um
✓ 1-8 nuclei E. coli cyst

✓ Occasional chromatoidal bodies with splintered ends
(splinter-like/filamentous/thread-like with granules/whisk-
broom appearance)
✓ May have glycogen vacuole
▪ Motility: Sluggish; non-directional
▪ Mode of Transmission: Hand-to-mouth
▪ Laboratory diagnosis: Demonstration of cyst and trophozoite (DFS)

▪ Host-Parasite Interaction: non-pathogenic, no symptoms, no need for
treatment
▪ Prevention: personal hygiene and proper sanitary disposal of wastes
Differences Between E. histolytica and E. coli
Points of Differentiation Entamoeba histolytica Entamoeba coli
(Pathogenic) (Non-Pathogenic)
Trophozoite
Pseudopodia Finger-like Blunt and broader
Motility Progressive and Sluggish, non-progressive,
directional non-directional
Nucleus Bull’s eye nucleus Eccentric karyosome
Cytoplasm Clean-looking, with Dirty-looking, heavily
ingested RBCs vacuolated due to
ingested bacteria and
food particles
Cyst
Size Small race (non- Generally larger
pathogenic)
Large race (pathogenic)
Nuclear membrane Thin Thick
Number of nuclei 1-4 (infective) 1-8 nuclei
Chromatoidal bodies Sausage-shaped Splinter/broomstick
appearance

c. Endolimax nana (Wenyon & O’Connor, 1917; Brug 1918)
▪ Smallest intestinal protozoan of man
▪ Synonyms:
✓ Cross-eyed cyst
✓ Entamoeba nana
✓ Endolimax intestinalis
▪ Main Habitat: Cecum
▪ Laboratory Diagnosi: Direct Fecal Smear
▪ Motility: Sluggish, non-progressive
▪ Morphology:
o Nucleus:
✓ No peripheral cromatin; large irregular karyosome
✓ Karyosome appears as a large round dot
✓ Spherical or subspherical
o Trophozoite:
✓ 2-12 um
✓ Uninucleated
✓ Finely granular, vacuolated
cytoplasm (with narrow rim of
ectoplasm)
✓ Short pseudopod
o Cyst:
✓ 5-10 um
✓ Round to oval; usually oval
✓ 1-4 nucleus; possesses 4
nuclei when mature
✓
✓ Chromatoidal bodies are comma-shaped
▪ Laboratory Diagnosis: Direct Fecal Smear

d. Iodamoeba butschlii (Von Prowasek, 1911; Dobell, 1911)
▪ Common name: Iodine cyst
▪ Synonyms: Iodamoeba williamsi, Entamoeba williamsi, Entamoeba
butschlii, Endolimax williamsi,
▪ Main Habitat: Large intestine of man and swine
▪ Motility: Sluggishly-Progressively motility
▪ Morphology:
o Nucleus
✓ Spherical
✓ No peripheral chromatin
✓ Large karyosome surrounded by layer of small granules
✓ The karyosome is large, irregular, and rounded with a
cluster of achromatic granules
o Trophozoite
✓ 8-20 um
✓ Coarsely granular cytoplasm
with vacuoles and bacteria
✓ Blunt pseudopods (sluggishly-
progressively motility)
✓
✓ Small with fairly active, progressive movement
✓ Ectoplasm is clear
o Cyst
✓ 5-20 um
✓ Ovoid
✓ Only one nucleus when mature
✓ Prominent glycogen vacuole
(iodine-staining)
e. Entamoeba gingivalis (Gros, 1849; Brumpt, 1913)
▪ Synonyms: Amoeba gingivalis, Amoeba buccalis, Entamoeba buccalis

▪ Main Habitat: Found in the mouth,
chiefly in the tartar of the teeth and
gingival pocket
▪ Mode of Transmission:
✓ Kissing or droplet spray
✓ Contaminated drinking
utensils and dental utensils
▪ Morphology:
o NO CYSTIC STAGE: exists only as a trophozoite and does not
undergo encystation
o Trophozoite:
✓ Measures 5-35 um in diameter
✓ Extrudes pseudopodia, similar to E. histolytica but does
not exhibit progressive locomotion
✓
✓ Small and centrally located karyosome
f. Entamoeba dispar
▪ Morphologically identical to E. histolytica
▪ The only difference of E. dispar from E. histolytica is that it cannot ingest
red blood cells
▪ Serological testing may be useful to differentiate E. histolytica from E.
dispar
g. Entamoeba polecki
▪ Common parasite of pigs and monkeys
▪ Rarely to cause disease in humans
h. Dientamoeba fragilis (Wenyon, 1909; Dobelli, 1918)
▪ Originally described as amoeba, but is
actually a flagellate with only the
trophozoite stage known. It is now
classified among the Trichomonads
(despite the missing flagellum)
because of the following:

✓ Binucleated trophozoite
✓ Absence of the cyst stage
✓ Electron microscopic evidence of rudiments
✓ Resembles Trichomonads antigenically and ultrastructurally
▪ Pathogenic to man
▪ Main Habitat: mucosal crypts of the cecum
▪ Usually in co-infection with E. vermicularis
▪ Identification:
➢ Could be recognized only in fresh liquid or soft stool specimen
➢ Prompt fixation with PVA or Schaudinn’s is helpful
▪ Morphology:
✓ Small size, 2 nuclei
✓ Circular appearance at rest
✓ Rapid action of the multiple leaf-shaped pseudopods that gives
a stellate appearance, and explosive disintegration in water
C. Free Living Pathogenic Amoeba of Man
➢ Among the numerous free-living amoeba of soil and water habitats, certain
species belonging to two genera – Acanthamoeba and Naegleria – are
facultative parasites of man.
➢ Habitat: Stagnant water, brackish and ocean sediments, thermal pools,
swimming pools, polluted soil, sewage disposal systems
➢ Usually inhabits the Central Nervous System once it enters a human host
A. Naegleria species (N. fowleri)
▪ Mode of Transmission: can be acquired while diving and swimming
during hot weather in brackish or fresh water including swimming pools
▪ Morphology:
o Trophozoite
✓ Size: 10-35 um
✓ Trophozoite can assume
limax form or become
ameboflagellate
✓ Has both amoeba and flagellated form

❖ Ameboid: has a blunt pseudopodia and a vesicular
nucleus with a large karyosome and sparse granules
of peripheral chromatin
❖ Flagellated: elongated and bears two equal and
anteriorly located flagella
✓ NOTE: when inside the host, Naegleria trophozoites do not
exhibit the flagellated stage and cysts are not also formed;
only the ameboid form is present inside the host
o Cyst:
✓ Size: 7-10 um in diameter
✓ Round
✓ Cyst wall is smooth and double, with the outer wall
perforated by 3 – 8 pores (ostioles)
✓ Single nucleus
✓ Spherical chromatoid bodies
▪ Life Cycle:
1. The parasite enters the host through nasal passages (nose) while
swimming or diving in contaminated bodies of water particularly
during warm weather
2. From nasal passages, the trophozoites migrate along the olfactory
nerves through the cribriform plates and into the meninges and
cerebral hemispheres of the brain
3. Note that when inside the host, Naegleria trophozotes do not
exhibit the flagellated stage, and cysts are not also formed.

▪ Pathogenesis:
➢ Major causative agent of Primary Amebic Meningoencephalitis
(PAM)
➢ Primary Amebic Meningoencephalitis:
− Usually fatal within a week of onset

− On autopsy examination (of mice and animals), the normal
architecture of the brain particularly the olfactory lobes and
cerebral cortex is completely destroyed. (“Brain-eating
amoeba”)
− Causes purulent spinal fluid with motile amoeba
▪ Laboratory Diagnosis: Stained smears of culture material (demonstration
of the trophozoites in CSF)
B. Acanthamoeba spp. (A. castellani, A. culbertsoni, A. hutchetti, A. polyphaga)
✓ Swimming in
contaminated water
✓ Using inadequately
disinfected contact
lenses
▪ Portal of entry: broken or ulcerated skin or eye, lungs, genitourinary tract
▪ Morphology:
o Trophozoite
✓ 10-45 um
✓ Has single vesicular nucleus and also a large karyosome
o Cyst
✓ Uninucleated and double walled
✓ 16 um
▪ Life cycle:
1. Acanthamoeba has two stages; cysts image and trophozoites
image in its life cycle and lacks a flagellate stage.
2. The trophozoites are the infective forms, although both cysts and
trophozoites can enter the body through various means.
3. Entry can occur through the eye, the nasal passages, to the lower
respiratory tract, or ulcerated or broken skin
4. When Acanthamoeba spp. enters the eye it can cause severe
keratitis in healthy individuals, particularly contact lens users

5. When it enters the respiratory system or through the skin, it can
invade the central nervous system by hematogenous dissemination
causing granulomatous amebic encephalitis (GAE), or skin lesions
in individuals with compromised immune systems.
6. Both Acanthamoeba spp. cysts and trophozoites are found in the
tissue.
▪ Diseases:
o Ulcerative Acanthemoeba Keratitis in contact lens wearers
− Causes keratitis, acquired from trauma and contact lens
wear
− Characterized by severe ocular pain (invasion of cornea or
interior of the eye)
o Granulomatous Amebic Encephalitis

− Chronic central nervous system infection; generally in
debilitated or immunocompromised patients
o Chronic Granulomatous lesions in the brain, skin, kidneys, liver,
spleen, uterus, and prostate
o Microabscesses in the lungs and pancreas
o Stained smears of culture material
o Histologic examination of brain
o Trophozoites and cysts in corneal scrapings
▪ Treatment: Amphotericin B (given intravenously)
▪ Prevention and control:
✓ Avoid swimming in stagnant water or thermal water
✓ Salination of water up to 0.7%
SYNTHESIS
The species of amoeba that are commonly found in human fecal specimens
are E. histolytica, E. dispar, E. hartmanni, E. coli, E. polecki, E. nana, and I. butschlii. They
are mainly differentiated on the basis of structure and size, thus, proper determination
of the organism size is essential identifying the amoebas. The appearance of key
nuclear characteristics, such as the number of nuclei present and the positioning of the
nuclear structures is crucial to differentiate the amoebas correctly. The presence of
other amebic structures and characteristics such as cytoplasmic inclusions and motility
also aids in the identification of amoeba. It is important to note that only one of the
intestinal amoebas, E. histolytica, may produce characteristic symptoms, and is
universally considered to be a pathogen.

ASSESSMENT
SEATWORK: Complete the table below. Point of the differences necessary for the
differential diagnosis of amebic dysentery from bacillary dysentery. (20 POINTS)
Bacillary dysentery Amebic Dysentery

Onset Acute Gradual
Signs and Symptoms
Stool analysis findings
Odor of the stool
Microscopic findings in stool
Prognosis
QUIZ
form

REFERENCES
Inc.
Laboratory Identification of Parasites of Public Health Concern. Retrieved from

https://www.cdc.gov/globalhealth/ on July 27, 2020

LABORATORY
NAME: RATING:
Experiment No. 13
PHYLUM SARCOMASTlGOPHORA - SUBPHYLUM SARCODINA
I. Background
The amoebae fall under Superclass Rhizopoda. They move by means of lobe-like structures
projecting from the ectoplasm of the organism referred to as pseudopodia. They multiply by binary
fission, and in general, they exhibit the following stages of development: trophozoites, pre-cysts,
cysts, and metacystic trophozoites.
Several species of intestinal amoebae are known to infect humans. Among these are the
following:
1. Entamoeba histolvtica
2. Entamoeba coli
3. Entamoeba hartmanni
4. Entamoeba dispar
5. Endolimax nana
6. Iodamoeba butschlii
The taxonomic grouping of the amoebae is mainly based in the nuclear structural
characteristics for the genus. Entamoeba species have a delicate karyosome also known as
nucleolus at the center or near the center of the nucleus. The bounding nuclear membrane is lined
with chromatin granules. Endolimax species are provided with fairly large karyosome with
chromatin fibrils, which may or may not be anchored to the nuclear membrane. While genus
Iodamoeba has a large karyosome at the center of the nucleus surrounded with achromatic
granules.
Both cyst and trophozoites are important in the identification of the amoebae hence basically,
there is a need to use a calibrated microscope. Laboratory diagnosticians also look into the
structures observed in the cytoplasm of the organism like ingested red cells, appearance of
chromatoidal bars, presence of glycogen mass, food vacuoles and even bacteria.

1. To be able to know the morphology of amoeba
2. To describe the life cycle of amoeba
3. To know the mode of transmission amoeba
4. To be familiar with the diagnostic and infective stages of these amoebae
5. Identify the amoebae using the distinguishing marks of their diagnostic stages

Microscope
Prepared slides
Colored plates
Reference Book
1. cyst and trophozoite of Entamoeba histolytica
2. cyst and trophozoite of Entamoeba coli
b. Print and Label: Diagrammatic (Cyst and trophozoite)
1. Entamoeba hartmanni
2. Endolimax nana
3. Iodamoeba butschlii
4. Dientamoeba fragilis trophozoite
5. Entamoeba gingivalis trophozoite
cyst of Entamoeba histolytica trophozoite of Entamoeba histolytica
cyst of Entamoeba coli trophozoite of Entamoeba coli

PRINT AND LABEL (Diagrammatic)
PROTOZOA CYST TROPHOZOITE
Entamoeba hartmanni
Endolimax nana
Iodamoeba butschlii
Dientamoeba fragilis trophozoite
Entamoeba gingivalis trophozoite

Questions for enrichment (answers on the space provided)
1. Describe the general morphology of amoeba

2. Describe the life cycle and MOT of amoeba
3. In a table form, differentiate the different types of amoeba based on diagnostic stage and
infective stage.
4. In a table form, differentiate cyst from trophozoite
5. Briefly describe and give the laboratory diagnosis of the following:
a) E. histolytica
b) E. coli
c) E. hartmanni
d) E. nana
e) D. fragilis
f) E. gingivalis
g) I. butschlii

SARCOMASTIGOPHORA: SUBPHYLUM MASTIGOPHORA (INTESTINAL FLAGELLATES)
1. Identified each member of the Protozoans of the subphylum Masitogophora:

Intestinal Flagellates according to morphology, and recognize stage of the
parasite being studied
confidentiality
LESSON PROPER
In this section of the module consists of a discussion in the morphologic features,

laboratory diagnosis, life cycle, epidemiology, clinical symptoms, treatment, and
prevention and control of the eight members of the flagellates, each of which is known
to infect humans.
The flagellates belong to the phylum Protozoa and are members of the
subphylum Mastigophora. The flagellates can be categorized into two groups, intestinal
and atrial. Members of this class lack chromatophores and thus depend on previously
manufactured plant and animal foods. Their nutrition is holozoic or parasitic, and their
form may be simple or quite complex, but as a rule, they have a single nucleus and a
neuromotor apparatus. The neuromotor consists of a blepharoplast, which constitutes
the kinetoplast (energizing component), and an axoneme, with or without a free
flagellum. The axial structure of the flagellum is a continuation of the axoneme, which is
composed of one or more fibrils. Some species of flagellate protozoa have a
rudimentary mouth, the cytostome. Reproduction is essentially asexual and is by
longitudinal binary fission. In majority of the family groups, the life cycle is simple, but in
trypanosomes, the organisms are dimorphic or polymorphic.
o Most are free-living or parasitic
o Possess whip-like locomotory organelle called flagella
o Motor component: flagella and axonemes
o Neuromotor apparatus: kinetoplast which contains of blepharoplast and
parabasal body
o Mode of Reproduction: longitudinal binary fission

I. INTESTINAL FLAGELLATES
A. Giardia lamblia
▪ The only pathogenic intestinal flagellates
found only in man
▪ Synonyms:
➢
➢ Giardia intestinalis, Giardia
duodenalis, Cercomonas intestinalis,
Megastoma enterica, Lamblia
intestinalis, Giardia enterica
➢ Ingestion of cyst usually through
contaminated water
➢ Oral-anal route
▪ Main Habitat: Duodenal area of small intestine and
gall bladder
▪ Morphology:
o
o Trophozoite: (Invasive stage)
✓ Pear-shaped/Pyriform
✓ Rounded anteriorly and pointed posteriorly
✓ Resembles “old man in eye glasses” or “tennis racket”
appearance
✓ Bilaterally symmetrical: two media
bodies
✓ Measures 9.5 – 21 um by 5 – 10 um
✓ With large sucking discs on the ventral,
concave side and convex on the
dorsal side, occupying about ¾ of the
flat ventral surface
✓ Presence of 2 nuclei with large, central
karyosome
✓ 2 axostyles, 2 blepharoplast, 2 deeply
staining bars
✓ 4 pairs of flagella: one pair of laterally crossed flagella,
one pair of central flagella, a lateral pair of uncrossed
flagella, and one pair of posterior flagella
❖ Motility: jerky falling leaf/kite-like/spinning/flip-flop
motility

o Cyst: (Infective stage)
✓ Ovoid/ellipsoidal;
measures 8 – 12 um
by 6 – 10 um .
✓ 2 – 4 nuclei at
anterior end
✓ Thick double wall
(“double-walled
cyst”); cytoplasm
shrinks away from
the cell wall
✓ Axostyle and fibrillar remnants of locomotory apparatus
present
▪ Life Cycle:

1. Ingestion of viable cysts from contaminated food and water
2. Excystation occurs in the duodenum and become trophozoites
3. Trophozoite inhabits the mucosa of duodenum and proximal
jejunum (optimal pH: 6.4 – 7.0)
4. Reproduce by binary fission
5. Encystation occurs in the large intestines with water reabsorption
▪ Disease: Giardiasis or Flagellate diarrhea (also known as Traveller’s

diarrhea)
▪ Clinical Manifestations:
➢ Majority of persons infected are asymptomatic or is manifested
as a self-limiting acute onset diarrhea, usually associated with
nausea, anorexia, and crampy abdominal pain
➢ Common among pre-school children and immunocompromised
patients (AIDS patients)
a. Duodenal Involvement (Duodenitis)
✓ Irritation with excess secretion of mucus and
dehydration
✓ dull epigastric pain
✓ chronic diarrhea with steatorrheic stool containing
increased amounts of fat and mucus but no blood
b. Gall bladder involvement (Cholangitis)
✓ Associated with gallbladder colic and jaundice
due to obstruction of the bile passages
✓ Irritation or edema of the ampulla of Vater
1. DFS: stained or unstained
2. ZnSO4 flotation technique – for cyst concentration
3. Duodenal aspiration (Entero test)
4. Fluoroscopy – may demonstrate hypermotility at the duodenal
and jejunal levels
5. X-ray to reveal mucosal defects
6. Culture method
▪ Treatment:
➢ Metronidazole – drug of choice
1. Personal and community hygiene
2. Sanitation and cleanliness
B. Chilomastix mesnili
▪ Nonpathogenic
▪ Synonym: Shepherd’s crook, Chilomastix hominis
▪ Distribution: Harmless commensal which is worldwide in distribution
▪ More prevalent in warm than cooler climate
▪ Main Habitat: cecal region of the large intestine
▪ Mode of Transmission: Ingestion of cyst
▪ Morphology:
o Trophozoite:
✓ Asymmetrical pear-
shaped (due to the
cytostome) measuring
about 6 – 20 um
✓ Broad anterior,
tapering toward the
posterior end
✓ Spiral groove
extending through the
middle portion of the
body
✓ 3 pairs of flagella and a more delicate one within the
prominent cytostome
✓ Cytoplasm is delicately granular with numerous food
vacuoles
✓ 1 nucleus with central karyosome
✓ Motility: Boring or spiral forward movement, corkscrew,
clockwise, twisting motility
o .Cyst
✓ 7 – 10 um; thick walled
✓ Pear- or lemon-shaped,
rounded at one end and
conical at the other end
with knob-like
protuberance projection
✓ 1 spherical nucleus with
central karyosome
▪ Prevention: Proper hygiene and sanitation
C. Enteromonas hominis
▪ Morphology:
o Trophozoite
✓ Pear-shaped or ovoid
✓ No cytostome
✓ 4 flagella: 3 anterior and 1 which closely adheres to the
flattened surface of one side and trails off posteriorly as a
free flagellum
✓ Motility: Jerky motility
o Cyst: pear-shaped, uninucleated
D. Embadomonas intestinalis
▪ Morphology:
o Trophozoite
✓ Measures 4 – 9 um by 3 – 10 um
✓ Characteristic cleft – like cytostome may be seen near
the nucleus
✓ 2 anterior flagella
o Cyst: pear-shaped, uninucleated
II. ATRIAL FLAGELLATES
Genus Trichomonas
▪ Exist only as trophozoites; NO CYSTIC STAGE
▪ Trophozoite stage is the infective stage and pathogenic stage
▪ Multiplication is through longitudinal binary fission
▪ Motility: Fast jerky tumbling movement
▪ Morphology:
➢ Nucleus: single, spherical or subspherical in the mid-line near the
anterior pole
➢ Cytoplasm: highly vacuolated
➢ Rounded anterior and pointed posterior end (Pyriform in shape)
with:
o Axostyle – rod-like structure arising from or near the anterior
end and extends through the entire body, protruding as a
rather sharp spike through the posterior end of the
cytoplasm; functions for anchorage
o Cytostome – small, located on one side of the anterior end
o Blepharoplast – located between the nucleus and the
anterior margin of the organism
o Flagella – 4 free flagella and 1 which runs along the
undulating membrane
Trichomonas vaginalis
▪ Optimum pH for survival: 5.2 to 6.4
▪ In the healthy female, the normally acid vaginal secretions of pH 3.8 – 4.4
deter its survival
▪ Pathogenesis and Symptomatology:
1. Inflammation of the vaginal mucosa occurs several days after
inoculation of the trophozoites, after which, polymorphonuclears
become numerous and epithelial cells desquamate
➢ Vaginal secretions are liquid in character, greenish yellow in
color and very irritating that it causes intense itchiness or
burning sensation
➢ Strawberry cervix in appearance
2. In most cases, T. vaginalis infection in females present as a
symptomatic vaginitis, although chronic infection may be
asymptomatic
3. In males – trichomoniasis is latent or symptomless during the acute
stage and it usually becomes chronic urethritis
▪ Treatment:
1. Oral metronidazole – 250 mg 3x a day for 7 days results in 90-98%
cure; for better patient compliance, oral metronidazole 2 grams
single dose gas 86% cure rate
2. Acid douche (pH of 3.0); Use prophylactic devices

Life Cycle:
1. Trichomonas vaginalis resides in the female lower genital tract and the
male urethra and prostate, where it replicates by binary fission
2. The parasite does not appear to have a cyst form, and does not
survive well in the external environment.
3. Trichomonas vaginalis is transmitted among humans, its only known
host, primarily by sexual intercourse

Trichomonas species
Differences T. tenax T. hominis T. vaginalis
(Dobell, 1939) (Leukart, 1879) (Donne, 1837)
Habitat Buccal cavity, Cecum Vagina, prostate
tartar of the teeth gland
Size Smallest Medium Largest
Nucleus Round Ovoidal Ovoidal
Undulating 2/3 of the body As long as the Less than ½ of the
membrane length body length body length
Inclusion bodies NONE With siderophil
bodies in the
cytoplasm
Flagella 4 anterior and 1 posterior
Cytostome Inconspicuous Highly Very conspicuous
conspicuous
Pathogenicity NON-PATHOGENIC Vaginitis,

prostatitis,
urethritis, itching
and irritation,
burning sensation
of urine
Mode of Droplet spray Ingestion of Sexual contact
Transmission Kissing contaminated and rarely
Use of food and drink congenital
contaminated
dishes or drinking
glass
Specimen of Oral scraping Stool Vaginal swab,
choice urine, urethral
discharge
Prevention Oral hygiene Sanitation and Avoid
personal hygiene promiscuous
sexual intercourse

SYNTHESIS
The parasitic flagellate Protozoa is earlier classifications were placed in the class
or superclass Mastigophora together with all species of one-celled animals that bear
one to several long, delicate, thread-like extensions of the cytoplasm, which are termed
flagella. Some species of flagellate protozoa have a rudimentary mouth, called the
cytostome. Reproduction is through longitudinal binary fission. The only intestinal
flagellate to cause disease in humans is G. lamblia, while among the trichomonas
species, the only specie known to be pathogenic in man is T. vaginalis.
ASSESSMENT
QUIZ
form

REFERENCES
Inc.

SECTION 5C: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
SARCOMASTIGOPHORA: SUBPHYLUM MASTIGOPHORA: BLOOD AND TISSUE FLAGELLATES
1. Identified each member of the Protozoans of the subphylum Masitogophora:

Blood and Tissue Flagellates according to morphology, and recognize stage of
the parasite being studied
confidentiality
LESSON PROPER
The flagellate protozoa that live in the blood and tissues of human host all
belong to the order Kinetoplastida, family Trypanosomatidae. This family includes
species that have a single flagellum, a nucleus, and a kinetoplast from which the
flagellum arises. The kinetoplast consists of a deeply stained parabasal body and an
adjacent dot-like blepharoplast. The blepharoplast and parabasal body are
connected by one or possibly more delicate fibrils. The inner portion of the flagellum
extending from the blepharoplast to the surface of the body is axoneme, or axial
filament.
Species of this family may exist in two or more of four forms or states, which are
named after genera exemplifying these forms: Leptomonas, Crithidia, and
Trypanosoma.
GENERAL CHARACTERISITICS:
▪ Include the Leishmaniasis and trypanosomes
▪ May infect the blood, lymph nodes, muscles, and reticuloendothelial system
▪ Multiply in the blood (hemoflagellate) and tissue of humans
▪ Basic Structures:
1. Single flagellum, a nucleus, and a kinetoplast
2. Blepharoplast and the parabasal body
3. Axial filament/axoneme
▪ All species require an arthropod intermediate host
▪ Four stages:
1. Amastigote (Leishman-Donovan body) or leishmanial form
2. Promastigote or leptomonal form
3. Epimastigote or crithidial form
4. Trypomastigote or trypanosomal form

Trypanosoma and Leishmania: Four stages
Stage Description Common Found in
name
Amastigote No flagella Leishmanial L. donovani
form L. tropica
L. braziliensis
T. cruzi
Promastigote Flagella in front of the nucleus Leptomonad Sand fly

form (Leishmania vector)
Trypanosoma cruzi
(transitional phase)
Epimastigote Flagella in the center of organism Crithidial form Tse tse fly
Reduvid bug
(trypanosome
vector)
Trypomastigote Flagella originates at posterior end Trypanosomal T. brucei

of the organism form rhodesiense
T. brucei
gambiense
T. cruzi

Genus Leishmania
1. Leishmaniasis is transmitted by the bite of infected female phlebotomine

sand flies.
2. The sand flies inject the infective stage (promastigotes) from their
proboscis during blood meals
3. Promastigotes that reach the puncture wound are phagocytized by
macrophages and other types of mononuclear phagocytic cells.
4. Promastigotes transform in these cells into the tissue stage of the parasite
(amastigotes, which multiply by simple division and proceed to infect
other mononuclear phagocytic cells)
5. Sand flies become infected by ingesting (amastigote) infected cells
during blood meals
6. In sand flies, amastigotes transform into promastigotes, develop in the
gut, and migrate to the proboscis

Leismania species
Features Leishmania tropica Leishmania braziliensis Leishmania donovani
Clinical disease Old world American or Visceral leishmaniasis,

cutaneous Mucocutaneous Kala-azar fever, Dum-
leishmaniasis, leishmaniasis, Espundia, dum fever, Death
locally known as Uta, Chiclero ulcer, fever, tropical
Oriental sore, Delhi Nasopharyngeal splenomegaly
boil, Aleppo leishmaniasis, New world
button, Forest yaws cutaneous leishmaniasis
Baghdad or Jericho
boil
Habitat Endothelial cells of Mucocutaneous Endothelial cells of the
the infected skin junctions, particularly the reticuloendothelial
capillaries and nasal septum, mouth, system: liver, spleen,
within the and pharynx bone marrow, and
cytoplasm of large visceral lymph nodes
phagocytic and fixed tissue
monocytes macrophages
Vector Phlebotomus Phlebotomus peruensis Phlebotomus
papatasii Phlebotomus verrucarum argentipes
Phlebotomus
sergenti
Mode of Bite of sand fly Bite of sand fly Bite of sand fly
Transmission (Phlebotomus spp.) (Phlebotomus spp.) (Phlebotomus spp.)
Infective stage Promastigotes Promastigotes (released Promastigotes
(released via bite via bite of a sand fly) (released via bite of a
of a sand fly) sand fly)
Pathology Localized Primary lesion is similar to Splenomegaly
cutaneous that of Leishmania associated with
infection which tropica except the ulcer severe anemia
gives rise to a produced is a weeping
macule, then a lesion without a granular
papule, a raised base
lesion with
depressed
ulcerated center
Clinical ▪ Presence of ▪ Primary sore – starts as ▪ Monocytosis and
manifestations lesions a small papule → red neutropenia with
▪ Chronic itchy vesicle → granulocytopenia
ulceration of the ulcerate within 1-4 ▪ Infection of the
exposed skin weeks then disappear bone marrow
areas or leave and open ▪ Headache
weeping sore ▪ Undulant fever
▪ Secondar lesion – ▪ Hepatosplenomeg
invasion of mucous aly
membrane; before ▪ Black fever

and after the initial
sore has healed; they
are usually painful
and may cause
deformity of the nose
and may destroy
cartilages of the
septum
Laboratory ▪ Skin scrapings ▪ Puncture of the ▪ Blood smear
diagnosis (Wright’s or ulcerated part ▪ Lymph node
Giemsa stain) ▪ Demonstration of the aspirate
▪ Impression parasite from smears
▪ Bone marrow
smears or tissue prepared from lesion puncture
sections scrapings (particularly
prepared from in the edge)
active lesions;
biopsies should
be obtained
from elevated,
inflamed
margins of the
lesion
Diagnostic Demonstration of amastigote in macrophage in Giemsa stained smears
stage from aspirates or biopsies from lesion
Treatment and ▪ Treat adequately all human cases harboring the parasite with
Prevention antimony
▪ Secondary lesions are treated by topical disinfection, curettage or
excision of redundant tissues (L. braziliensis)
▪ Eradicate breeding ground of sandflies
Other ▪ Novy-McNeal-Nicolle medium
Laboratory ▪ Montenegro test – in vivo skin test
methods
Genus Trypanosoma:
Exists in two forms:
✓ Epimastigote
✓ Trypomastigote
A. Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense
▪ Causative agent of African sleeping sickness
▪ Transmitted by the bite of infected tse tse flies of the Genus Glossina
▪ Characterized by an acute phase in which peripheral blood and lymph
nodes are invaded, followed by a chronic phase in which the central
nervous system is invaded, resulting in meningoencephalitis
▪ Coma state develops, commonly known as “sleeping sickness”

LIFE CYCLE: (Trypanosoma brucei)
1. During a blood meal on the mammalian host, an infected tsetse fly (genus
Glossina) injects metacyclic trypomastigotes into skin tissue.
2. The parasites enter the lymphatic system and pass into the bloodstream
3. Inside the host, they transform into bloodstream trypomastigotes, are carried to
other sites throughout the body, reach other body fluids (e.g., lymph, spinal fluid),
and continue the replication by binary fission
4. The entire life cycle of African trypanosomes is represented by extracellular
stages.
5. The tsetse fly becomes infected with bloodstream trypomastigotes when taking
a blood meal on an infected mammalian host
6. In the fly’s midgut, the parasites transform into procyclic trypomastigotes, multiply
by binary fission, leave the midgut, and transform into epimastigotes
7. The epimastigotes reach the fly’s salivary glands and continue multiplication by
binary fission
8. The cycle in the fly takes approximately 3 weeks.
9. Rarely, T. b. gambiense may be acquired congenitally if the mother is infected
during pregnancy.

B. Trypanosoma cruzi
▪ Causative agent of American trypanosomiasis or Chaga’s disease
▪ Most common vectors are triatomid bugs, kissing bugs, or reduviid bugs
o Demonstration of trypomastigote in thin and thick smears prepared
from peripheral blood or CSF (C-shaped trypomastigotes)
o Xenodiagnosis:
✓ Use trypanosome free laboratory bred bug to bite infected
individual
✓ Examine for the intestinal content of bug for metacyclic
trypanosomes
LIFE CYCLE: (Trypanosoma cruzi)
1. An infected triatomine insect vector (or “kissing” bug) takes a blood meal
and releases trypomastigotes in its feces near the site of the bite wound.
2. Trypomastigotes enter the host through the bite wound or intact mucosal
membranes, such as the conjunctiva
3. Inside the host, the trypomastigotes invade cells near the site of
inoculation, where they differentiate into intracellular amastigotes
4. The amastigotes multiply by binary fission and differentiate into
trypomastigotes, and then are released into the circulation

5. Trypomastigotes infect cells from a variety of tissues and transform into
intracellular amastigotes in new infection sites.
6. Clinical manifestations can result from this infective cycle.
7. The bloodstream trypomastigotes do not replicate (different from the
African trypanosomes).
8. Replication resumes only when the parasites enter another cell or are
ingested by another vector.
9. The “kissing” bug becomes infected by feeding on human or animal
blood that contains circulating parasites
10. The ingested trypomastigotes transform into epimastigotes in the vector’s
midgut
11. The parasites multiply and differentiate in the midgut and differentiate into
infective metacyclic trypomastigotes in the hindgut
12. Other less common routes of transmission include blood transfusions,
organ transplantation, transplacental transmission, and foodborne
transmission (via food/drink contaminated with the vector and/or its
feces).
Trypanosoma species
Differences T. brucei gambiense T. brucei rhodesiense T. cruzi (Chaga, 1909)
(Dutton, 1902) (Stephen and
Fantham, 1910)
Diseases ▪ Mild African ▪ East African ▪ South American
sleeping sickness sleeping sickness trypanosomiasis
▪ West African ▪ Rhodesian ▪ Chaga’s disease
sleeping sickness trypanosomiasis
▪ Gambian
Trypanosomiasis
Distribution Central and West East and South Africa ▪ Asia minor
Africa ▪ Middle and
Southwest Asia
▪ North Africa
▪ Europe and
Central America
Vector Glossina palpalis (tse Glossina palpalis ▪ Triatomid bug
tse fly) Glossina morsitans (Panstrongylus
Glossina tachinoides megistus)
▪ Kissing bug
▪ Assassin bug
Mode of Insect bite after the complete cycle in the ▪ Bug bite and
Transmission insect defecates on the
wound
▪ Blood transfusion
▪ Sexual and
congenital
transmission

Infective stage Promastigotes
Main Habitat Blood, skin, lymph glands, cerebrospinal Reticuloendothelial
fluid, brain cells, central nervous
system
Pathogenesis ▪ Parasitemia with different waves ▪ Romana’s sign
▪ Winterbottom’s sign (edema of the
▪ Kerandel’s sign eyelids)
▪ Signs and Symptoms: ▪ Chagoma (initial
✓ Chancre lesion)
✓ Cervical lymphadenopathy
✓ Recurrent high fever
✓ Headache, rash
✓ Patient develops hepatic and
renal damage, pericardial
inflammation, muscle tremors,
convulsion, and coma
▪ Gambian trypanosomiasis – CNS
symptoms; takes years to develop
▪ Rhodesian trypanosomiasis – CNS
manifestations; develop in 3 – 6 months
Diagnosis Demonstration of trypomastigote from ▪ Demonstration of

Giemsa stained thick and thin blood smears trypomastigote
as well as aspirate from chancre at the site from Giemsa
of insect bite, enlarged lymph node and stained thick and
spinal fluid thin blood smears
as well as aspirate
from chancre at
the site of insect
bite, enlarged
lymph node and
spinal fluid
▪ Demonstration of
amastigote in
macrophage
Treatment Pentamidine and Suramine soprol No drug available

Prevention 1. Proper treatment of diagnosed 1. Vector control
and Control individuals 2. Screening and
2. Eradication of breeding sites of tse tse sterilization of
flies blood transfusions

SYNTHESIS
Members of the clinically significant group of parasites located in blood and tissue
that move by means of flagella, known as the hemoflagellates, belong to the genera
Leishmania and Trypanosoma.
There are four morphologic forms of clinical significance associated with these
hemoflagellates: amastigote, promastigote, epimastigote, and trypomastigote.
Although the specific life cycle may vary, all the organisms in these two genera involve
some combination of the four morphologic forms. The transmission of all hemoflagellates
is via the bite of an arthropod vector. The major difference between the two genera is
the primary diagnostic form found in each; for Leishmania it is the amastigote and for
Trypanosoma it is the trypomastigote, with the exception of Trypanosoma cruzi, in which
amastigotes may also be found.
ASSESSMENT
QUIZ
form

REFERENCES
Inc.

LABORATORY
NAME: RATING:
Experiment No. 14
PHYLUM SARCOMASTIGOPHORA - SUBPHYLUM MASTIGOPHORA
I. Background
The organ of locomotion for class mastigophora is the flagella. Almost every species of
vertebrate can serve as host to intestinal and atrial flagellates, and a single host may harbor
several species. Special organs, such as sucking disc, axostyle, and undulating membrane, have
been developed to withstand the peristaltic action of the intestine.
Transmission from host to host is through formation of resistant cyst in many species, but in
others infection is apparently transmitted through the less hardy trophozoites. Owing to
morphologic variation, it is difficult to determine whether some species infecting humans and
lower animals are identical.
Humanity is the host of seven species, including five intestinal and two atrial parasites. The
five nonpathogenic cosmopolitan species are (1) Chilomastix mesnili, (2) Enteromonas hominis,
(3) Retortamonas intestinalis, (4) Trichomonas hominis of the intestine and (5) Trichomonas tenax
of the mouth. The pathogenic species are (6) Giardia intestinalis and (7) Trichomonas vaginalis.

1. To be able to know the morphology of mastigophora

2. To describe the life cycle of mastigophora and know the mode of transmission
3. To be familiar with the diagnostic and infective stages of these amoebae especially
subphylum mastigophora
4. Identify the subphylum mastigophora using the distinguishing marks of their diagnostic
stages

Microscope
Prepared slides
Colored plates
Reference Book

a. Draw and label:

a. Print and Label :Microscopic (HPO)
1. Giardia intestinalis cyst and trophozoite
2. Chilomastix mesnili cyst and trophozoite
3. Trichomonas vaginalis trophozoite
b. Label: Diagrammatic (Cyst and trophozoite)
1. Giardia lamblia cyst and trophozoite
2. Trichomonas hominis trophozoite
3. Trichomonas tenax trophozoite
Giardia intestinalis cyst Giardia intestinalis trophozoite
Chilomastix mesnili cyst Chilomastix mesnili trophozoite

Trichomonas vaginalis trophozoite
LABEL (Diagrammatic)
Giardia lamblia trophozoite and cyst
1. ________________________
2. ________________________
3. ________________________ 1. ________________________
4. ________________________ 2. ________________________
5. ________________________ 3. ________________________
6. ________________________ 4. ________________________
7. ________________________
8. ________________________
9. ________________________

Trichomonas hominis trophozoite
1. _______________________________
2. _______________________________
3. _______________________________
4. _______________________________
5. _______________________________
6. _______________________________
7. _______________________________
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=im
ages&cd=&ved=2ahUKEwjA6PH48pjcAhVBTJQKHY4cCPwQjxx6BA
gBEAI&url=http%3A%2F%2Fwww.biologydiscussion.com%2Fanim
als-2%2Fphylum-protozoa%2Fspecies-of-trichomonas-phylum-
protozoa%2F34326&psig=AOvVaw2SxnLSAE_gvD-
DTYT8WB7z&ust=1531462033254859)
Trichomonas tenax trophozoite
1. ______________________________
2. ______________________________
3. ______________________________
4. ______________________________
5. ______________________________
6. ______________________________
7. ______________________________
(https://www.google.com/url?sa=i&rct=j&q=&esrc=s&source=im
ages&cd=&ved=&url=http%3A%2F%2Fwww.biologydiscussion.co
m%2Fanimals-2%2Fphylum-protozoa%2Fspecies-of-trichomonas-
phylumprotozoa%2F34326&psig=AOvVaw04GQOZr_3VuGFmI_re
XwHW&ust=1531462423873350)

PRINT and LABEL (Diagrammatic)
Leishmania species (developmental forms)
Trypanosoma species (developmental forms)

1. Give the general morphology of Mastigophora

2. Describe the life cycle , MOT, diagnostic and infective stage of Mastigophora
3. In a table form differentiate the members of class mastigophora based on the characteristics of
the cyst and trophozoite
4. List and describe the laboratory methods for the isolation and identification of the different
members of mastigophora.

SECTION 5D: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
CILIOPHORA
1. Identified each member of the Protozoans of the Phylum Ciliophora, Class

Kinetofragminophorea according to morphology, and recognize stage of the
parasite being studied
confidentiality
LESSON PROPER
Members of this group of Protozoa are classified in the Phylum Ciliophora, Class
Kinetofragminophorea, which includes one-celled organisms that are provided with
short thread-like extensions of their ectoplasmic membrane (cilia) during some stages of
their life. In most of the ciliates, cilia are present during all stages of development; in
some, however, cilia are lacking in the definitive stage.
Genus Balantidium
➢ Members of this genus are exclusively parasitic in the digestive tract of

invertebrate host. As in other ciliates, asexual multiplication is accomplished by
transverse binary fission
Balantidium coli
▪ Pathogenic
▪ Habitat: lumen, mucosa, submucosa of the large intestine
▪ Mode of Transmission: ingestion of infective cysts (probably from feces of swine)
▪ Morphology:
o Trophozoite:

✓ Ovoid (sometime assumes sac shaped); usually 40 – 50 um but can
reach up to 200 um
✓ 2 kinds of nuclei: kidney-shaped macronucleus and dot-like
micronucleus
✓ Covered with cilia – enclosed in a delicate protective pellicle
covered with spiral cilia
✓ Funnel-shaped cytostome
✓ Cytoplasm: contractile vacuoles, macronucleus and micronucleus,
and food vacuoles
✓ Motility: rapid rotary motion
✓ Reproduction: transverse binary fission
o Cyst:
✓ Infective stage
✓ Mononucleated, spherical and oval in shape
✓ Thick refractile cell wall (refractile double wall enclosing cilia)
▪ Life cycle:
o Similar with E. histolytica life cycle EXCEPT:
✓ No multiplication in the cystic stage
✓ When ingested by new host, the cyst wall will be dissolved
▪ Diseases: Balantidiasis, Balantidiosis, Balantidial dysentery
▪ Pathogenicity: same with E. histolytica but in fatal cases, ulceration and
gangrene are observed
o Symptoms:
➢ Abdominal discomfort plus dysentery
➢ Organism invades tissues: submucosal lesions and hemorrhage
▪ Diagnosis:
o Direct Fecal Smear
o Concentration techniques
o Culture method (Barnet and Yarbrough’s medium, Balamuth’s medium)
▪ Prevention and Control: same with E. histolytica
▪ Treatment: Metronidazole

SYNTHESIS
Balantidium coli is the only pathogenic ciliate and is the largest protozoan
parasitizing humans. The most important key structure of identification of this protozoa is
the presence of cilia. Some individuals with B. coli infections are totally asymptomatic,
whereas others have symptoms of severe dysentery similar in amoebiasis caused by E.
histolytica.
ASSESSMENT
QUIZ
form

REFERENCES

LABORATORY
NAME: RATING:
Experiment No. 15
PHYLUM CILIOPHORA - ORDER TRICHOSTOMATIDA
I. Background
Phylum ciliophora undergoes transverse binary fission; Cyst formation also occurs, but unlike
in the phylum Sarcomastigophora, cyst formation in ciliates is purely protective, and not
reproductive (no division of nucleus or cytoplasm). Conjugation has also been documented in
Balantidium coli, the only ciliate parasite of man. However, conjugation is not a common occurrence,
and is probably not essential to the perpetuation of the species.
Characteristics of order Trichostomatida include apical or near apical densely ciliated

vestibulum; free living or parasitic in the gut of vertebrates and invertebrates.
1. To be able to know the morphology of Balanditium coli

2. To describe the life cycle of Balantidium coli
3. To know the mode of transmission Balantidium coli
4. To be familiar with the diagnostic and infective stage of Balantidium coli
5. Identify the Balantidium coli using the distinguishing mark of its diagnostic stage
Microscope
Prepared slides
Colored plates
Reference Book
a. Print and Label: Microscopic (HPO)

1. cysts and trophozoite of Balantidium coli
b. Label : Diagrammatic
1. cysts and trophozoite of Balantidium coli
cysts of Balantidium coli trophozoite of Balantidium coli
(http://aparasiteworld.blogspot.com/2010/01/balantidium-coli.html)

Trophozoite Cyst
1. ____________________________ 1. ______________________
2. ____________________________ 2. ______________________
3. ____________________________ 3. ______________________
4. ____________________________ 4. ______________________
5. ____________________________ 5. ______________________
6. ____________________________
7. ____________________________
Questions for enrichment:
1. Describe the morphology of B. coli

2. Describe the life cycle , MOT, diagnostic and infective stage and distinguishing mark/s of B. coli
3. List and describe the different laboratory methods for isolation and identification of B. coli.

SECTION 5E: MORPHOLOGY, PATHOPHYSIOLOGY, LIFE CYCLE, SPECIMENS USED FOR
APICOMPLEXA CLASS SPOROZOA
1. Identified each member of the Protozoans of the Phylum Apicomplexa, Class

Sporozoa according to morphology, and recognize stage of the parasite being
studied
confidentiality
LESSON PROPER
The phylum Apicomplexa includes parasitic protozoa that live in the body fluids
or tissues of the host. It takes its name from the apical complex that is generally present
at some stage. Most of the Apicomplexa parasitizing humans belong to the class
Sporozoa, in which the life cycle is characterized by an alternation of generations, one
sexual, and one asexual, occurring in the same host, or requiring an alternation of hosts.
In the asexual reproductive life cycle of development, multiplication is by schizogony,
while in the sexual cycle, it is by sporogony, which involves fertilization or syngamy.
Locomotion of the mature organisms is by body flexion, gliding, or undulation of

longitudinal ridges; flagella are present only on the microgametes, of some groups;
pseudopodia if present, are used for feeding, not for locomotion.
▪ Obligate intracellular parasites

▪ Life Cycle includes:
o Sexual reproduction (Sporogony): in arthropod vector
➢ The sexual cycle occurs within the intestinal lumen of the
invertebrate host

➢ END PRODUCT: SPOROZOITE (infective stage to man)
o Asexual reproduction (Schizogony): in man
➢ Asexual cycle occurs in the epithelial cells of the intestinal mucosa
➢ END PRODUCT: SCHIZONTS (infective to mosquito)
STAGES OF DEVELOPMENT:
▪ Trophozoite
➢ Feeding or growing stage in the asexual cycle
➢ Lives within the tissue cell
▪ Schizont
➢ Sporozoan body during schizogony which includes the period of initial
growth (the early schizont or presegmenter) to complete splitting up of
the nucleus with merozoite production
▪ Merozoites: late segmenters
➢ End product of schizogony in human reticuloendothelial cells
➢ Motile and escapes from the infected cells
➢ Some will infect other tissue cells going back to the trophozoite stage; the
rest will be differentiated into male and female forms (gametocytes)
▪ Gametocytes: immature sexual form
o Macrogametocyte
➢ Female gametocytes that will produce a macrogamete
➢ Mature only to be fit for fertilization
o Microgametocyte
➢ Male gametocyte that will produce a group of microgamete
▪ Gametes: mature sexual forms
o Macrogametes – female sex cells in sporozoa
o Microgametes – male sex cells in sporozoa
▪ Zygote – union of the macrogamete and microgamete; fertilized ovum
▪ Oocyst – encysted zygote
▪ Sporoblast - one of a number of bodies in many sporozoa into which the zygotes
divide and from which sporozoites are formed

▪ Sporocyst - the separated membrane that surrounds a sporoblast and
subsequently the group of sporozoites formed from this sphoroblast
▪ Sporozoite - end product of sexual multiplication of malarial parasites in
mosquito
GLOSSARY:
▪ Bradyzoites – slowly multiplying trophozoite contained in the cyst of T. gondii

▪ Exflagellation – extrusion of rapidly waving flagellum-like mircogametes from
microgametocytes
▪ Gamete – mature sex cell of plasmodia
▪ Gametocyte – immature sexual form of plasmodia (male microgametocyteor
female microgametocyte) that is present in peripheral blood
▪ Gametogony – development phase in the life cycle of malaria and coccidian
parasites in humans in which male and female gametes are formed
▪ Hypnozoite – exoerythrocytic schizont of P. vivax and P. ovale in the human liver,
characterized by delayed primary development; responsible for true relapse in
malaria
▪ Merogony – also known as schizogony; leading to the production of merozoites
in some intestinal coccidians
▪ Merozoite – product of schizogonic cycle in malaria; produced in the liver (pre-
erythrocytic cycle) and in the red blood cells (erythrocytic cycle); motile and
infects the red blood cells
▪ Oocyst – encysted form of the ookinete that occurs in the stomach wall of
Anopheles spp.
▪ Ookinete – motile zygote of Plasmodium spp; formed by microgamete
fertilization of macrogamete
▪ Paroxysm – fever, chills, sweats syndrome in malaria; spiking fever corresponds to
the release of merozoites and toxic material from the rupturing parasitized red
blood cells, and shaking chills occur during subsequent schizont development
▪ Recrudescence – increased severity of a disease after a remission or following
treatment as a result of an inadequate immune response by the host or
inadequate response to treatment

▪ Relapse - a recurrence of illness/signs and symptoms of a disease after a period
of improvement; In malaria, it is caused by the reactivation of hypnozoites in the
liver that begins a new cycle in red blood cells; occurs only in Plasmodium vivax
and P. ovale infections
▪ Schizogony – (Asexual cycle) occurs on the epithelial cells of the intestinal
mucosa producing schizonts
▪ Schizont – developed stage of asexual division of the sporozoa; ruptures to
produce merozoites
▪ Sporogony – (Sexual cycle) occurs within the intestinal lumen of the invertebrate
host. End product → sporozoite
▪ Sporozoite – slender, spindle-shaped organism; infective stage of malaria
parasites; inoculated into humans by the bite of an infected female mosquito; It
is the result of the sexual cycle in the Anopheles mosquito
▪ Tachyzoites – rapidly multiplying stage in the development of the tissue phase of
certain organisms such as Toxoplasma gondii
▪ Trophozoite – feeding or growing stage in the asexual cycle
▪ Zygote – union of the macrogamete and microgamete; fertilized ovum/ova
before cell division
A. Plasmodium species
Phylum: Apicomplexa → Class: Sporozoa → Blood species: Plasmodium
▪ Pathogenic to man
▪ Causative agent of malaria
▪ Principal vector: Anopheles minismus var. flavirostris
▪ Characteristics:
➢ Obligate intracellular parasites of blood and tissues
➢ Alternation of generations (sexual and asexual development)
➢ Alternation of host:
o Sexual cycle – female mosquito (Anopheles minimus
flavirostris)
o Asexual cycle – man

✓ sporozoites liberated into the bloodstream via bite of an infected
female mosquito;
✓ through blood transfusion
✓ vertical transmission
▪ Symptoms and Pathology:
➢ Anemia (due to massive red cell destruction), splenomegaly, joint pain
➢ Recurrent/Intermittent chills and fever (synchronized rupture of red
blood cells)
o Every 36 hours: Malignant Tertian Malaria (P. falciparum)
o Every 48 hours: Ovale Malaria (P. ovale)
o Every 48 hours: Benign Tertian Malaria (P. vivax)
o Every 72 hours: Quartan Malaria (P. malariae)
➢ Quotidian fever – is caused by the asynchronous release of merozoites
in the circulation
▪ Species:
a. Plasmodium malariae
➢ Single large compact ring or band forms
➢ Invades old RBCs
➢ Schizont with merozoites arranges around central pigment
(resembles fruit pie)
➢ Ovoid gametocytes
b. Plasmodium falciparum
➢ Small ring forms (1/6 diameter red cell), applique forms, double
nuclear dots
➢ Organisms invades all ages of red blood cells (most severe)
➢ Crescent/banana-shaped gametocytes
c. Plasmodium ovale
➢ Single compact ring
➢ Large pale red cells with Schuffner’s dots which may be oval
and fimbriated

d. Plasmodium vivax
➢ Single large ring succeeded by amoeboid form in pale large
red cell
➢ Schuffner’s dot (condensed hemoglobin) in red cells
➢ Only reticulocytes are invaded
➢ Round gametocyte
▪ Note:
✓ Plasmodium vivax infection is most widely distributed and most
prevalent worldwide
✓ Plasmodium falciparum infection is most likely fatal
o Cerebral malaria: red cells, organisms and pigment can block
the brain vessels
o Blackwater fever: sudden massive intravascular hemolysis
resulting to hemoglobinuria
1. Microscopic identification of the malarial parasites in thick and thin
blood smears stained with Giemsa or Wright’s stain is still important in
making the definitive diagnosis and remains the gold standard
method.
2. Collection of specimen must be prior to fever spike
3. Bone marrow (through sternal puncture)
4. Malaria RDTs (Rapid Diagnostic Tests):
o Plasmodium LDH – produced by both sexual and asexual stages
and can distinguish between P. falciparum and non-P.
falciparum species
Examples:
1. Diamed Optimal IT
o Immunochromatography – detects Plasmodium-specific

antigens; these target antigens are called HRP II (Histidine-rich
protein)
Examples:

1. Paracheck Pf test
2. ParaHIT f test
5. Serological tests (to detect the presence of malarial antibodies)
▪ Life Cycle:
I. Asexual phase in man (SCHIZOGONY/MEROGONY)

A. Pre-erythrocytic/Exo-erythrocytic schizogony:
1. Begins with the inoculation of the infective sporozoites to man during a
mosquito blood meal

2. Within ½ hour, they are carried through blood circulation into the liver
parenchymal cells where they undergo nuclear and cytoplasmic
division and develop into pre/exo-erythrocytic schizonts
3. Schizonts rupture producing exoerythrocytic merozoites that reinvade
liver cells, while other invade the RBCs
4. In the RBC, merozoite develops into trophozoite
5. In P. vivax and P. ovale, sporozoites develop into hypnozoites which
remain dormant for years in the hepatocytes. At a predetermined
time, the hypnozoites begin to grow and undergo exo-erythrocytic
schizogony releasing merozoites that invade RBCs causing a
recurrence of the malaria attack
B. Erythrocytic schizogony:
1. The trophozoite further matures into schizont, then divide into
erythrocytic merozoites
2. RBC ruptures releasing merozoites into the bloodstream
C. Gametogony:
1. After 2 – 3 eryhtrocytic generations, Gametocytogenesis begins
2. Some of the merozoites do not form schizont, but rather micro- and
macrogametocytes which are infective to the mosquito
II. Sexual phase in mosquito (SPOROGONY)
1. The male and female gametocytes sucked in by the mosquito undergo
maturation and differentiate into micro- and macrogametes
2. The microgamete exflagellates and fertilizes the macrogamete producing
a zygote as a result of fertilization
3. Ookinete penetrates the stomach wall and forms an oocyst
4. Within the oocyst, numerous sporozoites are formed
5. Oocysts grows and ruptures releasing sporozoites
6. Sporozoites migrate through tissues to the salivary glands
Plasmodium species
Plasmodium Plasmodium Plasmodium Plasmodium
falciparum malariae ovale vivax
Disease Malignant Quartan Ovale malaria Tertian malaria
malaria malaria

Paroxysm 36- 48 hours 72 hours 48 hours 48 hours
cycle
Infected RBCs Not enlarged Not enlarged Sometimes Enlarged
enlarged;
frequently oval
with ragged
margins
Appearance of Normal; Normal Enlarged; Enrlaged;
RBC size multiply approximately maximum size
infected red 20% or more of may be 1 – 2
blood cells are infected RBCs times normal
common are oval RBC diameter
and/or
fimbriated
(border has
irregular
projections)
Number of 6 – 32 6 – 12 6 – 14; average 12 – 24;
merozoites (average is (average is 8); is 8 average is 16
20 – 24) “rosette”
schizonts
Schuffner’s - - + +
stippling (Maurer’s dots (Ziemann’s (James’ dots; (Schuffner’s
(precipitated occasionally dots rarely present in all dots; present
Hb) seen) seen) stages except with all stages
early ring except in early
forms) ring forms)
Parasite Young rings are Rounded, Rounded, Irregular,
cytoplasm small, delicate, compact compact ameboid
often with trophozoites trophozoites; trophozoites;
double with dense occasionally has “spread
chromatin cytoplasm; slightly out”
dots; band-form amoeboid; appearance
gametocytes trophozoites growing
are crescent- trophozoites
shaped or have large
elongated chromatin
mass
Trophozoite Accole or Band Red cell Amoeboid
Applique forms containing
May have trophozoite
multiple rings may have
fimbriated
edges
Appearance of Black; coarse Dark brown, Dark brown, Golden brown,
parasite and coarse, conspicuous inconspicuous
pigment conspicuous in conspicuous
gametocytes
Shape of Sausage or Round Round Round
gametocyte crescent-
shaped

Stages seen in Rings and/or All stages; wide All stages All stages; wide
circulating gametocytes; variety of range of
peripheral other stages stages usually stages may be
blood develop in not seen; seen on any
blood vessels relatively few given film
of internal rings or
organs but are gametocytes
not seen in generally
peripheral present
blood EXCEPT
in severe
infection
Multiple YES NO NO RARE
infections
Others Highy mortality Rarely fatal Least common Most common
Rarely fatal Rarely fatal
May cause May cause
relapses relapses
B. Babesia species
Phylum: Apicomplexa → Class: Sporozoa → Blood species: Babesia
▪ Pathogenic: Babesia microti
▪ Definitive host: Animals (Deer)
▪ Transmission: man infected by bite of a tick that belong to genus Ixodes
(intermediate host); can be transmitted through blood transfusion
▪ Infective stage: trophozoites liberated via the bite of deer tick
▪ Morphology:
o An obligate intracellular parasite (seen inside of an RBC measuring
about 2 – 4 um)
o Pear-shaped
o Usually in pair or tetrads (resembling “maltese cross” appearance)
▪ Symptoms and Pathology: symptoms resemble Malaria
o Headache and fever
o Hemolytic anemia with hemoglobinuria in immunocompetent
host
▪ Diagnostic stage: demonstration of characteristic ring forms in Giemsa-
stained blood smears (thick and thin smear)

C. Coccidians
➢ Coccidian parasite are members of the Class Sporozoa in the Phylum
Apicomplexa. The subclass Coccidia includes species of Toxoplasma,
Isospora, Sarcocystis, Cryptosporidium, and Cyclospora
➢ Life cycle:
o Schizogony (Asexual) in variety of nucleated cells
o Sporogony (Sexual) in intestinal mucosa of definitive host: infective
oocyst are excreted in the feces
➢ Classification:
✓ Intestinal Coccidian:
− Prevalent in AIDS patient/immunocompromised persons
− Infective stage: oocysts
− Diagnostic stage: oocysts demonstrated in feces
✓ Tissue Coccidian
I. Intestinal Coccidian
a. Cryptosporidium parvum
▪ Important opportunistic infection in AIDS patients
➢ Ingestion of oocysts from food or water contaminated with
animal feces
➢ Oral-anal route
➢ Direct contact with infected individual or animal
▪ Method of infection:
➢ Upon ingestion, sporozoites released from oocyst
➢ Develop in brush border of intestinal epithelial cells
➢ Sporulated oocysts, containing 4 sporozoites each (no
sporocysts), are passed in feces
➢ Infective oocysts are transmitted via fecal-oral route
▪ Disease: Cryptosporidiosis
➢ Causes intestinal infection: associated with watery, frothy
diarrhea with oocysts shed in feces
➢ Causes chronic diarrhea in immunocompromised person

➢ Acute self-limiting diarrhea
▪ Diagnosis:
➢ Sugar floatation technique
➢ Modified acid fast stain in feces:
o Red spherical bodies, four sphorozoites
o Oocysts are stained red against blue background
o Average size: 4 – 6 um
b. Cyclospora cayetanensis
− infective oocysts ingested in contaminated food and water
− outbreaks have been associated with contaminated berries
▪ Clinical disesase: indistinguishable from cryptosporidiosis
▪ Diagnosis:
o Modified AFS
➢ Oocysts stain from light pink to deep red (acid-fast
variable)
➢ Average size: 8 – 10 um (larger than C. Parvum)
c. Isospora belli
▪ Mode of Transmission: ingestion of sporulated oocysts in fecally
contaminated food or water
▪ Definitive host: Humans
▪ Habitat: small intestines of man
▪ Morphology:
o Immature oocyst
➢ 20 – 33 um by 10 – 19 um
➢ Elongately ovoidal in shape with one end narrower than
the other
o Mature oocyst
➢ 29 um by 14 um
➢ Contains 2 sporocyst, each containing 4 sporozoites
▪ Disease: Human Coccidiosis
➢ Often asymptomatic and self-limiting

➢ Symptoms range from mild gastrointestinal distress to severe
dysentery
o In mild cases: mild abdominal pain and mucoid diarrhea
o In severe cases: severe abdominal cramps with milky,
watery diarrhea
▪ Diagnosis
o Demonstration of oocysts in feces (transparent containing 1-2
sporoblast)
o Modified AFS
➢ Sporoblasts and/or sporocysts stain deep red
➢ Oocysts are ellipsoid with blunt ends
➢ Average size: 30 by 12 um
II. Tissue Coccidian
Toxoplasma gondii
▪ Prevalent in AIDS patients/immunocompromised persons
▪ Definitive host: cat
▪ Intermediate host: humans
▪ Habitat: intracellular obligate parasite of endothelial cells,
mononuclear leukocytes, body fluids, and tissue of the host
▪ Transmission:
➢ Accidental ingestion/inhalation of oocysts from cat feces
➢ Ingestion of undercooked meat or oocysts from cat feces
➢ Transplacental
➢ Organ transplants
▪ Morphology:
➢ Crescent appearance in tissue fluids
➢ Tissue stage in man: both cause infection
o Tachyzoites: fast rapid, multiplication, acute phase
o Bradyzoites: slow proliferation, chronic phase
▪ Pathogenesis
➢ Toxoplasma divide in tissues of man as tachyzoites
➢ Pseudocysts (group of bradyzoites) are also formed

➢ Female who first to acquire the infection during pregnancy may
transmit the infection to embryo resulting in fetal death, or
mental retardation on newborn, or blindness in later life
▪ Disease: Toxoplasmosis
o Major cause of encephalitis in AIDS patients
o Acquired toxoplasmosis:
➢ Appears after the infection and regional lymph node
invasion
➢ Parasite is blood borne to many organs where
intracellular multiplication takes place
o Major cause of congenital toxoplasmosis among the newborns:
➢ Congenital infection causes birth defects and mental
retardation
▪ Diagnosis
o Serological diagnosis: EIA and IFA – for detecting neonatal
toxoplasmosis
o Sabin-Feldman Dye test:
➢ Methylene blue staining of tachyzoites inhibited by prior
addition of patient serum containing antibodies of
Toxoplasma
D. Microsporidia
➢ Newest group of obligate intracellular parasite
Enterocytozoon bieneusi (Encephalitozoon intestinalis)
▪ The most common microsporidia causing enteritis among patients with

AIDS
▪ The organism is very small measuring about 1.5 – 4 um
▪ Characteristic feature: spores containing a polar tubule, used to inject
infective spore content into the host cells
➢ Not certain; most likely by ingestion of spores

➢ Inhalation of spores, ocular exposure, and sexual intercourse may
also be route of transmission
▪ Clinical disease
➢ Similar with Cryptosporidiosis
➢ Spores are very resistant
▪ Diagnosis:
o Electron Microscopy – necessary to speciate
o Serological testing
o Modified Trichrome stain:
➢ Concentration must be 10x higher that traditional trichrome
stain
➢ Performed on unconcentrated specimen
➢ Spore walls stains bright pink; background stains green or
blue (depending on the couterstain)
SYNTHESIS
Most of the Apicomplexa parasitzing man has a life cycle that is characterized by
an alteration of generations, one sexual, and one asexual, occurring in the same host, or
requiring an alternation of hosts. As with all parasites, the proper identification of malaria
and babesiosis is crucial to ensure that the patient is adequately treated when necessary.
Plasmodium and Babesia spp. have morphologic forms that may look similar. However,
because not all species typically show all the morphologic forms in the peripheral blood,
coupled with the fact that other morphologic forms look different (e.g., mature schizonts,
gametocytes) and whether pigment is produced, allow accurate speciation of the
malarial organism and differentiation of malaria from babesiosis.
The miscellaneous protozoa (Coccidians) have morphologic similarities (e.g., the

oocysts of Isospora and Sarcocystis) and distinct differences. When screening suspected
samples, attention to organism size, shape, andstructural details is imperative to identify

parasites correctly. Note that most of these coccidians are opportunistic especially
among immucompetent individuals. Among all of the coccidian parasites, T. gondii is the
most clinically significant implicated in causing neonatal toxoplasmosis.

ASSESSMENT
RESEARCH ACTIVITY:
Instruction: Give a brief discussion on the following questions. Indicate the reference
used. (20 POINTS)
1. Explain how does hemoglobin SS (sickle cell anemia) and G6PD deficiency
confers resistance against malaria?
QUIZ
form

REFERENCES
Inc.

LABORATORY
NAME: RATING:
Experiment No. 16
PHYLUM APICOMPLEXA - CLASS SPOROZOEA
I. Background
The phylum name comes from the apical complex that is present at some stage of the parasite
and is made up of elements such as the polar ring, rhoptries, micronemes, conoid and
subpellicular microtubules, which are visible under the electron microscope.
In the class Sporozoea, the life cycle is characterized by an alternation of generations – one
sexual and one asexual - occurring in the same host or in another host. In asexual reproduction,
multiplication is by schizogony and in the sexual cycle, multiplication is by sporogony.
Suborder Eimeriina: Isospora, Sarcocystis, Toxoplasma, Cryptbsporidium, Cyclospora
Suborder Haemosporina: Plasmodium spp
Subclass Piroplasmia: Babesia

1. To be able to know the morphology of class sporozoa

2. To describe the life cycle of class sporozoa
3. To know the mode of transmission of class sporozoa
4. To be familiar with the diagnostic and infective stages of species under class sporozoa
5. To identify the species under class sporozoa using the distinguishing marks of their
diagnostic stages and correct laboratory methods

Microscope stain Colored plates Books
Glass slide Giemsa Lancet Cotton balls

Methanol

a. Procedure:
A. Preparing thick and thin blood film on the same slide
1. The second or third finger of the left hand is generally selected for blood
collection. Wipe the finger with 70% alcohol and allow to dry.
2. Pierce the finger with a lancet at the side of the ball of the finger. Wipe off
the first drop of blood with a dry cotton ball.
3. Gently squeeze the finger until a second drop of blood form. Touch this drop
near the end of a pre-cleaned slide. Spread the drop of blood with the corner
of another slide to make a circle about 1 cm in diameter. This is the thick
blood film.
4. Put a new drop of blood on the margin left of the thick film. Place the slide
on the table and approximate the end of the second clean slide to be used as
a spreader, to the drops until a continuous film is formed at the edge.
5. Holding this second slide at an angle of 45 o, push it forward with a rapid but
not too brisk a movement.
6. Using soft lead pencil, label the slide approximately on the film. Allow thick
and thin film to dry.
B. Procedure for Staining the Blood film
1. Immerse the thick film in tap water for 2 -3 minutes.
2. Fix the film with methanol for 2 - 3 minutes.
3. Dry then stain with Giemsa for 30 - 60 minutes.
4. Wash with tap water
5. Dry, then examine microscopically
Note:
This is used for the examination of malaria and filaria.
b. Draw and label:

A. Print and Label: Diagrammatic
1. Plasmodium vivax - Erythrocytic stages
2. Plasmodium ovale - Erythrocytic stages
3. Plasmodium falciparum - Erythrocytic stages
4. Plasmodium malariae - Erythrocytic stages
5. Life cycle of Plasmodium - showing schizogony and sporogony
6. Life cycle of Toxoplasma gondii
7. Life cycle of Cryptosporidium parvum

ERYTHROCYTIC STAGES OF THE DIFFERENT PLASMODIUM SPECIE

Life cycle of Plasmodium - showing schizogony and sporogony
Life cycle of Toxoplasma gondii

Life cycle of Cryptosporidium parvum
1. Give the general morphology of Class sporozoea

2. Describe the life cycle , MOT, diagnostic and infective stage of Sporozoea
3. List and describe the different laboratory methods for isolation and identification of the
different species of class Sporozoea
4. In a table form, differentiate the different Plasmodium specie

SECTION 6: ECTOPARASITES OF MEDICAL IMPORTANCE
▪ To appreciate the characteristic and role of ectoparasites in Medical

Parasitology
LESSON PROPER
Of all the major divisions of phyla which make up the animal kingdom, Phylum
Arthropoda is certainly one of the most important. These are bilaterally symmetrical
invertebrate animals with outer coverings or exoskeletons. The arthropods range in size
from the Atlas moth with a wingspread of 30.48 cm, to the small follicle mite, less than
1/250 of an inch long. Some arthropods are parasitic while most are non-parasitic.
Some prefer to live in highly organized and complex environments in which each
member contributes something to others in asymbiotic relationship
Disease Causation:
▪ They may take up residence as temporary or permanent occupants

▪ They may cause the disease themselves by growing in (infection) skin or hair of
their host (infestation)
▪ They may transmit diseases as a mechanical transfer agents (house flies,
cockroaches)
▪ They may also transmit diseases via blood meal or excrement contamination of
bite wounds (as biologic vectors)
▪ Some arthropods affect humans by injecting venom (spiders, scorpions)
Arthropods that are Biological Vectors of Important Human Pathogens
Class: Order Arthropod Taxon Disease/Parasite Relationship/Commen

Transmitted ts
CRUSTACEA: Cyclops spp. Dracunculus Intermediate Host
COPEPODA – these medinensis
are small, graceful
and symmetrical Gnathostoma 1st Intermediate Host
animals with the spp.
head and first two
Diaptomus spp. Diphyllobothriasis
thoracic segments
Cyclops spp. Sparganosis
fused into a
(D. latum)
cephalothorax, and
a slender abdomen
of 3 – 5 segments

CRUSTACEA: Freshwater crabs, P. westermani 2nd Intermediate Host
DECAPODA Crayfishes
DIPLOPODA Julus spp. H. diminuta Some produce
Fontaria virginiensis vesicating agents
(material that irritate
the skin)
ARACHNIDA: Spotted fever – R. rickettsi Through the bite of
PARASITIFORMES Q fever – C. burnetti tick
(ticks and gamasid Tularemia – F. tulerensis
mites) Babesiosis – B. microti and B. divergens
(Ixodes spp.)
INSECTA: Trichodectes canis D. caninum Man is infected by
MALLOPHAGA (dog louse) swallowing infected
(chewing lice) louse
INSECTA: ANOPLURA Pediculus Typhus fever – R. prowazeki

(sucking lice) humanus humanus Trench fever – R. Quintana
▪ Pediculus (body louse) Epidemic relapsing fever – B. recurrentis
humanus var
capitis (head
louse)
▪ Phthrius pubis
(crab louse)
Triatoma spp. Chaga’s disease Inoculated through
INSECTA: HEMIPTERA Panstrongylus spp. (T. cruzi) feces of bugs
(True bugs) Rhodnius spp.
Anopheles spp. Malaria Definitive host
Anopheles spp. B. malayi
Mansonia spp.
Aedes spp.
Culex spp. W. bancrofti
Aedes spp.
Anopheles spp.
INSECTA: DIPTERA Mansonia uniformes
Culicoides spp. M. perstants
M. streptocerca
M. ozzardi
Simulium spp. O. volvulus
(black flies, coffee flies) M. ozzardi
Phlebotomus spp. Leishmania spp.
Lutzomyia spp.
Chrysops fly (mango flies) Loa loa
Chrysops fly (deer flies) F. tularensis
Glossina spp. T. brucei spp.
Xenopsylla Y. pestis Acquired through flea
cheopis R. typhi bite

Xenopsylla H. diminuta Acquired by
INSECTA: cheopis swallowing infected
SIPHONAPTERA (and other rat flea
(fleas) fleas)
Ctenecephalides D. caninum
canis
C. felis
Pulex irritans
INSECTA: Many species H. diminuta Acquired by
COLEOPTERA swallowing infected
(beetles) flea
INSECTA Few species of H. diminuta
COLEOPTERA: grain moths
(moths and
butterflies)
SYNTHESIS
In considering arthropods as biologic vectors of human pathogens, one should

remember that, in many instances, these microorganisms are so well adapted to the
arthropod host that they produce little or no tissue damage. Transmission to man and
other higher animals is accidental and frequently incidental to the perpetuity of the
microorganism. Moreover, many other species of microorganisms are normal parasites of
the arthropod and are not known to be transmissible to man or other vertebrates.
ASSESSMENT
QUIZ
form

REFERENCES

Module Evaluation Questionnaire
The learner’s feedback is vital to us. Taking into account your assessment and impression will
help us enrich the content enhance the quality your learning engagement with us.
From this view, we would appreciate if you could spend some time completing this evaluation by
checking the column you think is appropriate and then providing a qualitative response to the
questions raised in this form.
The questionnaire is anonymous and though your participation is voluntary, your utmost
cooperation is encouraged.
Once completed the results of these questionnaires will be analysed and an overview compiled
which will be reported to the next cohort of students in the module handbook. The overview will
also be used to inform discussion at programme team conference.
INDICATORS Strongly Moderately Slightly Disagree Strongly

agree agree agree agree
The Module
a) was effectively designed
b) had clear learning outcomes
c) was well organized
d) contained relevant information
e) had clear images
f) had sufficient parts
g) had lessons that related to life
experiences
The Assessment
a) rubrics were clear
b) instructions were comprehensive
c) was sufficiently challenging
d) was aligned to the lessons
e) was done within the prescribed time
f) had enriched my knowledge about the
lessons
g) were of different types
h) contained critical thinking questions
What do you like most about the module?
What could have been improved on the module?

What other things you suggest to improve the module?
How satisfied are you with the module?
Thank you.


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Clinical Parasitology

Compiled and Prepared by

Parasitology is an essential component of clinical laboratory medicine.

In spite of the many advances in technology that has been developed in

A Self-regulated Learning Module 3

ABOUT THE MODULE

A Self-regulated Learning Module 4

A Self-regulated Learning Module 5

A Self-regulated Learning Module 6

A Self-regulated Learning Module 7

Teresa N. Villanueva, RMT, MACT

A Self-regulated Learning Module 8

2 Continue Section 1 Introduction to Online Lecture, Assessment & Quiz

3 Section 2: Nematose Intro Online Lecture: Assessment & Quiz

A Self-regulated Learning Module 9

Section 5B: Mastigophora (Flagellates) ............................................................................................ 237

Section 5C: Mastigophora (Hemoflagellates) ................................................................................... 246

Experiment no. 14: Phylum Sarcomastigophora (Subphylum Mastigophora) ........................... 256

Section 5D: Ciliophora (Ciliates) ....................................................................................................... 262

Experiment no. 15: Phylum Ciliophora....................................................................................... 265

Section 5E: Apicomplexa (Sporozoa) ................................................................................................ 268

Experiment no. 16: Phylum Apicomplexa (Class Sporozoa) ....................................................... 284

Section 6: Ectoparasites ......................................................................................................................... 289

A Self-regulated Learning Module 10

A Self-regulated Learning Module 11

COURSE LEARNING OUTCOMES:

At the end of this session, the students must have:

1. used the basic terminologies and theories related to Parasitology

1. Parasitology 11. Temporary parasite

A Self-regulated Learning Module 12

A Self-regulated Learning Module 13

A Self-regulated Learning Module 14

A Self-regulated Learning Module 15

A Self-regulated Learning Module 16

F. DISEASE PROCESSES and SYMPTOMS

A Self-regulated Learning Module 17

G. PREVENTION and CONTROL

III. LABORATORY METHODS FOR DIAGNOSIS

A. SPECIMEN FOR DIAGNOSIS

A Self-regulated Learning Module 20

Thick Blood Film:

✓ Infected erythrocytes are counted in relation to a predetermined number of

# of parasites/µl = # of parasites x 8000

# of parasites/µl =# of parasites counted x 40

• If parasites are <10, 500 WBCs should be counted:

# of parasites/µl = # of parasites counted x 16

A Self-regulated Learning Module 21

Thin Blood Film:

+ = 1–10 per 100 thick fields

++ = 11-100 per 100 thick fields

+++ = 1–10 per thick field

++++ = >10 per thick field

C. COLLECTION OF FECAL SAMPLE

A Self-regulated Learning Module 22

TYPES of FECAL SPECIMEN

1. Liquid – either diarrheic or saline-purged

A Self-regulated Learning Module 23

1. To detect the presence of intestinal parasites

D. EXAMINATION OF FECAL SAMPLE

A Self-regulated Learning Module 25