Protozoal

infection
DR. SAJINI SOUDA
 Learning Objective
 Give examples of protozoa, and the main types of infections
caused by them.
 Describe the life cycles of medically important protozoa found in
Botswana
 Describe the causative pathogens, life cycle; mode of infection;
epidemiology; treatment (especially within Botswana) of the
malaria parasite.
 Summarise the pharmacological approaches to prophylaxis and
treatment of the major protozoal infections.
 Definition
 Parasite: An organism that spends a significant portion of its life in or
on the living tissue of a host organism and causes harm to the host
without immediately killing it.

 Infection: invasion of endoparasites

 Infestation: external parasitism of ectoparasites such as arthropods.

 Definitive host: harbors the adult or sexual stage.

 Intermediate host: part or all of the larval or asexual stage takes place
 Protozoa
• Means "first animal“. They are the simplest and primitive of all animals.
• Unicellular eukaryotes: lack cell wall, contain membrane-bound
organelles, including one or more nucleus
• Size ranges from 10–50um, some up to 1 mm, seen under a microscope.
• Protozoans exhibit mainly two forms of life
• free-living (aquatic, freshwater, seawater)
• parasitic (ectoparasites or endoparasites)
• Body protoplasm is differentiated into an outer ectoplasm and an
inner endoplasm.
• Locomotor organs are fingers like pseudopodia, whip-like flagella,
hair-like cilia or none.
• Respiration occurs by diffusion through the general body surface.
Protozoa characteristics……………
Food obtained by:
• absorption through the ectoplasm–diffusion/active transport
• surround food and engulf it (phagocytosis)
• openings or “cytostomes" into which they sweep food
Digest food intracellularly in stomach-like compartments - vacuoles
Excretion occurs through the general body surface
• temporary opening in the ectoplasm or
• through a permanent pore cytopyge.
Usually exists in the trophozoite or cystic stage
Encystment - to resist unfavorable conditions of:
food, temperature, moisture.
helps in dispersal.
 5:10

s://youtu.be/Aa0cvmsD_2Q
https://youtu.be/Aa0cvmsD_2Q
 Reproduction
Reproduce asexually or sexually
 Asexual reproduction by:
❖ binary fission
❖ schizogony
Schizogony: multiple fission
o Nucleus undergoes multiple divisions
o Portions of cytoplasm then concentrate around each nucleus.
o Cell separates into multiple daughter cells.

 Sexual reproduction: Sporogony

Plasmodium

 The life cycle is often complicated with alternation of asexual and sexual phases (alternation of generation).
Microsporidium
 LO1: Give examples of the range of conditions resulting from protozoal infections, and the
main types of organism responsible
Features of medically important protozoa
Location Species Mode of transmission Disease
Entamoeba histolytica Ingestion of cysts in food Amoebiasis
Cryptosporidium spp. and water Cryptosporidiosis
Cystoisospora belli Isosporiasis
Intestinal tract Cyclospora cayetanensis Cyclosporiasis
Microsporidia Microsporidiosis
Giardia intestinalis Giardiasis
Balantidium coli Balantidiasis
Urogenital tract Trichomonas vaginalis Sexual Trichomoniasis
Babesia microti Ixodes scapularis(tick) Babesiosis
Trypanosoma spp.
Trypanosomiasis,
T. cruzi Reduviid bug
Chagas disease
T. gambiense, Tsetse fly
Sleeping sickness
T. rhodesiense
Leishmania spp.
Visceral leishmaniasis
L. donovani complex
(kala-azar)
L. tropica, L. major, Sandfly
Cutaneousleishmaniasis
Blood and tissue L. mexicana
Mucosal leishmaniasis
L. Viannia braziliensis
Plasmodium spp.
P. vivax, P. ovale, P. malariae Anopheles Mosquito Malaria
P. falciparum, P.knowlesi
Toxoplasma gondii Ingestion of cysts in raw
meat; contact with soil
Toxoplasmosis
contaminated with cat
faeces
 World annual rates of morbidity and
mortality

Infections Disease Deaths

(millions) (millions) (thousands)

Protozoa Malaria 800 150 1500

Amoeba 480 50 100
Toxoplasma 40 10
Trypanosoma 24 1.2 60
Common parasite isolated from Botswana
Number of Percent of All
Parasite Positive Positive
Specimens Microscopy
(n=297)

Isospora spp. 113 36.9%

Cryptosporidium
spp. 99 32.4%
Giardia lamblia 34 11.1%
Taenia spp. 27 8.8%
Trichomonas
hominis 20 6.5%
All other parasites 13 4.3%
JS Rowe et al 2010
 Entamoeba histolytica
• A pseudopod-forming nonflagellate amoebic protozoa
• Move by means of flowing cytoplasm with the production of
pseudopodia
• Humans are the only host
• Found in human colon

Exists in two forms-

• Cysts (formed stool) –infective form
• Trophozoites (diarrhoel stools)
• Both forms are passed in feces.
• Cysts ingested from faecally contaminated food or water.
• Sexual transmission through oral–anal sexual practices
 Clinical Manifestations of Entamoeba Infection
Asymptomatic Colonization Lab diagnosis :

• E. histolytica , E. dispar 1.Microscopy

Intestinal Amebiasis and Its Complications 2.Antigen detection

• Amebic dysentery/ acute necrotizing colitis 3.Serologic tests

• Ameboma 4.PCR

• Toxic megacolon 5.CT for abscess and granulomas

• Peritonitis – Intestinal perforations 6.Colonoscopy

• Cutaneous amebiasis Treatment

• Luminal infection: iodoquinol,
• Rectovaginal fistulas
paromomcyin, and diloxanide
Extraintestinal Amebiasis furoate. Nitazoxanide
• Amebic liver abscess (hepatitis) • Invasive amebiasis (colitis, liver
• Splenic abscess, Brain abscess , Empyema abscess) nitroimidazole -
metronidazole and tinidazole
• Pericarditis
.
 Amebic liver abscess
(Reproduced from Petri WA Jr. Recent advances in
amebiasis. Crit Rev Clin Lab Sci. 1996;33:1. Copyright Lewis
Publishers, an imprint of CRC Press.)

https://www.youtube.com/watch?v=Aa0cvmsD_2Q
Amebae that infect the human gastrointestinal tract
 Free Living Amoeba
Acanthamoeba culbertsoni
• Contaminated water
• Keratitis, Uveitis in contact lens users.
• Skin lesions – ulcers, hard nodules.
• Granulomatous dissemination- lungs
• Granulomatous necrotizing encephalitis.

Naegleria fowleri: Brain eating amoeba

• Primary Amoebic Meningoencephalitis.
• Fresh water

Balamuthia mandrillaris
• Granulomatous amoebic encephalitis
• From dust, soil
• Transmission by inhalation, ingestion or skin
injury
 Intestinal coccidia
• Cryptosporidium, Cyclospora cayetanensis, Cystoisospora belli, Sarcocystis spp. -gastrointestinal
illness in humans.
• Trophozoite and cystic forms

• Sexual and asexual cycle in single host

• Characterized by fecal excretion of spore like oocysts containing sporozoites, product of sexual
reproduction in the epithelium of the small intestine

• Oocysts are highly infectious

• Live intracellularly part of their lifecycle

• Mode of transmission: contaminated food or water with feces

• Related to poor sanitation and crowded living conditions.

• Usually self limited

• Cause severe protracted diarrheal illnesses in patients with the acquired immunodeficiency
syndrome
 Cryptosporidium
• Cryptosporidium parvum and Cryptosporidium hominis
causes diarrheal disease - cryptosporidiosis.

• Cryptosporidium oocysts in 87% of the raw water samples

• Domesticated animals and birds - primary reservoir

• AIDS patients and children excrete millions of oocysts a
day, survive for months in sewage.

• Spread by direct person-to-person contact.

• The median infectious dose in healthy adult volunteers is
132 oocysts

• Oocysts highly resistant to chlorination.

• The hallmark of cryptosporidial infection is voluminous
• watery diarrhea, explosive and foul smelling with Cryptosporidium parvum life cycle
abdominal cramps, fatigue, and anorexia.
 HIV-infected patients
• Diagnosis:

• Stool Microscopy :Acid-fast staining • Antiretroviral therapy (HAART)

• Supportive measures - antimotility drugs and
• Direct fluorescent antibody [DFA] repletion of electrolyte losses.
• Enzyme immunoassays for detection of • If no resolution of symptoms, Nitazoxanide
Cryptosporidium sp. antigens.
• Molecular methods (e.g., PCR)

• Treatment Complications
 Dehydration- young children, elderly
• Self limiting people-treatable with oral rehydration
therapy.
• Oral rehydration therapy

• Nitazoxanide  In children- linear (height) growth affected

with impaired cognitive function.
• Cryptosporidiosis often not curable and
symptoms return if the immune status
worsens.
 Cryptosporidium
Cystoisospora belli

Cyclospora
Microsporidia
 Flagellates
Giardia (Giardia lambli or Giardia duodenalis)

 Flagellated intestinal protozoan-diarrheal illness giardiasis.

 The infectious form - Cysts

 Trophozoites and cyst forms on surfaces or in soil, food or water contaminated with feces.

 Method of transmission: Water (drinking/recreational water) travelers, person-to-person (daycare

centers),oral anal intercourse

 Diarrhea with flatulence, Foul smelling, frothy, Greasy stools that tend to float, abdominal cramps
nausea/vomiting, dehydration

 In children, delays physical and mental growth, slows development, and cause weight loss and
malnutrition - failure to absorb fat, lactose, vitamin A and vitamin B12.

 Treatment - Metronidazole, Tinidazole, and Nitazoxanide

Life cycle of Giardia

Falling leaf motility

1-10 billion cysts daily in their feces
 Trichomonas vaginalis- Trichomoniasis
 A very common sexually transmitted disease

 A flagellate protozoan parasite –no cystic form

 70% of infected people do not have any signs or symptoms.

 Complain of itching, burning, redness or soreness of the genitals,

discomfort with urination, a frothy, yellowish, or greenish discharge with an
offensive smell

 Microscopy: Wet film preparation with KOH

Treatment a single dose of either

 Metronidazole or Tinidazole
 Features of medically important protozoa
Location Species Mode of transmission Disease
Entamoeba histolytica Amoebiasis
Giardia intestinalis Giardiasis
Cryptosporidium spp. Ingestion of cysts in Cryptosporidiosis
Intestinal tract
Isospora belli food and water Isosporiasis
Cyclospora cayetanensis Cyclosporiasis
Microsporidia Microsporidiosis
Urogenital tract Trichomonas vaginalis Sexual Trichomoniasis
Babesia microti Ixodes scapularis (tick) Babesiosis
Trypanosoma spp. Trypanosomiasis,
Reduviid bug
T. cruzi Chagas disease
Tsetse fly
T. gambiense, T. rhodesiense Sleeping sickness
Leishmania spp. Visceral leishmaniasis
L. donovani complex (kala-azar)
L. tropica, L. major, L. mexicana Sand fly Cutaneous
Blood and tissue L. Viannia braziliensis leishmaniasis
Mucosal leishmaniasis
Plasmodium spp.
P. vivax, P. ovale, P. malariae Anopheles Mosquito Malaria
P. falciparum, P.knowlesi
Toxoplasma gondii Ingestion of cysts in
raw meat; contact
Toxoplasmosis
with soil contaminated
with cat faeces
 HEMOFLAGELLATES
• Live in blood and tissues of human and in gut of insect vectors

• Require two hosts to complete lifecycle

• Family- Trypanosomatidae

• Genus –Trypanosoma and Leishmania

• Exist in 2 or more of the 4 morphological stages:

• Amastigote, Promastigote, Epimastigote, Trypomastigote

• Trypanosoma brucei subspecies - African sleeping sickness

• Trypanosoma brucei rhodesiense (East African sleeping sickness-

eastern and southeastern Africa)

• Trypanosoma brucei gambiense (West African sleeping sickness-

central Africa and in West Africa)

• Transmitted by the tsetse fly (Glossina species)

Distribution of African Trypanosomiasis
 African Trypanosomiasis
• T. b. rhodesiense infection (East African • T. b. gambiense infection (West African
sleeping sickness) sleeping sickness)
• Progresses rapidly • Progresses slowly
• A large sore (chancre) at the site of the tsetse • Itching of the skin, swollen lymph nodes, weight
bite. loss
• Fever, headache, muscle and joint aches, • Intermittent fevers, headaches, muscle and joint
enlarged lymph nodes, HSM within 1-2 weeks aches, and malaise.
of the infective bite.
• After 1-2 years, evidence of central nervous
• After a few weeks, the parasite invades the system involvement, with personality
central nervous system, causes mental changes, daytime sleepiness with nighttime
deterioration and coma. sleep disturbance and progressive confusion.
• Death ensues within months. • Partial paralysis or problems with balance or
walking may occur and hormonal imbalances
noticed.
• Untreated infection lasts 6-7 years, often kills in
Treatment: Pentamidine, Suramin, Melarsoprol
about 3 years.
African Trypanosomiasis

A. Enlargement of the lymph nodes in the neck (Winterbottom's sign)

B. Coma (sleeping sickness) due to generalized encephalitis.
➢ Presently <5 % of Botswana is infested with Tsetse fly
(Ngamiland)

➢ Between 1957-1977 ~ 50 cases/year of trypanosomiasis

were reported in Maun Hospital (Putt, 1984)

➢ No human cases reported since 1984, but 2001 animal

survey showed 16% infection rate
 South American Trypanosomiasis(Chagas disease)

• Trypanosoma cruzi transmitted by reduviid (“kissing”)

bug
• Chancres (“chagomas”) develop at the site of infection,
with transient febrile illness, rarely lead to death by heart
failure.
• Extremely slow and chronic course. Geographic distribution of human
infection with Trypanosoma cruzi
• The parasite invades host cells, macrophages and (Chagas disease).

cardiac muscle cells.

• The major cause of death is myocarditis, progressive
weakening and dilation of the ventricles
• Cardiac aneurysm and heart block are particularly
serious features.
• Dilatation of the intestinal tract -megaoesophagus and
megacolon are the two commonest manifestations Trypanosoma cruzi in the heart muscle of a
child who died of acute Chagas disease. The
infected cardiomyocyte shown contains
several dozen amastigotes
Chagoma is an erythematous and
indurated lesion at the site of parasite
entry that appears a week or two after
transmission has occurred.

When the conjunctiva is the portal of entry,

the patient may develop Romana's sign:
unilateral conjunctivitis, painless palpebral
and periorbital edema and preauricular
lymphadenopathy.
 Leishmania- Leishmaniasis
• Obligate intracellular flagellate protozoa

• Transmitted by sandflies- Phlebotomus papatasi

• Visceral leishmaniasis- Leishmania donovani

• Cutaneous/mucocutaneous leishmaniasis –

• Leishmania tropica, mexicana, and braziliensis,

• Painless small nodules – large ulcers, or plaque

• Heal with scarring

• Immune to reinfection
“wet” or “pizza-like,” with a raised outer border, “dry”,smaller and covered
granulating base, and overlying white, purulent exudates with a crust.
 Phylum Ciliophora (ciliate)
Balantidium coli - Balantidiasis
• Largest protozoan parasite of humans

• Zoonosis: Principal reservoir –pig, monkey, rat

• Trophozoite and cyst form

• Habitat- Large intestine

• Contaminated food and water

• Diarrhoea, dysentery, abdominal colic, tenesmus nausea, vomiting,

mucosal ulcers, abscesses, perforation

• Diagnosis : Microscopy

• Treatment: Tetracycline, Metronidazole

 Toxoplasma gondii-Toxoplasmosis
• Phylum Apicomplexa, coccidian parasite-apical polar complex
• 22.5% of the population 12 years and older have been
infected with Toxoplasma.

• Severe manifestation in immunosuppressed-HIV+

• Not passed from person-to-person

• Definitive hosts - members of family Felidae (domestic
cats and their relatives)

• Exists in three forms:

• Trophozoite (tachyzoites)- acute phase-intracellularly in
tissues, organs

• Tissue cyst (bradyzoites)–chronic phase-brain, muscles

• Oocyst (sporozoites)- definitive host

• Congenital /Acquired Toxoplasmosis

 Lumen dwelling protozoa

http://image.slidesharecdn.com/module12-parasitology-
Blood and tissue dwelling Protozoa

Plasmodium Mosquito Malaria Trophozoites in blood Artemisinin

Babesia microti Ixodes tick Babesiosis Sporozoites Quinine+
Clindamycin
 Thank you!!! Questions???
References:
▪ Mims Medical Microbiology
▪ Mandell, Douglas and Bennett’s Principle and Practice of Infectious
diseases
▪ Tropical Infectious Diseases: Principles, Pathogens and Practice,
Third Edition
 Clinical parasitology; www.medicine.mcgill.ca/tropmed
 http://www.dpd.cdc.gov/dpdx/HTML/Malaria.htm
 http://image.slidesharecdn.com/module12-parasitology
 Parasites without borders- you tube lectures
 https://www.malariasite.com/