DON MARIANO MARCOS MEMORIAL STATE UNIVERSITY

North La Union Campus

Bacnotan, La Union
COLLEGE OF AGRICULTURE

BENEFICIAL ARTHROPODS AND

MICRO-ORGANISM

MODULE 3: NATURE OF BIOLOGICAL

CONTROL AGENTS

(CAMC 103)

JERAMIL C. SIMON
Jay Ron A. Masanit

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 TABLE OF CONTENTS

Lesson Page

1 Parasitoids, Pathogens and Predators

of Insect and Vertebrate 5
Pests . . . . . . . . . . . . . . . . . .

2 Antagonist of Plant 33
Pathogens . . . . . . . . . . . . . . . . . .

3 Biological Control Agents of 43

Weeds . . . . . . . . . . . . . . .

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COURSE OUTLINE

BENEFICIAL ARTHROPODS AND MICRO-ORGANISM

CAMC 103

COURSE DESCRIPTION

The course deals with biology and ecology of beneficial arthropods and micro-
organisms.

COURSE OBJECTIVES

1. Discuss the concepts and principles of biological control of agricultural

plants;
2. Characterize factors affecting the growth and survival of beneficial
arthropods and microorganism;
3. Explain the nature, mechanism and inter action involved among the host
and parasite system; and
4. Outline and discuss the various methods of mass production, application
and evaluation of effectiveness.

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MODULE 3
NATURE OF BIOLOGICAL CONTROL AGENTS

Introduction

A. H. S. Smith (1919) first used term "biological control" to signify the use of
natural enemies (whether introduced or otherwise manipulated) to control
insect pests. Biological control (from the ecological viewpoint) is "the action of
parasites, predators, or pathogens in maintaining another organism's
population density at a lower average than would occur in their absence".

Objectives

After studying this unit, you will be able to:

1. Determine the importance and types of host plant resistance affecting the
pest.
2. Determine parasitoids from predators.
3. Enumerate the different parasitoids, predator’s pathogen, and antagonist
of insect and vertebrate pest.

Lessons

1. Parasitoids, Pathogens and Predators of Insect and Vertebrate Pests

2. Antagonist of Plant Pathogens
3. Biological Control Agents of Weeds

Direction to the Learner

1. Soft copy of this Module will be given/sent to students through on line.

2. Read, understand and follow the instructions given.
3. Answer the questions or activities at the end of the lessons or module.
Write or encode your answers on a bond paper or yellow pad paper.
4. Submit all activities conducted and other requirements on or before the
prescribe schedule. Submission can be done on-line via Facebook
messenger, SMS, or emails.
5. Questions and other queries regarding on the lessons or topics can be sent
or asked through on-line, SMS or during the face to face meetings.
6. Follow the covid-19 protocols during the face to face meetings (if there
is/if necessary). Face to face meeting is not mandatory.

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LESSON 1
PARASITOIDS, PATHOGENS AND PREDATORS

I. Overview

Though most people cringe at the sight of an insect, there are countless
species of beneficial insects. One such group of insects is referred to as natural
enemies. These are insects that prey upon other insects that frequently cause
damage in the garden or landscape. Natural enemies can be divided into
categories: predators, parasitoids, pathogens.

II. Objectives

At the end of this module, the students should be able to;

1. Distinguish the natural enemies of pest as to parasitoid, pathogens, or
predators.
2. Characterized and identify the different natural enemies of pest.
3. Determine the types of pest attacked by a specific natural enemy.

III. Lesson Content

A. Parasitoids

The term parasitoid was coined in 1913 by the German writer O.M. Reuter (and
adopted in English by his reviewer, William Morton Wheeler) to describe the
strategy in which, during its development, the parasite lives in or on the body
of a single host individual, eventually killing that host, the adult parasitoid
being free-living.

Four insect orders that comprise most of the parasitoid

1. Order Hymenoptera 2. Order Diptera
3. Order Strepsiptera 4. Order Coleoptera

1. Parasitoid Hymenopterans (parasitoid wasps)

a. The Importance of Parasitoid Wasps

i. There are many species of parasitoid wasps, but most are so tiny that
they are rarely noticed. What they lack in size they make up in sheer
numbers and efficiency, and as a group they may be the single most
important biological control method gardeners have.

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ii. Wasps belong to the order Hymenoptera, which includes more
parasitoids than any other order of insects, with thousands of parasitic
species in over 40 families.
iii. Parasitoid wasps are very diverse in appearance, ranging in size from as
small as a fleck of pepper up to nearly 3” long, and from uniformly dark
in color to brightly colored and patterned. These tiny agents of death
may be ectoparasitoids or endoparasitoids, but the good news is, they
do not sting people.

b. The Biology of Parasitoids

i. Appearance: Parasitoid wasps are typically so small – most range from

the size of a fleck of pepper to under 1/2” long – that they can only be
reliably identified by an expert.

ii. Life stage(s) that feed on pests: Larvae. Adults usually feed on nectar,
pollen, and honeydew, although a few may feed on host insects as well.

iii. Diet: In general, the eggs, larvae, and sometimes pupae of many
insects, including aphids, caterpillars (larvae of butterflies and moths
Lepidoptera), sawflies, beetles, leafhoppers, true bugs, thrips, psyllids,
and flies.

a) Two species of wasps are very important aphid

parasitoids: Aphelinidae and Aphidius (Braconid wasps).
b) Trichogramma are endoparasitoids of the eggs of over 200 species
of moths and butterflies, and are the most widely released
biological control agents in North America.
c) Encrytids are highly successful generalist feeders that attack a wide
range of host insects.
d) Ichneumons and Braconids are primarily parasitoids of dozens of
different caterpillars (such as armyworms, cabbage looper, fall
webworm, tent caterpillars, tomato fruitworm, redhumped
caterpillar). Cotesia spp. (Braconid wasps) are important
parasitoids of tomato hornworm and imported cabbageworm (see
photo above).

iv. Eggs: Are rarely seen, as they are usually inserted within the eggs or
bodies of host insects.

v. Larvae: Are typically not seen, although some may be glimpsed as a

dark shape within the body or egg of a host insect.

vi. Pupae/Cocoons: The pupae of some parasitoid wasps may be seen as

small whitish/yellowish, rice-like cocoons on or near parasitized
insects.

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 vii. Adults: Parasitoid wasps range in size from very tiny (some can fly
through the eye of a needle) to about 1 ½ inch long. They are not
interested in humans so therefore do not sting.

c. Where to find

Gardeners are more likely to see the results of parasitoids’ activities

than the wasps themselves. Chalcid wasps can be found almost
everywhere, particularly on flowers, foliage, and in leaf litter, but are
rarely noticed because of their tiny size. They may be seen tapping leaf
surfaces with their antennae in search of prey, and they leave sickly or
dead hosts in their wake. Host eggs parasitized by Trichogramma may turn
black as the wasp larva develops within.

d. How to attract and conserve

Parasitoid wasps are very sensitive to insecticides, so avoid or limit the

use of chemical sprays. Most adults feed on plant fluids and sugars, so
provide flowering plants that provide nectar sources. The best nectar
sources are flowers with wide or shallow corollas where the wasps can
easily reach nectar, such as members of the carrot (umbelliferae) and
cabbage (cruciferae) families. Plants with floral nectaries are also
important sources of food, as are aphids and other honeydew producing
sucking insects. Plants that provide shade on hot summer days are a big
help to parasitoids. Trichogramma wasps and those that attack scale
insects, filth flies, aphids, and other insects can be purchased commercially
for release, but it’s important to procure the right species to control the
pest you have.

2. Parasitoid Dipterans

a. Tachinid Fly (Diptera)

i. parasitoid flies are second only to

parasitoid wasps in the sheer
magnitude of pest insects they kill.
ii. There are 12 families of flies with
thousands of species in which some
members are parasitoids, but of
these the tachinids are the most
important.
iii. Most tachinids are endoparasites,
which means that the developing
larvae (maggots) feed within their hosts.
iv. Adult female tachinid flies employ a variety of methods to ensure their
young will have ready access to food as they grow: some lay eggs on

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 leaves to be eaten by caterpillars, others insert eggs or maggots
directly into the host, and still others attach eggs or maggots to the
outside of the host.
v. Eggs consumed by the host or inserted by the mother hatch into
maggots inside the victim. Eggs affixed to the skin of prey hatch and
the maggots bore into the body of the host. Safely inside, the maggots
complete their development, consuming their host as they grow.
vi. Who knew so many flies were doing us so much good in the garden!

b. Life Stage(s) that Feed on Pests

i. Larvae (maggots). Adults usually feed on honeydew, nectar, or pollen,

although a few species feed on host insects as well.

c. Pest(s)/Insects they Feed On

i. Tachinid flies most commonly parasitize the immature life stages (eggs,
larvae or caterpillars, nymphs, and/or pupae) of beetles, butterflies,
and moths, but also earwigs, grasshoppers, sawflies, and true
bugs. Feather legged fly (Trichopoda pennipes) attacks stink bugs and
leaf footed bugs, including squash bug and green stink bug. Istocheta
aldrichi parasitizes adult Japanese beetles.

d. Appearance

i. Eggs: Most tachinids lay small (up to 1/20” in size), oblong, white or
grayish eggs.
ii. Larvae (maggots): usually develop within the host and are not seen.
iii. Pupae: Are commonly small, dark reddish, oblong cases.
iv. Adults: Many resemble house flies in size and color. They have robust
bodies; are usually gray, black, or striped in color; with stout, hairy
bristles protruding from the tip of the abdomen. The Feather legged fly
is bright orange with velvety black head and thorax; with dark legs
(hind legs have a fringe of short, black hairs); yellow feet; large, brown
eyes; and brown and black wings.

e. Where to Find

i. Tachinid flies are found throughout the garden and landscape, and are
frequently mistaken for houseflies.
ii. Feather legged fly is commonly found in the garden laying pale, oval
eggs on the side of squash bugs. Istocheta aldrichi (fly in the family
Tachinidae) may be seen in lawns and shrubbery attaching eggs to the
thorax of newly emerged adult Japanese beetles.

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 iii. The most obvious sign of tachinid fly activity may the presence of
oblong, white eggs glued to the top of the head or body of a host
insect.
f. How to Attract and Conserve

i. Most adult tachinid flies feed on nectar and pollen, especially from
flowering umbelliferous plants such as carrot, dill, and other herbs, and
composite flowers such as asters and rudbeckias , as well as other
flowering plants.
ii. They will also feed on aphid honeydew, so having non-crop plants
infested with aphids helps support tachinid flies.

3. Parasitoid Stripsipterans

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Strepsiptera parasitize four orders of insects: Orthoptera, Hemiptera,
Homoptera and Hymenoptera, with the latter including the largest number of
families. Records of attack on Orthoptera and Hemiptera are few, while both
the Cicadellidae and Fulgoridae frequently are hosts. However, the
Hymenoptera, particularly families Vespidae, Eumenidae and Andrenidae, are
most frequent hosts.

a. Life cycle

In most cases the life cycle is essentially as follows:

i. Males fly about, sniffing around for females which are inside bees or
other host insects.
ii. Then they mate with the female, who doesn't move from the host
insect during this process.
iii. The eggs develop and hatch inside the female's body.
iv. The larvae hatch from
the eggs and walk out
onto the outside of the
host using their six
legs.
v. Then they hail a taxi -
actually, another host
adult, which they grab
onto and which
eventually goes home
to its nest.
vi. It's the nest these larvae are interested in, because there will be larvae
of the host insect there. They disembark from the taxi, pay the fare (I
made that bit up) and enter a larva of the host insect.
vii. Then, they moult into a legless maggot, and live inside it, feeding
away, and mature into an adult shortly after the host eventually
emerges from its pupa.

In some cases strepsipterous larvae are born with legs and then develop
into forms, including the adult female form, that don't have legs (this is
called hypermetamorphosis). After that they just develop by incomplete
metamorphosis.

b. Classification of Strepsiptera

i. In terms of where to place this group among the insects, some

entomologists regard them as somewhat related to beetle groups, on
the basis of similarities among the larvae.

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 ii. The largest families are the Stylopids (Stylopidae) which live on bees
and wasps, and Halictophagidae and Elenchidae, which feed on
planthoppers. The other families are the Mengenillidae, Mengeidae,
Bohartillidae, Corioxenidae, Callipharixenidae and Myrmecolacidae.

Family Hosts
Mengenillidae Unknown
Mengeidae Unknown
Stichotrematidae Orthoptera: Locustidae
Callipharixenidae Hemiptera: Pentatomidae
Myrmecolacidae Hymenoptera: Formicidae
Stylopidae Hymenoptera: various families
Hylecthridae Hymenoptera: Hylaeidae
Xenidae Hymenoptera: various families
Triozoceridae Homoptera: Cicadellidae
Halictophagidae Homoptera: Cicadellidae, Fulgoridae
Elenchidae Homoptera: Cicadellidae, Fulgoridae

c. Where to find them

i. As mentioned above, the males may be seen flying around flowers, but
the easiest way to find these parasites is by collecting some of their
host insects and trying to keep them alive while waiting for the
parasites to emerge. Host insects likely to be parasitised look bloated
and tired. You might spot the female parasite, sticking her head out
from the host's abdomen.

4. Parasitoid Coleopterans

A parasitic life style is not so common in

Coleoptera as it is in Hymenoptera and Diptera,
with ca. 8 families showing this behavior. Most
species of the small family Leptinidae exhibit a
facultative parasitism, and in the Staphylinidae
many species of the Aleocharinae
(Baryodma, Coprochara and Aleochara) are
parasitoids of Diptera puparia. In Cleridae,
several species of Hydnocera are parasitoids,
and some Trichodes seem to develop in the
same way. The Ripiphoridae are entirely
parasitic on hymenopterous larvae and
cockroaches. Some species of Colydiidae
(e.g., Deretaphrus and Bothrideres) are

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considered parasitic, which is true also in Catogenus of the Passandridae. A
few species of Anthribidae of the genus Brachytarsus are considered parasitic
inasmuch as the larval food is strictly limited to the eggs beneath a single
coccid host and the stimulus for oviposition is provided by the scale host itself
rather than by the eggs. A few Coccinellidae, only those which attack the
larger monophlebine Coccidae, may also be thought of as parasitic because the
larva may develop entirely at the expense of a single host individual.

There are few internal parasitoids among Coleoptera, except in the

Ripiphoridae, where it is normal for all species. Among species attacking
cockroaches the entire feeding period is passed internally, while in those
attacking larvae of Hymenoptera, the internal phase is restricted to the latter
portion of the 1st larval instar and the following instars are external
feeders. Internal parasitism by Colydiidae is suspected, especially as isolated
records show larvae being collected from pupae of Chrysobothris.

Parasitic Coleoptera show a considerable uniformity in behavior and the

manner of development. In the families Ripiphoridae, Staphylinidae and
Meloidae, all species deposit their eggs apart from the host stages on which
development is to occur, placing them in the soil, in host galleries, or on
foliage or blossoms. First instar larvae of parasitic Staphylinidae search for
dipterous puparia in the soil of refuse habitat; those of Ripiphoridae attacking
cockroaches, and probably a few Meloidae attacking bees, gain access to the
host directly. The majority of species of the latter two families that attack
vespoid wasps and bees, respectively, seem to require the services of a carrier
to transport them from the vicinity of hatching to the cell, and this role is
usually filled by the female wasp or bee (Clausen 1940/1962). Larval
development among parasitic species also reveals certain points in common
that are not possessed by the predaceous forms. A notable
hypermetamorphosis occurs during larval development. The planidium type of
1st instar larva is of common occurrence in the Meloidae and Ripiphoridae and
in parasitic representatives of several other families. Later larval instars
assume a degenerate form in which the appendages are much reduced and the
powers of locomotion are very limited or entirely lacking. Nonfeeding larval
stages of Meloidae have not been recognized in other parasitic groups of
Coleoptera with the exception of Drilidae (Clausen 1940/1962).

5. General Classification of Parasitoids

Parasitoids live and develop either within their host (endoparasitic) or outside
of it (ectoparasitic). In most endoparasitic species, adult parasitoids oviposit
(lay or insert) an egg into the body of the host, where it grows while gaining
nourishment from within the host’s body. The host dies after the parasitoid
emerges to pupate. In ectoparasitic species, adult parasitoids paralyze the
host and oviposit an egg on the outside of the host’s body. The host is then

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taken back to a nest where the larvae can develop outside of the host’s body
without being disturbed.

a. Endoparasitoids

i. Ichneumonid wasps (Family: Ichneumonidae) differ from paper wasps and

hornets in that they use their stinger to oviposit eggs into a host, rather
than sting for defense. When the egg hatches, the larva feeds on the
innards of its host before eventually boring its way out of the host to
pupate. Common host insects include caterpillars and the larvae or pupae
of beetles, flies, and other wasps. Most ichneumonids are solitary, meaning
a single larva develops and matures within a host. Species vary greatly
from one another, though most are slender wasps with long ovipositors
that are often longer than their bodies. This is the largest wasp family in
North America with over 3,300 species.

ii. Braconid wasps (Family: Braconidae) are generally smaller than

ichneumonids and often go unnoticed. Females oviposit eggs into host
insects similar to ichneumonids and can be solitary or gregarious (many
eggs deposited into a host). The larvae of some species remain attached to
their host after exiting to pupate. This is commonly observed on large
caterpillars, which may have a few dozen pupae attached to their backs.
Other common hosts include aphids, beetles, flies, ants, and bugs.
Braconid species vary greatly in habit and may parasitize the eggs, larva,
pupa, or adult stages of their hosts. Additionally, some species of
braconids are ectoparasitic. This is the second largest wasp family in North
America with 1,900 species.

Insects parasitized with brachonid wasp

iii. Tachinid flies (Family: Tachinidae) are a large family of parasitic flies.
Females of most species lay eggs directly on a host insect. When the egg
hatches, the larva bores its way into the host’s body to feed internally. In
other species, females lay eggs on the surface of a leaf where it may be

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 inadvertently ingested as the host forages. These eggs hatch within the
host’s body. Once fully developed, the larvae exit to pupate in a process
which usually kills the host. Adults are frequently seen foraging flowers,
where they feed on nectar and contribute to pollination.

A leaf footed bug with a tachinid fly egg on its head

b. Ectoparasitoids

i. Sphecid and cabronid wasps (Familys: Sphecidae and Cabronidae) have

long legs and thin waists. Common members of these closely related groups
include mud daubers, thread-waisted wasps, and cicada killers. Most
species nest in the ground, though some, such as the mud dauber,
construct nest out of mud well above the ground. Nests are provisioned
with insects or spiders that have been stung and paralyzed. Eggs are laid
on the prey, which is then fed on as the larvae develop within the nest.
Adults can be observed foraging flowers and feeding on nectar.

Mud dauber nest provisioned with paralyzed spiders inside

B. Predators

Predators may attack their prey as an immature and or adult and require the
consumption of many to survive. There are many different species with various
feeding habits. Some predators eat their prey whole while others suck out the

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bodily fluids of their prey. Many predatory insects are skilled fliers and/or
runners which aids in catching prey. Some predators use intricate crypsis
(camouflage) that allows them to ambush their prey. Others feed on slow,
sedentary insect species; therefore, they do not require speed nor stealth.

Difference Between Predator Specialists and Generalists

Specialists predators feed on only one or a few species of prey, but most are
generalists and feed on a wide variety of insect pests and even, at times, each
other.

i. They may feed on any or all life stages, including eggs, larvae (caterpillars,
grubs, and maggots), nymphs, pupae, or adults.
ii. Some are predators during both their larval and adult stages (e.g., lady
bird beetles), while others are predaceous only in the larval stage (e.g.,
lacewings) and as adults feed on nectar and pollen from flowers.
iii. Predators that eat only other arthropods are called carnivores; arthropods
that eat only plants are referred
iv. to as herbivores and are frequently the prey of predators.
v. Predators that feed on both prey and plants (pollen and nectar) are called
omnivores.

How do predator Insects kill their prey

i. Predators like lady beetles and ground beetles chew and devour their prey.
ii. Others, like assassin bugs, predatory stink bugs, and the larvae of
lacewings and flower flies, have piercing mouthparts and suck the fluids
from the bodies of their prey.
iii. Some are active hunters, stalking and running down their prey; others,
such as dragonflies, may catch dinner on the wing; and still others like
mantises hide patiently in ambush, snatching up unsuspecting victims that
wander too close. However they catch their dinner, predators are a
gardener’s best friend!

List of predatory arthropods

a. Lady beetle/ladybug beetle; b. Green Lacewings, c. Syrphid flies, d.

Predatory bugs, e. Ground beetles, f. Mantis/mantids, g. Hunting wasp, h.
Predatory mites, I. Spiders

1. Lady beetles (Family: Coccinellidae), often called ladybugs, are among

the most recognizable insects and are very common and voracious
predators of aphids. Lady beetles eat aphids whole and may gobble down
as many as 5,000 throughout the course of their life. The adults and larvae
are predacious. Lady beetles also feed on caterpillars, moth eggs, scales,
mites, and they may also be cannibalistic if prey is scarce.

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 Lady bug eggs (yellow mass) laid next to an aphid colony
on the underside of a cotton leaf.

2. Ground beetles and tiger beetles (Family: Carabidae) are common in

almost every environment and come in a variety of shapes, colors, and
sizes. Generally, they have large eyes, long legs, and large, powerful jaws
for grabbing prey. Ground beetle are constantly on the move, searching for
prey by sight, though some species detect prey by scent. Ground beetles
and tiger beetles feed on aphids, caterpillars, other beetle larvae, fly
larvae, mites, springtails, slugs, and even small weed seed. The adults and
larvae are predacious, but larvae live in the soil and have a smaller search
range. Ground beetles can consume up to their body weight each day.

A six-spotted tiger beetle searching

the forest floor for prey

3. Assassin bugs and ambush bugs (Family: Reduviidae) are true bugs,
meaning they have piercing/sucking mouthparts. They feed by inserting
their proboscis (straw-like mouthpart) into their prey, injecting digestive
enzymes, then sucking out the bodily fluids. This is another extremely
diverse group of beneficial insects. Assassin bugs can kill prey that is
significantly larger than they are and are common predators of
caterpillars, aphids, beetles, and others. Ambush bugs use crypsis
(camouflage) to hide on flowers, where they lie in wait for their prey.
When an unsuspecting victim gets too close, they have raptorial (praying
mantid-like) front legs, specialized for grabbing prey. Ambush bugs feed on
flower foraging insects, including small beetles, flies, bees, and wasps.
Adults and nymphs in this family are predacious.

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 An ambush bug lying in wait on An assassin bug sitting on a blooming
goldenrod soybean plant

4. Green lacewings (Family: Chrysopidae) get their name from their lacy-
looking wings. Adults feed on nectar, pollen, and honeydew and are
commonly observed flying around
lights after dark. Eggs are laid at
night on the tip of a thin, hair-like
stalk about a half inch long. This
keeps the eggs up and away from
predators, such as sibling lacewings.
The larvae are wingless; therefore,
they crawl around on plant leaves in
order to search for prey. They are
mainly predators of aphids, but will
also eat small caterpillars or beetle
larvae. Green lacewing larvae can
eat between 100 and 150 aphids in
their lifetime.

5. Syrphid flies (Family: Syrphidae), also called hoverflies, are commonly

observed hovering around flowers, feeding
on nectar and pollen all while contributing
to pollination. Syrphid fly larvae; however,
are voracious predators of aphids, scales,
thrips, and small caterpillars. Adults
frequently lay their eggs among or near
aphid colonies to ensure the larvae have
plenty to eat. Once the blind and legless
larvae hatch, they go to work crawling
around and sucking the bodily fluids out of
their prey. As they visit flowers, adult
syrphid flies are often mistaken for bees or
wasps. An easy way to distinguish them is
to count their wings, assuming they sit still
long enough. Syrphid flies have two wings whereas bees and wasps have
four.

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6. Robber flies (Family: Asilidae) are a highly diverse family of flies. They
typically hunt for prey from a perch
overlooking sunny, open areas. Once located,
robber flies grab their prey in midair and
inject it with neurotoxic saliva which rapidly
immobilizes it. The prey is then carried back
to the perch to be eaten. Enzymes in the
lethal injection help digest the prey’s bodily
fluids, making it easier for the fly to
consume. Males hunt for a mate in a similar
fashion, minus the lethal injection. Robber
flies prey mostly on flying insects such as
other flies, beetles, true bugs, butterflies,
bees, and wasps. Larvae develop in the soil, but are also predacious,
feeding on insect eggs, other larvae, and soft bodied, soil dwelling insects.

7. Dragonflies are one of the oldest groups of winged insects in the world.
There are over 5000 species of dragonflies
worldwide. The largest family,
Libellulidae, includes over 100 species in
North America alone. Dragonflies are
strong, agile fliers and have keen eyesight.
They catch their prey in the air and feed
on almost any insect smaller than they are,
especially midges, mosquitos, butterflies,
and damselflies. Adult females lay eggs by
flying low over the surface of water and
tapping the tail end of their abdomen on
the water, shaking eggs out as they fly.
Eggs may also be laid on aquatic vegetation. Larvae develop in the water
and are predacious on other aquatic insects. Once mature, they climb out
of the water to pupate and take to the skies.

8. Praying Mantids (Family: Mantodea) are easily recognized by their

raptorial front legs that give them
the praying appearance. Crypsis
helps them blend into their
surroundings as they ambush or stalk
prey. Once within reach, they grab
their prey with their front legs in a
lightning fast motion and bite the
victim’s neck to immobilize it.
Mantids will eat nearly anything
they can catch including flies,
moths, crickets, grasshoppers, and even small vertebrates. Females lay
eggs in a dense, foamy case called an ootheca, which contain up to 400

18
 eggs. When the eggs hatch the nymphs may cannibalize each other if no
other food is available.

9. Predatory Mites: Several mite species are predators of plant-feeding

spider mites. Typically, these predatory
mites are a little larger than spider mites
but are more rounded in shape and faster
moving than their prey. Predatory mites
often can provide good control of spider
mites. Low humidity can restrict their
activity. They are also more susceptible to
insecticides than are plant-feeding species.

10. Spiders: All spiders feed on insects or other small arthropods. Most people
are familiar with many common web-making
species. However, there are many other
spiders — wolf spiders, crab spiders,
jumping spiders — that do not build webs
but instead move about and hunt their prey
on soil or plants. These less conspicuous
spiders can be important in controlling
insect pests such as beetles, caterpillars,
leafhoppers and aphids.

C. Pathogen of Insects

Pathogens are viruses or microorganisms that cause disease. Like all other
organisms, insects are susceptible to a variety of diseases caused by
pathogens. Many of these pathogens cause disease that are acute and fatal and
therefore are used as model to study processes of infection and pathogenesis
as well as to control populations of insects that are pests or vectors of plant
and animals diseases.

Four main groups of Insect Pathogens

1. Viruses

Viruses are obligate intracellular parasites, meaning that they can reproduce
only in living cells and are composed in the simplest form of a nucleic acid,
either DNA or RNA, and a protein shell referred to as the capsid. More complex
viruses also contain a lipoprotein envelop. Insect viruses can be cultured in
living hosts (in vivo) or in cultures insect cells (in vitro).
i. Iridoviruses
ii. Cytoplasmic Polyhedrosis Viruses
iii. Entomopoxviruses

19
iv. Ascoviruses
v. Baculoviruses
vi. Nuclear Polyhedrosis Virus
vii. Granulosis Virus

a. Invertebrate iridescent viruses (Iridoviridae)

Invertebrate iridescent viruses

(IIVs) are icosahedral particles of
120 – 180 nm in diameter,
containing an internal lipid
membrane and a double-stranded
DNA genome of 130 – 210 kbp,
that belong to the family
Iridoviridae. They mostly infect
arthropods, particularly insects,
in damp or aquatic habitats worldwide. IIV particles are organized into
crystalline arrays. Light reflected from such arrays interferes with incident
light, resulting in the characteristic iridescent colours that are the most
obvious sign of patent infection. Patent disease is lethal in the larval or
pupal stages.

The small IIVs (genus Iridovirus) tend to display colours from violet to
turquoise. The large IIVs, that infect mosquito and midge larvae (genus
Chloriridovirus), tend to display colours from green to orange or red.

b. Cytoplasmic Polyhydrosis Virus

The cytoplasmic polyhedrosis viruses

(family Reoviridae) are occluded double-
stranded RNA viruses with a genome
divided into 9 or 10 segments of RNA.
These viruses, commonly referred to as
CPVs, cause a chronic disease and
reproduce only in the stomach of insects,
where typically they form large (ca. 0.5–2
μm) polyhedral to spherical occlusion
bodies in the cytoplasm of midgut
epithelial cells. Infection in early instars
retards growth and development,
extending the larval phase by weeks. The
disease is often fatal. In advanced stages of disease, the infected midgut is
white rather than translucent brown, because of large numbers of
accumulated polyhedra. This virus type is relatively common among
lepidopterous insects and among dipterous insects of the suborder
Nematocera (e.g., mosquitoes, blackflies, midges). CPVs are typically easy

20
to transmit by feeding to species that belong to the same family of the host
from which they were isolated, and thus the host range of this virus type is
quite broad.

c. Entomopoxviruses

The entomopoxviruses (family Poxviridae)

are occluded double-stranded DNA viruses
that produce large, enveloped virions (150
nm × 300 nm) that replicate in the
cytoplasm of a wide range of tissues in
most hosts, causing an acute, fatal disease.
Occlusion bodies vary from being oval to
spindle shaped and generally occlude 100
or more virions. These viruses have been
most commonly reported from
coleopterans, from which there are over 30
isolates, but they are also known from
lepidopterous, dipterous (midges), and
orthopterous (grasshoppers) insects. This
virus type is easily transmitted by feeding, although where the
experimental host range of individual isolates has been tested, it has been
found to be relatively narrow, generally being restricted to closely related
species. Insect poxviruses are related to vertebrate poxviruses, such as the
variola virus, the etiological agent of smallpox, and they may be the
evolutionary source of the vertebrate poxviruses.

d. Ascoviruses

The ascoviruses (Ascoviridae) are a

new family of DNA viruses, at present
known only from larvae of species in
the lepidopteran family Noctuidae,
where they have been reported from
several common pest species such as
the cabbage looper, cotton
budworm, corn earworm, and fall
armyworm. Ascoviruses cause a
chronic, fatal disease of larvae. The
virions of ascoviruses are large (130
nm × 400 nm), enveloped, and
reniform to bacilliform in shape; they exhibit complex symmetry and
contain a circular, double-stranded DNA genome. During the course of
ascovirus disease, large numbers of virion-containing vesicles accumulate in
the blood of infected caterpillars, changing its color from translucent green
to milky white. These virion-containing vesicles are formed by a unique

21
developmental sequence resembling apoptosis (cell death) in which each
infected host cell cleaves into a cluster of vesicles as virion assembly
proceeds.

An interesting ascovirus feature is that transmission from host to host

depends on vectoring by female endoparasitic wasps. Ascoviruses are very
difficult to transmit by feeding, with typical infection rates averaging less
than 15% even when larvae are fed thousands of vesicles in a single dose. In
contrast, infection rates for caterpillars injected with as few as 10 virion-
containing vesicles are typically greater than 90%, and experiments with
parasitic wasps show that these insects can transmit ascoviruses.

e. Baculoviruses

The baculoviruses (family: Baculoviridae) are a group of large DNA viruses

that infect insects. These viruses are well known for their utility and
versatility as gene expression vectors, biological pesticides, and vectors for
transduction of mammalian cells.

A typical baculovirus replication cycle: Nucleocapsids are produced in the

nucleus of infected cells, but two forms of enveloped virions are produced.
The first, budded virus (BV), is formed when single nucleocapsids exit the
nucleus and bud from the cell, acquiring an envelope from the plasma
membrane. The BV attachment and fusion protein, either GP64 or F
depending on the type of baculovirus, is concentrated at one end of the
virion in structures that are visible by electron microscopy, known as
peplomers. Later, the second type of virion, called occlusion-derived virus
(ODV), is formed when nucleocapsids obtain an envelope from the inner
nuclear membrane. ODV become embedded in large proteinaceous crystals
that are primarily composed of a single viral protein called polyhedrin, and

22
are known as occlusion bodies. The occlusion bodies remain within the
nucleus until they are liberated by cell lysis. The ODV of some
baculoviruses, known as multiple nucleopolyhedroviruses (MNPVs), contain
multiple nucleocapsids within a single-enveloped virus particle, as is
illustrated here. In nature, the baculovirus replication cycle begins when a
susceptible insect larva consumes viral occlusion bodies contaminating their
food source. The occlusion bodies dissolve in the highly alkaline
environment of the larval midgut, releasing ODV, which attach to the
microvillar membranes of midgut epithelial cells. Attachment and fusion
occurs via the PIF proteins, found in the ODV envelope. Infected midgut
epithelial cells produce BV, which bud from the basal side and infect
tracheal epithelial cells. Infection of tracheal cells is thought to be one
mechanism that allows BV to escape across the midgut basal lamina (BL)
and spread infection throughout the insect. The majority of tissues become
infected, producing large amounts of ODV and BV. The infected insects
liquefy after death, allowing dispersal of occlusion bodies and promoting
subsequent infections.

f. Nuclear Polyhedrosis Virus

The nuclear polyhedrosis virus (NPV),

part of the family of baculoviruses, is a
virus affecting insects, predominantly
moths and butterflies. It has been used
as a pesticide. The polyhedral capsid
from which the virus gets its name is an
extremely stable protein crystal that
protects the virus in the external
environment. It dissolves in the alkaline midgut of moths and butterflies to
release the virus particle and infect the larva. An example of an insect that
it infects is the fall webworm.

Symptoms of NPV infection include: Discoloration (brown and yellow),

Stress (regurgitation), Decomposition (liquefication), Lethargy (slow
movement to no movement at all; refusal to eat).

The virus enters the nucleus of infected cells and reproduces until the cell
begins to produce crystals in the fluids of the host. These crystals can
transmit the virus from one host to another. The host becomes visibly
swollen with fluid containing the virus, and eventually dies, turning black
with decay. Mortality in infected insects is nearly 100%.

g. Granulosis Virus

Granuloviruses are in the family of insect viruses called the baculoviridae.

These viruses target insects and are popular as insecticides for farmers

23
 targeting lepidopteran larva. The viruses
are present in many historical records for
causing silkworm 'jaundice' and have
been used as biopesticides since WWII.
There are currently 17 species in the
genus betabaculovirus. These viruses are
popular due to their selectivity of only
attacking insects from the order
lepidotera. Granuloviruses are well
known for their unique ability to
completely liquefy their hosts in order to spread to more hosts, a trait they
share with the closely related nuclear polyhedrosis viruses. The type
species of granuloviruses is the species Cydia pomonella granulosis
virus (CpGV), which only affects Cydia pomonella larva. C. pomonella larva
are pests of various fruits in agriculture, causing great loss of ripened
fruits. Granulovirus has been used as a pesticide since World War II
(Federici, 1997); however, C. pomonella has been developing resistance to
granulovirus since 2005 (Sauer, 2017). Granuloviruses are useful in the
pesticide industry due to their ability to efficiently kill lepidopteran pests,
such as C. pomonella, to protect crops without damaging them or harming
the consumer of the crops. Extensive research is being done on more uses
and applications of baculoviridae to operate as pesticides against more
specialized or resistant lepidopteran pests.

2. Bacteria

Bacteria ate relatively simple unicellular microorganisms that lack internal

organelles such as a nucleus and mitochondria, and reproduce by binary
fission. With a few exceptions, most of those that cause disease in insect grow
readily on a wide variety of inexpensive substrates, a characteristics that
greatly facilitates their mass production. A variety of bacteria are capable of
causing diseases in insects, but those that have received the most study are
spore-forming bacilli (family Bacillaceae), especially B. thuringiensis.
i. Bacillus thuringiensis
ii. Bacillus sphaericus
iii. Paenibacillus popilliae
iv. Serratia entomophila

a. Bacillus thuringiensis (Bt)

Bt is a soil-dwelling bacterium that naturally

produces a toxin that is fatal to certain
herbivorous insects. The toxin produced by
Bacillus thuringiensis (Bt) has been used as
an insecticide spray since the 1920s and is

24
commonly used in organic farming. Bt is also the source of the genes used
to genetically modify a number of food crops so that they produce the toxin
on their own to deter various insect pests. The toxin is lethal to several
orders of insects, including Lepidoptera (butterflies, moths, and skippers),
Diptera (flies), and Coleoptera (beetles), though a number of Bt strains are
available to make its use more target-specific.

Application: Bt was first discovered in 1901 by a Japanese scientist

investigating the decline in silkworm moth populations, which he attributed
to the rod-shaped, Gram-positive bacterium. In 1911 it was rediscovered by
a German scientist, and a solution of crystallized Bt toxins was found to be
highly effective against certain crop pests, including the corn borer, corn
rootworm, corn earworm, and bollworms. In the United States the product
was first used commercially as an insecticide spray in 1958, and several
different strains of the bacterium are currently used to control for a
number of agricultural insect pests and their larvae. Bt toxin can be applied
to crops, including potatoes, corn, and cotton, as a spray or, less
commonly, in granular form.

Bt proteins can also be introduced into the crops themselves through

genetic engineering. Bt crop varieties are engineered to produce a protein
toxic to specific insects and are used in areas with high levels of
infestations of the targeted pests. Since 1995, when the U.S. Environmental
Protection Agency (EPA) first approved use of the technology, commercial
production of Bt corn, cotton, potatoes, and rice has increased
dramatically in many countries, though plantings often fluctuate depending
on pest infestation levels.

Properties: Susceptible insects must ingest Bt toxin crystals in order to be

affected. In contrast to poisonous insecticides that target the nervous
system, Bt acts by producing a protein that blocks the digestive system of
the insect, effectively starving it. Bt is a fast-acting insecticide: an
infected insect will stop feeding within hours of ingestion and will die,
generally from starvation or a rupture of the digestive system, within days.

Whether applied in spray form or through genetic engineering, each Bt

strain is effective against a narrow range of insects. The most commonly
used strain of Bt (kurstaki, or Btk) targets only certain species of
caterpillars. Since the late 1970s, Bt strains (e.g., israelensis, or Bti) have
been developed that control certain types of fly larvae, including those of
mosquitoes, black flies, and fungus gnats. Other common strains include
san diego and tenebrionis, which are effective against certain leaf beetles,
such as the Colorado potato beetle and elm leaf beetle.

b. Bacillus sphaericus

25
Lysinibacillus sphaericus (reclassified - previously known as Bacillus
sphaericus) is a Gram-positive, mesophilic, rod-shaped bacterium
commonly found on soil. It can form resistant endospores that are tolerant
to high temperatures, chemicals
and ultraviolet light and can remain viable
for long periods of time. It is of particular
interest to the World Health
Organization due to the larvicide effect of
some strains against two mosquito genera
(Culex and Anopheles), more effective
than Bacillus thuringiensis, frequently
used as a biological pest control. L.
sphaericus cells in a vegetative state are
also effective against Aedes
aegypti larvae[3], an important vector
of yellow fever and dengue viruses.

The high toxicity strains produce during sporulation a

binary toxin composed of BinA (42 kDa) and BinB (51 kDa) proteins, which is
the major insecticidal component. The protein BinB acts by binding to
a receptor in the epithelial midgut cells, facilitating the entrance
of BinA which causes cellular lysis. After being ingested by larvae, these
proteins are solubilized in the gut and undergo proteolysis to active
lower molecular weight derivatives. The vegetative cells of both high- and
low-toxicity strains produce Mtx1, Mtx2 and Mtx3 toxins,
but Mtx1 and Mtx2 are degraded by proteases during the stationary phase,
consequently making them undetectable in sporulated cultures. In addition,
the presence of binary-toxin genes and proteins has been determined in 18
pathogenic strains. Strains OT4b.2, OT4b.20, OT4b.25, OT4b.26 and
OT4b.58 were found as toxic as the spores of the reference WHO strain
2362, against C. quinquefasciatus larvae.

c. Paenibacillus popilliae

Milky Disease (Eubacteriales: Bacillaceae) formerly Bacillus popilliae

Japanese beetle is the exclusive host of

the strain of P. popilliae which is sold
commercially. However, other P.
popilliae strains (and P. lentimorbus,
which is considered a strain of P.
popilliae by some experts) have other
scarab hosts and are specific to different
beetles in the family Scarabaeidae,

26
 which includes the Japanese beetle and the chafers - important pasture
pests, but also the beneficial dung beetles.

Spores which reside in the soil and have been ingested by beetle larvae
germinate in the larva's gut within 2 days and the vegetative cells
proliferate, attaining maximum numbers within 3 to 5 days. By this time,
some of the cells have penetrated the gut wall and have begun to grow in
the hemolymph, where large numbers of cells develop by day 5 to 10. A
few spores also are formed at this stage, but the main phase of sporulation
occurs later and is completed by 14 to 21 days when the larva develops the
typical milky appearance.

In laboratory conditions, the larva remains alive until this stage and usually
contains about 5 x 109 spores. In field conditions, however, there are
reports that larvae sometimes die earlier, before the main phase of
sporulation is completed. This is of concern because sporulation stops when
the host dies and the larva ultimately releases fewer spores to maintain the
level of infestation of a site.

d. Serratia entomophila

A novel bacterium named Serratia

entomophila causes amber disease in the
grass grub, Costelystra zealandica, an
important pest of pastures in New Zealand,
and has been developed as a biological
control agent for this pest (Jackson et
al., 1992). This bacterium adheres to the
chitinous initma of the foregut, were it grows
extensively, eventually causing the larvae to
develop an amber color and resulting in
death. The bacterium is easily grown and
mass-produced in vitro, and can now be grown to densities as high as 4 ×
1010 cell per milliliter. Successful mass production of S. entomophila led to
its rapid commercialization. It is now used to treat infested pastures in
New Zealand at a rate of 1 liter of product per hectare. Liquid formulations
of this living non-spore-forming bacterium are applied with subsurface
application equipment. The rapid development and commercialization of
the bacterium, even though the use is rather restricted, shows how
microbials can be successful in niche markets where there are few
alternatives, and where mass production methods—the most critical factor
—are available.

3. Fungi

27
Unlike most other pathogens, fungi usually infect insects by active penetration
through the cuticle. The typical life cycle begins when a spore, either a motile
spore or a conidium, lands on the cuticle of an insect. Soon after, under
suitable conditions, the spore germinates, producing a germ tube that grows
and penetrates down through the cuticle into the homocoel. Once in the
hemolymph, the fungus colonizes the insect. Hyphal bodies bud off from the
penetrate hyphae that grow throughout the insect body. Complete colonization
of the body typically requires 7-10 days, after which the insect dies.
i. Aquatic fungi
ii. Terrestrial fungi
iii. Entomophthorales
iv. Class Hyphomycetes

a. Beauveria bassiana

Beauveria bassiana is a
fungus which causes a
disease known as the
white muscadine
disease in insects.
When spores of this
fungus come in contact
with the cuticle (skin)
of susceptible insects, they germinate and grow directly through the
cuticle to the inner body of their host. Here the fungus proliferates
throughout the insect's body, producing toxins and draining the insect of
nutrients, eventually killing it. Therefore, unlike bacterial and viral
pathogens of insects, Beauveria and other fungal pathogens infect the
insect with contact and do not need to be consumed by their host to cause
infection. Once the fungus has killed its host, it grows back out through the
softer portions of the cuticle, covering the insect with a layer of white
mold (hence the name white muscadine disease). This downy mold
produces millions of new infective spores that are released to the
environment.

Beauveria is a naturally occurring fungus in soils throughout the northeast

(and the world!) and has been researched for control of soil borne insects
(e.g. the May beetle in Europe, the Argentine stem weevil in New Zealand).
Many soil insects, however, may have a natural tolerance to this pathogen,
which is not exhibited in many foliar pests. Therefore, commercial
development of this fungus for biological control has primarily been
targeted against foliar feeding pest.

b. Metarhizium anisopliae

28
Infections of arthropods by Metarhizium species are easily recognized a
few days after death, when the fungus grows out of the arthropod
integument and forms reproductive structures. Initially, one only sees
fungal hyphae that appear white, but, as conidia form and mature they
often take on a characteristic olive green color. However, depending on
the species and strain of Metarhizium, spores can range in color from white
to yellow to brown and green (Tanada and Kaya 1993).

Generally, the development of a lethal arthropod infection can be

separated into three stages. First, asexual conidia (singular=conidium)
come into contact with the arthropod integument as the arthropod travels
through the environment. The conidia stick to the arthropod’s exoskeleton,
germinate and grow a germ tube, which eventually ends in an
appressorium, the flattened and thickened tip of a germ tube. A
penetration peg grows under the appressorium, pierces the integument and
enters the hemocoel. The penetration of the fungus is achieved by the
production of a cocktail of hydrolytic enzymes including proteases, lipases,
chitinases, and mechanical pressure. Second, single cells of the fungus,
blastospores, bud off of the penetration structure, circulate in the insect
hemocoel and multiply, thereby depleting host
nutrients. Metarhizium species are also known to produce compounds that
are toxic to arthropods and presumably aid in killing the host, suppressing
host immune defenses and fending off potential microbial competitors.
Finally, after the host dies due to mycosis, the fungus will penetrate out of
the integument and grow conidiophores, on which environmentally stable
aerial conidia are produced. These conidia are passively disseminated into
the environment and eventually infect new hosts.

c. Isaria fumosorosea

Isaria fumosorosea is an
entomopathogenic fungus, formerly
known as Paecilomyces fumosoroseus.
It shows promise as a biological
pesticide with an extensive host range.
This fungus has a wide host range that
includes insects in over twenty five
different families and many species of
mite. Agricultural pest insects which
are susceptible to infection include
the diamondback moth (Plutella
xyllostella), the Russian wheat
aphid (Diuraphis noxia) and
the silverleaf whitefly (Bemisia argentifolii). Among mites, susceptible
species include the spotted spider mite (Tetranychus urticae),

29
 the European red mite (Panonychus ulmi), the brown mite (Byrobia
rubrioculus) and the apple rust mite (Aculus schlectendali).

When a conidium or blastospore of Isaria fumosorosea lands on a suitable

host, it produces enzymes to penetrate the insect's cuticle. A germ tube
then grows into the haemocoel and the fungus proliferates inside the
insect’s body. The fungus can also enter through the spiracles,
the mouth or the anal opening. The mycelia spread in the haemolymph and
tissues, eventually emerging from the insect and producing conidia.
Mortality of the insect has been ascribed to the drainage of its nutrients,
the destruction of its tissues and the release of toxins.

4. Protozoa and Microsporidia

Protozoa is a general term applied to a large and diverse group of eukaryotic

unicellular motile microorganisms that belong to what is now known as the
kingdom Protista. Those that are parasitic have the general feature of causing
diseases that are chronic. Many of the parasitic types especially the
microsporidia, build up slowly in insect populations, eventually causing
epizootics that lead to rapid declines in populations of specific species. These
epizootics attracted interest in the possibility of using protozoa to control pest
insects, and over the past several decades numerous studies have been aimed
at evaluating this potential. E.g. Microsporidia

Mode of Action

Most microsporidia must be eaten to infect an insect, but there may also be
some natural transmission within a pest population, for example by predators
and parasitoids. The pathogen enters the insect body via the gut wall, spreads
to various tissues and organs, and multiplies, sometimes causing tissue
breakdown and septicemia.

Symptoms

Infected insects may be sluggish and smaller than normal, sometimes with
reduced feeding and reproduction, and difficulty molting. Death may follow if
the level of infection is high. One advantage of this type of infection is that
the weakened insects are more likely to be susceptible to adverse weather and
other mortality factors.

Some Beneficial Microsporidia

1. Nosema pyrausta (=Perezia pyraustae) is a microsporidium that infects

several insect species, including European corn borer, for which it can be
an important natural control. This disease was widespread in the

30
 midwestern United States during the 1950s and '60s, causing considerable
natural mortality, but its commercial use is still in the developmental
phase. Infection can spread from diseased to healthy larvae via
contaminated frass, and by migration of infected larvae between plants.
2. Nosema locustae is the only commercially available species of
microsporidium, marketed under several labels for the control of
grasshoppers and crickets. It is applied with an insect-attractant bait.
Because of its slow mode of action, this product is better suited to long-
term management of rangeland pests than to the more intensive demands
of commercial crop or even home garden production.
Other Nosema species have been shown to infect spider mites and
webworms, but have yet to be developed sufficiently for commercial use.
3. Vairimorpha necatrix is another microsporidium with commercial
potential. It has a wide host range among caterpillar pests, including corn
earworm and European corn borer, various armyworms, fall webworm, and
cabbage looper. It can be more virulent than other species and infected
insects may die within six days of infection.

IV. Evaluation (Self-check)

1. Differentiate the following organisms: (15 points)

a) Parasitoid
b) Predator
c) Entomopathogens

2. Briefly discuss the mode of action of the following organisms in infecting

insects:
a) Fungi (15 points)
i. Beauveria bassiana
ii. Metarhizium anisopliae
iii. Isaria fumosorosea
b) Bacteria (20points)
i. Bacillus thuringiensis
ii. Bacillus sphaericus
iii. Paenibacillus popilliae
iv. Serratia entomophila
c) Virus (35 points)
i. Invertebrate iridescent viruses (Iridoviridae)
ii. Cytoplasmic Polyhydrosis Virus
iii. Entomopoxviruses
iv. Ascoviruses
v. Baculoviruses
vi. Nuclear Polyhedrosis Virus
vii. Granulosis Virus

31
V. Home-based Laboratory Activity

1. Collection and preservation of predators and parasitoids

a) Collect available predators and parasitoids of insects and other
arthropods in your surrounding.
b) Preserve your collection using 75% alcohol in a vial. Label your
collections by giving the: a) common name; b) scientific name; c)
family name; d) order; e) name of collector.
c) Place your collection in a box (shoe box)/ plastic box that could
accommodate your collections.
d) A minimum of 50 specimens (collection) is needed.
e) Document all activities conducted and make a report paper to be
passed later. The format will be:
i. Title
ii. Introduction
iii. Materials
iv. Methodology/procedures
f) Write or encode your report on a short bond paper.
i. For type written, use: Arial fon style; size 12; single spacing; and
normal margin.
ii. Write/ encode your name, subject, year and section on the top of
your paper.

VI. Project

1. Biocon agent collection:

a) Minimum of 50 individual biocon agents preserved in a vial with 75%
alcohol.
b) Properly labeled
c) Place your collection in a container (boxes)
d) Present your collection before final exam

2. Compendium of collected biocon agents

a) Make a compendium of your collection. The compendium should be
composed of the following:
i. Photo of the collected individual specimens (minimum of 2x2”).
ii. Names: common name, scientific name, family name, order.
iii. Biology/life cycle
iv. Distribution
v. Host range or prey
b) Use short bond paper following the format of:
i. Margin - normal
ii. Font - arial;11
iii. Spacing - single
c) Present your compendium on or before the final exam.

32
33
LESSON 2
ANTAGONIST OF PLANT PATHOGENS

I. Overview

Plant diseases need to be controlled to maintain the quality and abundance of

food, feed, and fiber produced by growers around the world. Different
approaches may be used to prevent, mitigate or control plant diseases. Beyond
good agronomic and horticultural practices, growers often rely heavily on
chemical fertilizers and pesticides. Such inputs to agriculture have contributed
significantly to the spectacular improvements in crop productivity and quality
over the past 100 years. However, the environmental pollution caused by
excessive use and misuse of agrochemicals, as well as fear-mongering by some
opponents of pesticides, has led to considerable changes in people’s attitudes
towards the use of pesticides in agriculture. Today, there are strict regulations
on chemical pesticide use, and there is political pressure to remove the most
hazardous chemicals from the market. Additionally, the spread of plant
diseases in natural ecosystems may preclude successful application of
chemicals, because of the scale to which such applications might have to be
applied. Consequently, some pest management researchers have focused their
efforts on developing alternative inputs to synthetic chemicals for controlling
pests and diseases. Among these alternatives are those referred to as
biological controls.

II. Objectives

At the end of this lesson, the students should be able to:

1. Determine the different mechanisms of biological control for plant
diseases.
2. Discuss the mode of action for every plant disease antagonists.

III. Lesson Content

A. Types of interactions contributing to biological control

Throughout their lifecycle, plants and pathogens interact with a wide variety
of organisms. These interactions can significantly affect plant health in various
ways. In order to understand the mechanisms of biological control, it is helpful
to appreciate the different ways that organisms interact. Note, too, that in
order to interact, organisms must have some form of direct or indirect contact.
Odum (1953) proposed that the interactions of two populations be defined by
the outcomes for each. The types of interactions were referred to as
mutualism, protocooperation, commensalism, neutralism, competition,

34
amensalism, parasitism, and predation. While the terminology was developed
for macroecology, examples of all of these types of interactions can be found
in the natural world at both the macroscopic and microscopic level. And,
because the development of plant diseases involves both plants and microbes,
the interactions that lead to biological control take place at multiple levels of
scale.

1. Amensalism (Antibiosis and Lysis)

Amensalism is a phenomenon where one population adversely affects the
growth of another population whilst itself being unaffected by the other
population. Generally amensalism is accomplished by secretion of
inhibitory substances.

2. Competition
Among microorganisms competition exists for nutrients, including oxygen
and space but not for water potential, temperature or Ph. Amensalism
involves the combined action of certain chemicals such as toxins,
antibiotics and lytic enzymes. Success in competition for substrate by any
particular fungal species is determined by competitive saprophytic ability
(Garrett, 1950) and inoculums potential (Garrett, 1956) of that species.
Competitive saprophytic ability is “the summation of the physiological
characteristics that make for success in competitive colonization of dead
organic substrates” (Garrett, 1956). Garrett (1950) has suggested four
characteristics which are likely to contribute to the competitive
saprophytic ability:
i. rapid germination of fungal propagules and fast growth of young
hyphae towards a source of soluble nutrients,
ii. appropriate enzyme equipment for degradation of carbon
constituents of plant tissues,
iii. excretion of fungistatic and bacteriostatic growth products
including antibiotics, and
iv. olerance of fungistatic substances produced by competitive
microorganisms.

3. Predation and Parasitism

Predation is an apparent mode of antagonism where a living microorganism
is mechanically attacked by the other with the consequences of death of
the farmer. It is often violent and destructive relationship. Parasitism is a
phenomenon where one organism consumes another organism, often in
asubtle, non-debilitating relationship. These aspects are dealt with the
example of fungi, nematodes and amoebae.

4. Mutualism
In mutualistic interactions, both species benefit from the interaction. A
classic example of mutualism is the relationship between insects that
pollinate plants and the plants that provide those insects with nectar or

35
 pollen. Another classic example is the behavior of mutualistic bacteria in
ecology and human health. Gut bacteria in particular are very important
for digestion in humans and other species. In humans, gut bacteria assist in
breaking down additional carbohydrates, out-competing harmful bacteria,
and producing hormones to direct fat storage. Humans lacking healthy
mutualistic gut flora can suffer a variety of diseases, such as irritable
bowel syndrome. Some ruminant animals, like cows or deer, rely on special
mutualistic bacteria to help them break down the tough cellulose in the
plants they eat. In return, the bacteria get a steady supply of food.

5. Protocooperation
Protocooperation is where two species interact with each other
beneficially; they have no need to interact with each other - they interact
purely for the gain that they receive from doing this. It is not at all
necessary for protocooperation to occur; growth and survival is possible in
the absence of the interaction. The interaction that occurs can be between
different kingdoms.

The term, initially used for intraspecific interactions, was popularized by

Eugene Odum (1953), although other authors prefer to use the terms
"cooperation" or "mutualism".

6. Commensalism
Commensalism is a type of relationship between two living organisms in
which one organism benefits from the other without harming it. A
commensal species benefits from another species by obtaining locomotion,
shelter, food, or support from the host species, which (for the most part)
neither benefits nor is harmed. Commensalism ranges from brief
interactions between species to life-long symbiosis.

7. Neutralism
Neutralism (a term introduced by Eugene Odum) describes the relationship
between two species that interact but do not affect each other. Examples
of true neutralism are virtually impossible to prove; the term is in practice
used to describe situations where interactions are negligible or
insignificant.

8. Other Interactions

a) Mycoparasitism: When one fungus is parasitized by another one the

phenomenon is called as mycoparasitism. The parasitizing fungus is
called hyperparasite and the parasitized fungus as hypoparasite.
Mycoparasitism commonly occurs in nature. As a result of inter-fungus
interaction i.e. fungus-fungus interaction, several events take place
which lead to predation viz., coiling, penetration, branching,
sporulation, resting body production, barrier formation and lysis.

36
 b) Nematophagy: This is the phenomenon of eating upon nematodes by
fungi. However, several nematode eating i.e. nematophagous fungi
(NF) are known which develop different kinds of trap (T), arrest the
pathogenic nematodes (N) and finally kill them.

c) Mycophagy: Mycophagy is the phenomenon of feeding on fungi by

amoebae. In recent years, mycophagy has become a new field of
research as far as biocontrol of soil-borne plant pathogens is
concerned. Many soil amoebae are known to feed on pathogenic fungi.
For example, take-all disease of wheat caused by Gaeumannomyces
graminis var. triticiwas very severe in Australia. Finally it was found
that some natural soil exhibited suppressiveness against this disease.

B. Mechanisms of Biological Control

Types of interspecies antagonisms leading to biological control of plant

pathogens.
Type Mechanism Examples
Lytic/some nonlytic mycoviruses
Ampelomyces quisqualis
Hyperparasitism/
Direct antagonism Lysobacter enzymogenes
predation
Pasteuria penetrans
Trichoderma virens
2,4-diacetylphloroglucinol
Mixed-path
Antibiotics Phenazines
antagonism
Cyclic lipopeptides
Chitinases
Lytic enzymes Glucanases
Proteases
Ammonia
Unregulated waste
Carbon dioxide
products
Hydrogen cyanide
Blockage of soil pores
Physical/chemical
Germination signals consumption
interference
Molecular cross-talk confused
Exudates/leachates consumption
Indirect
Competition Siderophore scavenging
antagonism
Physical niche occupation
Contact with fungal cell walls
Induction of host Detection of pathogen-associated,
resistance molecular patterns
Phytohormone-mediated induction

37
1. Hyperparasites and predation

In hyperparasitism, the pathogen is directly attacked by a specific BCA that

kills it or its propagules. In general, there are four major classes of
hyperparasites: obligate bacterial pathogens, hypoviruses, facultative
parasites, and predators. Pasteuria penetrans is an obligate bacterial pathogen
of root-knot nematodes that has been used as a BCA. Hypoviruses are
hyperparasites. A classical example is the virus that infects Cryphonectria
parasitica, a fungus causing chestnut blight, which causes hypovirulence, a
reduction in disease-producing capacity of the pathogen. The phenomenon has
controlled the chestnut blight in many places (Milgroom and Cortesi 2004).
However, the interaction of virus, fungus, tree, and environment determines
the success or failure of hypovirulence. There are several fungal parasites of
plant pathogens, including those that attack sclerotia (e.g. Coniothyrium
minitans) while others attack living hyphae (e.g. Pythium oligandrum). And, a
single fungal pathogen can be attacked by multiple hyperparasites. For
example, Acremonium alternatum, Acrodontium crateriforme, Ampelomyces
quisqualis, Cladosporium oxysporum, and Gliocladium virens are just a few of
the fungi that have the capacity to parasitize powdery mildew pathogens (Kiss
2003). Other hyperparasites attack plant-pathogenic nematodes during
different stages of their life cycles (e.g. Paecilomyces lilacinus and Dactylella
oviparasitica). In contrast to hyperparasitism, microbial predation is more
general and pathogen non-specific and generally provides less predictable
levels of disease control. Some BCAs exhibit predatory behavior under
nutrient-limited conditions. However,Trichoderma produce a range of enzymes
that are directed against cell walls of fungi. However, when fresh bark is used
in composts, Trichoderma spp. do not directly attack the plant
pathogen, Rhizoctonia solani. But in decomposing bark, the concentration of
readily available cellulose decreases and this activates the chitinase genes
of Trichoderma spp., which in turn produce chitinase to parasitize R.
solani (Benhamou and Chet 1997).

2. Antibiotic-mediated suppression

Antibiotics are microbial toxins that can, at low concentrations, poison or kill
other microorganisms. Most microbes produce and secrete one or more
compounds with antibiotic activity. In some instances, antibiotics produced by
microorganisms have been shown to be particularly effective at suppressing
plant pathogens and the diseases they cause. Some examples of antibiotics
reported to be involved in plant pathogen suppression are listed in Table 2. In
all cases, the antibiotics have been shown to be particularly effective at
suppressing growth of the target pathogen in vitro and/or in situ. To be
effective, antibiotics must be produced in sufficient quantities near the
pathogen to result in a biocontrol effect. In situ production of antibiotics by
several different biocontrol agents has been measured (Thomashow et al.
2002); however, the effective quantities are difficult to estimate because of

38
the small quantities produced relative to the other, less toxic, organic
compounds present in the phytosphere. And while methods have been
developed to ascertain when and where biocontrol agents may produce
antibiotics (Notz et al. 2001), detecting expression in the infection court is
difficult because of the heterogenous distribution of plant-associated microbes
and the potential sites of infection. In a few cases, the relative importance of
antibiotic production by biocontrol bacteria has been demonstrated, where
one or more genes responsible for biosynthesis of the antibiotics have been
manipulated. For example, mutant strains incapable of producing phenazines
(Thomashow and Weller 1988) or phloroglucinols (Keel et al. 1992, Fenton et
al. 1992) have been shown to be equally capable of colonizing the rhizosphere
but much less capable of suppressing soilborne root diseases than the
corresponding wild-type and complemented mutant strains. Several biocontrol
strains are known to produce multiple antibiotics which can suppress one or
more pathogens. For example, Bacillus cereus strain UW85 is known to produce
both zwittermycin (Silo-Suh et al. 1994) and kanosamine (Milner et al. 1996).
The ability to produce multiple antibiotics probably helps to suppress diverse
microbial competitors, some of which are likely to be plant pathogens. The
ability to produce multiple classes of antibiotics, that differentially inhibit
different pathogens, is likely to enhance biological control. More
recently, Pseudomonas putida WCS358r strains genetically engineered to
produce phenazine and DAPG displayed improved capacities to suppress plant
diseases in field-grown wheat (Glandorf et al. 2001, Bakker et al. 2002).

Some of antibiotics produced by BCAs

Antibiotic Source Target pathogen Disease Reference
2, 4-diacetyl- Pseudomonas Shanahan et
Pythium spp. Damping off
phloroglucinol fluorescens F113 al. (1992)
Agrobacterium Agrobacterium
Agrocin 84 Crown gall Kerr (1980)
radiobacter tumefaciens
Bacillus Aflatoxin Moyne et al.
Bacillomycin D Aspergillus flavus
subtilis AU195 contamination (2001)
Bacillus
Bacillomycin, Fusarium Koumoutsi et
amyloliquefacie Wilt
fengycin oxysporum al. (2004)
ns FZB42
Lysobacter sp. Aphanomyces Islam et al.
Xanthobaccin A Damping off
strain SB-K88 cochlioides (2005)
Trichoderma Wilhite et al.
Gliotoxin Rhizoctonia solani Root rots
virens (2001)
Pantoea
Sandra et al.
Herbicolin agglomerans C9- Erwinia amylovora Fire blight
(2001)
1
Paulitz and
Botrytis Belanger
B.
Iturin A cinerea and R. Damping off (2001),
subtilis QST713
solani Kloepper et
al. (2004)

39
 B. Pythium Leclere et al.
Mycosubtilin Damping off
subtilis BBG100 aphanidermatum (2005)
P. fluorescens 2- Gaeumannomyces Thomashow
Phenazines Take-all
79 and 30-84 graminis var. tritici et al. (1990)
Pythium Howell and
Pyoluteorin, P.
ultimum and R. Damping off Stipanovic
pyrrolnitrin fluorescens Pf-5
solani (1980)
R.
Pyrrolnitrin, Burkholderia Damping off Homma et al.
solani and Pyriculari
pseudane cepacia and rice blast (1989)
a oryzae
Phytophthora
Bacillus Smith et al.
Zwittermicin A medicaginis and P. Damping off
cereus UW85 (1993)
aphanidermatum

3. Lytic enzymes and other byproducts of microbial life

Diverse microorganisms secrete and excrete other metabolites that can

interfere with pathogen growth and/or activities. Many microorganisms
produce and release lytic enzymes that can hydrolyze a wide variety of
polymeric compounds, including chitin, proteins, cellulose, hemicellulose, and
DNA. Expression and secretion of these enzymes by different microbes can
sometimes result in the suppression of plant pathogen activities directly. For
example, control of Sclerotium rolfsii by Serratia marcescens appeared to be
mediated by chitinase expression (Ordentlich et al. 1988). And, a b-1,3-
glucanase contributes significantly to biocontrol activities of Lysobacter
enzymogenes strain C3 (Palumbo et al. 2005). While they may stress and/or
lyse cell walls of living organisms, these enzymes generally act to decompose
plant residues and nonliving organic matter. Currently, it is unclear how much
of the lytic enzyme activity that can be detected in the natural environment
represents specific responses to microbe-microbe interactions. It seems more
likely that such activities are largely indicative of the need to degrade complex
polymers in order to obtain carbon nutrition. Nonetheless, microbes that show
a preference for colonizing and lysing plant pathogens might be classified as
biocontrol agents. Lysobacter and Myxobacteria are known to produce copious
amounts of lytic enzymes, and some isolates have been shown to be effective
at suppressing fungal plant pathogens (Kobayashi and El-Barrad 1996, Bull et
al. 2002). So, the lines between competition, hyperparasitism, and antibiosis
are generally blurred. Furthermore, some products of lytic enzyme activity
may contribute to indirect disease suppression. For example, oligosaccharides
derived from fungal cell walls are known to be potent inducers of plant host
defenses. Interestingly, Lysobacter enzymogenes strain C3 has been shown to
induce plant host resistance to disease (Kilic-Ekici and Yuen 2003), though the
precise activities leading to this induction are not entirely clear. The
quantitative contribution of any and all of the above compounds to disease
suppression is likely to be dependent on the composition and carbon to
nitrogen ratio of the soil organic matter that serves as a food source for

40
microbial populations in the soil and rhizosphere. However, such activities can
be manipulated so as to result in greater disease suppression. For example, in
post-harvest disease control, addition of chitosan can stimulate microbial
degradation of pathogens similar to that of an applied hyperparasite
(Benhamou 2004). Chitosan is a non-toxic and biodegradable polymer of beta-
1,4-glucosamine produced from chitin by alkaline deacylation. Amendment of
plant growth substratum with chitosan suppressed the root rot caused
by Fusarium oxysporum f. sp. radicis-lycopersici in tomato (Lafontaine and
Benhamou 1996). Although the exact mechanism of action of chitosan is not
fully understood, it has been observed that treatment with chitosan increased
resistance to pathogens.

4. Competition

From a microbial perspective, soils and living plant surfaces are frequently
nutrient limited environments. To successfully colonize the phytosphere, a
microbe must effectively compete for the available nutrients. On plant
surfaces, host-supplied nutrients include exudates, leachates, or senesced
tissue. Additionally, nutrients can be obtained from waste products of other
organisms such as insects (e.g. aphid honeydew on leaf surface) and the soil.
While difficult to prove directly, much indirect evidence suggests
that competition between pathogens and non-pathogens for nutrient resources
is important for limiting disease incidence and severity. In general, soilborne
pathogens, such as species of Fusarium and Pythium, that infect through
mycelial contact are more susceptible to competition from other soil- and
plant-associated microbes than those pathogens that germinate directly on
plant surfaces and infect through appressoria and infection pegs. Genetic work
of Anderson et al. (1988) revealed that production of a particular plant
glycoprotein called agglutinin was correlated with potential of P. putida to
colonize the root system. P. putida mutants deficient in this ability exhibited
reduced capacity to colonize the rhizosphere and a corresponding reduction in
Fusarium wilt suppression in cucumber (Tari and Anderson 1988). The most
abundant nonpathogenic plant-associated microbes are generally thought to
protect the plant by rapid colonization and thereby exhausting the limited
available substrates so that none are available for pathogens to grow. For
example, effective catabolism of nutrients in the spermosphere has been
identified as a mechanism contributing to the suppression of Pythium
ultimum by Enterobacter cloacae (van Dijk and Nelson 2000, Kageyama and
Nelson 2003). At the same time, these microbes produce metabolites that
suppress pathogens. These microbes colonize the sites where water and
carbon-containing nutrients are most readily available, such as exit points of
secondary roots, damaged epidermal cells, and nectaries and utilize the root
mucilage.

5. Induction of host resistance

41
Plants actively respond to a variety of environmental stimuli, including gravity,
light, temperature, physical stress, water and nutrient availability. Plants also
respond to a variety of chemical stimuli produced by soil- and plant-associated
microbes. Such stimuli can either induce or condition plant host defenses
through biochemical changes that enhance resistance against subsequent
infection by a variety of pathogens. Induction of host defenses can be local
and/or systemic in nature, depending on the type, source, and amount of
stimuli. Recently, phytopathologists have begun to characterize the
determinants and pathways of induced resistance stimulated by biological
control agents and other non-pathogenic microbes (Table 3). The first of these
pathways, termed systemic acquired resistance (SAR), is mediated by salicylic
acid (SA), a compound which is frequently produced following pathogen
infection and typically leads to the expression of pathogenesis-related (PR)
proteins. These PR proteins include a variety of enzymes some of which may
act directly to lyse invading cells, reinforce cell wall boundaries to resist
infections, or induce localized cell death. A second phenotype, first referred
to as induced systemic resistance (ISR), is mediated by jasmonic acid (JA)
and/or ethylene, which are produced following applications of some
nonpathogenic rhizobacteria. Interestingly, the SA- and JA- dependent defense
pathways can be mutually antagonistic, and some bacterial pathogens take
advantage of this to overcome the SAR. For example, pathogenic strains
of Pseudomonassyringae produce coronatine, which is similar to JA, to
overcome the SA-mediated pathway (He et al. 2004). Because the various host-
resistance pathways can be activated to varying degrees by different microbes
and insect feeding, it is plausible that multiple stimuli are constantly being
received and processed by the plant. Thus, the magnitude and duration of host
defense induction will likely vary over time. Only if induction can be
controlled, i.e. by overwhelming or synergistically interacting with endogenous
signals, will host resistance be increased.

Bacterial determinants and types of host resistance induced by biocontrol agents

Plant Bacterial
Bacterial strain Type Reference
species determinant
Bacillus Peroxidase, chitinase Bargabus et al.
Sugar beet ISR
mycoides strain Bac J and β-1,3-glucanase (2002)
Bacillus subtilis GB03
Arabidopsis 2,3-butanediol ISR Ryu et al. (2004)
and IN937a
Pseudomonas
fluorescens strains
Maurhofer et al.
CHA0 Tobacco Siderophore SAR
(1994)
Arabidopsis Antibiotics (DAPG) ISR Iavicoli et al. (2003)
WCS374 Radish Lipopolysaccharide ISR Leeman et al. (1995)
Siderophore Leeman et al. (1995)
Iron regulated factor Leeman et al. (1995)
WCS417 Carnation Lipopolysaccharide ISR Van Peer and

42
 Schipper (1992)
Radish Lipopolysaccharide ISR Leeman et al. (1995)
Iron regulated factor Leeman et al. (1995)
Van Wees et al.
Arabidopsis Lipopolysaccharide ISR
(1997)
Tomato Lipopolysaccharide ISR Duijff et al. (1997)
Pseudomonas Meziane et al.
Arabidopsis Lipopolysaccharide ISR
putida strains (2005)
Meziane et al.
WCS 358 Arabidopsis Lipopolysaccharide ISR
(2005)
Meziane et al.
Siderophore ISR
(2005)
BTP1 Bean Z,3-hexenal ISR Ongena et al. (2004)
Serratia
Cucumber Siderophore ISR Press et al. (2001)
marcescens 90-166

IV. Evaluation (self-check)

1. Briefly define the type interactions contributing to biological control. (15

points)
2. What are the different mechanisms of biological control? Discuss each by
providing examples. (15 points)

43
Lesson 3
Biological Control Agents of Weeds

I. Overview

Biological control of weeds is the deliberate use of natural enemies to

reduce the density of a particular weed to a tolerable level. The objective of
biological weed control is not eradication but simply the reduction of the weed
population to an economically low level. In fact for biological control to be
continuously successful, small numbers of the weed host must always be
present to assured the survival of the natural enemy.

II. Objectives

At the end of this lesson, the students should be able to:

1. Discuss the methods for biological control of weeds.
2. Determine the different biological agents used for weed control.

III. Lesson Content

A. The Origins of Biological Weed Control

In ancient times, the Chinese discovered that increasing ant populations in

their citrus groves helped decrease destructive populations of large boring
beetles and caterpillars. That use of a natural enemy to control a pest marked
the birth of biological control.

Biological control research and implementation is even more relevant today.

Foreign and native organisms that attack weeds are being evaluated for use as
biological control agents. As a weed management method, biological control
offers an environmentally friendly approach that complements conventional
methods. It helps meet the need for new weed management strategies since
some weeds have become resistant to certain herbicides. Biological control
agents target specific weeds. Moreover, this technology is safe for applicators
and consumers.

By 1925, Australia was struggling with 60 million acres of grazing land heavily
infested with prickly pear cactus. Hundreds of square miles were virtually
impenetrable to humans or animals. A small moth from Argentina was
imported and released. The moth larvae burrowed into the cactus, grew and
multiplied, and within 10 years had decimated the prickly pear population.
Today, the cactus covers only 1% of the area it occupied in 1925.

44
B. The problem with weeds

Weeds can be defined as plants growing out of place. For example,

waterhyacinth is beautiful in floating gardens but can rapidly clog waterways,
making navigation impossible. Similarly, morningglory is beautiful in the
garden, but when it entwines corn stalks, it can destroy a farmer's crop.

1. Weeds degrade native ecosystems.

Invasive, noxious weeds such as leafy spurge and yellow starthistle infest
millions of acres of rangeland and wilderness areas in the northern plains
and are estimated to cost tens of millions of dollars annually in lost grazing
and associated economic effects. For example, leafy spurge and yellow
starthistle spread and form dense stands competing with native plants,
reducing plant diversity, and degrading wildlife habitats.

2. Weeds foul waterways

Purple loosestrife has run rampant in U.S. waterways and natural wetland
ecosystems, choking out cattails and other native aquatic plants that
provide food and shelter for fish, mammals, birds, and reptiles. Many
believe that purple loosestrife seeds arrived in the United States during the
early 1800's in soil used as ship ballast.

3. Weeds lower property values.

Knowledgeable people avoid buying land infested with invasive weeds.
Much time and money will probably be required to convert weed-infested
fields to more productive or aesthetically pleasing land.

C. Biological Control of Weeds

Biological weed control involves using living organisms, such as insects,

nematodes, bacteria, or fungi, to reduce weed populations. In nature, plants
are controlled biologically by naturally occurring organisms. Plants become
pests - and are labeled "weeds" - when they run rampant because their natural
enemies become ineffective or are nonexistent. The natural cycle may be
interrupted when a plant is introduced into a new environment, or when
humans disrupt the ecological system. When we purposefully introduce
biological control agents, we are attempting to restore or enhance nature's
systems.

1. How does it work?

Roots provide plants with water and nutrients. Some biological control
agents attach to roots and thereby stunt plant growth. Some bacteria live
on root surfaces and release toxins that stunt root growth. Many fungi
infect roots and disrupt the water transport system, which reduces leaf

45
growth. Beneficial insects and nematodes feed directly on the weed roots
causing injury which allows bacteria and fungi to penetrate.

Plant leaves capture energy from the sun and store it as sugar. Insects that
feed on leaves reduce the leaf surface available for energy capture. Fungi
and bacteria that infect leaves reduce the ability of the leaf to make
sugars. In either case, there is less energy available for weed growth.
Whether through damage on roots or leaves, severe infestations of
biological control agents can actually kill weeds, reducing their adverse
effects on desirable plants.

Many weed species survive from year to year by producing seeds. Fungi or
insects that attack seeds can reduce the number of weed seeds stored in
the soil, which in turn can reduce the size of future weed populations. This
lowers the effort needed to control the remaining emerging weeds.

Some bacteria and fungi applied as biological control agents do not survive
from year to year. These organisms must be applied on an annual basis.
This technique is called the "bioherbicide" strategy. With this tactic,
biological agents are used a in manner similar to chemical herbicides.

Weeds introduced from foreign countries often require a different strategy.

Insects and pathogens are collected in the area of origin and evaluated for
release in North America. Insect agents often require a number of years to
become fully effective. Their growth is often hindered by adverse climatic
conditions. Long-term monitoring is needed to determine their
effectiveness. The release of biological control organisms in this manner is
termed the "classical" approach to biological control. Fungi that naturally
spread and infect weeds can also be used in a classical biological control
strategy.

2. Biological control is worth the effort

It is well demonstrated that weeds can be controlled biologically.
Deleterious rhizobacteria have been used to stunt weed growth in wheat
fields in the Pacific Northwest. A rust fungus has been used to eliminate
rush skeletonweed from thousands of acres of rangeland in the West. A
complex of introduced insects has also cleared alligatorweed from
waterways, rice fields and lakes in the South.

The cost of developing and conducting a biological control program varies

with the target weed and the strategy selected. On average, a biological
control program will cost about $4 million. But every dollar spent in
development returns at least $50 in benefit.

Biological control of weeds will not eliminate the need to use chemical
herbicides. Both of these tools need to be integrated with cultural

46
 practices, such as tillage and crop rotation, in the battle against weeds. By
using Integrated Weed Management, the development of weeds that are
resistant to biological or chemical agents can be slowed.
D. Methods Used in Biocontrol of Weeds

a. Augmentation of natural enemy populations: Augmentation includes the

periodic release and/or distribution of natural enemies.
b. Application of “biological herbicides”

1. Procedures in Classical Biological Control

a. Determine the suitability of the weed for this approach. Not all weeds
are suitable and those with the following characteristics are generally least
suited for biological control:

i. Weed species which are valued in other situations are not good
candidates for this approach.
ii. Weeds that are closely related to economic crops are not good
candidates for this method. The closer the relationship the less
possibility there is that a biotic agent could distinguish between the
weed and the crop.
iii. Native weed species are not generally amenable to this approach.
iv. Weeds of cropland under intensive cultivation are generally not
suited to this approach.
v. Minor weed problems are not generally suited to this approach.
vi. If eradication of the weed is desired (e.g., poisonous weeds), the
method is generally not applicable.

Therefore, in classical biocontrol of weeds, the ideal target weed is an

aggressive. Introduced weed which infest areas of marginal land such as
rangeland, pastures, and waste areas.

i. Conduct a survey overseas to determine if there are any parasites

available for introduction against the weed.
ii. Determine the potential effectiveness of the parasite in an attempt
to eliminate ineffective agents before importation and screening
tests.
iii. Determine the safety of the selected parasite for release. Very
extensive tests must be conducted in the quarantine facilities.
iv. Establish the biocontrol agent in the area on infestations of the
target weed.

2. Plant Pathogens as Biocontrol of Weeds

Plants pathogens offer two advantages over insects as biocontrol agents of

weeds: 1) they are often more host specific, and 2) they can be applied with

47
conventional spray equipment at a time when the weed is at its most
susceptible stage.

a. Conventional approach
Conventional methods of weed control involve the use of specially
formulated chemical compounds to interrupt specific biochemical pathways
(called modes of action, or MOA) in order to eliminate weeds without
affecting crops being grown. The herbicides used depend on the type of
weed being targeted, the developmental stage of these weeds, and the
crops being raised.

b. Biological Herbicides
Bioherbicides are phytopathogenic microorganisms or microbial phytotoxins
useful for biological weed control applied in similar ways to conventional
herbicides. The active ingredient in a bioherbicide is, however, a living
microorganism. Most commonly the organism is a fungus; hence the term
mycoherbicide is often used in these cases. Although the use of fungi and
bacteria as inundative biological control agents (bioherbicides) has been
recognized as a significant technological weed control alternative, it can
be argued that it serves a more important role as a complementary
component in successful integrated management strategies, and not as a
replacement for chemical herbicides and other weed management tactics.
Actually, in many situations, bioherbicides can be used as the sole option
for the management of one or two target weeds, i.e. as a minor
supplement to conventional chemical herbicides.

IV. Evaluation (self-check)

1. Why weeds became a problem? Give at least 5 reasons and discuss each.
(15 points)
2. Enumerate 10 examples of bioherbicides and list the weeds it controls. (50
points)

48
49