Sorsogon State College

Castilla Campus
Beneficial Arthropods and Microorganism
Module 6

Topics: Nature of Biological Control Agent (Pathogen)

Week:

Target Learner: BSA AgEd 3

Estimated Time Duration

I. Introduction

In previous module, you learned the nature of predators as biological agent. In

this module, you will be learning the nature of pathogen as biological agent.
A variety of microbial pathogens including bacteria, protozoans, microsporidia,
viruses and fungi are specifically pathogenic to insects and completely harmless to
other forms of life. This selective pathogenicity renders many of them valuable as
biological control agents of insect pests. Some, such as Baccilus thuringiensis, have
been the source of natural insect toxins that are now synthesized as biopesticides,
or engineered directly into crop plants.

II. Module Objectives

By the end of this module, you are expected to do the following:

1. Define pathogen
2. cite example of beneficial microbial pathogen
3. Describe the signs and symptoms of insect infection
4. Describe the characteristics of insects infected with microbial pathogen

III. Learning Resources

Microbial Pathogen

Pathogen are microorganisms that cause disease in insects. These include

bacteria. Viruses, fungi,nematodes and protozoa. They are an important part of natural
control. Spontaneou epizootics of pathogens sometimes occur in pest populations (
Fuxa and Tanada, 1987 as cited by Van Driesche et al, 2008) like for example the viral
and fungal epidemics that periodically decimate gypsy moth larvae( Lymatria dispar L.)
in North America ( Gillock and Hain 2001, 2002 as cited by Van Driesche et al, 2008)
The use of pathogens in biological control has included programs against the rhinoceros
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beetle( Oryctes rhinoceros L.) on coconut by introduction of an Oryctes virus or the use
of Bacillus Thuringiensis Berliner for the control of lepidopterous pest in corn.

1. Bacteria

Bacteria have been the most successfully brought into commercial use. Most
emphasis has been placed on bacillus thuringiensis Berliner which has at least 65 known
subspecies.

Generally, bacteria that do not form endospores( more commonly called spores)
such as Pseudomonas, Aerobacter, Cloaca or Serratia usually cause disease only when
the host is stressed. On the other hand , endospore forming bacteria can more easily
infect healthy host through ingestion of the endospores. Species like B. thuringiensis B.
sphaericus Meyer and Neide and Paenibacillus popilliae Dutky ( formerly in Bacillus)
Petterson et al, 1999 as cited by Driesche et al, 20008) are the pathogens that have
most often been used for biological control

Infections by [Link] Dutky on the Japanese beetle, popillia japonica Newman

are referred to as milky spore disease because the hemolymph of the diseased host
usually exhibits a milky color B. sphaericus Meyer and Neide, which kills mosquito
larvae, has its insecticidal activity due to crystalline toxins that are released when the
insect digest spores it has taken in with its food B. Thuringiensis, the most extensively
marketed bacterial pathogen of insects, have 65 serotypes recognized and given
subspecies names. Most of the serotypes affect caterpillars and some have been used
against various lepidopteran pests of fruits and vegetable. Other subspecies, israelensis
and tenebrionis are effective against fly larvae and chrysomelid beetles respectively.
Insects that ingest the endospores are killed due to the combined effects of poisoning
by toxins and bacterial multiplication

2. Fungi

Over 400 species of fungi that infect insects have been recognized ( hall and
Papierok, 1982 as cited by Van Driesche et al, 2008) Most attention has focused on
about 20 species in 12 genera( Roberts and Wraight, 1986 as cited by Van Driesche et
al, 2008) These include Lagenidium, Entomophaga, Neozygites, Entomophthora, Erynia
, Aschersonia, Verticilium, Nomuraea, Hirsutella, Metarhizium, Beauveria and
Paecilomyces. Lagenidium infects mosquito larvae and does not need an alternate host
to complete its life cycle. The fungi belonging to Entomophthoraceae. Composed of
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Entomophaga, Entomophthora, Neozygetes and Erynia are important as natuarally
occurng pathogens. Host of this groupinclude caterpillars, beetles aphids and mites
Aschersonia, Verticillium, Nomuraea, Hirsutella, Metarhizium, Beauveria and
Paecilomyces are all Imperfect Fungi(Those without sexualstage/forms). Hersutella
thompsonii Fischer is a pathogen of eriophyid rust mites; Beauveria bassiana (
Balsalmo) Vuillemin has a wide host range; Metarhizium and Aschersoniaspecies have
been studied as pathogens of whiteflies aphids and scale( Van Driesche et al, 2008)

3. Viruses

Baculoviruses usually kill their host and are only known to infect insects this
family contains the nucleopolyhedroviruses ( NPV) and granuloviruses(GV)
Baculoviruses may act as natural pathogens, causing periodic cycle of disease in
insects. The level of control is however rarely high unless virus levels are augmented
artificially.

Baculoviruses may also be formulated as biopestices. Since viruses are obligate

parasites they are reaed in living insects or in insect cell culture.

Signs and symptoms of insect infection

A Movement and irritability. This can be demonstrated by inoculating live insects

and then observed for movement and response. The diseased insect shows abnormal
and uncoordinated movements. Infected insects may also show lack of response or
abnormal responses to stimuli

B. Discoloration. This is also called melanization. When there is bacterial infection

there is a change from brown to dark coloration. There is also breakdown of tissues.
Sometimes bactreia pass pigments which are important to insects for example, serratia
marcescens Bizio

.C. Changes in size and shape. This can be shown in either by the following:

Stunted or smaller size for diseased than healthy insects

-BT can cause gut paralysis so insects stop feeding
Inflated- insect become bloated due to microorganisms
- Fungi forming sporangia cause swelling inside; some organs are also enlarge
Tumor like appearances or protrusions
Abnormal change in consistency( brittle, soft, cheesy and hard)
Presence of either macroscopically
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D. Disturbance in the digestive system. This can be observe due to reduce
appetite of the insects; vomiting due to regurgitation of its food and watery fluid is
coming out of the mouthparts. THRE IS ALSO DIARRHEAL SYSTOMS OF THE INFECTED
INSECTS

E.) Other abnormal reaction in physiological process. Some of these may be

demonstrated by non pupation and non emergence. In extreme cases infected insect
failed to develop and do not molt. There are reproductive disturbances of the infected
individuals such as non laying of female or failure to oviposit fertile eggs, reduced
fecundity or even sterility .

Characteristics of insects infected with the following:

1. Viruses

Infected insects will go up on the top of the vegetation and die hanging with prolegs.
This is very relevant for dispersal of the pathogen since it facilitates wider dissemination
or showering of spores downward. The body content will appear porcelain white; the
body has bluish purple iridescence(shine) and there is rapid liquefaction of the body
contents

Insects infected with nuclear polyhedrosis viruses( NPV) will exhibit several symptoms
and changes. The larval skin darkens and may have yellow patches or appears oily; the
skin becomes fragile and the hemolymph becomes turbid. Prior to death, the infected
larva usually climbs to highest point of the vegetation available and dies. After death,
the intergument frequently raptures, releasing millions of polyhedral viruses.

2. Bacteria-soft or dried /scale

Infected insects have cheesy or with sticky consistency. Intergument darkens with an
accompanying odor. Some infected insects turn milky white in color or turn red and
sometimes hanging on the top of vegetation with their prolegs
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3. Fungi

Infected insect body becomes mummified and may have cheese like body contents. The
body is filled or covered with filamentous hyphae. Some fungi can be distinguished by
aerial growth; for example, cordyceps have horny conidial outgrowth. Body may
become black, hollow and disintegrating into a black powder or could turn re, like
serratia that has 3 genera causing red [Link] aquatic insects, infection is evident
because infected insects have spherical cyst like sporangia. For example, infection of
Coelomomyces in mosquitos is shown by the anal papillae become filled up with
sporangia other example are grasshoppers found embracing on the stem of host which
shows fungal infection. A cicada may have its abdomen disintegrating due to infection
with genus Massospora but stil capable of locomotion. Some flies become stuck on the
window ane with springler of white structure like halo when infected with the genus
Entomopthora. More than 36 different genera of fungi contain species which cause
insect diseases however, identification of fungus is difficult. Most fungi are transmitted
from one host to another by spore usually a conidium

The characteristics exhibited by fungi infected insects are cessation of feeding

weakness and disorientation; change in color and the cuticle may show dark spots due
to areas of fungal penetration and presence of mycelia in or on the affected area

How do fungi kill the host?

Asphyxiation or starvation( e.g, Zygomycotina)

Toxins released by the fungus

Uptake of host nutrients and water results in desiccation (Mummification)of the insects
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Examples of Entomopathogenic fungi

a. Zygomycotina:

Oerder Entomopathogenic Fungi

Batkoa

Entomophthora

Entomophaga

Erynia

Neozygites

Zoophthora

b. Deuteromycotina:

Order Moniliales

Beauveria bassiana Vuillemin-whie muscardine, produce natural toxins called

beauvericin

Metarhizium anisopliae(Metschnikoff) Sorokin- green muscardine; produces the toxin

destruxins

Lecanicillium(verticillium) lecanii( Zimmerman) Zare et W. Gams- infects mostly aphids

and scale insects

cabbage fly killed by beauveria bassiana

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Metarhizium anisopliae infected grubs of Oryctes rhinocerus (Photo: Central Plantation

Crops Research Institute [CPCRI] India

Protozoans

The infected insects are with morphological deformities or remain small in size
due to reduced feeding. They are lethargic and may have difficulty in molting and lose
balance. They produce white fecal exudates. The aquatic hosts show a transparent
cuticle often with white opaque or swollen areas. The infected terrestrial insects exhibit
black spots on their integument eventually, the infected insects collapse and die.

An example is Nosema which infects grasshoppers. The spores are microsporida that
possess polar filament which injects infective material from the spore into the host cell.
It is a long tube coiled within the spore and extruded once inside the body of the host.
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a Hirsutella citriformis growing on a mummified adult Diaphorina citri. b The same H.

citriformis isolate growing on mycological media after approximately five weeks

Nematodes

Class Adenophorea: Stichosomida

Mostly free living aquatic form few insect parasites

Large group of obligate parasites
Ramanomermis culicivorax- pathogen of early instar mosquito larvae which are
killed before pupation

Mode of infection:

Eggs laid at bottom of mosquito pool to hatch

Preparasitic larva penetrates an early instar mosquito

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Nematodes often migrate to the thorax, quickly obtaining nourishment from the insect
hemolymph

Emerges from the last instar mosquito larva, killing it before pupation

mosquito wriggler infected with mermithid nematode

Class Secernantia: Rhabditida

Family Steinernematidae

Contain groups which are entomogenous. Example are Steinernema(= Neoaplectana)

and Heterorhabditis

Mode of infection

The free living 3rd stage penetrates the susceptible host. They are attracted chemically
through the mouth or anus and enter the hemocoel via the gut wall. The larvae carry
highly virulent bacteria( Xenorhabdus spp.) which are released into the haemocoel and
cause septicemia nematodes feed decomposing tissues containing the bacteria and then
the insect dies. Nematode is visible through the transparent integument of the infected
insects.

IV. Assessment
Activity 1.
Direction: Read and study the following questions below. You may use a separate
document to answer this activity

1. Why it is important microbial pathogen as biological agent? Give 5 example of

beneficial microbial pathogen?
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Beneficial Arthropods and Microorganism
2. Describe the characteristics of insect infected with specific bacteria, fungi and
virus. Give 3 example each microbial pathogen

Activity 2

Write your answer on the blank

1. ______________ Virus biological control against the rhinoceros beetle(

Oryctes rhinoceros L.) on coconut
2. _______________ Bacteria biological for the control of lepidopterous pest in
corn.
3. ________________ referred to as milky spore disease
4. ________________ Bacteria that effective against fly larvae and chrysomelid
beetles
5. ________________ usually kill their host and are only known to infect insects
.This family contains the nucleopolyhedroviruses and granuloviruses.
6. ________________ The larval skin darkens and may have yellow patches or
appears oily; the skin becomes fragile and the hemolymph becomes turbid
are characteristics infected with?
7. _______________ Infected insect body becomes mummified and may have
cheese like body contents
8. _______________ produce natural toxins called beauvericin
9. _______________ produces the toxin destruxins
10. ________________ The infected insects are with morphological deformities
or remain small in size due to reduced feeding

V. References

[Link]
adult-Diaphorina-citri-note-the_fig1_257523929

[Link]
bassiana-sl-a-Nymph-of-the-silverleaf_fig1_308172345

Beneficial Arthropods lecture and Microorganism manual, Visayas State University