Biological Control 1
Biological Control
Leslie C. Lewis, Agricultural Research Service, USDA, Ankeny, Iowa 50021, and Iowa State University,
Ames, Iowa 50011, United States
1. Introduction . . . . . . . . . . . . . . .
1.1. Entomophagous Insects . . . . . . . .
2. Biological Control of Insects . . . . .
2.0.1. Parasitoids . . . . . . . . . . . . . . . . .
2.0.2. Insect Predators . . . . . . . . . . . . . .
2.1. Other Predators . . . . . . . . . . . . .
2.2. Entomopathogens . . . . . . . . . . . .
2.2.1. Bacteria . . . . . . . . . . . . . . . . . . .
2.2.2. Pathogens with Protozoal Characteris-
tics . . . . . . . . . . . . . . . . . . . . . .
2.2.3. Insect Viruses . . . . . . . . . . . . . . .
2.2.4. Fungi . . . . . . . . . . . . . . . . . . . .
2.3. Nematodes . . . . . . . . . . . . . . . .
3. Biological Control of Weeds . . . . .
3.1. Phytophagous Insects . . . . . . . . .
3.2. Plant Pathogens . . . . . . . . . . . . .
1. Introduction
Biological control is the action of natural ene-
mies in maintaining another organism’s popu-
lation density at a lower level than would oth-
erwise occur. In this article, however, the tra-
ditional definition has been expanded to in-
clude chemicals that duplicate or mimic natu-
ral ones. The major effort in biological control
has been focused on the suppression of popula-
tions of pest insects. However, biological con-
trol of weeds and plant pathogens is a very ac-
tive area of research. Biological control tech-
niques also have been used successfully to sup-
press populations of animals other than insects.
Biological control is a global endeavor; special-
ists in this discipline have developed a world-
wide organization, International Organization
for Biological Control of Noxious Animals and
Plants (IOBC). More information on IOBC may
be obtained by contacting: Secretary-General,
G. Mathys, 1, rue Le Nôtre, F-75016 Paris.
Classical biological control involves: (1) dis-
covery, (2) importation, (3) release, and (4) es-
tablishment of exotic species of natural ene-
mies. Most pest insects, especially in the United

c 2005 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim
10.1002/14356007.a04 077
1 3.3. Higher Animals . . . . . . . . . . . . . 12
2 4. Biological Control of Plant
3 Pathogens . . . . . . . . . . . . . . . . . 12
3 4.1. Macroorganisms . . . . . . . . . . . . . 12
4 4.2. Microorganisms . . . . . . . . . . . . . 12
5 5. Other Biological Methods of Control 12
5 5.1. Biochemicals . . . . . . . . . . . . . . . 12
5 5.1.1. Growth Regulators . . . . . . . . . . . . 12
5.1.2. Semiochemicals . . . . . . . . . . . . . . 14
7 5.1.3. Bioderived Toxicants . . . . . . . . . . 15
8 5.2. Autocidal . . . . . . . . . . . . . . . . . 15
9 5.2.1. Sterile Male . . . . . . . . . . . . . . . . 15
10 5.2.2. Hybrid Sterility . . . . . . . . . . . . . . 16
11 6. Formulation, Application,
11 and Efficacy . . . . . . . . . . . . . . . . 16
11 7. References . . . . . . . . . . . . . . . . . 19
States, are pests because they were introduced
accidentally, or on purpose, leaving their com-
plex of natural enemies behind. For example,
several thousand species of insects have been
introduced into the United States in attempts to
control a few species of insect pests.
Applied biological control is sometimes im-
plemented when a natural enemy does not be-
come established. This involves: (1) augmenta-
tion and (2) conservation. Augmentation is the
manipulation of a natural enemy, and conserva-
tion is the manipulation of the environment, both
to enhance the impact of the natural enemy on
the pest population. Examples of augmentation
are colonization and periodic release of natural
enemies and improvement of the efficiency of a
natural enemy through selection. Conservation
involves, e.g., using different cultural practices
that serve to protect the natural enemy in the
environment.
Natural enemies are: (1) parasitoids, (2)
predators, and (3) pathogens. A parasitoid (par-
asite) is an insect that lays an egg on, or in, a
single host insect. The parasitoid develops inter-
nally, killing the host. Predators are free-living
organisms that kill and consume their prey im-
2 Biological Control
mediately or within a short period of time. Preda-
tory insects kill and consume more than one
host or prey during their development. Preda-
tors are found in all major orders of insects. Par-
asitoids and predators are referred to as ento-
mophagous insects. Pathogens are microorgan-
isms that cause diseases in insects. Most insect
diseases are caused by bacteria, fungi, proto-
zoa, microspora, and viruses. These organisms
are collectively referred to as entomopathogens.
The phylum Nematoda consists of a group of
round worms called nematodes. Some nema-
todes are natural enemies of insects; some ne-
matodes behave like parasitoids, whereas others
behave like insect pathogens. These nematodes
are referred to as entomophagous nematodes.
In the biological control of weeds, natural
enemies usually are phytophagous insects and
microorganisms that cause diseases of plants.
Natural enemies of plant pathogens include
predacious insects that feed on the fruiting bod-
ies of pathogenic fungi and microorganisms that
are parasitic on other microorganisms.
History. Several references are made to early
examples of natural enemies being used to con-
trol pests. For example, in the year 1200, Chinese
citrus growers purchased nests of the predacious
ant, Oecophylla smaragdina, and placed them in
citrus trees to control a caterpillar, Tessarotoma
papillosa. Many other examples of successful
biological control are recorded, including the
introduction to California of the vedalia beetle,
Rodolia cardinalis, in 1888 to control the cot-
tony cushion scale, Icerya purchasi [24]. This
project saved the citrus industry of California
and was the impetus for additional biological
control projects against insect pests.
Widespread adoption of host-specific biolog-
ical control techniques was slowed by the de-
velopment of relatively inexpensive synthetic
chemical insecticides that had a broad spectrum
of insect targets. These compounds were envi-
sioned to be the permanent solution for problems
with insect pests.
The public concern for environmental con-
servation that has surfaced over the past 25
years has again stimulated interest in, and fi-
nancial support for, biological control programs.
During this time, intensive research has been
conducted in the area of insect biochemistry;
specifically, insect hormones and semiochem-
icals. Hormones are chemicals that are pro-
duced in certain body tissues, are transported
within the body by the circulatory system, and
elicit a response in another part of the body
(→ Hormones). Semiochemicals also are pro-
duced within the body but are emitted externally
to elicit a response. The hormones of interest in
biological control are those that regulate growth
and molting of the insect; the semiochemicals
are those that elicit a sexual response or attract
an insect to a point source. The use of biochem-
icals, such as hormones and semiochemicals, is
not a method of biological control in the strict
sense. These chemicals, plus male-sterile tech-
niques, host plant resistance, and genetic ma-
nipulation, are merely other biological methods
used to control insects (see Chap. 5).
Entomologists have determined the eco-
nomic threshold or economic injury levels for
many important insect pests on many agricul-
tural crops. These data allow a grower to de-
termine how many insects can be tolerated by
a crop before control is economically feasible.
In developing this information, researchers have
become aware that complete control or elimina-
tion of a pest is not necessary and that measures
less drastic than a chemical insecticide, such as
biological control, can be effective in protecting
crops. This has brought about the present-day
approach to pest control known as integrated
pest management where a holistic approach is
taken in suppressing pest populations. Briefly,
this involves the assessment of a pest problem
and the judicious use of two or more methods
of control (parasitoids, chemical insecticides,
predators, pathogens, hormones, semiochemi-
cals, plant resistance, and cultural practices) in
a compatible system that will reduce the pest
population to an acceptable level. At all times,
attention must be paid to using sound ecologi-
cal principles so as not to disturb the beneficial
regulatory factors present in nature.
1.1. Entomophagous Insects
An ecosystem has a relationship between a host
(plant, animal, or commodity), insects feeding
on the host, and organisms feeding on the in-
sects. An example of this relationship is illus-
trated in Figure 1.

Figure 1. Example of an ecosystem
2. Biological Control of Insects
2.0.1. Parasitoids
The great majority of parasitoids are found in the
insect orders of Strepsiptera, Coleoptera, Lepi-
doptera, Hymenoptera, and Diptera, with by far
the most being classified as Hymenoptera and
Diptera. To be successful, a parasitoid should
have the following attributes: (1) good searching
ability, (2) high biotic potential, and (3) ability
to operate effectively throughout the entire geo-
graphic range of its host. A typical female par-
asitoid selects a suitable host, deposits a single
egg either on or within it, and leaves in search of
other suitable hosts. The parasitoid egg hatches
and develops within the host insect. During the
early stages of development, the parasitoid does
not greatly disturb the host insect. Later, as the
parasitoid reaches maturity, the host insect be-
comes immobile, at which time the parasitoid
will leave the host’s body and pupate. Shortly
afterward, the host usually dies.
There are many variations in the parasitoids
as to developmental stage of host most desir-
able for parasitization. Some parasitoids prefer
young larvae, some older larvae, and some even
the pupal stage. Other parasitoids parasitize only
insect eggs.
An example of a “generalized” hy-
menopteran parasitoid is Bathyplectes curculio-
nis, imported from Italy to assist in controlling
the alfalfa weevil, Hypera postica. Some par-
asitoids deposit several eggs in a host insect.
Biological Control 3
These eggs hatch, and then at maturity several
hymenopterous pupae are seen attached to the
surface of the host insect. Trichogramma spp.
are well-known as egg parasitoids. The females
of these minute insects deposit one or more eggs
within the eggs of different species of insects
(Fig. 2). There is sufficient nutritive material
present within the egg to support the develop-
ment of this small parasitoid.
Figure 2. Trichogramma wasp preparing to oviposit in an
insect egg
∗ Courtesy of Gerald R. Carner, Clemson University
Some Hymenoptera possess a biological
modification known as polyembryony, the de-
velopment of several individuals from a sin-
gle egg. In this situation, an adult female, e.g.,
Macrocentrus grandii (Fig. 3), a parasitoid of
the European corn borer, Ostrinia nubilalis, de-
posits a single egg in a second-stage corn borer
larva. The primary embryonic germ divides by
fission to form two secondary germs which split
into a variable number of morulae, each of which
develops into an embryo.
Dipterous parasitoids exhibiting a general de-
velopmental cycle are represented by Winthemia
quadripustulata, a parasitoid of the armyworm,
Psuedaletia punctata. The adult female deposits
an egg directly behind the head capsule of a suit-
able host. The parasitoid larva hatches from the
egg and chews its way into the body tissue of
the host, develops to a mature larva, chews its
way out of the larva, and pupates. A dipterous
parasitoid with a different biology is Blepharipa
4 Biological Control
prantensis, a parasitoid of the gypsy moth, Ly-
mantria dispar. The female parasitoid deposits
its egg on the foliage where gypsy moth lar-
vae will feed. The parasitoid is fully developed
within the egg, and the slight pressure exerted
by the mandibles during feeding causes the par-
asitoid to break out of the egg shell.
Figure 3. Adult female Macrocentrus grandii, parasitoid of
the European corn borer, Ostrinia nubilalis
Bonnetia comta (Fig. 4), a parasitoid of sev-
eral cutworms, deposits eggs on host fecal ma-
terial. These eggs immediately hatch, and the
neonates or planidia rest on their freshly cast
egg chorion waiting for their host to move.
Figure 4. Adult female, Bonnetia comta, parasitoid of sev-
eral cutworm larvae
Still another unique modification is the
larvipositing phenomenon exhibited by Lydella
thompsoni, a parasitoid of O. nubilalis. The adult
female searches for entrance holes that tunneling
larvae of the European corn borer have made and
deposits live larvae in the fecal material voided
by the feeding corn borer larva. The parasitoid
larvae actively search for and enter the host by
chewing through the insect cuticle.
These few examples of parasitoids give only a
brief overview of the fascinating and sometimes
complex biology of insect parasitoids.
2.0.2. Insect Predators
Predacious insects differ from parasitoids in that
a predator consumes several insect forms during
its life cycle. Also, in many instances, both the
immature and adult stages of the insect are ac-
tive predators. Most orders of insects contain at
least one family of predators.
The scope of this article does not allow a treat-
ment of examples from all orders of insects; only
a few predators that a home gardener or gen-
eral naturalist would most likely recognize are
described. The order Coleoptera (beetles) con-
tains by far the greatest number of predacious
species. The ubiquitous lady beetle (Fig. 5) has
been observed by many. The female lady bee-
tle lays its eggs in clusters in a habitat near a
food source. The newly hatched larva is a vo-
racious feeder, consuming insect eggs, aphids,
and small immature insects. The adult lady bee-
tle also is predacious, devouring a general vari-
ety of prey. However, some of these coccinelid
beetles are very specific in their prey, i.e., the
vedalia beetle, Rodolia cardinalis, feeds only on
the cottony cushion scale, Icerya purchasi. The
ground beetles are another group of ubiquitous
Coleoptera that are general predators, feeding
on insect eggs, caterpillars, and even the adult
moths of some species. There also are many
predacious aquatic beetles that prey on almost
any aquatic form of life that can be captured.
Figure 5. Adult lady beetle, Coleomegilla maculata
Dragonflies and damselflies (order Odonata)
frequent country ponds in the summer. The im-
mature forms of these insects are aquatic and are
predators of pond life. The adults capture their
prey in flight. Depending on the size of the preda-
tors, the prey ranges from mosquitoes to adults
of the orders Lepidoptera and Hymenoptera.

Another common predator found near water
is the robber fly (order Diptera). The larvae feed
on pest insects in the soil, whereas the adults
capture flying insects in flight. Their prey con-
sists mainly of mosquitoes and midges that are
readily found in an aquatic environment. Syr-
phid larvae, also in the order Diptera, feed on
aphids.
Most members of the order Neuroptera
are predators. A representative is the green
lacewing. The larvae of this insect feed on
aphids, mealybugs, thrips, and other small in-
sects. These larvae also are known to prey on
the eggs of several lepidopterans. The adult
lacewings prey on aphids and mealybugs if they
feed on insects at all.
The praying mantid is predacious in both the
immature and adult stages. These insects prey
on almost any insect that they can overcome.
They frequent flowers and capture insects visit-
ing the flowers for nectar. The Chinese mantid,
Tenodera aridifolia sinensis, is frequently made
available commercially for the home gardener.
2.1. Other Predators
Insects are a main ingredient in diets of several
birds, rodents, fish, amphibians, reptiles, and
arachnids. For example, several birds, including
the downy woodpecker, crow, robin, red-winged
blackbird, purple grackle, chickadee, starling,
and ring-necked pheasant, are known predators
of the European corn borer [25].
2.2. Entomopathogens
Insect pathogens (entomopathogens) are mi-
croorganisms that cause diseases in insects.
These pathogens can cause diseases in benefi-
cial insects as well as pest insects. In this article,
only diseases of pest insects will be addressed.
2.2.1. Bacteria
Entomopathogenic bacteria can be divided into
three groups – those that fit the description of
classical biological control (see Chap. 2), those
that fit the description of applied biological con-
trol, and those that fit into either group. Regard-
less of the category in which these bacteria are
Biological Control 5
classified, they have one thing in common, they
can be grown in an insect and/or in a bacterio-
logical medium. The most prominent bacteria in
biological control are in the genus Bacillus, the
group of gram-positive, mobile, spore-forming
rods.
Bacillus thuringiensis is the most widely
known and researched bacterium within this
group and is differentiated from other spore-
forming bacilli by the presence of a paraspo-
ral body that is formed within the sporangium
during sporogenesis. The parasporal body is a
high-molecular-mass protein crystal that is re-
ferred to as crystalline protein, δ-endotoxin, as
well as a parasporal body. This protein moi-
ety possesses some of the insecticidal properties
of the bacterium [26]. Certain subspecies of B.
thuringiensis under specific growing conditions
produce several other toxins with insecticidal ac-
tivity: α-exotoxin, β-exotoxin, and γ-exotoxin.
The δ-endotoxin, however, is the most important
toxin relative to insecticidal activity and thus is
the one most studied.
The δ-endotoxin is produced commercially
in submerged culture under the conditions de-
scribed in the patent [27]. Fermentation medium
is crucial to successful production. It primarily
contains an energy source (carbohydrate) and a
nitrogen source. Common sources of carbohy-
drate are hydrolyzed corn products, starch, dex-
trose, and molasses. Sources of nitrogen are fish-
meals, cottonseed flour, corn steep liquor, soy-
beans, autolyzed yeast, and casein. Trace min-
erals, namely Mg2+ , Mn2+ , Fe2+ , Zn2+ , and
Ca2+ , are usually added to the medium.
Bacillus thuringiensis is primarily a pathogen
of lepidopterans. A general infection cycle is
illustrated in Figure 6. To elicit its effect, the
crystalline protein must be consumed by a sus-
ceptible insect larva. The protein crystal is actu-
ally a protoxin that is hydrolyzed by enzymes in
the gut of susceptible insects, releasing the pure
toxin. The toxin causes paralysis of the gut. The
insect either starves to death, or the midgut ep-
ithelial cells are damaged, allowing the gut con-
tents, including B. thuringiensis spores, to enter
the hemocoel. At this time, a general infection
occurs.
Most Lepidoptera are susceptible to the δ-
endotoxin, but there are basic differences in the
6 Biological Control
Figure 6. Infection cycle of the bacterium Bacillus thuringiensis
pathological responses of insects to this toxin.
Researchers have divided lepidopterous larvae
into four groups according to response [28]. The
first group, designed Type I, shows a rapid gen-
eral gut paralysis resulting in death 1 – 7 h after
ingestion of the crystal toxin. Type II larvae are
characterized by midgut paralysis a few min-
utes after ingestion of crystals and by cessation
of feeding, but not by general paralysis. Type III
larvae have to ingest both spores and crystals,
do not exhibit either general or gut paralysis,
but do stop feeding. Affected larvae enter a mor-
bid state of varying duration with a fairly slow
bacterial multiplication and finally, with a lethal
infection. Type IV larvae are not susceptible to
the crystal toxin or spores but are susceptible to
the thermostable exotoxin(s).
Within these categories, the definitive re-
sponse is dependent on the age of the insect. For
example, the European corn borer is a Type III
larva responding to a combination of spores and
crystals. However, pure crystals are highly toxic
to neonate larvae and moderately toxic to 6-d-
old larvae. A combination of spores and crystals
is necessary to cause maximum larval mortality
[29].
Not only are the insects categorized relative to
their response, but also isolates are divided into
subspecies based on the antigenic properties of
the flagella that are present during the vegetative
stage of growth [30], [31]. Some subspecies are
further divided by the serology of the crystalline
protein [32]. With use of one or both of these
techniques, > 20 subspecies of B. thuringiensis
have been identified.
Insects susceptible to B. thuringiensis do not
necessarily respond the same to all subspecies
of B. thuringiensis. Likewise, a subspecies of B.
thuringiensis does not have the same virulency
to all susceptible insects. A bioassay technique
is used to quantify the toxicity or potency of the
many isolates of the several subspecies.
Bacillus thuringiensis subspecies israelensis
was isolated from a mosquito breeding site in Is-
rael [33]. The discovery of this subspecies pro-
vided researchers with an isolate with consis-
tently high toxicity against larvae of mosquitoes
and blackflies (order Diptera). All other sub-
species are predominantly toxic for larvae of
Lepidoptera. This subspecies also contains a
parasporal crystal protein. However, this crystal
is irregular in size and instead of a single crystal,
there are 3 – 5 crystals. The pathology caused
by subspecies israelensis is similar to that of
other subspecies of B. thuringiensis in suscepti-
ble Lepidoptera. The pathology in black flies is
most likely very similar. Bacillus thuringiensis
subspecies israelensis is also produced by sub-
merged culture.
β-Exotoxin [23526-02-5], the other toxin
produced by B. thuringiensis, is being empha-
sized as an insecticide.
β-Exotoxin

β-Exotoxin is a nucleotide produced during
the vegetative stage of bacterial growth [34]. It
is composed of adenine, ribose, glucose, and al-
laric acid with a phosphate group. β-Exotoxin
is heat stable and is produced by subspecies
thuringiensis, kenyae, morrisoni, tolworthi, and
darmstadiensis [35]. These subspecies produce
different quantities of exotoxin, depending on
the culture medium. Also, indications are that
some subspecies produce more than one exo-
toxin. Commercial production of β-exotoxin is
carried out under patent US 3758383 [36].
Bacillus popilliae causes a “milky disease”
in some beetles in the family Scarabaeidae,
but most noted in the Japanese beetle, Popil-
lia japonica. Bacillus popilliae is eaten by the
grub; the spore germinates and penetrates the
gut wall. Once in the hemocoel, the bacterium
readily multiplies (septicemia), giving the grub
a whitish appearance visible through the cuticle,
thus the name milky disease.
Bacillus popilliae readily develops in larvae
of the beetle, but microbiological techniques
have not been developed to induce B. popilliae to
sporulate in vitro; thus, all production is carried
out by inoculating Japanese beetle larvae and
harvesting the infected larvae. This technique
has a drawback because the Japanese beetle is
not easily grown in the laboratory.
Bacillus sphaericus is a spore-forming rod
that is readily found in nature. It has been iso-
lated from mosquito larvae, and much research
has been conducted on several strains of this bac-
terium that are insecticidal to several species of
mosquitoes. Bacillus sphaericus does not pro-
duce a parasporal body or β-exotoxin as does
B. thuringiensis. However, there is a proteina-
ceous toxin located in the spore coat. Susceptible
mosquito larvae consume the spore and partly di-
gest the spore wall, releasing a toxin. This toxin
causes feeding inhibition, swelling of the midgut
cells, body tremors, and then death. Death can
occur as soon as 0.5 h after consumption of the
spores. Like B. thuringiensis, B. sphaericus is
produced in submerged culture.
Biological Control 7
2.2.2. Pathogens with Protozoal
Characteristics
Microspora. The phylum Protozoa has been
split into seven distinct phyla [37]. This phy-
lum contains the order Microsporida, intercel-
lular parasites found essentially in all orders of
insects, with new species being described and
host records being published frequently. Patho-
logically, microsporidia differ greatly from other
disease-causing organisms in that they generally
do not cause immediate death of the host. A mi-
crosporidian infection usually is chronic. There
are no specific symptoms to describe insects in-
fected with a microsporidium. The infected in-
sect usually is lethargic, has reduced feeding, is
small, has morphological deformities, and fails
to molt or pupate.
Although microsporidia are widely dis-
tributed in the insect world, most microsporidia
infect a single, or at most, a very few hosts.
Nosema pyrausta is a microsporidium of this
type and is a classical biological control agent.
Nosema pyrausta predominantly infects larvae
of the European corn borer, Ostrinia nubilalis.
A schematic illustration of the relationship bet-
ween N. pyrausta and O. nubilalis is presented
in Figure 7. Nosema pyrausta has a life cycle
typical of the order Microsporida. A corn borer
larva eats a spore (the resting stage N. pyrausta);
the spore extrudes its polar filament, injecting
a sporoplasm into a midgut cell. Some devel-
opment occurs in the midgut, whereas other
sporoplasms migrate through the midgut into
the hemocoel and eventually infect other tis-
sues. In the European corn borer, infections pre-
dominantly take place in the Malpighian tubules
and in the reproductive tissues of the female in-
sect. At this point, two routes are possible for
the microsporidium (both routes occur in all
cases of infection). The infection can develop
in the midgut, causing these cells and spores to
slough into the lumen and pass from the body
in the fecal material. The fecal material then be-
comes a reservoir of N. pyrausta spores available
to infect other corn borer larvae feeding in the
same plant. This is known as horizontal trans-
mission, i.e., transmission within a generation.
This route readily occurs in nature and is an ef-
fective means of disease transmission [38]. If the
insect in which the reproductive tissues became
infected was a female, the spores either are dis-
8 Biological Control
seminated within the developing insect egg and
infect the embryo during development (transo-
varial transmission), or they are passed on the
egg shell and are consumed by the larvae at eclo-
sion (transovum transmission). Transovarial and
transovum transmission are collectively known
as vertical transmission, i.e., transmission to a
subsequent generation. Horizontal and vertical
transmission are two very effective means of
transmitting and maintaining an organism within
an ecosystem.
The impact of N. pyrausta occurs in sev-
eral ways. If the infection is intense enough (a
tremendous number of spores per insect larva),
the infected insect will die. If the insect devel-
ops to the adult with this infection, the impact of
N. pyrausta is elicited by a shortened life span,
reduction in the number of eggs laid, and a re-
duction in the number of eggs hatching and de-
veloping to maturity.
Vairimorpha necatrix is a microsporidium
that infects a great number of insects, all being
phytophagous Lepidoptera. This microsporid-
ium also can cause an acute pathology, resulting
in death of the susceptible larvae within several
hours, and is an example of an applied biolog-
ical control agent. If a newly hatched or rela-
tively early stage larva consumes an excessive
number of spores, the midgut cells are dam-
aged, allowing entry of the midgut contents into
the hemocoel, and the larva dies from bacterial
septicemia. If the larva does not die from sep-
ticemia, the spore enters the hemocoel, and an
acute infection of the fat body occurs. This infec-
tion becomes very intense, and the host usually
dies before pupation or during the pupal period.
The European corn borer is not as susceptible
to V. necatrix as are several larvae in the family
Noctuidae (cutworms, armyworms, etc.), and a
few insects develop to adults. However, because
of the intense infections at this time, the produc-
tion of offspring is unlikely. The acute pathology
caused by V. necatrix makes it improbable that
any horizontal or vertical transmission will oc-
cur. If transmission does occur, it is likely by an
insect larva feeding on infected cadavers, by can-
nibalism, or by insects feeding on foliage con-
taminated by infected insects that ruptured after
death (horizontal transmission) [39].
Apicomplexa. The gregarines and some coc-
cidia with potential as biological control agents
are representatives of this phylum. An infection
usually occurs by consumption of spores. The
development cycle involves schizogeny, which
increases the number of spores tremendously,
destroying tissues and depleting energy reserves.
This cycle takes 1 – 6 weeks, depending on the
specific organism. Mattesia grandis, a pathogen
of the boll weevil, Anthonomus grandis, is an
example of this group and contributes to over-
wintering mortality of this insect. Mattesia tro-
godermae, a pathogen of a stored-grain beetle,
Trogoderma glabrum, has great potential for
controlling these beetles in warehouses. Adult
males can be contaminated with spores and can
transmit these spores to females during copula-
tion [40].
Commercial application of pathogens with
protozoal characteristics is limited because these
organisms must be produced in vivo, in their re-
spective hosts.
2.2.3. Insect Viruses
There are two general types of insect viruses,
the occluded and nonoccluded viruses. The oc-
cluded-type viruses are characterized by hav-
ing the infective units, the virions, enclosed
by a protein capsule (occlusion body). This is
the most emphasized group relative to biologi-
cal control and includes the nuclear polyhedro-
sis viruses (NPV), the cytoplasmic polyhedro-
sis viruses (CPV), the granulosis viruses (GV),
and the entomopox viruses or insect pox viruses.
The iridescent viruses, parvoviruses and picor-
naviruses are representatives of the nonoccluded
viruses.
The occlusion body of an NPV is many-sided
and encloses several virions. The virions are rod-
shaped, and replication occurs within the cell nu-
cleus. A generalized cycle, using a nuclear poly-
hedrosis virus, is illustrated in Figure 8. The in-
clusion bodies (occlusion bodies) are consumed
by the insect and the proteinaceous capsule is
hydrolyzed in the alkaline gut, releasing the viri-
ons. The virions enter the midgut cell nucleus,
at which point they replicate and destroy the
cell, or else the virions pass through the cell
into the hemocoel and infect cell nuclei of other
tissues, replicate, encapsulate, and rupture the

Figure 7. Infection cycles of the microsporidium Nosema pyrausta
cell. Insects infected with NPV exhibit sluggish
retarded growth and a behavioral change char-
acterized by moving to the top of a host plant
where they die, and the integument usually rup-
tures. The primary tissues infected with a nuclear
polyhedrosis virus are the midgut, tracheal ma-
trix, fat body, and hypodermis. A viral infection
in these tissues with eventual rupturing of the
cells gives rise to a classical viral death where the
internal organs have liquefied, and the cuticle of
the insect is a mere “bag” of viral occlusion bod-
ies. Once this “bag” breaks, the occlusion bodies
contaminate the surface to which the insect was
attached and become available for consumption
by other susceptible insects.
Insects infected with a CPV display discol-
oration of the midgut (visible through the integu-
ment), experience retarded growth, and eventu-
ally die. The integument does not rupture. The
shape of the occlusion body varies, and several
virions, icosahedral in shape, are within each
occlusion body. The infection occurs within the
cytoplasm of the cell.
A GV infection is relatively nondescript but
is generally characterized by retarded growth of
the insect and a paling of color of the integu-
ment. In some infections, the occlusion bodies
are passed into the hemolymph, giving a milky
color. The occlusion bodies of this virus are
formed within the cell nucleus and usually con-
tain a single rod-shaped virion. Insects infected
with a pox virus are sluggish and have an ex-
tended developmental stage. The occlusion bod-
ies develop predominantly in the fat body, both
in the nucleus and in the cytoplasm of the cell.
Biological Control 9
The occlusion body contains many ovoid infec-
tive units.
Insect viruses were believed to be host spe-
cific (only infecting a single species of insect),
but research has revealed that some nuclear
polyhedrosis viruses will infect more than one
species. For example, the nuclear polyhedrosis
virus from the alfalfa looper, Autographa cali-
fornica, infects several alternate hosts [41], in-
cluding some from which a virus has never been
isolated, i.e., the European corn borer and the
black cutworm [42], [43]. This aspect of a wider
host range for a virus increases the feasibility of
using viruses in applied biological control. In-
sect viruses can be produced only in respective
hosts.
2.2.4. Fungi
Most entomopathogenic fungi belong to the
class Deuteromycetes (Fungi Imperfecti), with
some very important entomopathogenic fungi
in other classes. Insects infected with a fungus
exhibit general lethargy, slowed growth, cessa-
tion of feeding, and changes in coloration of the
integument. The difference between fungi and
other pathogens is that the former do not have
to be eaten by the insect to cause disease, but
instead grow through the insect’s skin.
A general infection cycle for fungi is illus-
trated with Beauveria bassiana as an example
(Fig. 9). The resting stage or conidium comes
in contact with the insect cuticle. The conidium
germinates and enzymatic action partly digests
the cuticle, allowing the hyphae to penetrate the
10 Biological Control

Figure 8. Infection cycle of a nuclear polyhedrosis virus

cuticle. The hyphae develop, forming a network that must be met for the fungus to be effective.
of mycelia within the body cavity. Death usu- Fungi can be produced on submerged culture or
ally occurs after this mycelial growth has spread on agar plates.
throughout the body cavity. At this time, the
mycelia grow out of the body, forming conidio-
phores and conidia. These formations are unique
and definitive in some fungi. For example, B.
bassiana conidia cover the infected insect, trans-
forming it into a white covered mummy. This
fungus is referred to as the white muscardine.
Metarrhizium anisopliae turns an insect into a
green-covered mummy and is called the green
muscardine fungus. Both these fungi infect a
wide range of insects. Nomuraea rileyi also
coats insects with a green covering but is not
as “fuzzy” as M. anisopliae. The conidia of
these fungi become airborne and eventually in- Figure 9. Infection cycle of the fungus, Beauveria bassiana
fect other insects.
Some insects exhibit a typical behavior pat-
tern once infected. Diptera (flies) infected with
Entomophthora musca climb to the top of veg- 2.3. Nematodes
etation, wrap their legs around a grass stem,
for example, and die; then mycelia grow on Nematodes (phylum Nematoda) are elongated
the external surface. A similar response occurs cylindrical worms that are parasitic in animals or
in grasshoppers infected with Entomophaga plants, or are free-living in soil or water. Nema-
grylli. These fungi forcibly discharge the coni- todes are of importance in the biological control
dia from the conidiophores. Both these fungi of insects. Several families of nematodes have
are members of the class Zygomycetes. The members parasitic to insects. Nine families show
class Oomycetes contains the species Lageni- potential as biological control agents [14]. In this
dium giganteum, a pathogen of several species article the families Mermithidae and Steinerne-
of mosquitoes. matidae are be mentioned as examples of insect
Entomopathogenic fungi are omnipresent; parasitic forms.
however, their effectiveness as a biological con-
trol agent is dependent on temperature, mois- The mermithids have both aquatic and ter-
ture, and wind. Each species has certain require- restrial species. Romanomermis culicivorax is
ments relative to these environmental variables an aquatic species that is parasitic to several

species of mosquitoes. Romanomermis culicivo-
rax hatches from an egg and actively seeks a
mosquito larva and penetrates the larval integu-
ment by use of a stylet and enzymatic secretions.
The nematode completes its development within
the insect hemocoel and then leaves the host by
boring through the integument. At this time, the
host usually dies. Once in the aquatic environ-
ment, the nematode develops to adulthood and
mates. The female deposits its eggs to continue
the cycle.
Mermis nigrescens, a representative of a
terrestrial mermithid, is mainly a parasite of
grasshoppers, but will occasionally parasitize
other insects, including beetles. Adult M. ni-
grescens live in the soil. The female climbs veg-
etation, lays her eggs, and returns to the soil.
An insect eats an egg. Once in the gut, the
egg hatches, and the nematode penetrates the
midgut, enters the hemocoel, develops within
for several weeks, and then leaves the host (the
host dies at this time); the nematode enters the
soil and develops to adulthood. Mating may
or may not occur. Reproduction can take place
parthenogenetically.
The steinernematids are represented by
Neoaplectana carpocapsae, which parasitizes
terrestrial insects within several orders. Infective
nematode larvae actively enter a host through the
mouth, the anus, or through a spiracle, and pro-
ceed to the hemocoel, where they begin devel-
opment. The steinernematids have a symbiotic
relationship with a bacterium. As the N. car-
pocapsae larvae develop, a bacterium, Achro-
mobacter nematophilus, is released through the
anus. This bacterium creates a desirable envi-
ronment in which N. carpocapsae develops. The
nematodes develop to maturity and mate, and the
female deposits her eggs. These eggs hatch, and
the larvae develop to infective stage and then
leave the host, ready to start the cycle again.
Nematodes exist in nature. However, some
nematodes, including both the aforementioned
ones, are produced commercially and, there-
fore, are available as applied biological control
agents. Methods have been developed for in vitro
production of nematodes. A high-protein source
free from bacterial contamination is a necessity
[14].
Biological Control 11
3. Biological Control of Weeds
The biological control of weeds theoretically in-
volves the use of any organism that uses the weed
as a food source or as a host. In general, most suc-
cessful programs in biological control of weeds
involve the use of phytophagous insects or in-
sects that feed on these weeds. There are several
programs throughout the world that have been
successful in controlling weeds. For illustrative
purposes, a few of the more prominent ones will
be emphasized.
3.1. Phytophagous Insects
Control of the prickly pear cactus in Australia
in the 1920s was one of the earliest successes.
Larvae of the moth Cactoblastis cactorum were
imported from Argentina and destroyed popula-
tions of this group of cacti. In the United States,
these cacti have been substantially controlled by
a cochineal insect, Dactylopius opuntiae, im-
ported from Hawaii.
The Klamath weed, Hypericum perfora-
tum, was introduced to northwestern North
America in the early 1900s and has been suc-
cessfully suppressed by two beetles, Chrysolina
hyperici and C. quadrigemina, which were im-
ported from Europe via Australia.
Musk thistle, a weed in range and pasture
land in many areas, is host to a weevil, Rhinocyl-
lus conicus, that lays its eggs in the seed head of
the thistle. Rhinocyllus conicus was introduced
into western Canada and into the United States
(Montana, Nebraska, and more recently Iowa)
and has a definite negative impact on this weed.
These three plants are examples of control
of terrestrial weeds. Successful control of an
aquatic plant, the alligator weed Alternanthera
philoxeroides, has been demonstrated in inland
waterways in Florida. Control has been obtained
with a beetle, Agasicles hygrophila, and a moth
larva, Vogtia malloi.
3.2. Plant Pathogens
The use of plant pathogens for control of weeds
is a relatively new approach. Most of the empha-
sis has been with plant pathogenic fungi. Basic
12 Biological Control
research is performed under carefully controlled
conditions to assure the specificity of pathogens.
Phytopthora palmivora is a fungus that is spe-
cific for the milkweed vine. Some of these plant
pathogenic fungi maintain themselves in the soil
and thus are excellent biological control agents.
Alternaria cassiae is a fungus that shows po-
tential for control of sicklepod. Several other
fungi are being extensively researched for bi-
ological control of weeds. This discipline will
have a tremendous impact on weed control in
the future.
3.3. Higher Animals
Tansy ragwort, Senecio jacobaea, is a biennial
weed containing pyrrolizidine alkaloids. These
alkaloids cause liver damage in livestock that
consume these weeds. However, sheep are seem-
ingly immune to this alkaloid. Therefore, they
can graze this plant and keep it from producing
seeds, thus reducing the plant population [44].
Biological control of weeds is a very active
area of research. More and more phytophagous
insects and plant pathogens are being evaluated
as potential biological control agents. Once the
basic research has been completed, several of
these organisms will most likely be used to aid
in suppression of weeds.
When selecting weedy plants as potential tar-
gets for biological control, one must be certain
that the weed is classified as a weed in all areas
of the country where it occurs. In some parts of
a country, a certain plant might be a weed. How-
ever, in other locations, the plant might be ben-
eficial as food source, habitat for wildlife, or for
ground cover. The host range of phytophagous
insects and plant pathogens also must be exten-
sively researched to be absolutely sure that de-
sirable plants that are closely related and might
become an alternative host, do not exist in the
country of introduction.
4. Biological Control of
Plant Pathogens
Plant pathogens are a group of pests that cause
tremendous loss of plant life every year and are
primarily suppressed or controlled by chemical
pesticides. However, there are biological control
agents that use these microorganisms as sources
of food or as hosts.
4.1. Macroorganisms
Fungi (smuts, rusts, and mildews) are food
sources for several insects, but applied biocon-
trol of plant pathogens by insects has not been
encouraging.
4.2. Microorganisms
The complex interrelationships between plant
pathogens and microorganisms that have the po-
tential of controlling them have been studied.
Cucumber powdery mildew has been experi-
mentally controlled by application of the fungus
Ampelomyces quisqualis [45]. Ampelomyces
quisqualis is a hyperparasite that obtains nu-
trients from the mildew. The fungus Tricho-
derma harzianum has been shown to degrade
plant pathogens, such as Schlerotium rolfsii, Rhi-
zoctonia solani, and Pythium aphanidermatum.
Plant nematodes, Criconemella xenoplax, have
been suppressed by the fungus Hirsutella rhos-
siliensis. Here again, researchers are faced with
a very complicated biological system in which
a tremendous amount of basic research must
be conducted before biological control of plant
pathogens can be realized to its full potential.
5. Other Biological Methods of
Control
5.1. Biochemicals
Interfering with an insect’s growth to make
the insect vulnerable to natural control mea-
sures and maintaining an insect in a “nonpest”
developmental stage have long been intrigu-
ing ideas for insect control. During normal in-
sect development, immature stages are regu-
lated by hormones (growth regulators). During
the adult stage, insect behavior is influenced by
pheromones and kairomones (semiochemicals).
5.1.1. Growth Regulators
Ecdysone is secreted by the prothoracic glands
and initiates cell growth. Once released into
 Biological Control 13

the hemolymph, ecdysone causes the insect to

molt. α-Ecdysone [3604-87-3], C27 H44 O6 , M r
464, mp 237 – 239 ◦ C, is a water-soluble crys-
talline compound; IR (KBr, cm−1 ) 3333, 1657;
UV (MeOH) 242 nm (ε 12400); NMR (C5 D5 N) Juvenile hormone I
[46]. It was originally isolated from silkworm

Juvenile hormone II

Juvenile hormone III

α-Ecdysone
Juvenile hormone 0

pupae. β-Ecdysone [5289-74-7], 20-hydroxy-

α-ecdysone, is a slightlymore biologically active
compound. Both α- and β-ecdysone are biosyn-
thesized from cholesterol [47]. Phytoecdysones,
steroids with molting hormone activity, have iso-Juvenile hormone 0
been isolated from several plants. The isola-
tion and synthesis of phytoecdysones and α- Juvabione [17904-27-7], a juvenoid (com-
and β-ecdysone have been described in detail pound with JH activity), was isolated from
[47]. Exogenous ecdysones will initiate molting the balsam fir tree [48]; MS 266.188 (M+
in insects when applied to the surface of the C6 H26 O3 ); UV 223 nm; IR (cm−1 ) 1722 (α, β-
cuticle. unsaturated ester), 1645 (saturated ketone).
Juvenile hormones (JHs) are synthesized in
and secreted by the corpus allatum. These hor-
mones stimulate larval development while pre-
venting development of adult characteristics.
They also promote synthesis of yolk proteins and
accumulation of these proteins by the develop-
ing oocytes. Juvenile hormones control the de-
velopment of imaginal discs. Application of JH
to a young embryo will block its development to Juvabione
a larva; however, JH may be important in normal
embryonic development of some insects. Several other juvenoids have been isolated
Five juvenile hormones, JH 0 [55333-14-7], from plants [49]. The determination of the
iso-JH 0, JHsI [13804-51-8], JH II structure of the naturally occurring JHs led
[34218-61-6], and JH III [22963-93-5], have to the synthesis of JH-analogs. Syntheses of
been isolated from insects. The JH I, II, and III JHs, juvenoids, and JH-analogs have been re-
are liquids, JH 0 and iso-JH 0 are presumed to be viewed [50], [51]. Commonly used synthetic JH-
liquids; JH I bp 130 – 145 ◦ C; IR (cm−1 ) 1715, analogs are: methoprene [40596-69-8], Altosid,
1640, 1230, 1150, 850, 1730, 1655, 1722, 1650. ZR 515, mp 135 – 136 ◦ C, d 20 0.926; hydroprene
14 Biological Control
[41096-46-2], Altozar, ZR 512; epofenonane
[57342-02-6], RO 10-3108. The applied use and
chemistry of these and other JH analogs have
been thoroughly discussed [52].
Methoprene
Hydroprene
Epofenonane
Two compounds, precocene 1 [17598-02-6]
and precocene 2 [644-06-4], isolated from the
common bedding plant, Ageratum houstoni-
anum, have anti-JH activity against several
hemipteran species [53]. Precocenes destroy the
corpora allata of insects in the orders Hemiptera,
Homoptera, and Orthoptera. There is no anti-JH
activity against holometabulous insects; how-
ever, certain ones have been forced into dia-
pause, have been sterilized, or both. Synthesis
of precocenes has been described [54].
Leaves of the neem tree, Azadirachta indica,
yield chemicals that are strong antifeedants [55],
[56]; however, these chemicals have not led to
any significant use for pest control. The main
component of the neem fractions is azadirachtin
[70357-01-6], C35 H44 O16 , a compound that acts
as a phagorepellent and as a growth regulator;
UV(MeOH) 217 nm (ε 9100); CD (MeOH), no
extrema above 220 nm; IR (KBr, cm−1 ) 3410,
(OH) 1745, 1720, 1700 (esters), 1645, 1620,
843 (double bonds); IR (chf. concentration vari-
ation) 3580 cm−1 (free OH), 3465 (intramolecu-
lar bonded OH), 3380 (intermolecular H bonded
OH).
5.1.2. Semiochemicals
Pheromones. A pheromone is a substance
that is secreted by an organism to the outside and
mediates a response in an organism of the same
species. Sex pheromones are specific chemicals
used in the mating process. Generally, they are
emitted by the female insect and attract the male
insect. Chemical structures of pheromones are
scattered among many types of volatile com-
pounds ranging from alkanes to nitrogen het-
erocycles. Earlier research usually reported one
chemical as the pheromone of an insect. Synthe-
sis of these compounds and field testing for at-
tractancy has resulted in various degrees of suc-
cess. However, although a single compound is
the major component, there are trace amounts
of additional chemicals that make up the full
complement of the pheromone [57–59].
Using open-tubular capillary chromatogra-
phy analysis, the following composition was de-
termined for the pheromone of the lesser corn-
stalk borer, Elasmopalpus lignosellus: 16.5 %
(Z)-7-tetradecen-1-ol acetate, 8.8 % (Z)-9-
tetradecen-1-ol acetate, 3.8 % 1-tetradecanol ac-
etate (C14 ), 28.2 % (Z)-9-hexadecen-1-ol ac-
etate, 17.4 % (Z)-11-hexadecen-1-ol acetate,
9.6 % 1-hexadecanol acetate (C16 ), 2.1 % (Z)-
7-tetradecen-1-ol, 2.1 % (Z)-9-tetradecen-1-ol,
7.0 % (Z)-9-hexadecen-1-ol, and 4.4 % (Z)-11-
hexadecen-1-ol [59]. The organic synthesis of
sex pheromones has been reviewed [50], [60],
[61]. Biosynthesis of pheromones takes place in
a glandular area of the abdominal tip of the adult
female and is regulated by a hormonal factor pro-
duced in the brain [62].
Several pheromones have been synthesized
and these synthetic pheromones are available for
certain pests of agricultural crops [63].
 Biological Control 15

Kairomones. Kairomones are compounds and field tests against several insect pests, phy-
given off by one species that induce a greater be- tophagous mites, and a plant parasitic nematode.
havioral response in another species than in the
species exuding the compound. Kairomones in-
clude host-seeking chemicals, e.g., compounds
that aid parasitoids in search of suitable hosts.
Host-seeking compounds, extracted from the
wings (scales) of Heliothis zea and Cadra
cautella, increase parasitization of eggs by
Trichogramma evanescens. These compounds
stimulate the parasitoid to search for suitable
host eggs.
Some plants produce substances that attract
insects, e.g., cucurbitacins. Cucurbitacins are
oxygenated tetracyclic triterpenes produced by Avermectin R1 R2 R3 CAS-Reg.
No.
plants of the family Cucurbitaceae, i.e., Cucur-
bitacin B [6199-67-3]. A complete discussion A1 a C 2 H5 CH3 [65195-51-9]
A1 b CH3 CH3 [65195-52-0]
of the chemistry of these compoundsis available A2 a OH C2 H5 CH3 [65195-53-1]
[64]. A2 b OH CH3 CH3 [65195-54-2]
B1 a C 2 H5 H [65195-55-3]
B1 b CH3 H [65195-56-4]
B2 a OH C2 H5 H [65195-57-5]
B2 b OH CH3 H [65195-58-6]

Where R1 is absent, the double bond (=) is present. Both sugars

are α-l-olendrose.

The fruits of wild species of Cucurbita con- 5.2. Autocidal

tain amounts of these chemicals that are toxic
to animals. Cucurbitacins are also present in
extremely small quantities, < 0.02 mg/g, in do-
mesticated fruits of C. ficifolia, C. maxima, C.
mixtra, C. moschata, and C. pepo [65]. This
small amount, however, acts as an arrestant
(kairomone) and feeding stimulant to several
beetles in the genus Diabrotica [66].
Autocidal methods involve alteration of genetic
material of insects, thereby reducing the insect’s
ability to reproduce. Briefly, after the genetic
material of an insect population is altered, this
population is released into nature. Mating of the
genetically abnormal population with the wild
population causes reduction in the natural pop-
ulation.

5.1.3. Bioderived Toxicants 5.2.1. Sterile Male

Some organisms produce chemicals that are
mere poisons without any novel mode of ac-
tion, such as an attractant and/or growth regu-
lator. The avermectins are a class of compounds
described as pentacyclic lactones isolated from
fermentation of the actinomycete Streptomyces
avermitilis. These natural products contain an α-
l-oleandrosyl-α-l-oleandrosyl disaccharide at-
tached to the macrocyclic lactone ring through
the allylic C13 -hydroxy group [67]. These com-
pounds have demonstrated toxicity in laboratory
The primary autocidal method is the sterile
male technique in which tremendous numbers
of male insects are sterilized either by radiation
or by chemical means, and released into natural
populations in such numbers that the sterilized
males greatly outnumber the naturally occurring
ones. This technique has been employed with
success against the screwworm, Cochliomyia
hominivorax, Mediterranean fruit fly, Ceratitis
capitata, melon fly, Dacus cucurbitae, Mexican
fruit fly, Anastrepha ludens, and pink bollworm,
Pectinophora gossypiella.
16 Biological Control
5.2.2. Hybrid Sterility
Hybrid sterility involves the crossing of two
closely related but different species of insects
with production of sterile progeny and subse-
quent release in natural populations. These hy-
brids must be fully viable and able to compete
with feral adults. Other genetic alterations are:
(1) cytoplasmic incompatibility – certain strains
within a species when crossed produce either
sterile offspring, or sterile and fertile offspring,
or sterile males; (2) conditional lethals – certain
alleles of genes controlling adaptability produce
normal effects at certain temperature ranges but
produce lethal effects at higher or lower temper-
atures; (3) growth alterations – for example, in-
troducing a gene for “nondiapause” into a strain
of insects that must diapause in order to survive
the winter.
6. Formulation, Application, and
Efficacy
Insect Control. Formulation and applica-
tion of biological control agents are a “hand-in-
glove” type of relationship. The proposed use
of an agent dictates to a great extent the way in
which that agent must be formulated to elicit its
greatest possible impact. Formulation must be
done with the recognition that biological con-
trol agents are living organisms which vary in
their susceptibilities to adverse environmental
conditions.
The formulation techniques for parasitoids
and predators are limited. Basically, these in-
sects are colonized, produced in large numbers
in an insectary, and then released into the en-
vironment at a time predetermined to be most
advantagous to the beneficial insect.
A special case of formulation and application
of parasitoids is employed when Trichogramma
are used in applied biological control. Briefly,
Sitotroga cereallella or Anagasta kuehniella
adults are constrained to lay many eggs on sheets
of cardboard; these eggs are parasitized by Tri-
chogramma adults, and the cards of parasitized
eggs are placed in an ecosystem containing re-
cently oviposited eggs of a pest insect. This pro-
cedure is timed so that the Trichogramma will
emerge within a few hours after being placed in
the environment. Trichogramma maidis is uti-
lized in eastern France to control populations of
the European corn borer in corn. Researchers ob-
tained 90 – 95 % control by placing 300000 para-
sitized Anagasta kuehniella eggs/ha [68] in corn
fields infested with the European corn borer.
Techniques also have been developed to apply
Trichogramma with a crop-dusting plane. The
host eggs are sometimes oviposited on a car-
rier, such as wheat bran, and this admixture is
air – dropped over an insect-infested crop area.
Pediobius foveolatus, a parasitoid of the Mex-
ican bean beetle, was released annually in Mary-
land. Approximately 490000 parasitoids were
released over 94000 ha of soybeans, resulting in
up to 84 % parasitism of the Mexican bean beetle
[69].
Excellent control of aphids on potatoes was
obtained by releasing a lacewing, Chrysopa
spp., and a coccinellid, Coccinella septempunc-
tata, at the same time. These researchers re-
leased 210300 and 74600/ha of Chrysopa and
C. septempunctata, respectively [70].
Pathogens are formulated in the same way
that chemical insecticides are, i.e., as dusts, wet-
table powders, flowable concentrates, granules,
or baits. An insect-pathogen dust or granule is
made by mixing the pathogen with an inert mate-
rial called a diluent. Diluents most often used are
talcs, prophyllite clays, clays, calcium carbon-
ate, precipitated hydrated silicates, silicon diox-
ides, synthetic calcium silicate, and diatoma-
ceous earth. Potato starch, cornstarch, wheat
flour, wheat bran, ground corn cobs, and ground
corn have been used as experimental diluents.
The basic differences between a dust and a gran-
ule is the mere size of the diluent.
Wettable powders are formulations of
pathogens containing sufficient wetting agent
that the pathogen will be wetted and suspendible
in water. Powders can be any of the named dilu-
ents, only finely ground, so that a sprayable end
product is obtained. A flowable concentrate is
essentially a wettable powder suspended in an
oil or liquid base of some type that can readily
be mixed with water to form a stable suspension.
Baits usually are botanical products, such as
apple pomace, grape pomace, citrus pulp, wheat
bran, etc., that are food sources and attract the
target insect.
In all these formulations, great care is taken to
assure shelf life, stability in the field, and mainte-

nance of viability of the pathogen. Commercial
additives are available to protect the pathogen
from severe drying or wetting and from ultravi-
olet light.
Wettable powder and flowable concentrate
formulations are employed when complete cov-
erage of an area or vegetation is necessary
for maximum efficiency. The use of Bacil-
lus thuringiensis subspecies israelensis for
mosquito and black fly control is a situation in
which these formulations are used.
Two wettable powder formulations of B.
thuringiensis subspecies israelensis, Bactimos
(Biochem Products, USA) and Vetobac (Ab-
bott Laboratories, USA), and a flowable concen-
trate, Teknar (Sandoz, USA), were tested against
mosquito populations in central Italy [71]. Bac-
timos and Teknar were applied at 0.5 kg/ha and
2.5 L/ha, respectively, to control the mosquito
Aedes caspius in a salt marsh. These applications
gave complete control of the larvae. Vectobac at
0.5 kg/ha gave a range of 67 – 91 % control. A
flowable concentrate formulation of Teknar was
prepared to give a concentration of 10 ppm and
applied to the width of a fresh water stream for
1 min with complete elimination of the black fly
larvae [72].
Aqueous suspensions of insect viruses are
prepared from wettable powder formulations.
Viruses are quantified as polyhedral inclusion
bodies (PIB) per unit mass. The European pine
sawfly, Neodiprion sertifer, can be controlled on
Pinus resinosa with an application of 2.5×109
PIB/ha [73]. Application rates of this magnitude
have been used to control other forest pests [74]
and pests of other agricultural crops, i.e., cole
crops, cotton, and soybeans [75]. Granular for-
mulations are used in an ecosystem for place-
ment of a pathogen on a plant or within a certain
area, or for incorporation into soil. For exam-
ple, larvae of the European corn borer feed on a
corn plant at the interface where the developing
leaves are unfolding. Because of the morphol-
ogy of the plant, there is a natural funnel that
will direct granules of a pathogen to this area and
thus make them immediately available for con-
sumption by the insect. Not only is the pathogen
more readily available to the insect, but also it is
protected from ultraviolet radiation.
An 11.25 wt % formulation of Thuricide
HPC on clay granules was applied at 1.12 kg
of B. thuringiensis HPC/ha to control the Eu-
Biological Control 17
ropean corn borer on corn [76]. This applica-
tion caused a significant reduction in damage
by the borer compared to the untreated check.
Several mosquito habitats are in areas of heavy
vegetation. A granular formulation, when ap-
plied in this type of environment, will pass
through the vegetation, enter the water, and be-
come available to the mosquito larvae. Granu-
lar formulations of Bacillus popilliae are incor-
porated into soil for control of Japanese bee-
tles in Russia [77]. A specialized formulation
and application is the impregnation of founda-
tion wax with Bacillus thuringiensis subspecies
galleriae for control of greater wax moth lar-
vae in beehives [78]. Nosema locustae is for-
mulated as a bait on wheat bran and applied to
rangeland to control grasshoppers at 2.5×109
N. locustae spores on 2 kg of wheat bran/ha
[79]. This application reduced the population
of immature grasshoppers by 50 %. A simi-
lar treatment effectively reduced populations of
the Mormon cricket, Anabrus simplex [80]. Ne-
matodes, Neoaplectana carpocapsae, applied as
an aqueous suspension (
1000 nematodes/mL)
were highly effective against larvae of the arti-
choke plum moth, Platytilia cardiudactyla [81].
The neoaplectanid nematodes show potential
for control of several insect pests [82]. Ro-
manomermis culicivorax, a nematode parasite
of mosquitoes, has been tested worldwide for
mosquito control. Different stages of the nema-
tode, egg, pre-, and postparasitic, are applied to
the aqueous environment at rates of a few hun-
dred to 200000 units/m2 with varying degrees
of success. A wettable powder or dust formula-
tion of Hirsutella thompsoni, a myoacaricide,
was effective in stimulating premature fungal
epizootics in citrus rust mite populations [83].
The environs of a greenhouse create special
pest problems but also present a controllable sit-
uation for the use of biological control agents.
The successes, problems, and failures of biolog-
ical control in such ecosystems have been re-
viewed [84], [85]. In a greenhouse-like situation,
the spotted spider mite, Tetranychus urticae, was
controlled in a plastic walk-in tunnel by intro-
ducing one predatory mite per strawberry plant
[86]. Control of the greenhouse whitefly, Tri-
aleuroes vaporariorum, is difficult. A combina-
tion of attracting the whitefly to yellow sticky
traps and using the parasitoid Encarsia formosa
has resulted in 90 % parasitization of this pest on
18 Biological Control
spring tomatoes in the greenhouse [87]. In other
research [88], several species of parasitoids with
the potential of controlling the chrysanthemum
leaf miner have been identified.
The use of fishes to control insects is pri-
marily focused on controlling the larvae of
mosquitoes that transmit human disease. Gam-
busia affinis, a larvivorous fish, is a native of
the southeastern United States but has spread to
most parts of the world where it can survive and
give excellent mosquito control. Gambusia affi-
nis seeded at 2500 – 5000 fish/ha will reproduce
quickly and provide complete mosquito control
in 6 – 8 weeks [89], [90].
Spiders definitely have an impact on control
of insects; however, quantification is difficult
[91].
Weed and Plant Pathogen Control. My-
coherbicides are also formulated as an aqueous
suspension and sprayed on foliage for control
of weeds. Spores of Colletotrichum coccodes,
a fungus, were sprayed on velvetleaf, Abutilon
theophrasti, at 107 and 109 spores/m2 in 467.5,
935, and 1870 L of H2 O/ha. These concentra-
tions of spores reduced the velvetleaf biomass by
40 and 50 % at the 107 and 109 treatment levels,
respectively [92]. Progress has been made us-
ing a rust, Puccinia chondrilla, to suppress rush
skeletonweed, Chondrilla juncea [93]. Plant
size, number of flowers, and seed viability have
been reduced, but there has been no substantial
reduction in plant populations.
Damage caused by the root-knot nematode
Meliodogyne javanica has been reduced using
biological control. Tomato roots containing the
nematode infected with Bacillus penetrans were
air-dried, finely ground, and incorporated into
the soil at 212 – 600 mg/kg, reducing the num-
ber of nematodes and the damage caused by
the nematodes [94]. Organic extracts from the
fungus Aphanocladium album induce teliospore
formation on rust fungi. Puccinia graminis, P.
dispersa, P. striiformis, and P. sorghi formed
premature telia on plants treated with the ex-
tract [95]. The biology of introduced predators
and parasitoids of arthropod and weed pests is
available [96].
Other Methods of Biological Control.
Growth regulators have been tested for insect
suppression. Lethal effects were induced in
a population of western spruce budworms at
dosages comparable to several conventional in-
secticides [97]. Treating male or female adults
of the western spruce budworm with ZR575,
ZR512, or RO 10-3108 reduced the reproduc-
tive potential of these insects [98]. Precocene 1
and 2 deterred Heliothis zea larvae from feed-
ing in laboratory studies, resulting in starvation
[99].
Azadirachtin from the neem tree interfered
with molting of the face fly, Musca autumnalis,
when larvae were exposed to concentrations as
low as 0.00001 µg/mL [100]. The application of
2 µg of azadirachtin/larva killed 92 – 100 % of
Japanese beetle, Popillia japonica [101].
Insect pheromones have potential for sev-
eral uses in insect management, e.g., monitoring
populations, trapping insects to a point source,
and interfering with mating. The science of in-
sect pheromones involves several steps, from
isolation, identification, and synthesis of the
pheromone to learning the behavioral change
that these chemicals manifest in a target in-
sect. The state of the art of pheromones has
been reviewed [102]. Minute amounts of the
pheromone of a given species of insect will at-
tract the male. In some insects, several com-
pounds constitute the pheromone. In H. zea,
maximum male response is elicited with com-
pounds (Z)-7-hexadecenal, (Z)-9-hexadecenal,
(Z)-11-hexadecenal, and hexadecanal, whereas
H. virescens males respond to a mixture of
the above chemicals plus (Z)-9-tetradecenal,
tetradecanal, and (Z)- 11-hexadecen-1-ol [57],
[58]. The female sex pheromone of the lesser
cornstalk borer, Elasmopalpus lignosellus, is
composed of 10 compounds (Section 5.1.2). A
mixture of four of these compounds, 91.6 µg of
(Z)-11-hexadecen-1-ol acetate, 11.2 µg of (Z)-
9-tetradecen-1-ol, 46.4 µg of (Z)-9-tetradecen-
1-ol acetate, and 86.8 µg of (Z)-7-tetradecen-1-
ol acetate applied to a rubber septum produced
maximal capture of lesser cornstalk borer fe-
males [59].
Cucurbitacins act as kairomones to beetles in
the subtribe Diabroticites, which includes sev-
eral species that feed on Cucurbitaceae. This
tribe also includes the corn rootworm beetles.
Amounts as small as 0.003 µg cause complete
arrest of these beetles. If they pass by a point
source, they stop and begin compulsive feed-
ing until the food source is depleted [66]. Re-

searchers have demonstrated a strong correlation
between total Cucurbitacin content and the ex-
tent of aggregation and feeding by two species of
Diabrotica. Other researchers have proposed us-
ing fruits of Cucurbitaceae coated with a chem-
ical insecticide to bait insects to a point source
and kill them [103].
Bioderived Toxicants. Ivermectin [70288-
86-7], 22,23-dihydro derivative of avermectin,
is active at extremelylow dosages against a wide
variety of arthropod parasites and nematodes
[104]; i.e., 0.001 mg/kg is effective against im-
mature stages of Dirofilaria immitis, when ad-
ministered orally to dogs. Also, 0.02 mg/kg is
efficacious against Haematopenus suis when in-
jected subcutaneously in hogs. A subcutaneous
dosage of 200 µg/kg killed blood sucking flies
on steers [105]. Pheromone production was sig-
nificantly reduced when adult boll weevils, An-
thonomus grandis were topically treated with
7 µg of avermectin B1 [106].
The sterile autocidal male technique has been
used successfully but depends on the ability to
rear large numbers of insects for sterilization
and release. The screwworm, Cochliomyia ho-
minivorax, has been eliminated from Curaçao
and southeastern United States. A program is
ongoing to suppress the screwworm in Mexico
and inhibit its spread to the southwestern United
States. To accomplish this, ca. 350×106 sterile
flies are released weekly. The established popu-
lation of flies has been pushed back in excess of
600 km from the United States – Mexican bor-
der.
The sterile insect technique is a major compo-
nent of the Mediterranean fruit fly, Ceratitis cap-
itata, program in California. Up to 40×106 flies
are released weekly, enabling success in keeping
this pest out of California.
An extensive program has been undertaken
to suppress populations of the codling moth,
Laspeyresia pomonella, in the state of Wash-
ington, United States, and British Columbia,
Canada. The codling moth is widely distributed
and feral populations continually migrate into
suppression areas rending the program ineffec-
tive.
Other autocidal techniques are generally on
an experimental basis. The hybrid sterility tech-
nique crossing Heliothis virescens males with
H. subflexa females has been tried experimen-
Biological Control 19
tally in Puerto Rico, indicating a good potential
for infusion of the sterility factor into the native
population [107].
Whenever a component is used to manage a
pest population, the question of safety arises.
Usually, pest control is undertaken to protect a
commodity that is in the food chain which even-
tually is consumed by humans. Thus, human
safety is extremely important. Recently, safety
to the environment and to the ecological bal-
ance has become recognized as important. The
organisms and chemicals discussed in this article
are entities that are indigenous to a certain eco-
logical niche or they are synthesized chemical
compounds that are an instrumental part of na-
ture. Thus, if these components are used within
the environmental framework from which they
evolved, safety is relatively assured.
Exhaustive lists are available for some organ-
isms, stating which target and nontarget organ-
isms are susceptible [108]. For others, guidelines
for safety tests and registration have been estab-
lished [109], [110]. In depth studies have been
conducted examining the effect of insect nuclear
polyhedrosis viruses in mammalian cells [111],
with no consistent deleterious effects directly at-
tributable to the viruses being observed.
Prior to inoculative releases, phytophagous
insects and plant pathogens that are candidate
biological control agents are subject to exhaus-
tive host range studies to be certain that the in-
troduced species will not become pests in a dif-
ferent ecosystem.
In general, the biological control agents dis-
cussed here will not have a detrimental impact
when used sensibly in an integrated pest man-
agement program.
7. References
1. C. B. Huffaker, P. S. Messenger (ed.): Theory
and Practice of Biological Control, Academic
Press, New York 1976.
2. P. Debach: Biological Control by Natural
Enemies, Cambridge University Press, New
York 1974.
3. C. P. Clausen: Entomophagous Insects,
McGraw-Hill, New York 1940.
4. E. Kurstak (ed.): Microbial and Viral
Pesticides, Marcel Dekker, New York 1982.
5. G. E. Cantwell (ed.): Insect Diseases, vol. 1
and 2, Marcel Dekker, New York 1974.
20 Biological Control
6. E. A. Steinhaus (ed.): Insect Pathology: An
Advanced Treatise, vol. 1 and 2, Academic
Press, New York 1963.
7. E. W. Davidson (ed.): Pathogenesis of
Invertebrate Microbial Diseases, Allanhead,
Osmun Publ., Totawa, N.J., 1982.
8. Ullmann, 4th ed., 13 : 238.
9. H. D. Burges, N. W. Hussey (ed.): Microbial
Control of Insects and Mites, Academic Press,
New York 1971.
10. H. D. Burges (ed.): Microbial Control of Pests
and Plant Diseases 1970 – 1980, Academic
Press, New York 1981.
11. L. A. Bulla, Jr., T. C. Cheng (ed.): Comparative
Pathobiology, Biology of the Microsporidia,
vol. 1, Plenum Publishing, New York 1976.
12. K. M. Smith: Virus-Insect Relationships,
Longman Group Ltd., London 1976.
13. D. C. Kelly, J. Gen. Virol. 63 (1982) 13.
14. G. O. Poinar, Jr.: Nematodes for Biological
Control of Insects, CRC Press, Boca Raton,
Fla., 1979.
15. J. J. Menn, M. Beroza (ed.): Insect Juvenile
Hormones: Chemistry and Action, Academic
Press, New York 1972.
16. D. A. Nordlund, R. L. Jones, W. J. Lewis (ed.):
Semiochemicals: Their Role in Pest Control, J.
Wiley & Sons, New York 1981.
17. M. Beroza (ed.): Pest Management with Insect
Sex Attractants and Other
Behavior-Controlling Chemicals, Amer.
Chem. Soc., Washington, D.C., 1976.
18. V. B. Wigglesworth: Insect Hormones, W. H.
Freeman & Co., San Francisco 1970.
19. Biological Control of Pests in China, USDA,
Office of International Cooperation and
Development, Scientific and Technical
Exchange Division, China Program,
Washington, D.C., 1982.
20. R. J. Cook, K. F. Baker: The Nature and
Practice of Biological Control of Plant
Pathogens, The American Phytopathology
Society, St. Paul 1983.
21. R. Van den Bosch, P. S. Messenger, A. P.
Gutierrez: An Introduction to Biological
Control, Plenum Publishing, New York 1982.
22. R. M. Anderson, E. U. Canning, A. E. R.
Taylor, R. Muller (ed.): “Parasites as
Biological Control Agents,” Symposia of the
British Society for Parasitology, vol. 19,
Cambridge University Press, Cambridge 1982.
23. H. G. Miltenburger (ed.): Safety Aspects of
Baculoviruses as Biological Insecticides,
Bundesministerium für Forschung und
Technologie, Bonn 1980.
24. Reference 1, F. J. Simmonds, J. M. Franz, R. I.
Sailer: “History of Biological Control,”
Chap. 2.
25. W. A. Baker, W. G. Bradley, C. A. Clark:
Biological Control of the European Corn
Borer in the United States, USDA Technical
Bulletin No. 983, 1948, pp. 180 – 181.
26. T. A. Angus, Nature (London) 173 (1954)
545 – 546.
27. Bioferm Corp., US 3073749, 1963 (J. C.
Megna).
28. Reference 6, A. M. Heimpel, T. A. Angus:
“Diseases Caused by Certain Sporeforming
Bacteria,” vol. 2, Chap. 2.
29. M. B. Mohd-Salleh, L. C. Lewis, J. Invertebr.
Pathol. 39 (1982) 290 – 297.
30. H. deBarjac, A. Bonnefoi, Entomophaga 7
(1962) 5 – 31.
31. H. deBarjac, A. Bonnefoi, Entomophaga 18
(1973) 5 – 17.
32. J. Krywienczyk, H. T. Dulmage, P. G. Fast, J.
Invertebr. Pathol. 31 (1978) 372 – 375.
33. L. J. Goldberg, J. Margalit, Mosq. News 37
(1977) 355 – 358.
34. J. Farkas, K. Sebesta, K. Horska, Z. Samek, et
al., Collect. Czech. Chem. Commun. 34
(1969) 1118 – 1119.
35. M. B. Mohd-Salleh, C. C. Beegle, L. C. Lewis,
J. Invertebr. Pathol. 35 (1980) 75 – 83.
36. International Mineral & Chemicals Corp., US
3758383, 1973 (T. R. Shieh, M. H. Rogoff).
37. N. D. Levine, J. O. Corliss, F. E. G. Cox, G.
Deroux et al., J. Protozool. 27 (1980) 37 – 58.
38. L. C. Lewis, Can. Entomol. 110 (1978)
897 – 900.
39. L. C. Lewis, R. D. Gunnarson, J. E. Cossentine,
Can. Entomol. 114 (1982) 599 – 603.
40. T. J. Shapas, W. E. Burkholder, G. M. Boush, J.
Econ. Entomol. 70 (1977) 469 – 474.
41. P. V. Vail, D. L. Jay, J. Invertebr. Pathol. 21
(1973) 198 – 204.
42. L. C. Lewis, R. E. Lynch, J. J. Jackson,
Environ. Entomol. 6 (1977) 535 – 538.
43. L. C. Lewis, J. R. Adams, J. Invertebr. Pathol.
33 (1979) 253 – 256.
44. S. H. Sharrow, W. D. Mosher, J. Range
Manage. 35 (1982) 480 – 482.
45. L. Sundheim, Plant Pathol. 31 (1982)
209 – 214.
46. J. B. Siddall, A. D. Cross, J. H. Fried, J. Am.
Chem. Soc. 88 (1966) 862 – 863.
47. K. Nakanishi: “Steroids,” in K. Nakanishi, T.
Goto, S. Ito, S. Natori et al.: Natural Products,
Chemistry, vol. 1, Academic Press, New
York-London 1974, p. 535.

48. W. S. Bowers, H. M. Fales, M. J. Thompson,
E. C. Uebel, Science 154 (1966) 1020 – 1021.
49. N. Wakabayashi, R. M. Waters: “Juvenile
Hormones and Analogs,” in E. D. Morgan, B.
Mandava (ed.): Handbook on Natural
Pesticides: Insects, vol. 1, CRC Press, Boca
Raton, Fla., (in press).
50. K. Mori: “Synthetic Chemistry of Insect
Pheromones and Juvenile Hormones,” in
Recent Developments in the Chemistry of
Natural Carbon Compounds, vol. 9,
Publishing House of the Hungarian Academy
of Sciences, Budapest 1979, pp. 11 – 209.
51. Reference 47, S. Nozoe: “Mono- and
Sesquiterpenes,” p. 39.
52. C. A. Hendrick: “Juvenile Hormone Analogs:
Structure-Activity Relationships,” in J. Coats
(ed.): Insecticide Mode of Action, Academic
Press, New York 1982, pp. 315 – 402.
53. W. S. Bowers, T. Ohta, J. S. Cleere, P. A.
Marsella, Science 193 (1976) 542 – 547.
54. W. S. Bowers: “Toxicology of Precocene”, in
J. Coats (ed.): Insecticide Mode of Action,
Academic Press, New York 1982, 403 – 427.
55. A. Juss: “Natural Pesticides from the Neem
Tree (Azadirachta indica),” in H. Schmutterer,
K. R. S. Ascher, H. Rembold (ed.):
Proceedings of the First International Neem
Conference, German Agency for Technical
Cooperation, Eschborn, Federal Republic of
Germany, 1981.
56. J. D. Warthen, Jr., USDA, Sci. and Educ.
Admin., Agricultural Research Results, 1979,
no. 1.
57. J. A. Klun, J. R. Plimmer, B. A.
Bierl-Leonhardt, A. N. Sparks, et al., Science
204 (1979) 1328 – 1330.
58. J. A. Klun, J. R. Plimmer, B. A.
Bierl-Leonhardt, A. N. Sparks, J. Chem. Ecol.
6 (1980) 165 – 175, 177 – 183.
59. R. E. Lynch, J. A. Klun, B. A. Leonhardt, M.
Schwarz, Environ. Entomol. 13 (1984)
121 – 126.
60. J. M. Brand, J. C. Young, R. M. Silverstein:
“Insect Pheromones: A Critical Review of
Recent Advances in Their Chemistry, Biology,
and Application,” Prog. Chem. Org. Nat’l.
Prod. 37 (1979) 1 – 190.
61. H. J. Bestman, “Stereoselective Synthesis of
Pheromones via Phosphonium Ylides,” in P.
Doyle, T. Fujita (ed.): Pesticide Chemistry:
Human Welfare and the Environment,
Synthesis and Structure Activity Relationships,
vol. 1, Pergamon Press, Oxford 1982,
pp. 77 – 82.
Biological Control 21
62. A. K. Raina, J. A. Klun, Science 225 (1984)
531 –532.
63. W. Klassen, R. L. Ridgway, M. Inscoe:
“Chemical Attractants in Integrated Pest
Management Programs,” in A. F. Kydonieus,
M. Beroza (ed.): Insect Suppression with
Controlled Release Pheromone Systems, vol. 1,
CRC Press, Boca Raton, Fla., 1982.
64. D. Lavie, E. Glotter: “The Cucurbitanes, a
Group of Tetracyclic Triterpenes,” in Progress
in the Chemistry of Organic Products, vol. 29,
Springer-Verlag, Wien, New York 1971,
pp. 308 – 362.
65. R. L. Metcalf, A. M. Rhodes, R. A. Metcalf, J.
Ferguson et al., Environ. Entomol. 11 (1982)
931 –937.
66. R. L. Metcalf, Bull. Entomol. Soc. Am. 25
(1979) 30 – 35.
67. R. A. Dybas: “Avermectins: Their Chemistry
and Pesticidal Activities,” in P. Doyle, T. Fujita
(ed.): Pesticide Chemistry: Human Welfare
and the Environment, Synthesis and Structure
Activity Relationships, vol. 1, Pergamon Press,
Oxford 1982, pp.83 – 90.
68. M. Stengel, Entomophaga 27 (no◦ H. S.)
(1982) 105 – 114.
69. L. M. Stevens, A. I. Steinhauer, J. R. Coulson,
Environ. Entomol. 4 (1975) 947 – 952.
70. W. A. Shands, G. W. Simpson, R. H. Storch, J.
Econ. Entomol. 65 (1972) 799 – 805.
71. G. Majori, A. Ali, J. Invertebr. Pathol. 43
(1984) 316 – 323.
72. L. A. Pistrang, J. F. Burger, Can. Entomol. 116
(1984) 975 – 981.
73. J. D. Podgwaite, P. Rush, D. Hall, G. S. Walton,
J. Econ. Entomol. 77 (1984) 525 – 528.
74. Reference 4, J. C. Cunningham, pp. 335 – 386.
75. Reference 4, W. C. Yearian, S. Y. Young, pp.
387 – 423.
76. R. E. Lynch, L. C. Lewis, E. C. Berry, J. Econ.
Entomol. 73 (1980) 4 – 7.
77. C. Bajan, T. Bilewicz-Pawinska, A. Fedorka,
K. Kmitova: Report 8th International Plant
Protection Congress, Sec. 5, Moscow 1975,
pp. 33 – 40.
78. H. D. Burges, J. Invertebr. Pathol. 28 (1976)
217 – 222.
79. J. E. Henry, Ann. Rev. Entomol. 26 (1981)
49 – 73.
80. J. E. Henry, J. A. Onsager, Entomophaga 27
(1982) 197 – 201.
81. M. A. Bari, H. K. Kaya, J. Econ. Entomol. 77
(1984) 225 – 229.
82. R. Gaugler, J. Nematol. 13 (1981) 241 – 249.
22 Biological Control
83. C. W. McCoy, T. L. Couch, Fla. Entomol. 65
(1982) 116 – 126.
84. N. W. Hussey, L. Bravenboer: “Control of
Pests in Glasshouse Culture by the
Introduction of Natural Enemies,” in C. B.
Huffaker (ed.): Biological Control, Plenum
Publishing, New York 1971, pp. 195 – 216.
85. I. J. Wyatt: “Progress Towards Biological
Control Under Glass,” in D. P. Jones, M. E.
Solomon (ed.): Biology in Pest and Disease
Control, Blackwell Scientific Publ., London
1974, pp. 293 – 301.
86. J. V. Cross, Plant Pathol. 33 (1984) 417 – 423.
87. M. Van De Veire, V. Vacante, Entomophaga
29 (1984) 303 – 310.
88. S. J. Cornelius, H. C. J. Godfray, Entomophaga
29 (1984) 341 – 345.
89. R. Haas, R. Pal, Bull. Entomol. Soc. Am. 30
(1984) 17 – 25.
90. E. C. Bay, C. O. Berg, H. C. Chapman, E. F.
Legner in C. B. Huffaker, P. S. Messenger
(ed.): Theory and Practice of Biological
Control, Academic Press, New York 1976,
Chapter 18.
91. S. E. Riechert: “Spiders as Biological Control
Agents,” Ann. Rev. Entomol. 29 (1984)
299 – 320.
92. A. R. Gotlieb, M. H. Brosseau, A. K. Watson:
Abstracts of 1984 Weed Science of America
Meeting, Miami, Fla., 1984, p. 68.
93. E. B. Adams, R. F. Line, Phytopathology 74
(1984) 745 – 748.
94. G. R. Stirling, Phytopathology 74 (1984)
55 – 60.
95. H. R. Forrer, J. Werder: “The Influence of
Antagonistic Fungi on the Spore-Formation of
Rust Fungi,” in H. Geissbühler (ed.): Advances
in Pesticide Science, Pergamon Press, Oxford
1978, Part 2, pp. 383 – 388.
96. C. P. Clausen (ed.): “Introduced Parasites and
Predators of Arthropod Pests and Weeds: A
World Review,” Agriculture Handbook
No. 480, U.S. Agriculture Research Service,
USDA, Washington, D.C., 1978.
97. J. L. Robertson, R. A. Kimball, Can. Entomol.
111 (1979) 1361 – 1368.
98. J. L. Robertson, R. A. Kimball, Can. Entomol.
111 (1979) 1369 – 1380.
99. C. S. Wisdon, J. T. Smiley, E. Rodriguez, J.
Econ. Entomol. 76 (1983) 993 – 998.
100. I. A. Gaaboub, D. K. Hayes, Environ. Entomol.
13 (1984) 803 – 812.
101. T. L. Ladd, Jr., J. D. Warthen, Jr., M. G. Klein,
J. Econ. Entomol. 77 (1984) 903 – 905.
102. J. A. Klun: “Insect Sex Pheromones,” in J. L.
Hilton (ed.): Agricultural Chemicals of the
Future, Proc. Beltsville Agric. Symp. 8,
Rowman & Allanheld, Totowa, N.J., 1985,
pp. 381 – 386.
103. A. M. Rhodes, R. L. Metcalf, E. R. Metcalf, J.
Am. Soc. Hortic. Sci. 105 (1980) 838 – 842.
104. W. C. Campbell, M. H. Fisher, E. O. Stapley,
G. Alberts-Schönberg et al., Science 221
(1983) 823 – 828.
105. H. A. Standfast, M. J. Muller, D. D. Wilson,
J. Econ. Entomol. 77 (1984) 419 – 421.
106. J. E. Wright, J. Econ. Entomol. 77 (1984)
1029 – 1032.
107. D. F. Martin, M. L. Laster, F. I. Proshold, P. D.
Lindgren et al., Environ. Entomol. 13 (1984)
701 – 707.
108. C. C. Beegle, L. C. Lewis, T. Yamamoto:
“Bacteria,” in C. M. Ignoffo (ed.): Handbook
of Naturally Occurring Pesticides: Microbial
Insecticides, CRC Press, Boca Raton, Fla.,
1985.
109. H. D. Burges, J. Huber, G. Croizier,
Entomophaga 25 (1980) 341 – 348.
110. H. D. Burges, A. Krieg, P. Luthy, H. deBarjac,
Entomophaga 27 (1982) 225 – 236.
111. R. Reimann, H. G. Miltenburger,
Entomophaga 27 (1982) 267 – 276.