Chapter 1
Entomopathogens Used as Microbial
Control Agents
L.A. Lacey
IP Consulting International, Yakima, WA, United States
1.1 INTRODUCTION TO
ENTOMOPATHOGENS USED IN THE
MICROBIAL CONTROL OF INSECT
AND MITE PESTS
The goal of this book is to provide a current, comprehen-
sive, and enduring reference for microbial control and the
underlying principles of insect pathology that determine its
success. Central to this must be a clear understanding of the
nature and ecology of the microbes that cause diseases in
pest species.
1.1.1 A Brief History of Entomopathogens
and Their Use as Microbial Control
Agents
Diseases of honeybees and silkworms have been rec-
ognized for millennia (Davidson, 2012), but the causal
agents of insect diseases were not demonstrated until
1835, when Augusto Bassi associated a disease of the silk-
worm, Bombyx mori, with a fungal pathogen, later named
Beauveria bassiana (Bassi, 1835). Bassi is known as the
“Father of Insect Pathology” for demonstrating the trans-
mission of the fungus from larva to larva and from con-
taminated leaves to larvae (Steinhaus, 1975; Davidson,
2012). Subsequently, a number of entomopathogens have
been incriminated as the causal agents of insect disease,
including Louis Pasteur’s landmark demonstration of the
microsporidium Nosema bombycis as the causal agent of
pébrine disease in B. mori (Pasteur, 1870). According to
Steinhaus (1949), in solving this problem and another
entomopathogen-related silkworm disease (flacherie),
Pasteur saved the French silk industry. Metchnikoff
was the first to culture an entomopathogen, the fungus
Metarhizium anisopliae, on artificial medium after isola-
tion from the wheat cockchafer Anisoplia austriaca and
to suggest using it as an applied microbial control agent
(MCA) (Metchnikoff, 1879, 1880; Steinhaus, 1975; Lord,
2005). However, it was not until 1888 that M. anisopliae
Microbial Control of Insect and Mite Pests. http://dx.doi.org/10.1016/B978-0-12-803527-6.00001-9
Copyright © 2017 Elsevier Inc. All rights reserved.
was evaluated as an MCA (Krassilstschik, 1888). Since
then, a multitude of entomopathogens have been discov-
ered and developed as MCAs (Steinhaus, 1963, 1975;
Lord, 2005; Davidson, 2012; Lacey et al., 2015).
The first commercialization of MCAs began in France
in 1938 with Sporeine, a product based on the bacte-
rium Bacillus thuringiensis. The production and use of
the product were discontinued, ostensibly due to World
War II (Steinhaus, 1949; Davidson, 2012). The first ento-
mopathogenic bacterium to be registered in the United
States (1948) was Paenibacillus (Bacillus) popilliae (also
known as milky spore) for control of the Japanese beetle,
Popillia japonica (Dutky, 1963; Klein, 1981; Federici,
2005; Davidson, 2012). It was applied as an augmenta-
tive MCA to larval breeding sites of P. japonica larvae,
especially in residential lawns and golf courses. It is no
longer commercially produced due in large part to the
requirement of in vivo production. Steinhaus stimulated
renewed interest in the development of B. thuringiensis
and other MCAs with the publication of “Living Insecti-
cides” (Steinhaus, 1956). Although the first commercial B.
thuringiensis product was registered for use in the United
States in 1961, it was not until the discovery and com-
mercial development of efficacious strains with greater
virulence toward a broader range of pests that microbial
control became widely used in agriculture and forestry
(Kurstak, 1962; Dulmage, 1970; Glare and O’Callaghan,
2000). More detail on the development of this key MCA
leading to its widespread use is presented in Chapter 4
and by Federici (2005) and by Sanchis (2011). Historical
accounts of the development of other entomopathogens
as MCAs are presented by Steinhaus (1949, 1975), Lord
(2005), and Davidson (2012). Several significant advances
in microbial control of pest insects and mites have taken
place since the comprehensive reviews by Burges and
Hussey (1971) and Burges (1981). Glare et al. (2012) and
Lacey et al. (2015) reviewed the successes and setbacks
for microbial control and made recommendations for
improvements of MCAs and their future use.
3
4  PART | I  Introduction to Insect Pathology and Microbial Control: The Tried and True and Recent Innovations
1.2 ENTOMOPATHOGENS USED AS MCAs
1.2.1 Entomopathogenic Viruses
A wide array of viruses have been reported from a broad spec-
trum of insects and other arthropods (Miller, 1997; Miller and
Ball, 1998). These include representatives from at least 16
families of viruses (Possee and King, 2014) including bacu-
loviruses, cytoplasmic polyhedroviruses, entomopoxviruses,
nudiviruses, and a variety of lesser known viruses that have
been investigated for their potential as MCAs. The most
researched for microbial control are the baculoviruses with
over 600 species of insects being reported as hosts (Miller,
1997; Szewczy et al., 2006; Cory and Evans, 2007). Inunda-
tion augmentative application is the most commonly adopted
strategy for their use. Of the four genera of the Baculoviridae,
the Alphabaculovirus [nucleopolyhedroviruses (NPVs)] and
Betabaculovirus [granuloviruses (GVs)] are the most numer-
ous and most widely used as MCAs (Cory and Evans, 2007;
Eberle et al., 2012). NPVs are predominantly found in the
Lepidoptera, but also in Diptera, Hymenoptera (Symphyta),
and some Crustacea while GVs infect only Lepidoptera
(Miller, 1997; Eberle et al., 2012). In addition to host range,
the two genera differ mainly in the manner in which virions
are embedded in the infectious particles, the proteinaceous
occlusion bodies (OBs). In NPVs, several viral rods are
embedded together within the same large polyhedral OBs,
whereas in GVs only a single virion is embedded singly in
smaller OBs, sometimes called a granule. In NPVs, viral
rods can be singly enveloped (SNPV) or multiply envel-
oped (MNPV). Morphology of GVs, MNPVs, and SNPVs
is depicted in Chapter 3, Figure 3.2. MNPVs usually have
broader host ranges than SNPVs. For example, the Autogra-
pha californica MNPV (AucaMNPV) is active against larvae
of 95 species in 15 families of Lepidoptera (Vail et al., 1971,
1999; Vail and Jay, 1973; Cory and Evans, 2007). However,
it is most efficacious against species in the Noctuidae (Vail
et al., 1999). In contrast, another MNPV, Lymantria dispar, is
strictly host specific (Barber et al., 1993). Selected NPVs and
GVs are presented in Table 1.1, and a more extensive listing
is presented in Chapter 3.
With few exceptions, baculovirus OBs must be ingested
in order to invade the host insect. Virions enter the host via
the midgut epithelium where they initially multiply before
spreading and infecting other types of cells (fat body, tracheal
matrix, hypodermis) and begin viral replication and occlu-
sion. Detailed descriptions of the infection processes and
pathogenesis are presented by Federici (1997) and in Chapter 3.
The macroscopic progression of disease signs and symptoms
are shown in Chapter 3, Figure 3.1. Among all entomopatho-
genic viruses, baculoviruses are the only ones to have been
mass produced, commercialized, and applied on a widespread
basis. The issues and constraints around expanding their use on
a broader scale are presented in Chapter 3. Mass production of
baculoviruses is covered in Chapter 7.
1.2.2 Bacteria
Although the major volume and market share of commer-
cially produced MCAs that are used for microbial control
are bacteria, relatively few species have been developed.
These include gram-positive, spore-forming B. thuringien-
sis subspp. and L. (B.) sphaericus, the gram-negative, non–
spore-forming Serratia entomophila, and, most recently,
Chromobacterium subtsugae. B. thuringiensis subsp. are
the most commonly used of all microbial pesticides world-
wide, representing over 50% of all MCA sales (Glare et al.,
2012; Lacey et al., 2015). The majority of B. thuringiensis
inundative applications are for suppression of lepidopteran
pests of crops followed by applications for control of for-
est pests and then by abatement of mosquitoes and black
flies. The insecticidal activity of B. thuringiensis and
L. sphaericus is mainly due to protein toxins within para-
sporal inclusions, which must be ingested to become lar-
vicidal. The toxins become solubilized in the high pH of
the midgut and are cleaved to the toxic moieties by gut
proteolytic enzymes. These toxins recognize receptors and
create pores on enterocytes, with the subsequent osmotic
imbalance resulting in cell rupture, compromising integrity
of the gut epithelium. Invasion of the main body cavity and
subsequent septicemia caused by gut-resident bacteria leads
to death of the larva soon afterward. Detailed descriptions
of the biochemical nature of the toxins and their modes of
action at the molecular level are presented in Chapter 4.
Examples of targeted pests for inundative applications
include a plethora of lepidopteran pests of field crops, tree
fruit, turf, and ornamental plants and forest and dipteran
pests and are shown in Table 1.1. The larvicidal activity
S. entomophila is due to proliferation of the bacterium in the
gut resulting in blockage and lack of nutrient absorption.
More details on host range factors affecting larvicidal activ-
ity, safety, and production of entomopathogenic bacteria are
presented in Chapters 4 and 8.
1.2.3 Fungi
Depending on the scientists reporting, between 700 and
1000 species of fungi are recognized as causal agents of dis-
ease in arthropods (Goettel et al., 2010; Vega et al., 2012).
The majority of those used for microbial control of insects
and mites are in the orders Hypocreales and Entomophtho-
rales. In addition to a broad host range within the Insecta
and Acarina, they are the only effective MCAs of sucking
insects (Hemiptera). The only commercially produced fungi
for microbial control are in the Hypocreales. These include
Beauveria bassiana, Metarhizium spp., Isaria fumosorosea,
and Lecanicillium spp. and are used against dozens of pests
in a variety of crops.
The typical route of invasion of the host is through the
cuticle. Once fungal spores have germinated on the surface
of the host, the growing tip (appressorium, infection peg,
 Introduction to Microbial Control Chapter | 1  5

TABLE 1.1  Some Examples of Entomopathogens Used for Microbial Control of Targeted Arthropods

Pathogen Group Pathogens Targeted Arthropods Crops/Habitats

Viruses: Baculoviridae

Alphaviruses-NPV HearSNPV, HezeSNPV Helicoverpa and Heliothis spp. Corn, cotton, soybean,
tobacco
SeMNPV Spodoptera exigua Vegetable, field, flower crops,
and ornamentals

Betaviruses-GV CpGV Cydia pomonella Pome fruit and walnut

AdorGV Adoxophyes orana Apple

Bacteria

Bacillus thuringiensis subsp. Lepidoptera Vegetables, tree fruit, forest,

kurstaki and aizawai stored products

B. thuringiensis subsp. Diptera: Culicidae, Simuliidae Lotic and lentic aquatic

israelensis habitats

Lysinibacillus sphaericus Diptera: Culicidae Lentic aquatic habitats

Serratia entomophila Costelytra zealandica Pasture

Chromobacterium subtsugae Broad host range including: Diverse crops

Leptinotarsa decemlineata,
Hemiptera, Acarina
Fungi
Hypocreales Beauveria bassiana Broad host range including: Diverse crops: Cucurbits,
Lepidoptera, Coleoptera, potato, crucifers, cotton,
Formicidae, Hemiptera, Acarina sugarcane, and more

Metarhizium anisopliae senso Very broad host range including: Diverse crops/habitats and
lato Coleoptera, Diptera, Lepidoptera, structures
Hemiptera, Isoptera, Acarina

Metarhizium acridum Grasshoppers and locusts Diverse crops

Isaria fumosorosea Hemiptera Diverse crops including:

Cucurbits, cotton, crucifers,
and ornamentals

Lecanicillium spp. Hemiptera Diverse crops, especially

greenhouse crops
Entomophthorales

Neozygites tanajoae Mononychellus tanajoa Cassava

Entomophaga maimaiga Lymantria dispar Deciduous trees

Entomopathogenic Nematodesa

Steinernema carpocapsae Lepidoptera, Coleoptera, Diptera, Field crops, orchard,

Hymenoptera ornamental and turf

S. feltiae Coleoptera, Lepidoptera, Vegetables, tree fruit,

ornamentals, mushrooms

S. scapterisci Orthoptera (Scapteriscus spp.) Lawn and turf

Heterorhabditis bacteriophora Lepidoptera, Coleoptera Vegetables, orchard,

ornamentals, turf mushrooms

H. megidis Coleoptera (Scarabaeidae) Lawn and turf

aInsect orders in which naturally occurring EPN infections were found (Peters, 1996). Experimental host ranges can be narrow (H. megidis, S. scapterisci)

to very broad (S. carpocapsae, S. feltiae, H. bacteriophora). See also Chapter 6.

6  PART | I  Introduction to Insect Pathology and Microbial Control: The Tried and True and Recent Innovations
germ tube) penetrates the cuticle through concentrated physi-
cal energy and lytic enzymatic activity (Hajek and St. Leger,
1994). Following penetration and growth of hyphae and blas-
tospores within the host, sporulation of the fungus takes place
on the surface of the cuticle. Selected examples of the use
of entomopathogenic fungi for protection of crops and urban
structures are presented in Table 1.1 and in Chapter 5 and
several of the applied chapters in the book. Mass production
of entomopathogenic fungi is presented in Chapter 9.
1.2.4 Entomopathogenic Nematodes
A huge number of entomogenous nematodes have been
reported from insects, (Welch, 1963; Poinar, 1975; Grewal
et al., 2005) including those used in classic biological con-
trol (Bedding and Akhurst, 1974; Platzer, 2007; Bedding,
2009; Frank, 2009). The only nematodes currently used
as MCAs for augmentative biological microbial control
are entomopathogenic nematodes (EPNs) in the families
Steinernematidae and Heterorhaditidae. Species of EPNs
are in mutualistic and obligate associations with bacteria
(Xenorhabdis spp. and Photorhabdis spp.) carried inside the
gut of infective juveniles (IJs). These IJs wait for or actively
seek a host insect. They gain entry into the host insect, per
os, via the spiracles or in some cases by penetrating inter-
segmental membranes. Inside the host, the mutualistic bac-
teria are liberated, kill the host, and digest host tissues, and
the IJs molt and begin feeding. There are normally two to
three generations, although if nutrients are low, only one
generation may result. Once nutrients are exhausted, IJs are
produced and leave the host insect to search for or wait for
new hosts. Some examples of EPNs, insect targets, and pro-
tected crops are presented in Table 1.1. Detailed information
on the life cycle, host ranges, factors that enhance or retard
EPN efficacy, and protected crops are given in Chapter 6.
Mass production of EPNs is covered in Chapter 10.
1.3 MICROBIAL CONTROL AGENTS AND
BIOLOGICAL CONTROL
Entomopathogens used as MCAs fit the three categories of
biological control: classic, conservation, and augmentative,
as defined by Barbosa (1998), McCrevy (2008) and Hoy
(2008a,b) for natural enemies of pest arthropods. Entomo-
pathogens have been successfully used in all three categories.
Classic biological control entails introduction of naturally
occurring predators, parasitoids, and entomopathogens from
the native range of an invasive pest to help bring about its long-
term suppression; conservation biological control stresses
maintaining a habitat that is conducive to conserving natu-
ral enemies of the pest; and augmentative biological control
involves enhancing the prevalence of the natural enemies to
initiate increased control. In microbial control, this is accom-
plished with either inoculative or inundative application of
MCAs. Eilenberg et al. (2001) proposed replacing augmenta-
tive biological control with two separate categories: inocula-
tive biological control and inundative biological control.
1.3.1 Classic Biological Control
There are numerous examples of MCAs used for classic bio-
logical control of invasive insects (Hajek et al., 2005, 2007,
2009). These include entomopathogenic viruses (baculovi-
rus, nudivirus), fungi (Entomophthorales), and parasitic and
entomopathogenic nematodes (Neotylenchidae, Mermithi-
dae, Steinernematidae). Three noteworthy examples of clas-
sical microbial control follow.
1.3.1.1 Case Study: Oryctes Nudivirus for
Control of Invasive Coconut Palm
Rhinoceros Beetle
One of the most impressive examples of classic biological
control with an MCA is that of the Oryctes nudivirus for
control of the coconut palm rhinoceros beetle, Oryctes rhi-
noceros (Hüger, 2005; Jackson et al., 2005; Jackson, 2009).
The beetle is a serious exotic pest of coconut and oil palms
in the southwest Pacific islands. Adults consume foliage
in the crown of the palms and can reduce yield and kill
trees (Bedford, 1980). Larvae develop in rotting palm logs,
including the tops of standing dead palms, and in organic
matter in a diversity of other sites with high organic con-
tent (sawdust, manure piles, etc.) (Bedford, 1980). Native to
the coconut growing areas of the Asia–west Pacific region,
the beetle was accidentally introduced into Samoa and sub-
sequently spread to other islands in the southwest Pacific,
where it became a widespread and very serious pest of
coconut palms (Bedford, 1980; Jackson, 2009).
The Oryctes virus was originally collected from infected
Oryctes rhinoceros in Malaysia by Hüger (1966) and intro-
duced into Samoa and several southwest Pacific islands
(Bedford, 1980; Hüger, 2005; Jackson, 2009). Adult beetles
become chronically infected and serve as mobile reservoirs
of the virus. Aggregations of mating and feeding adults
in the palm crowns ensures transmission from infected to
uninfected individuals and dispersal of the virus. Larval
breeding sites become initially contaminated with virus
via ovipositing infected females. Larvae become acutely
infected after consuming virus and die within 9–25 days
depending on age and temperature (25–32°C) (Hüger, 1966;
Zelazny, 1972). The introduced virus has resulted in signifi-
cant long-term control of the beetle and reduction of foliar
damage by causing epizootics that kill larvae, curtail the life
span of adults, and reduce fecundity in females (Zelazny,
1972, 1973; Bedford, 1980; Hüger, 2005; Jackson, 2009).
Production and application of the virus is reviewed by
Bedford (1980) and Jackson (2009). In recent years there
are reports of Oryctes virus having become attenuated and

therefore less effective on some of the beetle-infested islands
(Jackson et al., 2005; Jackson, 2009). Jackson et al. (2005)
and Jackson (2009) conclude that selection of more virulent
strains of virus and improvements in application methods
need to be developed to overcome this problem.
1.3.1.2 Case Study: Entomophaga maimaiga
for Classic Biological Control of Gypsy
Moth, L. dispar
An example of classic biological control using a fungus as
an effective pathogen for long-term control of the gypsy
moth, L. dispar, is that of the entomophthoralean E. mai-
maiga (Hajek et al., 1996; Hajek, 1997; Solter and Hajek,
2009). The moth is a native forest defoliator that outbreaks
occasionally from Europe to Asia but was absent from the
Western Hemisphere. From its original point of entry into
the United States (Boston, Massachusetts) in 1869, it has
dispersed annually farther south and west. Larvae of L.
dispar feed on a wide range of deciduous trees and during
cyclic outbreaks have been responsible for defoliation of
trees in up to 2 million ha of forest (Solter and Hajek, 2009).
The fungus originated in Japan and was first released
in the United States near Boston in 1910 and 1911 but was
not detected in following years (Hajek et al., 1995). It was
also collected in Japan in 1984 by Soper et al. (1988), and
small field applications of the 1984 isolate in 1985 in New
York State and in 1986 in Virginia resulted in few or no
infections. It was not until 1989 that significant epizoot-
ics caused by E. maimaiga were reported (Andreadis and
Weseloh, 1990; Hajek, 1997, 1999). The exact origin of
the E. maimaiga causing these outbreaks is not precisely
known, but it appears to have originated from areas in Japan
different from the 1910–1911 collection sites (Nielsen
et al., 2005). Since then, in vivo produced fungal spores
(conidia and resting spores) and infected larvae have been
introduced into areas on the leading edge of L. dispar dis-
persal. Subsequent infections indicate that E. maimaiga has
become established in and spread from locations where it
was not previously observed (Hajek et al., 1996; Weseloh,
2003; Solter and Hajek, 2009). More details on this con-
tinuing successful classic microbial control are presented
in Chapter 21.
1.3.1.3 Case Study: Steinernema scapterisci for
Control of Invasive Mole Crickets
Mole crickets, Scapteriscus spp., expanding northward
from their centers of origin in South America, were intro-
duced into the United States in the early 1900s (Frank,
2009). Since introduction, severe damage had been
reported in turf infested by the crickets, especially in
Florida, where S. vicinus is a serious pest of turf (Frank,
2009). Exploration for EPNs and other natural enemies of
Introduction to Microbial Control Chapter | 1  7
Scapteriscus spp. were started in South America. In 1985,
an EPN from Uruguay was released in Florida. Initially,
the introduction of the EPN, S. scapterisci, for control of
the invasive mole crickets in Florida fit the classic biologi-
cal control paradigm (Parkman et al., 1993). The nematode
was collected in Uruguay and introduced into S. vicinus, S.
borelli, and S. abbreviatus populations in Florida where it
became established (Hudson et al., 1988; Parkman et al.,
1993). Additionally, two imported parasitoids, also from
South America, were released and became established
throughout Florida. By 2000, the combined effort of the
three natural enemies reduced the Scapteriscus populations
by 95% (Frank and Walker, 2006). Since then the EPN has
been applied to infestations of the cricket in several sites in
Florida (Frank, 2009).
Numerous additional examples of classic microbial
control are found throughout the literature; some of which
are examined in this book. These include importation and
establishment of the nematode Deladenus siricidicola
(Tylenchida: Neotylenchidae) for control of the Sirex wood-
wasp, Sirex noctilio (Chapter 21); importation and establish-
ment the of nematode Romanomermis iyengari (Nematoda:
Mermithidae) for control of mosquitoes (Chapter 28); devel-
opment of Neozygites tanajoae (Entomophthorales) for
control of cassava greenmite in Brazil and Chapter 2 Chap-
ter 21Benin (Chapter 2); baculoviruses in forestry (Chap-
ter 21); and agriculture (Chapters 14 and 17). A catalog of
imported exotic entomopathogens used as classic biological
control agents was compiled by Hajek et al. (2005).
1.3.2 Conservation Biological Control
Epizootics caused by naturally occurring viral and fungal
pathogens are often responsible for spectacular crashes of
insect pest populations and are often credited with elimi-
nating the need for further interventions (Harper, 1987;
Steinkraus, 2007; Shapiro-Ilan et al., 2012). Conservation
biological control relies on the protection of and improving
conditions for naturally occurring MCAs to enable induc-
tion of high rates of infection in pest populations. Reliance
on the natural occurrence of entomopathogens for pest man-
agement, however, can be risky due to the unpredictability
of factors that govern epizootics. Because many pathogens
are host-density dependent, epizootics often occur after
economic injury levels have been surpassed (Odindo, 1983;
Harper, 1987; Rose et al., 2000). However, agricultural
practices that foster their conservation and increase their
prevalence should nevertheless be considered.
A demonstration of the conservation microbial con-
trol strategy is that reported by Steinkraus (2007). By not
treating cotton with broad-spectrum insecticides, popula-
tions of the cotton aphid, Aphis gossypii, were allowed
to develop, which in turn encouraged epizootics of the
fungus Neozygites fresenii. Conservation of naturally
8  PART | I  Introduction to Insect Pathology and Microbial Control: The Tried and True and Recent Innovations
occurring MCAs can also be disrupted by the effect of
pesticides directly on the MCAs. For example, application
of fungicides and nematicides can drastically curtail the
insecticidal activity of entomopathogenic fungi and EPNs,
respectively.
In addition to pesticide type and timing of application,
other agricultural practices [type of tillage (conventional
versus conservation), irrigation, fertilizer (chemical ver-
sus organic) crop rotation, type of ground cover, etc.] can
have a marked effect on the survival and pathogenicity of
MCAs. Hummel et al. (2002) and Millar and Barbercheck
(2002) found that tillage practices can significantly affect
the survival and abundance of entomopathogenic fungi and
EPNs. Hummel et al. (2002) demonstrated that detection of
entomopathogens was significantly higher in conservation
tilling compared with conventional tillage systems. They
reported that conservation tilling (strip-till) did not affect
levels of detection of Steinernema carpocapsae, but pes-
ticide use significantly reduced detection of entomopatho-
genic fungi. Millar and Barbercheck (2002) demonstrated
that tillage versus no tillage resulted in a significant negative
effect of detection of S. carpocapsae but a positive effect on
detection of S. riobrave. The authors surmised that the dif-
ferent sensitivities of the EPNs could be partly explained by
differences in environmental tolerances of the two species
and tendencies to disperse deeper into the soil. A number
of other examples of the role of conserved EPN and fungal
MCAs in microbial control are presented by Lewis et al.
(1998), Ekesi et al. (2005), Meyling and Eilenberg (2007),
Nielsen et al. (2007), Steinkraus (2007), Pell et al. (2010),
and Campos-Herrera et al. (2010).
1.3.3 Augmentative Biological Control
Augmentation of naturally occurring MCAs can be accom-
plished with either inoculative or inundative applications
depending on the pathogen and crop that requires protec-
tion. Inoculative microbial control is the application of
smaller amounts of inoculum with the goal that the MCA
will increase in the host population on its own (Eilenberg
et al., 2001). This could result in outbreaks of disease ear-
lier in the season than would otherwise occur. Examples
include limited applications of P. popilliae and Heterorhab-
ditis bacteriophora for control of Popillia japonica (Dutky,
1963; Klein, 1981; Klein and Georgis, 1992).
The more common strategy is the inundative applica-
tion of larger amounts of MCAs to initiate widespread
infections and provide immediate control of targeted pests
(Steinhaus, 1949, 1963; Burges and Hussey, 1971; Burges,
1981; Lacey et al., 2001, 2015; Kaya and Lacey, 2007).
In the inundative approach, the main route of infection is
through the pests picking up the applied MCA rather than
through secondary infections. Depending on the number
of generations of the pest insect or mite and the impact of
adverse environmental factors on MCA persistence, espe-
cially exposure to ultraviolet-B radiation, one to several
applications of the MCA may be required throughout the
growing season.
There are scores of commercially produced microbial
pesticides used for augmentative control of pest insects
and mites worldwide (Glare and O’Callaghan, 2000; Fed-
erici, 2005; Alves and Lopes, 2008; Kabaluk and Gazdik,
2005; Faria and Wraight, 2007; Gwynn, 2014; Lacey et al.,
2015). Some examples of the augmentative use of ento-
mopathogenic viruses, bacteria, fungi, and nematodes are
presented in Table 1.1. B. thuringiensis is the most widely
used MCA for control of hundreds of species of insect pests
(Glare and O’Callaghan, 2000; Federici, 2005; see Chapter 4).
The use of MCAs against insects and mites is the predomi-
nant theme of Chapters 11–29.
1.4 ADVANTAGES AND DISADVANTAGES
OF MICROBIAL CONTROL
Table 1.2 presents several advantages and disadvantages
of microbial control as compiled from Steinhaus (1949),
Burges and Hussey (1971), Tanada and Kaya (1993),
Alves (1998), Kaya and Lacey (2007) and Kaya and Vega
(2012). The most important advantages are the efficacy,
specificity, and safety of MCAs. In contrast, the greatest
disadvantages include the higher comparative cost, nar-
rower spectrum of insecticidal activity, and reduced per-
sistence relative to conventional chemical insecticides.
However, benefits of environmental and food safety are
increasingly having a positive effect on the sales and
popularity of microbial control. Numerous studies attest
to the safety of MCAs for applicators and other verte-
brates, most nontarget organisms, especially honeybees
and predators and parasitoids (Hokkanen and Hajek,
2003). There could be indirect effects on other natural
enemies if the host insect is killed before parasitoids
emerge or host and prey are significantly removed from
the food chain. However, natural enemy vagility and
alternative hosts and prey minimize this effect so that the
overall positive effect of arthropod natural enemies on
pests outweighs any potential negative effects. Publica-
tion of studies on the safety of MCAs have been reviewed
by Laird et al. (1990), Glare and O’Callaghan (2000),
Lacey and Siegel (2000), Goettel et al. (2001), and Hok-
kanen and Hajek (2003). MCAs may not always provide
a stand-alone means for total pest control, yet they can
be invaluable components of integrated pest management
working in concert with other natural enemies, resistant
plant varieties, agricultural practices, the judicious use of
selective and soft pesticides (avermectin, spinosad, etc.),
mating disruption, and environmental manipulation and
modification (Gurr et al., 2004; Radcliffe et al., 2009;
Wraight and Hajek, 2009).
 Introduction to Microbial Control Chapter | 1  9

TABLE 1.2  Advantages and Disadvantages of Using Microbial Control Agents (MCAs)

Advantages
Specificity, minimal effects on nontarget insects
Efficacious for targeted insects and mites
No development of resistance with most MCAs
Recycling and long-term persistence of some pathogens enabling
long-term control
No maximum residue limits (MRL) for MCAs on produce
Little or no secondary pest outbreaks due to preservation of
natural enemies
Effective tool for resistance management in IPM strategies
Safe for applicators, other vertebrates and the food supply
No preharvest spray interval
Little or no environmental pollution
Ease of mass production of nonobligate pathogens on inexpensive
artificial media
Good shelf-life of most formulated MCAs
Application with conventional equipment
Adaptable to genetic modification
Disadvantages
Specificity, less effective on pest complexes
Short shelf-life for some unformulated MCAs
Sensitive to inactivation by ultraviolet and other environmental
conditions.
Multiple applications may be required due to short persistence of
MCA on crops
May require precise timing of applications to infect early instars
Slower onset of death relative to chemical pesticides
Development of resistance of some Lepidoptera to B. thuringiensis
and of Cydia pomonella to the codling moth granulovirus
In vivo production costs could be high
Uneconomical except for high value crops
Potential for negative public perception of entomopathogens used
on crops
Negative public reaction to the use of genetically modified MCAs

Information compiled from Steinhaus, E., 1949. Insect Pathology, Academic Press, 757 pp.; Burges, H.D., Hussey, N.W., 1971. Microbial Control of Insects
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2007. Introduction to microbial control. In: Lacey, L.A., Kaya, H.K. (Eds.), Field Manual of Techniques in Invertebrate Pathology: Application and Evaluation
of Pathogens for Control of Insects and Other Invertebrate Pests, second ed. Springer, Dordrecht, The Netherlands, pp. 3–7; Kaya, H.K., Vega, F.E., 2012.
Scope and basic principles of insect pathology. In: Vega, F.E., Kaya, H.K. (Eds.), Insect Pathology, second ed. Academic Press, San Diego, pp. 1–12.

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