Biocontrol Science and Technology

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Prospective Biological Control

Agents of Varroa destructor
n. sp., an Important Pest of
the European Honeybee, Apis
mellifera
a a b
D. Chandler , K. D. Sunderland , B. V. Ball & G.
a
Davidson
a
Department of Entomological Sciences,
Horticulture Research International, Wellesbourne,
Warwick, CV35 9EF, UK
b
Department of Entomology and Nematology, IACR
Rothamsted, Harpenden, Herts, AL5 2JQ, UK
Published online: 28 Jun 2010.
To cite this article: D. Chandler , K. D. Sunderland , B. V. Ball & G. Davidson (2001):

Prospective Biological Control Agents of Varroa destructor n. sp., an Important Pest
of the European Honeybee, Apis mellifera, Biocontrol Science and Technology, 11:4,
429-448
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Biocontrol Science and Technology (2001) 11, 429 ± 448
REVIEW
Prospective Biological Control Agents of Varroa destructor

n. sp., an Important Pest of the European Honeybee,
Apis mellifera
D. CHANDLER,1 K. D. SUNDERLAND,1 B. V. BALL2 and

G. DAVIDSON1
1
Department of Entomological Sciences, Horticulture Research International,
Wellesbourne, Warwick, CV35 9EF, UK; 2Department of Entomology and
Nematology, IACR Rothamsted, Harpenden, Herts AL5 2JQ, UK
(Received for publication 9 September 2000; revised manuscript accepted 26 January 2001)
This paper reviews prospective biologica l control agents of the varroa mite, Varroa destructor
n. sp. (Acari, Mesostigmata). This ectoparasite has caused severe damage to population s of
the European honeybee, Apis mellifera, world-wide in recent years. To date, no promising
natural enemies of varroa species have been identiWed on A. mellifera or its original host,
Apis cerana. Therefore, biologica l control will probably require natural enemies from other
hosts. The following groups of organisms were reviewed as potential biologica l control agents:
predatory mites, parasitoid s and entomopathogen s (nematodes, protozoa, viruses, Bacillus
thuringiensis, rickettsiae, and fungi). The candidate groups were ranked according to their
lethality to Acari, likely ability to operate under the physical conditions of honeybee colonies,
ease of targeting, and ease of mass-production . Preferential consideration was given to the
natural enemies of Acari that occupy taxonomic groups close to varroa. Entomopathogeni c
fungi, which kill a wide range of acarine species, were identiWed as prime candidates for
screening against varroa. Bacillus thuringiens is also requires study, particularl y strains
producing novel toxins active against non-insect hosts. Entomopathogeni c protozoa and nemat-
odes show less potential for varroa control, but nonetheless warrant preliminary investigation.
We consider predators, parasitoids, viruses and rickettsiae to have little potential to control
varroa. Because the physical conditions within honeybee colonies are similar everywhere, it
is very likely that a biological control agent of varroa could be used successfully throughou t
the world.
Keywords: Varroa destructor, Varroa jacobsoni, Apis mellifera, Acari, biologica l control,
entomopathogens, nematodes, protozoa, viruses, Bacillus thuringiensis, rickettsiae, fungi,
predators, parasitoid s
Correspondence to: D. Chandler. E-mail: david.chandler@hri.ac.uk
ISSN 0958-3157 (print)/ISSN 1360-047 8 (online)/01/040429-20 2001 Taylor & Francis Ltd
DOI: 10.1080 /0958315012006747 2
430 D. CHANDLER ET AL.
INTRODUCTION
The varroa mite, Varroa destructor n. sp. (Acari, Mesostigmata) is a damaging parasite of
the European honeybee, Apis mellifera (Anderson & Trueman, 2000). This pest originates
in Asia, where it is a parasite of the Eastern honeybee, Apis cerana, but it has extended its
host range to A. mellifera and has caused severe damage to population s of this species
world-wide in recent years (Ball, 1994a; Oldroyd, 1999). It entered Europe in the 1970s, the
USA in 1987 and the UK in 1992 (De Jong et al., 1982; Del® nado-Bake r & Houck, 1989;
Walton, 1996; Martin, 1997). At present, beekeepers attempt to control varroa with chemical
pesticides, but resistance to these chemicals is developing (Thomas, 1997; Elzen et al., 1998)
and alternative, sustainable methods of control are required urgently.
Until recently, V. destructor n. sp. was called Varroa jacobsoni Oudemans. However,
V. jacobsoni is now considered to be a complex of at least two species infesting A. cerana,
and the new species name has been proposed (Anderson & Trueman, 2000). Varroa jacobsoni
and V. destructor n. sp have been separated on the basis of morphology, reproductive

isolation , and mtDNA gene sequences (Anderson & Trueman, 2000). Both species are
parasites of A. cerana but V. destructor n. sp. is the species also causing damage to
A. mellifera world-wide. The type specimen of V. jacobsoni was collected in Java and does
not parasitize A. mellifera (Oldroyd, 1999). Two haplotypes of V. destructor n. sp. originating
from mainland Asia are associated with A. mellifera (Anderson & Trueman, 2000). The
most common haplotype, referred to as the Korean haplotype, has been identi® ed on
A. mellifera in Europe (including the UK), Africa, Asia (including the Middle East) and the
Americas. A less common haplotyp e has been identi® ed on A. mellifera in Japan, Thailand,
and the Americas. Note however that the past scienti® c literature refers only to V. jacobsoni,
although papers concerning its interactions with A. mellifera (which form the bulk of the
varroa research covered in this review) apply to V. destructor n. sp. (Anderson & Trueman,
2000).
Varroa mites feed on honeybee haemolymph, and transmit diseases that reduce bee
longevity, lower reproductive capacity, and induce deformities (Ball, 1993, 1994a,b ; Martin,
1997; Bowen-Walker et al., 1999). The lifecycle is haplodiploid , and reproduction occurs in
the brood cells of the colony. Details of the lifecycle are given by De Jong et al. (1982),
Ramirez (1987), Beetsma (1994), Donze and Guerin (1994) and Sammataro et al. (2000).
Mated female mites enter brood cells to oviposit shortly before the cells are capped. The
® rst egg to be laid is unfertilized and develops into a neotenic male that remains in the cell.
Subsequent eggs are fertilized and develop as females, which mate with their brother soon
after maturation . The developin g mites feed from the haemolymph of the bee larva/pupa
through a single hole constructed and tended by the parent mite. Adult females emerge with
the young bee 12± 14 days after capping. On A. cerana, the reproduction of V. jacobsoni is
restricted to drone brood and does little harm. Population s of V. destructor n. sp. that aVect
A. mellifera, however, also reproduce on worker brood, which is extremely debilitatin g to
the colony (Oldroyd, 1999). Reductions in the numbers of worker bees leads to poor brood
care, reduced colony homeostasis, and a concomitant increase in brood diseases (Ball,
1994b).
The ecological and economic impact of varroa is considerabl e. The winter kill of managed
honeybee colonies by varroa was estimated at 13 million colonies world-wide in 1996
(Sanford, 1996), equivalen t to a quarter of the global commercial population . Colony losses
of up to 65% have been reported in some countries (Matheson, 1994). Apis mellifera is
utilized widely for crop pollinatio n and losses due to varroa can impact signi® cantly on
agriculture and horticulture. Honeybee pollinatio n has been valued at ca. £7 billion ($11
billion) per annum to UK farming, for example (Walton, 1996) and at £12.5 billion ($19
billion) per annum in the USA (Beetsma, 1994). Loss of honeybees also aVects the pollinatio n
of wild plants. In the USA, for example, varroa has destroyed nearly all stocks of feral bees
(Martin, 1997) and is considered a threat to biodiversit y (Allen-Wardell et al., 1998).
BIOCONTROL OF VARROA 431
At present, varroa is controlled with pesticides, but resistance to these chemicals has
started to develop (Milani, 1994, 1999; Hillesheim et al., 1996; Thomas, 1997; Elzen et al.,
1998). The USA (with the exception of Florida) and UK currently permit only pyrethroids
for varroa control (Bew, 1992; Ball, 1994a; Sanford, 1999) which could increase the selection
pressure for resistance in these countries. Pyrethroids and organophosphoru s pesticides,
sprayed against varroa, have a propensity to accumulate in beeswax (Fries, 1997). Alternative
agents, such as organic acids and essential oils, and cultural control methods (heating, drone
trapping), are only partially eVective and are labour intensive (Fries, 1993; Mobus & De
Bruyn, 1993; Ritter, 1993; Ball,1994a ; Beetsma, 1994; Engels, 1994). The breeding of varroa-
tolerant bees is a longer term option (Beetsma, 1994; Buchler, 1994; Danka et al., 1995;
Harbo & Hoopingarne r, 1997). There is a risk, however, that any varroa tolerance bred into
commercial bee stocks could be dissipate d by interbreeding with feral bees.
BIOLOGICAL CONTROL
The biological control of varroa warrants attention as a partial or total alternative to
chemicals, and it could form part of an integrated pest management system (Nasr & Kevan,
1999). The use of pathogens to control varroa was proposed by Dietz and Hermann (1988)
but very little research had been done on biological control. Varroa appears to be relatively
free of natural enemies. Virus-like particles have been identi® ed in varroa which are
associated with reduced mite longevity but with no apparent eVect on A. mellifera hosts
(Kleespies et al., 2000). However, no natural enemies causing wholescale populatio n declines
of varroa have been identi® ed. Indeed, it is possible that honeybee colonies act as a refuge
for varroa from its potential natural enemies. Only small population s of varroa develop on
A. cerana (Oldroyd, 1999), hence varroa natural enemies are likely to be rare on this bee.
Large, densely-packe d population s of varroa develop on A. mellifera, and therefore oVer a
tangible resource for natural enemies, but it is unlikely that any new association has had
suYcient time to evolve. Varroa may also have escaped its parasites during host shifts to
A. mellifera. Whatever the mechanism, the shortage of existing natural enemies of varroa
means that biologica l control will require natural enemies from other hosts. This paper
assesses prospective biological control agents of varroa. The main criteria used to rank
candidate agents were: (i) lethality to Acari, especially against mites related phylogenetically
to varroa or which occupy a comparable trophic niche; and (ii) likely ability to operate
under the physical conditions of the colony. Ease of targeting against varroa and mass-
production are also considered important.
We have used Evans (1992) classi® cation of the Acari when considering the host range of
candidate agents. Varroa jacobsoni (Oudemans) and V. destructor n. sp. (Anactinotrichida ,
Mesostigmata, Varroidae ) are closely related to the Laelapidae (containing free-living mites
and bird/mammal ectoparasites) and the Iphiopsidae (mites associated with arthropods)
(G.O. Evans, pers. comm.). The Mesostigmata are an ecologically diverse group of mites
that includes free living and phoretic species (Evans, 1992), and we consider them to be key
hosts in our review. The Ixodida (ticks) and Holothyrida (tropical and subtropical predators)
are sister orders to the Mesostigmata (Evans, 1992) and we have also given them preferential
consideration. These three orders are described collectively as the Parasitiformes.
The physical environment of the honeybee colony will be a major determinant of the
success of any prospective biologica l control agent of varroa. Conditions within colonies are
regulated by bees and diVer signi® cantly in winter and summer. In summer, temperatures
within colonies are largely independent of ambient temperatures (Liu et al., 1990). The
brood area is usually maintained at 32± 37ë C (Ribbands, 1953; Free & Spencer-Booth, 1959;
Jay, 1964; Powell, 1979). The temperature in broodless parts of the colony varies from
28± 33ë C (Kaya et al., 1982). Summer bees appear to have wide limits of humidity tolerance
(Ribbands, 1953). Humidity is lowest in the brood area at 40± 50% relative humidity (RH)
but increases to 70% RH during bouts of evaporative cooling, and up to 100% RH when
the only aperture to the colony is a single bottom entrance (Simpson, 1961). Humidity is
likely to be higher in sealed cells than other areas of the colony, but this has not yet been
measured (Ramirez, 1987). Experimenters attempting to rear honeybee larvae in the
laborator y use conditions of 75± 100% RH (Jay, 1964).
Honeybees remain active within the colony in the winter to generate heat, and cluster
together to reduce the rate of heat loss (Free & Spencer-Booth, 1959; Simpson, 1961;
Winston, 1987). In temperate regions, the temperature within a winter cluster is normally
maintained at 20± 30ë C (Free & Spencer-Booth, 1959; Simpson, 1961; Butler, 1974), with a
daily ¯ uctuation of about 5ë C (Ribbands, 1953), but the temperature may drop occasionally
to ca. 13ë C (Simpson, 1961; Winston, 1987). The outside temperature of the cluster is
usually maintaine d at ca. 10ë C (Seeley, 1985), but it can fall to 4ë C (Dietz et al., 1988). The
humidity of winter clusters is 50± 85% RH, and is greater at the centre than at the periphery,
while below a winter cluster it is ca. 75% RH (Anderson, 1948). Fanning reduces humidity
¯ uctuations in the centre of the winter cluster (Simpson, 1961). In colonies with poor
ventilation, water can condense onto the inside of the hive walls (Butler, 1974).
Colonies maintain the carbon dioxide level at 0.1 to 4.3% during the summer (Winston,
1987) and at ca. 6% in winter (Simpson, 1961). The level inside sealed cells has not yet been
measured (Ramirez, 1987). Signi® cant movement of air within the colony could help to
distribute infective stages of varroa pathogens. Air¯ ows generated by convection within
colonies are only about 0.2 l s - 1 (Simpson, 1961), but fanning (in response to high
temperature or humidity) generates air¯ ows of 0.4± 1.0 l s - 1 through the colony (Simpson,
1961; Seeley, 1985).
The high temperatures within colonies in summer could be unfavourabl e for many
biologica l control agents, however some will be eVective under these conditions and there
are also opportunitie s to control varroa at other times of year. The best time for remedial
treatment varies on a regional basis (Delaplane, 1997). Delaplane and Hood (1999), for
example, identi® ed the best times for treatment with pesticides as late season (August) and,
to a lesser extent, early season (February) for varroa mites in the southeastern USA, based
on an analysis of economic thresholds. In other areas of the USA, treatments in spring and
autumn are recommended (Delaplane, 1997). DiVerent biological control agents of varroa
could be used at diVerent times of year, depending on their temperature tolerances.
SURVEY OF POTENTIAL BIOLOGICAL CONTROL AGENTS OF VARROA

Predatory Mites and Parasitoids
Predatory mites and parasitoids are used for the biologica l control of a range of insect and
acarine pests (Copping, 1998). They can be mass reared and give prolonged pest control
under favourable conditions. However, there are likely to be considerabl e physical, environ-
mental and biological constraints to the activities of any predator or parasitoid released in
honeybee colonies, and we do not consider them good candidates for varroa control.
Predatory mites that feed on Acari are located in 11 families of the Mesostigmata and
Prostigmata (Gerson & Smiley, 1990). They occupy a range of habitats, including honeybee
colonies (De Jong et al., 1982; Eickwort, 1988; Chmielewski, 1989, 1992). However, predatory
mites rarely feed on Mesostigmata in nature and we have been unable to identify any
specialist acarine predators of ectoparasitic mites or ticks from the literature. Generalist
acarine predators are unlikely to search bee bodies for adult varroa mites. They are unlikely
also to impact signi® cantly on varroa eggs or larvae, because the eVective period for entry
into the brood cell is short (ca. 20 h) (Fries et al., 1994) and conditions within the cell (high
temperatures and humidity, damage by bee larva, possibility of drowning in brood food)
may well be inimical to survival and predation. To consume varroa eggs or larvae, a predator
would have to enter a brood cell in tandem with an adult varroa mite, as entering earlier is
likely to lead to removal by worker bees on routine maintenance duties. There is the
signi® cant risk that any generalist predator capable of entering the brood cell would consume
bee eggs. Neoseiulus fallacis [Amblyseius fallacis] (Mesostigmata) has been recorded as prey
for the generalist predator Balaustium putmani (Prostigmata) in con® ned environments
(Laing & Knop, 1983), but this species inhabit s outdoor-temperate habitats (Gerson &
Smiley, 1990) and it is not known whether it would tolerate the high temperatures of a
honeybee colony in summer. Some generalist predatory mites can prosper in alien environ-
ments, although they have not been tested in honeybee colonies. For example, Pyemotes
tritici, a prostigmatid predator of stored product pests, controlled population s of the red
imported ® re ant, Solenopsis invicta, when introduced into its nests, and also predated
population s of the colonial caterpillar Malacosom a americanum in its webs (Bruce, 1983).
Some pyemotids attack bees and man, although others are considered safe for use (Bruce,
1983).
Eight species of hymenopteran parasitoids have been recorded from Acari. Of these,
Globulencyrtu s politus parasitizes actinotrichid mites, while the remainder are parasitoids of
ixodid ticks (Coineau & Davis, 1999). Ixodiphagu s hookeri [Hunterellus hookeri] is a
parasitoid of a number of ticks of medical and veterinary importance. A French strain of

this species was established in coastal Massachusetts in 1926, where it causes up to 26%
nymphal parasitism in black legged ticks, Ixodes scapularis (a blood-feeding ectoparasite of
mammals) (Lyon et al., 1998). In ® eld experiments in Kenya, I. hookeri controlled popula-
tions of Amblyomma variegatum feeding on cattle but did not control Rhipicephalus
appendiculatus (Mwangi et al., 1997). Given that I. hookeri exhibits host specialism within
the Ixodida, it is perhaps unlikely to parasitize mesostigmatid mites, including varroa.
Moreover, because the parasitoid reproduces within tick larvae and nymphs, it is unlikely to
have an eVect against varroa because access to the larvae is blocked by the brood cell cap.
Pathogens
Pathogens contribute to the natural regulation of many population s of arthropods. Much
of the research in this area concerns the causal agents of insect diseases and their exploitation
for biological pest control. Many entomopathogen s can be mass produced, formulated, and
applied to pest population s in a manner analogou s to chemical pesticides, i.e. as nonpersistent
remedial treatments that are released inundativel y. Entomopathogen s have also been used
as classical biological control agents of insect pests, and natural pest control by entomopatho-
gens has been enhanced by habitat manipulatio n (Fuxa, 1987; Hajek & St. Leger, 1994).
Comparatively little research has been done on the pathogens of Acari. These occur in the
same phylogenetic assemblages as entomopathogen s but are less diverse, which probably
re¯ ects host diversity (Poinar & Poinar, 1998). For the purposes of this review, we have
considered the pathogens of Acari to be a functional subgroup of the entomopathogen s
(Chandler et al., 2000).
While varroa has no reported existing lethal natural enemies, it may well be susceptible
to entomopathogens. A range of Acari from the Mesostigmata and related taxa are
susceptible to entomopathogen s (see below). It is noteworthy too that some insect pests are
rarely infected by pathogens in nature but are susceptible to microbial pesticides. For
example, cockroaches and termites can be controlled by applicatio n of the fungus Metarhiz-
ium anisoplia e but are rarely infected naturally by this entomopathoge n (Miller, 1990; Milner
& Staples, 1996). Similarly, larvae of the wax moth, Galleria mellonella, which inhabit
honeybee colonies and feed on comb, rarely encounter entomopathogen s naturally in
honeybee colonies but nonetheless are highly susceptible to them (Bedding & Akhurst, 1975;
Zimmerman, 1986).
Mesostigmatid mites can be infected by viruses, protozoa, fungi, nematodes and bacteria
(Steiner, 1993; Steiner & Bjornson, 1996). The Ixodida are also susceptible to pathogens,
particularly fungi (Chandler et al., 2000). There is, therefore, reasonable justi® cation to
search for microbial control agents of varroa. It should be noted, however, that honeybee
societies have evolved a number of ways of promoting nest hygiene, which may impair
pathogen performance. The high temperatures of honeybee colonies in summer may be
detrimental to many strains of microorganisms, while bee products, such as propolis, brood
food, and honey, have antibiotic properties which may aVect pathogens (Seeley, 1985;
Winston, 1987). Despite these precautions, bees suVer from diseases caused by fungi,
bacteria, protozoa and viruses (Ball, 1993, 1994a) . Since nest hygiene is fallible, therefore,
and is directed primarily agains t pathogens attacking bees and bee products, it would seem
likely that anti-varroa pathogens could be deploye d successfully in the honeybee colony,
given an appropriat e formulation and targeting strategy. The activities of many pathogens
are dose-dependent , and varroa mites form discrete, high density population s which should
be easy to target. Hence it may well be economically viable to use a weak pathogen at a
high dose, if no highly virulent pathogens can be identi® ed. Pathogens with contact activity,
such as fungi, are ® rst-choice candidate s for varroa control. Infections by orally-transmitte d
pathogens will have to be acquired from the surface of the host bee as the varroa
mite prepares to feed, and subsequently during feeding, which may be problematic. The
opportunitie s for acquiring orally-transmitte d pathogens could be greater than might be
® rst expected, however. Ixodes scapularis naturally harboured population s of the entomopa-
thogenic bacterium Bacillus thuringiensi s, albeit at nonlethal doses (Martin & Schmidtmann,
1998). In laborator y experiments, varroa mites feeding on A. mellifera adults and larvae
sprayed with the facultative insect pathogeni c bacterium, Serratia marcescens (used here as
a bioindicator) , ingested more than 10 7 bacteria per mite, 36 h after the start of feeding
(Glinski & Jarosz, 1990a).
Nematodes. Over 30 families of nematodes are associated with insects (Kaya & Stock,
1997). Of these, four families (Mermithidae, Allantonematidae, Steinernematidae, and
Heterorhabditidae ) have been studied for biologica l pest control (Hominick & Collins, 1997).
The Allantonematida e and Heterorhabditida e possess anatomical features that are used to
pierce or rupture the host cuticle; the other families infect through the spiracles, mouth, or
anus. Mermithid nematodes also infect hosts passively following the ingestion of their eggs
(Tanada & Kaya, 1993). Howardula acarinora (Allantonematidae ) is a highly adapted,
obligat e parasite of mesostigmatid mites of ® ve families, including an ectoparasite of bees,
Parasitus fucorum (Wachek, 1955; Siddiqi, 1985). Reproduction occurs outside the host and
only females are infective (P.N. Richardson, pers. comm.). However, hosts die slowly, if at
all, although they may be sterilized at high doses. In vitro economic rearing of Howardula
species is very unlikely (Richardson & Chanter, 1981), which further limits their potential
as microbial control agents. If H. acarinora were able to infect varroa then it might be
possible to introduce nematode-infected, mobile mesostigmatids (e.g. Stratiolaelaps miles)
into the honeybee colony and hope for cross-infection, although the practicalities of this
approach, and its likely eVectiveness, are unknown.
The nematode families Steinernematida e and Heterorhabditidae , (commonly termed
EPNsÐ entomopathogeni c nematodes) are highly virulent, obligate parasites of insects, and
are the most important nematodes for biological pest control. They introduce symbiotic,
pathogenic bacteria into the haemocoel of their hosts following penetration. Subsequent
multiplicatio n of the bacteria leads to host death within 48 h of infection. Up to 10
commercial EPN products have been availabl e in the UK (Richardson, 1996) and over 60
have been availabl e in Europe, mainly targeted against pestiferous Coleoptera, Lepidoptera
and Diptera, particularly for use in soil environments (P. N. Richardson, pers. comm). EPNs
can be mass produced, formulated and applied readily, and biopesticides based on EPNS
can be stored for extended periods. There are reports of the infectivity of EPNs to Acari,
notably in the Ixodida. In laborator y experiments, the cattle tick Boophilu s annulatu s was
susceptible to eight strains (six species) of Steinernema and nine strains (at least three
species) of Heterorhabditis, although the cattle ticks Boophilus microplus and A. variegatum
were resistant to infection (Mauleon et al., 1993). Strains of Heterorhabditis were generally
more virulent than Steinernema, but no nematodes reproduced in the ticks (Mauleon et al.,
1993). Steinernema carpocapsae caused > 90% mortality of B. annulatus in the laborator y
(Samish & Glazer, 1992; Glazer & Samish, 1993) and one strain of this nematode controlled
engorged female ticks in soil at economically feasible doses in glasshouse experiments
(Samish et al., 1999). Steinernema carpocapsae and Steinernema glaseri are also pathogeni c
to engorged adult I. scapularis (Zhioua et al., 1995). Steinernema feltiae has been shown
to kill Amblyomma americanum, Dermacentor variabilis and Rhipicephalu s sanguineus in
laborator y bioassays, while Steinernema riobrave killed D. variabilis, R. sanguineus, Ambly-
omma maculatum and Amblyomma cajenennse (Kocan et al., 1998).
The susceptibility of varroa and its close relatives to EPNs is not known but warrants
preliminary investigatio n because of the susceptibility of the Ixodidae. Species of EPN tend
to have broad host ranges, but varroa may present a physical barrier to infection. Varroa
mites spend much of their lives with their mouthparts connected to the host, leaving the
anus and the stigmata as the main ports of entry for nematodes. EPNs actively seek their
hosts, hence targeting adult varroa mites on bees, and larval mites in capped brood cells,
may be problematic. EPNs are adapted primarily to live in the soil and require high
humidities for infection (Kaya & Gaugler, 1993) although suitably moist conditions may
occur in some parts of the honeybee colony, e.g. brood cells. Species and strains of EPNs
diVer signi® cantly in their temperature optima, which probably re¯ ects their climate of
origin (Grewal et al., 1994). For example, the optimum temperature for activity of species
of Heterorhabditis from northwest Europe is 20± 25ë C (GriYn, 1993) whereas an isolate of
Heterorhabditis sp. from Israel was highly tolerant of temperatures above 30ë C (Glazer
et al., 1996) and hence could be expected to survive the temperatures of honeybee colonies
in summer. S. glaseri and S. riobrave can infect hosts at 37± 39ë C (Baur et al., 1995). These
nematodes caused only moderate brood loss when sprayed at high inoculum levels onto
A. mellifera combs (Baur et al., 1995). Honeybees are slightly susceptible to some other
species of EPN. Spraying S. carpocapsae infectives directly onto honeybee colony frames
containing A. mellifera gave no brood mortality but did result in up to 10% mortality of
adult bees (Kaya et al., 1982). Spraying adult bees in cages at 25ë C caused 15% mortality
(Kaya et al., 1982). Steinernema carpocapsae reproduced in bees in laborator y tests (Bailey,
1971) but is unlikely to do so at the temperature in the brood area (Kaya et al., 1982).
Protozoa. Approximately 1200 species of protozoa (Protista) are associated with insects
(Lipa, 1963). The entomopathogeni c protozoa are located in four phyla (Undeen & Vavra,
1997): Sarcomastigophor a (¯ agellates and amoebae); Apicomplexa (gregarines, neogregar-
ines, coccidia); Microspora (microsporidia) ; and Ciliophora (ciliates). Many of these species
have complex life-cycles, and both vertical and horizontal modes of transmission are
common. Horizontal infection is usually per os, although some ciliates are able to penetrate
their hosts through the cuticle. Those species causing chronic infections are often con® ned
to the host gut, whereas the more virulent taxa occupy intra- or intercellular localities. Host
mortality, when it occurs, is caused by irreparable damage to tissues and organs (Tanada &
Kaya, 1993). Of the various taxa, the microsporidia (Microspora) show the greatest promise
for biologica l pest control. These microorganisms are widespread in nature, and all are
obligat e intracellula r pathogens. They form the largest group of entomopathogeni c proto-
zoans, and many are highly virulent to their hosts. Unfortunately, they cannot be grown in
vitro, although cell culture is possible (Tanada & Kaya, 1993). Entomopathogeni c members
of the other protozoan phyla are frequently associated with chronic (sublethal) infections of
insects and, although common in nature, they are considered less suitable for biocontrol
(Tanada & Kaya, 1993).
Pathogenic protozoa have been reported from a range of acarine hosts. Gregarines
(Apicomplexa) have been recorded from the oribatids Damaeus oblongu s, Damaeus genicu-
latus, Scutovertex minutus, and the prostigmatid water mite Limnochares aquatica (Lipa,
1971). Helicosporidium parasiticum (unknown aYnity) attacks Hericia hericia (Astigmata )
a mite that lives in sap exuded from trees (Lipa, 1971). Mites infected by microsporidia
include Tyrophagus spp. (Astigmata), Acarus spp. (Astigmata), Neoseiulus [Amblyseius]
spp. (Mesostigmata) and certain Oribatidae (Lipa, 1971; Hazard et al., 1981; Bjornson et al.,
1996). Predatory phytoseiid mites (Mesostigmata), mass-reared for commercial biologica l
control, are sometimes adversely aVected by infections by microsporidia (Steiner & Bjornson,
1996). For example, Phytoseiulus persimilis showed reduced prey consumption and fecundity
when infected with Microsporidiu m phytoseiuli (Bjornson & Keddie, 1999). Gurleya sokolovi
(microsporidia ) is highly pathogeni c to L. aquatica (Issi & Lipa, 1968) and Nosema sperchoni
(microsporidia ) causes high levels of mortality in water mites of the genus Sperchon
(Prostigmata) (Lipa, 1962). Nosema may oVer the best prospect for Varroa control by a
protozoan. A number of species cause epizootics; Nosema locustae, for example, causes
natural epizootics in population s of acridids and is registered as a biological control agent
of rangeland grasshoppers in the USA (Shah & Goettel, 1999). Nosema species usually have
a high degree of host speci® city and it is likely that a species active against mites (e.g.
N. sperchoni) would not damage bees. The susceptibility of mesostigmatids to Nosema is not
known and warrants attention, although the mass production of any microsporidia n control
agent of Acari is likely to be problematic. However, the bee pathogen Nosema apis
demonstrates that it is possible for at least one Nosema species to operate under the physical
conditions of the honeybee colony. It may therefore be worth screening mite-active Nosema
species against varroa. Establishin g initial infections by microsporidia may be a challenge,
because the horizontal transmission of these pathogens depends on the ingestion of spores.
However, subsequent vertical transmission, if it occurs, could contribute to the sustained
control of varroa populations.
Viruses. Over 1600 viruses have been recorded from more than 1100 species of insects and
mites (Smits, 1997). Of these, three families (Baculoviridae, Polydn aviridae, Ascoviridae) are
speci® c for insects and related arthropods. The baculoviruse s are the most widely exploited
virus group for biocontrol (Martignoni , 1984). The mode of pathogenesi s and replication of
entomopathogeni c viruses varies according to the family, but infection nearly always occurs
by ingestion. Virions then bind to receptors in the gut and penetrate epithelial cells.
Infections of cytoplasmic polyhedrosi s viruses are generally con® ned to the midgut, while
infections of more virulent families, such as the baculoviruses, spread to the haemocoel and
then to essential organs and tissues, particularly fat bodies (Federici, 1993). Acute infections
lead to host death in 5± 14 days (Smits, 1997). Mass production can only be done in vivo,
but is economically viable for larger hosts such as Lepidoptera, and formulation and
applicatio n are straightforward. At present, there are approximatel y 16 biopesticides based
on baculoviruse s availabl e for use or under development. The majority of these products
are targeted against Lepidoptera (Shah & Goettel, 1999). The host range of baculoviruse s
is restricted to the order, and usually the family, of the host of origin, and commercial
baculoviru s biopesticides are considered non-harmful to A. mellifera (GroÈner, 1990).
There are very few reports of viral infections of Acari, and those viruses infecting
Mesostigmata are of low virulence. Mass production of a viral acaricide is likely to be a
challenge and no proprietary control agents have been developed. Three non-occluded
viruses are known to attack the tetranychid (Prostigmata) mites Panonychu s citri, Panonychus
ulmi and Tetranychus spp. (Lipa, 1971; Reed, 1981), and they sometimes reduce these pests
to very low numbers in the ® eld (Van der Geest, 1985). However, they are not known to
infect mesostigmatid mites (Beavers & Reed 1972; Reed 1981; Van der Geest 1985). The
following members of the Ixodida are described as sometimes suVering from a `presumed
virosis’ , but the authors consider these records should be treated with moderate scepticism:
B. microplus, Dermacentor marginatus, Argas persicus, Ornithodorus lahorensis, Ornithodorus
moubata, Ornithodorus tartakovskyi, Ornithodorus tholozani and Ornithodorus verrucosus
(Martignoni & Iwai, 1981). A non-occluded virus has been observed in P. persimilis
(Sutakova & Ruttgen, 1978; Steiner & Bjornson, 1996). A putative iridoviru s infection was
isolated from apparently healthy varroa mites associated with a decline in a honeybee colony
in the USA, although its pathogenicit y to the mites and bees was not determined (Camazine
& Liu, 1998). Virus-like particles, distinct from acute bee paralysis virus, were observed in
the fat body and muscle of varroa (Kleespies et al., 1994) and were associated with reduced
mite longevity, but could not be transmitted to healthy mites in preliminary experiments
(Kleespies et al., 2000).
Bacillus thuringiensi s (Bt). The majority of bacterial pathogen s of insects and related taxa
occur in the families Bacillaceae, Pseudomonadaceae, Enterobacteriaceae, Streptococcaceae,
and Micrococcaceae. Infections by the family Rickettsiaceae are dealt with in a later section
of this review. Most of these bacteria are weak pathogens that infect insects subject to
environmental stress, but a minority are highly virulent (Tanada & Kaya, 1993). By far the
most attention has been given to the Bacillaceae. Bacillus popillae causes milky disease in
scarbaeids, while Bacillus sphaericus is a lethal pathoge n of mosquitoes. Bacillus thuringiensi s
(Bt) is widespread in soil, is a lethal pathogen of a range of orders and is the most widely
used entomopathogeni c biologica l control agent. There are at present over 40 Bt products
availabl e for the control of caterpillars, beetles and small haematophagou s ¯ ies (Shah &
Goettel, 1999). Together, these products account for 1% of the world insecticide market
(Smits, 1997).
Bt is a spore forming bacterium. Sporulation is usually associated with the synthesis of a
proteinaceous protoxin crystal that has insecticidal activities, although some types of Bt
produce crystals with no known activity (Dulmage, 1981). Ingested crystals dissolve within
the gut and are cleaved by host proteases to form an active toxin, termed the d -endotoxin.
This binds to receptors in the midgut epithelium to cause the formation of ion pores, leading
to gut paralysis. The ingested spores may contribute to bacterial septicaemia. Host death
occurs quickly, often within a few days (Soares et al., 1994). Sixty seven serotypes (subspecies)
of Bt have been identi® ed, based on ¯ agellar antigens (Zeigler, 1999). Subspecies diVer in
host spectrum, but most show activities to Lepidoptera, Diptera and Coleoptera. Recent
searches have identi® ed isolates with activities against Hymenoptera, Homoptera and
Phthiroptera (Ellar, 1997). Novel toxins have also been identi® ed with activities against non
insect hosts, including some plant and animal-parasiti c nematodes, trematodes, protozoa,
and Acari (Feitelson et al., 1992). Some strains also produce exotoxins, which have a wide
spectrum of activity including vertebrates (Bailey, 1971). Bt products used for pest control
are normally based on exotoxin-negativ e strains. The b -exotoxin (thuringiensin ) is reported
to kill adults and larvae of the tetranychid mites Panonychu s citri (Hall et al., 1971),
Tetranychus paciWcus (Prostigmata) and the phytoseiid Metaseiulus occidentalis [Typhlod-
romus occidentalis] (Mesostigmata) (Hoy & Ouyang, 1987), plus the ectoparasitic northern
fowl mite, Ornithonyssu s sylviarum (Mesostigmata) (Mullens et al., 1988).
Various Bt strains had no eVect on the following mites species: (i) Prostigmata; Bryobia
arborea, Panonychus ulmi, Hydrachna sp., Aculus schlechtendali, Anystis sp.: (ii) Mesostig-
mata; Amblyseius potentillae and Typhlodromu s pyri (Lipa, 1971; Krieg & Langenbruch,
1981; Hassan et al., 1983, 1987; Van der Geest, 1985). However, Bt isolates reportedly lethal
to the two spotted spider mite, Tetranychus urticae (Prostigmata) have been patented in the
USA, together with a process for controlling mite pests of livestock, fowl and stored
products (Payne et al., 1993). Bt subsp. tenebrionis was moderately toxic to Metaseiulus
occidentalis under laborator y conditions (Chapman & Hoy, 1991), while Bt subsp. kurstaki
killed Ixodes scapularis in laborator y bioassays (Zhioua et al., 1999). Bt strains have been
isolated from varroa-infested honeybee colonies (Panizzi & Pinzauti, 1988) and from the
gut of varroa mites (Glinski & Jarosz, 1990b), but their pathogenicit y against the pest is not
known. Bioassays of 13 strains of Bt isolated from soil showed mortality of varroa to be ca.
15% higher than controls (Chang et al., 1993) and a strain of Bt tested in the former USSR
was reported to be toxic to varroa but practically harmless to honeybees (Mikityuk &
Korzhova, 1981; Mikityuk, 1984). Feitelson et al. (1992) considered there to be good
opportunitie s to ® nd Bt strains active agains t new targets, providin g the test organis m could
ingest and process Bt toxins and a suYciently diverse strain collection could be screened in
a viable bioassay. Extensive strain collections of Bt are availabl e that could be used for
screening agains t varroa (Donaldson, 1982; Dean & Zeigler, 1994; Cannon, 1996). There
may also be potential to develop a rapid screen for mite activity by probing for endotoxin
gene sequences associated with acaricidal activity (P. Jarrett, HRI Wellesbourne, pers.
comm.).
Most Bts have no adverse eVect on A. mellifera (Krieg & Langenbruch, 1981) and Bt is
accepted by regulatory authorities in the USA, the UK and some other countries as safe
for use in honeybee colonies (Vandenberg & Shimanuki , 1990). Bt operates eVectively at
high temperatures, including those experienced in honeybee colonies (see Burges, 1981).
High humidity could accelerate the rate of decay of Bt sprayed into the honeybee colony,
but this is unlikely to be a serious problem as Bt strains used against Galleria mellonella in
an active honeybee colony remained eVective for between ten weeks to one season (Vanden-
berg & Shimanuki, 1990). Bt should be considered, therefore, as a candidate for the
biocontrol of varroa. It could be sprayed onto bee clusters in winter, or onto brood chambers
in summer, but it should also be possible to use slow release application s from foundation
wax, a method that has been used against G. mellonella (Burges, 1976, 1977). Since the
eVectiveness of Bt is related to dose, the level of exposure that non-target mites would
receive outside the honeybee colony could well be non-harmful. If this was found not to be
the case, an alternative would be to engineer a Bt with a narrow host range. There have
been some signi® cant recent development s in modifying the activity and host spectrum of
naturally-occurrin g Bt strains (Cannon, 1996).
Rickettsiae. The rickettsiae are intracellular bacteria belonging to the alpha and gamma
groups of the Proteobacteria (Eubacteria) (Drancourt & Raoult, 1994). The genera Rickett-
sia, Rochalimea and Coxiella contain potent human pathogen s (Larsson, 1978) and can be
transmitted by ticks and mites (Burgdorfer & Varma, 1967). Members of the genus
Rickettsiella are pathogens of arthropods (Larsson, 1978) and although they are not related
antigenically to rickettsial pathogens of vertebrates, some can cause infection in the lungs
of mammals if inhaled (Tanada & Kaya, 1993). The Ricketsiella infect their arthropod hosts
per os and cannot be grown readily in bulk. They have been reported to cause infections
mainly in Coleoptera, Diptera and Lepidoptera (Tanada & Kaya, 1993). Vertical transmis-
sion occurs through the egg. Many entomogenous species of Rickettsiella cause sublethal
infections, but others are lethal, infecting the fat body and haemolymph (Sutakova &
Arutunyan, 1990). They also often exhibit low host speci® city (Larsson, 1978).
Rickettsial infections are well documented in the Insecta, but also in the Arachnida. Six
diseases caused by species of Rickettsiella and Porochlamydia have been described in spiders
and scorpions (Morel, 1978). Infections by rickettsiae and rickettsia-like organisms (RLOs)
have also been observed in the Acari, but are of low virulence. Laborator y population s of
M. occidentalis were susceptible to a deleterious RLO when reared in overcrowded conditions
(Hess & Hoy, 1982). Rickettsiae have been described in trombiculid mites (Prostigmata) and
in the predatory mite Phytoseiulus persimilis (Mesostigmata) (Hess & Hoy, 1982). Infections
by rickettsiae can signi® cantly aVect the longevity and fecundity of P. persimilis reared for
commercial biologica l control (Steiner & Bjornson, 1996). An unidenti® ed RLO, in all
stages of development, was also found in the rectum of varroa (Liu & Ritter, 1988). However,
given that arthropod-activ e rickettsiae pose a risk to humans, they should not be considered
for varroa control. Even if a mite-active strain were identi® ed that was considered safe to
use, it is likely to be of low virulence and there would be severe diYculties in mass
production.
Fungi. Approximately 750 species of fungi in 56 genera are reported from arthropods
(Hawksworth et al., 1995). Pathogen s of arthropods are represented in all four phyla of the
true fungi but most species reside in the Zygomycota, Ascomycota, and the Mitosporic
(imperfect) fungi. These groups also contain the most virulent fungal pathogens of insects
and mites, all of which are transmitted horizontally. The Entomophthorale s (Zygomycota )
contain some species that are obligat e pathogens and cause natural epizootics in a range of
agricultural pests, although a number of these species cannot be grown readily in vitro
(McCoy et al., 1988). Members of the Mitosporic entomopathogen s are associated less
commonly with natural epizootics, but they are popular choices for biopesticides because
they can be mass-produced and applied readily. Over 20 mycopesticides are availabl e
commercially, mainly for the management of Homoptera, Coleoptera, Lepidoptera, Diptera,
and Orthoptera (Shah & Goettel, 1999). The majority of products are based on the
Mitosporic fungus Beauveria bassiana, but mycopesticides are also availabl e from Metarhiz-
ium anisopliae, Paecilomyces fumosoroseus, and Verticillium lecanii. Naturalis TM (Thermo
Trilogy Corp.), a commercial mycopesticide based on B. bassiana, has been used to control
Tetranychus urticae on glasshouse roses (Wright & Kennedy, 1996).
The entomophthoralea n fungi and entomopathogeni c Ascomycetes exhibit ecomorpho-
logical adaptation s to particular hosts, whereas the Mitosporic entomopathogen s tend to be
less specialized (Samson et al., 1988). However, these species of Mitosporic fungi are
pleomorphic with respect to pathogenicity-relate d characteristics, and diVerent isolates from
the same species often vary in host range, pathogenicit y, and response to the external
environment (McCoy et al., 1988). Infection occurs almost exclusively by the attachment
and growth of spores through the host cuticle. Colonizatio n is necrotrophic, with the fungus
invadin g the haemocoel before rammifying throughou t the host. Tissue mortality is caused
by mechanical disruption, water loss, and the action of toxins. Death occurs within 4± 7 days
of infection, often followed by proli® c sporulation on the surface of the cadaver. Both
infection and sporulation require the presence of free water or high humidity, the lower
limit for the germination of spores in vitro being ca. 93% RH (Gillespie & Crawford, 1986).
In some cases, entomopathogeni c fungi are able to cause infections at seemingly lower
humidities than those required for germination in vitro, because the microclimate humidity
of the host is higher than ambient (Milner et al., 1997). Formulating the spores in oils, or
oil-in-water emulsions, also facilitates infections at lower humidities (Burges, 1999).
Many isolates of entomopathogeni c fungi require moderate temperatures (15± 27ë C) for
optimal infection (and hence would be more suitable for application to honeybee colonies
in the winter), although the maximum temperatures for growth vary extensively with isolate,
e.g. 33± 36ë C for V. lecanii, 33± 40ë C for Conidiobolu s coronatus, and more than 37ë C for
Beauveria spp. (Burges, 1981). The genus Metarhizium appears to exhibit the widest range
of temperatures for growth (Fargues et al., 1992), and isolates active at high temperatures
have been identi® ed. For example, an isolate of Metarhizium Xavoviride, originatin g from
Madagascar, grew optimally at 34ë C and had a maximum temperature for growth of 38ë C
(Welling et al., 1994). Heat-active strains of M. anisopliae have also been obtained that
germinate rapidly at 37ë C (McCammon & Rath, 1994).
A range of species from the Mitosporic fungi, Zygomycetes and some other taxa kill
Acari. Chandler et al. (2000) collated records of 58 species of fungi infecting at least 73
species of Acari, either naturally or in experiments. Fungal pathogens have been reported
to kill representatives of all three orders of the Actinotrichida, as well as the Ixodida and
Mesostigmata in the Anactinotrichida . Most reports concern infections in prostigmatid
mites (24 spp. of fungi from 45 hosts). Fungal pathogens of the Ixodida comprised
approximately 16 species of fungi from 10 host species, while the Mesostigmata recorded 19
species of fungi from 14 host species. Some species of Mitosporic entomopathogen s kill
insects and Acari (although it is not known whether these species contain Acari-speci® c
isolates). At least two species of fungiÐ Neozygites Xoridana (Entomophthorales ) and
Hirsutella thompsonii (Mitosporic fungi)Ð are speci® c to Acari. Neozygites Xoridana is a
speci® c pathogen of tetranychid mites and causes epizootics in T. urticae during the summer
in the southern USA (Van der Geest, 1985; Brandenburg & Kennedy, 1987). However it
cannot be grown in vitro and does not kill mesostigmatid mites. Hirsutella thompsonii is a
speci® c fungal pathogen of Acari and many strains derive from the tropics (McCoy, 1981).
It does not kill bees (Goettel et al., 1990). It is predominantly a pathogen of prostigmatid
mites (McCoy, 1981) although it has been observed also from the phytoseiid Typhlodromalu s
peregrinus (Mesostigmata) (McCoy, 1981) and Trachyuropod a coccinea (Mesostigmata)
(Balazy & Wisniewski, 1982). Hirsutella rostrata has also been observed infecting the
mesostigmatids Proctolaelaps sp. and Dendrolaelaps tetraspinosus (Balazy et al., 1987).
Hirsutella thompsonii has an important role in control of an eriophyoid pest, the citrus rust
mite Phyllocoptrut a oleivora (Allen et al., 1994) and has potential for control of the coconut
mite Eriophyes guerreronis (Lampedro & Luis-Rosas, 1989). Formulated as `Mycar’ , it has
been used for control of P. citri and P. oleivora on citrus in the USA, and on P. oleivora in
Australia (Rath, 1991), although the product is no longer availabl e. Isolates of H. thompsonii
can grow between 5 and 37ë C, and the fungus is considered safe to mammals (McCoy,
1981).
Balazy et al. (1987) reported 24 species of fungi (including six new species) infecting mites
collected from forest litter, soils, subcortical detritus, ant hills and birds. Fungi observed
included B. bassiana and V. lecanii infecting laelapid mites (Mesostigmata). This suggests
there is potential for discovering new species and strains of fungi with potential for the
control of varroa. Germinated conidia, of a fungus tentatively identi® ed as Aspergillus sp.,
have been observed penetrating the cuticle of varroa (Liu & Ritter, 1988). Some laborator y
studies have demonstrated that fungi will attack varroa, although high doses and extended
exposure were required. For example, when varroa mites were con® ned in Petri dishes for
1± 2 weeks with Aspergillus sp. and Beauveria sp. at 26ë C they became infected and died
1± 3 days later (Chernov, 1981).
Beauveria bassiana, M. anisopliae, Paecilomyces farinosus, P. fumosoroseus and V. lecanii
infect ixodid ticks in nature, and B. bassiana and M. anisopliae are being studied as biological
control agents of cattle ticks in Africa and South America. Metarhizium anisopliae and
B. bassiana, for example, have been observed infecting soil-dwelling stages of Boophilus
microplus in Brazil (Verissimo, 1995). Two isolates of M. anisopliae, originating from non-
acarine hosts, killed B. microplus in laborator y experiments (Bittencourt et al., 1992,
1994a,b) . Similarly, in Kenya, Rhipicephalu s appendiculatu s was susceptible in laborator y
bioassays to B. bassiana and M. anisopliae originatin g from non-acarine hosts (Mwangi
et al., 1995). These fungi subsequently caused up to 100% mortality of free living ticks in
the ® eld (Kaaya et al., 1996). A survey of natural mycoses in Ixodes ricinus in Denmark
(Kalsbeek et al., 1995) identi® ed six species of Mitosporic fungi (B. bassiana, Beauveria
brongniartii, P. farinosus, P. fumosoroseus, V. lecanii and Verticillium aranearum) infecting
diVerent life stages of this pest. Paecilomyces farinosus was the most frequently observed,
and infection rates of over 20% were recorded in engorged females.
There are records of B. bassiana, M. anisopliae, P. farinosus and V. lecanii infecting
A. mellifera, but no fungal control agent has been reported to cause an epizootic among
honeybees (Goettel et al., 1990). It should be noted that diVerent isolates of the same species
can have varying degrees of speci® city (Goettel et al., 1990). Beauveria bassiana, for example,
is known to infect more than 700 arthropod species (Goettel & Johnson, 1992) but exhibits
signi® cant pleomorphism. One strain of B. bassiana multiplied in honeybees in laborator y
tests, but has never been found attacking bees in nature (Bailey, 1971). The product Naturalis
(based on B. bassiana) had no signi® cant eVect on A. mellifera in a 30-day dietary and
contact study (Wright & Kennedy, 1996). Ball and Allen (1984) found that Vertalec (aphid-
active strain of V. lecanii), at 10 times the ® eld applicatio n rate, had no eVect on honeybees
at 25ë C in the laborator y, but Mycotal (white¯ y-active strain of V. lecanii) killed 15% of
bees and mycelial growth was observed in the thorax of aVected bees.
Entomopathogeni c fungi should be considered as prime candidates for screening against
varroa. Many species are active against Acari in nature, including mesostigmatid mites.
Hosts are infected through the cuticle, and there is evidence that the integument of varroa
can be breached. Several species of entomopathogeni c fungi are mass-produced and marketed
commercially and some of the knowledge and expertise associated with this may be
transferable to a mycopesticide for the control of varroa. A large resource-base of material

is also availabl e in culture collections for screening. Species of entomopathogeni c fungi vary
widely in pathogenicity-relate d characteristics, hence there should be opportunitie s to
identify a varroa-active fungus that operates at high temperatures and is not detrimental to
bees. Infection generally requires high humidities, but this can be overcome by formulation.
The humidity of the honeybee colony could also be raised temporarily with no ill eVect on
bees. Application s would be straightforwar d and could be made to honeybee colonies in
winter or summer. Fungal conidia could be applie d as a liquid spray onto bee clusters or
brood, or as a powder, possibly using a conidia transfer apparatus placed at the entrance of
the colony (Butt et al., 1998; Hatjina et al., 1998). Hirsutella thompsonii is a particularly
promising species to investigate. This fungus can be mass produced, it is speci® c to Acari,
some isolates infect mesostigmatid mites and it can grow at high temperatures.
FINAL ASSESSME NT
From this review, it is possible to rank natural enemy groups for their potential to control
varroa (Table 1). Entomopathogeni c fungi appear to have most potential, followed by Bt,
then protozoa and nematodes. Varroa can be kept alive in the laborator y on bee pupae
(B. Ball, unpublished ) or bee larvae in gelatine cells (Milani & Nazzi, 1994) and this could
form the basis of a laborator y bioassay of pathogen activity. Techniques are also availabl e
for enabling varroa to feed through synthetic membranes to screen gut-acting pathogens
(Bruce et al., 1991; Ball, 1994a).
A pathogen able to maintain varroa population s below a damage threshold would be
extremely valuable. High re-infestation rates may be expected in the early stages of a
biologica l control programme, especially if varroa is well-establishe d in feral population s or
if all the managed honeybee colonies in a locality are not treated together. The local spread
of varroa and the concomitant re-infestation pressure should not be underestimated (Powell,
1979; Ritter, 1988; Greatti et al., 1992; Mobus & De Bruyn, 1993; Walton, 1996). Treated
honeybee colonies can suVer from re-invasion rates of 40 to more than 100 mites per day
(Greatti et al., 1992; Fries, 1994; Martin, 1997).
A persistent pathoge n of varroa, used as a classical biologica l control agent, could spread
rapidly between managed colonies and into feral populations, giving long term or permanent
varroa control. In such a case it should also be speci® c to varroa, or non-targe t mite
population s could be at risk. An alternative, and probably more realistic, strategy would be
to use a nonpersistent pathogen with dose-dependent activity against varroa. This would
reduce the risk to nontarget mites outside the colony. Rapid and eYcient targeting techniques
should be possible with entomopathogeni c fungi and Bt, in which case biologica l control
would be an option for commercial and hobbyis t beekeepers alike. Finally, because the
TABLE 1. Ranking natural enemy groups for potential to control varroa
Lethality to the
Parasitiformes or
mites occupying Potential to
a comparable operate under
trophic niche to colony Ease of targeting Ease of mass Ease of
Group varroa conditions against varroa production registration
Predators/parasitoids 2 + 2 + ++
Nematodes + + 2 + ++
Protozoa + + + 2 +
Viruses 2 + 2 2 +
Bt + ++ + + +
Rickettsiae 2 + + 2 2 2
Fungi ++ + ++ + +
physical conditions inside honeybee colonies are similar everywhere, it is very likely that an
eYcient biocontrol agent of varroa could be used successfully throughout the world. This is
a rare opportunity in biologica l control.
ACKNOWLEDGEMENTS
This work was funded by the UK Ministry of Agriculture, Fisheries and Food (now the
Department for Environment, Food and Rural AVairs). IACR-Rothamsted receives grant-
aided support from the Biotechnolog y and Biological Sciences Research Council of the UK.
The authors wish to thank Dr. Mark Tatchell, Dr. Doreen Winstanley, Paul Jarrett, Paul
Richardson and Prof. Gwilym Evans for specialist advice and comment. Rhona Floate and
HRI Library staV provided invaluabl e assistance with literature searches.
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Biocontrol Science and

Prospective Biological Control

To cite this article: D. Chandler , K. D. Sunderland , B. V. Ball & G. Davidson (2001):

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Prospective Biological Control Agents of Varroa destructor

D. CHANDLER,1 K. D. SUNDERLAND,1 B. V. BALL2 and

Correspondence to: D. Chandler. E-mail: david.chandler@hri.ac.uk

and V. destructor n. sp have been separated on the basis of morphology, reproductive

SURVEY OF POTENTIAL BIOLOGICAL CONTROL AGENTS OF VARROA

parasitoid of a number of ticks of medical and veterinary importance. A French strain of

transferable to a mycopesticide for the control of varroa. A large resource-base of material

TABLE 1. Ranking natural enemy groups for potential to control varroa

W., Ed.), Ghent, Belgium, pp. 180± 182.

Trends in Ecology and Evolution 14, 312± 315.

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