and sporulation structures of Hirsutella vandergeesti (2)

Udział w badaniach nad grzybami entomopatogenicznymi<br />

Stanisław Bałazy, Instytut środowiska rolniczego i leśnego Polskiej Akademii Nauk, Bukowska 19, 60-809 Poznań; balazy@man.poznan.pl

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Beauveria eauve ia Gloeocystidium

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Research Centre for Agricultural <strong>and</strong> Forest Environment <strong>of</strong> the<br />

Polish Academy <strong>of</strong> Sciences (RCAFE), 60-809 Poznań, Bukowska<br />

str. 19, Pol<strong>and</strong><br />

Fungal diseases <strong>of</strong> invertebrates<br />

Stanisław Bałazy<br />

in decaying wood

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Entomophagous arthropods <strong>and</strong> pathogens<br />

connected with eight-toothed g bark beetle (Ips ( p typographus) yp g p ) <strong>and</strong> allied spruce p bark beetles<br />

1. Ovivorous mites<br />

Iponemus gaebleri<br />

I. balazyi<br />

Dendrolaelaps spp. <strong>and</strong> about 15 other<br />

gamasid species<br />

Pyemotes dryas<br />

2. Predacious insects<br />

About 15 bug <strong>and</strong> beetle species<br />

3. Parasitoids <strong>of</strong> adult beetles<br />

Rhopalophorus clavicornis<br />

Timicobia seitneri<br />

Mesopolobus typographi - hyperparasitoid<br />

Cosmophorus klugii<br />

C. regius<br />

Karpinskiella pityophthori<br />

4. A group <strong>of</strong> non-selective entomophagous insects<br />

About 20 species <strong>of</strong> Coleoptera <strong>and</strong> 15 <strong>of</strong><br />

Diptera, Heteroptera <strong>and</strong> Neuroptera.<br />

3. Parasitoids <strong>of</strong> larvae <strong>and</strong> pupae<br />

Coeloides bostrychorum<br />

Dendrosoter middendorfi<br />

Ecphylus hylesini<br />

Dinotiscus eupterus<br />

Eurytoma aethiops<br />

Eurytoma arctica<br />

Eurytoma spp.<br />

E. morio<br />

Gelis gentilis - hyperparasitoid<br />

Gelis sp. - hyperparasitoid<br />

Heydenia praetiosa<br />

Ipideurytoma spessivtsevi<br />

Metacolus unifasciatus<br />

Rhopalicus tutele<br />

Roptrocerus xylophagorum<br />

R. mirus<br />

Nasonia vitripennis<br />

Nemeritis transfuga<br />

Nemeritis sp.<br />

Neurateles papyraceus<br />

Serphus gravidator<br />

Proctotrupidae

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6. Pathogens <strong>of</strong> premature instars <strong>and</strong> adult beetles<br />

Bakteria - (poorly (p y recognized g –ca. 5 species) p )<br />

Fungi: Aphanocladium album<br />

Beauveria bassiana<br />

B. brongniartii<br />

Chytridiopsis typographi<br />

Conidiobolus coronatus<br />

H. cf. brownorum<br />

H. haptospora<br />

H. nodulosa<br />

<strong>Hirsutella</strong> rostrata<br />

<strong>Hirsutella</strong> sp.<br />

Isaria farinosa<br />

Lecanicillium muscarium<br />

Lecanicillium spp.<br />

Metarhizium anisopliae<br />

Si Simplicilium li ili spp.<br />

Tarichium spp.<br />

Nematoda:<br />

IIn adults: d lt<br />

Contortylenchus diplogaster<br />

Parasitorhabditis obtusa<br />

Polymorphotylenchus typographi<br />

SSulphuraetylenchus l h t l h sulfureus lf<br />

In larvae – invasive instars <strong>of</strong> unidentified species<br />

(about 15 species)<br />

<strong>Hirsutella</strong> cf. brownorum<br />

Protozoa<br />

GGregarina i ttypographi hi<br />

Nosema typographi<br />

Schizogragarinida (inidentified sp.)<br />

Unicaryon sp.

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Development <strong>of</strong> Neozygites abacaridis in the gall-mite Abacarus hystrix (1)<br />

<strong>and</strong> <strong>sporulation</strong> <strong>structures</strong> <strong>of</strong> <strong>Hirsutella</strong> v<strong>and</strong>ergeesti (2)<br />

1 2

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Entomophthora sp. cf. muscae<br />

Entomophaga tipulae<br />

Batkoa apiculata – resting spores<br />

Cordyceps ithacensis Paecilomyces tenuipes<br />

<strong>Hirsutella</strong> lecaniicola – anamorph <strong>of</strong> Cordyceps clavulata

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Nieokreślone przyczyny zamierania larw ogłodka brzozowca w Borach Tucholskich

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The old, ab<strong>and</strong>oned feeding sites become normally settled by consecutively changing saprotrophic<br />

nematode, t d mite it <strong>and</strong>d iinsectt communities, iti ussually ll ddominated i t d bby springtails, i t il sciarid i id llarvae <strong>and</strong>d gamasid id<br />

or tarsonemid mites, subject to the sawdust changes similar to over-fertilized organic soils. In such<br />

material the species <strong>Hirsutella</strong> minnesotensis originally occurring on nematodes in arable soils was<br />

found on juvenile j <strong>and</strong> adult tarsonemids.<br />

<strong>Hirsutella</strong> minnesotensis on tarsonemid female

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<strong>Hirsutella</strong> rostrata on Dendrolaelaps sp <strong>and</strong> on a predacious beetle larva<br />

<strong>Hirsutella</strong> rostrata on Dendrolaelaps sp. <strong>and</strong> on a predacious beetle larva<br />

Mycelium <strong>and</strong> Dictyochlamydospores

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A springtale infected by Conidiobolus coronatus<br />

<strong>Hirsutella</strong> sp. in proctotrupid wasp<br />

<strong>Hirsutella</strong> sp. on a braconid wasp<br />

(the arrows indicate the fungus synnemata)

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Contrary to the foli<strong>of</strong>agous <strong>and</strong> soil inhabiting noxious insects, epizootics caused by the entomopathogenic fungi in<br />

subcortical pest populations have been rather rare <strong>and</strong> never so spectacular. This can be caused by the strongly<br />

limited possibilities <strong>of</strong> direct artropod-pathogen contact, rather few infective units transmitted to subcortical<br />

habitats by parental cambio- or xylophagous hosts as well as by rather isolated their feeding in larval galleries.<br />

Entomophthora syrphi on Melanostoma scalare<br />

Furia sciare on sciariid flies P<strong>and</strong>ora neoaphidis on Microlophium carnosum

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Table 1. Selected d examples p s o<strong>of</strong> beneficial arthropod opod mycoses y os s (sp (species s<br />

linked with particular arthropod groups distinguished by boldface).<br />

Parasitoids - Hymenoptera Hymenoptera, Parasitica Parasitica Spiders - Arachnida<br />

Arachnida<br />

Batkoa major Acremonium spp.<br />

Beauveria bassiana Aphanocladium album<br />

<strong>Hirsutella</strong> in Serphus sp. Beauveria bassiana<br />

Paecilomyces farinosus B. brongniartii<br />

P<strong>and</strong>ora dacnusae Gibellula leiopus<br />

Zoophthora ichneumonis G. pulchra<br />

Zoophthora Zoophthora cf. cf. lanceolata lanceolata Hymenostilbe Hymenostilbe aranearum<br />

aranearum<br />

Verticillium lecanii <strong>Hirsutella</strong> aranearum<br />

Lecanicillium aranearum<br />

Ants – Hymenoptera, Formicoidea Lecanicillium spp.<br />

Aegeritella i ll spp.<br />

Batkoa apiculata<br />

Paecilomyces l farinosus f<br />

Beauveria bassiana Harvest spiders - Opiliones<br />

Cordyceps Cordyceps myrmecophila myrmecophila B. B. bassiana bassiana<br />

P<strong>and</strong>ora myrmecophaga Entomophaga batkoi<br />

Lecanicillium lamellicola<br />

Paecilomyces farinosus<br />

Metarhizium Metarhizium anisoplia anisoplia<br />

P<strong>and</strong>ora phalangicida

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• Ground beetles - Coleoptera, Carabidae Predaceous mites - Acari<br />

• Beauveria bassiana Acremonium sp<br />

• Cordyceps entomorrhiza Aphanocladium album<br />

• <strong>Hirsutella</strong> cf. eleutheratorum Beauveria bassiana<br />

• Lecanicillium cf. lecanii B. brongniartii<br />

• Metarhizium anisopliae Conidiobolus chlapowskii<br />

• Paecilomyces farinosus C. coronatus<br />

• <strong>Hirsutella</strong> brownorum<br />

• Hgregis H. gregis<br />

• Other predaceous beetles H. haptospora<br />

• Beauveria bassiana H. cf. minnesotensis<br />

• B. brongniartii H. nodulosa<br />

• <strong>Hirsutella</strong> sp. I H. rostrata<br />

• <strong>Hirsutella</strong> sp. II Tarichium sp. about 10 species<br />

• Lecanicillium spp. Lecanicillium lamellicola<br />

• Metarhizium anisopliae LL. lecanii<br />

• Paecilomyces farinosus L. psalliotae<br />

• P. fumosoroseus<br />

• P. tenuipes<br />

• Paecilomyces in Lampyris

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Gibellula pulchra on a spider

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Eryniopsis caroliniana<br />

<strong>Hirsutella</strong> aphidis<br />

<strong>Hirsutella</strong> acridiorum<br />

Entomophaga tenthredinis<br />

Batkoa apiculata

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Akanthomyces aculeata<br />

na motylu<br />

Hymenostilbe sp.<br />

na pająku<br />

Nieokreślony gatunek na Pterostichus sp.<br />

EErynia i rhizospora hi na chruścikach h ś ik h

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Isaria dubia<br />

CC. gracilis<br />

ili<br />

Tilachlidiopsis p nigra g – C. entomorhizza<br />

Cordyceps militaris<br />

Maczużniki<br />

(Cordyceps)<br />

ii i ich h<br />

anamorfy

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Cordyceps gracilis <strong>and</strong> the synnemata<br />

<strong>of</strong> its anamorph (Paraisaria dubia)<br />

Cordyceps clavulata <strong>and</strong><br />

<strong>Hirsutella</strong> lecaniicola

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Morrtality<br />

(% )<br />

epizootic level<br />

enzootic level<br />

Months<br />

- .- .- .- E.batkoi on harvest-spiders Oligolophus tridens.<br />

--------- Zoophthora bialoviezaensis on foliophagous geοmetrid caterpillars.<br />

xoxoxo Entomophthora culicis on chironomid adults in l<strong>and</strong>-water ecotones.<br />

- .- .- .- Ζ. arginis g on larvae <strong>of</strong> Arge g berberidis.<br />

……... ΕΕ. muscae on adult calyptrate flies. flies<br />

--------- Ζ. aphrophorae on Dicyphus pallidus.<br />

……... Conidiobolus obscurus on different aphid species.<br />

xxxxxx Ζ. rαdicans on Pieris brassicae caterpillars.<br />

xxxxxx Neozygites floridana on Tetranychus urticae.<br />

oooooo Ζ. phytonomi on Phytonomus spp. larvae in alfalfa.<br />

oooooo Tarichium rhagonycharum on adults <strong>of</strong> Rhagonycha lignosa in forests.<br />

xxxxxx Erynia conica on Chironomidae, Culicidae allied gnats.<br />

- .- .- .- Cordyceps entomorrhiza on larvae, pupae or adults <strong>of</strong> Carabus spp.<br />

- .- .- .- P<strong>and</strong>ora neoaphidis on different aphid species.<br />

______ Beauveria bassiana on different species in forest litter. litter<br />

xoxoxo P<strong>and</strong>ora echinospora on adult laιιxaniid flies.<br />

______ Paecilomyces farinosus on larvae <strong>and</strong> pupae <strong>of</strong> Microlepidoptera.<br />

xoxoxo Furia sciarae on adult sciarids in moist deciduous forest undergrowth.<br />

--------- Lecanicillium spp. on Aphidoidea <strong>and</strong> Orthezia urticae.<br />

……... Entomophaga tenthredinis on tenthredinid larvae.<br />

oooooo Gibellula pulchra on spiders in moist grassl<strong>and</strong>s

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Numbers <strong>of</strong> anemoconidia <strong>of</strong> Beauveria bassiana <strong>and</strong> Paecilomyces<br />

farinosus (in millions per 1m2 ) estimated on the basis <strong>of</strong> dead insects found<br />

on research plots (different letters indicate statistically significant values,<br />

the asterisks (*) – highly significant; Whitney-Mann U-test)<br />

Habitat Pologne Romania France<br />

Forests 711,3 A� 1224,1 A� 176,8 A�<br />

Mid-field afforestations 116,3 B 636,3 B 190,2 A�<br />

Perennial crops <strong>and</strong> grassl<strong>and</strong>s 112,6 B 624,2 1) B 161,0 A�<br />

Annual crops 55C� 5,5 C� 83 83,2 2 B� B� 08B 0,8 B<br />

1) only y<br />

alfalfa cultures.

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Almost all entomophthoralean species affecting holometabolic insects have<br />

been specialized to infect only their one – larval by or adult - development<br />

instar.<br />

The larval mortality, caused mostly by the pathogens <strong>of</strong> the genera<br />

Entomophaga <strong>and</strong> Zoophthora, is commonly <strong>of</strong> great importance for the plant<br />

protection t ti bboth th iin agriculture i lt <strong>and</strong>d fforestry. t<br />

On the other h<strong>and</strong>, frequent cases <strong>of</strong> very high mortality <strong>of</strong> adult anthomyid<br />

flies caused by Entomophthora muscae do not reduce significantly their<br />

populations l ti bbecause <strong>of</strong>f hhost’ t’ddying i after ft oviposition i iti period. i d<br />

Dipteran larvae generally appear to be resistant to infection both by the<br />

entomophthoralean <strong>and</strong> mitosporic fungi.<br />

Heavy epizootics caused in moist bushy or forest habitats by Paecilomyces<br />

suffultus in gregarious bibionid larvae were not frequent exceptions. This<br />

fungus merits to be tested for the pathogenicity to other dipteran larvae, as a<br />

potential supplementary agent agent.

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Micromorphology <strong>of</strong> some anamorphs<br />

Paecilomyces farinosus<br />

Gibellula<br />

<strong>Hirsutella</strong><br />

Beauveria<br />

Nomuraea<br />

Lecanicillium

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Identification <strong>of</strong> <strong>Hirsutella</strong>-species is generally difficult because <strong>of</strong> the<br />

scarcity <strong>of</strong> differentiating features <strong>and</strong> very great similarity <strong>of</strong> morphological<br />

elements bearing taxonomical value. Some <strong>of</strong> them are able to infect both insects<br />

<strong>and</strong>d mites it or nematodes t d <strong>and</strong>d mites, it as ffor iinstance t HH. nodulosa d l <strong>and</strong>d HH. minnesotensis<br />

i t i<br />

respectively. Lately also H. rostrata – known only from mites – was found on a ciidbeetle<br />

larva.<br />

Neither strains <strong>of</strong> H. thompsonii, H. minnesotensis or H. rostrata infected<br />

insects in laboratory assays (caterpillars <strong>of</strong> the codling moth <strong>and</strong> larve <strong>of</strong> birch barkbeetle)<br />

whereas H. nodulosa caused single positive cases in the later.<br />

Though by biotests are necessary to check the strain pathogenicity to the<br />

other th gropus iinvertebrates, t b t th the main i diffi difficulty lt iin th the llaboratory b t bi bioassays with ith some<br />

<strong>Hirsutella</strong> strains is to obtain a sufficient amount <strong>of</strong> spores for required<br />

concentration <strong>of</strong> infective suspensions<br />

(e. ( g. g the strain 3747 m – <strong>of</strong> H. cf. brownorum sporulates p extremelyy weakly). y)<br />

H. minnesotensis shows very close morphological similarity to H.<br />

thompsonii but the DNA ITS region sequencing <strong>of</strong> its strains from tarsonemid mites<br />

excludes their identity, whereas the sequencies <strong>of</strong> its strains from insects <strong>and</strong><br />

ttarsonemid id mites it did nott diff differ.<br />

Several further specimens <strong>and</strong> cultures <strong>of</strong> <strong>Hirsutella</strong> from hitherto unknown<br />

insect <strong>and</strong> mite hosts are currently investgated in order to correct idebtification.

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Though the first fungous diseases <strong>of</strong> mites have been recognized in the<br />

turn <strong>of</strong> XIXth century deepened studies on them were concentrated mostly in<br />

regions, where phytophagous groups <strong>of</strong> mites constitute real problems in<br />

plant pathology. In Pol<strong>and</strong> first reports on hyphomycetous <strong>and</strong><br />

entomophthoralean diseases in mites appeared only in the second half <strong>of</strong><br />

XXth century y (Bałazy ( y 1970, , Bałazyy <strong>and</strong> Wiśniewski 1984, , Bałazyy et al. 1986). )<br />

By the end <strong>of</strong> 1990s co-ordinated research projects concerning occurrence<br />

<strong>and</strong> significance od mycoses in different groups <strong>of</strong> phytophagous,<br />

saprotrophic <strong>and</strong> beneficial mites were undertaken (Miętkiewski et al. 2000).<br />

Till now mycoses <strong>of</strong> two groups <strong>of</strong> mites have been recognized:<br />

(1) - phytophagous spider- <strong>and</strong> gall-mites in gardens, orchards <strong>and</strong> grassl<strong>and</strong>s;<br />

(2) - associations <strong>of</strong> saprotrophic <strong>and</strong> predacious mites occurring commonly in<br />

subcortical b ti l iinsect t ffeeding di grounds d ontrees.<br />

t

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Differentiation <strong>of</strong> H. thompsonii strains on the same medium

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Within the hyphomycetes <strong>Hirsutella</strong> <strong>and</strong> Lecanicillium anamorphs show<br />

very great variability <strong>and</strong> limited number <strong>of</strong> the morphological features,<br />

that would merit the value <strong>of</strong> criteria for their taxonomy. Modern<br />

approach based on DNA sequencioning <strong>of</strong> ITS region allowed to select<br />

from the collected strains some separated p groups. g p<br />

+NEMATODES<br />

MITES<br />

MITES<br />

NEMATODES<br />

MITES