Hirsutella thompsonii and Metarhizium anisopliae as potential microbial control agents of Varroa destructor,a honey bee parasite

The potential of Hirsutella thompsonii Fisher and Metarhizium anisopliae (Metschinkoff) as biological control agents of the parasitic mite, Varroa destructor Anderson and Trueman was evaluated in the laboratory and in observation hives. In the laboratory, time required for 90% cumulative mortality of mites (LT(90)) was 4.16 (3.98-4.42) days for H. thompsonii and 5.85 (5.48-7.43) days for M. anisopliae at 1.1 x 10(3) conidia mm(-2). At a temperature (34+/-1 degrees C) similar to that of the broodnest in a honey bee colony, Apis mellifera L., H. thompsonii [LC(90)=9.90 x 10(1) (5.86-19.35) conidia mm(-2) at Day 7] and M. anisopliae [LC(90)=7.13 x 10(3) (2.80-23.45) conidia mm(-2) at Day 7] both showed significant virulence against V. destructor. The applications of H. thompsonii to observation hives resulted in significant mortality of mites, and reduction of the number of mites per bee 21 and 42 days post-treatments. The treatments did not significantly affect the mite population in sealed brood. However, the fungus must have persisted because infected mites were still observed [82.97+/-(0.6)%] 42 days post-treatment. In addition, the fungus was found to sporulate on the host. A small percentage [2.86+/-(0.2)%] of dead mites found in the control hives also showed fungal infection, suggesting that adult bees drifted between hives and disseminated the fungus. H. thompsonii was harmless to the honey bees at the concentrations applied and did not have any deleterious effects on the fecundity of the queens. Microbial control with fungal pathogens provides promising new avenues for control of V. destructor and could be a useful component of an integrated pest management program for the honey bee industry.     

Insights into fungal communities colonizing the acarosphere in a forest soil habitat

Knowledge on the diversity and ecology of microfungi associated with soil-dwelling mites is rather limited. To get insights into associations between the two highly diverse groups, we studied composition and potential function of mite-associated fungal communities occurring in soil. Two different mite species living in temperate region pine forest soil were screened for associated fungi. The fungal community was assessed by restriction fragment length polymorphism (RFLP) analyses in a predatory (Leptogamasus obesus) and a predominantly saprobic (Oppiella subpectinata) mite species as well as in the organic soil layer. Key fungi were identified by sequencing, and community composition was exemplarily compared between the RFLP and a 454 metabarcoding approach. Composition of the fungal communities differed between mite species and between mites and organic soil layer. The mites were predominantly associated with Zygomycota, less frequently with Ascomycota, and rarely with Basidiomycota. The bulk soil was colonized by roughly equal proportions of the three phyla. Fungal taxa being known to exhibit chitinolytic activity were predominantly restricted to mites. Compositional and functional differences between the communities suggest that mites represent a particular microhabitat for fungi, the “acarosphere.” This mobile habitat may contribute to nutrient cycling by combining fungal and animal decomposition activities and serve as vector for soil-inhabiting fungi.     

Use of real-time PCR to discriminate parasitic and saprophagous behaviour by nematophagous fungi

Entomopathogenic nematodes (EPNs) are important pathogens of soilborne insects and are sometimes developed commercially to manage insect pests. Numerous nematophagous fungal species (NF) prey on nematodes and are thought to be important in regulating natural or introduced EPN populations. However, nematophagy by these fungi in nature cannot be inferred using existing methods to estimate their abundance in soil because many of these fungi are saprophytes, resorting to parasitism primarily when certain nutrients are limiting. Therefore, we developed an assay to quantify NF DNA in samples of nematodes. Species-specific primers and TaqMan probes were designed from the ITS rDNA regions of Arthrobotrys dactyloides, Arthrobotrys oligospora, Arthrobotrys musiformis, Gamsylella gephyropagum and Catenaria sp. When tested against 23 non-target fungi, the TaqMan real-time PCR assay provided sensitive and target-specific quantification over a linear range. The amount of A. dactyloides or Catenaria sp. DNA in 20 infected nematodes, measured by real-time PCR, differed between fungal species (P=0.001), but not between experiments (P>0.05). However, estimates of relative NF parasitism using a bioassay with 20 nematodes infected by either species, differed greatly (P<0.001) depending on whether the fungi were alone or combined in the samples used in the assay. Tests done to simulate detection of NF DNA in environmental samples showed that, for all species, background genomic DNA and/or soil contaminants reduced the quantity of DNA detected. Nested PCR was ineffective for increasing the detection of NF in environmental samples. Indeed, real-time PCR detected higher amounts of NF DNA than did nested PCR. The spatial patterns of NF parasitism in a citrus orchard were derived using real-time PCR and samples of nematodes extracted from soil. The parasitism by Catenaria sp. was positively related to the abundance of both heterorhabditid and steinernematid EPNs. The possible significance of the associations is ambiguous because NF attack a broad range of nematode taxa whereas EPNs are a small minority of the total nematode population in a soil sample. These studies demonstrate the potential of real-time PCR to study the role of NF parasitism in soil food webs.     

Virulence and site of infection of the fungus,Hirsutella thompsonii,to the honey bee ectoparasitic mite,Varroa destructor

The Varroa mite, Varroa destructor, is recognized as the most serious pest of both managed and feral Western honey bee (Apis mellifera) in the world. The mite has developed resistance to fluvalinate, an acaricide used to control it in beehives, and fluvalinate residues have been found in the beeswax, necessitating an urgent need to find alternative control measures to suppress this pest. Accordingly, we investigated the possibility of using the fungus, Hirsutella thompsonii, as a biocontrol agent of the Varroa mite. Among the 9 isolates of H. thompsonii obtained from the University of Florida and the USDA, only the 3 USDA isolates (ARSEF 257, 1947 and 3323) were infectious to the Varroa mite in laboratory tests. The mite became infected when it was allowed to walk on a sporulating H. thompsonii culture for 5 min. Scanning electron micrographs revealed that the membranous arolium of the mite leg sucker is the focus of infection where the fungal conidia adhered and germinated. The infected mites died from mycosis, with the lethal times to kill 50% (LT(50)s) dependent on the fungal isolates. Thus, the LT(50)s were 52.7, 77.2, and 96.7h for isolates 3323, 257, and 1947, respectively. Passage of H. thompsonii through Varroa mite three times significantly reduced the LT(50)s of isolates 257 and 1947 (P<0.05) but not the LT(50) of isolate 3323.The fungus did not infect the honey bee in larval, prepupal, pupal, and adult stages under our laboratory rearing conditions. Our encouraging results suggest that some isolates of H. thompsonii have the potential to be developed as a biocontrol agent for V. destructor. However, fungal infectivity against the mites under beehive conditions needs to be studied before any conclusion can be made.     

云南省锡金小鼠体表革螨感染分析

为了解云南省锡金小鼠(Mus pahari)体表革螨的感染情况及分布规律,本研究基于1990至2015年云南省39个县(市)的调查数据,统计分析锡金小鼠体表革螨的基本感染情况和感染差异.使用聚块指数测定革螨空间分布型,用Jaccard指数计算革螨物种相似性.从捕获的720只锡金小鼠体表共采集到革螨14 098只,鉴定为...     

A scientific note on the detection of honeybee viruses using real-time PCR (TaqMan) in Varroa mites collected from a Thai honeybee (Apis mellifera) apiary

Bee parasitic mite syndrome is a disease complex of colonies simultaneously infested with Varroa destructor mites and infected with viruses and accompanied by high mortality. By using real-time PCR (TaqMan), five out of seven bee viruses were detected in mite samples (V. destructor) collected from Thailand. Moreover, the results of this study provide an evidence for the co-existence of several bee viruses in a single mite. This is also the first report of bee viruses in mites from Thailand.     

Efficacy of strips coated with Metarhizium anisopliae for control of Varroa destructor (Acari: Varroidae) in honey bee colonies in Texas and Florida

Strips coated with conidia of Metarhizium anisopliae (Metschinkoff; Deuteromycetes: Hyphomycetes) to control the parasitic mite, Varroa destructor (Anderson and Trueman) in colonies of honey bees, Apis mellifera (Hymenoptera: Apidae) were compared against the miticide, tau-fluvalinate (Apistan) in field trials in Texas and Florida (USA). Apistan and the fungal treatments resulted in successful control of mite populations in both locations. At the end of the 42-day period of the experiment in Texas, the number of mites per bee was reduced by 69-fold in bee hives treated with Apistan and 25-fold in hives treated with the fungus; however mite infestations increased by 1.3-fold in the control bee hives. Similarly, the number of mites in sealed brood was 13-fold and 3.6-fold higher in the control bee hives than in those treated with Apistan and with the fungus, respectively. Like the miticide Apistan, the fungal treatments provided a significant reduction of mite populations at the end of the experimental period. The data from the broodless colonies treated with the fungus indicated that optimum mite control could be achieved when no brood is being produced, or when brood production is low, such as in the early spring or late fall. In established colonies in Florida, honey bee colony development did not increase under either Apistan or fungal treatments at the end of the experimental period, suggesting that other factors (queen health, food source, food availability) play some major role in the growth of bee colonies. Overall, microbial control of Varroa mites with fungal pathogens could be a useful component of an integrated pest management program for the honey bee industry.     

中国云南小兽寄生革螨的群落结构和空间分布

寄生在小兽体表的革螨可能是传播肾综合征出血热和立克次体痘等人兽共患病的媒介。本文报道了云南省28个县(市)小兽体表革螨的野外调查结果, 运用物种数、平均丰富度和Shannon多样性指数对小兽寄生革螨的群落特征和沿环境梯度的空间分布进行了研究, 并用系统聚类分析法 (SPSS 16.0软件)对18种主要小兽的革螨群落相似性进行了比较。在云南省28个县(市)共捕获到小兽14 544头, 隶属于5目（啮齿目、食虫目、攀鼩目、兔形目、食肉目）10科35属67种, 在捕获的小兽体表采集到革螨80 791头, 经鉴定属于10科33属112种。结果分析表明: 主要的宿主动物为黄胸鼠 Rattus tanezumi、齐氏姬鼠Apodemus chevrieri和大绒鼠Eothenomys miletus;纳氏厉螨Laelaps nuttalli、毒厉螨L. echidninus和贵州厉螨L. guizhouensis为革螨的优势种。齐氏姬鼠A. chevrieri、社鼠N. confucianus和黄胸鼠R. tanezumi体表寄生的革螨种类最多;臭鼩鼱Suncus murinus、齐氏姬鼠A. chevrieri和灰麝鼩Crocidura attenuata的革螨群落多样性最高。聚类分析结果表明, 大部分革螨群落的相似程度与相应小兽的亲缘关系及所处生态环境的相似性是基本一致的, 分类地位和生境选择相似的小兽, 它们的革螨群落也被聚为一类。革螨物种数沿纬度梯度的水平分布呈现两个峰值, 最大峰值出现在25°~26°N之间;沿海拔梯度的垂直分布呈单峰分布格局, 峰值在海拔2 000~2 500 m之间。革螨和小兽的多样性沿纬度梯度和海拔梯度的空间分布趋势也均表现出随着纬度和海拔的升高而先升高后降低的单峰型分布格局, 峰值分别出现在北纬25°~27°N和海拔2 000~2 500 m之间。结果提示云南革螨群落种类丰富, 多样性高。云南革螨物种数和多样性的分布格局可能直接受到古北和东洋两区系边缘效应的影响。     

Mites and fungi in heavily infested stores in the Czech Republic

Toxigenic and allergen-producing fungi represent a serious hazard to human food and animal feed safety. Ninety-four fungal species were isolated from mite-infested samples of seeds taken from Czech seed stores. Fungi were isolated from the surface of four kinds of seeds (wheat, poppy, lettuce, and mustard) and from the gut and external surface of five species of mites (i.e., Acarus siro L., 1758, Caloglyphus rhizoglyphoides (Zachvatkin, 1973), Lepidoglyphus destructor (Schrank, 1781), Tyrophagus putrescentnae (Schrank, 1781) and Cheyletus malaccensis Oudemans 1903) separately. Multivariate analysis of fungi complex composition showed that the frequency of fungal was species significantly influenced by the kind of seed. Fungal frequencies differed between mites gut and exoskeleton surface and between the surfaces of mites and seeds. Three groups of fungal species were recognized: 1) mite surface-associated fungi: Penicillium brevicompactum, Alternaria alternata, and Aspergillus versicolor; 2) mite surface- and seed-associated fungi: Aspergillus niger, Penicillium crustosum, Penicillium aurantiogriseum, Penicillium chrysogenum, and Aspergillus flavus; and 3) seed-associated fungi: Cladosporium herbarum, Mucor dimorphosporus f. dimorphosporus, Botrytis cinerea, Penicillium griseofulvum, and Eurotium repens. Mite-carried species of microfungi are known to produce serious mycotoxins (e.g., aflatoxin B1, cyclopiazonic acid, sterigmatocystin, ochratoxin A, and nephrotoxic glycopeptides) as well as allergen producers (e.g., A. alternata and P. brevicompactum). Storage mites may play an important role in the spread of some medically hazardous micromycetes. In addition, these mite-fungi associations may heighten the risk of occurrence of mycotoxins in food and feed stuffs and cause mixed contamination by fungal and mite allergens.     

Diversity of acaropathogenic fungi in Poland and other European countries

The occurrence, species diversity and some aspects of taxonomical affinity and host selectivity of acaropathogenic fungi associated with phytophagous, saprotrophic and predacious mites in Poland and other European countries were investigated on wild and cultivated plants, in insect feeding sites under the bark and in decayed wood. From among 33 species of fungi affecting mites only five species of Entomophthorales were separated and the most numerous were Neozygites floridana mostly on Tetranychus urticae, N. abacaridis on a few eriophyid species, and Conidiobolus coronatus attacking gamasid mites most frequently of the genus Dendrolaelaps. The most frequent mite pathogens occurring in mite communities on plants and in wood infested by insects were of the genus Hirsutella. Until now 13 of their form-species have been recognized in these habitats, but only H. kirchneri, H. necatrix and H. thompsonii (including its variety synnematosa) can be treated as exclusive oligophagous pathogens of phytophagous mites, though their potential host range seems to embrace only selected eriophyid or tarsonemid mites. Taxonomical differentiation of fungal strains was based on close morphological observations and molecular analysis of ITS region sequences. Two new species of acaropathogenic fungi were described in these studies. Hirsutella danubiensis sp. nov. was found in the tetranychid T. urticae, whereas H. vandergeesti sp. nov. affected phytoseiid mites of the genera Amblyseius, Neoseiulus, Seiulus and Typhlodromus, and the tarsonemid Tarsonemus lacustris.     

Diseases of Mites

An overview is given of studies on diseases of mites. Knowledge of diseases of mites is still fragmentary but in recent years more attention has been paid to acaropathogens, often because of the economic importance of many mite species. Most research on mite pathogens concerns studies on fungal pathogens of eriophyoids and spider mites especially. These fungi often play an important role in the regulation of natural mite populations and are sometimes able to decimate populations of phytophagous mites. Studies are being conducted to develop some of these fungi as commercial acaricides.Virus diseases are known in only a few mites, namely, the citrus red mite and the European red mite. In both cases, non-occluded viruses play an important role in the regulation of mite populations in citrus and peach orchards, respectively, but application of these viruses as biological control agents does not seem feasible. A putative iridovirus has been observed in association with Varroa mites in moribund honeybee colonies. The virus is probably also pathogenic for honeybees and may be transmitted to them through this parasitic mite.Few bacteria have been reported as pathogens of the Acari but in recent years research has been concentrated on intracellular organisms such as Wolbachia that may cause distorted sex ratios in offspring and incompatibility between populations. The role of these organisms in natural populations of spider mites is in particular discussed. The effect of Bacillus thuringiensis on mites is also treated in this review, although its mode of action in arthropods is mainly due to the presence of toxins and it is, therefore, not considered to be a pathogen in the true sense of the word.Microsporidia have been observed in several mite species especially in oribatid mites, although other groups of mites may also be affected. In recent years, Microsporidia infections in Phytoseiidae have received considerable attention, as they are often found in mass rearings of beneficial arthropods. They affect the efficacy of these predators as biological control agent of insect and mite pests. Microsporidia do not seem to have potential for biological control of mites.     

中国云南小兽体表革螨的群落相似性

运用系统聚类分析方法对中国云南省境内17种主要小型哺乳动物(小兽)体表革螨群落相似性进行研究,每一种小兽体表的所有外寄生革螨被定义为一个相应的革螨群落。运用SPSS11.5软件完成17种革螨群落的相似性比较。研究结果表明:小兽体表革螨群落结构复杂,物种多样性高;隶属同一个属的小兽体表的革螨群落相似程度高,在系统聚类分析中聚为一类;大多数革螨群落相似性大小与相应小兽宿主在动物分类上的近缘性高低呈现高度一致,但也有一些革螨群落是例外的。这说明小兽体表革螨群落不仅受小兽宿主分类地位的影响,可能还受宿主生境的影响     

Oribatid mite (Acari, Oribatida) feeding on ectomycorrhizal fungi

The coexistence of a large number of soil animals without extensive niche differentiation is one of the great riddles in soil biology. The main aim of this study was to explore the importance of partitioning of food resources for the high diversity of micro-arthropods in soil. In addition, we investigated if ectomycorrhizal fungi are preferentially consumed compared to saprotrophic fungi. Until today, ectomycorrhizal fungi have never been tested as potential food resource for oribatid mites. We offered six ectomycorrhizal fungi [Amanita muscaria (L.) Hook., Boletus badius (Fr.) Fr., Cenococcum geophilum Fr., Laccaria laccata (Scop.) Fr., Paxillus involutus (Batsch) Fr. and Piloderma croceum J. Erikss. & Hjortstam], one ericoid mycorrhizal fungus [Hymenoscyphus ericae (D.J. Read) Korf & Kernan] and three saprotrophic fungi [Agrocybe gibberosa (Fr.) Fayod, Alternaria alternata (Fr.) Keissl. and Mortierella ramanniana (A. Møller) Linnem.] simultaneously to each of the mainly mycophagous oribatid mite species Carabodes femoralis (Nicolet), Nothrus silvestris Nicolet and Oribatula tibialis Nicolet. The ericoid mycorrhizal fungus H. ericae and the ectomycorrhizal fungus B. badius were preferentially consumed by each oribatid mite species. However, feeding preferences differed significantly between the three species, with O. tibialis being most selective. This study for the first time documented that oribatid mites feed on certain ectomycorrhizal fungi.     

The effect of non-pathogenic phylloplane fungi on life-history traits of urticae (Acari: Tetranychidae)

The direct effect of three common non-pathogenic phylloplane fungi on the life-history traits of Tetranychus urticae Koch was investigated on intact leaves of the garden bean Phaseolus vulgaris L. (var. Tendergreen Improved), under conditions of low and high water-deficit stress. The survival rate of T. urticae was always reduced by an increase in water-deficit stress, whereas the effect of the fungi depended in part on the watering regime. On two of the three fungi tested, Alternaria alternata (Fr.) Keissler and Epicoccum nigrum Link, the mites showed a significant increase in net reproduction compared to those reared under control (i.e. no fungi added) conditions, independent of water-deficit stress, resulting in a higher intrinsic rate of increase. The third fungus tested, Cladosporium cladosporioides (Fres.) de Vries, had a similar response with respect to net reproduction when reared under low water-deficit stress but not when reared under a high water-deficit stress. There was, however, no evidence of an interactive effect with water-deficit stress and presence or absence of fungus affecting the net reproduction of the mites, for any of the three fungal species tested. An interactive effect between fungal species and water-deficit stress was observed for the intrinsic rate of increase of the mites. It is suspected that the fungi are acting as a source of an otherwise limiting resource and the differences observed between the fungal treatments is due to differences in consumption, based on the different sizes of their conidia.     

Endophyte communities vary in the needles of Norway spruce clones

Endophytic fungi show no symptoms of their presence but can influence the performance and vitality of host trees. The potential use of endophytes to indicate vitality has been previously realized, but a standard protocol has yet to be developed due to an incomplete understanding of the factors that regulate endophyte communities. Using a culture-free molecular approach, we examined the extent to which host genotype influences the abundance, species richness, and community composition of endophytic fungi in Norway spruce needles. Briefly, total DNA was extracted from the surface-sterilized needles of 30 clones grown in a nursery field and the copy number of the fungal internal transcribed spacer (ITS) region of ribosomal DNA was estimated by quantitative PCR. Fungal species richness and community composition were determined by denaturing gradient gel electrophoresis and DNA sequencing. We found that community structure and ITS copy number varied among spruce clones, whereas species richness did not. Host traits interacting with endophyte communities included needle surface area and the location of cuttings in the experimental area. Although Lophodermium piceae is considered the dominant needle endophyte of Norway spruce, we detected this species in only 33 % of samples. The most frequently observed fungus (66 %) was the potentially pathogenic Phoma herbarum. Interestingly, ITS copy number of endophytic fungi correlated negatively with the richness of ectomycorrhizal fungi and thus potential interactions between fungal communities and their influence on the host tree are discussed. Our results suggest that in addition to environmental factors, endophyte communities of spruce needles are determined by host tree identity and needle surface area.     

贵州省思南县不同生境区域鼠形动物体表革螨群落对比研究

1988年对贵州省思南县不同生境区域啮齿目及食虫目动物体表革螨群落的调查结果表明:室内生境革螨种类少、多样性低、优势种突出,以毒厉螨及柏氏禽刺螨为优势种;野外生境革螨种类丰富、多样性高、优势种不突出。其中海拔较高的山区和丘陵地带野外生境革螭群落为同一类型,以毒厉螨及土尔克历螨为优势种,低海拔平坝地带为另一类型,以鼠颚毛厉螨为优势种。     

Fungi Associated with Females and Cysts of Heterodera glycines in a Florida Soybean Field

Fungal colonization was determined for females and cysts of Heterodera glycines on soybean roots or in rhizosphere soil from a Florida soybean field. A total of 1,620 females and cysts were examined in 1991, and 1,303 were examined in 1992. More than 35 species of fungi were isolated from females and cysts. The frequency of fungi colonizing white and yellow females was low, but a high frequency of fungi was encountered in brown cysts, which increased with time of exposure of the cysts to the soil. No single fungal species predominated in the nematode females or cysts in this field. Rarely was a female or cyst colonized by more than one fungus. The common fungi isolated from the females and cysts were Neocosmospora vasinfecta, Fusarium solani, Fusarium oxysporum, Dictyochaeta coffeae, Dictyochaeta heteroderae, Pyrenochaeta terrestris, Exophiala pisciphila, Gliocladium catenulatum, Stagonospora heteroderae, and a black yeast-like fungus. The communities of common fungal species isolated from cysts in several regions in the southeastern United States appear to be similar.     

Rhizosphere Colonization and Control of Meloidogyne spp. by Nematode-trapping Fungi

The ability of nematode-trapping fungi to colonize the rhizosphere of crop plants has been suggested to be an important factor in biological control of root-infecting nematodes. In this study, rhizosphere colonization was evaluated for 38 isolates of nematode-trapping fungi representing 11 species. In an initial screen, Arthrobotrys dactyloides, A. superba, and Monacrosporium ellipsosporum were most frequently detected in the tomato rhizosphere. In subsequent pot experiments these fungi and the non-root colonizing M. geophyropagum were introduced to soil in a sodium alginate matrix, and further tested both for establishment in the tomato rhizosphere and suppression of root-knot nematodes. The knob-forming M. ellipsosporum showed a high capacity to colonize the rhizosphere both in the initial screen and the pot experiments, with more than twice as many fungal propagules in the rhizosphere as in the root-free soil. However, neither this fungus nor the other nematode-trapping fungi tested reduced nematode damage to tomato plants.     

云南省小兽体表革螨名录初报

1994年至2004年在云南省境内共调查了25个县（市）,共捕获53种啮齿目、食虫目、攀口目、兔形目和翼手目小兽10803只,从其体表分检出革螨68572只,隶属10科26属80种,其中云南新记录种12种;结合相关文献记载,到目前为止,整个云南省小兽体表革螨共有10科33属112种。