Accumulation of heat-shock proteins 70 in insect fat bodies as evidence of stress in Microsporidia-infected insects

At present a concept prevails that pathological alterations in insect hosts infected with microsporidia, and those associated with hormone imbalance may be explained by the production of juvenile hormone-like (JH) substances by microsporidia. According to another view point, this pathology is a consequence of the host response. We suggested that the microsporidian infection can provoke a stress reaction in insects, which may cause JH secretion by these insects. To confirm this hypothesis, we have analysed major stress protein Hsp70 levels in the infected insects. Using affinity chromatography on ATP-agarose and immunoblotting, we have shown that Hsp70 was accumulated in infected crickets, and that it was the host protein. The consequence of events accompanying the infection in the insects is discussed in relation to the response of hormonal system of the host organism.     

Impact of microsporidia on hormonal balance in insect hosts

Microsporidia (M) is a phylum of protists parasitizing obligatory in animal cells. Long way of adaptation of M to intracellular parasitism resulted in establishment of quite close relationships between the parasite and its host. Different species of M induce in their hosts symptoms similar to those caused by misbalance of juvenile hormone (JH) and ecdysone. M infection leads to pathology of different hormone-dependent functions such as cell differentiation and specialization, molting, metamorphosis, diapause and reproduction of insects. The signs of hormonal dysfunction evidence for elevated titer of JH in M-infected insects. Two possible explanation of this could be offered: JH secretion by M or specific influence of the parasites on the insect endocrine systems. Impact on insect endogenous JH titer by M could be mediated by affection of secretory activity of corpora allata or by suppression of enzymatic degradation of JH. According to different hypotheses, insect hormonal status during microsporidiosis could be modified by a) insect host stress-reaction, b) exhaustion of insect host reserves, characteristic for acute phase of the disease, c) destruction of infected insect cells and tissues during mass sporogenesis of M. Data found in literature and provided by our experiments evidence for presence of JH analogues or juvenilizing substance in the extracts of M spores. From detailed examination of pathological process it is also seen that juvenilizing effect of M infection is usually restricted to the invaded regions of tissues (i.e. expressed locally) but not a systemic one. Ability of M to modify morpho-functional features of infected tissues at the level of hormonal regulation is undoubtfully a prominent adaptation for stabilizing "microsporidia-insect" parasite-host systems.     

Cytological variation and pathogenicity of the bumble bee parasite Nosema bombi (Microspora, Nosematidae)

In three field seasons, 2003-2005, bumble bees were collected in southern Sweden and eastern Denmark in search of microsporidian parasites. Of the 16 bumble bee species studied, microsporidia were found in Bombus hortorum, Bombus hypnorum, Bombus lapidarius, Bombus lucorum, Bombus pascuorum, Bombus pratorum, Bombus ruderarius, Bombus subterraneus and Bombus terrestris. Only one microsporidian species, Nosema bombi, was recorded. A microsporidium found in B. pratorum differed cytologically from microsporidia of the other host species. In the most frequently infected host, B. terrestris, the prevalence was 20.6%. Totally 1049 specimens were dissected. The light microscopic and ultrastructural cytology and pathology of N. bombi is described with focus on the variation recorded. Variation was especially prominent in the shape, size and coupling of spores, and in the length and arrangement of the polar filament. In four host species microsporidian infection was restricted to peripheral fat cells.     

鳞翅目昆虫病原微孢子虫研究进展

微孢子虫广泛存在于鳞翅目昆虫中,是一类重要的病原微生物。微孢子虫病一方面影响野外昆虫种群的自然平衡,另一方面对家蚕、柞蚕等经济和资源昆虫造成了严重的危害。微孢子虫分子生物学研究基础相对薄弱,再加上微孢子虫表面坚厚的孢壁,无疑增加了研究难度。随着核酸、蛋白质等生物大分子分离制备方法和高通量测序技术的不断更新发展,基于各种组学(Omics)研究微孢子虫的工作方兴未艾,并且有了一些重要的发现。本文综述了微孢子虫与鳞翅目昆虫寄主的相互作用及寄生于鳞翅目昆虫的病原微孢子虫基因组、转录组和蛋白质组进展情况,以期为微孢子虫的深入研究提供参考。这些昆虫微生物研究将为鳞翅目害虫生物防治提供新的思路,并对家蚕等经济昆虫微粒子病的诊断、防控及治疗产生积极影响。     

Fish microsporidia: fine structural diversity and phylogeny

Structural diversity of fish microsporidian life cycle stages and of the host-parasite interface is reviewed. In the infected cell of the fish host, microsporidia may either cause serious degradation of the cytoplasm and demise of the cell, or they may elicit host cell hypertrophy, producing a parasite-hypertrophic host cell complex, the xenoma. The structure of the xenoma and of its cell wall may differ according to the genus of the parasite, and seems to express properties of the parasite rather than those of the host. In merogony, the parasite cell surface interacts with the host cell in diverse ways, the most conspicuous being the production of thick envelopes of different types. Sporogony stages reveal different types of walls or membranes encasing the sporoblasts and later the spores and these envelopes may be of host or parasite origin. Nucleospora differs from all other fish microsporidia by its unique process of sporogony. Except for the formation of conspicuous xenomas, there are no essentially different structures in fish-infecting microsporidia compared with microsporidia from other hosts. Although the structures associated with the development of fish microsporidia cannot be attributed importance in tracing the phylogeny, they are relevant for practical determination and assessing the relation to the host. The possibility of the existence of an intermediate host is discussed. Higher-level classification of Microsporidia is briefly discussed and structure and evolutionary rates in microsporidian rDNA are reviewed. Discussion of rDNA molecular phylogeny of fish-infecting microsporidia is followed by classification of these parasites. Most form a rather cohesive clade. Outside this clade is the genus Nucleospora, separated at least at the level of Order. Within the main clade, however, there are six species infecting hosts other than fish. Based on data available for analysis, a tentative classification of fish-infecting microsporidia into five groups is proposed. Morphologically defined groups represent families, others are referred to as clades. Group 1, represented by family Pleistophoridae, includes Pleistophora, Ovipleistophora and Heterosporis; Vavraia and Trachipleistophora infect non-fish hosts. Group 2, represented by family Glugeidae, is restricted to genus Glugea and Tuzetia weidneri from crustaceans. Group 3 comprises three clades: Loma and a hyperparasitic microsporidian from a myxosporean; Ichthyosporidium and Pseudoloma clade and the Loma acerinae clade. For the latter species a new genus has to be established. Group 4 contains two families, Spragueidae with the genus Spraguea and Tetramicridae with genera Microgemma and Tetramicra, and the Kabatana and Microsporidium seriolae clade. Group 5 is represented by the family Enterocytozoonidae with the genus Nucleospora and mammal-infecting genus Enterocytozoon.     

Wing dimorphism and the migratory syndrome: Correlated traits for migratory tendency in wing dimorphic insects

The hypothesis that the morphological, physiological, and behavioral traits comprising the migratory syndrome in insects are genetically correlated through pleiotropic effects of genes controlling the titre of a common hormonal determinant is explored. Evidence that juvenile hormone (JH) influences the component traits of the migratory syndrome is presented, and thus JH is assumed to be the underlying, common determinant. However, readers are cautioned that this does not imply that JH is solely responsible for these traits, nor is this necessary for the arguments presented. For wing dimorphic taxa, the “correlated traits hypothesis” predicts covariance within wing morphs between JH titre and the proportion winged. Four simple genetic models for wing-morph determination are considered: single-locus with short-winged (SW) dominant; single-locus with long-winged (LW) dominant; polygenic, fixed threshold, shifting distribution; and polygenic, shifting threshold, fixed distribution. In each case, wing morphology is assumed to be a threshold trait with the liability being JH titre at some critical stage of development. All models predict covariation between %LW and the mean JH titre of at least one of the wing morphs, but the form and direction of the relationship depends critically on the genetic model used. The results suggest that we should expect the traits associated with the migratory syndrome, and hence the trade-offs associated with the evolution of wing dimorphism, to be correlated with proportion winged and, in this sense, to be frequency-dependent.     

Some old concepts and new findings on hormonal control of insect morphogenesis

By means of the artificially induced heterochronic developmental deviations represented by local prothetelies and metathetelies it has been possible to investigate the individual developmental fates of ontogenetically different tissues, such as larval, pupal, and adult epidermal cells, in one and the same body and under the identical concentration of juvenile hormone (JH) in the haemolymph.In contrast to the widely accepted hormonal theories which claim that the kind of morphogenesis is determined by large, intermediate, and low titres of JH, the heterochronic character of the tissues never developed into a uniform population of homomorphic epidermal cells. Instead, in the presence of effective amounts of JH the heterochronic pattern has been fully preserved and carried on into the next developmental instar. Moreover, in the absence of the effective JH amounts the ontogenetically different tissues, such as larval and pupal epidermal cells, simultaneously undergo their respective morphogenetic developments, i.e. larval-pupal and pupal-adult morphogenesis in the same hormonal milieu. It is concluded that the selective factor in determination of the kind of morphogenetical changes is not an altered JH titre but the extant, previously attained degree of ontogenetic structural differentiation. It has been demonstrated that JH can temporarily and reversibly inhibit the morphogenetic progress at quite different ontogenetic levels but it cannot cause a ‘reversal of metamorphosis’ at any of these levels.Under specific experimental conditions the larval epidermal cells can undergo pupal and adult morphogenesis without secreting the pupal cuticle. However, the pupal morphogenetic interstage, whether with the cuticle or without the pupal cuticle, constitutes an obligatory developmental step. Further, it appears that an absence of JH may represent an important condition but not a real cause of insect metamorphosis, as presumed in some other hormonal concepts. Thus, chromosomal duplications or cellular divisions in the absence of JH have not committed the cells to morphogenesis unless provided by an additional stimulus of endogenous prothoracic gland hormone or exogenous ecdysterone. An important factor in understanding the hormonal control of insect morphogenesis is the critical timing of the respective morphogenetic steps. This corresponds closely with the duration of the pharate phases in insect development. Possible hormonal mechanisms concerned in the regulation of morphogenesis in endopterygote insects have been outlined.     

Coexistence of three microsporidia parasites in populations of the freshwater amphipod Gammarus roeseli: evidence for vertical transmission and positive effect on reproduction

We investigated the prevalence, transmission mode and fitness effects of infections by obligatory intracellular, microsporidian parasites in the freshwater amphipod Gammarus roeseli. We found three different microsporidia species in this host, all using transovarial (vertical) transmission. All three coexist at different prevalences in two host populations, but bi-infected individuals were rarely found, suggesting no (or very little) horizontal transmission. It is predicted that vertically-transmitted parasites may exhibit sex-specific virulence in their hosts, or they may have either positive or neutral effects on host fitness. All three species differed in their transmission efficiency and infection intensity and our data suggest that these microsporidia exert sex-specific virulence by feminising male hosts. The patterns of infection we found exhibit convergent evolution with those of another amphipod host, Gammarus duebeni. Interestingly, we found that infected females breed earlier in the reproductive season than uninfected females. This is the first study, to our knowledge, to report a positive effect of microsporidian infection on female host reproduction.     

Natural distribution and co-infection patterns of microsporidia parasites in the Daphnia longispina complex

Microsporidia are intracellular parasites, frequently infecting the planktonic crustacean Daphnia. Questioning the ability to detect and identify microsporidia with conventional microscopic techniques, we applied molecular methods in order to investigate the distribution and co-infection patterns of this parasite among 8 communities of the Daphnia longispina hybrid complex. Eight microsporidian taxa were detected, including 3 that previously had not been characterized genetically. Microsporidian communities from nearby lakes were found to be more similar to each other, apparently due to short distance dispersal via secondary hosts. Moreover, we detected seasonal (but not interannual) changes in microsporidian community structure. With some microsporidia being host-specific, these changes might have resulted from seasonal changes in host taxon and clonal composition. The 2 dominant and closely related parasite species were found mainly in single infections, whereas another pair of related microsporidians was found predominantly in co-infections; suggesting species-level differences in the ability to colonize infected hosts. By applying molecular methods, we were not only able to unambiguously identify parasite taxa but also to reveal multiple infections that otherwise would have remained undetected. Given the increased level of accuracy and sensitivity, we highly recommend molecular approaches in future parasite surveys of Daphnia infections.     

Luna stain, an improved selective stain for detection of microsporidian spores in histologic sections

Microsporidia in histologic sections are most often diagnosed by observing spores in host tissues. Spores are easy to identify if they occur in large aggregates or xenomas when sections are stained with hematoxylin and eosin (H&E). However, individual spores are not frequently detected in host tissues with conventional H&E staining, particularly if spores are scattered within the tissues, areas of inflammation, or small spores in nuclei (i.e. Nucleospora salmonis). Hence, a variety of selective stains that enhance visualization of spores is recommended. We discovered that the Luna stain, used to highlight eosinophils, red blood cells, and chitin in arthropods and other invertebrates, also stains spores of Pseudoloma neurophilia. We compared this stain to the Gram, Fite's acid fast, Giemsa, and H&E stains on 8 aquatic microsporidian organisms that were readily available in our 2 laboratories: Loma salmonae, Glugea anomala, Pseudoloma neurophilia, Pleistophora hyphessobryconis, Pleistophora vermiformis, Glugea sp., Steinhausia mytilovum, and an unidentified microsporidian from UK mitten crabs Eriocheir sinensis. Based on tinctorial properties and background staining, the Luna stain performed better for detection of 6 of the 8 microsporidia. Gram stain was superior for the 2 microsporidia from invertebrates: S. mytilovum and the unidentified microsporidian from E. sinensis.     

Long-Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Thirty-one species of microsporidia, isolated from insects and stored in liquid nitrogen for up to 25 yr, were infectious when removed from liquid nitrogen. The natural hosts of all of these microsporidia were terrestrial insects, representing six different insect orders: Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera, and Orthoptera. All microsporidia from terrestrial insects that were tested survived storage in liquid nitrogen, while Nosema algerae , a microsporidium from aquatic mosquito hosts did not survive freezing in liquid nitrogen. A Nosema species from the alfalfa weevil, Hypera postica , lost some infectivity in a water storage medium after 25 yr in liquid nitrogen. Liquid nitrogen storage of microsporidian spores in 50% and 100% glycerol media reduced loss of infectivity and is recommended for extended storage of microsporidia from terrestrial insect hosts.     

Effects of microsporjdia on the striated parietal muscle of Rhynchosciara angelae (Diptera: Sciaridae)

Nuclear hypertrophy and RNA synthesis were studied in the multinucleated striated muscle of Rhynchosciara angelae infected by the microsporidia Thelohania sp. Infection varied from fibers with few microsporidian cells to fibers completely occupied by the infective agent. Usually, the terminal portions of the fibers are free of microsporidia. The host-cell nuclei located in these regions (OM nuclei) are less affected than the nuclei of the same fiber that are completely surrounded by the cells of the parasite (IM). There is a correlation between the volume attained by the OM nuclei and their location in the fiber: the closer the nuclei to the microsporidia the higher the nuclear hypertrophy. Despite the errors peculiar to the autoradiography, grain counting showed that the nuclei of infected fibers are more active in the incorporation of RNA precursor than the normal noninfected fibers. Two additive mechanisms are suggested through which the microsporidia could influence the host-cell nucleus: (1) enhancement of the metabolism induced by juvenile hormone and (2) a further increase in the overall metabolism to repair the disruption imposed by the microsporidian development.     

The Interplay between Patency,Microsomal Na(+)/K(+) ATPase Activity and Juvenile Hormone,in Tenebrio molitor Parasitized by Hymenolepsis diminuta

Beetles infected with metacestodes of the rat tapeworm, Hymenolepis diminuta, exhibit reduced fecundity, due to alterations in vitellogenesis. Follicle cell patency is retarded and inefficient vitellogenin uptake ensues. Here, we have reassessed patency and its stimulation by JH III at day 3 post-infection, when the most detrimental changes are observed in other ovarian processes. In Rhodnius prolixus, patency is believed to be brought about by the action of a JH-dependent membrane-bound Na(+)/K(+) ATPase (EC 3.6.1.3); however, this had not been established in Tenebrio molitor. Therefore, the properties of the enzyme, with respect to optimal assay conditions and juvenile hormone dependency, are reported. Maximal stimulation occurred between 50 and 500 nM JH III, a range over which greatest increases in patency were also observed. In infected insects, a 35% reduction in Na(+)/K(+) ATPase activity was noted, but exposure to 50 nM JH III is sufficient for stimulation to a specific activity 89% that of JH-treated controls. In a similar fashion, patency in infected insects is reduced, but can be 'rescued' by 50 nM JH III. Moreover, in the absence of exogenous hormone, patency in infected beetles can be elevated to control levels after in vitro culture (6 h), with exchange of medium every 2 h. The possibility that such reversible decreases in enzyme activity and patency are caused by a JH binding inhibitor molecule is discussed. Copyright 1997 Elsevier Science Ltd. All rights reserved     

The Genome of Spraguea lophii and the Basis of Host-Microsporidian Interactions

Microsporidia are obligate intracellular parasites with the smallest known eukaryotic genomes. Although they are increasingly recognized as economically and medically important parasites, the molecular basis of microsporidian pathogenicity is almost completely unknown and no genetic manipulation system is currently available. The fish-infecting microsporidian Spraguea lophii shows one of the most striking host cell manipulations known for these parasites, converting host nervous tissue into swollen spore factories known as xenomas. In order to investigate the basis of these interactions between microsporidian and host, we sequenced and analyzed the S. lophii genome. Although, like other microsporidia, S. lophii has lost many of the protein families typical of model eukaryotes, we identified a number of gene family expansions including a family of leucine-rich repeat proteins that may represent pathogenicity factors. Building on our comparative genomic analyses, we exploited the large numbers of spores that can be obtained from xenomas to identify potential effector proteins experimentally. We used complex-mix proteomics to identify proteins released by the parasite upon germination, resulting in the first experimental isolation of putative secreted effector proteins in a microsporidian. Many of these proteins are not related to characterized pathogenicity factors or indeed any other sequences from outside the Microsporidia. However, two of the secreted proteins are members of a family of RICIN B-lectin-like proteins broadly conserved across the phylum. These proteins form syntenic clusters arising from tandem duplications in several microsporidian genomes and may represent a novel family of conserved effector proteins. These computational and experimental analyses establish S. lophii as an attractive model system for understanding the evolution of host-parasite interactions in microsporidia and suggest an important role for lineage-specific innovations and fast evolving proteins in the evolution of the parasitic microsporidian lifecycle.     

A microsporidium,Nosema pilicornis sp. n., of the purslane sawfly,Schizocerella pilicornis

A new microsporidian species, Nosema pilicornis, which infects the purslane sawfly, Schizocerella pilicornis, is described. This microsporidium infects most body tissues of the host. N. pilicornis was compared to other microsporidian species infecting Hymenoptera and to a group of similar microsporidia infecting Lepidoptera. N. pilicornis could be distinguished from all other microsporidian species on the basis of host range and ultrastructural characteristics of the spore. Spores were oval, containing 11 to 12 polar filament coils, and the polar filament had an angle of tilt of about 80°. N. pilicornis infected lepidopteran larvae, but only when heavy spore dosages were fed to early larval instars. S. pilicornis is a good but sporadic biological control agent of common purslane, Portulaca oleracea, a pernicious weed of vegetable, ornamental, and orchard crops. N. pilicornis, which is transovarially transmitted and causes high mortality in infected larvae, affects the performance of S. pilicornis as a biological control agent.     

Microsporidia Are Natural Intracellular Parasites of the Nematode Caenorhabditis elegans

For decades the soil nematode Caenorhabditis elegans has been an important model system for biology, but little is known about its natural ecology. Recently, C. elegans has become the focus of studies of innate immunity and several pathogens have been shown to cause lethal intestinal infections in C. elegans. However none of these pathogens has been shown to invade nematode intestinal cells, and no pathogen has been isolated from wild-caught C. elegans. Here we describe an intracellular pathogen isolated from wild-caught C. elegans that we show is a new species of microsporidia. Microsporidia comprise a large class of eukaryotic intracellular parasites that are medically and agriculturally important, but poorly understood. We show that microsporidian infection of the C. elegans intestine proceeds through distinct stages and is transmitted horizontally. Disruption of a conserved cytoskeletal structure in the intestine called the terminal web correlates with the release of microsporidian spores from infected cells, and appears to be part of a novel mechanism by which intracellular pathogens exit from infected cells. Unlike in bacterial intestinal infections, the p38 MAPK and insulin/insulin-like growth factor (IGF) signaling pathways do not appear to play substantial roles in resistance to microsporidian infection in C. elegans. We found microsporidia in multiple wild-caught isolates of Caenorhabditis nematodes from diverse geographic locations. These results indicate that microsporidia are common parasites of C. elegans in the wild. In addition, the interaction between C. elegans and its natural microsporidian parasites provides a system in which to dissect intracellular intestinal infection in vivo and insight into the diversity of pathogenic mechanisms used by intracellular microbes.     

The effects of two microsporidian pathogens on the two-spotted lady beetle, Adalia bipunctata L. (Coleoptera: Coccinellidae)

Two-spotted lady beetles, Adalia bipunctata L. are available for biological pest control in North America. Lady beetles are known to host microsporidia and although these pathogens are able to infect more than one host under laboratory conditions, little is known regarding the effects of more than one microsporidian pathogen on host fitness. In this study, egg cannibalism was used to examine the effects of the microsporidium Tubulinosema hippodamiae from Hippodamia convergens and an undescribed microsporidium from A. bipunctata (alone and in combination) on A. bipunctata host fitness (larval development and mortality, sex ratio, adult fecundity and longevity). Development was prolonged significantly for larvae that were infected with the undescribed microsporidium but T. hippodamiae had no effect and as a result, conclusions could not be made regarding the effects of both pathogens on larval development. The two microsporidia had no effect on sex ratios (1♀:1♂) or on adult fecundity and longevity. Spores were detected in the majority of smear preparations of individuals that were fed microsporidia-infected eggs and molecular analysis confirmed the identity of both pathogens in sampled individuals. T. hippodamiae spores were smaller than spores of the undescribed microsporidium (3.76±0.03×2.32±0.02 μm and 5.43±0.06×2.75±0.03 μm, respectively) and although the former stained less intensely than did those of latter, spores of the two pathogens are difficult to differentiate when examined by light microscopy alone. The ability of some microsporidia to infect more than one lady beetle host makes it difficult to conclude with certainty as to the number of species that are present in infected Adalia when specimens are examined solely by light microscopy.     

Interplay of JH, 20E and biogenic amines under normal and stress conditions and its effect on reproduction

Juvenile hormone (JH) and 20-hydroxyecdysone (20E) are well known to play a gonadotropic role in adult insects. In Drosophila the mechanism of reciprocal regulation of JH and 20E is shown to be responsible for their proper balance. Dopamine is a mediator in this JH and 20E interplay. A proper balance between JH and 20E is crucial for the normal progress of oogenesis. An imbalance of gonadotropins leads to reproductive defects: a rise in JH titre leads to oviposition arrest, a rise in 20E level, to the degradation of vitellogenic oocytes. Upon a change in the level of one of the gonadotropins, the balance is restored owing to the relative change in the titre of the other.     

Microsporidiosis in the wax moth Galleria mellonella (Lepidoptera: Pyralidae) caused by Vairimorpha ephestiae (Microsporidia: Burenellidae)

An experimental microsporidiosis of the wax moth caterpillars from laboratory population had been caused by oral infecting of early stages larvae and by intracavity injections of the spores of the microsporidian species Vairimorpha ephestiae. Peculiarities of microsporidiosis proceeding, manifestations of host defence reactions, and also an effect of the temperature of caterpillars cultivation and conditions of spores keeping on liability of the insects to the infection were studied. The effect of the microsporidia on the host organism was the early death or the delay of larvae development, but in several cases external manifestations of the effect of the parasite on the host were absent. The development of the parasites from the moment of infecting to the appearing of the mature spores congestions in the host organism proceeded 6 days. Microsporidia invaded insect fat body and caused its hypertrophy and disappearance of lipid granules. In the intestine and salivary glands microsporidia were not observed in the period from 6 to 16 day of the development. On the final stage of microsporidiosis the all contents of fatty tissue cells were replaced by spores of microsporidia. Under microscope only diplocaryotic spores of the Nozema type had been found in infected and died specimens, but not octospores. The spores threw out polar tubes under the change of pH in incubating solution from neutral to alkaline. The effects of microsporidiosis on the wax moth haemolymph were the increased rate of prohaemocytes, appearing of multinuclear free-circulating cells at 6 day after infection, and suppression of the reaction of haemolymph melanization with the mass sporogenesis of the parasite. The characteristic symptom of the wax moth microsporidiosis had been revealed, accumulation of black points and small spots of irregular form under cuticle ("reaction of attretization"). Increase of the temperature of insect cultivation up to 32 degrees C during 3 days after infection contributed to the full deliverance of the insects from the infection in first and second generations. It can be considered as a method of treatment of wax moth laboratory colonies from microsporidiosis. Oral infection of III and IV stage caterpillars by the spores being kept during 3-6 months under 4 degrees C in form of water suspension caused the death of 63.0-61.5 and 91% of caterpillars being cultivated under 25 and 21 degrees C respectively. Under the temperature of cultivation equal 30 degrees C the mortality did not differ from the control sample (8-10%). The spores extracted from dried bodies of caterpillars lost their vitality. It was demonstrated by the test on infectious ability in vivo and by acridine orange staining. This host-parasite system appears to be perspective in investigations of resistance mechanisms in insects and immunosuppressive features of entomopathogen microsporidia.