Microsporidian Species Research Articles

Molecular detection and genotyping of microsporidia species in chickens in Turkey

Microsporidia are obligate intracellular pathogens that infect various hosts including invertebrates and vertebrates. Despite the importance, knowledge on the prevalence and molecular characteristics of microsporidia in chickens is limited, and no data are available for Turkey. A total of 300 fecal samples from chickens in the Central Anatolia Region of Turkey were analyzed by using a nested polymerase chain reaction assay targeting the rRNA internal transcribed spacer (ITS) region for the common microsporidia species. Corresponding PCR amplicons from the positive samples were sequenced for genotyping. Enterocytozoon bieneusi was identified in 22 (7.3 %) samples, whereas Encephalitozoon spp. was not detected. The prevalence of E. bieneusi was 63.6 % in Kayseri and 36.4 % in Nevsehir provinces, and 8.8 % in soft fecal samples and 9.7 % in diarrhoeic samples. No infections were found in Kirsehir Province. Significant differences were found for the distribution of E. bieneusi among provinces and fecal conditions. Infections were found only in free-range chickens. As a result of ITS region sequencing, two genotypes were characterized. The novel genotype ERUNT1 (n = 21), belonging to zoonotic group 1, was the most common genotype throughout the study area. The other known genotype, ERUSS1 (n = 1), had a restricted distribution and was previously detected in cattle and sheep in the same region. Our study provides the first data on microsporidia species from chickens in Turkey. None of these genotypes have been reported in humans; thus, the risk potential for public health is limited but needs further investigation.

A new method of metabarcoding Microsporidia and their hosts reveals high levels of microsporidian infections in mosquitoes (Culicidae).

DNA metabarcoding offers new perspectives, especially with regard to the high‐throughput identification and diagnostics of pathogens. Microsporidia are an example of widely distributed, opportunistic and pathogenic microorganisms in which molecular identification is important for both environmental research and clinical diagnostics. We have developed a method for parallel detection of both microsporidian infection and the host species. We designed new primer sets: one specific for the classical Microsporidia (targeting the hypervariable V5 region of small subunit [ssu] rDNA), and a second one targeting a shortened fragment of the COI gene (standard metazoan DNA‐barcode); both markers are well suited for next generation sequencing. Analysis of the ssu rDNA data set representing 607 microsporidian species (120 genera) indicated that the V5 region enables identification of >98% species in the data set (596/607). To test the method, we used microsporidians that infect mosquitoes in natural populations. Using mini‐COI data, all field‐collected mosquitoes were unambiguously assigned to seven species; among them almost 60% of specimens were positive for at least 11 different microsporidian species, including a new microsporidian ssu rDNA sequence (Microsporidium sp. PL01). Phylogenetic analysis showed that this species belongs to one of the two main clades in the Terresporidia. We found a high rate of microsporidian co‐infections (9.4%). The numbers of sequence reads for the operational taxonomic units suggest that the occurrence of Nosema spp. in co‐infections could benefit them; however, this observation should be retested using a more intensive host sampling. Our results show that DNA barcoding is a rapid and cost‐effective method for deciphering sample diversity in greater resolution, including the hidden biodiversity that may be overlooked using classical methodology.

Multilocus Sequence Typing of Enterocytozoon bieneusi Isolates From Various Mammal and Bird Species and Assessment of Population Structure and Substructure.

Enterocytozoon bieneusi is one of the most common intestinal pathogens in humans and animals. E. bieneusi has been confirmed to be complex microsporidian species. Approximately 500 ITS genotypes of E. bieneusi have been defined. With the establishment and application of multilocus sequencing typing and population genetic tools in E. bieneusi, the studies on these aspects have been carried out worldwide, but little information is available. To understand genetic variation of mini-/micro-satellites and the population structure and substructure of E. bieneusi in northeastern China, 305 E. bieneusi DNA specimens composed of 28 ITS genotypes were from 13 mammal species and five bird species in the investigated areas. They were characterized by nested-PCR amplification and sequencing at four mini-/micro-satellite loci (MS1, MS3, MS4, and MS7). At the MS1, MS3, MS4, and MS7 loci, 153 (50.16%), 131 (42.95%), 133 (43.61%), and 128 (41.97%) DNA specimens were amplified and sequenced successfully with 44, 17, 26, and 24 genotypes being identified, respectively. Multilocus genotypes (MLGs) showed a higher genetic diversity than ITS genotypes. 48 MLGs were produced out of 90 ITS-positive DNA specimens based on concatenated sequences of all the five genetic loci including ITS. Linkage disequilibrium (LD) and limited genetic recombination were observed by measuring LD using both multilocus sequences and allelic profile data, indicating an overall clonal population structure of E. bieneusi in the investigated areas. These data will aid in the longitudinal tracking of the attribution of source of infection/contamination and in elucidating transmission dynamics, and will provide valuable information for making efficient control strategies to intervene with and prevent occurrence of microsporidiosis caused by E. bieneusi among animals and transmission of E. bieneusi from animals to humans in the investigated areas. Phylogenetic and network analyses identified three different subgroups, revealing the presence of host-shaped segregation and the absence of geographical segregation in E. bieneusi population. Meanwhile, the MLGs from zoonotic ITS genotypes were observed to be basically separated from the MLGs from host-adapted ones. Assessment of substructure will have a reference effect on understanding of zoonotic or interspecies transmission of E. bieneusi and evolution direction from zoonotic genotypes to host-adapted genotypes.

Zoonotic potential of Enterocytozoon bieneusi in pre-weaned Korean native calves

BackgroundEnterocytozoon bieneusi is the most common microsporidian species infecting humans and various animals worldwide. To date, there has been limited information on the prevalence of infection and genotypes of E. bieneusi in cattle in the Republic of Korea. Therefore, this study investigated the prevalence and genotypes of E. bieneusi circulating in pre-weaned Korean native calves and determined the age pattern of E. bieneusi infection and the relationship between E. bieneusi infection and diarrhea.MethodsThe prevalence of E. bieneusi infection in pre-weaned Korean native calves was screened by polymerase chain reaction. PCR-positive products were sequenced to determine the genotype of E. bieneusi. A Chi-square analysis was used to compare the association between diarrhea and the infection rate of E. bieneusi in each age range or for all ages.ResultsPCR and sequencing analysis revealed an overall prevalence (16.9%, 53/314) of E. bieneusi in pre-weaned calves. The prevalence of E. bieneusi was highest in September (36.2%), followed by March (28.3%). Enterocytozoon bieneusi infection was associated with diarrhea in calves (χ2 = 5.82, P = 0.016). Our results also indicated that E. bieneusi infection was significantly associated with calf age (χ2 = 11.61, P = 0.003), and the prevalence of E. bieneusi infection was significantly higher in calves aged 21–40 days-old (odds ratio: 2.90, 95% confidence interval: 1.54–5.45; P = 0.001) than in those aged 1–20 days-old. Interestingly, the association between E. bieneusi infection and diarrhea was observed only in calves aged 1–20 days-old (χ2 = 5.82, P = 0.010). We identified three known genotypes, BEB4 (n = 12), BEB8 (n = 21) and J (n = 16), and three novel genotypes, BEB8-like (n = 21), KCALF1 (n = 1) and KCALF2 (n = 1). The genotype BEB8 was the most prevalent among all age groups. All genotypes identified in this study exhibited zoonotic potential.ConclusionsTo our knowledge, this is the first report of the genotype BEB4 in pre-weaned Korean native calves. Zoonotic E. bieneusi infection was prevalent in pre-weaned calves, indicating that cattle may play an important role as a reservoir host for E. bieneusi transmission to humans.

Pneumocystis jirovecii and microsporidia: An unusual coinfection in HIV patients?

Pneumocystis jirovecii and microsporidia species are recognized as opportunistic infectious pathogens in AIDS patients. Coinfection of both in one patient has been rarely reported. The aim of the present study was to investigate the coinfection of P. jirovecii and microsporidia in different tissues from AIDS deceased patients. Post mortem histological finding of P. jirovecii and microsporidia was demonstrated by means of the Grocott's methenamine silver and Brown Brenn staining, respectively. Molecular technique was used for identification and characterization of both fungi. Out of the 514 autopsied cases P. jirovecii and microsporidia species were identified in 53 (10.3%) and 62 (12.1%) cases respectively. A total of five cases (0.97%) coinfected with Pneumocystis and microsporidia were recovered from all analyzed autopsies. Coinfection of Pneumocystis and microsporidia is very challenging and raises interesting issues about host-parasite relationship. The early diagnosis of both pathogens must be crucial to establish correct and early treatments, improve the patient's evolution, reducing the risk of death.

Characterization of Hsp70 gene family provides insight into its functions related to microsporidian proliferation

Heat shock protein 70 (Hsp70), a highly conserved protein family, is widely distributed in organisms and plays fundamental roles in biotic and abiotic stress responses. However, reports on Hsp70 genes are scarce in microsporidia, a very large group of obligate intracellular parasites that can infect nearly all animals, including humans. In this study, we identified 37 Hsp70 proteins from eight microsporidian genomes and classified them into four subfamilies (A–D). The number of Hsp70 genes in these microsporidia was significantly fewer than in Rozella allomycis and yeast. All microsporidian species contained genes from each subfamily and similar subcellular locations (mitochondria, endoplasmic reticulum, cytosol, and cytosol and/or nucleus), indicating that each Hsp70 member may fulfil distinct functions. The conserved structures and motifs of the Hsp70 proteins in the same subfamily were highly similar. Expression analysis indicated that the subfamily C cytosol (cyto)-associated Hsp70s is functional during microsporidia development. Immunofluorescence assays revealed that Cyto-NbHsp70 was cytoplasmically located in the proliferation-stage of Nosema bombycis. Cyto-NbHsp70 antiserum also labeled Encephalitozoon hellem within infected cells, suggesting that this antiserum is a potential molecular marker for labeling the proliferative phases of different microsporidian species. The propagation of N. bombycis was significantly inhibited following RNAi of Cyto-NbHsp70, indicating that Cyto-NbHsp70 is important for pathogen proliferation. Our phylogenetic data suggest that Hsp70 proteins evolved during microsporidia adaption to intracellular parasitism, and they play important roles in pathogen development.

Prevalence and genotype distribution of Enterocytozoon bieneusi in farmed raccoon dogs (Nyctereutes procyonoides) in Shandong Province, eastern China.

Enterocytozoon bieneusi is a common microsporidian species, which can infect humans and various species of animals. However, little is known about E. bieneusi prevalence and genotypes in farmed raccoon dogs (Nyctereutes procyonoides) in Shandong Province, China. In this study, a total of 356 fecal samples were collected from farmed raccoon dogs in Weihai, Weifang, and Yantai cities in Shandong Province, China. A total of 23 (6.5%) samples were E. bieneusi-positive by nested PCR amplification of the internal transcribed spacer (ITS) region of ribosomal DNA. Statistical analysis showed that E. bieneusi prevalence in male raccoon dogs was higher than that in female raccoon dogs, and the highest E. bieneusi prevalence was detected in adult raccoon dogs. Sequence analysis revealed four known E. bieneusi genotypes (D, type IV, CHG1, and Peru8), and type IV (11/23) was the predominant genotype. The genotypes type IV, Peru8, and CHG1 were reported in raccoon dogs for the first time in China. Phylogenetic analysis showed that three human-pathogenic genotypes (D, type IV, and Peru8) were clustered into group 1, and the CHG1 belonged to group 2. These findings expand the current understanding of E. bieneusi prevalence and genotype distribution in raccoon dogs in China. Our study also shows that raccoon dogs are hosts for E. bieneusi belonging to several genotypes, including zoonotic ones, highlighting the possibility of transmission of this pathogen between raccoon dogs and humans.

The Effects of Quantum Dots Nanoparticles on Caenorhabditis elegans infected with Microsporidia

Abstract Caenorhabditis elegans are model organisms that have intestinal lining similar to that of humans. They are transparent and can be easily cultivated in the lab. Nematocida parisii is an intracellular parasitic species of microsporidia that infect the intestines of C. elegans. Similar species cause microsporidiosis in humans. Microsporidiosis can cause severe digestive system symptoms including life-threatening diarrhea in immunocompromised as well as in immunocompetent individuals. Currently available treatments are not always sufficient and can be toxic. In this experiment, the effects of different nanoparticles (NPs) on the intestinal lining of C. elegans and N. parisii within are tested. It was reported that NPs can diffuse through cellular membranes and cause oxidative stress, DNA damage, and macrophage activation. Some NPs showed anti-bacterial and anti-fungal effects. If the treated C. elegans have a longer lifespan and a larger population density than untreated ones, then NPs can preferentially diffuse through the epithelial cells of the intestines and affect the infection. The number of thrashes of the C. elegans was used as a measure of their response to stimuli. Irritating and disrupting stimuli cause an initial increase in the movements but it is then followed with a sharp drop and the organisms may eventually die. The imaging of C. elegans was done using confocal microscopy with DIC. The preliminary results showed that the quantum dots of 655 nm emission wavelength and at a concentration of 0.8μM showed no significant difference that indicated a harmful effect on the C. elegans. Thus, different concentrations and longer exposure times will be implemented to compare their effects on the microsporidia.

The first case of microsporidiosis in Paramecium.

A new microsporidian species, Globosporidium paramecii gen. nov., sp. nov., from Paramecium primaurelia is described on the basis of morphology, fine structure, and SSU rRNA gene sequence. This is the first case of microsporidiosis in Paramecium reported so far. All observed stages of the life cycle are monokaryotic. The parasites develop in the cytoplasm, at least some part of the population in endoplasmic reticulum and its derivates. Meronts divide by binary fission. Sporogonial plasmodium divides by rosette-like budding. Early sporoblasts demonstrate a well-developed exospore forming blister-like structures. Spores with distinctive spherical shape are dimorphic in size (3.7 ± 0.2 and 1.9 ± 0.2 μm). Both types of spores are characterized by a thin endospore, a short isofilar polar tube making one incomplete coil, a bipartite polaroplast, and a large posterior vacuole. Experimental infection was successful for 5 of 10 tested strains of the Paramecium aurelia species complex. All susceptible strains belong to closely related P. primaurelia and P. pentaurelia species. Phylogenetic analysis placed the new species in the Clade 4 of Microsporidia and revealed its close relationship to Euplotespora binucleata (a microsporidium from the ciliate Euplotes woodruffi), to Helmichia lacustris and Mrazekia macrocyclopis, microsporidia from aquatic invertebrates.

The Presence and Distribution of Nosemosis Disease in Turkey

Nosemosis is one of the most important bee diseases causing economic losses in beekeeping, which is one of the significant reasons for Colony Collapse Disorder (CCD) in the world. Nosema apis and Nosema ceranae, the microsporidian species, are the main causative agents of Nosemosis in honey bees worldwide. This disease causes digestive system disorders, a decrease in the average life of bees and colony losses. In this review, the general characteristics of Nosemosis disease, and information about the situation in Turkey are given.

Cell invasion by intracellular parasites - the many roads to infection.

Intracellular parasites from the genera Toxoplasma, Plasmodium, Trypanosoma, Leishmania and from the phylum Microsporidia are, respectively, the causative agents of toxoplasmosis, malaria, Chagas disease, leishmaniasis and microsporidiosis, illnesses that kill millions of people around the globe. Crossing the host cell plasma membrane (PM) is an obstacle these parasites must overcome to establish themselves intracellularly and so cause diseases. The mechanisms of cell invasion are quite diverse and include (1) formation of moving junctions that drive parasites into host cells, as for the protozoans Toxoplasma gondii and Plasmodium spp., (2) subversion of endocytic pathways used by the host cell to repair PM, as for Trypanosoma cruzi and Leishmania, (3) induction of phagocytosis as for Leishmania or (4) endocytosis of parasites induced by specialized structures, such as the polar tubes present in microsporidian species. Understanding the early steps of cell entry is essential for the development of vaccines and drugs for the prevention or treatment of these diseases, and thus enormous research efforts have been made to unveil their underlying biological mechanisms. This Review will focus on these mechanisms and the factors involved, with an emphasis on the recent insights into the cell biology of invasion by these pathogens.

Updates to the sporadic knowledge on microsporidian infections in groundwater amphipods (Crustacea, Amphipoda, Niphargidae)

A set of 69 specimens from 19 groundwater species of the genera Niphargus, Niphargellus, Microniphargus and Crangonyx was genetically screened for microsporidian infections. Samples mostly originated from groundwater-dependent spring environments (71%), natural caves (9%) and artificial caverns/tunnels (13%). Amphipod hosts were identified by morphology and/or molecular data, whereas microsporidian parasites were characterised by a genetic screening assay targeting a section of the small subunit rRNA gene. Five microsporidian species (Dictyocoela duebenum; Nosema sp.; Hyperspora aquatica and two undescribed Microsporidium spp.) were revealed from 13 host specimens (Niphargus schellenbergi; N. aquilex lineages B, F and G; Niphargellus arndti). In particular N. schellenbergi was frequently infected with D. duebenum as well as a new and potentially niphargid-specific Nosema sp. identified in Niphargellus arndti. Our results shed further light on the still largely unknown diversity and specificity of microsporidian parasites in groundwater amphipods and subterranean animals in general.

Molecular identification of microsporidian species in patients with epithelial keratitis.

Introduction. Ocular microsporidiosis is a significant emerging infectious disease reported in immunocompromised patients and immunocompetent persons throughout the world.Aim. To identify the pathogens responsible for human keratitis, via corneal scrapings.Methodology. Thirty-three hospitalized patients with epithelial keratitis were examined using staining and DNA sequencing. DNA was extracted from corneal samples and the small-subunit ribosomal RNA gene was amplified by polymerase chain reaction (PCR) and sequenced.Results. Twenty-one samples were positive by staining while PCR generated amplicons in 18 cases. Of the 18 sequences, 16 were identical with, or very similar to, those of Vittaforma corneae (99-100 % similarity) and the remaining two sequences were similar to that of unidentified Microsporidium species deposited in the GenBank.Conclusion. This study has reconfirmed that V. corneae causes epithelial keratitis in humans and that a newly detected Microsporidium species is also involved in microsporidial keratitis as one of the emerging pathogens in Thailand. Ophthalomologists should be aware of microsporidial keratitis in people from Thailand and those from neighbouring countries.

Molecular Characterization and Phylogenetic Analysis of Enterocytozoon bieneusi in Lambs in Oromia Special Zone, Central Ethiopia.

Enterocytozoon bieneusi is the most frequently diagnosed microsporidian species in humans and occurs in a wide range of animals. This study was conducted in Central Ethiopia to determine the prevalence and genotypes of E. bieneusi in lambs in order to evaluate their public health significance. Three hundred eighty nine fecal samples were collected and screened using a nested PCR targeting the internal transcribed spacer (ITS) of the ribosomal RNA gene. All positive PCR products were sequenced to determine the genotypes. E. bieneusi was found in 39 (10.03%) of the lambs. Differences in the infection rates among sex and age groups were not significant (P > 0.05). Five ITS genotypes belonging to three known genotypes BEB6, COS-I, and COS-II, and two novel genotypes (ET-L1 and ET-L2) were identified in lambs. All five genotypes identified in the present study clustered within cattle-specific Group 2 in the ITS phylogenetic tree. This first molecular detection and characterization of E. bieneusi in lambs in Ethiopia has identified the need for further studies in humans and other domestic animals in order to determine the public health significance of E. bieneusi in Ethiopia.

First identification and genotyping of Enterocytozoon bieneusi in humans in Myanmar

BackgroundIntestinal pathogen infections are widespread among impoverished populations. Enterocytozoon bieneusi is the most common pathogen of intestinal microsporidian species in humans worldwide. However, no epidemiological information is available on E. bieneusi infection in humans in Myanmar. The present study comprised the first identification and genotyping of E. bieneusi in humans conducted in Myanmar.ResultsA total of 172 fecal specimens were collected from the Wa people (one each) in four villages of Pangsang Township of the Matman District of Shan State, Myanmar, and each participant completed a questionnaire. E. bieneusi was identified and genotyped using polymerase chain reaction (PCR) and sequence analysis of the internal transcribed spacer (ITS) region of the ribosomal RNA (rRNA) gene. The average prevalence of E. bieneusi was 8.72% (15/172), ranging from 3.85 to 13.89% by village. E. bieneusi infection was not related to any of the risk factors studied. Six genotypes were identified, comprising two known genotypes Peru6 (n = 10) and D (n = 1) and four novel genotypes (MMR23, MMR25, MMR86, and MMR87) (one each), and two people infected with genotype Peru6 were from the same family. A phylogenetic analysis based on a neighbor-joining tree of the ITS sequences of E. bieneusi indicated that all the six genotypes were clustered into group 1.ConclusionsThis is the first identification and genotyping of E. bieneusi in humans in Myanmar. The observations that the two people infected with genotype Peru6 were from the same family, and that all six genotypes obtained in the present study fell into zoonotic group 1, showed the potential for anthropogenic and zoonotic transmissions. The present data argue for the importance of epidemiological control and prevention from medical sectors.

Biodiversity of beet webworm microsporidia in Eurasia

The beet webworm Loxostege sticticalis (LS) has eruptive type of population dynamics and high migratory activity. The LS outbreaks are therefore difficult to predict and the pest belongs to the category of highly dangerous pests. However, during periods of depressions this insect is not observed within the most of its range and is very susceptible to infection by pathogens, including various species of microsporidia, some of which are not specific parasites of the order Lepidoptera. The distribution of LS microsporidia in Eurasia is quite extensive. During the study period of LS from 2003 to 2019, we have found 6 species of microsporidia. The parasite list includes not only species known for Lepidoptera such as Nosema sp. and Vairimorpha thomsoni, but also Vairimorpha (Nosema) ceranae as the typical pathogen from Apis mellifera, as well as Endoreticulatus cf poecilimonae, a pathogen similar to Endoreticulatus poecilimonae from Poecilimon thoracicus. Moreover, two isolates from the genus Tubulinosema identified in LS belong to the group of parasites with a very wide host range, including humans. In laboratory experiments, LS proved high sensitivity to microsporidia N. pyrausta from Ostrinia nubilalis and N. tyriae from Tyria jacobaeae. Its susceptibility to Paranosema locustae from Locusta migratoria has also been discovered.

Impact of Nosema maddoxi on the survival, development, and female fecundity of Halyomorpha halys

Nosema maddoxi Becnel, Solter, Hajek, Huang, Sanscrainte, & Estep, a microsporidian species native to the United States, has been found infecting the invasive brown marmorated stink bug, Halyomorpha halys (Stål). Microsporidian infections in insects often shorten lifespans, decrease fecundity, prolong development, and stunt growth. This study was conducted to determine the impact of N. maddoxi on H. halys fitness. Adult females (2 doses) and nymphs (1 dose) drank suspensions of N. maddoxi spores to promote infection. Adult females receiving a high dose died faster than the controls. Nosema maddoxi infections impacted female egg production and egg viability at both doses compared with the controls. Infections were transmitted to 34.9% of adult males caged with infected females. As the number of days after inoculation increased, infection intensity (# spores found within an infected individual) for both adult treatments transitioned from low-intensity to high-intensity. Infected nymphs died significantly sooner than the controls. Of the treated nymphs, 55.9% died before molting into the fourth instar and only 26.5% eclosed to adults. Nymphal development rate and size were not impacted by N. maddoxi infection. These results indicate that N. maddoxi infection can negatively impact the lifespan of adult females, female fecundity, egg viability, and nymphal survival, which we hypothesize would negatively impact H. halys population densities.

Molecular detection and genotype distribution of Enterocytozoon bieneusi in farmed silver foxes (Vulpes vulpes) and arctic foxes (Vulpes lagopus) in Shandong Province, eastern China.

Enterocytozoon bieneusi is an opportunistic enteric pathogen which can infect a wide range of animal species and humans. It is the most diagnosed species of Microsporidia in humans and has an impact on public health. Many infected animals including foxes may be a potential source for transmitting E. bieneusi to humans. However, limited information is available on the E. bieneusi prevalence and genotypes in farmed foxes in China. Therefore, in the present study, 344 fresh fecal samples were collected from farmed foxes (Vulpes vulpes and Vulpes lagopus) in Shandong Province, and the prevalence and genotypes of E. bieneusi were examined based on sequence analysis of the ribosomal internal transcribed spacer (ITS) region. The overall E. bieneusi prevalence was 9% (31/344); of them, 6.5% (9/138) in farmed silver foxes (V. vulpes) and 10.7% (22/206) in farmed arctic foxes (V. lagopus). Moreover, four known (Hum-q1, NCF2, HND-1, and Type IV) and two novel E. bieneusi genotypes (SDF1 and SDF2) were identified in farmed foxes in the present study. All of the E. bieneusi genotypes belonged to the zoonotic group based on phylogenetic analysis. In addition, 2, 4, 0, and 11 samples were successfully amplified at MS1, MS3, MS4, and MS7 loci, respectively. The present study reveals E. bieneusi prevalence and genotype distribution in farmed foxes in Shandong Province and enlarged the host and geographic information of E. bieneusi in China.

Hardware – wheat pests as elements of the species consortium

Annually in the world, insect pests kill 10–15 % of the crop yield, which in the face of increasing humanity creates a global problem for food security. For the territory of Ukraine in some years, the loss of wheat yields reach 25 % and, in the face of this tendency, real risks to the security of the state are created. Considering that the agrarian trend of the state development has been increasing in recent years, the development of grain-adapted to the ecological and climatic conditions of Ukraine is a priority direction of the development of domestic agriculture. And the development of new environmentally friendly methods of controlling pests of wheat using parasitic organisms will become a fundamentally new and pressing issue in the development of organic production in the country. The general list of pest species that can cause serious wheat yield and forage reduction in Ukraine includes more than 230 names, but only 18 species are able toentail the biggest losses. Among them belong to the order Coleoptera. One insect species is usually a host of several species of mites, nematodes and up to ten species of gregarines and microsporidia. Unfortunately, the taxonomic composition of parasites of phytophagous pets of wheat in Ukraine has not been thoroughly investigated. We studied the interrelation between parasites of 4 phytophagous insects of wheat as well as of 14 insect species found in granaries. The study of the species composition of parasitic organisms of wheat and grain pests is based on the study of the elementary part of the consortium ecosystem, where they act as second order consortia. Phytophagous parasites of wheat and collar pests of cereals, as second-order consorts, have been fragmented worldwide. The purpose of this work is to determine the species composition of gregarine as a component of wheat consortium for their further use in the creation of integrated methods of protection of wheat and products of its processing. As a result, 28 species of gregarines were registered, which belong to seven families (Didymophyidae, Ophryocystidae, Hirmocystidae, Lipotrophidae, Stylocephalidae, Actinocephalidae and Gregarinidae). On the average, 1–2 species of gregarines parasitize in one pest species. 7–10 species of parasites were identified in some insects of the family Tenebrionidae (Tenebrio molitor Linnaeus, 1758 and Tribolium confusum Jacquelin du Val, 1863). Each presented parasite is characteristic of 1–2 species of pests, but there are species of gregarines that infect up to four species of model species, such as Gregarina cuneata Stein, 1848. The largest number of individuals who are infested with gregarines belong to such families as Tenebrionidae and Dermestidae (38,6 % and 36,0 % of the total number of species, respectively). On pests – phytophages parasitizes about 8 species of gregarine families Stylocephalidae, Gregarinidae, Actinocephalidae. For barns pests are recorded barnacles of all 7 families.

Polar tube structure and three polar tube proteins identified from Nosema pernyi

Microsporidian spores contain a single polar filament that is coiled around the interior of the spore. Upon germination the polar tube (post-germination polar filament) is ejected by inversion into a host cell. The sporoplasm flows through the polar tube, directly infecting the cytoplasm of the cell. Various species of microsporidia display differences in the number of coils in the polar filament and in the amino acid sequence of the polar tube proteins (PTPs). Nosema pernyi is a lethal pathogen that causes microsporidiosis in the Chinese oak silkworm, Antheraea pernyi. In this study, we identified three PTPs in N. pernyi using RT-PCR and LC-MS/MS. Polar tube protein 3 was localized in the polar tube using immuno-histochemical staining and an immunofluorescence assay. Co-immunoprecipitation data and LC-MS/MS analysis revealed that some potential proteins, like immune related proteins in A. pernyi may interact with PTP3.