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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.

The microsporidium Nosema pyrausta is an important mortality factor of the European corn borer, Ostrinia nubilalis. The present study was aimed at N. pyrausta virulence testing to the beet webworm (BW), Loxostege sticticalis. This agricultural pest, L. sticticalis, was highly vulnerable to N. pyrausta. The parasite’s spores were located in salivary glands, adipose tissue, and Malpighian tubules of the infected specimens. Infection was transmitted transovarially through at least 3 laboratory generations, in which BW fitness indices were lower than in the control, and moth emergence and fertility decreased prominently. Transovarial infection was most detrimental to female egg-laying ability, resulting in zero fertility in F3. When propagated in BW, the microsporidium tended to increase its virulence to L. sticticalis, as compared to the Ostrinia isolates. The parasite’s ability to infect this host at low dosages and transmit vertically should guarantee its effective establishment and spread within BW populations. In conclusion, N. pyrausta is a promising agent against BW, which is a notorious polyphagous pest in Eurasia.

Loma acerinae is a xenoma-forming fish microsporidium described from common ruffe Gymnocephalus cernua (Perciformes: Percidae) and also found in Ponto-Caspian gobies (Gobiiformes: Gobiidae). This casts doubt on the strict host specificity of this parasite. The largest subunit RNA polymerase II (rpb1) was used as a genetic marker of the parasite isolated from six host species of Perciformes (G. cernua from the Baltic Sea), Atheriniformes (Atherina boyeri from the Azov Sea) and Gobiiformes (Neogobius spp. and Zosterisessor ophiocephalus from the Black Sea and Ponticola kessleri from the Caspian Sea basin). Two major rpb1 haplogroups were found with 98.5% identity between the groups. Notably, Haplogroup I was associated with Neogobius spp. samples (n = 6) only, whereas Haplogroup II included the samples from other host species (n = 7). These findings confirm the broad distribution and host range of L. acerinae, but also indicate that certain patterns of host-driven intraspecific polymorphism may exist. Furthermore, the study revealed low similarity between the ribosomal RNA gene sequences of L. acerinae and the type species, Loma morhua (as well as other species of the genus). This suggests loose genetic association within the genus, and may raise the need for the taxonomic revision of L. acerinae.

Two species of microsporidia, Nosema apis and Nosema ceranae, occur regularly and cause significant losses in apiculture throughout the world. N. ceranae is thought to be an emerging pathogen of the European honey bee which is replacing N. apis. Microscopic analysis of honey bees collected in Tyumen region, South-Western Siberia, suggested presence of two microsporidial pathogens slightly differing in spore size and shape. PCR detection using species-specific primer sets 312APIS and 218MITOC followed by PCR product sequencing confirmed the diagnosis of N. apis and N. ceranae, respectively. Microsporidia were present in private apiaries through 2008-2010, and among 21 colonies from 7 localities, two colonies were infected with both pathogens, while infections with N. apis only were detected in 8, and with N. ceranae only in 13 colonies. These data suggest that N. ceranae is widely spread in South-Western Siberia alongside with N. apis and is able to persist in the regions with average January temperatures below –18°C.

Chorthippus loratus collected in Krasnodar Territory in 2017 was infected at 15% rate with a microsporidium possessing ovocylindrical binucleate spores, 2.6 × 1.2 μm in size. SSU RNA gene typing (Genbank accession # MH396491) showed its allocation to the genus Liebermannia. Degenerate primers based upon largest subunit RNA polymerase II (RPB1) sequences of closest relatives allowed amplifying the respective gene fragment of Liebermannia sp. (# MH396492). The present finding indicates worldwide distribution of the Liebermannia genus and parasitism in hosts with nonoverlapping geographic ranges (representing Neotropical versus Palearctic fauna), while previous observations were restricted to Acridoidea endemic for South America.

A comparison of the small subunit rRNA gene sequences of the microsporidian isolates from corn borers of the Ostrinia genus from Russia and Belarus indicates that they belong to the Nosema pyrausta species. In order to analyze genetic polymorphism, the nucleotide sequences of the molecular clones of the rRNA intergenic spacer (IGS) of the N. pyrausta isolates from Ostrinia nubilalis and Ostrinia scapulalis (Krasnodar territory), as well as from O. nubilalis (Gomel region, Belarus), are determined. Based on the information obtained and available in Genbank, a high level of divergence of the IGS locus between the isolates of different species and populations of microsporidia, as well as between the molecular clones within individual isolates of parasites, is shown.

A microsporidium was found in a Mediterranean cricket Gryllus bimaculatus from a pet market in the UK and a lab stock at the Moscow Zoo (originating from London Zoo). The spores were ovoid, uninucleate, 6.3 × 3.7 μm in size (unfixed), in packets by of 8, 16, or 32. The spores were easily discharged upon dessication or slight mechanical pressure. The polar tube was isofilar, with 15-16 coils arranged in 1-2 rows. The polaroplast was composed of thin lamellae and occupied about one third of the spore volume. The endospore was 200 nm thick, thinning over the anchoring disc. The exospore was thin, uniform, and with no ornamentation. Phylogenetics based upon small subunit ribosomal RNA (Genbank accession # MG663123) and RNA polymerase II largest subunit (# MG664544) genes placed the parasite at the base of the Trachipleistophora/Vavraia lineage. The RPB1 locus was polymorphic but similar genetic structure and identical clones were found in both isolates, confirming their common geographic origin. Due to in insufficient ultrastructural data and prominent divergence from described species, the parasite is provisionally placed to the collective taxon Microsporidium.

Background. Microsporidia are ubiquitous parasites of animals, most abundant in arthropods and fishes. Many species of these parasites are important from standpoints of medicine, veterinary and agriculture. Microsporidium Nosema pyrausta is an important disease agent in corn borer populations causing adverse effect on host fitness. Genotyping of this parasite is necessary for proper species identification and intraspecific polymorphysm studies. Materials and Methods. Microsporidia-infected larvae of corn borers of the genus Ostrinia were recovered from Krasnodar Territory in Russia and Gomel Region in Belarus. Small subunit ribosomal RNA (SSU rRNA) was amplified and sequenced directly, while intergenic spacer (IGS) was amplified, cloned and sequenced (1-5 clones per sample) for four isolates of microsporidia. Sequences were aligned and compared using standard bioinformatics tools (Clustal W and BLAST). Results. SSU rRNA genotyping showed allocation of all four isolates to N. pyrausta with 100% identity to each other and 99.7% similarity to Nosema bombycis, the type species of the genus Nosema. High levels of IGS sequence variation (61-74%) is observed both between isolates of different species and populations of microsporidia as well as between molecular clones within parasite isolates from individual hosts. Conclusion. N. pyrausta is widespread in corn borer populations and its genetic structure is complicated, as in other species of these parasites. Further studies of molecular markers are needed for genetic differentiation of geographic isolates of N. pyrausta.

The microsporidian genera Nosema and Vairimorpha comprise a clade described from insects. Currently the genus Nosema is defined as having a dimorphic life cycle characterized by diplokaryotic stages and diplosporoblastic sporogony with two functionally and morphologically distinct spore types ("early" or "primary" and "environmental"). The Vairimorpha life cycle, in addition to a Nosema-type diplokaryotic sporogony, includes an octosporoblastic sporogony producing eight uninucleate spores (octospores) within a sporophorous vesicle. Molecular phylogeny, however, has clearly demonstrated that the genera Nosema and Vairimorpha, characterized by the absence or presence of uninucleate octospores, respectively, represent two polyphyletic taxa, and that octosporogony is turned on and off frequently within taxa, depending on environmental factors such as host species and rearing temperature. In addition, recent studies have shown that both branches of the Vairimorpha-Nosema clade contain species that are uninucleate throughout their life cycle. The SSU rRNA gene sequence data reveal two distinct clades, those closely related to Vairimorpha necatrix, the type species for the genus Vairimorpha, and those closely related to Nosema bombycis, the type species for the genus Nosema. Here, we redefine the two genera, giving priority to molecular character states over those observed at the developmental, structural or ultrastructural levels and present a list of revised species designations. Using this approach, a series of species are renamed (combination novum) and members of two genera, Rugispora and Oligosporidium, are reassigned to Vairimorpha because of their phylogenetic position. Moreover, the family Nosematidae is redefined and includes the genera Nosema and Vairimorpha comprising a monophyletic lineage of Microsporidia.

Hexokinase (HK) is a core glycolytic enzyme of Microsporidia which regulates host cell metabolic processes. The goal of the present study was to test for the utility of HK for molecular phylogenetics, species identification and molecular detection of microsporidia in infected insects. HK sequence-based reconstructions were essentially similar to those based upon largest subunit RNA polymerase (RPB1) gene sequences, as well as previously published rRNA gene and genome-based trees. Comparing HK sequences allowed clear differentiation of closely related taxa, such as Nosema bombycis and Nosema pyrausta. In Nosema ceranae, unique SNPs were found for an isolate from wild colonies of the Burzyan dark honey bee as compared with the isolates from domesticated European honey bee. Similarly, in Encephalitozoon cuniculi, HK was as effective as RPB1 for discrimination of isolates belonging to different ITS genotypes. Amplification using species-specific primers flanking short fragments at the 3′-end of HK gene showed the presence of infection in insect tissues infected with N. pyrausta, Nosema ceranae and Paranosema (Antonospora) locustae. For the latter parasite species, HK expression was also demonstrated at early stages of infection using total mRNA extracts of locust larvae. These results indicate the suitability of HK as a novel tool for molecular genetic studies of Microsporidia.

An isolate of the microsporidium Vairimorpha ephestiae (originally isolated from Ephestia kuhniella) from collection of Prof. J. Weiser was propagated in a laboratory culture of Galleria mellonella. Only disporoblastic sporogony was observed and formation of octospores, characteristic of the genus Vairimorpha, never occurred. A partial nucleotide sequence of the small subunit rRNA gene (1247 bp) for this microsporidium showed 100% identity to the homologous sequences of Vairimorpha (Nosema) necatrix (Genbank accession # U11051 and # DQ996241), a microsporidium with a broad host range within the Lepidoptera. Sequence similarity of protein-coding genes (RPB1, HSP70 and actin) between V. ephestiae and V. necatrix was about 98–100%. The level of genetic polymorphism in the RPB1 locus between these two species was essentially the same as between isolates of V. necatrix. It is therefore concluded that V. ephestiae is in fact an isolate of V. necatrix and the former species should be synonymized with the latter. Though described later, V. necatrix has prevailing usage and its precedence over V. ephestiae is proposed to conserve stability and avoid confusion.

Three species of Microsporidia were identified from a population of the beet webworm Loxostege sticticalis at prevalence rates of 35, 4, and 3%. The most prevalent parasite (Tubulinosema sp.) was similar to Tubulinosema acridophagus (99.8% ssrDNA sequence similarity) and was also isolated from the parasitoid Lydella thompsoni (Diptera, Tachinidae) that emerged from the beet webworms. In laboratory assays, spores of this Tubulinosema sp. showed an infection rate of up to 80% for both L. sticticalis and Galleria mellonella larvae. The spores were viable after 12 months of storage in dried infected cadavers. The second most prevalent parasite was closely related to Nosema furnacalis and Nosema granulosis (98.7% similarity). Fresh spores showed a 50% infection rate under laboratory conditions. The third most abundant parasite was identified as the honeybee pathogen Nosema ceranae (100% ssrDNA and 95–100% IGS similarity). In the laboratory, fresh spores of N. ceranae isolated from beet webworm and honey bee were infective to L. sticticalis larvae at the rates of 5 and 2%, respectively.

Populations of European corn borer (Ostrinia nubilalis Hubner) from Krasnodar Territory (Southwestern Russia) become regularly infected with Nosema-like microsporidia. To identify the parasite, it was subjected to electron microscopy and small subunit ribosomal RNA (SSU rRNA) gene sequencing. The spore ultrastructure of the parasite was highly similar to Nosema bombycis from China and Nosema pyrausta from the USA. The nucleotide sequence of SSU rRNA gene was identical to a microsporidium isolated from O. nubilalis in southern France (GenBank accession no. HM566196) and closely related to Nosema bombycis (no. AY209011, 99.7 % sequence similarity) from Bombyx mori of Chinese origin and N. pyrausta (no. AY958071) from O. nubilalis of North American origin. As the molecular haplotype of SSU rRNA is fixed for the parasite infecting O. nubilalis across Europe and N. pyrausta was initially described in France as Perezia pyraustae (Paillot CR Acad Sci Paris 185: 673–675, 1927), we conclude that the parasite examined under the present study correspond to the type isolate of N. pyrausta. The microsporidium from O. nubilalis in North America (no. AY958071) corresponds therefore to a closely related, yet distinct haplotype.

We examined whether maize offers enemy-free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid-related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis' immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels.