Detection of Microsporidia Infecting Beet Webworm Loxostege Sticticalis (Pyraloidea: Crambidae) in European Part of Russia in 2006–2008

Malysh, Julia M.; Kononchuk, A.G.; Фролов, А. Н.

doi:10.31993/2308-6459-2019-2(100)-45-51

Cited by 17 publications

(11 citation statements)

References 2 publications

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“…Alive and perished larvae were dissected and inner tissues were processed for molecular genetic analysis. Genomic DNA was extracted as described previously [12]. Degenerate primers BgDNVfor3 (5′-GWMAGGGWGACGAGCAACGTTC-3′) and BgDNVrev3 (5′-RTTTCCCTWGGTTCRTARTTWGGTTC-3′) were designed to span a fragment, ∼800 bp long, corresponding to the non-structural protein within the open reading frame 3 (ORF3) closer to its 3′-end, in the genomes of two densoviruses presented by Genbank accession #AY189948 and #JQ320376.…”

Section: Methodsmentioning

confidence: 99%

Molecular Identification of a Densovirus in Healthy and Diseased Zophobas morio (Coleoptera, Tenebrionidae)

et al. 2019

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Zophobas morio is a tropical darkling beetle which is widely exploited for commercial large-scale insect growing. Outbreaks of a disease may occur causing total devastation of cultures. In the present paper, samples of diseased Z. morio were obtained and used for establishment of a laboratory model as they were found infective to the larvae of the same insect species from another source. It took about 1 month to develop symptoms of acute disease in mid-age larvae and about twice as much when younger larvae were used for infection. Affected larvae perished quickly, and within several days up to 90-100% of the colony could perish. Both in healthy and diseased larvae a virus was detected using PCR with degenerate primers specific for a gene coding for a nonstructural protein (ORF3). The sequenced gene fragment (Genbank accession #MN732869) confirmed allocation of the virus to Densoviridae, with maximal similarity of 97.2% to Blatella germanica densovirus-like virus (#JQ320376) and 66.2% to B. germanica densovirus (#AY189948). Genomic DNA samples of Z. morio larvae from an independent colony devoid of symptoms of a disease were also positive for this virus with a slightly different (99.7% sequence similarity to the former sequence of the Z. morio densovirus) genotype (#MN732870).

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Section: Methodsmentioning

confidence: 99%

Molecular Identification of a Densovirus in Healthy and Diseased Zophobas morio (Coleoptera, Tenebrionidae)

et al. 2019

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“…CHW-2008 Austria Thaumetopoea processionea Insecta EU260046 [ 47 ] Endoreticulatus sp. isolate JMM 2007 Loxostege sticticalis Insecta MK929470 [ 48 ] Endoreticulatus sp. Melnik Euproctis chrysorrhoea Insecta KU900486 [ 43 ] Endoreticulatus sp.…”

Section: Methodsmentioning

confidence: 99%

Metabarcoding reveals low prevalence of microsporidian infections in castor bean tick (Ixodes ricinus)

et al. 2022

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Background Microsporidia is a large group of eukaryotic obligate intracellular spore-forming parasites, of which 17 species can cause microsporidiosis in humans. Most human-infecting microsporidians belong to the genera Enterocytozoon and Encephalitozoon. To date, only five microsporidian species, including Encephalitozoon-like, have been found in hard ticks (Ixodidae) using microscopic methods, but no sequence data are available for them. Furthermore, no widespread screening for microsporidian-infected ticks based on DNA analysis has been carried out to date. Thus, in this study, we applied a recently developed DNA metabarcoding method for efficient microsporidian DNA identification to assess the role of ticks as potential vectors of microsporidian species causing diseases in humans. Methods In total, 1070 (493 juvenile and 577 adult) unfed host-seeking Ixodes ricinus ticks collected at urban parks in the city of Poznan, Poland, and 94 engorged tick females fed on dogs and cats were screened for microsporidian DNA. Microsporidians were detected by PCR amplification and sequencing of the hypervariable V5 region of 18S rRNA gene (18S profiling) using the microsporidian-specific primer set. Tick species were identified morphologically and confirmed by amplification and sequencing of the shortened fragment of cytochrome c oxidase subunit I gene (mini-COI). Results All collected ticks were unambiguously assigned to I. ricinus. Potentially zoonotic Encephalitozoon intestinalis was identified in three fed ticks (3.2%) collected from three different dogs. In eight unfed host-seeking ticks (0.8%), including three males (1.1%), two females (0.7%) and three nymphs (0.7%), the new microsporidian sequence representing a species belonging to the genus Endoreticulatus was identified. Conclusions The lack of zoonotic microsporidians in host-seeking ticks suggests that I. ricinus is not involved in transmission of human-infecting microsporidians. Moreover, a very low occurrence of the other microsporidian species in both fed and host-seeking ticks implies that mechanisms exist to defend ticks against infection with these parasites. Graphical abstract

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“…For DNA analysis, genomic DNA was extracted routinely (Malysh et al., 2019) from spore samples, infected larvae, and experimental egg batches, followed by PCR and amplicon sequencing with 18f:1047r flanking a ca. 850 bp long region of microsporidia small subunit ribosomal RNA (SSU rRNA) gene (Weiss & Vossbrinck, 1999) and NbHKfor1:NpyrHKrev1 primers flanking a 709 bp long region of N. pyrausta hexokinase (Tokarev et al., 2019).…”

Section: Methodsmentioning

confidence: 99%

Transovarial transmission of Nosema pyrausta in three generations of Ostrinia nubilalis in laboratory tests

Grushevaya

Senderskiy

Zubarev

et al. 2021

Entomologia Exp Applicata

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The microsporidium Nosema pyrausta (Paillot) (Nosematidae) is a widespread and persistent entomopathogen with integrated pest management implications. Nosema pyrausta is capable of horizontal and vertical transmissions in Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) populations. After experimental per os infection, N. pyrausta virulence decreased with the instar of the host larvae used. At a dosage of 100,000 spores per fourth instar, 47% pupae and 36% adults survived and gave rise to transovarially infected offspring. The infection persisted through three laboratory generations of the host with prevalence levels ranging between 29 and 92%. Mature microsporidian spores were not found in smears prepared from the eggs laid by infected females, but cryosections demonstrated parasite cells within the host egg’s cytoplasm. A short region of N. pyrausta hexokinase was amplified using cDNA samples from infected females’ eggs, confirming the expression of the parasite’s DNA. Thus, vertical transmission of N. pyrausta is achieved through infection of insect eggs with prespore stages of the parasite.

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Detection of Microsporidia Infecting Beet Webworm Loxostege Sticticalis (Pyraloidea: Crambidae) in European Part of Russia in 2006–2008

Cited by 17 publications

References 2 publications

Molecular Identification of a Densovirus in Healthy and Diseased <b><i>Zophobas morio</i></b> (Coleoptera, Tenebrionidae)

Molecular Identification of a Densovirus in Healthy and Diseased <b><i>Zophobas morio</i></b> (Coleoptera, Tenebrionidae)

Metabarcoding reveals low prevalence of microsporidian infections in castor bean tick (Ixodes ricinus)

Transovarial transmission of Nosema pyrausta in three generations of Ostrinia nubilalis in laboratory tests

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