• International Journal of Industrial Entomology and Biomaterials
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• v.33 no.2
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• pp.121-131
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• 2016

Ten microsporidian isolates from Samia cynthia ricini, and Antheraea assamensis in India along with a Nosema reference strain (NIK-1s_mys) from B. mori India were characterised morphologically and molecular based tools. The test isolates observed elongated oval in shape while reference strain was oval and ranging from 3.80 to 4.90 m in length and 2.60 to 3.05 m in width. The ribosomal DNA region 'IGS' of test isolates assessed by PCR amplification, followed by cloning and sequencing. IGS sequence and phylogenetic analysis of test microsporidian isolates showed very close relationship with three Nosema references species: N. philosamia, N. antheraea isolated from Philosamia cynthia ricini and Antheraea perny in China respectively and N. disstriae from Malacosma disstriae in Canada. The clustering pattern of dendogram reveals all test isolates appear distinct from Nosema std. (NIK-1s_mys) India used as reference strain in the study. The result suggests IGS indeed a suitable and highly applicable molecular tool for identifying and characterise the microsporidian isolates in similar population.

• International Journal of Industrial Entomology and Biomaterials
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• v.30 no.1
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• pp.6-16
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• 2015

This study established the genetic characterisation of 10 microsporidian isolates infecting non-mulberry silkworms (Antheraea assamensis and Samia cynthia ricini) collected from biogeographical forest locations in the State of Assam, India, using PCR-based markers assays: inter simple sequence repeat (ISSR) and random amplified polymorphic DNA (RAPD). A Nosema type species (NIK-1s_mys) was used as control for comparison. The shape of mature microsporidian spores were observed oval to elongated, measuring 3.80 to $4.90{\mu}m$ in length and 2.60 to $3.05{\mu}m$ in width. Fourteen ISSR primers generated reproducible profiles and yielded 178 fragments, of which 175 were polymorphic (98%), while 16 RAPD primers generated reproducible profiles with 198 amplified fragments displaying 95% of polymorphism. Estimation of genetic distance coefficients based on dice coefficients method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was done to unravel the genetic diversity of microsporidians infecting Indian muga and eri silkworm. The similarity coefficients varied from 0.385 to 0.941 in ISSR and 0.083 to 0.938 in RAPD data. UPGMA analysis generated dendrograms with two microsporidian groups, which appear to be different from each other. Based on Euclidean distance matrix method, 2-dimensional distribution also revealed considerable variability among different identified microsporidians. Clustering of these microsporidian isolates was in accordance with their host and biogeographic origin. Both techniques represent a useful and efficient tool for taxonomical grouping as well as for phylogenetic classification of different microsporidians in general and genotyping of these pathogens in particular.

• International Journal of Industrial Entomology and Biomaterials
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• v.47 no.1
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• pp.1-11
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• 2023

The Indian golden muga silkworm, Antheraea assamensis Helfer is an economically important wild silkworm endemic to Northeastern part of India. In recent years, climate change has posed a threat to muga silk production due to the requirement that larvae be reared outdoors. Since the muga silkworm larvae are exposed to the vagaries of nature, the changing climate has increased the incidence of microbial diseases in the rearing fields. Accurate diagnosis of the disease causing pathogens and its associated epidemiology are prerequisites to manage the diseases in the rearing field. Although conventional microbial culturing methods are widely used to identify pathogenic bacteria, they would not provide meaningful information on a wide variety of silkworm pathogens. The information on use of molecular diagnostic tools in detection of microbial pathogens of wild silk moths is very limited. A wide range of molecular and immunodiagnostic techniques including denaturing gradient gel electrophoresis (DGGE), random amplified polymorphism (RAPD), 16S rRNA/ITSA gene sequencing, multiplex polymerase chain reaction (M-PCR), fluorescence in situ hybridization (FISH), immunofluorescence, and repetitive-element PCR (Rep-PCR), have been used for detecting and characterizing the pathogens of insects with economic significance. Nevertheless, the application of these molecular tools for detecting and typing entomopathogens in surveillance studies of muga silkworm rearing is very limited. Here, we discuss the possible application of these molecular techniques, their advantages and major limitations. These methods show promise in better management of diseases in muga ecosystem.