• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.1
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• pp.63-69
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• 2005

While observing silkworm larval samples received from field, microsporidian spores formed within the haemocytes of silkworm haemolymph were observed. The spores of microsporidian sp. were purified and characterized for morphological characters viz., size, shape as well as serological affinity with different Nosema spp. (M$_{11}$ and M$_{12}$). The infectivity of the isolated spores to silkworm was also studied. The microsporidian sp. was found to be highly pathogenic to silkworm, B. mori. The isolated microsporidian sp. was designated as NIK-5hm, which formed ovocylindrical spore in the haemocytes of silkworm and differed in spore size (length, 4.55 $\mu$m & width, 2.10 $\mu$m) and shape from Nosema bombycis (NIK-ls), NIK-2r (Nosema sp. Mysore [3.6 & 2.8 $\mu$m]), NIK-3h (Nosema sp. M$_{11}$ [3.8 & 1.8 $\mu$m]), NIK-4m (Nosema sp. M$_{12}$ [5.0 & 2.1 $\mu$m]) and Lb$_{ms}$ (Nosema sp. in Lamerine breed of silkworm [4.36 & 2.14]). In immonological test (Latex agglutination test), the isolated microsporidian spores did not react with antibody sensitized latex particles of N. bombycis, M$_{11}$, M$_{12}$ and Lb$_{ms}$ and thus are different type of microsporidian sp., parasitic to silkworm, Bombyx mori L.

• International Journal of Industrial Entomology and Biomaterials
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• v.9 no.2
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• pp.265-267
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• 2004

The silkworm, Bombyx mori L. is prone to infection of various pathogenic organisms. Pebrine, one of the deadliest disease of silkworm caused by highly virulent parasitic microsporidian, Nosema bombycis has been understood since long. Infections of the disease range from chronic to highly virulent and can result in complete lose to the sericulture industry. Several strains and species of microsporidians have since been isolated from the infected silkworms. A new microsporidian spore was isolated from Lamerin breed of the silkworm B. mori have been studied under scanning electron microscope, found to be different in spore size (length 4.36$\pm$0.06 ${\mu}{\textrm}{m}$, width 2.14$\pm$0.01${\mu}{\textrm}{m}$) and shape (ova cylindrical with slight depression) from standard strain N. bombycis (length 3.08$\pm$0.21 ${\mu}{\textrm}{m}$, width 2.01$\pm$0.05 ${\mu}{\textrm}{m}$ and ovidal respectively). In immunological test, the silkworm breed Lamerin isolated micrisporidian spore does not react to different antibody (N. bombycis, M$_{11}$ and M$_{12}$) sensitized latex particle and thus appeared to be a different strain of microsporidian parasitic to the Lamerin breed of the silkworm B. mori.i.i.

• International Journal of Industrial Entomology and Biomaterials
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• v.10 no.2
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• pp.143-145
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• 2005

Lamerin breed of the silkworm, Bombyx mori L. for Northeastern India hosts a vertically transmitted microsporidian parasite for generations, which does not harm significantly the cocoon production. The transversally infected progenies do not exhibit marked external sign or symptom. The microspordian causes inapparent infection and over $(80\%)$ of the infected progeny survives and spin cocoons. There is possibility of co-existence between the breed and the associated micosporidian parasite. To evaluate the impact of the microsporidian on breed the present study was conducted in respect of tranovarial transmitted (observed as T1), secondarily infected (observed as T2) and healthy silkworm (observed as T3). The larval and pupal mortality was $12\%$ and $6\%$ in T1 and $10\%$ and $3\%$ in T2 batch, while in case of T3 batch there was no mortality. Significant changes were also observed in single cocoon weight, single shell weight, denier, reelibility, raw silk recovery $\%$ and neetness. There is no significant impact of the infection on the fecundity and hatchability. The hatchability of the eggs laid by healthy or infected moths are equall as much as control but the progeny had the infection transmitted from the parent.

• International Journal of Industrial Entomology and Biomaterials
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• v.12 no.1
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• pp.41-43
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• 2006

Lamerin breed of the silkworm Bombyx mori L. is the only mulberry silkworm breed which survives with an associated microsporidian $(Lb_{ms})$ infection from generations. This present comparative investigation has produced information on the influence of $Lb_{ms}$ on the growth and development of the breed. The $Lb_{ms}$ transmits tranovarially as well as perorally at low level. Larvae developed from the eggs laid by female, infected with $Lb_{ms}$ were as normal as larvae developed from the eggs laid by healthy female. In infected tissues early developmental stages and mature spores located intracellularly but did not show hypertrophy of cells of the host. The meronts and sporonts measures $0.46{\pm}0.25,\;1.11{\pm}0.05{\mu}m$ in length and $0.32{\pm}0.05,\;0.78{\pm}0.10{\mu}m$ in width respectively. The mature spore measures $4.36{\pm}0.06{\mu}m$ in length and $2.14{\pm}0.01{\mu}m$ in width.

• 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.29 no.2
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• pp.169-178
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• 2014

In this study, we investigated genetic diversity of 22 microsporidian isolates infecting tropical tasar silkworm, Antheraea mylitta collected from various geographical forest locations in the state of Jharkhand, India, using polymerase chain reaction (PCR)-based marker assay: random amplified polymorphic DNA (RAPD). A type species, NIK-1s_mys was used as control for comparison. The shape of mature microsporidians was found to be oval to elongate, measuring 3.80 to $5.10{\mu}m$ in length and 2.56 to $3.30{\mu}m$ in width. Of the 20 RAPD primers screened, 16 primers generated reproducible profiles with 298 polymorphic fragments displaying high degree of polymorphism (97%). A total of 14 RAPD primers produced 45 unique putative genetic markers, which were used to differentiate the microsporidians. Calculation of genetic distance coefficients based on dice coefficient method and clustering with un-weighted pair group method using arithmetic average (UPGMA) analysis was conducted to unravel the genetic diversity of microsporidians infecting tasar silkworm. The similarity coefficients varied from 0.059 to 0.980. UPGMA analysis generated a dendrogram with four microsporidian groups, which appear to be different from each other as well as from NIK-1s_mys. Two-dimensional distribution based on Euclidean distance matrix also revealed considerable variability among different microsporidians identified from the tasar silkworms. Clustering of few microsporidian isolates was in accordance with the geographic origin. The results indicate that the RAPD profiles and specific/unique genetic markers can be used for differentiating as well as to identify different microsporidians with considerable accuracy.

• 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.11 no.2
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• pp.81-86
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• 2005

The mode of transmission, effect on fecundity, hatching and tissues specificity of a microsporidian $(Lb_{ms})$ recovered from Lamerin breed of the silkworm Bombyx mori L. was studied and compared with standard strain Nosema bombycis. Peroral inoculation of $Lb_{ms}$ or N. bombycis to zeroday of $4^{th}$ instar larvae of silkworm was the most suitable method for producing information on development of stage specific mortality, pupation and obtaining infected adults for transovarial transmission studies. It was observed that pupal mortality, the percentage of moths emerged and the percentage of moths infected were significantly high in N. bombycis infected batches as compared $(Lb_{ms})$ in all the three tested breeds of the silkworm. However no significant difference was observed in larval mortality. The fecundity and hatchability was not affected significantly in $(Lb_{ms})$ infected adults, however significant reduction in egg production, fecundity, hatchability and increased egg retention was observed in mother moths infected with N. bombycis. The $(Lb_{ms})$ is transmitted both horizontally and vertically at lower rate due to its low rate of proliferation. The trans ovarial transmission of $(Lb_{ms})$ to the $F_1$ progeny generation through eggs averaged only $61.33\pm5.10\%$ whereas N. bombycis was transmitted at $100\%$. The $(Lb_{ms})$ had low oral infectivity and low transovarial transmission in silkworm B. mori.

• International Journal of Industrial Entomology and Biomaterials
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• v.11 no.2
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• pp.107-111
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• 2005

Impact of microsporidian infection and season on reproductive potential of Bombyx mori L. was investigated in the laboratory. Microsporidian infection significantly (P<0.01) reduced fecundity and hatching and increased sterility and mortality of eggs. Among the microsporidia, Nosema sp. 2 infected silkworm produced eggs with least fecundity and hatching percent as well as highest dead and sterile eggs followed by Nosema sp. 1 and N. bombycis. Microsporidia, in general, significantly reduced fecundity and hatching percent of eggs and increased number of dead and sterile eggs in all the three seasons except N. bombycis in July - August (S3) and Nosema sp. 2 in January February (S1). Since, seed production is the anchor sheet of mulberry sericulture, coefficient of egg lying is considered as an important aspect and the industry quite often facing shortage of disease free layings. The present study indicates that B. mori is more susceptible to microsporidia during S2 followed by S3 and S1 and Nosema sp2. is most virulent followed by Nosema spl. and N. bombycis.

• Journal of Sericultural and Entomological Science
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• v.33 no.1
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• pp.27-31
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• 1991

Microsporidian spores were isolated from yellow-hind winged arctiid, Eilema griseola (Hubner), and cutworm, Agrotis tokionis Butler, and their morphological characteristics and their indectivity to silkworms were investigated. The shape of the microsporidian isolates was oval and the sige of the spore isolated from yellow-hind winged was measured in 2.6$\times$1, 5$\mu$ and that of the isolated from cutworm was measured in 3.7$\times$20.$\mu$. Both the isolated microsporidians showed the infectivity to silkworms, but the isolates did transovarian transmission in silkworms and the silkworm moth infected with the isolate from yellow-hind winged laid eggs irregularly.