• International Journal of Industrial Entomology and Biomaterials
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• v.16 no.1
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• pp.29-35
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• 2008

The midgut epithelium of Indian honey bee Apis cerana indica is consist of digestive cells and small regenerative cells. The regenerative cells are placed in the nests scattered among the digestive cells. During metamorphosis the midgut of Apis cerana indica is remodeled. The larval midgut epithelium and muscular sheath digested partially at the end of larval period and thrown out in the form of debris in the lumen. The new epithelium is formed by the proliferation of the regenerative cells and during pupation reorganization of midgut layer occurs. The ultrastuctural studies shows that the regenerative cells are in contact with degenerative cells by the cytoplasmic extension which have many septed and gap junctions in the fifth instar larvae. In developing pupae reorganization of the midgut epithelium is continued whereas in the pharate adult the midgut wall shows, characteristic of adult midgut epithelium with pycnotic nuclei in some cells.

• BMB Reports
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• v.36 no.5
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• pp.508-513
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• 2003

The bacterium Bacillus thuringiensis produces toxin inclusions that are deleterious to target insect larvae. These toxins are believed to interact with a specific receptor protein(s) that is present on the gut epithelial cells of the larvae. In various insect species (in particular those belonging to the lepidopteran class), aminopeptidase N (APN) is one of the two receptor proteins that are considered to be involved in toxin-receptor interactions. However, in mosquitoes, the nature and identity of the receptor protein is unknown. Here, using RT-PCR, we identified two isoforms of the APN transcripts in the Aedes aegypti mosquito larval midgut. These results are congruent with a previous report of multiple isoforms of the APN gene expression in lepidopteran larvae. Which of the two isoforms (or other yet unidentified receptor proteins) is involved in the killing of mosquito larvae remains to be elucidated.

• BMB Reports
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• v.40 no.5
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• pp.783-790
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• 2007

Receptor binding plays an important role in determining host specificity of the Bacillus thuringiensis Cry $\delta$-endotoxins. Mutations in domains II and III have suggested the participation of certain residues in receptor recognition and insect specificity. In the present study, we expressed the cloned domain II-III fragment of Cry4Ba and examined its binding characteristics to mosquito-larval midgut proteins. The 43-kDa Cry4Ba-domain II-III protein over-expressed in Escherichia coli as inclusion bodies was only soluble when carbonate buffer, pH 10.0 was supplemented with 4M urea. After renaturation via stepwise dialysis and subsequent purification, the refolded domain II-III protein, which specifically reacts with anti Cry4Ba-domain III monoclonal antibody, predominantly exists as a $\beta$-sheet structure determined by circular dichroism spectroscopy. In vitro binding analysis to both histological midgut tissue sections and brush border membrane proteins prepared from susceptible Aedes aegypti mosquito-larvae revealed that the isolated Cry4Ba-domain II-III protein showed binding functionality comparable to the 65-kDa full-length active toxin. Altogether, the data present the 43-kDa Cry4Ba fragment comprising domains II and III that was produced in isolation was able to retain its receptor-binding characteristics to the target larval midgut proteins.

• International Journal of Industrial Entomology and Biomaterials
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• v.22 no.2
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• pp.59-64
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• 2011

Protein kinase C (PKC) is involved in many cellular signaling pathways, it participates in many physiological processes, such as cell cycle, growth, proliferation, differentiation and apoptosis. To investigate the effect of PKC on the silkworm midgut tissue infection of Bombyx mori parvo-like virus (BmPLV), a B. mori atypical protein kinase C (BmaPKC) gene was cloned from larval midgut tissue, expressed in E. coli and purified. Additionally, the BmPLV susceptible silkworm strain and resistant silkworm strain were used to test the effect of the B. mori infection on BmPLV. The result showed that BmaPKC encodes a predicted 586 amino acid protein, which contains a C-terminal kinase domain and an N-terminal regulatory domain. The maximum expression amount of the soluble (His)6-tagged fusion protein was detected after 0.8 mmol/L IPTG was added and cultured at $21^{\circ}C$. The (His) 6-tagged fusion protein revealed about 73 kDa molecular weight which confirmed by western blot and mass spectrography. Furthermore BmaPKC protein were detected at 0-72 h post-infection in BmPLVinfected larval midgut tissue, western blot showed that as time went on, the expression of BmaPKC increased gradually in susceptible strain, the expression quantity on 72 h is 5 times of 0 h. However, in resistant strain, the expression quantity is slightly lower than susceptible strain. But no significant change in resistant strain was observed as time went on. The available data suggest that BmaPKC may involve in the regulation of BmPLV proliferation.

• The Korean Journal of Zoology
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• v.25 no.2
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• pp.63-70
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• 1982

The activity, properties, and distribution of midgut protease during metamorphosis in Pieris rapae L. are determined using spectrophotometer, ultracentrifuge and agar gel electrophoresis. Proteolytic activity of midgut reaches the peak just before ecdysis in 5th instar and prepupal stages each but 1 day after ecdysis in pupal stage. Also, optimum pH of midgut protease is pH 8.0 in 5th instar stage, pH 6.5 in prepupal stage, and pH 8.5 immediately before emergence respectively. Protease is found mostly in midght tissue in 5th instar stage but thereafter until just before emergence the enzyme only in lumen contents, suggesting that protease is synthesized in midgut tissue during larval stage and then released into lumen during pupation period.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.423-433
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• 2019

Midgut microbiota is known to play a fundamental role in the biology and physiology of the agricultural pest, Spodoptera litura. This study reports the difference in the larval midgut microbiota of field-caught and laboratory-reared populations of S. litura by performing 16S rDNA amplicon pyrosequencing. Field populations for the study were collected from castor crops, whereas laboratory-reared larvae were fed on a regular chickpea based diet. In total, 23 bacterial phylotypes were observed from both laboratory-reared and field-caught caterpillars. Fisher's exact test with Storey's FDR multiple test correction demonstrated that bacterial genus, Clostridium was significantly abundant (p < 0.05) in field-caught larvae of S. litura as compared to that in the laboratory-reared larvae. Similarly, bacterial genera, such as Bradyrhizobium, Burkholderia, and Fibrisoma were identified (p < 0.05) predominantly in the laboratory-reared population. The Bray-Curtis dissimilarity matrix depicted a value of 0.986, which exhibited the maximum deviation between the midgut microbiota of the laboratory-reared and field-caught populations. No significant yeast diversity was seen in the laboratory-reared caterpillars. However, two yeast strains, namely Candida rugosa and Cyberlindnera fabianii were identified by PCR amplification and molecular cloning of the internal transcribed space region in the field-caught caterpillars. These results emphasize the differential colonization of gut residents based on environmental factors and diet.

• International Journal of Industrial Entomology and Biomaterials
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• v.23 no.1
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• pp.107-113
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• 2011

Infection effect of $Nosema$ $bombyics$ on the midgut of silkworm $Bombyx$ $mori$ and subsequent appearance of spores and the performance of larvae was studied. Autopsy of larvae showed white pustules on the surface of midgut at 5 days of post infection (pi). At later stage, important organs like midgut, silk gland and gonads reduced in size and all these organs showed white pustules. Light microscope observation of pustules revealed enormous spores. Spore multiplication was at a faster rate in young larvae. Infection of the adult larvae resulted in pebrinized pupa and moths. Larval weight, cocoon weight and cocoon shell ratio reduced as the post infection period increased. Transverse sections of midgut showed $N.$ $bombycis$ infection limited to a few columnar cells at 3-5 days of pi. At 7 days pi, cell volume increased, cells were swollen and elongated. Heavily infected cells looked like sacks filled with parasite and the apical region of certain cells were bulging into the gut lumen. Later at 8-9 days of pi, spores or its developing stages leaked into the lumen either freely or enclosed within the globules of host cytoplasm. Besides columnar cells, development of $N.$ $bombycis$ was observed in the regenerative cells and rarely in goblet cells. Development of $N.$ $bombycis$ was also observed in both longitudinal and circular muscles at the late pi period. The histopathological changes, deformities and spore production time in the host were all influenced by the spore dosage and age of the host.

• International Journal of Industrial Entomology and Biomaterials
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• v.4 no.2
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• pp.127-131
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• 2002

The qualitative and quantitative changes of bacterial flora associated with the Pure Mysore (Multivoltine) and NB$_4$D$_2$ (Bivoltine) silkworm (Bombyx modi L.) midgut during third, fourth and fifth instars were studied. Larvae reared on mulberry leaves were dissected and their midgut bacterial populations were enumerated through serial dilution technique and after 72 hrs of incubation period at 28 $\pm$ 1$^{\circ}C$, the bacterial population was estimated. The results showed a highest mean value of 15$\times$10/ sup 6/ sup 6/ CFU/g and 28$\times$10/ sup 6/ CFU/g in Pure Mysore and NB$_4$D$_2$races, respectively, in midgut tissue of fifth instar larvae. The natural epiphytic microflora of mulberry leaves fed during the respective instars was also studied and found maximum 14$\times$10$^3$ CFU/g in leaves fed in third instars, followed by 5.3$\times$10$^3$ CFU/g and 2.1$\times$10$^3$ CFU/g in leaves fed during fourth and fifth instars, respectively. The bacterial flora colonized in midgut was found to be elaborating amylase, caseinase, gelatinase, lipase and urease enzymes. The highest percentages of isolates were amylase producers followed by protein and lipid splitters in Pure Mysore, whereas in NB$_4$D$_2$ protein splitter were dominated followed by lipase and amylase producers in NB$_4$D$_2$. The results indicate that the natural microflora may play a vital role in the digestion of ingested food materials in silkworms.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.13-18
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• 2003

Allatotropin is a 13-residue amidated neuropeptide isolated from pharate adult heads of the tobacco hornworm, Manduca serta and strongly stimulates biosynthesis of juvenile hormones in adults, but not larval, lepidopteran corpora allata. From a Bombyx mori midgut cDNA library, a cDNA that encodes a 130-amino-acid polypeptide containing M. sexta allatotropin sequence was isolated. The B. mori allatotropin cDNA consists of 1196 nucleotides. (omitted)

• Proceedings of the Korean Society of Applied Entomolohy Conference
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• 2002.05a
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• pp.177-177
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• 2002