Sample Collection
Healthy and infected fifth instar larvae of A. proylei were collected from Regional Sericulture Research Station, Mantripukhri, Manipur during summer season of 2019. The details of the location are furnished in Table 1. Healthy tasar silkworms and disease silkworms showing the symptoms of Tiger band disease were collected from the above mentioned area (Fig. 1 a & b). The infected larvae suffering from Tiger band disease were recognized from their black band pattern similar to tiger stripes appearing on the larva (Shantibala, Fraser, et al., 2018). Proper sterilized equipment such as forceps, scissors and hand gloves were used while collecting the insect sample. The collected samples were stored in an ice cold storage box. Samples were brought from the field and surface sterilization was done with 70% ethanol by submersion and rinsing three times using sterile distilled water prior to dissection. The gut tissue of larvae and midgut was dissected out in a sterile environment using dissection scissors. The collected intestinal contents will be immediately stored at 50% glycerol at -80⁰C for future use (Broderick, Raffa, Goodman, & Handelsman, 2004). Three larvae from each location were taken for gut dissection. Individual larvae were used for the isolation of gut bacteria. After isolation of gut bacteria the larvae were transferred to laboratory conditions maintained at 26 ±1º C and 70 % RH at Animal Resources Division Laboratory, Institute of Bioresources and Sustainable Development, Takyelpat, Manipur, India.