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.