Evo 40 ep

Morphological characteristics of isolates were assessed by scanning electron microscopy (SEM). Bacterial colonies were inoculated in ISP-2 medium and incubated at 28 ± 2 °C for 7 days. Cells were centrifuged (Eppendorf, USA) at 8000× g for 10 min and pellet was resuspended in 2%–5% gluteraldehyde (Sigma, Burlington, VT, USA) prepared in 0.1M phosphate buffer, pH 7.2. After incubating samples for 30 min, supernatant was discarded and pellet was resuspended in 1% osmium tetraoxide (Sigma, Burlington, VT, USA), incubated for 1 h and centrifuged at 5000 × g. To the pellet, sterile water was added and centrifuged twice for 10 min at 5000 × g. For dehydration, the pellet was resuspended in 35% ethanol for 10 min, 50% ethanol for 10 min, 75% ethanol for 10 min, 95% ethanol for 10 min, and a final wash with 100% ethanol for 10 min. For SEM analysis, sterilized aluminum stubs and cover slips were inserted into the SCA plates at an angle of about 45 °C. The plates with stubs and coverslips were incubated at 37 °C for 24 h to check any contamination. After 24 h, isolates were introduced along the line where the surface of the stub met the agar medium and incubated at 28 ± 2 °C for 7 days. The stubs were then carefully removed and coated under vacuum, with a film of gold for 25–30 min and viewed on the scanning electron microscope (Zeiss Evo 40 EP, Germany).

Surface and cross-section morphologies of the prepared membranes were observed by scanning electron microscopy (SEM) coupled with EDX analyzer with SEM EVO40EP from Zeiss, Germany. The cross-sections were obtained by fracturing the samples in liquid nitrogen. All samples were first covered with a thin layer of gold before study.

Strain MN 2(6) was isolated from the solitary wasp mud nest collected from Dehradun, India (Kumar et al., 2012a (link)). Cultural characteristics of the strain MN 2(6) was examined every day grown on various International Streptomyces project (ISP) media (Shirling and Gottlieb, 1966 (link)). Micromorphology and sporulation were observed under light microscope by the inclined coverslip technique (Williams et al., 1989 ) on ISP-4 medium after incubating at 27°C for 7 days. The spore chain morphology and spore surface ornamentation were examined by scanning electron microscopy (SEM) (Zeiss EVO 40 EP) of 15-day old cultures grown on ISP-4 according to the method described previously (Kumar et al., 2011 (link)). Physiological characteristics were examined according to the methods described in the ISP (Shirling and Gottlieb, 1966 (link)) and Bergey's Manual of Systematic Bacteriology (Locci, 1989 ). Resistance to some antibiotics was detected by disc diffusion method. The isomeric forms of diaminopimelic acid (DAP) and the diagnostic sugar in the whole-cell hydrolysates were determined as described in MTCC Laboratory manual, IMTECH (1998 ), Chandigarh, India.

The surface morphology of membranes was analyzed by SEM analysis using Carl Zeiss EVO^® 40 EP (Germany). Prior to measurements, the samples were subjected to carbon coating deposition.