Isolation and characterization of Bacillus stercoris strain NCT6 from air samples of lab-II Background

Abstract

As a part of academic lab in bacteriology, air sampled culturable bacterial community were sampled using settle plate technique. Following incubation, morphologically unique colonies were segregated via quadrant streaking. These pure cultures were preserved using glycerol stocking in triplicates. One of these glycerol stocks were revived and used for further morphological and molecular characterization.

Full text

National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
N4C
Isolation and characterization of Bacillus stercoris strain NCT6 from air samples of lab-II
Background
As a part of academic lab in bacteriology, air sampled culturable bacterial community were
sampled using settle plate technique. Following incubation, morphologically unique colonies
were segregated via quadrant streaking. These pure cultures were preserved using glycerol
stocking in triplicates. One of these glycerol stocks were revived and used for further
morphological and molecular characterization.
Work plan
Bergey’s manual was used to determine the lowest possible taxonomic hierarchy and the strain
was commercially outsourced for bidirectional 16S rRNA gene sequencing for species level
resolution.
Results

National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
N4C
Strain description: NCT6 grows well in nutrient agar plate at 37°C in 24 hours. Colonies are
large, white, round with smooth edges and swarms
heavily in nutrient agar. Colonies are 0.8mm in size
(last quadrant metrics). Cells are Gram positive,
spore forming, and non-motile rods. Biochemical
reaction implies Indole negative, methyl red positive,
voges proskauer negative and citrate positive. Triple
Sugar Iron implies acid butt and alkaline slant with
no gas production. Catalase positive and oxidase
positive. The strain NCT6 was sensitive
streptomycin.
16S rRNA gene sequencing: Based on the colony
and biochemical characteristics, the strain NCT6 was
identified as Bacillus stercoris. NCT6 was 99.5%
similar to Bacillus stercoris (GenBank accession
number: NR_181952). 16S rRNA gene sequence of
NCT6 can be globally accessed through genbank
accession number OQ271448.
The Neighbor-Joining method tree with percentage of
replicate trees in which the associated taxa clustered
together in the bootstrap test (500 replicates) are
shown. The tree is drawn to scale, with branch
lengths in the same units as those of the evolutionary
distances used to infer the phylogenetic tree. The
evolutionary distances were computed using the
Maximum Composite Likelihood method and are in
the units of the number of base substitutions per site.
This analysis involved 11 nucleotide sequences. All
ambiguous positions were removed for each
sequence pair (pairwise deletion option). There were
a total of 1399 positions in the final dataset.
The strain NCT6 was deposited in National College Culture Collection Centre (ncccc) and are
freely available for academic and research purposes. The strain can be order here www.ncccc.in.