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Isolation and characterization of Bacillus safensis strain NCT49 from a marine fish gut

Authors:

Abstract

As a part of academic lab in bacteriology, marine fish gut sample was collected, serially diluted and plated on to nutrient agar plates. The pure culture was preserved using glycerol stocking in triplicates. This glycerol stocked cultures were revived and used for further morphological and molecular characterization.
National College Culture Collection Centre
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
1
Isolation and characterization of Bacillus safensis strain NCT49 from a marine fish gut
Background
As a part of academic lab in bacteriology, marine fish gut sample was collected, serially diluted
and plated on to nutrient agar plates. The pure culture was preserved using glycerol stocking in
triplicates. This glycerol stocked cultures 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
2
Strain description: NCT49 grows luxuriantly in nutrient agar plate at 37°C in 24 hours. Colonies
are small, white, round with smooth edges. Colonies were on average 0.3mm in size (last quadrant
metrics). Cells are Gram positive, sporulative 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 NCT49 was sensitive to streptomycin (17mm), clindamycin (7mm) and resistant to
neomycin and ampicillin.
16S rRNA gene sequencing:
Based on the colony and biochemical characteristics, the strain NCT49 was identified as Bacillus
safensis. NCT49 was 99.78% similar to Bacillus safensis (GenBank accession number:
NR_041794). 16S rRNA gene sequence of NCT27 can be globally accessed through genbank
accession number OQ271431.
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 1359
positions in the final dataset.
The strain NCT49 was deposited in
National College Culture Collection
Centre (ncccc) and are freely available for
academic and research purposes. The
strain can be obtained here www.ncccc.in
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