Isolation and characterization of Bordetella hinzii strain NCT19 from a human microbiome

Abstract

As a part of academic lab in microbiology, human skin microbiome samples were swabbed onto a nutrient agar media. Morphologically distinct colonies were purified using quadrant streaking. The pure culture was preserved using glycerol stocking in triplicates. One of these glycerol stocks was revived and used for further morphological and molecular characterization.

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National College Culture Collection Centre www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
1
N4C
Isolation and characterization of Bordetella hinzii strain NCT19 from a human
microbiome
Background
As a part of academic lab in microbiology, human skin microbiome samples were swabbed onto
a nutrient agar media. Morphologically distinct colonies were purified using quadrant streaking.
The pure culture was preserved using glycerol stocking in triplicates. One of these glycerol stocks
was 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.

National College Culture Collection Centre www.ncccc.in
PG & Research Department of Biotechnology and Microbiology
National College, Tiruchirappalli- 620 001, Tamil Nadu, India
2
N4C
Strain description: NCT19 grows stringently in nutrient agar plate at 37°C in 24 hours. Colonies
are small, round, milky white, shiny with smooth edges and are 0.4mm in diameter (last quadrant
metrics). Cells are Gram negative, non-motile rods. Biochemical reaction implies Indole positive,
methyl red positive, voges proskauer positive and citrate positive. The strain is positive for
amylase production and negative for lipases. Triple Sugar Iron implies acid butt and slant with no
gas production. Catalase negative and oxidase negative. The strain NCT19 was sensitive to
rifampicin, penicillin and streptomycin and erythromycin.
16S rRNA gene sequencing:
Based on the colony, biochemical and molecular characteristics, the strain NCT19 was identified
as Bordetella hinzii. The 16S rRNA gene sequences of NCT19 was 99.35% similar Bordetella
hinzii (GenBank accession number:
NR_027537) and can be globally
accessed through genbank accession
number OQ255726.
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 1376
positions in the final dataset.
The strain NCT19 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