My Ssec Capstone Project Intended Category

Intended Category

Intended Category: NEW SPECIES
Title: ‘Stenotrophomonas johoriensis’ sp. nov., a new bacteral species isolated from marine ballast water
Author’s name: Salleh.N.A., Hamidun Bunawan and Farah Nadiah Rosli
Institutional Affiliation: National University of Malaysia
Corresponding author: Hamidun Bunawan
Institute of Systems Biology (INBIOSIS)
Universiti Kebangsaan Malaysia
43600 UKM Bangi
Selangor Darul Ehsan
MALAYSIA
Tel: +603 8921 4554
Fax: +603 8921 3398
[email protected]
Abstract
We report the main characteristic of ‘Stenotrophomonas johoriensis’ sp. nov, a new species within the genus Stenotrophomonas that was isolated from marine ballast water from Port Tanjung Pelepas, Johor, Malaysia.
Keywords: ‘Stenotrophomonas johoriensis’ ,ballast water, Port Tanjung Pelepas
The deballasting of ballast water from ships that contain a diverse community of organisms acts as a vehicle for the global distribution of pathogens, invasive species and waterborne diseases which potentially impose an environmental and health threat to coastal regions 1. As a part to study the diversity of bacteria community in ballast water, we isolated a new bacterium , strain ptp 12.25 from ballast water tank in Port Tanjung Pelepas, Johor, Malaysia in May 2017. By using systematic matrix-assisted desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) screening on a MicroFlex spectrometer (Bruker Daltonics, Bremen, Germany) no identification of bacteria was obtained.
Seawater sample was serially diluted in sterile saline solution and dilutions were spread on marine agar medium. The culture was incubated at 30oC for 2 days in an aerobic condition. Morphological characteristics of single colony were as follows; white in colour and circular. Meanwhile gram staining results indicate gram negative bacterium with rod-shaped and non-spore forming. Catalase and oxidase were positive.

Molecular identification for complete 16S rRNA gene was sequence using 27f-1525r 2 primer and a 3730-XL Data Analyzer (1st Base, Singapore Science Park II, Singapore) sequencer. The obtained sequence was 83% similar to the 16S rRNA gene sequence of Stenotrophomonas maltophilia strain N97b3 (GenBank accession no. KM894189.1) 3, which was pylogenetically closest species with standing in nomenclature (Fig. 1)
According to the 16S rRNA gene sequence similarity in NCBI database, we propose that strain ptp 12.25 is a representative of a new species within genus Stenotrophomonas. As such, we propose the name ‘Stenotrophomonas johoriensis’ sp. nov. as acknowledgement to the place origin of isolated bacteria.
MALDI-TOF MS spectrum accession number
KIV
30861001684020
FIG. 1. Pylogenetic tree highligthing position of ‘Stenotrophomonas johoriensis’ (underlined) relative to other pylogenetically closet members of Stenotrophomonas genus. Number at node is percentage of bootsrap value obtained by repeating analysis 1000 times to generate majority consesus tress. Only value ;70% were displayed. Scale bar represents 2% nucleotide sequence divergence.
Nucleotide sequence accession number
The 16S rRNA gene sequence of the strain Stenotrophomonas johoriensis was deposited in NCBI GenBAnk under accession number MH635159.

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Deposit in culture collection
Strain Stenotrophomonas johoriensis was deposited in Collection de Souches de I’Unite des Rickettsies (CSUR, WDCM 875) (KIV)
Acknowledgements
This research was funded by the Malaysian Ministry of Science Technology and Innovation (MOSTI) as part of grant (04-01-02-SF1244).

Conflict of Interest
None declared
Reference
1 Ng. C., Le. T., Goh. S.G., Liang. L., Kim. Y., Rose. J.B., Hoong. K. G.Y. A Comparison of Microbial Water Quality and Diversity for Ballast and Tropical Harbor Waters.PloS ONE 2015;10(11): e143123.
2 Frank. J.A., Reich. C.I., Sharma. S., Weisbaum. J.S., Wilson. B.A., Olsen. G.J. Critical evaluation of two primers commonly used for amplification of bacterial 16S rRNA genes. Appl Environ Microb. 2008;74:2461–247
3Gish. W., States. D.J. “Identification of protein coding regions by database similarity search.” Nature Genet. 1993; 3:266-272.

4 Hasegawa M., Kishino H., and Yano T. (1985). Dating the human-ape split by a molecular clock of mitochondrial DNA. Journal of Molecular Evolution 22:160-174
5 Felsenstein J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39:783-791

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