Application of Fluorescence in situ hybridization-Flow cytometry (FISH-FCM) technique to detect and quantify Vibrio cholerae population from different geographic regions
AJSTD Vol 35(1-2) cover
PDF

Keywords

Fluorescence in situ hybridization (FISH)
Flow cytometry (FCM)
bacteria
vibrio cholerae
bioinvasion

Abstract

Rapid and species-specific detection, and quantification of pathogenic bacteria are fundamental for monitoring and assessment of the risk they pose to any ecosystem. The study employed Vibrio cholerae, a human pathogen responsible for the life-threatening diarrhoeal disease, cholera and one among the most unwanted from marine bioinvasion point of view. The present study coupled fluorescence in situ hybridization (FISH) technique, a powerful tool in molecular phylogenetic discrimination, with flow cytometry (FCM), a technique used for rapid and accurate quantification of both viable but non-cultivable and non-viable microorganisms. The FISH-FCM technique was used for the first time to quantify V. cholerae (includes cultivable and non-cultivable) from different geographic regions of Southeast Asia (Brunei, Indonesia, Lao PDR, Myanmar, Philippines, Singapore, Thailand, Vietnam) and India (Goa, west coast of India). The data acquired from the analyses provides a snap shot view of the total bacterial abundance with special reference to V. cholerae. As the method developed, it was evaluated with bacterial samples collected from different sites in Southeast Asia and India, and the application of this technique to different geographical regions appears feasible. Considering that the continuous growth of the shipping industry and ballast water as one of the primary vectors responsible for the global transport of pathogenic microorganisms, the risk they present needs immediate attention. This technique will be useful in the quick and accurate detection of specific pathogens. It may also provide significant insights to quarantine measures for Ballast Water Management.

https://doi.org/10.29037/ajstd.495
PDF

References

Anil AC, Venkat K, Sawant SS, Dileepkumar M, Dhargalkar VK, Ramaiah N, Harkantra SN, Ansari ZA. 2002. Marine bioinvasion: Concern for ecology and shipping. Current Science 83(3):214–219.

Azanza RV, Austero NM, Dungca JCR, Caspe FJO, Khandeparker L. 2018. Phytoplankton and bacterial communities in South Harbour, Manila Bay, Philippines. ASEAN Journal on Science and Technology for Development 35(1–2):107–113.

Carlton JT, Geller JB. 1993. Ecological roulette: the global transport of non-indigenous marine organisms. Science 261:78–82.

Czechowska K, Johnson DR, van der Meer JR. 2008. Use of flow cytometric methods for single-cell analysis in environmental microbiology. Current Opinion in Microbiology 11:205–212.

Davey HM, Jones A, Shaw AD, Kell DB. 1999. Variable selection and multivariate methods for the identification of microorganisms by flow cytometry. Cytometry 35(2):162–168.

Drake LA, Doblin MA, Dobbs FC. 2007. Potential microbial bioinvasions via ships’ ballast water, sediment, and biofilm. Marine Pollution Bulletin 55(7):333–341.

D’Costa PM, Anil AC, Patil JS, Hegde S, D’Silva MS, Chourasia M. 2008. Dinoflagellates in a mesotrophic, tropical environment influenced by monsoon. Estuarine, Coastal and Shelf Science 77:77–90.

Emami K, Askari V, Ullrich M, Mohinudeen K, Anil AC, Khandeparker L, Burgess JG, Mesbahi E. 2012. Characterization of bacteria in ballast water using MALDI-TOF mass spectrometry. PloS One 7(6):38515.

Fernandes SO, Kirchman DL, Michotey VD, Bonin PC, LokaBharathi PA. 2014. Bacterial diversity in relatively pristine and anthropogenicallyinfluenced mangrove ecosystems (Goa, India). Brazilian Journal of Microbiology 45:1161–1171.

Finkelstein RA. 1996. Cholera, Vibrio cholerae O1 and O139, and other pathogenic Vibrios. In: Baron S, editor. Medical Microbiology. 4th ed. Galveston (TX): University of Texas Medical Branch at Galveston. Chapter 24.

Gil AI, Louis VR, Rivera ING, Lipp E, Huq A, Lanata CF, Taylor DN, Cohen ER, Choopun N, Sack RB, Colwell RR. 2004. Occurrence and distribution of Vibrio cholerae in the coastal environment of Peru. Environmental Microbiology 6:699–706.

GloBallast. 2002. The GEF/UNDP/IMO Global Ballast Water Management Programme (GloBallast). Available from http://www.globallast.imo.org >

Hallegraeff GM, Bolch CJ. 1992. Transport of diatom and dinoflagellate resting spores in ships' ballast water: implications for plankton biogeography and aquaculture. Journal of Plankton Research 14(8):1067–1084.

Hewitt C, Campbell M. 2010. The relative contribution of vectors to the introduction and translocation of marine invasive species. Australian Department of Agriculture Fisheries, and Forestry, Canberra, Australia.

Huq A, Small EB, West PA, Huq MI, Rahman R, Colwell RR. 1983. Ecological relationships between Vibrio cholerae and planktonic crustacean copepods. Applied and Environmental Microbiology 45:275–283.

Huq A, Xu B, Chowdhury MAR, Islam MS, Montilla R, Colwell RR. 1996. A simple filtration method to remove plankton associated Vibrio cholerae in raw water supplies in developing countries. Applied and Environmental Microbiology 62:2508–2512.

Huq A, Haley BJ, Taviani E, Chen A, Hasan NA, Colwell RR. 2012. Detection, isolation, and identification of Vibrio cholerae from the environment. Current Protocols in Microbiology; CHAPTER: Unit6A.5. doi:10.1002/9780471729259.mc06a05s26.

Ivanov V. 2000. Monitoring the bacterial neuston. In: Stopa PJ, Bartoszcze MA, editors. Rapid methods for analysis of biological materials in the environment. Netherlands: Springer. p. 67–72.

Joachimsthal EL, Ivanov V, Tay SL, Tay JH. 2004. Bacteriological examination of ballast water in Singapore Harbour by flow cytometry with FISH. Marine Pollution Bulletin 49(4):334–343.

Khandeparker R, Verma P, Deobagkar D. 2011. A novel halotolerant xylanase from marine isolate Bacillus subtilis cho40: gene cloning and sequencing. New Biotechnology 28:814–821.

Khandeparker L, Anil AC. 2013. Association of bacteria with marine invertebrates: implications for ballast water management. EcoHealth 10(3):268–276.

Khandeparker L, D'Costa PM, Anil AC, Sawant SS. 2014. Interactions of bacteria with diatoms: Influence on natural marine biofilms. Marine Ecology 35:233–248.

Khandeparker L, Anil AC, Naik SD, Gaonkar CC. 2015. Daily variations in pathogenic bacterial populations in a monsoon influenced tropical environment. Marine Pollution Bulletin 96:337–343.

Khandeparker L, Eswaran R, Gardade L, Kuchi N, Mapari K, Naik SD, Anil AC. 2017a. Elucidation of the tidal influence on bacterial populations in a monsoon influenced estuary through simultaneous observations. Environmental Monitoring and Assessment 189(41):1–17.

Khandeparker L, Kuchi N, Kale D, Anil AC. 2017b. Microbial community structure of surface sediments from a tropical estuarine environment using next generation sequencing. Ecological Indicators 74:172–181.

Kim BH, Lee MK. 2012. A case of bacteremia due to Microbacterium oleivorans identified by 16S rRNA sequencing analysis. Korean Journal of Clinical Microbiology 15(3):110–113.

Li X, Qin L. 2005. Metagenomics-based drug discovery and marine microbial diversity. Trends in Biotechnology 23(11):539–543.

López-Amorós R, Castel S. Comas-Riu J, Vives-Rego J. 1997. Assessment of E. coli and Salmonella viability and starvation by confocal laser microscopy and flow cytometry using rhodamine 123, DiBAC4 (3), propidium iodide, and CTC. Cytometry 29(4):298–305.

Lyon WJ. 2001. TaqMan PCR for Detection of Vibrio cholera O1, O139, Non-O1, and Non-O139 in pure cultures, raw oysters, and synthetic seawater. Applied and Environmental Microbiology 67(10):4685–4693.

Maskey RP, Li FC, Qin S, Heinz HF, Laatsch H. 2003. Chandrananimycins A-C: Production of novel anticancer antibiotics from a marine Actinomadura sp. Isolate M048 by variation of medium composition and growth conditions. Journal of Antibiotics 56:622–629.

McCallum HI, Kuris A, Harvell CD, Lafferty KD, Smith GW, Porter J. 2004. Does terrestrial epidemiology apply to marine systems? Trends in Ecology and Evolution 19(11):585–591.

Mimura H, Katakura R, Ishida H. 2005. Changes of microbial populations in a ship’s ballast water and sediments on a voyage from Japan to Qatar. Marine Pollution Bulletin 50(7):751–757.

Moriarty DJW. 1998. Control of luminous Vibrio species in penaeid aquaculture ponds. Aquaculture 164(1-4):351–358.

Mourino-Pérez RR, Worden AZ, Azam F. 2003. Growth of Vibrio cholerae O1 in red tide waters off California. Applied and Environmental Microbiology 69:6923–6931.

Not F, Simon N, Biegala IC, Vaulot D. 2002. Application of fluorescent in situ hybridization coupled with tyramide signal amplification (FISH-TSA) to assess eukaryotic picoplankton composition. Aquatic Microbial Ecology 28(2):157–166.

Oren A. 2004. Prokaryote diversity and taxonomy: current status and future challenges. Philosophical Transactions of the Royal Society of London 359:623–638.

Philippine Ports Authority. 2015. Ports services handbook: South Harbour. Manila: Philippine Ports Authority.

Rehnstam-Holm A –S, Godhe A, Härnström K, Raghunath P, Saravanan V, Collin B, Karunasagar I, Karunasagar I. 2010. Association between phytoplankton and Vibrio spp. along the southwest coast of India: a mesocosm experiment. Aquatic Microbial Ecology 58:127–139.

Roszak DB, Grimes DJ, Colwell RR. 1984. Viable but nonrecoverable stage of Salmonella enteritidis in aquatic systems. Canadian Journal of Microbiology 30(3):334–338.

Ruiz GM, Rawlings TK, Dobbs FC, Drake LA, Mullady T, Huq A, Colwell RR. 2000. Global spread of microorganisms
by ships. Nature 408(6808):49–50.

Sack RB, Siddique AK, Longini IM, Nizam A, Yunus M, Islam MS, Morris JG, Ali A, Huq A, Nair B, Qadri F, Faruque SH, Sack DA, Colwell RR. 2003. A 4-year study of the epidemiology of Vibrio cholerae in four rural areas of Bangladesh. Journal of Infectious Diseases 187(1):96–101.

Schoemar S, Yanagi T. 2001. Seasonal variation of water characteristics in the northern coastal area of Java. La mer 39:77–85.

Shvalov AN, Soini JT, Surovtsev IV, Kochneva GV, Sivolobova GF, Petrov AK, Maltsev VP. 2000. Ndividual Escherichia coli cells studied from light scattering with the scanning flow cytometer. Cytometry 41(1):41–45.

Sims D, Brettin T, Detter JC, Han C, Lapidus A, Copeland A, Del Rio TG, Nolan M, Chen F, Lucas S, Hope T, Cheng J-F, Bruce D, Goodwin L, Pitluck S, Ovchinnikova G, Pati A, Ivanova N, Mavromatis K, Chen A, Palaniappan K, D’haeseleer P, Chain P, Bristow J, Eisen JA, Markowitz V, Hugenholtz P, Schneider S, Göker M, Pukall R, Kyrpides NC, Klenk H-P. 2009. Complete genome sequence of Kytococcus sedentarius type strain (541T). Standards in Genomic Science 1:12–20.

Siregar TH, Priyanto N, Putri AK, Rachmawati N, Triwibowo R, Dsikowitzky L, Schwarzbauer J. 2016. Spatial distribution and seasonal variation of the trace hazardous element contamination in Jakarta Bay, Indonesia. Marine Pollution Bulletin 110(2):634–646.

Smith LD, Wonham MJ, McCann LD, Reid DM, Carlton JT, Ruiz GM. 1996. Biological invasions by nonindigenous species in United States waters: quantifying the role of ballast water and sediments. Parts I and II. The National Biological Invasions Shipping Study II (NABISS II) US Coast Guard and the US Department of Transportation, Washington DC (Rep. No. CG-D-02-97, Gov. Access. No. ADA321543).

Sugita H, Hirose Y, Matsuo N, Deguchi Y. 1998. Production of the antibacterial substance by Bacillus sp. Strain NM 12, an intestinal bacterium of Japanese coastal fish. Aquaculture 165(3-4):269–280.

Sun B, Mouland R, Way C, Rivkin RB. 2010. Redistribution of heterotrophic prokaryotes through ballast water: a case study from the west coast of Canada. Aquatic Invasions 5(1):5–11.

Tang KW. 2005. Copepods as microbial hotspots in the ocean: effects of host feeding activities on attached bacteria. Aquatic Microbial Ecology 38:31–40.

Troussellier M, Courties C, Zettelmaier S. 1995. Flow cytometric analysis of coastal lagoon bacterioplankton and picophytoplankton: fixation and storage effects. Estuarine, Coastal and Shelf Science 40(6):621-633.

Tsukamura M. 1971. Proposal of a new genus, Gordonia, for slightly acid-fast organisms occurring in sputa of patients with pulmonary disease and in soil. Journal of General Microbiology 68:15–26.

Williams RJ, Griffiths FB, Van der Wal EJ, Kelly J. 1988. Cargo vessel ballast water as a vector for the transport of non-indigenous marine species. Estuarine, Coastal and Shelf Science 26(4):409–420.
Creative Commons License

This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.

Downloads

Download data is not yet available.