Application of Fluorescence in situ hybridization-Flow cytometry (FISH-FCM) technique to detect and quantify Vibrio cholerae population from different geographic regions

Lidita Khandeparker (CSIR-National Institute of Oceanography)
Dattesh V. Desai (CSIR-National Institute of Oceanography, Goa, India)
Arga Chandrashekar Anil (CSIR-National Institute of Oceanography)
S. S. Sawant (CSIR-National Institute of Oceanography, Goa, India)
K. Venkat (CSIR-National Institute of Oceanography, Goa, India)
Kaushal Mapari (CSIR-National Institute of Oceanography, Goa, India)
Zuliza Jolkifli (Fisheries Ecology and Oceanography Section, Department of Fisheries, Ministry of Industry and Primary Resources, Brunei Darussalam)
Noorizan Abd. Karim (Fisheries Ecology and Oceanography Section, Department of Fisheries, Ministry of Industry and Primary Resources, Brunei Darussalam)
Hikmah Thoha (Research Center for Oceanography, Indonesian Institute of Sciences, Jakarta, Indonesia)
Hadiyanto Hadiyanto (Research Center for Oceanography, Indonesian Institute of Sciences, Jakarta, Indonesia)
Soukaseum Dalasane (Port and Navigation Division, Department of Waterways, Ministry of Public Works and Transport, Vientiane, Lao PDR)
Kongngeun Chounlamountry (Lao National Mekong Committee Secretariat, Ministry of Natural Resources and Environment, Khounboulom Road, Vientiane, Lao PDR)
Myint Myint Khaing (Remote Sensing Department, Mandalay Technological University, Mandalay, Myanmar)
Jenelle Clarisse Dungca (The Marine Science Institute, College of Science, University of the Philippines, Diliman, Quezon City, The Philippines)
Rhodora Azanza (The Marine Science Institute, College of Science, University of the Philippines, Diliman, Quezon City, The Philippines)
Chin Sing Lim (St John’s Island National Marine Laboratory, Tropical Marine Science Institute, National University of Singapore, Singapore)
Koh Siang Tan (St John’s Island National Marine Laboratory, Tropical Marine Science Institute, National University of Singapore, Singapore)
Sumana Kajonwattanakul (Department of Marine and Coastal Resources, Phuket Marine Biological Center (Bangkok Office), Bangkok, Thailand)
Ratchanee Phuttapreecha (Department of Marine and Coastal Resources, Marine and Coastal Resources Research Center (Southern Part), Songkhla, Thailand)
Hoang Mai Le (Northern Center for Integrated Coastal Management and Planning, Vietnam Administration of Seas and Islands, Ha Noi, Vietnam)

ASEAN Journal on Science and Technology for Development
Vol. 35 No. 1-2 (2018): Extent of transfer of alien invasive organisms in South/Southeast Asia regio

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.