Whole-cell fluorescence in situ hybridization of harmful diatom pseudo-nitzschia (bacilliariophyceae) in Kuching Estuary

Yong, Hwa Lin (2012) Whole-cell fluorescence in situ hybridization of harmful diatom pseudo-nitzschia (bacilliariophyceae) in Kuching Estuary. [Project Report] (Unpublished)

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Abstract

Pennate diatom of the genus Pseudo-nitzschia is recognized as causative organisms producing domoic acid (DA), a neurotoxin associated with amnesic shellfish poisoning (ASP). Conventional methods using light and electron microscopes such as TEM and SEM are undoubtedly useful for Pseudo-nitzschia species identification. Nevertheless, this requires taxonomic expertise, and EM preparations are laborious. Wholecell fluorescence in-situ hybridization (FISH) has been developed using the molecular approach as a rapid detection tool that eases the detection of Pseudo-nitzschia in natural population. In this study, whole-cell FISH was applied to investigate the distribution of Pseudo-nitzschia species in Kuching estuary. Field sampling was undertaken at Santubong and Samariang estuaries fortnightly from September 2011 till April 2012. The results showed that extremely low abundance of Pseudo-nitzschia cells in both locations. Largesubunit ribosomal RNA oligonucleotide probes for P. circumpora (L-S-Pcir-52-A-18), P. caciantha (L-SPcac-38-A-18), and P. pungens (L-S-Ppun-405-A-18) were designed in silico. Synthesized P. pungens and P. caciantha probes were subsequently optimized using centrifugation-based whole-cell FISH. The probes were tested on culture and spike natural samples, and the results showed that both probes are species-specific. This molecular species detection technique is proven to be useful particularly in harmful algal monitoring.

Item Type: Project Report
Additional Information: Project Report (B.Sc.) -- Universiti Malaysia Sarawak, 2012.
Uncontrolled Keywords: Porphyra--Physiology, Bangiaceae, Pseudo-nitzschia, domoic acid (DA), amnesic shellfish poisoning (ASP), whole-cell FISH, LSU rRNA ligonucleotide probe, FSTS, 2012, Universiti Malaysia Sarawak, UNIMAS, universiti, university, Borneo, Malaysia, Sarawak, Kuching, Samarahan, IPTA, education, undergraduate, research
Subjects: Q Science > Q Science (General)
Q Science > QH Natural history > QH301 Biology
Q Science > QR Microbiology
Divisions: Academic Faculties, Institutes and Centres > Faculty of Resource Science and Technology
Depositing User: Karen Kornalius
Date Deposited: 21 Jan 2015 08:31
Last Modified: 18 May 2015 02:52
URI: http://ir.unimas.my/id/eprint/6293

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