Whole-cell fluorescence in situ hybridization of harmful diatom pseudo-nitzschia (bacilliariophyceae) in Kuching Estuary
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 sp...
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| Format: | Final Year Project Report |
| Language: | English |
| Published: |
Universiti Malaysia Sarawak, UNIMAS
2012
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| Online Access: | http://ir.unimas.my/id/eprint/6293/8/Whole-cell%20Fluorescence%20in%20situ%20Hybridization%20of%20Harmful%20Diatom%20Pseudo-nitzschia%20%28Bacillariophyceae%29%20in%20Kuching%20Estuary.pdf http://ir.unimas.my/id/eprint/6293/ |
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| Summary: | 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. |
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