DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia

DNA barcoding along with molecular phylogeny can be used to taxonomic identification, characterization, and discovery of species and understand molecular relationships especially in term of species divergence. Thus, they facilitate biodiversity studies. Some studies have addressed DNA barcoding of b...

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Main Authors:	Rahman, Mohammad Mustafizur, Izuddin, Mohd Haikal, Khan, Najmus Sakib, John, Akbar, Naim, Mohd Azrul
Format:	Book Chapter
Language:	English
Published:	Springer 2018
Subjects:	SH Aquaculture. Fisheries. Angling
Online Access:	http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/1/69476_DNA%20barcoding%20and%20molecular%20phylogeny.pdf

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recordtype	eprints
spelling	iium-694762019-01-28T08:12:13Z http://irep.iium.edu.my/69476/ DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia Rahman, Mohammad Mustafizur Izuddin, Mohd Haikal Khan, Najmus Sakib John, Akbar Naim, Mohd Azrul SH Aquaculture. Fisheries. Angling DNA barcoding along with molecular phylogeny can be used to taxonomic identification, characterization, and discovery of species and understand molecular relationships especially in term of species divergence. Thus, they facilitate biodiversity studies. Some studies have addressed DNA barcoding of bacterial samples from various sources. Unfortunately, the DNA barcoding of fish bacterial diversity has yet been studied especially in the tropical tidal river. Therefore, a study was conducted to (1) identify the observed bacterial isolates by comparing the partial sequence from an unknown sample to a collection of sequences from known reference samples, (2) know the taxonomic and phylogenetic identity of identified bacteria in the fish, and (3) know the abundance of fish bacteria in the Kuantan River. For this study, three commercially important fish, namely, Pristipomoides filamentosus, Cyclocheilichthys apogon, and Labiobarbus festivus were captured with gill nets from Kuantan River, Malaysia. Bacteria from skin, gill, and gut in fish were cultured at 35 �C for 24 h in both nutrient and marine agars. Bacterial DNA were extracted using Bacterial Genomic DNA Isolation Kit following manufacturer’s specifications. IsolatedDNAwere quantified in NanoDrop 2000v and gel eluded in 1.5% agarose gel and visualized under gel visualizer. PCR products were out source sequenced at First Base Sdn.bhd using ABI sequencer under Sanger sequencing method. Sequences were trimmed using Sequence Scanner 2.0V. The aligned sequences were inspected by eye and edited to remove ambiguities based on PHRED scores and chromatogram. Fully aligned sequences were subjected to BLAST for nucleotide similarity search against 16S rRNA database. Best matched species was selected based on BLAST results and lowest genetic distance between known and unknown nucleotides. Genetic distance (sequence divergences) were calculated using the K2P (Kimura two parameter) distance model. Neighbor-joining (NJ) trees of K2P distances were created to provide a graphic representation of the patterning of divergence between species. This method identified a total of 11 fish bacteria, which taxonomically classified into Enterobacteriales, Pseudomonadales, Actinomycetales, and Bacillales. The range of pair-wise genetic distances between species of Enterobacteriales was lower than Bacillales. Similarly, within group mean genetic distance of Enterobacteriales (0.010) was lower than Bacillales (0.055). These results indicate that the identified bacteria species under Enterobacteriales are more closely related than bacteria species under Bacillales. The mean genetic distance between groups were genetically almost equally closed which was confirmed by overall mean diversity. Out of 11 species 7 were identified in Cyclocheilichthys apogon, 8 in Labiobarbus festivus, and 7 in Pristipomoides filamentosus. Overall mean bacterial abundance (CFU/g) were higher in C. apogon (6.68 � 103) compare to L. festivus (5.12103) and P. filamentosus (5.20103). Overall, highest bacterial abundances were observed in fish gut (6.62103), followed by fish gill (5.78103) and fish skin (4.60*103). Springer 2018 Book Chapter PeerReviewed application/pdf en http://irep.iium.edu.my/69476/1/69476_DNA%20barcoding%20and%20molecular%20phylogeny.pdf Rahman, Mohammad Mustafizur and Izuddin, Mohd Haikal and Khan, Najmus Sakib and John, Akbar and Naim, Mohd Azrul (2018) DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia. In: DNA barcoding and molecular phylogeny. Springer, Switzerland, pp. 351-367. ISBN 978-3-319-90679-9 https://link.springer.com/chapter/10.1007/978-3-319-90680-5_22 https://doi.org/10.1007/978-3-319-90680-5
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	SH Aquaculture. Fisheries. Angling
spellingShingle	SH Aquaculture. Fisheries. Angling Rahman, Mohammad Mustafizur Izuddin, Mohd Haikal Khan, Najmus Sakib John, Akbar Naim, Mohd Azrul DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
description	DNA barcoding along with molecular phylogeny can be used to taxonomic identification, characterization, and discovery of species and understand molecular relationships especially in term of species divergence. Thus, they facilitate biodiversity studies. Some studies have addressed DNA barcoding of bacterial samples from various sources. Unfortunately, the DNA barcoding of fish bacterial diversity has yet been studied especially in the tropical tidal river. Therefore, a study was conducted to (1) identify the observed bacterial isolates by comparing the partial sequence from an unknown sample to a collection of sequences from known reference samples, (2) know the taxonomic and phylogenetic identity of identified bacteria in the fish, and (3) know the abundance of fish bacteria in the Kuantan River. For this study, three commercially important fish, namely, Pristipomoides filamentosus, Cyclocheilichthys apogon, and Labiobarbus festivus were captured with gill nets from Kuantan River, Malaysia. Bacteria from skin, gill, and gut in fish were cultured at 35 �C for 24 h in both nutrient and marine agars. Bacterial DNA were extracted using Bacterial Genomic DNA Isolation Kit following manufacturer’s specifications. IsolatedDNAwere quantified in NanoDrop 2000v and gel eluded in 1.5% agarose gel and visualized under gel visualizer. PCR products were out source sequenced at First Base Sdn.bhd using ABI sequencer under Sanger sequencing method. Sequences were trimmed using Sequence Scanner 2.0V. The aligned sequences were inspected by eye and edited to remove ambiguities based on PHRED scores and chromatogram. Fully aligned sequences were subjected to BLAST for nucleotide similarity search against 16S rRNA database. Best matched species was selected based on BLAST results and lowest genetic distance between known and unknown nucleotides. Genetic distance (sequence divergences) were calculated using the K2P (Kimura two parameter) distance model. Neighbor-joining (NJ) trees of K2P distances were created to provide a graphic representation of the patterning of divergence between species. This method identified a total of 11 fish bacteria, which taxonomically classified into Enterobacteriales, Pseudomonadales, Actinomycetales, and Bacillales. The range of pair-wise genetic distances between species of Enterobacteriales was lower than Bacillales. Similarly, within group mean genetic distance of Enterobacteriales (0.010) was lower than Bacillales (0.055). These results indicate that the identified bacteria species under Enterobacteriales are more closely related than bacteria species under Bacillales. The mean genetic distance between groups were genetically almost equally closed which was confirmed by overall mean diversity. Out of 11 species 7 were identified in Cyclocheilichthys apogon, 8 in Labiobarbus festivus, and 7 in Pristipomoides filamentosus. Overall mean bacterial abundance (CFU/g) were higher in C. apogon (6.68 � 103) compare to L. festivus (5.12103) and P. filamentosus (5.20103). Overall, highest bacterial abundances were observed in fish gut (6.62103), followed by fish gill (5.78103) and fish skin (4.60*103).
format	Book Chapter
author	Rahman, Mohammad Mustafizur Izuddin, Mohd Haikal Khan, Najmus Sakib John, Akbar Naim, Mohd Azrul
author_facet	Rahman, Mohammad Mustafizur Izuddin, Mohd Haikal Khan, Najmus Sakib John, Akbar Naim, Mohd Azrul
author_sort	Rahman, Mohammad Mustafizur
title	DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
title_short	DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
title_full	DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
title_fullStr	DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
title_full_unstemmed	DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia
title_sort	dna barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in malaysia
publisher	Springer
publishDate	2018
url	http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/ http://irep.iium.edu.my/69476/1/69476_DNA%20barcoding%20and%20molecular%20phylogeny.pdf
first_indexed	2023-09-18T21:38:37Z
last_indexed	2023-09-18T21:38:37Z
_version_	1777412989677207552

DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia

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