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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, 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 bact...

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Main Authors: Rahman, Mohammad Mustafizur, Izuddin, Mohd Haikal, Khan, Najmus Sakib, Mohamad, Mohd Azrul Naim, John, Akbar
Format: Book Chapter
Language:English
Published: Springer 2017
Subjects:
SH Aquaculture. Fisheries. Angling
Online Access:http://irep.iium.edu.my/61358/1/1.pdf
http://irep.iium.edu.my/61358/
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spelling my.iium.irep.613582018-01-18T01:21:10Z http://irep.iium.edu.my/61358/ 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 Mohamad, Mohd Azrul Naim John, Akbar SH Aquaculture. Fisheries. Angling DNA barcoding along with molecular phylogeny can be used to taxonomic identification, characterization, 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 (i) identify the observed bacterial isolates by comparing the partial sequence from an unknown sample to a collection of sequences from known reference samples, and (ii) know the taxonomic and phylogenetic identity of identified bacteria in the fish, and (iii) know the abundance of fish bacteria in the Kuantan River. For this study, three commercially important fish namely Pristipomoides filamentosus, Cyclocheilichtys apogon and Labiobarbus festivus were captured with gill nets from Kuantan River, Malaysia. Bacteria from skin, gill and gut in fish were culture at 35°C for 24 h in both nutrient and marine agar. Bacterial DNA were extracted using Bacterial Genomic DNA Isolation Kit following manufacturer’s specifications. Isolated DNA were 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. Neighbour-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 Cyclocheilichtys 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 (5.12×103) and P. filamentosus (5.20×103) (Fig 3). Overall, the highest bacterial abundance was observed in fish gut (6.62×103), followed by fish gill (5.78×103) and fish skin (4.60×103). Springer 2017-11 Book Chapter REM application/pdf en http://irep.iium.edu.my/61358/1/1.pdf Rahman, Mohammad Mustafizur and Izuddin, Mohd Haikal and Khan, Najmus Sakib and Mohamad, Mohd Azrul Naim and John, Akbar (2017) DNA barcoding and molecular phylogeny of indigenous bacteria in fishes from a tropical tidal river in Malaysia. In: DNA barcoding and molecular phylogeny. Springer, pp. 1-31. (In Press)
institution Universiti Islam Antarabangsa Malaysia
building IIUM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider International Islamic University Malaysia
content_source IIUM Repository (IREP)
url_provider http://irep.iium.edu.my/
language English
topic SH Aquaculture. Fisheries. Angling
spellingShingle SH Aquaculture. Fisheries. Angling
Rahman, Mohammad Mustafizur
Izuddin, Mohd Haikal
Khan, Najmus Sakib
Mohamad, Mohd Azrul Naim
John, Akbar
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, 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 (i) identify the observed bacterial isolates by comparing the partial sequence from an unknown sample to a collection of sequences from known reference samples, and (ii) know the taxonomic and phylogenetic identity of identified bacteria in the fish, and (iii) know the abundance of fish bacteria in the Kuantan River. For this study, three commercially important fish namely Pristipomoides filamentosus, Cyclocheilichtys apogon and Labiobarbus festivus were captured with gill nets from Kuantan River, Malaysia. Bacteria from skin, gill and gut in fish were culture at 35°C for 24 h in both nutrient and marine agar. Bacterial DNA were extracted using Bacterial Genomic DNA Isolation Kit following manufacturer’s specifications. Isolated DNA were 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. Neighbour-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 Cyclocheilichtys 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 (5.12×103) and P. filamentosus (5.20×103) (Fig 3). Overall, the highest bacterial abundance was observed in fish gut (6.62×103), followed by fish gill (5.78×103) and fish skin (4.60×103).
format Book Chapter
author Rahman, Mohammad Mustafizur
Izuddin, Mohd Haikal
Khan, Najmus Sakib
Mohamad, Mohd Azrul Naim
John, Akbar
author_facet Rahman, Mohammad Mustafizur
Izuddin, Mohd Haikal
Khan, Najmus Sakib
Mohamad, Mohd Azrul Naim
John, Akbar
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 2017
url http://irep.iium.edu.my/61358/1/1.pdf
http://irep.iium.edu.my/61358/
_version_ 1643615985974902784
score 13.211869

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