Antibiotic resistance and molecular typing among cockle (Anadara granosa) strains of Vibrio parahaemolyticus by polymerase chain reaction (PCR)-based analysis

Sahilah Abd. Mutalib, R. A S Laila, H. Mohd Sallehuddin, Osman Hassan, Aminah Abdullah, A. Ahmad Azuhairi

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Genomic DNA of Vibrio parahaemolyticus were characterized by antibiotic resistance, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis. These isolates originated from 3 distantly locations of Selangor, Negeri Sembilan and Melaka (East coastal areas), Malaysia. A total of 44 (n = 44) of tentatively V. parahaemolyticus were also examined for the presence of toxR, tdh and trh gene. Of 44 isolates, 37 were positive towards toxR gene; while, none were positive to tdh and trh gene. Antibiotic resistance analysis showed the V. parahaemolyticus isolates were highly resistant to bacitracin (92 %, 34/37) and penicillin (89 %, 33/37) followed by resistance towards ampicillin (68 %, 25/37), cefuroxime (38 %, 14/37), amikacin (6 %, 2/37) and ceftazidime (14 %, 5/37). None of the V. parahaemolyticus isolates were resistant towards chloramphenicol, ciprofloxacin, ceftriaxone, enrofloxacin, norfloxacin, streptomycin and vancomycin. Antibiogram patterns exhibited, 9 patterns and phenotypically less heterogenous when compared to PCR-based techniques using ERIC- and RAPD-PCR. The results of the ERIC- and RAPD-PCR were analyzed using GelCompare software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the V. parahaemolyticus isolates into 6 clusters and 21 single isolates at a similarity level of 80 %. While, RAPD-PCR with primer Gen8 discriminated the V. parahaemolyticus isolates into 11 clusters and 10 single isolates and Gen9 into 8 clusters and 16 single isolates at the same similarity level examined. Results in the presence study demonstrated combination of phenotypically and genotypically methods show a wide heterogeneity among cockle isolates of V. parahaemolyticus.

Original languageEnglish
Pages (from-to)649-659
Number of pages11
JournalWorld Journal of Microbiology and Biotechnology
Volume30
Issue number2
DOIs
Publication statusPublished - Feb 2014

Fingerprint

Arcidae
Cardiidae
Vibrio parahaemolyticus
Molecular Typing
Microbial Drug Resistance
Polymerase Chain Reaction
DNA-Directed DNA Polymerase
Genes
Ampicillin Resistance
Bacitracin
Cefuroxime
Norfloxacin
Ceftazidime
Amikacin
Ceftriaxone
Malaysia
Microbial Sensitivity Tests
Streptomycin
Chloramphenicol
Vancomycin

Keywords

  • Antibiotic resistance
  • ERIC-PCR
  • Molecular typing
  • RAPD-PCR
  • Vibrio parahaemolyticus

ASJC Scopus subject areas

  • Biotechnology
  • Physiology
  • Applied Microbiology and Biotechnology

Cite this

@article{86bd3e84121d40a78af8d1e96e07d043,
title = "Antibiotic resistance and molecular typing among cockle (Anadara granosa) strains of Vibrio parahaemolyticus by polymerase chain reaction (PCR)-based analysis",
abstract = "Genomic DNA of Vibrio parahaemolyticus were characterized by antibiotic resistance, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis. These isolates originated from 3 distantly locations of Selangor, Negeri Sembilan and Melaka (East coastal areas), Malaysia. A total of 44 (n = 44) of tentatively V. parahaemolyticus were also examined for the presence of toxR, tdh and trh gene. Of 44 isolates, 37 were positive towards toxR gene; while, none were positive to tdh and trh gene. Antibiotic resistance analysis showed the V. parahaemolyticus isolates were highly resistant to bacitracin (92 {\%}, 34/37) and penicillin (89 {\%}, 33/37) followed by resistance towards ampicillin (68 {\%}, 25/37), cefuroxime (38 {\%}, 14/37), amikacin (6 {\%}, 2/37) and ceftazidime (14 {\%}, 5/37). None of the V. parahaemolyticus isolates were resistant towards chloramphenicol, ciprofloxacin, ceftriaxone, enrofloxacin, norfloxacin, streptomycin and vancomycin. Antibiogram patterns exhibited, 9 patterns and phenotypically less heterogenous when compared to PCR-based techniques using ERIC- and RAPD-PCR. The results of the ERIC- and RAPD-PCR were analyzed using GelCompare software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the V. parahaemolyticus isolates into 6 clusters and 21 single isolates at a similarity level of 80 {\%}. While, RAPD-PCR with primer Gen8 discriminated the V. parahaemolyticus isolates into 11 clusters and 10 single isolates and Gen9 into 8 clusters and 16 single isolates at the same similarity level examined. Results in the presence study demonstrated combination of phenotypically and genotypically methods show a wide heterogeneity among cockle isolates of V. parahaemolyticus.",
keywords = "Antibiotic resistance, ERIC-PCR, Molecular typing, RAPD-PCR, Vibrio parahaemolyticus",
author = "{Abd. Mutalib}, Sahilah and Laila, {R. A S} and Sallehuddin, {H. Mohd} and Osman Hassan and Aminah Abdullah and {Ahmad Azuhairi}, A.",
year = "2014",
month = "2",
doi = "10.1007/s11274-013-1494-y",
language = "English",
volume = "30",
pages = "649--659",
journal = "World Journal of Microbiology and Biotechnology",
issn = "0959-3993",
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TY - JOUR

T1 - Antibiotic resistance and molecular typing among cockle (Anadara granosa) strains of Vibrio parahaemolyticus by polymerase chain reaction (PCR)-based analysis

AU - Abd. Mutalib, Sahilah

AU - Laila, R. A S

AU - Sallehuddin, H. Mohd

AU - Hassan, Osman

AU - Abdullah, Aminah

AU - Ahmad Azuhairi, A.

PY - 2014/2

Y1 - 2014/2

N2 - Genomic DNA of Vibrio parahaemolyticus were characterized by antibiotic resistance, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis. These isolates originated from 3 distantly locations of Selangor, Negeri Sembilan and Melaka (East coastal areas), Malaysia. A total of 44 (n = 44) of tentatively V. parahaemolyticus were also examined for the presence of toxR, tdh and trh gene. Of 44 isolates, 37 were positive towards toxR gene; while, none were positive to tdh and trh gene. Antibiotic resistance analysis showed the V. parahaemolyticus isolates were highly resistant to bacitracin (92 %, 34/37) and penicillin (89 %, 33/37) followed by resistance towards ampicillin (68 %, 25/37), cefuroxime (38 %, 14/37), amikacin (6 %, 2/37) and ceftazidime (14 %, 5/37). None of the V. parahaemolyticus isolates were resistant towards chloramphenicol, ciprofloxacin, ceftriaxone, enrofloxacin, norfloxacin, streptomycin and vancomycin. Antibiogram patterns exhibited, 9 patterns and phenotypically less heterogenous when compared to PCR-based techniques using ERIC- and RAPD-PCR. The results of the ERIC- and RAPD-PCR were analyzed using GelCompare software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the V. parahaemolyticus isolates into 6 clusters and 21 single isolates at a similarity level of 80 %. While, RAPD-PCR with primer Gen8 discriminated the V. parahaemolyticus isolates into 11 clusters and 10 single isolates and Gen9 into 8 clusters and 16 single isolates at the same similarity level examined. Results in the presence study demonstrated combination of phenotypically and genotypically methods show a wide heterogeneity among cockle isolates of V. parahaemolyticus.

AB - Genomic DNA of Vibrio parahaemolyticus were characterized by antibiotic resistance, enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) and random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) analysis. These isolates originated from 3 distantly locations of Selangor, Negeri Sembilan and Melaka (East coastal areas), Malaysia. A total of 44 (n = 44) of tentatively V. parahaemolyticus were also examined for the presence of toxR, tdh and trh gene. Of 44 isolates, 37 were positive towards toxR gene; while, none were positive to tdh and trh gene. Antibiotic resistance analysis showed the V. parahaemolyticus isolates were highly resistant to bacitracin (92 %, 34/37) and penicillin (89 %, 33/37) followed by resistance towards ampicillin (68 %, 25/37), cefuroxime (38 %, 14/37), amikacin (6 %, 2/37) and ceftazidime (14 %, 5/37). None of the V. parahaemolyticus isolates were resistant towards chloramphenicol, ciprofloxacin, ceftriaxone, enrofloxacin, norfloxacin, streptomycin and vancomycin. Antibiogram patterns exhibited, 9 patterns and phenotypically less heterogenous when compared to PCR-based techniques using ERIC- and RAPD-PCR. The results of the ERIC- and RAPD-PCR were analyzed using GelCompare software. ERIC-PCR with primers ERIC1R and ERIC2 discriminated the V. parahaemolyticus isolates into 6 clusters and 21 single isolates at a similarity level of 80 %. While, RAPD-PCR with primer Gen8 discriminated the V. parahaemolyticus isolates into 11 clusters and 10 single isolates and Gen9 into 8 clusters and 16 single isolates at the same similarity level examined. Results in the presence study demonstrated combination of phenotypically and genotypically methods show a wide heterogeneity among cockle isolates of V. parahaemolyticus.

KW - Antibiotic resistance

KW - ERIC-PCR

KW - Molecular typing

KW - RAPD-PCR

KW - Vibrio parahaemolyticus

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U2 - 10.1007/s11274-013-1494-y

DO - 10.1007/s11274-013-1494-y

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JF - World Journal of Microbiology and Biotechnology

SN - 0959-3993

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