Development of 16S rRNA targeted PCR methods for the detection and differentiation of Vibrio vulnificus in marine environments

Kim, Myoung Sug; Jeong, Hyun Do

doi:10.1016/s0044-8486(00)00495-6

Cited by 55 publications

(39 citation statements)

References 29 publications

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“…The analysis of 16S rRNA gene gave a quick and accurate identification of bacteria (Genmoto et al, 1996;Kim and Jeong, 2001;Tringe and Hugenholtz, 2008;Tang et al, 2011). Many sets of primers have been designed to amplify different regions of 16S rDNA and have been shown to have different sensitivities and specificities (Greisen et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

Isolation and molecular characterization of some bacterial pathogens in El-Serw fish farm, Egypt

El‐Barbary¹,

Hal²

2016

Egypt. J. of Aquatic Biolo. and Fish.

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The current study was carried out to isolate and identify some of bacteria infecting fish of El-Serw fish farm during the harvest season from December to March 2014-2015. Fish samples (155) were collected from different ponds of farm; 65 Nile tilapia (Oreochromis niloticus), 55 catfish (Clarias gariepinus) and 35 mullet (Liza ramada) were used to isolate bacteria in fish organs (gills, liver and kidney) using different selective media, from the examined fish, bacteria belonging to Pseudomonas, Aeromonas and Enterobacteriaceae were isolated. The group most frequently isolated from fish was Aeromonas, the highest number of putative Aeomonas isolates were obtained from O. niloticus 19 (29%), followed C. gariepinus 12 (21.8%) and L. ramada 5 (14.2%). Whereas putative Pseudomonas sp. showed fewer prevalence 9 (13.8%), 8 (14.5%) and 4 (11.4%) from O. niloticus, C. gariepinus and L. ramada, respectively, while only two colonies were recovered from Yersinia Agar Base. Seven isolates were selected and identified by sequencing of 16S rRNA. The analysis of sequence of 16S rRNA gene using universal primers (RW01 and DG74) resulted in the identification of three bacterial isolates of Pseudomonas fluorescens, one of Pseudomonas putida, two of Aeromonas hydrophila and one of Klebsiella oxytoca. The antibiogram test of these isolates revealed their sensitivity to ciprofloxacin, norfloxacin, and gentamycin.This study concluded that the performance 16S rRNA assay has the prospective to create an important contribution to infection management providing rapid of identification the bacterial pathogens in fish.

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Section: Discussionmentioning

confidence: 99%

Isolation and molecular characterization of some bacterial pathogens in El-Serw fish farm, Egypt

El‐Barbary¹,

Hal²

2016

Egypt. J. of Aquatic Biolo. and Fish.

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“…The analysis of 16S rRNA gene presented fast and accurate identification of the detected bacteria [53][54][55]. Many groups of primers have been designed to amplify various regions of 16S rDNA and have been explained to have different specificities and susceptibility [38].…”

Section: Histopathological Examinationmentioning

confidence: 99%

Phenotypic and Genotypic Characterization of Some Pseudomonas sp. Associated with Burkholderia cepacia Isolated from Various Infected Fishes

El‐Barbary¹,

Hal²

2017

J Aquac Res Development

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“…First, each bacterium was inoculated in 5 mL sterile tryptic soy broth (TSB, Difco) and incubated overnight at 28 °C overnight following the method described by Kim and Jeong (2001). Subsequently, 1.0 mL of the bacterial suspension was transferred into microtube and centrifuged at 7,500 g for 5 min at 4 °C.…”

Section: Dna Isolationmentioning

confidence: 99%

Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak

Ransangan¹,

Zainuri²,

Lal³

et al. 2013

MJM

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Aims: High demand for frog meat in Malaysia especially the American bullfrog (Rana catesbeiana) has promoted intensive farming of the animal. However, the farming of American bullfrog is restricted by the occurrence of diseases. This study reports the first isolation of Elizabethkingia meningoseptica from specimens of American bullfrog that suffer from cataract and 'red-leg' syndrome. Methodology and Result: The pathogen was isolated from eyes and internal organs (liver, kidney and spleen) of the diseased bullfrog specimens. All the bacterial isolates were subjected to phenotypic characterization and antibiotic susceptibility assay, and further identified by using the 16S rDNA sequencing analysis. We designed two pair of specific PCR primers (22-25 mers) which are complimentary to the β-lactamase gene in the reference strain of E. meningoseptica ATCC49470. The result showed all the bacterial isolates shared similar phenotypic characters and antibiotic susceptibility. BLAST analysis of the 16S rDNA sequences indicated that the bacterial isolates had very high sequence homology (100%) with E. meningospetica ATCC49470 and E. meningoseptica isolates from mosquito. The two PCR primers were very specific to E. meningoseptica isolates of this study. Conclusion, significance and impact of study: This is the first isolation and characterization of bacterial pathogen, E. meningoseptica in cultured American bullfrog (Rana catesbeina) that suffered from eye cataract and 'red-leg' syndrome in Sabah, Malaysia. It is suspected that one of the possible transmission routes of the bacterial pathogen could be via mosquito bites. The findings suggest that there is urgent requirement for standard guideline of good farming practice to be adopted in frog farms throughout the country. Such a guideline can help in minimizing economic losses, preventing transmission of the zoonotic bacterial pathogen to farm workers, and sustaining the industry in Malaysia and upgrading frog meat quality for international market.

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Development of 16S rRNA targeted PCR methods for the detection and differentiation of Vibrio vulnificus in marine environments

Cited by 55 publications

References 29 publications

Isolation and molecular characterization of some bacterial pathogens in El-Serw fish farm, Egypt

Isolation and molecular characterization of some bacterial pathogens in El-Serw fish farm, Egypt

Phenotypic and Genotypic Characterization of Some Pseudomonas sp. Associated with Burkholderia cepacia Isolated from Various Infected Fishes

Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak

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