PCR detection of potentially pathogenic aeromonads in raw and cold-smoked freshwater fish

González-Rodríguez, María-Nieves; Santos, Jesús A.; Otero, Andrés; García-López, María-Luisa

doi:10.1046/j.1365-2672.2002.01739.x

Cited by 26 publications

(15 citation statements)

References 34 publications

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“…(2003). PCR detection of the haemolytic genes aer A and hly A was performed in isolates and also in 5 ml of each homogenate after enrichment (González‐Rodríguez et al. 2002a).…”

Section: Methodsmentioning

confidence: 99%

Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain

et al. 2006

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Aims: To survey the presence of indigenous and nonindigenous foodborne bacterial pathogens in displayed prepacked portions of fresh marine fish. Methods and Results: A survey of 50 different samples of fresh marine fish (conger, swordfish, sole, grouper and whiting) was conducted over a period of 5 months. Trays of fillets and steaks were obtained at retail level and tested for foodborne bacterial pathogens. Vibrio cholerae and Salmonella were not detected. Two samples (4%) yielded Vibrio strains carrying a DNA fragment specific for Vibrio parahaemolyticus, but resulted negative to PCR amplification of the virulence‐related tdh gene. Levels of motile Aeromonas ranging from 2·29 to 7·20 log CFU g−1 were found in 31 (62%) samples. All fish portions were positive for the Aeromonas hlyA gene and 38 for both aerA and hlyA genes, which may contribute to diarrhoea‐related virulence. The incidence of Listeria monocytogenes was 10%. Levels of Staphylococcus aureus lower than 2 log CFU g−1 were found in 15 (30%) samples. Numbers of presumptive Clostridium perfringens ranging from 1·82 ± 0·22 to 4·26 ± 1·25 log CFU g−1 were detected in 42 (84%) samples. Edwardsiella tarda was detected in two samples of grouper fillets. Conclusions: Displayed portions of raw fish carried bacteria that can cause foodborne disease. The risk posed by fresh fish when properly cooked is low, but high when destined to be consumed raw, undercooked or very lightly processed. Significance and Impact of the Study: This study revealed that raw fish sold in Spain could be a source of foodborne bacterial pathogens. Improvements in handling and processing are needed to minimize the prevalence of pathogenic bacteria.

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“…(2003). PCR detection of the haemolytic genes aer A and hly A was performed in isolates and also in 5 ml of each homogenate after enrichment (González‐Rodríguez et al. 2002a).…”

Section: Methodsmentioning

confidence: 99%

Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain

et al. 2006

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“…It is also a pore-forming cytolysin able to insert into the cell membrane bilayer causing leakage of cytoplasmic contents. The aerA is a major virulence factor of Aeromonas [20,21]. A weak hemolysin, GCAT has been characterized from A. hydrophila and other aeromonads.…”

Section: Discussionmentioning

confidence: 99%

Detection of toxigenic strains of Aeromonas species in foods by a multiplex PCR assay

2010

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Aeromonas hydrophila and other aeromonads are ubiquitous organisms found in meat, vegetables, drinking water and various other food items. They cause diarrhea and extra-intestinal infections in normal and immunocompromised patients. The aim of the study was to develop a multiplex PCR assay for the detection of virulenceassociated genes of Aeromonas including hemolysin (hlyA), aerolysin (aerA), glycerophospholipid-cholesterol acyl transferase (GCAT) alongwith a 16S rRNA gene. Internal amplifi cation control (IAC), which was coamplifi ed with aerA primers, was also included in this study. The results showed that all cultures of Aeromonas were accurately identifi ed by the assay without showing non-specifi city. A. hydrophila could be detected at a range of 10-50 CFU ml -1 from experimentally spiked fi sh, chicken and milk samples following overnight enrichment in alkaline peptone water supplemented with 10 μg/ml ampicillin (APW-A) by this multiplex PCR (mPCR). When evaluated on a total of 74 naturally occurring food samples, four samples were identifi ed to contain Aeromonas by mPCR. All these results were compared to the conventional culture, isolation and biochemical identifi cation procedures. The high throughput and cost-effective mPCR method developed in this study could provide a powerful tool for detection of pathogenic Aeromonas spp. from food and environmental samples and in addition, the method has advantages in terms of specifi city, sensitivity and ease of use compared to other reported PCR methods and DNA hybridization assays.

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“…Aerolysin, encoded by Aer , is a pore‐forming cytolysin, which can directly inlay into the cell membrane bilayer of host and detroy the integrity of cell membrane, therefore results in cell death. It is also regarded as a heat‐labile β‐haemolysin with phospholipase A and C activities, causing haemolysis, cytotoxicity and enterotoxicity (González‐Rodríguez et al ). Enterotoxins encoded by alt , ast or act cause syndromes, such as haemolysis, cytotoxicity and enterotoxicity, which lead to water‐like diarrhoea, gastroenteritis and death by irreversible binding with host cells (Wang et al ; Sen ; Kingombe et al ).…”

Section: Introductionmentioning

confidence: 99%

A novel multiplex PCR assay for rapid detection of virulent Aeromonas in cultured eels

Xiong

Huang

Guo

et al. 2019

J of Applied Microbiology

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Aims Under intensive and stressful aquaculture conditions, cultured eels are highly susceptible to virulent Aeromonas sp. infections. To rapidly and simultaneously confirm Aeromonas isolate and its virulence, a two‐tube multiplex PCR (mPCR) assay incorporating gyrB gene for genus‐specific recognition and seven major virulence genes for virulence assessment was developed. Methods and Results Eight pairs of primers were designed and divided into two groups—gyrB, ahpA, epr and aerA in tube 1 and alt, act, ast and hlyA in tube 2. The optimized mPCR conditions were the same except for their final concentrations. The specificity of the mPCR was validated by the extracted DNA of 10 Aeromonas and 8 non‐Aeromonas species, or mixed DNA templates. Detection limits were determined to be 200 copies per μl in tube 1 and 20 copies per μl in tube 2. The mPCR reproducibility was tested by both artificial challenge and clinical samples. Conclusions The results showed this two‐tube mPCR assay was rapid, specific, sensitive and reliable. Significance and Impact of the Study To our knowledge, this is the first report to distinguish virulent Aeromonas isolates from nonvirulent ones by seven popular and major virulence genes at the genus‐specific level. And it will be useful for large‐scale screening of virulent Aeromonas sp. in cultured eels.

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PCR detection of potentially pathogenic aeromonads in raw and cold-smoked freshwater fish

Cited by 26 publications

References 34 publications

Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain

Occurrence of foodborne pathogenic bacteria in retail prepackaged portions of marine fish in Spain

Detection of toxigenic strains of Aeromonas species in foods by a multiplex PCR assay

A novel multiplex PCR assay for rapid detection of virulent Aeromonas in cultured eels

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