Identification of Aeromonas strains of different origin to the genomic species level

Kaznowski, Adam

doi:10.1046/j.1365-2672.1998.00364.x

Cited by 24 publications

(19 citation statements)

References 24 publications

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“…popoffii (Huys et al . 1997; Kaznowski 1998). DNA–DNA hybridization was not performed in this study, but our strains could be satisfactorily identified to genospecies level by biochemical tests.…”

Section: Discussionmentioning

confidence: 99%

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

González-Rodríguez

Santos

Otero

et al. 2002

Journal of Applied Microbiology

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Aims: Development of a PCR assay for detection of aeromonads carrying the hlyA and ⁄ or aerA genes in fish. Methods and Results: The protocol involves an overnight selective enrichment step in tryptic soy broth yeast extract containing 10 lg ml -1 of ampicillin followed by extraction of DNA and PCR amplification of two haemolysin genes that contribute to the virulence of Aer. hydrophila. This procedure can detect initial populations of 1-10 cfu g )1 within 24 h in artificially contaminated samples. In naturally contaminated fish, both genes were detected in 13 out of 14 fresh fish lots (aeromonads levels between < 1 and 5AE42 log cfu g )1 ) and in 4 out of 16 lots of vacuum-packed cold-smoked fish (aeromonads levels between < 1 and 3AE37 log cfu g )1 ). Before enrichment, dominant species were Aer. hydrophila HG1 (aerA + hlyA + ), Aer. bestiarum HG2 (aerA + hlyA + ) and Aer. caviae HG4 (aerA -hlyA -). After enrichment, Aer. hydrophila HG1 (aerA + hlyA + ) was dominant. Conclusions: Fresh fish and even smoked fish carry hlyA + and ⁄ or aerA + aeromonads that can be detected by PCR within 24 h. Significance and Impact of the Study: The PCR assay described offers considerable potential as a rapid method with specificity, sensitivity and simplicity.

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Section: Discussionmentioning

confidence: 99%

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

González-Rodríguez

Santos

Otero

et al. 2002

Journal of Applied Microbiology

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“…This highlights the need for an improved surveillance system for both the bottled water industry and the municipal water supplies. Since the biochemical characterization is not precise and time-consuming, several proposals have been made about the classification of the aeromonads, including the structural factures, such as the fatty acids methyl esters compositions (FAMES) (Canonica and Pisano, 1985;Canonica and Pisano, 1988) and genetic composition and variability (Soriano, et al, 1997;Kaznowski, 1998;Lee et al, 2002;Miñana-Galbis et al, 2004). The 16S (or small subunit) rDNA sequences have been proven to be a valuable tool in the identification of most Aeromonas species (Martínez-Murcia, 1992), on the members of this genus exhibited very high levels of overall sequence similarity, reaching more than 98% (Figueras et al, 2000).…”

Section: Resultsmentioning

confidence: 99%

The occurrence of Aeromonas spp. in the bottled mineral water, well water and tap water from the municipal supplies

Scoaris

Bizerra

Yamada‐Ogatta

et al. 2008

Braz. arch. biol. technol.

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“…A great deal of confusion and controversy has prevailed over the taxonomic classification of Aeromonas species due to the lack of precise biochemical and/or other phenotypic properties and inadequacy of reliable typing schemes (Kaznowski 1998, Abbott et al 2003. On the basis of 16S rDNA sequencing, all species of the genus Aeromonas are presently included under a distinct family, Aeromonadaceae.…”

Section: Discussionmentioning

confidence: 99%

Multiplex-PCR for simultaneous detection of 3 bacterial fish pathogens, Flavobacterium columnare, Edwardsiella ictaluri, and Aeromonas hydrophila

Panangala

Shoemaker²,

Santen³

et al. 2007

Dis. Aquat. Org.

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A multiplex PCR (m-PCR) method was developed for simultaneous detection of 3 important fish pathogens in warm water aquaculture. The m-PCR to amplify target DNA fragments from Flavobacterium columnare (504 bp), Edwardsiella ictaluri (407 bp) and Aeromonas hydrophila (209 bp) was optimized by adjustment of reaction buffers and a touchdown protocol. The lower detection limit for each of the 3 bacteria was 20 pg of nucleic acid template from each bacteria per m-PCR reaction mixture. The sensitivity threshold for detection of the 3 bacteria in tissues ranged between 3.4 × 10 2 and 2.5 × 10 5 cells g -1 of tissue (channel catfish Ictalurus punctatus Rafinesque). The diagnostic sensitivity and specificity of the m-PCR was evaluated with 10 representative isolates of each of the 3 bacteria and 11 other Gram-negative and 2 Gram-positive bacteria that are taxonomically related or ubiquitous in the aquatic environment. Except for a single species (A. salmonicida subsp. salmonicida), each set of primers specifically amplified the target DNA of the cognate species of bacteria. m-PCR was compared with bacteriological culture for identification of bacteria in experimentally infected fish. The m-PCR appears promising for the rapid, sensitive and simultaneous detection of Flavobacterium columnare, E. ictaluri and A. hydrophila in infected fish compared to the timeconsuming traditional bacteriological culture techniques. KEY WORDS: Multiplex-PCR · Fish · Edwardsiella · Flavobacterium · AeromonasResale or republication not permitted without written consent of the publisher Dis Aquat Org 74: 199-208, 2007 most important bacterial diseases affecting the aquaculture industry in the USA (United States Department of Agriculture 2003). The prevalence and impact of septicemic disease in fish caused by Aeromonas hydrophila (referred to as motile aeromonad septicemia, MSA), is less known. However, A. hydrophila is ubiquitous in aquatic ecosystems (Holmes et al. 1996) and recent studies have shown that catfish latently infected with A. hydrophila (carriers) could shed the organism when co-infected with Edwardsiella ictaluri (Nusbaum & Morrison 2002). Since all 3 species of bacteria (Flavobacterium columnare, E. ictaluri, and A. hydrophila) are ubiquitous in the aquatic environment and fish are reared in extensive pond acreages with high stocking densities (20 000 to 30 000 fish ha -1 ), the occurrence of coinfections or multiple infections in the same host should not be overlooked (Jack et al. 1992). Traditional methods of diagnosing bacterial infections using culture techniques require several days to arrive at a definitive diagnosis, resulting in delayed implementation of control measures and increased potential for spreading of disease. Because PCR can target unique genetic sequences of microorganisms, PCR-based nucleic acid amplification techniques have gained recognition as rapid, sensitive, and specific methods for detection of disease-causing pathogens in various aquaculture species (Vantarakis et al. 2000, Del Cero e...

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Identification of Aeromonas strains of different origin to the genomic species level

Cited by 24 publications

References 24 publications

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

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

The occurrence of Aeromonas spp. in the bottled mineral water, well water and tap water from the municipal supplies

Multiplex-PCR for simultaneous detection of 3 bacterial fish pathogens, Flavobacterium columnare, Edwardsiella ictaluri, and Aeromonas hydrophila

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