An enzyme-linked immunosorbent assay for detection of Vibrio vulnificus biotype 2: development and field studies

Biosca, Elena G.; Marco‐Noales, Ester; Amaro, Carmen; Alcaide, Elena

doi:10.1128/aem.63.2.537-542.1997

Cited by 19 publications

(7 citation statements)

References 41 publications

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“…Survivors and immunized eels could act as carriers. In fact, by using enzyme-linked immunoabsorbent assays (ELISAs), we have confirmed the presence of this biotype/serovar in healthy eels from a fish farm that had previously registered several epizootic outbreaks (12). One of the experimental approaches used to demonstrate that the natural niche of a bacterium is the aquatic ecosystem is to perform a long-term survival study in water microcosms.…”

supporting

confidence: 58%

“…A volume of 10 ml of each microcosm was centrifuged (9,000 ϫ g for 1 h at 4°C), and the cells were resuspended in 1 ml of saline solution (SS; 0.9% NaCl, pH 7.0). Microagglutination and enzyme-linked immunosorbent assay were used as described previously (12) to analyze the antigenicity of starved whole cells. Two antisera were used: one raised against starved cells, and another raised against culturable cells.…”

Section: Methodsmentioning

confidence: 99%

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Effects of Salinity and Temperature on Long-Term Survival of the Eel Pathogen Vibrio vulnificus Biotype 2 (Serovar E)

Marco‐Noales

Biosca

Amaro

1999

Appl Environ Microbiol

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Vibrio vulnificus biotype 2 (serovar E) is a primary eel pathogen. In this study, we performed long-term survival experiments to investigate whether the aquatic ecosystem can be a reservoir for this bacterium. We have used microcosms containing water of different salinities (ranging from 0.3 to 3.8%) maintained at three temperatures (12, 25, and 30°C). Temperature and salinity significantly affected long-term survival: (i) the optimal salinity for survival was 1.5%; (ii) lower salinities reduced survival, although they were nonlethal; and (ii) the optimal temperature for survival was dependent on the salinity (25°C for microcosms at 0.3 and 0.5% and 12°C for microcosms at 1.5 to 3.8%). In the absence of salts, culturability dropped to zero in a few days, without evidence of cellular lysis. Under optimal conditions of salinity and temperature, the bacterium was able to survive in the free-living form for at least 3 years. The presence of a capsule on the bacterial cell seemed to confer an advantage, since the long-term survival rate of opaque variants was significantly higher than that of translucent ones. Long-term-starved cells maintained their infectivity for eels (as determined by both intraperitoneal and immersion challenges) and mice. Examination under the microscope showed that (i) the capsule was maintained, (ii) the cell size decreased, (iii) the rod shape changed to coccuslike along the time of starvation, and (iv) membrane vesicles and extracellular material were occasionally produced. In conclusion, V. vulnificus biotype 2 follows a survival strategy similar to that of biotype 1 of this species in response to starvation conditions in water. Moreover, the aquatic ecosystem is one of its reservoirs.

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supporting

confidence: 58%

Section: Methodsmentioning

confidence: 99%

Effects of Salinity and Temperature on Long-Term Survival of the Eel Pathogen Vibrio vulnificus Biotype 2 (Serovar E)

Marco‐Noales

Biosca

Amaro

1999

Appl Environ Microbiol

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“…The biotype 1 of V. vulnificus and the species V. harveyi , V. splendidus , Aeromonas hydrophila and A. sobria were selected as competitors. These species are abundant in the aquatic environments where the VSE strains must be present (Esteve and Garay, 1991; Biosca, 1994; Arias et al ., 1999), and they are able to grow on the selective media currently employed for the isolation of vibrios (Elliot et al ., 1992; Biosca et al ., 1997b).…”

Section: Introductionmentioning

confidence: 99%

Influence of aquatic microbiota on the survival in water of the human and eel pathogen Vibrio vulnificus serovar E

Marco‐Noales

Biosca

Rojo³

et al. 2004

Environmental Microbiology

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The eel and human pathogen Vibrio vulnificus serovar E (biotype 2) is seldom isolated from natural waters, although it can survive in sterilized artificial seawater microcosms for years. The main objective of the present study was to investigate whether aquatic microbiota can limit its survival and recovery from water samples. A set of preliminary experiments of survival in microcosms containing natural seawater and water from eel farms showed that the persistence of this pathogen was mainly controlled by grazing, and secondarily by bacterial competition. The bacterial competition was further analysed in artificial seawater microcosms co-inoculated with selected virulent serovar E (VSE) strains and potential competitors. Competitors included V. vulnificus biotype 1 isolates and strains of selected species that can grow on the selective media designed for V. vulnificus isolation from water samples. Evidences of bacterial competition that was detrimental for VSE recovery were recorded. Thus, some species produced a deleterious effect on VSE strains under starvation, and others were able to use the resources more efficiently under nutrient input. These results suggest that an overgrowth of more efficient competitor bacteria in conventional media used for isolation of V. vulnificus could mask the recovery of VSE strains and explain the scarcity of reports on the isolation of this human and eel pathogen from natural waters.

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“…Antibodies have proved to be very powerful tools, not only for the detection and quantification of pathogens ( Biosca et al ., 1997 ; Dewey et al ., 1997 ; Karpovich‐Tate et al ., 1998 ), but also for detailed structural and biochemical studies across the spectrum of biological disciplines ( Curtis, 1996; Baluska et al ., 1997 ; Sari et al ., 1998 ; Li et al ., 1998 ). Highly specific and quantitative detection of plant pathogens will be increasingly required for the development of resistant varieties by either conventional breeding or genetic engineering.…”

Section: Introductionmentioning

confidence: 99%

Specific polyclonal antibodies for the obligate plant parasite Polymyxa— a targeted recombinant DNA approach

et al. 2000

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Highly specific rabbit polyclonal antibodies for the obligate sugar-beet root parasite, Polymyxa betae, were produced using a novel recombinant DNA approach. Parasite cDNA was selectively isolated from infected roots, expressed in vitro, and the purified protein used to raise antibodies. This produced clean, precisely targeted antibodies, and allowed for rigorous screening of candidate genes and their products at the molecular level prior to animal immunization. This approach selects for genes whose products are highly expressed by the parasite in planta, and five such candidate genes from Polymyxa betae were identified and cloned. Polyclonal antiserum developed using the product of one such gene was found to react specifically with P. betae in sugar-beet roots and with the closely related Polymyxa graminis in barley roots, and to cross-react with Plasmodiophora brassicae in cabbage roots, without the need for further purification. No cross-reaction was detected with protein extracts from potato roots infected by the plasmodiophoromycete Spongospora subterranea. In all cases, there was no interaction with proteins from host plants, or from other microorganisms found in association with uninoculated sugar-beet, barley, cabbage and potato roots.

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An enzyme-linked immunosorbent assay for detection of Vibrio vulnificus biotype 2: development and field studies

Cited by 19 publications

References 41 publications

Effects of Salinity and Temperature on Long-Term Survival of the Eel Pathogen Vibrio vulnificus Biotype 2 (Serovar E)

Effects of Salinity and Temperature on Long-Term Survival of the Eel Pathogen Vibrio vulnificus Biotype 2 (Serovar E)

Influence of aquatic microbiota on the survival in water of the human and eel pathogen Vibrio vulnificus serovar E

Specific polyclonal antibodies for the obligate plant parasite Polymyxa— a targeted recombinant DNA approach

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