Commercial phenotypic tests (API 20E) in diagnosis of fish bacteria: a review

Popović, Natalija Topić; Čož‐Rakovac, Rozelindra; Strunjak‐Perović, Ivanc̆ica

doi:10.17221/2058-vetmed

Cited by 51 publications

(38 citation statements)

References 26 publications

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“…In our study we confirmed those findings because only about 3 different isolates per fish sample were proved (Table 1). The insufficiency of commercial systems, when testing environmental samples, has been discussed by several authors, and was reviewed by Popovic et al (2007). As apparent, our study does not support the mentioned data and we have demonstrated that both used identification kits have been applicable for reliable identification of enteric bacteria from the normal intestinal bacterial flora in seawater fish.…”

Section: Discussioncontrasting

confidence: 73%

Composition of cultivable enteric bacteria from the intestine of Antarctic fish (family Nototheniidae)

Sedláček¹,

Staňková²,

Švec³

2016

CZECH J ANIM SCI

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ABSTRACT:In this study, the intestinal bacteria of wild Antarctic fish (family Nototheniidae) were examined using traditional culture-based techniques. Bacterial flora of the gut contents of four marine fish species (Notothenia coriiceps, Trematomus bernacchii, Trematomus hansoni, and Trematomus newnesi) was investigated to describe the allochthonous bacteria inhabiting these Nototheniidae fish. A set of 43 fresh and healthy fish was analyzed and intestinal bacteria were retrieved using the dilution plate technique on selective media Endo agar and XLDA agar. A total of 133 different bacterial isolates were obtained and initially characterized by key phenotypical tests. Notothenoid fish gut microbiota showed low species diversity of isolates and intestines were inhabited by 3 different isolates per fish on average. The bacterial colonization of the intestine content of Trematomus newnesi was poor. Curiously, the Gram-negative non-fermenters, including Pseudomonas sp., Vibrio sp., and Alcaligenes-like isolates represented the majority (59%) of intestine isolates grown on the used selective media for enteric bacteria. Based on preliminary identification, only 54 isolates (41%) were tentatively determined as enteric bacteria. The identification of 54 isolates of Gram-negative fermenting rods to the species level was achieved using biochemical characterization by commercial kits ENTEROtest 24 and Biolog GN2 MicroPlate. Results showed that Enterobacter cloacae phenon and Aeromonas hydrophila were predominant bacterial species in the free-living fish intestine from the group of fermenting Gram-negative rods.

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Section: Discussioncontrasting

confidence: 73%

Composition of cultivable enteric bacteria from the intestine of Antarctic fish (family Nototheniidae)

Sedláček¹,

Staňková²,

Švec³

2016

CZECH J ANIM SCI

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“…The use of API 20E as a diagnostic tool for aquatic bacteria has been debated for decades, as reviewed by Popovic et al (2007). According to Alsina & Blanch (1994), this could be a useful tool when adjustments or modifications to the manufacturer manual are being made and as a supplement to other methods of identification.…”

Section: Discussionmentioning

confidence: 99%

Screening and characterisation of potentially pathogenic bacteria associated with Atlantic codGadus morhualarvae: bath challenge trials using a multidish system

Sandlund

Bergh²

2008

Dis. Aquat. Org.

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In intensive aquaculture systems, high concentrations of nutrients and high densities of fish larvae provide favorable conditions for opportunistic pathogenic bacteria to flourish. We screened potentially pathogenic bacterial strains isolated from moribund Atlantic cod Gadus morhua larvae, pollack Pollachius pollachius, coalfish Pollachius virens, Atlantic halibut Hippoglossus hippoglossus, rotifers, algae and water samples from different hatcheries. Three identical challenge experiments tested a total of 53 strains. A multidish system was used: cod eggs were placed in single wells, together with 2 ml of sterile seawater, and exposed to the bacterial cultures. Final bacterial concentrations in the wells were 10 6 and 10 4 CFU ml -1. Eggs and larvae not exposed to bacteria were used as unchallenged controls. Challenged controls were exposed to Vibrio anguillarum strain 610. Eggs were challenged approximately 48 h prior to hatching and mortality was recorded daily throughout the yolk-sac period. In spite of the high challenge dose of 10 6 CFU ml -1, only 5 bacterial strains tested caused higher mortality than the unchallenged controls. Four of these strains were identified by 16S rDNA and gyrase B gene (GyrB) sequencing as resembling V. anguillarum and 1 strain resembled Carnobacterium sp. Most of the larvae exposed to these strains died within 10 d of challenge. Serotyping of the strains resembling V. anguillarum gave inconclusive results. This indicates differences in serology compared to the serotypes O1, O2 and O3, associated with disease. Three bacterial strains seemed to have a slower infection rate, indicating a longer incubation period. The remaining 45 strains did not seem to have a negative effect on larval survival, suggesting that these are not primary pathogens.

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“…2) according to the degrees of association (Donohue et al, 2006;Eddabra et al, 2012). Although designed primarily for clinical specimens, API 20E has found its way in identification of environmental isolates, particularly fish bacterial isolates (Topic Popovic et al, 2007). However, MALDI TOF MS was found a vastly superior alternative to API 20E in identification and classification of aeromonads from water, sludge and fish samples, capable for differentiation of closely related Aeromonas spp.…”

Section: Methodological Differences In Strain Discriminationmentioning

confidence: 99%

“…The API 20E (bioMerieux, Marcy l'Etoile, France) tests were performed according to the manufacturer's instructions with a few alterations in order to adapt the system to the bacteria of freshwater fish: the incubation time was increased to 48-72 h; the incubation temperature was lowered to 22°C; only the fermentation of sugars was allowed by sealing the cups with sterile mineral oil in the carbohydrate tests (Topic Popovic et al, 2007). A seven-digit profile was obtained for every tested isolate and identified by the Apiweb software.…”

Section: Microbiological Analysesmentioning

confidence: 99%

Detection and diversity of aeromonads from treated wastewater and fish inhabiting effluent and downstream waters

Popović

Kazazić

Strunjak‐Perović

et al. 2015

Ecotoxicology and Environmental Safety

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a b s t r a c tA two-season investigation of the wastewater treatment plant (WWTP) effluent, of related waters, sludge and fish across a wide area and 11 stations, with emphasis on Aeromonas spp. was conducted. Aeromonas veronii was the prevailing aeromonad isolated by MALDI TOF MS in the summer period. A rise of Aeromonas hydrophila was observed in summer in raw sewage, treated wastewater and effluent-carrying canal. The ratio of aeromonad species retrieved from fish tissues did not correspond with the water and sludge findings, as in spring in the effluent-carrying canal fish carried Aeromonas salmonicida ssp. salmonicida and Aeromonas bestiarum, while in summer mainly A. veronii and Acinetobacter johnsonii were isolated from fish tissues in the same location. No correlation was established between fecal coliforms/ enterococci and aeromonad occurrence. All retrieved Aeromonas species demonstrated a distinct spectral pattern, with peaks showing unique mass distribution ranging from 4000 to 10,000 Da. Hierarchical clustering separated aeromonads of all isolated species and clustered closely related strains together. Resistance was determined towards amoxicillin, and frequently towards sulfamethoxazole and erythromycin. In summer, a high proportion of water and sludge Aeromonas species demonstrated multiple resistance patterns towards five or more antimicrobials. The quinolone resistance of water aeromonads was mostly related to A. veronii. There are potential health concerns regarding aeromonad exposure amongst recreational fishermen who come into contact with fish inhabiting waters downstream from the WWTP, and WWTP workers who are occupationally exposed to wastewaters and their aerosols.

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Commercial phenotypic tests (API 20E) in diagnosis of fish bacteria: a review

Cited by 51 publications

References 26 publications

Composition of cultivable enteric bacteria from the intestine of Antarctic fish (family Nototheniidae)

Composition of cultivable enteric bacteria from the intestine of Antarctic fish (family Nototheniidae)

Screening and characterisation of potentially pathogenic bacteria associated with Atlantic codGadus morhualarvae: bath challenge trials using a multidish system

Detection and diversity of aeromonads from treated wastewater and fish inhabiting effluent and downstream waters

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