Initial numbers of bacteria associated with wild (brown trout and pike) and cultured (rainbow trout) freshwater fish as well as with the water in which they were caught were determined. Subsequently, a total of 979 randomly selected isolates were characterized and identified to the genus level. For all counts performed (aerobes, psychrotrophs, anaerobes, Enterobacteriaceae, and enterococci), no significant differences were observed in water samples, the highest level corresponding to psychrotrophs in pike environments (4.23 X 10(3) CFU/ml). Overall, the skin and intestinal content of brown trout were the most contaminated, while rainbow trout specimens (gills and gut) yielded the lowest numbers. For all bacterial groups, pike gills had the highest numbers. Counts for all of the sampling sites compare well with findings in other temperate geographical environments. Biological characteristics (feeding and skin properties) and the use of antimicrobials in aquaculture might have influenced these results. Motile and nonmotile aerobic gram-negative bacteria together with Enterobacteriaceae accounted for 50 to 70% of the psychrotrophs isolated from water. Micrococcaceae, lactic acid bacteria, Bacillus, and coryneforms were also found. The groups represented in psychrotrophic isolates from the outer surfaces do not reflected those detected in water, so it was common that those organisms recovered in significant numbers from fish were not detected in surrounding habitat of the fish. Motile aeromonads and Carnobacterium were the dominant psychrotrophs in the guts of pike and brown trout, respectively. The intestinal content of reared fish gave a high incidence of Bacillus and coryneforms, while Enterobacteriaceae was absent. Again, rearing practices could have influenced this finding. Listeria monocytogenes was not detected in any of the examined samples. Two strains of Salmonella, which belonged to the same serovar and lysotype, were recovered from pond-water samples taken from one facility on different sampling days. From the gut of a pike specimen and from the pike's environment, two Plesiomonas shigelloides strains of different serovars were recovered. These latter four strains were resistant to a considerable number of antimicrobial compounds (multiple antibiotic resistance indices > 0.2).
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.
Three phenotypic identification systems were employed to identify 106 strains of gram-negative, nonmotile, aerobic bacteria obtained during iced storage of wild (Salmo trutta and Esox lucius) and farmed (Oncorhynchus mykiss) freshwater fish. Using diagnostic tables and computer-assisted identification, the isolates were Psychrobacter (64 strains), Acinetobacter (24 strains), Moraxella (6 strains), Chryseobacterium (5 strains), Myroides odoratus (2 strains), Flavobacterium (1 strain), Empedobacter (1 strain), and unidentified (3 strains). Overall similarities of all strains were determined for 108 characters by numerical analysis (simple matching coefficient of similarity [S] and clustering by unweighted pair group average linkage [UPGMA]). At the 77% similarity level, 92 strains formed nine major clusters (3 or more strains) and four small clusters (2 strains). Cluster 1 (25 isolates divided into two main subclusters) could be assigned to Psychrobacter phenylpyruvicus, clusters 2 and 3 (26 isolates) were designated as Psychrobacter immobilis, and clusters 4 (3 isolates) and 7 (4 isolates) were identified as Psychrobacter urativorans and Psychrobacter spp., respectively. Clusters 5 (five isolates), 6 (three isolates), and 9 (five isolates) were labeled as Acinetobacter spp., Acinetobacter johnsonii, and Acinetobacter lwoffii, respectively. Cluster 8 (12 isolates), with a high resemblance to Thornley's phenon 4 (a heterogeneous group of bacteria isolated from poultry and related to Acinetobacter), remained unnamed. The restriction pattern was identical for strains grouped into clusters 2 and 3 (P. immobilis) but was different for the remaining Psychrobacter isolates. A large proportion of isolates belonging to the family Moraxellaceae were closely related. Psychrobacters and A. johnsonii were present in freshly caught fish and river water. In the latter stages of storage, P. phenylpyruvicus and acinetobacters tended to decrease, whereas P. immobilis increased.
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