Carbapenems belong to the group of last resort antibiotics in human medicine. Therefore, the emergence of growing numbers of carbapenemase-producing bacteria in food-producing animals or the environment is worrying and an important concern for the public health sector. In the present study, a set of 45 Enterobacteriaceae isolated from German retail seafood (clams and shrimps), sampled in 2016, were investigated by real-time PCR for the presence of carbapenemase-producing bacteria. One Escherichia coli (ST10), isolated from a Venus clam (Ruditapes philippinarum) harvested in the Mediterranean Sea (Italy), contained the carbapenemase gene bla VIM-1 as part of the variable region of a class I integron. Whole-genome sequencing indicated that the integron was embedded in a Tn3-like transposon that also contained the fluoroquinolone resistance gene qnrS1. Additional resistance genes such as the extended-spectrum beta-lactamase bla SHV-12 and the AmpC gene bla ACC-1 were also present in this isolate. Except bla ACC-1, all resistance genes were located on an IncY plasmid. These results confirm previous observations that carbapenemase-producing bacteria have reached the food chain and are of increasing concern for public health.
This study was conducted to determine the prevalence of Vibrio spp. in retail seafood in Berlin, Germany. A total of 160 raw seafood samples from supermarkets and seafood shops, consisting of shrimp ( n = 80) and bivalves ( n = 80), were investigated for the presence of Vibrio spp. using the International Organization for Standardization ISO/TS 21872 method and a multiplex PCR. The overall prevalence of Vibrio spp. in retail seafood was 55% (95% CI: 47.2 to 62.8%). The prevalence of Vibrio spp. in shrimp was slightly higher than in bivalves (57.5 versus 52.5%); however, the difference was not statistically significant. Vibrio alginolyticus was the most prevalent species (35.6%), followed by Vibrio parahaemolyticus (27.5%), Vibrio cholerae (6.3%), and Vibrio vulnificus (0.6%). None of the V. parahaemolyticus ( n = 110) isolates encoded tdh/ trh genes, whereas all V. cholerae isolates ( n = 27) were lacking ctxA. Among the chilled samples ( n = 105), the prevalence of Vibrio spp. in unpacked samples was significantly higher than in packed samples ( P = 0.006). Among the packed samples ( n = 55), no significant difference in the prevalence of Vibrio spp. was observed between chilled or frozen products. The results of this study indicated a high prevalence of Vibrio spp. in retail seafood in Germany; positive samples were detected in all types of seafood investigated. The detection of tdh/ trh-negative V. parahaemolyticus isolates should not be neglected because of previous findings on pathogenic strains lacking these virulence markers. Even though thorough cooking might limit the risk of foodborne illness caused by Vibrio, potential cross-contamination during preparation or consumption of raw and undercooked seafood might represent a risk of Vibrio infections.
Retail seafood in Berlin, Germany, was investigated to detect the prevalence and quantitative load of Enterobacteriaceae that produce extended-spectrum β-lactamase (ESBL) and AmpC β-lactamase (AmpC). A total of 160 raw seafood samples were screened for the presence of these bacteria using MacConkey agar supplemented with 1 mg/L cefotaxime after nonselective enrichment. Isolated species were subsequently identified using matrix-assisted laser desorption-ionization time-of-flight analysis. ESBL and AmpC production was tested by the disk diffusion method, and ESBL and AmpC genes were characterized using real-time and conventional PCR assays with DNA sequencing. Spread plating was used for quantification of ESBL- and AmpC-producing Enterobacteriaceae. Overall, these bacteria were detected in 21.3% of seafood samples (34 of 160 samples) with prevalences of 22.5 and 20% for shrimp and bivalves, respectively. Of the positive samples, 91.2% contained an ESBL- or AmpC-producing Enterobacteriaceae load of <100 CFU/g (lower detection limit), and 8.8% contained 100 to 1,000 CFU/g. Among the 45 Enterobacteriaceae isolates, Klebsiella pneumoniae (13 isolates) and Escherichia coli (12 isolates) were the predominant species. ESBL and AmpC genes were detected in 33 isolates, with the majority of isolates harboring bla (27.3%), bla (21.2%), or bla (21.2%). Our study highlights the hazard associated with seafood containing ESBL- and AmpC-producing Enterobacteriaceae in Germany. Even though the contamination levels were low, the high prevalence of ESBL- and AmpC-producing Enterobacteriaceae in seafood might be of concern to public health because of the potential transmission of these bacteria from seafood to humans through the food chain.
Vibrio parahaemolyticus is a major cause of foodborne diseases and a significant threat to human health worldwide. Most of the infections caused by V. parahaemolyticus are usually associated with the consumption of raw or undercooked seafood. This study was conducted to determine the prevalence, quantitative load, and antimicrobial resistance of V. parahaemolyticus in retail seafood in Hanoi, Vietnam. A total of 120 seafood samples consisting of marine fish (n = 30), oysters (n = 30), shrimp (n = 30), and squid (n = 30) were purchased from different traditional markets in Hanoi between May and October 2020. Isolation of V. parahaemolyticus was based on ISO/TS 21872-1:2017, and the most-probable-number (MPN) method was used for quantification. The disk diffusion method was applied for antimicrobial susceptibility testing. Overall, V. parahaemolyticus was detected in 58.33% of the samples. V. parahaemolyticus was most commonly isolated in shrimp samples, with a prevalence of 86.67%, followed by fish (53.33%), squid (53.33%), and oysters (40%). One V. parahaemolyticus isolate from an oyster carrying the trh gene was detected. Of the positive samples, 27.14% contained V. parahaemolyticus counts of less than 2 log MPN/g, whereas 44.29% ranged from 2 to 4 log MPN/g and 28.57% contained more than 4 log MPN/g. Regarding antimicrobial resistance, 85.71% of V. parahaemolyticus isolates were resistant to at least one antibiotic tested. The highest rate of resistance was observed against ampicillin (81.43%), followed by cefotaxime (11.43%), ceftazidime (11.43%), trimethoprim-sulfamethoxazole (8.57%), and tetracycline (2.86%). The results demonstrate the high prevalence and quantitative load and the antimicrobial resistance of V. parahaemolyticus isolated from seafood sold in the study area. HIGHLIGHTS
High hydrostatic pressure (HHP) has been applied to inactivate spoilage and pathogenic micro-organisms in a variety of food products, including seafood. Vibrio sp. are frequently reported as the main cause of foodborne illness associated with consumption of raw or undercooked seafood particularly shellfish worldwide. To date, data on the inactivation of Vibrio sp. via HHP are still limited and most of the trials only investigated HHP application in oysters and clams. This study demonstrates the efficacy of HHP inactivating Vibrio sp. in both pure culture and mussel homogenates.
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