Aeromonas spp. isolated from ready-to-eat seafood on the Norwegian market: prevalence, putative virulence factors and antimicrobial resistance.

AIMS: We aim to investigate the prevalence, putative virulence factors and antimicrobial resistance of mesophilic Aeromonas isolated from ready-to-eat (RTE) seafood available on the Norwegian market, and to assess the potential risks by consuming RTE seafood to consumers. METHODS AND RESULTS: The prevalence of mesophilic Aeromonas in 148 RTE seafood was investigated and the highest prevalence was found in retail sushi (17%), followed by oysters (10%), fresh salmon loins (10%) and scallops (4%). Among 43 Aeromonas isolates, 75% of them were identified as A. media, 23% as A. salmonicida and 2% as A. bestiarum based on partial gryB gene sequencing. Aeromonas isolates were potentially pathogenic due to the presence of four virulence genes: alt (73%), hylA (22%), aerA (17%) and act (6%). In addition, all isolates were resistant to ampicillin and erythromycin. Most of the isolates (98%) were multidrug resistant. CONCLUSIONS: The occurrence of potentially pathogenic and multidrug-resistant Aeromonas strains in RTE seafood implies a potential risk to consumers. Our finding suggests that RTE seafood could be a potential vehicle for the transfer of virulent and multidrug-resistant Aeromonas. SIGNIFICANCE AND IMPACT OF THE STUDY: To our knowledge, this is the first study to report multiple antibiotic resistance in Aeromonas associated with RTE seafood in Norway.

Human pathogens in marine mammal meat ­ a northern perspective.

Only a few countries worldwide hunt seals and whales commercially. In Norway, hooded and harp seals and minke whales are commercially harvested, and coastal seals (harbour and grey seals) are hunted as game. Marine mammal meat is sold to the public and thus included in general microbiological meat control regulations. Slaughtering and dressing of marine mammals are performed in the open air on deck, and many factors on board sealing or whaling vessels may affect meat quality, such as the ice used for cooling whale meat and the seawater used for cleaning, storage of whale meat in the open air until ambient temperature is reached, and the hygienic conditions of equipment, decks, and other surfaces. Based on existing reports, it appears that meat of seal and whale does not usually represent a microbiological hazard to consumers in Norway, because human disease has not been associated with consumption of such foods. However, as hygienic control on marine mammal meat is ad hoc, mainly based on spot-testing, and addresses very few human pathogens, this conclusion may be premature. Additionally, few data from surveys or systematic quality control screenings have been published. This review examines the occurrence of potential human pathogens in marine mammals, as well as critical points for contamination of meat during the slaughter, dressing, cooling, storage and processing of meat. Some zoonotic agents are of particular relevance as foodborne pathogens, such as Trichinella spp., Toxoplasma gondii, Salmonella and Leptospira spp. In addition, Mycoplasma spp. parapoxvirus and Mycobacterium spp. constitute occupational risks during handling of marine mammals and marine mammal products. Adequate training in hygienic procedures is necessary to minimize the risk of contamination on board, and acquiring further data is essential for obtaining a realistic assessment of the microbiological risk to humans from consuming marine mammal meat.

A multiple-locus variable-number tandem repeat analysis (MLVA) of Listeria monocytogenes isolated from Norwegian salmon-processing factories and from listeriosis patients.

The objective of this study was to characterize Listeria monocytogenes isolated from farmed Atlantic salmon (Salmo salar) and the processing environment in three different Norwegian factories, and compare these to clinical isolates by multiple-locus variable-number tandem repeat analysis (MLVA). The 65 L. monocytogenes isolates obtained gave 15 distinct MLVA profiles. There was great heterogeneity in the distribution of MLVA profiles in factories and within each factory. Nine of the 15 MLVA profiles found in the fish-associated isolates were found to match human profiles. The MLVA profile 07-07-09-10-06 was the most common strain in Norwegian listeriosis patients. L. monocytogenes with this profile has previously been associated with at least two known listeriosis outbreaks in Norway, neither determined to be due to fish consumption. However, since this profile was also found in fish and in the processing environment, fish should be considered as a possible food vehicle during sporadic cases and outbreaks of listeriosis.

An evaluation of the duration of efficacy of emamectin benzoate in the control of Caligus curtus Müller infestations in Atlantic cod, Gadus morhua L.

The duration of efficacy of emamectin benzoate in the control of Caligus curtus infestations in Atlantic cod, Gadus morhua L., was studied following an administration of 50 µg kg⁻¹ for seven consecutive days. No lice were found on medicated fish when challenged 1 week (challenge 1) or 5 weeks (challenge 2) following termination of medication, whereas the mean abundance of lice among the unmedicated fish was 17.9 and 19.3 lice per fish in challenge 1 and 2, respectively. Muscle concentrations of 19.5 ± 8.2 ng g⁻¹ and 3.4 ± 0.9 ng g⁻¹, respectively, and skin concentrations of 23.1 ± 10.8 and 4.2 ± 1.0 ng g⁻¹, respectively, were found 27 and 55 days following the termination of medication. Tissue concentrations and the duration of efficacy indicate a dosing regime for emamectin in cod, similar to the regime used for Atlantic salmon, Salmo salar L.

Consumption of drugs for sea lice infestations in Norwegian fish farms: methods for assessment of treatment patterns and treatment rate.

Sea lice are a major problem in Norwegian fish farms; however, data on drug treatment patterns or treatment rates of sea lice infestations are not available. Such data are important for analysing resistance patterns against drugs used for such infestations. The main objective of the present study was to develop a method to estimate the treatment patterns and treatment rates for drugs used in the treatment against sea lice (Lepeophtheirus salmonis and Caligus elongatus) in farm salmonids by means of national sales statistics. Annual sales figures, as weight of active substances, were obtained from the drug wholesalers and the feed mills. The weight of active drug substances is not useful as a unit of measurement of drug use in an epidemiological context because it does not correct for dosage differences and number of repeat treatments. To correct for these factors, we introduced approved daily dose (ADD(farm fish)) and treatment course-doses(farm fish) kg(-1) live-weight fish. To express the drug treatment patterns, the biomass (in weight) of farm salmonids treated with 1 course of a drug were estimated. When measured as kg active substance, the quantities of drugs for the treatment of sea lice infestations declined by 98% during the study period (1989 to 2002) but this figure increased 5-fold when it was corrected for differences in dosage. To correct for amounts of farm salmonids liable to require treatment we estimated the annual treatment rate, defined as the number of treatments for sea lice infestations per biomass slaughtered Atlantic salmon Salmo salar and rainbow trout Oncorhynchus mykiss. The annual treatment rate increased gradually during the study period; however, it varied considerably (range 0.45 to 1.34, mean 0.90). Before 1995, organophosphates were the most frequently used drugs against sea lice; since then pyrethroids have become the dominating drug group.

Epidemiology of bacterial diseases in Norwegian aquaculture--a description based on antibiotic prescription data for the ten-year period 1991 to 2000.

In Norway, antibacterial drugs for use in farmed fishes have to be prescribed by a veterinarian. Moreover, a national surveillance programme requires that copies of all prescriptions be sent to the Directorate of Fisheries. The prescriptions give information regarding fish farm and locality, weight and species of fish to be medicated, diagnosis, type and amount of drug prescribed, and date. These prescription data for the 10 yr period 1991 to 2000 have been recorded and systematised. A total of 6322 prescriptions issued for 11 fish species were registered; Atlantic salmon represented 87% of the prescriptions, rainbow trout 4.5%, arctic char 0.3%, turbot 3.8%, halibut 2.1%, Atlantic cod 1.2%, and European eel 0.3%. European sea-bass, wolf-fish, coalfish and wrasse represented a total of 0.4% of the prescriptions. Furunculosis was the most frequently given diagnosis in Atlantic salmon, accounting for 79% of all prescriptions for this species. Furunculosis was more frequent during the summer and early autumn, and in the western parts of Norway. Fish weighing more than 1 kg seemed to be the most susceptible. However, furunculosis has almost disappeared in Norwegian aquaculture since 1993. Vibriosis gave cause for antibiotic treatments in almost all fish species, and was the most common diagnosis in rainbow trout, halibut, turbot, cod and European eel. In Atlantic salmon, fish of small and medium size (up to 1 kg) seemed to be more at risk from vibriosis, and outbreaks were more frequent during summer, and in the western counties. Cold-water vibriosis was the second most frequently treated disease in Atlantic salmon, creating severe problems mainly in larger fish, in the northern parts of the country, and during winter and spring. The seasonal distribution was similar for winter ulcer disease, the only disease which seemed to be of increasing importance in Atlantic salmon. Non-specific diagnoses, such as 'bacterial infection' and 'fry disease', were given in a much higher proportion of prescriptions for marine fish species than in prescriptions for salmonids.

Prudent use of antibacterial drugs in Norwegian aquaculture? Surveillance by the use of prescription data.

Antibacterial drug treatment in aquaculture during 1991-1996 was investigated using prescription data provided by the Norwegian Government Fish Inspection and Quality Control Service (NFCS). The majority of prescriptions (n = 5401) were for Atlantic salmon and rainbow trout (salmonids), while 383 prescriptions were for other species. Of the 13 different single substances or combinations prescribed during the study period, only 5 were approved for or had been subjected to clinical trials in salmonids. Of the prescriptions for the salmonids, 99% were for approved drugs or drugs subjected to clinical trials. The major proportion of the antibacterial drugs prescribed for other fish species were drugs which were approved for or which had been subjected to clinical trials in salmonids. In all fish species, the prescribing of antibacterial drugs which were neither approved for nor had been subjected to clinical trials was mainly for fish far below slaughter weight. The prescription data were validated against the drug statistics from the wholesalers and feed mills. It was concluded that the data indeed represented antibacterial drug prescribing in Norwegian aquaculture. The prescribing of antibacterial drugs proved to be almost completely reported to NFCS, which is responsible for the control of drug residues in farmed fish in Norway.

An interlaboratory study to find an alternative to the MPN technique for enumerating Escherichia coli in shellfish.

Nine laboratories in eight countries tested 16 batches of common mussels (Mytilus edulis) over a 32 week period in order to find an alternative to the Most Probable Number (MPN) technique to enumerate E. coli. The alternatives investigated included the 3M Petrifilm system, the Merck Chromocult agar method and a Malthus conductance technique. The Petrifilm was found to be unsuitable and was subsequently dropped from the trial. After 669 analyses, a correlation of 0.83 was observed for log E. coli counts between the MPN and Chromocult methods and there was no significant evidence that either method tended to give higher readings than the other. The MPN was slightly better than the Chromocult method for repeatability but the Chromocult was slightly better for reproducibility. However, the observed differences are probably too small to be of practical importance. On the basis of these data therefore, the two methods appear equally suitable for E. coli enumeration in shellfish. There were poor correlations between these methods and the Malthus technique. A small but significant number of samples tested positive on the Malthus instrument but were recorded negative on the MPN and Chromocult tests. Subsequent analysis positively identified E. coli from these Malthus assays. After statistical analysis, errors were noted in both the MPN and Chromocult methods but it was found that there would be no statistical differences if the Chromocult agar were used as an alternative to the MPN technique.

Fate and microbiological effects of furazolidone in a marine aquaculture sediment.

Furazolidone is used in the treatment of bacterial diseases in farmed fish. During application a large proportion of the administered drug reaches the environment directly or via feces. The persistence and metabolism of furazolidone in sediment from a Norwegian salmon farm is described. Furazolidone, in contrast to oxytetracycline and oxolinic acid, is actively metabolized by microorganisms in the sediment. The main metabolite is 3-(4-cyano-2-oxobutylidene-amino)-2-oxazolidone. This is a well known metabolite of the degradation of furazolidone in mammals, fish and Escherichia coli. 3-(4-Cyano-2-oxobutylideneamino)-2- oxazolidone had no detectable antibacterial activity. The half-life of furazolidone in the sediment at 4 degrees C was calculated to be 18 h.