Improving the microbiological safety and quality of aquatic products using nonthermal processing.

Spoilage and deterioration of aquatic products during storage are inevitable, posing significant challenges to their suitability for consumption and the sustainability of the aquatic products supply chain. Research on the nonthermal processing of fruit juices, probiotics, dairy products, and meat has demonstrated positive outcomes in preserving quality. This review examines specific spoilage bacteria species and mechanisms for various aquatic products and discusses the principles, characteristics, and applications of six nonthermal processing methods for bacterial inhibition to maintain microbiological safety and physicochemical quality. The primary spoilage bacteria groups differ among fish, crustaceans, and shellfish based on storage conditions and durations. Four metabolic pathways utilized by spoilage microorganisms-peptides and amino acids, nitrogen compounds, nucleotides, and carbohydrates-are crucial in explaining spoilage. Nonthermal processing techniques, such as ultrahigh pressure, irradiation, magnetic/electric fields, plasma, and ultrasound, can inactivate microorganisms, thereby enhancing microbiological safety, physicochemical quality, and shelf life. Future research may integrate nonthermal processing with other technologies (e.g., modified atmosphere packaging and omics) to elucidate mechanisms of spoilage and improve the storage quality of aquatic products.

Characterization of MLST-99 Salmonella Typhimurium and the monophasic variant I:4,[5],12:i:- isolated from Canadian Atlantic coast shellfish.

Salmonella enterica subsp. enterica Typhimurium and its monophasic variant I 1;4,[5],12:i:- (MVST) are responsible for thousands of reported cases of salmonellosis each year in Canada, and countries worldwide. We investigated S. Typhimurium and MVST isolates recovered from raw shellfish harvested in Atlantic Canada by the Canadian Food Inspection Agency (CFIA) over the past decade, to assess the potential impact of these isolates on human illness and to explore possible routes of shellfish contamination. Whole-genome sequence analysis was performed on 210 isolates of S. Typhimurium and MVST recovered from various food sources, including shellfish. The objective was to identify genetic markers linked to ST-99, a sequence type specifically associated with shellfish, which could explain their high prevalence in shellfish. We also investigated the genetic similarity amongst CFIA ST-99 isolates recovered in different years and geographical locations. Finally, the study aimed to enhance the molecular serotyping of ST-99 isolates, as they are serologically classified as MVST but are frequently misidentified as S. Typhimurium through sequence analysis. To ensure recovery of ST-99 from shellfish was not due to favourable growth kinetics, we measured the growth rates of these isolates relative to other Salmonella and determined that ST-99 did not have a faster growth rate and/or shorter lag phase than other Salmonella evaluated. The CFIA ST-99 isolates from shellfish were highly clonal, with up to 81 high-quality single nucleotide variants amongst isolates. ST-99 isolates both within the CFIA collection and those isolated globally carried numerous unique deletions, insertions and mutations in genes, including some considered important for virulence, such as gene deletions in the type VI secretion system. Interestingly, several of these genetic characteristics appear to be unique to North America. Most notably was a large genomic region showing a high prevalence in genomes from Canadian isolates compared to those from the USA. Although the functions of the majority of the proteins encoded within this region remain unknown, the genes umuC and umuD, known to be protective against UV light damage, were present. While this study did not specifically examine the effects of mutations and insertions, results indicate that these isolates may be adapted to survive in specific environments, such as ocean water, where wild birds and/or animals serve as the natural hosts. Our hypothesis is reinforced by a global phylogenetic analysis, which indicates that isolates obtained from North American shellfish and wild birds are infrequently connected to isolates from human sources. These findings suggest a distinct ecological niche for ST-99, potentially indicating their specialization and adaptation to non-human hosts and environments, such as oceanic habitats.

Identification and validation of core microbes for the formation of the characteristic flavor of fermented oysters (Crassostrea gigas).

Self-fermented oyster homogenates were prepared to investigate core microbes and their correlations with flavor formation mechanisms. Five bacterial and four fungal genera were identified. Correlation analysis showed that Saccharomyces cerevisiae, Kazachstania, and L. pentosus were core species for the flavor of fermented products. Four core microbes were selected for inoculation into homogenates. Twelve key aroma compounds with odor activity values >1 were identified by gas chromatography-mass spectrometry. L. plantarum and S. cerevisiae were beneficial for producing key aroma compounds such as 1-octen-3-ol, (E,Z)-2,6-nonadienal, and heptanal. Fermentation with four microbes resulted in significant increases in contents of Asp, Glu, Lys, inosine monophosphate, and guanosine monophosphate, which provided freshness and sweetness. Fermentation with four microbes resulted in high digestibility, antioxidant abilities, and zinc contents. This study has elucidated the mechanism of flavor formation by microbial action and provides a reference for targeted flavor control in fermented oyster products.

Microencapsulation of ginger essential oil using mung bean protein isolate-chitosan complex coacervates: Application in the preservation of crab meatballs and the prediction of shelf life.

Crab meatballs with more unsaturated fat tend to spoil. Ginger essential oil (GEO) with oxidation resistance was encapsulated into microcapsules (GM) by complex cohesion of mung bean protein isolate (MBPI) and chitosan (CS) in a ratio of 8:1 at pH = 6.4, encapsulation efficiency (EE) and payload (PL) of GM (D50 = 26.16 ± 0.45 µm) with high thermal stability were 78.35 ± 1.02% and 55.43 ± 0.64%. GM (0.6%, w/w) did not interfere with the original flavor of crab meatballs, and lowered values of pH, thiobarbituric acid reactive substances (TBARS) and total bacteria counts (TBC) of the products than those spiked with GEO and the control. The prediction accuracy of the logistic first-order growth kinetic equation in line with TBC (2.84%) was better than that of zero-order and Arrhenius coupled equation based on pH (7.48%) and TBARS (5.94%), but all of them could predict the shelf life of crab meatballs containing GM stored at 4-25 °C.

Association between rainfall and Escherichia coli in live bivalve molluscs harvested in Sardinia, Italy.

Rainfall is generally accepted as one of the most important factors associated with an increased level of E. coli in bivalve molluscs. Performing microbiological risk assessment is relevant to official control authorities to determine the sanitary status of harvesting areas and, therefore, develop monitoring strategies and identify management practices that could be used to improve the quality and safety of the final product. The present study aimed to investigate the impact of rainfall on the content of E. coli in bivalve molluscs farmed in Sardinia (Italy). Enumeration of E. coli was performed according to the Most Probable Number (MPN) method (ISO 16649-3) on 1,920 bivalve samples collected from 7 regional counties between 2018 and 2020. Bivalve molluscs samples included 955 mussels (Mytilus galloprovincialis), 500 oysters (Crassostrea gigas), 325 clams (Ruditapes decussatus), 94 warty venus (Venus verrucosa), and 46 lagoon cockles (Cerastoderma glaucum). Rainfall data were obtained by the Department of Meteorology of the ARPA Sardegna. For each sampling site, GPS coordinates were used to identify gauge stations within catchment areas. Cumulative rain (mm) was recorded 1, 3, 5, 7, and 15 days before sampling, among which the 7-day cumulative rain was the strongest predictor of E. coli counts. Several thresholds of 7-day cumulative rain (from <10 mm up to >300 mm) before sampling were used to estimate the chances of a non-compliant sample (E. coli levels above the limit for sanitary class A; 230 MPN/100 g). The 7-day cumulative rain was positively associated with the chances of non-compliance. When the 7-day cumulative rain before sampling was >300 mm, 80.5 % of the samples were non-compliant, and the odds of a non-compliant sample were 23.6 times higher, as compared to samples harvested when the 7-day cumulative rainfall was <10 mm. Precipitation data could be a useful tool for interpreting anomalous results from official control authorities and reduce the costs that originate from closure of production areas.

Predator-Prey Interactions between Halobacteriovorax and Pathogenic Vibrio parahaemolyticus Strains: Geographical Considerations and Influence of Vibrio Hemolysins.

Halobacteriovorax is a genus of naturally occurring marine predatory bacteria that attack, replicate within, and lyse vibrios and other bacteria. This study evaluated the specificity of four Halobacteriovorax strains against important sequence types (STs) of clinically relevant Vibrio parahaemolyticus, including pandemic strains ST3 and ST36. The Halobacteriovorax bacteria were previously isolated from seawater from the Mid-Atlantic, Gulf of Mexico, and Hawaiian coasts of the United States. Specificity screening was performed using a double agar plaque assay technique on 23 well-characterized and genomically sequenced V. parahaemolyticus strains isolated from infected individuals from widely varying geographic locations within the United States. With few exceptions, results showed that Halobacteriovorax bacteria were excellent predators of the V. parahaemolyticus strains regardless of the origins of the predator or prey. Sequence types and serotypes of V. parahaemolyticus did not influence host specificity, nor did the presence or absence of genes for the thermostable direct hemolysin (TDH) or the TDH-related hemolysin, although faint (cloudy) plaques were present when one or both hemolysins were absent in three of the Vibrio strains. Plaque sizes varied depending on both the Halobacteriovorax and Vibrio strains evaluated, suggesting differences in Halobacteriovorax replication and/or growth rates. The very broad infectivity of Halobacteriovorax toward pathogenic strains of V. parahaemolyticus makes Halobacteriovorax a strong candidate for use in commercial processing applications to enhance the safety of seafoods. IMPORTANCE Vibrio parahaemolyticus is a formidable obstacle to seafood safety. Strains pathogenic to humans are numerous and difficult to control, especially within molluscan shellfish. The pandemic spread of ST3 and ST36 has caused considerable concern, but many other STs are also problematic. The present study demonstrates broad predatory activity of Halobacteriovorax strains obtained along U.S. coastal waters from the Mid-Atlantic, Gulf Coast, and Hawaii toward strains of pathogenic V. parahaemolyticus. This broad activity against clinically relevant V. parahaemolyticus strains suggests a role for Halobacteriovorax in mediating pathogenic V. parahaemolyticus levels in seafoods and their environment as well as the potential application of these predators in the development of new disinfection technologies to reduce pathogenic vibrios in molluscan shellfish and other seafoods.

Using a weight of evidence approach to identify sources of microbiological contamination in a shellfish-growing area with "Restricted" classification.

Shellfish-growing areas in rural catchments are occasionally affected by elevated faecal contamination from diffuse sources and may be subject to frequent harvest closures/classification downgrades. We combined traditional risk management methods based on sanitary surveys and monitoring of Escherichia coli in seawater and shellfish with faecal source tracking, bacterial source apportionment, and hydrometeorological modelling to determine the causes of elevated E. coli concentrations contributing to harvest closures in Papanui Inlet (Aotearoa New Zealand). These multiple lines of evidence were used to inform a weight of evidence assessment of bacterial contamination in the inlet. Ruminant livestock was estimated to contribute 80% of the faecal coliform loading. Concentrations of E. coli in seawater were low (≤ 87 MPN 100 ml-1) whilst concentrations in tuaki/cockles/little neck clams (Austrovenus stutchburyi) occasionally exceeded the "Approved" classification limit (230 MPN 100 g-1). The most frequent positive genetic markers in seawater were the seagull (Catellicoccus marimammalium) (54% of seawater samples), and in shellfish, the bovine and seagull markers (both 12.5% of shellfish samples). Solar radiation was negatively correlated with E. coli in tuaki. We found that the growing area is affected by faecal inputs from animal and, to a lesser extent, human (septic tank discharges) sources which elevate contamination to levels detectable in tuaki but not in seawater, particularly in the summer months. The innovative approach can enhance the management of shellfish-growing areas affected by intermittent contamination and enables more targeted action to reduce pollution to improve shellfish water quality.

Current decontamination challenges and potentially complementary solutions to safeguard the vulnerable seafood industry from recalcitrant human norovirus in live shellfish: Quo Vadis?

Safeguarding the seafood industry is important given its contribution to supporting our growing global population. However, shellfish are filter feeders that bioaccumulate microbial contaminants in their tissue from wastewater discharged into the same coastal growing environments leading to significant human disease outbreaks unless appropriately mitigated. Removal or inactivation of enteric viruses is very challenging particularly as human norovirus (hNoV) binds to specific histo-blood ligands in live oyster tissue that are consumed raw or lightly cooked. The regulatory framework that sets out use of clean seawater and UV disinfection is appropriate for bacterial decontamination at the post-harvest land-based depuration (cleaning) stage. However, additional non-thermal technologies are required to eliminate hNoV in live shellfish (particularly oysters) where published genomic studies report that low-pressure UV has limited effectiveness in inactivating hNoV. The use of the standard genomic detection method (ISO 15, 216-1:2017) is not appropriate for assessing the loss of infectious hNoV in treated live shellfish. The use of surrogate viral infectivity methods appear to offer some insight into the loss of hNoV infectiousness in live shellfish during decontamination. This paper reviews the use of existing and potentially other combinational treatment approaches to enhance the removal or inactivation of enteric viruses in live shellfish. The use of alternative and complementary novel diagnostic approaches to discern viable hNoV are discussed. The effectiveness and virological safety of new affordable hNoV intervention(s) require testing and validating at commercial shellfish production in conjunction with laboratory-based research. Appropriate risk management planning should encompass key stakeholders including local government and the wastewater industry. Gaining a mechanistic understanding of the relationship between hNoV response at molecular and structural levels in individually treated oysters as a unit will inform predictive modeling and appropriate treatment technologies. Global warming of coastal growing environments may introduce additional contaminant challenges (such as invasive species); thus, underscoring need to develop real-time ecosystem monitoring of growing environments to alert shellfish producers to appropriately mitigate these threats.

Quantitative Risk Assessment of Vibrio parahaemolyticus in Shellfish from Retail to Consumption in Coastal Cities of Eastern China.

ABSTRACT: Vibrio parahaemolyticus is the main foodborne pathogen worldwide that causes acute gastroenteritis. A quantitative microbiological risk assessment (QMRA) was conducted to evaluate the health risk associated with V. parahaemolyticus in shellfish in the coastal cities in the eastern part of the People's Republic of China. The QMRA framework was established from shellfish at retail to cooking at home to consumption. The prevalence and level of V. parahaemolyticus in shellfish, cooking methods, storage temperature, time after purchase, shellfish consumption frequency, and consumption amount were analyzed in the exposure assessment. The results of the exposure assessment were introduced into the beta-Poisson dose-response model, and Monte Carlo analysis was used to calculate the risk of gastroenteritis from shellfish consumption. The probability of illness caused by V. parahaemolyticus from shellfish consumption per person per year (Pill,yr) was 3.49E-05. Seasonal differences were noted in the Pill/meal; the maximum was 4.81E-06 in summer and the minimum was 2.27E-07 in winter. The sensitivity analysis revealed that the level of V. parahaemolyticus in shellfish and the amount of shellfish consumed per meal were main factors contributing to illness. This QMRA provided valuable information such as the probability of illness associated with the consumption of shellfish and reference points for prevention strategies and control standards of V. parahaemolyticus in shellfish.

Prevalence, distribution and environmental effects on faecal indicator bacteria and pathogens of concern in commercial shellfish production areas in a subtropical region of a developing country (Santa Catarina, Brazil).

This paper reviews recent literature on the abundance and distribution of faecal indicator bacteria and pathogens in shellfish production areas in the state of Santa Catarina, on the subtropical coast of Brazil. This state supplies > 95% of the national production of shellfish. Microbiological monitoring data were mapped using GIS and the results compared with those from other countries. Coastal human population is the main predictive parameter for faecal bacteria in the production areas. Temporal variations of the bacteria can also be predicted by solar radiation and rainfall. The prevalence of pathogens such as hepatitis A virus, human norovirus, Salmonella spp. and Vibrio spp. does not differ substantially from that in developed countries. The information reported here can be used to inform development of microbiological risk profiles for shellfish production areas.

Trh positive strain of Vibrio parahaemolyticus induce immunity by modulating MAPK pathway: A molecular pathogenic insight in immune-related gene regulation.

Vibrio parahaemolyticus is a zoonotic bacterium that causes infections in shellfish, fish and higher vertebrates as well as in humans. The Tdh and Trh positive strains of V. parahaemolyticus are generally considered as major virulent strains. The pathogenic mechanisms of Trh positive strain of V. parahaemolyticus are poorly understood. Therefore, in the present study Indian Major Carp, Labeo rohita was intraperitoneally challenged with a Trh positive strain of V. parahaemolyticus below lethal dose 50 (LD50) to understand the innate immune response. A significant upregulation in the respiratory burst activity, myeloperoxidase activity and lysozyme activity of serum was observed in the challenged fishes. However, the serum alpha (α) 2-macro globulin activity and antiprotease activity remained unaltered in the infected fish. The relative expression study of some immune-related genes showed that after the experimental challenge the expression of immune-related genes viz., Toll-like receptor (TLR), Nucleotide-binding oligomerization domain (NOD), Interleukin-1ß (IL-ß), Interleukin-6 (IL-6), Tumor necrosis factor α (TNFα), Inducible nitric oxide synthase (iNOS), Complement factor 3a (C3a) and Heat shock proteins 70 (Hsp70) was upregulated during infection. Furthermore, overexpression of nuclear factor kappa light chain enhancer of activated B cells (NF-κß), Myeloid differentiation primary response 88 (MyD88), Mitogen-activated protein kinases (MAPK) and cysteine-aspartic proteases (Casp 1) was also observed after post-infection which clearly indicated that Trh positive V. parahaemolyticus activates MAPK pathway. The present study strengthens the understanding of molecular pathogenesis and provides insights on gene regulation during infection with Trh positive V. parahaemolyticus.

Aquaculture industry prospective from gut microbiome of fish and shellfish: An overview.

The microbiome actually deals with micro-organisms that are associated with indigenous body parts and the entire gut system in all animals, including human beings. These microbes are linked with roles involving hereditary traits, defence against diseases and strengthening overall immunity, which determines the health status of an organism. Considerable efforts have been made to find out the microbiome diversity and their taxonomic identification in finfish and shellfish and its importance has been correlated with various physiological functions and activities. In recent past due to the availability of advanced molecular tools, some efforts have also been made on DNA sequencing of these microbes to understand the environmental impact and other stress factors on their genomic structural profile. There are reports on the use of next-generation sequencing (NGS) technology, including amplicon and shot-gun approaches, and associated bioinformatics tools to count and classify commensal microbiome at the species level. The microbiome present in the whole body, particularly in the gut systems of finfish and shellfish, not only contributes to digestion but also has an impact on nutrition, growth, reproduction, immune system and vulnerability of the host fish to diseases. Therefore, the study of such microbial communities is highly relevant for the development of new and innovative bio-products which will be a vital source to build bio and pharmaceutical industries, including aquaculture. In recent years, attempts have been made to discover the chemical ingredients present in these microbes in the form of biomolecules/bioactive compounds with their functions and usefulness for various health benefits, particularly for the treatment of different types of disorders in animals. Therefore, it has been speculated that microbiomes hold great promise not only as a cure for ailments but also as a preventive measure for the number of infectious diseases. This kind of exploration of new breeds of microbes with their miraculous ingredients will definitely help to accelerate the development of the drugs, pharmaceutical and other biological related industries. Probiotic research and bioinformatics skills will further escalate these opportunities in the sector. In the present review, efforts have been made to collect comprehensive information on the finfish and shellfish microbiome, their diversity and functional properties, relationship with diseases, health status, data on species-specific metagenomics, probiotic research and bioinformatics skills. Further, emphasis has also been made to carry out microbiome research on priority basis not only to keep healthy environment of the fish farming sector but also for the sustainable growth of biological related industries, including aquaculture.

Microbiological Quality and Heavy Metal Concentrations in Slipper Oyster (Crassostrea iredalei) Cultured in Major Growing Areas in Capiz Province, Western Visayas, Philippines: Compliance with International Shellfish Safety and Sanitation Standards.

ABSTRACT: The increasing demand for slipper oyster (Crassostrea iredalei) has propelled farmers to expand oyster cultivation areas in the Philippines, chiefly for local consumption and feasibly for export overseas. As filter feeders, oysters can accumulate pathogens from the surrounding waters, and these pathogens can cause foodborne diseases in consumers. Therefore, oyster farming areas must be monitored for microbiological quality and heavy metal concentrations. In the present study, the microbiological quality of oysters and their growing waters in the major oyster farming areas of the Cogon and Palina Rivers and Cabugao Bay (in Roxas City and the Municipality of Ivisan, respectively, Capiz Province, Western Visayas, Philippines) were examined monthly during the wet (May to October) and dry (November to April) seasons over 12 months. Regardless of the sampling period, high levels of fecal coliforms in the water and Escherichia coli in oysters were found, clearly illustrating that these oyster growing areas would meet only the class B standard under the European Union classification system and would be considered "prohibited" for growing oysters under the U.S. classification system. Although Salmonella was occasionally detected in oysters, Vibrio cholerae was not detected and Vibrio parahaemolyticus was within acceptable limits. The heavy metal concentrations in oyster meat were also determined during the wet (July) and dry (March) seasons. Zinc and copper were the most abundant metals detected, and concentrations of lead, cadmium, mercury, and chromium were below the regulatory limits set by the European Union and the U.S. Food and Drug Administration. These oyster culture areas should be rehabilitated immediately to improve the microbiological quality of the oysters. Oysters harvested from these sites must be depurated or relayed to ensure quality and safety.

Prevalence, typing and antimicrobial resistance of Salmonella isolates from commercial shellfish in the North coast of Morocco.

Salmonellosis is one of the most common foodborne illnesses in the world. The irrational use of antibiotics in medicine and in animal nutrition has greatly favored the emergence and spread of resistant strains of non-typhoid Salmonella. This study aims the determination of the prevalence of Salmonella in bivalve mollusks in Northern Morocco, as well as the molecular typing and antibiotic susceptibility testing of the strains isolated from positive samples. In total, 150 samples from shellfish composed of mussels (Mytilus galloprovincialis), clams (Callista chione and Ruditapes descussatus) and oysters (Magallana gigas). Isolated Salmonella were characterized by Molecular techniques PCR, MLST and MLVA, phylogenetically grouped by MLSA, and susceptibilities were determined for 30 antimicrobial drugs using microdilution method by the BD Phoenix Automated Microbiology System. Prevalence of Salmonella enterica subsp. enterica was 12.67%, grouped in four serovars identified as Chester, Hadar, Typhimurium and Kentucky. Five different MLST STs (sequence types) were detected, ST1954 being the most common, which was mostly found in Chester isolates. Forty-two percent of the isolates showed resistance to more than one antibiotic, especially trimethoprim, sulfa drugs, quinolones and ß-lactam. There was a marked change in the serovars and antimicrobial resistance profiles of the Salmonella isolates in this study compared to those in previous studies.

Molecular characterization and antibiotic resistance of Vibrio parahaemolyticus from Indian oyster and their probable implication in food chain.

Vibrio parahaemolyticus is one of the leading causes of diarrhoea and gastroenteritis in human on consumption of raw or insufficiently cooked seafood. This study was aimed at isolating and characterizing the pathogenic and pandemic V. parahaemolyticus from oysters (n = 90) in coastal parts of West Bengal, India; their antibiotic resistance and potential for involvement in the food chain. During bacteriological culture, typical V. parahaemolyticus colony was recovered in 88.9% samples followed by presumptive identification in 71 (78.9%) samples by characteristic biochemical (K/A) test. All the presumptive isolates (n = 71) were confirmed by species specific Vp-toxR PCR assay. Of these, 10 (14.08%) were tdh+ and none for the trh. Further, 5 (50%) of these tdh+ isolates were found to carry the pandemic potential gene in PGS-PCR assay; however, none in GS-PCR. Majority (80%) of these pathogenic (tdh+) isolates belonged to pandemic serovars (OUT: KUT; OUT: K24; O1: KUT; O1:K25; O10: KUT) and only 20% to non-pandemic serovars (OUT: K15; O9:K17). All the isolates (100%) exhibited resistance to cefpodoxime followed by ampicillin and cefotaxime (90%), ceftizoxime (60%), tetracycline (50%), ceftriaxone (40%), ciprofloxacin and nalidixic acid (10% each). Overall, the study findings suggested that 11.1% (10/90) of commonly marketed oysters in this area were harbouring pathogenic V. parahaemolyticus. Moreover, 5.5% (5/90) of the oyster population were harbouring pandemic strains of this pathogen. Besides, the pathogenic isolates from oysters were exhibiting a considerable genetic relatedness (53 to 70%) to human clinical isolates in PFGE analysis that relates to a substantial public health risk. Further, their multidrug resistance added gravity to the antimicrobial resistance (AMR), a globally growing public health threat and this is a critical area of concern especially during the treatment of foodborne gastroenteritis.

Antibiotic resistance in shellfish and major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea.

Shellfish-growing areas in marine environments are affected by pollutants that mainly originate from land, including streams, domestic wastewater, and the effluents of wastewater treatment plants (WWTPs), which may function as reservoirs of antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs). The objective of this study was to identify the occurrence and distribution of antibiotic resistance at five oyster sampling sites and 11 major inland pollution sources in the drainage basin of Kamak Bay, Republic of Korea. Culture-based methods were used to estimate the diversity and abundance of antibiotic-resistant Escherichia coli strains isolated from oysters and major inland pollution sources. The percentages of ARB and multiple antibiotic resistance index values were significantly high in discharge water from small fishing villages without WWTPs. However, the percentages of antibiotic-resistant E. coli isolates from oysters were low, as there was no impact from major inland pollutants. Fourteen ARGs were also quantified from oysters and major inland pollution sources. Although most ARGs except for quinolones were widely distributed in domestic wastewater discharge and effluent from WWTPs, macrolide resistance genes (ermB and msrA) were detected mainly from oysters in Kamak Bay. This study will aid in tracking the sources of antibiotic contamination in shellfish to determine the correlation between shellfish and inland pollution sources.

Lactic fermentation of cooked, comminuted mussel, Perna canaliculus.

The endogenous microflora of mussels, filter feeders, can include pathogens with resulting food safety concerns. The aim was to develop a cook-then-ferment technology to extend shelf life and safety of a ready-to-eat mussels. Only after cooking to destroy the mussel's endogenous microflora could an edible product be made as determined by pH decline after fermentation and the fate of common pathogens. Perna canaliculus was bought live at retail on many dates. Fermentation was with commercial lactic cultures incubated under vacuum at 30 °C for four days. Using one culture containing Pediococcus pentosaceus and Staphylococcus carnosus as a model, pH typically declined to 4.5 to 4.7, and common pathogens, Staphylococcus aureus, Salmonella and Vibrio parahaemolyticus were absent or reduced to acceptable levels. The fate of Listeria monocytogenes was studied with five cultures. These were variably effective at inhibition with one clear success, Chr Hansen's T-SC-150 containing a specific strain of Lactobacillus sakei, and flavour-generating Staphylococcus carnosus. This culture's efficacy was confirmed with sterile extracts of LAB challenging L. monocytogenes in vitro. This culture was also the most rapid fermenter by pH fall. Cook-then-ferment technology may be applied to other novel foods to minimise a disruptive endogenous microflora.

Influence of climatic factors on the temporal occurrence and distribution of total and pathogenic Vibrio parahaemolyticus in oyster culture environments in Taiwan.

This study evaluated the occurrence and distribution of total and pathogenic V. parahaemolyticus in oyster culture environments in Taiwan. V. parahaemolyticus levels in oysters, seawater, and sediment were quantified using the most probable number (MPN) method combined with a qualitative polymerase chain reaction (PCR). Total V. parahaemolyticus was determined based on the presence or absence of tlh gene, whereas pathogenic V. parahaemolyticus was determined based on the detection of tdh and/or trh gene. The results showed that: 1) V. parahaemolyticus was detected in 93% of the collected samples, 2) the mean concentrations of total V. parahaemolyticus in oysters, seawater, and sediment were 4.1 log MPN/g, 2.1 log MPN/mL, and 4.2 log MPN/g, respectively, and 3) variations in the abundance of V. parahaemolyticus was significantly associated with sea surface temperature (SST). Findings in this study could be used to improve the accuracy of the risk assessment model for V. parahaemolyticus in oysters in Taiwan.

Virulence and antimicrobial resistance potential of Aeromonas spp. associated with shellfish.

Aeromonas spp. are associated with seafood-related outbreaks worldwide. In seafood industry, shellfish play a major role in global seafood production. With this emerging trend of shellfish consumption, shellfish-related bacterial infections are being reported frequently. Aeromonas spp. are natural contaminants found in shellfish. Although 36 species have been identified, some species including Aeromonas hydrophila, Aeromonas caviae and Aeromonas veronii biotype sobria have dragged major attention as foodborne pathogenic bacteria. The ability to elaborate a variety of virulence factors of Aeromonas spp. contributes to the pathogenic activities. Also, emerging antimicrobial resistance in Aeromonas spp. has become a huge challenge in seafood industry. Furthermore, multidrug resistance increases the risk of consumer health. Studies have supplied pieces of evidence about the emerging health risk of Aeromonas spp. isolated from seafood. Therefore, the present review was intended to highlight the prevalence, virulence and antimicrobial resistance of Aeromonas spp. isolated from various types of shellfish.

Population structure, case clusters, and genetic lesions associated with Canadian Salmonella 4,[5],12:i:- isolates.

Monophasic Salmonella 4,[5]:12:i:- are a major public health problem because they are one of the top five Salmonella serotypes isolated from clinical cases globally and because they can carry resistance to multiple antibiotics. A total of 811 Salmonella 4,[5]:12:i:- and S. Typhimurium whole genome sequences (WGS) were generated. The various genetic lesions causing the Salmonella 4,[5]:12:i:- genotype were identified and assessed with regards to their distribution in the population of 811 Salmonella 4,[5]:12:i:- and S. Typhimurium isolates, their geographical and temporal distribution, and their association with non-human sources. Several clades were identified in the population structure, and the largest two were associated almost exclusively with a short prophage insertion and insertion of a mobile element carrying loci encoding antibiotic and mercury resistance. IS26-mediated deletions and fljB point mutants appeared to spread clonally. 'Inconsistent' Salmonella 4,[5]:12:i:- isolates associated with specific, single amino acid changes in fljA and hin were found in a single clade composed of water, shellfish, and avian isolates. Inclusion of isolates from different case clusters identified previously by PFGE validated some of the clusters and invalidated others. Some wgMLST clusters of clinical isolates composed of very closely related isolates contained an isolate(s) with a different genetic lesion, suggesting continuing mobility of the implicated element responsible. Such cases may need to be left out of epidemiological investigations until sufficient numbers of isolates are included that statistical significance of association with sources is not impaired. Non-human sources were frequently found in or near clinical case clusters. Prospective surveillance and WGS of non-human sources and retrospective analysis by WGS of isolates from existing culture collections provides data critical for epidemiological investigations of food- and waterborne outbreaks.