Antibiotic contamination in the marine environment forms an emerging threat to marine ecosystems. This study aimed to compare the gut and coelomic microbiota of Stichopus ocellatus with sediments between two coastal districts of Pahang, which potentially conferring as putative biomarkers for sediment pollution monitoring. The composition of the bacteria communities was determined using 16S rRNA V3-region gene amplicon sequencing, while hybrid whole-genome sequencing was employed to analyze the genome of Vibrio parahaemolyticus. The trace elements and antibiotic compositions were access using high-throughput spectrometry. The alpha- and beta-diversity of bacteria in gut and sediment samples from Kuantan differed substantially within (p-value = 0.017604) and between samples (p-value <0.007), respectively. Vibrio genera predominated in Kuantan samples, while Flavobacterium and Synechococcus_E genera predominated in Pekan samples. Vibrio parahaemolyticus revealed the presence of tet(35) and blaCARB₋₃₃ genes that conceived resistance towards tetracycline and beta-lactam antibiotics, respectively, which were detected in sediment and gut samples.

Antimicrobial resistance genes in aquaculture environments have attracted wide interest, since these genes pose a severe threat to human health. This study aimed to explore the possible mechanisms of the ciprofloxacin resistance of Vibrio parahaemolyticus (V. parahaemolytiucs) in aquaculture environments, which may have been affected by the biofertilizer utilization in China. Plasmid-mediate quinolone resistance (PMQR) genes, representative (fluoro)quinolones (FNQs), and ciprofloxacin-resistance isolates in biofertilizer samples were analyzed. The significantly higher abundance of oqxB was alarming. The transferable experiments and Southern blot analysis indicated that oqxB could spread horizontally from biofertilizers to V. parahaemolyticus, and two (16.7%) trans-conjugants harboring oqxB were provided by 12 isolates that successfully produced OqxB. To the best of our knowledge, this study is the first to report PMQR genes dissipation from biofertilizers to V. parahaemolyticus in aquaculture environments. The surveillance, monitoring and control of PMQR genes in biofertilizers are warranted for seafood safety and human health.

Bivalves use anti-oxidative enzyme systems to defend themselves against excessive reactive oxygen species, which are often catalyzed by environmental pollution. As a key member of anti-oxidative enzyme family, catalase plays a crucial role in scavenging the high level of reactive oxygen species to protect organisms against various oxidative stresses. In this study, a catalase homologue was identified from Mytilus coruscus (named McCAT, KX957929). The open reading frame of McCAT was 1844bp with a 5′ untranslated region of 341bp and a 3′ untranslated region of 927bp. The deduced amino acid sequence was 512 residues in length with theoretical pI/MW 8.02/57.91kDa. BLASTn and phylogenetic analyses strongly suggested that it was a member of catalase, also known as CAT family for its conserved catalytic site motif and proximal heme-ligand signature motif. Real-time fluorescence quantitative PCR showed that constitutive expression of McCAT was occurred, with increasing order in mantle, adductor, gill, hemocyte, gonad and hepatopancreas. It was observed that bacterial infection and heavy metals stimulation up-regulated McCAT mRNA expression in hepatopancreas with time-dependent manners. The maximum expression appeared at 8h after pathogenic bacteria injecting, with 15-fold in Vibrio parahemolyticus and 60-fold in Aeromonas hydrophila than that of 0h. The highest point of McCAT mRNA appeared at different times for exposure to heavy metals with copper at day 5 (0.1mg/L 30-fold, 0.5mg/L 15-fold, 1.5mg/L 6-fold) and plumbum at day 3 (3.0mg/L 20-fold). The enzymatic activity analysis found that McCAT activity in the gill of M. coruscus was affected by heavy metals concentration. The results suggested that McCAT plays a significant role in antioxidation and the expression of McCAT can be used as a biomarker for detection of marine environmental pollution.

In this study, we investigated the environmental impacts of scallop culture on two coastal estuaries adjacent the Bohai Sea including developing a quantitative PCR assay to assess the abundance of the bacterial pathogens Escherichia coli and Vibrio parahaemolyticus. Scallop culture resulted in a significant reduction of nitrogen, Chlorophyll a, and phosphorous levels in seawater during summer. The abundance of bacteria including V. parahaemolyticus varied significantly across estuaries and breeding seasons and was influenced by nitrate as well as nutrient ratios (Si/DIN, N/P). Bacterioplankton diversity varied across the two estuaries and seasons, and was dominated by Proteobacteria, Cyanobacteria, Actinobacteria, Bacteroidetes. Overall, this study suggests a significant influence of scallop culture on the ecology of adjacent estuaries and offers a sensitive tool for monitoring scallop contamination.

Vibrio parahaemolyticus and V. vulnificus are the most common pathogens causing seafood-borne illnesses in Korea. This study determines the abundance and antimicrobial resistance of pathogenic Vibrio species in seawater and zooplankton samples from the Geoje Island coast in Korea, which is an important area for coastal fisheries, the fishing industry, and tourism. The two Vibrio species were detected more in mesozooplankton samples than in seawater samples. V. parahaemolyticus isolates showed greater resistance than those of V. vulnificus for antimicrobials. Of V. parahaemolyticus isolates, 93.3% exhibited resistance to three or more antimicrobial agents. Conversely, more than 80% of V. vulnificus isolates showed susceptibility to all antimicrobials examined, with the exception of rifampicin. Our findings show that strong antimicrobial resistance of V. parahaemolyticus in the surveyed area was exposed to conventionally used antibiotics, therefore necessitating proper surveillance programs for the monitoring of antimicrobial resistance patterns in seawater bodies and aquatic animals.

The influence of environmental parameters on the total and pathogenic Vibrio parahaemolyticus seasonal densities in American oysters (Crassostrea virginica) was evaluated for 1year. Harvesting site A yielded the highest mean densities of V. parahaemolyticus tlh+, tdh+/trh−, tdh−/trh+ and tdh+/trh+ during spring season at 2.57, 1.74, 0.36, and −0.40 log10MPN/g, respectively, and tdh+/orf8+ during winter season (0.90 log10MPN/g). V. parahaemolyticus tlh+ densities were associated to salinity (R2=0.372, P<0.022), tdh+/trh+ to turbidity (R2=0.597, P<0.035), and orf8+ to temperature, salinity, and pH (R2=0.964, P<0.001). The exposure to salinity and temperature conditions during winter and spring seasons regulated the dynamics of V. parahaemolyticus harboring potentially pathogenic genotypes within the oyster. The adaptive response of V. parahaemolyticus to seasonal environmental changes may lead to an increase in survival and virulence, threatening the seafood safety and increasing the risk of illness.

The need for quantitative molecular methods is growing in environmental, food, and medical fields but is hindered by low and variable DNA extraction and by co-extraction of PCR inhibitors. DNA extracts from Enterococcus faecium, seawater, and seawater spiked with E. faecium and Vibrio parahaemolyticus were tested by qPCR for target recovery and inhibition. Conventional and novel methods were tested, including Synchronous Coefficient of Drag Alteration (SCODA) and lysis and purification systems used on an automated genetic sensor (the Environmental Sample Processor, ESP). Variable qPCR target recovery and inhibition were measured, significantly affecting target quantification. An aggressive lysis method that utilized chemical, enzymatic, and mechanical disruption enhanced target recovery compared to commercial kit protocols. SCODA purification did not show marked improvement over commercial spin columns. Overall, data suggested a general need to improve sample preparation and to accurately assess and account for DNA recovery and inhibition in qPCR applications.

A presumed Vibrio parahaemolyticus isolate from Chesapeake Bay, Maryland, USA was previously reported to grow on phenanthrene, a polyaromatic hydrocarbon (PAH) found in crude oil. Following the Deepwater Horizon oil spill in the Gulf of Mexico, concerns were raised that PAH-degrading V. parahaemolyticus could increase in abundance, leading to elevated risks of disease derived from shellfish consumption. To assess this possibility, we examined responses to naphthalene and phenanthrene of 17 coastal Louisiana environmental V. parahaemolyticus isolates representing five distinct genotypes. Isolates were obtained immediately after the spill began and after oil had reached the Louisiana coast. None of the isolates grew on or oxidized either substrate and a naphthalene degradation product, 1-naphthol, substantially inhibited growth of some isolates. The use of PAH by V. parahaemolyticus is unusual, and an increase in human health risks due to stimulation of V. parahaemolyticus growth by oil-derived PAH under in situ conditions appears unlikely.

The aquatic environment can represent a reservoir of antimicrobial resistance genes. In the present study, phenotypical, biochemical and molecular techniques were used to screen a collection of marine strains isolated in Italian aquaculture farms to investigate their beta-lactam resistance profiles. The genome of 12 carbapenemase and/or beta-lactamase producing strains was sequenced and a phylogenetic analysis of the beta-lactamases found in their chromosomes was performed. Gene annotation and prediction revealed the presence of blaAₘₚC and blaOXA₋₅₅₋ₗᵢₖₑ in all the Shewanella algae isolates whereas in Vibrio anguillarum and Vibrio parahaemolyticus strains, blaAₘₚC and blaCARB₋₁₉ were found, respectively. Multiple alignments of OXA-55-like and AmpC protein sequences showed different point mutations. Finally, comparisons between enzyme phylogeny and strain clusterization based on sampling sites and dates indicate the diffusion of specific Multi Drug Resistant (MDR) Shewanella algae clones along the Italian Adriatic coast.

In characterization of food borne pathogens from the environment, assessment of virulence, genetic diversity and AMR are essential preludes to formulate preventive strategies and to combat the spread. This study aimed to identify and characterize pathogenic Vibrio parahaemolyticus in the coastal aquaculture farms of Kerala, India. Twenty-seven β-haemolytic V. parahaemolyticus were isolated from 7 out of 40 farms studied. Among the 27 isolates, 15 possessed the tdh gene and 4 had trh. ERIC PCR and PFGE illustrated the presence of pathogenic isolates that shared genetic similarity with clinical strains. One pathogenic isolate was identified to be multidrug resistant (MDR) and 59% exhibited a MAR index of 0.2 or above. Seventy four percent of the pathogenic isolates were ESBL producers and 3.7% of them were carbapenemase producers phenotypically. This asks for adoption of control measures during farming to prevent the transmission of pathogenic V. parahaemolyticus to the environment and food chain.