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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.

The present study investigated the distribution and antimicrobial susceptibility patterns of Vibrio parahaemolyticus in water samples and aquatic animals (fish and shrimp) from major aquaculture farms along the Korean coast in 2018. V. parahaemolyticus is the most common pathogen causing seafood-borne illness. The strain was detected in 34.7% of all samples tested, and was detected at higher levels during summer to autumn when the water temperature is higher. Although more than 90.0% of V. parahaemolyticus isolates were sensitive to 13 of the 15 antimicrobials tested, which is useful for treating V. parahaemolyticus infectious disease, the isolates exhibited higher resistance to two antibiotics (colistin and ampicillin), which should be excluded as treatment options for these infections. Koreans typically enjoy consuming raw seafood. To reduce the potential human health risk of raw seafood consumption, the prevalence and antimicrobial resistance of V. parahaemolyticus in aquaculture environments should be continuously valuated.

This study investigates the effects of dielectric barrier discharge (DBD) plasma treatment (1.1 kV, 43 kHz, N2 1.5 L/min, 10~60 min) on human norovirus (HuNoV) GII.4 infectivity in fresh oysters. HuNoV viability in oysters was assessed by using propidium monoazide (PMA) as a nucleic acid intercalating dye before performing a real-time reverse transcription&ndash, quantitative polymerase chain reaction (RT-qPCR). Additionally, the impact of the DBD plasma treatment on pH and Hunter colors was assessed. When DBD plasma was treated for 60 min, the HuNoV genomic titer reduction without PMA pretreatment was negligible (&lt, 1 log copy number/&micro, L), whereas when PMA treatment was used, HuNoV titer was reduced to &gt, L in just 30 min. D1 and D2-value of HuNoV infectivity were calculated as 36.5 and 73.0 min of the DBD plasma treatment, respectively, using the first-order kinetics model (R2 = 0.98). The pH and Hunter colors were not significantly different (p &gt, 0.05) between the untreated and DBD-plasma-treated oysters. The results suggest that PMA/RT-qPCR could help distinguish HuNoV infectivity without negatively affecting oyster quality following &gt, 30 min treatment with DBD plasma. Moreover, the inactivation kinetics of nonthermal DBD plasma against HuNoV in fresh oysters might provide basic information for oyster processing and distribution.

From 2008 to 2013, the mussel Mytilus galloprovincialis were collected from a major area of mussel production (Changseon area), which is a designated shellfish-cultivating area for export, located on the southern coast of Korea. The samples were analyzed for mercury (Hg) using a direct Hg analyzer and for other metals, such as cadmium (Cd), lead (Pb), arsenic (As), chromium, copper, nickel, and zinc, using inductively coupled plasma mass spectrometry. The concentrations and bioaccumulation of the heavy met- als were determined, and a potential risk assessment was conducted to evaluate their hazards towards human consumption. The concentration and bioaccumulation ratio of Cd were the highest of the three hazardous metals (Cd, Pb, and Hg). The concentrations of hazardous metals in all samples were within the limits set by Korea and other countries. The estimated dietary intake (EDI) was compared to the provisional tolerable daily intake (PTDI) adopted by the Joint FAO/WHO Expert Committee on Food Additives and the U.S. Environmental Protection Agency. The EDIs of all heavy metals tested for mussel samples ranged from 0.01 to 4.99% of the PTDI; the highest value was measured for As. The hazard index (HI) can be used to assess the risk of heavy metal consump- tion associated with contaminated food. The HI for all samples was far less than 1.0, which indicates that the mussels produced in the Changseon area do not represent an appreciable hazard to humans and are fit for consumption.

In coastal areas that are affected by continuous, seasonal or occasional pollution sources, bivalves accumulate biological contaminants such as pathogenic bacteria. We investigated the effect of natural seawater relaying and electrolyzed seawater depuration on the bacteriological quality of artificially contaminated oysters Crassostrea gigas and short-necked clams Ruditapes philippinarum to suggest an alternative method of shellfish sanitation control.When artificially contaminated oysters and short-necked clams (fecal coliform level 1,700 MPN/100 g) were relayed into a sea area of safe bacteriological water quality, the fecal coliform level dropped to below 110 MPN/100 g after 1 day. The bacteriological quality of oysters and short-necked clams that are contaminated at a fecal coliform level of 1,700 MPN/100 g could be improved, and become appropriate for raw consumption by a single day relay under proper environmental conditions. When artificially contaminated oysters (fecal coliform level 330 MPN/100 g) were depurated with electrolyzed seawater, 94% of fecal coliform was eliminated after 12 h and fecal coliform was undetectable after 24 h. After 24 h depuration with electrolyzed seawater, the fecal coliform level of short-necked clams with initial fecal coliform of 2,400 MPN/100 g was below 20 MPN/100 g. However, the fecal coliform level of short-necked clams with initial fecal coliform of 17,000 MPN/100 g was relatively high, at 790 MPN/100 g, even after 24 h of depuration with electrolyzed seawater, because of the repeated cycle of excretion and accumulation of fecal coliform in shellfish tissue under the closed depuration environment. Such natural seawater relaying and electrolyzed seawater depuration can be restrictively applied to improve or secure the bacteriological quality of oysters and short-necked clams in accordance with safety levels for bivalves for raw consumption.

The influences of inland pollution sources because of rainfall events on the bacteriological water quality in Geoje Bay, a major shellfish production area in Korea, were investigated. The sanitary status of sea water and shellfish after rainfall events was also evaluated. The flow rates of 13 streams around Geoje Bay showed 6 to 7-fold increases after 15 to 21 mm of rainfall. Peak pollution was observed in the Naegan Stream, the Gandeok Stream and the Seojeong Stream. The calculated impact area of inland pollution sources was 3.1 ㎢ immediately after 15 mm of rainfall and expanded to 3.5 ㎢ after 24 hours. These calculations of impacted area matched results from fecal coliform analyses with sea water. The distance between the major pollution source in the bay (the Gandeok Stream) and the station with the worst bacteriological water quality immediately after 15 mm of rainfall, which was below the Korean standard, was 0.8 km in a straight line; this distance increased to 2.0 km after a period of 24 hours. The area impacted by inland pollution sources after a 15 mm rainfall event was wider than after a 21 mm rainfall. Although the flow rate from in-land pollution sources was higher, the concentration of fecal coliform in the discharged water was lower after higher rainfall events. These observations corresponded with the results of fecal coliform analyses with sea water samples. According to the evaluation of the influences of inland pollution sources and fecal coliform analyses on sea water and shellfish samples in Geoje Bay, pollutants from inland sources did not reach the boundary line of the shellfish growing area after rainfall events of 15 or 22 mm. The bacteriological water quality of the shellfish growing area in Geoje Bay met the Korean standard and US NSSP requirements for approved shellfish growing areas.Key words: Geoje Bay, Oyster, Fecal coliform, Shellfish growing area, Inland pollution source

In Hansan Geojeman area, 2,050 ha of shellfish growing area has been designated as shellfish production area for export. The main shellfish species from the designated area is oysters. For the sanitary management of the designated area established in Hansan Geojeman area, bacteriological examination of sea water and shellfish at the sampling stations inside and outside of the designated area were performed from January 2006 to December 2008. The range of fecal coliform of 756 sea water samples at 21 stations located in the designated area were 1,600 MPN/100mL. And the range of geometric mean and the estimated 90th percentile of fecal coliform were 1.8~2.9 and 2.7~15.8 MPN/100mL, respectively. Sanitary conditions of the current designated area in Hansan Geojeman meets the required standards of the Fisheries Product Quality Control and National Shellfish Sanitation Program (NSSP, USA) criteria for the approved area. Also, the sanitary status of the shellfish harvested from the designated area met the Korean Shellfish Sanitation Program (KSSP) fecal coliform criterion (

Antimicrobial resistance patterns of Vibrio species isolated from brackish water in Geoje, Tongyeong and Goseong, Gyeongsangnamdo province into which streams, sewage and leachate all flowed. Only 19 strains (10.7%) of 177 V. parahaemolyticus were susceptible to 15 antimicrobials. 146 strains (69.5%) proved resistant against more than one antimicrobial and 12 strains (6.8%) were multi-drug resistant. The resistance rate of 152 strains were 85.9% against AM and 26.6% against RA, 16.4% against AN, 13.6% against Sand 13.0% against TMP. 86 strains of 129 V. cholerae non-O1 (66.7%) were susceptible to antimicrobials and 31 strains (24.0%) were resistant to more than one antimicrobial and 12 strains (9.3%) were multi-drug resistant. The antimicrobial resistance rate of 129 strains against 15 antimicrobials, with the exception of C, CIP, E and GM, i.e. 11 antimicrobials, was 0.7-16.2%, 16.2% of 129 strains proved resistant against RA and 13.9% against AM, 9.3% against TMP, 7.7% against SXT and 6.9% against TE. 19 of 49 strains of V. mimicus (38.8%) were susceptible to antimicrobials and 31 strains (61.2%) were resistant against more than one antimicrobial; none of the strains were multi-drug resistant. 15 strains of V. mimicus were resistant against only RA, AmC and TE. The resistance rate was 59.2% against RA (highest) 4.1% against AmC and 2.0% against TE.

An amperometric biosensor based on carbon paste electrodes (CPEs) for the determination of urea was constructed by enzyme (urease/GL-DH)-modified method. Urea was hydrolyzed to NH4 + by catalyzing urease onto the enzyme-modified electrode surface in sample solution. In the presence of α -ketoglutarate and reduced nicotinamide adenine dinucleotide(NADH), a liberated NH4 + produce to L-glutamate and NAD + by Lglutamate dehydrogenase (GL-DH). After the chemical reaction was proceeded, the electrochemical reaction was occurred that an excess of the NADH was oxidized to NAD + . The oxidation current of NADH was monitored at +1.10 volt vs. Ag/AgCl. An optimum conditions of biosensor were investigated: The optimum pH range for catalyzed hydrolysis reaction of urea was pH 7.0-7.4. The linear response range and detection limit were 2.0 × 10 −5 ~2.0 × 10 −4 M and 5.0 × 10 −6 M, respectively. Another physiological species did not interfere

From 2009 to 2013, 80 oyster and 16 seawater samples were collected from the southern coast of Korea, including designated shellfish growing areas for export. The concentrations and bioaccumulation of heavy metals were determined, and a potential risk assessment was conducted to evaluate their hazards towards human consumption. The cadmium (Cd) concentration in oysters was the highest of three hazardous metals, including Cd, lead (Pb), and mercury (Hg), however, below the standards set by various countries. The metal bioaccumulation ratio in oysters was relatively high for zinc and Cd but low for Hg, Pb, arsenic, and chromium. The estimated dietary intakes of all heavy metals for oysters accounted for 0.02%-17.75% of provisional tolerable daily intake. The hazard index for all samples was far

A total of 50 raw milk samples from Gyeongnam Province of Korea were examined for the incidence of Listeria monocytogenes between July 1998 and August 1998. L. monocytogenes isolated by biochemical test was confirmed by polymerase chain reaction (PCR) with two sets of primers designed from the invasion-associated protein (iap) gene. After standard PCR with external primers, the amplified DNA was confirmed by a second round of PCR with internal primers (nested PCR). Both the external and internal primers generated 468-bp and 287-bp products, respectively. Only one (G9 strain) of the three suspect samples that tested positive in biochemical tests for L. monocytogenes from 50 raw milk samples was also PCR positive. Following this procedure, PCR-positive G9 strain was confirmed by Southern blot using the 287-bp internal iap probe again. The detection limit of G9 strain by standard PCR assay was as few as 10 2 cells, equivalent to approximately I pg of L. monocytogenes DNA. These PCR assays may be useful for novel detection as well as rapid confirmation for L. monocytogenes from food samples and the field. 50 laits crus ont ete analyses entre juillet et aout 1998. Listeria monocytogenes est isolee et testee par PCR. 2 amorces correspondant au gene iap sont utilisees. 2 PCR sont effectuees. 1 echantillon est teste positif par PCR. La limite de detection par PCR est 10 2 cellules soit 1pg de DNA de L.monocytogenes. Cette methode est sensible et rapide.

From 2011 to 2013, we conducted a full sanitary survey of pollution sources in proximity to a shellfish growing area in the Hansan-Geojeman region in Korea, which includes a designated shellfish growing area. In the sea area, 1152 seawater and 209 oyster samples were collected and examined to evaluate their bacteriological quality. There were 758 potential pollution sources in the drainage area, including 40 sources discharging water in 2013. Fecal coliform (FC) concentrations and impact radii of discharges ranged from 1.8 to 700,000 MPN/100 mL and from 3 to 600 m, respectively; however, the pollutants did not reach the designated area. This demonstrates that the dilution of waste was sufficient such that no significant impact occurred within the designated shellfish growing area. The variation in the FC levels of seawater was closely related to season and rainfall. The FC levels of seawater and oysters from the designated area met the regulation limits set by various countries. No pathogens were found in any oysters. The results of the survey indicate that the oysters produced in this area are apparently safe for raw consumption based on their bacterial quality.