Tetrodotoxin and the Geographic Distribution of the Blue-Lined Octopus Hapalochlaena fasciata on the Korean Coast.

The genus Hapalochlaena, including the blue-lined octopus Hapalochlaena fasciata (H. fasciata), is highly toxic. Venomous, blue-lined octopuses were recently found in Korea, but their toxicity, toxin composition, and distribution remain largely unknown. Here we estimated the geographic distribution of the organisms along the Korean coast and clarified their toxicity. Tetrodotoxin (TTX) was present in all three specimens of H. fasciata examined, although the toxicity varied largely between individuals. The mean TTX concentration in the whole body of the three specimens was 6.5 ± 2.2 µg/g (range 3.3-8.5 µg/g). Among the body parts examined, the salivary glands exhibited the highest concentration (22.4 ± 9.7 µg/g). From 2012 to 2021, 26 individuals were obtained nearly every month from different regions of the Korean coast. A non-fatal case of a blue-lined octopus bite was reported along the Korean coast in June 2015. This is the first report on the widespread distribution of blue-lined octopuses on the Korean coast and TTX detection. The widespread distribution of the TTX-bearing H. fasciata along the Korean coast within the temperate zone indicates that the species may soon become a serious health issue in Korea. The toxicity of this species is also a potentially significant human health risk.

Effect of organic acids on the formation of biogenic amines in fermented anchovy sauce comprising raw anchovy materials with different levels of freshness.

We investigated the effect of different levels of organic acids on the formation of biogenic amines in anchovy fish sauce. Fish sauce samples were prepared with fresh anchovies used immediately after being caught (F), and anchovies left at ambient temperature for 4 h (4 h), 18 h (18 h), or 24 h (24 h). Anchovies from each of the four groups were mixed with salt at a 4:1 ratio and then fermented at ambient temperature for varying periods of time. The F and 4 h anchovies contained higher levels of acetic acid, succinic acid, and lactic acid, and the levels increased during the fermentation process. The histamine content of the fish sauce samples prepared using F anchovies ranged from 44.0 to 9.2 mg/kg at 1 and 24 months of fermentation. The histamine content of fish sauce samples prepared using 4 h, 18 h, and 24 h anchovies was 111.5-37.0 mg/kg (4 h), 780.1-560.3 mg/kg (18 h), and 880.6-675.7 mg/kg (24 h). Our findings indicated that the histamine and other biogenic amine levels were closely associated with the degree of freshness of the raw anchovy material. These findings indicate that lower pH levels resulting from organic acids generated by the fresh raw material can effectively inhibit histamine formation.

Distribution and antimicrobial resistance of Vibrio parahaemolyticus isolated from fish and shrimp aquaculture farms along the Korean coast.

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.

Abundance, antimicrobial resistance, and virulence of pathogenic Vibrio strains from molluscan shellfish farms along the Korean coast.

To reduce the outbreaks caused by the major pathogenic Vibrio species, V. parahaemolyticus, V. vulnificus, and V. cholerae, the distribution, antibiotic resistance, and virulence of these Vibrio strains were monitored in shellfish and seawater along the Korean coast. Among the Vibrio strains, V. parahaemolyticus was the most abundant species; during summer, this strain showed a substantial increase that correlated with the water temperature. Although >99.0% of the Vibrio species isolates were sensitive to seven antimicrobials recommended by the Center for Disease Control and Prevention for the treatment of Vibrio infections, multiple-antibiotic resistance to at least three antimicrobials was found in 14.3% to 50.0% of each Vibrio species. Among V. parahaemolyticus isolates, 14.3% were positive for the trh gene, whereas only 1% was positive for the tdh gene. These results should aid in implementing proper precautions to avoid potential human health risks associated with exposure to pathogenic Vibrio species.

Occurrence, virulence, and antimicrobial resistance of Vibrio parahaemolyticus isolated from bivalve shellfish farms along the southern coast of Korea.

Vibrio parahaemolyticus is the most common pathogen causing seafood-borne illnesses in Korea. The present study evaluated the occurrence, virulence, and antimicrobial resistance of V. parahaemolyticus in seawater and bivalves obtained in 2016 from the southern coast of Korea, an important region for commercial aquaculture industries, especially the Korean raw seafood culture. V. parahaemolyticus was detected in 87 of 160 (54.4%) bivalve samples and in 32 of 130 (24.5%) seawater samples. Especially high levels were detected during summer to early autumn. All the seawater and bivalves contained less than 2 and 5% of the tdh and trh genes of the isolates, respectively, and seawater isolates possessed two fewer genes than the bivalve isolates. Of 23 antimicrobials tested, three agents (ofloxacin, norfloxacin, and trimethoprim/sulfamethoxazole) effectively treated V. parahaemolyticus illness due to the sensitivity of the isolates. The isolates were highly resistant to ampicillin, however, excluding it as a treatment option. More than half of the isolates exhibited resistance to at least three antimicrobials. These findings indicate the importance of an integrated monitoring and surveillance program noting the occurrence, virulence, and antimicrobial resistance patterns of V. parahaemolyticus in various aquatic sources for preventing human health risks from seafood consumption.

Connectivity mapping of angiotensin-PPAR interactions involved in the amelioration of non-alcoholic steatohepatitis by Telmisartan.

Nonalcoholic fatty liver disease (NAFLD) is a global health problem that is associated with various metabolic disorders. Telmisartan is a potential treatment for NAFLD due to its ability to improve insulin sensitivity and decrease hepatic fat accumulation via modulation of PPARγ, and to suppress hepatic fibrosis by blocking angiotensin II receptors. However, the underlying mechanisms of action of telmisartan have yet to be fully elucidated. In the present study, diabetic nonalcoholic steatohepatitis (NASH) mice (STAM mice) received daily administrations of telmisartan for 6 weeks to assess the improvements in NASH. Hepatic transcriptome analyses revealed that the amelioration of NASH likely occurred through the regulation of inflammatory- and fibrosis-related gene responses. An integrated network analysis including transcriptional and non-transcriptional genes regulated by telmisartan showed that the NAFLD pathway is interconnected with the dysregulated RAS-PPAR-NFκB pathways. The downstream targets of PPARα, PPARδ, and RELA in this network significantly overlapped with telmisartan-induced differentially expressed genes (DEGs), which were verified in palmitate-treated Hepa1c1c7 cell line. This transcriptome approach accompanied with cell-based molecular analyses provided the opportunity to understand the fundamental molecular mechanisms underpinning the therapeutic effects of telmisartan, and will contribute to the establishment of a novel pharmacological treatment for NASH patients.

Occurrence and virulence of Vibrio parahaemolyticus isolated from seawater and bivalve shellfish of the Gyeongnam coast, Korea, in 2004-2016.

We determined the abundance and virulence of Vibrio parahaemolyticus in seawater and bivalves from the Gyeongnam coast in Korea, a major area for the seafood industry, during 2004-2016. V. parahaemolyticus is one of the most common pathogen causing seafood-borne illnesses in Korea, and increases during the summer. Its occurrence in seawater and bivalve samples was seasonally dependent, with high levels during the summer to early autumn. There were more strains in the area of sea continually exposed to inland wastewater. Only 5.1% and 3.5% of V. parahaemolyticus isolates from seawater and bivalves, respectively, had the trh gene, and only the bivalve isolates produced the tdh gene at levels below 2%. Continuous monitoring is clearly needed to reduce seafood-borne outbreaks of disease caused by V. parahaemolyticus, and to reveal the occurrence patterns and the presence of toxic genes of the strains in different marine environments.

Total and Methyl Mercury Concentrations in Antarctic Toothfish (Dissostichus mawsoni): Health Risk Assessment.

The concentrations of total mercury (THg) in different organs of the Antarctic toothfish (Dissostichus mawsoni) collected from CCAMLR research blocks in Subarea 88.3 and Division 58.4.1 off the coast of Antarctica were determined. The results revealed THg concentrations of 0.165 ± 0.095 mg/kg (0.023-0.454 mg/kg, wet weight) in the Antarctic toothfish. In muscle, methyl mercury (MeHg) accounted for approximately 40% of the THg. In a comparison analysis, muscle and liver tended to bioaccumulate the highest levels of THg, and both THg and MeHg contents showed correlations with fish length and weight. Compared with international guidelines, fish contained 2.5-6.4% and 4.0-10.3% of the provisional tolerable weekly intake for THg recommended by the Joint FAO/WHO Expert Committee on Food Additives and the tolerable weekly intake for MeHg proposed by the European Food Safety Authority, respectively. These results suggest that consumption of the Antarctic toothfish presents no health risk to humans.

Comparison of bioaccumulation and elimination of Escherichia coli and male-specific bacteriophages by ascidians and bivalves.

Levels of Escherichia coli and male-specific bacteriophages (MSBs) were determined in the filter feeders obtained from retail markets, commercial farms, and wild beds in Korea. The accumulation and elimination of E. coli and MSBs were compared between ascidians and bivalves (oysters and mussels) during relaying and depuration. E. coli concentrations in ascidians from retail markets ranged between < 20 and 460 most probable number/100 g while MSBs were not detected. E. coli levels in bivalves from commercial farms and wild beds were not significantly different but bacterial levels in ascidians were consistently lower. Ascidians exhibited much lower ability than bivalves to accumulate E. coli and MSBs during relaying in a polluted coastal area. This study also shows that an equilibrium was developed between levels of microbes in water and ascidians and shellfish during relaying. E. coli and MSBs in ascidians decreased quickly during depuration in a clean seawater tank. However, after 1 day, E. coli in bivalves decreased by only 1.1-1.6 logs, and the elimination of MSBs was negligible. Therefore, depuration is an effective means to reduce the health risk of contaminated ascidians.

Distribution and antimicrobial susceptibility of Vibrio species associated with zooplankton in coastal area of Korea.

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.

Spatial and seasonal variation of pollution sources in proximity of the Jaranman-Saryangdo area in Korea.

We aimed to compare the spatial and seasonal distributions of fecal coliforms (FCs) and other physiochemical factors in the drainage basin of the Jaranman-Saryangdo area. Among the pollution sources, the mean daily loads and half-circle radii of FCs were the highest in June. However, the pollutants did not reach the boundary line of the designated area due to an existing buffer zone. The value of the FC geometric mean at station 1 was highest in August during periods of heavy rainfall; however, this value was lower than the regulation limit. The highest daily loads of chemical oxygen demand (COD) and chlorophyll-a (Chl-a) in seawater were in the surface layer in August; however, dissolved oxygen (DO) in the bottom water layer was at its lowest in August. This study demonstrated that season and rainfall have significant effects on the FC, COD, DO, and Chl-a concentrations in seawater.

Bacteriological quality evaluation of seawater and oysters from the Hansan-Geojeman area in Korea, 2011-2013: impact of inland pollution sources.

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.

Bacteriological quality evaluation of seawater and oysters from the Jaranman-Saryangdo area, a designated shellfish growing area in Korea: Impact of inland pollution sources.

From 2011 to 2013, we conducted a full sanitary survey of pollution sources in proximity to a designated shellfish growing area in Korea, and their impact on the sea area therein. From this area, 836 seawater samples and 93 oyster samples were examined to evaluate their bacteriological quality. There were 483 potential pollution sources in the drainage area of the Jaranman-Saryangdo area, including 38 sources discharging water. It demonstrates that while many pollution sources have been identified, no significant impact occurred within the designated shellfish growing area. Variations in fecal coliform (FC) levels in seawater were closely related to rainfall. The FC levels of seawater and oysters from the designated area met the regulation limits set by various countries. Our study indicates that the oysters produced in this area are apparently safe for raw consumption based on their bacterial quality.

Distribution of heavy metals in internal organs and tissues of Korean molluscan shellfish and potential risk to human health.

Molluscan shellfish (gastropods and bivalves) were collected from major fish markets on the Korean coast and analyzed for mercury by direct Hg analyzer and for other metals, such as cadmium, lead, chromium, silver, nickel, copper and zinc, using inductively coupled plasma mass spectrometry. Distribution of heavy metals in muscles, internal organs and whole tissues were determined and a potential risk assessment was conducted to evaluate their hazard for human consumption. Heavy metals were accumulated significantly higher (P < 0.05) in internal organs than in muscles for all species. The mean Cd level, which had the highest level of three hazardous metals (Cd, Pb, and Hg) in all internal-organ samples were above the regulatory limit of Korea and the mean level in whole tissue samples of the selected gastropod species, bay scallop and comb pen shell, exceeded the limit (except in a few cases). The sum of the estimated dietary intake of Cd, Pb and Hg for each part of all tested species accounted for 1.59-16.94, 0.02-0.36, and 0.07-0.16% respectively, of the provisional tolerable daily intake adopted by the Joint FAO/WHO Expert Committee on Food Additives. The hazard index for each part of gastropods and bivalves was below 1.0, however, the maximum HI for internal organs of all analysed species was quite high (0.71). These results suggest that consumption of flesh after removing the internal organs of some molluscan shellfish (all gastropod species, bay scallop and comb pen shell) is a suitable way for reducing Cd exposure.

Bioaccumulation of heavy metals in oysters from the southern coast of Korea: assessment of potential risk to human health.

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 <1.0, which indicates that the oysters do not pose an appreciable hazard to humans for the metal pollutants of study.

Distribution of heavy metals in muscles and internal organs of Korean cephalopods and crustaceans: risk assessment for human health.

Samples of seven species of cephalopods and crustaceans were collected from major fish markets on the Korean coast and analyzed for mercury (Hg) using a direct Hg analyzer and for the metals cadmium (Cd), lead (Pb), chromium, silver, nickel, copper, and zinc using inductively coupled plasma mass spectrometry. The distributions of heavy metals in muscles, internal organs, and whole tissues were determined, and a risk assessment was conducted to provide information concerning consumer safety. The heavy metals accumulated to higher levels (P < 0.05) in internal organs than in muscles for all species. The mean concentrations of Cd, which had the highest concentrations of the three hazardous metals (Cd, Pb, and Hg), in all internal organs (except those of blue crab) exceeded the regulatory limits set by Korea and the European Union. The Cd concentrations in all whole tissues of squid and octopus (relatively large cephalopods), red snow crab, and snow crab exceeded the European Union limits. The estimated dietary intake of Cd, Pb, and Hg for each part of all species accounted for 1.73 to 130.57%, 0.03 to 0.39%, and 0.93 to 1.67%, respectively, of the provisional tolerable daily intake adopted by the Joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives; the highest values were found in internal organs. The hazard index (HI) is recognized as a reasonable parameter for assessing the risk of heavy metal consumption associated with contaminated food. Because of the high HI (>1.0) of the internal organs of cephalopods and the maximum HI for whole tissue of 0.424, consumers eating internal organs or whole tissues of cephalopods could be at risk of high heavy metal exposure. Therefore, the internal organs of relatively large cephalopods and crabs (except blue crab) are unfit for consumption. However, consumption of flesh after removing internal organs is a suitable approach for decreasing exposure to harmful metals.

Antibiotic resistance of Vibrio harveyi isolated from seawater in Korea.

Vibrio harveyi is an opportunistic human pathogen that may cause gastroenteritis, severe necrotizing soft-tissue infections, and primary septicemia, with a potentially high rate of lethality. In this study, we isolated and characterized V. harveyi from seawater collected from the West Sea in Korea, including sites located near shellfish farms. For the initial isolation of putative V. harveyi, isolates were incubated on thiosulfate citrate bile salt sucrose agar plates for 24h, followed by selection of greenish colonies. Gram-negative and oxidase-positive colonies were subsequently confirmed by biochemical assays and the API 20E kit test system. Species-specific 16S rRNA and hemolysin genes were used to design V. harveyi-specific PCR primers. From 840 seawater samples, a total of 2 strains of V. harveyi were isolated from shellfish farm seawater. The two isolates were subjected to profiling against 16 antibiotics and found to be resistant to cephalothin, vancomycin, ampicillin, cefepime, cefotetan, and streptomycin.

Contents and risk assessment of heavy metals in marine invertebrates from Korean coastal fish markets.

The concentrations of the heavy metals cadmium (Cd), mercury (Hg), lead (Pb), chromium, silver, nickel, copper, and zinc in the edible portions of 105 marine invertebrates representing 16 mollusk and crustacean species were accurately determined to evaluate their hazard for human consumption. The samples were collected in 2011 from major fish markets on the coast of Korea and analyzed for Hg using a direct Hg analyzer and for other metals using inductively coupled plasma mass spectrometry. Estimated dietary exposure (EDE) was determined, and a risk assessment was made of the heavy metals to provide information concerning consumer safety. The Cd concentrations, which were the highest for the three hazardous metals (Cd, Hg, and Pb), were significantly higher (P < 0.05) in the bivalves and crabs than in the gastropods and cephalopods. However, the concentrations of these metals in all samples were within the regulatory limits set by Korea and other countries. The EDE was compared with the provisional tolerable daily intake (PTDI) adopted by the Joint FAO/WHO Expert Committee on Food Additives or the U.S. Environmental Protection Agency. The EDE of Cd, Hg, and Pb for each class of marine invertebrate were 0.07 to 2.64, 0.01 to 0.43, and 0.001 to 0.16% of the PTDI, respectively. The total EDE of Cd, Hg, and Pb for marine invertebrates accounted for 4.03, 0.96, and 0.21%, respectively, of the PTDI. The EDE of other metals in each class of marine invertebrate was less than 2% of the PTDI. The hazard index is a reasonable parameter for assessing the risk of heavy metal consumption associated with contaminated food. In the present study, the hazard index for all of the species was less than 1.0, which indicates that the intake of heavy metals from consumption of these marine invertebrates does not represent an appreciable hazard to humans.

First detection and seasonal variation of lipophilic toxins okadaic acid, dinophysistoxin-1, and yessotoxin in Korean gastropods.

Okadaic acid (OA), dinophysistoxin-1 (DTX1), pectenotoxin-2, and yessotoxin (YTX) are classes of lipophilic toxins found in marine animals. OA and DTX1 accumulation causes diarrhetic shellfish poisoning, a worldwide public health problem. Diarrhetic shellfish poisoning has not previously been reported in gastropods, which are widely consumed in Korea. Seasonal variation in marine lipophilic toxins in gastropods was investigated using liquid chromatography-tandem mass spectrometry. Eighty specimens of Neptunea cumingii, 65 specimens of Rapana venosa, and 95 specimens of Batillus cornutus were collected at the Tongyeong fish market on the southern coast of Korea between May 2009 and December 2010. OA, DTX1, and YTX were detected in meat and digestive glands in all gastropod species studied. Pectenotoxin-2 was not found in any sample tested. Lipophilic toxins were detected in the digestive glands of gastropods; no lipophilic toxin was detected in the salivary glands of the carnivorous gastropods, N. cumingii and R. venosa. The highest concentrations of OA (21.5 ng/g) and DTX1 (8.4 ng/g) were detected in the digestive glands of R. venosa, and the maximum concentration of YTX (13.7 ng/g) was found in the digestive glands of N. cumingii. The maximum toxicities in gastropod tissues were lower than the European standard for acceptable levels. The concentrations of lipophilic toxins in carnivorous gastropods showed a high degree of seasonal variation; lipophilic toxins in carnivorous gastropods were found predominantly in spring and summer. This is the first report of the occurrence of lipophilic toxins in Korean gastropods.

Geographical and annual variation in lipophilic shellfish toxins from oysters and mussels along the south coast of Korea.

To better understand critical aspects of diarrhetic shellfish poisoning (DSP) occurrence in a chief producing region of bivalves in Korea, the geographical and annual variation of DSP toxins and other lipophilic toxins in mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) were investigated by liquid chromatography-tandem mass spectrometry in an area on the south coast of Korea from 2007 to 2009. The total lipophilic shellfish toxin (LST) levels in bivalves showed geographical and annual variations. LSTs were detected mostly in the hepatopancreas of mussels from Jinhae Bay throughout the entire year, except in November and December of 2007, but were almost undetectable in all samples during the entire year in 2009. The peak DSP toxin (okadaic acid plus dinophysistoxin 1) levels in the hepatopancreas of mussels from Jinhae Bay and the Tongyeong region were 945.3 and 37.6 ng/g, respectively. The DSP toxin content was about 10 times higher in mussels than in oysters collected from the same region. The major toxins in bivalves were okadaic acid and dinophysistoxin 1; however, pectenotoxin 2 or yessotoxin was occasionally detected as a major component. The results of a quantitative analysis of phytoplankton showed that Dinophysis acuminata was the most probable source of the LSTs, with the exception of yessotoxin. When the highest DSP toxin level was measured (945.3 ng/g in the hepatopancreas of mussels from Jinhae Bay), the toxin concentration in whole mussel tissue was calculated to be 114.0 ng/g. The calculated highest DSP toxin level in whole oyster tissue from both regions was 15.0 ng/g. The calculated maximum toxicities in whole mussel and oyster tissues were lower than the regulatory limit (160 to 200 ng/g) in Korea, the European Union, and the United States. Korean oysters (242 samples) and mussels (214 samples) were thus deemed safe for consumption. But because such variation was detected in a relatively small area of the coast, it is possible that at some locations or during a specific period LST levels could exceed the standard and a few consumers could be at risk of experiencing DSP.