Inactivation of Human Norovirus GII.4 and Vibrio parahaemolyticus in the Sea Squirt (Halocynthia roretzi) by Floating Electrode-Dielectric Barrier Discharge Plasma.

Human norovirus (HNoV) GII.4 and Vibrio parahaemolyticus may be found in sea squirts. Antimicrobial effects of floating electrode-dielectric barrier discharge (FE-DBD) plasma (5-75 min, N2 1.5 m/s, 1.1 kV, 43 kHz) treatment were examined. HNoV GII.4 decreased by 0.11-1.29 log copy/µL with increasing duration of treatment time, and further by 0.34 log copy/µL when propidium monoazide (PMA) treatment was added to distinguish infectious viruses. The decimal reduction time (D1) of non-PMA and PMA-treated HNoV GII.4 by first-order kinetics were 61.7 (R2 = 0.97) and 58.8 (R2 = 0.92) min, respectively. V. parahaemolyticus decreased by 0.16-1.5 log CFU/g as treatment duration increased. The D1 for V. parahaemolyticus by first-order kinetics was 65.36 (R2 = 0.90) min. Volatile basic nitrogen showed no significant difference from the control until 15 min of FE-DBD plasma treatment, increasing after 30 min. The pH did not differ significantly from the control by 45-60 min, and Hunter color in "L" (lightness), "a" (redness), and "b" (yellowness) values reduced significantly as treatment duration increased. Textures appeared to be individual differences but were not changed by treatment. Therefore, this study suggests that FE-DBD plasma has the potential to serve as a new antimicrobial to foster safer consumption of raw sea squirts.

Application of high hydrostatic pressure for the inactivation of norovirus and quality stability in fresh sea squirt (Halocynthia roretzi).

Sea squirt (Halocynthia roretzi) is considered a potential cause of human norovirus in Korea. This study investigated the effect of high hydrostatic pressure at 100-500 MPa for 5 min at room temperature (23 ± 2 ℃) on the inactivation of murine norovirus-1 (initial inoculum of 6-7 log10 plaque forming units/ml) as a human norovirus surrogate in fresh sea squirt. The effects of high hydrostatic pressure on the Hunter colors and pH were also examined as the main indices of quality. No reductions in murine norovirus-1 titers were observed in sea squirt treated at 100-400 MPa. However, murine norovirus-1 in sea squirt was completely inactivated by 500 MPa of high hydrostatic pressure. Furthermore, the Hunter colors ("L," "a," and "b") and pH values (6.10-6.19) were not significantly (P > 0.05) different between non-high hydrostatic pressure-treated sea squirts and all high hydrostatic pressure-treated sea squirts. Therefore, 500 MPa of high hydrostatic pressure at room temperature may be an optimal treatment for Tunicata meat without altering the food quality (color and pH).

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.

Studies on antimicrobial activity of Poncirus trifoliata ethyl extract fraction against methicillin-resistant Staphylococcus aureus and to elucidate its antibacterial mechanism.

Traditional medicinal plants contain a wide variety of chemicals that have potent antibacterial activity. To find an alternative agent of overcoming the problems of methicillin-resistant Staphylococcus aureus (MRSA), the antibacterial mechanism of Ponciruss trifoliata against MRSA was investigated. Ethyl acetate (EtOAc)-soluble extract of P. trifoliata methanolic extract was evaluated for antibacterial activity using minimum inhibitory concentration (MIC). An EtOAc sub-fraction 08 (EA08) from silica-gel open column chromatography exhibited strong anti-MRSA activity. Apart from the study to isolate single compound from EA08, a synergistic antibacterial effect between the sub-fraction and ß-lactam antibiotics against MRSA was determined. In order to elucidate the antibacterial restoring mechanism of EA08 on MRSA, mRNA expression of mecA gene and production penicillin-binding protein 2a (PBP2a) encoded by mecA gene were monitored. EA 08 showed the strongest antibacterial activity with MIC value of 256 µg ml(-1). MIC of oxacillin against MRSA was dramatically reduced from 512 to 16 µg ml(-1) in combination with 256 µg ml(-1) of EA08. The fractional inhibitory concentration index of oxacillin was measured at 0.53 in combination with EA08 against MRSA, suggesting that EA08-oxacillin combinations exert synergetic effect against MRSA. The analysis of RT-PCR and Western blotting profiles revealed that EA08 inhibited mRNA expression of mecA gene and production PBP2a, which is a key determinant for ß-lactam antibiotic resistance, in a dose-dependent manner. These results indicated that EA08 eventually led to the reduction or inhibition of PBP2a production through translational inhibition in MRSA.

The mechanism of antibacterial activity of phlorofucofuroeckol-A against methicillin-resistant Staphylococcus aureus.

To find more effective ways of overcoming methicillin-resistant Staphylococcus aureus (MRSA), there has been considerable interest in the use of marine-derived constituents as alternatives to control pathogenic microorganisms. In this study, we investigated whether phlorofucofuroeckol-A (PFF) isolated from the edible brown alga Eisenia bicyclis suppressed production or function of penicillin-binding protein 2a (PBP2a). The antimicrobial mode of action of PFF in MRSA was identified by measuring cell membrane integrity and using the time-kill curve method. We attempted to determine the antimicrobial effects of PFF on the expression level of the resistance determinants mecA and its regulatory genes mecI and mecR1 in MRSA by reverse transcriptase polymerase chain reaction. PFF suppressed mecI, mecR1, and mecA gene expression in a dose-dependent manner. In addition, we revealed PFF mediates the suppressive effect of PBP2a expression in MRSA by Western blot analysis. PFF suppressed production of the PBP2a protein, suggesting that PFF probably acts by controlling the methicillin resistance-associated genes involved in the cell wall and production of PBP2a. These results demonstrate that PFF isolated from E. bicyclis significantly suppressed the expression of the methicillin resistance-associated genes and production of PBP2a, which is considered the primary cause of methicillin resistance.