• Journal of fish pathology
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• v.34 no.2
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• pp.185-199
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• 2021

Vibriosis is an important septicemic bacterial disease that affects a variety of commercial fish species, including cultured Dicentrarchus labrax. Nanotechnology has become an important modern tool for fish diseases prevention. Furthermore, nanomaterials have the ability to prevent and treat fish diseases. The current study was aimed to identify the causative agent of massive mortality of D. labrax commercial farm in Alexandria, Egypt. Experimental infection and the median lethal dose (LD50) of pathogenic isolate were assessed. Also, the effect of ginger nanoparticles (GNPs) and Sacchromyces cerevisiae as feed additives for prevention of vibriosis in D. labrax was carried out. Similarly, the tissue immunstimulant genes, IL-1β and TLR2 were measured in the spleen of feeding groups. The clinical signs of naturally diseased D. labrax showed corneal opacity and paleness of gills with excessive mucous secretion. The post-mortem abnormalities were severe hemorrhage and adhesion of internal organs. After bacteriological isolation and identification, the causative agent of mortality in the current study was Vibrio alginolyticus. The LD₅₀ of V. alginolyticus was 1.5×10^5.4 CFU/ml. The experimentally infected D. labrax showed ulceration, exophthalmia and skin hemorrhages. The post-mortem findings of the experimentally infected D. labrax revealed internal hemorrhage, spleen darkness and paleness of liver. There is no mortality and 100% RPS in groups fed GNPs then injected with V. alginolyticus, in those fed a combination of GNPs and S. cerevisiae and a group fed normal diet then injected with physiological saline (control negative), respectively. Contrarily, there was 10% mortality and 87.5 RPS in the group fed S. cerevisae then injected with V. alginolyticus. On the other hand, the control positive group showed 79% mortality. The spleen IL-1β and TLR2 immunostimulant genes were significantly increased in groups of fish fed GNNP, S. cerevisiae and a combination of GNPs and S. cerevisiae, respectively compared to control group. The highest stimulation of those immunostimulant genes was found in the group fed a combination of GNPs and S. cerevisiae, while fish fed S. cerevisiae had the lowest level. Dietary combination of GNPs and S. cerevisiae was shown to be efficient in preventing of vibriosis, with greatest stimulation of spleen IL-1β and TLR2 immunostimulant genes.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.6
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• pp.896-903
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• 2021

For the safety assessment of microbiological and chemical hazards in raw short-neck clam Ruditapes philippinarum distributed in the Yeongnam and Honam areas during the spring season, the contamination levels of total viable bacteria, coliforms, Escherichia coli, and nine pathogenic bacteria (Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens, Listeria monocytogenes, enterohemorrhagic Escherichia coli, Yersinia enterocolitica, Bacillus cereus, Campylobacter jejuni) as microbiological hazards, and heavy metals (lead, cadmium, total mercury), benzopyrene, shellfish poison (paralytic, diarrhetic, amnesic), and radioactivity (¹³¹I, ¹³⁴Cs+¹³⁷Cs) were also analyzed in 15 samples based on the methods of the Korean Food Code. The average contamination levels of total viable bacteria were 3.11 (1.40-4.49) log CFU/g, and coliforms were detected in 5 out of 15 samples (1.18-1.85 log CFU/g). E. coli and S. aureus were not detected in all samples. Furthermore, the presence of 8 pathogens were not detected in all samples. The average contamination levels of lead, cadmium, and total mercury were 0.155 (0.079-0.264), 0.160 (0.040-0.287), and 0.017 (0.008-0.026) mg/kg, respectively. Benzo(a)pyrene, shellfish poison, and radioactivity were not detected in all samples. The results of this study suggest that the safety against all microbiological and chemical hazard factors in raw short-neck clams distributed in markets has been assured.

• Journal of Marine Life Science
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• v.1 no.1
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• pp.18-24
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• 2016

Since April 2012, doctor fish in the breeding tank and in the quarantine tank in Hanwha Aquaplanet Yeosu Aquarium have been dying, accompanied by diffuse bleeding around the mouth, in the chin, and at the bottom of the abdomen. In this study, the cause of death would be examined through the bacteriological study of doctor fish and the rearing water quality in the aquarium. The water quality and the bacterial counts of the rearing water in the exhibit tank and in the quarantine tank were analyzed once a week, starting from August to November 2014. Water quality was measured based on the following data: temperature was in the range of 24.5~26.8℃, pH at 6.77~7.94, DO at 6.15~8.61 ppm, ammonia at 0~0.93 ppm, nitrite at 0.009~0.075 ppm, and nitrate at 1.1~40.9 ppm. Studies revealed that the differences in these water quality factors were not related to the death of doctor fish. Bacterial counts in the rearing waters of Garra rufa slightly increased to 10³~10⁴ CFU/ml, just before the death of the doctor fish. Twelve strains of bacteria were isolated from the dead fish and rearing waters. The isolates were identified as Aeromonas veronii, Citrobacter freundii, Pseudorhodoferax aquiterrae, Shewanella putrefaciens, and Vibrio anguillarum on the basis of 16S rRNA gene sequences. The most dominant species was C. freundii, which showed medium sensitivity to florfenicol and norfloxacin, and was resistant to amoxacillin, doxycycline, oxytetracycline, tetracycline, and trimethoprim. Ten isolates were confirmed to be pathogenic to the doctor fish. Doctor fish infected with C. freundii and S. putrefaciens showed high mortality in the experimental groups. These results indicate that the variation in bacterial numbers in the rearing water was related to the death of doctor fish. C. freundii and S. putrefaciens were directly implicated in causing the death of doctor fish in the aquarium.

• Journal of Food Hygiene and Safety
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• v.21 no.1
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• pp.41-46
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• 2006

This study was carried out to survey the prevalence of foodborne pathogens in Sangshik products and their raw materials far the purpose of ensuring safety of those products in market, and establishing microbial regulatory standard. From 2002 to 2004, a total of 191 Sangshik products were purchased from market or mail-order sales, and major foodborne pathogens; E. coli, Salmonella, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Listeria monocytogenes, Campylobacter jejuni, Yersinia enterocolitica, E. coli O157:H7, Vibrio parahaemolyticus were tested. B. cereus, C. perfringens and E. coli were detected from 29 samples (15.2%), 21 samples (11.0%) and 1 sample (0.5%), respectively. But other tested bacteria were not detected. For the identification of contamination source, 53 Sangshik ingredients were collected from 9 different manufacture factories. The results were similar with the Sangshik products. Aerobic plate counts were ranging from $1.0X10^3cfu/g\;to\;1.5X10^8cfu/g$. B. cereus was detected from 13 samples (24.5%), and counted as less than 100 cfu/g. C. perfringens were detected from 2 samples (3.8%), and counted as less than 100 cfu/g. Other foodborne pathogens were not detected except for B. cereus and C. perfringens. From the results, it was revealed that potential of microbial hazard by Sangshik was relatively low. However, it would be suggested that hygienic management and controling be needed for the prevention of growing contaminated pathogens and cross contamination during process and sale due to improper storage and management.

• Journal of Life Science
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• v.22 no.5
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• pp.642-649
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• 2012

The southern coast of Korea is important for the ark shell ($Scapharca$ $broughtonii$) aquaculture, but the productivity was rapidly reduced during the previous decade by mass mortality. To overcome this economic loss, investigations only focused on environmental factors, and microbiological researches were performed insufficiently. In this study, two sites (Gangjin and Jinhae bay) were selected for their high and low rate of mortality, respectively, and the existence of microflora from underwater sediments in the bodies of $S.$ $broughtonii$ was analyzed. We screened the whole body of each sample and chose unique colonies, which exhibit alpha- and beta-hemolytic activity, for identification. The microflora in $S.$ $broughtonii$ was less variable than sediments, and restricted species were isolated. We identified 17 genera of 88 species and 16 genera of 64 species from the two bays, respectively. A major proportion was comprised of $Bacillus$ species, with the $Bacillus$ $cereus$ group being the most common species among the $Bacillus$ strains, while $Paenibacillus$, $Lynsilbacillus$, and $Vibrio$ species were the second most abundant species. At the genus level, there were no significant microbial differences between the two coastal regions. 64 species were isolated from rare site (Jinhae bay), but more species (88) with greater variety were isolated from the frequent site (Gangjin bay). Therefore, it was assumed that the cause of mass mortality lay in the difference in specie-level diversity, and conducting investigations on the diagnosis of pathogenic species by challenging tests using isolated unique species.

• Journal of Food Hygiene and Safety
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• v.23 no.3
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• pp.198-205
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• 2008

In this study, microbial investigation is accomplished for 554 Jeot-kal samples(102 of Jeot-kal, 448 of Seasoned Jeot-kal and 4 of Sik-khe, respectively) that corresponds with Coliform-bacteria, Escherichia coli, Aerobic live bacteria as hygienic indicator microorganisms, and Staphylococcus aureus, Vibrio parahaemolyticus as Food-borne pathogenic microorganisms, Based on the methods in Korea Food Code, reliable data are obtained as follows; in 31.9% rate of the samples, Coliform bacteria are verified in the extent of $0{\sim}20,000$ CFU/g as 2.3 logCFU/g. Especially, Seasoned Jeot-kal(37.7%,2.3 logCFU/g) are detected to 6 and 2 folds higher than those of Jeot-kal, 5.9% and 1.4 logCFU/g. Likewise, Escherichia coli is detected from 9 samples only in Seasoned Jeot-kal, that includes seasoned squid, seasoned octopus, seasoned roe of pollack, seasoned large-eyed herring and seasoned hairtail. Aerobic live bacteria are also detected in the range of $0{\sim}8.9{\times}10^8CFU/g$. Against salinity, E. coli are detected in samples only less than 10% salinity. Concomitantly, aerobic live bacteria count is decreased to $5.5{\sim}3.6$ log CFU/g upon the salinity is increased up to 25%. However, S. aureus and V. parahaemolyticus are not detected in 554 samples, presumptively referring Jeot-kal products are somehow free from such food-borne pathogens. As the results above, we deliberately consider that the sanitary control in Jeot-kal, which be necessarily fermented- as well as non-microbially inactivated should be ensured in near future and also suggest an effectual microbial standard corresponding to the Negativity in E. coli for Jeot-kal products.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.35 no.2
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• pp.115-121
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• 2002

To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.44 no.12
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• pp.1895-1904
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• 2015

Foodborne illness associated with food service establishments is an important food safety issue in Korea. In this study, foodborne pathogens (Bacillus cereus, Clostridium perfringens, Escherichia coli, pathogenic Escherichia coli, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Vibrio parahaemolyticus) and hygiene indicator organisms [total viable cell counts (TVC), coliforms] were analyzed for food and environmental samples from foodservice establishments at schools in Gyeonggi province. Virulence factors and antimicrobial resistance of detected foodborne pathogens were also characterized. A total of 179 samples, including food (n=66), utensil (n=68), and environmental samples (n=45), were collected from eight food service establishments at schools in Gyeonggi province. Average contamination levels of TVC for foods (including raw materials) and environmental samples were 4.7 and 4.0 log CFU/g, respectively. Average contamination levels of coliforms were 2.7 and 4.0 log CFU/g for foods and environmental swab samples, respectively. B. cereus contamination was detected in food samples with an average of 2.1 log CFU/g. E. coli was detected only in raw materials, and S. aureus was positive in raw materials as well as environmental swab samples. Other foodborne pathogens were not detected in all samples. The entire B. cereus isolates possessed at least one of the diarrheal toxin genes (hblACD, nheABC, entFM, and cytK enterotoxin gene). However, ces gene encoding emetic toxin was not detected in B. cereus isolates. S. aureus isolates (n=16) contained at least one or more of the tested enterotoxin genes, except for tst gene. For E. coli and S. aureus, 92.7% and 37.5% of the isolates were susceptible against 16 and 19 antimicrobials, respectively. The analyzed microbial hazards could provide useful information for quantitative microbial risk assessment and food safety management system to control foodborne illness outbreaks in food service establishments.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.1
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• pp.62-67
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• 1999

Infection rates of oyster ovarian parasite, Marteiliodes chungmuensis and productivity of the oyster shellstock infected with the parasite were investigated at the main seed collection areas in the southern coast of Korea where the extreme bad seed collection of oyster occurred in 1992 and 1993 to evaluate the cause of the bad seed collection. Additionally, the bacterial flora of the sea water and oyster lana were examined to identify the shellfish larva pathogenic bacteria like Vibrio sp. and Pseueomonas sp. In August 1992 to September 1993, infection rate of oyster ovarian parasite, M. chungmuensis at Tongyong, Kyongsangnam province, and Yosu, Chollanam province where the bad seed collection occurred, were $11.8\~100\%$ and $14.3\~100\%$, respectively. But the parasite was not detected in the shellstock collected at Daechon, Chungchongnam province. While a virus-like particle was identified in the cytoplasm of the egg infected by the parasite. The parasite infected egg was not able to fertilize completely. Uninfected egg in the gonad contaminated by the parasite could be able to fertilize but showed an abnormal development till D-shaped larva and then, died of necrosis after D-shaped lana. And some lana developed from low lipid content egg could not develop to the spat and died after the early umbo stage. The predominant bacteria in the oyster lana collected at bad seed collection areas were Pseudomonas sp. and Pseudomonas like bacteria and the occupancy rates were $53.3\~87.1\%$.

• Journal of Life Science
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• v.13 no.3
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• pp.333-342
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• 2003

The pathogenic effort of high glucose, possibly in concert with fatty acids, is mediated to vascular complications of diabetes via increased production of reactive oxygen species(ROS), reactive nitrogen species(RNS), and subsequent oxidative stress. This study was carried out to investigate the suppressive effect of buchu(Allium tuberosum) on oxidative stress in streptozotocin(STZ)-induced diabetes in Sprague Dawley male rats. The effect of buchu supplementation (10%) on lipid peroxidation, and antioxidative defense system in blood and liver was compared among normal rats fed basal diet(normal) and diabetic rats fed basal diet(DM-control) or 10% buchu-supplemented diet(DM-buchu). Diabetes was experimentally induced by the femoral muscle injection of 50 mg STZ per kg of body weight. Animals were sacrificed after 4 wks of experimental diets feeding. The induction of diabetes by STZ elevated the level of lipid peroxidation represented by thiobarbituric acid-reactive substances(TBARS) and conjugated dienes in plasma, LDL, liver, and erythrocytes. 10% buchu-supplemented diet significantly reduced the levels of conjugated dienes in erythrocytes(p<0.05) and lowered TBARS in liver and LDL to the levels of control. Induction of diabetes by STZ elevated Mn-superoxide dismutase(Mn-SOD) activity and lowered activities of glutathionine reductase(GSH-red) and glutathionine peroxidase(GSH-px). Catalase activity was not affected by the induction of diabetes by STZ. However, buchu supplementation to diabetic rats significantly elevated catalase activity(p<0.05) and slightly elevated GSH-px and GSH-red activities in liver. GSH levels of blood and liver were lowered or not changed by induction of diabetes by STZ, respectively, while buchu supplementation to diabetic rats significantly elevated hepatic GSH level (p<0.05). In conclusion, it can be concluded that buchu might be a food source to attenuate oxidative stress in diabetic patients by inhibiting lipid peroxidation, by increasing hepatic GSH level, and by inducing anti-oxidative enzyme systems.