• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.130-138
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• 2016

The use of antibiotics in aquaculture raises environmental and food safety concerns because chronic exposure of an aquatic ecosystem to antibiotics can result in the spread of antibiotic resistance, bioaccumulation of antibiotics in the organisms, and transfer of antibiotics to humans. In an attempt to overcome these problems, high-concentration red clay was applied as an alternative antibiotic against the following common fish pathogens: Aeromonas salmonicida, Vibrio alginolyticus, and Streptococcus equinus. The growth of A. salmonicida and V. alginolyticus was retarded by red clay, whereas that of S. equinus was promoted. Phase contrast and scanning electron microscopy analyses confirmed the attachment of red clay on cell surfaces, resulting in rapid gravitational removal and cell surface damage in both A. salmonicida and V. alginolyticus, but not in S. equinus. Different cell wall properties of grampositive species may explain the unharmed cell surface of S. equinus. Significant levels of oxidative stress were generated in only the former two species, whereas significant changes in membrane permeability were found only in S. equinus, probably because of its physiological adaptation. The bacterial communities in water samples from Oncorhynchus mykiss aquacultures supplemented with red clay showed similar structure and diversity as those from oxytetracycline-treated water. Taken together, the antibiotic effects of high concentrations of red clay in aquaculture can be attributed to gravitational removal, cell surface damage, and oxidative stress production, and suggest that red clay may be used as an alternative for antibiotics in aquaculture.

• 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.

• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.513-518
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• 1999

A total of 113 Vibrio sp. strains were examined for plasmid content which were subjected to digestion with restriction enzymes. About the 55% Vibrio spp. have the plasmid more than one. Most of these plasmid various derivatives ranged from $2.4\;kb{\sim}23\;kb$, especially two strains of V. mimicus and one strain of V. furnissii carried one high-molecular weight plasmid (molecular weight ranging between $70\;kb{\sim}100\;kb$). Results of restriction analysis for plasmid of this three strains were by no means the rule. For detection of tdh and ctx gene, the virulence factor involved in the pathogenesis, we carried out the TDH and CT assay, PCR amplification, and hybridization. A total 11 strains were produced TDH, involved in 9 strains of V. parahaemolyticus and 1 strain of V. alginolyticus from clinical isolates and 1 strains of V. mimicus from environmental isolates. In the experiments of tdh gene detection, in all, 3 strains of V. parahaemolyticus from clinical isolates and 2 strains from environmental isolates could be successfully amplified in 400 bp by PCR. The PCR results were consistent with DNA hybridization tests. In the experiments of CT assay, in all, 3 strains of V. cholerae from clinical isolate and 1 strain of V. cholerae from environmental isolates were observed CT-producing. These CT-producing strains amplified in 302 bp by PCR for the detection of ctx gene. All CT-producing strains hybridized with digoxigenin-labeled DNA probe, while CT-negative strains did not hybridize. Also hybridization tests results for detection of ctx gene consistent with PCR.

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1080-1086
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• 2005

Membranes prepared from Vibrio natriegens oxidized both NADH and deamino-NADH as substrates. The maximum activity of the membrane-bound NADH oxidase was obtained at about pH 8.5 in the presence of 0.2 M NaCl, whereas that of the NADH:ubiquinone oxidoreductase was obtained at about pH 7.5 in the presence of 0.2 M NaCl. Electron transfer from NADH or deamino-NADH to ubiquinone-l or oxygen generated a considerable membrane potential (${\Delta}{\psi}$), which occurred even in the presence of $20{\mu}M$ carbonylcyanide m-chlorophenylhydrazone (CCCP). However, the ${\Delta}{\psi}$ was completely collapsed by the combined addition of $10{\mu}M$ CCCP and $20{\mu}M$ monensin. On the other hand, the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were inhibited by about $90\%$ with $10{\mu}M$ HQNO, whereas the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were inhibited by about $60\%$. Interestingly, the activity of the NADH:ubiquinone oxidoreductase and the ${\Delta}{\psi}$ generated at the NADH:ubiquinone oxidoreductase segment were resistant to the respiratory chain inhibitors such as rotenone, capsaicin, and $AgNO_3$, and the activity of the NADH oxidase and the ${\Delta}{\psi}$ generated by the NADH oxidase system were very sensitive only to $AgNO_3$. It was concluded, therefore, that V. natriegens cells possess a $AgNO_3$-resistant respiratory $Na^+$ pump that is different from the $AgNO_3$-sensitive respiratory $Na^+$ pump of a marine bacterium, Vibrio alginolyticus.

• Fisheries and Aquatic Sciences
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• v.5 no.3
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• pp.178-182
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• 2002

The authors identified 68 Vibrio strains from Gwangan beach seawater from June to October in 2001. We identified them as 19 strains of Vibrio alginolyticus, 15 strains of V. vulnificus, 15 strains of V. parahaemolyticus, 11 strains of V. cholerae non O1, 7 strains of V. fluvialis and just one strain of V. hollisae. They showed their typical biochemical characteristics by API 20E kit (bioMerieux), respectively. It was examined whether their cultural supernatants had enzymatic activities such as hemolysin, protease or urease. The 46 strains showed hemolytic activities and/or protease activities. But we could not find any strain which had urease activity. All isolates of V. cholerae non O1 showed $\beta$ hemolysis. The others showed $\alpha$ hemolysis or did not show clear zones on sheep blood agar plates. These results of Kanagawa phenomenon were not always correspondant with hemolytic activities of cultural supernatants at late log phase. Some strains had higher hemolytic activities despite of showing protease activities on skim milk agar plates and in litmus milk media. On the other hand, some strains showed protease activities but did not show hemolytic activities. Therefore we could guess that there were the relationships between hemolysins and proteases produced by pathogenic vibrios.

• Fisheries and Aquatic Sciences
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• v.7 no.1
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• pp.10-15
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• 2004

A total of 52 pathogenic Vibrio strains was isolated from the Gwangan Beach during summer in 2003. The isolated vibrios were composed of 6 different species: V. parahaemolyticus, V. cholerae non O1, V. fluvialis, V. vulnificus, V. alginolyticus, and V. mimicus. V. parahaemolyticus was most predominant as $46\%$ (24/52), V. cholerae non O1 was the second with $23\%$ (12/52), and V. fluvialis was the third with $17\%$ (9/52). Among the isolated strains, 22 strains showed hemolytic, proteolytic or ureolytic activity. Eight strains showed both hemolysin and protease activities, and either 6 strains showed only hemolysin activities and 7 strains only protease activities. Only one strain of V. parahaemolyticus isolates showed urease activity. The urease-positive V. parahaemolyticus strain (V. parahaemolyticus S25) showed the same biochemical characteristics as the reference strain, V. parahaemolyticus KCTC 2471 (urease­negative) except for urease production. To compare the degree of virulence of Vibrio strains having different pathogenic factors, hemolysin, protease, or urease-positive strains were injected into groups of 10 each of ICR mice (7- to l0-week-old male). The lethal rate of urease-positive V. parahaemolyticus S25 was significantly high, being $70\%$. Protease-positive strains showed $40-60\%$ of lethal rate. Hemolysin-positive strains showed no mortality, similar to non-pathogenic V. parahaemolyticus KCTC 2471 and V. parahaemolyticus FM12.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.1
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• pp.17-25
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• 1996

The results identified for bacterial flora 174, Vibrio spp.132, and coliform group 183 strains isolated from the areas of last China Sea and Yosu coastal sea during from August 6th. to 14th. 1992 were as follow: 40 strains among the 74 strains of bacteria flora isolated from fast China sea area were Pseudomonas spp.$(54\%)$ and 60 strains among the 100 strains isolated from Yosu sea area were Enterobacteriaceae $(60\%)$. Four strains were Vibrio alginoliticus and one strain of V, parahaemolyticus among 5 strains of genus Vibrio isolated from last China Sea. While 54 strains were V. alginolyticus $(43\%)$ and V, parahaemolyticus $(17\%)$ among 127 strains genus Vibrio isolated from Yosu coastal sea area. Seventy nine strains among the 156 strains of coliform group isolated from Vosu sea area were Escherichia coli I $(51\%)$ and each one strain Citrobacter freundii I and II. 3 strains among 27 strains isolated from last China sea area were E. coli$(11\%)$ and 1 strain of C. freundii I. Coliform group was grouped into 16 types by IMViC system, $44^{\circ}C$, gelatin liquefaction test.

• Journal of fish pathology
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• v.27 no.1
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• pp.17-24
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• 2014

To study the possible use of probiotics in fish farming, The in vitro and in vivo antibacterial effects of Pseudomonas aeruginosa MB I-3 (MB I-3) against the fish pathogenic bacterium Listonella anguillarum were evaluated. The inhibitory effects of MB I-3 against vibrios were investigated by the double layer method and the co-culture. The results showed that MB I-3 inhibited the growth of pathogenic vibrios including Listonella anguillarum, Vibrio alginolyticus, Vibrio cholerae, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio parahaemolyticus and Vibrio vulnificus. Extracellular substances obtained from the cultural supernatant of MB I-3 by ethyl acetate extraction showed inhibitory effects on L. anguillarum. The antibacterial substance of MB I-3 was evaluated to destroy the cell membrane of L. anguillarum in electron micrographs. The probiotic effects of MB I-3 was tested by exposing olive flounder (Paralichthys olivaceus) fry to L. anguillarum with or without MB I-3. The cumulative mortality of olive flounder fry infected with L. anguillarum was 24% in the group with MB I-3, while it was 46% in the control group without MB I-3. These results indicate that MB I-3 has potential applications as a probiotic for the control of fish pathogenic vibrios in fish rearing system.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2001.10a
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• pp.239-240
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• 2001

Collagenases are generally defined as enzymes capable of degrading the polypeptide backbone of native collagen under conditions which do not denature the protein. Two types of proteases with collagenolytic activity have been reported and thought to play different physiological functions. Metallo-collagenases, firstly discovered in tadpole tissue explants are zinc-containing enzymes requiring calcium for optimum activity and stability, and These enzymes have been widely studied from various mammalian tissues as well as from bacteria, such as Bacillus cereus, Clostridium histolyticum, Achromobacter, Vibrio alginolyticus and Clostridium perfringens and snake venoms. (omitted)

• Korean Journal of Ichthyology
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• v.29 no.1
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• pp.41-51
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• 2017

To understand the application in farm for the fish aquaculture, we investigated biological and pathological traits on red sea bream Pagrus major which were reared in each copper-alloy net cage and the synthetic fiber net cage for 9 months. Two groups of cage were made and set in Yokji-eup, Tongyoung, Gyeongsangnam-do in size of 25 m in diameter and 10 m of depth. Survival rate of the red sea bream in the rearing copper-alloy net cage and synthetic fiber cage showed 99.75% and 99.70% respectively, there was no significant difference. Daily weight growth rate in each net was shown to 2.13 g/day and 1.65 g/day. Health analysis by blood composition analysis showed a favorable result in the copper-alloy net cage rather than in the synthetic fiber net. Bioaccumulation of heavy metal such as Cu and Zn especially in gonad was higher than other organ. Bioaccumulation of Cu and Zn in the muscle was lower compared to the permitted standard for food safety. Pathogenic infection test discovered Microcotyle tai for parasite, V. alginolyticus and other five species for bacteria. But there was a little bit difference of bacteria infection in copper-alloy net cage and copper-alloy net cage is expected to be has antibacterial effect. Thus, copper-alloy net cage can be applied to farm considering its system stability, recycling, antibiosis and food safety.