• Title/Summary/Keyword: Vibrio harveyi

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Functional characterization and expression analysis of c-type and g-like-type lysozymes in yellowtail clownfish (Amphiprion clarkii)

  • Gaeun Kim;Hanchang Sohn;WKM Omeka;Chaehyeon Lim;Don Anushka Sandaruwan Elvitigala;Jehee Lee
    • Fisheries and Aquatic Sciences
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    • v.26 no.3
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    • pp.188-203
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    • 2023
  • Lysozymes are well-known antibacterial enzymes that mainly target the peptidoglycan layer of the bacterial cell wall. Animal lysozymes are mainly categorized as g-type, c-type, and i-type based on protein sequence and structural differences. In this study, c-type (AcLysC) and g-like-type (AcLysG-like) lysozymes from Amphiprion clarkii were characterized in silico via expressional and functional approaches. According to in silico analysis, open reading frames of AcLysC and AcLysG-like were 429 bp and 570 bp, respectively, encoding the corresponding polypeptide chains with 142 and 189 amino acids. Elevated expression levels of AcLysC and AcLysG-like were observed in the liver and the heart tissues, respectively, as evidenced by quantitative real-time polymerase chain reaction assays. AcLysC and AcLysG-like transcript levels were upregulated in gills, head kidney, and blood cells following experimental immune stimulation. Recombinant AcLysC exhibited potent lytic activity against Vibrio anguillarum, whereas recombinant AcLysG-like showed remarkable antibacterial activity against Vibrio harveyi and Streptococcus parauberis, which was further evidenced by scanning electron microscopic imaging of destructed bacterial cell walls. The findings of this study collectively suggest the potential roles of AcLysC and AcLysG-like in host immune defense.

Construction and Characterization of a Recombinant Bioluminescence Streptomycetes for Potential Environmental Monitoring

  • Park, Hyun-Joo;Hwang, Keum-Ok;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.706-709
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    • 2002
  • Bacterial bioluminescence has been known to be a highly valuable reporter system for its potential application as an effective and simple environmental monitoring method for toxic compounds. In this short report, we constructed a streptomycetes-Escherichia coli shuttle vector-containing bioluminescence system and evaluated its potential application for toxic compounds monitoring. The luxAB biolurninescence genes from Vibrio harveyi were cloned into a streptornycetes-E. coli shuttle vector (named pESK004) and functionally expressed in Streptomyces lividans. The recombinant S. lividans containing pESK004 exhibited an optimal biolurninescence at the optical density ($OD_{600\;nm}$) of 0.4-0.5 and aldehyde concentration of 0.005%. When the recombinant bioluminescence streptomycetes was exposed to a toxic compound such as heavy metals, chlorinated phenols, or pesticides, the bioluminescence was decreased proportionally to the concentration of toxic compound in the assay mixture. The $EC_{50}$ (effective concentration to decrease 50% of the bioluminescence prior to exposure) values in the recombinant biolurninescence streptomycetes for mercury, 2,4-dichlorophenol, and malathion were measured at 2.2 ppm, 144.0 ppm, and 82.4 ppm, respectively. The degree of sensitivity and specificity pattern toward these toxic compounds characterized in this recombinant bioluminescence streptomycetes were unique when compared with previously reported bacterial bioluminescence systems, and this revealed that a recombinant bioluminescence streptomycetes might provide an alternative or complementary system for potential environmental monitoring.

Supplemental effects of biofloc powder on growth performance, innate immunity, and disease resistance of Pacific white shrimp Litopenaeus vannamei

  • Lee, Chorong;Kim, Soohwan;Lim, Se-Jin;Lee, Kyeong-Jun
    • Fisheries and Aquatic Sciences
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    • v.20 no.7
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    • pp.15.1-15.7
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    • 2017
  • An 8-week feeding trial was conducted to study the effect of dietary supplementation of a biofloc powder on growth performance and non-specific immune response of Litopenaeus vannamei. Seven experimental diets were prepared with supplementation of graded levels of dried biofloc powder by 0, 0.5, 1.0, 2.0, 4.0, 6.0, and 8.0% (designated as Con, BF0.5, BF1, BF2, BF4, BF6, and BF8, respectively). Triplicate groups of shrimp ($1.01{\pm}0.01g$) were hand-fed with one of the diets four times a day. At the end of the feeding trial, significantly (P<0.05) higher growth performance and feed utilization were obtained in BF4 groups compared to those fed the Con diet. The innate immunity of shrimp was improved by the dietary supplementation of biofloc. Dietary inclusion of biofloc at the level of 4.0% significantly increased disease resistance of shrimp against Vibrio harveyi. The results indicate that biofloc might be used as a dietary supplement for growth performance, innate immunity and disease resistance of Pacific white shrimp.

Antibacterial activity and toxicity of Halymenia durvillei red seaweed from Kayangan island, South Sulawesi, Indonesia

  • Kasmiati, Kasmiati;Nurunnisa, Andi Tenri;Amran, Amran;Resya, Muhammad Ikhwan;Rahmi, Mufti Hatur
    • Fisheries and Aquatic Sciences
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    • v.25 no.8
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    • pp.417-428
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    • 2022
  • This study aimed to determine the antibacterial activity and toxicity of methanol and hexane extracts of Halymenia durvillei red seaweed which were found abundantly in Kayangan island, South Sulawesi. The antibacterial activity of the crude extract was tested against five gram-negative bacteria, namely Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Aeromonas hydrophila, and Vibrio harveyi at a dose of 200 g/disk. Extract toxicity was tested on Artemia salina larvae at concentrations of 1,000, 500, 250, 125, 62.5, and 31.25 ㎍/mL. The results showed that the methanol and hexane extracts of H. durvillei had the highest activity against S. thypi and A. hydrophila, respectively, with inhibition zones of 26.2 mm and 21.0 mm. On the other hand, the two extracts did not show activity against E. coli and P. aeruginosa, respectively. The toxicity of the methanol extract of H. durvillei was twice as high as that of the hexane extract with half-maximal inhibitory concentration of 98.24 and 184.21 ㎍/mL, respectively. Thus, the methanol and hexane extracts of red seaweed H. durvillei have the potential as new antibacterial agents respectively against the pathogenic bacteria S. typhi and A. hydrophila, but also have the opportunity to be developed into antitumor herbal compounds.

Bacterial Community Analysis and Antibacterial Activity Isolated from Umbraulva japonica (초록갈파래(Umbraulva japonica)에서 분리한 세균의 군집 구조 분석 및 항균 활성)

  • Kim, Ji-Hyun;Park, So-Hyun;Moon, Kyung-Mi;Kim, Dong-Hwi;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.127-134
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    • 2018
  • In this study, 79 bacterial isolates were collected from the surface of marine algae Umbraulva japonica. As a result of analysis of 16s rRNA gene sequence, the 79 isolated bacteria were divided into 4 major groups: [Proteobacteria (74.69%), Actinobacteria (2.53%), Fimicutes (2.53%), and Bacteroidetes (20.25%)] - 7 classes (Actinobacteria, Flavobacteria, Sphingobacteria, Baciili, Alphaproteobacteria, Betaproteobacteria, and Gammaproteobacteria), 12 orders, 17 families and 31 genera. The newly isolated 3 strains could be novel species because of less than 97% similarity in 16s rRNA sequence. Therefore, it is considered that additional experiments should be conducted together with the standard strain. Analysis of 79 bacterial antibacterial activity against human and fish pathogens, such as Edwardsiella tarda, Vibrio harveyi, Streptococcus iniae, Steptococcus parauberis, Escherichia coli, Steptococcus mutans, Listeria monocytogenes and Vibrio vulnificus, was performed by using the supernatant liquid and pellet. As a result, pellet of UJT9, UJT20 and UJR17 showed antibacterial activity against V. vulnificus, UJR17 also showed antibacterial activity against S. parauberis. UJT7 and UJT20, UJR17 have been identified as Bacillus sp. and Pseudomonas sp. and it may be safely assented that it's beneficial to carry out additional experiments for various applications.

Industrial Application for Aquaculture of Fermented Soybean with Bacillus polyfermenticus SCD (양식업에서 바실러스 폴리퍼멘티쿠스 콩발효물의 상업적 응용)

  • Lee, Jin Young;Kim, Kang Min;Kang, Jae Seon
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.1
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    • pp.48-54
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    • 2007
  • Bacillus polyfermenticus SCD (B. polyfermenticus SCD) has been appropriately used for the therapy of long-term intestinal disorders, because live strains in the form of active endospores can successfully reach the target intestine in humans. B. polyfermenticus SCD produces the most antibiotic-related materials. In the present study, B. polyfermenticus SCD was fermented with soybeans (BPFS) and its probiotic properties were investigated. B. polyfermenticus SCD and BPFS showed a broad spectrum of antimicrobial activity against pathogenic Gram-positive (Streptococcus parauberis, Streptococcus iniae, Lactococcus garviae) and Gram-negative (Flexibacter tractuosus, Vibrio harveyi, Vibrio vulnificus, Vibrio ordalii) bacteria and moulds (Aspergillus niger, Aspergillus oryzae). Sebastes schlegeli were used to examine survival rate and cleanup action by BPFS. Bacterial infection resulted in a mortality of up to 99% in the commercial fodder fed groups. BPFS both enhanced the growth rate of fry by improving their appetite and had cleaned by decreased eutrophication. Therefore, it seems appropriate that BPFS should be developed as an antibiotic replacement, favorable fodder additive, and antifungal material in fish farming systems.

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Degradation capability of macromolecular organic matters and antimicrobial activities of Bacillus species isolated from surf clam (Tresus keenae) (왕우럭(Tresus keenae)에서 분리된 Bacillus species의 고분자 유기물질 분해능력과 항균활성)

  • Yi, Seung-Won;Moon, Sung-Hyun;Cho, Ho-Seong;Kim, Chul-Won
    • Korean Journal of Veterinary Service
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    • v.40 no.4
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    • pp.265-275
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    • 2017
  • The production of enzymes that help digestion, assimilation of essential nutrients, and prevent pathogenic bacteria are important for probiotics used in aquaculture. The objective of this study was to investigate enzyme activities for macromolecular organic matters and antimicrobial properties of the selected potential probiotics isolated from gut of surf clam (Tresus keenae) against well-known shellfish-pathogenic bacteria. Among 65 isolates from guts of 60 surf clams, seven Bacillus strains with outstanding degradation capability of macromolecule organic matter were selected as potential probiotics as follows: TKI01 (B. vietnamensis), TKI02, TKI26 (B. thuringiensis), TKI14, TKI32, TKI42 (B. amyloliquefaciens), and TKI18 (B. stratosphericus). After in vitro antimicrobial activity test was performed against five shellfish-pathogenic bacteria including Listonella anguillarum, Vibrio parahaemolyticus, V. splendidus, V. harveyi, V. tubiashii, PCR assay was performed to detect bacteriocin-producing strain. PCR results revealed that the five Bacillus strains possessed diverse bacteriocin genes including ericinA, coagulin, surfactin, iturin, bacyllomicin, fengycin, bacylisin, subtilin, and lantibiotics. In the present study, the selected seven Bacillus strains showed different enzyme activities according to types of macromolecule organic matters. And their antimicrobial activities varied based on the species of pathogenic bacteria. In addition, at least five Bacillus strains had genetic potential to produce several natural lipopeptide antibiotics that may help biological control of surf clam aquaculture. Therefore, mixed use of probiotics might show co-operative effect and increase the efficiency of probiotics rather than separate use. To the best of our knowledge, it is the first report on antimicrobial properties of Bacillus species isolated from surf clam.

Effect of Fermented Soybean Meal by Bacillus subtilis in Pacific White Shrimp Litopenaeus vannamei Diets on Growth Performance, Innate Immunity and Disease Resistance (Bacillus subtilis 발효대두박의 사료 내 첨가가 흰다리새우(Litopenaeus vannamei)의 성장, 면역력 및 질병저항성에 미치는 영향)

  • Kim, Soohwan;Lee, Chorong;Chang, Kyunghoon;Bae, Junyoung;Cho, Seong-Jun;Lim, Se-Jin;Lee, Kyeong-Jun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.1
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    • pp.8-14
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    • 2017
  • A feeding trial was conducted to evaluate the effect of dietary supplementation with fermented soybean meal by Bacillus subtilis (BFSBM) on growth performance, feed utilization, innate immunity, water quality, and disease resistance of Litopenaeus vannamei. A basal fish meal-based diet was regarded as a control and two other diets were prepared by replacing 30% fish meal with either soybean meal (SBM) or BFSBM. Triplicate groups of shrimp ($0.80{\pm}0.010g$) were fed one of the experimental diets for 6 weeks. No differences were found in final body weight, feed conversion ratio, or survival among all dietary treatments after the feeding trial. Shrimp fed the BFSBM diet had significantly higher phagocytic and phenoloxidase activities than those of shrimp fed the SBM diet. Total ammonia concentrations in culture water during a zero water exchange test tended to decrease in tanks where BFSBM diet was provided. Disease resistance of shrimp to Vibrio harveyi tended to increase in the BFSBM group during 14 days of challenge compared to that in the control (CON) and SBM groups. These results show that dietary supplementation with BFSBM can enhance the innate immunity of Pacific white shrimp and replace at least 30% of dietary fish meal.

Effect of Mixed Oxidants and Sodium Hypochlorite on Pathogenic Microorganisms in Olive flounder Paralichthys olivaceus Aquaculture on Jeju Island (제주도 양식 넙치(Paralichthys olivaceus)에서 분리한 병원균 3종에 대한 Mixed Oxidant 및 차아염소산나트륨 살균효과)

  • Park, Cheonman;Kim, Ki-hyuk;Moon, Hye-na;Yeo, In-Kyu
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.4
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    • pp.389-396
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    • 2018
  • Marine pathogenic bacteria, such as Streptococcus parauberis, Edwardsiella tarda and Vibrio harveyi, can cause lethal infections in farmed fish, ozone and antibiotics, are employed to sterilize waters used for rearing fish to mitigate this threat. The most widely used method is treatment with sodium hypochlorite solution. However, the maintenance of a constant concentration of chlorine in rearing waters can be difficult. We investigated the potential of a mixed oxidant (MO) solution generated by electrolysis of sea water to improve water quality. We measured the survival rates of fish pathogenic bacteria exposed to different concentrations of MO (0.5, 1.0, 1.5 and 2.0 MO) and sodium hypochlorite (0.5, 1.0, 1.5 and 2.0 ppm) for various lengths of time (0, 5, 10, 15, 20, 25 and 30 min). We found a time-dependent decrease in the survival rates of the tested pathogenic microorganisms. The sterilization effect of the MO solution on pathogenic organisms was greater than that of sodium hypochlorite for gram-negative and gram-positive bacteria. We conclude that MO solution produced by electrolysis could be used to maintain a constant chlorine concentration in aquaculture systems.

Introduction of bacterial and viral pathogens from imported ornamental finfish in South Korea

  • Choi, Hee Jae;Hur, Jun Wook;Cho, Jae Bum;Park, Kwan Ha;Jung, Hye Jin;Kang, Yue Jai
    • Fisheries and Aquatic Sciences
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    • v.22 no.2
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    • pp.5.1-5.9
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    • 2019
  • Background: Live fish import may lead to the unintended introduction of pathogens. We examined the monthly distribution of microbial pathogens in ornamental finfish imported into South Korea over a 6-month period. Results: Vibrio alginolyticus was detected in one lemon damsel in June and July; V. vulnificus was detected in one lemon damsel, one caerulean damsel, and one pearl-spot chromis and one ocellaris clownfish in July, April, and May, respectively; Photobacterium damselae was detected in one ocellaris clownfish and one caerulean damsel in June and July, respectively; V. anguillarum was detected in one pearl-spot chromis in February; V. harveyi was detected in one ocellaris clownfish and two mandarin fish in February and April, respectively; Yersinia ruckeri was detected in a pearlscale goldfish group in June and July and in two colored carp groups in July; and Lactococcus garvieae was detected in a lemon damsel group and a sutchi catfish group in July and May, respectively. European catfish virus, the only viral pathogen detected, was found in two sutchi catfish groups in May. Conclusion: This study is the first to identify pathogenic species and the presence or absence of pathogens (non-quarantine diseases) in imported ornamental finfish. These results demonstrate that various pathogens with the potential to harm indigenous fish populations can accompany ornamental finfish imported into South Korea.