• 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.

• Journal of fish pathology
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• v.21 no.1
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• pp.29-34
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• 2008

In this study, we examined in vitro antimicrobial activity of the hydrolysis compound (MBT-01108) of cactus stem (Opuntia ficus-indica var. saboten Makino) against 18 strains of fish pathogenic bacteria, Edwardsiella tarda, Listonella anguillarum, Vibrio sp., Lactococcus garvieae, L. raffinose, Streptococcus parauberis and S. iniae. Antibiotic susceptibility of the same bacteria was also assayed to commercial discs containing various antibacterials. According to these results, MBT-01108 showed the excellent antimicrobial activity against the tested pathogenic bacteria.

• Journal of fish pathology
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• v.14 no.1
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• pp.3-10
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• 2001

The present study was to obtain a basic research data about medicinal herbs by screening in vitro antimicrobial activity and the production of superoxide anion($O_2^-$) from the head kidney phagocytes of olive flounder, Paralichthys olivaceus. The following fourteen kinds of medicinal herbs extracted by boiling water were used : Gosam, Gwijeonu, Gujeolcho, Bagha, Bangpung, Yeongyo, Yagssug, Jiyu, Sambaegcho, Samjiguyeobcho, Sangbaegpi, Sohwehyang, Pyeonchug, Palgag. Antimicrobial activity against fish pathogenic bacteria, Listonella anguillarum, Vibrio sp., Vibrio alginolyticus, Edwardsiella tarda, Streptococcus sp. and Lactococcus garvieae, and the production of superoxide in kidney macrophage of olive flounder were examined by disk method and nitroblue tetrazolium(NBT) reaction, respectively. Among the tested herbs, Yagssug showed the highest antimicrobial activity against those fish pathogenic bacteria and stimulation of $O_2^-$ production.

• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.93-100
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• 2020

A bacterial strain inhibiting the growth of Vibrio anguillarum, the causative agent of vibriosis, was isolated from fish intestines. The isolated strain HS36 was identified as Aerococcus urinaeequi based on the characteristics of the genus according to Bergey's Manual of Systematic Bacteriology and by 16S rRNA sequencing. The growth rate and antibacterial activity of strain HS36 in shaking culture were higher than those in static culture, while the optimal pH and temperature for antibacterial activity were 7.0 and 30℃, respectively. The active antibacterial substance was purified from a culture broth of A. urinaeequi HS36 by Sephadex G-75 gel chromatography, Sephadex G-25 gel chromatography, and reverse-phase high-performance liquid chromatography. Its molecular weight, as estimated by Tricine SDS-polyacrylamide gel electrophoresis, was approximately 1,000 Da. The antibacterial substance produced by strain HS36 was stable after incubation for 1 h at 100℃. Although its antibacterial activity was optimal at pH 6-8, activity was retained at a pH range from 2 to 11. The purified antibacterial substance was inactivated by proteinase K, papain, and β-amylase treatment. The newly purified antibacterial substance, classified as a class II bacteriocin, inhibited the growth of Klebsiella pneumoniae, Salmonella enterica, and Vibrio alginolyticus.

• Journal of Aquaculture
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• v.14 no.4
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• pp.221-226
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• 2001

The ethanol extract of root bark of Morus alba strongly inhibited the Gram positive bacteria like Streptococcus sp., Lactococcus garvieae and Staphylococcus sp., but weakly the Gram negative bacteria like Listonella anguillarm and Edwardsiella tarda. It was more effective in liquid medium than in solid medium. The minimum inhibition concentration (MIC) of the extract in liquid medium was 19.8 and 790~1185 $\mu\textrm{g}$/$m\ell$ for the Gram positive and Gram negative bacteria, respectively. The extract concentration, at which the growth was totally inhibited, was 67.2~403.0 $\mu\textrm{g}$/$m\ell$ for the Gram positive bacteria but it was as high as 1185 $\mu\textrm{g}$/$m\ell$ for L. anguillarum and almost ineffective against E. tarda. For diet supplementation of the extract, effective soaking duration was 3 minutes. The fish diet soaked in the extract inhibited the growth of all the tested Gram positive strains, but not the Gram negative strains. The relationship between the weight of fish diet and absorption of the extract by the fish diet was Y=7.5757X + 4.6962($R^2$ = 0.9998).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.34 no.5
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• pp.545-549
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• 2001

To develop a natural antibacterial agent for fish bacterial diseases, antibacterial activity, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and bactericidal effect of cinnamon bark extract were examined against fish pathogenic bacteria. The antimicrobial effect of the extract to the fish diet was also estimated, Cinnamon bark extract showed the broad spectrum of antibacterial activity against fish pathogenic bacteria, especially, it had strong activity against Streptococcus iniae, Edwardsiella tarda and Listonella anguillarum. Its MIC was $75.8\sim189.6{\mu}g/mL$ against Cram positive bacteria, and $75.8\sim113.8{\mu}g/mL$ against Gram negative bacteria in liquid medium, It was found to show stronger bactericidal action against Gram negative bacteria than Cram positive bacteria. According to increasing concentrations of the extract, it resulted in a proportional reduction of viable cell counts of both S. iniae and L. anguillarum. The former was not detected by addition of $189.6{\mu}g/mL$ after 12 hours incubation and the latter by addition of $151.6{\mu}g/mL$ after 24 hours incubation, respectively. It was reasonable that fish diet was soaked in cinnamon bark extract for ten minutes. The relationship formula between the weight of fish diet and the extract absorbed to fish diet was Y=7.2726X+4.5083 ($R^2=0.9998$). The fish diet soaked in the extract inhibited the growth of all strains used in this study. Its antibacterial activity was stable at the range from $10^{\circ}C\;to\;35^{\circ}C$ during the storage period of 28 days. When the diet soaked in the extract was incubated in liquid medium at $35^{\circ}C$, it inhibited the growth of microorganisms inhabited in the diet.

• 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.