• Fisheries and Aquatic Sciences
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• v.24 no.10
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• pp.319-329
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• 2021

In the search for antibiotic alternatives from safe and effective medicinal plants against fish pathogenic bacteria, we found that a combined extract (CE) of 1:1 (w/w) ratio of Angelica gigas Nakai roots and aerial parts of Artemisia iwayomogi Kitamura showed antibacterial activity against the fish pathogenic bacteria. By antibacterial activity-guided fractionations and isolations, five compounds were isolated and identified as decursinol angelate (1), decursin (2), xanthotoxin (3), demethylsuberosin (4), and 2,4-dihydroxy-6-methoxyacetophenone (5) through spectroscopic analyses, such as nuclear magnetic resonance (NMR) and mass spectrometry (MS). Among the compounds, 1 and 2 showed the highest antibacterial activities against Streptococcus iniae and Vibrio anguillarum, showing minimum inhibitory concentrations (MICs) of 62.5-250 ㎍/mL. Compounds 3, 4, and 5 were also found to be active, with MICs of 31.25-1,000 ㎍/mL for those strains. Furthermore, active compounds, 1 and 2 in CE were simultaneously quantified using high-performance liquid chromatography-tandem MS (HPLC-MS/MS). The average contents of 1 and 2 in CE was 3.68% and 6.14%, respectively. The established method showed reliable linearity (r² > 0.99), good precision, accuracy, and specificity with intra- and inter-day variations of < 2 % and recoveries of 90.13%-108.57%. These results may be helpful for establishing the chemical profile of CE for its commercialization as an antibiotic alternative in aquaculture.

• Journal of fish pathology
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• v.37 no.1
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• pp.133-138
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• 2024

This study is a report on combinational effects between four sesquiterpene lactones (SLs) from bay laurel (Laurus nobilis) leaves, and oxytetracycline (OTC) or amoxicillin (AMX) against four fish pathogenic bacteria such as Vibrio anguillarum, V. harveyi, Edwardsiella tarda, and Streptococcus iniae. Individually, four SLs exerted little antibacterial activity against fish pathogenic bacteria. However, when combined with OTC or AMX, they showed synergistic interaction against pathogenic bacteria. Especailly, zaluzanin C (1) reduced the MIC of OTC (or AMX) eight-fold. Our results showed that combinations of SLs with antibiotics (ABTs) are more effective than ABTs alone to control pathogenic bacteria. The highest synergistic effect was observed when zaluzanin C (1) was combined with OTC or AMX against V. harvey or S. iniae, displaying significant reductions of MICs up to 8-fold (0.125 to 0.015 ㎍/mL and 0.0078 to 0.0009 ㎍/mL). In addition, zaluzanin C (1) improved the antibiotic potency of OTC against OTC resistant V. harveyi (250 ㎍/mL to 62.5 ㎍/mL). Synergism between ABTs and phytochemical such as SLs could be a therapeutically helpful concept to improve the efficacy of ABTs and prevent antibiotic resistance. These results suggest that SLs can be used as an alternative to reduce antibiotic resistance in aquaculture.

• Journal of fish pathology
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• v.22 no.3
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• pp.317-326
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• 2009

This study has shown the screening of anti-bacterial activity of three Indian medicinal plant choloroform : methanol (50:50) solvent leaf extracts (i.e. Azadirachta indica, Ocimum sanctum, and Curcuma longa) with different concentrations (10, 5, 2.5, 1.25, 0.625, 0.312, and 0.156 mg/ml) under in vitro conditions against fish pathogenic bacteria, Aeromonas hydrophila, Streptococcus iniae, Vibrio harveyi, V. anguillarum, and Edwardsiella tarda isolated from olive flounder farms, Jeju Island, South Korea. The anti-microbial activity of the A. indica and O. sanctum extracts yielded the zones of growth inhibition (ZI) was 3 and 1mm against A. hydrophila at concentration of 0.156 mg/ml when compared to that of tetracycline standard (3 mm). At highest concentration (10 mg/ml) of A. indica, O. sanctum, and C. longa, high inhibition was 9, 7, and 6 mm when compared to that of tetracycline (11 mm) against A. hydrophila. The minimum inhibitory concentration (MIC) of A. indica, O. sanctum, and C. longa at 0.156 mg/ml that yield 9, 10, and 13 CFU/ml for A. hydrophila, 16, 22, and 25 CFU/ml for S. iniae and 18, 22, and 23 CFU/ml for E. tarda compared to the tetracycline. At highest concentration (10 mg/ml) of the three extracts was better inhibiting the growth of A. hydrophila, S. iniae and E. tarda. A. indica, O. sanctum, and C. longa were determined to the potential antioxidant activityon the basis of their scavenging activity of the stable 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical. A. indica extract was 0.625 mg/ml which indicated that the strong anti-oxidant activity. However, O. sanctum and C. longa extracts showed weak anti-oxidant activity at this concentration. Hence, in vitro assay among the pathogens, A. hydropila is better inhibitory activity of the extracts. It is evident that the Indian medicinal plants extracts were subjected to its effectiveness against A. hydrophila, S. iniae, and E.tarda at low concentrations. The obtained results in the present study suggested that the Indian plant extracts is a prevention tools for Korean olive flounder aquaculture pathogens and its need further advance investigation.

• 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.