• Microbiology and Biotechnology Letters
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• v.25 no.2
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• pp.151-158
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• 1997

For the production of potent chitinolytic enzyme from bacteria, screening was carried out. Of 100 samples from soil, fresh water and sea water collected from the Kyung-gi area, 7 strains of chitinolytic bacteria were isolated. Among them, Aeromonas hydrophila 5-3K showed the highest chitinolytic activity. Culture conditions of Aeromonas hydrophila for the production of chitinolytic enzyme were inverstigated and lytic enzyme was fractionated by the use of ammonium sulfate and Sephadex G-100. Maximum production of chitinolytic enzyme was obtained at pH 7.0 and 30$\circ$C with chitin concentration between 0.2% and 1.0%. Conditions for the enzyme production were optimized including fermentor cultivation. The chitinolytic system of Aeromonas hydrophila 5-3K was composed of two enzymes, chitinase and chitobiase.

• Applied Biological Chemistry
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• v.44 no.4
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• pp.251-256
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• 2001

To develop biofertilizer solubilizing inorganic phosphate, a bacterium having high abilities to solubilize inorganic phosphate were isolated from cultivated soils. The strain was identified to Aeromonas hydrophila DA57, based on the physiological and biochemical properties. The optimum temperature and initial pH to solubilize insoluvle phosphate in sucrose minimal medium were $30^{\circ}C$ and pH 7.0, respectively. In these conditions phosphate solubilizing activities of the strain against three types of insoluble phosphate were quantitatively determined. It was possivle to distinguish between solubilization through release of gluconic acid and still unknown mechanism. Aemmonas hydrophila DA57 harbored a 4.5 kb cryptic plasmid.

• Korean Journal Plant Pathology
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• v.11 no.3
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• pp.258-264
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• 1995

Production and some properties of chitinolytic enzymes were investigated by 80% ammonium sulfate precipitates (crude enzymes) from culture supernatant of antagonistic bacteria, Chromobacterium violaceum strain C-61 and strain C-72, Aeromonas hydrophila, Aeromonas caviae, and Serratia marcescens. The maximum production of chitinase was obtained from the 3-day culture at 28$^{\circ}C$ in C. violaceum stains, the 6-day culture in S. marcescens, and the 2-day culture in A. hydrophila and A. caviae. In the optimum culture periods, chitinase activity of C. violaceum strains C-61 was 1.5, 5.5, 12.0 and 11.3 times higher than those of strain C-72, S. marcescens, A. hydrophila and A. caviae, respectively. However, N,N'-diacetylchitobiase activity was 3.2 times higher in S. marcescens than in C. violaceum strain C-61, and that of Aeromonas spp.was very low. On gels containing glycol chitin, chitinase of C. violaceum strains showed four isoforms of 54-, 52-, 50- and 37-kDa, whereas there were four isoforms of 58-, 52-, 48- and 38-kDa in S. arcescens, three isoforms of 70-, 58- and 54-kDa in A. hydrophila and six isoforms of 90-, 79-, 71-, 63-, 58- and 38-kDa in A. caviae. The chitinase of C. violaceum strain C-61 was most active at pH 7.0 and at 5$0^{\circ}C$ and was stable in ranges of pH 5.0~10.0 for 2 hours and of 0~5$0^{\circ}C$ for 30 min.

• Journal of Life Science
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• v.11 no.2
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• pp.63-69
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• 2001

bacterium having high abilities to solubilize in-organic phosphate was isolated from cultivated soils. The strain was identified as Aeromonas hydrophila DA33, based on the physiological and biochemical properties. The optimum temperature and initial pH to solubilize insoluble phosphate in sucrose minimal medium were 3$0^{\circ}C$ and pH 5.0, respectively. In these conditions, phosphate-solubilizing activities of the strain against two types of insoluble phosphate were quantitatively determined. When glucose was used for carborn source, the strain had a marked mineral phospahte solubilizing activity. Inorganic phospahte solubilization was directly related to the pH drop by the strain. Analysis of the culture medium confirmed the production of gluconic acid as the main organic acid released by Aeromonas hydrophila DA33.

• Journal of fish pathology
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• v.1 no.1
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• pp.5-30
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• 1988

This is a comprehensive study for considering the effective treatment and control program of bacterial disease occurring in common carp, israel carp, color carp, crucian carp, eel and tilapia by clarifying the causes, mechanism of infection and onset and the diagnostic criteria. As a first step, the authors investigated the external views, gross and histopathologic findings of diseased fish using 450 infected fishes obtained from various farmer of Korea. This infection was characterized by hyperemia, hemorrhage and swelling of body surface and fins, congestion of liver, spleen, kidney, inflammation of intestine, hemorrhagic inflammation of various tissues, and necrosis and ulcer of various tissues were accompanied in serious cases. Bacteriologically, Aeromonas hydrophila and Edwardsiella tarda were isoiated from these fishes. Particularly in the regular check on 222 eels, 177 strains were isolated as 29.94% of Aeromonas hydrophila, 48.58% of Edwardsiella tarda and 21.47% of Flexibacter columnaris. Hexibacter columnaris was isolated from corroded gill of eels. The identical disease was occurred by innoculating the isolated Aeromonas hydrophila and Edwardsiella tarda and the identical strains were isolated from infected experimental fishes. The eels which were diagnosed Aeromonas disease from Kwangju, Pusan accompanied hemorrhage, swelling of body surface and fins, inflammation of stomach and intestine containing mucous fluids mixed with the pathogens. Color carp and crucian carp which were innoculated with the isolated 5 strins of Aeromomas hydrorphil died within 3 or 4 days accompanying with the characteristics of Aeromonas disease. Edward disease was characterized by abscesses of body surface, pus formation with concentration on phagocytes. The size of absecsses increased with progression elf disease. There were also various abscesses at internal organ and white nodules appeared in kidney. Histologically, various progressive granuloma were examined without inflammation of intestine. Columnaris disease of eels showed no hemorrhage except slight white body color. In autopsy, most of internal organs appeared normal and there were no septic odors. The only character was corrosion of gills. In order to treat these bacterial diseases, infected fishes must bathe in 20ppm chloramphenicol or kanamycin solution for 1 hour. Besides, medication program in oral ingestion of 75mg/kg chloramphenicol per day continuing for 5 to 7 days. After injecting the formalin treated Aermonas hydrophila antigen into carp, relatively high agglutination titer showed between 3 weeks and 6 weeks. Though this titer decreased from that time, it was continued for 18 weeks. In the case of injecting the formalin treated Edwardsiella tarda antigen into tilapia, the titer also increased. But tilapia which were immersed in the suspension fluid of the formalin treated Edwardsiella tarda showed no increase of the titer.

• Journal of fish pathology
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• v.5 no.2
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• pp.77-85
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• 1992

This study was carried out in order to investigate the immunosuppressive factor and immune response of color carp. The protection and serum antibody production of juvenile color carp aganist Aeromonas hydrophila were investigated on the effect of temperature differences and injected several aquatic drugs, i.e. Hydrocortisone, Oxytetracycline, Chloramphenicol and Ascorbic acid. The fish were injected intraperitoneally with 1mg/fish of HKC and FKC at three different temperature conditions as $16^{\circ}C$, $24^{\circ}C$, and $32^{\circ}C$ respectively. There were induced better protection and serum antibody production in the fish which had been kept at $24^{\circ}C$ than in the fish which had been kept at $16^{\circ}C$ and $32^{\circ}C$. The FKC immunized fish were followed 24 hrs later with intraperitoneal injection of 40mg/kg body weight of Hydrocortisone, 60mg/kg body weight of Oxytetracycline. 60mg/kg body weight of Chloramphenicol and 30mg/kg body weight of Ascorbic acid, respectively. The control fish were injected PBS only. The fish given the above aquatic drugs reduced serum antibody production level and protection rate when compared to control fish. As the results, immune response of juvenile color carp immunized FKC at $24^{\circ}C$ was more effective than $16^{\circ}C$ or $32^{\circ}C$ and immune response of juvenile color carp injected several aquatic drugs which was seemed to be immunosuppressive factor.

• Microbiology and Biotechnology Letters
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• v.44 no.2
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• pp.130-139
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• 2016

A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co²⁺, Cu²⁺, and Fe²⁺, but was recovered by metal chelating of EDTA. The K_m and V_max values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.522.3-523
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• 1986

A Screening test was carried out for chitin-decomposing bacteria. In 100 samples from soil, fesh water and sea water, 7 strains of Chitinolytic bacteria were isolated. 5-3K which exhibited the highest chitinase activity was identified as Aeromonas hydrophila and cultural conditions from maximum chitinase production were determined. Optimum Chitinase production was obtained at pH 7, 33eC and with chitin concentration greater tham 0.2% Under optimal conditions, high yields of Chitinase were obtained in 16-30 hours. Chitinase was purified by ammonium sulfate precipitation and sephadex G-100 gel-filtration from the culture filtrgte.

• Journal of fish pathology
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• v.30 no.2
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• pp.89-96
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• 2017

The majority of freshwater ornamental fish are imported and distributed domestically, causing high risk of exposure to exotic pathogens and drug resistant bacteria in Korea. Aeromonas hydrophila is known as a common species of fresh water bacteria and opportunistic fish pathogen, as well as a species causing zoonotic infection. In this study, we isolated motile aeromonads from various imported freshwater ornamental fish and studied the characters of the isolates. Imported freshwater ornamental fish were purchased on day 1 after the fish were deposited in the aquarium. Bacteria were isolated from the liver, kidney and spleen of fish using 0.5% NaCl containing tryptic soy agar medium. Bacteria were grouped on the basis of their morphological characteristics. The colonies with clear zone on starch-ampicillin agar (SA agar) were tentatively identified as Aeromonas spp. Two hundred and twenty-six strains, about 70% of total isolates were assumed to be Aeromonas spp. Nine isolates were further identified based on the result of the API 20E test and PCR using primers specific for A. hydrophila 16S rRNA gene. The isolates were identified as A. hydrophila and the API 20E test showed differences in trisodium citrate, D-sucrose, D-melibiose, amygdalin and L-arabinose availability between the nine isolates and standard A. hydrophila. The susceptibilities of the isolated bacteria to 10 antibacterial agents were confirmed by the disk diffusion method. Isolated strains were found to be resistant to amoxicillin and ampicillin and sensitive to florfenicol. However, 7 isolates showed multiple drug resistances to erythromycin, oxytetracycline, nalidixic acid etc. Pathogenicity of the isolates was determined by the artificial challenge test on goldfish (Carassius auratus). Three isolates caused 60 ~ 80% mortality in goldfish within 5 days after the initiation of challenge. These results indicate that multiple drug resistant, highly pathogenic and exotic A. hydrophila can spread to domestic aquarium and the preventive treatment of fish before sale is necessary.

• Journal of fish pathology
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• v.22 no.3
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• pp.391-400
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• 2009

Genetic identification of 17 fish-derived Aeromonas strains was attempted using 5 housekeeping genes. 16S rRNA, gyrB, rpoD, dnaJ and recA genes from the 17 strains were amplified, and total of 85 amplicons were sequenced. DNA sequences of the strains and type strains of the 17 Aeromonas homology groups were used for genetic identification and phylogenetic analyses. None of the strains was identified as a single species using the 16S rRNA gene, showing the same identities (average = 99.7%) with several Aeromonas species. According to gyrB, rpoD, dnaJ, and recA, 9 strains and RFAS-1 used in this study were identified as A. hydrophila and A. salmonicida, respectively. However, the other strains were closely related to 2 or more Aeromonas species (i.e., A. salmonicida, A. veronii, A. jandaei, A. media and A. troda) depending on the genetic marker used. In this study, gyrB, rpoD, dnaJ and recA gene sequences proved to be advantageous over 16S rRNA for the identification of field Aeromonas isolates obtained from fish. However, there are discrepancies between analyses of different phylogenetic markers, indicating there are still difficulties in genetic identification of the genus Aeromonas using the housekeeping genes used in this study. Advantages and disadvantages of each housekeeping gene should be taken into account when the gene is used for identification of Aeromonas species.