• Journal of Veterinary Clinics
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• v.27 no.1
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• pp.62-65
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• 2010

Arowana (Scleropages formosus) is the most valuable group of ornamental fishes and very much in demand in the ornamental fish trade and commands high price ranging from hundreds to thousands of dollars per fish. In this paper, we described a case of mortality of arowana from a private aquarium in Korea. A bacterial pathogen from fish organs (brain, kidney, liver) was cultured, identified and confirmed using Vitek System 2, API 20E test, multiplex PCR and 16S rRNA gene sequencing. The morphological and biochemical properties of the bacterium isolated from the brain, kidney and liver of the fish were similar to Aeromonas sobria. Positive amplification products using the multiplex PCR assay for detection of A. sobria were obtained from these organs. The 16S rRNA gene of the isolates from fish was identical and exhibited 100% sequence similarity with A. sobria (AY987762.1) strain available from GenBank. This bacterium contained hemolysin gene, a virulence factor that plays an important role in outbreaks of disease and is pathogenic to humans as well as in fish. Although this opportunistic bacterium was isolated from a fish without any external symptoms, this pathogen may act as a reservoir and enhance chances of zoonosis to human such as during handling.

• Korean Journal of Veterinary Research
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• v.30 no.2
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• pp.145-156
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• 1990

This study was purposed to produce blood typing reagents for classifing Cheju horse's blood group factors. The blood typing reagents were prepared by immunization of sixteen pairs out of fourty eight heads of Cheju horses, and seventeen different blood typing reagents (anti-$A_1$, A', H, Z, $ZZ_2$, C, J, K, $P_1$, Q, R, S, $T_1$, $N_1$, $U_2$, X and $E_2$) were prepared from equine isoimmunization. The result for the reagent production and the variation of agglutinin and hemolysin titer are as follows: 1. Anti-K, R, S, $N_1$, $U_2$ and X acted exclusively as hemolysins whereas anti-J, $P_1$, $T_1$ and $E_2$ acted exclusively as agglutinins, however, Anti-$A_1$, A', H, Z, $ZZ_2$, C and Q reacted both as hemolysins and agglutinins. 2. Among the antibodies in most of the reagents reacted both as hemolysins and agglutinins, But anti-$A_1$, H, Z, $ZZ_2$ and Q elevated higher agglutin titers than hemolysin titers, Anti-A' and anti-C elevated higher hemolysin titers than agglutinin titers. 3. Complex antibody $A_1U_2$ was adsorbed $U_2$ and unified to $A_1$, while complex antibody X $N_1$ went through an adsorption process and produced simple antibody X and $N_1$, respectively. 4. In the hemolytic reaction, anti-K and anti-R showed the highest titer, In the agglutination reaction, anti-Z and anti-$ZZ_2$ showed the highest titer. 5. In the case of No.138~No.140 horse immunization combination and No.133~No.146 horse immunization combination, Although production of complex antibody $CE_2$ and $JP_1$ were expected, only anti-$E_2$ and anti-J were produced respectively. 6. The degree of elevation of antibody titer was varied by the blood types and by the types of donor and recipient combination. The fast elevating types, slow elevating types and types that never elevate the antibody titer were identified.

• Korean Journal of Veterinary Research
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• v.50 no.4
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• pp.331-333
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• 2010

Moribund albino catfish, Clarias sp., displayed from an indoor private commercial aquarium were submitted in the laboratory for diagnostic examination. Dense culture of bacteria was recovered from the kidney and was characterized using Vitek System 2 and showed 98% probability to Aeromonas (A.) hydrophila. PCR result showed positive using A. hydrophila extracellular hemolysin gene ahh1 (130 bp) and aerolysin gene aerA (309 bp). The 16S rRNA gene was identical and exhibited 97% sequence similarity with the other known isolates of A. hydrophila available in the GenBank. In this paper, we reported the isolation and molecular detection of A. hydrophila from an albino catfish.

• Journal of Microbiology
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• v.38 no.1
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• pp.1-7
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• 2000

Eight strains of Aeromonas hydrophila isolated from diseased trout in Korea were characterized and compared with an American type strain by various methods including biochemical and physiological tests, PCR, randomly amplified polymorphic DNA (RAPD), plasmid profiling, and gel electrophoresis of total, membrane, and extracellular proteins. Virulence factors such as surface array proteins, cytotoxin, hemolysin, haemagglutinin, and protease were also investigated. The Korean strains showed heterogeneity in Iysine decarboxylase production, utilization of various carbon sources, and production of acetoin. Five strains had the same profiles of total and membrane proteins. Six strains haemagglutinated with trout red blood cells (RBCs) which was inhibited by fucose, galactose, and mannose, except for No. 1 where haemagglutination was inhibited by only galactose and mannose, but not by fucose. Four isolates haemagglutinated with human RBCs which was inhibited by fucose and mannose yet not by galactose. The type strain haemagglutinated only with trout RBCs which was inhibited by fucose, galactose, and mannose. Every isolate secreted protease, hemolysin, cytotoxin, and siderophore, but no enterotoxin. Results showed that the Korean isolates, except for No.7, had very different biochemical and molecular characteristics from those of the American type strain.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.49-55
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• 2000

Lysogenic conversion of Staphylococcus aureus to loss of ${\beta}-hemolysin$ production by serological group F phages is always associated with gain in staphylokinase production. In this study, the new phages belonging to serotype F were detected during the course of isolation of phages from Staph aureus of bovine origin and some characteristics of the new phages isolated were investigated. The new phages, ${\phi}470$ and ${\phi}499$, isolated from Staph aureus producing ${\beta}-hemolysin$ and staphylokinase(${\beta}^+\;K^+$) were found to convert ${\beta}^+\;K^+$ strain to ${\beta}^+K\;^+$, Staph aureus strains lysogenized by this serotype F single-converting phage ${\phi}470$ or ${\phi}499$ could be again lysogenized with serotype F double-converting phage ${\phi}506$. The frequency of lysogenization of indicator strains by serotype F single-converting phage was 100%, whereas the frequency for serotype F double-converting phage ${\phi}506$ varied from 4.2% to 97.6% according to the indicator strains. The indicator strain lysogenized with phage ${\phi}470$ was resistant to phage ${\phi}499$, and vice versa, but not to phage ${\phi}506$. Therefore, phage ${\phi}470$ and ${\phi}499$ were shown to be identical by immunity test.

• The Journal of Pediatrics of Korean Medicine
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• v.21 no.1
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• pp.189-209
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• 2007

Objectives : In order to study the effect of Dangkwiyughwangtang and Okbyoungpoongsangamibang on the immune response induced by methotrexate in mice. Method : Delayed type of hypersensitivity, hemagglutinin titer, hemolysin titer, rosette forming cells, phagocytic activity for immune response, lymphocyte transformation, and productivity of Interleukin-2 were measured. Results : Body weight decreasing was significantly inhibited as compared with control group in both the Dangkwiyughwangtang and Okbyoungpoongsangamibang groups. Delayed type of hypersensitivity was significantly increased as compared with control group in both groups Hemagglutinin titer was significantly increased as compared with control group in both groups. Hemolysin titer was significantly increased as compared with control group in the Okbyoungpoongsangamibang group. Rosette forming cells were significantly increased as compared with control group in both groups. Phagocytic activity for immune response was slightly decreased in the Dangkwiyughwangtang group and slightly increased in the Okbyoungpoongsangamibang group insignificantly as compared with the control group. Lymphocyte transformation was significantly increased as compared with the control group in both the Dangkwiyughwangtang and Okbyoungpoongsangamibang groups. Productivity of Interleukin-2 was significantly increased as compared with the control group in both the Dangkwiyughwangtang and Okbyoungpoongsangamibang groups. Conclusion : Both the Dangkwiyughwangtang and Okbyoungpoongsangamibang groups enhance immunity in mice.

• Journal of Food Hygiene and Safety
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• v.23 no.3
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• pp.248-256
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• 2008

To investigate the epidemiological characteristics of 68 Listeria monocytogenes isolates, including 11 reference strains and 57 isolates from imported US beef, domestic meats(beef, pork, chicken meat), raw milk, and milk plants. L. monocytogenes was to evaluate the production of virulence proteins, such as hemolysin(LLO) and lecithinase(LCP), the adsorption of Congo red(CRA), and to detect virulence genes using the polymerase chain reaction(PCR). In the study of virulence protein production, 68(100%), 62(91.2%), and 54(79.4%) of the 68 L. monocytogenes strains were positive for LLO production, the LCP test, and the CRA test, respectively, while strains of other species, such as L. innocua, L. gray, L. murrayi, and L. welshimeri, were not. There were no significant differences between L. monocytogenes serotypes and the ability to produce LLO or LCP. L. monocytogenesstrains had very high hemolytic titers(2 to 16 fold), while the other Listeria species, other than L. ivanovii and L. seeligeri, did not. The hemolysin activities of L. monocytogenes, L. ivanovii, and L. seeligeri usually exceeded 1.0 HU/mg, while those of other Listeria spp. were less than 0.04 HU/mg. In the PCR assay, all of the L. monocytogenes strains contained the hlyA, plcA, plcB, inlA, and inlB virulence genes and produced a product of the expected size. In the PCR of the actA gene, the expected 385-bp product was seen in 39(57.4%) L. monocytogenesstrains, while an unexpected 268-bp product was seen in 29(42.6%) strains. Most L. monocytogenes strains isolated from Hanwoo beef produced the 385-bp actA gene product, while strains of imported US beef usually produced the 268-bp actA gene product. By contrast, no virulence gene products were amplified in the other Listeria spp.

• Korean Journal of Food Science and Technology
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• v.37 no.1
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• pp.134-141
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• 2005

To characterize genotypic and phenotypic traits of Staphylococcus aureus isolates (n = 86) from lettuces and raw milk, major virulence-associated genes and antibiotic susceptibility were detected using PCR-based methods and disk diffusion method, respectively. All isolates possessed coagulase gene and showed five polymorphism types [500 bp (2.4%), 580 bp (17.4%), 660 bp (61.6%), 740 bp (17.4%), and 820 bp (1.2%)] due to variable numbers of tandem repeats present within the gene. Two or three different loci of hemolysin gene family were dominant in isolates, 47 of which (55%) possessed combination of hla/hld/hlg-2 genes as the most prevalent types. Among enterotoxin-encoding genes, sea was detected from 32 isolates (37%), sed from 1 isolate (1%), and sea and sed genes were co-detected from 4 isolates (5%), whereas seb, sec, and tsst-1 genes were not detected. All isolates were susceptible to ciprofloxacin, trimethoprim/sulfamethoxazole, oxacillin, and vancomycin, 85 isolates (99%) to penicillin G, 54 isolates (63%) to chloramphenicol, 51 isolates (59%) to erythromycin, and 7 isolates (8%) to clindamycin. Among resistant isolates, seven displayed multiantibiotic-resistance against two different antibiotics.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.168-176
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• 2005

Vibrio vulnificus, one of the marine bacterial pathogens causing septicemia, was detected using molecular methods, namely, PCR and/or Southern hybridization, and real-time PCR. Extracted and purified total DNAs by using commercial kits were used as templates for PCR. Multiplex-PCR was conducted by employing three sets of primers for the genes, hemolysin (vvhA), phosphomannomutase (pmm), and metalloprotease (vvpE), for V vulnificus virulence. The presence of DMSO ($5\%$) and BSA ($0.1\%$) in PCR reaction mixture improved a detection efficiency by higher PCR band intensities. TaqMan real-time PCR was carried out by using gene segment of vvhA as a target. Detection limit of PCR/Southern hybridization without enrichments was to be around $10^2\;cells\;g^{-1}$ of sample. However, those three methods using the enrichment at $35^{\circ}C$ in APW showed high sensitivity ($2\~10\;cells\;g^{-1}$ of sediments). Highly sensitive detection of V vulnificus by real-time PCR was achieved within $5\~6$ hr, whereas the detection by PCR/Southern hybridization required about 36 hr. Thus, it was evident that real-time PCR is the most rapid and efficient method for detecting V vulnificus in tidal flat sediments.

• Journal of Food Hygiene and Safety
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• v.12 no.1
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• pp.9-14
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• 1997

Vibrio mimicus is a closely related species with V. cholerae, and has been reported to be associated with gastrointestinal infections. Although extraintestinal infections of these vibrios have also been reported in Japan and Southeast Asia. But little research papers on V. mimicus was reported in Korea. Therefore, we tried to isolate V. mimicus from the environmental sea water from April to July in Pusan, Korea. Among the isolated strains, we selected the strongest hemolytic strain and then named V. mimicus SM-9. In this paper, we checked the antibiotic susceptibility and psychrotrophic characteristics of the isolated strain. Hemolytic activity of the hemolysin produced by the isolated strain was also measured. V. mimicus was not detected from the sea water samples in April and May, but its detection rate was relatively high in June and July in Pusan, Korea. The bacteriological characteristics of V. mimicus SM-9 were Gram-negative rods, motile, oxidase positive, Voges-Proskauer negative and sucrose negative. In 23 kinds of antibiotics susceptibility test, V. mimicus SM-9 showed susceptibility to the most of antibiotics submitted while it was resistive against lincomycin, oxacillin, rifampin and vancomycin. Hemolytic activity of the hemolysin produced by V. mimicus SM-9 was highest in stationary phase of the growth curve in BHI broth at 37$^{\circ}C$ and its activity was reached 18 HU per $m\ell$ of culture supernatant. For checking the psychrotrophic property of V. mimicus SM-9, the decreasing rate of the strain in phosphate buffer solution and yellowtail flesh homogenate was examined during the storage at 4, 0, -4 and -2$0^{\circ}C$. The decreasing rates of the selected strain stored in phosphate buffer solution were greater than those in fish homogenate. Decreasing rates of V. mimicus SM-9 stored in phosphate buffer solution were not significantly different by the storage temperatures. The viable cell counts of the strain were decreased as 5 log cycles after 120 hours at all the tested temperatures. While decreasing numbers of the strain in fish homogenates were 2*4 log cycles after 120 hours. The decreasing pattern of the strain numbers were very slow after 200 hours at all the stored temperatures.