• Korean Journal Plant Pathology
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• v.14 no.2
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• pp.177-183
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• 1998

A DNA fragment which is involved in tolassin production was cloned to obtain a molecular marker of Pseudomonas tolaasii, a casual agent of bacterial brown blotch disease of oyster mushroom (Pleurotus ostreatus). Tolaasin is a lipodepsipeptide toxin and known as a primary disease determinant of the P. tolaasii. It is responsible for formation of white line in agar when P. tolaasii were cultured against white line reacting organisms (WLROs). White line negative mutants (WL-) were generated by conjugation between rifampicin resistant strain of P. tolaasii and E. coli carrying suicidal plasmid pSUP2021 : : Tn5. The ability of tolaasin production of the WL- mutants was examined by hemolysis test, pathogenicity test, and high pressure liquid chromatography (HPLC) analysis of culture filtrate. All of the WL- mutants were lost the ability of tolaasin production (Tol-). Genomic library of the Tol- mutant was constructed in pLAFR3 and the cosmid clone containing Tn5 was selected. DNA fragment fro franking region of Tn5 was cloned from the plasmid and used as a probe in Southern blot. DNA-DNA hybridization with the probe to total DNA from group of bacteria ecologically similar to P. tolaasii including WLORs, fluorescent Pseudomonads isolated from oyster mushroom, P. agarici, P. gingeri, and some of other species of Psedomonas showed that some of the tested bacteria do not have any hybridized band and others have bands sowing RFLP. The cloned DNA fragment or its nucleotide sequence will be useful in detection and identification of the P. tolaasii.

• Journal of Preventive Veterinary Medicine
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• v.42 no.4
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• pp.148-156
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• 2018

Clostridium perfringens (C. perfringens) may cause diarrhea and enterotoxemia in adult and young livestock, leading to problems in the production and management of farms. Four hundred fecal samples were collected from 25 goat farms located in Gyeongsangbuk-do Province in the Republic of Korea. Sixteen C. perfringens strains were isolates from fecal samples, and the isolates were identified as type A (n=11) and type D (n=5). Additionally, ${\alpha}$- and ${\varepsilon}$-toxin genes were detected in 16 and 5 strains by PCR, respectively, and the enterotoxin gene was presented in 2 strains. The antibiotic susceptibility test was performed using the disk diffusion method and E-test method. In the disk diffusion method, ampicillin (n=16) and chloramphenicol (n=15) were highly susceptible to 16 C. perfringens isolates. In the E-test method, ampicillin, amoxicillin, amoxicillin/clavulanic acid and meropenem were susceptible to more than 14 of 16 C. perfringens isolates. This study indicates that administration of antibiotics such as ampicillin, amoxicillin/clavulanic acid and meropenem can prevent and treat C. perfringens infections in goats.

• Mycobiology
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• v.44 no.1
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• pp.38-47
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• 2016

The ascomycete fungus Mycosphaerella graminicola (synonym Zymoseptoria tritici) is an important pathogen of wheat causing economically significant losses. The primary nutritional mode of this fungus is thought to be hemibiotrophic. This pathogenic lifestyle is associated with an early biotrophic stage of nutrient uptake followed by a necrotrophic stage aided possibly by production of a toxin or reactive oxygen species (ROS). In many other fungi, the genes CREA and AREA are important during the biotrophic stage of infection, while the NOXa gene product is important during necrotrophic growth. To test the hypothesis that these genes are important for pathogenicity of M. graminicola, we employed an over-expression strategy for the selected target genes CREA, AREA, and NOXa, which might function as regulators of nutrient acquisition or ROS generation. Increased expressions of CREA, AREA, and NOXa in M. graminicola were confirmed via quantitative real-time PCR and strains were subsequently assayed for pathogenicity. Among them, the NOXa over-expression strain, NO2, resulted in significantly increased virulence. Moreover, instead of the usual filamentous growth, we observed a predominance of yeast-like growth of NO2 which was correlated with ROS production. Our data indicate that ROS generation via NOXa is important to pathogenicity as well as development in M. graminicola.

• Korean Journal of Microbiology
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• v.9 no.2
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• pp.86-93
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• 1971

Escherichiae-like organisms were isolated from rectal specimens of 56 children who were either in preschool age or in elementary school. The isolated strains were subjected to tests to screen enteropathogens producing heat-labile enterotoxin and susceptibility test to various antibiotics by disc diffusion method on agar plates. Production of heat-labile enterotoxin by the strains was assyed in the sensitive and reproducible cultured adrenal tumor cell system. The assay was sterodogenesis of the cell in the presence of heat-labile enterotoxin. Among 56 strains, gave positive reaction in the test of toxin production. This meant that about 10% of the children population objected to the study harbored the toxigenic strain of enteropathogenes. Some of these toxigenic strains were resistant to the antibiotics employed in the test. This study suggested that considerable population in Korea may harbor entertoxigenic E. coli as a part of intestinal normal flora. The toxigenic strains which are resistant to antibiotics may bring issue of public health in the future.

• Korean Journal of Veterinary Research
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• v.40 no.1
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• pp.56-62
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• 2000

Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.

• The Journal of the Korean Society for Microbiology
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• v.22 no.4
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• pp.377-392
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• 1987

Monoclonal antibody to the Escherichia coli heat-stable enterotoxin(ST) was produced to develop a rapid and convenient diagnostic method to the ST. The toxin was purified from culture supernatant of enterotoxigenic E. coli O148H28($ST^+/LT^+$) and conjugated to bovine serum albumin(BSA). The ST-BSA conjugate was used to immunize BALB/c mice and the immune spleen cells from these mice were fused with $P3{\times}63$ Ag8.V653 plasmacytoma cells. Hybridomas were screened by ELISA and positive hybridomas were cloned by limiting dilution. Finally, one stable clone (AS36) specific to ST was selected for further growth and characterization. Antibody titers of culture supernatant and ascitic fluid from BALB/c mice were 1:1,024 and 1:20,480 respectively in ELISA. The isotype and subclass of monoclonal antibody was IgG1 in sandwich ELISA. To test the neutralizing effect of monoclonal antibody on toxin activity of ST, mixture of ascitic fluid and ST was assayed by infant mouse assay and this monoclonel antibody was proved to be a neutralizing antibody. The titer of ascitic fluid which completely neutralized biological activity of 4 units of ST was 1:4. Purified ST was quantitatively measured by competitive ELISA and minimum amount of ST detectable by this assay was 250pg, which was an amount six-fold smaller than that detectable by infant mouse assay. Four reference strains of enterotoxigenic E. coli from WHO were detected by competitive ELISA and highly specific, sensitive and reproducible result was obtained.

• The Plant Pathology Journal
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• v.39 no.1
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• pp.88-107
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• 2023

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7℃, indicating an INA⁺ bacterium. The ice-nucleating temperature was -4.7℃ for a non-treated control (sterilized distilled water), whereas it was -9.6℃ for an INA^- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

• The Plant Pathology Journal
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• v.16 no.3
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• pp.137-141
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• 2000

A causal fungus of turfgrass blight was isolated from the infected leaves of zoysiagrass (Zoysia japonica Steud.) and identified as Nigrospora sphaerica (Sacc.) Mason by using a light misroscope. Its conidia are large (14-20 ${\mu}{\textrm}{m}$ diameter), shiny, black, aseptate, and smooth-walled spheres. The fungus caused typical blighting symptoms on the two turfgrass plants of bermudagrass (Cynodon dactylon (L.) Pers.) and bentgrass (Agrostis palustris Huds.). The fungus was found to produce a phytotoxic subtance to be associated with the pathogenic mechanism. A phytotoxin was isolated from the liquid cultures of N. sphaerica by repeated silica gel column chromatography and its structure was determined to be 5, 6-dihydro-5-hydroxy-6-propenyl-2H-pyr-2-one (T-3 compound). It was not a host-specific toxin showing phytotoxic effects to various plants inclusing turfgrasses in the leaf-wounding assay, the whole plant test, and the cellular leakage test. The compound caused leaf tip dieback symptoms in turfgrass plants similar to those caused by the pathogen. Thus, T-3 compound is thought to be involved in the development of Nigrospora blight.

• Korean Journal of Veterinary Service
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• v.19 no.2
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• pp.139-153
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• 1996

Porcine E coli infection is a disease caused by Enterotoxin produced by Enterotoxigenic Escherichia coli(ETEC). Enteric colibacillosis has become an economically important disease in pigs as a result of increasing intensification of farrowing management. The present study undertaken to obtain the antibiotic sensitivity and distribution of serogroups and pili producibility test of ETEC from E. coli isolates in Chonnam. The results obtained were as follows. 1. A total of 71 isolates identified as E, coli employing IMViC system from rectal specimens of 54 piglets with diarrhea. 2. In antibiotic sensitivity test, isolates showed high sensitivity to AN, CM, Fox, GM, but resistance to EM, NA TC. 3. The distribution of 25 Isolates of serogroups were 0141:K85(11.3%), 08:K87(8.5%), 064:K (5.6%), 0138:K8l (4.2%), 0139 :K82(2.8%), 0157:K88ac(1.4%) and 0149:K9l (1.4%). 4. MRHA of guinea pig erythrocytes was detected in 8 out of 25OK serotypes and 9 out of 46 unidentified serotypes. MRHA titers of serotypes showed from 64 to 128 in 0141: K85, 2 in 0138:K8l and no titers in 0139:K82. 5. The production of heat labile enterotoxin of ETEC was detect 39 out of 52 isolates showed $\beta$-hemolysin, 7 out of 52 isolates showed ${\gamma}$-hemolysin and 6 out of 52 isolates showed ${\gamma}$-hemolysin by $GM_1$ganglioside ELISA. The distribution of LT toxin were in 12 isolate showed $\beta$-hemolysin, 2 isolates showed ${\alpha}$-hemolysin and 3 isolates showed ${\gamma}$-hemolysin in 25 OK serotypes.

• KSBB Journal
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• v.15 no.1
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• pp.100-105
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• 2000

Optimal medium composition, culture conditions, characteristics of phase variation and activity of insecticidal toxin by Xenorhabdus nematophilus isolated and identified from Korean entomopathogenic nematode Steinernema carpocapsae were examined. Optimal medium composition of this strain was 50-70 g/L yeast extract, 3 g/L $K_{2}HPO_{4}$, 1g/L $NH_{4}H_{2}PO_{4}$, 2g/L ${MgSO}_4$$\cdot$${7H}_{2}O$, 10g/L NaCl and, these, yeast extract was found as a limiting nutrient for cell growth. When Monod equation was applied, maxmum specific growth rate and Monod constant were estimated as 0.13 $hr^{-1}$ and 20g/L, respectively. The pH of culture medium increased up to 8.5-9.5 regardless of initial pH 6-7 as the cells continued to grow. The specific growth rate in a 7 L fermentor was 0.18 $hr^{-1}$, which was enhancement 1.4 fold compared to a flask culture. In case of phase variation, phase I fraction was maintained above 90% at the stationary phase for both flask and fermentor cultures. According to oral toxicity test of Gallena mellonella by Xenorhabdus nematophilus, the addition of cell pellets into feed inhibited normal growth of insect larvae and killed completely then after 20 days cultivation. When culture supernatant of this strain was injected into hemolymph of insect larva, the toxicity was strongest at 24hr cultivation in the early exponential phase and gradually decreased as the culture time proceeded.