• 대한임상검사과학회지
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• 제46권4호
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• pp.140-142
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• 2014

To evaluate the recovery rates to increase toxigenic C. difficile, the selective enrichment broth culture methods were compared with commonly used cytotoxin assays and toxigenic culture. First, the enrichment culture, using the selective medium broth for 2 to 5 days, was performed and then, toxigenic C. difficile was confirmed by C. difficile toxin gene-specific PCR after being cultured on C. difficile selective agar. The sensitivity of C. difficile from the enrichment culture (100%) was higher than that of C. difficile selective agar culture (93.8%), while positive predictive values (PPV) were low; 72.7% (16/22) and 88.2% (15/17). PPV of the enrichment culture are not high. Recently, combinations of C. difficile selective agar culture, C. difficile A & B assays, glutamate dehydrogenase, and nucleic acid amplification method are widely used. The enrichment culture was disadvantageous in PPV, turn-around time, and cost. So, what we performed is not considered as a common method of diagnosis of C. difficile-associated diarrhea.

• Mycobiology
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• 제31권4호
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• pp.183-190
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• 2003

Five cereal grains(wheat, barley, rice, maize and sorghum) were collected from three Egyptian provinces known to be grain producers(Daqahlia, Gharbia and Kafer el-Sheikh). Two species of Alternaria(A. raphani and A. tenusinae); two species of Aspergillus(A. flavus and A. niger); one species of Cunninghamella(C. elegans); one Dreschslera species(D. myakt); three Fusarium species(F. graminearum, F. moniliform and F. solani); one Rhizopus species(R. stolonifer) and two species of Penicillium(P. digitatum and P. notatum) were isolated from the grains. The densities of these fungi and their frequencies of occurrence have been investigated. All the fungal isolates were tested for the production of toxic metabolites in culture media and the percentages of toxigenic isolates were calculated. The biological assay of the toxigenic fungal isolates showed significant variations in toxigenic activity. Thin layer chromatography revealed that the most active isolate produces moniliformin in culture media. The effect of culture conditions on the production of moniliformin was studied.

• 대한수의학회지
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• 제40권1호
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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.

• 한국식물병리학회지
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• 제10권2호
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• pp.129-135
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• 1994

Ninety-two isolates of Fusarium species were obtained from Chinese corn samples. The predominant Fusarium species isolated from corn seeds were F. moniliforme, F. proliferatum, F. oxysporum and F. subglutinans, and all 13 species were identified. Each isolate was grown on autoclaved wheat grains and wheat cultures were fed by twenty-one-day-old female rats for the toxicity test. Twenty-six out of 92 isolates caused the death accompanying feed refusal, severe weight loss, liver damage, and hemorrhages in the stomach and intestines. Of the toxigenic isolates, 17 isolates of F. moniliforme, 4 of F. oxysporum, 3 of F. proliferatum, and one of each F. sporotrichioides and unknown species were lethal to rats. The analyses of fumonisin B1 production of the 26 toxigenic Fusarium isolates were carried out by thin layer chromatography and high-performance liquid chromatography, and fumonisin B1 was confirmed by mass spectrometry. Fumonisin B1 was produced in wheat culture at levels ranging from 280 $\mu\textrm{g}$/g to 3,952 $\mu\textrm{g}$/g by all of toxigenic F. moniliforme and F. proliferatum, but by none of the other toxigenic Fusarium species. The present results suggest the high possibility of natural occurrence of fumonisin B1 in corn samples imported from China.

• Mycobiology
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• 제44권3호
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• pp.155-161
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• 2016

The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through ${\alpha}-amylase$ and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in ${\alpha}-amylase$ and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that ${\alpha}-amylase$ activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.

• Journal of Microbiology
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• 제44권3호
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• pp.320-326
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• 2006

Pasteurella multocida is an important veterinary and opportunistic human pathogen. In particular, strains of P. multocida serogroup D cause progressive atrophic rhinitis, and produce a potent, intracellular, mitogenic toxin known as P. multocida toxin (PMT), which is encoded by the toxA gene. To further investigate the toxigenic and pathogenic effects of PMT, a toxA-deleted mutant was developed by homologous gene recombination. When administrated to mice, the toxigenicity of the toxA mutant P. multocida was drastically reduced, suggesting that the PMT constributes the major part of the toxigenicity of P, multocida. Similar results were obtained in a subsequent experiment, while high mortalities were observed when toxA(+) P. multocida bacterial culture or culture Iysate were administrated. Mice immunized with toxA(-) P. multocida were not protected (none survived) following challenge with toxA(+) P. multocida or bacterial culture Iysate (toxin). These results suggest that the toxigenicity of P. multocida is mainly derived from PMT.

• 대한수의학회지
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• 제44권4호
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• pp.551-559
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• 2004

Shiga toxin-producing Escherichia coli (STEC) causes various clinical signs in human and animals, and has been indicated as a global enteropathogen with zoonotic importance. In this study, the feces of healthy pigs were collected from the slaughtered pigs of Daejon abattoir during the period from December 2001 to October 2002. Of 326 specimens, 13 STEC were confirmed by culture, PCR and colony hybridization. The isolates were further studied for toxin types, pathogenic factors, plasmid profiles, and antimicrobial resistance to characterize the genetic and toxigenic properties. In PCR, all of 13 isolates were evident to have shiga toxin gene (stx). Of 13 isolates stx1 gene was detected in 4 and stx2 gene in 9. The genes of eaeA, hlyA and rfbE were not present in any isolates. In colony hybridization using shiga toxin common primer (STXc), 2 to 9 per 100 colonies subcultured from 13 isolates showed the positive reaction. In the examination for plasmid profiles of the isolates, one to eleven plasmids with varying sizes of 1.0 Kb to 100 Kb were detected, and the 13 STEC could be classified into four groups by the plasmid patterns. The antimicrobial resistance patterns of the isolates were comparably corresponded with the plasmid profile patterns.

• 한국균학회지
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• 제1권1호
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• pp.17-21
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• 1973

Fifty-eight strains of Aspergillus spp. isolated from various grains were examined for the screening of aflatoxins by the method of the Thin-Layer Chromatography (TLC), using the aflatoxin producing strain of Aspergillus flavus ATCC 15517 as a control. The results as follows: 1. Samples of Aspergillus spp. isolated from various grains were extracted with chloroform and chromatographied by the thin-layer chromatography method. 2. Three strains of Aspergillus spp. among the 58 strains can be found that the spots having the same Rf value as control with culture extract of Aspergillus flavus ATCC 15517. 3. The further prove studies of aflatoxins were proceed by the methods of in vivo and in vitro tests. And this methods considered to capable for the use of mass screening among the samples.

• 대한미생물학회지
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• 제8권1호
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• pp.43-52
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• 1973

Twenty culture filtrates among the various isolated strains from foodstuffs were submitted for toxicity screening using HeLa cells and mice. Fourteen strains(70%) were toxic to both HeLa cells and mice, 17 strains(85%) to HeLa cell alone and 14 strains(70%) to mice alone. As a mass screening this method employed is feasible to detect mycotoxin-producing fungi. In most instances, the results obtained by HeLa cells were in good parellelism with those obtained by mice.

• 한국환경보건학회지
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• 제35권2호
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• pp.95-99
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• 2009

This study was performed to investigate the possible effect of garlic produced in Korea on the inhibition/reduction of growth of A. parasiticus, a toxigenic strain. The effect was studied using different concentrations of freeze-dried garlic in potato-dextrose agar (PDA) and/or in yeast-extract sucrose (YES) broth at $25^{\circ}C$ for 15 days. While inhibition of the fungal growth due to increasing the concentration of garlic was observed, the more remarkable effect was observed on the ninth day. Reduction of fungal diameter as a result of addition of garlic on PDA was observed to range between 3.4% to 20.1 % while reduction of mycelial weight in YES broth ranged from 9.9% to 30.5%. The 0.5% and 1.0% concentrations of garlic significantly reduced fungal diameter in PDA on the 9th day, while 0.1 %, 0.5%, and 1.0% concentrations of garlic significantly reduced the mycelial weight in YES broth (p<0.05). Dose-response relationships were observed between the concentration of garlic and inhibition of growth both in solid culture and in liquid culture. This study indicates that garlic could be an effective inhibitor at a human consumption level of the growth of A. parasiticus. More research is needed to study the inhibitory effects of the main active component of garlic.