• Research in Plant Disease
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• v.11 no.2
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• pp.128-134
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• 2005

This study was conducted to elucidate effect of environmental factors on the development of white rot. In order to identify the causal agents causing white rot of Allium crops, we compared DNA profiles of a representative isolate, Sclerotium cepivorum, introduced from foreign country with Korean isolates using UP-PCR. As a result, Sclerotium isolates forming round-shaped sclerotia were identified as Sclerotium cepivorum pertaining in UP-PCR b group and Sclerotium isolates farming anamorphic-shaped sclerotia presumed to be a novel species of Sclerotium based on DNA profiles of UP-PCR. There was a big difference in DNA band pattern between two species of Sclerotium isolated in Korea. Electron micrographs of scanning electron microscope and transmission electron microscope showed morphological differences in sclerotial surface structure and rind layers between two species of Sclerotium. There were more wrinkles and pore spaces on sclerotial surface of Sclerotium sp. forming anamorphic-shaped sclerotia than that of Sclerotium cepivorum forming round-shaped sclerotia. Both of two white rot pathogens grew well at the temperature range of $10-25^{\circ}C$ with optimal temperature of $20^{\circ}C$. Sclerotia of the two pathogens were well formed at $20^{\circ}C$ and well germinated at the temperature range of $20-24^{\circ}C$, Effect of pre-incubation of sclerotia on destruction of sclerotial dormancy of two pathogens was evaluated through storing sclerotia under different temperature condition. The sclerotia of the two pathogens showed an increased capacity to germinate on potato dextroise agar when the sclerotia were incubated for 7 days at $10^{\circ}C$ after pre-treatment at $35^{\circ}C$ for 7 days. At that time, germination rate of Sclerotium sp. and 5. cepivorum was $100\%\;and\;70\%$, respectively. Flooding period and treatment temperature had an effect on sclerotial survival rate of the two pathogens. As flooding period and treatment temperature increased, sclerotial germination rate of the two pathogens decreased. It was confirmed that soil humidity played an important role on development of white rot. It was the highest disease incidence of garlic white rot when garlic were sown at potted soils infested with the two pathogens and adjusted soil humidity to $15\%$ (field moisture capacity, about -300 mb). As soil humidity increase or decrease based on $15\%$ of soil humidity, disease incidence decreased move and more.

• Annals of Clinical Microbiology
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• v.22 no.1
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• pp.9-13
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• 2019

Background: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.

• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.407-412
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• 2017

The aim of this study was to shorten the time required for subculture and bacterial identification and obtain a simple and rapid identification method for new test methods for bloodstream infections. The following results were obtained using a mass spectrometer. In Vitek 2, 208 (81.8%) cases were well-identified and 45 isolates were not identified in blood cultures. Among 208 cases, 146 (57.5%) were Gram positive bacteria and 108 (42.5%) were Gram negative bacteria. In total, 233 were identified to the species level and 21 were identified to the genus level. The identification error was found to be Propionibacterium acnes as Clostridium bifermentans. The accuracy of Enterobacteriaceae, glucose non-fermentative bacilli (GNFB), and staphylococci were 81/83 (97.6%), 12/15 (80.0%), and 72/85 (84.7%), respectively. The concordance rate of Vitek 2 and Vitek MS by the direct method was 81.8% and 45 isolates were not identified. Most of the unidentified bacteria were Gram positive bacteria (N=37). The Gram positive bacteria were streptococci (14), coagulase-negative staphylococci (CNS) (11), enterococci (3), Staphylococcus aureus (2), Micrococcus spp. (2), Bacillus spp. (2) and Actinomyces odontolyticus, Finegoldia magna, and Peptostreptococcus spp. The results reporting time was reduced to 24~72 hours compared to the conventional method. The rate of identification of the aerobic and anaerobic cultures was similar, but the use of an anaerobic culture did not require a dissolution process, which could shorten the sample preparation time. These results suggest that the method of direct identification in blood cultures is very useful for the treatment of patients. In further studies, it might be necessary to further improve the method for identifying streptococci and CNS, which were lacking in accuracy in this study.

• The Journal of the Korean Society for Microbiology
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• v.9 no.1
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• pp.13-18
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• 1974

Blood is one of the most important clinical specimens for the isolation of bacteria. A rapid isolation and a high isolation rate of bacteria are very important in blood culture because bacteremic patients are mostly in grave condition. Various blood culture media which support growth of most fastidious bacteria are available commercially. However, growth of bacteria are frequently delayed because of antibacterial activity of blood. Sodium polyanethol sulfonate(Liquoid) has been reported to inactivate the antibacterial substance and disrupt phagocytic cells. The beneficial effect of SPS is well recognized in the isolation of gram-positive bacteria. However, the effect does not seem to be prominent for gram-negative bacilli isolation mainly due to the rapidity of their growth. It has been experienced with Sal. typhi that the growth is much slower than that of other gram-negative bacilli. For the rapid growth of the organism, use of bile broth has been recommended. Although Sal. typhi is the most frequently isolated organism at present, about one half of total isolates are other organisms and, in case bile broth is used, other media which support growth of these organisms should be used together. Fluid thioglycollate medium(FTM) which is always used in blood culture to isolate anaerobes is inferior to brain heart infusion(BHI) for the isolation of aerobes. This study was done to determine the effect of SPS on the isolation of Sal. typhi from blood. During the Sep. 1973 to Sep. 1974 study period, 2460 blood cultures were made from the Severance hospital patients: BHI and FTM sets 1431 specimens, BHI with SPS(0.05%) and FTM sets 396 specimens, BHI and FTM with SPS sets 359 specimens, BHI and BHI with SPS sets 274 specimens. Mean incubation time required for the macroscopic detection of growth of Sal. typhi were 3.5 days on BHI and 2.7 days on BHI with SPS. The 0.8 day difference was statistically significant. On FTM the mean incubation time was 3.8 days while it was 2.9 days on FTM with SPS. The 0.9 day difference was statistically significant. The result on BHI with and without SPS sets showed faster growth on BRI with SPS in 7 specimens and slower growth in one specimen and the remaining 12 showed growth at the same time. These specimens had mean incubation time of 3.2 days on BHI and 2.3 days on BHI with SPS. The 0.9 day difference was statistically significant. This study indicates beneficial effect of SPS for the rapid isolation of Sal. typhi from clinical blood specimens.

• Korean Journal of Clinical Laboratory Science
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• v.38 no.3
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• pp.166-172
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• 2006

Metallo-${\beta}$-lactamase (MBL) can hydrolyze all ${\beta}$-lactams except monobactams and frequently coexists with various antibiotic resistance genes such as aminoglycoside resistance, sulfonamide resistance gene, etc. Therefore, the effective antibiotics against infections by these bacteria are markedly limited or can't even be found. We tried to search in-vitro antimicrobial combinations with synergistic effects for a VIM-2 type MBL producing Pseudomonas aeruginosa, isolated from clinical specimen. On the selection of antibiotic combinations with synergistic effects, we performed a one disk synergy test, modified Pestel's method, in agar without aztreonam (AZT). The bacteriostatic synergistic effects of this tests were scored as $S_1$ (by susceptibility pattern in agar without antibiotics), $S_2$ (by the change of susceptibility in agar with or without antibiotics) and $S_3$ ($S_1$ + $S_2$) and was classified into weak (1 point), moderate (2 points) and strong (3 points) by $S_3$ score. Subsequently, we carried out the time-killing curve for the antibiotic combinations with the strong synergistic bacteriostatic effect. One VIM-2 type MBL producing P. aeruginosa confirmed by the PCR showed all resistance against all ${\beta}$-lactams except AZT, aminoglycoside and ciprofloxacin. In the one disk synergy test, this isolate showed a strong bacteriostatic synergistic effect for the antibiotic combination of AZT and piperacillin-tazobactam (PIP-TZP) or AZT and amikacin (AN). On the time-killing curve after six hours of incubation, the colony forming units (CFUs/mL) of this bacteria in the medium broth with both combination antibiotics were decreased to 1/18.7, 1/17.1 of the least CFUs of each single antibiotics. The triple antibiotic combination therapy including AZT, PIP-TZP and AN was shown to be significantly synergistic after 8 hrs of exposure. In a VIM-2 MBL producing P. aeruginosa with susceptibility for AZT, the triple antibiotic combination therapy including AZT, PIP-TZP and AN may be considered as an alternative antibiotics modality against the infection by some MBL type. But the antimicrobial combination therapy for many more MBL producing isolates is essential to know as soon as possible for the selection of effective treatment against the infection by this bacteria.

• Journal of Ginseng Research
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• v.38 no.2
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• pp.136-145
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• 2014

Background: This study aimed to develop a biocontrol system for ginseng root rot caused by Fusarium cf. incarnatum. Methods: In total, 392 bacteria isolated from ginseng roots and various soils were screened for their antifungal activity against the fungal pathogen, and a bacterial isolate (B2-5) was selected as a promising candidate for the biocontrol because of the strong antagonistic activity of the bacterial cell suspension and culture filtrate against pathogen. Results: The bacterial isolate B2-5 displayed an enhanced inhibitory activity against the pathogen mycelial growth with a temperature increase to $25^{\circ}C$, produced no pectinase (related to root rotting) an no critical rot symptoms at low [$10^6$ colony-forming units (CFU)/mL] and high ($10^8CFU/mL$) inoculum concentrations. In pot experiments, pretreatment with the bacterial isolate in the presumed optimal time for disease control reduced disease severity significantly with a higher control efficacy at an inoculum concentration of $10^6CFU/mL$ than at $10^8CFU/mL$. The establishment and colonization ability of the bacterial isolates on the ginseng rhizosphere appeared to be higher when both the bacterial isolate and the pathogen were coinoculated than when the bacterial isolate was inoculated alone, suggesting its target-oriented biocontrol activity against the pathogen. Scanning electron microscopy showed that the pathogen hyphae were twisted and shriveled by the bacterial treatment, which may be a symptom of direct damage by antifungal substances. Conclusion: All of these results suggest that the bacterial isolate has good potential as a microbial agent for the biocontrol of the ginseng root rot caused by F. cf. incarnatum.

• Korean journal of applied entomology
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• v.15 no.3 s.28
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• pp.153-159
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• 1976

Present paper was attempted to investigate the effects of the light on the sclerotium formation of some isolates of Sclerotinia sclerotiorum(Lib.) do Bary from lettuce, cucumber and rape. The investigations were performed in the both aspects inducing sclerotial primordia from the mycelial colony and developing such primordia. The cultures were grown on potato-dextrose agar and were treated with day-light fluorescent lamps. In the continuous light illumination for 21 days, numbers of sclerotial primordia and matured sclerotia were increased with increasing the light intensity up to about 500 Lux, but on the contrary the dry-weight of matured sclerotia was decreased. In the 800 Lux illumination, induction of the primordia was extremely depressed, and then matured sclerotia were almost not produced. Short-time illumination for 48 hours, although the light intensity was as high as 5000 Lux, increased the numbers of sclerotial primordia and matured sclerotia. Dry-weight of matured sclerotia, however, was slightly increased only at low intensity of 160 Lux. On the other hand, the light shock which u·as alternatively on-of light every one minute for 48 hours increased the number and dry-weight of sclerotia produced, but the former was more effective at 500 Lux shock and the later more effective at 160 Lux shock.

• KSBB Journal
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• v.17 no.3
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• pp.276-282
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• 2002

Symbiotic bacteria with highly effective insecticidal activities were isolated and compared with their physiological characteristics from seven species of entomopathogenic nematodes belong to Steinernamatidae and Heterorhabditidae sp., and three of them were identified as Xenorhabdus nematophilus. Culture characteristics, insecticidal activities, pretense activities and fatty acid contents of various symbiotic bactierial isolates were also examined. In the case of cell growth and insecticidal activity, XR-PC and XR-MK were superior to other species when cultured in vitro. The insecticidal activity were highest at the early exponential growth phase, and gradually decreased with time. The protease activity of XR-DR was remarkable compared to other species. In the case of HE-HY, however the pretense activity increased in parallel with cell growth. Interestingly, the fatty acid patterns of Xenorhabdus nematophilus isolated from different emtomopathogenic nematode, showed remarkable differences in their contents of 12:0, 14:0, 16:1 cia 5 and 17:0 cyclo and hydroxy and branch factty acids were varied from 2% to 15% among total fatty acid contents.

• Food Science of Animal Resources
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• v.29 no.4
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• pp.533-538
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• 2009

To suggest an improved diagnostic method for Salmonella spp., analyses were conducted with immunoliposomes and compared with the results from a commercial test kit. One sample out of 36 samples of eggshell was Salmonella-positive via immunoliposomes. In the case of the use of the commercial test kit, six samples out of 36 samples were Salmonellapositive. These Salmonella-positive samples were subjected to biochemical identification tests that confirmed that they were Salmonella-negative. As for the egg content samples, they were Salmonella-negative in both analyses with immunoliposomes and the commercial test kit. The Salmonella analysis with immunoliposomes reduced detection time, by 24 h compared to the commercial test kit. Bacteria, including Acinetobacter baumanni, Chryseomonas luteola, Enterobacter cloacae, Escherichia coli, Escherichia hermannii, Klebsiella pneumonia, Pantoea spp., and Pasteurella pneumotropica, were isolated from the eggshells. Other than Acinetobacter baumanni and Pasteurella pneumotropica most of the isolates were known to frequently appear during egg production processing.

• Parasites, Hosts and Diseases
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• v.55 no.6
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• pp.601-606
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• 2017

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.