• Title/Summary/Keyword: inoculum

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Inhibitory Effect of Metal Surface on the Antimicrobial Resistance Microorganism (금속표면이 항생제 내성균주의 생육억제에 미치는 영향)

  • Kim, Jung-Beom;Kim, Jae-Kwang;Kim, Hyunjung;Cho, Eun Jung;Park, Yeon-Joon;Lee, Hae Kyung
    • Annals of Clinical Microbiology
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    • v.21 no.4
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    • pp.80-85
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    • 2018
  • Background: The aim of this study was to comparatively evaluate the bactericidal effects of copper, brass (copper 78%, tin 22%), and stainless steel against methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VREFM), and multidrug-resistant Pseudomonas aeruginosa (MRPA). Methods: The isolates (MRSA, VREFM, MRPA) used in this study were mixed wild type 3 strains isolated from patients treated at Uijeongbu St. Mary's Hospital in 2017. These strains showed patterns of multidrug resistance. The lyophilized strains were inoculated into and incubated for 24 hr in tryptic soy broth at $35^{\circ}C$. The initial bacterial inoculum concentration was adjusted to $10^5CFU/mL$. A 100-mL bacterial suspension was incubated in containers made of brass (copper 78%, tin 22%), copper (above 99% purity), and stainless steel at $35^{\circ}C$. Viable counts of bacteria strains were measured for 9 days. Results: In this study, the bactericidal effects of copper and brass on MRSA, VREFM, and MRPA were verified. The bactericidal effect of stainless steel was much weaker than those of copper and brass. The bactericidal effect was stronger on MRPA than on MRSA or VREFM. Conclusion: To prevent cross infection of multidrug resistant bacteria in hospitals, further studies of longer duration are needed for testing of copper materials on objects such as door knobs, faucets, and bed rails.

Effect of Physical Control Technology on Aspergillus ochraceus Reduction (물리적 제어기술이 Aspergillus ochraceus 저감화에 미치는 영향)

  • Lee, Eun-Seon;Kim, Jong-Hui;Kim, Bu-Min;Oh, Mi-Hwa
    • Journal of Food Hygiene and Safety
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    • v.36 no.5
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    • pp.447-453
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    • 2021
  • In this study, the effectiveness of physical control technology, a combined light sterilization (LED, UV) and hot water treatment in reducing Aspergillus ochraceus for food production environment was investigated. In brief, 1 mL aliquot of A. ochraceus spore suspension (107-8 spore/mL) was inoculated onto stainless steel chips, which was then dried at 37℃, and each was subjected to different physical treatment. Treatments were performed for 0.5, 1, 2, 5, 8, and 11 hours to reduce the strains using a light-emitting diode, but no significant difference was confirmed among the treatments. However, a significant reduction was observed on the chips treated with UV-C exposure and hot water immersion. After being treated solely with 360 kJ/m2 of UV-C on stainless steel chip, the fungi were significantly reduced to 1.27 log CFU/cm2. Concerning the hot water treatment, the initial inoculum amount of 6.49 log CFU/cm2 was entirely killed by immersion in 83℃ water for 5 minutes. Maintaining a high temperature for 5 minutes at the site is difficult. Thus, considering economic feasibility and usability, we attempted to confirm the appropriate A. ochraceus reduction conditions by combining a relatively low temperature of 60℃ and UV rays. With the combined treatments, even in lukewarm water, A. ochraceus decreased significantly through the increases in the immersion time and the amount of UV-C irradiation, and the yield was below the detection limit. Based on these results, if work tools are immersed in 60℃ lukewarm water for 3 minutes and then placed in a UV sterilization device for more than 10 minutes, the possibility of A. ochraceus cross-contamination during work is expected to be reduced.

Molecular Breeding of Tobacco Plants Resistant to TMV and PVY (분자생물학적 TMV 및 PVY 저항성 연초 육종)

  • E.K. Pank;Kim, Y.H.;Kim, S.S.;Park, S.W.;Lee, C.H.;K.H.Paik
    • Proceedings of the Korean Society of Tobacco Science Conference
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    • 1997.10a
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    • pp.134-152
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    • 1997
  • Plant viruses of tobacco including tobacco mosaic virus (TMV) and potato virus Y (PVY) cause severe economic losses in leaf-tobacco production. Cultural practices do not provide sufficient control against the viruses. Use of valuable resistant cultivars is most recommendable for the control of the viruses. However, conventional breeding programs are not always proper for the development of virus-resistant plants mostly owing to the frequent lack of genetic sources and introduction of their unwanted properties. Therefore, we tried to develop virus-resistant tobacco plants by transforming commercial tobacco cultivars, NC 82 and Burley 21, with coat protein (CP) or replicase (Nlb) genes of TMV and PVY necrosis strain (PVY-VN) with or without untranslated region (UTR) and with or without mutation. Each cDNA was cloned and inserted in plant expression vectors with 1 or 2 CaMV 35S promotors, and introduced into tobacco leaf tissues by Agrobacterium tumefaciens LBA 4404. Plants were regenerated in kanamycin-containing MS media. Regenerated plants were tested for resistance to TMV and PVY In these studies, we could obtain a TMV-resistant transgenic line transformed with TMV CP and 6 genetic lines with PVY-VN cDNAs out of 8 CP and replicase genes. In this presentation, resistance rates, verification of gene introduction in resistant plants, stability of resistance through generations, characteristics of viral multiplication and translocation in resistant plants, and resistance responses relative to inoculum potential and to various PVY strains will be shown. Yield and quality of leaf tobacco of a promising resistant tobacco line will be presented.

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Analysis of the Reduction Effect of Combined Treatment with UV-C and Organic Acid to Reduce Aspergillus ochraceus and Rhodotorula mucilaginosa Contamination (Aspergillus ochraceus와 Rhodotorula mucilaginosa 저감을 위한 자외선과 유기산 복합처리 효과 분석)

  • Eun-Seon Lee;Jong-Hui Kim;Bu-Min Kim;Mi-Hwa Oh
    • Journal of Food Hygiene and Safety
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    • v.39 no.1
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    • pp.54-60
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    • 2024
  • This study investigated the effectiveness of using pathogens and aqueous acids to reduce the Aspergillus ochraceus and Rhodotorula mucilaginosa contamination in livestock production environments. For this study, 1 mL of each bacterial suspension (107-108 spores/mL) was inoculated on a knife surface, dried at 37℃, and used under each treatment condition. First, to investigate the effect of organic acids, acetic, lactic, and citric acids were used. Subsequently, to select the appropriate concentration, they were prepared at concentrations of 0.5, 1, 2, 3, 4, and 5%, respectively. Accordingly, to further maximize the effect of organic acid treatment, we combined the treatment with ultraviolet light. The two strains showed a significant difference (P<0.05) compared to the initial strain, with a greater than 90% decrease in the concentrations of all organic acids. Consequently, acetic and lactic acids decreased by approximately 5 and 2 log colony forming unit (CFU)/cm2, respectively, when treated with ultraviolet light (360 mJ/cm2); however, citric acid decreased by less than 1 log CFU/cm2. However, when manufactured with 4% acetic acid, a severe malodor was emitted, making it difficult for workers to use it in a production environment. Accordingly, the optimal treatment conditions for organic acid and ultraviolet light for application were selected as follows: immersion in a 4% lactic acid solution for 1 minute and then, sterilization with ultraviolet light at 360 mJ/cm2. Finally, when a pork meat sample was cut with a knife that was finally washed with lactic acid and treated with ultraviolet light, the low level of inoculum transferred from the cleaned knife to the surface of the sample was not detected. In conclusion, using this established method can prevent cross-contamination of the surface of the meat during processing.

Epidemiology and Control of Rice Blast in Korea (한국(韓國)에서의 도열병(病) 발생(發生), 만연(蔓延)과 그 방제(防除))

  • Park, Jong Seong
    • Korean Journal of Agricultural Science
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    • v.12 no.2
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    • pp.356-369
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    • 1985
  • In Korea, inevitable researches for the blast control exactly started from 1927 by the organization of Office of Rural Development with the local extensive outbreak of panicle blast at Jeonlla Buk-Do Province in 1926. At present, the rice blast is still one of the most destructive and widespread diseases in spite of considerable contributions by rice scientists, particularly plant pathologists during last 55 years in Korea. Rice blast control and management are very difficult because of the marked variability in pathogenicity of the blast fungus. From the results obtained through the disease surveys during last 70 years, different 3 prevalence type of blast such as bimodal leaf-blast type, bimodal panicle-blast type and bimodal continual blast type were recognized. In generally speaking, pattern of blast outbreak is said to be characterized by severe outbreak of panicle blast after slight outbreak of leaf blast with discontinuity between leaf and panicle blast. So we have to pay much attention for successful management of panicle blast giving direct influence to rice yield. Main factors induce blast epidemic were pointed out to be breakdown of the disease resistance, nutritional unbalance such as excess application of nitrogen, delay of transplantation and longspell of rain fall by extensive surveys and researches on blast during last 70 years in Korea. The fact some of Japonica varieties such as Kokuryomiyako, Tamanishiki, Ginbozu and Pungok belong to varietal group A had been cultivated with extensive acrage over 30 years in this country should be mentioned by Korean rice scientists. Differences in field resistance between varieties in the same group are detectable and apparently small but sometimes epidemiologically significant differential effects may be found out in case of blast. Much more attention should be payed to accumulate the knowledges on field resistance for successful management of blast. Excess application of nitrogen is more effective to outbreak of panicle blast than that of leaf blast of IR varieties. In comparatively low level application of nitrogen infection rate of panicle blast of IR varieties is considerably high. Low temperature effects on outbreak of blast is very great. It results in remarkable increase of the inoculum potential on the leaf lesions and infection of panicle blast in leaf sheathes of IR varieties during the booting stage. In economic point of view, it is concluded that 5 times sprays of effective fungicides including 3 times before and 2 times after heading is good enough to control blast. We have experienced no one of control measures for blast is superior to all others. The integrated control measures was established as guideline of blast control around 1950 in Korea. This guideline must be helpful for rice growers as long as rice growing continue.

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Studies on the Citric Acid Fermentation with Fungi (Part III) Citric Acid Fermentation with Selected Strains (사상균에 의한 구연산발효에 관한 연구 (제III보) 선정균에 의한 구연산발효)

  • 성낙계;김명찬;심기환;정덕화
    • Microbiology and Biotechnology Letters
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    • v.8 no.3
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    • pp.181-191
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    • 1980
  • For the purpose of studies on the citric acid production, some experiments were carried out with isolated strains. The results obtained were as follows. 1) The optimal culture media of the strain M-80 in surface culture contained 140g of sucrose, 3.0g of (N $H_4$)$_2$S $O_4$, 1.5g of K $H_2$P $O_4$, 0.2g of MgS $O_4$.7$H_2O$, 3.0mg of F $e^{++}$, 1.0mg of Z $n^{++}$, 0.5N HCI to a pH of 5.0 and distilled water to 1.0 liter; and that of the strain M-315 in surface culture contained 140g of sucrose, 2.0g of N $H_4$N $O_3$, 1.0g of K $H_2$P $O_4$, 0.25g of MgS $O_4$. 7$H_2O$, 2.0mg of F $e^{++}$, 2.0mg of Z $n^{++}$, 0.05mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. While that of the strain M-315 in submerged culture contained 140g of sucrose, 2.5g of N $H_4$N $O_3$, 1.5g of K $H_2$P $O_4$, 0.3g of MgS $O_4$. 7$H_2O$, 3.0mg of F $e^{++}$, 0.1mg of C $u^{++}$, 0.5N HCI to a pH of 4.5 and distilled water to 1.0 liter. The optimal temperature and size of inoculum were mostly 28-3$0^{\circ}C$, 10$^{7}$ -10$^{8}$ spores/50ml, respectively. 2) Through the course of citric acid production, the growth of strains had nearly been completed, pH value was rapidly decreased below 2.0 and the content of sugar was also reduced, while the accumulation of citric acid in media was remarkably begun in about 3-4 days. The yields of citric acid generally reached the maximum level in 8-10 days in surface or submerged fermentation process. 3) Methanol was effective citric acid production when they were added to fermentation media. In the case of surface culture, by addition of 2% (strain M-80), 3% (strain M-315), the yields of citric acid was increased 6.5%, 20.6%, respectively and 5.0% yield was increased by addition of 3% methanol in submerged culture media of the strain M-315. 4) Chromatography analysis of culture broth after fermentation under optimal culture conditions detected that the majority of acid in media was citric acid. 72.1mg/ml, 98.1mg/ml, of citric acid were determined in surface culture media by strains of M-80, M-315, and 59.8 mg/ml of citric acid was contained in the submerged culture media by the strain M-315. strain M-315.

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Measuring Intracellular Mycobacterial Killing Using a Human Whole Blood Assay (인체 전혈 모델을 이용한 세포내 결핵균 살균력에 관한 연구)

  • Cheon, Seon-Hee;Song, Ho-Yeon;Lee, Eun-Hee;Oh, Hee-Jung;Kang, In-Sook;Cho, Ji-Yoon;Hong, Young-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.497-509
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    • 2002
  • Background : The mechanisms through which cellular activation results in intracellular mycobacterial killing is only partially understood. However, in vitro studies of human immunity to Mycobacterium tuberculosis have been largely modeled on the work reported by Crowle, which is complicated by several factors. The whole blood culture is simple and allows the simultaneous analysis of the relationship between bacterial killing and the effect of effector cells and humoral factors. In this study, we attempted to determine the extent to which M. tuberculosis is killed in a human whole blood culture and to explore the role of the host and microbial factor in this process. Methods : The PPD positive subject were compared to the umbilical cord blood and patients with tuberculosis, diabetes and lung cancer. The culture is performed using heparinized whole blood diluted with a culture medium and infected with a low number of M. avium or M. tuberculosis $H_{37}Ra$ for 4 days by rotating the culture in a $37^{\circ}C$, 5% $CO_2$ incubator. In some experiments, methlprednisolone- or pentoxifyline were used to inhibit the immune response. To assess the role of the T-cell subsets, CD4+, CD8+ T-cells or both were removed from the blood using magnetic beads. The ${\Delta}$ log killing ratio was defined using a CFU assay as the difference in the log number of viable organisms in the completed culture compared to the inoculum. Results : 1. A trend was noted toward the improved killing of mycobacteria in PPD+ subjects comparing to the umbilical cord blood but there was no specific difference in the patients with tuberculosis, diabetes and lung cancer. 2. Methylprednisolone and pentoxifyline adversely affected the killing in the PPD+ subjects umbilical cord blood and patients with tuberculosis. 3. The deletion of CD4+ or CD8+ T-lymphocytes adversely affected the killing of M. avium and M. tuberculosis $H_{37}Ra$ by PPD+ subjects. Deletion of both cell types had an additive effect, particularly in M. tuberculosis $H_{37}Ra$. 4. A significantly improved mycobacterial killing was noted after chemotherapy in patients with tuberculosis and the ${\Delta}$ logKR continuously decreased in a 3 and 4 days of whole blood culture. Conclusion : The in vitro bactericidal assay by human whole blood culture model was settled using a CFU assay. However, the host immunity to M. tuberculosis was not apparent in the human whole blood culture bactericidal assay, and patients with tuberculosis showed markedly improved bacterial killing after anti-tuberculous chemotherapy compared to before. The simplicity of a whole blood culture facilitates its inclusion in a clinical trial and it may have a potential role as a surrogate marker in a TB vaccine trial.

Effect of VA Mycorrhizal Fungi on Alleviation of Salt Injury in Hot Pepper (Capsicum annuum L.) (VA 균근균(菌根菌) 접종(接種)에 의한 고추의 염류장해(鹽類障害) 경감효과(輕減效果))

  • Sohn, Bo-Kyoon;Huh, Sang-Man;Kim, Kil-Yong;Kim, Yong-Woong
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.6
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    • pp.482-492
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    • 2000
  • Vesicular arbuscular mycorrhizal (VAM) fungi are known to increase plant growth as well as to enhance salt tolerance of plants where plant roots are colonized by VAM. In pot experiment, pepper was grown in soil containing 0, 200, 400, and $600P\;kg\;ha^{-1}$ with and without mycorrhizal inoculum. Pots were irrigated with saline water containing 0.5, 2.0, and $6.0dS\;m^{-1}$. At 0, 200, and $400P\;kg\;ha^{-1}$ of three EC treatments, plant hight in mycorrhizal treatments was significantly different compared to nonmycorrhizal treatments. However, plant hight at $600P\;kg\;ha^{-1}$ was not different between mycorrhizal and nomycorrhizal treatments. Leaf area at $0P\;kg\;ha^{-1}$ of three EC treatments, and $200P\;kg\;ha^{-1}$ of $6.0dS\;m^{-1}$ in mycorrhizal treatments significantly increased compared to nonmycorrhizal treatments. However, these increase were not discovered in high salinity and P level. Level of EC affected dry weight, and especially, interection of P and EC, or P and VA inoculation highly affected root dry weight. R/S ratio generally decreased in mycorrhizal treatments. Significantly decreased R/S ratio was shown at 0, 400, and $600P\;kg\;ha^{-1}$ of $6.0dS\;m^{-1}$. Chlorophyll content generally increased with decreased salinity and P level where mycorrhizal treatments showed higher chlorophyll content compared to nonmycorrhizal treatments. The benefits of VAM inoculation on fruit production was discovered at only low P level and salinity. Mycorrhizal dependency on dry weight basis was generally shown in $0P\;kg\;ha^{-1}$ of three EC treatments and 0.5, $2.0dS\;m^{-1}$ of $200P\;kg\;ha^{-1}$ level. Colonization rate ranged 3.3 to 43.3% and number of spores was 47.7 to 198.3 $100g^{-1}$ soil. Colonization rate and number of spores increased with decreased P level and salinity where there was high correlation ($r=0.858^{**}$) between both. Also improved uptake of mineral nutrients was discovered at mycorrhizal treatments in decreased P level and salinity.

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CELL CULTURE STUDIES OF MAREK'S DISEASE ETIOLOGICAL AGENT (조직배양(組織培養)에 의한 Marek 병(病) 병원체(病原體)의 연구(硏究))

  • Kim, Uh-Ho
    • Korean Journal of Veterinary Research
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    • v.9 no.1
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    • pp.23-62
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    • 1969
  • Throughout the studies the following experimental results were obtained and are summarized: 1. Multiplication of agents in primary cell cultures of both GF classical and CR-64 acute strain of Marek's disease infected chicken kidneys was accompanied by the formation of distinct transformed cell foci. This characteristic nature of cell transformation was passaged regularly by addition of dispersed cell from infected cultures to normal chicken kidney cell cultures, and also transferred was the nature of cell transformation to normal chick-embryo liver and neuroglial cell cultures. No cytopathic changes were noticed in inoculated chick-embryo fibroblast cultures. 2. The same cytopathic effects were noticed in normal kidney cell monolayers after the inoculation of whole blood and huffy coat cells derived from both forms of Marek's disease infected chickens. In these cases, however, the number of transformed cell foci appearing was far less than that of uninoculated monolayers prepared directly from the kidneys of Marek's disease infected chickens. 3. The change in cell culture IS regarded as a specific cell transformation focus induced by an oncogenic virus rather than it plaque in slowly progressing cytopathic effect by non-oncogenic viruses, and it is quite similar to RSV focus in chick-embryo fibroblasts in many respects. 4. The infective agent (cell transformable) were extremely cell-associated and could not be separated in an infective state from cells under the experimental conditions. 5. The focus assay of these agents was valid as shown by the high degree of linear correlation (r=0.97 and 0.99) between the relative infected cell concentration (in inoculum) and the transformed cell foci counted. 6. No differences were observed between the GF classical strain and the CR-64 acute strain of Marek's disease as far as cell culture behavior. 7. Characterization of the isolates by physical and chemical treatments, development of internuclear inclusions in Infected cells, and nucleic acid typing by differential stainings and cytochemical treatments indicated that the natures of these cell transformation agents closely resemble to those described fer the group B herpes viruses. 8. Susceptible chicks inoculated with infected kidney tissue culture cells developed specific lesions of Marek's disease, and in a case of prolonged observation after inoculation (5 weeks) the birds developed clinical symptoms and gross lesions of Marek's disease. Kidney cell cultures prepared from those inoculated birds and sacrificed showed a superior recovery of cell transformation property by formation of distinct foci. 9. Electron microscopic study of infected kidney culture cells (GF agent) by negative staining technique revealed virus particles furnishing the properties of herpes viruses. The particle was measured about $100m{\mu}$ and, so far, no herpes virus envelop has been seen from these preparations. 10. No relationship of both isolates to avian leukosis/sarcoma group viruses and PPLO was observed.

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