• Title/Summary/Keyword: colony forming unit

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Detection of Aspergillus and Penicillium genera by Enzyme-Linked Immunosorbent Assay Using a Monoclonal Antibody

  • Kwak, Bo-Yeon;Shon, Dong-Hwa;Kwon, Byung-Joon;Kweon, Chang-Hee;Lee, Ke-Ho
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.21-28
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    • 2001
  • Enzyme linked-immunosorbent assay (ELISA) for a rapid detection of fungi, Aspergillus and Penicillium genera in food, were developed and their efficiencies were approved by detecting artificially contaminated agricultural commodities. Mice were immunized with partially purified Aspergillus flavus extracellualr polysaccharide (EPS) and lymph node cells of the mice were fused with the myeloma cells for production of monoclonal antibodies. Mab 1G11, one of the antibodies, was selected and purified. A sandwich ELISA was established and its detection limit toward A. flavus EPS was 1mg/ml. Among the 59 strains tested (including 18 species of Aspergillus, 16 of Penicillium, 11 of Fusarium, 1 of Absidia, 2 of Alternaria, 2 of Candida, 2 of Cladosporium, 2 of Geotrichum, 2 of Mucor, 2 of Rhizopus, 1 of Trichoderma), species of Aspergillus and penicillium had a high reactivity with Mab 1G11 even up to 10,000 times dilution of culture broths. The other genera except Cladosporium resinae showed no reactivity, thus Mab 1G11 was specific to the genera of Aspergillus and Penicillium. The epitope of A. flavus EPS against monoclonal Mab 1G11 was on the carbohydrate moiety when 1 to 100$\mu g/g$ A. flavus EPS were put into rice, potato, and mandarin orange, the average recoveries detected by sandwich ELIA were 123, 59, and 76%, respectively. Correlation was found to be linear between the EPS, and mycelium of A. flavus and Penicillium citrinum grown in a liquid medium (r=0.87 and 0.96), and also between the EPS and colony forming unit in solid media of rice of potato (r=0.91-0.99).

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Increase in Aminotransferase Levels during Urinary Tract Infections in Children

  • Park, Ju Yi;Ko, Kyung Ok;Lim, Jae Woo;Cheon, Eun Jeong;Yoon, Jung Min
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.16 no.2
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    • pp.89-94
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    • 2013
  • Purpose: The aim of this study was to evaluate the prevalence of increased aminotransferase levels and to identify associated factors in children admitted to hospital with urinary tract infections (UTIs). Methods: The study included children with a diagnosis of UTI who were admitted to the Konyang University Hospital from January 2007 to May 2011. The total number of patients was 249 and the mean age was $15.88{\pm}28.21$ months. UTI was defined as a positive urine culture (> $10^5$/colony forming unit [CFU]) with pyrexia. Patients were treated by intravenous antibiotics, such as ampicillin/sulbactam, aminoglycoside, cephalosporins or vancomycin. Patients with neonatal jaundice or other liver disease were excluded. We investigated the relationship of aminotransferase levels with the type of antibiotic, degree of vesicoureteral reflux (VUR), and causative organisms. Results: Children with increased aminotransferase levels were younger than those with normal levels (p=0.001), but white blood cell count, platelet count, causative organisms, type of antibiotics and presence of VUR were not associated with aminotransferase levels. Aminotransferase levels became normal within 1 month after discharge without special measures, except in 1 case. Conclusion: We found that many children with UTI have abnormal aminotransferase levels. In most cases, this change is mild and self-limiting. We conclude that increased aminotransferase level increase during UTI do not require unnecessary tests and excessive treatment.

Radioprotective effect of fucoidan against hematopoietic and small intestinal stem cells of γ-ray irradiated mice (감마선을 조사한 마우스의 조혈 및 소장줄기세포에 대한 fucoidan의 방호효과)

  • Park, Eunjin;Jeon, Seong Mo;Joo, Hong-Gu;Hwang, Kyu-Kye;Jee, Youngheun
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.393-399
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    • 2008
  • We investigated the potential of fucoidan for its ability to provide protection from gamma rayinduced damage. In our results, the fucoidan significantly improved the counts of endogenous colony forming unit to $9.5 {\pm} 1.5$, from $5.5 {\pm} 2.5$ compared with un-treated irradiated control group at 10 day after 7 Gy whole body irradiation. After 2 Gy irradiation, fucoidan treatment attenuated the percent of tail DNA of splenocytes, parameters of DNA damage, from $30.17 {\pm} 1.7%$ to $13.67 {\pm} 2.81%$ 2.81% by comet assay and also accelerated the proliferation of splenocytes, compared with un-treated irradiated control group by 3Hthymidine incorporation assay. Furthermore, fucoidan decreased the number of apoptotic fragments per intestinal crypt by 31.8% at 1 days after 2 Gy irradiation. These results indicated that the fucoidan significantly improved the hematopoietic recovery, prevented the DNA damage in immune cells and enhanced their proliferation, which had been suppressed by ionizing radiation. in addition, fucoidan rescued intestinal cells from radiation-induced apoptosis. Thus, this study raises the possibility of using fucoidan as adjuvant therapeutic agent after radiotherapy.

Lactobacillus casei strain C1 attenuates vascular changes in spontaneously hypertensive rats

  • Yap, Wei Boon;Ahmad, Faisal Malau;Lim, Yi Cheng;Zainalabidin, Satirah
    • The Korean Journal of Physiology and Pharmacology
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    • v.20 no.6
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    • pp.621-628
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    • 2016
  • Hypertension can be caused by various factors while the predominant causes include increase in body fluid volume and resistance in the circulatory system that elevate the blood pressure. Consumption of probiotics has been proven to attenuate hypertension; however, the effect is much strain-dependent. In this study, a newly isolated Lactobacillus casei (Lb. casei ) strain C1 was investigated for its antihypertensive properties in spontaneously hypertensive rats (SHR). Lactic acid bacteria (LAB) suspension of 11 log colony-forming unit (CFU) was given to SHR (SHR+LAB, n=8), and phosphate buffer saline (PBS) was given as a control in SHR (SHR, n=8) and in Wistar rats as sham (WIS, n=8). The treatment was given via oral gavage for 8 weeks. The results showed that the weekly systolic blood pressure (SBP), mean arterial pressure (MAP), diastolic blood pressure (DBP) and aortic reactivity function were remarkably improved after 8 weeks of bacterial administration in SHR+LAB. These effects were mostly attributed by restoration of wall tension and tensile stress following the bacterial treatment. Although not statistically significant, the level of malondialdehye (MDA) in SHR+LAB serum was found declining. Increased levels of glutathione (GSH) and nitric oxide (NO) in SHR+LAB serum suggested that the bacterium exerted vascular protection through antioxidative functions and relatively high NO level that induced vasodilation. Collectively, Lb. casei strain C1 is a promising alternative for hypertension improvement.

Characteristics and response of mouse bone marrow derived novel low adherent mesenchymal stem cells acquired by quantification of extracellular matrix

  • Zheng, Ri-Cheng;Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Joo-Hee;Park, Ji-Man
    • The Journal of Advanced Prosthodontics
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    • v.6 no.5
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    • pp.351-360
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    • 2014
  • PURPOSE. The aim of present study was to identify characteristic and response of mouse bone marrow (BM) derived low-adherent bone marrow mesenchymal stem cells (BMMSCs) obtained by quantification of extracellular matrix (ECM). MATERIALS AND METHODS. Non-adherent cells acquired by ECM coated dishes were termed low-adherent BMMSCs and these cells were analyzed by in vitro and in vivo methods, including colony forming unit fibroblast (CFU-f), bromodeoxyuridine (BrdU), multi-potential differentiation, flow cytometry and transplantation into nude mouse to measure the bone formation ability of these low-adherent BMMSCs. Titanium (Ti) discs with machined and anodized surfaces were prepared. Adherent and low-adherent BMMSCs were cultured on the Ti discs for testing their proliferation. RESULTS. The amount of CFU-f cells was significantly higher when non-adherent cells were cultured on ECM coated dishes, which was made by 7 days culturing of adherent BMMSCs. Low-adherent BMMSCs had proliferation and differentiation potential as adherent BMMSCs in vitro. The mean amount bone formation of adherent and low-adherent BMMSCs was also investigated in vivo. There was higher cell proliferation appearance in adherent and low-adherent BMMSCs seeded on anodized Ti discs than machined Ti discs by time. CONCLUSION. Low-adherent BMMSCs acquired by ECM from non-adherent cell populations maintained potential characteristic similar to those of the adherent BMMSCs and therefore could be used effectively as adherent BMMSCs in clinic.

Effects of Temperature and Moisture on the Survival of Colletotrichum acutatum, the Causal Agent of Pepper Anthracnose in Soil and Pepper Fruit Debris

  • Kang, Beum-Kwan;Kim, Joo-Hyeong;Lee, Kyeong-Hee;Lim, Sang-Cheol;Ji, Jae-Jun;Lee, Jong-Won;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.25 no.2
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    • pp.128-135
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    • 2009
  • The survival of Colletotrichum acutatum was investigated in soil, infected fruits, and infected fruit debris incorporated into soil at several temperatures with different soil moisture levels. Samples were examined at 2-week intervals for 18 weeks to determine the survival of the pathogen based on the number of colony forming unit (CFU) of C. acutatum recovered on a semi-selective medium. C. acutatum conidia survived in both sterile and non-sterile soil at 4 and $10^{\circ}C$ for 18 weeks. If infected pepper fruits were completely dried, C. acutatum survived for 18 weeks at temperature from 4 to $20^{\circ}C$. Soil temperature and moisture affected the survival of C. acutatum in infected fruit debris incorporated into soil after air-drying. The effect of soil moisture on survival was weaker at low temperatures than at high temperatures. For up to 16 weeks, conidia were recovered from fruit debris in soil that had been kept at 4 to $20^{\circ}C$ and below 6% soil moisture. Conidia were recovered from fields until approximately 6 months after pepper fruits were harvested. Using PCR with species-specific primers and a pathogenicity test, we identified conidia recovered from soil and infected fruit from both the laboratory and field as C. acutatum and as the primary inoculum causing pepper anthracnose.

Fluorogenic and Chromogenic Assay for Rapid Detection of Escherichia coli and Total Coliform Bacteria (효소발색법을 이용한 대장균 및 총대장균군 신속 검사)

  • Lee, Keun-Heon;Kim, Hun-Soo;Kim, Byong-Ryol;Lee, Seung-Hui;In, Chi-Kyung;Park, Kyeong-Ryang
    • Journal of Korean Society on Water Environment
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    • v.25 no.3
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    • pp.363-369
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    • 2009
  • We developed the Eco medium for Escherichia coli and total coliforms, which was modified by Violet Red Bile (VRB) medium, and derived the standard curve of exponential phase at $OD_{410}$ by using type strains such as E. coli ATCC11303, Enterobacter cloacae KCTC2361, Klebsiella pneumoniae KCTC2241, and Citrobacter freundii KCTC2359. Also, we used total 93 samples of spring and stream water to compare the detection ability of total coliforms between the method using Eco medium and such as most probable number (MPN), and plate count methods. As a result, the qualitative analysis of E. coli and total coliforms using Eco medium contained ortho-nitrophenyl-$\beta$-galactoside (ONPG) and 4-methylumbelliferyl-$\beta$-D-glucuronide (MUG) was same as those of Korean standard methods (Colilert kit). And the colony forming unit (CFU) detected in Eco medium was similar to those of result from MPN and plate count methods. Moreover, the agreement, sensitivity, and specificity of the developed kit was more than 97.5% in comparison with Colilert kit for 350 samples. Thus, the Eco medium can be used both qualitative and quantitative analysis of E. coli and total coliforms.

Expression of the E. coli LacZ Gene in Chicken Embryos Using Replication Defective Retroviral Vectors Packaged With Vesicular Stomatitis Virus G Glycoprotein Envelopes

  • Kim, Teoan;Lee, Young Man;Lee, Hoon Taek;Heo, Young Tae;Yom, Heng-Cherl;Kwon, Mo Sun;Koo, Bon Chul;Whang, Key;Roh, Kwang Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.2
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    • pp.163-169
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    • 2001
  • Despite the high potency of the retrovirus vector system in gene transfer, one of the main drawbacks of has been difficulty in preparing highly concentrated virus stock. Numerous efforts to boost the virus titer have ended in unsatisfactory results mainly due to fragile property of retrovirus envelope protein. In this study, to overcome this problem, we constructed our own retrovirus vector system producing vector viruses encapsulated with VSV-G (vesicular stomatitis virus G glycoprotein). Concentration process of the virus stock by ultracentrifuge did not sacrifice the virus infectivity, resulting in more than 108 to 109 CFU (colony forming unit) per ml on most of the target cell lines tested. Application of this high-titer retrovirus vector system was tested on chicken embryos. Injection of virus stock beneath the blastoderms of pre-incubated fertilized eggs resulted in chick embryos expressing E. coli LacZ gene with 100% efficiency. Therefore, our results suggest that it is possible to transfer the foreign gene into chicken embryo using our high-titer retrovirus vector.

Bacteriological Hazard Analysis in Minimally Processed Shellfish Products Purchased from Korean Seafood Retail Outlets (유통 중인 패류 단순가공품의 세균학적 위해요소 분석)

  • Kim, Hyun-Jung;Lee, Dong-Soo;Lee, Ji-Min;Kim, Young-Mog;Shin, Il-Shik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.2
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    • pp.121-126
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    • 2018
  • The purpose of this study was to assess bacteriological hazards in the following 7 kinds of minimally processed shellfish products purchased from Korean seafood retail outlets: raw oysters stored with seawater in polyethylene bags, frozen oysters, raw mussels, frozen mussels, boiled mussels, raw short-necked clam, and frozen short-necked clams, obtained from Korean seafood retail outlets. The total coliform and Escherichia coli counts determined in all samples were detected below regulation limit of the Korean government guidelines (Food Code). In addition, the high-risk bacterial pathogens, Listeria monocytogenes, Salmonella spp., and E. coli O157:H7 were not detected in any samples. Low-risk pathogens such as Staphylococcus aureus and Vibrio parahaemolyticus were detected at levels above $1.0{\times}10^2$ colony forming unit (CFU)/g in some minimally processed shellfish products. Notably, S. aureus was detected in all samples. Raw oysters stored with seawater in polyethylene bags, frozen oysters, and boiled blue mussels are commonly ingested without heating, and therefore these minimally processed shellfish products pose bacteriological hazards. The detection of S. aureus in all shellfish products, an organism indicative of poor personal hygiene, which can grow and multiply during distribution, indicates the necessity of improving the food safety of minimally processed shellfish products.

An Experimental Study on Artificial Infection of Olive Flounder Paralichthys olivaceus by Streptococcus parauberis Using Different Injection Sites (넙치(Paralichthys olivaceus)의 Streptococcus parauberis 인위감염을 위한 공격실험 방법에 관한 연구)

  • Kim, Tae-Ho;Lee, Nam-Sil;Choi, Hye-Sung;Jung, Sung-Hee;Han, Hyun-Ja
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.53 no.4
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    • pp.628-636
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    • 2020
  • Streptococcosis in the olive flounder Paralichthys olivaceus can be caused by Streptococcus parauberis. We compared three routes of administration for experimental injections of the S. parauberis 19FBSPa0003 strain in the olive flounder. Pathological changes were observed during the experimental infection. Inflammation of the serous membrane in the liver, intestine, spleen and heart was the major pathological change found in the infected olive flounder. No mortality was observed in fish that received intraperitoneal (IP) injection at less than 1×104 colony-forming unit (CFU)/fish. The lethal dose 50 for olive flounder, given an intravenous (IV) injection, was 7.94×104 CFU/fish. Fish with a higher concentration of IV injected S. parauberis (1×108 CFU/fish) died within a maximum of two days. However, serious necrosis and bacterial proliferation in ellipsoidal cells of the spleen and heart tissues were found in moribund or dead fish, 1-2 days after IV injection. Similar histopathological signs were observed in olive flounder inoculated by subcutaneous (SC) infected and naturally infected. In addition, SC was also strongly associated with bacteria concentration and cumulative mortality rate. Based on these results, SC is the recommended method for artificial infection by S. parauberis in the olive flounder.