• Title/Summary/Keyword: time isolates

Search Result 335, Processing Time 0.028 seconds

Molecular Cloning and Expression of Heat-stable Enterotoxin Gene from Swine Enterotoxigenic Escherichia coli (돼지에서 유래한 병원성 대장균의 내열성 장독소 생산유전자의 Cloning 및 발현)

  • 김교창;도대흥
    • Journal of Food Hygiene and Safety
    • /
    • v.6 no.3
    • /
    • pp.147-155
    • /
    • 1991
  • Enterotoxigenic E. coli is one of the major causative agents of the infantile diarrhea and traveler's diarrhea. The heat-stable enterotoxin(ST) is thought to be a virulence factor in the pathogenesis of the diarrhea and to be a maker for identification of the enterotoxingeic E. coli from non pathogenic E. coli. The isolate of enterotoxigenlc E. coli was isolated from swine during 1989 year(from 5 to 10 month) in the Kyong-gi and Chung-Cheong provinces, and three strains(KM-4, KM-7 and KM-12) was selected from 189 isolates of ST producing E. coli. The detection of a ST produced of the isolated E. coli was performed by the infant mouse assay(IMA). This study was designed to know optimal conditions for the production of the ST and the molecular properties of plasmids of the enterotoxigenic E. coli. Amount of ST produced were the most at initial pH 8.5~9.0 of succinate salts medium culture. The cultural time of the same medium was accumulated the highest level of ST was at the 14 to 16 hours, and then stationary phase was at the 20 hours. From this experiment the KM-7 strain was selected among ST producing strains by IMA. Partial plasmid-curing experiment was done to select plasmid encoding for ST among other plasmids and then comparing the plasmid pattern of ST producing strain(KM-7) with those of other ST non-producing strains, it is found that ST gene exists on the about 80 Kbp plasmid. Each fragment of this plasmid digested with EcoRl was ligated to vector pBR 322 and transformed into E. coli K-12. A clone producing ST(eKT 53) was selected by IMA. The EcoRl digestion pattern of the isolated plasmid(pKD 37) from the ST producing clone it is indicated that the size of the inserted fragment in eKT 53 strain is 16 Kbp. The cultured supernatant of eKT 53 strain was positive result of ST production in IMA.

  • PDF

Antibacterial activity of lactic acid bacteria isolated from traditional fermented foods and development of a starter for fermented milk (전통발효식품에서 분리한 유산균의 항균 활성 및 발효유 스타터 개발)

  • Park, Jong-Hyuk;Moon, Hye-Jung;Oh, Jeon-Hui;Lee, Joo-Hee;Choi, Kyung-Min;Cha, Jeong-Dan;Lee, Tae-Bum;Lee, Min-Jeong;Jung, Hoo-Kil
    • Food Science and Preservation
    • /
    • v.20 no.5
    • /
    • pp.712-719
    • /
    • 2013
  • This study was conducted to investigate the antibacterial activity of lactic acid bacteria isolated from traditional fermented foods and to develop a new starter for fermented milk. The isolates were identified using 16S rDNA sequencing and named Lactobacillus plantarum A, Leuconostoc lactis B and L. acidophilus C. The activity of these strains to inhibit the growth of food-borne human pathogens (Escherichia coli NCTC 12923, Salmonella Typhimurium NCTC 12023, Listeria monocytogenes NCTC 11994) was measured using the paper disc method. All these strains showed strong antibacterial activity against Li. monocytogenes NCTC 11994. The experiment groups were the fermented milks with these strains, and the control group was the fermented milk with the commercial starter (ABT 5). The change of pH, acidity and viable cell counts were measured during their aging time. All the experiment groups showed a significant difference in their aging times compared to the control group. However, the sensory test showed that the experiment groups can be used as useful starters for fermented milk. This result suggests that L. plantarum A, Leu. lactis B and L. acidophilus C have the potential to be developed as new starters for fermented milk.

Epidemiological characteristics of 2002 outbreak of classical swine fever in Korea (2002년 한국에서 발생한 돼지콜레라의 역학적 특성)

  • Park, Choi-Kyu;Song, Jae-Young;Wee, Sung-Hwan;Lee, Eune-Sub;Yoon, Hachung;Moon, Oun-Kyeong;Choi, Eun-Jin;Nam, Hyang-Mi
    • Korean Journal of Veterinary Research
    • /
    • v.46 no.2
    • /
    • pp.107-117
    • /
    • 2006
  • This paper described the epidemiological characteristics of 2002 outbreak of classical swine fever (CSF) in Korea. A total of thirteen CSF-infected farms could be classified into two clusters according to the location and time of outbreak. Two farms located in the same county of Gangwon province and 11 farms located in several different districts of Incheon metropolitan/Gyeonggi province were identified as CSF-infected from April 16 to 30 and from October 7 to December 21 in 2002, respectively. As the result of epidemiological analysis, the two clusters of outbreaks were turned out to be independent epidemics which had different sources of virus introduction. Three farms were found to have been infected primarily; one located in Cheolwon county of Gangwon province and two located in Kangwha county of Incheon metropolitan area. The most likely factors of virus introduction into these primary infected farms were considered to be direct or indirect contact by foreign workers and/or owners of the infected farms who had come back from traveling in China before outbreaks. This was supported by the genetic typing of CSF viruses isolated from the pigs of infected farms. All the virus isolates of 2002 outbreak were found to be genetic type 2, whereas the viruses isolated before 2000 were type 3 and the reference strains, such as attenuated live vaccine virus (LOM strain) and high virulent challenge virus (ALD strain), were type 1. Accordingly, we concluded that the 2002 CSF outbreak must have been caused by a newly introduced virus from overseas and the type 3 virus must have been eradicated after the last outbreak of 1999 by the national CSF eradication campaign which were implemented since 1996. Based on the combined analysis of epidemiological data and genetic typing, the transmission routes of classical swine fever virus were found to be the movement of vehicles (60%) and persons (10%), neighbourhood spread (20%) and unknown (10%). It is expected that the analyzed data and findings of classical swine fever outbreak epidemic could be very useful to establish the disease control and eradication program for the country in the future.

Synergistic Effects of Cinnamomum camphora Leaves Extract against Clinical Isolated Methicillin-resistant Staphylococcus aureus (임상에서 분리한 메치실린 내성 황색포도상구균에 대한 녹나무잎 추출물의 항균활성)

  • Cha, Jeong-Dan;Choi, Mi-Rae;Ko, Eun-Sil;Hwang, Seung-Mi;Kang, Jea-Ran;Oh, Joa-Sub;Park, Young-Jin;Jung, Young-Hwan;Jeon, Ah-Lim;Choi, Kyung-Min
    • Korean Journal of Plant Resources
    • /
    • v.28 no.1
    • /
    • pp.9-15
    • /
    • 2015
  • Methicillin-resistant Staphylococcus aureus (MRSA) has been emerging worldwide as one of the most important hospital and community pathogens. At the same time, because of the difficulty in developing chemical synthetic drugs and because of their side-effects, scientists are making more efforts to search for new drugs from plant resources to combat clinical multidrug-resistant microbial infections. Cinnamomum camphora (C. camphora) is a plant of family Lauraceae, and grown Jeju island in South Korea that are used as a drug to treat neurasthenia, epilepsy, cystitis, pyelonephritis, digitalis, cancer, and diabetes mellitus in folk remedies. In this study, antibacterial activites of 80% ethanol extract of C. camphora leaves (CCE) were investigated in combination with antibiotics against clinical isolates of MRSA. The results showed that CCE was determined with MIC and MBC values ranging from 156 to 313 and 313 to $625{\mu}g/ml$, oxacillin from 128 to 256 and 128 to $512{\mu}g/ml$, ampicillin from 4 to 64 and 8 to $128{\mu}g/ml$. The combination of CCE with oxacillin or/and ampicillin were synergistic effect against MRSA 1, 6, 7, 8, 10, 11, 12, 13, and 15/ MRSA 1, 2, 6, and 7.

Studies on the Enzymatic Partial Hydrolysis of Soybean Protein Isolates (효소처리에 의한 분리대두 단백질의 부분 가수분해에 관한 연구)

  • Lee, Cherl-Ho;Kim, Chan-Shick;Lee, Sam-Pin
    • Korean Journal of Food Science and Technology
    • /
    • v.16 no.2
    • /
    • pp.228-234
    • /
    • 1984
  • A partial hydrolysis of soybean protein isolate was carried out by using pepsin and trypsin. The degree of hydrolysis was evaluated by chemical analysis, viscometric measurements and gel electrophoresis. The functional properties of the hydrolyzates such as flow behavior, emulsion properties and foaming properties were evaluated. A selective hydrolysis of 11S protein fraction by pepsin was observed from the SDS-PAG electrophoresis. The changes in the molecular weight distribution by different conditions of enzyme hydrolysis were evaluated. The changes in the intrinsic viscosity of the protein hydrolylate by reaction time were highly correlated to the contents of TCA soluble protein and 0.03 M $CaCl_2$ soluble nitrogen. The degree of hydrolysis ($DH_{TCA}$, $DH_{Ca}$) were used to evaluate the effect of enzyme treatment on the functional properties of the hydrolyzate. The apparent viscosity and emulsion capacity and stability of the protein solution decreased as DH increased, while the foaming capacity increased linearly with the increasing DH.

  • PDF

Regulation of stf Operon Expression and Its Virulence (살모넬라가 발현하는 stf 오페론의 조절과 병원성 인자로서의 기능)

  • Kim Sam-Woong;Kim Young-Hee;Kang Ho-Young
    • Journal of Life Science
    • /
    • v.15 no.4 s.71
    • /
    • pp.553-560
    • /
    • 2005
  • The stf (Salmonella typhimurium fimbriae) operon consisting of stfA(CDEFG assumes to encode putative fimbriae. The complete stf operon is existed in S. typhimurium and S. choleraesuis, whereas it is absent in S. typhi. Analyses of the amino acid residues between major subunit StfA of the Stf fimbriae and those of known other fimbriaes suggested that Stf belongs to class I type fimbriae. Through comparison of StfD chaperone with the other fimbrial chaperones, and of C-terminus in subunits of Stf fimbriae, it belongs to FGS (with a short Fl-G1 loop) subfamily. In order to investigate the expression of stf operon, we have constructed a Salmonella strain containing a chromosomal stfA::lacZYA transcriptional fusion, resulting in S. typhimurium $_X8532$. The strain $_X8532$ lacked the expression of \beta-galactosidase$ under normal culture conditions. However, with longer incubation time of the S. typhimurium $_X8532$, we have isolated 21 individual strains exhibiting $Lac^+$ phenotype. $Lac^+$ phenotype was appeared as approximately 0.03 frequency per generation. All isolates expressed lacZ constitutively in the various environmental conditions. Various global regulatory proteins including RpoS, OmpR, and CpxR were not involved in the regulation of the stf operon. A S. typhimurium $_X8661$ mutant lacking stfAC function attenuated 6.7 folds more than that of wild type $_X3761$ in the mouse virulence test, suggesting in the somehow involved in the Salmonella pathogenesis.

Subspecies Classifying and Characterizing the Two Groups of Antagonistic Sorangium cellulosum against Botrytis cinerea and Colletotrichum acutatum (Botrytis cinerea와 Colletotrichum acutatum에 항균활성을 갖는 점액세균 Sorangium cellulosum에 대한 아종 분류 및 길항 특성 연구)

  • Koo, Tae-Hoon;Yun, Sung-Chul
    • Research in Plant Disease
    • /
    • v.24 no.3
    • /
    • pp.213-220
    • /
    • 2018
  • We classified the previously reported antagonistic strains of Sorangium cellulosum into 5 subspecies (A-E). Four strains were antagonistic to Botrytis cinerea (AB group) and two strains were antagonistic to Colletotrichum acutatum (AC group). According to the genetic and sequential analyses with standard genes, xynB1, bglA2, groEL1 for grouping, all strains of AB group were belonged to subspecies C and all strains of AC group were belonged to subspecies D. In addition, high pressure liquid chromatography with the culture filtrates confirmed the genetic results, because AB group had peaks with retention time at 20-22.5 minutes, whereas AC group had no peak. There was positive relationship ($R^2=0.9652$) between the control values of infecting B. cinerea on cherry tomatoes and the main peak areas of chromatograms among the four isolates of AB group. From the subspecies results of AB group, the main peak of KYC 3270 was expected to be epothilone D. However the retention times of the standard of commercial epothilone D and the main peak of KYC 3270 culture filtrate were different as 9.9 and 11.581 min., respectively. Finally, the antagonistic metabolite of AB group was inferred as 7-ketone epothilone D.

Anti-inflammatory Effects of the Extracts of Prunus pendula for. ascendens (Makino) Ohwi Leaves and Identification of Active Constituents (올벚나무 잎 추출물의 항염 효능 및 유효성분 규명)

  • Hong, Hye Jin;Ko, Ha Na;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.45 no.2
    • /
    • pp.117-129
    • /
    • 2019
  • In this study Anti-inflammatory activity was investigated for the extract of Prunus pendula for. ascendens (Makino) Ohwi by monitoring nitric oxide (NO) production in LPS-stimulated RAW 264.7 murine macrophage cells. The P. pendula ethyl acetate (EtOAc) fraction showed to decrease the NO synthesis by 76.3% at $100{\mu}g/mL$ concentration. The inhibition occurred in a dose-dependent manner without causing cell toxicity. The EtOAc fraction also inhibited the production of $PGE_2$, $IL-1{\beta}$, IL-6 and expression of iNOS, COX-2 protein in dose-dependent manner. From the phytochemical study to isolate the active constituents, five known compounds were identified, which are ursolic acid (1), prunasin (2), methyl p-coumarate (3), kaempferol (4), astragalin (5). All of the compounds 1 - 5 were isolated for the first time from the P. pendula. Among the isolates, the flavonoids 4 and 5 were verified to inhibit NO production with high efficiency. These results suggested that extract of P. pendula leaves could be useful as anti-inflammatory agents in pharmaceutical or cosmetic applications.

Growth of Candida albicans Biofilm is Inhibited by Salvia miltiorrhiza (단삼에 의한 Candida albicans 바이오필름 발달의 억제)

  • Lee, Heung-Shick;Kim, Younhee
    • Microbiology and Biotechnology Letters
    • /
    • v.47 no.3
    • /
    • pp.465-472
    • /
    • 2019
  • Candida albicans is an opportunistic human pathogen that causes infections. Candidiasis is often related to antifungal resistance because the pathogen has the ability to form biofilms. In a previous study, we found that the Salvia miltiorriza ethanol extract demonstrated anticandidal activity by altering membrane permeability and inhibiting the cell wall synthesis in C. albicans. Our results here demonstrate that $78{\mu}g/ml$ of the S. miltiorriza extract significantly diminished the early stage biofilms formed by 10 clinical C. albicans isolates by 51.3%; this was analyzed by 2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) reduction assay. The effect of the S. miltiorrhiza extract on the adhesion of C. albicans cells to polystyrene plates and germ tube formation was examined via microscopic investigation. Although the density of the adhered cells was remarkably reduced up on incubation with $39{\mu}g/ml$ S. miltiorrhiza extract, germ tube formation by C. albicans was rarely affected. Quantitative real-time PCR analysis showed that the S. miltiorrhiza extract downregulated the expression of C. albicans hypha-specific genes, EAP1 by 34.7% (p < 0.001), ALS1 by 45.0% (p < 0.001), ALS3 by 48.1% (p < 0.001), and ECE1 by 21.3% (p = 0.006), respectively. Our data suggest that the S. miltiorrhiza ethanol extract significantly inhibited the early stage of biofilm formation by C. albicans by interfering with cell adhesion, by downregulating EAP1, ALS1 and ALS3, and presumably by modifying the cell wall and membrane structure.

Anti-inflammatory and Anti-oxidative Constituents from the Extract of Cinnamomum yabunikkei Leaves (생달나무 잎 추출물 유래 항염 및 항산화 활성 성분)

  • Kim, So Hee;Kim, Jung Eun;Lee, Nam Ho
    • Journal of the Korean Chemical Society
    • /
    • v.65 no.1
    • /
    • pp.15-24
    • /
    • 2021
  • In this study, the extract of Cinnamomum yabunikkei leaves were investigated for the anti-inflammatory and anti-oxidative activities and their active constituents were identified. In the anti-inflammatory tests using LPS-stimulated RAW 264.7 cells, the ethyl acetate (EtOAc) fraction inhibited the production of nitric oxide (NO) without causing cell toxicity. In addition, the EtOAc fraction reduced expression of iNOS protein and production of pro-inflammatory cytokines (TNF-α, IL-1β). Upon the anti-oxidative studies by DPPH and ABTS+ radicals, potent radical scavenging activities were observed in the EtOAc fraction. Five phytochemicals were isolated from the extract of C. yabunikkei leaves; (4S,5R)-4-hydroxy-5-isopropyl-2-methylcyclohex-2-enone (1), methoxy-(3,5-dimethoxy-4-hydroxyphenyl)ethanediol (2), afzelin (3), nicotiflorine (4) and narcissin (5). As far as we know, compounds 1-5 were isolated for the first time from this plant. In the anti-inflammatory tests for the isolates, compound 1, 3, 4 and 5 were determined to decrease NO production without causing cell toxicity. Furthermore, compound 1 reduced expression of iNOS protein and exhibited potent inhibitory activities of pro-inflammatory cytokines (TNF-α, IL-1β, IL-6). Based on these results, it was suggested that the extract and isolated compounds from C. yabunikkei leaves could be potentially applicable as natural source for pharmaceutical and/or cosmetic ingredients.