• Title/Summary/Keyword: Microbiological assay

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Isolation of High-molecular-weight-compound degrading microorganisms and sulfate reducing Bacteria involved in Composting Process (퇴비화 과정에 관여하는 생체 고분자 분해 미생물 및 황산 환원균의 분리)

  • Lee, Seong-Taek;Lee, Jae-Jeong;Na, Hyun-Jun
    • Journal of the Korea Organic Resources Recycling Association
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    • v.2 no.2
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    • pp.31-37
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    • 1994
  • For a microbiological study of composting process, screening and assay method for biopolymer degrading enzymes and microorganisms were developed and for the study of the possibility of composting in anaerobic state, distribution of sulfate reducing bacteria which plays a final role in anaerobic degradation was investigated. Substrates used for the development of assay methods for biopolymer degradation are ${\beta}-glucan$, xylan, dextran, CMC(carboxy methly cellulose), casein, and collagen. These substrates were made insoluble by a cross-linking agent and linked with dye to make chromogenic substrates. ${\beta}-glucan$ and xylan substrates could substitute congo-red method for screening of polymer degrading microorganisms without damaging the colonies. Sulfate reducing bacteria contained in the sample sludge showed preference to lactic acid, propionic acid, butyric acid and formic acid and could use acetic acid and valeric acid.

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Investigation of Waterborne Parasites in Drinking Water Sources of Ankara, Turkey

  • Bilal Bakir;Mehmet Tanyuksel;Fatma Saylam;Sultan Tanriverdi;R. engin Araz;Hacim, Ali-Kasim;Metin Hasde
    • Journal of Microbiology
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    • v.41 no.2
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    • pp.148-151
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    • 2003
  • Waterborne parasite infections are considered a reemerging threat. Most studies on the epidemiology of human cryptosporidiosis, giardiasis, and amebiasis have been carried out in developed countries, and there is little data on the occurrence of these infections in other areas. The objective of this study was to investigate the presence of waterborne parasites such as Cryptosporidium parvum, Giardia lamblia and Entamoeba histolytica in various water samples in Ankara, turkey. A total of 85 samples were examined, 43 from the municipal water supply, 34 from wells, 6 from the Ankara River, and 2 from two untreated dams; by conventional microscopy, immunologically and by polymerase chain reaction (PCR). Oocysts of C. parvum and cysts of G. lamblia were detected by using an indirect fluorescence (antigen) assay, whereas an enzyme linked immunosorbent assay was used to detect the cysts of E. histolytica and E. dispar. In addition, PCR was used for E. histolytica, E. dispar, C. parvum and G. lamblia detection. G. lamblia was found in 2 of the 34 well water samples, and parasites were found in 3 of the 6 Ankara River samples. The 1$\^$st/ contained E. histolytica cysts and Strongyloides stercoralis larvae. the 2$\^$nd/ E. histolytica cysts, and Trichuris trichiura eggs, and the 3$\^$rd/ C. parvum oocysts only. No parasite was observed in the municipal water samples and untreated dam water samples. These results extend our knowledge on waterborne parasites, such occurrence information on waterborne pathogens assists the management and treatment of municipal water.

Biodegradation of Medium-chain-length Polyhydroxyalkanoates by Pseudomonas sp. RY-1 (Pseudomonas sp. RY-1에 의한 Medium-chain-length Polyhydroxyalkanoates의 생분해)

  • 류강은;김영백;양영기;이영하
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.84-90
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    • 2000
  • Biodegradation of vanous medium-chain-length polyhydroxyalkanoates (MCL-PHAs) by an extracellular depolymerase system from Pseudomonas sp. RY-1 was investigated under laboratoly conditions. The degradation rate of the polymers was determined by quantitative clem zone technique, enzyme (turbidity) assay, and respirometry assay. Although the enzyme system secreted by Pscudomor~as sp. RY-1 was capable of degrading all MCL-PHAs tested. its secretion was influenced by the availability of secondary carbon sources. The rate of enzymatic degradation of MCL-PHAs was dependent upou the monomeric composition of the polyesters and reduced as the chain lengths of the monomer m t s in the polyesters increased. MCL-PHAs containing C-even monomer units showed faster degradation rate than MCL-PHAs containing C-odd monomer units. Respiration rates of MCL-PHAs with C-even monomer uuts were also much faster than those of MCL-PHAs with C-odd monomer units. The degmdation rate of MCL-PHAs bearing unsaturated substituents was faster than that of mcl-PHAs without functional substituents, which is suggesting the correlation between the degradation rate and the crystallinity of MCL-PHAs.

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Biochemical Studies on Korean Fermented Foods. VIII Studies on Vitamin $B_{12}$ Contents of the Fermented Foods in Korea (한국 발효식품에 대한 생물화학적 연구 ( 제 8 보) 발효식품중의 Vitamin $B_{12}$의 함량 조사보고)

  • Lee, In-Jae;Haw, Kum;Kim, Sung-Ikk
    • YAKHAK HOEJI
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    • v.4 no.1
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    • pp.50-52
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    • 1959
  • In order to know contents of vitamin B$_{12}$ in the fermented foods in Korea, the contents of this vitamin are studied on the following foods; kimchies (pickled vegetables) .................. 39 chukkals (fermented and salted sea foods) ...... 8 fermented soy-bean products .................... 5 the kimchies and fermented soybean products studied in this paper, are from individual homes and chukkals are from markets. The content of vitamin B$_{12}$ is estimated by the microbiological assay method using lactobacillus leichmannii ATCC 7830. Details for assay are indicated in the original part of this paper. The pseudo-vitamin B$_{12}$ substances as thymidine which is active to the growth of lactobacillus leichmannii, are eliminated by the alkali treatment method of sample solution. According to the results as indicated in the table, the following conclusions are summerized; 1. Vitamin B$_{12}$ contents of kimchies for winter season are 1.03 to 1.52 mcg% in average. The hobakk-kimchi which contain the highest content of vitamin B$_{12}$ is not popular one among Korean. 2. Chukkals contain much higher content of vitamin B$_{12}$ varying from 0.91 to 11.10mcg%. 3. The soybean fermented foods, as kanjang, denjang, kochojang, contain lower content of vitamine B$_{12}$ varying from 0.08 to 0.52 mcg% containing higher content of pseudo-vitamin. 4. Based on daily consumption of kimchi in winter season by Chai, the consumption of this vitamin through kimchi is about 3 mcgs daily per capita. This will be a quite source of this vitamin for Korean as vitamin C. as vitamin C.

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Biochemical Studies on Korean Fermented Foods. (IX) Variation of Vitamin $B_{12}$ during the Kimchi Fermentation Period (한국 발효식품에 대한 생물화학적 연구 (제9보) 침채류의 발효에 따르는 $VitaminB_{12}$의 변화에 대하여)

  • Lee, In-Jae;Haw, Kum;Kim, Sung-Ikk
    • YAKHAK HOEJI
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    • v.4 no.1
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    • pp.53-55
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    • 1959
  • In order to confirm on production of vitamin $B_{12}$ during the kimchi fermentation period, the variation of its content is studied in this paper As a sample of kimchi for this fermentation study due to the seasonal condition, nabakkimchi which is aseasonal one in early spring, is prepared by author. The content of vitamin $B_{12}$ is estimated by the microbiological assay method using lactobacillus leichmannii A Tee 7830. Details for assay are indicated in the original part. And the pseudo-vitamin $B_{12}$ substances as thymidine which is active to the growth of lactobacillus leichmannii, are eliminated by the alkali treatment method of sample solution, According to the results shown in table 2 and 3 and figures 1 to 4, the following conclusions are summerized; 1. Vitamin $B_{12}$ produced during the fermentation period of kimchi. And the content of this vitamin during its optimum fermentation period, is much higher content compared with the theoretically calculated amount from its materials, This has been confirmed at the repeated experiment. The trend of variation during its period is indicated at the figures. 2. The sterilized kimchi inhibited the fermentation by sterilization at the preparation, did not increased its content showing only its theoretically calculated amount.

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Antibody Response to Crude Cell Lysate of Propionibacterium acnes and Induction of Pro-Inflammatory Cytokines in Patients with Acne and Normal Healthy Subjects

  • Basal, E.;Jain, A.;Kaushal, G.P.
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.117-125
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    • 2004
  • Propionibacterium acnes (P. acnes) plays an important role in the disease pathogenesis of acne vulgaris, a disorder of pilosebaceous follicles, seen primarily in the adolescent age group. In the present study, the presence of antibodies against P. acnes (MTCC1951) were detected in acne patient (n=50) and disease free controls (n=25) using dot-ELISA and Western blot assay. The ability of P. acnes to induce pro-inflammatory cytokines by human peripheral blood mononuclear cells (PBMCs), obtained from acne patients and healthy subjects, were also analysed. The patients (n=26) who were culture positive for skin swab culture, were found to have a more advanced disease and higher antibody titres (1:4000 to >1:16000) compared to the P. acnes negative patients (n=24) and normal controls (n=25). An analysis of patients' sera by western blot assay recognized a number of antigenic components of P. acnes, rang-ing from 29 to 205 kDa. The major reactive component was an approximately 96 kDa polypeptide, which was recognised in 92% (24 of 26) of the patients sera. Further, the P. acnes culture supernatant, crude cell lysate and heat killed P. acnes whole cells, obtained from 72-h incubation culture, were observed to be able to induce significant amounts of IL-8 and tumor necrosis factor alpha (TNF-${\alpha}$) by the PBMCs in both the healthy subjects and patients, as analysed by cytokine-ELISA. The levels of cytokines were significantly higher in the patients than the healthy subjects. A major 96 kDa polypep-tide reactant was eluted from the gel and was found to cause dose dependent stimulation of the pro-ductions of IL-8 and TNF-${\alpha}$. Thus, the above results suggest that both humoral and pro-inflammatory responses play major roles in the pathogenesis of acne.

Development of Human Papillomavirus DNA Array by Using Lateral Flow Membrane Assay (Lateral Flow Membrane를 이용한 인유두종 바이러스 DNA Array의 개발)

  • Kim, Ki-Whang;Lee, Hyung-Ku;Cho, Hong-Bum
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.346-351
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    • 2008
  • This study develops DNA array which can detect specific sequence of human papilomavirus (HPV) by using lateral flow membrane assay which is usually used for point of care test including pregnant diagnosis. Principle of HPV DNA array is as follow; fixing DNA probe which is peculiar to HPV type 6, 11, 16, 18, 31, 45 on a surface of lateral flow membrane and inducing hybridization response between probe and HPV PCR products which is obtained by using biotin-labeled MY09/l1 primers. And then, we can see the result of DNA hybridization that streptavidin labelled colloidal gold is responded with hybrid biotin. Lateral flow membrane array developed in this study confirms major HPV type economically and conveniently compared with existing HPV DNA chip method.

Cultural Characteristics of a Recombinant Saccharomyces cerevisiae for the Improved Production of a Antibacterial Peptide Defensin of Fleshfly (쉬파리 유래 항균텝티드 Defensin의 생산 증진을 위한 재조합 Saccharomyces cerevisiae의 배양학적 특성)

  • 안종석;강대욱;이준원;김민수;김보연;오원근;민태익
    • Korean Journal of Microbiology
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    • v.36 no.3
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    • pp.236-241
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    • 2000
  • A defensin is an inducible antibacterial peptide from a fleshfly and contains 40 residues basic peptide with six cysteines. For the consiruction of recombinant S cerevisiae expressing defensin, the structural gene coding for active defensin was chemically synthesized and fused in fiam to GAP promoter, MFul preprosequence and the GAL7 transcription terminator, generating a recombinant plasnlid pGMD18. S. ce~evisine 2805 Gells were transror~ned to uracil prototroph by the pGMDl8 arid the transformed cells showing antibacterial activity against 111. luteus TAM1056 were selected by growth inhibition zone assay. The optimal culture conditions for the unprovement of the defensin production of a selected tmdonnant were investigated. The optirmzed medium containing 0.4% yeast extract, 2% corn steep liquor, 2.5% glucose and 0.05% $C_2CO_3$, could be determined and the optimum lemperature. and initial pH could be detennnied as $28^{\circ}C$ and pH 3, ~mpectively. The optimized conditioiis revealed the trvofold Increase in the cell growth and the fourfold in the antibaclerial activity. coinpar-ed with tllc Yl'D medium.

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Antimicrobial activity of fraction mixture of ethanol extracts from Eucalyptus globulus, Yucca recurvifolia, and Melaleuca alternifolia against several human skin microbes (유칼립투스, 유카와 차나무의 추출분획 혼합물의 여러 인간 피부 상재균에 대한 항균활성)

  • Lee, Da-Sol;Hong, In Kee;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.46-51
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    • 2019
  • This study was carried out to evaluate antimicrobial effects of a mixture of resin fractionated ethanol extract of Eucalyptus globulus, Yucca recurvifolia, and tea tree (Melaleuca alternifolia). The plant fraction mixture showed low minimum inhibitory concentration (0.24~3.32 mg/ml) against several bacteria and yeast that usually used as the target skin microbes in a cosmetic industry, and it was more effective than antibiotics, triclosan and ampicillin. In a time-kill assay the plant fraction mixture reduced more than 92% of microbial populations during 4 h, and significantly increased leakage of nucleotides from all microorganisms tested. Antimicrobial effect of the plant fraction mixture was not affected by divalent cation ($Mg^{2+}$ and $Ca^{2+}$). These results suggest that the fraction mixture of ethanol extracts of E. globulus, Y. recurvifolia, and M. alternifolia may be utilized as an efficient preservative in cosmetics to prevent contamination by human skin microbes.

Yeast two-hybrid assay with fluorescence reporter (형광 리포터를 활용한 효모 단백질 잡종 기법 개발)

  • Park, Seong Kyun;Seo, Su Ryeon;Hwang, Byung Joon
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.199-205
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    • 2019
  • Yeast two-hybrid (Y2H) technique has been used to study protein-protein interactions, but its application particularly to a large-scale analysis of protein interaction networks, is limited by the fact that the technique is labor-intensive, based on scoring colonies on plate. Here, we develop a new reporter for the measurement of the protein-protein interactions by flow cytometry. The yeast harboring interacting proteins can also be enriched by fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS). When two interacting proteins are present in the same yeast cell, a reporter protein containing 10 tandem repeats of c-myc epitope becomes localized on the surface of the cell wall, without affecting cell growth. We successful measured the surface display of c-myc epitope upon interacting p53 with SV40 T antigen by flow cytometry. Thus, the newly developed Y2H assay based on the display of c-myc repeat on yeast cell wall could be used to the simultaneous analysis of multiple protein-protein interactions without laborious counting colonies on plate.