• Title/Summary/Keyword: microbial assay

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Anti-microbial Activity of Saussurea lappa C.B. Clarke Roots

  • Chang, Kyung-Mi;Choi, Soo-Im;Chung, Sophia J.;Kim, Gun-Hee
    • Preventive Nutrition and Food Science
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    • v.16 no.4
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    • pp.376-380
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    • 2011
  • We investigated the total phenolic and flavonoid contents and the antimicrobial activity of ethanol extracts obtained from Saussurea lappa C.B. Clarke. The ethanol extracts of S. lappa C.B. Clarke were fractionated with various solvents (n-hexane, chloroform, and n-butanol). The antimicrobial activity of S. lappa C.B. Clarke was examined by disc-diffusion and micro-dilution susceptibility assays with six food-borne pathogens, and compared to that of the synthetic antibiotics. It is found that the S. lappa C.B. Clarke ethanol extract and n-hexane fraction have strong activity against B. cereus and V. parahaemolyticus strains compared to ampicillin. The inhibitory concentration ($IC_{50}$) values of hexane fraction against L. monocytogenes, B. cereus, and B. subtilis were 62.5, 250 and 500 ppm, respectively. Therefore, these data suggest that S. lappa C.B. Clarke may be useful as antimicrobial agents against food-borne pathogens.

Detection of Clostridium perfringens and its toxinotypes by enzyme linked immunosorbent assay from enterotoxaemic goats in Bangladesh

  • Islam, K.B.M.S.;Rahman, M.S.;Ershaduzzaman, Md.;Taimur, M.J.F.A.;Jang, Hyung-Kwan;Song, Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.33 no.1
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    • pp.37-44
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    • 2010
  • An enzyme-linked immnnosorbent assay (ELISA) has been performed for the detection of the prevailing toxinotypes of Clostridium perfringens obtained from conventional culturing of intestinal contents of goats which have died of suspected enterotoxaemia. The test was found effective to detect the toxins as well as types of the organism with less time and labor. The most prevailing type of C. perfringens causing enterotoxaemia in goat was C. perfringens type D (68.75%) and followed by C. perfringens type B (25%) and C (6.25%). No C. perfringens type A was detected. This study showed an intelligible picture of prevailing toxinotypes of C. perfringens in goats in Bangladesh. The use of the ELISA for the detection of clostridial types and toxins allows the differential diagnosis of C. perfringens types A, B, C and D enterotoxaemias from samples of intestinal contents and the typing of cultures of C. perfringens.

Plant Growth Promotion in Soil by Some Inoculated Microorganisms

  • Jeon, Jong-Soo;Lee, Sang-Soo;Kim, Hyoun-Young;Ahn, Tae-Seok;Song, Hong-Gyu
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.271-276
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    • 2003
  • The inoculation of some microorganisms into a microcosm containing soil from a barren lakeside area at Lake Paro in Kangwon-do enhanced plant growth significantly. The direct and viable counts of soil bacteria and soil microbial activities measured by electron transport system assay and fluorescein diacetate hydrolysis assay were higher in inoculated soil. The plant growth promoting effect of this inoculation may be caused by phytohormone production and the solubilization of insoluble phosphates by the inoculated bacteria. Three inoculated strains of Pseudomonas fluorescens produced several plant growth promoting phytohormones, including indole-3-acetic acid (auxin), which was confirmed by thin layer chromatography and GC/MS. P. fluorescens strain B16 and M45 produced 502.4 and 206.1 mg/l of soluble phosphate from Ca3(PO4)2 and hydroxyapatite, respectively. Bacillus megaterium showed similar solubilization rates of insoluble phosphates to those of Pseudomonas spp. We believe that this plant growth promoting capability may be used for the rapid revegetation of barren or disturbed land.

Features and Functions of Purple Pigment Compound in Halophytic Plant Suaeda japonica : Antioxidant/Anticancer Activities and Osmolyte Function in Halotolerance

  • Chung, Sang Ho
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.342-354
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    • 2018
  • Suaeda japonica is a halophytic plant that lives in mudflat at intertidal zone of western and southern coastal areas of Korea. The seawater-living plants showed a purple color during their whole life. In contrast, freshwater-living plants displayed a green color in leaves. When seawater-living plants were transferred to potting soil, the purple color was gradually changed to green in the leaves. The extracted purple pigment compound exhibited typical characteristics of betacyanin that were represented by water solubility, pH- and temperature-dependent color changes, sensitivity to light, UV-Vis spectra, and gel electrophoretic migration pattern. The LC-MS analysis of the extracted pigment compound showed the presence of two major protonated molecular ions ($[M+H]^+$) at m/z 651.1 and m/z 827.1. Antioxidant activity of the pigment compound was determined using stable free radical DPPH assay. It was found to have an antioxidant activity that is linearly increased in proportion to the reaction time for up to 30 min, and the activity was comparable to that of control BHA at 9.0 mg/ml. The anticancer activity against several tumor cell lines was also examined following the MTT assay. The significant growth inhibitory effect was observed on two tumor cell lines, SW-156 (human kidney carcinoma) and HEC-1B (human endometrial adenocarcinoma). Probably, the pigment compound may function as an osmolyte to uphold halotolerant physiological processes in saline environment.

The Antioxidant and Skin-whitening Effects of Saccharomyces cerevisiae FT4-4 Isolated from Berries Grown in Sunchang (화장품 소재로서 순창 베리류 유래 Sacchromyces cerevisiae FT4-4의 항산화 활성 및 미백 효과)

  • Seo, Ji won;Ryu, Myeong Seon;Yang, Hee-Jong;Jeong, Su-Ji;Jeong, Do-Youn
    • Journal of Life Science
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    • v.31 no.2
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    • pp.175-182
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    • 2021
  • Saccharomyces lysate has the well-known function of soothing the skin in various ways: it is an anti-irritant and can treat skin care conditions, such as skin whitening and antioxidative activity. However, data on the safety for use of Saccharomyces lysate in cosmetics and skin care products are still limited. To design a new cosmetic material with antioxidant and skin-whitening effects, 80 yeast strains were isolated from berries grown in Sunchang. Among the isolates, the FT4-4 strain, which exhibited superior biological activities, was selected for further experiments. The FT4-4 strain was identified as Saccharomyces cerevisiae by 18S rRNA gene sequencing analysis. S. cerevisiae FT4-4 showed higher DPPH radical-scavenging (51.41%), superoxide dismutase (62.23%), and tyrosinase inhibition (64.75%) activities. The highest yield of biomass (3.16 g/l) and maximum growth rate of S. cerevisiae FT4-4 were observed within 16 h. Furthermore, the cytotoxicity potential of S. cerevisiae FT4-4 on B16F10 melanoma cells was measured by an MTT assay, and the results indicated that S. cerevisiae FT4-4 had a capacity to inhibit melanin up to 72.02% at an initial 10 mg/ml concentration. These results suggest that S. cerevisiae FT4-4 could be a promising candidate as a multi-functional material for application in the cosmetic industry, especially because of its antioxidant and skin-whitening effects.

Effect of rearing system (free-range vs cage) on gut and muscle histomorphology and microbial loads of Italian White breed rabbits

  • Caterina Losacco;Antonella Tinelli;Angela Dambrosio;Nicoletta C. Quaglia;Letizia Passantino;Michele Schiavitto;Giuseppe Passantino;Vito Laudadio;Nicola Zizzo;Vincenzo Tufarelli
    • Animal Bioscience
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    • v.37 no.1
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    • pp.151-160
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    • 2024
  • Objective: The growing consumers' interest on animal welfare has raised the request of products obtained by alternative rearing systems. The present study was conducted to assess the influence of housing system on gut and muscle morphology and on microbial load in rabbits reared under free-range (FR) and cage system (CS). Methods: A total of forty weaned (35 days of age) male Italian White breed rabbits were allotted according to the rearing system, and at 91 days of age were randomly selected and slaughtered for the morphological evaluation of tissue from duodenum and longissimus lumborum. Morphometric analysis of the villus height, villus width, crypt depth, villus height/crypt depth ratio, and villus surface was performed. The microbial loads on hind muscle was determined by total mesophilic aerobic count (TMAC), Escherichia coli and Enterobacteriaceae; whereas, total anaerobic bacteria count (TABC) and TMAC, E. coli and Enterobacteriaceae was determined on caecal content. Results: Rearing system did not interfere with the duodenum and muscle histomorphology in both rabbit groups. Similarly, microbial load of caecal content showed no significant differences on the TABC and TMAC. Conversely, significant difference was found for E. coli strains in caecal content, with the lower counts in FR compared to CS rabbits (p<0.01). Microbiological assay of muscle revealed significant lower TMAC in FR vs CS rabbits (p< 0.05). All rabbit meat samples were negative for E. Coli and Enterobacteriaceae. Conclusion: Free-range could be considered a possible alternative and sustainable rearing system in rabbits to preserve gut environment and muscle quality.

Effects of Supplementation of Mixed Methanogens and Rumen Cellulolytic Bacteria on Biochemical Methane Potential with Pig Slurry (양돈슬러리를 이용한 혐기소화에서 미생물 첨가가 메탄발생에 미치는 영향)

  • Kim, Ji-Ae;Yoon, Young-Man;Jeong, Kwang-Hwa;Kim, Chang-Hyun
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.6
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    • pp.1049-1057
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    • 2012
  • The study investigated the biochemical methane potential (BMP) assay of pig slurry supplemented with mixed methanogens and cellulolytic bacteria to improve anaerobic digestion for methane production. For the BMP assay, 7 different microbial supplementation groups consisted of the cultures of mixed methanogens (M), Fibrobacter succinogenes (FS), Ruminococcus flavefaciensn (RF), R. albus (RA), RA+FS, M+RA+FS, and control. The cultures were added in the batch reactors with the increasing dose levels of 1% (0.5 mL), 3% (1.5 mL) and 5% (2.5 mL). Incubation for the BMP assay was carried out for 60 days at $38^{\circ}C$ using anaerobic digestate obtained from an anaerobic digester with pig slurry as inoculum. In results, 5% RF and RA+FS increased total biogas up to 8.1 and 8.4%, respectively, compared with that of control (p<0.05). All 5% microbial culture supplements significantly increased methane production up to 12.1~17.9% compared with that of control (p<0.05). Total solid (TS) and volatile solid (VS) digestion efficiencies showed no relationship to the increased supplementation levels of microbial cultures. After incubation, pH values in all treatment groups ranged between 7.527 and 7.657 indicating that methanogensis was not inhibited during the incubation. In conclusion, the results indicated that both hydrolysis and methanogenesis stages for methane production in anaerobic batch reactors were influenced by the supplemented microorganisms due to the chemical characteristics of pig slurry, but only the 5% supplementation level of all microbial culture supplements used in the experiment affected methane production.

H2AX Directly Interacts with BRCA1 and BARD1 via its NLS and BRCT Domain Respectively in vitro (H2AX의 BRCA1 NLS domain과 BARD1 BRCT domain 각각과의 in vitro 상호 결합)

  • Bae, Seung-Hee;Lee, Sun-Mi;Kim, Su-Mi;Choe, Tae-Boo;Kim, Cha-Soon;Seong, Ki-Moon;Jin, Young-Woo;An, Sung-Kwan
    • KSBB Journal
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    • v.24 no.4
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    • pp.403-409
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    • 2009
  • H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.

Antiproliferative and Cytotoxic Effects of Resveratrol in Mitochondria-Mediated Apoptosis in Rat B103 Neuroblastoma Cells

  • Rahman, Md. Ataur;Kim, Nam-Ho;Kim, Seung-Hyuk;Oh, Sung-Min;Huh, Sung-Oh
    • The Korean Journal of Physiology and Pharmacology
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    • v.16 no.5
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    • pp.321-326
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    • 2012
  • Resveratrol, a natural compound, has been shown to possess anti-cancer, anti-aging, anti-inflammatory, anti-microbial, and neuroprotective activities. In this study, we examined the antiproliferative and cytotoxicity properties of resveratrol in Rat B103 neuroblastoma cells; although it's molecular mechanisms for the biological effects are not fully defined. Here, we examined the cellular cytotoxicity of resveratrol by cell viability assay, antiproliferation by BrdU assay, DNA fragmentation by DNA ladder assay, activation of caspases and Bcl-2 family proteins were detected by western blot analyses. The results of our investigation suggest that resveratrol increased cellular cytotoxicity of Rat B103 neuroblastoma cells in a dose-and time-dependent manner with $IC_{50}$ of 17.86 ${\mu}M$ at 48 h. On the other hand, incubation of neuroblastoma cells with resveratrol resulted in S-phase cell cycle arrests which dose-dependently and significantly reduced BrdU positive cells through the downregulation of cyclin D1 protein. In addition, resveratrol dose-dependently and significantly downregulated the expression of anti-apoptotic protein includes Bcl-2, Bcl-xL and Mcl-1 and also activates cleavage caspase-9 and-3 via the downregulation of procaspase-9 and -3 in a dose-dependent manner which indicates that involvement of intrinsic mitochondria-mediated apoptotic pathway. In conclusion, resveratrol increases cellular cytotoxicity and inhibits the proliferation of B103 neuroblastoma cells by inducing mitochondria-mediated intrinsic caspase dependent pathway which suggests this natural compound could be used as therapeutic purposes for neuroblastoma malignancies.

The Cytotoxic and Apoptotic Effect of Pseudomonas aeruginosa Exotoxin A on Human Leukemia K-562 Cells (인간 백혈병 세포에서 Psuedomonas aeruginosa exotoxin A에 대한 세포독성과 세포자멸사 효과)

  • Chang, Jeong-Hyun;Kwon, Heun-Young
    • Korean Journal of Clinical Laboratory Science
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    • v.39 no.2
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    • pp.68-75
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    • 2007
  • After reports on regression of cancer in humans and animals infected with microbial pathogens date back more than 100 years, much effort has been spent over the years in developing a wild type or attenuated bacterial and purified bacterial proteins for the treatment of cancer. Pseudomonas aeruginosa exotoxin A (ETA) is known to inhibit cell growth and trigger significant cell death in various cancer cells. Although ETA induces apoptosis of cancer cells, its exact mechanism of action is not known yet. Four different assays were performed in this study: morphological assessment of apoptotic cells, cell cytotoxity, annexin-V binding assay, and cell cycle analysis. The proliferation and survival of the K-562 cells treated with ETA were decreased in a dose dependent manner. In addition, the apoptotic body of K-562 cells was induced by ETA treatment in a dose dependent manner. The ETA-induced apoptosis was confirmed by annexin-V binding assay. Flow cytometric analysis was examined to ascertain whether ETA could arrest the cell cycle at the sub-G1 phase. Our results suggest that P. aeruginosa ETA inhibits cell growth and induces apoptosis in human leukemia K-562 cells.

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