• Title/Summary/Keyword: Agar diffusion

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Isolation of Salmonella from the layer chickens reacting in pullorum-typhoid agglutination test (추백리 혈청검사 양성 산란계로부터 Salmonella속균 분리)

  • 류재윤;전무형;장경수;손현수;곽학구;박경재;우용구
    • Korean Journal of Veterinary Service
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    • v.22 no.3
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    • pp.221-237
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    • 1999
  • To investigate the specificity of rapid slide agglutination test for pullorum-gallinarum diseases and to obtain a basic data for avian salmonellosis control, salmonella isolation was peformed for the layer chickens positively reacted in pullonlm-typhoid agglutination test. The biochemical, serological and antimicrobial properties of the isolates were examined. The results obtained through this study were summarized as follows; 1. Of 2,384 chickens tested by the agglutination test, 606 chickens (25.4%) were positive reactors. 154 of 606 reactors and 49 of the non-reacting chickens were investigated for salmonella isolation, resulting in isolation of 68 strains of salmonellae from 27 chickens. 2. By organs, the isolation frequency from liver, cecum, spleen, ovary and gall bladder showed 8.9% (18 strains), 8.9% (18 strains), 7.4% (15 strains), 4.4% (9 strains) and 3.9% (8 strains), respectively. 3. By culture medium the combination of selenite broth and MacConkey agar revealed the highest isolation rate and the enrichment culture by delayed secondary enrichment culture method was found the most effective for salmonella isolation. 4. The serotypes of 68 salmonella isolates were identified as 3 strains of S pullorum, 24 strains of S gallinarum, 15 strains of S typhimurium, 8 strains of S enteritidis, 7 strains of S paratyphi A, 5 strains of S typhimurium and 6 strains of the other salmonellae. 5. The serotypes of 8 salmonella strains isolated from 49 chickens non-reacting in pullorum-typhoid agglutination test were identified as 3 strains of S typhimurium and 5 strains of S infantis. 6. When 24 chickens of which 68 strains of salmonellae isolated were examined by microplate agglutination test, the average antibody titer for pullorum antigen was $2^{5.25}$. The chickens at antibody titer between $2^3$ and $2^5$ showed the higher frequency of isolation as compared with the chickens at the other titers. 7. When salmonella isolates were tested the antimicrobial drug sensitivity by disk diffusion method, S paratyphi A were highly sensitive by 100% to ATM and GM, S typhimurium, by 88% to AM, CIP, IMP and TN, S infantis, by 100% to AM, CRO, ENR and PIP, S enteritidis,by 100% to IMP and PIP, S pullorum, by 100% to ATM, CRO, ENR and PIP and S gallinarum, by 92% to CRO, CIP and PIP.

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Investigation on Antibacterial and Antioxidant Activities, Phenolic and Flavonoid Contents of Some Thai Edible Plants as an Alternative for Antibiotics

  • Lee, J.H.;Cho, S.;Paik, H.D.;Choi, C.W.;Nam, K.T.;Hwang, S.G.;Kim, Soo-Ki
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1461-1468
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    • 2014
  • This study was aimed to examine the antibacterial and antioxidative properties of seven edible plants from Thailand to develop alternative antibiotics as feed additives. The plants include Citrus aurantifolia Swingle (Lime) fruits and its leaves, Sesbania grandiflora L. (Agati sesbania) leaves, Piper sarmentosum Roxb (Wild betal) leaves, Curcuma domestica Valeton (Turmeric) roots, Morinda citrifolia L. (Beach mulberry) leaves, Cassia siamea britt (Siamea cassia) leaves, and Cocos nucifera L. (Coconut) peels. The plants were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. Antibacterial activities with minimum inhibitory concentration (MIC) were determined by agar diffusion assay against Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, and Clostridium perfringens that were considered pathogenic strains in livestock infection. Methanol extracts of C. aurantifolia Swingle fruits and leaves showed the broadest spectrum of antibacterial activities except for C. perfringens. Butanol extract of S. grandiflora L. leaves showed the strongest activity against Burkholderia sp. with MIC, $135{\mu}g/mL$. P. sarmentosum Roxb leaves showed antibacterial activities against E. coli, Burkholderia sp. and H. parasuis. Ethyl acetate and water extracts from C. domesitca Valeton roots showed MIC of $306{\mu}g/mL$ and $183{\mu}g/mL$, respectively against only C. perfringens. Antioxidative activity was determined by 2-diphenyl-2-picryl hydrazyl photometric assay. The methanol extracts of C. aurantifolia Swingle fruits and P. sarmentosum Roxb leaves showed the highest antioxidant activity among all the extracts with 3.46 mg/mL and 2.70 mg/mL effective concentration 50% ($EC_{50}$) values, respectively. Total contents of phenolics and flavonoids were measured from the plant extracts. Methanol extracts of S. grandiflora L. and chloroform extracts of C. domestica Valeton were found to have the highest amount of total phenolics, 41.7 and $47.8{\mu}g/mL$, respectively. Flavonoid content of methanol extracts in S. grandiflora L. T was $22.5{\mu}g/mL$ and the highest among plant extracts tested. These results indicated that C. aurantifolia Swingle, S. grandiflora L., P. sarmentosum Roxb, and C. domestica Valeton have antibacterial and antioxidant activities and can be used as alternative antibiotics or potential feed additives for the control of animal pathogenic bacteria.

Selection of Lactic Acid Bacteria with Antibacterial Activity for Extension of Kimchi Shelf-life (김치의 저장성 향상을 위한 항균활성 우수 유산균 선발)

  • Choi, Hak-Jong;Kim, Yu Jin;Lee, Na Ra;Park, Hae Woong;Jang, Ja Young;Park, Sung-Hee;Kang, Miran;Kim, Hyun Ju;Lee, Jong-Hee;Lee, Jong-Hoon;Pyun, Yu-Ryang;Kim, Tae-Woon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.328-332
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    • 2014
  • A survey was conducted on the isolation of lactic acid bacteria with antibacterial activity to extend kimchi shelf-life. Antibacterial activity was tested against bacteria associated with acidification of kimchi, including Lactobacillus plantarum, Pediococcus pentosaceus, and Lactobacillus sakei, using agar-well diffusion assay. Two isolates from kimchi were identified as Lactococcus lactis subsp. lactis and Lactobacillus brevis by 16S rRNA gene sequence analysis and API 50 CHL assay, and they showed antibacterial effects against indicator strains. The isolates displayed acid tolerance at pH 3.5, salt tolerance in 5% NaCl, and growth at $4^{\circ}C$. These result imply that the selected strains might be used to extend kimchi shelf-life as a potential starter.

Bacterial Hash Function Using DNA-Based XOR Logic Reveals Unexpected Behavior of the LuxR Promoter

  • Pearson, Brianna;Lau, Kin H.;Allen, Alicia;Barron, James;Cool, Robert;Davis, Kelly;DeLoache, Will;Feeney, Erin;Gordon, Andrew;Igo, John;Lewis, Aaron;Muscalino, Kristi;Parra, Madeline;Penumetcha, Pallavi;Rinker, Victoria G.;Roland, Karlesha;Zhu, Xiao;Poet, Jeffrey L.;Eckdahl, Todd T.;Heyer, Laurie J.;Campbell, A. Malcolm
    • Interdisciplinary Bio Central
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    • v.3 no.3
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    • pp.10.1-10.8
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    • 2011
  • Introduction: Hash functions are computer algorithms that protect information and secure transactions. In response to the NIST's "International Call for Hash Function", we developed a biological hash function using the computing capabilities of bacteria. We designed a DNA-based XOR logic gate that allows bacterial colonies arranged in a series on an agar plate to perform hash function calculations. Results and Discussion: In order to provide each colony with adequate time to process inputs and perform XOR logic, we designed and successfully demonstrated a system for time-delayed bacterial growth. Our system is based on the diffusion of ${\ss}$-lactamase, resulting in destruction of ampicillin. Our DNA-based XOR logic gate design is based on the op-position of two promoters. Our results showed that $P_{lux}$ and $P_{OmpC}$ functioned as expected individually, but $P_{lux}$ did not behave as expected in the XOR construct. Our data showed that, contrary to literature reports, the $P_{lux}$ promoter is bidirectional. In the absence of the 3OC6 inducer, the LuxR activator can bind to the $P_{lux}$ promoter and induce backwards transcription. Conclusion and Prospects: Our system of time delayed bacterial growth allows for the successive processing of a bacterial hash function, and is expected to have utility in other synthetic biology applications. While testing our DNA-based XOR logic gate, we uncovered a novel function of $P_{lux}$. In the absence of autoinducer 3OC6, LuxR binds to $P_{lux}$ and activates backwards transcription. This result advances basic research and has important implications for the widespread use of the $P_{lux}$ promoter.

Antibacterial Activities of Suaeda maritima Extract. (제주도 해홍나물(Suaeda maritima) 추출물의 항균활성)

  • Moon, Young-Gun;Song, Chang-Young;Yeo, In-Kyu;Kim, Gi-Young;Heo, Moon-Soo
    • Journal of Life Science
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    • v.18 no.6
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    • pp.776-781
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    • 2008
  • To develop natural food preservatives, methanol and water extracts were prepared from the Suaeda maritima and their antibacterial activities were examined against 12 microorganisms which were food borne pathogens bacteria, food poisoning microorganisms and food-related bacteria. Methanol extracts exhibited antibacterial activities for the 5 Gram positve and 7 Gram negative bacteria by agar diffusion method, The antibacterial activities and cell growth inhibition were investigated on each strain with the different concentrations of Suaeda maritima extracts. Antibacterial activities were shown in root, stem, furit extracts of Suaeda maritima. However stem and fruit extracts showed weak antibacterial activity against the tested microorganisms. Root extracts showed the highest antibacterial activities against microorganisms tested, such as Bacillus cereus,Bacillus subtilis, Vibrio parahaemolyticus, Staphylococcus aureus. The highest antibacterial activity against bacteria test was found in the methanol extract.

Molecular Characteristics of Extended-Spectrum β-Lactamase Genes in Clinical Isolates of Escherichia coli and Klebsiella pneumoniae (임상검체에서 분리된 Escherichia coli와 Klebsiella pneumoniae의 Extended-Spectrum β-Lactamase 유전자형 및 분자유전학적 특성)

  • Chung, Kyung-Suk
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.1
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    • pp.26-33
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    • 2006
  • Recently, the rapid increase in extended-spectrum ${\beta}$-lactamase (ESBL) producing clinical isolates has become a serious problem. In this study, the epidemiologic features and molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae, antibiotic susceptibility testing, genotype of the ESBL and patterns of chromosomal DNA from PFGE (pulsed field gel electrophoresis) were observed. A total of 53 ESBL-producing clinical isolates (30 of E. coli and 23 of Klebsiella pneumoniae) were collected from two university hospitals in the period of June to July in 2002 and 2003 respectively. The antibiotic resistance frequency of those 53 strains was tested by the disk agar diffusion method with the result that all the strains were resistant to cephalothin. To other antibiotics, the resistance rates of E. coli (30 isolates) were in order of ceftazidime (90.0%), cefotaxime and aztreonam (respectively 83.3%). Also, the resistance rates of K. pneumoniae (23 isolates) were in order of aztreonam (78.3%), ceftazidime (73.9%) and cefotaxime (65.3%). Also the sensitivity of ceftazidime-clavulanic acid were 100% in E. coli and 95.7% in K. pneumoniae. And the sensitivity of cefotaxime-clavulanic acid was 96.7% in E. coli and 91.3% in K. pneumoniae. The types of the ESBL genes were determined by using polymerase chain reaction (PCR). Among the 30 isolates of ESBL-producing E. coli, 6 (20.0%) have SHV only, 5 (16.7%) have TEM only and, 18 (60.0%) have both of TEM and SHV. Among the 23 isolates of ESBL-producing K. pneumoniae, 7 (30.4%) have SHV only, 2 (8.7%) have TEM only, and 14 (60.9%) have both of TEM and SHV. These results show that 52 strains, with only one exception, were confirmed as either TEM or SHV. The patterns of Xba I-digested chromosomal DNA of ESBL-producing E. coli and K. pneumoniae isolates were analyzed by PFGE. PFGE patterns of E. coli and K. pneumoniae were multiclonal, but many strains were grouped into a few types. Therefore, it seems that there were clonal outbreaks or possible horizontal spread. In conclusion, the TEM and SHV ${\beta}$-lactamase are most widely spread in E. coli and K. pneumoniae in Korea. As these types are usually carried by plasmids, the spread of these ${\beta}$-lactamase genes could compromise the future usefulness of third generation cephalosporins for the treatment of infections caused by E. coli and K. pneumoniae.

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Antimicrobial Activity of Psoralea corylifolia, Schisandra chinensis, and Spatholobus suberectus Extracts (보골지, 오미자, 계혈등 추출물의 항균활성)

  • Choi, Eun-Jeong;Jang, Sung-Ran;Kang, Ok-Ju;Bang, Woo-Suk
    • Korean Journal of Food Science and Technology
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    • v.45 no.4
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    • pp.495-500
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    • 2013
  • This study was performed to identify safe natural food preservatives from medicinal herbs and to evaluate the antimicrobial effects of medicinal plants against microorganisms. Medicinal herbs were extracted 3 times with methanol at $45^{\circ}C$ for 3 h and fractionated with n-hexane. The antimicrobial effects of the fractions were determined by measuring the diameter of the inhibition zone by using an agar-well diffusion assay. The MIC of fractions for the inhibition of microorganisms was determined using a microplate reader. The antimicrobial effects of fractions were greater against gram positive bacteria than against gram negative bacteria, but the difference was not significant. The antimicrobial effects of fractions were concentration dependent. While these results have implications, the underlying mechanisms of microbial inactivation need to be further elucidated. The results showed the possibility of developing safer food preservatives.

Identification of Bean Common Mosaic Virus Obtained from Seeds of Phaseolus vulgaris (강낭콩에서 종자전염된 Bean Common Mosaic Virus의 분류동정에 관한 연구)

  • Choi Y.M.;Lee S.H.;Park J.S.;Kim J.S.
    • Korean journal of applied entomology
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    • v.23 no.1 s.58
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    • pp.15-21
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    • 1984
  • The virus infecting French bean (Phaseolus vulgaris L.) was identified as Bean Common Mosaic Virus(BCMV) based on the host range, symptomatology, serology, morphology of virus particles and inclusion bodies. Isolates of BCMV were obtained from seeds of P. vulgaris collected at Suweon, Jangsu and Jinju in Korea. French bean produced vein clearing, mosaic, stunting and leaf curling. Symptom of Chenopodium quinoa was local lesions on the inoculated leaves, not on the upper leaves. The electron micrograph of the virus from French bean was flexuous approximately 750nm in length. Cylindrical and pinwheel cytoplasmic inclusion bodies were observed in French bean leaf infected by BCMV. BCMV from the French bean was transmitted through seed and green peach aphid, Myzus persicae. The thermal inactivation point was $55\~60^{\circ}C$, dilution end point was $10^{-3}\~10^{-5}$ and longevity in vitro was $2\~3$ days for BCMV from French bean. The isolates of BCMV reacted positively against BCMV antiserum. The extract of BCMV infected bean leaves, Azukibean mosaic virus (AZMV) and Cowpea aphid borne mosaic virus(CaMV) also reacted with BCMV antiserum, however, BCMV and CaMV showed the spur in agar gel diffusion test.

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EFFECT OF POLYPHOSPHATE IN ROOT CANAL SEALERS ON THE GROWTH OF ORAL BACTERIA (Polyphosphate가 함유된 근관충전재가 구강세균의 성장에 미치는 영향)

  • 박석범;최기운;최호영
    • Restorative Dentistry and Endodontics
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    • v.26 no.2
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    • pp.141-152
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    • 2001
  • Eliminating the infecting bacteria of the root canal system and preventing reinfection must be the main objectives of all endodontic works. None of commercially available root canal sealers have the properties of desirable tissue compatibility and strong antibacterial activity. The purpose of this study is to develope an ideal root canal sealer using commercially available polyphosphate (polyP), Calgon, which is known to be antibacterial and safe. For the study. resin type AH26, zinc oxide eugenol type Tubli Seal. Ca(OH)$_2$ type Apexit as base sealers for polyP (0~3%) and para formaldehyde containing N2 as a control base were selected. Specimens (3$\times$4mm) of the sealers were prepared in a 37$^{\circ}C$ incubator for 3 and 10 days and their antibacterial activity against streptococci and black pigmented anaerobic rods was observed using an agar diffusion method. The result were as follows: 1. Among 3 day old root canal sealers. N2 as a positive control showed the strongest antibacterial effect. followed by AH26. Tubli Seal and. Apexit which barely showed antibacterial activity against the test bacteria. In contrast. 10 day old AH26 showed a greater antibacterial activity than 10 day old N2. 2. All sealer specimens showed a greater antibacterial activity against black pigmented anaerobic rods than streptococci. Three day old ones appeared to be more antibacterial than 10 day old ones except for Apexit. 3. As compared to N2, 3 day old AH26 demonstrated a similar antibacterial activity against black pig mented anaerobic rods but to a lesser extent to streptococci. Ten day old AH26 showed a greater antibacterial activity against black pigmented anaerobic rods than 10 day old N2. 4. As compared to AH26. Tubli Seal generally revealed a lower antibacterial activity but it showed a greater antibacterial activity aginst S. gordonii Challis. 5. Enhancement of antibacterial activity by polyP was more clearly observed when it was added to Ca(OH)$^{\circ}C$ based root canal sealers. Tubli Seal and N2. 6. The addition of polyP enhanced the antibacterial activity of 3 day old AH26 against S. gordonii G9B (16%) and Challis (29%), and P. gingivalis 2561 (24%) only. Moreover, polyP failed to increase antibacterial activity of 10 day old AH26 against the test strains but P. gingivalis A7A1 28(13%). 7. The addition of polyP increased the antibacterial effect of 3 day old Tubli Seal on several test bacteria including s. mutans GS 5 (50%). s. gordonii G9B (47%) and Challis (122%). and all the test strains of P. gingivalis (13~35%) except for 9 14K 1. The addition of polyP to 10 day old Tubli Seal increased antibacterial activity of the root canal sealer against most test strains. 8. 3 day old Apexit failed to show antibacterial activity. if any very little against S. mutans GS 5 and Pr. intermedia ATCC 49046. However. polyP increased its antibacterial activity by 50 and 69%, respectively. Increase of antibacterial activity of 10 day old Apexit by polyP was more clearly observed than that of 3 day old one.

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Antifungal Activities of the Essential Oils in Syzygium aromaticum (L.) Merr. Et Perry and Leptospermum petersonii Bailey and their Constituents against Various Dermatophytes

  • Park, Mi-Jin;Gwak, Ki-Seob;Yang, In;Choi, Won-Sil;Jo, Hyun-Jin;Chang, Je-Won;Jeung, Eui-Bae;Choi, In-Gyu
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.460-465
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    • 2007
  • This study was carried out in order to investigate the potential of using plant oils derived from Leptospermum petersonii Bailey and Syzygium aromaticum L. Merr. Et Perry as natural antifungal agents. The antifungal effects of essential oils at concentrations of 0.05, 0.1, 0.15, and 0.2 mg/ml on the dermatophytes Microsporum canis (KCTC 6591), Trichophyton mentagrophytes (KCTC 6077), Trichophyton rubrum (KCCM 60443), Epidermophyton floccosum (KCCM 11667), and Microsporum gypseum were evaluated using the agar diffusion method. The major constituents of the active fraction against the dermatophytes were identified by gas chromatography-mass spectrometry and high-performance liquid chromatography analysis. The antifungal activities of S. aromaticum oil (clove oil) against the dermatophytes tested were highest at a concentration of 0.2mg/ml, with an effectiveness of more than 60%. Hyphal growth was completely inhibited in T. mentagrophytes, T. rubrum, and M. gypseum by treatment with clove oil at a concentration of 0.2 mg/ml. Eugenol was the most effective antifungal constituent of clove oil against the dermatophytes T. mentagrophytes and M. canis. Morphological changes in the hyphae of T. mentagrophytes, such as damage to the cell wall and cell membrane and the expansion of the endoplasmic reticulum, after treatment with 0.11 mg/ml eugenol were observed by transmission electron microscopy (TEM). At a concentration of 0.2 mg/ml, L. petersonii oil (LPO) was more than 90% effective against all of the dermatophytes tested, with the exception of T. rubrum. Geranial was determined to be the most active antifungal constituent of L. petersonii oil. Taken together, the results of this study demonstrate that clove and tea tree oils exhibited significant antifungal activities against the dermatophytes tested in this study.