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Sample records for broth microdilution method

  1. Drug susceptibility testing of Mycobacterium tuberculosis by the broth microdilution method with 7H9 broth

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    Ahmet Yilmaz Coban

    2004-02-01

    Full Text Available In this study, we have evaluated the broth microdilution method (BMM for susceptibility testing of Mycobacterium tuberculosis. A total of 43 clinical isolates of M. tuberculosis and H37Rv as a control strain were studied. All isolates were tested by the proportion method and the BMM for isoniazid (INH, rifampicin (RIF, streptomycin (STR, and ethambutol (ETM. The proportion method was carried out according to the National Committee for Clinical Laboratory Standards (NCCLS on Löwenstein-Jensen (LJ medium. The BMM was carried out using 7H9 broth with 96 well-plates. All strains were tested at 3.2-0.05 µg/ml, 16-0.25 µg/ml, 32-0.5 µg/ml, and 32-0.5 µg/ml concentrations for INH, RIF, STR, and ETM, respectively. When the BMM was compared with the proportion method, sensitivity was 100, 100, 96.9, and 90.2%, while specificity was 100, 85.7, 90.9, and 100% for INH, RIF, STR, and ETM, respectively. The plates were examined 7, 10, 14, and 21 days after incubation. The majority of the result were obtained at 14th days after incubation, while the proportion method result were ended in 21-28 days. According to our results, it may be suggested that the BMM is suitable for early determining of multidrug-resistance-M. tuberculosis strains in developed or developing countries.

  2. Antimicrobial Susceptibility Testing of Acinetobacter spp. by NCCLS Broth Microdilution and Disk Diffusion Methods

    OpenAIRE

    Swenson, Jana M.; Killgore, George E.; Tenover, Fred C.

    2004-01-01

    Although both broth microdilution (BMD) and disk diffusion (DD) are listed by NCCLS as acceptable methods for testing Acinetobacter spp. for antimicrobial susceptibility, few studies have compared the results generated by the two methods. We tested 196 isolates of Acinetobacter spp. from nine U.S. hospitals and from the Centers for Disease Control culture collection by using BMD and DD and clinically appropriate antimicrobial agents. Categorical results for amikacin, ciprofloxacin, gatifloxac...

  3. Standardization of a broth microdilution susceptibility testing method to determine minimum inhibitory concentrations of aquatic bacteria

    DEFF Research Database (Denmark)

    Miller, R.A.; Walker, R.D.; Carson, J.;

    2005-01-01

    -Hinton broth. These QC ranges were accepted by the CLSI/NCCLS Subcommittee on Veterinary Antimicrobial Susceptibility Testing in January 2004. This broth microdilution testing method represents the first standardized method for determining MICs of bacterial isolates whose preferred growth temperatures......A multiple laboratory study was conducted in accordance with the standards established by the Clinical and Laboratory Standards Institute (CLSI), formerly the National Committee for Clinical Laboratory Standards (NCCLS), for the development of quality control (QC) ranges using dilution...... antimicrobial susceptibility testing methods for bacterial isolates from aquatic animal species. QC ranges were established for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 when testing at 22, 28 and 35 degrees C (E. coli only) for 10 different antimicrobial agents...

  4. Comparison of Etest with Modified Broth Microdilution Method for Testing Susceptibility of Aspergillus spp. to Voriconazole

    Science.gov (United States)

    Serrano, M. C.; Morilla, D.; Valverde, A.; Chávez, M.; Espinel-Ingroff, A.; Claro, R.; Ramírez, M.; Mazuelos, E. Martín

    2003-01-01

    We compared the Etest with a broth microdilution method, performed according to a modified National Committee for Clinical Laboratory Standards guideline (M38-A), for determining the in vitro susceptibility of 77 isolates of Aspergillus spp. (26 A. fumigatus, 21 A. flavus, 10 A. terreus, 9 A. niger, 5 A. nidulellus, 4 A. glaucus, and 2 A. flavipes isolates). Overall, there was 92.2% agreement between both methods when Etest MICs were read at 24 h and 83.1% agreement when both methods were read at 48 h. When Etest MICs were read at 24 h, the agreement was >90% for all species tested except for A. fumigatus (84.6%). When Etest MICs were read at 48 h, the agreement ranged from 50 to 100%. The poorest agreement was seen with A. glaucus (50%) and A. fumigatus (65%). Where a discrepancy was observed between Etest and the reference method, the Etest MIC was generally higher. The Etest appears to be a suitable alternative procedure for testing the susceptibility of Aspergillus spp. to voriconazole. PMID:14605181

  5. Antifungal susceptibility testing of vaginal candida isolates: the broth microdilution method

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    Mahmoudi Rad M

    2010-02-01

    Full Text Available "n Normal 0 false false false EN-US X-NONE AR-SA MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:Arial; mso-bidi-theme-font:minor-bidi;} Background: Vulvovaginal candidiasis is a common mucosal infection among immunocompetent, healthy women, and is caused by opportunistic yeasts that belong to genus Candida. In this study, we isolated and identified the Candida species in the vagina of patients who admitted in Gynecology Department of Mahdieh Hospital in Tehran, Iran to evaluate the in vitro activities of fluconazole, miconazole, itraconazole and flucytosine against 191 clinical Candida isolates by the NCCLS microdilution method."n"nMethods: 191 Candida were isolated from vaginal secretions and identified with conventional mycological methods in the diagnosis of Candida species. The identity of all strains was confirmed genotypically by multiplex PCR. In vitro susceptibility testing of vaginal Candida isolates was performed by the NCCLS broth microdilution method. The results were read at 48 h."n"nResults: Most C. albicans isolates (>90% were sensitive in vitro to the antifungal agents tested. Most C. glabrata isolates showed sensitivity to miconazole and then flucytosine while they were more resistant to Itraconazole and fluconazole. Many isolates of C. tropicalis were susceptible to miconazole and then fluconazole. They showed a little resistance to

  6. Comparative Evaluation of FUNGITEST and Broth Microdilution Methods for Antifungal Drug Susceptibility Testing of Candida Species and Cryptococcus neoformans

    OpenAIRE

    Davey, Kate G.; Holmes, Ann D.; Elizabeth M. Johnson; Szekely, Adrien; Warnock, David W.

    1998-01-01

    The FUNGITEST method (Sanofi Diagnostics Pasteur, Paris, France) is a microplate-based procedure for the breakpoint testing of six antifungal agents (amphotericin B, flucytosine, fluconazole, itraconazole, ketoconazole, and miconazole). We compared the FUNGITEST method with a broth microdilution test, performed according to National Committee for Clinical Laboratory Standards document M27-A guidelines, for determining the in vitro susceptibilities of 180 isolates of Candida spp. (50 C. albica...

  7. Standardization of a broth microdilution susceptibility testing method to determine minimum inhibitory concentrations of aquatic bacteria

    DEFF Research Database (Denmark)

    Miller, R.A.; Walker, R.D.; Carson, J.; Coles, M.; Coyne, R.; Dalsgaard, Inger; Gieseker, C.; Hsu, H.M.; Mathers, J.J.; Papapetropoulou, M.; Petty, B.; Teitzel, C.; Reimschuessel, R.

    2005-01-01

    antimicrobial susceptibility testing methods for bacterial isolates from aquatic animal species. QC ranges were established for Escherichia coli ATCC 25922 and Aeromonas salmonicida subsp. salmonicida ATCC 33658 when testing at 22, 28 and 35 degrees C (E. coli only) for 10 different antimicrobial agents...

  8. Determination of minimum inhibitory concentrations of itraconazole, terbinafine and ketoconazole against dermatophyte species by broth microdilution method

    Directory of Open Access Journals (Sweden)

    V K Bhatia

    2015-01-01

    Full Text Available Purpose: Various antifungal agents both topical and systemic have been introduced into clinical practice for effectively treating dermatophytic conditions. Dermatophytosis is the infection of keratinised tissues caused by fungal species of genera Trichophyton, Epidermophyton and Microsporum, commonly known as dermatophytes affecting 20–25% of the world's population. The present study aims at determining the susceptibility patterns of dermatophyte species recovered from superficial mycoses of human patients in Himachal Pradesh to antifungal agents; itraconazole, terbinafine and ketoconazole. The study also aims at determining the minimum inhibitory concentrations (MICs of these agents following the recommended protocol of Clinical and Laboratory Standards Institute (CLSI (M38-A2. Methodology: A total of 53 isolates of dermatophytes (T. mentagrophyte-34 in no., T. rubrum-18 and M. gypseum-1 recovered from the superficial mycoses were examined. Broth microdilution method M38-A2 approved protocol of CLSI (2008 for filamentous fungi was followed for determining the susceptibility of dermatophyte species. Results: T. mentagrophyte isolates were found more susceptible to both itraconazole and ketoconazole as compared to terbinafine (MIC50: 0.125 µg/ml for itraconazole, 0.0625 µg/ml for ketoconazole and 0.5 µg/ml for terbinafine. Three isolates of T. mentagrophytes (VBS-5, VBSo-3 and VBSo-73 and one isolate of T. rubrum (VBPo-9 had higher MIC values of itraconazole (1 µg/ml. Similarly, the higher MIC values of ketoconazole were observed in case of only three isolates of T. mentagrophyte (VBSo-30 = 2 µg/ml; VBSo-44, VBM-2 = 1 µg/ml. The comparative analysis of the three antifungal drugs based on t-test revealed that 'itraconazole and terbinafine' and 'terbinafine and ketoconazole' were found independent based on the P < 0.005 in case of T. mentagrophyte isolates. In case of T. rubrum, the similarity existed between MIC values of 'itraconazole and

  9. Susceptibility testing with the sensititer breakpoint broth microdilution system.

    Science.gov (United States)

    Doern, G V; Dascal, A; Keville, M

    1985-05-01

    The antimicrobial susceptibility profiles of a total of 318 aerobic and facultatively anaerobic bacteria (255 gram-negative bacilli and 63 gram-positive cocci) were determined, using a new commercially available breakpoint broth microdilution procedure (Sensititer Breakpoint System (SBS), Gibco Diagnostics, Inc., Madison, WI) that categorizes test results in the form of susceptibility categories: susceptible, intermediate, and resistant. Results obtained with the SBS were compared with those achieved with a standardized disk diffusion procedure. Among a total of 4,414 organism-antimicrobic comparisons, concordance between the results of the SBS and the disk diffusion procedure was observed in 3,888 cases (88.1%). Four hundred twenty-three (9.6%) minor discrepancies, 45 (1.0%) major discrepancies, and 58 (1.3%) very major discrepancies were noted. Arbitration of major and very major discrepancies with a full-range minimum inhibitory concentration (MIC) procedure confirmed the results of the SBS in 53.4% of cases. A single organism-antimicrobial combination, the nonenterococcal streptococci tested against the aminoglycosides, yielded a significant number of very major errors which were arbitrated in favor of the disk diffusion result. These errors were probably due to poor growth of the test organism in the broth medium used for performing the SBS test (i.e., cation-supplemented Mueller-Hinton broth). With this exception, the SBS was found to be at least as accurate as the standardized disk diffusion procedure. PMID:3922668

  10. Comparative Evaluation of the VITEK 2, Disk Diffusion, Etest, Broth Microdilution, and Agar Dilution Susceptibility Testing Methods for Colistin in Clinical Isolates, Including Heteroresistant Enterobacter cloacae and Acinetobacter baumannii Strains▿

    OpenAIRE

    Lo-Ten-Foe, Jerome R; de Smet, Anne Marie G. A.; Diederen, Bram M. W.; Kluytmans, Jan A J W; van Keulen, Peter H. J.

    2007-01-01

    Increasing antibiotic resistance in gram-negative bacteria has recently renewed interest in colistin as a therapeutic option. The increasing use of colistin necessitates the availability of rapid and reliable methods for colistin susceptibility testing. We compared seven methods of colistin susceptibility testing (disk diffusion, agar dilution on Mueller-Hinton [MH] and Isosensitest agar, Etest on MH and Isosensitest agar, broth microdilution, and VITEK 2) on 102 clinical isolates collected f...

  11. Investigation of susceptibility of Staphylococcus species to some antibacterial drugs by disk diffusion and broth microdilution

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    Ašanin Jelena

    2012-01-01

    Full Text Available The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For primary identification, catalase and oxidase tests were used, as well as the free coagulase test. Following the preliminary tests, the isolated strains were identified using commercial systems ID32 STAPH (bioMérieux, France and the BBL Crystal Gram-Positive ID Kit (Becton Dickinson, USA according to the enclosed instructions. The Staphylococcus isolates were examined for sensitivity to the following: oxacillin, penicillin, cefoxitin, gentamicin, erythromycin, chloramphenicol, tetracycline, ciprofloxacin, sulfametoxazol/trimetoprim, and vacomycin using the disk diffusion method and the broth microdilution method as recommended by the Clinical and Laboratory Strandards Institute - CLSI(2003, and the results were interpreted according to CLSI recommendations from 2008 and 2010. Antibiogram disks manufactured by Becton Dickinson (USA were used, and the broth microdilution method was applied using pure antibiotic substances from different manufacturers: erythromycin, chloramphenicol, cefoxitin, gentamicin, oxacillin, tetracycline (Sigma Aldrich, USA, sulfametoxazol (Fluka, USA, penicillin (Calbiochem, Germany, vancomycin (Abbott laboratories, USA, ciprofloxacin and trimetoprim (Zdravlje A.D., Serbia. All 25 strains were catalase positive and oxidase negative. Of the 25 strains, 19 were coagulase positive and 6 were coagulase negative.With the implementation of the disk diffusion method on 19 strains of S. aureus, 17 were established to be resistant to penicillin (89.5%, and 2 strains to gentamicin

  12. Comparison of the EUCAST and CLSI Broth Microdilution Methods for Testing Isavuconazole, Posaconazole, and Amphotericin B against Molecularly Identified Mucorales Species

    Science.gov (United States)

    Singh, Pradeep Kumar; Kathuria, Shallu; Hagen, Ferry; Meis, Jacques F.

    2015-01-01

    We compared EUCAST and CLSI antifungal susceptibility testing (AFST) methods for triazoles and amphotericin B against 124 clinical Mucorales isolates. The EUCAST method yielded MIC values 1- to 3-fold dilutions higher than those of the CLSI method for amphotericin B. The essential agreements between the two methods for triazoles were high, i.e., 99.1% (voriconazole), 98.3% (isavuconazole), and 87% (posaconazole), whereas it was significantly lower for amphotericin B (66.1%). Strategies for harmonization of the two methods for Mucorales AFST are warranted. PMID:26438489

  13. Detection of amphotericin B resistance in Candida haemulonii and closely related species by use of the Etest, Vitek-2 yeast susceptibility system, and CLSI and EUCAST broth microdilution methods.

    Science.gov (United States)

    Shin, Jong Hee; Kim, Mi-Na; Jang, Sook Jin; Ju, Min Young; Kim, Soo Hyun; Shin, Myung Geun; Suh, Soon Pal; Ryang, Dong Wook

    2012-06-01

    The emerging fungal pathogens Candida haemulonii and Candida pseudohaemulonii often show high-level resistance to amphotericin B (AMB). We compared the utilities of five antifungal susceptibility testing methods, i.e., the Etest using Mueller-Hinton agar supplemented with glucose and methylene blue (Etest-MH), the Etest using RPMI agar supplemented with glucose (Etest-RPG), the Vitek-2 yeast susceptibility system, and the Clinical and Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth microdilution methods, for the detection of AMB-resistant isolates of C. haemulonii and closely related species. Thirty-eight clinical isolates (8 C. haemulonii, 10 C. pseudohaemulonii, and 20 Candida auris isolates) were analyzed. Of the 18 C. haemulonii and C. pseudohaemulonii isolates, 18, 15, 18, 10, and 9 exhibited AMB MICs of >1 μg/ml by the Etest-MH, Etest-RPG, Vitek-2, CLSI, and EUCAST methods, respectively. All 20 C. auris isolates showed AMB MICs of ≤1 μg/ml by all five methods. Of the methods, the Etest-MH generated the broadest distribution of AMB MICs for all 38 isolates and showed the best discrimination between the C. haemulonii and C. pseudohaemulonii isolates (4 to 32 μg/ml) and those of C. auris (0.125 to 0.5 μg/ml). Taking the Etest-MH as the reference method, the essential agreements (within two dilutions) for the Etest-RPG, Vitek-2, CLSI, and EUCAST methods were 84, 92, 55, and 55%, respectively; the categorical agreements were 92, 92, 79, and 76%, respectively. This study provides the first data on the efficacy of the Etest-MH and its excellent agreement with Vitek-2 for discriminating AMB-resistant from AMB-susceptible isolates of these Candida species. PMID:22442324

  14. Comparison of in vitro activity of five antifungal agents against dermatophytes, using the agar dilution and broth microdilution methods Comparação da atividade in vitro de cinco agentes antifúngicos para dermatófitos, usando os métodos de diluição em ágar e microdiluição em caldo

    OpenAIRE

    Crystiane Rodrigues Araújo Mota; Karla Carvalho Miranda; Janine de Aquino Lemos; Carolina Rodrigues Costa; Souza, Lúcia Kioko Hasimoto e; Xisto Sena Passos; Hildene Meneses e Silva; Maria do Rosário Rodrigues Silva

    2009-01-01

    The purpose of this study was to compare the agar dilution and broth microdilution methods for determining the minimum inhibitory concentration (MIC) of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine for 60 dermatophyte samples belonging to the species Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The percentage agreement between the two methods, for all the isolates with < 2 dilutions that were tested was 91.6% for ketoconazole and griseofulvi...

  15. Sensititre autoreader for same-day breakpoint broth microdilution susceptibility testing of members of the family Enterobacteriaceae.

    OpenAIRE

    Doern, G V; Staneck, J L; Needham, C.; Tubert, T

    1987-01-01

    The Sensititre Autoreader system is an instrument-assisted broth microdilution susceptibility test procedure based on the detection of fluorogenic growth substrate metabolism by test bacteria with different concentrations of antimicrobial agents. In the current investigation, this system was assessed as a means for predicting the in vitro activity of 17 antimicrobial agents versus numerous species of the family Enterobacteriaceae and Pseudomonas aeruginosa by using a breakpoint broth microdil...

  16. Antibiotic susceptibility of members of the Lactobacillus acidophilus group using broth microdilution and molecular identification of their resistance determinants

    NARCIS (Netherlands)

    Mayrhofer, S.; Hoek, van A.H.A.M.; Mair, C.; Huys, G.; Aarts, H.J.M.; Kneifel, W.; Domig, K.J.

    2010-01-01

    The range of antibiotic susceptibility to 13 antibiotics in 101 strains of the Lactobacillus acidophilus group was examined using the lactic acid bacteria susceptibility test medium (LSM) and broth microdilution. Additionally, microarray analysis and PCR were applied to identify resistance genes res

  17. Determination of Susceptibilities of 26 Leptospira sp. Serovars to 24 Antimicrobial Agents by a Broth Microdilution Technique

    OpenAIRE

    Murray, Clinton K.; Hospenthal, Duane R.

    2004-01-01

    The MICs of 24 antimicrobials for 26 Leptospira spp. serovars were determined using a broth microdilution technique. The MICs at which 90% of isolates tested were inhibited (MIC90s) of cefepime, imipenem-cilastatin, erythromycin, clarithromycin, and telithromycin were all ≤0.01 μg/ml. The MIC90s of amoxicillin, aztreonam, cefdinir, chloramphenicol, and penicillin G were ≥3.13 μg/ml. Many antimicrobials have excellent in vitro activity against Leptospira.

  18. Comparison of in vitro activity of five antifungal agents against dermatophytes, using the agar dilution and broth microdilution methods Comparação da atividade in vitro de cinco agentes antifúngicos para dermatófitos, usando os métodos de diluição em ágar e microdiluição em caldo

    Directory of Open Access Journals (Sweden)

    Crystiane Rodrigues Araújo Mota

    2009-06-01

    Full Text Available The purpose of this study was to compare the agar dilution and broth microdilution methods for determining the minimum inhibitory concentration (MIC of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine for 60 dermatophyte samples belonging to the species Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The percentage agreement between the two methods, for all the isolates with O propósito do presente trabalho foi comparar os métodos de diluição em ágar e diluição em caldo para a determinação de concentração inibitória mínima de fluconazol, itraconazol, cetoconazol, griseofulvina e terbinafina para 60 amostras de dermatófitos pertencentes às espécies, Trichophyton rubrum, Trichophyton. mentagrophytes e Microsporum canis. A porcentagem de acordo entre os dois métodos para todos os isolados testados considerando-se valores < 2 diluições, foram de 91,6% para cetoconazol e para griseofulvina, de 88,3% para itraconazol, de 81,6% para terbinafina e de 73,3% para fluconazol. Uma concordância de 100% foi obtido para isolados de Trichophyton mentagrophytes avaliados com cetoconazol e griseofulvina. Desta forma, até que um método de referência seja padronizado para testar a suscetibilidade in vitro para os dermatófitos, os resultados semelhantes encontrados para os dois métodos fazem com que o método de diluição em ágar possa ser útil no teste de suscetibilidade para estes fungos filamentosos.

  19. Antibiotic susceptibility of members of the Lactobacillus acidophilus group using broth microdilution and molecular identification of their resistance determinants.

    Science.gov (United States)

    Mayrhofer, Sigrid; van Hoek, Angela H A M; Mair, Christiane; Huys, Geert; Aarts, Henk J M; Kneifel, Wolfgang; Domig, Konrad J

    2010-11-15

    The range of antibiotic susceptibility to 13 antibiotics in 101 strains of the Lactobacillus acidophilus group was examined using the lactic acid bacteria susceptibility test medium (LSM) and broth microdilution. Additionally, microarray analysis and PCR were applied to identify resistance genes responsible for the displayed resistant phenotypes in a selection of strains. In general, narrow as well as broad unimodal and bimodal MIC distributions were observed for the Lactobacillus acidophilus group and the tested antimicrobial agents. Atypically resistant strains could be determined by visual inspection of the obtained MIC ranges for ampicillin, chloramphenicol, clindamycin, erythromycin, quinupristin/dalfopristin, streptomycin and tetracycline. For most of these atypically resistant strains underlying resistance determinants were found. To our knowledge erm(A) was detected in lactobacilli for the first time within this study. Data derived from this study can be used as a basis for reviewing present microbiological breakpoints for categorization of susceptible and resistant strains within the Lactobacillus acidophilus group to assess the safety of microorganisms intended for use in food and feed applications. PMID:20888656

  20. Dermatophyte susceptibilities to antifungal azole agents tested in vitro by broth macro and microdilution methods Suscetibilidade in vitro de dermatófitos a azóis pelos métodos macro e microdiluição em caldo

    Directory of Open Access Journals (Sweden)

    Emerson Roberto Siqueira

    2008-02-01

    Full Text Available The in vitro susceptibility of dermatophytes to the azole antifungals itraconazole, fluconazole and ketoconazole was evaluated by broth macro and microdilution methods, according to recommendations of the CLSI, with some adaptations. Twenty nail and skin clinical isolates, four of Trichophyton mentagrophytes and 16 of T. rubrum were selected for the tests. Itraconazole minimal inhibitory concentrations (MIC varied from Foi avaliada a suscetibilidade in vitro de dermatófitos aos antifúngicos itraconazol, fluconazol e cetoconazol, pelos métodos macro e microdiluição em caldo, de acordo com as recomendações do CLSI, com algumas modificações. Foram estudados 20 isolados clínicos de lesões de unha e pele, sendo quatro Trichophyton mentagrophytes e 16 T. rubrum. A concentração inibitória mínima (CIM para itraconazol variou de < 0,03 a 0,25 µg/mL pelo método da macrodiluição, e de < 0,03 a 0,5 µg/mL pela microdiluição em caldo; de 0,5 a 64 µg/mL e de 0,125 a 16 µg/mL para fluconazol, respectivamente, pela macro e microdiluição; e de < 0,03 a 0,5 µg/mL por ambos os métodos para cetoconazol. A concordância entre os dois métodos (considerando ± uma diluição foi de 70% para itraconazol, 45% para fluconazol e 85% para cetoconazol. Conclui-se que os isolados estudados foram inibidos por concentrações relativamente baixas dos antifúngicos testados, e os dois métodos apresentam boa concordância, especialmente para itraconazol e cetoconazol.

  1. Avaliação das metodologias M.I.C.E.®, Etest® e microdiluição em caldo para determinação da CIM em isolados clínicos Evaluation of M.I.C.E.TM, Etest® and CLSI broth microdilution methods for antimicrobial susceptibility testing of nosocomial bacterial isolates

    Directory of Open Access Journals (Sweden)

    Eloiza Helena Campana

    2011-04-01

    .I.C.E., Thermo Fisher Scientific, Basingstoke, UK, recently released into the market, represents a rapid alternative to antimicrobial susceptibility testing. OBJECTIVE: The objective of this study was to evaluate the performance of M.I.C.E. methodology in relation to broth microdilution (reference test and Etest® (BioMérieux, Marcy l'Étoile, France. Material and method: A total of 160 bacterial isolates were collected comprising the following species: P. aeruginosa (20, Acinetobacter spp. (20, K. pneumoniae (20, E. coli (20, S. aureus (20, coagulase-negative Staphylococcus (20, E. faecalis (20 and E. faecium (20. Following Clinical Laboratory Standands Institute (CLSI standards (2009 and the manufacturers' recommendations, antimicrobial susceptibility testing was performed using broth microdilution method, Etest and M.I.C.E. The results were interpreted according to the criteria established by CLSI and compared through regression analysis. RESULTS: All antimicrobial combinations vs. bacterial species were evaluated and M.I.C.E. methodology yielded good results with general correlation (MIC variation ± 1-log2 > 90%, except for cefotaxime (85% and vancomycin (76.3% when compared with the reference method. The M.I.C.E. results compared to Etest showed general correlation (> 96%, except for amoxicillin/clavulanic acid (67.5% combination. CONCLUSION: AST results obtained from M.I.C.E. methodology showed a good correlation with those from broth microdilution and Etest, which corroborates its time effectiveness in the determination of MIC. However, the combination of amoxicillin/clavulanic acid requires further attention.

  2. Quality control ranges for testing broth microdilution susceptibility of Flavobacterium columnare and F. psychrophilum to nine antimicrobials

    DEFF Research Database (Denmark)

    Gieseker, Charles M.; Mayer, Tamara D.; Crosby, Tina C.;

    2012-01-01

    salmonicida subsp. salmonicida ATCC 33658 against 10 antimicrobials (ampicillin, enrofloxacin, erythromycin, florfenicol, flumequine, gentamicin, ormetoprim/sulfadimethoxine, oxolinic acid, oxytetracycline, and trimethoprim/sulfamethoxazole) in diluted (4 g l−1) cation-adjusted Mueller-Hinton broth incubated...

  3. In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method Teste de suscetibilidade in vitro de dermatófitos isolados em Goiânia, Brasil, contra cinco agentes antifúngicos pelo método de microdiluição em caldo

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    Crystiane Rodrigues Araújo

    2009-02-01

    Full Text Available The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 ºC allowing a reading of the minimal inhibitory concentration (MIC after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 µg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.Atividades antifúngicas de fluconazol, itraconazol, cetoconazol, terbinafina e griseofulvina foram testadas pelo método de microdiluição em caldo contra 60 isolados de dermatófitos. Os resultados mostraram que todos os isolados produziram crescimento claramente detectável a 28 ºC e a concentração inibitória mínima (CIM foi determinada após quatro dias de incubação para Trichophyton mentagrophytes e cinco dias para T. rubrum e Microsporum canis. A maioria dos isolados teve um padrão uniforme de suscetibilidade para os agentes antifúngicos testados. Baixos valores de CIM como 0,03 µg/mL foram encontrados para 33,3%, 31,6% e 15% dos isolados para itraconazol, cetoconazol e terbinafina, respectivamente.

  4. Comparison of the serum-supplemented Todd-Hewitt and the new Haemophilus test media for broth microdilution susceptibility testing of Streptococcus pneumoniae.

    Science.gov (United States)

    Roger, M; Lapointe, J R

    1996-06-01

    Horse serum-supplemented Todd-Hewitt broth (STH) in use at Hôpital Ste-Justine for the last 12 years was compared to the recently proposed Haemophilus test medium (HTM), for broth microdilution susceptibility testing of Streptococcus pneumoniae. One hundred and twenty S. pneumoniae isolates from pediatric clinical specimens were used in this study. In general, the minimum inhibitory concentrations (MICs) in STH for 15 antimicrobial agents were quite comparable to those determined in HTM but tended to be higher. Drugs which generated MICs within +/- 1 log2 concentration differences in both media included penicillin, ampicillin, oxacillin, cefuroxime, cefotaxime, cefixime, clindamycin, chloramphenicol, trimethoprim-sulfamethoxazole, rifampin, ciprofloxacin and vancomycin. Cefaclor and tetracycline MICs tended to be > or = 2 log2 dilutions higher with STH for most of the isolates tested, while erythromycin MICs were often 2 log2 dilutions lower with STH than with HTM. Despite some differences in MICs noted above, few very major (0.4%), major (0.2%) and minor interpretive category errors (4.4%) were observed. The visual reading of the MICs for most of the 120 clinical isolates tested was generally easier in STH which was superior in supporting best the bacterial growth as detected by spectrophotometry. The risk of false susceptibility is thus decreased by using STH rather than HTM; furthermore, STH is free of the technical problems of the lysed horse blood Mueller-Hinton (LHB-MH) recommended by the NCCLS. PMID:8808713

  5. [Assessment of 2 automated microdilution techniques compared to an agar dilution method in determining sensitivity to fosfomycin in strains of carbapenem-resistant Pseudomonas aeruginosa].

    Science.gov (United States)

    Gil-Romero, Yolanda; Regodón-Domínguez, Marta; Wilhelmi de Cal, Isabel; López-Fabal, Fátima; Gómez-Garcés, José Luis

    2016-01-01

    Carbapenems-resistance in Pseudomonas aeruginosa isolates has been widely reported. Fosfomycin has been shown to act synergistically with other antimicrobials. The agar dilution method was approved for susceptibility testing for fosfomycin and Pseudomonas aeruginosa. However, broth microdilution methods are the basis of systems currently used in clinical microbiology laboratories. The results of this study indicate that these methods are acceptable as susceptibility testing methods for fosfomycin against these organisms. PMID:26620604

  6. Comparison between E-test and CLSI broth microdilution method for antifungal susceptibility testing of Candida albicans oral isolates Comparação entre E-test e o método da microdiluição do CLSI para teste de susceptibilidade a antifúngicos de isolados orais de Candida albicans

    Directory of Open Access Journals (Sweden)

    Cristiane Yumi Koga-Ito

    2008-02-01

    Full Text Available Thirty Candida albicans isolated from oral candidosis patients and 30 C. albicans isolated from control individuals were studied. In vitro susceptibility tests were performed for amphotericin B, fluconazole, 5-flucytosine and itraconazole through the Clinical and Laboratorial Standards Institute (CLSI reference method and E test system. The results obtained were analyzed and compared. MIC values were similar for the strains isolated from oral candidosis patients and control individuals. The agreement rate for the two methods was 66.67% for amphotericin B, 53.33% for fluconazole, 65% for flucytosine and 45% for itraconazole. According to our data, E test method could be an alternative to trial routine susceptibility testing due to its simplicity. However, it can not be considered a substitute for the CLSI reference method.Trinta Candida albicans isoladas de pacientes portadores de candidose oral e 30 Candida albicans isoladas de indivíduos controle foram estudadas. Testes de susceptibilidade in vitro foram realizados com anfotericina B, fluconazol, 5-flucitosina e itraconazol pelo método do Clinical and Laboratorial Standars Institute (CLSI e por E-test. Os resultados obtidos foram analisados e comparados. Os valores de CIM foram semelhantes para amostras isoladas de pacientes portadores de candidose oral e indivíduos controle. A concordância entre os dois métodos foi de 66,7% para a anfotericina B, 53,33% para o fluconazol, 65% para a flucitosina e 45% para o itraconazol. De acordo com estes resultados, o método do E-test poderia ser uma alternativa para a triagem de casos de rotina pela sua simplicidade. Entretanto, este método não pode ser considerado como um substituto para o método de referência do CLSI.

  7. In vitro effect of intracanal medicaments on strict anaerobes by means of the broth dilution method

    Directory of Open Access Journals (Sweden)

    ROSA Odila Pereira da Silva

    2002-01-01

    Full Text Available The determination of bacterial susceptibility to intracanal medicaments is a necessity. Nevertheless, few studies utilize the proper methodology to carry out that evaluation with anaerobes. In this study, the steps of a broth dilution method, carried out in microplates (microdilution and tubes (macrodilution, to test the effect of traditional intracanal medicaments on anaerobic bacteria are described. The results are presented as values of minimal inhibitory and bactericidal concentrations (MIC and MBC. Standardized inocula of the anaerobic bacteria Prevotella nigrescens (ATCC 33563, Fusobacterium nucleatum (ATCC 25586 and Clostridium perfringens (ATCC 13124, in reinforced Clostridium medium (RCM and supplemented Brucella broth, were submitted to different concentrations of calcium hydroxide, chlorhexidine digluconate, camphorated paramonochlorophenol and formocresol solutions. The drugs were diluted in the same culture broths, in microplates and tubes, and were then incubated in anaerobiosis jars at 37ºC for 48 or 96 hours. The determination of MICs was carried out through visual and spectrophotometric readings, and the determination of MBCs, through the plating of aliquots on RCM-blood agar. For that kind of study, the macromethod with spectrophotometric reading should be the natural choice. MICs and MBCs obtained with the macromethod were compatible with the known clinical performance of the studied medications, and the values varied according to the bacteria and culture media employed. RCM was the most effective medium and C. perfringens, the most resistant microorganism.

  8. Use of Fatty Acid RPMI 1640 Media for Testing Susceptibilities of Eight Malassezia Species to the New Triazole Posaconazole and to Six Established Antifungal Agents by a Modified NCCLS M27-A2 Microdilution Method and Etest

    OpenAIRE

    Velegraki, Aristea; Alexopoulos, Evangelos C.; Kritikou, Stavroula; Gaitanis, George

    2005-01-01

    A novel formulation of RPMI 1640 medium for susceptibility testing of Malassezia yeasts by broth microdilution (BMD) and Etest is proposed. A modification of the NCCLS M27-A2 BMD method was used to test 53 isolates of Malassezia furfur (12 isolates), M. sympodialis (8 isolates), M. slooffiae (4 isolates), M. globosa (22 isolates), M. obtusa (2 isolates), M. restricta (2 isolates), M. pachydermatis (1 isolates), and M. dermatis (2 isolates) against amphotericin B, ketoconazole, itraconazole, f...

  9. Comparative study of disk diffusion and microdilution methods for evaluation of antifungal activity of natural compounds against medical yeasts Candida spp and Cryptococcus sp

    Directory of Open Access Journals (Sweden)

    L. SCORZONI

    2009-01-01

    Full Text Available

    Antifungal activity of natural products has been tested by adapting methods designed for synthetic drugs. In this study, two methods for the determination of antifungal activity of natural products, agar diffusion and broth microdilution, the CLSI reference methods for synthetic drugs, are compared and discussed. The microdilution method was more sensitive. The minimal inhibitory concentrations (MIC of crude extracts, fractions and pure substances from different species of the plant families Piperaceae, Rubiaceae, Clusiaceae, Fabaceae and Lauraceae, from the Biota project, were determined. Antifungal activities against Candida albicans, C.krusei, C.parapsilosis and Cryptococcus neoformans were produced by several samples. Keywords: natural products; antifungal activity; Candida sp; Cryptococcus sp.

  10. Telavancin demonstrates activity against methicillin-resistant Staphylococcus aureus isolates with reduced susceptibility to vancomycin, daptomycin, and linezolid in broth microdilution MIC and one-compartment pharmacokinetic/pharmacodynamic models.

    Science.gov (United States)

    Smith, Jordan R; Barber, Katie E; Hallesy, Jessica; Raut, Animesh; Rybak, Michael J

    2015-09-01

    Methicillin-resistant Staphylococcus aureus (MRSA) isolates have arisen with reduced susceptibility to several anti-MRSA agents. Telavancin (TLV), a novel anti-MRSA agent, retains low MICs against these organisms. Our objective was to determine the MICs for TLV, daptomycin (DAP), vancomycin (VAN), and linezolid (LZD) against daptomycin-nonsusceptible (DNS) S. aureus, vancomycin-intermediate S. aureus (VISA), heteroresistant VISA (hVISA), and linezolid-resistant (LZD(r)) S. aureus. We also evaluated these agents against each phenotype in pharmacokinetic/pharmacodynamic (PK/PD) models. Seventy DNS, 100 VISA, 180 hVISA, and 25 LZD(r) MRSA isolates were randomly selected from our library and tested to determine their MICs against TLV, DAP, VAN, and LZD via broth microdilution and a Trek panel. Four isolates were randomly selected for 168-h in vitro models to evaluate treatment with TLV at 10 mg/kg of body weight/day, DAP at 10 mg/kg/day, VAN at 1 g every 12 h (q12h), and LZD at 600 mg q12h. The MIC50/90 for TLV, DAP, VAN, and LZD against 70 DNS S. aureus isolates were 0.06/0.125 μg/ml, 2/4 μg/ml, 1/2 μg/ml, and 2/2 μg/ml, respectively. Against 100 VISA isolates, the MIC50/90 were 0.06/0.125 μg/ml, 1/1 μg/ml, 4/8 μg/ml, and 1/2 μg/ml, respectively. Against 170 hVISA isolates, the MIC50/90 were 0.06/0.125 μg/ml, 0.5/1 μg/ml, 1/2 μg/ml, and 1/2 μg/ml, respectively. Against 25 LZD(r) isolates, the MIC50/90 were 0.03/0.06 μg/ml, 1/1 μg/ml, 2/2 μg/ml, and 8/8 μg/ml, respectively. The TLV MIC was >0.125 μg/ml for 10/365 (2.7%) isolates. In PK/PD models, TLV was universally bactericidal at 168 h and statistically superior to all antibiotics against DNS S. aureus strain R2334. These data further establish the potency of TLV against resistant MRSA. The model data demonstrate in vitro bactericidal activity of TLV against hVISA, VISA, DNS S. aureus, and LZD(r) S. aureus strains. Further clinical research is warranted. PMID:26124162

  11. Comparative performance of isolation methods using Preston broth, Bolton broth and their modifications for the detection of Campylobacter spp. from naturally contaminated fresh and frozen raw poultry meat.

    Science.gov (United States)

    Seliwiorstow, T; De Zutter, L; Houf, K; Botteldoorn, N; Baré, J; Van Damme, I

    2016-10-01

    The performance of different isolation methods was evaluated for the detection of Campylobacter from naturally contaminated raw poultry meat. Therefore, fresh and frozen poultry meat samples were analysed using the standard procedure (ISO 10272-1:2006), enrichment in Preston broth, and enrichment in modified Bolton broth (supplemented with (i) potassium clavulanate (C-BB), (ii) triclosan (T-BB), (iii) polymyxin B (P-BB)). The enrichment cultures were streaked onto both modified charcoal cefoperazone deoxycholate agar (mCCDA) and RAPID'Campylobacter agar (RCA). Moreover, direct plating on mCCDA and RCA was performed to quantify Campylobacter. In total, 33 out of 59 fresh retail meat samples (55.9%) were Campylobacter positive. For both fresh and frozen poultry meat samples, enrichment in Bolton broth (ISO 10272-1:2006) resulted in a higher number of positive samples than enrichment in Preston broth. Supplementation of Bolton broth with potassium clavulanate (C-BB) and triclosan (T-BB) enhanced the Campylobacter recovery from fresh poultry meat compared to non-supplemented Bolton broth, although the use of C-BB was less applicable than T-BB for Campylobacter recovery from frozen samples. Additionally, the use of RCA resulted in a higher isolation rate compared to mCCDA. The present study demonstrates the impact of culture medium on the recovery of Campylobacter from fresh and frozen naturally contaminated poultry meat samples and can support laboratories in choosing the most appropriate culturing method to detect Campylobacter. PMID:27391222

  12. Modified broth-disk method for testing the antibiotic susceptibility of anaerobic bacteria.

    Science.gov (United States)

    Wilkins, T D; Thiel, T

    1973-03-01

    The most commonly used method for testing the antibiotic susceptibility of aerobic and facultative bacteria is the disk diffusion method. However, some anaerobic bacteria do not grow well enough in anaerobic jars for performance of disk diffusion tests. A modification of the broth-disk method of Schneierson allowed us to determine antibiotic susceptibility in a completely anaerobic environment. Commercial antibiotic disks were added anaerobically to tubes of prereduced brain heart infusion broth to achieve a concentration of each antibiotic approximating that attainable in blood. The tubes were then inoculated and incubated for 18 h. Resistance or susceptibility to each antibiotic was determined according to the amount of growth in each tube as compared with a control culture without the antibiotic. There was good correlation between results obtained by this broth-disk method and minimal inhibitory concentrations. PMID:4790595

  13. Validation of a HILIC Method for the Analysis of Ergothioneine in Fermentation Broth.

    Science.gov (United States)

    Liu, Qi; Zhang, Weiya; Wang, Hongyu; Li, Yunhua; Liu, Wei; Wang, Qunjie; Liu, Dongze; Chen, Ning; Jiang, Wenxia

    2016-07-01

    A hydrophilic interaction liquid chromatography method has been established for the quantification of ergothioneine (EGT) in fermentation broth. Chromatographic separation was conducted on a Venusil hydrophilic interaction liquid chromatography (HILIC) column (250 × 4.6 mm, 5 µm) at an elution rate of 1.0 mL/min with an isocratic mobile phase consisting of acetonitrile/20 mmol/L ammonium acetate solution (85 : 15, v/v) adjusted to pH 6.0 with acetic acid. Analytes were detected at 254 nm using a UV-VIS detector. The injection volume was 10 µL, and the column temperature was 40°C. The limits of detection and limits of quantification were 63 and 21 µg/L, respectively. Excellent linearity [correlation coefficient (R(2)) = 0.9999] was achieved for EGT quantification in the range of 5-400 mg/L. The relative standard deviations of repeatability, intermediate precision and stability were 1.47, 1.03 and 1.66%, respectively, and EGT recoveries were within 99.2-100.8%. The chromatographic peak corresponding to EGT in the HILIC spectrum was confirmed using ESI-MS. In general, the method developed here is simple, reliable, accurate, and stable and may be useful for routine analyses in EGT biosynthesis research. PMID:26921894

  14. An improved HPLC-DAD method for clavulanic acid quantification in fermentation broths of Streptomyces clavuligerus.

    Science.gov (United States)

    Ramirez-Malule, Howard; Junne, Stefan; López, Carlos; Zapata, Julian; Sáez, Alex; Neubauer, Peter; Rios-Estepa, Rigoberto

    2016-02-20

    Clavulanic acid (CA) is an important secondary metabolite commercially produced by cultivation of Streptomyces clavuligerus (Sc). It is a potent inhibitor of bacterial β-lactamases. In this work, a specific and improved high performance liquid chromatography (HPLC) method, using a C-18 reversed phase column, diode array detector and gradient elution for CA quantification in fermentation broths of Sc, was developed and successfully validated. Samples were imidazole-derivatized for the purpose of creating a stable chromophore (clavulanate-imidazole). The calibration curve was linear over a typical range of CA concentration between 0.2 and 400mg/L. The detection and quantification limits were 0.01 and 0.02mg/L, respectively. The precision of the method was evaluated for CA spiked into production media and a recovery of 103.8%, on average, was obtained. The clavulanate-imidazole complex was not stable when the samples were not cooled during the analysis. The recovery rate was 39.3% on average. This assay was successfully tested for CA quantification in samples from Sc fermentation, using both, a chemically defined and a complex medium. PMID:26760242

  15. Indirect methods for characterization of carbon dioxide levels in fermentation broth.

    Science.gov (United States)

    Frick, R; Junker, B

    1999-01-01

    Various factors which influence dissolved carbon dioxide levels were indirectly evaluated in pilot scale and laboratory studies. For pilot scale studies, off-gas carbon dioxide (percentage in exit air) was measured using a mass spectrometer and then its potential impact on dissolved carbon dioxide concentrations qualitatively examined. Greater volumetric air flowrates reduced off-gas carbon dioxide levels more effectively at lower airflow ranges and thus lowered expected dissolved carbon dioxide levels through gas stripping. Lower broth pH values decreased off-gas carbon dioxide levels but increased expected dissolved carbon dioxide levels due to the pH-dependence of the gas/liquid carbon dioxide equilibrium. While back-pressure increases had an insignificant effect on off-gas carbon dioxide levels, they directly affected expected dissolved carbon dioxide levels according to Henry's law. Laboratory studies, conducted using both uninoculated and inoculated fermentation media, quantified the response of the media to pH changes with bicarbonate addition, specifically its buffering capacity. This effect then was related qualitatively to expected dissolved carbon dioxide levels. Higher dissolved carbon dioxide levels, as demonstrated by reduced pH changes with bicarbonate addition, thus would be expected for salt solutions of increased ionic strength and higher protein content media. In addition, pH changes with greater bicarbonate additions declined for fermentation samples taken over the course of a one week cultivation, most likely due to the higher protein content associated with biomass growth. The presence of weak acids/bases initially in the media or formed as metabolic by products, as well as the concentration of buffering ions such as phosphate, also were believed to be important contributing elements to the buffering capacity of the solution. PMID:16232479

  16. Microdilution procedure for antifungal susceptibility testing of Paracoccidioides brasiliensis to amphotericin b and itraconazole

    Directory of Open Access Journals (Sweden)

    E Takahagi-Nakaira

    2009-01-01

    Full Text Available In vitro tests employing microdilution to evaluate fungal susceptibility to antifungal drugs are already standardized for fermentative yeasts. However, studies on the susceptibility of dimorphic fungi such as Paracoccidioides brasiliensis employing this method are scarce. The present work introduced some modifications into antifungal susceptibility testing from the European Committee on Antimicrobial Susceptibility Testing (EUCAST, concerning broth medium and reading time, to determine minimal inhibitory concentration (MIC of amphotericin B and itraconazole against Paracoccidioides brasiliensis. Yeast-like cells of P. brasiliensis (Pb18 strain were tested for susceptibility to amphotericin B and itraconazole in RPMI 1640 medium, supplemented with 2% glucose and nitrogen source and incubated at 35°C. The MIC of amphotericin B and itraconazole against Pb18 were respectively 0.25 µg/mL and 0.002 µg/mL. The results of minimal fungicidal concentration (MFC showed that amphotericin B at 0.25 µg/mL or higher concentrations displayed fungicidal activity against Pb18 while itraconazole at least 0.002 µg/mL has a fungistatic effect on P. brasiliensis. In conclusion, our results showed that the method employed in the present study is reproducible and reliable for testing the susceptibility of P. brasiliensis to antifungal drugs.

  17. Xanthan GumRecovery from Palm Oil-Based Fermentation Broth by Hollow Fibre Microfiltration (MF) Membrane with ProcessOptimisation Using Taguchi Method

    OpenAIRE

    Sufian Soaib, M.; Sabet, M; Krishnan, J.; VPS Veluri, M.

    2013-01-01

    First stage Xanthan recovery (cell and oil separation) from palm oil-based fermentation broth was carried out by hollow fibre microfiltration (MF) using Taguchi method as design of experiment (DOE) to study the effect of four main parameters on Xanthan recovery; transmembrane pressure (TMP), crossflow velocity (CFV), ionic strength (IS) and temperature (T). From S/N ratio larger-the-better analysis, optimum conditions for Xanthan recovery were at level 2 of TMP, IS and T respectively and leve...

  18. Investigation of susceptibility of Staphylococcus species to some antibacterial drugs by disk diffusion and broth microdilution

    OpenAIRE

    Ašanin Jelena; Aksentijević Ksenija; Žutić Milenko; Katić Vera; Krnjaić Dejan; Milić Nenad; Ašanin Ružica; Mišić Dušan

    2012-01-01

    The objective of this work was to identify isolated Staphylococcus species and to investigate their sensitivity to some antibacterial drugs. The material used for these investigations were Staphylococcus isolates originating from milk samples. A total of 25 strains of Staphylococcus isolates were examined, including 24 from milk samples from cows with mastitis, and one strain was isolated from a milk sample from a cow following treatment for mastitis. For p...

  19. Campylobacter growth rates in four different matrices: broiler caecal material, live birds, Bolton broth, and brain heart infusion broth

    OpenAIRE

    Tara Battersby; Desmond Walsh; Paul Whyte; Bolton, Declan J.

    2016-01-01

    Background: The objective of this study was to characterise Campylobacter growth in enrichment broths (Bolton broth, brain heart infusion broth), caecal material (in vitro), and in the naturally infected live broilers (in vivo) in terms of mean lag periods and generation times as well as maximum growth rates and population (cell concentration) achieved. Methods: Bolton and brain heart infusion broths and recovered caecal material were inoculated with 10 poultry strains of Campylobacter (eight...

  20. Campylobacter growth rates in four different matrices: broiler caecal material, live birds, Bolton broth, and brain heart infusion broth

    OpenAIRE

    Battersby, Tara; Walsh, Desmond; Whyte, Paul; Bolton, Declan J.

    2016-01-01

    Background: The objective of this study was to characterise Campylobacter growth in enrichment broths (Bolton broth, brain heart infusion broth), caecal material (in vitro), and in the naturally infected live broilers (in vivo) in terms of mean lag periods and generation times as well as maximum growth rates and population (cell concentration) achieved.Methods: Bolton and brain heart infusion broths and recovered caecal material were inoculated with 10 poultry strains of Campylobacter (eight ...

  1. Development of an Liquid Chromatography-Tandem Mass Spectrometry Method for the Determination of Amoxicillin in Broth Medium and its Application to an In Vitro Pharmacokinetic and Pharmacodynamic Model.

    Science.gov (United States)

    Zhang, Shujing; Yang, Fan; Guo, Beining; Chen, Yuancheng; Wu, Xiaojie; Liang, Wang; Shi, Yaoguo; Zhang, Jing

    2016-02-01

    A simple, rapid and highly sensitive liquid chromatographic-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for the quantification of amoxicillin in broth-a liquid bacterial culture medium. After appropriate dilution with ultrapure water, broth samples containing amoxicillin and an internal standard (IS) were extracted by acetonitrile and dichloromethane. The extract was injected into the system. The analyte and the IS were separated by a prepacked Atlantis C18 column using acetonitrile-0.1% formic acid as a mobile phase and detected by selected reaction monitoring in electrospray ionization positive ion mode. The calibration curve of amoxicillin was linear over the concentration range of 0.05-20.00 µg/mL. The mean recovery of amoxicillin from broth was 71.7%, and the intra- and interday precision and accuracies of the assay were within 10%. Amoxicillin was stable in broth for 12 h at room temperature (24°C), for 6.5 months at -80°C and for 24 h after preparation in an autosampler at room temperature. It has been successfully applied to an in vitro pharmacokinetic (PK) and pharmacodynamic (PD) model in which the broth is used for bacterial growth. The method provides high-throughput biological analysis to facilitate the in vitro PK and PD model of amoxicillin. PMID:26386906

  2. Diagnostic Algorithm Using a Sensitive Broth Culture Method for Detection of Clostridium difficile Toxin from Stool Samples

    Directory of Open Access Journals (Sweden)

    Paul Bayardelle

    2009-01-01

    Full Text Available BACKGROUND: The two-step glutamate dehydrogenase antigencytotoxicity neutralization assay algorithm has been found to be reliable for the diagnosis of toxigenic Clostridium difficile. However, the high sensitivity of the screening method is compromised by the relative low sensitivity of the second step, the direct cytotoxin neutralization assay (DCNA using a fecal filtrate. The objective of the present study was to compare the DCNA with an indirect cytotoxin neutralization assay (ICNA.

  3. Evaluation of antifungal susceptibility testing in Candida isolates by Candifast and disk-diffusion method

    OpenAIRE

    Sidhartha Giri; Anupma Jyoti Kindo

    2014-01-01

    With the increase in invasive fungal infections due to Candida species and resistance to antifungal therapy, in vitro antifungal susceptibility testing is becoming an important part of clinical microbiology laboratories. Along with broth microdilution and disk diffusion method, various commercial methods are being increasingly used for antifungal susceptibility testing, especially in the developed world. In our study, we compared the antifungal susceptibility patterns of 39 isolates of Candid...

  4. Use of the Sensititre Colorimetric Microdilution Panel for Antifungal Susceptibility Testing of Dermatophytes

    OpenAIRE

    Pujol, I.; Capilla, J.; Fernández-Torres, B.; Ortoneda, M.; Guarro, J.

    2002-01-01

    The Sensititre YeastOne antifungal panel was used to test 49 dermatophytes belonging to the species Epidermophyton floccosum, Microsporum gypseum, Microsporum canis, Trichophyton tonsurans, Trichophyton rubrum, and Trichophyton mentagrophytes. The MICs of four antifungals obtained with the Sensititre YeastOne antifungal panel were compared with those obtained by the reference NCCLS microdilution method. The levels of agreement between the two methods (≤2 dilutions) were 81.6% with amphoterici...

  5. Fermented Broth in Tyrosinase- and Melanogenesis Inhibition

    Directory of Open Access Journals (Sweden)

    Chin-Feng Chan

    2014-08-01

    Full Text Available Fermented broth has a long history of applications in the food, pharmaceutical and cosmetic industries. Recently, the use of fermented broth in skin care products is in ascendance. This review investigates the efficacy of fermented broth in inhibiting tyrosinase and melanogenesis. Possible active ingredients and hypopigmentation mechanisms of fermented broth are discussed, and potential applications of fermented broth in the cosmetic industry are also addressed.

  6. Evaluation of the Etest Method for Determining Fluconazole Susceptibilities of 402 Clinical Yeast Isolates by Using Three Different Agar Media

    OpenAIRE

    Pfaller, M A; Messer, S. A.; Karlsson, Å.; Bolmström, A.

    1998-01-01

    The performance of the Etest for fluconazole susceptibility testing of 402 yeast isolates was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35°C. Etest MICs were determined with RPMI agar containing 2% glucose (RPG), Casitone agar (CAS), and Mueller-Hinton agar (MHA) and were read after incubation for 48 h at 35°C. The yeast isolates...

  7. Evaluation of Etest Method for Determining Caspofungin (MK-0991) Susceptibilities of 726 Clinical Isolates of Candida Species

    OpenAIRE

    Pfaller, M A; Messer, S. A.; Mills, K.; Bolmström, A.; Jones, R N

    2001-01-01

    The performance of the Etest for testing the susceptibilities to caspofungin (MK-0991) of 726 isolates of Candida spp. was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35°C. MICs were determined by Etest for all 726 isolates with RPMI agar containing 2% glucose (RPG) and were read after incubation for 48 h at 35°C. The Candida isola...

  8. Ultra scale-down characterization of the impact of conditioning methods for harvested cell broths on clarification by continuous centrifugation-Recovery of domain antibodies from rec E. coli.

    Science.gov (United States)

    Chatel, Alex; Kumpalume, Peter; Hoare, Mike

    2014-05-01

    The processing of harvested E. coli cell broths is examined where the expressed protein product has been released into the extracellular space. Pre-treatment methods such as freeze-thaw, flocculation, and homogenization are studied. The resultant suspensions are characterized in terms of the particle size distribution, sensitivity to shear stress, rheology and solids volume fraction, and, using ultra scale-down methods, the predicted ability to clarify the material using industrial scale continuous flow centrifugation. A key finding was the potential of flocculation methods both to aid the recovery of the particles and to cause the selective precipitation of soluble contaminants. While the flocculated material is severely affected by process shear stress, the impact on the very fine end of the size distribution is relatively minor and hence the predicted performance was only diminished to a small extent, for example, from 99.9% to 99.7% clarification compared with 95% for autolysate and 65% for homogenate at equivalent centrifugation conditions. The lumped properties as represented by ultra scale-down centrifugation results were correlated with the basic properties affecting sedimentation including particle size distribution, suspension viscosity, and solids volume fraction. Grade efficiency relationships were used to allow for the particle and flow dynamics affecting capture in the centrifuge. The size distribution below a critical diameter dependent on the broth pre-treatment type was shown to be the main determining factor affecting the clarification achieved. PMID:24284936

  9. Comparison of different phenotypic and genotypic methods for the detection of methicillin-resistant Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Abbas Farahani

    2013-01-01

    Full Text Available Background: Staphylococcus aureus is known as a powerful pathogen that causes various infections. Emergence of methicillin-resistant S. aureus (MSRA is responsible for nosocomial and community-acquired infections worldwide. Aims: The present study aimed to evaluate the performance and ability of eight different phenotypic and genotypic methods for the detection of MSRA. Materials and Methods: A total of 186 S. aureus isolates were defined as methicillin-susceptible S. aureus (MSSA; 95 and MSRA (91 using polymerase chain reaction (PCR as the gold standard. Susceptibility to methicillin was investigated using oxacillin, methicillin, cefotetan, cefoxitin, and cefmetazole disks, by oxacillin Adata Tab and strips. For all S. aureus isolates minimal inhibitory concentrations of oxacillin were determined using the broth microdilution method according to Clinical and Laboratory Standards Institute guidelines. Results: Among the diagnostic methods studied, broth microdilution and the cefoxitin disk had the highest specificity (98.9 and 94.7%, sensitivity (100 and 98.9%, and concordance with PCR results (98.9 and 93.6%. The cefotetan and cefmetazole disks had the lowest concordance with PCR results. Conclusion: Our results suggest that microdilution and cefoxitin disk methods have high sensitivities compared with other methods for detection of MSRA. The cefoxitin disk method may be preferred in clinical laboratories because it is easy to perform and does not require special equipment.

  10. Separation technologies for the recovery and dehydration of alcohols from fermentation broths

    Science.gov (United States)

    Multi-column distillation followed by molecular sieve adsorption is currently the standard method for producing fuel grade ethanol from dilute fermentation broths in modern corn-to-ethnol facilities. As the liquid biofuels industry transitions to lignocellulosic feedstocks, expan...

  11. βlasEN: Microdilution Panel for Identifying β-Lactamases Present in Isolates of Enterobacteriaceae

    OpenAIRE

    Sanders, Christine C.; Ehrhardt, Anton F.; Moland, Ellen Smith; Kenneth S. Thomson; Zimmer, Barbara; Roe, Darcie E.

    2002-01-01

    A dried investigational use-only microdilution panel named βlasEN (a short named derived from the panel’s purpose, to identify β-lactamases in Enterobacteriaceae) containing 10 β-lactam drugs with and without β-lactamase inhibitors was developed to identify β-lactamases among clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter koseri, Citrobacter freundii group, Enterobacter spp., and Serratia marcescens. The MICs obtained with a collection of 383 org...

  12. The risk of lead contamination in bone broth diets.

    Science.gov (United States)

    Monro, J A; Leon, R; Puri, B K

    2013-04-01

    The preparation and consumption of bone broth is being increasingly recommended to patients, for example as part of the gut and psychology syndrome (GAPS) diet for autism, attention-deficit hyperactivity disorder, dyslexia, dyspraxia, depression and schizophrenia, and as part of the paleolithic diet. However, bones are known to sequester the heavy metal lead, contamination with which is widespread throughout the modern environment. Such sequestered lead can then be mobilised from the bones. We therefore hypothesised that bone broth might carry a risk of being contaminated with lead. A small, blinded, controlled study of lead concentrations in three different types of organic chicken broth showed that such broths do indeed contain several times the lead concentration of the water with which the broth is made. In particular, broth made from skin and cartilage taken off the bone once the chicken had been cooked with the bones in situ, and chicken-bone broth, were both found to have markedly high lead concentrations, of 9.5 and 7.01 μg L(-1), respectively (compared with a control value for tap water treated in the same way of 0.89 μg L(-1)). In view of the dangers of lead consumption to the human body, we recommend that doctors and nutritionists take the risk of lead contamination into consideration when advising patients about bone broth diets. PMID:23375414

  13. Filtration Process of the Spiramycin Fermentation Broth

    Institute of Scientific and Technical Information of China (English)

    1999-01-01

    This paper presents the effects of acidity and additive pretreatment on the filtering rate and Spiramycin (SPM) concentration in the filter liquor of SPM fermentation broth. The experimental results show that the SPM peak value in filter liquor is obtained at pH 5.5 with either 0.1% methanal or 0.1% BAPE. It is also indicated that there exists a dissolution equilibrium of proteins from the experiment results. The soluble proteins are denatured due to the too high/low acidity and then precipitate. Usually, the amount of soluble proteins reaches its lowest level in pH range of 6.0-6.5. The protein precipitation will, together with other suspended solids particles, contribute to the final SPM concentration in the filter liquor. This paper assumes that the contribution is the result of the adsorption equilibrium of SPM on the surfaces of suspended solids. For a satisfactory explanation, the revised Langmuir adsorption theory was employed and a model was developed.

  14. Automated HPLC monitoring of broth components on bioreactors

    OpenAIRE

    Favre, Eric; Pugeaud, Patrick; Raboud, Jean Philippe; Péringer, Paul

    1989-01-01

    Under proper operating conditions, a low dead volume continuous filtration module operated on biological broths (yeast and bacteria suspensions in stirred reactors) still fulfills the flow-rate requirements of an analytical apparatus (for example HPLC or FIA) without membrane regeneration. The filtrate stream has been successfully connected to a bioreactor in order to perform the automated HPLC analysis of broth components. The monitoring of the carbon source (lactose), and minor products (gl...

  15. Early and Efficient Detection of Mycobacterium tuberculosis in Sputum by Microscopic Observation of Broth Cultures

    OpenAIRE

    Benson R Kidenya; Rodrick Kabangila; Peck, Robert N; Mshana, Stephen E; Webster, Lauren E.; Koenig, Serena P; Warren D Johnson; Fitzgerald, Daniel W.

    2013-01-01

    Early, efficient and inexpensive methods for the detection of pulmonary tuberculosis are urgently needed for effective patient management as well as to interrupt transmission. These methods to detect M. tuberculosis in a timely and affordable way are not yet widely available in resource-limited settings. In a developing-country setting, we prospectively evaluated two methods for culturing and detecting M. tuberculosis in sputum. Sputum samples were cultured in liquid assay (micro broth cultur...

  16. Cross-flow membrane microfiltration of a bacteriol fermentation broth.

    Science.gov (United States)

    Nagata, N; Herouvis, K J; Dziewulski, D M; Belfort, G

    1989-08-01

    Although cross-flow membrane filtration is a very attractive option for harvesting cells and recovering enzymes from cell homogenates, the process is not without its problems. Foremost of these is the deposit of dissolved and suspended solutes onto the membrane surface during operation. The formation of these dense and sometimes compressive sublayers (often called cakes) offers additional resistance to axial and permeate flows and often affects the retention characteristics of the process. In view of the complex nature of the sublayer formation process and its sensitivity to cross-flow velocity, this investigation was undertaken to determine the main factors responsible for the decline in performance during the harvesting of B. polymyxa broth by membrane microfiltration. System parameters varied include axial flow rate, concentration of cells, proteins and other components in the feed, membrane materials (ceramic, polypropylene, and stainless steel), and cleaning methods. To help explain the observed results, a new mass transport model-the solids flux model-based on the assumptions that back migration of particles from the sublayer or membrane surface is negligible and that particles that reach the solid-solution interface attach (stick) completely, is tested. Using a variety of diagnostic methods, magnesium ammonium phosphate precipitate is formed during steam sterilization of the medium and is implicated as the major foulant in this study. PMID:18588126

  17. Spherical UO2 Size and Microstructure on Broth Preparation Conditions

    International Nuclear Information System (INIS)

    Without a doubt, the High Temperature Reactor will play a dominant role in the worldwide fleet of nuclear reactors of the next decade. It is being conducted by many countries, such as US, Japan, China, and so on, and mainly promoted for electricity production and high temperature heat. There are two general designs for He gas cooled reactor; types of the modular high temperature gas cooled reactor and the pebble bed reactor. Both reactor types use a basic fuel concept based on the dispersion of TRISO coated particles in graphite. The TRISO coated particle for these purposes prepared by pyro-carbon and silicone carbide coatings on a spherical UO2 kernel surface as a fissile material. Generally, UO2 kernels are produced by using the modified sol-gel process, wet process, known as the GSP(gel supported precipitation) method. This chemical route was well-known to the potential kernel fabrication processes. Figure 1 depicts a flow diagram of the VTHR fuel preparation process. After the formation of broth solution contained UN(Uranyl Nitrate) solution, PVA solution, and organic additives, spherical droplets are formed by a vibrating nozzle system. Spherical droplets are converted to liquid-ADU spheres in gelation column, and then finally aged-ADU gel particles in NH4OH solution. Following the drying and calcining, reduction of the calcined kernels to UO2 was carried out. This reduced UO2 particles were converted to a dense structure UO2 through the sintering process

  18. Binding of Todd-Hewitt broth antigens by Streptococcus mutans.

    Science.gov (United States)

    Stinson, M W; Jones, C A

    1983-01-01

    Streptococcus mutans 10449, grown in chemically defined culture medium, was tested for its ability to bind 3H-labeled Todd-Hewitt broth components (greater than 12,000 Mr). Maximum adsorption of radioactivity occurred within 5 min at room temperature, and cell-bound material was not completely removed by extended washing with buffer. Heat-killed, arsenate-inhibited, and viable bacteria bound similar quantities. Only 0.09% of the radioactivity in the preparation of high Mr Todd-Hewitt broth components was removed by absorption with excess numbers of S. mutans 10449 cells. Binding followed saturation kinetics and was competitively inhibited by unlabeled medium components, both the dialyzable and nondialyzable fractions. Other oral streptococci were also found to bind these complex medium components. Rabbit antiserum elicited to the high-molecular-weight Todd-Hewitt broth components reacted with monkey cardiac muscle and with S. mutans coated with medium components. Absorption of the anti-Todd-Hewitt broth serum with homogenized heart removed antibodies that reacted with Todd-Hewitt broth-coated S. mutans. Therefore, the tissue-specific antigens of this beef heart infusion medium that adsorb to S. mutans can interfere with the detection and characterization of antigens shared by these bacteria and animal tissues. Images PMID:6852915

  19. Flow cytometry susceptibility testing for conventional antifungal drugs and Comparison with the NCCLS Broth Macrodilution Test

    Directory of Open Access Journals (Sweden)

    M.J. Najafzadeh

    2009-08-01

    Full Text Available Introduction: During the last decade, the incidence of fungal infection has been increased in many countries. Because of the advent of resistant to antifungal agents, determination of an efficient strategic plan for treatment of fungal disease is an important issue in clinical mycology. Many methods have been introduced and developed for determination of invitro susceptibility tests. During the recent years, flow cytometry has developed to solving the problem and many papers have documented the usefulness of this technique. Materials and methods: As the first step, the invitro susceptibility of standard PTCC (Persian Type of Culture Collection strain and some clinical isolates of Candida consisting of Candida albicans, C. dubliniensis, C. glabrata, C. kefyer and C. parapsilosis were evaluated by macrodilution broth method according to NCCLS (National Committee for Clinical Laboratory Standards guidelines and flow cytometry susceptibility test. Results:  The data indicated that macro dilution broth methods and flow cytometry have the same results in determination of MIC (Minimum Inhibitory Concentration for amphotericin B, clotrimazole, fluconazole, ketoconazole and miconazole in C. albicans PTCC 5027 as well as clinical Candida isolates, such as C.albicans, C.dubliniensis, C.glabrata C.kefyr, and C.parapsilosis. Discussion: Comparing the results obtained by macrodilution broth and flow cytometry methods revealed that flow cytometry was faster. It is suggested that flow cytometry susceptibility test can be used as a powerful tool for determination of MIC and administration of the best antifungal drug in treatment of patients with Candida infections.

  20. Early and efficient detection of Mycobacterium tuberculosis in sputum by microscopic observation of broth cultures.

    Directory of Open Access Journals (Sweden)

    Benson R Kidenya

    Full Text Available Early, efficient and inexpensive methods for the detection of pulmonary tuberculosis are urgently needed for effective patient management as well as to interrupt transmission. These methods to detect M. tuberculosis in a timely and affordable way are not yet widely available in resource-limited settings. In a developing-country setting, we prospectively evaluated two methods for culturing and detecting M. tuberculosis in sputum. Sputum samples were cultured in liquid assay (micro broth culture in microplate wells and growth was detected by microscopic observation, or in Löwenstein-Jensen (LJ solid media where growth was detected by visual inspection for colonies. Sputum samples were collected from 321 tuberculosis (TB suspects attending Bugando Medical Centre, in Mwanza, Tanzania, and were cultured in parallel. Pulmonary tuberculosis cases were diagnosed using the American Thoracic Society diagnostic standards. There were a total of 200 (62.3% pulmonary tuberculosis cases. Liquid assay with microscopic detection detected a significantly higher proportion of cases than LJ solid culture: 89.0% (95% confidence interval [CI], 84.7% to 93.3% versus 77.0% (95% CI, 71.2% to 82.8% (p = 0.0007. The median turn around time to diagnose tuberculosis was significantly shorter for micro broth culture than for the LJ solid culture, 9 days (interquartile range [IQR] 7-13, versus 21 days (IQR 14-28 (p<0.0001. The cost for micro broth culture (labor inclusive in our study was US $4.56 per sample, versus US $11.35 per sample for the LJ solid culture. The liquid assay (micro broth culture is an early, feasible, and inexpensive method for detection of pulmonary tuberculosis in resource limited settings.

  1. Modeling of mixing in stirred bioreactors 4. mixing time for aerated bacteria, yeasts and fungus broths

    Directory of Open Access Journals (Sweden)

    Cascaval Dan

    2004-01-01

    Full Text Available The mixing time for bioreactors depends mainly on the rheoiogicai properties of the broths, the biomass concentration and morphology, mixing system characteristics and fermentation conditions. For quantifying the influence of these factors on the mixing efficiency for stirred bioreactors, aerated broths of bacteria (P. shermanii, yeasts (S. cerevisiae and fungi (P. chrysogenum, free mycelia and mycelial aggregates of different concentrations have been investigated using a laboratory bioreactor with a double turbine impeller. The experimental data indicated that the influence of the rotation speed, aeration rate and stirrer positions on the mixing intensity strongly differ from one system to another and must be correlated with the microorganism characteristics, namely: the biomass concentration and morphology. Moreover, compared with non-aerated broths, variations of the mixing time with the considered parameters are very different, due to the complex flow mechanism of gas-liquid dispersions. By means of the experimental data and using a multiregression analysis method some mathematical correlations for the mixing time of the general form: tm = a1*Cx2+a2*Cx+a3*IgVa+a4-N2+a5-N+a6/a7*L2+a8*L+a9 were established. The proposed equations offer good agreement with the experiments, the average deviation being ±6.7% - ±9.4 and are adequate for the flow regime Re < 25,000.

  2. Green synthesis of gold nanoparticles using Cinnamomum zeylanicum leaf broth

    Science.gov (United States)

    Smitha, S. L.; Philip, Daizy; Gopchandran, K. G.

    2009-10-01

    Development of biologically inspired experimental processes for the synthesis of nanoparticles is an important branch of nanotechnology. The synthesis of gold nanoparticles using Cinnamomum zeylanicum leaf broth as the reducing agent is reported. The morphology of the particles formed consists of a mixture of gold nanoprisms and spheres with fcc (1 1 1) structure of gold. At lower concentrations of the extract, formation of prism shaped Au particles dominates, while at higher concentrations almost spherical particles alone are observed. Good crystallinity of the nanoparticles with fcc phase is evident from XRD patterns, clear lattice fringes in the high resolution TEM image and bright circular rings in the SAED pattern. Au nanoparticles grown are observed to be photoluminescent and the intensity of photoemission is found to increase with increase in leaf broth concentration. The ability to modulate the shape of nanoparticles as observed in this study for gold nanoparticles opens up the exciting possibility of developing further synthetic routes employing ecofriendly sources.

  3. Carbon black selection from simulated broth solution for ADU gel spheres

    International Nuclear Information System (INIS)

    The VHTR (Very High Temperature Gas Reactor) is one of the reactor concepts in the Gen IV International Collaboration. The nuclear fuel of a VHTR in the US is based on microspheres containing a mixture of UO2 and UC2 coated with multi carbon layers and a SiC layer. This mixture is called a 'UCO (uranium oxi carbide)' kernel. The fabrication process of this kernel was based on the sol-gel method between an ADUN and HMTA and urea, a process referred to as internal gelation. UCO kernel microspheres were first prepared at ORNL in the late 1970s. CB(Carbon Black) as a carbon source in the final UCO kernel is added during the broth solution preparation, in the processing of UCO kernel fabrication. The preparation of a good quality UCO kernel is very difficult due to the homogeneous distribution of carbon in a UCO kernel. The key requirement to obtain a good quality kernel is a uniform distribution of carbon in the ADU gel sphere forming process before the thermal treatment, i.e., during the gel formation step. The internal gelation concept was adapted in ADU gel sphere fabrication in the ORNL process of the US. Generally, UO2 kernel microspheres are prepared by an internal gelation method (USA, India) or external gelation method (Germany, China, Japan). The UCO kernel microspheres prepared only in the US, use an internal gelation method. A material flow chart on the preparation of the microsphere kernel is simply shown in Fig. 1. The broth solution preparation, the raw material, additives, and thermal steps such as calcining and sintering processes were different to compared with the external gelation and internal gelation methods. In this study, we first carried out the matching CB selection experiments among the various kinds of CBs in a broth solution, for UCO kernel preparation using an external gelation method.

  4. Evaluation of Fastidious Anaerobe Broth as a blood culture medium.

    OpenAIRE

    Ganguli, L. A.; Turton, L J; Tillotson, G S

    1982-01-01

    Three commercial blood culture media were compared with a freshly prepared cooked meat medium in tests to stimulate the recovery of small inocula of anaerobic and aerobic bacteria in routine blood cultures. The cooked meat medium gave the most reliable recovery and supported continued viability, whilst Fastidious Anaerobe Broth (LAB M) was a good alternative. Results with Southern Group thioglycollate and Difco Thiol were less satisfactory as delays in recovery and loss of viability occurred ...

  5. Mutagenicity potential of commercial broth cubes at varying concentrations

    International Nuclear Information System (INIS)

    Today, there has been a growing concern on the mutagenicity potential of environmental chemical systems. These environmental chemicals such as pesticides, food additives, synthetic drugs, water and atmospheric pollutants are possible causes of mutagenic activity. Meat products and some meat flavorings, were also reported to exhibit mutagenic activity. And since these products are normal part of the daily human diet, there is a need for extensive studies regarding the possible mutagenic activity associated with these products. This study aimed to evaluate the mutagenicity potential of commercial broth cubes at varying concentration. The researchers sought to answer the following questions: 1. Do beef, pork and chicken broth cubes exhibit mutagenic activity? 2. Are there significant differences in the mutagenic activity among the three samples? 3. Are these significant differences in the mutagenic activity exhibited by each of the samples compared to that of Mitomycin-C (positive control)? 4. Which of the sample of each specific concentration exhibit the greatest mutagenic activity? Three specific concentrations of beef, pork and chicken broth cubes were prepared and their mutagenicity potential was evaluated by using the Micronucleus test. The formation of micro nucleated polychromatic and micro nucleated normo chromatic erythrocytes in bone marrow cells of mice treated with these samples were detected using a Carl-Zeiss photo microscope. The statistical tool used to test the validity of the null hypothesis was analysis of variance using randomized complete block design and independent T- test. (author)

  6. Background and Objective: Listeria monocytogenes is a bacterium transferred by foods and is the agent of many sporadic and epidemic diseases in humans. This study aimed to investigate the prevalence of L. monocytogenes and to determinine their antibiotic resistance profile in red meats. Material and Methods: this cross-sectional study was performed on 400 red meat samples obtained from industrial slaughterhouses placed in Kerman, Iran. First, the samples were enriched with Simultaneous Enrichment Broth (SEB, and then plated onto Palcam agar and Tryptic Soy Broth Yeast Extract Broth (TSAYE. After identification of the isolates based on biochemical tests and PCR, the isolates were checked for their antibiotic resistance profile using disk Diffusion Results: of 400 samples, 12 samples (3% were contaminated with different species of Listeria. Using PCR, hly gene was recognized in eight samples (2% of L. monocytogenes. Furthermore, there was a significant difference in isolation rate of lamb samples compared to cow ones. While all of the isolates were resistant to clindamycin, amikacin and chloramphenicol, they were sensitive to penicillin. Conclusion: in spite of low rate of infection in red meat samples in Kerman city, due to high risk of Listeria contamination in red meats, we recommend applying a routine screening to identify this bacterium in our county.

    Directory of Open Access Journals (Sweden)

    Kargar, M. (PhD

    2014-11-01

    Full Text Available Background and Objective: Listeria monocytogenes is a bacterium transferred by foods and is the agent of many sporadic and epidemic diseases in humans. This study aimed to investigate the prevalence of L. monocytogenes and to determinine their antibiotic resistance profile in red meats. Material and Methods: this cross-sectional study was performed on 400 red meat samples obtained from industrial slaughterhouses placed in Kerman, Iran. First, the samples were enriched with Simultaneous Enrichment Broth (SEB, and then plated onto Palcam agar and Tryptic Soy Broth Yeast Extract Broth (TSAYE. After identification of the isolates based on biochemical tests and PCR, the isolates were checked for their antibiotic resistance profile using disk Diffusion Results: of 400 samples, 12 samples (3% were contaminated with different species of Listeria. Using PCR, hly gene was recognized in eight samples (2% of L. monocytogenes. Furthermore, there was a significant difference in isolation rate of lamb samples compared to cow ones. While all of the isolates were resistant to clindamycin, amikacin and chloramphenicol, they were sensitive to penicillin. Conclusion: in spite of low rate of infection in red meat samples in Kerman city, due to high risk of Listeria contamination in red meats, we recommend applying a routine screening to identify this bacterium in our county

  7. BetalasEN: microdilution panel for identifying beta-lactamases present in isolates of Enterobacteriaceae.

    Science.gov (United States)

    Sanders, Christine C; Ehrhardt, Anton F; Moland, Ellen Smith; Thomson, Kenneth S; Zimmer, Barbara; Roe, Darcie E

    2002-01-01

    A dried investigational use-only microdilution panel named betalasEN (a short named derived from the panel's purpose, to identify beta-lactamases in Enterobacteriaceae) containing 10 beta-lactam drugs with and without beta-lactamase inhibitors was developed to identify beta-lactamases among clinical isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Citrobacter koseri, Citrobacter freundii group, Enterobacter spp., and Serratia marcescens. The MICs obtained with a collection of 383 organisms containing well-characterized beta-lactamases were used to develop numeric codes and logic pathways for computerized analysis of results. The resultant logic pathways and betalasEN panel were then used to test and identify beta-lactamases among 885 isolates of Enterobacteriaceae recovered in cultures obtained at six different hospital laboratories across the United States. beta-Lactamases present in 801 (90.5%) of the 885 isolates were identified by betalasEN by using the existing logic pathways and codes or after minor modifications were made to the existing codes. The 84 strains that gave codes that betalasEN could not identify were collected, reidentified, and retested by using betalasEN. Three strains had been misidentified, 54 strains gave different codes upon repeat testing that could be identified by betalasEN, and 27 strains repeated new codes. The beta-lactamases in these strains were identified, and the new codes were added to the betalasEN logic pathways. These results indicate that betalasEN can identify clinically important beta-lactamases among most isolates of Enterobacteriaceae. The results also show that good quality control and attention to proper performance of the tests are essential to the correct performance of betalasEN. PMID:11773104

  8. Broth medium for enrichment of Vibrio fluvialis from the environment.

    OpenAIRE

    Nishibuchi, M; Roberts, N C; Bradford, H B; Seidler, R J

    1983-01-01

    A medium was designed for the enrichment and enumeration of Vibrio fluvialis from environmental samples. The medium contains 1% peptone plus 4% sodium chloride and 5 micrograms of novobiocin per ml, pH 8.5. This V. fluvialis enrichment medium (FEM) was tested, in comparison with alkaline peptone (AP), in field samplings. A total of 177 samples (estuarine waters and sediment, sewage, and crabs) collected over a 14-month period were examined with FEM and with AP broth. Results showed that FEM w...

  9. In vitro activities of clinafloxacin against contemporary clinical bacterial isolates from 10 North American centers.

    Science.gov (United States)

    Fuchs, P C; Barry, A L; Brown, S D

    1998-05-01

    Clinafloxacin was more active than ciprofloxacin against 4,213 aerobic and facultative anaerobic bacterial isolates from 10 medical centers, as tested by broth microdilution and disk diffusion methods. The percentage of 201 anaerobes susceptible to clinafloxacin by broth microdilution was comparable to cefoxitin. Our data support the proposed disk diffusion interpretive criteria for aerobic bacteria with 5-microg clinafloxacin disks. PMID:9593166

  10. Comparative evaluation of antibacterial activity of caffeic acid phenethyl ester and PLGA nanoparticle formulation by different methods

    Science.gov (United States)

    Arasoglu, Tülin; Derman, Serap; Mansuroglu, Banu

    2016-01-01

    The aim of the present study was to evaluate the antimicrobial activity of nanoparticle and free formulations of the CAPE compound using different methods and comparing the results in the literature for the first time. In parallel with this purpose, encapsulation of CAPE with the PLGA nanoparticle system (CAPE-PLGA-NPs) and characterization of nanoparticles were carried out. Afterwards, antimicrobial activity of free CAPE and CAPE-PLGA-NPs was determined using agar well diffusion, disk diffusion, broth microdilution and reduction percentage methods. P. aeroginosa, E. coli, S. aureus and methicillin-resistant S. aureus (MRSA) were chosen as model bacteria since they have different cell wall structures. CAPE-PLGA-NPs within the range of 214.0 ± 8.80 nm particle size and with an encapsulation efficiency of 91.59 ± 4.97% were prepared using the oil-in-water (o-w) single-emulsion solvent evaporation method. The microbiological results indicated that free CAPE did not have any antimicrobial activity in any of the applied methods whereas CAPE-PLGA-NPs had significant antimicrobial activity in both broth dilution and reduction percentage methods. CAPE-PLGA-NPs showed moderate antimicrobial activity against S. aureus and MRSA strains particularly in hourly measurements at 30.63 and 61.25 μg ml-1 concentrations (both p 0.05). In the reduction percentage method, in which the highest results of antimicrobial activity were obtained, it was observed that the antimicrobial effect on S. aureus was more long-standing (3 days) and higher in reduction percentage (over 90%). The appearance of antibacterial activity of CAPE-PLGA-NPs may be related to higher penetration into cells due to low solubility of free CAPE in the aqueous medium. Additionally, the biocompatible and biodegradable PLGA nanoparticles could be an alternative to solvents such as ethanol, methanol or DMSO. Consequently, obtained results show that the method of selection is extremely important and will influence the

  11. Studies of polypropylene membrane fouling during microfiltration of broth with Citrobacter freundii bacteria

    Directory of Open Access Journals (Sweden)

    Gryta Marek

    2015-12-01

    Full Text Available In this work a fouling study of polypropylene membranes used for microfiltration of glycerol solutions fermented by Citrobacter freundii bacteria was presented. The permeate free of C. freundii bacteria and having a turbidity in the range of 0.72–1.46 NTU was obtained. However, the initial permeate flux (100–110 L/m2h at 30 kPa of transmembrane pressure was decreased 3–5 fold during 2–3 h of process duration. The performed scanning electron microscope observations confirmed that the filtered bacteria and suspensions present in the broth formed a cake layer on the membrane surface. A method of periodical module rinsing was used for restriction of the fouling influence on a flux decline. Rinsing with water removed most of the bacteria from the membrane surface, but did not permit to restore the initial permeate flux. It was confirmed that the irreversible fouling was dominated during broth filtration. The formed deposit was removed using a 1 wt% solution of sodium hydroxide as a rinsing solution.

  12. Effect of fermented broth from lactic acid bacteria on pathogenic bacteria proliferation.

    Science.gov (United States)

    Gutiérrez, S; Martínez-Blanco, H; Rodríguez-Aparicio, L B; Ferrero, M A

    2016-04-01

    In this study, the effect that 5 fermented broths of lactic acid bacteria (LAB) strains have on the viability or proliferation and adhesion of 7 potentially pathogenic microorganisms was tested. The fermented broth from Lactococcus lactis C660 had a growth inhibitory effect on Escherichia coli K92 that reached of 31%, 19% to Pseudomonas fluorescens, and 76% to Staphylococcus epidermidis. The growth of Staph. epidermidis was negatively affected to 90% by Lc. lactis 11454 broth, whereas the growth of P. fluorescens (25%) and both species of Staphylococcus (35% to Staphylococcus aureus and 76% to Staph. epidermidis) were inhibited when they were incubated in the presence of Lactobacillus casei 393 broth. Finally, the fermented broth of Lactobacillus rhamnosus showed an inhibitory effect on growth of E. coli K92, Listeria innocua, and Staph. epidermidis reached values of 12, 28, and 76%, respectively. Staphylococcus epidermidis was the most affected strain because the effect was detected from the early stages of growth and it was completely abolished. The results of bacterial adhesion revealed that broths from Lc. lactis strains, Lactobacillus paracasei, and Lb. rhamnosus caused a loss of E. coli K92 adhesion. Bacillus cereus showed a decreased of adhesion in the presence of the broths of Lc. lactis strains and Lb. paracasei. Listeria innocua adhesion inhibition was observed in the presence of Lb. paracasei broth, and the greatest inhibitory effect was registered when this pathogenic bacterium was incubated in presence of Lc. lactis 11454 broth. With respect to the 2 Pseudomonas, we observed a slight adhesion inhibition showed by Lactobacillus rhamnosus broth against Pseudomonas putida. These results confirm that the effect caused by the different LAB assayed is also broth- and species-specific and reveal that the broth from LAB tested can be used as functional bioactive compounds to regulate the adhesion and biofilm synthesis and ultimately lead to preventing food and

  13. Recovery of Xanthan Gum from Palm Oil-Based Fermentation Broth by Diafiltration with Flat Polysulfone Microfiltration (MF) Membrane

    OpenAIRE

    Sufian So’aib, M.; Krishnan, J.; Veluri, M. V. P. S.

    2014-01-01

    Xanthan gum recovery from palm oil-based broth by diafiltration was carried out using flat microfiltration (MF) membrane. Optimization of process parameters such as transmembrane pressure (TMP), crossflow velocity (CFV), ionic strength (IS) and diafiltration factor (DF) was performed by Taguchi method using signal-to-noise (S/N) ratio of larger-the-better criterion yielding the following optimum conditions: level 1, level 2, level 3, and level 2, respectively, corresponding to Xanthan recover...

  14. An integrated platform for gas-diffusion separation and electrochemical determination of ethanol on fermentation broths

    International Nuclear Information System (INIS)

    Highlights: • Integrated platform was developed to determine ethanol in fermentation broths. • The designed system integrates gas diffusion separation with voltammetric detection. • Detector relied on Ni(OH)2-modified electrode stabilized by Co2+ and Cd2+ insertion. • Separation was made by PTFE membrane separating sample from electrolyte (receptor). • Despite the sample complexity, accurate tests were achieved by direct interpolation. - Abstract: An integrated platform was developed for point-of-use determination of ethanol in sugar cane fermentation broths. Such analysis is important because ethanol reduces its fuel production efficiency by altering the alcoholic fermentation step when in excess. The custom-designed platform integrates gas diffusion separation with voltammetric detection in a single analysis module. The detector relied on a Ni(OH)2-modified electrode. It was stabilized by uniformly depositing cobalt and cadmium hydroxides as shown by XPS measurements. Such tests were in accordance with the hypothesis related to stabilization of the Ni(OH)2 structure by insertion of Co2+ and Cd2+ ions in this structure. The separation step, in turn, was based on a hydrophobic PTFE membrane, which separates the sample from receptor solution (electrolyte) where the electrodes were placed. Parameters of limit of detection and analytical sensitivity were estimated to be 0.2% v/v and 2.90 μA % (v/v)−1, respectively. Samples of fermentation broth were analyzed by both standard addition method and direct interpolation in saline medium based-analytical curve. In this case, the saline solution exhibited ionic strength similar to those of the samples intended to surpass the tonometry colligative effect of the samples over analyte concentration data by attributing the reduction in quantity of diffused ethanol vapor majorly to the electrolyte. The approach of analytical curve provided rapid, simple and accurate analysis, thus contributing for deployment of point

  15. An integrated platform for gas-diffusion separation and electrochemical determination of ethanol on fermentation broths

    Energy Technology Data Exchange (ETDEWEB)

    Giordano, Gabriela Furlan [Microfabrication Laboratory, Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP 13083-970 (Brazil); Department of Analytical Chemistry, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil); National Institute of Science and Technology of Bioanalytics, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil); Vieira, Luis Carlos Silveira; Gobbi, Angelo Luiz [Microfabrication Laboratory, Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP 13083-970 (Brazil); Lima, Renato Sousa [Microfabrication Laboratory, Brazilian Nanotechnology National Laboratory (LNNano), Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, SP 13083-970 (Brazil); Department of Analytical Chemistry, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil); National Institute of Science and Technology of Bioanalytics, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil); Kubota, Lauro Tatsuo, E-mail: kubota@iqm.unicamp.br [Department of Analytical Chemistry, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil); National Institute of Science and Technology of Bioanalytics, Institute of Chemistry – UNICAMP, Campinas, SP 13083-970 (Brazil)

    2015-05-22

    Highlights: • Integrated platform was developed to determine ethanol in fermentation broths. • The designed system integrates gas diffusion separation with voltammetric detection. • Detector relied on Ni(OH){sub 2}-modified electrode stabilized by Co{sup 2+} and Cd{sup 2+} insertion. • Separation was made by PTFE membrane separating sample from electrolyte (receptor). • Despite the sample complexity, accurate tests were achieved by direct interpolation. - Abstract: An integrated platform was developed for point-of-use determination of ethanol in sugar cane fermentation broths. Such analysis is important because ethanol reduces its fuel production efficiency by altering the alcoholic fermentation step when in excess. The custom-designed platform integrates gas diffusion separation with voltammetric detection in a single analysis module. The detector relied on a Ni(OH){sub 2}-modified electrode. It was stabilized by uniformly depositing cobalt and cadmium hydroxides as shown by XPS measurements. Such tests were in accordance with the hypothesis related to stabilization of the Ni(OH){sub 2} structure by insertion of Co{sup 2+} and Cd{sup 2+} ions in this structure. The separation step, in turn, was based on a hydrophobic PTFE membrane, which separates the sample from receptor solution (electrolyte) where the electrodes were placed. Parameters of limit of detection and analytical sensitivity were estimated to be 0.2% v/v and 2.90 μA % (v/v){sup −1}, respectively. Samples of fermentation broth were analyzed by both standard addition method and direct interpolation in saline medium based-analytical curve. In this case, the saline solution exhibited ionic strength similar to those of the samples intended to surpass the tonometry colligative effect of the samples over analyte concentration data by attributing the reduction in quantity of diffused ethanol vapor majorly to the electrolyte. The approach of analytical curve provided rapid, simple and accurate

  16. Antimicrobial activity of cefmetazole (CS-1170) and recommendations for susceptibility testing by disk diffusion, dilution, and anaerobic methods.

    Science.gov (United States)

    Jones, R N; Barry, A L; Fuchs, P C; Thornsberry, C

    1986-12-01

    Cefmetazole, formerly CS-1170, was found to have antimicrobial activity slightly superior to that of cefoxitin but a clinically usable antimicrobial spectrum that should be considered identical to that of cefoxitin. Disk diffusion and dilution test methods with cefmetazole correlated highly (r, greater than or equal to 0.95) with cefoxitin results. The recommended 30-micrograms cefmetazole disk interpretive breakpoints for susceptibility and resistance were greater than or equal to 18 mm (MIC, less than or equal to 8.0 micrograms/ml) and less than or equal to 14 mm (MIC, greater than or equal to 32 micrograms/ml), respectively. Cefmetazole and cefoxitin should be considered to be in the same antimicrobial spectrum class, requiring separate testing for other cephalosporins such as cephalothin, cefamandole, cefuroxime, and cefotetan. Recommended interpretive criteria performed well for fastidious organisms (Haemophilus influenzae, Neisseria meningitidis, and Branhamella catarrhalis) and for broth microdilution tests with anaerobes. Cefmetazole and cefoxitin broth disk elution tests for anaerobic bacteria produced higher rates of false susceptibility results. PMID:3097064

  17. Multivariate models for prediction of rheological characteristics of filamentous fermentation broth from the size distribution

    DEFF Research Database (Denmark)

    Petersen, Nanna; Stocks, S.; Gernaey, Krist

    2008-01-01

    fermentations conducted in 550 L pilot scale tanks were characterized with respect to particle size distribution, biomass concentration, and rheological properties. The rheological properties were described using the Herschel-Bulkley model. Estimation of all three parameters in the Herschel-Bulkley model (yield...... filamentous fermentations. It was therefore chosen to fix this parameter to the average value thereby decreasing the standard deviation of the estimates of the remaining theological parameters significantly. Using a PLSR model, a reasonable prediction of apparent viscosity (mu(app)), yield stress (tau......(y)), and consistency index (K), could be made from the size distributions, biomass concentration, and process information. This provides a predictive method with a high predictive power for the rheology of fermentation broth, and with the advantages over previous models that tau(y) and K can be predicted...

  18. The distribution and synonyms of Breutelia microdonta (Mitt.) Broth. : with additional notes on certain taxa of Breutelia

    OpenAIRE

    Griffin, Dana

    1992-01-01

    Breutelia microdonta (Mitt.) Broth., described originally from Brazil, is the oldest name for a disjunct widespread tropical species that includes as synonyms B. angustifolia Rehm. ex Sim of Southern Africa and B. merrillii Broth. of the Philippines. Breutelia kilaueae (C. Muell.) Broth. of Hawaii is considered a synonym of the austral Pacific species B. affinis (Hook.) Mitt., and B. brachyphylla Broth. of Ecuador is reduced to synonymy under the North Andean B. squarrosa Jaeg. Breutelia anac...

  19. Comparison of Carrot Broth- and Selective Todd-Hewitt Broth-Enhanced PCR Protocols for Real-Time Detection of Streptococcus agalactiae in Prenatal Vaginal/Anorectal Specimens ▿

    OpenAIRE

    Block, Timothy; Munson, Erik; Culver, Anne; Vaughan, Katharine; Hryciuk, Jeanne E.

    2008-01-01

    The reporting of accurate Streptococcus agalactiae screening results in a short time frame is of tremendous clinical benefit. A total of 203 consecutive primary vaginal/anorectal specimens were cultured in selective Todd-Hewitt broth (LIM broth) and with the StrepB carrot broth kit (carrot broth). One-day broth cultures were subjected to both centrifugation and clarification of a 500-μl aliquot prior to sample lysis (protocol A) and direct lysis of a 50-μl aliquot (protocol B). The lysates we...

  20. Temporal Characterization of Carrot Broth-Enhanced Real-Time PCR as an Alternative Means for Rapid Detection of Streptococcus agalactiae from Prenatal Anorectal and Vaginal Screenings ▿

    OpenAIRE

    Munson, Erik; Napierala, Maureen; Munson, Kimber L.; Culver, Anne; Hryciuk, Jeanne E.

    2010-01-01

    Analysis of overnight carrot broth culture using the BD GeneOhm StrepB assay (carrot broth-enhanced PCR) yields increased sensitivity compared to that of carrot broth culture alone for the detection of Streptococcus agalactiae. We investigated the prospect of reducing the carrot broth incubation time prior to PCR performance. In vitro experimentation demonstrated that carrot broth-enhanced PCR nominally detected 10 CFU S. agalactiae after 4 h of carrot broth incubation with competitive flora....

  1. Membrane-based recovery and dehydration of alcohols from fermentation broths - of materials and modules

    Science.gov (United States)

    Distillation combined with molecular sieve dehydration is the current state of the art for fuel grade ethanol production from fermentation broths. As the liquid biofuels industry transitions to lignocellulosic feedstocks, expands the end product portfolio to include other alcoho...

  2. Energy efficient recovery and dehydration of ethanol from fermentation broths by Membrane Assisted Vapor Stripping technology

    Science.gov (United States)

    Distillation combined with molecular sieve dehydration is the current state of the art for fuel grade ethanol production from fermentation broths. To improve the sustainability of bioethanol production, energy efficient separation alternatives are needed, particularly for lower ...

  3. In vitro Activity of Celery Essential Oil against Malassezia furfur

    OpenAIRE

    Chee, Hee Youn; Lee, Min Hee

    2009-01-01

    Antifungal activity of celery essential oil against Malassezia furfur was investigated using broth microdilution and vapor contact methods. Potent antifungal activity was evident using both methods. Fungicidal activity was revealed in the vapor contact method.

  4. Modeling of mixing for stirred bioreactors: 3. Mixing time for aerated simulated broths

    OpenAIRE

    Cascaval Dan; Oniscu Corneliu; Galaction Anca-Irina; Ungureanu Fiorina

    2002-01-01

    This paper presents the experiments on mixing efficiency for aerated media for a laboratory stirred bioreactor with a double turbine impeller. The effects of stirrer rotation speed, air volumetric flow rate and stirrer position on the shaft on mixing time for aerated water and simulated broths (CMCNa solutions) were analyzed. Compared to non-aerated broths, the results indicated that the variation of mixing time with the considered parameters is very different, due to the complex flow mechani...

  5. A new cyclopeptide with antifungal activity from the co-culture broth of two marine mangrove fungi.

    Science.gov (United States)

    Li, Chunyuan; Wang, Jinhua; Luo, Cuiping; Ding, Weijia; Cox, Daniel G

    2014-01-01

    A new cyclic tetrapeptide, cyclo-(L-leucyl-trans-4-hydroxy-L-prolyl-D-leucyl-trans-4-hydroxy-L-proline) (1), was isolated from the co-culture broth of two mangrove fungi Phomopsis sp. K38 and Alternaria sp. E33. The structure of 1 was determined by analysis of spectroscopic data and Marfey's analytic method. Primary bioassay demonstrated that compound 1 exhibited moderate to high inhibitory activity against four crop-threatening fungi including Gaeumannomyces graminis, Rhizoctonia cerealis, Helminthosporium sativum and Fusarium graminearum as compared with triadimefon. PMID:24571709

  6. Salting-out Extraction of 2,3-Butanediol from Jerusalem artichoke-based Fermentation Broth

    Institute of Scientific and Technical Information of China (English)

    DAI Jianying; ZHANG Yuanli; XIU Zhilong

    2011-01-01

    The removal of solid impurities and separation of target products from a fermentation broth is becoming more tedious with the utilization of lignocelluloses as source of substrate.2,3-Butanediol,an important chemical used widely is also a main product of sugar-based fermentation carried out by Klebsiella pneumoniae.In this study,we investigated the use of salting-out extraction(SOE) that employed a K2HPO4/ethanol system consisting of 21% ethanol and 17% K2HPO4(mass fraction) to separate 2,3-butanediol from the viscous Jerusalem artichoke-based fermentation broth.After SOE,about 98% of solid matters was removed,and the viscosity decreased from 72.5 mPa s in the original fermentation broth to 4.4 mPa s in the top phase.The partition coefficient and yield of 2,3-butanediol reached 13.4 and 99%,respectively,and 89% of soluble proteins was removed from the broth.The results showed that SOE is an efficient way for isolating 2,3-BD from a highly viscous fermentation broth by removing much of the solid matters within the broth.

  7. Two new cyclopeptides from the co-culture broth of two marine mangrove fungi and their antifungal activity

    Directory of Open Access Journals (Sweden)

    Song Huang

    2014-01-01

    Full Text Available Background: The strategy that co-cultivation two microorganisms in a single confined environment were recently developed to generate new active natural products. In the study, two new cyclic tetrapeptides, cyclo (D-Pro-L-Tyr-L-Pro-L-Tyr (1 and cyclo (Gly-L-Phe-L-Pro-L-Tyr (2 were isolated from the co-culture broth of two mangrove fungi Phomopsis sp. K38 and Alternaria sp. E33. Their antifungal activity against Candida albicans, Gaeumannomyces graminis, Rhzioctonia cerealis, Helminthosporium sativum and Fusarium graminearum was evaluated. Materials and Methods: Different column chromatographic techniques with different solvent systems were used to separate the constituents of the n-butyl alcohol extract of the culture broth. The structures of compounds 1 and 2 were identified by analysis of spectroscopic data (one-dimensional, two-dimensional - nuclear magnetic resonance, mass spectrometry and Marfey′s analytic method. Dilution method was used for the evaluation of antifungal activity. Results: Compounds 1 and 2 were identified as cyclo (D-Pro-L-Tyr-L-Pro-L-Tyr and cyclo (Gly-L-Phe-L-Pro-L-Tyr, respectively. Compounds 1 and 2 showed moderate to high antifungal activities as compared with the positive control. Conclusions: Compounds 1 and 2 are new cyclopeptides with moderate antifungal activity being worthy of consideration for the development and research of antifungal agents.

  8. Development of a selective broth medium for the detection of injured Campylobacter jejuni by capacitance monitoring.

    Science.gov (United States)

    Line, J Eric; Pearson, Kirsten G

    2003-10-01

    The purpose of these studies was to develop a conductimetric method for the rapid detection of Campylobacter jejuni. Numerous basal medium components were analyzed to develop a growth-enhancing broth medium for detection of freeze-injured Campylobacter cells using a conductimetric system. The final medium was composed of a modified Campy-Line agar from which the agar and triphenyltetrazolium chloride were removed and the amino acid, L-arginine was added. Pure isolates of C. jejuni. (frozen and thawed to produce stressed cells) were utilized to test the detection methodology. Monitoring of significant changes in the capacitance signal was found suitable for detection of Campylobacter proliferation. Using stressed pure cultures, Campylobacter growth was repeatedly detected at very low inoculum levels (about one cell per well). There was a direct linear relationship between detection times (DTs) and the initial inoculum level. For example, using a single strain, the mean DT (n = 20) at the 10 CFU/ml inoculum level was 28.6 h, with 100% of the inoculated wells detecting. The mean DTs at the 100, 1,000, and 10,000 CFU/ml inoculum levels were 24.9, 21.4, and 17.0 h, respectively. This study demonstrates that conductimetric methods can be utilized for the rapid detection of C. jejuni. PMID:14572208

  9. Detonation nanodiamonds for rapid detection of clinical isolates of Mycobacterium tuberculosis complex in broth culture media.

    Science.gov (United States)

    Soo, Po-Chi; Kung, Ching-Jen; Horng, Yu-Tze; Chang, Kai-Chih; Lee, Jen-Jyh; Peng, Wen-Ping

    2012-09-18

    Routinely used molecular diagnostic methods for mycobacterium identification are expensive and time-consuming. To tackle this problem, we develop a method to streamline identification of Mycobacterium tuberculosis complex (MTBC) in broth culture media by using detonation nanodiamonds (DNDs) as a platform to effectively capture the antigen secreted by MTBC which is cultured in BACTEC MGIT 960, followed by the analysis of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS). The 5 nm DNDs can capture the MTBC secretory antigen without albumin interference. With on diamond digestion, we confirm the DND captured antigen is cell filtrate protein 10 (CFP-10) because its Mascot analysis shows a score of 68. The dot blotting method further verifies a positive reaction with anti-CFP-10, indicating that CFP-10 is secreted in the medium of mycobacterium growth indicator tube (MGIT) and captured by DNDs. The minimal CFP-10 protein detection limit was 0.09 μg/mL. Furthermore, our approach can avoid the false-positive identification of MTBC by immunological methods due to cross-reactivity. Five hundred consecutive clinical specimens subjected to routine mycobacteria identification in hospital were used in this study, and the sensitivity of our method is 100% and the specificity is 98%. The analysis of each MTBC sample from culture solution can be finished within 1 h and thus shortens the turnaround time of MTBC identification of gold standard culture methods. In sum, DND MALDI-TOF MS for the detection of MTBC is rapid, specific, safe, reliable, and inexpensive. PMID:22905748

  10. The inhibitory effect of natural microflora of food on growth of Listeria monocytogenes in enrichment broths.

    Science.gov (United States)

    Al-Zeyara, Shaikha A; Jarvis, Basil; Mackey, Bernard M

    2011-01-31

    The aims of this study were to (i) compare the inhibitory effects of the natural microflora of different foods on the growth of Listeria monocytogenes during enrichment in selective and non-selective broths; (ii) to isolate and identify components of the microflora of the most inhibitory food; and (iii) to determine which of these components was most inhibitory to growth of L. monocytogenes in co-culture studies. Growth of an antibiotic-resistant marker strain of L. monocytogenes was examined during enrichment of a range of different foods in Tryptone Soya Broth (TSB), Half Fraser Broth (HFB) and Oxoid Novel Enrichment (ONE) Broth. Inhibition of L. monocytogenes was greatest in the presence of minced beef, salami and soft cheese and least with prepared fresh salad and chicken pâté. For any particular food the numbers of L. monocytogenes present after 24h enrichment in different broths increased in the order: TSB, HFB and ONE Broth. Numbers of L. monocytogenes recovered after enrichment in TSB were inversely related to the initial aerobic plate count (APC) in the food but with only a moderate coefficient of determination (R(2)) of 0.51 implying that microbial numbers and the composition of the microflora both influenced the degree of inhibition of L. monocytogenes. In HFB and ONE Broth the relationship between APC and final L. monocytogenes counts was weaker. The microflora of TSB after 24h enrichment of minced beef consisted of lactic acid bacteria, Brochothrix thermosphacta, Pseudomonas spp., Enterobacteriaceae, and enterococci. In co-culture studies of L. monocytogenes with different components of the microflora in TSB, the lactic acid bacteria were the most inhibitory followed by the Enterobacteriaceae. The least inhibitory organisms were Pseudomonas sp., enterococci and B. thermosphacta. In HFB and ONE Broth the growth of Gram-negative organisms was inhibited but lactic acid bacteria still reached high numbers after 24h. A more detailed study of the growth of

  11. Ergosterols from the Culture Broth of Marine Streptomyces anandii H41-59.

    Science.gov (United States)

    Zhang, Yang-Mei; Li, Hong-Yu; Hu, Chen; Sheng, Hui-Fan; Zhang, Ying; Lin, Bi-Run; Zhou, Guang-Xiong

    2016-01-01

    An actinomycete strain, H41-59, isolated from sea sediment in a mangrove district, was identified as Streptomyces anandii on the basis of 16S rDNA gene sequence analysis as well as the investigation of its morphological, physiological and biochemical characteristics. Three new ergosterols, ananstreps A-C (1-3), along with ten known ones (4-13), were isolated from the culture broth of this strain. The gross structures of these new compounds were elucidated on the basis of extensive analysis of spectroscopic data, including HR-ESI-MS, and NMR. The cytotoxicities of these isolates against human breast adenocarcinoma cell line MCF-7, human glioblastoma cell line SF-268, and human lung cancer cell line NCI-H460 and their antibacterial activities in inhibiting the growth of Candida albicans and some other pathogenic microorganisms were tested. Compounds 3-8, 10 and 11 displayed cytotoxicity with IC50 values in a range from 13.0 to 27.8 μg/mL. However, all the tested compounds showed no activity on C. albicans and other bacteria at the test concentration of 1 mg/mL with the paper disc diffusion method. PMID:27153073

  12. Ergosterols from the Culture Broth of Marine Streptomyces anandii H41-59

    Directory of Open Access Journals (Sweden)

    Yang-Mei Zhang

    2016-05-01

    Full Text Available An actinomycete strain, H41-59, isolated from sea sediment in a mangrove district, was identified as Streptomyces anandii on the basis of 16S rDNA gene sequence analysis as well as the investigation of its morphological, physiological and biochemical characteristics. Three new ergosterols, ananstreps A–C (1–3, along with ten known ones (4–13, were isolated from the culture broth of this strain. The gross structures of these new compounds were elucidated on the basis of extensive analysis of spectroscopic data, including HR-ESI-MS, and NMR. The cytotoxicities of these isolates against human breast adenocarcinoma cell line MCF-7, human glioblastoma cell line SF-268, and human lung cancer cell line NCI-H460 and their antibacterial activities in inhibiting the growth of Candida albicans and some other pathogenic microorganisms were tested. Compounds 3–8, 10 and 11 displayed cytotoxicity with IC50 values in a range from 13.0 to 27.8 μg/mL. However, all the tested compounds showed no activity on C. albicans and other bacteria at the test concentration of 1 mg/mL with the paper disc diffusion method.

  13. Biological evaluation of Phellinus linteus-fermented broths as anti-inflammatory agents.

    Science.gov (United States)

    Lin, Chun-Jung; Lien, Hsiu-Man; Chang, Hsiao-Yun; Huang, Chao-Lu; Liu, Jau-Jin; Chang, Yun-Chieh; Chen, Chia-Chang; Lai, Chih-Ho

    2014-07-01

    Phellinus linteus and its constituent hispolon induce potent anti-inflammatory activity in macrophages. Efficient production of the effective constituent and the biological function of P. linteus in the regulation of innate sensing have rarely been investigated. The aim of this study was to efficiently manufacture P. linteus-fermented broth containing the effective constituent, hispolon, and evaluate its immunoregulatory functions in macrophages. Four distinct fermented broths (PL1-4) and the medium dialyzate (MD) were prepared to screen suitable culture conditions for the mycelial growth of P. linteus. The P. linteus-fermented broth exhibited a dose-responsive inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production by murine macrophages. In addition, the P. linteus-fermented broths suppressed macrophage LPS-mediated nuclear factor (NF)-κB activity and tumor necrosis factor (TNF)-α. Among the tested samples from P. linteus, PL4 contained vast amounts of hispolon and showed the greatest anti-inflammatory activity in both the RAW264.7 cells and murine primary peritoneal exudate macrophages (PEMs). This study demonstrates that the purification of the effective constituent from P. linteus-fermented broth may enable the production of a potent therapeutic agent for anti-inflammation in macrophages. PMID:24503424

  14. A new anti-hypoxia functional agent Schizophyllum commune fermented broth and its biochemical mechanisms

    Institute of Scientific and Technical Information of China (English)

    Hao Limin; Lu Jike; Wu Tianyi; Ba Jianming; Guo Changjiang; Qian Ping; Yu Jianyong

    2013-01-01

    In order to overcome hypoxia induced sickness,a stable source of anti-hypoxia functional agents,Schizophyllum commune fermented broth has been developed in this study.Animal experiments were conducted to examine its anti-hypoxia activities and possible mechanisms involved.The acute hypoxic experiment showed that Schizophyllum commune fermented broth could significantly prolong the survival time of mice.The underlying mechanisms were associated with improved energy metabolism based on a study carried out in rats exposed to a low pressure chamber simulating the low pressure environment of 8000 m altitude.It was concluded that the Schizophyllum commune fermented broth was an effective anti-hypoxia functional agent and could be greatly beneficial to those living and working at high altitudes,such as people who reside in the reconstruction regions of Yushu.

  15. Nutrient Requirements of Renibacterium salmoninarum on Agar and in Broth Media.

    Science.gov (United States)

    Daly, J G; Stevenson, R M

    1993-07-01

    In well-aerated broth cultures, good growth of Renibacterium salmoninarum was obtained in a serum-free medium consisting of 1% peptone, 1% yeast extract, and 0.1% l-cysteine (PYC broth). In contrast, serum or charcoal is required for growth on agar medium. Charcoal treatment of broth media, either before bacterial inoculation or during growth, increased the growth of R. salmoninarum, whereas the surfactants Tween 20 and Tween 80 inhibited growth. l-Cysteine was essential for optimal growth. Other organic sulfur compounds, such as d-cysteine, l-methionine, homocysteine, homocysteine thiolactone, and reduced glutathione, supported only lower levels of growth, while cystine and dithiothreitol did not allow growth. PMID:16348993

  16. Invasive mycosis due to species of Blastobotrys in immunocompromised patients with reduced susceptibility to antifungals

    NARCIS (Netherlands)

    Kumar, A.; Babu, R.; Bijulal, S.; Abraham, M.; Sasidharan, P.; Kathuria, S.; Sharma, C.; Meis, J.F.G.M.; Chowdhary, A.

    2014-01-01

    Cases of invasive mycosis due to Blastobotrys serpentis and B. proliferans identified by sequencing in a preterm patient and a rhabdomyosarcoma patient, respectively, are reported. Both species revealed elevated fluconazole and echinocandin MICs by the CLSI broth microdilution method. Additionally,

  17. Effect of Pretreating the Broth of Hyaluronic Acid by Ultrasonic Wave%超声波对发酵液中提取透明质酸的影响

    Institute of Scientific and Technical Information of China (English)

    吴华昌; 马钦元; 邓静; 石岩昌; 由耀辉

    2009-01-01

    Objective:Taking the improvement of the extraction rate.Method:The effect of purification of Hyaluronic acid from broth by ethanol precipitation method was investigated by pretreating the broth using ultrasonic wave.Result: For the untreated broth,the extraction rate of Hyaluronic acid from the treated broth was improved by 38.6% under the condition of ultrasonic wave frequency 40kHz ,ultrasonic time 10min,temperature 15℃ and ultrasonic power 100W.A conclusion could be got that the extraction rate could be improved from the broth pretreated by ultrasonic wave.%目的:提高发酵液中透明质酸的提取率.方法:应用超声波预处理发酵液的方法对醇沉法提取透明质酸的影响进行研究.结果:在超声波功率100W、处理温度15℃、处理时间10min时,透明质酸提取率相对于未经超声波处理的发酵液提高了38.6%.因此,超声波预处理透明质酸发酵液可以达到提高透明质酸提取率的目的.

  18. Production of High-Viscosity Whey Broths by a Lactose-Utilizing Xanthomonas campestris Strain

    OpenAIRE

    Schwartz, Robert D.; Bodie, Elizabeth A.

    1985-01-01

    Xanthomonas campestris BB-1L was isolated by enrichment and selection by serial passage in a lactose-minimal medium. When BB-1L was subsequently grown in medium containing only 4% whey and 0.05% yeast extract, the lactose was consumed and broth viscosities greater than 500 cps at a 12 s−1 shear rate were produced. Prolonged maintenance in whey resulted in the loss of the ability of BB-1L to produce viscous broths in whey, indicating a reversion to preferential growth on whey protein, like the...

  19. Synergy of β-Lactams with Vancomycin against Methicillin-Resistant Staphylococcus aureus: Correlation of Disk Diffusion and Checkerboard Methods.

    Science.gov (United States)

    Sy, Cheng Len; Huang, Tsi-Shu; Chen, Chii Shiang; Chen, Yao-Shen; Tsai, Hung-Chin; Wann, Shue-Renn; Wu, Kuan-Sheng; Chen, Jui-Kuang; Lee, Susan Shin-Jung; Liu, Yung-Ching

    2016-03-01

    Modified disk diffusion (MDD) and checkerboard tests were employed to assess the synergy of combinations of vancomycin and β-lactam antibiotics for 59 clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and Mu50 (ATCC 700699). Bacterial inocula equivalent to 0.5 and 2.0 McFarland standard were inoculated on agar plates containing 0, 0.5, 1, and 2 μg/ml of vancomycin. Oxacillin-, cefazolin-, and cefoxitin-impregnated disks were applied to the surface, and the zones of inhibition were measured at 24 h. The CLSI-recommended checkerboard method was used as a reference to detect synergy. The MICs for vancomycin were determined using the Etest method, broth microdilution, and the Vitek 2 automated system. Synergy was observed with the checkerboard method in 51% to 60% of the isolates when vancomycin was combined with any β-lactam. The fractional inhibitory concentration indices were significantly lower in MRSA isolates with higher vancomycin MIC combinations (P oxacillin, 45.1% and 52.4% for cefazolin, and 43.1% and 52.4% for cefoxitin when combined with 0.5 and 2 μg/ml of vancomycin, respectively. Based on our study, the simple MDD method is not recommended as a replacement for the checkerboard method to detect synergy. However, it may serve as an initial screening method for the detection of potential synergy when it is not feasible to perform other labor-intensive synergy tests. PMID:26677253

  20. Studies on Concentration Process of Bacillus thuringiensis Fermentation Broth%苏云金芽孢杆菌发酵液浓缩工艺研究

    Institute of Scientific and Technical Information of China (English)

    周荣华; 崔怡宁; 饶犇; 廖先清; 刘芳; 陈伟

    2015-01-01

    通过对2种不同亚种的苏云金芽孢杆菌(Bacillus thuringiensis)发酵液进行浓缩工艺的研究,探索了不同温度对浓缩液有效成分的影响程度,比较了不同浓缩方式﹑不同干燥方式处理下有效成分的回收率. 结果表明,真空薄膜浓缩工艺的回收率要高于离心浓缩工艺23%~29%.%The concentration processes of two different subspecies of Bacillus thuringiensis fermentation broth were studied.The impact of different concentrate temperatures on the active ingredient of the fermentation broth was explored.The active ingredient recoveries of the fermentation broth deal with different concentration methods and different drying methods were compared.The results showed that the recovery by using vacuum thin film concentration was 23%~29% higher than concen-tration by centrifuging.

  1. Development of a multi-pathogen enrichment broth for simultaneous growth of five common foodborne pathogens.

    Science.gov (United States)

    Chen, Juan; Tang, Junni; Bhunia, Arun K; Tang, Cheng; Wang, Changting; Shi, Hui

    2015-01-01

    The objective of the present study was to formulate a multi-pathogen enrichment broth which could support the simultaneous growth of five common foodborne pathogens (Salmonella enterica, Staphylococcus aureus, Shigella flexneri, Listeria monocytogenes and Escherichia coli O157:H7). The formulated broth SSSLE was composed of potassium tellurite, bile salt, lithium chloride, and sodium chloride as growth-inhibitors; glucose, esculin, mannitol and sodium pyruvate as growth-promoters. Compared with the respective specific selective enrichment broths, the individual growth pattern of each target pathogen in SSSLE was equal, or even better, except in the case of S. flexneri. In mixed-culture experiments, the gram-negative bacteria showed higher growth capabilities than the gram-positive bacteria after 8-h enrichment; however, the cell numbers after 24-h enrichment indicated that SSSLE could support the concurrent growth of five target pathogens irrespective of whether pathogens were inoculated initially at equal or unequal levels. For natural food samples under the high background flora, the final cell numbers enriched in SSSLE for five targets were enough to be detected by multiplex PCR. In conclusion, SSSLE was capable of supporting the growth of five target pathogens concurrently. The new broth formulated in this study has the potential of saving time, efforts and costs in multi-pathogen enrichment procedures. PMID:26782652

  2. Isolation and Characterization of Exopolysaccharide with Immunomodulatory Activity from Fermentation Broth of Morchella Conica

    Directory of Open Access Journals (Sweden)

    Chao-an Su

    2013-01-01

    Full Text Available Background and the purpose of this study: Mushroom polysaccharides have traditionally been used for the prevention and treatment of a multitude of disorders like infectious illnesses, cancers and various autoimmune diseases. In vitro and in vivo studies suggest that certain polysaccharides affect immune system function. Morchella conica (M. conica is a species of rare edible mushroom whose multiple medicinal functions have been proven. Thus, the objective of this study is to isolate and characterize of exopolysaccharide from submerged mycelial culture of M. conica, and to evaluate its immunomodulatory activity.MethodsA water-soluble Morchella conica Polysaccharides (MCP were extracted and isolated from the fermentation broth of M. conica through a combination of DEAE-cellulose and Sephacryl S-300 HR chromatograph. NMR and IR spectroscopy has played a developing role in identification of polysaccharide with different structure and composition from fungal and plant sources, as well as complex glycosaminoglycans of animal origin. Thus, NMR and IR spectroscopy were used to analyze the chemical structure and composition of the isolated polysaccharide. Moreover, the polysaccharide was tested for its immunomodulatory activity at different concentrations using in vitro model.ResultsThe results showed that MCP may significantly modulate nitric oxide production in macrophages, and promote splenocytes proliferation. Analysis from HPLC, infrared spectra and nuclear magnetic resonance spectroscopy showed that MCP was a homogeneous mannan with an average molecular weight of approximately 81.2 kDa. The glycosidic bond links is [rightwards arrow]6-alpha-D-Man p-(1[rightwards arrow].ConclusionThe results suggested that the extracted MCP may modulate nitric oxide production in macrophages and promote splenocytes proliferation, and it may act as a potent immunomodulatory agent.

  3. Investigation of the chemical composition-antibacterial activity relationship of essential oils by chemometric methods.

    Science.gov (United States)

    Miladinović, Dragoljub L; Ilić, Budimir S; Mihajilov-Krstev, Tatjana M; Nikolić, Nikola D; Miladinović, Ljiljana C; Cvetković, Olga G

    2012-05-01

    The antibacterial effects of Thymus vulgaris (Lamiaceae), Lavandula angustifolia (Lamiaceae), and Calamintha nepeta (Lamiaceae) Savi subsp. nepeta var. subisodonda (Borb.) Hayek essential oils on five different bacteria were estimated. Laboratory control strain and clinical isolates from different pathogenic media were researched by broth microdilution method, with an emphasis on a chemical composition-antibacterial activity relationship. The main constituents of thyme oil were thymol (59.95%) and p-cymene (18.34%). Linalool acetate (38.23%) and β-linalool (35.01%) were main compounds in lavender oil. C. nepeta essential oil was characterized by a high percentage of piperitone oxide (59.07%) and limonene (9.05%). Essential oils have been found to have antimicrobial activity against all tested microorganisms. Classification and comparison of essential oils on the basis of their chemical composition and antibacterial activity were made by utilization of appropriate chemometric methods. The chemical principal component analysis (PCA) and hierachical cluster analysis (HCA) separated essential oils into two groups and two sub-groups. Thyme essential oil forms separate chemical HCA group and exhibits highest antibacterial activity, similar to tetracycline. Essential oils of lavender and C. nepeta in the same chemical HCA group were classified in different groups, within antibacterial PCA and HCA analyses. Lavender oil exhibits higher antibacterial ability in comparison with C. nepeta essential oil, probably based on the concept of synergistic activity of essential oil components. PMID:22389175

  4. Effective control of Listeria monocytogenes by combination of nisin formulated and slowly released into a broth system.

    Science.gov (United States)

    Chi-Zhang, Yundong; Yam, Kit L; Chikindas, Michael L

    2004-01-01

    In order to identify conditions for efficient food preservation by nisin, the sensitivity of Listeria monocytogenes to this preservative was studied under the following three model conditions: (1) the instantaneous addition of nisin into broth medium to simulate the formation of nisin in foods, (2) the slow delivery of nisin solution into broth medium using a pump to simulate the slow release of nisin from packaging materials to foods, (3) a combination of the two delivery methods. Based on the following results, we conclude that the antimicrobial effectiveness of nisin strongly depends on its mode of delivery. The instantaneous and slow methods for adding nisin inhibited L. monocytogenes, but over time of exposure, L. monocytogenes developed tolerance to nisin. Our data indicate that cells treated with instantaneously added nisin developed resistance to higher concentrations of nisin (200 IU/ml), compared to cells treated with slowly added nisin at the same total amount of the antimicrobial. Further studies indicated that nisin-tolerant cells recovered from treatments in which 200 IU/ml nisin was added instantaneously were likely to be mutants, which became resistant to the bacteriocin. In contrast, when 200 IU/ml of the antimicrobial was added slowly to the cells, only a temporary tolerance was developed; these cells became nisin-sensitive after passage through nisin-free medium. Due to the development of nisin-resistant cells, excessive amounts of nisin in the model system did not further inhibit L. monocytogenes. These results signify that excess nisin in foods does not necessarily improve the efficiency of controlling L. monocytogenes. Our data suggest that the combination of packaging material containing nisin used in conjunction with nisin-containing foods will provide the most effective means of preventing L. monocytogenes growth. PMID:14672827

  5. A simple one pot purification of bacterial amylase from fermented broth based on affinity toward starch-functionalized magnetic nanoparticle.

    Science.gov (United States)

    Paul, Tanima; Chatterjee, Saptarshi; Bandyopadhyay, Arghya; Chattopadhyay, Dwiptirtha; Basu, Semanti; Sarkar, Keka

    2015-08-18

    Surface-functionalized adsorbant particles in combination with magnetic separation techniques have received considerable attention in recent years. Selective manipulation on such magnetic nanoparticles permits separation with high affinity in the presence of other suspended solids. Amylase is used extensively in food and allied industries. Purification of amylase from bacterial sources is a matter of concern because most of the industrial need for amylase is met by microbial sources. Here we report a simple, cost-effective, one-pot purification technique for bacterial amylase directly from fermented broth of Bacillus megaterium utilizing starch-coated superparamagnetic iron oxide nanoparticles (SPION). SPION was prepared by co-precipitation method and then functionalized by starch coating. The synthesized nanoparticles were characterized by transmission electron microscopy (TEM), a superconducting quantum interference device (SQUID, zeta potential, and ultraviolet-visible (UV-vis) and Fourier-transform infrared (FTIR) spectroscopy. The starch-coated nanoparticles efficiently purified amylase from bacterial fermented broth with 93.22% recovery and 12.57-fold purification. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the molecular mass of the purified amylase was 67 kD, and native gel showed the retention of amylase activity even after purification. Optimum pH and temperature of the purified amylase were 7 and 50°C, respectively, and it was stable over a range of 20°C to 50°C. Hence, an improved one-pot bacterial amylase purification method was developed using starch-coated SPION. PMID:24840788

  6. Influence of different susceptibility testing methods and media on determination of the relevant fluconazole minimum inhibitory concentrations for heavy trailing Candida isolates with low-high phenotype.

    Science.gov (United States)

    Alp, Sehnaz; Sancak, Banu; Hascelik, Gulsen; Arikan, Sevtap

    2010-11-01

    We investigated the incidence of trailing growth with fluconazole in 101 clinical Candida isolates (49 C. albicans and 52 C. tropicalis) and tried to establish the convenient susceptibility testing method and medium for fluconazole minimum inhibitory concentration (MIC) determination. MICs were determined by CLSI M27-A2 broth microdilution (BMD) and Etest methods on RPMI-1640 agar supplemented with 2% glucose (RPG) and on Mueller-Hinton agar supplemented with 2% glucose and 0.5 μg ml(-1) methylene blue (GMB). BMD and Etest MICs were read at 24 and 48 h, and susceptibility categories were compared. All isolates were determined as susceptible with BMD, Etest-RPG and Etest-GMB at 24 h. While all isolates were interpreted as susceptible at 48 h on Etest-RPG and Etest-GMB, one C. albicans isolate was interpreted as susceptible-dose dependent (S-DD) and two C. tropicalis isolates were interpreted as resistant with BMD. On Etest-RPG, trailing growth caused widespread microcolonies within the inhibition zone and resulted in confusion in MIC determination. On Etest-GMB, because of the nearly absence of microcolonies within the zone of inhibition, MICs were evaluated more easily. We conclude that, for the determination of fluconazole MICs of trailing Candida isolates, the Etest method has an advantage over BMD and can be used along with this reference method. Moreover, GMB appears more beneficial than RPG for the fluconazole Etest. PMID:19563491

  7. Evaluation of nanofiltration for the purification of an organic acid fermentation broth

    OpenAIRE

    Bouchoux, Antoine; Lutin, F.; Roux-De Balmann, Hélène

    2006-01-01

    This study focuses on a possible elimination of glucose from a sodium lactate fermentation broth by nanofiltration (NF). The results obtained with single-solute solutions indicate that glucose is clearly more retained that sodium lactate. Nevertheless, glucose retention strongly decreases when the two solutes are present in the same solution and the separation becomes unachievable. Such a incidence of a solute on another in NF has rarely been reported before.

  8. The influences of fish infusion broth on the biogenic amines formation by lactic acid bacteria

    OpenAIRE

    Esmeray Küley; Fatih Özogul; Esra Balikçi; Mustafa Durmus; Deniz Ayas

    2013-01-01

    The influences of fish infusion decarboxylase broth (IDB) on biogenic amines (BA) formation by lactic acid bacteria (LAB) were investigated. BA productions by single LAB strains were tested in five different fish (anchovy, mackerel, white shark, sardine and gilthead seabream) IDB. The result of the study showed that significant differences in ammonia (AMN) and BA production were observed among the LAB strains in fish IDB (p < 0.05). The highest AMN and TMA production by LAB strains were obser...

  9. UV-Heat Treatments for the Control of Foodborne Microbial Pathogens in Chicken Broth

    Directory of Open Access Journals (Sweden)

    M. Gouma

    2015-01-01

    Full Text Available This investigation established the process criteria for using UV-C light and mild heat (UV-H treatment to inactivate 5-Log10 cycles (performance criterion of common foodborne pathogen populations, Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, when inoculated in chicken broth. To define the target microorganism and the proper UV-H treatment conditions (including UV dose, treatment time, and temperature that would achieve the stated performance criterion, mathematical equations based on Geeraerd’s model were developed for each microorganism. For the sake of comparison, inactivation equations for heat treatments were also performed on the same chicken broth and for the same microorganisms. L. monocytogenes was the most UV-H resistant microorganism at all temperatures, requiring a UV dose between 6.10 J/mL (5.6 min and 2.26 J/mL (2.09 min to achieve 5-Log10 reductions. In comparison with UV treatments at room temperatures, the combination of UV and mild heat allowed both the UV dose and treatment time to be reduced by 30% and 63% at 55°C and 60°C, respectively. Compared to heat treatments, the UV-H process reduced the heating time for 5-Log10 reductions of all the investigated microorganisms in chicken broth from 20-fold to 2-fold when the operating temperature varied from 53 to 60°C.

  10. Techniques for anaerobic susceptibility testing.

    Science.gov (United States)

    Thornsberry, C

    1977-03-01

    Minimal inhibitory concentrations (MICs) of antimicrobial agents for anaerobic bacteria can be determined by agar dilution and broth dilution (including microdilution) techniques. If MICs are not determined routinely, the disk broth or category methods are recommended for routine use. The Bauer-Kirby disk diffusion method and its interpretative standards should not be used for anaerobes. PMID:850089

  11. Pharmacodynamic Interaction of Quercus infectoria Galls Extract in Combination with Vancomycin against MRSA Using Microdilution Checkerboard and Time-Kill Assay.

    Science.gov (United States)

    Basri, Dayang Fredalina; Khairon, Radhiah

    2012-01-01

    The galls of Quercus infectoria Olivier possess astringent properties which helps in the tightening of the vaginal epithelium in the post-natal period. The present study aimed to observe the time-kill kinetics of the acetone and methanol extracts of gall of Q. infectoria in combination with vancomycin against two methicillin-resistant Staphylococcus aureus (MRSA) strains; ATCC 33591 and MU 9495 (laboratory-passaged strain). Minimum inhibitory concentration (MIC) of the extracts were determined using microdilution technique whereas the checkerboard and time-kill kinetics were employed to verify the synergistic effects of treatment with vancomycin. The FIC index value of the combinations against both MRSA strains showed that the interaction was synergistic (FIC index infectoria with vancomycin were synergistic according to FIC index values. The time-kill curves showed that the interaction was additive with a more rapid killing rate but, which did not differ significantly with vancomycin. PMID:22899953

  12. Evaluation of Three Commercial Broth Media for Pigment Detection and Identification of a Group B Streptococcus (Streptococcus agalactiae)▿

    OpenAIRE

    Carvalho, Maria da Glória; Facklam, Richard; Jackson, Delois; Beall, Bernard; McGee, Lesley

    2009-01-01

    Detection of group B Streptococcus (GBS) strains at various bacterial concentrations was evaluated using three pigment-producing broth media. At 103 CFU/ml, StrepB carrot broth (SBCB), Granada instant liquid biphasic (IGLB), and Northeast Laboratory GBS screening medium (NEL-GBS) showed 100% detection, but at the lower bacterial counts, SBCB and IGLB were more sensitive than NEL-GBS after 24 h.

  13. Controlled evaluation of supplemented peptone and Bactec blood culture broths for the detection of bacteremia and fungemia.

    OpenAIRE

    Reimer, L G; McDaniel, J D; Mirrett, S.; Reller, L B; Wang, W L

    1985-01-01

    Comparison of conventional blood culture media with newer formulations of Bactec media for radiometric detection are lacking. Therefore, we compared the yield and speed of detection of clinically important microorganisms with supplemented peptone broth (SPB) and Bactec aerobic (6B) and anaerobic (7C or 7D) broths in 7,627 blood samples from adult patients. Acridine orange stains from SPB, radiometric readings from Bactec, and routine subcultures from all bottles were done at the same time int...

  14. Gatifloxacin (AM-1155, CG 5501) susceptibility testing interpretive criteria and quality control guidelines for dilution and disk (5-microgram) diffusion methods. The Quality Control Study Group.

    Science.gov (United States)

    Jones, R N; Kugler, K C; Erwin, M E; Biedenbach, D J; Beach, M L; Pfaller, M A

    1999-04-01

    Gatifloxacin (formerly AM-1155 and CG5501), a new 8-methoxy fluoroquinolone, has an expanded spectrum of activity against Gram-positive cocci and some anaerobic bacteria. This compound was tested against 600 recent clinical strains of rapidly growing aerobic species to establish susceptibility testing interpretive criteria for the reference broth microdilution and standardized disk (5-microgram) diffusion methods of the National Committee for Clinical Laboratory Standards (NCCLS). These strains included 285 Enterobacteriaceae (17 species), 165 staphylococci, 49 enterococci, and 101 nonfermentative Gram-negative bacilli. Based on achievable serum levels with projected gatifloxacin dosing regimens, MIC break points of or = 18 mm) for susceptibility and > or = 8 micrograms/mL (< or = 14 mm) for resistance were selected. The absolute agreement between tests was 94.3% with no very major false-resistant errors. The quality control ranges (MIC and zone diameters) for the NCCLS recommended strains were determined in a nine-laboratory NCCLS protocol as follows: Escherichia coli ATCC 25922 = 0.008-0.03 microgram/mL and 31-37 mm; Enterococcus faecalis ATCC 29212 = 0.12-1 microgram/mL; Pseudomonas aeruginosa ATCC 27853 = 0.5-2 micrograms/ml and 21-27 mm; Staphylococcus aureus ATCC 25923 = 27-33 mm and S. aureus ATCC 29213 = 0.03-0.12 microgram/mL. Gatifloxacin appears to be a promising new fluoroquinolone with acceptable susceptibility testing methods for routine clinical laboratory use. PMID:10212751

  15. Automated harvesting and 2-step purification of unclarified mammalian cell-culture broths containing antibodies.

    Science.gov (United States)

    Holenstein, Fabian; Eriksson, Christer; Erlandsson, Ioana; Norrman, Nils; Simon, Jill; Danielsson, Åke; Milicov, Adriana; Schindler, Patrick; Schlaeppi, Jean-Marc

    2015-10-30

    Therapeutic monoclonal antibodies represent one of the fastest growing segments in the pharmaceutical market. The growth of the segment has necessitated development of new efficient and cost saving platforms for the preparation and analysis of early candidates for faster and better antibody selection and characterization. We report on a new integrated platform for automated harvesting of whole unclarified cell-culture broths, followed by in-line tandem affinity-capture, pH neutralization and size-exclusion chromatography of recombinant antibodies expressed transiently in mammalian human embryonic kidney 293T-cells at the 1-L scale. The system consists of two bench-top chromatography instruments connected to a central unit with eight disposable filtration devices used for loading and filtering the cell cultures. The staggered parallel multi-step configuration of the system allows unattended processing of eight samples in less than 24h. The system was validated with a random panel of 45 whole-cell culture broths containing recombinant antibodies in the early profiling phase. The results showed that the overall performances of the preparative automated system were higher compared to the conventional downstream process including manual harvesting and purification. The mean recovery of purified material from the culture-broth was 66.7%, representing a 20% increase compared to that of the manual process. Moreover, the automated process reduced by 3-fold the amount of residual aggregates in the purified antibody fractions, indicating that the automated system allows the cost-efficient and timely preparation of antibodies in the 20-200mg range, and covers the requirements for early in vitro and in vivo profiling and formulation of these drug candidates. PMID:26431859

  16. Green synthesis of palladium nanoparticles using broth of Cinnamomum camphora leaf

    Energy Technology Data Exchange (ETDEWEB)

    Yang Xin; Li Qingbiao, E-mail: kelqb@xmu.edu.cn; Wang Huixuan; Huang Jiale; Lin Liqin; Wang Wenta; Sun Daohua; Su Yuanbo; Opiyo, James Berya; Hong Luwei; Wang Yuanpeng; He Ning; Jia Lishan [College of Chemistry and Chemical Engineering, Xiamen University, Department of Chemical and Biochemical Engineering (China)

    2010-06-15

    The development of dependable, environmentally benign processes for the synthesis of nanoscale materials is an important aspect of nanotechnology. In the present study, we report one-pot biogenic fabrication of palladium nanoparticles by a simple procedure using broth of Cinnamomum camphora leaf without extra surfactant, capping agent, and/or template. The mean size of palladium nanoparticles, ranging from 3.2 to 6.0 nm, could be facilely controlled by merely varying the initial concentration of the palladium ions. The polyols components and the heterocyclic components were believed to be responsible for the reduction of palladium ions and the stabilization of palladium nanoparticles, respectively.

  17. Ultra-high-performance liquid chromatography fingerprinting method for chemical screening of metabolites in cultivation broth

    Czech Academy of Sciences Publication Activity Database

    Kameník, Zdeněk; Hadacek, F.; Marečková, M.; Ulanová, Dana; Kopecký, J.; Chobot, V.; Plháčková, Kamila; Olšovská, Jana

    2010-01-01

    Roč. 1217, č. 51 (2010), s. 8016-8025. ISSN 0021-9673 R&D Projects: GA MŠk 2B08064 Institutional research plan: CEZ:AV0Z50200510 Keywords : UHPLC * high-throughput fingerprinting * secondary metabolites Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 4.194, year: 2010

  18. Detection of Burkholderia pseudomallei in Sputum using Selective Enrichment Broth and Ashdown's Medium at Kampong Cham Provincial Hospital, Cambodia.

    Science.gov (United States)

    Nhem, Somary; Letchford, Joanne; Meas, Chea; Thann, Sovanndeth; McLaughlin, James C; Baron, Ellen Jo; West, T Eoin

    2014-01-01

    Melioidosis, infection caused by Burkholderia pseudomallei, is increasingly reported in Cambodia. We hypothesized that implementation of an enhanced sputum testing protocol in a provincial hospital diagnostic microbiology laboratory would increase detection of B. pseudomallei. We tested 241 sputum specimens that were deemed acceptable for culture, comparing culture in selective enrichment broth followed by sub-culture on Ashdown's medium to standard culture methods. Two specimens (0.8%) were positive for B. pseudomallei using the enhanced protocol whereas one specimen (0.4%) was positive using standard methods. Given the low numbers of positive specimens, we could not conclusively determine the utility of the enhanced sputum testing protocol. However, the ramifications of identification of  B. pseudomallei are substantial, and the benefit of the enhanced testing protocol may be more apparent in patients selected based on risk factors and clinical presentation. Promoting clinician awareness of the infection and encouraging utilization of diagnostic microbiology services are also likely to be important factors in facilitating identification of melioidosis. PMID:25717370

  19. Optimization of DsbA Purification from Recombinant Escherichia coil Broth Using Box-Behnken Design Methodology

    Institute of Scientific and Technical Information of China (English)

    LUO Man; GUAN Yixin; YAO Shanjing

    2013-01-01

    Disulfide bond formation protein A(DsbA)is one of the important helper proteins for folding in protein synthesis in vivo.In this study,purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method,a statistic-based design of experiments.The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem,which takes the protein recovery,purification efficiency and throughput of ion-exchange chromatography into account.After the optimization,protein recovery of 96.8% and purity higher than 95% DsbA was achieved,and the productivity was (377.9±1.7)mg soluble DsbA per liter broth.The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856,a mutant of DsbA.The DsbA was preliminarily applied to the refolding of denatured lysozyme m vitro.

  20. Crude oil biodegradation aided by biosurfactants from Pseudozyma sp. NII 08165 or its culture broth.

    Science.gov (United States)

    Sajna, Kuttuvan Valappil; Sukumaran, Rajeev Kumar; Gottumukkala, Lalitha Devi; Pandey, Ashok

    2015-09-01

    The aim of this work was to evaluate the biosurfactants produced by the yeast Pseudozyma sp. NII 08165 for enhancing the degradation of crude oil by a model hydrocarbon degrading strain, Pseudomonas putida MTCC 1194. Pseudozyma biosurfactants were supplemented at various concentrations to the P. putida culture medium containing crude oil as sole carbon source. Supplementation of the biosurfactants enhanced the degradation of crude oil by P. putida; the maximum degradation of hydrocarbons was observed with a 2.5 mg L(-1) supplementation of biosurfactants. Growth inhibition constant of the Pseudozyma biosurfactants was 11.07 mg L(-1). It was interesting to note that Pseudozyma sp. NII 08165 alone could also degrade diesel and kerosene. Culture broth of Pseudozyma containing biosurfactants resulted up to ∼46% improvement in degradation of C10-C24 alkanes by P. putida. The enhancement in degradation efficiency of the bacterium with the culture broth supplementation was even more pronounced than that with relatively purer biosurfactants. PMID:25985416

  1. High pressure inactivation of Pseudomonas in black truffle - comparison with Pseudomonas fluorescens in tryptone soya broth

    Science.gov (United States)

    Ballestra, Patricia; Verret, Catherine; Cruz, Christian; Largeteau, Alain; Demazeau, Gerard; El Moueffak, Abdelhamid

    2010-03-01

    Pseudomonas is one of the most common genera in black Perigord truffle. Its inactivation by high pressure (100-500 MPa/10 min) applied on truffles at sub-zero or low temperatures was studied and compared with those of Pseudomonas fluorescens in tryptone soya broth. Pressurization of truffles at 300 MPa/4 °C reduced the bacterial count of Pseudomonas by 5.3 log cycles. Higher pressures of 400 or 500 MPa, at 4 °C or 20 °C, allowed us to slightly increase the level of destruction to the value of ca. 6.5 log cycles but did not permit us to completely inactivate Pseudomonas. The results showed a residual charge of about 10 CFU/g. Pressure-shift freezing of truffles, which consists in applying a pressure of 200 MPa/-18 °C for 10 min and then quickly releasing this pressure to induce freezing, reduced the population of Pseudomonas by 3.3 log cycles. The level of inactivation was higher than those obtained with conventional freezing. Endogenous Pseudomonas in truffle was shown to be more resistant to high pressure treatments than P. fluorescens used for inoculation of broths.

  2. Determination of direct photolysis rate constants and OH radical reactivity of representative odour compounds in brewery broth using a continuous flow-stirred photoreactor

    Science.gov (United States)

    Jürgens, Marion; Jacob, Fritz; Ekici, Perihan; Friess, Albrecht; Parlar, Harun

    A method based on photolysis was developed for the appropriate treatment of organic pollutants in air exhausting from breweries upon wort decoction, and thereby causing smell nuisance. A continuous flow stirred photoreactor was built-up exclusively, allowing OH radicals to react with selected odorous compounds contained in exhaust vapours, such as: 2-methylpropanal, 3-methylbutanal, 2-methylbutanal, 3-methyl-1-butanol, n-hexanal, 2-methylbutyl isobutyrate, 2-undecanone, phenyl acetaldehyde, myrcene, limonene, linalool, humulene, dimethylsulphide, and dimethyltrisulphide. These substances were quantified in brewery broth before and after UV irradiation using high-resolution gas chromatography-mass spectrometry (HRGC-MS). For odour analysis, high-resolution gas chromatography-flame ionisation detection (HRGC-FID) coupled with sensory methods was used. Determined quantum yields of about 10 -3 for phenyl acetaldehyde, myrcene, and humulene pointed out that direct photolysis contributed to their decay. Quantum yields of below 10 -4 for the other substances indicated that UV irradiation did not contribute significantly to their degradation processes. Hydroxyl radical reaction rate constants and Henry constants of organic compounds were also measured. Substances accompanied with low Henry constants converted rapidly, whereas those with higher ones, relatively slowly. Determined aroma values concluded that after UV-H 2O 2 treatment, only dimethylsulphide and myrcene remained as important odorous compounds, but in significantly reduced concentrations. The UV-H 2O 2 treatment of brewery broth has been proved effective to reduce smell-irritating substances formed upon wort decoction.

  3. Evaluation of rapid alternative methods for drug susceptibility testing in clinical isolates of Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Luciano Mengatto

    2006-08-01

    Full Text Available A study was carried out to compare the performance of a commercial method (MGIT and four inexpensive drug susceptibility methods: nitrate reductase assay (NRA, microscopic observation drug susceptibility (MODS assay, MTT test, and broth microdilution method (BMM. A total of 64 clinical isolates of Mycobacterium tuberculosis were studied. The Lowenstein-Jensen proportion method (PM was used as gold standard. MGIT, NRA, MODS, and MTT results were available on an average of less than 10 days, whereas BMM results could be reported in about 20 days. Most of the evaluated tests showed excellent performance for isoniazid and rifampicin, with sensitivity and specificity values > 90%. With most of the assays, sensitivity for ethambutol was low (62-87% whereas for streptomycin, sensitivity values ranged from 84 to 100%; NRA-discrepancies were associated with cultures with a low proportion of EMB-resistant organisms while most discrepancies with quantitative tests (MMT and BMM were seen with isolates whose minimal inhibitory concentrations fell close the cutoff. MGIT is reliable but still expensive. NRA is the most inexpensive and easiest method to perform without changing the organization of the routine PM laboratory performance. While MODS, MTT, and BMM, have the disadvantage from the point of view of biosafety, they offer the possibility of detecting partial resistant strains. This study shows a very good level of agreement of the four low-cost methods compared to the PM for rapid detection of isoniazid, rifampicin and streptomycin resistance (Kappa values > 0.8; more standardization is needed for ethambutol.

  4. Comparison of Enrichment Broths for Supporting Growth of Shiga Toxin-Producing Escherichia coli.

    Science.gov (United States)

    Stromberg, Zachary R; Lewis, Gentry L; Marx, David B; Moxley, Rodney A

    2015-08-01

    Detection of Shiga toxin-producing Escherichia coli (STEC) in complex sample matrices remains challenging. In an attempt to improve detection, nonselective and selective enrichment broths were compared as follows: (1) trypticase soy broth (TSB) was compared with TSB plus novobiocin, vancomycin, rifampicin, bile salts, and potassium tellurite (TSB-NVRBT) for supporting growth of STEC in pure culture; (2) E. coli broth (EC), TSB, and TSB plus bile salts (mTSB) were compared for enrichment of STEC O26, O45, O103, O104, O111, O121, O145, and O157 (STEC-8) in inoculated cattle fecal samples; (3) EC, TSB, and mTSB were compared for the detection of STEC-8 in inoculated cattle fecal samples. Fecal samples were inoculated with wild-type STEC-8 or nalidixic acid- or rifampicin-resistant derivatives of the same strains at 100, 1000, or 10,000 colony-forming units per gram (CFU/g) of feces. In pure culture, the mean STEC CFU/mL following enrichment in TSB was 1.17 log10 greater than that in TSB-NVRBT (P < 0.05). In inoculated fecal samples, EC enrichment yielded growth of STEC-8 (6.42 log10 CFU/g) that was significantly greater than in TSB (6.23 log10 CFU/g; P < 0.05), and numerically but not significantly greater than in mTSB (6.37 log10 CFU/g; P = 0.60). Wild-type STEC strains were detected in 43.8 % (21/48) of the samples enriched in EC and mTSB compared to 27.1 % (13/48) of the samples enriched in TSB (P = 0.15). Overall, STEC grew significantly better when enriched in EC compared to TSB. Modification of TSB by the addition of bile salts improved the growth and detection of STEC compared to TSB alone. PMID:25917502

  5. Pharmacodynamic Interaction of Quercus infectoria Galls Extract in Combination with Vancomycin against MRSA Using Microdilution Checkerboard and Time-Kill Assay

    Directory of Open Access Journals (Sweden)

    Dayang Fredalina Basri

    2012-01-01

    Full Text Available The galls of Quercus infectoria Olivier possess astringent properties which helps in the tightening of the vaginal epithelium in the post-natal period. The present study aimed to observe the time-kill kinetics of the acetone and methanol extracts of gall of Q. infectoria in combination with vancomycin against two methicillin-resistant Staphylococcus aureus (MRSA strains; ATCC 33591 and MU 9495 (laboratory-passaged strain. Minimum inhibitory concentration (MIC of the extracts were determined using microdilution technique whereas the checkerboard and time-kill kinetics were employed to verify the synergistic effects of treatment with vancomycin. The FIC index value of the combinations against both MRSA strains showed that the interaction was synergistic (FIC index <0.5. Time-kill assays showed the bactericidal effect of the combination treatment at 1/8XMIC of the extract and 1/8XMIC of vancomycin, were respectively at 7.2±0.28 hr against ATCC 33591 compared to complete attenuation of the growth of the same strain after 8 hr of treatment with vancomycin alone. In conclusion, the combination extracts of Q. infectoria with vancomycin were synergistic according to FIC index values. The time-kill curves showed that the interaction was additive with a more rapid killing rate but, which did not differ significantly with vancomycin.

  6. Pre-treatment step with Leuconostoc mesenteroides or L. pseudomesenteroides strains removes furfural from Zymomonas mobilis ethanolic fermentation broth

    Science.gov (United States)

    Furfural (furan-2-carboxaldehyde), formed during dilute acid hydrolysis of biomass, is an inhibitor of growth and ethanol production by Zymomonas mobilis. The present study used a biological pre-treatment to reduce that amount of furfural in a model biofuel fermentation broth. The pre-treatment in...

  7. Purification of chondroitin precursor from Escherichia coli K4 fermentation broth using membrane processing.

    Science.gov (United States)

    Schiraldi, Chiara; Carcarino, Immacolata Loredana; Alfano, Alberto; Restaino, Odile Francesca; Panariello, Andrea; De Rosa, Mario

    2011-04-01

    Recently the possibility of producing the capsular polysaccharide K4, a fructosylated chondroitin, in fed-batch experiments was assessed. In the present study, a novel downstream process to obtain chondroitin from Escherichia coli K4 fermentation broth was developed. The process is simple, scalable and economical. In particular, downstream procedures were optimized with a particular aim of purifying a product suitable for further chemical modifications, in an attempt to develop a biotechnological platform for chondroitin sulfate production. During process development, membrane devices (ultrafiltration/diafiltration) were exploited, selecting the right cassette cut-offs for different phases of purification. The operational conditions (cross-flow rate and transmembrane pressure) used for the process were determined on an ÄKTA cross-flow instrument (GE Healthcare, USA), a lab-scale automatic tangential flow filtration system. In addition, parameters such as selectivity and throughput were calculated based on the analytical quantification of K4 and defructosylated K4, as well as the major contaminants. The complete downstream procedure yielded about 75% chondroitin with a purity higher than 90%. PMID:21381202

  8. Urokinase Separation from Cell Culture Broth of a Human Kidney Cell Line

    Directory of Open Access Journals (Sweden)

    Vibha Bansal, Pradip K. Roychoudhury, Ashok Kumar

    2007-01-01

    Full Text Available A single step ion-exchange chromatography on a sulfo-propyl (SP- Sepharose column was performed to separate both the high molecular weight (HMW- and low molecular weight (LMW- forms of enzymatically active urokinase type plasminogen activator from human kidney (HT1080 cell culture media. The level of urokinase secreted by the cell line reached to about 145 Plough units/ml culture broth within 48 h of cultivation. The conditioned cell culture media was applied directly to the column without any prior concentration steps. Polyacrylamide gel electrophoresis of the column eluates in the presence of sodium dodecyl sulphate showed that the cell line secretes three forms of two-chain high molecular weight (HMW urokinase of molecular weights (Mr 64,000, 60,900 and 55,000. In addition, two low molecular weight (LMW forms of Mr 22,000 and 20,000; proteolytic cleavage products of HMW, were also found. The HMW and LMW forms had intrinsic plasminogen dependent proteolytic activity as judged by zymographic analysis. The specific activity of the pooled peak fractions increased (approximately 93-fold to values as high as 1481 Plough units/ mg protein. Both HMW as well as LMW forms were obtained in significantly high yields.

  9. The influences of fish infusion broth on the biogenic amines formation by lactic acid bacteria

    Directory of Open Access Journals (Sweden)

    Esmeray Küley

    2013-01-01

    Full Text Available The influences of fish infusion decarboxylase broth (IDB on biogenic amines (BA formation by lactic acid bacteria (LAB were investigated. BA productions by single LAB strains were tested in five different fish (anchovy, mackerel, white shark, sardine and gilthead seabream IDB. The result of the study showed that significant differences in ammonia (AMN and BA production were observed among the LAB strains in fish IDB (p < 0.05. The highest AMN and TMA production by LAB strains were observed for white shark IDB. The all tested bacteria had decarboxylation activity in fish IDB. The uppermost accumulated amines by LAB strains were tyramine (TYM, dopamine, serotonin and spermidine. The maximum histamine production was observed in sardine (101.69 mg/L and mackerel (100.84 mg/L IDB by Leuconostoc mesenteroides subsp. cremoris and Pediococcus acidophilus, respectively. Lactobacillus delbrueckii subsp. lactis and Pediococcus acidophilus had a high TYM producing capability (2943 mg/L and 1157 mg/L in sardine IDB.

  10. Performance of serum-free broth media for growth of Renibacterium salmoninarum.

    Science.gov (United States)

    Starliper, C E; Schill, W B; Mathias, J

    1998-09-11

    Growth of Renibacterium salmoninarum was compared in 14 different broth media; 13 serum-free, and 1 that contained newborn calf serum, KDM2+M. Supplementation with 1% v/v R. salmoninarum MCO4M metabolite was evaluated for 6 of the media that do not utilize it as part of their ingredients. Viable cells were enumerated on Days 10, 20, and 30 post inoculation to evaluate performance. The experiment was repeated 3 times using high, low, and medium (trials 1 to 3, respectively) cell concentrations as inoculum. In general there was no optimal medium and all performed well. The choice of which to employ depends on the ease of preparation and presence of certain ingredients that might affect subsequent assays. In trials 2 and 3, the pH was estimated using test papers at the same time as cells were counted. Maximum pH increase occurred with KDM2+M and those media containing charcoal. For most media, a simple pH determination could be used as a means to check that growth has occurred in a culture, particularly if charcoal was added directly to the media and a visual inspection could not be made to detect growth. PMID:9789976

  11. 毛锦藓科中国新记录种——卷边麻锦藓%Taxithelium kerianum (Broth.) Broth.(Pylaisiadelphaceae, Bryophyta)Reported New to China

    Institute of Scientific and Technical Information of China (English)

    章博远; 魏倩倩; 韦玉梅; 贾渝; 朱瑞良; 王幼芳

    2015-01-01

    报道了产自广西的毛锦藓科1中国新记录种——卷边麻锦藓[Taxithelium kerianum (Broth.)Broth.],提供了该种详细的形态描述和图版,并编制了麻锦藓属的分种检索表,明确了卷边麻锦藓与中国分布的麻锦藓属其它种类之间的形态差别.该种主要分布在南亚及其附近地区,为典型的热带藓类.

  12. Evaluation of the disc diffusion method with a comparison study for fluconazole susceptibility of Candida strains

    Institute of Scientific and Technical Information of China (English)

    2003-01-01

    Objective To performance susceptibility testing of antifungal agents. Due to the increasing number of resistant strains, susceptibility testing of antifungal agents is gaining importance. Methods We compared the results of standard macrotube dilution reference method with two different microdilution methods, as well as the disc diffusion method in order to test the susceptibility of 150 Candida strains to fluconazole. Results Overall correlation between microdilution and macrodilution methods was 86%. It was 91% between the Minimal Inhibitory Concentrations obtained from macrodilution and disc diffusion zone diameters. Conclusion The disc diffusion test was evaluated as a low-cost, reproducible, and efficient way of assessing the in vitro susceptibility of Candida strains to fluconazole.

  13. Inactivation of Staphylococcus aureus and Salmonella enteritidis in tryptic soy broth and caviar samples by high pressure processing

    Directory of Open Access Journals (Sweden)

    F. Fioretto

    2005-08-01

    Full Text Available We studied the action of high pressure processing on the inactivation of two foodborne pathogens, Staphylococcus aureus ATCC 6538 and Salmonella enteritidis ATCC 13076, suspended in a culture medium and inoculated into caviar samples. The baroresistance of the two pathogens in a tryptic soy broth suspension at a concentration of 10(8-10(9 colony-forming units/ml was tested for continuous and cycled pressurization in the 150- to 550-MPa range and for 15-min treatments at room temperature. The increase of cycle number permitted the reduction of the pressure level able to totally inactivate both microorganisms in the tryptic soy broth suspension, whereas the effect of different procedure times on complete inactivation of the microorganisms inoculated into caviar was similar.

  14. Inactivation of Staphylococcus aureus and Salmonella enteritidis in tryptic soy broth and caviar samples by high pressure processing.

    Science.gov (United States)

    Fioretto, F; Cruz, C; Largeteau, A; Sarli, T A; Demazeau, G; El Moueffak, A

    2005-08-01

    We studied the action of high pressure processing on the inactivation of two foodborne pathogens, Staphylococcus aureus ATCC 6538 and Salmonella enteritidis ATCC 13076, suspended in a culture medium and inoculated into caviar samples. The baroresistance of the two pathogens in a tryptic soy broth suspension at a concentration of 10(8)-10(9) colony-forming units/ml was tested for continuous and cycled pressurization in the 150- to 550-MPa range and for 15-min treatments at room temperature. The increase of cycle number permitted the reduction of the pressure level able to totally inactivate both microorganisms in the tryptic soy broth suspension, whereas the effect of different procedure times on complete inactivation of the microorganisms inoculated into caviar was similar. PMID:16082468

  15. Pharmacia and biological functionalities of nutrient broth dispersed multi-walled carbon nanotubes:A novel drug delivery system

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A new drug delivery system was developed using the interaction of nutrient broth treated multi-walled carbon nanotubes(NBT-MWCNTs) and cefotaxime sodium(CTX) as a model.Investigated factors of the drug delivery system include dispersion effect,biocompatibility of NBT-MWCNTs,pharmacodynamic effect and delivery efficiency in vitro.It was found that MWCNTs can be well dispersed in the nutrient broth and stable at least for one week at 4 °C.The formed NBT-MWCNTs suspension scarcely exhibits toxicity to E.coli at concentrations lower than 10.24 μg/mL,but displays enhanced pharmacodynamic effect of CTX via its bridge effect and targeted transport.Compared with general acid treated MWCNTs(AT-MWCNTs),our present NBT-MWCNTs show good biocompatibility,enhanced pharmacodynamic effect,and high delivery efficiency.

  16. Submerged-Culture Mycelia and Broth of the Maitake Medicinal Mushroom Grifola frondosa (Higher Basidiomycetes) Alleviate Type 2 Diabetes-Induced Alterations in Immunocytic Function.

    Science.gov (United States)

    Chen, Ya-Hui; Lee, Chien-Hsing; Hsu, Tai-Hao; Lo, Hui-Chen

    2015-01-01

    Type 2 Diabetes mellitus (T2DM), a disease with impaired glucose, protein and lipid metabolism, low-grade chronic inflammation, and immune dysfunction, is a global public health crisis. We previously demonstrated that Grifola frondosa has bioactivities in improving glycemic responses in diabetic rats. Herein, we investigated the immunomodulatory effects of the submerged-culture mycelia and broth of G. frondosa on the peripheral blood cells (PBL) and splenocytes. Male Wistar rats were administered with saline (normal rats) or streptozotocin plus nicotinamide (T2DM rats) and were intragastrically administered with placebo, fermented mycelia, broth, or mycelia plus broth (1 g kg-1 day-1) for two weeks. In normal rats, ingestion of mycelia significantly decreased monocytes and ingestion of mycelia and broth significantly decreased the productions of interferon (IFN)-γ and interleukin (IL)-4 from the PBL and splenocytes. In T2DM rats, ingestion of mycelia, broth, and mycelia plus broth significantly alleviated the increases in 2 h postprandial blood glucose and the productions of IFN-γ from the T-leukocytes, IL-4, and IL-6 from the monocytes and IL-4 from the T-splenocytes, as well as significantly improved the productions of tumor-necrosis factor-α from the macrophages. In conclusion, submerged-culture mycelia and broth of G. frondosa may decrease cell-medicated immunity in normal rats and improve hyperglycemia and diabetes-induced alterations in cell-medicated and innate immunities in T2DM rats. PMID:26349512

  17. Effect of cycle time on fungal morphology, broth rheology, and recombinant enzyme productivity during pulsed addition of limiting carbon source.

    Science.gov (United States)

    Bhargava, Swapnil; Wenger, Kevin S; Rane, Kishore; Rising, Vanessa; Marten, Mark R

    2005-03-01

    For many years, high broth viscosity has remained a key challenge in large-scale filamentous fungal fermentations. In previous studies, we showed that broth viscosity could be reduced by pulsed addition of limiting carbon during fed-batch fermentation. The objective in this study was to determine how changing the frequency of pulsed substrate addition affects fungal morphology, broth rheology, and recombinant enzyme productivity. To accomplish this, a series of duplicate fed-batch fermentations were performed in 20-L fermentors with a recombinant glucoamylase producing strain of Aspergillus oryzae. The total cycle time for substrate pulsing was varied over a wide range (30-2,700 s), with substrate added only during the first 30% of each cycle. As a control, a fermentation was conducted with continuous substrate feeding, and in all fermentations the same total amount of substrate was added. Results show that the total biomass concentration remained relatively unaltered, while a substantial decrease in the mean projected area of fungal elements (i.e., average size) was observed with increasing cycle time. This led to reduced broth viscosity and increased oxygen uptake rate. However, high values of cycle time (i.e., 900-2,700 s) showed a significant increase in fungal conidia formation and significantly reduced recombinant enzyme productivity, suggesting that the fungi channeled substrate to storage compounds rather than to recombinant protein. In addition to explaining the effect of cycle time on fermentation performance, these results may aid in explaining the discrepancies observed on scale-up to larger fermentors. PMID:15643626

  18. Development of a repair-enrichment broth for resuscitation of heat-injured Listeria monocytogenes and Listeria innocua.

    OpenAIRE

    Busch, S V; Donnelly, C W

    1992-01-01

    The ability of the divalent cations magnesium, iron, calcium and manganese; yeast extract; pyruvate; catalase; and the carbohydrates glucose, lactose, sucrose, esculin, fructose, galactose, maltose, and mannose to facilitate repair of heat-injured Listeria monocytogenes and Listeria innocua was evaluated. Listeria populations were injured by heating at 56 degrees C for 50 min. To determine the effects on repair, Trypticase soy broth (TSB) was supplemented with each medium component to be eval...

  19. Extraction of natural red colorants from the fermented broth of Penicillium purpurogenum using aqueous two-phase polymer systems.

    Science.gov (United States)

    Santos-Ebinuma, Valéria Carvalho; Lopes, André Moreni; Pessoa, Adalberto; Teixeira, Maria Francisca Simas

    2015-01-01

    Safety concerns related to the increasing and widespread application of synthetic coloring agents have increased the demand for natural colorants. Fungi have been employed in the production of novel and safer colorants. In order to obtain the colorants from fermented broth, suitable extraction systems must be developed. Aqueous two-phase polymer systems (ATPPS) offer a favorable chemical environment and provide a promising alternative for extracting and solubilizing these molecules. The aim of this study was to investigate the partitioning of red colorants from the fermented broth of Penicillium purpurogenum using an ATPPS composed of poly(ethylene glycol) (PEG) and sodium polyacrylate (NaPA). Red colorants partitioned preferentially to the top (PEG-rich phase). In systems composed of PEG 6,000 g/mol/NaPA 8,000 g/mol, optimum colorant partition coefficient (KC ) was obtained in the presence of NaCl 0.1 M (KC  = 10.30) while the PEG 10,000 g/mol/NaPA 8,000 g/mol system in the presence of Na2 SO4 0.5 M showed the highest KC (14.78). For both polymers, the mass balance (%MB) and yield in the PEG phase (%ηTOP ) were close to 100 and 79%, respectively. The protein selectivity in all conditions evaluated ranged from 2.0-3.0, which shows a suitable separation of the red colorants and proteins present in the fermented broth. The results suggest that the partitioning of the red colorants is dependent on both the PEG molecular size and salt type. Furthermore, the results obtained support the potential application of ATPPS as the first step of a purification process to recover colorants from fermented broth of microorganisms. PMID:26097197

  20. Efficacy of Plectranthus amboinicus (Lour.) Spreng in a Murine Model of Methicillin-Resistant Staphylococcus aureus Skin Abscesses

    OpenAIRE

    Francisco Fábio Martins de Oliveira; Alba Fabiola Torres; Thially Braga Gonçalves; Gilvandete Maria Pinheiro Santiago; Cibele Barreto Mano de Carvalho; Milena Braga Aguiar; Lilia Maria Carneiro Camara; Silvia Helena Rabenhorst; Alice Maria Costa Martins; José Telmo Valença Junior; Aparecida Tiemi Nagao-Dias

    2013-01-01

    The present work aimed to evaluate the effectiveness of Plectranthus amboinicus (Lour.) Spreng against MRSA clinical isolates. The in vitro antimicrobial activity of the hydroalcoholic extract (HE), the ethyl acetate (EA) fraction and its subfractions were determined by broth microdilution and bioautography against MRSA clinical isolates. The microdilution checkerboard method was used to assess in vitro drug combination studies. To induce abscess formation, bacterial suspensions were added to...

  1. The effects of fermentation and adsorption using lactic acid bacteria culture broth on the feed quality of rice straw

    Institute of Scientific and Technical Information of China (English)

    LIU Jing-jing; LIU Xiao-ping; REN Ji-wei; ZHAO Hong-yan; YUAN Xu-feng; WANG Xiao-fen; Abdelfattah Z M Salem; CUI Zong-jun

    2015-01-01

    To improve the nutritional value and the palatability of air-dried rice straw, culture broth of the lactic acid bacteria community SFC-2 was used to examine the effects of two different treatments, fermentation and adsorption. Air-dried and chopped rice straw was treated with either fermentation for 30 d after adding 1.5 L nutrient solution (50 mL inocula L–1, 1.2×1012 CFU mL–1 inocula) kg–1 straw dry matter, or spraying a large amount of culture broth (1.5 L kg–1 straw dry matter, 1.5×1011 CFU mL–1 culture broth) on the straw and al owing it to adsorb for 30 min. The feed quality and aerobic stability of the resulting forage were examined. Both treatments improved the feed quality of rice straw, and adsorption was better than fermen-tation for preserving nutrients and improving digestibility, as evidenced by higher dry matter (DM) and crude protein (CP) concentrations, lower neutral detergent ifber (NDF), acid detergent ifber (ADF) and NH3-N concentrations, as wel as higher lactic acid production and in vitro digestibility of DM (IVDMD). The aerobic stability of the adsorbed straw and the fermented straw was 392 and 480 h, respectively. After being exposed to air, chemical components and microbial community of the fermented straw were more stable than the adsorbed straw.

  2. Post-irradiation modification of radiation-induced heteroallelic reversion in diploid yeast: effect of nutrient broth

    International Nuclear Information System (INIS)

    The effect of post-irradiation growth in complete rich medium on the expression of the reversion to arginine-independence induced by gamma and alpha radiation in a heteroallelic diploid yeast strain (Saccharomyces cerevisiae BZ34) has been studied. During the post-irradiation treatment the reversion frequency increased, reached a peak at about 90 minutes and decreased thereafter reaching a constant value for treatment periods exceeding 6h. As determined by the increase in number of budding cells, extensive DNA synthesis took place in cells incubated only in the nutrient medium and not in the omission medium. Hence the observed increase in the reversion frequency is explained on the basis that post-irradiation DNA synthesis is necessary for the expression of gene conversion. The decrease in the reversion frequency for continued treatment with yeast extract, peptone, dextrose (YEPD) is related to the fact that only one daughter of the post-irradiation first cell division is a revertant. The broth effect was not lost when the irradiated cells were first incubated for 90 min in arginine-less medium and then transferred to the broth. Similarly, the broth effect even persisted at doses high enough to induce considerable division delay. These results suggest that the radiation-induced pre-conversional lesions are not susceptible to repair by alternative pathways

  3. Combination of Origanum vulgare L. essential oil and lactic acid to inhibit Staphylococcus aureus in meat broth and meat model

    Directory of Open Access Journals (Sweden)

    Jefferson C. de Barros

    2012-09-01

    Full Text Available This study assessed the occurrence of an enhancing inhibitory effect of the combined application of Origanum vulgare L. essential oil and lactic acid against Staphylococcus aureus by the determination of Fractional Inhibitory Concentration (FIC index and cell viability in meat broth and meat model. Minimum Inhibitory Concentration (MIC and Minimum Bactericidal Concentration (MBC of the oil was 0.6 and 1.25 µL.mL-1, respectively. Lactic acid showed MIC and MBC of 2.5 and 5µL.mL-1, respectively. FIC indices of the combined application of the oil and lactic acid were 0.5 showing a synergic interaction. The essential oil and lactic acid showed similar (p>0.05 anti-S. aureus effect in meat broth over 96 h of exposure. Treatment with essential oil or lactic acid presented a smaller anti-staphylococcal effect in meat in comparison to meat broth. No significant difference (p>0.05 was found for the microbial counts in meat treated with each antimicrobial alone or in mixture. These results could arise as an interesting approach for the improvement of food preservation using more natural procedures, considering the current demand of consumer and sensory quality of foods.

  4. Modeling the behavior of Listeria monocytogenes during enrichment in half Fraser broth; impact of pooling and the duration of enrichment on the detection of L. monocytogenes in food.

    Science.gov (United States)

    Augustin, Jean-Christophe; Kalmokoff, Martin; Ells, Timothy; Favret, Sandra; Desreumaux, Jennifer; Decourseulles Brasseur, Emilie; Gnanou Besse, Nathalie

    2016-12-01

    A stochastic model describing the growth of Listeria monocytogenes during enrichment in half Fraser was developed for the purpose of estimating the effects of modifications to the first enrichment step of the EN ISO 11290-1 detection method. Information pertaining to the variability of growth rates, physiological state of the cell, and the behavior of individual cells contaminating the food were obtained from previously published studies. We used this model to investigate the impact of pooling enrichment broths (wet pooling) on the performance of the standard method. For validation of the model, the numbers of L. monocytogenes occurring in 88 naturally contaminated foods following pre-enrichment were compared to model-simulated microbial counts. The model was then used to perform simulations representative of the natural contamination observed for smoked salmon in the European baseline survey of 2010-2011. The model-estimated L. monocytogenes levels following individual enrichment or following the pooling of five broths where only one would be contaminated were compared. The model indicated a 10% loss of method sensitivity resulting from wet pooling. The model also predicted a 5% decrease in the sensitivity of the method when the duration of the enrichment was reduced from 24 to 22 h. PMID:27554154

  5. Comparison of disc and MIC reduction methods with polymerase chain reaction for the detection of metallo-β-lactamase in Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    S Buchunde

    2012-01-01

    Full Text Available Purpose: The present study was undertaken to evaluate the screening antibiotic, confirmatory phenotypic test and agent against PCR as gold standard and to detect the prevalent MBL gene. Materials and Methods: Three hundred and twenty-six Pseudomonas aeruginosa isolates were screened for resistance to Imipenem (IPM, Meropemem (MEM and Ceftazidime (CAZ by disc diffusion. Isolates resistant to any of these were considered screen test-positive for MBL and were subjected to Double disc synergy test (DDST and Disc potentiation test (DPT: Using IPM, MEM and CAZ alone and with EDTA, Minimum inhibitory concentration (MIC reduction [four-fold or more reduction in MIC of IPM and MEM in presence of chelators: EDTA and 1,10-phenanthroline (EPI/EPM: EDTA-phenanthroline- Imipenem/Meropenem Broth Microdilution method] and polymerase chain reaction (PCR for blaIMP and blaVIM . Results: Screen test-positives by MEM and CAZ were 19.3% as against 17.8% by IPM. MEMDDST, DPT and EPM confirmed 100% screen-test positives as against 93.7% by CAZ DDST and DPT-2, 76.2% by CAZ DPT-1, 88.9% by IPM DDST, 85.7% by IPM DPT-1 and 92.1% by EPI. IPMand CAZ DDST together confirmed 100% while IPM and CAZ DPT-2 confirmed 96.8%. All 63 screen-test positives showed the presence of blaVIM . Conclusions: MEM was found to be the best screening and confirmatory agent for MBL detection and blaVIM was found to be the prevalent MBL gene in this part of the country.

  6. Biofilm formation by Staphylococcus aureus from food contact surfaces in a meat-based broth and sensitivity to sanitizers

    OpenAIRE

    Evandro Leite de Souza; Quênia Gramile Silva Meira; Isabella de Medeiros Barbosa; Ana Júlia Alves Aguiar Athayde; Maria Lúcia da Conceição; José Pinto de Siqueira Júnior

    2014-01-01

    This study assessed the capacity of adhesion, the detachment kinetic and the biofilm formation by Staphylococcus aureus isolated from food services on stainless steel and polypropylene surfaces (2 × 2 cm) when cultivated in a meat-based broth at 28 and 7 °C. It was also to study the efficacy of the sanitizers sodium hypochlorite (250 mg/L) and peracetic acid (30 mg/L) in inactivating the bacterial cells in the preformed biofilm. S. aureus strains adhered in high numbers regardless the assayed...

  7. Two new cyclopeptides from the co-culture broth of two marine mangrove fungi and their antifungal activity

    OpenAIRE

    Song Huang; Weijia Ding; Chunyuan Li; Cox, Daniel G.

    2014-01-01

    Background: The strategy that co-cultivation two microorganisms in a single confined environment were recently developed to generate new active natural products. In the study, two new cyclic tetrapeptides, cyclo (D-Pro-L-Tyr-L-Pro-L-Tyr) (1) and cyclo (Gly-L-Phe-L-Pro-L-Tyr) (2) were isolated from the co-culture broth of two mangrove fungi Phomopsis sp. K38 and Alternaria sp. E33. Their antifungal activity against Candida albicans, Gaeumannomyces graminis, Rhzioctonia cerealis, Helminthospori...

  8. The influence of cholesterol and biomass concentration on the uptake of cholesterol by Lactobacillus from MRS broth

    Directory of Open Access Journals (Sweden)

    Małgorzata Ziarno

    2007-06-01

    Full Text Available The aim of this study was the determination of some factors influence (i.e. the vitality of bacteria cells and the cholesterol concentration on the ability of selected Lactobacillus sp. to cholesterol uptake during culture in MRS broth. Three Lactobacillus strains (Lb. delbrueckii subsp. bulgaricus, Lb. acidophilus, Lb. casei isolated from commercial single species lyophilized dairy starter cultures and three Lactobacillus strains (Lb. plantarum, Lb. delbrueckii subsp. bulgaricus, Lb. acidophilus originated from commercial pharmaceuticals were used in this study. The uptake of cholesterol from MRS broth during the growth of Lactobacillus sp., expressed as the difference between the final and the initial concentrations of cholesterol, ranged from 0.053 to 0.153 g/dm³, apart from the initial cholesterol content and the origin of Lactobacillus sp. The results confirmed that biomass concentration have a statistically significant effect on uptake of cholesterol. The ten-fold increase of the amount of intact cells biomass caused about 1.5-2-fold increase of the amount of cholesterol removed. The influence of the concentration of biomass of alive cells on the removal of cholesterol was bigger than in case of the heat-sterilized cells.

  9. HPLC-fluorescence detection method for determination of key intermediates of the lincomycin biosynthesis in fermentation broth

    Czech Academy of Sciences Publication Activity Database

    Kameník, Zdeněk; Kopecký, J.; Marečková, M.; Ulanová, Dana; Novotná, Jitka; Pospíšil, Stanislav; Olšovská, Jana

    2009-01-01

    Roč. 393, 6-7 (2009), s. 1779-1787. ISSN 1618-2642 R&D Projects: GA ČR GA204/05/0616; GA AV ČR IAA6020410; GA MŠk 2B08064 Institutional research plan: CEZ:AV0Z50200510 Keywords : Lincomycin precursors * o-Phthaldialdehyde * Fluorescence detection Subject RIV: CB - Analytical Chemistry, Separation Impact factor: 3.480, year: 2009

  10. Comparação da eficiência dos caldos de enriquecimento seletivo no isolamento de Salmonella Dublin Comparison of the efficiency of selective enrichment broths for Salmonella Dublin isolation

    Directory of Open Access Journals (Sweden)

    D.G. Silva

    2008-06-01

    Full Text Available The aim of this study was to compare three different selective enrichment broths: Rappaport-Vassiliadis (RV, selenite cystine (SC and Muller-Kauffmann tetrathionate (MKT for Salmonella Dublin isolation from faecal samples of calf experimentally infected. The bacteriological procedure involved pre-enrichment stages in Hajna-GN broth (only for the samples inoculated in RV broth, selective enrichment, culture in modified brilliant green agar (BGA, presumptive biochemistry tests (using triple-sugar-iron agar and lysine-agar and slide agglutination test with poli-O and poli-H Salmonella antiserum. The effects of enrichment temperatures using RV broth were also evaluated (37ºC and 42ºC. SC broth was significantly more efficient in the isolation of Salmonella Dublin (P<0,05, whereas RV broth incubated at 42ºC had a lower efficiency in the microbiological isolation.

  11. Comparação da eficiência dos caldos de enriquecimento seletivo no isolamento de Salmonella Dublin Comparison of the efficiency of selective enrichment broths for Salmonella Dublin isolation

    OpenAIRE

    D.G. Silva; J.J. Fagliari; T.B. Garcia

    2008-01-01

    The aim of this study was to compare three different selective enrichment broths: Rappaport-Vassiliadis (RV), selenite cystine (SC) and Muller-Kauffmann tetrathionate (MKT) for Salmonella Dublin isolation from faecal samples of calf experimentally infected. The bacteriological procedure involved pre-enrichment stages in Hajna-GN broth (only for the samples inoculated in RV broth), selective enrichment, culture in modified brilliant green agar (BGA), presumptive biochemistry tests (using tripl...

  12. Antimicrobial activity of Ethiopian medicinal plants

    OpenAIRE

    Bernášková, Eva

    2013-01-01

    In vitro antimicrobial activity of eighteen Ethiopian medicinal plant species that were selected based on ethnobotanical information on their traditional use to treat infectious diseases was determined by the broth microdilution method. The antimicrobial activity of ethanol extracts of selected plants against potentially pathogenic microorganism such as Bacillus cereus, Bacteroides fragilis, Candida albicans, Clostridium perfringens, Enterococcus faecalis, Escherichia coli, Listeria monocytog...

  13. [Comparative study, using 3 methods, of the sensitivity to metronidazole and ornidazole of anaerobic or related bacteria].

    Science.gov (United States)

    Gallusser, A

    1983-01-01

    A comparative study of the sensitivity to metronidazole and ornidazole of 127 anaerobic or microaerophilic strains isolated from various clinical samples showed that the activity of both products was similar: the distribution of sensitive and resistant strains was identical. However, the in vitro activity level of metronidazole was slightly higher. This difference, though statistically significant, had no incidence on therapeutic indications. The determination of sensitivity towards the two nitroimidazoles was carried out by three methods: broth dilution and agar diffusion for metronidazole; and broth dilution and disk-broth for ornidazole. Two of these methods, broth dilution and disk-broth, gave concordant results. Conversely, the limits of the agar diffusion technique were shown to be related to independent biological factors such as bacterial motility and slow growth rate. The poor accuracy of this method limits its use in detecting total resistance. PMID:6651124

  14. Detection of Burkholderia pseudomallei in Sputum using Selective Enrichment Broth and Ashdown’s Medium at Kampong Cham Provincial Hospital, Cambodia [v1; ref status: indexed, http://f1000r.es/4w7

    Directory of Open Access Journals (Sweden)

    Somary Nhem

    2014-12-01

    Full Text Available Melioidosis infection, caused by Burkholderia pseudomallei, is increasingly reported in Cambodia. We hypothesized that implementation of an enhanced sputum testing protocol in a provincial hospital diagnostic microbiology laboratory would increase detection of B. pseudomallei. We tested 241 sputum specimens that were deemed acceptable for culture, comparing culture in selective enrichment broth followed by sub-culture on Ashdown’s medium to standard culture methods. Two specimens (0.8% were positive for B. pseudomallei using the enhanced protocol whereas one specimen (0.4% was positive using standard methods. These findings demonstrate that B. pseudomallei is rarely detected in sputum at this hospital. The low frequency of B. pseudomallei in sputum specimens precludes drawing any conclusions about the relative benefits of an enhanced sputum testing protocol at this site. Promoting clinician awareness of the infection and encouraging utilization of diagnostic microbiology services are likely to be important factors in facilitating identification of melioidosis.

  15. Simultaneous determination of colistin and levofloxacin in broth by LC-MS/MS%LC-MS/MS法同时测定多粘菌素E和左氧氟沙星的浓度

    Institute of Scientific and Technical Information of China (English)

    梅和坤; 王睿; 白楠; 梁蓓蓓; 曹江; 汶柯; 唐铭婧; 刘银萍; 李悦

    2013-01-01

    Objective To establish a HPLC - MS/MS method for determining colistin and levofloxaein in broth simultaneously. Methods After the sample pretreated with solid - phase extraction ( SPE) , colistin and levofloxaein were separated on a ZORBAX SB - C18 analytical column by using the mobile phase of acetonitrile(0. 1% formic acid) - water(0. 1% formic acid) (50'- 50) at a flow rate of 0. 3 mL ? min -1'. Results The linear range of colistin and levofloxaein in broth was 0. 48 -9. 60μg ? mL-1. Lower limit of quantification was 0. 48 μg ? mL-1 and precision of intra - day( RSD) was ≤6. 83% , precision of inter - day ( RSD) was ≤9. 85% . Conclusion The method is sensitive, fast and accurate, which is suitable for determination of colistin and levofloxaein in broth.%目的 建立同步测定培养液中多粘菌素E和左氧氟沙星的LC-MS/MS方法.方法 待测样品用固相萃取(SPE)法预处理后,色谱柱为ZORBAXSB-C18柱,流动相为乙腈(0.1%甲酸)-水(0.1%甲酸)(50∶ 50),流速为0.3mL·min-1,用电喷雾离子源(ESI),正离子方式检测,多重反应监测(MRM)扫描.结果 多粘菌素E主要成分多粘菌素A,B和左氧氟沙星的线性范围为0.48 ~9.60 μg·mL-,定量下限为0.48 μg·mL-1,日内精密度(RSD)≤6.83%,日间RSD≤9.85%.绝对回收率为88.06% ~ 104.43%·RSD≤4.15%.结论 本方法专属性好、灵敏、准确、快速,满足了测定的要求.

  16. Suppression of pecan and peach pathogens using metabolites or broths of from symbiotic bacteria obtained from the guts of entomopathogenic nematodes

    Science.gov (United States)

    Concentrated metabolites from the bacteria Xenorhabdus spp. and Photorhabdus spp. have previously been shown to suppress growth of peach and pecan pathogens in vitro, and reduce disease on detached leaves or terminals. The objectives of this study were 1) determine if bacterial broths (in addition t...

  17. Investigations on interfaciale activity of a biosurfactant from a xanthan broth; Untersuchungen zur Grenzflaechenaktivitaet eines Biotensids aus Xanthan-Bruehen

    Energy Technology Data Exchange (ETDEWEB)

    Birdi, K.S. [Royal Danish School of Pharmacy, Kopenhagen (Denmark); Kleinitz, W. [Preussag Energie GmbH, Lingen (Germany); Littmann, W. [Littmann Consult, Wunstorf (Germany)

    1997-10-01

    In order to improve crude oil production, various field projects have been conducted by Preussag Energie GmbH. In these tests, the viscosity of the reservoir water was increased to the value of crude oil by polymer addition. It is well known that this displacement process results in enhanced oil recovery. A surprising discovery was the pressure of stable oil-in-water emulsions when xanthane a biopolymer which is stable in salt water was used in the form of the fermentation broth and if xanthane was also detected in the reservoir water in the production fluid. Industrial xanthanes from various manufacturers exhibited interfacial activity only when used as fermentation broths. Powdered products were inert. The application of fluorescence spectrometry has proved that the biosurfactant is not a protein. The interfacial activity has been determined by monolayer measurements at various pll-values and salt concentrations. The critical micelle concentration of about 100 mg/l is stable over a wide range of temperatures and salinities. Potential applications of this surfactant include the stimulation of oil and gas wells. (orig.) [Deutsch] Zur Verbesserung der Rohoelfoerderung wurden von der Preussag Energie GmbH verschiedene Feldprojekte durchgefuehrt, bei denen durch Polymerzugabe die Viskositaet des Lagerstaettenwassers auf den Betrag des Rohoels angehoben wurde. Dieser Verdraengungsvorgang fuehrt bekanntlich zu einer Erhoehung der Erdoelausbeute. Bei Einsatz des salzwasserstabilen Biopolymers Xanthan in Form der Fermentationsbruehe wurden ueberraschenderweise dann stabile Oel-in-Wasser-Emulsionen festgestellt, wenn in dem Fluid an der Produktionsbohrung auch Xanthan im Lagerstaettenwasser nachgewiesen werden konnte. Industrielle Xanthane von verschiedenen Herstellern zeigten nur bei Verwendung von Fermentationsbruehen Grenzflaechenaktivitaet. Pulverprodukte waren inert. Mittels Fluoreszenzspektrometrie konnte nachgewiesen werden dass es sich bei dem Biotensid nicht um

  18. Fast quantitative determination of microbial rhamnolipids from cultivation broths by ATR-FTIR Spectroscopy

    Directory of Open Access Journals (Sweden)

    Hausmann Rudolf

    2008-10-01

    Full Text Available Abstract Background Vibrational spectroscopic techniques are becoming increasingly important and popular because they have the potential to provide rapid and convenient solutions to routine analytical problems. Using these techniques, a variety of substances can be characterized, identified and also quantified rapidly. Results The rapid ATR-FTIR (Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy in time technique has been applied, which is suitable to quantify the concentrations of microbial rhamnolipids in a typical cultivation process. While the usually applied HPLC analysis requires an extensive and time consuming multi step extraction protocol for sample preparation, the ATR-FTIR-method allows the quantification of the rhamnolipids within 20 minutes. Accuracies between 0.5 g/l – 2.1 g/l for the different analytes were determined by cross validation of the calibration set. Even better accuracies between 0.28 g/l – 0.59 g/l were found for independent test samples of an arbitrarily selected cultivation. Conclusion ATR-FTIR was found to be suitable for the rapid analysis of rhamnolipids in a biotechnological process with good reproducibility in sample determination and sufficient accuracy. An improvement in accuracy through continuous expansion and validation of the reference spectra set seems very likely.

  19. Fabrication and characterization of silver nanoparticles using Delonix elata leaf broth

    Science.gov (United States)

    Sathiya, C. K.; Akilandeswari, S.

    2014-07-01

    The synthesis of nanoparticles from plant sources has proved to be an effective and alternative method for the novel production of nanoparticles. This paper reports the bioreduction of silver nitrate into silver nanoparticle by the leaf extract of Delonix elata. The synthesized silver nanoparticles were characterized by UV-visible (UV-vis) spectroscopy, Fourier infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Scanning Electron Microscopy (SEM) coupled with Energy Dispersive Spectroscopy (EDS), high resolution transmission electron microscope (HRTEM). In addition the size of the NPs was calculated by using Malvern Zetasizer and the stability by zeta potential. UV-vis spectra show the surface plasmon resonance (SPR) at 432 nm. This reveals the reduction of silver ions (Ag+) into silver (Ag°) and indicating the formation of silver nanoparticles (AgNPs). SEM analysis revealed the spherical shape of the particles with sizes in the range of 35-45 nm and EDS spectrum confirmed the presence of silver along with other elements in the plant metabolite. The XRD analysis showed that the AgNPs are crystalline in nature and have face-centered cubic structure. FT-IR spectra show the existence of biomolecules responsible for the reduction of silver nitrate. The size of the AgNPs estimated from particle size distribution curve shows the 70 nm. The zeta potential of AgNPs was found to be -18 mV, indicating the dispersion and stability.

  20. Efek Penambahan Glukosa pada Saburoud Dextrose Broth terhadap Pertumbuhan Candida albicans (Uji In Vitro

    Directory of Open Access Journals (Sweden)

    Lakshmi A. Leepel

    2012-10-01

    Full Text Available High carbohydrate intake is one of predisposing factors of oral candidiasis. Objective: Investigating the effect of 1%,5%,10% glucose addition on the growth of C.albicans in vitro. Method: C.albicans sample was taken from oral swab of a male oral candidiasis patient. Identification of C.albicans was conducted using CHROMagar and confirmed by germ tube formation in serum. As a comparison, C.albicans ATCC10231 was used. After 2 days the cultures were serially diluted and inoculated in SDB without glucose, and with 1%,5%,10% addditional glucose, kept for 3 and 7 days in room temperature, then inoculated in SDA. The CFU/ml were counted after 2 days. ANOVA with α0.05 was used. Result: Statisticaly, additional 1% glucose for 3 days lead to significant decreased of growth of both clinical strain and ATCC 10231 C. albicans. However, only additional 5% and 10% glucose in clinical isolate for 7 days increased the growth of C.albicans significantly. Conclusion: The effect of additional glucose on the increased growth of C.albicans in vitro is influenced by the concentration, exposure duration of glucose, and by the strain of C.albicans.DOI: 10.14693/jdi.v16i1.14

  1. Surface Activity of Sulfactin Recovered and Purified from Fermentation Broth Using a Two-Step Ultrafiltration (UF) Process

    International Nuclear Information System (INIS)

    B. subtilis under certain types of media and fermentation conditions can produce surfactant, a bio surfactant which belongs to the lipo peptide class. Surfactant has exceptional surfactant activity, and exhibits some interesting biological characteristics such as antibacterial activity, anti tumoral activity against ascites carcinoma cells, and a hypercholesterolaemia activity that inhibits cAMP phosphodiesterase, as well as having anti-HIV properties. A cost effective recovery and purification of surfactant from fermentation broth using a two-step ultrafiltration (UF) process has been developed in order to reduce the cost of surfactant production. In this study, competitive adsorption of surfactant and proteins at the air-water interface was studied using surface pressure measurements. Small volumes of bovine serum albumin (BSA) and β-casein solutions were added to the air-water interface on a Langmuir trough and allowed to stabilise before the addition of surfactant to the sub phase. Contrasting interfacial behaviour of proteins was observed with β-casein showing faster initial adsorption compared to BSA. On introduction of surfactant both proteins were displaced but a longer time were taken to displace β-casein. Overall the results showed surfactant were highly surface-active by forming a β-sheet structure at the air-water interface after reaching its critical micelle concentration (CMC) and were effective in removing both protein films, which can be explained following the orogenic mechanism. Results showed that the two-step UF process was effective to achieve high purity and fully functional surfactant. (author)

  2. Biofilm formation by Staphylococcus aureus from food contact surfaces in a meat-based broth and sensitivity to sanitizers

    Directory of Open Access Journals (Sweden)

    Evandro Leite de Souza

    2014-01-01

    Full Text Available This study assessed the capacity of adhesion, the detachment kinetic and the biofilm formation by Staphylococcus aureus isolated from food services on stainless steel and polypropylene surfaces (2 x 2 cm when cultivated in a meat-based broth at 28 and 7 ºC. It was also to study the efficacy of the sanitizers sodium hypochlorite (250 mg/L and peracetic acid (30 mg/L in inactivating the bacterial cells in the preformed biofilm. S. aureus strains adhered in high numbers regardless the assayed surface kind and incubation temperature over 72 h. Cells detachment of surfaces revealed high persistence over the incubation period. Number of cells needed for biofilm formation was noted at all experimental systems already after 3 days. Peracetic acid and sodium hypochlorite were not efficient in completely removing the cells of S. aureus adhered on polypropylene and stainless steel surfaces. From these results, the assayed strains revealed high capacity to adhere and form biofilm on polypropylene and stainless steel surfaces under different growth conditions. Moreover, the cells in biofilm matrix were resistant for total removal when submitted to the exposure to sanitizers.

  3. Biofilm formation by Staphylococcus aureus from food contact surfaces in a meat-based broth and sensitivity to sanitizers.

    Science.gov (United States)

    de Souza, Evandro Leite; Meira, Quênia Gramile Silva; de Medeiros Barbosa, Isabella; Athayde, Ana Júlia Alves Aguiar; da Conceição, Maria Lúcia; de Siqueira Júnior, José Pinto

    2014-01-01

    This study assessed the capacity of adhesion, the detachment kinetic and the biofilm formation by Staphylococcus aureus isolated from food services on stainless steel and polypropylene surfaces (2 × 2 cm) when cultivated in a meat-based broth at 28 and 7 °C. It was also to study the efficacy of the sanitizers sodium hypochlorite (250 mg/L) and peracetic acid (30 mg/L) in inactivating the bacterial cells in the preformed biofilm. S. aureus strains adhered in high numbers regardless the assayed surface kind and incubation temperature over 72 h. Cells detachment of surfaces revealed high persistence over the incubation period. Number of cells needed for biofilm formation was noted at all experimental systems already after 3 days. Peracetic acid and sodium hypochlorite were not efficient in completely removing the cells of S. aureus adhered on polypropylene and stainless steel surfaces. From these results, the assayed strains revealed high capacity to adhere and form biofilm on polypropylene and stainless steel surfaces under different growth conditions. Moreover, the cells in biofilm matrix were resistant for total removal when submitted to the exposure to sanitizers. PMID:24948915

  4. Fate of Alicyclobacillus spp. in enrichment broth and in juice concentrates.

    Science.gov (United States)

    Oteiza, Juan Martin; Soto, Silvina; Alvarenga, Verônica O; Sant'Ana, Anderson S; Gianuzzi, Leda

    2015-10-01

    In this study, the fate of Alicyclobacillus acidoterrestris spores in different types of juice concentrates stored under different conditions was investigated. In addition, the impact of dilution procedures during the enrichment step for the detection of Alicyclobacillus in lemon juice concentrates was studied. Pear, red grape, mango, tangerine, carrot and lemon juice concentrates (50-69.4°Brix, pH1.7-4.3) were inoculated with A. acidoterrestris spores (10(3) spore/mL) and stored at 4 °C and 20 °C, after which the spores were counted at 0, 2, 5, 9, 17, 21, 28, 36, 43, and 50 days. No significant differences in the number of Alicyclobacillus spores were observed at storage temperatures of 4 °C and 20 °C (p>0.05). The results also indicated that the number of spores of A. acidoterrestris remained stable in all types of juice concentrates during the storage period, except in lemon juice concentrate. In lemon juice concentrate, a decline in A. acidoterrestris spore populations of 0.3-0.8 log CFU/mL was observed within 5-10 days of storage. The decline in A. acidoterrestris spore populations was more pronounced in cloudy lemon juice concentrate, which contained higher concentrations of flavonoids (mainly eriocitrin and hesperidin) than clarified lemon juice concentrate. It was also found that dilution of lemon juice concentrate samples in the proportion of 1:19 allowed the germination of A. acidoterrestris spores and the growth of populations of up to 10(7) CFU/mL. In contrast, the proportion (1:9) recommended in internationally recognized methods led to a reduction in the population of this microorganism that would yield false negative results. Data presented in this study demonstrated that Alicyclobacillus spores remain stable in most juice concentrates during storage, but that natural antimicrobial compounds present in some of them may decrease spore counts and inhibit their recovery by detection procedures. PMID:26102554

  5. 蜜环菌发酵液中5-羟甲基糠醛提取工艺优化%Optimization of Extracting Process of 5-Hydroxymethylfurfural from Armillaria mellea Fermentation Broth

    Institute of Scientific and Technical Information of China (English)

    陈楠; 焦连庆; 郑毅男; 于敏; 刘晓杰

    2012-01-01

    以蜜环菌发酵液为原料,通过正交试验优选5-羟甲基糠醛的提取工艺.超声提取蜜环菌发酵液中5-羟甲基糠醛,运用高效液相色谱法测定其含量,并用L9(33)正交试验设计,考察提取溶剂量(mLg)、提取时间、提取次数对提取率的影响.蜜环菌发酵液中5-羟甲基糠醛的最佳提取条件为:提取溶剂量6倍量、提取时间20min、提取次数2次.在最佳提取条件下,5-羟甲基糠醛提取率为6.12%.%The orthogonal experimental design method was used to optimize the extracting process of 5-Hydroxymethylfurfural (5-HMF) in the sample of A rmillaria mellea fermentation broth. 5-HMF was extracted by supersonic wave from Armillaria mellea fermentation broth. Then the content of 5-HMF was determined by high performance liquid chromatography. The L, (3s) orthogonal experimental design was used to study the influence of different dosage of extraction solvent (mL: g), extraction time, and the number of extraction on the extraction rate of 5-HMF. The optimum extraction condition: the dosage of extraction solvent was 6 times, extraction time was 20 min, and the number of extraction times was 2. 5-HMF's extraction rate was 6.12% in the best extraction conditions.

  6. Chemical Analysis of a "Miller-Type" Complex Prebiotic Broth : Part I: Chemical Diversity, Oxygen and Nitrogen Based Polymers.

    Science.gov (United States)

    Wollrab, Eva; Scherer, Sabrina; Aubriet, Frédéric; Carré, Vincent; Carlomagno, Teresa; Codutti, Luca; Ott, Albrecht

    2016-06-01

    In a famous experiment Stanley Miller showed that a large number of organic substances can emerge from sparking a mixture of methane, ammonia and hydrogen in the presence of water (Miller, Science 117:528-529, 1953). Among these substances Miller identified different amino acids, and he concluded that prebiotic events may well have produced many of Life's molecular building blocks. There have been many variants of the original experiment since, including different gas mixtures (Miller, J Am Chem Soc 77:2351-2361, 1955; Oró Nature 197:862-867, 1963; Schlesinger and Miller, J Mol Evol 19:376-382, 1983; Miyakawa et al., Proc Natl Acad Sci 99:14,628-14,631, 2002). Recently some of Miller's remaining original samples were analyzed with modern equipment (Johnson et al. Science 322:404-404, 2008; Parker et al. Proc Natl Acad Sci 108:5526-5531, 2011) and a total of 23 racemic amino acids were identified. To give an overview of the chemical variety of a possible prebiotic broth, here we analyze a "Miller type" experiment using state of the art mass spectrometry and NMR spectroscopy. We identify substances of a wide range of saturation, which can be hydrophilic, hydrophobic or amphiphilic in nature. Often the molecules contain heteroatoms, with amines and amides being prominent classes of molecule. In some samples we detect ethylene glycol based polymers. Their formation in water requires the presence of a catalyst. Contrary to expectations, we cannot identify any preferred reaction product. The capacity to spontaneously produce this extremely high degree of molecular variety in a very simple experiment is a remarkable feature of organic chemistry and possibly prerequisite for Life to emerge. It remains a future task to uncover how dedicated, organized chemical reaction pathways may have arisen from this degree of complexity. PMID:26508401

  7. Partial purification of saccharifying and cell wall-hydrolyzing enzymes from malt in waste from beer fermentation broth.

    Science.gov (United States)

    Khattak, Waleed Ahmad; Kang, Minkyung; Ul-Islam, Mazhar; Park, Joong Kon

    2013-06-01

    A number of hydrolyzing enzymes that are secreted from malt during brewing, including cell wall-hydrolyzing, saccharide-hydrolyzing, protein-degrading, lipid-hydrolyzing, and polyphenol and thiol-hydrolyzing enzymes, are expected to exist in an active form in waste from beer fermentation broth (WBFB). In this study, the existence of these enzymes was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis, after which enzyme extract was partially purified through a series of purification steps. The hydrolyzing enzyme activity was then measured under various conditions at each purification step using carboxymethyl cellulose as a substrate. The best hydrolyzing activities of partially purified enzymes were found at pH 4.5 and 50 °C in a citrate buffer system. The enzymes showed highest thermal stability at 30 °C when exposed for prolonged time. As the temperature increased gradually from 25 to 70 °C, yeast cells in the chemically defined medium with enzyme extract lost their cell wall and viability earlier than those without enzyme extract. Cell wall degradation and the release of cell matrix into the culture media at elevated temperature (45-70 °C) in the presence of enzyme extract were monitored through microscopic pictures. Saccharification enzymes from malt were relatively more active in the original WBFB than supernatant and diluted sediments. The presence of hydrolyzing enzymes from malt in WBFB is expected to play a role in bioethanol production using simultaneous saccharification and fermentation without the need for additional enzymes, nutrients, or microbial cells via a cell-free enzyme system. PMID:23377262

  8. Chemical Analysis of a "Miller-Type" Complex Prebiotic Broth. Part I: Chemical Diversity, Oxygen and Nitrogen Based Polymers

    Science.gov (United States)

    Wollrab, Eva; Scherer, Sabrina; Aubriet, Frédéric; Carré, Vincent; Carlomagno, Teresa; Codutti, Luca; Ott, Albrecht

    2016-06-01

    In a famous experiment Stanley Miller showed that a large number of organic substances can emerge from sparking a mixture of methane, ammonia and hydrogen in the presence of water (Miller, Science 117:528-529, 1953). Among these substances Miller identified different amino acids, and he concluded that prebiotic events may well have produced many of Life's molecular building blocks. There have been many variants of the original experiment since, including different gas mixtures (Miller, J Am Chem Soc 77:2351-2361, 1955; Oró Nature 197:862-867, 1963; Schlesinger and Miller, J Mol Evol 19:376-382, 1983; Miyakawa et al., Proc Natl Acad Sci 99:14,628-14,631, 2002). Recently some of Miller's remaining original samples were analyzed with modern equipment (Johnson et al. Science 322:404-404, 2008; Parker et al. Proc Natl Acad Sci 108:5526-5531, 2011) and a total of 23 racemic amino acids were identified. To give an overview of the chemical variety of a possible prebiotic broth, here we analyze a "Miller type" experiment using state of the art mass spectrometry and NMR spectroscopy. We identify substances of a wide range of saturation, which can be hydrophilic, hydrophobic or amphiphilic in nature. Often the molecules contain heteroatoms, with amines and amides being prominent classes of molecule. In some samples we detect ethylene glycol based polymers. Their formation in water requires the presence of a catalyst. Contrary to expectations, we cannot identify any preferred reaction product. The capacity to spontaneously produce this extremely high degree of molecular variety in a very simple experiment is a remarkable feature of organic chemistry and possibly prerequisite for Life to emerge. It remains a future task to uncover how dedicated, organized chemical reaction pathways may have arisen from this degree of complexity.

  9. Hypolipidemic Effects of Biopolymers Extracted from Culture Broth, Mycelia, and Fruiting Bodies of Auricularia auricula-judae in Dietary-induced Hyperlipidemic Rats

    OpenAIRE

    Jeong, Hun; Yang, Byung-Keun; Jeong, Yong-Tae; Kim, Guk-Nam; Jeong, Yu-Sun; Kim, Sang-Min; Mehta, Pradeep; Song, Chi-Hyun

    2007-01-01

    Hypolipidemic effect of biopolymers extracted from culture broth (CP), mycelia (MP), and fruiting bodies (FP) of Auricularia auricula-judae was investigated in dietary-induced hyperlipidemic rats. The experimental animals were administrated (100 mg/kg body weight) with different biopolymers, daily for 4 weeks. Hypolipidemic effects were achieved in all the experimental groups, however, FP was proved to be the most potent one. The administration of the FP reduced the plasma triglyceride, total...

  10. Determination of Amino Acids in Cell Culture and Fermentation Broth Media Using Anion-Exchange Chromatography with Integrated Pulsed Amperometric Detection

    OpenAIRE

    Hanko, Valoran P.; Heckenberg, Andrea; Rohrer, Jeffrey S.

    2004-01-01

    Anion-exchange chromatography with integrated pulsed amperometric detection (AE-IPAD) separates and directly detects amino acids, carbohydrates, alditols, and glycols in the same injection without pre- or post-column derivatization. These separations use a combination of NaOH and NaOH/sodium acetate eluents. We previously published the successful use of this technique, also known as AAA-Direct, to determine free amino acids in cell culture and fermentation broth media. We showed that retentio...

  11. Presumptive identification of Clostridium difficile by detection of p-cresol in prepared peptone yeast glucose broth supplemented with p-hydroxyphenylacetic acid.

    OpenAIRE

    Sivsammye, G; Sims, H V

    1990-01-01

    Prereduced, anaerobically sterilized peptone yeast glucose broth was supplemented with p-hydroxyphenylacetic acid and used for the presumptive identification of Clostridium difficile. Two hundred eighty-two organisms were grown in this medium for 18 h and tested for p-cresol production by gas-liquid chromatography. All 49 stock and reference strains of C. difficile and 19 organisms confirmed as C. difficile produced p-cresol. p-Cresol was not produced by 53 negative control or 161 test organi...

  12. Ultrasonic disruption of fungal mycelia for efficient recovery of polysaccharide-protein complexes from viscous fermentation broth of a medicinal fungus.

    Science.gov (United States)

    Cheung, Yi-Ching; Liu, Xing-Xun; Wang, Wing-Qiang; Wu, Jian-Yong

    2015-01-01

    High-intensity ultrasound (US) was applied to facilitate the extraction of intracellular and extracellular polysaccharide-protein complexes (PSPs) from the viscous mycelial fermentation broth of a medicinal fungus Cordyceps sinensis Cs-HK1. The US treatment caused the disruption of fungal mycelia, a dramatic reduction of the apparent broth viscosity, and the release of intracellular products into the liquid medium. The degree of mycelium disruption and the rate of intracellular product release were dependent on US power intensity, treatment period and biomass concentration of broth. The extraction or release kinetics of total water-soluble products and PSPs (yield Y versus time t) under the effect of US was fitted closely to the Elovich model Y=Yo+Y1 lnt and parabolic model Y=Yo+Y1t(½), respectively. Another interesting effect of the US treatment was a notable increase in the antioxidant cytoprotective activity of PSP against H2O2 induced cell death. PMID:24889549

  13. Evaluation of different buffered peptone water (BPW) based enrichment broths for detection of Gram-negative foodborne pathogens from various food matrices.

    Science.gov (United States)

    Margot, H; Zwietering, M H; Joosten, H; O'Mahony, Emer; Stephan, R

    2015-12-01

    This study evaluated the effects of changing the composition of the pre-enrichment medium buffered peptone water (BPW) on the growth of stressed and unstressed Gram-negative foodborne pathogens in a one-broth enrichment strategy. BPW supplemented with an available iron source and sodium pyruvate, along with low levels of 8-hydroxyquinoline and sodium deoxycholate (BPW-S) improved the recovery of desiccated Cronobacter spp. from powdered infant formula. Growth of Salmonella and STEC was comparable in all BPW variants tested for different food matrices. In products with high levels of Gram-negative background flora (e.g. sprouts), the target organisms could not be reliably detected by PCR in any of the BPW variants tested unless the initial level exceeded 10(3) cfu/10 g of sprouts. Based on these results we suggest BPW-S for a one-broth enrichment strategy of stressed Gram-negative foodborne pathogens from dry products. However, a one-broth enrichment strategy based on BPW variants tested in this evaluation is not recommended for produce with a high level of Gram-negative background flora due to very high detection limits. PMID:26267889

  14. Use of microdilution panels with and without beta-lactamase inhibitors as a phenotypic test for beta-lactamase production among Escherichia coli, Klebsiella spp., Enterobacter spp., Citrobacter freundii, and Serratia marcescens.

    Science.gov (United States)

    Thomson, K S; Sanders, C C; Moland, E S

    1999-06-01

    Over the past decade, a number of new beta-lactamases have appeared in clinical isolates of Enterobacteriaceae that, unlike their predecessors, do not confer beta-lactam resistance that is readily detected in routine antibiotic susceptibility tests. Because optimal methodologies are needed to detect these important new beta-lactamases, a study was designed to evaluate the ability of a panel of various beta-lactam antibiotics tested alone and in combination with beta-lactamase inhibitors to discriminate between the production of extended-spectrum beta-lactamases, AmpC beta-lactamases, high levels of K1 beta-lactamase, and other beta-lactamases in 141 isolates of Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, and Serratia marcescens possessing well-characterized beta-lactamases. The microdilution panels studied contained aztreonam, cefpodoxime, ceftazidime, cefotaxime, and ceftriaxone, with and without 1, 2, and 4 microg of clavulanate per ml or 8 microg of sulbactam per ml and cefoxitin and cefotetan with and without 8 microg of sulbactam per ml. The results indicated that a minimum panel of five tests would provide maximum separation of extended-spectrum beta-lactamase high AmpC, high K1, and other beta-lactamase production in Enterobacteriaceae. These included cefpodoxime, cefpodoxime plus 4 microg of clavulanate per ml, ceftazidime, ceftriaxone, and ceftriaxone plus 8 microg of sulbactam per ml. Ceftriaxone plus 2 microg of clavulanate per ml could be substituted for cefpodoxime plus 4 microg of clavulanate per ml without altering the accuracy of the tests. This study indicated that tests with key beta-lactam drugs, alone and in combination with beta-lactamase inhibitors, could provide a convenient approach to the detection of a variety of beta-lactamases in members of the family Enterobacteriaceae. PMID:10348759

  15. Comparative analysis of selected methods for the assessment of antimicrobial and membrane-permeabilizing activity: a case study for lactoferricin derived peptides

    Directory of Open Access Journals (Sweden)

    Lohner Karl

    2008-11-01

    Full Text Available Abstract Background Growing concerns about bacterial resistance to antibiotics have prompted the development of alternative therapies like those based on cationic antimicrobial peptides (APs. These compounds not only are bactericidal by themselves but also enhance the activity of antibiotics. Studies focused on the systematic characterization of APs are hampered by the lack of standard guidelines for testing these compounds. We investigated whether the information provided by methods commonly used for the biological characterization of APs is comparable, as it is often assumed. For this purpose, we determined the bacteriostatic, bactericidal, and permeability-increasing activity of synthetic peptides (n = 57; 9–13 amino acid residues in length analogous to the lipopolysaccharide-binding region of human lactoferricin by a number of the most frequently used methods and carried out a comparative analysis. Results While the minimum inhibitory concentration determined by an automated turbidimetry-based system (Bioscreen or by conventional broth microdilution methods did not differ significantly, bactericidal activity measured under static conditions in a low-ionic strength solvent resulted in a vast overestimation of antimicrobial activity. Under these conditions the degree of antagonism between the peptides and the divalent cations differed greatly depending on the bacterial strain tested. In contrast, the bioactivity of peptides was not affected by the type of plasticware (polypropylene vs. polystyrene. Susceptibility testing of APs using cation adjusted Mueller-Hinton was the most stringent screening method, although it may overlook potentially interesting peptides. Permeability assays based on sensitization to hydrophobic antibiotics provided overall information analogous – though not quantitatively comparable- to that of tests based on the uptake of hydrophobic fluorescent probes. Conclusion We demonstrate that subtle changes in methods for

  16. Identification of critical cell concentrations with in-situ and ex-situ characterization of physico-chemical properties of broth during oxidative axenic cultures of Yarrowia lipolytica in fed-batch mode

    OpenAIRE

    Kraiem, Hazar; Manon, Yannick; Anne-Archard, Dominique; Fillaudeau, Luc

    2012-01-01

    Identification of critical cell concentrations with in-situ and ex-situ characterization of physico-chemical properties of broth during oxidative axenic cultures of Yarrowia lipolytica in fed-batch mode

  17. Determination of Poly(malic acid) in Fermentation Broth by RP-HPLC%反向高效液相色谱测定发酵液中的聚苹果酸

    Institute of Scientific and Technical Information of China (English)

    乔长晟; 姜少丽; 马正旺; 李政; 楼鹏

    2012-01-01

    For the detection of poly (malic acid) fermentation broth, a reversed phase high performance liquid chromatography method was developed The samples was analyzed on a column of AlltimaTMC18 (4.6 mm× 150 mm, 4 um) with a mixture of acetonitrik and 0.025 mol/L KH2PO4 buffer (pH 2.5) (5:95, v:v) used as mobile phase at the flow rate of l.0mL/min. The detection wavelength was 210 nm; the column temperature was 25 ℃ and the volum of injection was 5 μL. Before sampling, the fermentation broth was centrifuged to remove fungis, added with organic solvent to remove polysaccharide and hydrolysed. L-malic acid were completely separated and determined in 3 minutes. The recovery and relative standard deviation (RSD, n=3) of L-malic acid by RP-HPLC method were 98.70%~100.65%, and 0.40%~1.11%, respectively. The recovery and relative standard deviation (RSD, n=3) of L-malic acid by L-malic acid kit were 96.70%~97.23% and 0.57%-1.19%, respectively. The method was proved to be a effective way to determine the poly (malic acid) in fermentation broth.%采用高效液相色谱柱AlltimaTM C18 (4.6 mm× 150 mm,4μm)上定量测定发酵生产的聚苹果酸,流动相为乙腈和0.025 mol/L磷酸二氢钾混合溶液(磷酸调pH 2.5)(V∶V=5∶ 95),流速1.0 mL/min,检测波长210 nm,柱温25℃,进样量5μL.发酵液经离心除菌体、纯沉除多糖、水解处理后进样,可以将其中的L苹果酸完全分离定量.采用液相方法的回收率为98.70%-100.65%,RSD为0.4-1.11%.采用试剂盒的方法的回收率为96.7~97.23%,RSD为0.57~1.19%.可见,液相的精确度和试剂盒相差不大.但液相的价格相对比试剂盒要便宜很多,说明该法是测定发酵液中聚苹果酸的一种很有效的方法.

  18. Growth and inactivation of Salmonella enterica and Listeria monocytogenes in broth and validation in ground pork meat during simulated home storage abusive temperature and home pan-frying

    Directory of Open Access Journals (Sweden)

    Xiang eWang

    2015-10-01

    Full Text Available Ground pork meat with natural microbiota and inoculated with low initial densities (1-10 or 10-100 CFU/g of Salmonella enterica or Listeria monocytogenes was stored under abusive temperature at 10°C and thermally treated by a simulated home pan-frying procedure. The growth and inactivation characteristics were also evaluated in broth. In ground pork meat, the population of S. enterica increased by less than one log after 12-days of storage at 10°C, whereas L. monocytogenes increased by 2.3 to 2.8 log units. No unusual intrinsic heat resistance of the pathogens was noted when tested in broth at 60°C although shoulders were observed on the inactivation curves of L. monocytogenes. After growth of S. enterica and L. monocytogenes at 10°C for 5 days to levels of 1.95 log CFU/g and 3.10 log CFU/g, respectively, in ground pork meat, their inactivation in the burger subjected to a simulated home pan-frying was studied. After thermal treatment S. enterica was undetectable but L. monocytogenes was recovered in three out of six of the 25 g burger samples. Overall, the present study shows that data on growth and inactivation of broths are indicative but may underestimate as well as overestimate behavior of pathogens and thus need confirmation in food matrix conditions to assess food safety in reasonably foreseen abusive conditions of storage and usual home pan-frying of of meat burgers in Belgium.

  19. Acetic Acid Production by an Electrodialysis Fermentation Method with a Computerized Control System

    OpenAIRE

    Nomura, Yoshiyuki; Iwahara, Masayoshi; Hongo, Motoyoshi

    1988-01-01

    In acetic acid fermentation by Acetobacter aceti, the acetic acid produced inhibits the production of acetic acid by this microorganism. To alleviate this inhibitory effect, we developed an electrodialysis fermentation method such that acetic acid is continuously removed from the broth. The fermentation unit has a computerized system for the control of the pH and the concentration of ethanol in the fermentation broth. The electrodialysis fermentation system resulted in improved cell growth an...

  20. Effect of an exo-polysaccharide from the culture broth of Hericium erinaceus on enhancement of growth and differentiation of rat adrenal nerve cells

    OpenAIRE

    Park, Young Shik; Lee, Hyun Soo; Won, Moo Ho; Lee, Jin Ha; Lee, Shin Young; Lee, Hyeon Yong

    2002-01-01

    It was found that an exo-biopolymer (M.W. 1,000,000, molar ratio of 1.5:1.7:1.2:0.6:0.9, glucose:galactose:xylose:mannose:fructose, purity 99%) purified from the liquid culture broth of Hericium erinaceus mycelium enhanced the growth of rat adrenal nerve cells. The polymer also improved the extension of the neurites of PC12 cell. Its efficacy was found to be higher than those from known nerve growth factors such as Nerve Growth Factor (NGF) and Brain-Derived Nerve Factor (BDNF). The effect of...

  1. 发酵液作为EBPR碳源的动力学模拟%Kinetic Simulation of Enhanced Biological Phosphorus Removal with Fermentation Broth as Carbon Source

    Institute of Scientific and Technical Information of China (English)

    张超; 陈银广

    2013-01-01

    发酵液是一种优质的碳源,能够提高生物除磷系统(EBPR)的除磷效果.采用基于碳源代谢的修正ASM2模型,能够较好地模拟发酵液作为EBPR碳源的动力学变化规律.发酵液作为EBPR唯一碳源时,系统中的异养菌不仅不对聚磷菌(PAO)的生长构成竞争关系,反而促进PAO的生长.发酵液作为实际污水的补充碳源时,优化了污水中的碳源组成,创造了有利于聚磷菌生长的环境,使EBPR中聚磷菌达到微生物总量的40%以上,比实际污水作为碳源的EBPR中的PAO含量提高了3.3倍.%As a high-quality carbon source,fermentation broth could promote the phosphorus removal efficiency in enhanced biological phosphorus removal (EBPR).The transformation of substrates in EBPR fed with fermentation broth was well simulated using the modified activated sludge model No.2 (ASM2) based on the carbon source metabolism.When fermentation broth was used as the sole carbon source,it was found that heterotrophic bacteria acted as a promoter rather than a competitor to the phosphorus accumulating organisms (PAO).When fermentation broth was used as a supplementary carbon source of real municipal wastewater,the wastewater composition was optimized for PAO growth; and the PAO concentration,which was increased by 3.3 times compared to that in EBPR fed with solely real municipal wastewater,accounting for about 40% of the total biomass in the reactor.

  2. Detection of Burkholderia pseudomallei in Sputum using Selective Enrichment Broth and Ashdown’s Medium at Kampong Cham Provincial Hospital, Cambodia [v2; ref status: indexed, http://f1000r.es/5e9

    Directory of Open Access Journals (Sweden)

    Somary Nhem

    2015-05-01

    Full Text Available Melioidosis, infection caused by Burkholderia pseudomallei, is increasingly reported in Cambodia. We hypothesized that implementation of an enhanced sputum testing protocol in a provincial hospital diagnostic microbiology laboratory would increase detection of B. pseudomallei. We tested 241 sputum specimens that were deemed acceptable for culture, comparing culture in selective enrichment broth followed by sub-culture on Ashdown’s medium to standard culture methods. Two specimens (0.8% were positive for B. pseudomallei using the enhanced protocol whereas one specimen (0.4% was positive using standard methods. Given the low numbers of positive specimens, we could not conclusively determine the utility of the enhanced sputum testing protocol. However, the ramifications of identification of B. pseudomallei are substantial, and the benefit of the enhanced testing protocol may be more apparent in patients selected based on risk factors and clinical presentation. Promoting clinician awareness of the infection and encouraging utilization of diagnostic microbiology services are also likely to be important factors in facilitating identification of melioidosis.

  3. Combined antimicrobial susceptibility test against pan-drug-resistant Acinetobacter baumannii with E-test and microdilution checkerboard assay%应用E-test法、肉汤微量稀释棋盘法检测泛耐药鲍曼不动杆菌的试验

    Institute of Scientific and Technical Information of China (English)

    王铁山; 苏建荣

    2013-01-01

    Objective To evaluate the activity of antibiotics against pan-drug-resistant (PDR) Acinetobacter baumannii by combination antimicrobial susceptibility test in viro with epsilometric methods (Etest method) and microdilution checkerboard (CB method),and to detect a good correlation between timekill curve with the above mentioned two assays.Methods Thirty-one clinical isolates of PDR Acinetobacter baumannii were selected for mono and combination antimicrobial susceptibility test in vitro by E-test and CB method,then a comparison was conducted between the test results and the time-kill curve.Mono drugs involved tigecycline,colistin,imipenem and amikacin,and combinations involved two of drugs above,and three drugs involved imipenem/tigecycline,plus amikacin combination.Results Synergistic effect was detected in imipenem plus colistin and tigecycline plus imipenem combination.A high comparability was revealed between the E-test method with antimicrobial drugs added into the culture medium and the time-kill curves.Synergy in the combination of imipenem/tigecycline,plus amikacin was detected by the CB method and time-kill curves.Conclusion The results showed that the effect of specific combination of antibiotics against PDR Acinetobacter baumannii could be predicted by testing their synergistic effect with combination antimicrobial susceptibility test.%目的 比较E-test法和肉汤微量稀释棋盘法(CB法)检测体外联合药敏试验并与时间杀菌曲线对比的观察.方法 选择31株泛耐药(PDR)鲍曼不动杆菌临床株进行体外试验,分为替加环素、可立其丁、亚胺培南、阿米卡星单用和两药组合,亚胺培南+替加环素+阿米卡星三药组合也由CB法测试.这些测试的结果与时间杀菌曲线进行比较.结果 亚胺培南+可立其丁、替加环素+亚胺培南能检测到协同作用.培养基中加入抗菌药物的E-test法联合药敏试验与时间杀菌曲线更具有可比性.在三药组合中由CB法和

  4. Enhanced production of bioethanol from waste of beer fermentation broth at high temperature through consecutive batch strategy by simultaneous saccharification and fermentation.

    Science.gov (United States)

    Khattak, Waleed Ahmad; Khan, Taous; Ha, Jung Hwan; Ul-Islam, Mazhar; Kang, Min-Kyung; Park, Joong Kon

    2013-10-10

    Malt hydrolyzing enzymes and yeast glycolytic and fermentation enzymes in the waste from beer fermentation broth (WBFB) were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A new 'one-pot consecutive batch strategy' was developed for efficient bio-ethanol production by simultaneous saccharification and fermentation (SSF) using WBFB without additional enzymes, microbial cells, or carbohydrates. Bio-ethanol production was conducted in batches using WBFB supernatant in the first phase at 25-67°C and 50rpm, followed by the addition of 3% WBFB solid residue to the existing culture broth in the second phase at 67°C. The ethanol production increased from 50 to 102.5g/L when bare supernatant was used in the first phase, and then to 219g ethanol/L in the second phase. The amount of ethanol obtained using this strategy was almost equal to that obtained using the original WBFB containing 25% solid residue at 33°C, and more than double that obtained when bare supernatant was used. Microscopic and gel electrophoresis studies revealed yeast cell wall degradation and secretion of cellular material into the surrounding medium. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) supported the existence of enzymes in WBFB involved in bioethanol production at elevated temperatures. The results of this study will provide insight for the development of new strategies for biofuel production. PMID:24034431

  5. Quantitative Determination of Dihydroxyacetone and Glycerol in the Fermentation Broth by HPLC%HPLC检测发酵液中二羟基丙酮和甘油的含量

    Institute of Scientific and Technical Information of China (English)

    谢光蓉

    2013-01-01

    High performance liquid chromatography (HPLC) was applied to determinate dihydroxyacetone (DHA) and glycerol in the fermentation broth respectively. The mobile phase is composed of acetonitrile-water (85∶15, V∶V). The analytes were separated on a Hedera-NH2 column with at a flow rate of 1mL/min. DHA was detected by UV detector at 271nm and glycerol was detected by refractometer. The linearity range for the determination of DHA was 2.00-12.00mg/mL with a correlation coefficient (r) of 0.9996. The linearity range for the determination of glycerol was 0.1-1.0mg/mL with a correlation coefficient (r) of 0.9998. The methods were applicable for DHA and glycerol determination in the fermentation process without interference from other constitutes in the fermentation broth.%  应用高效液相色谱法检测发酵过程中发酵产物二羟基丙酮和底物甘油浓度。在流动相乙腈、纯净水的比例为85∶15(v∶v),流速为1mL/min,色谱柱为Hedera-NH2的条件下,用紫外检测器在271nm处检测DHA,用示差折光检测器检测甘油。高效液相色谱法检测DHA的线性范围为2.0-12.0mg/mL,相关系数(r)为0.9996,检测甘油的线性范围为0.1-1.0mg/mL,相关系数(r)为0.9998。该方法应用于二羟基丙酮发酵过程中底物和产物的检测,该方法不受培养基中不同碳源(葡萄糖、蔗糖、山梨醇、乳糖、甘油),不同氮源(蛋白胨、酵母膏、酵母粉、营养肉汤)以及菌体未完全利用的培养基成分、菌体生长过程中产生的其它代谢产物的影响,该方法具有较好的通用性。

  6. Study on L-Valine Efficient Extraction Technology from the Fermentation Broth%从发酵液中高效提取L-缬氨酸的工艺研究

    Institute of Scientific and Technical Information of China (English)

    李新涛; 徐庆阳; 冯宁; 谢希贤; 陈宁

    2011-01-01

    目的:利用有机膜过滤和离子交换法分离提取发酵液中的L-缬氨酸.方法:通过有机膜过滤,除去发酵液中的菌体及蛋白,滤液浓缩结晶获得L-缬氨酸产品,通过离子交换法从结晶母液中回收部分L-缬氨酸.结果:确定了有机微滤膜和超滤膜去除发酵液中菌体蛋白和色素的操作条件;确定了采用离子交换法提取L-缬氨酸的操作条件:选择732强酸性阳离子树脂,料液pH值为3.0左右,用0.4 mol/L的氨水以1.0 mL/min的速度洗脱,L-缬氨酸的收率为89.2%.结论:通过有机膜过滤和离子交换法分离提取发酵液中的L-缬氨酸,可以提高提取收率和产品质量.%Objective: Using organic membrane filtration and ion exchange to extract L-valine from the fermentation broth. Methods: The bacteria and proteins in the fermentation broth were removed through organic membrane filtration, the filtrate was concentrated to obtain L-valine crystal products, the part of L-valine was further extracted from the crystal liquor by ion exchange. Results: The optimum conditions of organism membrane filtration, decolour and abstracton of L-valine were determined. 732 resin was the optimal resin for L-valine abstraction among the different kinds of resins. The optimal conditions of adsorption were as follow: dosage liquor pH3.0, and the elution of L-valine was carried out at the rate of 1.0 mL/min using 0.4 mol/L ammonia water as eluant. The recovery of L-valine was up to 89.2%. Conclution: L-valine extraction yield and product quality was improved by the way of organic membrane filtration and ion exchange.

  7. Antifungal activity of the essential oils of wild-growing Mentha piperita L and Mentha spicata L from the Mariovo region, Republic of Macedonia

    OpenAIRE

    Kostik, Vesna; Bauer, Biljana

    2015-01-01

    The antifungal activity of the essential oils isolated by hydro-distillation from the leaves of wild growing Mentha piperita and Mentha spicata (Lamiaceae) at the region of Mariovo, Republic of Macedonia was tested by disc diffusion method and the micro-dilution broth method (MIC) against six plant pathogenic fungi: Alternaria alternate Alternaria solani, Aspergillus flavus, Aspergillus niger, Fusarium solani and Rhizopus solani. The results from the disc diffusion method followed by MIC indi...

  8. Evaluation of Methods To Increase the Sensitivity and Timeliness of Detection of Streptococcus agalactiae in Pregnant Women

    OpenAIRE

    Overman, Sue B.; Eley, Douglas D.; Jacobs, Barry E.; Ribes, Julie A.

    2002-01-01

    Direct culture of rectovaginal specimens on Granada agar was compared to culture on sheep blood agar plate (SBAP) and AccuProbe detection of group B streptococcus from overnight LIM broth enhancement cultures (LIM-SBAP). Both broth-enhanced methods demonstrated excellent sensitivity (97.5% for LIM-SBAP and 93.5% for AccuProbe), while Granada agar demonstrated a sensitivity of only 40.3%.

  9. Antimicrobial Activity of Untenospongin B, a Metabolite from the Marine Sponge Hippospongia communis collected from the Atlantic Coast of Morocco

    Directory of Open Access Journals (Sweden)

    Aziz Fassouane Fassouane

    2004-08-01

    Full Text Available Abstract: (--Untenospongin B isolated from the marine sponge Hippospongia communis has been tested for its antimicrobial activity against bacteria and human pathogenic fungi using agar disk method and was found to possess a broad and strong activity toward the test organisms. Its antifungal activity was further characterized by determination of the minimum inhibitory concentration (MIC against five fungal species using broth microdilution method.

  10. Rapid detection of polymyxin-resistant Enterobacteriaceae from blood cultures

    OpenAIRE

    Jayol, Aurélie; Dubois, Véronique; Poirel, Laurent; Nordmann, Patrice

    2016-01-01

    Enterobacterial strains resistant to polymyxins are increasingly reported worldwide. Conventional methods for detection of colistin-resistant isolates such as broth microdilution remain time-consuming (24 to 48 h), and methods such as disc diffusion and E-test are not reliable. Recently, the Rapid Polymyxin NP test was developed for a rapid identification of polymyxin-resistant Enterobacteriaceae. This test is based on the detection of glucose metabolism related to bacterial growth in th...

  11. Pimenta pseudocaryophyllus inhibits virulence factors and promotes metabolic changes in Candida yeast

    OpenAIRE

    2014-01-01

    Introduction This is the first study to examine the in vitro susceptibility and the expression of virulence factors in Candida species in the presence of Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae), a Brazilian plant known as paucravo. Additionally, the mechanisms of action of the crude ethanol extract and the ethyl acetate and aqueous fractions of this plant were investigated. Methods The in vitro susceptibility of Candida was tested using the broth microdilution method, wh...

  12. Manuka honey treatment of biofilms of Pseudomonas aeruginosa results in the emergence of isolates with increased honey resistance

    OpenAIRE

    Camplin, Aimee L; Maddocks, Sarah E.

    2014-01-01

    Background Medical grade manuka honeys are well known to be efficacious against Pseudomonas aeruginosa being bactericidal and inhibiting the development of biofilms; moreover manuka honey effectively kills P. aeruginosa embedded within an established biofilm. Sustained honey resistance has not been previously documented for planktonic or biofilm P. aeruginosa. Methods Minimum inhibitory concentrations for manuka honey and antibiotics were determined using broth micro-dilution methods. Minimum...

  13. Microwave-assisted multi-component synthesis of 3’-indolyl substituted pyrano[2,3-c]pyrazoles and their antimicrobial activity

    Directory of Open Access Journals (Sweden)

    Kathrotiya Harshad G.

    2012-01-01

    Full Text Available A series of pyrano[2,3-c]pyrazole derivatives of indole has been synthesized by multi-component reaction using conventional and microwave irradiation approach. Particularly valuable features of this method include high yield, broad substrate scope, shorter reaction time and straightforward procedure. Antimicrobial screening against eight human pathogens, namely B. subtilis, C. tetani, S. pneumoniae, S. typhi, V. cholerae, E. coli, A. fumigatus and C. albicans by employing broth microdilution MIC method as recommended by NCCLS.

  14. Proanthocyanidins polymeric tannin from Stryphnodendron adstringens are active against Candida albicans biofilms

    OpenAIRE

    Luiz, Raul Leal Faria; Vila, Taissa Vieira Machado; de Mello, João Carlos Palazzo; Nakamura, Celso Vataru; Rozental, Sonia; Ishida, Kelly

    2015-01-01

    Background Biofilm formation is important in Candida albicans pathogenesis and constitutes a mechanism of antifungal resistance. Thus, we evaluated the effect of proanthocyanidins polymer-rich fractions from Stryphnodendron adstringens (fraction F2 and subfraction F2.4) against C. albicans biofilms. Methods Firstly, the antifungal activity of F2 and F2.4 against planktonic cells of Candida albicans (ATCC 10231) was determined using broth microdilution method. Anti-biofilm effect of F2 and F2....

  15. Dominance of CTX-M-Type Extended-Spectrum β-Lactamase (ESBL)-Producing Escherichia coli Isolated from Patients with Community-Onset and Hospital-Onset Infection in China

    OpenAIRE

    Xia, Shu; Fan, Xin; Huang, Zengguang; XIA, LIANG; Xiao, Meng; Chen, Rongchang; Xu, Yingchun; Zhuo, Chao

    2014-01-01

    Objective To investigate CTX-M genotypes among extended-spectrum β-lactamase-producing Escherichia coli (ESBL-EC) isolated from patients with community-onset and hospital-onset infections in China, their clonality and the distribution of CTX-M variants in different specimens of community-onset and hospital-onset infections. Methods ESBL-EC isolates were collected from general hospitals from 2011 to 2012 in China. Broth microdilution method antimicrobial susceptibility testing of 16 antibiotic...

  16. Synergistic effect of xylitol and ursolic acid combination on oral biofilms

    OpenAIRE

    Zou, Yunyun; Lee, Yoon; Huh, Jinyoung; Park, Jeong-Won

    2014-01-01

    Objectives This study was designed to evaluate the synergistic antibacterial effect of xylitol and ursolic acid (UA) against oral biofilms in vitro. Materials and Methods S. mutans UA 159 (wild type), S. mutans KCOM 1207, KCOM 1128 and S. sobrinus ATCC 33478 were used. The susceptibility of S. mutans to UA and xylitol was evaluated using a broth microdilution method. Based on the results, combined susceptibility was evaluated using optimal inhibitory combinations (OIC), optimal bactericidal c...

  17. Antifungal activity of triterpenoid isolated from Azima tetracantha leaves.

    OpenAIRE

    Duraipandiyan, V; M Gnanasekar; S Ignacimuthu

    2010-01-01

    The present study was designed to evaluate the antifungal activity of Azima tetracantha extracts and isolated compound (friedelin) against fungi. Antifungal activity was carried out using broth microdilution method and fractions were collected using (silica gel) column chromatography. The antifungal activity of Azima tetracantha crude extracts and isolated compound (friedelin) were evaluated using the micro dilution method. Hexane extract showed some antifungal activity. The compound also exh...

  18. Adequacy of Petrifilm™ Aerobic Count plates supplemented with de Man, Rogosa & Sharpe broth and chlorophenol red for enumeration of lactic acid bacteria in salami.

    Science.gov (United States)

    de Castilho, Natália Parma Augusto; Okamura, Vivian Tiemi; Camargo, Anderson Carlos; Pieri, Fábio Alessandro; Nero, Luís Augusto

    2015-12-01

    The present study aimed to assess the performance of alternative protocols to enumerate lactic acid bacteria (LAB) in salami. Fourteen cultures and two mixed starter cultures were plated using six protocols: 1) Petrifilm™ Aerobic Count (AC) with MRS broth and chlorophenol red (CR), incubated under aerobiosis or 2) under anaerobiosis, 3) MRS agar with CR, 4) MRS agar with bromocresol purple, 5) MRS agar at pH5.7, and 6) All Purpose Tween agar. Samples of salami were obtained and the LAB microbiota was enumerated by plating according protocols 1, 2, 3 and 5. Regression analysis showed a significant correlation between the tested protocols, based on culture counts (p0.05). Colonies were confirmed as LAB, indicating proper selectivity of the protocols. The results showed the adequacy of Petrifilm™ AC supplemented with CR for the enumeration of LAB in salami. PMID:26291606

  19. Listeria monocytogenes efficiently invades caco-2 cells after low-temperature storage in broth and on deli meat

    DEFF Research Database (Denmark)

    Larsen, Marianne Halberg; Koch, Anette Granly; Ingmer, Hanne

    2010-01-01

    The objective of this study was to investigate how various growth conditions influence the virulence of Listeria monocytogenes monitored by its ability to invade the epithelial cell lines Caco-2 and INT-407. The growth conditions examined were modified atmosphere-packaged deli meat and brain heart...... infusion broth (BHI) with and without salt. Five strains of L. monocytogenes were selected to investigate their invasiveness and all strains invaded Caco-2 cells at higher levels than INT-407 cells. Further, the clinical strains (3443 and 3734) were more invasive (p < 0.05) than the strains isolated from...... meat and food-processing environments (3008, 3126, and 4140) after grown in BHI at 30 degrees C. This attenuation could not be ascribed to a defective Internalin A as all strains encoded an intact inlA gene. To determine the influence of food products on virulence, the ability of L. monocytogenes to...

  20. Discovery, production and purification of the Na+, K+ activated ATPase inhibitor, L-681,110 from the fermentation broth of Streptomyces sp. MA-5038.

    Science.gov (United States)

    Huang, L; Albers-Schonberg, G; Monaghan, R L; Jakubas, K; Pong, S S; Hensens, O D; Burg, R W; Ostlind, D A; Conroy, J; Stapley, E O

    1984-09-01

    The maximum yield for the production of L-681,110 by Streptomyces sp. MA-5038 (ATCC 31587) was observed after 5 days' incubation at 28 degrees C and pH about 8.3. L-681,110 was isolated from the fermentation broth by acetone extraction of the mycelia, absorption to Amberlite XAD-2 resin and two separations by thin-layer chromatography. The structure of L-681,110 was found to consist of a sixteen-membered lactone with a new type of substitution. The inhibition of ATPase, activity against Caenorhabditis elegans and stimulation of gamma-aminobutyric acid release indicate that L-681,110 possesses some characteristics of both oligomycin and avermectin. L-681,110 was also active against tapeworm and ticks in an in vivo assay. PMID:6094416

  1. A novel laccase with inhibitory activity towards HIV-I reverse transcriptase and antiproliferative effects on tumor cells from the fermentation broth of mushroom Pleurotus cornucopiae.

    Science.gov (United States)

    Wu, Xiangli; Huang, Chenyang; Chen, Qiang; Wang, Hexiang; Zhang, Jinxia

    2014-04-01

    A novel laccase with a molecular mass of 67 kDa was isolated from the fermentation broth of Pleurotus cornucopiae through ion exchange chromatography and gel filtration. The optimal pH and temperature for the laccase was pH 4.2 and 30°C, respectively. The laccase activity was remarkably inhibited by Fe(3+) and Hg(2+) , while it was stimulated by Cu(2+) and Pb(2+) . It inhibited proliferation of the hepatoma cells HepG2 and the breast cancer cells MCF-7, and the activity of HIV-I reverse transcriptase with IC50 values of 3.9, 7.6 and 3.7 μM, respectively. PMID:24136666

  2. Membrane Separation Process of Oxytetracycline Fermentation Broth with Magnetization%磁场强化膜分离土霉素发酵液工艺研究

    Institute of Scientific and Technical Information of China (English)

    王亚卿

    2012-01-01

    The oxytetracycline filtered broth by plate and framefilter was purified and concentrated with membrane separation techniques in magnetic field.Experiment results indicate that ultrafiltration membrane PES20 and nanofiltration membrane NF270 separation properties is better effect in 0.4 T,and the production quality of oxytetracycline increased compared with no magnetization treatment process or the original technology.%引入磁场强化膜分离技术对板框过滤后的土霉素发酵液进行提纯和浓缩,研究结果表明,经0.4T磁段磁场强化后,PES20超滤膜及NF270纳滤分离性能均表现出较佳的正效应,土霉素产品质量较无磁场作用及原工艺有所提高。

  3. Development and Optimization of a Novel Immunomagnetic Separation- Bacteriophage Assay for Detection of Salmonella enterica Serovar Enteritidis in Broth

    OpenAIRE

    Favrin, Stacy J.; Jassim, Sabah A.; Griffiths, Mansel W

    2001-01-01

    Salmonella is the second-leading cause of food-borne illness in most developed countries, causing diarrhea, cramps, vomiting, and often fever. Many rapid methods are available for detection of Salmonella in foods, but these methods are often insensitive or expensive or require a high degree of technical ability to perform. In this paper we describe development and characterization of a novel assay that utilizes the normal infection cycle of bacteriophage SJ2 for detection of Salmonella enteri...

  4. Análise sensorial de caldos e canjas elaborados com farinha de carcaças de peixe defumadas: aplicação na merenda escolar Sensorial analysis of soups and broths made by smoked fish carcasses meal: its utilization to supplement school meals

    Directory of Open Access Journals (Sweden)

    Leandro Cesar de Godoy

    2010-05-01

    Full Text Available O trabalho avaliou a aceitação de caldos e canjas elaborados com farinhas aromatizadas desenvolvidas a partir de carcaças de peixes defumadas. As carcaças de tilápia do Nilo, carpa e pacu foram lavadas, identificadas, pesadas, imersas em salmoura com ervas aromáticas e defumadas. O produto defumado foi submetido à moagem para obtenção das farinhas, a partir das quais foram elaborados o caldo e a canja. Porções das farinhas, dos caldos e das canjas foram avaliadas por um painel de 40 provadores utilizando-se um método de estímulo simples, sendo avaliados os atributos: aroma, sabor, cor, textura, aparência e aceitação geral. Não houve diferença significativa (P > 0,05 na aceitação geral dos produtos. Os caldos elaborados com estas farinhas tiveram uma excelente aceitação, não diferindo significativamente entre si no que se refere aos atributos avaliados. A canja elaborada a partir da farinha de carcaça de pacu apresentou as menores notas quando comparada às demais canjas. Portanto, as farinhas aromatizadas podem ser empregadas no enriquecimento de produtos para o consumo humano. Esses produtos podem ser aplicados na merenda escolar, melhorando a qualidade nutricional das refeições. Além disso, tal uso daria um destino nobre aos resíduos que podem causar sérios impactos se descartados no meio ambiente.This work evaluated the acceptance of soups and broths prepared with aromatized meals made from smoked fish carcasses. The species of fish used for smoking were Nile Tilapia , carp, and pacu. The carcasses were washed, labeled, weighted, immersed in a solution of brine with aromatic herbs, and smoked. The smoked product was crushed to obtain the meal, which was used to cook the soups and the broths . Portions of meals, broths and soups were sampled by 40 tasters based on a simple stimulus method, which evaluated characteristics such as aroma, flavor, color, texture, aspect, and general acceptance. Considering the three

  5. ANTIFUNGAL ACTIVITY OF NEEM (Azadirachta indica: MELIACEAE) EXTRACTS AGAINST DERMATOPHYTES

    OpenAIRE

    Ospina Salazar, Daniel Iván; Hoyos Sánchez, Rodrigo Alberto; Fernando OROZCO SÁNCHEZ; Myrtha ARANGO ARTEAGA; Luisa Fernanda GÓMEZ LONDOÑO

    2015-01-01

    In order to assess the antifungal activity of methanolic extracts from neem tree (Azadirachta indica A. Juss.), several bioassays were conducted following M38-A2 broth microdilution method on 14 isolates of the dermatophytes Trichophyton mentagrophytes, Trichophyton rubrum, Microsporum canis and Epidermophyton floccosum. Neem extracts were obtained through methanol-hexane partitioning of mature green leaves and seed oil. Furthermore, high performance liquid chromatography (HPLC) analyses were...

  6. In vitro sensitivity of poultry Brachyspira isolates to essential oil components and in vivo reduction of Brachyspira intermedia in rearing pullets with cinnamaldehyde feed supplementation

    OpenAIRE

    Verlinden, Marc; Pasmans, Frank; Mahu, Maxime; Vande Maele, Lien; DE PAUW, Nele; Yang, Zhen; Haesebrouck, Freddy; Martel, An

    2013-01-01

    Cecal enteritis due to Brachyspira infections tends to be chronic in laying hens. Limited availability of antimicrobial drugs for use in laying hens emphasizes the need for alternative control measures. A broth microdilution method was used to determine the antimicrobial susceptibility of 20 Brachyspira intermedia field isolates from laying hen flocks to components of essential oils (EO). Minimal inhibitory concentration (MIC) distributions, obtained for eight EO components, were all monomoda...

  7. Methodological comparisons for antimicrobial resistance surveillance in feedlot cattle

    OpenAIRE

    Katharine M Benedict; Gow, Sheryl P.; Checkley, Sylvia; Booker, Calvin W.; McAllister, Tim A; Morley, Paul S.

    2013-01-01

    Background The purpose of this study was to objectively compare methodological approaches that might be utilized in designing an antimicrobial resistance (AMR) surveillance program in beef feedlot cattle. Specifically, four separate comparisons were made to investigate their potential impact on estimates for prevalence of AMR. These included investigating potential differences between 2 different susceptibility testing methods (broth microdilution and disc diffusion), between 2 different targ...

  8. Characterization of Anticancer, Antimicrobial, Antioxidant Properties and Chemical Compositions of Peperomia Pellucida Leaf Extract

    OpenAIRE

    Desy Fitrya Syamsumir; Julius Yong Fu Siong; Wendy Wee; Lee Seong Wei

    2011-01-01

    Peperomia pellucida leaf extract was characterized for its anticancer, antimicrobial, antioxidant activities, and chemical compositions. Anticancer activity of P. pellucida leaf extract was determined through Colorimetric MTT (tetrazolium) assay against human breast adenocarcinoma (MCF-7) cell line and the antimicrobial property of the plant extract was revealed by using two-fold broth micro-dilution method against 10 bacterial isolates. Antioxidant activity of the plant extract was then char...

  9. In vitro activity of antimicrobial agents against oxacillin resistant staphylococci with special reference to Staphylococcus haemolyticus

    OpenAIRE

    Chaudhury A; Kumar A

    2007-01-01

    One hundred and sixty seven isolates of staphylococci isolated from the inpatients of a tertiary care referral hospital in South India were speciated and activity of oxacillin, glycopeptides, linezolid and quinupristin/dalfopristin against these isolates was tested by broth microdilution method. Of the 114 coagulase negative staphylococci (CoNS), 49.1 % were S. haemolyticus, isolated predominantly from urine (64.6%), while the rest belonged to 11 other species. More than half ...

  10. In vitro activity of biapenem (L-627), a new carbapenem, against anaerobes.

    OpenAIRE

    Aldridge, K E; MORICE,N; Schiro, D D

    1994-01-01

    We tested 441 clinical strains of anaerobes by using a broth microdilution method to determine the in vitro activity of biapenem for comparison with those of other agents. Biapenem had activity comparable to those of imipenem and meropenem against all groups of anaerobes with MICs for 90% of the strains tested of 0.06 to 2 micrograms/ml. Against the Bacteroides fragilis group, biapenem was more active than ampicillin-sulbactam, ticarcillin-clavulanate, piperacillin, cefoxitin, cefotaxime, and...

  11. Synthesis and In Vitro Evaluation of New Thiosemicarbazone Derivatives as Potential Antimicrobial Agents

    OpenAIRE

    Zafer Asım Kaplancıklı; Mehlika Dilek Altıntop; Belgin Sever; Zerrin Cantürk; Ahmet Özdemir

    2016-01-01

    In an effort to develop potent antimicrobial agents, new thiosemicarbazone derivatives were synthesized via the reaction of 4-[4-(trifluoromethyl)phenyl]thiosemicarbazide with aromatic aldehydes. The compounds were evaluated for their inhibitory effects on pathogenic bacteria and yeasts using the CLSI broth microdilution method. Microplate Alamar Blue Assay was also carried out to determine the antimycobacterial activities of the compounds against Mycobacterium tuberculosis H37Rv. Among these...

  12. The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

    OpenAIRE

    Marina Azevedo Souza; Susana Johann; Luciana Alves Rodrigues dos Santos Lima; Fernanda Fraga Campos; Isolda Castro Mendes; Heloisa Beraldo; Elaine Maria de Souza-Fagundes; Patricia Silva Cisalpino; Carlos Augusto Rosa; Tania Maria de Almeida Alves; Nivea Pereira de Sa; Carlos Leomar Zani

    2013-01-01

    Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs) of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazone an...

  13. The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

    OpenAIRE

    2013-01-01

    Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs) of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazo...

  14. Antimicrobial and Antioxidant Activities of Two Endemic Plants from Aksaray in Turkey

    OpenAIRE

    Ozusaglam, Meltem Asan; Darilmaz, Derya Onal; Erzengin, Mahmut; Teksen, Mehtap; Erkul, Seher Karaman

    2013-01-01

    This study was designed to examine the in vitro antimicrobial and antioxidant activities of the methanol, ethanol, water, n-hexane and dicholoromethane extracts of two Allium species (Allium scabriflorum and Allium tchihatschewii) which are endemic for the flora of Turkey. The antimicrobial efficiency of the plant was evaluated according to disc diffusion and microdilution broth methods. The antimicrobial test results showed that the extracts of A. scabriflorum and A. tchihatschewii showed va...

  15. Efficacy of Cuminum cyminum essential oil on FUM1 gene expression of fumonisin-producing Fusarium verticillioides strains

    OpenAIRE

    Ali Reza Khosravi; Hojjatollah shokri; Ali Reza Mokhtari

    2014-01-01

    Objectives: The purpose of this study was to evaluate the effect of Cuminum cyminum (C. cyminum) essential oil on the growth and FUM1 gene expression of fumonisin-producing Fusarium verticillioides (F. verticillioides) strains isolated from maize. Materials and Methods: All fungal strains were cultured on potato dextrose agar (PDA) slopes at 30°C for 7 days. The antifungal activity was evaluated by broth microdilution assay. One set of primers was F. verticillioides species specific, which se...

  16. Antibacterial and anti-inflammatory activities of an extract, fractions, and compounds isolated from Gochnatia pulchra aerial parts

    OpenAIRE

    Lucarini, R.; Tozatti, M.G.; Silva, M.L.A.; V.M.M. Gimenez; P.M. Pauletti; M. Groppo; Turatti, I.C.C.; Cunha, W. R.; C.H.G. Martins

    2015-01-01

    This paper reports on the in vitro antibacterial and in vivo anti-inflammatory properties of a hydroethanolic extract of the aerial parts of Gochnatia pulchra (HEGP). It also describes the antibacterial activity of HEGP fractions and of the isolated compounds genkwanin, scutellarin, apigenin, and 3,5-O-dicaffeoylquinic acid, as evaluated by a broth microdilution method. While HEGP and its fractions did not provide promising results, the isolated compounds exhibited pronounced antibacterial ac...

  17. Chemical composition and antibacterial activity of Cochlospermum planchoni Hook.f. ex Planch essential oil from Burkina Faso.

    Science.gov (United States)

    Ouattara, Lassina; Koudou, Jean; Obame, Louis C E; Karou, Damintoti S; Traore, Alfred; Bessière, Jean Marie

    2007-11-15

    The water distilled oil obtained from rhizomes of Cochlospermum planchonii Hook.f.ex Planch (Apocynaceae) from Burkina Faso was examined by GC and GC/MS. Cochlospermum planchonii oil presents a particular chemical composition with a high rate of oxygenated components with predominance of ketones and esters (86.4%). The essential oil was tested against twelve strains of bacteria using a broth microdilution method. The results suggest that Cochlospermum planchonii essential oil has significant bactericidal activity. PMID:19090305

  18. Chemical Composition and Antimicrobial Activity of Leaf, Ripe and Unripe Peel of Bitter Orange (Citrus aurantium) Essential Oils

    OpenAIRE

    Fatemeh Azhdarzadeh; Mohammad Hojjati

    2016-01-01

    Background and Objectives: The chemical composition and antimicrobial activity of bitter orange (Citrus aurantium) leaf, ripe and unripe peel essential oils, cultivated in southwest of Iran, were investigated. Materials and Methods: The analysis of chemical composition of hydro-distilled essential oils was carried out by GC-MS. The disc diffusion and broth micro-dilution were used to assay the antimicrobial effect of achieved essential oils. Results: According to the GC-MS analysis, 3...

  19. Essential oil of Melaleuca alternifolia for the treatment of oral candidiasis induced in an immunosuppressed mouse model

    OpenAIRE

    de Campos Rasteiro, Vanessa Maria; da Costa, Anna Carolina Borges Pereira; Araújo, Cássia Fernandes; de Barros, Patrícia Pimentel; Rossoni, Rodnei Dennis; Anbinder, Ana Lia; Jorge, Antonio Olavo Cardoso; Junqueira, Juliana Campos

    2014-01-01

    Background The search for alternative therapies for oral candidiasis is a necessity and the use of medicinal plants seems to be one of the promising solutions. The objective of this study was to evaluate the in vitro and in vivo effects of the essential oil of Melaleuca alternifolia on Candida albicans. Methods The minimum inhibitory concentration (MIC) and minimum biofilm eradication concentration (MBEC) of M. alternifolia were determined by the broth microdilution assay. For the in vivo stu...

  20. In vitro antifungal activity of hydroxychavicol isolated from Piper betle L

    OpenAIRE

    Afrin Farhat; Dutt Prabhu; Satti Naresh K; Gupta Bishan D; Suri Krishan A; Khan Farrah G; Ali Intzar; Qazi Ghulam N; Khan Inshad A

    2010-01-01

    Abstract Background Hydroxychavicol, isolated from the chloroform extraction of the aqueous leaf extract of Piper betle L., (Piperaceae) was investigated for its antifungal activity against 124 strains of selected fungi. The leaves of this plant have been long in use tropical countries for the preparation of traditional herbal remedies. Methods The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of hydroxychavicol were determined by using broth microdilution ...

  1. Isolierungsraten, Resistenzverhalten und genetische Diversität von Campylobacter aus Tierbeständen und Umwelthabitaten

    OpenAIRE

    Löwenstein, Anna Elisabeth

    2016-01-01

    The aim of the study was to identify potential sources of Campylobacter spp. and their prevalences in Greater Berlin area from November 2011 to June 2013. Samples collected from pet animals, farm animals, wild animals and the environment were investigated by two isolation methods – ISO (according to ISO 10272-1:2006) and the Cape Town protocol. Also the presence of antimicrobial resistance in selected C. jejuni and C. coli isolates was determined by Broth Microdilution assays and isolates ...

  2. Antimicrobial, antioxidant, and cytotoxic activities of Bixa orellana Linn

    OpenAIRE

    Ul Islam, S.M. Bakhtiar; Hossain, Khalid; Gomes, Isidore; Gomes, Donald J.; Rahman, Sabita Rezwana; Mohammad S. Rahman; RASHID, MOHAMMAD A.

    2011-01-01

    Bixa orellana Linn., commonly known as "lipstick plant", is used in folk medicines to treat infections of microbial origin as well as coloring agents in food stuffs in the LDCs like Bangladesh. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the warm water extract of leaves of B. orellana were evaluated against 25 multidrug resistant (MDR) clinical isolates and 6 food-borne pathogens using the micro-dilution broth method modified to comply with the N...

  3. In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae) against oral pathogens=Estudo in vitro do efeito antimicrobiano dos extratos hidroalcólicos de Mentha arvensis L. (Lamiaceae) contra patógenos orais

    OpenAIRE

    Rogério Antonio Freitag; Gladis Aver Ribeiro; Patrícia da Silva Nascente; Rosana Serpa; Rafael Guerra Lund; Francisco Augusto Burkert Del Pino

    2012-01-01

    In vitro tests could be a valuable tool for the evaluation of medicinal plants’ antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods) were used to evaluate antimicrobial activity. Although hy...

  4. In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae) against oral pathogens - doi: 10.4025/actascibiolsci.v34i4.8959

    OpenAIRE

    Francisco Augusto Burkert Del Pino; Gladis Aver Ribeiro; Rogério Antonio Freitag; Patrícia da Silva Nascente; Rosana Serpa; Rafael Guerra Lund

    2012-01-01

    In vitro tests could be a valuable tool for the evaluation of medicinal plants’ antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods) were used to evaluate antimicrobial activity. Although hy...

  5. Modified ecometric technique (four-quadrant sequential streak) to evaluate Campylobacter enrichment broth proficiency in suppressing background microflora

    Science.gov (United States)

    Ecometric technique is a semi-quantitative scoring method used for quality control of culture media in microbiological laboratories. The technique involves inoculation with defined populations of specific culture onto solid media via a standardized chronological streaking technique, leading to ever-...

  6. Influence of aerobic and anaerobic conditions on survival of Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium in Luria-Bertani broth, farm-yard manure and slurry

    NARCIS (Netherlands)

    Semenov, A.V.; Overbeek, van L.S.; Termorshuizen, A.J.; Bruggen, van A.H.C.

    2011-01-01

    The influence of aerobic and anaerobic conditions on the survival of the enteropathogens Escherichia coli O157:H7 and Salmonella serovar Typhimurium was investigated in microcosms with broth, cattle manure or slurry. These substrates were inoculated with a green fluorescent protein transformed strai

  7. Influence of aerobic and anaerobic conditions on survival of Escherichia coli O157 : H7 and Salmonella enterica serovar Typhimurium in Luria-Bertani broth, farm-yard manure and slurry

    NARCIS (Netherlands)

    Semenov, Alexander V.; van Overbeek, Leo; Termorshuizen, Aad J.; van Bruggen, Ariena H. C.

    2011-01-01

    The influence of aerobic and anaerobic conditions on the survival of the enteropathogens Escherichia coli O157:H7 and Salmonella serovar Typhimurium was investigated in microcosms with broth, cattle manure or slurry. These substrates were inoculated with a green fluorescent protein transformed strai

  8. Methicillin-Resistant Staphylococcus aureus in Spain: Molecular Epidemiology and Utility of Different Typing Methods ▿ †

    OpenAIRE

    Vindel, Ana; Cuevas, Oscar; Cercenado, Emilia; Marcos, Carmen; Bautista, Verónica; Castellares, Carol; Trincado, Pilar; Boquete, Teresa; Pérez-Vázquez, Maria; Marín, Mercedes; Bouza, Emilio

    2009-01-01

    In a point-prevalence study performed in 145 Spanish hospitals in 2006, we collected 463 isolates of Staphylococcus aureus in a single day. Of these, 135 (29.2%) were methicillin (meticillin)-resistant S. aureus (MRSA) isolates. Susceptibility testing was performed by a microdilution method, and mecA was detected by PCR. The isolates were analyzed by pulsed-field gel electrophoresis (PFGE) after SmaI digestion, staphylococcal chromosomal cassette mec (SCCmec) typing, agr typing, spa typing wi...

  9. Determination of Enantiomeric Purity for Lactic Acid in Fermentation Broth by Rhizopus Oryzae with High Performance Liquid Chromatography%高效液相色谱法测定米根霉乳酸发酵液中乳酸的光学纯度

    Institute of Scientific and Technical Information of China (English)

    白冬梅; 赵学明; 胡宗定

    2001-01-01

    A procedure for the resolution of DL-lactic acid and th edeterminatio n of D-isomer ratio in L-lactic acid fermentation broth by Rhizopus oryzae is desc ribed. The effects of pH of mobile phase and concentration of chiral mobile pha s e additives on resolution of DL-lactic acid were investigated. The optical is om ers of lactic acid were resolved by RP-HPLC with 2,3,6-tri-O-β-cyc lode xtrin(TM-β-CD) as a chiral mobile phase additive, and C18 column as stationary phase, and detected at wavelength 210 nm. The results sh owed that a correction factor should be introduced into the equation for ca lc ulation of the percentage of D-lactic acid, because the UV absorption of D-lac tic acid and L-lactic acid might not be the same when TM-β-CD was presen t. Quantitation was achieved with external standard method, the average rec overy was 100.4%, and the relative standard deviation was 0.82%. This method can be used for the determination of the percentage of D-isomer in L-lactic a cid fermentation broth by Rhizopus oryzae, and it is simple, rapid and accur ate. T he results showed that the mass fraction of D-isomer in the fermented broth inc reased during the period of storage.%以2,3,6-三甲基-β-环糊精作手性流动相添加剂,C18柱为固定相,考察了流动相的pH和手性流动相添加剂浓度对乳酸对映体分离度的影响,建立了DL-乳酸的拆分定量分析方法。用归一化法分析DL-乳酸时引入了定量校正因子,确定了外标法测定L-乳酸的回归方程、精密度和回收率。测定了米根霉L-乳酸发酵液中D-型异构体的质量分数及其随放置时间的变化情况。

  10. Evaluation of Multiple Methods for Detection of Gastrointestinal Colonization of Carbapenem-Resistant Organisms from Rectal Swabs.

    Science.gov (United States)

    Simner, Patricia J; Martin, Isabella; Opene, Belita; Tamma, Pranita D; Carroll, Karen C; Milstone, Aaron M

    2016-06-01

    Rectal swabs from high-risk patients were screened for carbapenem-resistant organisms (CROs) using several methods. The direct MacConkey plate method was the most sensitive for CROs (95%), while chromID CARBA and the Check-Direct CPE screen assay were the most sensitive for the detection of carbapenemase-producing organisms (CPOs) (100%; all blaKPC). All methods had a specificity of >90% for CROs, and for CPOs, the specificity ranged from 85 to 98%. Broth enrichment methods performed poorly compared to direct inoculation methods, negating the need for the broth enrichment step. PMID:27053674

  11. Influence of carvacrol and 1,8-cineole on cell viability, membrane integrity, and morphology of Aeromonas hydrophila cultivated in a vegetable-based broth.

    Science.gov (United States)

    de Sousa, Jossana Pereira; de Oliveira, Kataryne Árabe Rimá; de Figueiredo, Regina Celia Bressan Queiroz; de Souza, Evandro Leite

    2015-02-01

    This study investigated the effects of carvacrol (CAR) and 1,8-cineole (CIN) alone (at the MIC) or in combination at subinhibitory amounts (both at 1/8 MIC) on the cell viability, membrane permeability, and morphology of Aeromonas hydrophila INCQS 7966 (A. hydrophila) cultivated in a vegetable-based broth. CAR and CIN alone or in combination severely affected the viability of the bacteria and caused dramatic changes in the cell membrane permeability, leading to cell death, as observed by confocal laser microscopy. Scanning and transmission electron microscopy images of bacterial cells exposed to CAR or CIN or the mixture of both compounds revealed severe changes in cell wall structure, rupture of the plasma membrane, shrinking of cells, condensation of cytoplasmic content, leakage of intracellular material, and cell collapse. These findings suggest that CAR and CIN alone or in combination at subinhibitory amounts could be applied to inhibit the growth of A. hydrophila in foods, particularly as sanitizing agents in vegetables. PMID:25710162

  12. Fermentative utilization of coffee mucilage using Bacillus coagulans and investigation of down-stream processing of fermentation broth for optically pure l(+)-lactic acid production.

    Science.gov (United States)

    Neu, Anna-Katrin; Pleissner, Daniel; Mehlmann, Kerstin; Schneider, Roland; Puerta-Quintero, Gloria Inés; Venus, Joachim

    2016-07-01

    In this study, mucilage, a residue from coffee production, was investigated as substrate in fermentative l(+)-lactic acid production. Mucilage was provided as liquid suspension consisting glucose, galactose, fructose, xylose and sucrose as free sugars (up to 60gL(-1)), and used directly as medium in Bacillus coagulans batch fermentations carried out at 2 and 50L scales. Using mucilage and 5gL(-1) yeast extract as additional nitrogen source, more than 40gL(-1) lactic acid was obtained. Productivity and yield were 4-5gL(-1)h(-1) and 0.70-0.77g lactic acid per g of free sugars, respectively, irrespective the scale. Similar yield was found when no yeast extract was supplied, the productivity, however, was 1.5gL(-1)h(-1). Down-stream processing of culture broth, including filtration, electrodialysis, ion exchange chromatography and distillation, resulted in a pure lactic acid formulation containing 930gL(-1)l(+)-lactic acid. Optical purity was 99.8%. PMID:27035470

  13. Efficacy of mini VIDAS for the detection of Campylobacter spp. from retail broiler meat enriched in Bolton broth, with or without the supplementation of blood.

    Science.gov (United States)

    Liu, Lin; Hussain, Syeda K; Miller, Robert S; Oyarzabal, Omar A

    2009-11-01

    The goals of this study were to evaluate the efficacy of the mini VIDAS automated immunoassay chemistry system to detect Campylobacter spp. from retail broiler meat enriched in Bolton broth supplemented with lysed blood (B+B) or without blood (B-B), and to detect positive samples at 24 versus 48 h after enrichment. Retail broiler meat was enriched and tested for Campylobacter spp. with the mini VIDAS and with an agar plate. Isolates were speciated with a multiplex PCR and typed with pulsed-field gel electrophoresis (PFGE) to evaluate relatedness of isolates collected from subsamples enriched in B+B or B-B. The number of Campylobacter-positive samples by mini VIDAS was similar (P > 0.05) to the results found with traditional plating media for naturally contaminated broiler meat, regardless of whether the comparison was made between B+B and B-B, or among different meat products (breast, tenders, and thighs). More positive samples were found at 48 h of enrichment than at 24 h of enrichment (P plate media, and retail broiler samples need to be enriched for 48 h to avoid false negatives. PMID:19903413

  14. A Novel Technique of Synthesis of Highly Fluorescent Carbon Nanoparticles from Broth Constituent and In-vivo Bioimaging of C. elegans.

    Science.gov (United States)

    Pramanik, A; Kole, A K; Krishnaraj, R N; Biswas, S; Tiwary, C S; Varalakshmi, P; Rai, S K; Kumar, B A; Kumbhakar, P

    2016-09-01

    Here we have demonstrated a novel single step technique of synthesis of highly fluorescent carbon nanoparticles (CNPs) from broth constituent and in vivo bioimaging of Caenorhabditis elegans (C. elegans) with the synthesized CNPs has been presented. The synthesized CNPs has been characterized by the UV-visible (UV-Vis) absorption spectroscopy, transmission electron microscopy (TEM) and Raman studies. The sp (2) cluster size of the synthesized samples has been determined from the measured Raman spectra by fitting it with the theoretical skew Lorentzian (Breit-Wigner- Fano (BWF)) line shape. The synthesised materials are showing excitation wavelength dependent tunable photoluminescence (PL) emission characteristics with a high quantum yield (QY) of 3 % at a very low concentration of CNPs. A remarkable increase in the intensity of PL emission from 16 % to 39 % in C. elegans has also been observed when the feeding concentration of CNPs to C. elegans is increased from 0.025 % to 0.1 % (w/v). The non-toxicity and water solubility of the synthesized material makes it ideal candidate for bioimaging. PMID:27380200

  15. Kinetics of biogas production from fermentation broth of wild cocoyam codigested with cow paunch in batch mode

    Directory of Open Access Journals (Sweden)

    I.C.E. Umeghalu

    2015-03-01

    Full Text Available Mathematical models are useful in solving the stability problems often exhibited by anaerobic digestion process. Kinetics of batch anaerobic digestion of cocoyam waste mixed with cow paunch for biogas production in batch mode was studied for 30 days hydraulic retention time (HRT. Data from cumulative biogas yield obtained during the experimental stages was fitted to C-NIKBRAN mathematical model based on first order reaction which adequately predicted the kinetic behavior of the substrate’s anaerobic biodegradability. The validity of the applied model was also verified through application of the regression model (ReG (Least Square Method using Excel Version 2003 in predicting the trend of the experimental results. Comparative analysis of Figs. 7-10 show very close alignment of curves which precisely translated into significantly similar trend of data point’s distribution for experimental (ExD, derived model (MoD and regression model-predicted (ReG results of cumulative biogas yield. Also, critical analysis of data obtained from experiment and derived model show low deviations on the part of the model-predicted values relative to values obtained from the experiment. This necessitated the introduction of correction factor, to bring the model-predicted cumulative biogas yield to those of the corresponding experimental values. Deviational analysis from strongly indicates that cumulative biogas yield was most reliable based on the associated admissible deviation of the model-predicted cumulative biogas yield from the corresponding experimental values; 9.2% within the pH range. The values of cumulative biogas yield within the highlighted deviation indicates over 90% confidence level for the applied model and over 0.9 effective dependency coefficients (EDC of cumulative biogas yield on pH, chemical oxygen demand (COD, total viable count (TVC and total dissolved solids (TDS. Also, deviation of model-predicted cumulative biogas yield from

  16. Interpretive criteria for mupirocin susceptibility testing of Staphylococcus spp. using CLSI guidelines.

    LENUS (Irish Health Repository)

    Creagh, S

    2012-02-03

    Mupirocin is an antimicrobial agent commonly used to treat staphylococcal infection or to eliminate persistent carriage. To date, interpretive criteria have not been established to define susceptibility or resistance when performing mupirocin susceptibility testing. In this evaluation, using CLSI guidelines, a total of 502 staphylococci comprising 219 methicillin-sensitive Staphylococcus aureus, 222 methicillin-resistant S. aureus and 61 coagulase-negative staphylococci are tested by broth microdilution, disc diffusion and E-test. Disc diffusion using 5 microg mupirocin discs was found to be a reliable method to distinguish susceptible and resistant strains. Minimum inhibitory concentration (MIC) determination was required to differentiate low-level and high-level resistance to mupirocin. E-test was found to be an accurate alternative to broth microdilution for the routine determination of MIC values of staphylococci to mupirocin. Broth microdilution and disc-diffusion results were plotted on a scattergram, and error rates were calculated. No errors were found using susceptibility criteria of < 4 microg\\/mL (MIC) and > 19 mm (zone diameter).

  17. Establishing the validity of different susceptibility testing methods to evaluate the in vitro activity of amoxicillin-clavulanate against Escherichia coli.

    Science.gov (United States)

    María, Díez-Aguilar; María-Isabel, Morosini; María-Carmen, Conejo; Álvaro, Pascual; Jorge, Calvo; Luis, Martínez-Martínez; Francesc, Marco; Jordi, Vila; Adriana, Ortega; Jesús, Oteo; Rafael, Cantón

    2016-04-01

    Amoxicillin-clavulanate MICs of 160 Escherichia coli isolates with characterized resistance mechanisms were obtained by 2 MIC gradient strip brands, 3 automated systems, and reference ISO microdilution method using EUCAST (fixed 2μg/mL clavulanate) and CLSI (2:1 ratio) criteria. Discrepancies, mainly obtained with gradient strips, lead to an essential agreement range of 76.2-92.5. PMID:26830054

  18. Escherichia coli K-12 survives anaerobic exposure at pH 2 without RpoS, Gad, or hydrogenases, but shows sensitivity to autoclaved broth products.

    Directory of Open Access Journals (Sweden)

    Daniel P Riggins

    Full Text Available Escherichia coli and other enteric bacteria survive exposure to extreme acid (pH 2 or lower in gastric fluid. Aerated cultures survive via regulons expressing glutamate decarboxylase (Gad, activated by RpoS, cyclopropane fatty acid synthase (Cfa and others. But extreme-acid survival is rarely tested under low oxygen, a condition found in the stomach and the intestinal tract. We observed survival of E. coli K-12 W3110 at pH 1.2-pH 2.0, conducting all manipulations (overnight culture at pH 5.5, extreme-acid exposure, dilution and plating in a glove box excluding oxygen (10% H2, 5% CO2, balance N2. With dissolved O2 concentrations maintained below 6 µM, survival at pH 2 required Cfa but did not require GadC, RpoS, or hydrogenases. Extreme-acid survival in broth (containing tryptone and yeast extract was diminished in media that had been autoclaved compared to media that had been filtered. The effect of autoclaved media on extreme-acid survival was most pronounced when oxygen was excluded. Exposure to H2O2 during extreme-acid treatment increased the death rate slightly for W3110 and to a greater extent for the rpoS deletion strain. Survival at pH 2 was increased in strains lacking the anaerobic regulator fnr. During anaerobic growth at pH 5.5, strains deleted for fnr showed enhanced transcription of acid-survival genes gadB, cfa, and hdeA, as well as catalase (katE. We show that E. coli cultured under oxygen exclusion (<6 µM O2 requires mechanisms different from those of aerated cultures. Extreme acid survival is more sensitive to autoclave products under oxygen exclusion.

  19. The influence of the cell free solution of lactic acid bacteria on tyramine production by food borne-pathogens in tyrosine decarboxylase broth.

    Science.gov (United States)

    Toy, Nurten; Özogul, Fatih; Özogul, Yesim

    2015-04-15

    The function of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on tyramine and other biogenic amine production by different food borne-pathogens (FBPs) was investigated in tyrosine decarboxylase broth (TDB) using HPLC. Cell free solutions were prepared from four LAB strains. Two different concentrations which were 50% (5 ml CFS+5 ml medium/1:1) and 25% (2.5 ml CFS+7.5 ml medium/1:3) CFS and the control without CFS were prepared. Both concentration of CFS of Streptococcus thermophilus and 50% CFS of Pediococcus acidophilus inhibited tyramine production up to 98% by Salmonella paratyphi A. Tyramine production by Escherichia coli was also inhibited by 50% CFS of Lactococcus lactis subsp. lactis and 25% CFS of Leuconostoc lactis. subsp. cremoris. The inhibitor effect of 50% CFS of P. acidophilus was the highest on tyramine production (55%) by Listeria monocytogenes, following Lc. lactis subsp. lactis and Leuconostoc mesenteroides subsp. cremoris (20%) whilst 25% CFS of Leu. mes. subsp. cremoris and Lc. lactis subsp. lactis showed stimulator effects (160%). The stimulation effects of 50% CFS of S. thermophilus and Lc. lactis subsp. lactis were more than 70% by Staphylococcus aureus comparing to the control. CFS of LAB strains showed statistically inhibitor effect since lactic acid inhibited microbial growth, decreased pH quickly and reduced the formation of AMN and BAs. Consequently, in order to avoid the formation of high concentrations of biogenic amines in fermented food by bacteria, it is advisable to use CFS for food and food products. PMID:25465993

  20. HPLC法检测丁二酸发酵液中的有机酸和水溶性维生素%Analysis of organic acids and hydrosoluble vitamins in succinic acid fernmentation broth by high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    赵文君; 屠春燕; 马海宁; 姜珉; 韦萍

    2011-01-01

    An innovative system of RP-HPLC method was proposed and optimized for detecting the organic acids and hydrosoluble vitamins in succinic acid fermentation broth. Under the optimal conditions, a Hedera ODS-2column (5μm, 4.6 mm × 250 mm) was used, the detection wavelength was 210 nm. Flow rate was 1.0 mL/min; Mobile phase A:20 mmol/L sodium dihydrogen phosphate(pH2.9), Mobile phase B: ACN; the gradient protocol was applied. All compounds were completely eluted in 15 min. All the correlation coefficients (r) of regression equations were above 0.9989, and the recoveries ranged from 92.5% to 100.3%. The relative standard deviations were in the range from 0.9% to 1.9%. By this method, organic acids and vitamins can be easily, rapidly and effectively determined.%采用高效液相色谱法(HPLC)同时检测丁二酸发酵液中的有机酸和水溶性维生素.以RP-HPLC为分离模式,选用Hedera ODS-2色谱柱(5μm,4.6×250mm),20mmol/L的磷酸盐缓冲溶液(pH 2.9)和乙腈为流动相,梯度洗脱,检测波长为210nm,建立了同时测定分析8种水溶性维生素和3种有机酸的方法,15min内所有组分都洗脱完全.8种水溶性维生素和3种有机酸在线性范围内峰面积与浓度呈良好的线性关系,相关系数为0.9989~0.9997;加标回收率在92.5%~100.3%之间;RSD(n=5):0.9%~1.9%.本方法采用同一种流动相既可以测定发酵液中的有机酸又可以测定水溶性维生素的含量.

  1. 超声波微波萃取-高效液相色谱法检测肉汤中的罂粟碱%Determination on papaverine in meat broth by Ultrasonic-microwave synergistic extraction-HPLC

    Institute of Scientific and Technical Information of China (English)

    信海红; 王庭欣; 朱路甲; 张晓宇; 王羽

    2012-01-01

    罂粟壳通常作为一种违法添加剂应用于火锅底料,目的是使人吃了上瘾,罂粟壳中含有多种生物碱,溶于肉汤食用后有害于健康.用75%的乙醇作为罂粟碱的提取液,利用超声波微波萃取仪对样品进行提取,高效液相色谱法(HPLC)进行含量分析,建立测定肉汤中罂粟碱的超声波微波萃取-高效液相色谱方法.其结果此方法罂粟碱在0.2~10mg/L范围内呈良好的线性关系,相关系数为0.9999.其平均回收率为94.47%;精密度相对标准偏差(RSD)为0.029%;重现性相对标准偏差(RSD)为0.665%.本方法具有回收率高、灵敏度高、精密度高等优点,可适用于食品中罂粟壳生物碱的检测,为确保农产品餐桌质量安全监控提供技术保障.%Pericarpium Papaverisis an illegal additives and usually used as food additives in the hot pot to let people addicted to the dish. Pericarpium Papaveris reacts with many typesof alkaloids in the dish and is harmful to healthA method for the determination of papaverine in meat broth using Ultrasonic - microwave synergistic extraction - HPLC was developed. The Ultrasonic- microwave synergistic extraction was used to extract the sample. In the concentration of 0. 2mg/L to 10mg/L, the result was linear (r = 0. 9999) . The average recovery was 94. 47% and the RSD of the precision was 0. 029% and the RSD of the reproducibility was 0. 665% . The method had high recovery, sensitivity and is very precise. It is suitable for the determination of papaverine in food. It provides the technical support to monitor the safety of agricultural products.

  2. Effectiveness of a Simplified Method for Isolation of Burkholderia pseudomallei from Soil

    OpenAIRE

    Limmathurotsakul, Direk; Wuthiekanun, Vanaporn; Amornchai, Premjit; Wongsuwan, Gumphol; Day, Nicholas P. J.; Peacock, Sharon J.

    2012-01-01

    Detection of environmental Burkholderia pseudomallei indicates a risk for melioidosis and is important for the development of a global risk map. We describe a simple method for detecting B. pseudomallei using direct culture of soil in enrichment broth. This gives a rate of positivity comparable to that obtained with a standard method but is cheaper and labor saving.

  3. The probability of growth of Listeria monocytogenes in cooked salmon and tryptic soy broth as affected by salt, smoke compound, and storage temperature.

    Science.gov (United States)

    Hwang, Cheng-An

    2009-05-01

    The objectives of this study were to examine and model the probability of growth of Listeria monocytogenes in cooked salmon containing salt and smoke (phenol) compound and stored at various temperatures. A growth probability model was developed, and the model was compared to a model developed from tryptic soy broth (TSB) to assess the possibility of using TSB as a substitute for salmon. A 6-strain mixture of L. monocytogenes was inoculated into minced cooked salmon and TSB containing 0-10% NaCl and 0-34 ppm phenol to levels of 10(2-3) cfu/g, and the samples were vacuum-packed and stored at 0--25 degrees C for up to 42 days. A total 32 treatments, each with 16 samples, selected by central composite designs were tested. A logistic regression was used to model the probability of growth of L. monocytogenes as a function of concentrations of salt and phenol, and storage temperature. Resulted models showed that the probabilities of growth of L. monocytogenes in both salmon and TSB decreased when the salt and/or phenol concentrations increased, and at lower storage temperatures. In general, the growth probabilities of L. monocytogenes were affected more profoundly by salt and storage temperature than by phenol. The growth probabilities of L. monocytogenes estimated by the TSB model were higher than those by the salmon model at the same salt/phenol concentrations and storage temperatures. The growth probabilities predicted by the salmon and TSB models were comparable at higher storage temperatures, indicating the potential use of TSB as a model system to substitute salmon in studying the growth behavior of L. monocytogenes may only be suitable when the temperatures of interest are in higher storage temperatures (e.g., >12 degrees C). The model for salmon demonstrated the effects of salt, phenol, and storage temperature and their interactions on the growth probabilities of L. monocytogenes, and may be used to determine the growth probability of L. monocytogenes in smoked

  4. Simple method for quantifying viable bacterial numbers in sputum.

    OpenAIRE

    Pye, A.; Stockley, R A; Hill, S. L.

    1995-01-01

    AIMS--To establish a simple method of quantitative culture for determining the viable bacterial numbers present in expectorated sputum samples. METHODS--Sputum samples were homogenised with dithiothreitol, sterile saline or glass beads to determine which method recovered the greatest number of viable bacteria. Culture broths were also incubated with dithiothreitol and sampled over time to determine its effect on bacterial viability. Sputum samples homogenised with dithiothreitol were diluted ...

  5. Method

    Directory of Open Access Journals (Sweden)

    Xixin Wang

    2012-01-01

    Full Text Available ZrO2 nanotube arrays were prepared by anodization method in aqueous electrolyte containing (NH42SO4 and NH4F. The morphology and structure of nanotube arrays were characterized through scanning electron microscope, X-ray diffraction, and infrared spectra analysis. The zirconia nanotube arrays were used as catalyst in esterification reaction. The effects of calcination temperature and electrolyte concentration on catalytic esterification activity have been investigated in detail. Experiments indicate that nanotube arrays have highest catalytic activity when the concentration of (NH42SO4 is 1 mol/L, the concentration of NH4F is 1 wt%, and the calcination temperature is 400°C. Esterification reaction yield of as much as 97% could be obtained under optimal conditions.

  6. An easy method for detection of nasopharyngeal carriage of multiple Streptococcus pneumoniae serotypes

    DEFF Research Database (Denmark)

    Kaltoft, Margit S.; Sørensen, Uffe; Slotved, Hans-Christian;

    2008-01-01

    In this paper, a simplified method for detection of pneumococcal carriage and for revealing the presence of several serotypes in a nasopharyngeal sample is evaluated. Enrichment broth was used for transportation and for the initial culturing of samples. All specimens were examined directly by the...

  7. Distribution of Listeria spp. in confectioners' pastries from western France: comparison of enrichment methods.

    Science.gov (United States)

    Ferron, P; Michard, J

    1993-06-01

    Three hundred samples of pastry from 100 different suppliers in western France, including butter-cream, whipped dairy cream and custard filled cakes from each supplier, were collected and tested for the occurrence of Listeria spp. in 25 g samples. Listeria spp. were detected in 21.7% of he samples: Listeria monocytogenes in 13.7%, Listeria innocua in 10% and Listeria seeligeri in 2.3%. Thirteen samples were contaminated with two species simultaneously. The frequency of contaminated samples was not related to the composition of the pastry filling used, but it seemed to increase with the number of aerobic contaminant microorganisms in the dairy cream-based samples. The contamination rate was dependent on the place of manufacture. The numbers of Listeria spp. and Listeria monocytogenes were estimated on positive samples at the 25 g level as follows: < 0.3/g, Listeria spp. in 47 samples, L monocytogenes in 27; 0.3-30/g, Listeria spp. in 13, L. monocytogenes in nine; 30-300/g, L. monocytogenes in one; 300-3000/g; L. monocytogenes in three; 700,000/g, L. monocytogenes in one. Various detection methods were tested, including two enrichments broths tested in parallel: a modified LEB broth using 10 mg/l acriflavine-HCl and the UVM 1 broth, with incubation at 30 degrees C and streaking onto PALCAM agar. The enrichment procedures were: (a) primary enrichment of 25 g sample and plating after 48 h and 7 days; (b) secondary enrichment by subculturing the primary enrichment broths incubated for 24 h and 6 days, into fresh enrichment broth, then plating after 24 h incubation; (c) pre-enrichment of 25 g sample for 24 h in the basal enrichment broths without inhibitors, followed by subculturing in complete broths which were plated after 24 h and 6 days incubation. In all cases, UVM performed better than the LEB broth. It was unnecessary to extend the primary enrichment period beyond 48 h. Secondary enrichments inoculated from 24-h incubated primary enrichments gave a slightly better

  8. Clinical and microbiological evaluation of four culture methods for the diagnosis of peritonitis in patients on continuous ambulatory peritoneal dialysis.

    OpenAIRE

    Doyle, P W; Crichton, E. P.; Mathias, R G; Werb, R

    1989-01-01

    A prospective study was performed to evaluate four culture methods for the diagnosis of bacterial peritonitis in patients on continuous ambulatory peritoneal dialysis. Peritonitis was present in 44 of 85 patient admissions (52%). The overall sensitivity of the culture methods in detecting peritonitis was 66%. The sensitivities of the individual methods were as follows: bag culture method, 61%; blood culture broth method, 51%; filter method, 54%; and plate method, 39%. Our broad definition of ...

  9. The concentration-dependent nature of in vitro amphotericin B-itraconazole interaction against Aspergillus fumigatus: isobolographic and response surface analysis of complex pharmacodynamic interactions.

    NARCIS (Netherlands)

    Meletiadis, J.; Dorsthorst, D.T.A. te; Verweij, P.E.

    2006-01-01

    The interaction between polyenes and azoles is not well understood. We therefore explored the in vitro combination of amphotericin B with itraconazole against 14 clinical Aspergillus fumigatus isolates (9 itraconazole susceptible and 5 itraconazole resistant) with a colorimetric broth microdilution

  10. In vitro activities of voriconazole (UK-109,496) against fluconazole-susceptible and -resistant Candida albicans isolates from oral cavities of patients with human immunodeficiency virus infection.

    OpenAIRE

    Ruhnke, M.; Schmidt-Westhausen, A; Trautmann, M

    1997-01-01

    The susceptibility of Candida albicans to a new antifungal triazole, voriconazole (UK-109,496), was investigated in 105 isolates obtained from the oral cavities of patients with human immunodeficiency virus (HIV) infection to study this drug's activity against fluconazole-susceptible and -resistant isolates. MICs were determined by a broth microdilution technique according to document M27-T from the National Committee for Clinical Laboratory Standards and by using a broth microdilution techni...

  11. Standardization of a quantification method for Salmonella spp. and Shigella spp. in specific liquid media

    Directory of Open Access Journals (Sweden)

    Sandra Patricia Rivera

    2010-09-01

    Full Text Available Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs have yielded promising results. Escherichia coli (E. coli is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella  spp. and Salmonella  spp. Additionally, the viable non-culturable (VNC state yields bacteria which are not detectable on many culture media. Objective: The main objective is to standardize a method for counting Salmonella  spp. and Shigella  spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO. Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella  spp. and Shigella  spp. Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth. Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the

  12. Standardization of a quantification method for Salmonella spp. and Shigella spp. in specific liquid media

    Directory of Open Access Journals (Sweden)

    Sandra Patricia Rivera

    2010-03-01

    Full Text Available Introduction: Chlorination is the most widely used disinfection process for drinking water production. The formation of chlorination carcinogenic by-products and chlorine intoxication by direct manipulation in small communities has motivated the study of alternative disinfection processes. In this sense, processes of advanced oxidation (PAOs have yielded promising results. Escherichia coli (E. coli is customarily used as faecal bacterial indicator to determine the efficiency of disinfection processes. However, it has been shown that E. coli is less resistant to disinfection than other enteric bacteria such as Shigella spp. and Salmonella spp. Additionally, the viable non-culturable (VNC state yields bacteria which are not detectable on many culture media.Objective: The main objective is to standardize a method for counting Salmonella spp. and Shigella spp. in specific liquid media to reliably quantify the bacteriological potential risk related to disinfection processes based on PAO.Methods: The study followed a randomized bi-factorial experimental design and the Duncan multiple comparison test. This design allowed the selection of specific liquid media to fittingly standardize the counting of Salmonella spp. and Shigella spp.Results: We found that the best broth for counting Salmonella typhimurium strain at different concentrations in pure and mixed cultures was the Rappaport broth RP, the EE broth also allowed growing the two bacterial species tested in this research. Nonetheless, the latter results suggest the use of additional tests for this particular broth.Discussion: There was a variation in the counting results when pure cultures were used compared to those obtained from mixtures of microorganisms. It was also noted that Salmonella typhimurium and Shigella sonnei, were recovered from minimal concentrations in both RP and EE broths, respectively. To some extent, this suggests an additional confirmative method when using the EE® broth

  13. Methods for increasing the production of ethanol from microbial fermentation

    Science.gov (United States)

    Gaddy, James L.; Arora, Dinesh K.; Ko, Ching-Whan; Phillips, John Randall; Basu, Rahul; Wikstrom, Carl V.; Clausen, Edgar C.

    2007-10-23

    A stable continuous method for producing ethanol from the anaerobic bacterial fermentation of a gaseous substrate containing at least one reducing gas involves culturing a fermentation bioreactor anaerobic, acetogenic bacteria in a liquid nutrient medium; supplying the gaseous substrate to the bioreactor; and manipulating the bacteria in the bioreactor by reducing the redox potential, or increasing the NAD(P)H TO NAD(P) ratio, in the fermentation broth after the bacteria achieves a steady state and stable cell concentration in the bioreactor. The free acetic acid concentration in the bioreactor is maintained at less than 5 g/L free acid. This method allows ethanol to be produced in the fermentation broth in the bioreactor at a productivity greater than 10 g/L per day. Both ethanol and acetate are produced in a ratio of ethanol to acetate ranging from 1:1 to 20:1.

  14. Novel Method of Lactic Acid Production by Electrodialysis Fermentation

    OpenAIRE

    Hongo, Motoyoshi; Nomura, Yoshiyuki; Iwahara, Masayoshi

    1986-01-01

    In lactic acid fermentation by Lactobacillus delbrueckii, the produced lactic acid affected the lactic acid productivity. Therefore, for the purpose of alleviating this inhibitory effect, an electrodialysis fermentation method which can continuously remove produced lactic acid from the fermentation broth was applied to this fermentation process. As a result, the continuation of fermentation activity was obtained, and the productivity was three times higher than in non-pH-controlled fermentati...

  15. Methods for improved recovery of Listeria monocytogenes from cheese.

    OpenAIRE

    Yousef, A. E.; Ryser, E T; Marth, E. H.

    1988-01-01

    Method of homogenization (Waring blender versus stomacher), type of diluent (tryptose broth [TB] versus aqueous 2% trisodium citrate), and temperature of diluent (20 versus 40 degrees C) were compared for recovery of Listeria monocytogenes from freshly made and ripened Colby cheese. By using direct plating on McBride listeria agar, significantly higher numbers of L. monocytogenes were recovered when cheese samples were (i) homogenized for 2 min with the blender rather than the stomacher (P le...

  16. A new spectrophotometric method for quantification of potassium solubilized by bacterial cultures.

    Science.gov (United States)

    Rajawat, Mahendra Vikram Singh; Singh, Surender; Saxena, Anil Kumar

    2014-03-01

    A new spectrophotometric method was developed for the quantification of potassium in the culture broth supernatant of K-solubilizing bacteria. The standard curve of potassium with the new method, which is based on the measurement of cobalt, showed a regression coefficient (R2) of 0.998. The quantification values of potassium obtained with flame photometric method and the newly developed method showed a significant correlation (r) of 0.978. The new method depends on the precipitation of sodium cobaltinitrite with solubilized potassium in liquid medium as potassium sodium cobaltinitrite, which develops bluish green colour by the addition of conc. HCl. The intensity of developed colour can be recorded at 623 nm. This method involves less number of steps, is easy and time saving, and can be used for the reliable estimation of available potassium in culture broth supernatant of K-solubilizing bacteria. PMID:24669669

  17. EFFICIENCY OF THE ENRICHMENT BROTHES (LEB1 AND LEB2) AND SELECTIVE AGARS (LPM AND LSAB-CAN) TO ISOLATE BACTERIA OF THE GENUS Listeria IN MEAT AND RESIDUAL WATER OF WASHING CARCASS EFICIÊNCIA DOS CALDOS DE ENRIQUECIMENTO (LEB1 E LEB2) E DOS ÁGARES SELETIVOS (LPM E LSAB-CAN) NO ISOLAMENTO DE BACTÉRIAS DO GÊNERO Listeria EM CARNE BOVINA E ÁGUA RESIDUÁRIA DE LAVAGEM DE CARCAÇA

    OpenAIRE

    Albenones José de Mesquita; Sebastião Timo Iaria; Iolanda Aparecida Nunes

    2007-01-01

    In this paper it was observed the efficiency of enrichment brothes for Listeria (LEB1 and LEB2), associated to the passage of the culture through a 0.25% potassium hidroxide solution, verifying that the secondary enrichment broth (LEB2 and LEB2KOH) was better than the primary enrichment broth (LEB1KOH) for this purpose. On the other hand, lithium chloride-phenylethanol-moxalactam agar and the s...

  18. QUANTIFICATION OF LISTERIA MONOCYTOGENES IN MILK BY MPN-PCR AND MPN-CULTURE METHODS

    OpenAIRE

    Mahzad Hosseini; Abdollah Jamsidi; Saeid Khanzadi

    2014-01-01

    The aim of this study was to compare the MPN-PCR (Most Probable Number- Polymerase Chain Reaction) and MPN-Culture methods in enumerating of Listeria monocytogenes in milk. In order to compare the accuracy of these methods, 103 cell/ml Listeria monocytogenes and different background bacteria which may be present in raw milk, were inoculated in sterilized milk. After preparing serial dilutions, three replicates per dilution were inoculated in tubes containing listeria enrichment broth. After ...

  19. Isolation of Mycobacterium kumamotonense from a patient with pulmonary infection and latent tuberculosis.

    Science.gov (United States)

    Kontos, Fanourios; Mavromanolakis, Dimitrios Nikitas; Zande, Marina Chari; Gitti, Zoe Georgios

    2016-01-01

    Mycobacterium kumamotonense is a novel, slow-growing non-chromogenic nontuberculous mycobacterium, which belongs to Mycobacterium terrae complex. We report, for the first time in Greece, the isolation of M. kumamotonense from an immunocompetent patient with pulmonary infection and latent tuberculosis. M. kumamotonense was identified by sequencing analysis of 16S rDNA and 65-kDa heat shock protein genes while by commercial molecular assays it was misidentified as Mycobacterium celatum. Antibiotic susceptibility testing was performed by the reference broth microdilution method. The strain was susceptible to amikacin, clarithromycin, rifampin, ciprofloxacin, moxifloxacin, rifabutin, ethambutol and linezolid. PMID:27080783

  20. In Vitro Activity of Colistin (Polymyxin E) against 3,480 Isolates of Gram-Negative Bacilli Obtained from Patients in Canadian Hospitals in the CANWARD Study, 2007-2008▿

    OpenAIRE

    Walkty, A.; DeCorby, M.; Nichol, K; Karlowsky, J A; Hoban, D. J.; Zhanel, G G

    2009-01-01

    The in vitro activity of colistin was evaluated versus 3,480 isolates of gram-negative bacilli using CLSI broth microdilution methods. The MIC90 of colistin was ≤2 μg/ml against a variety of clinically important gram-negative bacilli, including Escherichia coli, Klebsiella spp., Enterobacter spp., Acinetobacter baumannii, and Pseudomonas aeruginosa. All multidrug-resistant (n = 76) P. aeruginosa isolates were susceptible to colistin (MIC, ≤2 μg/ml). These data support a role for colistin in t...

  1. Antifungal activity of raw extract and flavanons isolated from Piper ecuadorense from Ecuador

    OpenAIRE

    Jorge Ramirez; Luis Cartuche; Vladimir Morocho; Silvio Aguilar; Omar Malagon

    2013-01-01

    The MeOH extract of Piper ecuadorense Sodiro, Piperaceae, was chosen for metabolite isolation and elucidation due to the strong antifungal activity exhibited, measured by means of the broth microdilution method. Two known flavonoids: pinostrobin (1) and pinocembrin (2) were isolated from 4.16 g. of dichloromethane extract by column chromatography, using a gradient of hexane/EtOAc. A total of 20 mg of 1 were obtained from the fraction eluted with hexane-EtOAc 95:5 v/v, and 100 mg of 2 were obt...

  2. Differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis

    Science.gov (United States)

    Maboni, Grazieli; Gressler, Leticia T.; Espindola, Julia P.; Schwab, Marcelo; Tasca, Caiane; Potter, Luciana; de Vargas, Agueda Castagna

    2015-01-01

    The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented. PMID:26273272

  3. Lanthanum triflate triggered synthesis of tetrahydroquinazolinone derivatives of N-allyl quinolone and their biological assessment

    Directory of Open Access Journals (Sweden)

    Jardosh Hardik H.

    2012-01-01

    Full Text Available A series of 24 derivatives of tetrahydroquinazolinone has been synthesized by one-pot cyclocondensation reaction of N-allyl quinolones, cyclic β-diketones and (thiourea/N-phenylthiourea in presence of lanthanum triflate catalyst. This methodology allowed us to achieve the products in excellent yield by stirring at room temperature. All the synthesized compounds were investigated against a representative panel of pathogenic strains using broth microdilution MIC (minimum inhibitory concentration method for their in vitro antimicrobial activity. Amongst these sets of heterocyclic compounds 5h, 6b, 6h, 5f, 5l, 5n and 6g found to have admirable activity.

  4. 高效液相色谱法同时测定发酵液中木糖醇和L-木酮糖的含量%Simultaneous determination of xylitol and L-xylulose in fermentation broth with high performance liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    葛驰宇; 蒋洁; 翁旭; 张君丽

    2016-01-01

    目的:建立高效液相色谱( HPLC)同时测定发酵液中底物木糖醇和产物L-木酮糖含量的方法。方法采用C18色谱柱(250 mm ×4.6 mm),柱温35℃,以乙腈-水(体积比为85∶15)为流动相,流速0.8 mL/min。用示差折光检测器检测木糖醇,检测器温度为33℃。用紫外检测器在室温下检测L-木酮糖,检测波长为210 nm。结果所得木糖醇的线性范围为0.50~30.00 g/L,相关系数为0.9995,最低检出限为0.18 g/L,最低定量限为0.58 g/L;所得L-木酮糖的线性范围为0.30~30.00 g/L,相关系数为0.9986,最低检出限0.15 g/L,最低定量限为0.40 g/L;木糖醇的日内和日间相对标准偏差( RSD)均分别小于0.64%和0.80%,L-木酮糖的日内和日间RSD均分别小于0.31%和0.59%;回收率均在99.00%~101.00%之间。结论建立的HPLC法不受发酵液中其他组分的干扰,可同时测定底物木糖醇和产物L-木酮糖的含量,并实时监控发酵的全过程。%Objective A high performance liquid chromatographic (HPLC) method was established for the simultaneous determination of xylitol and L-xylulose in fermentation broth.Methods The chromatographic conditions were as follows:C18 column (250 mm ×4.6 mm) with the temperature 35℃, acetonitrile-water (85∶15,v/v)as mobile phase with the flow rate of 0.8 mL/min.Xylitol was detected by refractive index (RI) detector at 33℃and L-xylulose was determined by ultraviolet ( UV) detector at 210 nm at room temperature.Results This method showed good linearity over the range from 0.50~30.00 g/L with a correlation coefficient of 0.9995 for xylitol and 0.30~30.00 g/L with a correlation coefficient of 0.9986 for L-xylulose. Moreover, the limit of quantification (LOQ) for xylitol and L-xylulose were 0.58 and 0.40,respectively.The limit of determination (LOD) for xylitol and L-xylulose were 0.18 and 0

  5. Decreasing the Level of Ethyl Acetate in Ethanolic Fermentation Broths of Escherichia coli KO11 by Expression of Pseudomonas putida estZ Esterase†

    OpenAIRE

    Hasona, Adnan; York, S W; Yomano, L. P.; Ingram, L O; Shanmugam, K T

    2002-01-01

    During the fermentation of sugars to ethanol relatively high levels of an undesirable coproduct, ethyl acetate, are also produced. With ethanologenic Escherichia coli strain KO11 as the biocatalyst, the level of ethyl acetate in beer containing 4.8% ethanol was 192 mg liter−1. Although the E. coli genome encodes several proteins with esterase activity, neither wild-type strains nor KO11 contained significant ethyl acetate esterase activity. A simple method was developed to rapidly screen bact...

  6. Comparison of different sampling techniques and of different culture methods for detection of group B streptococcus carriage in pregnant women

    Directory of Open Access Journals (Sweden)

    Verhelst Rita

    2010-09-01

    Full Text Available Abstract Background Streptococcus agalactiae (group B streptococcus; GBS is a significant cause of perinatal and neonatal infections worldwide. To detect GBS colonization in pregnant women, the CDC recommends isolation of the bacterium from vaginal and anorectal swab samples by growth in a selective enrichment medium, such as Lim broth (Todd-Hewitt broth supplemented with selective antibiotics, followed by subculture on sheep blood agar. However, this procedure may require 48 h to complete. We compared different sampling and culture techniques for the detection of GBS. Methods A total of 300 swabs was taken from 100 pregnant women at 35-37 weeks of gestation. For each subject, one rectovaginal, one vaginal and one rectal ESwab were collected. Plating onto Columbia CNA agar (CNA, group B streptococcus differential agar (GBSDA (Granada Medium and chromID Strepto B agar (CA, with and without Lim broth enrichment, were compared. The isolates were confirmed as S. agalactiae using the CAMP test on blood agar and by molecular identification with tDNA-PCR or by 16S rRNA gene sequence determination. Results The overall GBS colonization rate was 22%. GBS positivity for rectovaginal sampling (100% was significantly higher than detection on the basis of vaginal sampling (50%, but not significantly higher than for rectal sampling (82%. Direct plating of the rectovaginal swab on CNA, GBSDA and CA resulted in detection of 59, 91 and 95% of the carriers, respectively, whereas subculturing of Lim broth yielded 77, 95 and 100% positivity, respectively. Lim broth enrichment enabled the detection of only one additional GBS positive subject. There was no significant difference between GBSDA and CA, whereas both were more sensitive than CNA. Direct culture onto GBSDA or CA (91 and 95% detected more carriers than Lim broth enrichment and subculture onto CNA (77%. One false negative isolate was observed on GBSDA, and three false positives on CA. Conclusions In

  7. Direct MALDI-TOF Mass Spectrometry Assay of Blood Culture Broths for Rapid Identification of Candida Species Causing Bloodstream Infections: an Observational Study in Two Large Microbiology Laboratories

    OpenAIRE

    Spanu, Teresa; Posteraro, Brunella; Fiori, Barbara; D'Inzeo, Tiziana; Campoli, Serena; Ruggeri, Alberto; Tumbarello, Mario; Canu, Giulia; Trecarichi, Enrico Maria; Parisi, Gabriella; Tronci, Mirella; Sanguinetti, Maurizio; Fadda, Giovanni

    2012-01-01

    We evaluated the reliability of the Bruker Daltonik's MALDI Biotyper system in species-level identification of yeasts directly from blood culture bottles. Identification results were concordant with those of the conventional culture-based method for 95.9% of Candida albicans (187/195) and 86.5% of non-albicans Candida species (128/148). Results were available in 30 min (median), suggesting that this approach is a reliable, time-saving tool for routine identification of Candida species causing...

  8. Quantifying antimicrobial resistance at veal calf farms.

    Directory of Open Access Journals (Sweden)

    Angela B Bosman

    Full Text Available This study was performed to determine a sampling strategy to quantify the prevalence of antimicrobial resistance on veal calf farms, based on the variation in antimicrobial resistance within and between calves on five farms. Faecal samples from 50 healthy calves (10 calves/farm were collected. From each individual sample and one pooled faecal sample per farm, 90 selected Escherichia coli isolates were tested for their resistance against 25 mg/L amoxicillin, 25 mg/L tetracycline, 0.5 mg/L cefotaxime, 0.125 mg/L ciprofloxacin and 8/152 mg/L trimethoprim/sulfamethoxazole (tmp/s by replica plating. From each faecal sample another 10 selected E. coli isolates were tested for their resistance by broth microdilution as a reference. Logistic regression analysis was performed to compare the odds of testing an isolate resistant between both test methods (replica plating vs. broth microdilution and to evaluate the effect of pooling faecal samples. Bootstrap analysis was used to investigate the precision of the estimated prevalence of resistance to each antimicrobial obtained by several simulated sampling strategies. Replica plating showed similar odds of E. coli isolates tested resistant compared to broth microdilution, except for ciprofloxacin (OR 0.29, p ≤ 0.05. Pooled samples showed in general lower odds of an isolate being resistant compared to individual samples, although these differences were not significant. Bootstrap analysis showed that within each antimicrobial the various compositions of a pooled sample provided consistent estimates for the mean proportion of resistant isolates. Sampling strategies should be based on the variation in resistance among isolates within faecal samples and between faecal samples, which may vary by antimicrobial. In our study, the optimal sampling strategy from the perspective of precision of the estimated levels of resistance and practicality consists of a pooled faecal sample from 20 individual animals, of which

  9. Beneficial effects of the ethanol extract from the dry matter of a culture broth of Inonotus obliquus in submerged culture on the antioxidant defence system and regeneration of pancreatic beta-cells in experimental diabetes in mice.

    Science.gov (United States)

    Xu, Hong-Yu; Sun, Jun-En; Lu, Zhen-Ming; Zhang, Xiao-Mei; Dou, Wen-Fang; Xu, Zheng-Hong

    2010-04-01

    The antihyperglycaemic and antilipidperoxidative effects of the ethanol extract from the dry matter of a culture broth (DMCB) of Inonotus obliquus were investigated in alloxan-induced diabetic mice and the possible mechanism of action was also discussed. In alloxan-induced diabetic mice, treatment with the ethanol extract from DMCB of I. obliquus (30 and 60 mg kg(-1) body weight (b.w.) for 21 days) showed a significant decrease in blood glucose level: the percentage reductions on the 7th day were 11.54 and 11.15%, respectively. However, feeding of this drug for three weeks produced reduction of 22.51 and 24.32%. Furthermore, the ethanol extract from the DMCB of I. obliquus treatment significantly decreased serum contents of free fatty acids, total cholesterol, triglycerides and low-density lipoprotein-cholesterol, whereas it effectively increased high-density lipoprotein-cholesterol, insulin levels and hepatic glycogen contents in livers of diabetic mice. Besides this, the ethanol extracts from the DMCB treatment significantly increased catalase, superoxide dismutase and glutathione peroxidase activities, except for decreasing the maleic dialdehyde level in diabetic mice. Histological morphology examination showed that the ethanol extract from the DMCB of I. obliquus restored the damage of pancreatic tissues in mice with diabetes mellitus. The results showed that the ethanol extract from the DMCB of I. obliquus possesses significant antihyperglycaemic, antilipidperoxidative and antioxidant effects in alloxan-induced diabetic mice. PMID:20397104

  10. Diferentes condimentos vegetais: avaliação sensorial e de atividade antibacteriana em preparação alimentar com frango cozido Different spice plants: sensorial evaluation and antibacterial activity in chicken broth

    Directory of Open Access Journals (Sweden)

    F. Rodrigues

    2011-01-01

    , individualmente, atividade antibacteriana significativa, mesmo que sem significância quando comparados entre si. Contudo, em relação ao tempo de início da atividade antibacteriana, destacou-se a pimenta dedo-de-moça, enquanto que, em relação ao prolongamento dessa ação no tempo, destacou-se o alho nirá. As 12 plantas condimentares em estudo tiveram atestada a sensorialidade, sendo que as quatro plantas com destaque tiveram a atividade anti-coliforme termo-resistente comprovada in loco. Diferentes condimentos vegetais foram capazes de fornecer qualificação sensorial e sanitária em caldo com frango cozido, em condições domésticas de manuseio.Based on the in vitro antibacterial activity predetermined for 12 spice plants with ethnographic indicator, this feature was tested in loco in the model cooked chicken broth. First, ten evaluators were trained, according to the current legislation for Free and Informed Consent, providing previous knowledge about the plants parsley (Petroselinum sativum, marjoram (Origanum X aplii and Origanum majorana, basil (Ocimum basilicum, common sage (Salvia officinalis, thyme (Thymus vulgaris, anis-like spice (Ocimum selloi, african basilicum (Ocimum gratissimum, nirá garlic (Allium tuberosum, leek (Allium porrum, turmeric (Curcuma longa and "dedo-de-moça" chili (Capsicum baccatum. Those spices were individually added to the chicken broth to perform a Hedonic Scale-like Acceptance Test, selecting four of the twelve spices that had higher sensory acceptance, "dedo-de-moça" chili, nirá garlic, leek and thyme. A new Acceptance Test was then performed using low, medium and high concentrations of those four spices to establish the most acceptable sensory intensities. The elected quantities (0.5 g "dedo-de-moça" chili, 15 g nirá garlic, 15 g leek and 5 g thyme were added to the chicken broth, then challenged with Escherichia coli (ATCC 11229 at a final 10 concentration of CFU/mL, the tolerated limit according to legislation. The control

  11. Antifungal activity of ibuprofen against aspergillus species and its interaction with common antifungal drugs

    Institute of Scientific and Technical Information of China (English)

    LI Li-juan; CHEN Wei; XU Hui; WAN Zhe; LI Ruo-yu; LIU Wei

    2010-01-01

    Background The incidence of invasive aspergillosis (IA) has increased in frequency in immunocompromised patients with a variety of diseases. The poor prognosis might be due to limited treatment option. This study aimed to evaluate antifungal activity of ibuprofen against clinical isolates of aspergillus species, as well as its interaction with azoles or with amphotericin B or with micafungin.Methods Antifungal activity of ibuprofen against 10 strains of Aspergillus fumigatus, Aspergillus flavus, and Aspergillus terreus were tested with both disk diffusion assay and standard broth microdilution method. To determine whether ibuprofen combined with itraconazole, voriconazole, amphotericin B, or micafungin had interactive effects on aspergillus spp., we used both disk diffusion assay and Chequerboard method.Results As for disk diffusion method, ibuprofen produced a zone of growth inhibition with diameters of (20.1±3.9) mm at 48 hours of incubation. As for broth microdilution method, the minimal inhibitory concentration (MIC) ranges of ibuprofen against aspergillus spp. were 1000-2000 μg/ml, and the minimal fungicidal concentration (MFC) ranges of that was 2000-8000 μg/ml. For 2 of 5 isolates, when ibuprofen combined with itraconazole or voriconazole, the zones of growth inhibition were larger than those of the individual drug. The results of Chequerboard method showed that fractional inhibitory concentration index (FICI) ranges were 1.125-2.500.Conclusions Ibuprofen is active against aspergillus spp.. And ibuprofen does not affect the in vitro activity of itraconazole, voriconazole, amphotericin B or micafungin against aspergillus spp..

  12. Antifungal Activity of Bee Venom and Sweet Bee Venom against Clinically Isolated Candida albicans

    Directory of Open Access Journals (Sweden)

    Seung-Bae Lee

    2016-03-01

    Full Text Available Objectives: The purpose of this study was to investigate the antifungal effect of bee venom (BV and sweet bee venom (SBV against Candida albicans (C. albicans clinical isolates. Methods: In this study, BV and SBV were examined for antifungal activities against the Korean Collection for Type Cultures (KCTC strain and 10 clinical isolates of C. albicans. The disk diffusion method was used to measure the antifungal activity and minimum inhibitory concentration (MIC assays were performed by using a broth microdilution method. Also, a killing curve assay was conducted to investigate the kinetics of the anti- fungal action. Results: BV and SBV showed antifungal activity against 10 clinical isolates of C. albicans that were cultured from blood and the vagina by using disk diffusion method. The MIC values obtained for clinical isolates by using the broth microdilution method varied from 62.5 μg/ mL to 125 μg/mL for BV and from 15.63 μg/mL to 62.5 μg/mL for SBV. In the killing-curve assay, SBV behaved as amphotericin B, which was used as positive control, did. The antifungal efficacy of SBV was much higher than that of BV. Conclusion: BV and SBV showed antifungal activity against C. albicans clinical strains that were isolated from blood and the vagina. Especially, SBV might be a candidate for a new antifungal agent against C. albicans clinical isolates.

  13. Comparison of methods for total genomic DNA extraction from Sichuan pickled vegetables broth%四川泡菜盐卤中微生物总基因组DNA提取方法的比较

    Institute of Scientific and Technical Information of China (English)

    蒋云露; 杨建涛; 王猛; 罗军; 蒋倩; 姜露熙; 张琴; 饶瑜; 马力

    2015-01-01

    了得到较高质量的基因组脱氧核糖核酸(DNA),本实验采取了5种方法对四川泡菜盐卤总基因组进行提取,对其提取的浓度、纯度进行分析比较,并以此为模版进行聚合酶链反应(PCR)体外扩增.结果表明,F1所得的基因组DNA纯度及浓度最高,F4方法提取的基因组DNA浓度较高,但受到蛋白或酚类物质的污染,F2以及F5两种方法得到的基因组DNA浓度较低,F3所得的基因组DNA浓度低并且含有较多RNA杂质.因此,本研究采用的5种对四川泡菜盐卤进行总基因组提取方法中,方法F1最合理、高效.

  14. Susceptibility of anaerobic bacteria to metronidazole, ornidazole, and tinidazole and routine susceptibility testing by standardized methods.

    Science.gov (United States)

    Wust, J

    1977-04-01

    A total of 114 strains of anaerobic bacteria were examined for their susceptibility to metronidazole, ornidazole, and tinidazole by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration in different media. All strains, with the exception of the isolates of Propionibacterium acnes, were inhibited by 3.1 mug each and killed by 6.3 mug each of all three nitroimidazole compounds per ml. No significant differences in MIC values were found among metronidazole, ornidazole, and tinidazole. Only minor differences were detected by comparing MIC values obtained in brain heart infusion agar with and without sheep blood, brucella agar, and Mueller-Hinton agar (both containing blood). When the strains were tested by the modified broth-disk method proposed by the Anaerobe Laboratory of the Virginia Polytechnic Institute (VPI), there was good correlation with the MIC values (97.4% agreement for metronidazole and 94.7% for ornidazole and tinidazole). For routine testing, use of a 30-mug-class disk of either nitroimidazole derivative is proposed for the broth-disk method, resulting in a final concentration of 6 mug/ml in the test tubes, a concentration easily attainable in body fluids. In contrast to the broth-disk method, there was very poor correlation between inhibition zone diameters by the standardized VPI agar diffusion test and MIC values. PMID:856015

  15. Too many cooks spoil the broth?

    DEFF Research Database (Denmark)

    Rørbæk, Anne

    To collaborate across organisational contexts is not an easy task, but to do so and to succeed can be vital for reaching creative and innovative results. This paper is an empirical exploration of contextual challenges for collaborative museum design, focusing on the interplay between museums and ...... ICT design companies. Based on empirical findings, two kinds of challenges are presented as particularly interesting in relation to this context: Structure related challenges, and value and practice related challenges....

  16. Thermal tolerance characteristics of non-O157 Shiga toxigenic strains of Escherichia coli (STEC) in a beef broth model system are similar to those of O157:H7 STEC.

    Science.gov (United States)

    Vasan, Akhila; Leong, Wan Mei; Ingham, Steve C; Ingham, Barbara H

    2013-07-01

    The non-O157 Shiga toxigenic Escherichia coli (STEC) serogroups most commonly associated with illness are O26, O45, O103, O111, O121, and O145. In the United States, these serogroups are considered adulterants in raw nonintact beef. To begin to understand the behavior of these pathogens in meat systems, we compared the thermal tolerance of acid-adapted cells of non-O157 STEC and O157:H7 STEC in a beef-derived broth. D58°C-values were determined for at least three strains per serogroup, and D54.6°C-values and D63.6°C-values were determined for one strain per serogroup. Each strain was grown to stationary phase in brain heart infusion broth (BHIB; pH 7.0) and inoculated into prewarmed BHIB in a shaking water bath for thermotolerance experiments at 54.6, 58.0, or 63.6°C (three trials per strain). Samples were heated for up to 160 min at 54.6°C, 3 min at 58.0°C, or 45 s at 63.6°C, with periodic sampling followed by rapid cooling and plating on modified Levine's eosin methylene blue agar. For each strain and temperature, the log CFU per milliliter was plotted versus time, and D-values were determined. Across all strains, the least and most heat tolerant STEC serogroups at 58°C were O145 and O157, respectively. D58°C-values in BHIB ranged from 0.44 min for an O145 strain to 1.42 min for an O157:H7 strain. D58°C-values for O157 STEC strains were significantly higher than those for at least one strain in each of the non-O157 STEC serogroups (P < 0.05) except for serogroup O103. At 54.6°C, the most heat-resistant STEC strain belonged to serogroup O103 and was significantly more heat tolerant than the O157:H7 strains (P < 0.05). Grouping the strains, there were no significant differences in heat tolerance between O157 and non-O157 STEC at 63.6°C (P ≥ 0.05). The z-values for non-O157 STEC strains were comparable to those for O157:H7 STEC strains (P ≥ 0.05), ranging from 4.10 to 5.21°C. These results suggest that thermal processing interventions that target

  17. 基于 AOPAN 纳米纤维膜的粘杆菌素发酵液后处理研究%Study on preparation of AOPAN nanofibrous membrane and their applications for filtration treatment of colistin fermentation broth

    Institute of Scientific and Technical Information of China (English)

    凤权; 魏安静; 侯大寅; 武丁胜

    2014-01-01

    Polyacryonitrile ( PAN) nanofibers were prepared by electrospinning, and amidoximepolyacryonitrile ( AOPAN) nanofibrous membranes were generated by chemical modification of PAN nanofibers, which were used as the filtration membrane for separation of colistin fermentation broth.According to the experimental data, characters materials and comparing nanofibrous membranes filtration separation performance, double-layer nanofibrousmembrane was selected.The optimum operating parameters had been determined:pressure was 0.14 Mpa.Under these optimum parameters, the flux of the nanofibrous membrane was 2.61 L/m2 .min, the protein reta-tion ratio was is nearly 90%, and the part of pigment and other impurities had been removed.%使用静电纺丝再经胺肟化改性制备AOPAN纳米纤维膜,并基于AOPAN纳米纤维膜构建膜分离装置,用于粘杆菌素发酵液的后处理。研究中,对不同厚度的纳米纤维膜的渗透通量及分离性能进行比较,最终确定双层叠合的纳米纤维膜为分离膜,最适操作压力为0.14 MPa。在此条件下,分离膜的渗透通量为2.61 L/m2.min,蛋白质的截留率达到90%,色素等其他杂质也得到有效去除。

  18. 寡雄腐霉发酵液对烤烟的促生及防黑胫病效应分析%Effect of Pythium oligandrum broth on growth promotion and black shank control of flue-cured tobacco

    Institute of Scientific and Technical Information of China (English)

    贾瑞莲; 耿明明; 李艳; 段玉琪; 杜如万; 吴叶宽; 黄建国

    2016-01-01

    为了研制高效、无毒、成本低廉的生物农药,安全有效地防治烟草黑胫病,利用自主分离的寡雄腐霉生防菌株(Pythium oligandrum CQ2010)制备发酵液(Pythium oligandrum broth,POB),通过拮抗、盆栽和田间试验研究了POB对烤烟生长和黑胫病的防治效果,以及对小鼠的急性毒性.拮抗试验表明,POB能显著抑制离体烟草黑胫病菌的菌丝生长和孢子萌发.在盆栽试验中,POB能显著提高烟草叶片叶绿素含量(质量分数),增强烟株根系活力,增加烤烟对氮、磷、钾的吸收,促进烟苗生长,生物量比对照提高58.91%,效果优于寡雄腐霉卵孢子制剂.POB还能激活烟草叶片中与抗病性相关酶如过氧化物酶和过氧化氢酶的活性,诱导烟株产生抗病性反应,有利于消除活性氧和过氧化氢,使病害减轻.烤烟幼苗接种黑胫病菌后,施用POB使烟株发病率下降42.95%~65.77%,相对防治效果达到46.99%~57.32%,并显著降低烟草叶片中丙二醛含量,减轻病原菌对细胞膜的伤害.在田间试验中,POB对烟草黑胫病的防治效果达47.91%,烟叶产量提高44.47%,产值增加36.33%.小鼠急性毒性试验表明,用POB大剂量灌胃给药对小鼠体质量无显著影响,供试小鼠外观和行为均无异常,心、肝、肾、肺等器官未见病理改变.因此,POB能有效防治烟草黑胫病,促进烟株生长,提高烟叶产量和产值,且对动物安全无毒.%In order to develop efficient, nontoxic and cost-effective bio-control agents against tobacco black shank, antagonism, pot and field experiments were carried out to study the effects of fermentation broth produced from a self-isolated Pythium oligandrum (strain CQ2010) on the growth and control of black shank for flue-cured tobacco, and its acute toxicity to mice. The results of antagonism experiment indicated that Pythium oligandrum broth (POB) significantly inhibited mycelial growth and spore germination of in vitro Phytophora nicotiana

  19. Isolation, Identification and Biogenic Amines-producing Ability of Lactic Acid Bacteria from The Fermentation Broth of Chinese Rice Wine%黄酒发酵液中产生物胺乳酸菌的分离鉴定与评价

    Institute of Scientific and Technical Information of China (English)

    周韩玲; 杜丽平; 孟镇; 钟其顶; 熊正河; 邹慧君; 肖冬光

    2011-01-01

    从黄酒发酵液中分离得到6株乳酸菌,16S rDNA序列分析和生理生化鉴定结果表明:菌株R1、R4、R5、R8、R20为Lactobacillus rossiae,菌株R2为Lactobacillus casei。采用平板检测法对6株乳酸菌产生物胺能力进行了评价,结果显示只有乳酸菌R1具有产生物胺的能力。利用HPLC检测对平板检测结果进行了验证,结果表明乳酸菌R1能产598.61mg/L的腐胺,其他菌株不具有产生物胺的能力。平板检测与HPLC检测结果相一致,表明平板检测可作为一种有效、操作简单、费用低的定性评价乳酸菌产生物胺能力的手段。%Six strains of lactic acid bacteria were obtained from the fermentation broth of Chinese rice wine. By using methods of physiological-biochemical and 16S rDNA tests, strains R1, R4, R5, R8 and R20 were identified as LactobaciUus rossiae, and strain R2 was identified as LactobaciUus casei. The ability of producing biogenic amines by these strains was evaluated by MRS plates and showed that only strain R1 could produce biogenic amines. HPLC confirmed that strain R1 could produce 598.61mg/L putrescine,and the others could not produce biogenic amines. The consistency of the results indicated that MRS plates analysis could be used as an effective, simple and economical method to evaluate the lactic acid bacteria's ability of producing biogenic amines.

  20. Isolation and Purification of Antifungal Components from Trichoderma harzianum Ferment Broth by High-Speed Counter-Current Chromatography%高速逆流色谱法从哈茨木霉发酵液中分离纯化抑菌活性成分

    Institute of Scientific and Technical Information of China (English)

    沙莎; 刘雷; 潘顺; 王为民

    2013-01-01

      本文采用高速逆流色谱法(HSCCC),以正己烷:乙酸乙酯:甲醇:水=7:3:5:5(v/v/v/v)作为两相溶剂体系,直接从哈茨木霉菌 Trichoderma harzianum 发酵液的乙酸乙酯提取液中分离出对立枯丝核菌Rhizoctonia solani有抑制作用的主要活性成分,经FTICR质谱及核磁共振鉴定为木霉菌素trichodermin(每324.3 mg的乙酸乙酯提取物中分离得到4 mg),对水稻纹枯病菌Thanatephorus cucmeris菌丝生长毒力EC50及EC90分别为0.69μg·mL−1、3.02μg·mL−1,此方法可用于扩大制备木霉菌素trichodermin。此外高速逆流色谱法还得到了另一对立枯丝核菌Rhizoctonia solani有明显抑制效果的活性混合组分。%We developed a high-speed counter-current chromatography (HSCCC) method and identified the major antifungal components isolated from Trichoderma harzianum broth after extraction with ethyl acetate. The HSCCC conditions were optimized and a two-phase solvent system containing n-hexane:ethyl acetate:methanol:water=7:3:5:5 (v/v/v/v) was selected for the investigation. One pure and active compound was elucidated as trichodermin using NMR and FTICR mass spectroscopy. Trichodermin exerted a potent activity in vitro against mycelial growth of Thanatephorus cucmeris with EC50=0.69 μg·mL−1 and EC90=3.02μg·mL−1. This optimized HSCCC method can be scaled up to produce a larger amount of purified trichodermin.

  1. Studies on bactericidal efficacy of pumpkin (Cucurbita moschata Duchesne) peel

    Institute of Scientific and Technical Information of China (English)

    El Zawane Kamarudin; Qamar Uddin Ahmed; Zuvairea Nazren Mohd Sirajudin; Ahmad Jalal Khan Chowdhury

    2014-01-01

    Objective: To explore the in vitro antibacterial potential of the peel of Cucurbita moschata Duchesne (tropical pumpkin) (C. moschata) against human pathogenic bacteria. Methods:In the present study, dichloromethane (DCM), methanol (MEOH) and aqueous extracts of C. moschata peel were examined for in vitro antibacterial potency against eight bacterial strains i.e. Bacillus cereus, Burkholderia cepacia, Escherichia coli, Enterococcus faecalis, Staphyloccocus aureus, Pseudomonas aerugenosa, Vibrio alginolyticus, Vibrio parahaemolyticus using Kirby-Bauer disk diffusion susceptibility and broth micro-dilution methods. Results: DCM extract of pumpkin peel exhibited the maximum zone of inhibition against Staphyloccocus aureus (21 mm) whereas aqueous extract of pumpkin peel revealed the least zone of inhibition against Escherichia coli (8 mm). MEOH extract gave maximum zone of inhibition against Pseudomonas aerugenosa (19 mm). Broth micro-dilution method showed minimum inhibitory concentration for the DCM extract against Burkholderia cepacia at 6.25 mg/mL. The minimum bactericidal concentrations were also determined to know the nature of all extracts. DCM and MEOH extracts exhibited bactericidal nature to all bacterial strains except for the Vibrio alginolyticus. The minimum bactericidal concentrations values exhibited bactericidal nature ranging from 3.12 mg/mL to 100.00 mg/mL. The screening of antimicrobial properties of different extracts of C. moschata peel revealed that the DCM extract possessed good antimicrobial efficacy compared to MEOH and aqueous extracts. Conclusions: Peel of C. moschata possesses antibacterial compounds and could be potential source for a new class of antibiotics.

  2. In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae against oral pathogens - doi: 10.4025/actascibiolsci.v34i4.8959

    Directory of Open Access Journals (Sweden)

    Francisco Augusto Burkert Del Pino

    2012-09-01

    Full Text Available In vitro tests could be a valuable tool for the evaluation of medicinal plants’ antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods were used to evaluate antimicrobial activity. Although hydroalcoholic extracts of M. arvensis did not show any antibacterial effect, its antifungal activity against C. albicans was revealed. According to the micro-dilution broth assay, MIC of the hydroalcoholic extract from leaves of M. arvensis on Candida albicans strains ranged between 625 and 2500 mg mL-1. Results suggest that M. arvensis hydroalcoholic extract may be considered a potentially antifungal agent against C. albicans, and a possible item for human antibiotic therapy.  However, further biological tests on the plant’s efficacy and side-effects are necessary before its use on humans.  

  3. Comparação de Meios de Enriquecimento e de Plaqueamento Utilizados na Pesquisa de Salmonella em Carcaças de Frango e Fezes de Aves Comparison of Different Enrichment Broth and Plating Media Used to Isolating Salmonella from Chicken Carcasses and Poultry Faeces Samples

    Directory of Open Access Journals (Sweden)

    MS Nascimento

    2000-04-01

    Full Text Available Este trabalho foi desenvolvido para avaliar, comparativamente, os caldos de enriquecimento Rapapport-novobiocina (RVN, selenito-cistinanovobiocina (SCN e tetrationato-novobiocina (TN e os meios para plaqueamento ágar Hektoen (HE, ágar MacConkey (MC, ágar Salmonella-Shigella (SS, ágar verde brilhante (VB e ágar xilose lisina desoxicolato (XLD, utilizados no isolamento de Salmonella em carcaças de frango e fezes de aves. O procedimento bacteriológico consistiu das etapas de pré-enriquecimento, enriquecimento em caldos seletivos, plaqueamento, testes bioquímicos presuntivos e confirmação sorológica com soros polivalentes anti antígenos somáticos e flagelares de Salmonella.. As fezes foram experimentalmente contaminadas com 8 sorotipos de Salmonella (Agona, Anatum, Enteritidis, Havana, Infantis, Owakam, Schwazengrund e Typhimurium e a concentração final foi aproximadamente de 1,2 x 10² ufc/g. As fezes foram inoculadas nos caldos enriquecedores e a partir daí, seguiu-se o mesmo procedimento utilizado para as carcaças. Os resultados referentes às carcaças de frango não foram estatisticamente diferentes (p>0,01 para os caldos e as placas. Todavia, verificou-se superioridade numérica em relação aos caldos SCN e TN sobre o caldo RVN, e em relação ao ágar XLD sobre os demais. Verificou-se também que com o emprego de dois caldos de enriquecimento e de dois meios para plaqueamento pode-se obter maior positividade. Quanto ao exame das fezes, o caldo TN mostrou-se superior aos demais (p>0,01, não havendo diferença (p>0,01 de resultados para os meios de plaqueamento. Os resultados sugerem que a utilização de mais de um meio de enriquecimento e de plaqueamento poderia aumentar as chances de isolamento de Salmonella.This study was undertaken to compare selenite-cystine-novobiocin (SCN broth, tetrationate-novobiocin (TN broth and Rappaport-Vassiliadisnovobiocin (RVN broth as enrichment and MacConkey (MC agar, brilliant green (BG

  4. A rapid method for the determination of microbial susceptibility using the firefly luciferase assay for adenosine triphosphate (ATP)

    Science.gov (United States)

    Vellend, H.; Tuttle, S. A.; Barza, M.; Weinstein, L.; Picciolo, G. L.; Chappelle, E. W.

    1975-01-01

    Luciferase assay for adenosine triphosphate (ATP) was optimized for pure bacteria in broth in order to evaluate if changes in bacterial ATP content could be used as a rapid measure of antibiotic effect on microorganisms. Broth cultures of log phase bacteria were incubated at 310 K (37 C) for 2.5 hours at antimicrobial concentrations which resulted in the best discrimination between sensitive and resistant strains. Eighty-seven strains of 11 bacterial species were studied for their susceptibility to 12 commonly used antimicrobial agents: ampicillin, Penicillin G, nafcillin, carbenicillin, cephalothin, tetracycline, erythromycin, clindamycin, gentamicin, nitrofurantoin, colistin, and chloramplenicol. The major advantage of the ATP system over existing methods of rapid microbial susceptibility testing is that the assay can be made specific for bacterial ATP.

  5. Er CAPM brothætt eða kannski and-brothætt?

    OpenAIRE

    Ársæll Valfells 1972

    2013-01-01

    Meira en fimmtíu ár eru frá liðin því kenningar um aðferðir til verðlagningar fjármálaeigna komu fram hjá Treynor, Sharpe, Lintner og Mossin og notaðar eru til samhliða kenningum Sharpe, Markowitz og Miller við stýringu eignasafna. Flestar kennslubækur sem og bolur fjármálafræði hefur stuðst við framan- greindar kenningar, þó með nokkuð óbragð í munni, þar sem þeir sem reyna að beita þessum kenningum uppgötva fljótt að þær virðast betri á blaði en borði. Á síðustu 20 árum he...

  6. A comparison of standard cultural methods for the detection of foodborne Salmonella species including three new chromogenic plating media.

    Science.gov (United States)

    Schönenbrücher, Vanessa; Mallinson, Edward T; Bülte, Michael

    2008-03-31

    In this study the draft of the horizontal method for the detection of Salmonella species from human food and animal feed (ISO 6579:2002) was compared to the European gold standard (DIN EN 12824:1998), including the three new chromogenic plating media AES Salmonella Agar Plate (ASAP), Oxoid Salmonella Chromogen Media (OSCM) and Miller-Mallinson agar (MM). First the growth and appearance of 36 bacterial type strains (Salmonella and other 21 species) on ASAP, OSCM and MM were compared to those on the three traditional agars Brilliant Green Agar according to Edel and Kampelmacher (BGA), Xylose Lysine Deoxycholate Agar (XLD) and Xylose Lysine Tergitol 4 Agar (XLT4). Only on MM agar, did all of 36 tested type strains produce typical colonies, especially strains of S. Senftenberg, Salmonella arizonae, S. Dublin and S. Derby. Artificial inoculation experiments using raw pork ground meat (n=92) were subsequently conducted. A shortened incubation time of 24 h in RVS broth yielded a Salmonella species recovery of 100% from spiked meat samples. Finally, 286 naturally contaminated raw porcine and bovine minced meat samples and raw poultry meat samples were investigated. Forty-three strains from a total of 39 Salmonella-positive samples were found. S. Typhimurium (n=21), with DT 104 L, DT 012 and RDNC being the most prevalent subtypes isolated. D-tartrate-positive S. Paratyphi B (n=2) and S. Saint-Paul (n=3) were also recovered. They were cultured from poultry meat and were multi-resistant against antibiotics including nalidixic acid. Rappaport Vassiliadis broth with soypeptone (RVS) yielded the highest recovery of Salmonella spp. (97,4%) compared to Tetrathionate broth with Novobiocin according to Muller and Kauffman (MKTTn, 94,9%) and Selenite Cystine broth (SC, 38,5%). However, no significant difference was obtained by comparing the ISO 6579:2002 draft to the gold standard. PMID:18192050

  7. A rapid and gentle method for isolation of genomic DNA from pathogenic Nocardia spp.

    OpenAIRE

    Torres, R D; Oletta, C A; Zlotnik, H

    1996-01-01

    The lack of simple and efficient methods for extraction of DNA from Nocardia spp. has hampered molecular manipulation of the DNA for diagnostic purposes. In the present study, a method for the rapid extraction of undegraded genomic nocardial DNA was established. Briefly, 14 pathogenic Nocardia strains were grown at 37 degrees C for 3 to 5 days in Sauton broth containing 0.05% Tween 80. Subsequently, the cultures were treated for 48 h with 1.2 mg of cycloserine per ml (final concentration). Ce...

  8. Separation of Crocin and Picrocrocin from Saffron Cell Culture Broth by Macroporous Resin Adsorption%利用大孔吸附树脂吸附分离藏红花细胞培养液中藏红花素和藏红花苦素

    Institute of Scientific and Technical Information of China (English)

    吴频梅; 袁丽红; 黄晶; 孙镇

    2011-01-01

    Objective: The separation of crocin and picrocrocin from saffron cell culture broth was investigated by macroporous resin adsorption.Methods: The effectiveness of 4 types of macroporous resin was compared in separating crocin and picrocrocin.The application of HPD-100A resin for separating crocin and picrocrocin from saffron cell culture broth was optimized.Results: HPD-100A resin was the best resin to separate crocin and picrocrocin from saffron cell culture broth among the investigated ones.The best separation of crocin and picrocrocin was achieved as an adsorption rate of 94.4% and 75.5% and a desorption rate of 99.9% and 87.5%,respectively when the cell culture broth was concentrated until a concentration of 1.0 mg/mL and 24.6 mg/mL for crocin and picrocrocin,respectively,temperature 25 ℃,pH 6,and 1.5 BV of each concentrate was loaded onto the column at a flow rate of 1.5 mL/min after being adjusted to pH 6 and then eluted with 1.7 BV of 40% aqueous ethanol at a flow rate of 1.0 mL/min.Conclusion: Macroporous resin adsorption is feasible and holds promise for separating crocin and picrocrocin from saffron cell culture broth.%目的:研究大孔树脂提取藏红花细胞培养液中藏红花素和藏红花苦素的工艺。方法:对4种大孔树脂提取藏红花素和藏红花苦素的效果进行比较,考察HPD-100A大孔树脂提取藏红花素和藏红花苦素的最佳工艺条件。结果:HPD-100A树脂提取藏红花素和藏红花苦素效果最佳,其最适工艺条件为25℃、色素液pH6、藏红花素和藏红花苦素上样质量浓度分别为1.0mg/mL和24.6mg/mL、溶液处理量1.5BV、吸附流速1.5mL/min、洗脱剂为体积分数40%乙醇溶液、洗脱剂体积1.7BV、洗脱流速1.0mL/min。藏红花素和藏红花苦素的吸附率分别达到94.4%和75.5%,解吸率分别为99.9%和87.5%。结论:采用大孔吸附树脂吸附分离藏红花培养液中藏红花素和藏红花苦素方法可行,前景广阔。

  9. Travel-Related Carbapenemase-Producing Gram-Negative Bacteria in Alberta, Canada: the First 3 Years

    OpenAIRE

    Peirano, Gisele; Ahmed-Bentley, Jasmine; Fuller, Jeff; Rubin, Joseph E; Pitout, Johann D. D.

    2014-01-01

    We describe here the characteristics of Alberta, Canada, patients with infections or colonizations with carbapenemase-producing Gram-negative bacteria during 2010 to 2013 that were linked to recent travel outside Canada. Antimicrobial susceptibility was determined by broth microdilution, and isolates were characterized using PCR, sequencing, and multilocus sequencing typing. A broth mating study was used to assess the transferability of resistance plasmids, which were subsequently characteriz...

  10. Neem (Azadirachta indica A. Juss Oil: A Natural Preservative to Control Meat Spoilage

    Directory of Open Access Journals (Sweden)

    Paola Del Serrone

    2015-01-01

    Full Text Available Plant-derived extracts (PDEs are a source of biologically-active substances having antimicrobial properties. The aim of this study was to evaluate the potential of neem oil (NO as a preservative of fresh retail meat. The antibacterial activity of NO against Carnobacterium maltaromaticum, Brochothrix thermosphacta, Escherichia coli, Pseudomonas fluorescens, Lactobacillus curvatus and L. sakei was assessed in a broth model system. The bacterial growth inhibition zone (mm ranged from 18.83 ± 1.18 to 30.00 ± 1.00, as was found by a disc diffusion test with 100 µL NO. The bacterial percent growth reduction ranged from 30.81 ± 2.08 to 99.70 ± 1.53 in the broth microdilution method at different NO concentrations (1:10 to 1:100,000. Viable bacterial cells were detected in experimentally-contaminated meat up to the second day after NO treatment (100 µL NO per 10 g meat, except for C. maltaromaticum, which was detected up to the sixth day by PCR and nested PCR with propidium monoazide (PMA™ dye. In comparison to the previously published results, C. maltaromaticum, E. coli, L. curvatus and L. sakei appeared more susceptible to NO compared to neem cake extract (NCE by using a broth model system.

  11. Presence of an Active Efflux System in the Fluoroquinolones Resistance of Mycoplasma Hominis

    Institute of Scientific and Technical Information of China (English)

    姚艳冰; 吴移谋; 朱翠明; 曾铁兵; 曾焱华

    2003-01-01

    Objective: To investigate the possible presence of an active efflux system in resistance to fluoroqninolones in Mycoplasma hominis. Methods: The resistant strains of M. hominis were selected from one hundred and three clinical strains of M. homlnls by broth microdilution method. The ac-cumulation of ciprofloxacin in M. hominis and the in-fluence of carbonyl cyanide m-chlorophenyl- hydrazone (CCCP) and reserpine were measured by a fluores-cence method. Results: Two resistant strains and two susceptible strains of M. hominis were selected in vitro. The accu-mulation of ciprofloxacin for resistant strains is lower than that of susceptible strains. CCCP and reserpine had different influence on clinical strains of M.hominis. Reserpine could dramatically increase the accumulation of ciprofloxacin, however CCCP had a little effect on it. Conclusion: These results suggest that the pres-ence of an active efflux system implicated in the fluoroouinolones-resistant in M. hominis.

  12. Antioxidant and antimicrobial activities of aporphinoids and other alkaloids from the bark of Annona salzmannii A. DC. (Annonaceae).

    Science.gov (United States)

    Costa, Emmanoel Vilaça; da Cruz, Pedro Ernesto Oliveira; de Lourenço, Caroline Caramano; de Souza Moraes, Valéria Regina; de Lima Nogueira, Paulo Cesar; Salvador, Marcos José

    2013-01-01

    The antioxidant capacity by oxygen radical absorbance capacity (ORAC)-FL method and antimicrobial activity using the broth microdilution method of aporphinoids (liriodenine 1, anonaine 2 and asimilobine 3) and other alkaloids (reticuline 4 and cleistopholine 5) isolated from the bark of Annona salzmannii A. DC. (Annonaceae) were evaluated. For antioxidant activity, the most active alkaloid was asimilobine with ORAC value of 2.09 relative trolox equivalents. For antimicrobial activity, some alkaloids showed significant minimal inhibitory concentration (MIC) values in the range of 25-100 µg mL(-1). The most active compounds were the aporphinoids liriodenine, anonaine and asimilobine, some of them more active than the positive control. PMID:22582985

  13. Ericoside, a new antibacterial biflavonoid from Erica mannii (Ericaceae).

    Science.gov (United States)

    Bitchagno, Gabin Thierry M; Tankeo, Simplice B; Tsopmo, Apollinaire; Simo Mpetga, James D; Tchinda, Alembert T; Fobofou, Serge Alain T; Nkuete, Antoine Honoré L; Wessjohann, Ludger A; Kuete, Victor; Tane, Pierre

    2016-03-01

    A new dihydroflavonol-flavonol biflavonoid derivative, named ericoside was isolated from the ethanol extract of the whole plant of Erica mannii along with the known flavonoid, taxifolin 3-O-α-L-rhamnopyranoside; and two readily available sterols (sitosterol, sitosterol 3-O-β-D-glucopyranoside). The isolation was performed using chromatographic methods and the structure of purified molecules were elucidated using spectroscopic techniques (e.g. MS, NMR) and by comparison with literature data. The crude ethanol extract, ericoside, and taxifolin 3-O-α-L-rhamnopyranoside were tested against ten Gram-negative bacteria including multidrug resistant clinical isolates using a broth microdilution method. The crude ethanol extract showed no noteworthy activity. Of the purified compounds, ericoside displayed moderate activity against the resistant Escherichia coli AG100 with a MIC of 64 μg/mL. PMID:26802608

  14. An efficient synthesis of 3'-indolyl substituted pyrido[1,2-]benzimidazoles as potential antimicrobial and antioxidant agents

    Indian Academy of Sciences (India)

    Harshad G Kathrotiya; Manish P Patel

    2013-09-01

    A new class of indole-based pyrido[1,2-]benzimidazole derivatives 4a-r have been synthesized by one-pot cyclocondensation reaction of 2-phenyl-1-indole-3-carboxaldehyde 1a-i, malononitrile 2 and 2-cyanomethylbenzimidazole 3a-b in the presence of catalytic amount of NaOH. In vitro antimicrobial activity of the synthesized compounds were investigated against a representative panel of pathogenic strains specifically three Gram-positive bacteria (Streptococcus pneumoniae, Clostridium tetani, Bacillus subtilis), three Gram-negative bacteria (Salmonella typhi, Vibrio cholerae, Escherichia coli) and two fungi (Aspergillus fumigatus, Candida albicans) using broth microdilution MIC (minimum inhibitory concentration) method. In vitro antioxidant activity was evaluated by ferric-reducing antioxidant power (FRAP) assay method. Compounds 4c, 4e, 4l and 4q have been found to be most efficient antimicrobial members while compounds 4h and 4p possess better ferric reducing antioxidant power.

  15. Direct and indirect antimicrobial effects of α-mangostin on pathogenic microorganisms

    Institute of Scientific and Technical Information of China (English)

    Nurul Huda Syed Ibrahim; Muhammad Taher; Deny Susanti; Qamar Uddin Ahmed

    2014-01-01

    Objective: To test direct and indirect antimicrobial properties of α-mangostin towards a number of bacteria and fungi.Methods:Activity of α-mangostin paired with an antibiotic was studied by calculating its fractional inhibitory concentration (FIC).Results:The experiment was carried out using broth microdilution and checkerboards methods. tetracycline showed no interaction with the combination with α-mangostin where the FIC indexes were between the range of 0.54. Activity of doxycycline on Pseudomonas aeruginosa fell into other set of range, FICindex≥4 which is an antagonism. The activity of all four bacteria towards ampicillin, penicillin G, streptomycin and Conclusions: The FIC index is far away in the range. The coupled antibiotic and α-mangostin is considered synergy in action if it lies in FICindex≤0.5 and it was found that the isolated compound,α-mangostin revealed very low synergistic antimicrobial effects when coupled with antibiotics.

  16. Evaluation of Traditional Chinese Medicinal Plants for Anti-MRSA Activity with Reference to the Treatment Record of Infectious Diseases

    Directory of Open Access Journals (Sweden)

    Zhong-Qi Bian

    2012-03-01

    Full Text Available The in vitro antimicrobial activities of 30 Chinese medicinal plants were evaluated with reference to the treatment record of infectious diseases in the Traditional Chinese Medicine (TCM literature. The plant materials were extracted with 80% ethanol and the extracts were primarily screened against conventional clinical pathogens like Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Candida albicans by the agar diffusion method. Their inhibition zone diameters (IZDs, mm, 50 mg/mL ranged from 2,048 by the standard broth microdilution method. The seven extracts from M. yunnanensis, S. sinensis, G. morella, E. daneillii, M. squamulata, S. arborescens and B. hancei were determined as the most active extracts, with MICs of 8–64 μg/mL. The results were in good agreement with their traditional applications in skin and other infections.

  17. Miniaturized Antimicrobial Susceptibility Test by Combining Concentration Gradient Generation and Rapid Cell Culturing

    Directory of Open Access Journals (Sweden)

    Samuel C. Kim

    2015-10-01

    Full Text Available Effective treatment of bacterial infection relies on timely diagnosis and proper prescription of antibiotic drugs. The antimicrobial susceptibility test (AST is one of the most crucial experimental procedures, providing the baseline information for choosing effective antibiotic agents and their dosages. Conventional methods, however, require long incubation times or significant instrumentation costs to obtain test results. We propose a lab-on-a-chip approach to perform AST in a simple, economic, and rapid manner. Our assay platform miniaturizes the standard broth microdilution method on a microfluidic device (20 × 20 mm that generates an antibiotic concentration gradient and delivers antibiotic-containing culture media to eight 30-nL chambers for cell culture. When tested with 20 μL samples of a model bacterial strain (E. coli ATCC 25922 treated with ampicillin or streptomycin, our method allows for the determination of minimum inhibitory concentrations consistent with the microdilution test in three hours, which is almost a factor of ten more rapid than the standard method.

  18. A microtiter assay shows effectivness of fungicides for control of Colletotrichum spp from strawberry

    Science.gov (United States)

    The anthracnose pathogens, Colletotrichum acutatum, C. gloeosporioides and C. fragariae, have developed resistance to some fungicides. We used a micro-dilution broth assay to test in vitro the activity of 16 chemicals against ten isolates of Colletotrichum spp. using a dose-response format. At a c...

  19. Oral antibiotics for infections due to multidrug-resistant Gram-negative organisms.

    Science.gov (United States)

    Glasser, Jessie S; Markelz, Ana E; Zera, Wendy C; Beckius, Miriam L; Mende, Katrin; Murray, Clinton K

    2011-08-01

    We determined minimum inhibitory concentrations of rifampicin, nitrofurantoin, amoxicillin-clavulanic acid, and cefdinir, plus a combination of amoxicillin-clavulanic acid and cefdinir by broth microdilution for mainly wound isolates of Escherichia coli and Klebsiella pneumoniae. E. coli and K. pneumoniae susceptibilities increased by combining amoxicillin-clavulanic acid and cefdinir. PMID:21466257

  20. Activities of five new fluoroquinolones against Pseudomonas pseudomallei.

    OpenAIRE

    Winton, M D; Everett, E D; Dolan, S A

    1988-01-01

    Thirty-four isolates of Pseudomonas pseudomallei were tested by a broth microdilution technique for susceptibility to amifloxacin, ciprofloxacin, enoxacin, norfloxacin, and ofloxacin. Ciprofloxacin was the most active agent tested, with an MIC for 90% of the strains tested of 8 micrograms/ml. These in vitro results suggest that the fluoroquinolones tested would not be optimal for therapy of melioidosis.

  1. Polyclonal Mycobacterium avium infections in patients with AIDS: variations in antimicrobial susceptibilities of different strains of M. avium isolated from the same patient.

    Science.gov (United States)

    von Reyn, C F; Jacobs, N J; Arbeit, R D; Maslow, J N; Niemczyk, S

    1995-01-01

    Broth microdilution MICs were determined for pairs of strains isolated from five AIDS patients with polyclonal Mycobacterium avium infection. Four (80%) of the five patients were infected simultaneously with strains having different antimicrobial susceptibility patterns. These findings have implications for the interpretation of susceptibility data in M. avium prophylaxis and treatment trials. PMID:7790424

  2. Polyclonal Mycobacterium avium infections in patients with AIDS: variations in antimicrobial susceptibilities of different strains of M. avium isolated from the same patient.

    OpenAIRE

    von Reyn, C F; Jacobs, N J; Arbeit, R D; Maslow, J.N.; Niemczyk, S

    1995-01-01

    Broth microdilution MICs were determined for pairs of strains isolated from five AIDS patients with polyclonal Mycobacterium avium infection. Four (80%) of the five patients were infected simultaneously with strains having different antimicrobial susceptibility patterns. These findings have implications for the interpretation of susceptibility data in M. avium prophylaxis and treatment trials.

  3. Effect of Manganese in Test Media on In Vitro Susceptibility of Enterobacteriaceae and Acinetobacter baumannii to Tigecycline

    OpenAIRE

    Veenemans, J; Mouton, J.; Kluijtmans, J.A.J.W.; Donnely, R.; Verhulst, C.; Keulen, van, H.

    2012-01-01

    We assessed the effect of increasing manganese concentrations in test media (0.001 to 1,024 mg/liter) on MICs of tigecycline. For both broth microdilution (BMD) and Etests, this effect was negligible for physiological concentrations, but MICs increased when concentrations exceeded 8 mg/liter. Susceptibility testing should be performed on media with standardized low manganese content.

  4. In vitro antibacterial screening of Cryptolepis sanguinolenta alkaloids.

    Science.gov (United States)

    Paulo, A; Duarte, A; Gomes, E T

    1994-10-01

    The ethanol and aqueous crude extracts and five alkaloids isolated from the roots of Crytolepis sanguinolenta (Lindl.) Schlechter were screened for antibacterial activity against 7 reference strains by the twofold serial broth microdilution assay. The ethanol extract and the alkaloids cryptolepine and cryptoheptine inhibited the growth of all strains tested except that of Pseudomonas aeruginosa. PMID:7853864

  5. Evaluation of antibacterial effects of Zataria multiflora Boiss extracts against ESBL-producing Klebsiella pneumoniae strains

    Directory of Open Access Journals (Sweden)

    Masoud Dadashi

    2016-04-01

    Full Text Available Objective: There are few therapeutic options for treatment of multidrug resistant Klebsiella pneumoniae isolates as a hospital infectious agent (nosocomial infection. The aim of this study was to evaluate the antibacterial activity of Zataria multiflora Boiss extracts against ESBL-producing Klebsiella pneumoniae strains. Materials and Methods: This study was conducted on 100 K. pneumoniae isolates from two hospitals in Tehran, Iran. Antibiotic susceptibility tests were performed by Kirby-Bauer disc diffusion and microdilution broth methods and detection of ESBL was carried out according to CLSI guidelines. The blaCTX-M-15plasmid genewas detected by PCR and sequencing methods. Extracts susceptibility test was performed by broth microdilution method.  Results: Among 100 K. pneumoniae strains, 48 (48% were ESBL positive. In this study, fosfomycin, colistin and tigecycline were more active than other antibiotics. The existence of blaCTX-M-15 was detected in 30 (62.5% of 48 ESBL-producing isolates. The chloroformic extract showed potent activity against ESBL-producing K. pneumoniae strains (MIC50 = 1.56 mg/ml and MIC90=3.12mg/ml. The MIC50 and MIC90 (The MIC50 represents the MIC value at which ≥50% of the isolates in a test population are inhibited and the MIC90 represents the MIC value at which ≥90% of the strains within a test population are inhibited were 3.12 and 6.25 mg/ml and 6.25  and 12.5 mg/ml for methanolic and acetonic extracts, respectively. Conclusions: The incidence of ESBL-producing K. pneumoniae is very high. Therefore, detection of ESBL-producing K. pneumoniae isolates is of great importance in identifying drug resistance patterns in K. pneumoniae isolates and in control of infections. Zataria multiflora may have the potential to be used against multidrug resistant organisms such as clinical isolates of ESBL-producing K. pneumoniae.

  6. A combined enrichment/polymerase chain reaction based method for the routine screening of Streptococcus agalactiae in pregnant women

    Directory of Open Access Journals (Sweden)

    F.M. Munari

    2012-03-01

    Full Text Available Group B Streptococcus (GBS is the most common cause of life-threatening infection in neonates. Guidelines from CDC recommend universal screening of pregnant women for rectovaginal GBS colonization. The objective of this study was to compare the performance of a combined enrichment/PCR based method targeting the atr gene in relation to culture using enrichment with selective broth medium (standard method to identify the presence of GBS in pregnant women. Rectovaginal GBS samples from women at ≥36 weeks of pregnancy were obtained with a swab and analyzed by the two methods. A total of 89 samples were evaluated. The prevalence of positive results for GBS detection was considerable higher when assessed by the combined enrichment/PCR method than with the standard method (35.9% versus 22.5%, respectively. The results demonstrated that the use of selective enrichment broth followed by PCR targeting the atr gene is a highly sensitive, specific and accurate test for GBS screening in pregnant women, allowing the detection of the bacteria even in lightly colonized patients. This PCR methodology may provide a useful diagnostic tool for GBS detection and contributes for a more accurate and effective intrapartum antibiotic and lower newborn mortality and morbidity.

  7. Evaluation of different analysis and identification methods for Salmonella detection in surface drinking water sources

    International Nuclear Information System (INIS)

    The standard method for detecting Salmonella generally analyzes food or fecal samples. Salmonella often occur in relatively low concentrations in environmental waters. Therefore, some form of concentration and proliferation may be needed. This study compares three Salmonella analysis methods and develops a new Salmonella detection procedure for use in environmental water samples. The new procedure for Salmonella detection include water concentration, nutrient broth enrichment, selection of Salmonella containing broth by PCR, isolation of Salmonella strains by selective culture plates, detection of possible Salmonella isolate by PCR, and biochemical testing. Serological assay and pulsed-field gel electrophoresis (PFGE) can be used to identify Salmonella serotype and genotype, respectively. This study analyzed 116 raw water samples taken from 18 water plants and belonging to 5 watersheds. Of these 116, 10 water samples (8.6%) taken from 7 water plants and belonging to 4 watersheds were positive for a Salmonella-specific polymerase chain reaction targeting the invA gene. Guided by serological assay results, this study identified 7 cultured Salmonella isolates as Salmonella enterica serovar: Alnaby, Enteritidis, Houten, Montevideo, Newport, Paratyphi B var. Java, and Victoria. These seven Salmonella serovars were identified in clinical cases for the same geographical areas, but only one of them was 100% homologous with clinical cases in the PFGE pattern. - Research highlights: → A new Salmonella detecting procedure for environmental water is developed. → Salmonella isolates are identified by serological assay and PFGE. → A total of seven Salmonella serovars is isolated from environmental water.

  8. Can herbal remedies be the answer to multidrug resistance? Profile of drug resistance in Salmonella species in Eastern Cape, South Africa

    Directory of Open Access Journals (Sweden)

    Sandeep Vasaikar

    2012-04-01

    Full Text Available Objective: The treatment of serious Salmonella infections which requires the use of cephalosporins and fluoroquinolones is being compromised by the emergence of extended-spectrum beta-lactamases (ESBLs. This study reports the antibiotic profile of Salmonella species, highlighting increasing ESBLs trends in Salmonella spp. and the emergence of multi-drug resistance (MDR. To proffer solution to the problem of MDR, screening of selected herbal plants was carried out. Methods: 142 consecutive isolates of Salmonella spp. collected over a period of 4 years were tested for antibiotic resistance. Antibiogram, ESBL phenotype and confirmation of isolate were determined using a semi-automated antibiotic test. Tests were performed based on Clinical Laboratory Standards Institute standards for broth microdilution methods and interpretation using Escherichia coli ATCC 25922 as the control strain. Antibiotic resistant patterns were determined, ranking order of importance as percent (% of each type of resistance. Twelve plants selected based on ethnobotanical survey information as remedy in the treatment of stomach related ailments were screened using broth microdilution methods against strains of Salmonella, Shigella, Escherichia, Staphylococcus, Pseudomonas and Enterococcus. Results: A greater proportion of isolates were obtained from invasive cultures. Of the Salmonella isolates, there was a striking predominance of S.enterica serotype Typhi followed by S.enterica serotype Typhimurium. Most species showed pentavalent resistance to commonly used drugs. Antimicrobial resistance in S.enterica serotype Typhi is visibly increasing. Of growing concern is the increase in strains exhibiting ESBLs. Plant screening revealed promising therapeutic values in Aloe arborescens, A.striatula, and Psidium guajava. Conclusion: Increasing MDR in Salmonella serovars involved ESBLs’ production. Plants with significant antibacterial activities were comparable to the tested

  9. Amphotericin B and caspofungin resistance in Candida glabrata isolates recovered from a critically ill patient

    DEFF Research Database (Denmark)

    Krogh-Madsen, Mikkel; Arendrup, Maiken Cavling; Heslet, Lars;

    2006-01-01

    also resistant to azoles and caspofungin. In this study, 4 isolates were studied thoroughly using susceptibility assays and a mouse model and to determine clonality. METHODS: Different broth microdilution tests, Etests, and time-kill studies for antifungals were performed in different media. Three of...... the 4 isolates were examined in an in vivo experiment, in which mice were challenged intravenously with 1 of 3 isolates and treated daily with amphotericin B, caspofungin, or saline. For the clonality studies, arbitrarily primed polymerase chain reaction (PCR) was performed with the 4 isolates, 8...... differentiate between amphotericin B-susceptible and -resistant isolates. All assays identified caspofungin-susceptible and -resistant isolates. Arbitrarily primed PCR showed that the 4 isolates probably were of clonal origin. CONCLUSIONS: We have documented the emergence of amphotericin B-resistant and...

  10. Synthesis and antileishmanial and antimicrobial activities of some 2,3-disubstituted 3H-quinazolin-4-ones.

    Science.gov (United States)

    Arfan, Muhammad; Khan, Rasool; Khan, Murad Ali; Anjum, Shazia; Choudhary, Muhammad Iqbal; Ahmad, Manzoor

    2010-08-01

    A series of 2,3-disubstituted 3H-quinazolin-4-ones was synthesized. Antimicrobial activities of the synthesized compounds were investigated against Gram (+ve) and Gram (-ve) bacteria, including B. subtilis, S. aureus, S. flexneri, P. aeruginosa, and S. typhi, and six fungi, namely Trichophyton longifusus, Candida albicans, Aspergillus flavus, Microsporum canis, Fusarium solani, and Candida glabrata using the broth microdilution method. Compounds 9, 11, and 12 showed significant activities against the selected bacterial cultures, while 7-10, 12, 15, and 16 showed good to moderate antifungal activities. Compound 11 exhibited strongest leishmanicidal activity against Leishmania major (MHOM/PK/88/DESTO) promastigotes, while other compounds showed weak to moderate leishmanicidal activities. PMID:20235747

  11. Synthesis and antimicrobial activities of new higher amino acid Schiff base derivatives of 6-aminopenicillanic acid and 7-aminocephalosporanic acid

    Science.gov (United States)

    Özdemir (nee Güngör), Özlem; Gürkan, Perihan; Özçelik, Berrin; Oyardı, Özlem

    2016-02-01

    Novel β-lactam derivatives (1c-3c) (1d-3d) were produced by using 6-aminopenicillanic acid (6-APA), 7-aminocephalosporanic acid (7-ACA) and the higher amino acid Schiff bases. The synthesized compounds were characterized by elemental analysis, IR, 1H/13C NMR and UV-vis spectra. Antibacterial activities of all the higher amino acid Schiff bases (1a-3a) (1b-3b) and β-lactam derivatives were screened against three gram negative bacteria (Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Acinetobacter baumannii RSKK 02026), three gram positive bacteria (Staphylococcus aureus ATCC 25923, Enterococcus faecalis ATCC 07005, Bacillus subtilis ATCC 6633) and their drug-resistant isolates by using broth microdilution method. Two fungi (Candida albicans and Candida krusei) were used for antifungal activity.

  12. Carboxymethylated chitosan-stabilized copper nanoparticles: a promise to contribute a potent antifungal and antibacterial agent

    Energy Technology Data Exchange (ETDEWEB)

    Tantubay, Sangeeta, E-mail: sang.chem2@gmail.com [Indian Institute of Technology Kharagpur, Department of Chemistry (India); Mukhopadhyay, Sourav K. [Indian Institute of Technology Kharagpur, Department of Biotechnology (India); Kalita, Himani; Konar, Suraj [Indian Institute of Technology Kharagpur, Department of Chemistry (India); Dey, Satyahari [Indian Institute of Technology Kharagpur, Department of Biotechnology (India); Pathak, Amita, E-mail: ami@chem.iitkgp.ernet.in; Pramanik, Panchanan, E-mail: ppramanik1946@yahoo.in, E-mail: pramanik1946@gmail.com [Indian Institute of Technology Kharagpur, Department of Chemistry (India)

    2015-06-15

    Carboxymethylated chitosan (CMC)-stabilized copper nanoparticles (Cu-NPs) have been synthesized via chemical reduction of copper(II)–CMC complex in aqueous medium by hydrazine under microwave irradiation in ambient atmosphere. Structural morphology, phase, and chemical compositions of CMC-stabilized Cu-NPs (CMC–Cu-NPs) have been analyzed through high-resolution transmission electron microscopy, field emission scanning electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. Antifungal and antibacterial activities of CMC–Cu-NPs have been evaluated against Candida tropicalis and Escherichia coli through agar well diffusion method, broth microdilution assay, live–dead assay, and microscopic observation. Antimicrobial activity of spherical CMC–Cu-NPs (∼4–15 nm of diameters) has been observed to be significant for both C. tropicalis and E. coli. The cytotoxicity study indicates that CMC–Cu-NPs have no significant toxic effect against normal cell line, L929.

  13. Antibacterial activity of commercially available plant-derived essential oils against oral pathogenic bacteria.

    Science.gov (United States)

    Bardají, D K R; Reis, E B; Medeiros, T C T; Lucarini, R; Crotti, A E M; Martins, C H G

    2016-05-01

    This work investigated the antibacterial activity of 15 commercially available plant-derived essential oils (EOs) against a panel of oral pathogens. The broth microdilution method afforded the minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) of the assayed EOs. The EO obtained from Cinnamomum zeylanicum (Lauraceae) (CZ-EO) displayed moderate activity against Fusobacterium nucleatum (MIC and MBC = 125 μg/mL), Actinomyces naeslundii (MIC and MBC = 125 μg/mL), Prevotella nigrescens (MIC and MBC = 125 μg/mL) and Streptococcus mutans (MIC = 200 μg/mL; MBC = 400 μg/mL). (Z)-isosafrole (85.3%) was the main chemical component of this oil. We did not detect cinnamaldehyde, previously described as the major constituent of CZ-EO, in specimens collected in other countries. PMID:26165725

  14. Synthesis of 1,2,3-Triazole Derivatives and in Vitro Antifungal Evaluation on Candida Strains

    Directory of Open Access Journals (Sweden)

    Almir G. Wanderley

    2012-05-01

    Full Text Available 1,2,3-Triazoles have been extensively studied as compounds possessing important biological activities. In this work, we describe the synthesis of ten 2-(1-aryl-1H-1,2,3-triazol-4-ylpropan-2-ols via copper catalyzed azide alkyne cycloaddition (CuAAc or click chemistry. Next the in vitro antifungal activity of these ten compounds was evaluated using the microdilution broth method against 42 isolates of four different Candida species. Among all tested compounds, the halogen substituted triazole 2-[1-(4-chlorophenyl-1H-(1,2,3triazol-4-yl]propan-2-ol, revealed the best antifungal profile, showing that further modifications could be done in the structure to obtain a better drug candidate in the future.

  15. In vitro antifungal activities of leaf extracts of Lippia alba (Verbenaceae against clinically important yeast species

    Directory of Open Access Journals (Sweden)

    Graziela Teixeira de Oliveira

    2014-04-01

    Full Text Available Introduction There are few studies reporting the antifungal activities of Lippia alba extracts. Methods A broth microdilution assay was used to evaluate the antifungal effects of Lippia alba extracts against seven yeast species of Candida and Cryptococcus. The butanol fraction was investigated by gas chromatography-mass spectrometry. Results The butanol fraction showed the highest activity against Candida glabrata. The fraction also acted synergistically with itraconazole and fluconazole against C. glabrata. The dominant compounds in the butanol fraction were 2,2,5-trimethyl-3,4-hexanedione, 3,5-dimethyl-4-octanone and hexadecane. Conclusions The butanol fraction may be a good candidate in the search for new drugs from natural products with antifungal activity.

  16. Composition and antimicrobial activity of the essential oil of the leaves of black currant (Ribes nigrum L. cultivar Čačanska crna

    Directory of Open Access Journals (Sweden)

    TODOR VULIĆ

    2010-01-01

    Full Text Available The essential oil from the leaves of the Serbian black currant cultivar Čačanska crna, obtained by hydrodistillation, was analyzed by gas chromato¬graphy-flame ionization detection and GC–mass spectrometry. The most abundant volatile compounds were D3-carene (18.7 %, b-caryophyllene (17.7 %, sabinene (11.6 %, cis-b-ocimene (10.6 % and a-terpinolene (10.6 %. The antimicrobial activity of the oil was evaluated against 14 micro-organisms, including two clinical isolated strains, using the broth microdilution method. The most susceptible micro-organisms were Escherichia coli, Streptococcus faecalis, Staphylococcus aureus, Candida albicans and Trichophyton mentagrophytes isolates. Furthermore, the flavonol aglycones in the leaves after acid hydrolysis were qualitatively and quantitatively analysed by HPLC, and quercetin was found to be the dominant compound (84 mg/g dw.

  17. Microwave assisted synthesis of novel 4h-chromene derivatives bearing phenoxypyrazole and their antimicrobial activity assess

    Directory of Open Access Journals (Sweden)

    Sangani Chetan B.

    2012-01-01

    Full Text Available A new series of 4H-chromene derivatives 4(a-p bearing 5-phenoxypyrazole nucleus has been synthesized under microwave irradiation by reaction of 5-phenoxypyrazole-4-carbaldehyde 1(a-h, malononitrile 2 and compounds (Cyclohexanedione, Dimedon 3(a-b in presence of NaOH as basic catalyst. All the compounds were screened against three Gram positive bacteria (Streptococcus pneumoniae, Clostridium tetani, Bacillus subtilis, three Gram negative bacteria (Salmonella typhi, Vibrio cholerae, Escherichia coli and two fungi (Aspergillus fumigatus, Candida albicans using broth microdilution MIC (Minimum Inhibitory Concentration method. Upon study of antimicrobial screening, it has been observed that, majority of the compounds were found to be active against Clostridium tetani and Bacillus subtilis as well as against Candida albicans as compared to standard drugs.

  18. Antifungal activity of Oleoresins used in meat industry on some toxigenic Aspergillus spp.

    Directory of Open Access Journals (Sweden)

    Šošo Vladislava M.

    2013-01-01

    Full Text Available Different spice oleoresins are widely used in meat industry. They contribute to the specific aroma and flavor of the end products, but they have also been reported to have strong antimicrobial activity. These properties open a plenty of possibilities to be used for defining the specific sensory profile of the product but also as natural food preservatives. This paper focuses on the antifungal activity of four oleoresins against different foodborne toxigenic Aspergillus species. Oleoresins used in the experiments were cayenne pepper, black pepper, garlic and rosemary oleoresins, and they were tested against following Aspergillus species: A. clavatus, A. flavus, A. fumigatus, A. niger, A. ochraceus and A. versicolor. Antifungal activity was tested using microtitre-plate-based assay incorporating resazurin as an indicator of cell growth and broth microdilution-method. [Projekat Ministarstva nauke Republike Srbije, br. III46009

  19. Rapid Detection of Polymyxin Resistance in Enterobacteriaceae.

    Science.gov (United States)

    Nordmann, Patrice; Jayol, Aurélie; Poirel, Laurent

    2016-06-01

    For identification of polymyxin resistance in Enterobacteriaceae, we developed a rapid test that detects glucose metabolization associated with bacterial growth in the presence of a defined concentration of colistin or polymyxin B. Formation of acid metabolites is evidenced by a color change (orange to yellow) of a pH indicator (red phenol). To evaluate the test, we used bacterial colonies of 135 isolates expressing various mechanisms of colistin resistance (intrinsic, chromosomally encoded, and plasmid-mediated MCR-1) and 65 colistin-susceptible isolates. Sensitivity and specificity were 99.3% and 95.4%, respectively, compared with the standard broth microdilution method. This new test is inexpensive, easy to perform, sensitive, specific, and can be completed in <2 hours. It could be useful in countries facing endemic spread of carbapenemase producers and for which polymyxins are last-resort drugs. PMID:27191712

  20. Susceptibility of bifidobacteria of animal origin to selected antimicrobial agents.

    Science.gov (United States)

    Mayrhofer, Sigrid; Mair, Christiane; Kneifel, Wolfgang; Domig, Konrad J

    2011-01-01

    Strains of the genus Bifidobacterium are frequently used as probiotics, for which the absence of acquired antimicrobial resistance has become an important safety criterion. This clarifies the need for antibiotic susceptibility data for bifidobacteria. Based on a recently published standard for antimicrobial susceptibility testing of bifidobacteria with broth microdilution method, the range of susceptibility to selected antibiotics in 117 animal bifidobacterial strains was examined. Narrow unimodal MIC distributions either situated at the low-end (chloramphenicol, linezolid, and quinupristin/dalfopristin) or high-end (kanamycin, neomycin) concentration range could be detected. In contrast, the MIC distribution of trimethoprim was multimodal. Data derived from this study can be used as a basis for reviewing or verifying present microbiological breakpoints suggested by regulatory agencies to assess the safety of these micro-organisms intended for the use in probiotics. PMID:22312561

  1. One step synthesis of pyrido[1,2-α]benzimidazole derivatives of aryloxypyrazole and their antimicrobial evaluation

    Institute of Scientific and Technical Information of China (English)

    Hardik H.Jardosh; Chetan B.Sangani; Manish P.Patel; Ranjan G.Patel

    2013-01-01

    A new series of pyrido[1,2-α]benzimidazole derivatives bearing the aryloxypyrazole nucleus have been synthesized by base-catalyzed cyclocondensation reaction through multi-component reaction (MCR) approach.All the synthesized compounds were investigated against a representative panel of pathogenic strains using broth microdilution minimum inhibitory concentration (MIC) method for their in vitro antimicrobial activity.Reviewing the data,majority of the compounds were found to be active against employed pathogens.SAR study explores that antimicrobial activity is strongly depends on the nature of the substituents at the ether linked aryl ring attached to the pyrazole unit,together with the substituent present on the C5 of the benzimidazole unit.

  2. Antimicrobial and antifungal activities of the extracts and essential oils of Bidens tripartita.

    Directory of Open Access Journals (Sweden)

    Michał Tomczyk

    2008-12-01

    Full Text Available The aim of this study was to determine the antibacterial and antifungal properties of the extracts, subextracts and essential oils of Bidens tripartita flowers and herbs. In the study, twelve extracts and two essential oils were investigated for activity against different Gram-positive Bacillus subtilis, Micrococcus luteus, Staphylococcus aureus, Gram-negative bacteria Escherichia coli, E. coli (beta-laktamase+, Klebsiella pneumoniae (ESBL+, Pseudomonas aeruginosa and some fungal organisms Candida albicans, C. parapsilosis, Aspergillus fumigatus, A. terreus using a broth microdilution and disc diffusion methods. The results obtained indicate antimicrobial activity of the tested extracts (except butanolic extracts, which however did not inhibit the growth of fungi used in this study. Bacteriostatic effect of both essential oils is insignificant, but they have strong antifungal activity. These results support the use of B. tripartita to treat a microbial infections and it is indicated as an antimicrobial and antifungal agent, which may act as pharmaceuticals and preservatives.

  3. SYNTHESIS AND BIOEVALUATION OF KETOCONAZOLE THIOSEMICARBAZONE ANALOGUES

    Directory of Open Access Journals (Sweden)

    Y. Murti

    2011-12-01

    Full Text Available Ketoconazole (KTZ is a synthetic antifungal drug used to prevent and treat fungal infections, especially in immunocompromised patients such as those with AIDS. Resistance to ketoconazole has been observed in a number of clinical fungal isolates, including C. albicans. Thus new effective agents with less toxicity against fungal infection are urgently required. With this view, ketoconazole thiosemicarbazone analogues (Compounds 1-10 were synthesized wherein condensation of different thiosemicarbazides substituted by different cyclic and aromatic amines with the KTZ was done. Investigation of in-vitro antifungal activity of compounds was done by broth microdilution assay method against five pathogenic fungi Aspergillus niger, Candida albicans, Candida krusei, Candida glabrata and Candida tropicalis. Ketoconazole was used as reference for inhibitory activity against fungi. All the compounds were found potent antifungal agents, while compounds 8, 9 and 10 exhibited excellent in-vitro antifungal activity showing importance of halogenated compounds.

  4. Differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis

    Directory of Open Access Journals (Sweden)

    Grazieli Maboni

    2015-06-01

    Full Text Available The aim of this study was to determine the differences in the antimicrobial susceptibility profiles of Moraxella bovis, M. bovoculi and M. ovis. Thirty-two strains of Moraxella spp. isolated from cattle and sheep with infectious keratoconjunctivitis were tested via broth microdilution method to determine their susceptibility to ampicillin, cefoperazone, ceftiofur, cloxacillin, enrofloxacin, florfenicol, gentamicin, neomycin, oxytetracycline and penicillin. The results demonstrated that Moraxella spp. strains could be considered sensitive for most of the antimicrobials tested in this study, but differences between the antimicrobial susceptibility profiles of these three Moraxella species were found. M. bovis might differ from other species due to the higher MIC and MBC values it presented.

  5. Antifungal susceptibility testing of Aspergillus species complex in the Clinical Laboratory: how to do it, when to do it, and how to interpret it

    Directory of Open Access Journals (Sweden)

    Esther Manso

    2014-12-01

    Full Text Available The emergence of drug resistance in fungal pathogens has a profound impact on human health given limited number of antifungal drugs. Antifungal resistance in Aspergillus spp. infection can be encountered in the antifungal drug-exposed patient due to selection of intrinsically resistant species or isolates with acquired resistance belonging to species that are normally susceptible. Resistance to triazoles is not common in Aspergillus spp., however, triazole resistance in A. fumigatus appears to be increasing in several European countries in recent years and can be clinically relevant. The Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing have developed breakpoints and epidemiological cutoff values that are now established for Aspergillus spp. Clinical microbiology laboratories will be employed commercial susceptibility assays, rather than reference broth microdilution methods and comparative studies are particularly important.

  6. Evaluation of antimicrobial activity of extracts of Tibouchina candolleana (melastomataceae, isolated compounds and semi-synthetic derivatives against endodontic bacteria

    Directory of Open Access Journals (Sweden)

    Fernanda M. dos Santos

    2012-06-01

    Full Text Available This work describes the phytochemical study of the extracts from aerial parts of Tibouchina candolleana as well as the evaluation of the antimicrobial activity of extracts, isolated compounds, and semi-synthetic derivatives of ursolic acid against endodontic bacteria. HRGC analysis of the n-hexane extract of T. candolleana allowed identification of b-amyrin, a-amyrin, and b-sitosterol as major constituents. The triterpenes ursolic acid and oleanolic acid were isolated from the methylene chloride extract and identified. In addition, the flavonoids luteolin and genistein were isolated from the ethanol extract and identified. The antimicrobial activity was investigated via determination of the minimum inhibitory concentration (MIC using the broth microdilution method. Amongst the isolated compounds, ursolic acid was the most effective against the selected endodontic bacteria. As for the semi-synthetic ursolic acid derivatives, only the methyl ester derivative potentiated the activity against Bacteroides fragilis.

  7. Antimicrobial Agents

    Directory of Open Access Journals (Sweden)

    Sigrid Mayrhofer

    2011-01-01

    Full Text Available Strains of the genus Bifidobacterium are frequently used as probiotics, for which the absence of acquired antimicrobial resistance has become an important safety criterion. This clarifies the need for antibiotic susceptibility data for bifidobacteria. Based on a recently published standard for antimicrobial susceptibility testing of bifidobacteria with broth microdilution method, the range of susceptibility to selected antibiotics in 117 animal bifidobacterial strains was examined. Narrow unimodal MIC distributions either situated at the low-end (chloramphenicol, linezolid, and quinupristin/dalfopristin or high-end (kanamycin, neomycin concentration range could be detected. In contrast, the MIC distribution of trimethoprim was multimodal. Data derived from this study can be used as a basis for reviewing or verifying present microbiological breakpoints suggested by regulatory agencies to assess the safety of these micro-organisms intended for the use in probiotics.

  8. Rapid Detection of Polymyxin Resistance in Enterobacteriaceae

    Science.gov (United States)

    Jayol, Aurélie; Poirel, Laurent

    2016-01-01

    For identification of polymyxin resistance in Enterobacteriaceae, we developed a rapid test that detects glucose metabolization associated with bacterial growth in the presence of a defined concentration of colistin or polymyxin B. Formation of acid metabolites is evidenced by a color change (orange to yellow) of a pH indicator (red phenol). To evaluate the test, we used bacterial colonies of 135 isolates expressing various mechanisms of colistin resistance (intrinsic, chromosomally encoded, and plasmid-mediated MCR-1) and 65 colistin-susceptible isolates. Sensitivity and specificity were 99.3% and 95.4%, respectively, compared with the standard broth microdilution method. This new test is inexpensive, easy to perform, sensitive, specific, and can be completed in <2 hours. It could be useful in countries facing endemic spread of carbapenemase producers and for which polymyxins are last-resort drugs. PMID:27191712

  9. Studying the antibacterial effects of some phytochemical compounds isolated from Origanum syriacum, Thymus syriacus and Myrtus communis on some bacteria

    International Nuclear Information System (INIS)

    Summary The Origanum Syriacum, Thymus syriacus and Myrtus communis leaves samples were collected. Essential oils (EO) extractions were done by hydrodistillation in a Clevenger apparatus. GC-FID was used to analyze the essential oils and GC-MS was used to confirm the identities of the isolated components. Preparative HPLC technique was used to separate and isolate the oils main components. The bulk EO and the separated components were tested for their antibacterial activities against Escherichia coli O157, Salmonella typhimurium, Klebsiella pneumonia, Yersinia enterocolitica O9, Brucella melitensis, Proteus spp., Pseudomonas aeruginosa, Staphylococcus aureus, Listeria monocytogenes and Bacillus cereus using broth microdilution method. The results confirm that the EOs and some of its components exhibited significant bactericidal activity. (author)

  10. Carboxymethylated chitosan-stabilized copper nanoparticles: a promise to contribute a potent antifungal and antibacterial agent

    International Nuclear Information System (INIS)

    Carboxymethylated chitosan (CMC)-stabilized copper nanoparticles (Cu-NPs) have been synthesized via chemical reduction of copper(II)–CMC complex in aqueous medium by hydrazine under microwave irradiation in ambient atmosphere. Structural morphology, phase, and chemical compositions of CMC-stabilized Cu-NPs (CMC–Cu-NPs) have been analyzed through high-resolution transmission electron microscopy, field emission scanning electron microscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. Antifungal and antibacterial activities of CMC–Cu-NPs have been evaluated against Candida tropicalis and Escherichia coli through agar well diffusion method, broth microdilution assay, live–dead assay, and microscopic observation. Antimicrobial activity of spherical CMC–Cu-NPs (∼4–15 nm of diameters) has been observed to be significant for both C. tropicalis and E. coli. The cytotoxicity study indicates that CMC–Cu-NPs have no significant toxic effect against normal cell line, L929

  11. Azole-synergistic anti-candidal activity of altenusin, a biphenyl metabolite of the endophytic fungus Alternaria alternata isolated from Terminalia chebula Retz.

    Science.gov (United States)

    Phaopongthai, Jatuporn; Wiyakrutta, Suthep; Meksuriyen, Duangdeun; Sriubolmas, Nongluksna; Suwanborirux, Khanit

    2013-12-01

    In this study, a tropical endophytic fungus, Alternaria alternata Tche-153 was isolated from a Thai medicinal plant Terminalia chebula Rezt. The ethyl acetate extract prepared from the fermentation broth exhibited significant ketoconazole-synergistic activity against Candida albicans. Bioassay-directed fractionation of the ethyl acetate extract led to the isolation of altenusin (1), isoochracinic acid (2), and altenuic acid (3) together with 2,5-dimethyl-7-hydroxychromone (4). Using the disc diffusion method and the microdilution chequerboard technique, only altenusin (1) in combination with each of three azole drugs, ketoconazole, fluconazole or itraconazole at their low sub-inhibitory concentrations exhibited potent synergistic activity against C. albicans with the fractional inhibitory concentration index range of 0.078 to 0.188. This first discovery of altenusin (1) as a new azole-synergistic prototype possessing a biphenyl structure is of significance for further development of new azole-synergists to treat invasive candidiasis. PMID:24385360

  12. Synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides: evaluation of antioxidant and antimicrobial activities

    Directory of Open Access Journals (Sweden)

    Juan I. Sarmiento-Sánchez

    2014-01-01

    Full Text Available A facile one-step synthesis of 1H-benzoxazine-2,4-diones from heterocyclic anhydrides and TMSA was described. This paper determines their antimicrobial activity against nine human bacterial pathogens by the broth microdilution method; antioxidant activity by DPPH• inactivation and a ferric-reducing power assay; and toxicity by a brine shrimp, Artemia salina, assay. The 1H-benzoxazine-2,4-dione yields were in the range of 57 to 98%. The novel compound 1H-pyrazino[2,3-][1,3]oxazine-2,4-dione 4c showed the highest antioxidant capacity (DPPH 35.4% and FRAP 0.063 µmol TEs/µmol.

  13. Management of Cosmetic Embarrassment Caused by Malassezia spp. with Fruticose Lichen Cladia Using Phylogenetic Approach

    Directory of Open Access Journals (Sweden)

    Anand Pandey

    2013-01-01

    Full Text Available During anti-Malassezia screening of plants by CLSI broth microdilution method, Cladia aggregata (Swartz Nyl. (family Cladoniaceae, a fruticose lichen from Sikkim (northeast Himalayan region, has been found effective at minimum inhibitory concentrations (mg/mL of 2.72, 0.63, and 1.28 against yeast-like fungi namely, M. furfur, M. globosa and M. sympodialis, respectively. These test pathogens are responsible for pityriasis versicolor (PV and seborrheic dermatitis (SD in humans. We tried to establish the reason for variable MICs against various Malassezia spp. using bioinformatical tools, thereby reducing the cost of the experimentation. This is the first report on anti-Malassezia activity of C. aggregata and thus can serve as a potential source for the development of cosmaceuticals.

  14. Ceftaroline activity tested against contemporary Latin American bacterial pathogens (2011

    Directory of Open Access Journals (Sweden)

    Robert K. Flamm

    2014-04-01

    Full Text Available A total of 2484 target bacterial pathogens were collected (one per patient episode from patients in 16 Latin American medical centers located in seven nations during 2011. Isolate identity was confirmed at a coordinating laboratory and susceptibility testing was performed for ceftaroline and comparator agents according to reference broth microdilution methods. A total of 30.0% of isolates were from respiratory tract, 29.4% from skin and skin structure, 21.4% from blood stream, 7.9% from urinary tract and 11.3% from other sites. Ceftaroline was active againstStaphylococcus aureus (42.8% MRSA with 83.6% of the isolates at 90.0% of the non-ESBL-phenotype. The spectrum of activity of ceftaroline against pathogens from Latin America indicates that it merits further study for its potential use in the Latin American region.

  15. Functional Ginger Extracts from Supercritical Fluid Carbon Dioxide Extraction via In Vitro and In Vivo Assays: Antioxidation, Antimicroorganism, and Mice Xenografts Models

    Directory of Open Access Journals (Sweden)

    Chih-Chen Lee

    2013-01-01

    Full Text Available Supercritical fluid carbon dioxide extraction technology was developed to gain the active components from a Taiwan native plant, Zingiber officinale (ginger. We studied the biological effects of ginger extracts via multiple assays and demonstrated the biofunctions in each platform. Investigations of ginger extracts indicated antioxidative properties in dose-dependant manners on radical scavenging activities, reducing powers and metal chelating powers. We found that ginger extracts processed moderate scavenging values, middle metal chelating levels, and slight ferric reducing powers. The antibacterial susceptibility of ginger extracts on Staphylococcus aureus, Streptococcus sobrinus, S. mutans, and Escherichia coli was determined with the broth microdilution method technique. The ginger extracts had operative antimicroorganism potentials against both Gram-positive and Gram-negative bacteria. We further discovered the strong inhibitions of ginger extracts on lethal carcinogenic melanoma through in vivo xenograft model. To sum up, the data confirmed the possible applications as medical cosmetology agents, pharmaceutical antibiotics, and food supplements.

  16. Chemical composition and antibacterial activity of the essential oil the leaf of Nepeta persica

    Directory of Open Access Journals (Sweden)

    Soraya AKHSHI

    2014-11-01

    Full Text Available The essential oil from the leaf of Nepeta persica Boiss, analyzed by gas chromatography (GC and gas chromatography (GC/mass spectrometry (MS, were shown to contain 4aα, 7α, 7aβ-nepetalactone (49.46% and 4aα, 7α, 7aα-nepetalactone (14.18%. The other main constituents were n-octane (13.10%, n-decane (3.67% and germacrene-D (2.04%. Antibacterial activities of the leaf oil were evaluated using the micro-dilution broth method. Inhibitory effects on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Enterococcus faecalis were recorded. The leaf oil has difference activities against the test microorganisms. The antibacterial property of the essential oil might be ascribed to their high content of nepetalactone isomers.

  17. In vitro assessment of the antimicrobial susceptibility of caprine isolates of Mycoplasma mycoides subsp. capri.

    Science.gov (United States)

    Paterna, A; Tatay-Dualde, J; Amores, J; Prats-van der Ham, M; Sánchez, A; de la Fe, C; Contreras, A; Corrales, J C; Gómez-Martín, Á

    2016-08-01

    The minimum inhibitory concentration (MIC) and minimum mycoplasmacidal concentration (MMC) of 17 antimicrobials against 41 Spanish caprine isolates of Mycoplasma mycoides subsp. capri (Mmc) obtained from different specimens (milk, external auricular canal and semen) were determined using a liquid microdilution method. For half of the isolates, the MIC was also estimated for seven of the antimicrobials using an epsilometric test (ET), in order to compare both methods and assess the validity of ET. Mutations in genes gyrA, gyrB, parC and parE conferring fluoroquinolone resistance, which have been recently described in Mmc, were investigated using PCR. The anatomical origin of the isolate had no effect on its antimicrobial susceptibility. Moxifloxacin and doxycycline had the lowest MIC values. The rest of the fluoroquinolones studied (except norfloxacin), together with tylosin and clindamycin, also had low MIC values, although the MMC obtained for clindamycin was higher than for the other antimicrobials. For all the aminoglycosides, spiramycin and erythromycin, a notable level of resistance was observed. The ET was in close agreement with broth microdilution at low MICs, but not at intermediate or high MICs. The analysis of the genomic sequences revealed the presence of an amino acid substitution in codon 83 of the gene gyrA, which has not been described previously in Mmc. PMID:27387734

  18. In vitro antifungal susceptibility of Malassezia furfur from bloodstream infections.

    Science.gov (United States)

    Iatta, Roberta; Figueredo, Luciana A; Montagna, Maria Teresa; Otranto, Domenico; Cafarchia, Claudia

    2014-11-01

    Fungaemia caused by Malassezia spp. in hospitalized patients requires prompt and appropriate therapy, but standard methods for the definition of the in vitro antifungal susceptibility have not been established yet. In this study, the in vitro susceptibility of Malassezia furfur from bloodstream infections (BSIs) to amphotericin B (AMB), fluconazole (FLC), itraconazole (ITC), posaconazole (POS) and voriconazole (VRC) was assessed using the broth microdilution (BMD) method of the Clinical and Laboratory Standards Institute (CLSI) with different media such as modified Sabouraud dextrose broth (SDB), RPMI and Christensen's urea broth (CUB). Optimal broth media that allow sufficient growth of M. furfur, and produce reliable and reproducible MICs using the CLSI BMD protocol were assessed. Thirty-six M. furfur isolates collected from BSIs of patients before and during AMB therapy, and receiving FLC prophylaxis, were tested. A good growth of M. furfur was observed in RPMI, CUB and SDB at 32 °C for 48 and 72 h. No statistically significant differences were detected between the MIC values registered after 48 and 72 h incubation. ITC, POS and VRC displayed lower MICs than FLC and AMB. These last two antifungal drugs showed higher and lower MICs, respectively, when the isolates were tested in SDB. SDB is the only medium in which it is possible to detect isolates with high FLC MICs in patients receiving FLC prophylaxis. A large number of isolates showed high AMB MIC values regardless of the media used. In conclusion, SDB might be suitable to determine triazole susceptibility. However, the media, the drug formulation or the breakpoints herein applied might not be useful for assessing the AMB susceptibility of M. furfur from BSIs. PMID:25168965

  19. A validated PCR-based method to detect Listeria monocytogenes using raw milk as a food model - Towards an international standard

    DEFF Research Database (Denmark)

    D'Agostino, M.; Wagner, M.; Vazquez-Boland, J.A.; Kuchta, T.; Karpiskova, R.; Hoorfar, Jeffrey; Novella, S.; Scortti, M.; Ellison, J.; Murray, A.; Fernandes, I.; Kuhn, M.; Pazlarova, J.; Heuvelink, A.; Cook, N.

    2004-01-01

    both the accordance (repeatability) and the concordance (reproducibility) were 99.4%. The assay was incorporated within a method for the detection of L. monocytogenes in raw milk, which involves 24 It of enrichment in half-Fraser broth followed by 16 h of enrichment in a medium that can be added...... concordance was 68.1%. The sensitivity (correct identification of milk samples inoculated with 20 to 200 L. monocytogenes cells per 25 ml) was 89.4%, the accordance was 81.2%, and the concordance was 80.7%. This method provides a basis for the application of routine PCR-based analysis to dairy products and...

  20. EFFICIENCY OF THE ENRICHMENT BROTHES (LEB1 AND LEB2 AND SELECTIVE AGARS (LPM AND LSAB-CAN TO ISOLATE BACTERIA OF THE GENUS Listeria IN MEAT AND RESIDUAL WATER OF WASHING CARCASS EFICIÊNCIA DOS CALDOS DE ENRIQUECIMENTO (LEB1 E LEB2 E DOS ÁGARES SELETIVOS (LPM E LSAB-CAN NO ISOLAMENTO DE BACTÉRIAS DO GÊNERO Listeria EM CARNE BOVINA E ÁGUA RESIDUÁRIA DE LAVAGEM DE CARCAÇA

    Directory of Open Access Journals (Sweden)

    Albenones José de Mesquita

    2007-09-01

    Full Text Available

    In this paper it was observed the efficiency of enrichment brothes for Listeria (LEB1 and LEB2, associated to the passage of the culture through a 0.25% potassium hidroxide solution, verifying that the secondary enrichment broth (LEB2 and LEB2KOH was better than the primary enrichment broth (LEB1KOH for this purpose. On the other hand, lithium chloride-phenylethanol-moxalactam agar and the selective Listeria agar base, supplemented with cicloheximide, acriflavine and nalidixic acid (LSAIB-CAN showed equivalency, at the statistic point of view (p=0.l442, in relation to the number of positive samples for Listeria spp., although the LSAB-CAN agar had given the greatest number of bacteria isolated from this genus.

    KEY-WORDS: Listeria; enrichment broth; selective agar.

    No presente trabalho verificou-se a eficiência dos caldos de enriquecimento para Listeria (LEB1 e LEB 2 associados à passagem da cultura por uma solução de hidróxido de potássio a 0,25%, constatando-se que o caldo de enriquecimento secundário (LEB2 e LEB2KOH foi superior ao caldo de enriquecimento primário (LEB1KOH. Por outro lado, o ágar cloreto de lítio-feniletanol-moxalactam (LPM e o ágar base seletivo para Listeria, suplementado com cicloheximida, acriflavina e ácido nalidíxico (LSAB-CAN equivaleram-se estatisticamente (p= 0,l442 em relação ao número de amostras positivas para Listeria, embora o ágar LSAB-CAN tenha proporcionado um maior número de isolamentos da bactéria.

    PALAVRAS-CHAVE: Caldo de enriquecimento; ágar seletivo; Listeria.

  1. Antibacterial activity against Clostridium genus and antiradical activity of the essential oils from different origin.

    Science.gov (United States)

    Kačániová, Miroslava; Vukovič, Nenad; Horská, Elena; Salamon, Ivan; Bobková, Alica; Hleba, Lukáš; Fiskelová, Martina; Vatľák, Alexander; Petrová, Jana; Bobko, Marek

    2014-01-01

    In the present study, the antimicrobial and antiradical activities of 15 essential oils were investigated. The antimicrobial activities were determined by using agar disc diffusion and broth microdilution methods against Clostridium genus and antioxidant properties of essential oils by testing their scavenging effect on DPPH radicals activities. We determined the antibacterial activity of Clostridium butyricum, Clostridium hystoliticum, Clostridium intestinale, Clostridium perfringens and Clostridium ramosum. We obtained the original commercial essential oils samples of Lavandula angustifolia, Carum carvi, Pinus montana, Mentha piperita, Foeniculum vulgare Mill., Pinus sylvestris, Satureia montana, Origanum vulgare L. (2 samples), Pimpinella anisum, Rosmarinus officinalis L., Salvia officinalis L., Abies alba Mill., Chamomilla recutita L. Rausch and Thymus vulgaris L. produced in Slovakia (Calendula a.s., Nova Lubovna, Slovakia). The results of the disk diffusion method showed very high essential oils activity against all tested strains of microorganisms. The best antimicrobial activity against C. butyricum was found at Pimpinella anisum, against C. hystoliticum was found at Pinus sylvestris, against C. intestinale was found at Satureia hortensis L., against C. perfringens was found at Origanum vulgare L. and against C. ramosum was found at Pinus sylvestris. The results of broth microdilution assay showed that none of the essential oils was active against C. hystoliticum. The best antimicrobial activity against C. butyricum was found at Abies alba Mill., against C. intestinale was found at Abies alba Mill., against C. perfringens was found at Satureia montana and against C. ramosum was found at Abius alba and Carum carvi. Antioxidant DPPH radical scavenging activity was determined at several solutions of oil samples (50 μL.mL(-1)-0.39 μL.mL(-1)) and the best scavenging effect for the highest concentration (50 μL.mL(-1)) was observed. The antioxidant properties

  2. A rapid method for the nonselective enumeration of Yersinia enterocolitica, a foodborne pathogen associated with pork.

    Science.gov (United States)

    Wang, Haoran; Palmer, Jon; Flint, Steve

    2016-03-01

    An impedance method was developed as a rapid, convenient method to enumerate pure culture of Yersinia enterocolitica. Cultures were incubated in trypticase soy broth (TSB) at 30°C. The BacTrac™ 4000 microorganism growth analyser was used to detect impedance change of TSB representing bacteria cell numbers in the samples. Good correlations with standard plate counts were obtained (r(2)>0.95). This method is also reliable to enumerate Y. enterocolitica growing in biofilms attached to stainless steel. Compared with a standard plate count which involves dislodging biofilms from surfaces, this method is more convenient saving both time and effort. Therefore, it will be useful to study the conditions required for the biofilm growth and control of Y. enterocolitica that could be applied to the pork industry. PMID:26613189

  3. Efficacy of Plectranthus amboinicus (Lour.) Spreng in a Murine Model of Methicillin-Resistant Staphylococcus aureus Skin Abscesses.

    Science.gov (United States)

    de Oliveira, Francisco Fábio Martins; Torres, Alba Fabiola; Gonçalves, Thially Braga; Santiago, Gilvandete Maria Pinheiro; de Carvalho, Cibele Barreto Mano; Aguiar, Milena Braga; Camara, Lilia Maria Carneiro; Rabenhorst, Silvia Helena; Martins, Alice Maria Costa; Valença Junior, José Telmo; Nagao-Dias, Aparecida Tiemi

    2013-01-01

    The present work aimed to evaluate the effectiveness of Plectranthus amboinicus (Lour.) Spreng against MRSA clinical isolates. The in vitro antimicrobial activity of the hydroalcoholic extract (HE), the ethyl acetate (EA) fraction and its subfractions were determined by broth microdilution and bioautography against MRSA clinical isolates. The microdilution checkerboard method was used to assess in vitro drug combination studies. To induce abscess formation, bacterial suspensions were added to Citodex and inoculated subcutaneously into male Swiss mice. The treatment protocol consisted of 2 doses of HE, the EA fraction or vancomycin introduced intraperitoneally into mice 3 and 12 h after infection. The EA fraction and its subfractions presented the lowest minimal inhibitory concentrations (MIC, 0.25 to 0.5 mg/mL). The plant samples were bacteriostatic at 2x and 4x MIC and bactericidal at 100 mg/mL. The EA fraction presented synergism with vancomycin and an additive effect with ciprofloxacin. A significant reduction of abscess volume, bacterial cell counts in abscess slurries, and inflammatory scores was observed in the HE and EA fraction-treated groups. The samples were effective in treating the animals in a dose-dependent fashion. The present study proved the effectiveness of P. amboinicus fractions against MRSA using in vitro and in vivo assays. PMID:23533472

  4. Efficacy of Plectranthus amboinicus (Lour. Spreng in a Murine Model of Methicillin-Resistant Staphylococcus aureus Skin Abscesses

    Directory of Open Access Journals (Sweden)

    Francisco Fábio Martins de Oliveira

    2013-01-01

    Full Text Available The present work aimed to evaluate the effectiveness of Plectranthus amboinicus (Lour. Spreng against MRSA clinical isolates. The in vitro antimicrobial activity of the hydroalcoholic extract (HE, the ethyl acetate (EA fraction and its subfractions were determined by broth microdilution and bioautography against MRSA clinical isolates. The microdilution checkerboard method was used to assess in vitro drug combination studies. To induce abscess formation, bacterial suspensions were added to Citodex and inoculated subcutaneously into male Swiss mice. The treatment protocol consisted of 2 doses of HE, the EA fraction or vancomycin introduced intraperitoneally into mice 3 and 12 h after infection. The EA fraction and its subfractions presented the lowest minimal inhibitory concentrations (MIC, 0.25 to 0.5 mg/mL. The plant samples were bacteriostatic at 2x and 4x MIC and bactericidal at 100 mg/mL. The EA fraction presented synergism with vancomycin and an additive effect with ciprofloxacin. A significant reduction of abscess volume, bacterial cell counts in abscess slurries, and inflammatory scores was observed in the HE and EA fraction-treated groups. The samples were effective in treating the animals in a dose-dependent fashion. The present study proved the effectiveness of P. amboinicus fractions against MRSA using in vitro and in vivo assays.

  5. Effect of Irradiation of Bacteria Upon Their Survival Rate During Conventional Methods of Meat Preservation

    International Nuclear Information System (INIS)

    The purpose of this paper is to define the effect of irradiation upon the survival rate of non-sporing bacteria (Staphylococcus aureus, Salmonella typhimurium, Escherichia coli, Pseudomonas fluorescens) during basic methods of meat preservation. The bacteria were irradiated in broth by X-rays at a dose that destroyed about 90% of the bacteria (D10). The survival rate of unirradiated and irradiated bacteria during cooling and freezing, in solutions of sodium chloride, nitrates and liquid smoke, was defined. The number of microorganisms was determined directly after irradiation as well as 1, 3, 7, 14, 21 and 28 days after irradiation. The effect of irradiation upon heat resistance of the examined species of bacteria was also defined. The microorganisms were heated in broth, at 70°C for 1, 2 and 5 minutes. The obtained results were subjected to statistical analysis. On the basis of the research results, a faster dying rate of irradiated populations of S. aureus and E. coli during any type of preservation treatment, the lack of any reaction to irradiation regarding the survival rate of S. typhimurium, and the lack of any effect of irradiation upon the rate of deterioration of P. fluorescens during freezing and storage in a solution with 10% addition of NaCl, were observed. On the other hand, a pronounced effect of irradiation upon the lowering of the heat resistance of the bacteria, as well as delayed growth in other variants of the experiment, was determined. (author)

  6. Effect of the irradiation of bacteria upon their survival rate during conventional methods of meat preservation

    International Nuclear Information System (INIS)

    The purpose of this paper is to define the effect of irradiation upon the survival rate of non-sporing bacteria (Staphylococcus aureus, Salmonella typhimurium, Escherichia coli, Pseudomonas fluorescens) during basic methods of meat preservation. The bacteria were irradiated in broth by X-rays at a dose that destroyed about 90% of the bacteria (D10). The survival rate of unirradiated and irradiated bacteria during cooling and freezing, in solutions of sodium chloride, nitrates and liquid smoke, was defined. The number of microorganisms was determined directly after irradiation as well as 1, 3, 7, 14, 21 and 28 days after irradiation. The effect of irradiation upon heat resistance of the examined species of bacteria was also defined. The microorganisms were heated in broth, at 700C for 1, 2 and 5 minutes. The obtained results were subjected to statistical analysis. On the basis of the research results, a faster dying rate of irradiated populations of S. aureus and E. coli during any type of preservation treatment, the lack of any reaction to irradiation regarding the survival rate of S. typhimurium, and the lack of any effect of irradiation upon the rate of deterioration of P. fluorescens during freezing and storage in a solution with 10% addition of NaCI, were observed. On the other hand, a pronounced effect of irradiation upon the lowering of the heat resistance of the bacteria, as well as delayed growth in other variants of the experiment, was determined. (author)

  7. [Comparison of ertapenem-EMB Agar with traditional methods for screening carbapenem-resistant Klebsiella pneumoniae from rectal swabs].

    Science.gov (United States)

    Perçin, Duygu; Colakoğlu, Selcan; Durmaz, Süleyman; Ekincioğlu, Pınar

    2012-10-01

    Detection of rectal colonization with carbapenem-resistant Klebsiella pneumoniae (CRKP) is the most important step in the infection control protocols in order to prevent infections caused by CRKP which has an increasing incidence all over the world. In this study, it was aimed to compare the detection rate of 2 mg/L ertapenem EMB agar medium with the other methods recommended by various international guidelines. These methods include direct plate method using ertapenem disc, enrichment method in tryptic soy broth containing 2 mg/L ertapenem and the investigation of the predominant betalactamases in the colonized patients. The lowest inoculum detected by different methods was determined by using simulative challenge test prepared for this purpose. The ability to detect CRKP from rectal swabs was evaluated by using the clinical specimens of 801 patients. For all bacteria isolated, carbapenem susceptibility was evaluated by using E-test method, the presence of beta-lactamases was determined by using modified Hodge test (MHT), and the carbapenemase genes were investigated by using multiplex polymerase chain reaction (PCR). The lowest inoculum detected by ertapenem-EMB agar was 50 CFU/mL whereas the lowest inocula were 1 x 105 and 1 x 103, respectively by tryptic soy broth with ertapenem and direct plate method. No resistance gene were identified by PCR in 13 (39.4%) of 33 isolates, whereas blaOXA-48 was detected in 19 (95%) and blaIMP in 1 (5%) of 20 positive isolates. All of the positive strains were resistant to imipenem and ertapenem, while 2 (10%) strains were found to be susceptible to doripenem and meropenem. While MHT was negative in all strains which were negative for resistance genes, all resistance gene positive strains except one blaOXA-48 strain that was also sensitive to doripenem and meropenem, were found to be positive with MHT. According to the results of PCR, the sensitivities of the three methods were found to be 80%. The specificities, positive and

  8. [A comparative study of various evaluation methods of the antibiotic sensitivity of strict anaerobic bacteria of the subgingival flora].

    Science.gov (United States)

    Kamagate, A; Kone, D; Coulibaly, N T; Brou, E; Sixou, M

    2001-09-01

    The study on the sensitiveness of slow-growing anaerobes bacteria to antibiotics is delicate when you consider the technical motives that make it difficult to transpose the standard methods frequently used in microbiological laboratories. The three main methods used to determine susceptibility to antibiotics are: disk-diffusion test, antibiotics containing microdilution plates and ATB ANA (bioMérieux). The aim of this study is to compare the effectiveness of each of these methods on severe anaerobes bacteria isolated in sub-gingival flora of patients suffering from developing periodontitis (rapidly progressive periodontitis, refractory periodontitis, active stage of adult chronic periodontitis). The observed bacteria are: Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Campylobacter rectus, Peptostreptococcus micros. Antibiotics used are: ampicilline, amoxicilline, tetracycline, erythromycine, metronidazole. The comparison of the minimal inhibitory concentrations (M.I.C) of each of these methods has permitted to show a strict correlation in the results observed with these three methods, if only the growth of the severe anaerobes bacteria on agar medium does not exceed 72 hours. PMID:11808376

  9. Antimicrobial Susceptibility of Udder Pathogens Isolated from Dairy Herds in the West Littoral Region of Uruguay

    Directory of Open Access Journals (Sweden)

    Franklin A

    2002-03-01

    Full Text Available A total of 522 strains belonging to streptococci, enterococci and staphylococci isolated from sub-clinical and clinical cases of bovine mastitis from the west littoral region of Uruguay were analysed for their susceptibility to several antimicrobial agents. The susceptibility patterns were studied by agar disk diffusion methods (ADDM and broth micro-dilution to determine the minimum inhibitory concentration (MIC. The concentration that inhibits 90% (MIC90 of the analysed strains reported in micrograms per millilitre, for Staphylococcus aureus were > 8, 8, ≤ 0.5, ≤ 4, ≤ 1, ≤ 0.5, > 64, ≤ 0.25, 0.5, ≤ 1 and ≤ 1 to penicillin, ampicillin, oxacillin, cephalotin, gentamicin, erythromycin, oxitetracycline, enrofloxacin, trimethoprim/sulfamethoxazole, neomycin, and clindamycin, respectively. Coagulase-negative staphylococci (CNS had different values for penicillin (4 and ampicillin (2, while the other antimicrobial agents had the same MIC90 values as reported for S. aureus. The MIC90 values for streptococci were 0.12, 0.25, ≤ 4, 16, ≤ 0.25, 0.5, 0.25 for penicillin, ampicillin, cephalotin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, whereas MIC90 for enterococci were 4, 4, 4, ≤ 0.5, 2, > 8 for penicillin, ampicillin, gentamicin, erythromycin, oxytetracycline and trimethoprim-sulfamethoxazole, respectively. Of 336 strains of S. aureus, 160 (47.6% were resistant to penicillin. For 41 CNS strains, 10 (27% presented penicillin-resistance. All the streptococcal strains were susceptible to penicillin, while 3 (7% of the 43 enteroccocal strains were resistant. Non significant statistical differences were found between the results obtained by ADDM and broth micro-dilution for classifying bacterial isolates as susceptible or resistant according to the National Committee of Clinical Laboratory Standards.

  10. [Detection and Serotyping of Streptococcus pneumoniae Carried in Healthy Adults with a Modified PCR Method].

    Science.gov (United States)

    Ishihara, Yuka; Okamoto, Akira; Ohta, Michio

    2015-05-01

    Detection of Streptococcus pneumoniae colonized in the pharynx of healthy carriers currently relies on conventional culture methods of direct plating with pharyngeal swab specimens. The accurate measurement of the carriage of pneumococci, however, has not been necessarily achieved with these methods due to low density colonization and contamination of numerous oral streptococci that express α-hemolysis. A PCR-based detection method of pneumococci-specific for lytA as well as PCR serotyping of S. pneumoniae was recently developed and their effectiveness was confirmed. We modified the reaction conditions of these methods to improve the detection rate and applied them to the measurement of S. pneumoniae carried in healthy adults. Pharyngeal swab specimens obtained from 110 healthy volunteers over 40 and living in Nagoya were enriched for 5 hours with broth medium supplemented with rabbit serum and the template DNA for PCR was extracted from the mixed enriched culture. Of 110 specimens 36 (32.7%) were lytA-positive, the rate of which was much higher than the results of previous culture-based studies. The DNA template preparations were then used for PCR-based serotyping with primers specific for each of the types included in pneumococcal 23 valent vaccine (PPV23). We found that 28 out of 36 lytA-positive carriers were identified as being positive for the serotypes belonging to PPV23, although serotypes 6A and 6B were indistinguishable with the PCR method. The most frequent serotype was serotype 14, and serotypes 4, 18C, and 6A/B were also frequently identified. Five lytA-positive carriers were previously vaccinated with PPV23, and among them, 4 were positive for serotypes contained in PPV23. We recommend PCR-based identification and serotyping of S. pneumoniae in broth enrichment culture of pharyngeal swab specimens as a reliable method for the surveillance of healthy carriers with low density colonization. PMID:26552129

  11. Evaluation of minimal inhibitory concentration of two new materials using tube dilution method: An in vitro study

    Directory of Open Access Journals (Sweden)

    Geeta S Hiremath

    2015-01-01

    Full Text Available Aim: The aim of the study is to evaluate and compare the antimicrobial efficacy of two new materials MTA Plus and Biodentine with ProRoot MTA using tube dilution method. Materials and Methods: The materials used were ProRoot MTA (Dentsply, MTA Plus (compounded by Prevest Denpro, Jammu, India for Avalon Biomed Inc, USA and a calcium silicate based material Biodentine (Septodont, Saint-Maur-des-Fosses, France. Doubling dilutions of the material were prepared in Sabouraud′s dextrose broth (SDB and Brain Heart Infusion (BHI broth for Candida albicans and Enterococcus faecalis, respectively. The minimal concentration at which inhibition of microorganism occurred was measured and noted as minimal inhibitory concentration (MIC of the material. Results: There was no statistically significant difference between the materials against C. albicans. Biodentine was statistically significant than MTA Plus against E. faecalis (P-value-0.022. ProRoot MTA was statistically significant at different time intervals against E. faecalis (P-value-0.001. Conclusion: ProRoot MTA and Biodentine proved to have antimicrobial property. MTA Plusproved as a good antifungal agent.

  12. In-situ product recovery from fermentation broths

    OpenAIRE

    Van den Berg, C.

    2010-01-01

    Life sciences will be one of the decisive factors in the 21st century. A major part of this field is the so-called white biotechnology, also known as industrial biotechnology. White biotechnology is an emerging field where bio-chemicals are produced using micro-organisms. However, the production of bio-chemicals is severely inhibited due to toxicity of the product towards the host-organism. Therefore, it is necessary to remove the products from the micro-organisms’ environment. One approach f...

  13. In-situ product recovery from fermentation broths

    NARCIS (Netherlands)

    Van den Berg, C.

    2010-01-01

    Life sciences will be one of the decisive factors in the 21st century. A major part of this field is the so-called white biotechnology, also known as industrial biotechnology. White biotechnology is an emerging field where bio-chemicals are produced using micro-organisms. However, the production of

  14. Optimization of Ultrafiltration Processes in Bolistin Fermentation Broth on the Base of Polyether Sulphone (PES) Memebrane%基于聚醚砜(PES)超滤膜的粘杆菌素发酵液预处理过程优化

    Institute of Scientific and Technical Information of China (English)

    魏安静; 田丽; 凤权

    2011-01-01

    Uhrafiltration was used to pretreat colistin fermentation broth. According to the characteristics of the material and comparison of their ultrafihration separation performance, polyether sulphone ultrafiltration membrane (MWCO = 10 000) was selected. The optimum operating parameters had been determined: pressure was 0.3 MPa, pH was 6.0. Under these optimum parameters, the protein rejected ratio was 96% , absorbency was 0. 185, and part of the pigment had been removed.%利用超滤对粘杆菌素发酵液进行膜分离处理并对其工艺过程进行优化。根据膜材料特性,选择聚醚砜(PES)为基材的超滤膜,通过不同截留分子量超滤膜的分离性能比较、操作条件选择,最终确定PES-10(截留分子质量为10000u)为超滤膜件,适宜的操作压力为0.3MPa、料液pH值为6。经过超滤处理,滤液蛋白质含量为4.6mg/100mL,蛋白质去除率为96%,吸光度为0.185,发酵液中的色素得到有效地去除。

  15. Effect of the fermentation broth of a bacteriocin-producing lactobacillus plantarum YJG on the production performance and egg quality of laying hens%一株产细菌素植物乳酸杆菌YJG发酵液对蛋鸡生产性能以及蛋品质的影响

    Institute of Scientific and Technical Information of China (English)

    韩冰; 邓凯; 张日俊

    2011-01-01

    To study effect of the fermentation broth of a bacteriocin-producing Lactobacillus plantarum YJG on production performance and egg quality of laying hens. Two hundred and twenty five laying hens of 232-day-old were randomly divided into 5 treatment groups with 5 replicates, and the trials period was 34 days. The results showed that treatment with 500 AU/kg bacteriocin could enhance the average weight, and the treatments with bacteriocins could apparently reduce the feed egg ratio(P<0.05), Bacteriocins produced by Lactobacillus plantarum YJG could improve the production performance and egg quality of laying hens and provide new insight into research of feed additives.%研究一株植物乳酸杆菌细菌素对蛋鸡生产性能和蛋品质的影响.选用232日龄产蛋率和体重相近、健康状况良好的海兰灰蛋鸡225只,试验期为34 d.结果表明:日粮中添加500 AU/kg植物乳酸杆菌细菌素对蛋鸡的产蛋率和平均蛋重的提升幅度最大,各细菌素添加组对降低料蛋比均有显著的效果(P<0.05),对平均日采食量、蛋黄色泽、哈氏单位、蛋壳强度和蛋形指数都有不同程度的改善.植物乳酸杆菌细菌素可以明显改善蛋鸡生产性能和蛋品质,可以为饲料添加剂的研发提供新的思路.

  16. Sterilization Effect of Wet Oxygen Plasma in the Bubbling Method.

    Science.gov (United States)

    Tamazawa, Kaoru; Shintani, Hideharu; Tamazawa, Yoshinori; Shimauchi, Hidetoshi

    2015-01-01

    A new low-temperature sterilization method to replace the ethylene oxide gas sterilization is needed. Strong bactericidal effects of OH and O2H radicals are well known. The purpose of this study was to evaluate the sterilization effect of wet oxygen ("O2+H2O") plasma in the bubbling method, confirming the effect of humidity. Sterility assurance was confirmed by using a biological indicator (Geobacillus stearothermophilus ATCC7953, Namsa, USA). One hundred and eight samples (10(5) spores/carrier) were divided into three groups of 36 in each for treatment with a different type of gas (O2, O2+H2O, Air+H2O). Plasma processing was conducted using a plasma ashing apparatus (13.56 MHz, PACK-3(®), Y. A. C., Japan) under various gas pressures (13, 25, 50 Pa) and gas flows (50, 100, 200 sccm). Fixed plasma treatment parameters were power at 150 W, temperature of 60 ℃, treatment time of 10 min. The samples after treatment were incubated in trypticase soy broth at 58 ℃ for 72 h. The negative culture rate in the "O2+H2O" group was significantly (Mantel-Haenszel procedure, psterilization effect of the "O2+H2O" group depends on the bubbling method which is the method of introducing vapor into the chamber. The bubbling method seems able to generate OH and O2H radicals in a stable way. PMID:26699857

  17. The evaluation of a PCR-based method for identification of Salmonella enterica serotypes from environmental samples and various food matrices.

    Science.gov (United States)

    Jean-Gilles Beaubrun, Junia; Cheng, Chorng-Ming; Chen, Kai-Shun; Ewing, Laura; Wang, Hua; Agpaoa, Maria C; Huang, Mei-Chiung J; Dickey, Erin; Du, Jamie M; Williams-Hill, Donna M; Hamilton, Brittany; Micallef, Shirley A; Rosenberg Goldstein, Rachel E; George, Ashish; Joseph, Sam W; Sapkota, Amy R; Jacobson, Andrew P; Tall, Ben D; Kothary, Mahendra H; Dudley, Kim; Hanes, Darcy E

    2012-09-01

    The most commonly used method for serotyping Salmonella spp. is based on the Kaufmann-White scheme, and is composed of serological reactions using antibodies to LPS agglutinins. The multiplex PCR used in this investigation was established by Kim et al. to serotype the 30 most common clinical Salmonella serotypes, as determined by CDC. The PCR assay consists of two five-plex reactions and a single two-plex PCR reaction, based on six genetic loci from Salmonella enterica serotype Typhimurium and four loci from S. enterica serotype Typhi. In this investigation, we further evaluated the method for serotyping Salmonella spp. using a reference collection, environmental samples collected from a Mid-Atlantic region tomato farm study, four food matrices spiked with different Salmonella serotypes and a proficiency test. The PCR assay was first evaluated using DNA isolated from pure cultures of isolates obtained from various clinical and environmental samples, and then DNA isolated from broth cultures of food matrices of "Red round" and Roma tomatoes, Romaine lettuce, green onions and Serrano peppers spiked with serotypes Newport, Typhimurium, Javiana and Saintpaul, respectively. The results showed that the PCR assay correctly serotyped Salmonella spp. from the clinical, environmental, spiked food matrices, and proficiency test samples. These findings are significant because the PCR assay was successful in the identification of Salmonella in the spiked samples in a broth culture containing other non-salmonella organism. This method may be a useful resource for the food safety community. PMID:22608224

  18. Occurrence and detection method evaluation of group B streptococcus from prenatal vaginal specimen in Northwest China.

    Science.gov (United States)

    Xie, Yun; Yang, JunLan; Zhao, Peng; Jia, Hui; Wang, Qi

    2016-01-01

    Sensitive and efficient detection of Group B Streptococcus (GBS) colonization in pregnant women is essential for prescription of prophylaxis at the time of delivery as GBS is an opportunistic pathogen known to cause infant mortality. In this report, two studies were conducted on the methods of GBS detection in Shaanxi province, China, a region lacking data for GBS detection and occurrence. For Study 1, 100 GBS culture-positive vaginal swabs were collected from 1,567 pregnant women for evaluation by direct latex agglutination test. In Study 2, 200 GBS vaginal swabs were evaluated by three culture methods (sheep blood agar (SBA), Columbia colistin-nalidixic agar (CNA), and selective carrot broth (SCB)) followed by analysis using a latex agglutination test. GBS was detected in 6.4 % of specimens in Study 1 and 10.5 % of specimens in Study 2. The results of the latex agglutination test in both studies were accurate with samples exhibiting high to moderate GBS growth, but the accuracy declined for samples with low GBS growth. The evaluation of culture methods for GBS detection revealed the sensitivity of SCB (95.2 %, p = 0.004) was significantly higher than that of the SBA medium (57.1 %). The sensitivity reported for SCB (95.2 %) was higher than CNA (76.0 %), but the difference was not statistically significant (p =0.078). These results indicate a selective broth, such as SCB, is ideal for accuracy at low growth levels, but a direct latex agglutination test could be used as an alternative for rapid detection of GBS in circumstances requiring immediate detection. PMID:26791082

  19. Effective disinfection methods of kitchen sponges

    Science.gov (United States)

    Pathogenic foodborne bacteria can be disseminated in households through the use of contaminated sponges. Several household disinfecting treatments to kill bacteria, yeasts and molds on sponges were evaluated. Sponges were incubated in a suspension of ground beef and tryptic soy broth to develop bact...

  20. Detection of pathogenic Yersinia enterocolitica in slaughtered pigs by cultural methods and real-time polymerase chain reaction

    Directory of Open Access Journals (Sweden)

    Rina Mazzette

    2015-05-01

    Full Text Available Healthy pigs carrying pathogenic to human Yersinia enterocolitica strains are the main source of entry into slaughterhouse, where cross-contamination of carcasses can happen. The aim of this work was to determine Y. enterocolitica prevalence in slaughtered pigs, investigating the presence of carriers in relation to carcass contamination. A total of 132 pig samples (tonsils, mesenteric lymph nodes, colon content, carcass surface were collected from 4 Sardinian slaughterhouses. All the samples were examined by the ISO 10273:2003 method, and the prevalence was also determined by direct plating on CIN Agar. Moreover, to detect the ail positive Y. enterocolitica strains in enrichment broths and isolates a real-time polymerase chain reaction (PCR was applied. Y. enterocolitica prevalence was 19% with direct plating and 12% with enrichment methods. Carcass surfaces and tonsils prevalence was 5.30% by direct plating, and 5.3% and 2.2%, respectively, by enrichment method. Tonsil samples showed an average contamination level of 3.2×103 CFU/g, while the mean value on carcass was 8.7×102 CFU/g. An overall prevalence of 9.8% of ail positive Y. enterocolitica broths was detected by RT-PCR, that found a higher prevalence in tonsils (7.5% with respect to cultural methods, confirming the greater sensitivity of this technique when applied for tonsils and faeces samples. The results show a relatively low pathogenic Y. enterocolitica prevalence in pigs slaughtered in Sardinia. Good hygiene measures should be applied at slaughterhouse in order to prevent the entry of carriers and control carcass contamination.

  1. Simple method for correct enumeration of Staphylococcus aureus.

    Science.gov (United States)

    Haaber, J; Cohn, M T; Petersen, A; Ingmer, H

    2016-06-01

    Optical density (OD) measurement is applied universally to estimate cell numbers of microorganisms growing in liquid cultures. It is a fast and reliable method but is based on the assumption that the bacteria grow as single cells of equal size and that the cells are dispersed evenly in the liquid culture. When grown in such liquid cultures, the human pathogen Staphylococcus aureus is characterized by its aggregation of single cells into clusters of variable size. Here, we show that aggregation during growth in the laboratory standard medium tryptic soy broth (TSB) is common among clinical and laboratory S. aureus isolates and that aggregation may introduce significant bias when applying standard enumeration methods on S. aureus growing in laboratory batch cultures. We provide a simple and efficient sonication procedure, which can be applied prior to optical density measurements to give an accurate estimate of cellular numbers in liquid cultures of S. aureus regardless of the aggregation level of the given strain. We further show that the sonication procedure is applicable for accurate determination of cell numbers using agar plate counting of aggregating strains. PMID:27080188

  2. 寡雄腐霉发酵液的动物毒性及其对柑橘果实贮藏期青、绿霉病的防治效果%Toxicity of Pythium oligandrum broth to animal and its control effect on rot diseases caused by Penicillium italicum and Penicillium digitatum in orange fruit storage

    Institute of Scientific and Technical Information of China (English)

    谭艳; 彭良志; 袁玲; 王少博

    2015-01-01

    [目的]研制安全、无毒、高效的生物保鲜剂,降低意大利青霉(Penicillium italicum,青霉)和指状青霉(Penicillium digitatum,绿霉)引起的柑橘烂果.[方法]试验利用自主选育的寡雄腐霉优良菌株(Pythium oligandrum CQ2010),制备发酵液,测试了对小鼠的急性毒性,并设置对照(液体培养基,CK)、寡雄腐霉发酵液(P.oligandrum Broth,POB)、咪鲜胺(Prochloraz,PC)、咪鲜胺+POB(PC+POB)等4种处理,研究了它们对青、绿霉菌的抑制作用及其对柑橘防腐保鲜的作用.[结果]用大剂量的POB灌胃给药对小鼠体重增长无显著影响,供试动物的外观和行为均无异常,心、肝、肾、肺、肠等组织器官也未见病理改变.POB显著抑制青、绿霉菌丝生长和孢子萌发,抑制率分别为70.24%-93.74%(菌丝生长)和44.91%-87.82% (24 h孢子萌发).柑橘果实接种青霉后,烂果率CK> POB、PC> PC+POB,防治效果PC+POB> POB、PC.在模拟柑橘商品化贮藏保鲜试验中,青、绿霉发病率占总发病率的50%以上,CK、POB、PC和PC+POB的烂果率依次为26.40%、15.03%、16.61%和4.21%.此外,POB对果实品质无显著影响,但显著提高果皮中的超氧化物歧化酶、过氧化物酶和过氧化氢酶活性,有益于提高柑橘果实的抗病性和贮藏性.[结论]在柑橘贮藏过程中,POB对果实青、绿霉病有显著的防治作用,并与咪鲜胺的防病效果有叠加作用.

  3. Field Evaluation of a Semiautomated Method for Rapid and Simple Analysis of Recreational Water Microbiological Quality

    Science.gov (United States)

    Anglès d'Auriac, Marc B.; Roberts, Hildegarde; Shaw, Terri; Sirevåg, Reidun; Hermansen, Leonila Fajardo; Berg, James D.

    2000-01-01

    An early warning system using a rapid enzymatic semiautomated method suitable for fecal coliform detection in recreational waters within 8 h was developed further and evaluated in this study. This rapid method was compared to the standard method followed in the United Kingdom. We used 1,011 samples originating from 206 different locations in Wales. When we assessed the presence or absence of fecal coliforms, targeting very low levels of contamination, we obtained 83.9% agreement between the rapid method and the lauryl sulfate broth-membrane filtration technique, whereas direct confirmation of the samples processed by the rapid method showed 89.3% agreement. Environmental enzymatic background activity was found to be the main limiting factor for this method. Owing to a specific and integrated handling of the results by the software of the instrument, the percentage of false-positive results (a consequence of enzymatic background) was successfully limited to 2.9% by the direct confirmation evaluation. However, 7.8% false-negative results due to “late-growers” had to be accepted in order to produce results within a working day. At present, the method can be used in a more conservative way to assess the environmental threshold of 100 CFU of fecal coliforms per 100 ml in recreational waters. The implications of our findings with regard to the applicability of rapid enzymatic methods are discussed. PMID:11010890

  4. Newton's method

    Energy Technology Data Exchange (ETDEWEB)

    More, J. J.; Sorensen, D. C.

    1982-02-01

    Newton's method plays a central role in the development of numerical techniques for optimization. In fact, most of the current practical methods for optimization can be viewed as variations on Newton's method. It is therefore important to understand Newton's method as an algorithm in its own right and as a key introduction to the most recent ideas in this area. One of the aims of this expository paper is to present and analyze two main approaches to Newton's method for unconstrained minimization: the line search approach and the trust region approach. The other aim is to present some of the recent developments in the optimization field which are related to Newton's method. In particular, we explore several variations on Newton's method which are appropriate for large scale problems, and we also show how quasi-Newton methods can be derived quite naturally from Newton's method.

  5. Evaluation of the Antimicrobial Activity of the K9CATH Peptide (38 Amino Acids Against a Mastitis Isolated Strain of Staphylococcus aureus by the Resazurin microtiter Method

    Directory of Open Access Journals (Sweden)

    Albero Barreras-Serrano

    2014-04-01

    Full Text Available The antimicrobial activity of the synthetic peptide K9CATH was determined by the Resazurin microtitre Method (RMM against a strain of S. aureus isolated from a case of mastitis. To the antibiogram this bacteria strain showed to be resistant to Ampicillin, Erythromycin, Cefeprime, Dicloxaciline and Penicillin (10 U, while the MIC obtained for the K9CATH was 5.66 &mug/mL. Unlike the reference broth method, visual reading for MIC determination with the RMM showed to be easier, rapid, inexpensive and more sensitive for antimicrobial peptide screening, based in a color change from blue (not growth to pink (growth. This is the first time that the resazurin method is used to determine the MIC of the 38 aa´s K9CATH peptide against a mastitic isolate of S. aureus.

  6. Low minimum inhibitory concentrations associated with the tetracycline-resistance gene tet(C) in Escherichia coli

    OpenAIRE

    Chalmers, Gabhan; Kozak, Gosia K.; Hillyer, Elizabeth; Reid-Smith, Richard J.; Boerlin, Patrick

    2010-01-01

    Twenty-eight Escherichia coli isolates from various animal and environmental sources with defined tetracycline-resistance genotypes for tet(A), tet(B), and tet(C) were tested for their susceptibility to tetracycline by means of both broth microdilution and Etest. All tet(C)-positive isolates had tetracycline minimum inhibitory concentrations clustering around an intermediate susceptibility range of 2 to 16 μg/mL. Detecting tet(C)-positive isolates by means of susceptibility testing may theref...

  7. Rifaximin disc diffusion test for in vitro susceptibility testing of Clostridium difficile

    OpenAIRE

    Huhulescu, Steliana; Sagel, Ulrich; Fiedler, Anita; Pecavar, Verena; Blaschitz, Marion; Wewalka, Guenther; Allerberger, Franz; Indra, Alexander

    2011-01-01

    Rifaximin is a rifampicin derivative, poorly absorbed by the gastro-intestinal tract. We studied the in vitro susceptibility to rifamixin of 1082 Clostridium difficile isolates; among these,184 isolates from a strain collection were tested by an in-house rifaximin disc (40 µg) diffusion test, by an in-house rifaximin broth microdilution test, by rifampicin Etest and by rpoB gene sequencing. In the absence of respective CLSI or EUCAST MIC breakpoints for rifaximin and rifampicin against C. dif...

  8. Complete genome sequences of three Pseudomonas aeruginosa isolates with phenotypes of polymyxin B adaptation and inducible resistance.

    Science.gov (United States)

    Boyle, Brian; Fernandez, Lucia; Laroche, Jerome; Kukavica-Ibrulj, Irena; Mendes, Caio M F; Hancock, Robert W; Levesque, Roger C

    2012-01-01

    Clinical "superbug" isolates of Pseudomonas aeruginosa were previously observed to be resistant to several antibiotics, including polymyxin B, and/or to have a distinct, reproducible adaptive polymyxin resistance phenotype, identified by observing "skipped" wells (appearance of extra turbid wells) during broth microdilution testing. Here we report the complete assembled draft genome sequences of three such polymyxin resistant P. aeruginosa strains (9BR, 19BR, and 213BR). PMID:22207740

  9. Complete Genome Sequences of Three Pseudomonas aeruginosa Isolates with Phenotypes of Polymyxin B Adaptation and Inducible Resistance

    OpenAIRE

    Boyle, Brian; Fernandez, Lucia; Laroche, Jerome; Kukavica-Ibrulj, Irena; Caio M. F. Mendes; Hancock, Robert W.; Levesque, Roger C.

    2012-01-01

    Clinical “superbug” isolates of Pseudomonas aeruginosa were previously observed to be resistant to several antibiotics, including polymyxin B, and/or to have a distinct, reproducible adaptive polymyxin resistance phenotype, identified by observing “skipped” wells (appearance of extra turbid wells) during broth microdilution testing. Here we report the complete assembled draft genome sequences of three such polymyxin resistant P. aeruginosa strains (9BR, 19BR, and 213BR).

  10. GC and GC/MS Analysis of Essential Oil Composition of the Endemic Soqotraen Leucas virgata Balf.f. and Its Antimicrobial and Antioxidant Activities

    OpenAIRE

    Ramzi A. Mothana; Mansour S. Al-Said; Al-Yahya, Mohammed A.; Al-Rehaily, Adnan J; Jamal M. Khaled

    2013-01-01

    Leucas virgata Balf.f. (Lamiaceae) was collected from the Island Soqotra (Yemen) and its essential oil was obtained by hydrodistillation. The chemical composition of the oil was investigated by GC and GC-MS. Moreover, the essential oil was evaluated for its antimicrobial activity against two Gram-positive bacteria, two Gram-negative bacteria, and one yeast species by using broth micro-dilution assay for minimum inhibitory concentrations (MIC) and antioxidant activity by measuring the scavengi...

  11. Antimicrobial susceptibility testing of less commonly isolated Haemophilus species using Haemophilus test medium.

    OpenAIRE

    Jorgensen, J H; Howell, A W; Maher, L A

    1990-01-01

    Haemophilus test medium (HTM) was developed recently for dilution and disk diffusion antimicrobial agent susceptibility testing of Haemophilus influenzae. The application of HTM to the testing of other, less frequently encountered Haemophilus species recovered from humans was evaluated in this study by using commercially prepared HTM (BBL Microbiology Systems, Cockeysville, Md.) in broth microdilution and agar disk diffusion susceptibility tests with 18 antimicrobial agents. A total of 93.3% ...

  12. In Vitro Interactions between Target of Rapamycin Kinase Inhibitor and Antifungal Agents against Aspergillus Species.

    Science.gov (United States)

    Gao, Lujuan; Ding, Xiaozhen; Liu, Zhun; Wu, Qingzhi; Zeng, Tongxiang; Sun, Yi

    2016-06-01

    In vitro interactions of INK128, a target of rapamycin (TOR) kinase inhibitor, and antifungals, including itraconazole, voriconazole, posaconazole, amphotericin B, and caspofungin, against Aspergillus spp. were assessed with the broth microdilution checkerboard technique. Our results suggested synergistic effects between INK128 and all azoles tested, against multiple Aspergillus fumigatus and Aspergillus flavus isolates. However, no synergistic effects were observed when INK128 was combined with amphotericin B or caspofungin. No antagonism was observed for any combination. PMID:26976874

  13. Antimicrobial susceptibilities of clinical isolates of Acinetobacter baumannii from Singapore.

    OpenAIRE

    Kuah, B G; Kumarasinghe, G; Doran, J.; Chang, H R

    1994-01-01

    The in vitro activities of 17 antimicrobial agents alone or in combination against 70 clinical isolates of Acinetobacter baumannii from Singapore were determined by broth microdilution. The MICs of amoxicillin, ampicillin, ceftazidime, ceftriaxone, gentamicin, and piperacillin for 90% of the strains were > or = 128 micrograms/ml. Addition of sulbactam to ampicillin produced improved activity, whereas adding tazobactam to piperacillin did not. The MICs of amikacin, ciprofloxacin, and imipenem ...

  14. In vitro antimicrobial susceptibility of Mycoplasma mycoides mycoides large colony and Arcanobacterium pyogenes isolated from clinical cases of ulcerative balanitis and vulvitis in Dorper sheep in South Africa

    OpenAIRE

    A. Kidanemariam; J. Gouws; M. Van Vuuren; Gummow, B.

    2005-01-01

    The in vitro activities of enrofloxacin, florfenicol, oxytetracycline and spiramycin were determined against field isolates of Mycoplasma mycoides mycoides large colony (MmmLC) by means of the broth microdilution technique. The minimum inhibitory concentrations (MICs) of these antimicrobial drugs were determined for a representative number of 10 isolates and 1 type strain. The susceptibility of Arcanobacterium pyogenes to enrofloxacin, oxytetracycline and tilmicosin was determined by means of...

  15. In vitro antibiotic susceptibility of Dutch Mycoplasma synoviae field isolates originating from joint lesions and the respiratory tract of commercial poultry

    OpenAIRE

    Landman, Wil J.M.; Mevius, Dik J.; Veldman, Kees T.; Feberwee, Anneke

    2008-01-01

    Abstract The in vitro susceptibility of 17 Dutch Mycoplasma synoviae field isolates, 3 originating from joint pathology and 14 from the respiratory tract of commercial poultry, for enrofloxacin, difloxacin, doxycycline, tylosin and tilmicosin was examined. The M. synoviae ATCC 25204 was included as a control strain. The antibiotic susceptibility was tested quantitatively using the broth microdilution test. Based on initial and final MIC values, all tested isolates were susceptible ...

  16. EVALUATION OF ANTIMICROBIAL AND CYTOTOXIC ACTIVITIES OF PLANT EXTRACTS FROM SOUTHERN MINAS GERAIS CERRADO

    OpenAIRE

    Juliana Moscardini Chavasco; Barbara Helena Muniz Prado E Feliphe; Claudio Daniel Cerdeira; Fabricio Damasceno Leandro; Luiz Felipe Leomil Coelho; Jeferson Junior da Silva; Jorge Kleber Chavasco; Amanda Latercia Tranches Dias

    2014-01-01

    The antimicrobial activity of plant hidroethanolic extracts on bacteria Gram positive, Gram negative, yeasts, Mycobacterium tuberculosis H37 and Mycobacterium bovis was evaluated by using the technique of Agar diffusion and microdilution in broth. Among the extracts evaluated by Agar diffusion, the extract of Bidens pilosa leaf presented the most expressive average of haloes of growth inhibition to the microorganisms, followed by the extract of B. pilosa flower, of Eugenia pyriformis' leaf an...

  17. Re-Examining the Role of Hydrogen Peroxide in Bacteriostatic and Bactericidal Activities of Honey

    OpenAIRE

    Brudzynski, Katrina; Abubaker, Kamal; St-Martin, Laurent; Castle, Alan

    2011-01-01

    The aim of this study was to critically analyze the effects of hydrogen peroxide on growth and survival of bacterial cells in order to prove or disprove its purported role as a main component responsible for the antibacterial activity of honey. Using the sensitive peroxide/peroxidase assay, broth microdilution assay and DNA degradation assays, the quantitative relationships between the content of H2O2 and honey’s antibacterial activity was established. The results showed that: (A) the average...

  18. Antifungal susceptibility testing of isolates from a randomized, multicenter trial of fluconazole versus amphotericin B as treatment of nonneutropenic patients with candidemia. NIAID Mycoses Study Group and the Candidemia Study Group.

    OpenAIRE

    Rex, J. H.; Pfaller, M. A.; Barry, A L; Nelson, P W; Webb, C. D.

    1995-01-01

    The antifungal susceptibilities of 232 pathogenic blood stream Candida isolates collected during a recently completed trial comparing fluconazole (400 mg/day) with amphotericin B (0.5 mg/kg of body weight per day) as treatment for candidemia in the nonneutropenic patient were determined both by the National committee for Clinical Laboratory Standards M27-P macrobroth methodology and by a less cumbersome broth microdilution methodology. For amphotericin B, M27-P yielded a very narrow range of ...

  19. Antimicrobial activities of essential oils and crude extracts from tropical Citrus spp. against food-related microorganisms

    OpenAIRE

    Tipparat Hongpattarakere; Suphitchaya Chanthachum; Sumonrat Chanthaphon

    2008-01-01

    Ethyl acetate extracts and hydrodistillated-essential oils from peels of Citrus spp. were investigated for their antimicrobial activities against food related microorganisms by broth microdilution assay. Overall, ethyl acetate extracts from all citrus peels showed stronger antimicrobial activities than their essential oils obtained from hydrodistillation. The ethyl acetate extract of kaffir lime (Citrus hystrix DC.) peel showed broad spectrum of inhibition against all Gram-positive bacteria, ...

  20. Antifungal effect of cumin essential oil alone and in combination with antifungal drugs

    OpenAIRE

    SAHADEO PATIL; PANKAJ MAKNIKAR; SUSHILKUMAR WANKHADE; CHANDRAKIRAN UKESH; MAHENDRA RA

    2015-01-01

    Abstract. Patil S, Maknikar P, Wankhade S, Ukesh C, Rai M. 2015. Antifungal effect of cumin essential oil alone and in combination with antifungal drugs. Nusantara Bioscience 7: 55-59. We report evaluation of antifungal activity of cumin seed oil and its pharmacological interactions when used in combination with some of the widely used conventional antifungal drugs using CLSI broth microdilution, agar disc diffusion and checkerboard microtitre assay against Candida. The essential oil was obta...

  1. Chemical Composition, Antimicrobial and Antioxidant Activities of the Volatile Oil of Ganoderma pfeifferi Bres

    OpenAIRE

    Mohamed Al-Fatimi; Martina Wurster; Ulrike Lindequist

    2016-01-01

    In a first study of the volatile oil of the mushroom basidiomycete Ganoderma pfeifferi Bres., the chemical composition and antimicrobial and antioxidant activities of the oil were investigated. The volatile oil was obtained from the fresh fruiting bodies of Ganoderma pfeifferi Bres. By hydrodistillation extraction and analyzed by GC-MS. The antimicrobial activity of the oil was evaluated against five bacteria strains and two types of fungi strains, using disc diffusion and broth microdilution...

  2. In Vitro Activities of Cethromycin (ABT-773), a New Ketolide, against Streptococcus pneumoniae Strains That Are Not Susceptible to Penicillin or Macrolides

    OpenAIRE

    Mason, Edward O.; Lamberth, Linda B.; Wald, Ellen R.; Bradley, John S.; Barson, William J.; Kaplan, Sheldon L.

    2003-01-01

    Pneumococcal resistance to antimicrobials presents problems to physicians for empirical treatment of acute otitis media (AOM). Three hundred thirty-three isolates of Streptococcus pneumoniae selected for nonsusceptibility to penicillin (MIC >0.1 μg/ml) from the middle ear (n = 325) or mastoid (n = 8) of children seen between 1994 and 2000 at four children's hospitals in the United States were tested by broth microdilution for susceptibility to nine antibiotics. Using NCCLS 2002 breakpoints, r...

  3. New trends in dentistry: plant extracts against Enterococcus faecalis. The efficacy compared to chlorhexidine

    OpenAIRE

    Adriana Lígia de Castilho; Cintia Helena Coury Saraceni; Ingrit Elida Collantes Díaz; Mateus Luís Barradas Paciencia; Ivana Barbosa Suffredini

    2013-01-01

    Enterococcus faecalis is an important pathogen associated with endodontic diseases, and its elimination and control are of paramount importance, as it represents one of the major causes of failure in the treatment of endodontic disease. Twenty-five plant extracts obtained from Brazilian forests were found to be effective against planktonic E. faecalis and were subjected to two traditional antibacterial assays, the microdilution broth assay (MDBA) and the disk diffusion assay (DDA), using chlo...

  4. Ciprofloxacin-resistant methicillin-resistant Staphylococcus aureus in an acute-care hospital.

    OpenAIRE

    Raviglione, M. C.; Boyle, J. F.; Mariuz, P; Pablos-Mendez, A; Cortes, H; Merlo, A.

    1990-01-01

    Use of ciprofloxacin as an alternative to vancomycin for treatment of methicillin-resistant Staphylococcus aureus infection has been paralleled by the emergence of resistant strains. This phenomenon has also been noticed in our hospital. To confirm our observation, methicillin and ciprofloxacin susceptibilities were tested by disk diffusion and broth microdilution techniques. We studied 83 methicillin-resistant Staphylococcus aureus isolates obtained from various sources over a 4-month period...

  5. QUANTIFICATION OF LISTERIA MONOCYTOGENES IN MILK BY MPN-PCR AND MPN-CULTURE METHODS

    Directory of Open Access Journals (Sweden)

    Mahzad Hosseini

    2014-10-01

    Full Text Available The aim of this study was to compare the MPN-PCR (Most Probable Number- Polymerase Chain Reaction and MPN-Culture methods in enumerating of Listeria monocytogenes in milk. In order to compare the accuracy of these methods, 103 cell/ml Listeria monocytogenes and different background bacteria which may be present in raw milk, were inoculated in sterilized milk. After preparing serial dilutions, three replicates per dilution were inoculated in tubes containing listeria enrichment broth. After 48 hours of incubation, for MPN-Culture three inoculated replicates were subcultured on Oxford agar and suspected colonies were confirmed by performing by biochemical tests. For MPN-PCR assay, the DNA extraction was performed from the three inoculated replicates which were already used for MPN-Culture and PCR assay was performed using primers specific for Listeria monocytogenes. The experiment was repeated three times and the average of enumerated bacteria was calculated by each method separately. Statistical analysis using one sample Wilcoxon signed rank test showed that enumeration by MPN-PCR method was more accurate than enumeration by MPN-Culture method. The result of this study showed that MPN-PCR method in comparision with MPN-Culture even in the presence of different background microorganisms is more rapid and reliable. It is concluded that MPN-PCR method facilitates the enumeration of Listeria monocytogenes without excessive work and could be considered as an alternative to MPN-Culture technique.

  6. Fermentation, fractionation and purification of streptokinase by chemical reduction method

    Directory of Open Access Journals (Sweden)

    M Niakan

    2011-05-01

    Full Text Available Background and Objectives: Streptokinase is used clinically as an intravenous thrombolytic agent for the treatment of acute myocardial infarction and is commonly prepared from cultures of Streptococcus equisimilis strain H46A. The objective of the present study was the production of streptokinase from strain H46A and purification by chemical reduction method."nMaterials and Methods: The rate of streptokinase production evaluated under the effect of changes on some fermentation factors. Moreover, due to the specific structure of streptokinase, a chemical reduction method employed for the purification of streptokinase from the fermentation broth. The H46A strain of group C streptococcus, was grown in a fermentor. The proper pH adjusted with NaOH under glucose feeding in an optimum temperature. The supernatant of the fermentation product was sterilized by filtration and concentrated by ultrafiltration. The pH of the concentrate was adjusted, cooled, and precipitated by methanol. Protein solution was reduced with dithiothreitol (DTT. Impurities settled down by aldrithiol-2 and the biological activity of supernatant containing streptokinase was determined."nResults: In the fed -batch culture, the rate of streptokinase production increased over two times as compared with the batch culture and the impurities were effectively separated from streptokinase by reduction method."nConclusion: Improvements in SK production are due to a decrease in lag phase period and increase in the growth rate of logarithmic phase. The methods of purification often result in unacceptable losses of streptokinase, but the chemical reduction method give high yield of streptokinase and is easy to perform it.

  7. Use of radioactive ethanolamine incorporation into phospholipids to assess in vitro antimalarial activity by the semiautomated microdilution technique.

    OpenAIRE

    Elabbadi, N; Ancelin, M.L.; Vial, H J

    1992-01-01

    Phospholipid biosynthetic activity is intense in the erythrocytic stage of Plasmodium falciparum because of the parasite's own enzymatic machinery. The incorporation of various labeled phospholipid precursors in comparison with the incorporation of nucleic acid and protein precursors was tested to evaluate P. falciparum growth in vitro. These precursors, namely, [3H]ethanolamine, [3H]hypoxanthine, [3H]palmitate, [14C]serine, [3H]choline, [3H]inositol, and [3H]isoleucine, were all accurate ind...

  8. Sampling methods

    OpenAIRE

    Kılıç, Selim

    2013-01-01

    Most of the studies related to health are conducted with samples. Selection of sampling may be performed with random or nonrandom sampling methods. Sampling methods may vary with respect to some characteristics of study population, researchers’ aims and facilities. It is necessary to select samples by random sampling methods in order to provide the representativeness of the study population. The advantages and disadvantages of different sampling methods are presented in this manuscript which ...

  9. Studies on bactericidal efficacy of pumpkin (Cucurbita moschata Duchesne peel

    Directory of Open Access Journals (Sweden)

    El Zawane Kamarudin

    2014-02-01

    Full Text Available Objective: T o explore the in vitro antibacterial potential of the peel of Cucurbita moschata D uchesne ( tropical pumpkin ( C. moschata against human pathogenic bacteria. Methods: I n the present study, dichloromethane ( DCM , methanol ( MEOH and aqueous extracts of C. moschata peel were examined for in vitro antibacterial potency against eight bacterial strains i.e. Bacillus cereus, Burkholderia cepacia, Escherichia coli, Enterococcus faecalis, Staphyloccocus aureus, Pseudomonas aerugenosa, Vibrio alginolyticus, Vibrio parahaemolyticus using K irby- B auer disk diffusion susceptibility and broth micro-dilution methods. Results: DCM extract of pumpkin peel exhibited the maximum zone of inhibition against Staphyloccocus aureus ( 21 mm whereas aqueous extract of pumpkin peel revealed the least zone of inhibition against Escherichia coli ( 8 mm . MEOH extract gave maximum zone of inhibition against Pseudomonas aerugenosa ( 19 mm . B roth micro-dilution method showed minimum inhibitory concentration for the DCM extract against Burkholderia cepacia at 6 . 25 mg/m L . T he minimum bactericidal concentrations were also determined to know the nature of all extracts. DCM and MEOH extracts exhibited bactericidal nature to all bacterial strains except for the Vibrio alginolyticus. T he minimum bactericidal concentrations values exhibited bactericidal nature ranging from 3 . 12 mg/m L to 100 . 00 mg/m L . T he screening of antimicrobial properties of different extracts of C. moschata peel revealed that the DCM extract possessed good antimicrobial efficacy compared to MEOH and aqueous extracts. Conclusions: P eel of C. moschata possesses antibacterial compounds and could be potential source for a new class of antibiotics.

  10. Antimicrobial activity of linezolid combined with minocycline against vancomycin-resistant Enterococci

    Institute of Scientific and Technical Information of China (English)

    WU Jing; JIANG Tian-tong; SU Jian-rong; LI Li

    2013-01-01

    Background Vancomycin-resistant Enterococci (VRE) cause serious infections that are difficult to treat.We carried out this study to determine the mutant prevention concentration (MPC) of linezolid when combined with minocycline against VRE strains,to determine the mechanism of drug resistance in vitro,and to provide a theoretical basis for the rational use of drugs against VRE.Methods The minimum inhibitory concentrations (MICs) of linezolid and minocycline against 30 Enterococci (E.) isolates (including 20 VRE strains) were determined by the broth microdilution method.Drug interactions were assessed by the checkerboard microdilution tests and confirmed by time-kill studies.Two vancomycin-susceptible strains N27 and N40 (linezolid MIC,2 g/ml; minocycline MIC,4 μg/ml) and control strains E.faecalis ATCC 29212 and ATCC 51299 were also tested.The MPCs of linezolid and minocycline (alone and combined) were determined using the agar dilution method.Strains showing stable resistance were analyzed by polymerase chain reaction (PCR) amplification of domain V of the 23S rRNA gene.Results Checkerboard titration studies revealed synergistic effects of combination therapy in 26.7% of 30 E.isolates.Antagonism was not observed.The G2576U mutation was detected in stable linezolid-resistant strains of ATCC 29212,N40,and N27 before and after resistance screening,and MIC values increased with the number of G2576U mutations.The MPC of linezolid against E.decreased dramatically when combined with minocycline,and vice versa.Conclusion Linezolid or minocycline alone produce resistant strains; however,their joint use may reduce the MPC of each agent against VRE,thereby decreasing resistant mutants and bacterial infections.

  11. Screening Togolese medicinal plants for few pharmacological properties

    Directory of Open Access Journals (Sweden)

    Simplice D Karou

    2012-01-01

    Full Text Available Background: Terminalia macroptera Guill. et Perr. (Combretaceae, Sida alba L. (Malvaceae, Prosopis africana Guill et Perr. Taub. (Mimosaceae, Bridelia ferruginea Benth. (Euphorbiaceae, and Vetiveria nigritana Stapf. (Asteraceae are traditionally used in Togolese folk medicine to treat several diseases including microbial infections. Objective: This study aimed to investigate the antimicrobial, antioxidant, and hemolytic properties of the crude extracts of the above-mentioned plants. Materials and Methods: The antimicrobial and the antioxidant activities were assayed using the NCCLS microdilution method and the DPPH free radical scavenging, respectively. Human A+ red blood cells were used to perform the hemolytic assay. Phenolics were further quantified in the extracts using spectrophotometric methods. Results: Minimal inhibitory concentrations in the range of 230-1800 μg/ml were recorded in the NCCLS broth microdilution for both bacterial and fungal strains with methanol extracts. The DPPH radical scavenging assay yielded interesting antioxidant activities of the extracts of P. africana and T. macroptera (IC 50 values of 0.003 ± 0.00 μg/ml and 0.05 ± 0.03 μg/ml, respectively. These activities were positively correlated with the total phenolic contents and negatively correlated with the proanthocyanidin content of the extracts. The hemolytic assay revealed that great hemolysis occurred with the methanol extracts of T. macroptera, S. longepedunculata, and B. ferruginea. Conclusion: These results support in part the use of the selected plants in the treatment of microbial infections. In addition, the plant showed an interesting antioxidant activity that could be useful in the management of oxidative stress.

  12. Comparative studies on the activity of basil--an essential oil from Ocimum basilicum L.--against multidrug resistant clinical isolates of the genera Staphylococcus, Enterococcus and Pseudomonas by using different test methods.

    Science.gov (United States)

    Opalchenova, G; Obreshkova, D

    2003-07-01

    The essential oil basil is obtained from the aerial parts of Ocimum basilicum L. After gas chromatographic separation, the following components were identified: linalol (54.95%), methylchavikol (11.98%), methylcinnamat (7.24%) and linolen (0.14%). The activity of basil against multidrug resistant clinical isolates from the genera Staphylococcus, Enterococcus and Pseudomonas has been studied. For this purpose, standard and modified broth macrodilution methods were used and time kill kinetic of basil was studied. The minimum inhibitory concentrations (MICs) were reported between 0.0030% and 0.0007% (v/v). These concentrations were compared with the inhibitory concentrations (ICs) and the logs of the bacterial counts reduction both obtained by basil diluted in 1% Tween (Tw) 80, saline test solution (STS) and spiritus vini (Sv) 95 degrees instead in a broth. The data, obtained after application of different methods of investigation and validated with membrane filtration, showed a strong inhibitory effect of basil on the test bacteria. The chosen bacteria are widespread and pose serious therapeutic difficulties because of their high level of resistance. For this reason, the results obtained were considered encouraging. PMID:12732427

  13. Projection Methods

    DEFF Research Database (Denmark)

    Wagner, Falko Jens; Poulsen, Mikael Zebbelin

    1999-01-01

    When trying to solve a DAE problem of high index with more traditional methods, it often causes instability in some of the variables, and finally leads to breakdown of convergence and integration of the solution. This is nicely shown in [ESF98, p. 152 ff.].This chapter will introduce projection...... methods as a way of handling these special problems. It is assumed that we have methods for solving normal ODE systems and index-1 systems....

  14. Discipline methods

    OpenAIRE

    Maria Kikila; Ioannis Koutelekos

    2012-01-01

    Child discipline is one of the most important elements of successful parenting. As discipline is defined the process that help children to learn appropriate behaviors and make good choices. Aim: The aim of the present study was to review the literature about the discipline methods. The method οf this study included bibliography research from both the review and the research literature, mainly in the pubmed data base which referred to the discipline methods. Results: In the literature it is ci...

  15. In-house validation study for salmonella unique testing of juice (modification of AOAC official method 2000.07).

    Science.gov (United States)

    Dailianis, Angela E; Hughes, Denise

    2002-01-01

    Following an industry request, a study was undertaken to validate a minor change to the Unique method for testing fruit juice. Twenty foods were tested in the original precollaborative study for TECRA Unique Salmonella test (2000.07). To validate the modification for juice, both the modified method (42 degrees C module incubation with a 5 h replication step) and the current AOAC Method 2000.07 (37 degrees C incubation with a 4 h replication step) were compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM; 8th Ed., 1998) reference method, which uses lactose broth as pre-enrichment medium. Twenty uninoculated replicates, 20 replicates with low-level inoculum (target 1-5 cells/25 g), and 20 replicates with high-level inoculum (target 10-50 cells/25 g) were tested for a single batch of fresh orange juice in accordance with AOAC requirements. There was exact agreement between the 2 Unique methods for all samples and exact agreement between the 2 Unique methods and the BAM method for the uninoculated and high-level inoculum samples. For low-level inoculum, 17 samples were confirmed positive with the new Unique method, 17 with AOAC Method 2000.07, and 14 with the BAM method. PMID:12477197

  16. Maintenance methods

    International Nuclear Information System (INIS)

    The maintenance method applied at the Hague is summarized. The method was developed in order to solve problems relating to: the different specialist fields, the need for homogeneity in the maintenance work, the equipment diversity, the increase of the materials used at the Hague's new facilities. The aim of the method is to create a knowhow formalism, to facilitate maintenance, to ensure the running of the operations and to improve the estimation of the maintenance cost. One of the method's difficulties is the demonstration of the profitability of the maintenance operations

  17. Computing methods

    CERN Document Server

    Berezin, I S

    1965-01-01

    Computing Methods, Volume 2 is a five-chapter text that presents the numerical methods of solving sets of several mathematical equations. This volume includes computation sets of linear algebraic equations, high degree equations and transcendental equations, numerical methods of finding eigenvalues, and approximate methods of solving ordinary differential equations, partial differential equations and integral equations.The book is intended as a text-book for students in mechanical mathematical and physics-mathematical faculties specializing in computer mathematics and persons interested in the

  18. Method 59

    DEFF Research Database (Denmark)

    Kjellsson, G.

    OCDE/GD(94)41. Er supplerende rapport til rapport: The OECD Workshop on Methods for Monitoring Organisms in the Environment, Ottawa 14-17 September 1992. Environment Monograph 90.......OCDE/GD(94)41. Er supplerende rapport til rapport: The OECD Workshop on Methods for Monitoring Organisms in the Environment, Ottawa 14-17 September 1992. Environment Monograph 90....

  19. Ethnographic methods

    OpenAIRE

    G. Gobo

    2011-01-01

    This insightful book explores the most important established and emerging qualitative and quantitative research methods in tourism. The authors provide a detailed overview of the nature of the research method, its use in tourism, the advantages and limitations, and future directions for research.

  20. Numerical Methods

    OpenAIRE

    Dusan Maga

    2004-01-01

    Presented paper is based on authors experience on numerical methods of field solution, mostly magnetic. This paper, as the first one of prepared series, deals with mathematical apparatus and basic physical principles, as well as with possible short-comings or advantages when using the Finite Difference Method (FDM).

  1. A novel culture method for high level production of heterologous protein in Saccharomyces cerevisiae.

    Science.gov (United States)

    Nagashima, T; Yamamoto, Y; Gomi, K; Kitamoto, K; Kumagai, C

    1994-07-01

    A high level production system for heterologous protein by cold culture of yeast transformants at 15 degrees C was developed. The yeast transformants, carrying a plasmid containing cDNA for Aspergillus oryzae alpha-amylase (Taka-amylase A) or human lysozyme synthetic DNA, were cultivated in a selective medium for 1 or 2 days until full growth at 30 degrees C. The yeast cells were harvested by centrifugation from the culture fluid and then were transferred to YPD medium. These inoculated broths were incubated for 2 days at 15 degrees C and then for another 2 days at 30 degrees C. By the cold culture method described above, higher amounts of Taka-amylase A (28.6 mg/liter) and human lysozyme (6.1 mg/liter) were produced by the yeast transformants compared to those by conventional methods. Heterologous protein productions using YEp, YCp, and YIp types of yeast expression vectors with ADH1 or GAPDH promoter by the cold culture method showed effective productivity of about 2-fold compared to those by the conventional method of culture at 30 degrees C. The high level production of heterologous protein by this method was not specific to the S. cerevisiae strains examined. PMID:7765252

  2. A Study on the Antibacterial Activity Of Zno Nanoparticles Prepared By Combustion Method against E Coli

    Directory of Open Access Journals (Sweden)

    Durga Prasad

    2014-06-01

    Full Text Available Crystalline Zinc Oxide (ZnO nanoparticles were synthesized by low temperature solution combustion method using Oxalyl dihydrazide (ODH as fuel, at much lower temperature (300oC. X-ray diffraction (XRD confirmed the formation of wurtzite-structured pure ZnO No peaks from any else phases of ZnO and no impurity peaks were observed, indicating the high purity of the obtained hexagonal ZnO nanocrystals. The antibacterial activity of the formed nano ZnO were investigated against the pathogenic bacteria namely against E-coli. The bacteriological test is performed in Luria-Bertani and Nutrient agar media on solid agar plates and liquid broth system using different concentration of ZnO by standard microbial method. We have used both colony counting method and disk diffusion method. In both the methods ZnO nanoparticles with 100microg/L showed best antibacterial activity, and further studies on destruction of bacterial genomic DNA was done using PCR and gel electrophoresis revealed the DNA fragment bands, this activity might be due to surface charge interactions between the particles and cells. Free radical scavenging properties of the particles might have helped in cell wall disruption, and drastic antimicrobial action.

  3. Comparison of the activities of amphotericin B, itraconazole, and voriconazole against clinical and environmental isolates of Aspergillus species

    Directory of Open Access Journals (Sweden)

    Richa Misra

    2011-01-01

    Full Text Available Background: Invasive fungal infections are a significant cause of morbidity and mortality in immunocompromised populations. Aims: To evaluate the susceptibility pattern of our isolates against amphotericin B, itraconazole, and voriconazole and to compare the antifungal activities of these agents with each other against the Aspergillus species tested. Settings and Design: A prospective study was designed to include clinical and environmental isolates of Aspergillus species. Materials and Methods: 420 sputum samples, 70 bronchoalveolar lavage fluids, 160 oral washings, and 47 environmental samples were collected. Direct microscopy by potassium hydroxide and lactophenol cotton blue mounts followed by culture on Sabourad`s dextrose agar (SDA was done. Susceptibility testing was performed by the broth microdilution technique as per Clinical Laboratory Standards Institute standards (M-38A. Additionally, all the isolates were also tested by the colorimetric microdilution technique using Alamar Blue dye. Statistical Analysis: It was done by the Chi-square test and Z-test using SPSS statistical software version 12.0. Results and Conclusion: Twenty-seven isolates (47.3% were recovered from patients with chronic bronchial asthma followed by fibrocavitary pulmonary tuberculosis in 9 (15.7%, allergic bronchopulmonary aspergillosis (ABPA in 6 cases (10.5%, bronchiectasis in 3 (5.2%, bronchogenic carcinoma in 5 (8.7% and those receiving radiotherapy for head and neck cancer 7 (12.2%. Thirteen environmental isolates were also included in the study. The most common isolate was A. fumigatus 28 (40%, followed by A. niger 22 (31%, A. flavus 13 (19%, and A. terreus 7(10%. All isolates were susceptible to amphotericin B, itraconazole, and voriconazole. Among the three agents tested, voriconazole exhibited lowest MICs (≤1 μg/ml against all Aspergillus species.

  4. Dosimetry methods

    DEFF Research Database (Denmark)

    McLaughlin, W.L.; Miller, A.; Kovacs, A.; Mehta, K. K.

    Chemical and physical radiation dosimetry methods, used for the measurement of absorbed dose mainly during the practical use of ionizing radiation, are discussed with respect to their characteristics and fields of application.......Chemical and physical radiation dosimetry methods, used for the measurement of absorbed dose mainly during the practical use of ionizing radiation, are discussed with respect to their characteristics and fields of application....

  5. Ensemble Methods

    Science.gov (United States)

    Re, Matteo; Valentini, Giorgio

    2012-03-01

    Ensemble methods are statistical and computational learning procedures reminiscent of the human social learning behavior of seeking several opinions before making any crucial decision. The idea of combining the opinions of different "experts" to obtain an overall “ensemble” decision is rooted in our culture at least from the classical age of ancient Greece, and it has been formalized during the Enlightenment with the Condorcet Jury Theorem[45]), which proved that the judgment of a committee is superior to those of individuals, provided the individuals have reasonable competence. Ensembles are sets of learning machines that combine in some way their decisions, or their learning algorithms, or different views of data, or other specific characteristics to obtain more reliable and more accurate predictions in supervised and unsupervised learning problems [48,116]. A simple example is represented by the majority vote ensemble, by which the decisions of different learning machines are combined, and the class that receives the majority of “votes” (i.e., the class predicted by the majority of the learning machines) is the class predicted by the overall ensemble [158]. In the literature, a plethora of terms other than ensembles has been used, such as fusion, combination, aggregation, and committee, to indicate sets of learning machines that work together to solve a machine learning problem [19,40,56,66,99,108,123], but in this chapter we maintain the term ensemble in its widest meaning, in order to include the whole range of combination methods. Nowadays, ensemble methods represent one of the main current research lines in machine learning [48,116], and the interest of the research community on ensemble methods is witnessed by conferences and workshops specifically devoted to ensembles, first of all the multiple classifier systems (MCS) conference organized by Roli, Kittler, Windeatt, and other researchers of this area [14,62,85,149,173]. Several theories have been

  6. Ensemble Methods

    Science.gov (United States)

    Re, Matteo; Valentini, Giorgio

    2012-03-01

    Ensemble methods are statistical and computational learning procedures reminiscent of the human social learning behavior of seeking several opinions before making any crucial decision. The idea of combining the opinions of different "experts" to obtain an overall “ensemble” decision is rooted in our culture at least from the classical age of ancient Greece, and it has been formalized during the Enlightenment with the Condorcet Jury Theorem[45]), which proved that the judgment of a committee is superior to those of individuals, provided the individuals have reasonable competence. Ensembles are sets of learning machines that combine in some way their decisions, or their learning algorithms, or different views of data, or other specific characteristics to obtain more reliable and more accurate predictions in supervised and unsupervised learning problems [48,116]. A simple example is represented by the majority vote ensemble, by which the decisions of different learning machines are combined, and the class that receives the majority of “votes” (i.e., the class predicted by the majority of the learning machines) is the class predicted by the overall ensemble [158]. In the literature, a plethora of terms other than ensembles has been used, such as fusion, combination, aggregation, and committee, to indicate sets of learning machines that work together to solve a machine learning problem [19,40,56,66,99,108,123], but in this chapter we maintain the term ensemble in its widest meaning, in order to include the whole range of combination methods. Nowadays, ensemble methods represent one of the main current research lines in machine learning [48,116], and the interest of the research community on ensemble methods is witnessed by conferences and workshops specifically devoted to ensembles, first of all the multiple classifier systems (MCS) conference organized by Roli, Kittler, Windeatt, and other researchers of this area [14,62,85,149,173]. Several theories have been

  7. Discipline methods

    Directory of Open Access Journals (Sweden)

    Maria Kikila

    2012-01-01

    Full Text Available Child discipline is one of the most important elements of successful parenting. As discipline is defined the process that help children to learn appropriate behaviors and make good choices. Aim: The aim of the present study was to review the literature about the discipline methods. The method οf this study included bibliography research from both the review and the research literature, mainly in the pubmed data base which referred to the discipline methods. Results: In the literature it is cited that discipline is not the same as punishment because is more related to teaching. The ultimate goal of effective punishment is to help the child in exercising self-control, accountability and respect. Children raised without proper discipline, are at risk for a variety of behavioral and emotional problems. Over the last decades, many child discipline techniques have been suggested that vary according to the age of the child, the parenting style and the severity of the behavior. The most effective methods are giving rewards and guidance to make good choices, explaining negative consequences for disobedience without issuing threats, teaching of moral behaviors by displaying parents' good and moral behaviors. Another commonly accepted discipline method is "the consequences' which permits children to have the dignity of dealing with results of their behavior. Conclusions: Discipline consists a tool for parents for the reason that it trains children to think their improper behavior and the following consequences as well as the possible solutions to each problem.

  8. Statistical methods

    CERN Document Server

    Szulc, Stefan

    1965-01-01

    Statistical Methods provides a discussion of the principles of the organization and technique of research, with emphasis on its application to the problems in social statistics. This book discusses branch statistics, which aims to develop practical ways of collecting and processing numerical data and to adapt general statistical methods to the objectives in a given field.Organized into five parts encompassing 22 chapters, this book begins with an overview of how to organize the collection of such information on individual units, primarily as accomplished by government agencies. This text then

  9. Statistical methods

    CERN Document Server

    Freund, Rudolf J; Wilson, William J

    2010-01-01

    Statistical Methods, 3e provides students with a working introduction to statistical methods offering a wide range of applications that emphasize the quantitative skills useful across many academic disciplines. This text takes a classic approach emphasizing concepts and techniques for working out problems and intepreting results. The book includes research projects, real-world case studies, numerous examples and data exercises organized by level of difficulty. This text requires that a student be familiar with algebra. New to this edition: NEW expansion of exercises a

  10. Digital Methods

    NARCIS (Netherlands)

    R. Rogers

    2013-01-01

    In Digital Methods, Richard Rogers proposes a methodological outlook for social and cultural scholarly research on the Web that seeks to move Internet research beyond the study of online culture. It is not a toolkit for Internet research, or operating instructions for a software package; it deals wi

  11. Characterization methods

    Energy Technology Data Exchange (ETDEWEB)

    Glass, J.T. [North Carolina State Univ., Raleigh (United States)

    1993-01-01

    Methods discussed in this compilation of notes and diagrams are Raman spectroscopy, scanning electron microscopy, transmission electron microscopy, and other surface analysis techniques (auger electron spectroscopy, x-ray photoelectron spectroscopy, electron energy loss spectroscopy, and scanning tunnelling microscopy). A comparative evaluation of different techniques is performed. In-vacuo and in-situ analyses are described.

  12. Critical assessment of suitable methods used for determination of antibacterial properties at photocatalytic surfaces

    Energy Technology Data Exchange (ETDEWEB)

    Krysa, Josef, E-mail: Josef.Krysa@vscht.cz [Institute of Chemical Technology Prague, Department of Water Technology and Environmental Engineering, Technicka 5, CZ-166 28 Prague (Czech Republic); Musilova, Eva [Institute of Chemical Technology Prague, Department of Water Technology and Environmental Engineering, Technicka 5, CZ-166 28 Prague (Czech Republic); Zita, Jiri [Institute of Chemical Technology Prague, Department of Inorganic Technology, Technicka 5, CZ-166 28 Prague (Czech Republic)

    2011-11-15

    Highlights: {yields} Methods for antibacterial activity of irradiated TiO{sub 2} films were critically assessed. {yields} As test bacteria gram negative E. coli and gram positive E. faecalis were employed. {yields} The ISO glass adhesion method is more appropriate than method with bacteria suspension. {yields} Some improvements of the ISO method were suggested. - Abstract: This work describes the development of methods necessary for antibacterial effect evaluation on irradiated TiO{sub 2} layers. Two methods using bacteria suspensions and the glass adhesion method (based on ISO 27447:2009(E)) were critically assessed and compared. As test bacteria gram negative Escherichia coli and gram positive Enterococcus faecalis were employed. The method using 50 cm{sup 3} of bacteria suspension is convenient for testing layers with strong antibacterial effect (prepared from powder photocatalysts). For the evaluation of the antibacterial effect of sol gel layers, the glass adhesion method based on the ISO is more appropriate than the method with 3 cm{sup 3} of bacteria suspension. The reason is that the later does not allow a distinction between the inhibition effect of TiO{sub 2} and UV light itself. Some improvements of the ISO method were suggested, namely the use of gelatinous pills (CCM) of bacteria, using saline solution instead of nutrient broth for bacteria suspension preparation and the application of selective media for bacteria cultivation. Decreasing the light intensity from 0.6 mW cm{sup -2} to 0.2 mW cm{sup -2} (fulfilling the requirements of the ISO) results in almost negligible effect of UV light itself, thus enabling proper testing of the antibacterial properties of TiO{sub 2} thin films.

  13. Numerical methods

    CERN Document Server

    Dahlquist, Germund

    2003-01-01

    ""Substantial, detailed and rigorous . . . readers for whom the book is intended are admirably served."" - MathSciNet (Mathematical Reviews on the Web), American Mathematical Society.Practical text strikes fine balance between students' requirements for theoretical treatment and needs of practitioners, with best methods for large- and small-scale computing. Prerequisites are minimal (calculus, linear algebra, and preferably some acquaintance with computer programming). Text includes many worked examples, problems, and an extensive bibliography.

  14. Protein expression on Cr resistant microorganism using electrophoresis method

    Directory of Open Access Journals (Sweden)

    SAJIDAN

    2009-01-01

    Full Text Available Fatmawati U, Suranto, Sajidan. 2009. Protein expression on Cr resistant microorganism using electrophoresis method. Nusantara Bioscience 1: 31-37. Hexavalent chromium (Cr(VI is known as toxic heavy metals, so the need is reduced to Cr(III is much less toxicity. Pseudomonas aeruginosa, Pseudomonas putida, Klebsiella pneumoniae, Pantoea sp. and Saccharomyces cerevisiae are resistant Cr(VI microorganism and have ability to reduce Cr(VI. The aim of this research is to know ability of microorganism to reduce Cr(VI and to know protein band pattern between Cr(VI resistant microorganism and non resistant microorganism which inoculated on LB broth. SDS-PAGE was used to indentify protein expression. While, Cr(VI concentration was identified by 1.5 diphenylcarbazide method. The quantitative data was analyzed by two factorial ANOVA that continued with DMRT at 1% level test. The qualitative data i.e. protein expression analyzed by relative mobility (Rf. The results showed that the ability of microorganisms to reduce Cr(VI at initial concentration of 0.5 ppm, 1 ppm, 5 ppm and 10 ppm may vary, the average percentage of the ability of each microorganism in reducing Cr(VI is P. putida (65% > S. cerevisiae (64.45% >. P. aeruginosa (60.73% > Pantoea sp. (50.22% > K. pneumoniae (47.82% > without microorganisms (34.25%. The adding microorganisms have significantly influenced toward reduction of Cr(VI. The SDS-PAGE shows that protein expression between resistant and not resistant microorganisms are no different, but resistant microorganisms have more protein (protein band is thicker.

  15. Antimicrobial constituents of the leaves of Mikania micrantha H. B. K.

    Directory of Open Access Journals (Sweden)

    Yan Li

    Full Text Available BACKGROUND: To isolate plant-derived compounds with antimicrobial activity from the leaves of Mikania micrantha, to determine the compounds configuration, and to evaluate their antimicrobial activity against eight plant pathogenic fungi (Exserohilum turcicum, Colletotrichum lagenarium, Pseudoperonispora cubensis, Botrytis cirerea, Rhizoctonia solani, Phytophthora parasitica, Fusarium solani, and Pythium aphanidermatum, and four plant pathogenic bacteria (gram negative bacteria: Ralstonia dolaanacearum, Xanthomonas oryzae pv. Oryzae, Xanthomonas Campestris pv. Vesicatoria, and Xanthomonas campestris pv. Citri, and four bacteria (gram positive bacteria: Staphyloccocus aureus, Bacillus subtilis, Micrococcus luteus, and Bacillus cereus. METHODS AND RESULTS: Antimicrobial constituents of the leaves of M. micrantha were isolated using bioactivity- guided fractionation. The antifungal activity of the isolated compounds was evaluated by the inhibit hypha growth method and inhibit spore germination method. Characterization of antibacterial activity was carried out using the minimum inhibitory concentrations (MICs and the minimum bactericidal concentrations (MBCs. MIC and MBC were determined by the broth microdilution method. Six compounds - deoxymikanolide, scandenolide, dihydroscandenolide, mikanolide, dihydromikanolide, and m - methoxy benzoic acid - have been isolated from leaves of Mikania micrantha H. B. K. Deoxymikanolide, scandenolide, and dihydroscandenolide were new compounds. The result of bioassay showed that all of isolated compounds were effective against tested strains and deoxymikanolide showed the strongest activity. CONCLUSIONS AND SIGNIFICANCE: The leaves of M. micrantha may be a promising source in the search for new antimicrobial drugs due to its efficacy and the broadest range. Meanwhile, adverse impact of M. micrantha will be eliminated.

  16. Bioactive proifle of Plakortis nigra, a sea sponge from Mauritius Islands

    Institute of Scientific and Technical Information of China (English)

    Sandrine Tangman; Joyce Govinden Soulange; Daniel Marie

    2015-01-01

    Objective:To investigate the in vitro antibacterial and antioxidant activities of crude and fractionated extracts of the Plakortis nigra (P. nigra) sea sponge from Mauritius sea waters. Methods:Preliminary qualitative chemical screening of the sponge extracts was conducted by using standard methods while the total phenolic content (TPC) was estimated through the Folin-Ciocalteu method. Antibacterial activity was evaluated against Staphylococcus aureus, Escherichia coli and Proteus mirabilis. The minimum inhibitory concentration was determined by the broth microdilution method. All sponge extracts were assessed for antioxidant activity via the 1,1-diphenyl-2-picrylhydrazyl free radical scavenging in vitro model. Results:Alkaloids, phenols, steroids, terpenoids, tannins and saponins were detected in the sponge extracts and TPC varied from (2.280±0.072) mg to (12.790±0.236) mg gallic acid equivalents per gram extract (P Conclusions:The present study validates the antioxidant and antibacterial activity of marine sponge (P. nigra) extracts and depicts the sea sponge as a potential source of pharmaceutical leads against infectious and degenerative diseases.

  17. In Vitro Activities of Ertapenem (MK-0826) against Recent Clinical Bacteria Collected in Europe and Australia

    Science.gov (United States)

    Livermore, David M.; Carter, Michael W.; Bagel, Simone; Wiedemann, Bernd; Baquero, Fernando; Loza, Elena; Endtz, Hubert P.; van den Braak, Nicole; Fernandes, Clarence J.; Fernandes, Lorna; Frimodt-Moller, Niels; Rasmussen, Laura S.; Giamarellou, Helen; Giamarellos-Bourboulis, Evangelos; Jarlier, Vincent; Nguyen, Jacqueline; Nord, Carl-Erik; Struelens, Marc J.; Nonhoff, Caire; Turnidge, John; Bell, Jan; Zbinden, Reinhard; Pfister, Stefan; Mixson, Lori; Shungu, Daniel L.

    2001-01-01

    those of the broth microdilution method for ertapenem. PMID:11353638

  18. In vitro study on the antimicrobial effect of hydroalcoholic extracts from Mentha arvensis L. (Lamiaceae against oral pathogens=Estudo in vitro do efeito antimicrobiano dos extratos hidroalcólicos de Mentha arvensis L. (Lamiaceae contra patógenos orais

    Directory of Open Access Journals (Sweden)

    Rogério Antonio Freitag

    2012-10-01

    Full Text Available In vitro tests could be a valuable tool for the evaluation of medicinal plants’ antimicrobial activity. Mentha arvensis of the Lamiaceae family is one of the most frequently traditional plants used in Brazil. Hydroalcoholic extracts of M. arvensis were analyzed for antimicrobial activity on Streptococcus mutans, Streptococcus sobrinus and Candida albicans. Three different assays (agar diffusion, broth macro- and micro-dilution methods were used to evaluate antimicrobial activity. Although hydroalcoholic extracts of M. arvensis did not show any antibacterial effect, its antifungal activity against C. albicans was revealed. According to the micro-dilution broth assay, MIC of the hydroalcoholic extract from leaves of M. arvensis on Candida albicans strains ranged between 625 and 2500 g mL-1. Results suggest that M. arvensis hydroalcoholic extract may be considered a potentially antifungal agent against C. albicans, and a possible item for human antibiotic therapy. However, further biological tests on the plant’s efficacy and side-effects are necessary before its use on humans. Testes in vitro podem ser uma ferramenta valiosa para a avaliação da atividade antimicrobiana de plantas medicinais. Mentha arvensis é uma das plantas medicinais brasileiras mais frequentemente utilizadas e pertence à família Lamiaceae. No presente estudo, extratos hidroalcólicos de M. arvensis foram analisados quanto à sua atividade antimicrobiana sobre Streptococcus mutans, Streptococcus sobrinus e Candida albicans. Três diferentes ensaios (métodos de difusão em ágar, macro e microdiluição em caldo foram utilizados para avaliação da atividade antimicrobiana. Embora os extratos hidroalcólicos de M. arvensis não demonstraram qualquer efeito antibacteriano, eles apresentaram atividade antifúngica contra C. albicans. Baseado no ensaio de microdiluição em caldo, a CIM do extrato hidroalcólico das folhas de M. arvensis sobre cepas de C. albicans

  19. A comparative study of cultural methods for the detection of Salmonella in feed and feed ingredients

    Directory of Open Access Journals (Sweden)

    Haggblom Per

    2009-02-01

    Full Text Available Abstract Background Animal feed as a source of infection to food producing animals is much debated. In order to increase our present knowledge about possible feed transmission it is important to know that the present isolation methods for Salmonella are reliable also for feed materials. In a comparative study the ability of the standard method used for isolation of Salmonella in feed in the Nordic countries, the NMKL71 method (Nordic Committee on Food Analysis was compared to the Modified Semisolid Rappaport Vassiliadis method (MSRV and the international standard method (EN ISO 6579:2002. Five different feed materials were investigated, namely wheat grain, soybean meal, rape seed meal, palm kernel meal, pellets of pig feed and also scrapings from a feed mill elevator. Four different levels of the Salmonella serotypes S. Typhimurium, S. Cubana and S. Yoruba were added to each feed material, respectively. For all methods pre-enrichment in Buffered Peptone Water (BPW were carried out followed by enrichments in the different selective media and finally plating on selective agar media. Results The results obtained with all three methods showed no differences in detection levels, with an accuracy and sensitivity of 65% and 56%, respectively. However, Müller-Kauffmann tetrathionate-novobiocin broth (MKTTn, performed less well due to many false-negative results on Brilliant Green agar (BGA plates. Compared to other feed materials palm kernel meal showed a higher detection level with all serotypes and methods tested. Conclusion The results of this study showed that the accuracy, sensitivity and specificity of the investigated cultural methods were equivalent. However, the detection levels for different feed and feed ingredients varied considerably.

  20. Antimicrobial action of zinc oxide nanoparticles in combination with ciprofloxacin and ceftazidime against multidrug-resistant Acinetobacter baumannii.

    Science.gov (United States)

    Ghasemi, F; Jalal, R

    2016-09-01

    Acinetobacter baumannii is a serious concern amongst hospitalised patients worldwide and its resistance to antibiotics has emerged as a threat to public health in recent years. Metal oxide nanoparticles were found to be effective for overcoming bacterial resistance owing to their antibacterial activities. The aim of this study was to investigate the combined effects of zinc oxide nanoparticles (ZnO-NPs) and the conventional antibiotics ciprofloxacin and ceftazidime as well as their mechanisms of action against resistant A. baumannii. ZnO-NPs were prepared by the solvothermal method and were characterised by various methods. Broth microdilution and disk diffusion methods were used to determine the antibacterial activities of ciprofloxacin and ceftazidime antibiotics in the absence and presence of a subinhibitory concentration of ZnO-NPs. The mechanism of action of ZnO-NPs alone and in combination with these antibiotics was assessed by flow cytometry, DNA extraction, fluorescence and scanning electron microscopy. The results showed that the antibacterial activities of both antibiotics increased in the presence of a subinhibitory concentration of ZnO-NPs. Combination of ZnO-NPs with antibiotics increased the uptake of antibiotics and changed the bacterial cells from rod to cocci forms. Bacterial filamentation was also observed and exhibited no DNA fragmentation. In conclusion, the results of this study indicate that ZnO-NPs potentiate the antimicrobial action of ciprofloxacin and ceftazidime. A mechanism is proposed to explain this phenomenon. PMID:27530853

  1. EVALUATION OF ANTIBACTERIAL ACTIVITY OF CAFFEINE

    Directory of Open Access Journals (Sweden)

    Pawar Pruthviraj

    2011-04-01

    Full Text Available The present study was carried out with water soluble portion and pure solvent of the acetone, ethanol, methanol, acetonitrile, water extracts of leaves and leaf buds of Camellia sinensis (green tea, and beans of Coffea arabica (coffee. Caffeine (3,7-dihydro-1, 3,7-trimethyl-1H-purine-2,6-dione was isolated from both plants using a liquid-liquid extraction method, detected on thin layer chromatography (TLC plates in comparison with standard caffeine, which served as a positive control. After performing the gross behavioral study, the Antibacterial activity was evaluated against Gram-negative bacteria included; Escherichia coli, Proteus mirabilis, Klebsiella pneumonia and Pseudomonas aeruginosa Both compounds at a concentration of 2 mg/ml showed similar antibacterial activities against all tested bacteria, except for P. mirabilis, and the highest inhibitory effect was observed against P. aeruginosa using a modified agar diffusion method. The minimal inhibitory concentration (MIC of caffeine was determined using a broth microdilution method in 96 multi-well microtitre plates. MIC values ranged from 65.5 to 250.0 µg/ml for the caffeine isolated from coffee and 65.5 to 500.0 µg/ml for green tea caffeine. Combination results showed additive effects against most pathogenic bacteria especially for P. aeruginosa, using both antibacterial assays.

  2. Influence of different ions doping on the antibacterial properties of MgO nanopowders

    International Nuclear Information System (INIS)

    Compared with other inorganic antibacterial agents, magnesium oxide (MgO) nanopowders exhibit a unique antibacterial mechanism and various advantages in applications, having attracted extensive attention. In this study, MgO nanopowders doped with different ions (Li+, Zn2+ and Ti4+) were synthesized by a sol–gel method, respectively. The structures and morphologies of the as-obtained precursors and nanopowders were characterized and confirmed by X-ray diffraction (XRD), transmission electron microscope (TEM) and X-ray photoelectron spectroscopy (XPS) analysis. The influence of three metal ions doping on the antibacterial properties of MgO nanopowders was also investigated by their bactericidal activity against Escherichia coli (E. coli, ATCC 25922) using the broth microdilution method and the agar method. The results show that Li-doped MgO exhibits better antibacterial activity, Zn-doped and Ti-doped MgO display poorer antibacterial activity than pure MgO. It can be concluded that the influence of different ions doping on the antibacterial properties of MgO mainly lies on oxygen vacancies and basicity of nanopowders.

  3. Susceptibility of Staphylococcus aureus Clinical Isolates to Propolis Extract Alone or in Combination with Antimicrobial Drugs

    Directory of Open Access Journals (Sweden)

    Mieczysław Sajewicz

    2013-08-01

    Full Text Available The objective of this study was to assess in vitro the antimicrobial activity of ethanolic extract of Polish propolis (EEPP against methicillin-sensitive Staphylococcus aureus (MSSA and methicillin-resistant Staphylococcus aureus (MRSA clinical isolates. The combined effect of EEPP and 10 selected antistaphylococcal drugs on S. aureus clinical cultures was also investigated. EEPP composition was analyzed by a High Performance Liquid Chromatography (HPLC method. The flavonoid compounds identified in Polish Propolis included flavones, flavonones, flavonolols, flavonols and phenolic acids. EEPP displayed varying effectiveness against twelve S. aureus strains, with minimal inhibitory concentration (MIC within the range from 0.39 to 0.78 mg/mL, determined by broth microdilution method. The average MIC was 0.54 ± 0.22 mg/mL, while calculated MIC50 and MIC90 were 0.39 mg/mL and 0.78 mg/mL, respectively. The minimum bactericidal concentration (MBC of the EEPP ranged from 0.78 to 3.13 mg/mL. The in vitro combined effect of EEPP and 10 antibacterial drugs was investigated using disk diffusion method-based assay. Addition of EEPP to cefoxitin (FOX, clindamycin (DA, tetracycline (TE, tobramycin (TOB, linezolid (LIN, trimethoprim+sulfamethoxazole (SXT, penicillin (P, erythromycin (E regimen, yielded stronger, cumulative antimicrobial effect, against all tested S. aureus strains than EEPP and chemotherapeutics alone. In the case of ciprofloxacin (CIP and chloramphenicol (C no synergism with EEPP was observed.

  4. In vitro SCREENING ANTIBACTERIAL ACTIVITY OF Bidens pilosa LINNÉ AND Annona crassiflora MART. AGAINST OXACILLIN RESISTANT Staphylococcus aureus (ORSA FROM THE AERIAL ENVIRONMENT AT THE DENTAL CLINIC

    Directory of Open Access Journals (Sweden)

    Jeferson Junior da Silva

    2014-07-01

    Full Text Available Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves and Annona crassiflora (rind fruit, stem, leaves, seed and pulp were obtained with ethanol: water (7:3, v/v and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05. Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90 for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.

  5. In vitro Synergistic Antimicrobial Activity of Romanian Propolis and Antibiotics against Escherichia coli Isolated from Bovine Mastitis

    Directory of Open Access Journals (Sweden)

    Mihaela NICULAE

    2015-12-01

    Full Text Available The study was aimed to characterize the chemical composition and the antimicrobial activity of Romanian propolis ethanolic extracts (EEP against antibiotic-sensitive and antibiotic-resistant E. coli strains isolated from bovine mastitis. The preliminary antimicrobial screening was performed by a disk diffusion method, followed by determination of minimum inhibitory concentrations (MIC and minimum bactericidal concentrations (MBC based on broth microdilution assay; further, the synergistic action of propolis with antimicrobial drugs was assessed by a disk diffusion method on agar containing subinhibitory concentrations of propolis. For the chemical characterisation of EEP, the flavonoids (flavones/flavonols, flavanones/dihydroflavonols and total phenolics were evaluated by spectrophotometric methods. The phenolic compounds of these extracts were also determined using HPLC. The results indicated for Romanian propolis ethanolic extracts the typical poplar composition profile with flavonoids and phenolic acids as main biological active compounds, with chromatographic analysis data confirmed also spectrophotometrically. In addition, positively correlated with the chemical composition, a strong antimicrobial efficacy was exhibited towards E. coli strains, along with interesting synergistic interaction with antibiotics that can be further investigated to obtain propolis-based formulation with antibacterial properties. Subsequent in vitro and in vivo studies evaluating the safety and efficacy are intended to consider propolis in veterinary therapeutic protocols.

  6. WELDING METHOD

    Science.gov (United States)

    Cornell, A.A.; Dunbar, J.V.; Ruffner, J.H.

    1959-09-29

    A semi-automatic method is described for the weld joining of pipes and fittings which utilizes the inert gasshielded consumable electrode electric arc welding technique, comprising laying down the root pass at a first peripheral velocity and thereafter laying down the filler passes over the root pass necessary to complete the weld by revolving the pipes and fittings at a second peripheral velocity different from the first peripheral velocity, maintaining the welding head in a fixed position as to the specific direction of revolution, while the longitudinal axis of the welding head is disposed angularly in the direction of revolution at amounts between twenty minutas and about four degrees from the first position.

  7. Radiochemical methods

    International Nuclear Information System (INIS)

    This little volume is one of an extended series of basic textbooks on analytical chemistry produced by the Analytical Chemistry by Open Learning project in the UK. Prefatory sections explain its mission, and how to use the Open Learning format. Seventeen specific sections organized into five chaptrs begin with a general discussion of nuclear properties, types, and laws of nuclear decay and proceeds to specific discussions of three published papers (reproduced in their entirety) giving examples of radiochemical methods which were discussed in the previous chapter. Each section begins with an overview, contains one or more practical problems (called self-assessment questions or SAQ's), and concludes with a summary and a list of objectives for the student. Following the main body are answers to the SAQ's, and several tables of physical constants, SI prefixes, etc. A periodic table graces the inside back cover

  8. Solidification method

    International Nuclear Information System (INIS)

    Purpose: To subject the condensate of steam discharged from the turbine of a BWR type power plant to ion-exchange processing, and thereafter to render sodium sulfate containing radioactive liquid wastes produced when the ion-exchange resin thus obtained are reproduced into asphalt solidified bodies stable even in water. Method: In the above described liquid wastes, there is added a water-soluble calcium compound such as calcium nitrate or the like. In this case, when the quantity of the water-soluble calcium compound added is 7 to 8% with respect to sulfuric acid ions on the basis of the weight of calcium ions, swelling and porosity of solidified bodies can be prevented. (Kamimura, M.)

  9. IMPLEMENTATION METHOD

    Directory of Open Access Journals (Sweden)

    Cătălin LUPU

    2009-06-01

    Full Text Available In this article presents applications of “Divide et impera” method using object -oriented programming in C #.Main advantage of using the "divide et impera" cost in that it allows software to reduce the complexity of the problem,sub-problems that were being decomposed and simpler data sharing in smaller groups of data (eg sub -algorithmQuickSort. Object-oriented programming means programs with new types that integrates both data and methodsassociated with the creation, processing and destruction of such data. To gain advantages through abstractionprogramming (the program is no longer a succession of processing, but a set of objects to life, have differentproperties, are capable of specific action s and interact in the program. Spoke on instantiation new techniques,derivation and polimorfismul object types.

  10. Comparison of growth methods and biological activities of brazilian marine Streptomyces

    Directory of Open Access Journals (Sweden)

    A. C. Granato

    2013-03-01

    Full Text Available The present work describes the study of the growth and the cytotoxic and antitumor activities of the extracts of the marine microorganisms Streptomyces acrymicini and Streptomyces cebimarensis, the latter a new strain. Both microorganisms were collected from coastal marine sediments of the north coast of São Paulo state. Growth was performed in a shaker and in a bioreactor using Gym medium and the broths of both microorganisms were extracted with ethyl acetate and n-butanol. Three extracts, two organic and one aqueous, from each microorganism were obtained and tested for cytotoxic and antitumor activity using the SF-295 (Central Nervous System, HCT-8 (Colon cell lines, and the MTT (3-(4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide method. The growth methods were compared and show that, although the shaker presented reasonable results, the bioreactor represents the best choice for growth of these microorganisms. The biological activity of the different extracts was evaluated and it was demonstrated that the growth methodology may influence the secondary metabolite production and the biological activity.

  11. Methods used in preclinical assessment of anti-Buruli ulcer agents: A global perspective.

    Science.gov (United States)

    Tsouh, Patrick Valere Fokou; Addo, Phyllis; Yeboah-Manu, Dorothy; Boyom, Fabrice Fekam

    2015-01-01

    Buruli ulcer (BU) caused by Mycobacterium ulcerans is the third most common chronic mycobacterial infection in humans. Approximately 5000 cases are reported annually from at least 33 countries around the globe, especially in rural African communities. Even though anti-mycobacterial therapy is often effective for early nodular or ulcerative lesions, surgery is sometimes employed for aiding wound healing and correction of deformities. The usefulness of the antibiotherapy nonetheless is challenged by huge restrictive factors such as high cost, surgical scars and loss of income due to loss of man-hours, and in some instances employment. For these reasons, more effective and safer drugs are urgently needed, and research programs into alternative therapeutics including investigation of natural products should be encouraged. There is the need for appropriate susceptibility testing methods for the evaluation of potency. A number of biological assay methodologies are in current use, ranging from the classical agar and broth dilution assay formats, to radiorespirometric, dye-based, and fluorescent/luminescence reporter assays. Mice, rats, armadillo, guinea pigs, monkeys, grass cutters and lizards have been suggested as animal models for Buruli ulcer. This review presents an overview of in vitro and in vivo susceptibility testing methods developed so far for the determination of anti-Buruli ulcer activity of natural products and derivatives. PMID:25792087

  12. Novel Plasmid Transformation Method Mediated by Chrysotile, Sliding Friction, and Elastic Body Exposure

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    Naoto Yoshida

    2007-01-01

    Full Text Available Escherichia coli as a plasmid recipient cell was dispersed in a chrysotile colloidal solution, containing chrysotile adsorbed to plasmid DNA (chrysotile-plasmid cell mixture. Following this, the chrysotile-plasmid cell mixture was dropped onto the surface of an elastic body, such as agarose, and treated physically by sliding a polystyrene streak bar over the elastic body to create friction. Plasmid DNA was easily incorporated into E. coli, and antibiotic resistance was conferred by transformation. The transformation efficiency of E. coli cultured in solid medium was greater than that of E. coli cultured in broth. To obtain greater transformation efficiency, we attempted to determine optimal transformation conditions. The following conditions resulted in the greatest transformation efficiency: the recipient cell concentration within the chrysotileplasmid cell mixture had an optical density greater than or equal to 2 at 550 nm, the vertical reaction force applied to the streak bar was greater than or equal to 40 g, and the rotation speed of the elastic body was greater than or equal to 34 rpm. Under these conditions, we observed a transformation efficiency of 107 per μg plasmid DNA. The advantage of achieving bacterial transformation using the elastic body exposure method is that competent cell preparation of the recipient cell is not required. In addition to E. coli, other Gram negative bacteria are able to acquire plasmid DNA using the elastic body exposure method.

  13. Decontaminating method

    International Nuclear Information System (INIS)

    Purpose: To enable to decontaminate an object having a complicate surface and also remove hard cruds by a simple device. Method: An object to be decontaminated is placed in a water vessel and steams jetted out from a steam nozzle are condensated at the surface of the object to be decontaminated and decontamination is conducted by impact shocks upon elimination of bubbles. The inside of the water vessel is filled with liquid and steams jetted out from the steam nozzle are cooled by the liquid and condensated. The steams are supplied with a steam supply device by way of steam supply pipeways to the steam nozzle. Since the temperature of the liquid in the water vessel is elevated upon condensation of the jetted steams, the liquids are cooled by the cooling device. Further, since the steams condensated at the surface of the decontaminated object forms water of condensation to increase the water level in the water vessel, the water corresponding to the elevated level is discharged from an overflow pipe. (Kawakami, Y.)

  14. Decontaminating method

    Energy Technology Data Exchange (ETDEWEB)

    Sakurai, Mikio; Hayashi, Tsutomu; Izumi, Masayuki; Sasaki, Hiroshi; Sato, Toru; Owada, Kazuo.

    1987-11-21

    Purpose: To enable to decontaminate an object having a complicate surface and also remove hard cruds by a simple device. Method: An object to be decontaminated is placed in a water vessel and steams jetted out from a steam nozzle are condensated at the surface of the object to be decontaminated and decontamination is conducted by impact shocks upon elimination of bubbles. The inside of the water vessel is filled with liquid and steams jetted out from the steam nozzle are cooled by the liquid and condensated. The steams are supplied with a steam supply device by way of steam supply pipeways to the steam nozzle. Since the temperature of the liquid in the water vessel is elevated upon condensation of the jetted steams, the liquids are cooled by the cooling device. Further, since the steams condensated at the surface of the decontaminated object forms water of condensation to increase the water level in the water vessel, the water corresponding to the elevated level is discharged from an overflow pipe. (Kawakami, Y.).

  15. Combination Therapy of Sophoraflavanone B against MRSA: In Vitro Synergy Testing

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    Su-Hyun Mun

    2013-01-01

    Full Text Available Sophoraflavanone B (SPF-B, a known prenylated flavonoid, was isolated from the roots of Desmodium caudatum. The aim of this study was to determine the antimicrobial synergism of SPF-B combined with antibiotics against methicillin-resistant Staphylococcus aureus (MRSA. MRSA, a multidrug-resistant pathogen, causes both hospital- and community-acquired infections worldwide. The antimicrobial activity of SPF-B was assessed by the broth microdilution method, checkerboard dilution test, and time-kill curve assay. The MIC of SPF-B for 7 strains of S. aureus ranges from 15.6 to 31.25 μg/mL determined. In the checkerboard method, the combinations of SPF-B with antibiotics had a synergistic effect; SPF-B markedly reduced the MICs of the β-lactam antibiotics: ampicillin (AMP and oxacillin (OXI; aminoglycosides gentamicin (GET; quinolones ciprofloxacin (CIP and norfloxacin (NOR against MRSA. The time-kill curves assay showed that a combined SPF-B and selected antibiotics treatment reduced the bacterial counts below the lowest detectable limit after 24 h. These data suggest that the antibacterial activity of SPF-B against MRSA can be effectively increased through its combination with three groups of antibiotics (β-lactams, aminoglycosides, and quinolones. Our research can be a valuable and significant source for the development of a new antibacterial drug with low MRSA resistance.

  16. Antimicrobial activity of Gel-entrapped catechins toward oral microorganisms.

    Science.gov (United States)

    Tamura, Muneaki; Saito, Hideo; Kikuchi, Kuniyoshi; Ishigami, Tomohiko; Toyama, Yoshio; Takami, Masao; Ochiai, Kuniyasu

    2011-01-01

    The oral cavity contains almost half of the commensal bacterial population present in the human body. An increase in the number of these microorganisms may result in systemic diseases such as infective endocarditis and aspiration pneumonia as well as oral infections. It is essential to control the total numbers of these microorganisms in order to suppress disease onset. Thus, we examined the antimicrobial activity of a newly developed gel-entrapped catechin (GEC) preparation against oral microorganisms. The minimum inhibitory concentration (MIC) of GEC was determined based on the relationship between a modified agar diffusion method and a broth microdilution method. GEC inhibited the growth of the Actinomyces, periodontopathic bacteria and Candida strains tested, but did not inhibit the growth of the oral streptococci that are important in the normal oral flora. Commercially available moisture gels containing antimicrobial components showed antimicrobial activity against all of the tested strains. After a series of washes and after a 24-h incubation, GEC retained the antimicrobial activity of the catechins. Catalase prevented GEC-induced growth inhibition of Actinomyces naeslundii and Streptococcus mutans suggesting that hydrogen peroxide may be involved in the antimicrobial activity of catechins. These results suggest that GEC may be useful for controlling oral microorganism populations and reducing the accumulation of dental plaque, thereby helping to prevent periodontal disease and oral candidiasis. PMID:21532150

  17. In vitro antimicrobial findings for fusidic acid tested against contemporary (2008-2009) gram-positive organisms collected in the United States.

    Science.gov (United States)

    Jones, Ronald N; Mendes, Rodrigo E; Sader, Helio S; Castanheira, Mariana

    2011-06-01

    Fusidic acid has a long history of consistent activity against staphylococcal pathogens including methicillin-resistant Staphylococcus aureus (MRSA). Fusidic acid (CEM-102) was susceptibility tested against a surveillance study collection of 12,707 Gram-positive pathogens (2008-2009) from the United States. Reference broth microdilution method results demonstrated the following MIC(50/90) results: S. aureus (.12/.25 μg/mL), coagulase-negative staphylococci (.12/.25 μg/mL), enterococci (4/4 μg/mL), Streptococcus pyogenes (4/8 μg/mL), and viridans group Streptococcus spp. (>8/>8 μg/mL). At a proposed susceptible breakpoint (≤1 μg/mL), fusidic acid inhibited 99.7% of MRSA strains and 99.3% to 99.9% of multidrug-resistant phenotypes of S. aureus. Furthermore, S. aureus strains nonsusceptible to fusidic acid (.35%) generally had detectable resistance mechanisms (fusA, B, C, and E). Reviews of in vitro susceptibility test development confirm the accuracy and intermethod reproducibility of various fusidic acid methods. Fusidic acid is a promising oral therapy for staphylococcal skin and skin structure infections in the United States, where the contemporary S. aureus population remains without significant resistance. PMID:21546624

  18. The antibacterial activity of selected plants towards resistant bacteria isolated from clinical specimens

    Institute of Scientific and Technical Information of China (English)

    Pratiwi Wikaningtyas; Elin Yulinah Sukandar

    2016-01-01

    Objective: To evaluate the antibacterial activity of eight plants against methicillin-resistant Staphylococcus aureus (MRSA), extended spectrum beta-lactamase and carbapenemase-resistant Enterobacteriaceae, which are the most prevalent causes of in-fections in inpatients. Methods: The antibacterial activity was calculated based on the minimum inhibitory concentration using Mueller–Hinton broth in a microdilution method. Results: The best antibacterial activity, calculated as minimum inhibitory concentration values, against MRSA was shown by the Kaempferia pandurata (Roxb) (K. pandurata) extract (256 mg/mL) and the Senna alata (S. alata) extract (512 mg/mL). Phytochemical screening of dried S. alata leaf and its extract showed the presence of flavonoids, alka-loids, saponins, quinones, tannins and sterols, while dried K. pandurata and its extract only showed the presence of flavonoids and sterols/triterpenoids. Conclusions: K. pandurata and S. alata have the potential to be developed as antibac-terial agents, especially against MRSA strain, but further in vivo research and discovery of the mode of its action are still needed to shed light on the effects.

  19. Antifungal activity of selected Malaysian honeys: a comparison with Manuka honey

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    Siti Aisyah Sayadi

    2015-07-01

    Full Text Available Objective: To evaluate four selected Malaysian honey samples from different floral sources (Gelam, Tualang, Nenas and Acacia for their ability to inhibit the growth of fungi and yeast strains (Candida albicans, Aspergillus niger, Epidermophyton floccosum, Microsporum gypseum, Trichophyton rubrum and Trichophyton mentagrophytes. Methods: The broth microdilution method was used to assess the antifungal activity of honey against yeasts at different concentrations ranging from 0.01% to 70% (v/v. Minimum inhibitory concentration (MIC of the honeys were determined by visual inspection and spectrophotometric assay. Minimum fungicidal concentration test was performed by further sub-culturing from the plates which showed no visible growth in the MIC assay onto Sabroud dextrose agar. Results: All tested Malaysian honeys except Gelam showed antifungal activity against all species analysed, with the MIC ranging from 25% (v/v to 50% (v/v while MIC of Manuka honey ranged between 21% to 53% (v/v. Candida albicans was more susceptible to honey than other species tested. Conclusions: Locally produced honeys exhibited antifungal activity which is less than or equal to that of Manuka honey. Our data showed evidence in support of the therapeutic uses of Malaysian honeys.

  20. Antifungal activity of traditional medicinal plants from Tamil Nadu, India

    Institute of Scientific and Technical Information of China (English)

    Duraipandiyan V; Ignacimuthu S

    2011-01-01

    Objective:To assess the antifungal activity of hexane, ethyl acetate and methanol extracts of 45 medicinal plants and to determine the minimum inhibitory concentration for each extract against human pathogenic fungi. Methods:A total of 45 medicinal plants were collected from different places of Tamil Nadu and identified. Hexane, ethyl acetate and methanol extracts of 45 medicinal plants were assessed for antifungal susceptibility using broth microdilution method. Two known antifungal agents were used as positive controls. Results: Most of the extracts inhibited more than four fungal strains. From the evaluation we found that ethyl acetate extracts inhibited large number of fungal growth. Hexane extracts also nearly showed the same level of inhibition against fungal growth. Methanol extracts showed the minimum antifungal activity. Among the 45 plants tested, broad spectrum antifungal activity was detected in Albizzia procera (A. procera), Atalantia monophylla, Asclepias curassavica, Azima tetracantha, Cassia fistula (C. fistula), Cinnomomum verum, Costus speciosus (C. speciosus), Nymphaea stellata, Osbeckia chinensis, Piper argyrophyllum, Punica granatum, Tinospora cordifolia and Toddalia asiatica (T. asiatica). Promising antifungal activity was seen in A. procera, C. speciosus, C. fistula and T. asiatica. Conclusions:It can be concluded that the plant species assayed possess antifungal properties. Further phytochemical research is needed to identify the active principles responsible for the antifungal effects of some of these medicinal plants.

  1. Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae causing noninvasive diseases in a Children's Hospital, Shanghai

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    Fen Pan

    2015-04-01

    Full Text Available Background:Streptococcus pneumoniae, which cause noninvasive pneumococcal diseases, severely impair children's health. This study analyzed serotype distribution and antimicrobial resistance of S. pneumoniae from January 2012 to December 2012 in a Children's Hospital, Shanghai.Methods:A total of 328 pneumococcal isolates were serotyped by multiplex sequential PCR and/or capsule-quellung reaction. The minimum inhibitory concentrations for 11 antimi- crobial agents were determined by broth microdilution method.Results:Among 328 strains, 19F (36.3%, 19A (13.4%, 6A (11.9%, 23F (11.0%, 14 (5.8%, 6B (5.2%, and 15B/C (4.3% were the most common serotypes. The coverage rates of 7-, 10-, and 13-valent conjugate vaccines (PCV7, PCV10, and PCV13 were 58.2%, 58.2%, and 84.1%, respectively. Out of the isolates, 26 (7.9% strains were penicillin resistant. Most of the strains displayed high resistance rate to macrolides (98.5% to erythromycin, 97.9% to azithromycin, and 97.0% to clindamycin.Conclusions:The potential coverage of PCV13 is higher than PCV7 and PCV10 because of the emergence of 19A and there should be long-term and systematic surveillance for non-vaccine serotypes.

  2. β-Aescin at subinhibitory concentration (sub-MIC) enhances susceptibility of Candida glabrata clinical isolates to nystatin.

    Science.gov (United States)

    Franiczek, Roman; Gleńsk, Michał; Krzyżanowska, Barbara; Włodarczyk, Maciej

    2015-11-01

    Aescin (escin) derived from the seeds of horse chestnut (Aesculus hippocastanum L.) is a natural mixture of triterpene saponins exhibiting a wide variety of pharmacological properties, including antiinflammatory, analgesic, and antipyretic activities. However, data concerning antifungal activities of these compounds are limited. This study aims to evaluate the in vitro antifungal susceptibility of Candida glabrata clinical isolates to α-aescin sodium, β-aescin crystalline and β-aescin sodium using the disk diffusion (DD) and broth microdilution (BMD) methods. Moreover, the influence of subinhibitory concentration (0.5×MIC) of β-aescins on the nystatin MIC was also studied. In general, the results obtained by the DD assay correlated well with those obtained by the BMD method. Both β-aescins effectively inhibited the growth of all 24 strains tested. The minimum inhibitory concentration (MIC) values ranging from 8 to 32 μg/ml for β-aescin crystalline, whereas those of β-aescin sodium were slightly lower and ranged from 4 to 16 μg/ml. In contrast, α-aescin sodium was found to be completely ineffective against the strains studied. MIC values of nystatin were reduced 2-16-fold and 2-4-fold in the presence of subinhibitory concentration of β-aescin crystalline and β-aescin sodium, respectively. Results of the present study may suggest the additive interaction between β-aescin and nystatin. PMID:26092104

  3. Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing—a continuous attempt at species and serovar differentiation

    Science.gov (United States)

    Moreno, Luisa Z; Miraglia, Fabiana; Lilenbaum, Walter; Neto, José SF; Freitas, Julio C; Morais, Zenaide M; Hartskeerl, Rudy A; da Costa, Barbara LP; Vasconcellos, Silvio A; Moreno, Andrea M

    2016-01-01

    Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing. PMID:26956446

  4. Antibacterial and antioxidant activities in extracts of fully grown cladodes of 8 cultivars of cactus pear.

    Science.gov (United States)

    Sánchez, E; Dávila-Aviña, J; Castillo, S L; Heredia, N; Vázquez-Alvarado, R; García, S

    2014-04-01

    The antimicrobial and antioxidant activities of some cultivars of the nopal cactus have not been determined. In this study, 8 cultivars of nopal cacti from Mexico were assayed for phenolic content, antioxidant activities, and antimicrobial activities against Campylobacter Jejuni, Vibrio cholera, and Clostridium Perfringens. Plant material was washed, dried, and macerated in methanol. Minimum bactericidal concentrations (MBCs) were determined using the broth microdilution method. Antioxidant activities were quantitatively determined using spectrophotometric methods. The MCBs of the nopal cacti ranged from 1.1 to 12.5 mg/mL for c. jejuni, 4.4 to 30 mg/mL for V. cholera, and 0.8 to 16 mg/mL for C. perfringens in the cultivars Cardon Blanco, Real de Catorce, and Jalpa, respectively. High quantities of total phenols and total flavonoids were found in the Jalpa cacti (3.80 mg of gallic acid equivalent GAE/g dry weight [DW] and 36.64 mg of quercetin equivalents [QE]/g DW, respectively). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities (RSA) were correlated to bioactive compound contents. The Villanueva cacti had the highest %RSA at 42.31%, and the lowest activity was recorded in Copena V1 at 19.98%. In conclusion, we found that some of the 8 cactus pear cultivars studied may be used for their antioxidant compounds or antimicrobials to control or prevent the contamination of foods. PMID:24621296

  5. Charcterization of Type Ⅱ Topoisomerase Gene Mutations in Clinical Isolates of Mycoplasma Hominis Resistant to Fluoroquinolones

    Institute of Scientific and Technical Information of China (English)

    吴移谋; 张文波; 姚艳冰

    2002-01-01

    Objective: To analyze type Ⅱ topoisomerase genes inclinical isolates of fluoroquinolone-resistant Mycoplasmahominis. Methods: Eight isolates of M.hominis cross resistant to 6fluoroquinolones were selected from 103 clinical strains ofM.hominis using a broth microdilution method. Type IItopoisomerase genes were amplified by PCR and directlysequenced. Nucleotide sequences were compared to sequencesfrom a susceptible strain (M.hominis PG2I). Results: MICs of resistant Mh isolates were 4- to 512-fold higher than MICs from the susceptible reference strain.Sequence comparison revealed a C to T change at 113nt ingyrA QRDR led to the substitution of Ser83 by Leucine and noamino acid change in gyrB. A change of G to T at 134nt inparC QRDR led to the substitution of Ser80 by Isoleucine anda G to A change at 70nt in parE QRDR led to the substitutionof Aspartic acid by Asparagine. Conclusion: These results suggest that a C to T change atll3nt in gyrA, a G to T change at 134nt in parC and a G to Achange at 70nt in patrE are associated with fluoroquinoloneresistance of M.hominis.

  6. Study of Antibacterial Effect of Novel Thiazole, Imidazole and Tetrahydropyridine Derivatives against Escherichia coli

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    Behzad Ghasemi

    2016-01-01

    Full Text Available > Introduction: Escherichia coli is one of the important pathogens in human with globalimportance. Because of the necessity for identification and the use of novel antibacterialcompounds against E. coli, in this present study we focused on the antibacterial effects ofsynthesized thiazole, imidazole and tetrahydropyridine derivatives on E. coli.Methods: For evaluation of antibacterial effect, the disk diffusion method was applied to measurethe growth inhibition zone diameter and broth micro-dilution was performed to determine theminimum inhibitory concentration (MIC.Results: Assessing the antibacterial effect showed that only 6d derivative of thiazole hadinhibitory effect on E. coli and the other thiazole, imidazole and tetrahydropyridine derivativeslacked any inhibitory result on this organism. The inhibitory effect of 6d derivative of thiazolewas MIC=125 and growth inhibition zone diameter of 16±0.1.Discussion: The antibacterial effect of thiazole, imidazole and tetrahydropyridine derivativesdiffers from each other and chemical linkages such as oxygen to thiazole ring in 6d derivative,could have reinforced this effect. The next step is determination of the toxicity and therapeuticeffects in the laboratory animals.

  7. Evaluation of anti-bacterial and anti-oxidant potential of andrographolide and echiodinin isolated from callus culture of Andrographis paniculata Nees

    Institute of Scientific and Technical Information of China (English)

    Mohammed Arifullah; Nima Dandu Namsa; Manabendra Mandal; Kishore Kumar Chiruvella; Paritala Vikrama; Ghanta Rama Gopal

    2013-01-01

    Objective: To evaluate the anti-bacterial and anti-oxidant activity of andrographolide (AND) and echiodinin (ECH) of Andrographis paniculata. Methods:In this study, an attempt has been made to demonstrate the anti-microbial and anti-oxidant activity of isolated AND and ECH by broth micro-dilution method and 2,2-diphenyl-2-picryl-hydrazyl (DPPH) assay, respectively. Structure elucidation was determined by electro-spray ionization-MSD, NMR (1H and 13C) and IR spectra. Results: AND was effective against most of the strains tested including Mycobacteriumsmegmatis, showing broad spectrum of growth inhibition activity with Minimum inhibitory concentration values against Staphylococcus aureus (100 µg/mL), Streptococcus thermophilus (350 µg/mL) Bacillus subtilis (100 µg/mL), Escherichia coli (50 µg/mL), Mycobacterium smegmatis (200 µg/mL), Klebsiella pneumonia (100 µg/mL), and Pseudomonas aeruginosa (200 µg/mL). ECH showed specific anti-bacterial activity against Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa at a concentration higher than 225 µg/mL. Both AND and ECH were not effective against the two yeast strains, Candida albicans and Saccharomyces cerevisiae tested in this study. Conclusion:This preliminary study showed promising anti-bacterial activity and moderate free radical scavenging activity of AND and ECH, and it may provide the scientific rationale for its popular folklore medicines.

  8. Determination of Nitrate Reductase Assay Depending on the Microbial Growth

    International Nuclear Information System (INIS)

    A rapid micro-dilution assay for determination of the antimicrobial susceptibility of different bacterial isolates was developed. This assay is based on the ability of the most of viable organisms to reduce nitrates. The MIC or MBC could be determined by nitrate reductase (NR) only after 30 to 90 min of incubation depending on the behaviour of microbial growth. Bacterial viability is detected by a positive nitrite reduction rather than visible turbidity. The nitrate reduction assay was compared with standard micro-assay using 250 isolates of different taxa against 10 antibiotics belonging to different classes. An excellent agreement of 82.5 % was found between the two methods and only 17.5 % of 1794 trials showed difference in the determined MIC by tow-dilution interval above or below the MIC determined by the turbidimetric method under the same test conditions. However, the nitrate reduction assay was more rapid and sensitive in detecting viable bacteria and so, established an accurate estimate of the minimal inhibitory concentration (MIC) or the minimal bacterial concentration (MBC). The nitrate reduction assay offers the additional advantage that it could be used to determine the MBC without having to subculture the broth. 232 cases of resistance were detected by NR and 4 different media were tested for susceptibility test. The bacterial isolates were exposed to ultra violet (UV) light for different period

  9. Comparative study on the antifungal activity of hydroalcoholic extract of Iranian Propolis and Royal jelly against Rhizopus oryzae

    Directory of Open Access Journals (Sweden)

    Moghim Hassan

    2015-07-01

    Full Text Available Introduction: Mucormycosis is an opportunistic fungal infection. Rhizopus oryzae is major cause of mucormycosis in humans. This disease is the most common form of the acute fungal infection with rapid progress. Iranian Propolis extract and Royal jelly are honey bee products which have been used by human over the past centuries in traditional medicine. In this study the effects of Iranian Propolis and Royal jelly were investigated against Rhizopus oryzae and Candida albicans. Methods: The used method in this study was microdilution. To perform it, the prepared dilutions of Royal jelly and alcoholic extract of Iranian Propolis were added to tubes containing Sabouraud dextrose broth culture media except to control group and then Rhizopus oryzae suspension was added to all microtubes. In the next stage, microtubes were maintained in incubator at 25oC for 48 hours and then 10 ml of the content of each microtube was transferred to Sabouraud dextrose agar media. The minimum inhibitory concentration (MIC and minimum fungicidal concentration (MFC of the agents were calculated. Results: In this study, the MIC and MFC of Iranian Propolis alcoholic extract on Rhizopus oryzae were respectively 0.1 and 0.25 mg/ml and the MFC of Royal jelly on Rhizopus oryzae were respectively 100 ± 34 and 133 ± 46 mg/ml. Conclusion: The results indicate that Propolis might be used as an ideal combination for the treatment of fungal infections like Rhizopus oryzae. However, clinical studies are needed to confirm the effects of these drugs.

  10. Antimicrobial activity of some essential oils against oral multidrug-resistant Enterococcus faecalis in both planktonic and biofilm state

    Science.gov (United States)

    Benbelaïd, Fethi; Khadir, Abdelmounaïm; Abdoune, Mohamed Amine; Bendahou, Mourad; Muselli, Alain; Costa, Jean

    2014-01-01

    Objective To evaluate some essential oils in treatment of intractable oral infections, principally caused by biofilm of multidrug-resistant Enterococcus faecalis (E. faecalis), such as persistent endodontic infections in which their treatment exhibits a real challenge for dentists. Methods Ten chemically analyzed essential oils by gas chromatography-mass spectrometry were evaluated for antimicrobial activity against sensitive and resistant clinical strains of E. faecalis in both planktonic and biofilm state using two methods, disk diffusion and broth micro-dilution. Results Studied essential oils showed a good antimicrobial activity and high ability in E. faecalis biofilm eradication, whether for sensitive or multidrug-resistant strains, especially those of Origanum glandulosum and Thymbra capitata with interesting minimum inhibitory concentration, biofilm inhibitory concentration, and biofilm eradication concentration values which doesn't exceed 0.063%, 0.75%, and 1.5%, respectively. Conclusions Findings of this study indicate that essential oils extracted from aromatic plants can be used in treatment of intractable oral infections, especially caused by biofilm of multidrug-resistant E. faecalis. PMID:25182948

  11. Synergic antibacterial activity of some essential oils from Lamiaceae

    Directory of Open Access Journals (Sweden)

    Sh. Fahimi

    2015-05-01

    Full Text Available Background and objectives: Despite the vast production of new antibiotics in the last three decades, resistance to these drugs by microorganisms has increased and essential oils (EOs have been recognized to possess antimicrobial properties. Methods:  In the present study, EOs obtained from aerial parts of Thymus vulgaris L., Lavandula angustifolia Mill., Rosmarinus officinalis L. and Mentha piperita L., were evaluated for their single and binary combined antibacterial activities against four Gram-positive and Gram-negative pathogenic bacteria: Staphylococcus aureus, Bacillus cereus, Escherichia coli and Pseudomonas aeruginosa. Results: The results exhibited that some of the tested essential oils revealed antibacterial activities against the examined pathogens using broth microdilution method. Maximum activity of the testedessential oils was obtained from the combination of T. vulgaris and M. piperita essential oils against Staphylococcus aureus (MIC= 0.625 mg/mL. Conclusion: Combinations of the essential oils in this study showed synergic action against some pathogenic microorganisms which could be considered in medical and food industries as preservatives.

  12. New tridentate azo-azomethines and their copper(II) complexes: Synthesis, solvent effect on tautomerism, electrochemical and biological studies

    Science.gov (United States)

    Sarigul, Munire; Deveci, Pervin; Kose, Muhammet; Arslan, Ugur; Türk Dagi, Hatice; Kurtoglu, Mukerrem

    2015-09-01

    In this study, three azo-azomethines and their copper(II) complexes were prepared and characterized by analytical and spectroscopic methods. The complexes prepared were found to be mononuclear and the chelation of the ligands to the copper(II) ions occurs through two phenolic oxygens and a nitrogen atom of the azomethine group of the ligand. The tautomeric behaviors of the azo-azomethines in solution were studied by UV-Vis. spectra in three organic solvents with different polarity (CHCl3, DMSO and DMF) at room temperature. The redox behaviors of the azo-azomethines and their Cu(II) complexes were investigated by cyclic voltammetry (CV) in DMSO solution containing 0.1 M tetrabutylammonium tetrafluoroborate (TBATFB) as supporting electrolyte. Additionally, the antibacterial activity was also evaluated by the broth microdilution methods against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. The compounds were found to be less effective against all bacteria tested than two reference antibiotics (ampicillin and gentamicin).

  13. Antimicrobial activity of Amazonian oils against Paenibacillus species.

    Science.gov (United States)

    Santos, Roberto Christ Vianna; dos Santos Alves, Camilla Filippi; Schneider, Taiane; Lopes, Leonardo Quintana Soares; Aurich, Carlos; Giongo, Janice Luehring; Brandelli, Adriano; de Almeida Vaucher, Rodrigo

    2012-03-01

    The Gram-positive, spore-forming bacterium Paenibacillus larvae is the primary bacterial pathogen of honeybee brood and the causative agent of American foulbrood disease (AFB). One of the feasible alternative treatments being used for their control of this disease is essential oils. In this study in vitro antimicrobial activity of Andiroba and Copaíba essential oils against Paenibacillus species, including P. larvae was evaluated. Minimal inhibitory concentration (MIC) in Mueller-Hinton broth by the microdilution method was assessed. Andiroba registered MIC values of 1.56-25%, while the MICs values obtained for Copaíba oil were of 1.56-12.5%. In order to determine the time-response effect of essential oils on P. larvae, this microorganism was exposed to the oils for up to 48 h. After 24 h treatment with Andiroba oil and after 48 h treatment with Copaíba oil no viable cells of P. larvae ATCC 9545 were observed. The possible toxic effect of essential oils were assessed by the spraying application method of the same concentrations of MICs. Bee mortality was evident only in treatment with Andiroba oil and the Copaíba oil shows no toxic effects after 10 days of observation. Taking together ours results showed for the first time that these oils presented a high activity against Paenibacillus species showing that Copaíba oil may be a candidate for the treatment or prevention of AFB. PMID:22200645

  14. Combination Therapy of Sophoraflavanone B against MRSA: In Vitro Synergy Testing.

    Science.gov (United States)

    Mun, Su-Hyun; Kang, Ok-Hwa; Joung, Dae-Ki; Kim, Sung-Bae; Seo, Yun-Soo; Choi, Jang-Gi; Lee, Young-Seob; Cha, Seon-Woo; Ahn, Young-Sup; Han, Sin-Hee; Kwon, Dong-Yeul

    2013-01-01

    Sophoraflavanone B (SPF-B), a known prenylated flavonoid, was isolated from the roots of Desmodium caudatum. The aim of this study was to determine the antimicrobial synergism of SPF-B combined with antibiotics against methicillin-resistant Staphylococcus aureus (MRSA). MRSA, a multidrug-resistant pathogen, causes both hospital- and community-acquired infections worldwide. The antimicrobial activity of SPF-B was assessed by the broth microdilution method, checkerboard dilution test, and time-kill curve assay. The MIC of SPF-B for 7 strains of S. aureus ranges from 15.6 to 31.25  μ g/mL determined. In the checkerboard method, the combinations of SPF-B with antibiotics had a synergistic effect; SPF-B markedly reduced the MICs of the β -lactam antibiotics: ampicillin (AMP) and oxacillin (OXI); aminoglycosides gentamicin (GET); quinolones ciprofloxacin (CIP) and norfloxacin (NOR) against MRSA. The time-kill curves assay showed that a combined SPF-B and selected antibiotics treatment reduced the bacterial counts below the lowest detectable limit after 24 h. These data suggest that the antibacterial activity of SPF-B against MRSA can be effectively increased through its combination with three groups of antibiotics ( β -lactams, aminoglycosides, and quinolones). Our research can be a valuable and significant source for the development of a new antibacterial drug with low MRSA resistance. PMID:24319486

  15. Anti-Helicobacter pylori activity and immunostimulatory effect of extracts from Byrsonima crassa Nied. (Malpighiaceae

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    Vilegas Wagner

    2009-01-01

    Full Text Available Abstract Background Several in vitro studies have looked at the effect of medicinal plant extracts against Helicobacter pylori (H. pylori. Regardless of the popular use of Byrsonima crassa (B. crassa as antiemetic, diuretic, febrifuge, to treat diarrhea, gastritis and ulcers, there is no data on its effects against H. pylori. In this study, we evaluated the anti-H. pylori of B. crassa leaves extracts and its effects on reactive oxygen/nitrogen intermediates induction by murine peritoneal macrophages. Methods The minimal inhibitory concentration (MIC was determined by broth microdilution method and the production of hydrogen peroxide (H2O2 and nitric oxide (NO by the horseradish peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. Results The methanolic (MeOH and chloroformic (CHCl3 extracts inhibit, in vitro, the growth of H. pylori with MIC value of 1024 μg/ml. The MeOH extract induced the production H2O2 and NO, but CHCl3 extract only NO. Conclusion Based in our results, B. crassa can be considered a source of compounds with anti-H. pylori activity, but its use should be done with caution in treatment of the gastritis and peptic ulcers, since the reactive oxygen/nitrogen intermediates are involved in the pathogenesis of gastric mucosal injury induced by ulcerogenic agents and H. pylori infections.

  16. Antibacterial activity of Thymus vulgaris essential oil alone and in combination with other essential oils

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    MAHENDRA RAI

    2012-07-01

    Full Text Available Kon K, Rai M. 2012. Antibacterial activity of Thymus vulgaris essential oil alone and in combination with other essential oils. Nusantara Bioscience 4: 50-56. Essential oils (EOs from plants represent an alternative approach in combating antibiotic-resistant bacteria. One of the EOs with proven antibacterial properties is Thymus vulgaris EO. The purpose of the present work was to investigate in vitro antibacterial activity of T. vulgaris EO alone and in combination with other EOs. The activity of T. vulgaris EO was screened in combination with 34 EOs against Staphylococcus aureus and Escherichia coli by disk diffusion method; then the most effective combinations were evaluated by broth microdilution method. Against S. aureus the synergistic effect was found in combination of T. vulgaris and Cinnamomum zeylonicum EOs with fractional inhibitory concentration (FIC index of 0.26; Juniperus communis and Picea abies EOs showed additive effect (FIC indexes were 0.74 and 0.78, respectively. Combination of T. vulgaris EO with Aniba rosaeodora and Melissa officinalis EOs demonstrated synergistic effect against E. coli (FIC indexes were 0.23 and 0.34, respectively; combination of T. vulgaris and Mentha piperita EOs was additive (FIC index 0.55. Therefore, combining T. vulgaris EO with other EOs has potential in further enhancing its antibacterial properties.

  17. Profiling of Leptospira interrogans, L. santarosai, L. meyeri and L. borgpetersenii by SE-AFLP, PFGE and susceptibility testing-a continuous attempt at species and serovar differentiation.

    Science.gov (United States)

    Moreno, Luisa Z; Miraglia, Fabiana; Lilenbaum, Walter; Neto, José Sf; Freitas, Julio C; Morais, Zenaide M; Hartskeerl, Rudy A; da Costa, Barbara Lp; Vasconcellos, Silvio A; Moreno, Andrea M

    2016-01-01

    Leptospirosis is a widespread systemic zoonosis, considered as reemerging in certain developing countries. Although the cross agglutinin absorption test is still considered the standard method for Leptospira identification, it presents several disadvantages. The aim of this study was to characterize Leptospira spp. isolated from various hosts by genotyping and broth microdilution susceptibility testing in an attempt to differentiate Leptospira species, serogroups and serovars. Forty-seven isolates were studied. They were previously serotyped, and species confirmation was performed by 16S rRNA sequencing. Single-enzyme amplified fragment length polymorphism (SE-AFLP) and pulsed-field gel electrophoresis (PFGE) analysis enabled the distinction of L. interrogans from L. santarosai, L. meyeri and L. borgpetersenii in two main clusters. Among L. interrogans, it was possible to differentiate into two new clusters the serogroup Icterohaemorrhagiae from the serogroups Canicola and Pomona. L. santarosai isolates presented higher genetic variation than the other species in both techniques. Interestingly, the minimum inhibitory concentration (MIC) cluster analysis also provided Leptospira serogroup differentiation. Further studies are necessary regarding serovar Bananal isolates, as they presented the highest MIC values for most of the antimicrobials tested. All studied techniques successfully distinguished Leptospira species and serogroups. Despite being library-dependent methods, these approaches are less labor intensive and more economically viable, particularly SE-AFLP, and can be implemented in most reference laboratories worldwide to enable faster Leptospira typing. PMID:26956446

  18. A rapid fluorescence based method for the quantitative analysis of cell culture media photo-degradation.

    Science.gov (United States)

    Calvet, Amandine; Li, Boyan; Ryder, Alan G

    2014-01-01

    Cell culture media are very complex chemical mixtures that are one of the most important aspects in biopharmaceutical manufacturing. The complex composition of many media leads to materials that are inherently unstable and of particular concern, is media photo-damage which can adversely affect cell culture performance. This can be significant particularly with small scale transparent bioreactors and media containers are used for process development or research. Chromatographic and/or mass spectrometry based analyses are often time-consuming and expensive for routine high-throughput media analysis particularly during scale up or development processes. Fluorescence excitation-emission matrix (EEM) spectroscopy combined with multi-way chemometrics is a robust methodology applicable for the analysis of raw materials, media, and bioprocess broths. Here we demonstrate how EEM spectroscopy was used for the rapid, quantitative analysis of media degradation caused by ambient visible light exposure. The primary degradation pathways involve riboflavin (leading to the formation of lumichrome, LmC) which also causes photo-sensitised degradation of tryptophan, which was validated using high pressure liquid chromatography (HPLC) measurements. The use of PARallel FACtor analysis (PARAFAC), multivariate curve resolution (MCR), and N-way partial least squares (NPLS) enabled the rapid and easy monitoring of the compositional changes in tryptophan (Trp), tyrosine (Tyr), and riboflavin (Rf) concentration caused by ambient light exposure. Excellent agreement between HPLC and EEM methods was found for the change in Trp, Rf, and LmC concentrations. PMID:24356227

  19. Antibacterial activity of three medicinal Thai plants against Campylobacter jejuni and other foodborne pathogens.

    Science.gov (United States)

    Dholvitayakhun, Achara; Cushnie, T P Tim; Trachoo, Nathanon

    2012-01-01

    Leaves of Adenanthera pavonina, Moringa oleifera and Annona squamosa are used in traditional Thai medicine to treat dysentery and other diseases. This study investigated the antibacterial activity of these plants against six species of foodborne pathogen. Methods and solvents employed to extract active constituents were optimised using the disc diffusion assay. Phytochemical analysis of the optimised extracts was performed by thin layer chromatography (TLC). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by broth microdilution. A. pavonina contained flavonoids, terpines and tannins, and was the most active extract against Campylobacter jejuni, inhibiting growth at 62.5-125 µg mL(-1). The A. squamosa extract contained flavonoids, terpines, tannins and alkaloids, and had the broadest spectrum of antibacterial activity, inhibiting Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus and C. jejuni between 62.5 and 500 µg mL(-1). MBCs were 2- to 4-fold higher than MICs against C. jejuni and B. cereus, suggesting the extracts are bactericidal against these species. Negligible activity was detected from M. oleifera. The data presented here show that A. pavonina and A. squamosa could potentially be used in modern applications aimed at the treatment or prevention of foodborne diseases. PMID:21878033

  20. Simvastatin inhibits planktonic cells and biofilms ofCandida and Cryptococcusspecies

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    Raimunda Sâmia Nogueira Brilhante

    2015-10-01

    Full Text Available ABSTRACTThe antifungal activity of some statins against different fungal species has been reported. Thus, at the first moment, the in vitro antifungal activity of simvastatin, atorvastatin and pravastatin was tested againstCandida spp. and Cryptococcus spp. Then, in a second approach, considering that the best results were obtained for simvastatin, this drug was evaluated in combination with antifungal drugs against planktonic growth and tested against biofilms ofCandida spp. and Cryptococcus spp. Drug susceptibility testing was performed using the microdilution broth method, as described by the Clinical and Laboratory Standards Institute. The interaction between simvastatin and antifungals against planktonic cells was analyzed by calculating the fractional inhibitory concentration index. Regarding biofilm susceptibility, simvastatin was tested against growing biofilm and mature biofilm of one strain of each tested yeast species. Simvastatin showed inhibitory effect against Candida spp. andCryptococcus spp. with minimum inhibitory concentration values ranging from 15.6 to 1000 mg L-1 and from 62.5 to 1000 mg L-1, respectively. The combination of simvastatin with itraconazole and fluconazole showed synergism against Candidaspp. and Cryptococcus spp., while the combination of simvastatin with amphotericin B was synergistic only againstCryptococcus spp. Concerning the biofilm assays, simvastatin was able to inhibit both growing biofilm and mature biofilm ofCandida spp. and Cryptococcus spp. The present study showed that simvastatin inhibits planktonic cells and biofilms ofCandida and Cryptococcus species.

  1. Green synthesis of silver nanoparticles as antibacterial agent using Rhodomyrtus tomentosa acetone extract

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    Voravuthikunchai, Supayang P.; Chorachoo, Julalak; Jaiswal, Lily; Shankar, Shiv

    2013-12-01

    The capability of Rhodomyrtus tomentosa acetone extract (RAE) for the production of silver nanoparticles (AgNPs) has been explored for the first time. Silver nanoparticles with a surface plasmon resonance band centered at 420-430 nm were synthesized by reacting RAE with AgNO3. Reaction time, temperature, concentration of AgNO3 and RAE could accelerate the reduction rate of Ag+ and affect AgNPs size. The nanoparticles were found to be 10-30 nm in size and spherical in shape. XRD data demonstrated crystalline nature of AgNPs dominated by (200) facets. FTIR results showed decrease in intensity of peaks at 3394, 1716 and 1618 cm-1 indicating the involvement of O-H, carbonyl group and C=C stretching with the formation of AgNPs with RAE, respectively. The C-O-C and C-N stretching suggested the presence of many phytochemicals on the surface of the nanoparticles. High negative zeta potential values confirmed the stability of AgNPs in water. In vitro antibacterial activity of AgNPs was tested against Staphylococcus aureus using broth microdilution method. AgNPs capped with RAE demonstrated profound antibacterial activity against the organisms with minimum inhibitory concentration and minimum bactericidal concentration in the range between 3.1-6.2 and 6.2-50 μgmL-1, respectively. The synthesized nanoparticles could be applied as an effective antimicrobial agent against staphylococcal infections.

  2. Chemical constituents and antimicrobial properties of the essential oil and ethanol extract from the stem of Aglaia odorata Lour.

    Science.gov (United States)

    Joycharat, Nantiya; Thammavong, Sonesay; Voravuthikunchai, Supayang Piyawan; Plodpai, Patimaporn; Mitsuwan, Watcharapong; Limsuwan, Surasak; Subhadhirasakul, Sanan

    2014-01-01

    The stem-derived essential oil of Aglaia odorata Lour. was obtained by hydrodistillation using a Clevenger-type system. Gas chromatography-mass spectrometry analysis of the oil revealed the identification of 39 compounds, representing 76.4% of the oil; germacrene D (20.3%), α-humulene (17.1%), α-himachalene (12.7%) and β-caryophyllene (10.2%) were the major components. Ar-turmerone (1) and eichlerialactone (2) were isolated from the stem oil and ethanolic stem extract of this plant species, respectively. Antimicrobial activities of the oil and ethanol extract were tested against both Gram-positive and Gram-negative bacterial strains including Bacillus cereus ATCC 11778, Staphylococcus aureus ATCC 25923, Acinetobacter baumannii ATCC 19606 and Escherichia coli ATCC 25922, as well as three rice fungal pathogens Bipolaris oryzae, Pyricularia oryzae and Rhizoctonia solani using broth microdilution method. The oil and 1 exhibited significant antifungal activity against the three rice pathogens tested, whereas 2 exhibited good antibacterial activity against both the Gram-positive pathogens tested. PMID:24934340

  3. Antifungal Properties of Chenopodium ambrosioides Essential Oil Against Candida Species

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    Gerard Vilarem

    2010-09-01

    Full Text Available The essential oil of the aerial part (leaves, flowers and stem of Chenopodium ambrosioides was obtained by hydrodistillation and its chemical composition analyzed by GC and GC/MS, which permitted the identification of 14 components, representing 98.8% of the total oil. Major components were α-terpinene (51.3%, p-cymene (23.4% and p-mentha-1,8-diène (15.3%. The antifungal properties of this essential oil were investigated in vitro by the well diffusion and broth microdilution methods. The in vitro antifungal activity was concentration dependent and minimum inhibitory concentration values varied from 0.25 to 2 mg/mL. The in vivo antifungal activity was evaluated on an induced vaginal candidiasis rat model. The in vivo activity of the oil on mice vaginal candidiasis was not dose-dependent. Indeed, all the three tested doses; 0.1%, 1% and 10% led to the recovery of mice from the induced infection after 12 days of treatment. The effect of the essential oil on C. albicans ATCC 1663 fatty acid profile was studied. This oil has a relatively important dose-dependent effect on the fatty acids profile.

  4. Evaluation of polysaccharides content in fruit bodies and their antimicrobial activity of four Ganoderma lucidum (W Curt.: Fr. P. Karst. strains cultivated on different wood type substrates

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    Krystyna Skalicka-Woźniak

    2012-03-01

    Full Text Available Quantitative determination of polysaccharides in Ganoderma lucidum fruit bodies from different sawdust cultivation substrates and their antibacterial activity was done. Thirty six samples were analyzed. Four strains of Ganoderma lucidum (GL01, GL02, GL03 and GL04 were cultivated on the growth substrates of three different sawdust types: birch (Bo, maple (Kl or alder (Ol amended with wheat bran in three different concentrations: 10, 20 and 30% (w/w. Even though the richest in polysaccharides was GL01 strain, the highest yields of the polysaccharides were determined in GL04Kl3 sample and was 112.82 mg/g of dry weight. The antibacterial activity of polysaccharides was determined in vitro using micro-dilution broth method. The panel of eight reference bacterial strains was used. All the polysaccharide samples tested showed the broad spectrum and the moderate antibacterial activity. Micrococcus luteus ATCC 10240 strain was the most sensitive with MIC (minimal inhibitory concentration = 0.63 − 1.25 mg/mL.

  5. Antibacterial and Synergy of Berberines with Antibacterial Agents against Clinical Multi-Drug Resistant Isolates of Methicillin-Resistant Staphylococcus aureus (MRSA

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    Zhong-Qi Bian

    2012-08-01

    Full Text Available Antibacterial activity of berberine (Ber and 8-acetonyl-dihydroberberine (A-Ber alone and combined uses with antibacterial agents ampicillin (AMP, azithromycin (AZM, cefazolin (CFZ and levofloxacin (LEV was studied on 10 clinical isolates of SCCmec III type methicillin-resistant Staphylococcus aureus (MRSA. Susceptibility to each agent alone was tested using a broth microdilution method and the chequerboard and time-kill tests for the combined evaluations, respectively. The alone MICs/MBCs (mg/mL ranges were 32–128/64–256 (Ber and 32-128/128-512 (A-Ber. Significant synergies were observed for the Ber (A-Ber/AZM and Ber (A-Ber/LEV combinations against 90% of the tested MRSA strains, with fractional inhibitory concentration indices (FICIs values ranged  from 0.188 to 0.500. An additivity result was also observed for the Ber/AZM combination by time-kill curves. These results demonstrated for the first time that Ber and A-Ber enhanced the in vitro inhibitory efficacy of AZM and LEV to a same extent, which had potential for further investigation in combinatory therapeutic applications of patients infected with MRSA.

  6. Chemical Composition and In Vitro Antibacterial Activity of Mentha spicata Essential Oil against Common Food-Borne Pathogenic Bacteria

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    Yasser Shahbazi

    2015-01-01

    Full Text Available The aim of the present study was to investigate chemical composition and antibacterial activity of essential oil from the leaf of Mentha spicata plant against common food-borne pathogenic bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7. Chemical composition of the essential oil was identified by gas chromatography coupled with mass spectrometer detector (GC-MS. The antibacterial activity of the essential oil was evaluated by broth microdilution method and agar disk diffusion assay. According to the result of GC-MS analysis, 18 components were identified, accounting for 99.89% of the whole essential oil. The main components were carvone (78.76%, limonene (11.50%, β-bourbonene (11.23%, cis-dihydrocarveol (1.43%, trans-caryophyllene (1.04%, menthone (1.01%, menthol (1%, and terpinen-4-ol (0.99. The essential oil exhibited moderate level of antibacterial activity against all test microorganisms. In general, Gram-positive bacteria were more susceptible to M. spicata essential oil than Gram-negative bacteria. L. monocytogenes was the most sensitive of the microorganisms to the antibacterial activity of M. spicata essential oil (inhibition zone = 22 mm and MIC and MBC = 2.5 µL/mL. Based on our results, the essential oil of M. spicata plant collected from Kermanshah province, west of Iran, has a potential to be applied as antibacterial agent.

  7. Efficacy of Cuminum cyminum essential oil on FUM1 gene expression of fumonisin-producing Fusarium verticillioides strains

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    Ali Reza Khosravi

    2014-12-01

    Full Text Available Objectives: The purpose of this study was to evaluate the effect of Cuminum cyminum (C. cyminum essential oil on the growth and FUM1 gene expression of fumonisin-producing Fusarium verticillioides (F. verticillioides strains isolated from maize. Materials and Methods: All fungal strains were cultured on potato dextrose agar (PDA slopes at 30°C for 7 days. The antifungal activity was evaluated by broth microdilution assay. One set of primers was F. verticillioides species specific, which selectively amplified the intergenic space region of rDNA. The other set of primers was specific to FUM1 gene region of fumonisin-producing F. verticillioides. FUM1 transcript levels were quantified using a reverse transcription-polymerase chain reaction (RT-PCR protocol. Results: Minimum inhibitory concentration (MIC values of C. cyminum oil against F. verticillioides strains varied from 0.195 to 0.781 µl.ml-1 (mean MIC value: 0.461 µl.ml-1 indicating 54.5% of the fungal strains inhibited at 0.390 µl.ml-1. PCR analysis of FUM1 gene expression revealed that DNA fragment of 183 bp was amplified in all the isolates of F. verticillioides before treatment with C. cyminum essential oil. Based on RT-PCR analyses, reduction in the expression of fumonisin biosynthetic genes was significant only for FUM1 gene (p

  8. Multiplex PCR for the Rapid Simultaneous Speciation and Detection of Methicillin-Resistance and Genes Encoding Toxin Production in Staphylococcus aureus.

    Science.gov (United States)

    Jones, M E; Köhrer, K; Schmitz, F J

    2001-01-01

    The use of polymerase chain reaction (PCR) to rapidly amplify target DNA molecules has evolved a place within diagnostic microbiology allowing the sensitive and specific identification of microorganisms concurrent with the detection of specific genes involved in resistance or virulence. This is particularly helpful when dealing with problem pathogens such as Staphylococcus aureus, where the rapid diagnosis and detection of methicillin-resistance can assist in the fast implementation of appropriate treatment and infection control measures (1-4). In addition, the rapid assessment of toxin production can ascertain whether toxin-related symptoms need to be considered (5). The protocol described here enables the simultaneous differentiation between S. aureus (and coagulase negative staphylococci) from other eubacterial organisms. Concomitant with species identification this protocol allows for the detection of specific staphylococcal resistance genes, such as mecA, encoding methicillin resistance, or staphylococcal virulence genes, such as seb, sec1 and tst, encoding enterotoxin B, enterotoxin C, and toxic shock syndrome toxin-1, respectively. In S. aureus the detection of methicillin-resistance using agar diffusion or broth micro-dilution techniques, or toxin production using immunodiffusion, agglutination, or ELISA is often difficult. This is because the mecA gene and the genes involved in toxin production may be poorly expressed and influenced significantly by culture conditions (5-10). Other methods, in particular various hybridization techniques have been used as diagnostic tools. However. PMID:21374410

  9. Effects of osmotic pressure, acid, or cold stresses on antibiotic susceptibility of Listeria monocytogenes.

    Science.gov (United States)

    Al-Nabulsi, Anas A; Osaili, Tareq M; Shaker, Reyad R; Olaimat, Amin N; Jaradat, Ziad W; Zain Elabedeen, Noor A; Holley, Richard A

    2015-04-01

    Prevalence of antibiotic resistance of Listeria monocytogenes isolated from a variety of foods has increased in many countries. L. monocytogenes has many physiological adaptations that enable survival under a wide range of environmental stresses. The objective of this study was to evaluate effects of osmotic (2, 4, 6, 12% NaC), pH (6, 5.5, 5.0) and cold (4 °C) stresses on susceptibility of three isolates of L. monocytogenes towards different antibiotics. The minimal inhibitory concentrations (MICs) of tested antibiotics against unstressed (control), stressed or post-stressed L. monocytogenes isolates (an ATCC strain and a meat and dairy isolate) were determined using the broth microdilution method. Unstressed cells of L. monocytogenes were sensitive to all tested antibiotics. In general, when L. monocytogenes cells were exposed to salt, cold and pH stresses, their antibiotic resistance increased as salt concentration increased to 6 or 12%, as pH was reduced to pH 5 or as temperature was decreased to 10 °C. Results showed that both meat and dairy isolates were more resistant than the ATCC reference strain. Use of sub-lethal stresses in food preservation systems may stimulate antibiotic resistance responses in L. monocytogenes strains. PMID:25475279

  10. Antimicrobial Activities of Hexane Extract and Decussatin from Stembark Extract of Ficus congensis

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    Silva Iwuanyanwu

    2011-04-01

    Full Text Available Ficus congensis (Moraceae is used traditionally in the treatment of various diseases including infectious diseases, infertility, and gastrointestinal disorders. Investigation of hexane extract of the stem bark using chromatographic techniques led to isolation of a xanthone, 1-hydroxy-3,7,8-trimethoxyxanthone (Decussatin. The compound was elucidated based on spectroscopic methods such as nuclear magnetic resonance (NMR, UV, IR, and mass spectrometry (MS. Decussatin and the hexane extract were screened in vitro for antibacterial and antifungal activities using broth microdilution (MHB and disc Agar diffusion (DAD techniques against Escheichia coli, Bacilus substilis, Klebsiela pneumonia, Staphylococcus aureus, Aspergillus fumigatus, Trichophyton mentagrophytes, Trichophyton rubrum and Candida albicans. Hexane extracts showed potent antibacterial activity against E. coli and B. subtilis with minimum inhibitory concentrations (MIC of 8 mg/mL and 5 mg/mL, respectively, while Decussatin of the highest concentration (8 mg/mL used in this study showed no appreciable antimicrobial activity. Only hexane extract was active against C. albicans with a MIC of 1 mg/mL.

  11. Synthesis and antimicrobial evaluation of new pyrano[4,3-b]pyran and Pyrano[3,2-c]chromene derivatives bearing a 2-thiophenoxyquinoline nucleus.

    Science.gov (United States)

    Makawana, Jigar A; Patel, Manish P; Patel, Ranjan G

    2012-04-01

    A new series of pyrano[4,3-b]pyran 4a-i and pyrano[3,2-c]chromene 6a-r derivatives bearing a 2-thiophenoxyquinoline nucleus were synthesized by reaction of 2-(4-(un)-substituted thiophenoxy)quinoline-3-carbaldehydes 2a-i with 6-methyl-4-hydroxypyran-2-one 3 and 4-hydroxy-6-(un)-substituted-2H-chromen-2-one 5a-b respectively and malononitrile at room temperature in the presence of KOH as a basic catalyst. All the compounds were screened against three Gram-positive bacteria (Streptococcus pneumoniae, Bacillus subtilis, Clostridium tetani), three Gram-negative bacteria (Salmonella typhi, Escherichia coli, Vibrio cholerae) and two fungi (Candida albicans, Aspergillus fumigatus) using the broth microdilution MIC (minimum inhibitory concentration) method. Upon antimicrobial screening, it was observed that the majority of the compounds were found to be active against Bacillus subtilis, Clostridium tetani and Candida albicans as compared to standard drugs. PMID:22105795

  12. In vitro Quinolones Susceptibility Analysis of Chinese Mycoplasma bovis Isolates and their Phylogenetic Scenarios based upon QRDRs of DNA Topoisomerases Revealing a Unique Transition in ParC

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    Riaz Mustafa1,2,3, Jingjing Qi1,2, Xiaoliang Ba1,2, Yingyu Chen1,4, Changmin Hu1,2, Xiaole Liu1,2, Lingling Tu5, Qingjie Peng5, Huanchun Chen1,2 and Aizhen Guo1,2*

    2013-07-01

    Full Text Available Mycoplasma bovis can cause different systemic problems in cattle, and recently has been resulted in huge economic losses in China. In vitro susceptibilities of 26 twice sub-cultured Chinese M. bovis field isolates were determined at physiological pH including PG45 through broth micro-dilution method. Except Huanggang isolate, all isolates and PG45 were in the sensitive range for levofloxacin, lomefloxacin and ciprofloxacin, whereas, for norfloxacin and nalidixic acid, they had shown intermediate resistant and complete resistant patterns, respectively. The multiple sequence analysis revealed point mutations in QRDRs of gyrA and parC genes of Huanggang isolate resulting in amino acid substitutions at positions 83 (S-F in GyrA (E. coli numbering and 80 (S-I in ParC proteins, the latter is reported for first time in M. bovis. Conclusively, fluoroquinolones are the potential veterinary therapeutic agents for mycoplasmosis in China and resistance to these agents comes through point mutations in QRDRs of gyrA and parC genes with ParC and GyrA mutation orientation.

  13. In vitro sensitivity of the amphibian pathogen Batrachochytrium dendrobatidis to antifungal therapeutics.

    Science.gov (United States)

    Woodward, A; Berger, L; Skerratt, L F

    2014-10-01

    Chytridiomycosis, a skin disease caused by Batrachochytrium dendrobatidis, has caused amphibian declines worldwide. Amphibians can be treated by percutaneous application of antimicrobials, but knowledge of in vitro susceptibility is lacking. Using a modified broth microdilution method, we describe the in vitro sensitivity of two Australian isolates of B. dendrobatidis to six antimicrobial agents. Growth inhibition was observed, by measurement of optical density, with all agents. Minimum inhibitory concentrations (µg/ml; isolate 1/2) were - voriconazole 0.016/0.008; itraconazole 0.032/0.016; terbinafine 0.063/0.063; fluconazole 0.31/0.31; chloramphenicol 12.5/12.5; amphotericin B 12.5/6.25. Killing effects on zoospores were assessed by observing motility. Amphotericin B and terbinafine killed zoospores within 5 and 30 min depending on concentration, but other antimicrobials were not effective at the highest concentrations tested (100 µg/ml). This knowledge will help in drug selection and treatment optimization. As terbinafine was potent and has rapid effects, study of its pharmacokinetics, safety and efficacy is recommended. PMID:25241392

  14. Antifungal Activity of Brazilian Propolis Microparticles against Yeasts Isolated from Vulvovaginal Candidiasis

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    Kelen Fátima Dalben Dota

    2011-01-01

    Full Text Available Propolis, a resinous compound produced by Apis mellifera L. bees, is known to possess a variety of biological activities and is applied in the therapy of various infectious diseases. The aim of this study was to evaluate the in vitro antifungal activity of propolis ethanol extract (PE and propolis microparticles (PMs obtained from a sample of Brazilian propolis against clinical yeast isolates of importance in the vulvovaginal candidiasis (VVC. PE was used to prepare the microparticles. Yeast isolates (n=89, obtained from vaginal exudates of patients with VVC, were exposed to the PE and the PMs. Moreover, the main antifungal drugs used in the treatment of VVC (Fluconazole, Voriconazole, Itraconazole, Ketoconazole, Miconazole and Amphotericin B were also tested. Minimum inhibitory concentration (MIC was determined according to the standard broth microdilution method. Some Candida albicans isolates showed resistance or dose-dependent susceptibility for the azolic drugs and Amphotericin B. Non-C. albicans isolates showed more resistance and dose-dependent susceptibility for the azolic drugs than C. albicans. However, all of them were sensitive or dose-dependent susceptible for Amphotericin B. All yeasts were inhibited by PE and PMs, with small variation, independent of the species of yeast. The overall results provided important information for the potential application of PMs in the therapy of VVC and the possible prevention of the occurrence of new symptomatic episodes.

  15. Antibacterial potential of the essential oil from Sideritis montana L. (Lamiaceae

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    Miladinović Dragoljub L.

    2012-01-01

    Full Text Available The antibacterial potential of essential oil from Serbian Sideritis montana L. (Lamiaceae on different bacteria was investigated. The essential oil was obtained from air dried aerial parts of the plant with 0.03% (w/w yield by hydrodistillation for 4 h using a Clevenger-type apparatus. The essential oil analyses were performed simultaneously by gas chromatography (GC and gas chromatography-mass spectrometry (GC-MS systems. Dominant compounds class is sesquiterpene hydrocarbons. The main constituents of oil were germacrene D (37,9% and trans-geraniol (26,1%. The antibacterial activity of the essential oil was tested towards 4 different bacteria, laboratory control strain obtained from the American Type Culture Collection (ATCC. Gram negative bacteria were represented by Escherichia coli ATCC 13706, Pseudomonas aeruginosa ATCC 27853, Salmonella enteritidis ATCC 13076 while researched Gram positive strain was Staphylococcus aureus ATCC 29213. Broth microdilution method was used to determine the minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC. Essential oil has been found to have antibacterial activity against all tested bacteria, with a MIC values ranging from 0.03 to 0.06 μl/ml and MBC values from 0.06 to 0.12 μl/ml. Reference antibiotic tetracycline was active in concentrations between 0.025 to 0.1 μg/ml. Essential oil from Serbian S. montana has high antibacterial potential which should be studied further.

  16. Effects of Myrcia ovata Cambess. essential oil on planktonic growth of gastrointestinal microorganisms and biofilm formation of Enterococcus faecalis

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    Cinthya S. Cândido

    2010-10-01

    Full Text Available The essential oil from the leaves of Myrcia ovata Cambess., commonly used in Brazil for the treatment of gastric illnesses, was screened for antimicrobial activity and action in the formation of microbial biofilms by Enterococcus faecalis. The oil was obtained by hydrodistillation using a clevenger-type system. Its chemical composition was analyzed using GC and GC-MS. Both MIC and MBC of the essential oil were determined by broth microdilution techniques and agar dilution method. The essential oil showed antimicrobial activity against E. faecalis, Escherichia coli, Pseudomonas aeruginosa, Salmonella choleraesuis, Staphylococcus aureus, Streptococcus pneumoniae and Candida parapsilosis. The results showed that the essential oil of M. ovata Cambess. was effective against the formation of biofilm by E. faecalis when compared with the control. Four volatile compounds, representing 92.1 % of the oil, were identified and geranial was the major component (50.4 %. At the best of our knowledge, this is the first report of the chemical composition and antimicrobial activity of the essential oil from leaves of M. ovata.

  17. Antimicrobial activity of Northwestern Mexican plants against Helicobacter pylori.

    Science.gov (United States)

    Robles-Zepeda, Ramón E; Velázquez-Contreras, Carlos A; Garibay-Escobar, Adriana; Gálvez-Ruiz, Juan C; Ruiz-Bustos, Eduardo

    2011-10-01

    Helicobacter pylori is the major etiologic agent of such gastric disorders as chronic active gastritis and gastric carcinoma. Over the past few years, the appearance of antibiotic-resistant bacteria has led to the development of better treatments, such as the use of natural products. This study evaluated the anti-H. pylori activity of 17 Mexican plants used mainly in the northwestern part of Mexico (Sonora) for the empirical treatment of gastrointestinal disorders. The anti-H. pylori activity of methanolic extracts of the plants was determined by using the broth microdilution method. The 50% minimum inhibitory concentrations ranged from less than 200 to 400 μg/mL for Castella tortuosa, Amphipterygium adstringens, Ibervillea sonorae, Pscalium decompositum, Krameria erecta, Selaginella lepidophylla, Pimpinella anisum, Marrubium vulgare, Ambrosia confertiflora, and Couterea latiflora and were greater than 800 μg/mL for Byophyllum pinnatum, Tecoma stans linnaeus, Kohleria deppena, Jatropha cuneata, Chenopodium ambrosoides, and Taxodium macronatum. Only Equisetum gigantum showed no activity against H. pylori. This study suggests the important role that these plants may have in the treatment of gastrointestinal disorders caused by H. pylori. The findings set the groundwork for further characterization and elucidation of the active compounds responsible for such activity. PMID:21663492

  18. In Vitro Antibacterial, Antifungal, Antibiofilm, Antioxidant, and Anticancer Properties of Isosteviol Isolated from Endangered Medicinal Plant Pittosporum tetraspermum

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    Naif Abdullah Al-Dhabi

    2015-01-01

    Full Text Available This study aimed to investigate the in vitro antibacterial, antifungal, antibiofilm, antioxidant, and anticancer properties of isosteviol isolated from endangered medicinal plant Pittosporum tetraspermum. Pure compound was obtained and characterized by column chromatography followed by 1H NMR, 13C NMR, IR, and mass spectral analysis. The antimicrobial activities of the compound were assessed by the broth microdilution method and the antioxidant properties were determined using reducing ability assay, DPPH scavenging assay, hydroxyl radical scavenging activity, and superoxide radical scavenging assay. Anticancer study was evaluated by following MTT assay. Column purification and spectrocopical analysis lead to identifying isosteviol from the crude ethyl acetate extract. The compound exhibited significant activity against bacteria such as Staphylococcus epidermidis (125 µg/mL, Staphylococcus aureus (125 µg/mL, and Klebsiella pneumoniae (62.5 µg/mL. The MIC of the compound against Candida albicans, Aspergillus niger, and Trichophyton mentagrophytes was 62.5, 125, and 500 µg/mL, respectively. The compound showed comparatively better antibiofilm activity against E. coli, S. typhi, and P. aeruginosa. Furthermore, it exhibited good antioxidant properties. Anticancer properties of the compound against Vero and MCF7 cell lines were its advantage. Novel isosteviol would be useful to reduce the infectious diseases caused by pathogenic microorganisms or slow the progress of various oxidative stress-related diseases.

  19. 抗菌肽对金黄色葡萄球菌抑制作用的研究%Study on the inhibitory effect of antimicrobial peptides-thanatin against staphylococcus aureus

    Institute of Scientific and Technical Information of China (English)

    刘付弟

    2011-01-01

    Objective To explore the inhibitory effect of antimicrobial peptide s-Thanatin against Staphylococcus aureus by broth microdilution. Methods The activity of s - Thanatin against Staphylococcus aureus was evaluated by the minimum inhibitory concentration (MIC). Results The results demonstrated that the MIC of s- Thanatin against Staphylococcus aureus was 50 mg/L. Conclusion A higher concentration of s- thanatin is necessary to inhibit common pathogenic Staphylococcus aureus. Therefore, the chemical structure of antibacterial peptide Thanatin should be modified to improve the activity against Staphylococcus aureus.%目的 探讨抗菌肽S-Thanatin对金黄色葡萄球菌的抑制效果.方法 采用肉汤微量稀释法研究抗菌肽S-Thanatin对金黄色葡萄球菌的最低抑菌浓度(MIC).结果 抗菌肽S-Thanatin对金黄色葡萄球菌MIC是50 mg/L.结论 抗菌肽S-Thanatin对临床常见的金黄色葡萄球菌需要较高浓度才有抑制作用,因此为进一步提高其抗金黄色葡萄球菌活性,需要对抗菌肽S-Thanatin的化学结构进行改进.

  20. The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

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    Marina Azevedo Souza

    2013-05-01

    Full Text Available Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazone and semicarbazone derivatives were also active against the pathogenic yeast Cryptococcus gattii (MICs of 0.10 and 0.20 µmol/mL, respectively. In addition, the lapachol thiosemicarbazone derivative was active against 11 clinical isolates of Paracoccidioides brasiliensis, with MICs ranging from 0.01-0.10 µmol/mL. The lapachol-derived thiosemicarbazone was not cytotoxic to normal cells at the concentrations that were active against fungi and bacteria. We synthesised, for the first time, thiosemicarbazone and semicarbazone derivatives of lapachol. The MICs for the lapachol-derived thiosemicarbazone against S. aureus, E. faecalis, C. gattii and several isolates of P. brasiliensis indicated that this compound has the potential to be developed into novel drugs to treat infections caused these microbes.

  1. Chemical Composition and In Vitro Antibacterial Activity of Mentha spicata Essential Oil against Common Food-Borne Pathogenic Bacteria.

    Science.gov (United States)

    Shahbazi, Yasser

    2015-01-01

    The aim of the present study was to investigate chemical composition and antibacterial activity of essential oil from the leaf of Mentha spicata plant against common food-borne pathogenic bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Listeria monocytogenes, Salmonella typhimurium, and Escherichia coli O157:H7). Chemical composition of the essential oil was identified by gas chromatography coupled with mass spectrometer detector (GC-MS). The antibacterial activity of the essential oil was evaluated by broth microdilution method and agar disk diffusion assay. According to the result of GC-MS analysis, 18 components were identified, accounting for 99.89% of the whole essential oil. The main components were carvone (78.76%), limonene (11.50%), β-bourbonene (11.23%), cis-dihydrocarveol (1.43%), trans-caryophyllene (1.04%), menthone (1.01%), menthol (1%), and terpinen-4-ol (0.99). The essential oil exhibited moderate level of antibacterial activity against all test microorganisms. In general, Gram-positive bacteria were more susceptible to M. spicata essential oil than Gram-negative bacteria. L. monocytogenes was the most sensitive of the microorganisms to the antibacterial activity of M. spicata essential oil (inhibition zone = 22 mm and MIC and MBC = 2.5 µL/mL). Based on our results, the essential oil of M. spicata plant collected from Kermanshah province, west of Iran, has a potential to be applied as antibacterial agent. PMID:26351584

  2. The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

    Science.gov (United States)

    Souza, Marina Azevêdo; Johann, Susana; Lima, Luciana Alves Rodrigues dos Santos; Campos, Fernanda Fraga; Mendes, Isolda Castro; Beraldo, Heloisa; de Souza-Fagundes, Elaine Maria; Cisalpino, Patrícia Silva; Rosa, Carlos Augusto; Alves, Tânia Maria de Almeida; de Sá, Nívea Pereira; Zani, Carlos Leomar

    2013-01-01

    Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs) of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazone and semicarbazone derivatives were also active against the pathogenic yeast Cryptococcus gattii (MICs of 0.10 and 0.20 µmol/mL, respectively). In addition, the lapachol thiosemicarbazone derivative was active against 11 clinical isolates of Paracoccidioides brasiliensis, with MICs ranging from 0.01-0.10 µmol/mL. The lapachol-derived thiosemicarbazone was not cytotoxic to normal cells at the concentrations that were active against fungi and bacteria. We synthesised, for the first time, thiosemicarbazone and semicarbazone derivatives of lapachol. The MICs for the lapachol-derived thiosemicarbazone against S. aureus, E. faecalis, C. gattii and several isolates of P. brasiliensis indicated that this compound has the potential to be developed into novel drugs to treat infections caused these microbes. PMID:23778660

  3. Synthesis and In Vitro Evaluation of New Thiosemicarbazone Derivatives as Potential Antimicrobial Agents

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    Zafer Asım Kaplancıklı

    2016-01-01

    Full Text Available In an effort to develop potent antimicrobial agents, new thiosemicarbazone derivatives were synthesized via the reaction of 4-[4-(trifluoromethylphenyl]thiosemicarbazide with aromatic aldehydes. The compounds were evaluated for their inhibitory effects on pathogenic bacteria and yeasts using the CLSI broth microdilution method. Microplate Alamar Blue Assay was also carried out to determine the antimycobacterial activities of the compounds against Mycobacterium tuberculosis H37Rv. Among these derivatives, compounds 5 and 11 were more effective against Enterococcus faecalis (ATCC 29212 than chloramphenicol, whereas compounds 1, 2, and 12 and chloramphenicol showed the same level of antibacterial activity against E. faecalis. Moreover, compound 2 and chloramphenicol exhibited the same level of antibacterial activity against Staphylococcus aureus. On the other hand, the most potent anticandidal derivatives were found as compounds 2 and 5. These derivatives and ketoconazole exhibited the same level of antifungal activity against Candida glabrata. According to the Microplate Alamar Blue Assay, the tested compounds showed weak to moderate antitubercular activity.

  4. Phytochemical Analysis and Antimicrobial, Antinociceptive, and Anti-Inflammatory Activities of Two Chemotypes of Pimenta pseudocaryophyllus (Myrtaceae).

    Science.gov (United States)

    de Paula, Joelma Abadia Marciano; Silva, Maria do Rosário Rodrigues; Costa, Maysa P; Diniz, Danielle Guimarães Almeida; Sá, Fabyola A S; Alves, Suzana Ferreira; Costa, Elson Alves; Lino, Roberta Campos; de Paula, José Realino

    2012-01-01

    Preparations from Pimenta pseudocaryophyllus (Gomes) L.R. Landrum (Myrtaceae) have been widely used in Brazilian folk medicine. This study aims to evaluate the antimicrobial activity of the crude ethanol extracts, fractions, semipurified substances, and essential oils obtained from leaves of two chemotypes of P. pseudocaryophyllus and to perform the antinociceptive and anti-inflammatory screening. The ethanol extracts were purified by column chromatography and main compounds were spectrally characterised (1D and 2D (1)H and (13)C NMR). The essential oils constituents were identified by GC/MS. The broth microdilution method was used for testing the antimicrobial activity. The abdominal contortions induced by acetic acid and the ear oedema induced by croton oil were used for screening of antinociceptive and anti-inflammatory activities, respectively. The phytochemical analysis resulted in the isolation of pentacyclic triterpenes, flavonoids, and phenol acids. The oleanolic acid showed the best profile of antibacterial activity for Gram-positive bacteria (31.2-125 μg mL(-1)), followed by the essential oil of the citral chemotype (62.5-250 μg mL(-1)). Among the semipurified substances, Ppm5, which contained gallic acid, was the most active for Candida spp. (31.2 μg mL(-1)) and Cryptococcus spp. (3.9-15.6 μg mL(-1)). The crude ethanol extract and fractions from citral chemotype showed antinociceptive and anti-inflammatory effects. PMID:23082081

  5. Phytochemical Analysis and Antimicrobial, Antinociceptive, and Anti-Inflammatory Activities of Two Chemotypes of Pimenta pseudocaryophyllus (Myrtaceae

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    Joelma Abadia Marciano de Paula

    2012-01-01

    Full Text Available Preparations from Pimenta pseudocaryophyllus (Gomes L.R. Landrum (Myrtaceae have been widely used in Brazilian folk medicine. This study aims to evaluate the antimicrobial activity of the crude ethanol extracts, fractions, semipurified substances, and essential oils obtained from leaves of two chemotypes of P. pseudocaryophyllus and to perform the antinociceptive and anti-inflammatory screening. The ethanol extracts were purified by column chromatography and main compounds were spectrally characterised (1D and 2D 1H and 13C NMR. The essential oils constituents were identified by GC/MS. The broth microdilution method was used for testing the antimicrobial activity. The abdominal contortions induced by acetic acid and the ear oedema induced by croton oil were used for screening of antinociceptive and anti-inflammatory activities, respectively. The phytochemical analysis resulted in the isolation of pentacyclic triterpenes, flavonoids, and phenol acids. The oleanolic acid showed the best profile of antibacterial activity for Gram-positive bacteria (31.2–125 μg mL−1, followed by the essential oil of the citral chemotype (62.5–250 μg mL−1. Among the semipurified substances, Ppm5, which contained gallic acid, was the most active for Candida spp. (31.2 μg mL−1 and Cryptococcus spp. (3.9–15.6 μg mL−1. The crude ethanol extract and fractions from citral chemotype showed antinociceptive and anti-inflammatory effects.

  6. Synthesis of Some N-Alkoxycarbonyl-N″-benzoyl-benzamidrazones(p-toluamidrazones) and 1,3,5-Trisubstituted 1,2,4-Triazole Derivatives from N-Benzoylimidates and their Antimicrobial and Anticancer Screening Studies

    Institute of Scientific and Technical Information of China (English)

    BEKIRCAN Olcay; KAHVECI Bahittin; OZGUMUS Osman Birol

    2007-01-01

    Some new N-alkoxycarbonyl-N″-benzoyl-benzamidrazones (p-toluamidrazones) 3a-3d,and 1,3,5-trisubstituted 1,2,4-triazole 4a-4h derivatives by starting from N-benzoylbenzimidates or N-benzoyl-p-toluimidates.The structures of compounds 3 and 4 were established on the basis of elemental analyses,IR,1H NMR,13C NMR and UV data.Antimicrobial experiments of the compounds performed by using agar-well diffusion and broth microdilution methods revealed that only compounds 3a-3d,4a and 4b showed inhibitory effect only on Candida albicans ATCC 60193.However,compound 4b had also specific antibacterial activity against Staphylococcus aureus ATCC 25923.The other compounds showed neither antifungal nor antibacterial activities.Compounds 3a,4a and 4b have been screened on three human tumor cell lines,breast cancer (MCF7),non small cell lung cancer (NCI-H460),and CNS cancer (SF-268) at the National Cancer Institute (NCI),USA,which were found to exhibit low antiproliferative activity.

  7. Antimicrobial Constituents and Effects of Blended Eucalyptus, Rosemary, Patchouli, Pine, and Cajuput Essential Oils.

    Science.gov (United States)

    Tadtong, Sarin; Puengseangdee, Chompunut; Prasertthanawut, Suwapit; Hongratanaworakit, Tapanee

    2016-02-01

    The synergistic antimicrobial activities of blended essential oil preparations composed of eucalyptus, rosemary, patchouli, pine and cajuput oils were evaluated against various pathogenic microorganisms. They exhibited antimicrobial activity in the agar disc diffusion assay against the Gram-positive bacteria, Staphylococcus aureus and S. epidenidis, the Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, and the yeast, Candida albicans. The minimum inhibitory concentration (MIC) of these preparations was evaluated by the broth microdilution method. We found that the best synergistic antibacterial preparation (the sum of fractional inhibitory concentrations, Σfic rosemary, and mineral oils (volume ratio 4:4:2) without patchouli, pine or cajuput oils added, while most preparations showed an antagonistic anticandidal effect (Σfic > 1). The blended essential oil preparations were characterized for their components by GC/MS, and contained 1,8-cineole as a major component. However, the preparation containing the highest amount of 1,8-cineole did not exhibit the best synergistic effect, which was shown by the preparation comprising the highest amount of α-pinene. Hence, we concluded that the differential antibacterial effect of either blended oil preparations or single/pure essential oils was influenced by the amount of α-pinene and the number of active components in either the blended preparations or single/pure essential oils. PMID:27032218

  8. Antibacterial efficacy of Mangifera indica L. kernel and Ocimum sanctum L. leaves against Enterococcus faecalis dentinal biofilm

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    Arunajatesan Subbiya

    2013-01-01

    Full Text Available Introduction: The Enterococcus faecalis biofilm in the root canal makes it difficult to be eradicated by the conventional irrigants with no toxicity to the tissues. Hence, plant products with least side effects are explored for their use as irrigants in the root canal therapy. Aim: To evaluate and compare the antibacterial efficacy of Mangifera indica L. kernel (mango kernel and Ocimum sanctum L. leaves (tulsi extracts with conventional irrigants (5% sodium hypochlorite (NaOCl and 2% chlorhexidine against E. faecalis dentinal biofilm. Materials and Methods: Agar diffusion and broth microdilution assay was performed with the herbal extracts and conventional irrigants (2% chlorhexidine and 5% NaOCl against E. faecalis planktonic cells. The assay was extended onto 3 week E. faecalis dentinal biofilm. Results: Significant reduction of colony forming units (CFU/mL was observed for the herbal groups and the antibacterial activity of the herbal groups was at par with 5% NaOCl. Conclusions: The antibacterial activity of these herbal extracts is found to be comparable with that of conventional irrigants both on the biofilm and planktonic counterparts.

  9. In vitro and in vivo antifungal activity of natural inhibitors against Penicillium expansum

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    Claudia Fieira

    2013-02-01

    Full Text Available Penicillium expansum is the causative agent of apple blue mold. The inhibitory effects of the capsaicin derived from Capsicum spp. fruits and yeast Hansenula wingei against P. expansum were evaluated in an in vitro and in in vivo assay using Fuji apples. The minimum inhibitory concentration of capsaicin determined using the broth micro-dilution method was 122.16 µg mL-1. Capsaicin did not reduce blue mold incidence in apples. However, it was able to delay fungal growth in the first 14 days of the in vivo assay. The in vivo effect of the yeast Hansenula wingei AM2(-2, alone and combined with thiabendazole at low dosage (40 µg mL-1, on the incidence of apple diseases caused by P. expansum was also described. H. wingei AM2(-2 combined with a low fungicide dosage (10% of the dosage recommended by the manufacturer showed the best efficacy (100% up to 7 days of storage at 21 ºC, later showing a non-statistically different decrease (p > 0.05 after 14 (80.45% and 21 days (72.13%, respectively. These results contribute providing new options for using antifungal agents against Penicillium expansum.

  10. Antimicrobial activity of Rosmarinus officinalis against oral pathogens: relevance of carnosic acid and carnosol.

    Science.gov (United States)

    Bernardes, Wagner A; Lucarini, Rodrigo; Tozatti, Marcos G; Souza, Maria G M; Silva, Márcio L Andrade; Filho, Ademar A da Silva; Martins, Carlos H Gomes; Crotti, Antonio E Miller; Pauletti, Patricia M; Groppo, Milton; Cunha, Wilson R

    2010-07-01

    The in vitro inhibitory activity of crude EtOH/H2O extracts from the leaves and stems of Rosmarinus officinalis L. was evaluated against the following microorganisms responsible for initiating dental caries: Streptococcus mutans, S. salivarius, S. sobrinus, S. mitis, S. sanguinis, and Enterococcus faecalis. Minimum inhibitory concentrations (MIC) were determined with the broth microdilution method. The bioassay-guided fractionation of the leaf extract, which displayed the higher antibacterial activity than the stem extract, led to the identification of carnosic acid (2) and carnosol (3) as the major compounds in the fraction displaying the highest activity, as identified by HPLC analysis. Rosmarinic acid (1), detected in another fraction, did not display any activity against the selected microorganisms. HPLC Analysis revealed the presence of low amounts of ursolic acid (4) and oleanolic acid (5) in the obtained fractions. The results suggest that the antimicrobial activity of the extract from the leaves of R. officinalis may be ascribed mainly to the action of 2 and 3. PMID:20658673

  11. Minocycline activity tested against Acinetobacter baumannii complex, Stenotrophomonas maltophilia, and Burkholderia cepacia species complex isolates from a global surveillance program (2013).

    Science.gov (United States)

    Flamm, Robert K; Castanheira, Mariana; Streit, Jennifer M; Jones, Ronald N

    2016-07-01

    Clinical isolates of Acinetobacter baumannii complex (1312), Stenotrophomonas maltophilia (464), and Burkholderia cepacia species complex (30) were selected from medical centers in the United States (USA), Europe and the Mediterranean (EU-M) region, Latin America, and Asia Pacific. Only one isolate per infected patient episode was included and local identifications were confirmed by the monitoring laboratory. Susceptibility testing was performed at the monitoring laboratory using the reference broth microdilution method as described by Clinical and Laboratory Standards Institute (CLSI). A. baumannii complex were classified as MDR (multi-drug resistant [MDR]; nonsusceptible to ≥1 agent in ≥3 antimicrobial classes) and extensively drug-resistant (XDR; nonsusceptible to ≥1 agent in all but ≤2 antimicrobial classes). A total of 81.6% of A. baumannii complex were MDR. Colistin was the most active agent against MDR A. baumannii complex. Minocycline was the most active "tetracycline" against these organisms based on susceptibility. Against B. cepacia, trimethoprim-sulfamethoxazole (MIC90, 2 μg/mL; 100.0% susceptible) was the most active agent tested. Overall, minocycline was the most active tetracycline tested against A. baumannii complex and S. maltophilia isolates collected from patients throughout EU-M, USA, Latin America, and the Asia-Pacific. Minocycline, particularly the intravenous formulation, has activity against several ESKAPE pathogens and merits consideration in seriously ill patients where treatment options may be limited due to the presence of MDR bacteria. PMID:27112832

  12. Potentiation activity of multiple antibacterial agents by Salvianolate from the Chinese medicine Danshen against methicillin-resistant Staphylococcus aureus (MRSA).

    Science.gov (United States)

    Liu, Qing-Qing; Han, Jun; Zuo, Guo-Ying; Wang, Gen-Chun; Tang, Hua-Shu

    2016-05-01

    Salvianolate (SAL) is a prescribed medicine from the Chinese herb Danshen (Salvia miltiorrhiza Bunge). It has been widely used in treatment of coronary and other diseases with significant effects. The in vitro antimicrobial activities of SAL against infectious pathogens were assayed and its combined effects on 10 clinical isolates of SCCmec III type methicillin-resistant Staphylococcus aureus (MRSA) with ten antibiotics were evaluated. Susceptibility to each agent alone was tested using a broth microdilution method, and the chequerboard and time-kill experiments were used for the combined activities. The results showed MIC was 128-256 mg/L for SAL used alone against MRSA. Significant synergies were observed for SAL/Ampicillin (Fosfomycin, Erythromycin, Piperacillin-tazobactam or Clindamycin) combination against over half of the isolates, with their MICs reduced by times of dilution (TOD) to 4-32 (FICIs 0.375-0.5), respectively. SAL/AMP combination showed the best combined effect of synergy on bacteriostatic and bactericidal activities, while SAL/AMK combination reversed the resistance of MRSA to AMK. The results demonstrated that SAL enhanced widely the in vitro anti-MRSA efficacy of the ten antibacterial agents, which had potential for combinatory therapy of patients infected with MRSA and warrants further investigations. PMID:26639445

  13. Antifungal activity of raw extract and flavanons isolated from Piper ecuadorense from Ecuador

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    Jorge Ramirez

    2013-04-01

    Full Text Available The MeOH extract of Piper ecuadorense Sodiro, Piperaceae, was chosen for metabolite isolation and elucidation due to the strong antifungal activity exhibited, measured by means of the broth microdilution method. Two known flavonoids: pinostrobin (1 and pinocembrin (2 were isolated from 4.16 g. of dichloromethane extract by column chromatography, using a gradient of hexane/EtOAc. A total of 20 mg of 1 were obtained from the fraction eluted with hexane-EtOAc 95:5 v/v, and 100 mg of 2 were obtained from the fraction eluted with hexane-EtOAc 85:15 v/v. The MIC values of the MeOH extract was 31.25 µg/mL for Trichophyton mentagrophytes ATCC® 28185 and 62.5 µg/mL for Trichophyton rubrum ATCC® 28188. The MIC value of pinocembrin was 125 µg/mL for Trichophyton mentagrophytes ATCC® 28185 and Trichophyton rubrum ATCC® 28188. Pinostrobin in antifungal test was not active against fungi tested.

  14. Antifungal susceptibility testing of Candida in the Clinical Laboratory: how to do it, when to do it, and how to interpret it

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    Esther Manso

    2014-06-01

    Full Text Available Significant changes in the management of fungaemia have occurred in the last decade with increased use of fluconazole prophylaxis, of empirical treatment and of echinocandins as first-line agents for documented disease. The emergence of drug resistance in fungal pathogens has a profound impact on human health given limited number of antifungal drugs. Antifungal resistance in Candida may be either intrinsic or acquired and may be encountered in the antifungal drug exposed but also the antifungal drug naïve patient The variation in resistance rates between centers emphasizes that it is essential to have knowledge of the local Candida species distribution and antifungal resistance rates to guide initial therapy for Candida BSI. Moreover, all Candida isolates from blood and normally sterile sites should be identified to the species level. The Clinical and Laboratory Standards Institute and European Committee on Antimicrobial Susceptibility Testing have developed breakpoints and epidemiological cutoff values that are now established for Candida spp. Clinical microbiology laboratories will be employed commercial susceptibility assays, rather than reference broth microdilution methods and comparative studies are particularly important. Vitek 2®, Etest® and Sensititre YeastOne® provided a high degree of essential agreement and comparable sensitivity and specificity to BMD-RPMI for identifying resistance to azole and echinocandins in Candida spp.

  15. Echinocandin resistance and population structure of invasive Candida glabrata isolates from two university hospitals in Germany and Austria.

    Science.gov (United States)

    Klotz, Ulrike; Schmidt, Dirk; Willinger, Birgit; Steinmann, Eike; Buer, Jan; Rath, Peter-Michael; Steinmann, Joerg

    2016-05-01

    Echinocandin resistance in Candida glabrata is emerging and is associated with the presence of FKS mutations. In this study, we analysed the antifungal susceptibility, presence of FKS mutations and clonality of C. glabrata blood culture isolates from two hospitals in Germany and Austria. Susceptibility testing of 64 C. glabrata bloodstream isolates from two university hospitals was performed with broth microdilution method according to EUCAST. In addition, all isolates were screened for FKS mutations. Molecular fingerprinting was performed by microsatellite PCR with three separate primer pairs and semiautomated repetitive sequenced-based PCR (rep-PCR). One C. glabrata isolate from Germany (1.5%) was echinocandin resistant, with a corresponding mutation in FKS2 gene hot spot 1. The discriminatory power of microsatellite PCR was higher than that of rep-PCR (Simpson Index of 0.94 vs. 0.88); microsatellite PCR created 31 separate genotypes, whereas rep-PCR created 17. Predominant genotypes or clusters of isolates from Germany and Austria were present, with no epidemiological evidence of nosocomial transmissions. Although we found a low incidence of echinocandin resistance in C. glabrata in our settings, further surveillance projects in central Europe are warranted for monitoring future epidemiological trends. The genetic population structure of C. glabrata demonstrates overrepresented geographical clusters. PMID:26806376

  16. Antimicrobial susceptibility of Mycoplasma bovis isolates from veal calves and dairy cattle in the Netherlands.

    Science.gov (United States)

    Heuvelink, Annet; Reugebrink, Constance; Mars, Jet

    2016-06-30

    Control of Mycoplasma bovis infections depends on good husbandry practices and antibiotic treatment. To allow more prudent use of antimicrobial drugs, there is a need for information on the susceptibility profile of this pathogen. The objective of the present study was to analyse the in vitro antimicrobial susceptibility of clinical M. bovis isolates in the Netherlands. The collection comprised 95 bovine isolates, originating from lungs (n=56), mastitis milk (n=27), and synovial fluid (n=12), collected between 2008 and 2014. Minimal inhibitory concentrations (MICs) were assessed by broth microdilution, both by using in-house prepared MIC plates and by using commercially available MIC plates. For each antimicrobial agent, the range of MIC results, the MIC50, and MIC90 values were calculated. M. bovis strains recently isolated in the Netherlands appeared to be characterized by relatively high MIC values for antimicrobial agents that, until now, have been recommended by the Dutch Association of Veterinarians for treating pneumonia caused by Mycoplasma species. Fluoroquinolones appeared to be the most efficacious in inhibiting M. bovis growth, followed by tulathromycin and oxytetracycline. The highest MIC values were obtained for erythromycin, tilmicosin, and tylosin. Future studies should be done on determining M. bovis specific clinical breakpoints, standardization of methods to determine MIC values as well as molecular studies on detection of antimicrobial resistance mechanisms of M. bovis isolates to develop PCR assays for determining resistance. PMID:27259820

  17. Diverse Genetic Background of Multidrug-Resistant Pseudomonas aeruginosa from Mainland China, and Emergence of an Extensively Drug-Resistant ST292 Clone in Kunming.

    Science.gov (United States)

    Fan, Xin; Wu, Yue; Xiao, Meng; Xu, Zhi-Peng; Kudinha, Timothy; Bazaj, Alda; Kong, Fanrong; Xu, Ying-Chun

    2016-01-01

    For a better understanding of the multidrug resistant Pseudomonas aeruginosa (MDR-PA) epidemiology in mainland China, a nationwide surveillance network of 27 tertiary hospitals was established. Non-duplicate MDR-PA isolates from 254 cases of nosocomial infections, were collected during the period August 2011 to July 2012. Minimum inhibitory concentrations (MICs) of nine antimicrobial agents were determined by broth micro-dilution method according to the CLSI guidelines [M7-A10]. Genotyping analysis was performed by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). The presence of acquired carbapenemases was also determined by molecular approaches for 233 carbapenem-resistant isolates. Carbapenemase genes were detected in 19 (8.2%) isolates, with 13 of these isolates encoding IMP-type enzymes, five with VIM-2, and one with KPC-2. MLST analysis revealed significant genetic diversity among the MDR-PA isolates studied, and 91 STs (including 17 novel STs) were identified. However, a long-term outbreak of an emerging extensively drug-resistant (XDR) ST292/PFGE genotype A clone was detected in a hospital from Southwest China. This study has demonstrated that MDR-PA in mainland China have evolved from diverse genetic backgrounds. Evidence of clonal dissemination of the organism and nosocomial outbreaks in some regions, suggest a need to strengthen existing infection control measures. PMID:27198004

  18. Therapeutic switching: from antidermatophytic essential oils to new leishmanicidal products

    Directory of Open Access Journals (Sweden)

    Emeline Houël

    2015-02-01

    Full Text Available This study examined whether the antidermatophytic activity of essential oils (EOs can be used as an indicator for the discovery of active natural products against Leishmania amazonensis. The aerial parts of seven plants were hydrodistilled. Using broth microdilution techniques, the obtained EOs were tested against three strains of dermatophytes (Trichophyton mentagrophytes, Microsporum gypseum and Microsporum canis. To compare the EOs antifungal and antiparasitic effects, the EOs activities against axenic amastigotes of L. amazonensis were concurrently evaluated. For the most promising EOs, their antileishmanial activities against parasites infecting peritoneal macrophages of BALB/c mice were measured. The most interesting antifungal candidates were the EOs from Cymbopogon citratus, Otacanthus azureus and Protium heptaphyllum, whereas O. azureus, Piper hispidum and P. heptaphyllum EOs exhibited the lowest 50% inhibitory concentration (IC50 values against axenic amastigotes, thus revealing a certain correspondence between both activities. The P. hispidum EO was identified as the most promising product in the results from the infected macrophages model (IC50: 4.7 µg/mL, safety index: 8. The most abundant compounds found in this EO were sesquiterpenes, notably curzerene and furanodiene. Eventually, the evaluation of the antidermatophytic activity of EOs appears to be an efficient method for identifying new potential drugs for the treatment of L. amazonensis.

  19. Antimicrobial Susceptibility and Genetic Characterisation of Burkholderia pseudomallei Isolated from Malaysian Patients

    Directory of Open Access Journals (Sweden)

    Yalda Khosravi

    2014-01-01

    Full Text Available Burkholderia pseudomallei, the causative agent of melioidosis, is intrinsically resistant to many antibiotics. Ceftazidime (CAZ, the synthetic β-lactam, is normally used as the first-line antibiotic therapy for treatment of melioidosis. However, acquired CAZ resistance can develop in vivo during treatment with CAZ, leading to mortality if therapy is not switched to a different antibiotic(s in a timely manner. In this study, susceptibilities of 81 B. pseudomallei isolates to nine different antimicrobial agents were determined using the disk diffusion method, broth microdilution test and Etest. Highest percentage of susceptibility was demonstrated to CAZ, amoxicillin/clavulanic acid, meropenem, imipenem, and trimethoprim/sulfamethoxazole. Although these drugs demonstrated the highest percentage of susceptibility in B. pseudomallei, the overall results underline the importance of the emergence of resistance in this organism. PCR results showed that, of the 81 B. pseudomallei, six multidrug resistant (MDR isolates carried bpeB, amrB, and BPSS1119 and penA genes. Genotyping of the isolates using random amplified polymorphic DNA analysis showed six different PCR fingerprinting patterns generated from the six MDR isolates clusters (A and eight PCR fingerprinting patterns generated for the remaining 75 non-MDR isolates clusters (B.

  20. Actividad antifúngica de extractos de biomasa celular de neem sobre aislamientos de dermatofitos

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    Daniel Iván Ospina Salazar

    2014-12-01

    Full Text Available Cell lines of neem (Azadirachta indica A. Juss. grown in liquid suspension have shown to produce bioactive secondary metabolites, particularly triterpenoids. In consequence, its use as a control of medical microorganisms (like dermatophytes is proposed. The main goal of this study was to assess the antifungal activity of methanolic extracts from neem cultured cell suspensions on several isolates of Trichophyton mentagrophytes (five isolates, Trichophyton rubrum (five isolates and Epidermophyton floccosum (three isolates. Neem cell suspension cultures were scaled up, from which a raw methanolic extract was obtained. This extract was fractionated by silica gel column chromatography. The raw methanolic extract and its fractions were used in order to determine the Minimal Inhibitory Concentration (MIC on the dermatophytes isolates by following M38-A2 broth microdilution method. Antimycotic Terbinafine was used as positive control. The results shown that neem raw cellular biomass extract inhibits the growth of T. mentagrophytes, T. rubrum and E. floccosum in at least 100%. In the evaluations of the separated fractions, it was observed that the low polarity fractions had higher antifungal activity (MIC=109 μg/mL than the raw extract per se (MIC=2500 μg/mL and the most polar ones (MIC=7000 μg/mL. The latter suggest that neem cells cultured in liquid suspension produces compounds with antifungal activity, being more active those present in the low polarity fractions.