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Sample records for agar dilution methods

  1. Screening fungicides for use in fish culture: Evaluation of the agar plug transfer, cellophane transfer, and agar dilution methods

    USGS Publications Warehouse

    Bailey, Tom A.

    1983-01-01

    The reliability, reproducibility, and usefulness of three screening methods -- the cellophane transfer, the agar plug transfer, and the agar dilution -- to screen aquatic fungicides were evaluated. Achlya flagellata and Saprolegnia hypogyna were exposed to 1, 10, and 100 mg/L of malachite green to test each method. The cellophane transfer and agar plug transfer techniques had similar reliability and reproducibility in rating fungicidal activity, and were both superior to the agar dilution technique. The agar plug transfer and agar dilution techniques adequately projected in vivo activity of malachite green, but the cellophane transfer technique overestimated its activity. Overall, the agar plug transfer technique most accurately rated the activity of malachite green and was the easiest test to perform. It therefore appears to be the method of choice for testing aquatic fungicides.

  2. Evaluation of agar dilution and broth microdilution methods to determine the disinfectant susceptibility.

    PubMed

    Wu, Guoyan; Yang, Qianru; Long, Mei; Guo, Lijuan; Li, Bei; Meng, Yue; Zhang, Anyun; Wang, Hongning; Liu, Shuliang; Zou, Likou

    2015-11-01

    A variety of disinfectants have been widely used in veterinary hygiene, food industries and environments, which could induce the development of bacterial resistance to disinfectants. The methods used to investigate antimicrobial effects of disinfectant vary considerably among studies, making comparisons difficult. In this study, agar dilution and broth microdilution methods were used to compare the antimicrobial activities of four quaternary ammonium compounds (QACs) against foodborne and zoonotic pathogens. The potential relationship between the presence of QACs resistance genes and phenotypic resistance to QACs was also investigated. Our results indicated that the minimum inhibitory concentrations (MICs) determined by two methods might be different depended upon different QACs and bacteria applied. Regardless of the testing methods, Klebsiella pneumoniae was more tolerant among Gram-negative strains to four QACs, followed by Salmonella and Escherichia coli. The agreement between MICs obtained by the two methods was good, for benzalkonium chloride (78.15%), didecyldimethylammonium chloride (DDAC) (82.35%), cetylpyridinium chloride (CTPC) (97.48%) and cetyltrimethylammonium bromide (CTAB) (99.16%), respectively. Among all Gram-negative bacteria, 94.55% (n=52) of qacEΔ1-positive strains showed higher MICs (512 mg l(-1)) to CTAB. The qacEΔ1 gene was highly associated (P<0.05) with the high MICs of QACs (⩾512 mg l(-1)). In addition, DDAC remained as the most effective disinfectant against both Gram-positive and Gram-negative bacteria. This is the first study that compared the agar dilution and broth microdilution methods to assess the antimicrobial activity of QACs. The study demonstrated the need to standardize method that would be used in evaluating QACs antimicrobial properties in the future.

  3. Comparison of E-test with agar dilution methods in testing susceptibility of N. gonorrhoeae to azithromycin.

    PubMed

    Yasin, R M; Suan, K A; Meng, C Y

    1997-05-01

    A single dose of a new antibiotic, azithromycin, has been shown to be effective in the treatment of uncomplicated Neisseria gonorrhoeae. A clinical study was conducted to assess the in vitro susceptibility of N gonorrhoeae to azithromycin and compare the reliability of results obtained using the new E-test methodology for determination of the minimum inhibitory concentration (MIC) of antibiotic with those obtained through the standard agar dilution method. 135 clinical isolates of N gonorrhoeae were obtained from patients attending hospital-based sexually transmitted disease clinics in five geographic locations in Malaysia. 76 of the isolates were penicillinase-producing N gonorrhoeae and 69 were high-level tetracycline-resistant N gonorrhoeae. All isolates were susceptible to azithromycin based on the susceptible MIC breakpoint of 2.0 mcg/ml. The MICs ranged from 0.0078-0.25 mcg/ml by agar dilution method and from 0.016-0.50 mcg/ml by E-test. Agreement between these two methods was 97.8%. The single-dose regime and good antigonococcal and antichlamydial activity of azithromycin make this antibiotic a suitable treatment choice. Moreover, the findings of this study suggest that the simpler, faster E-test is as reliable as the agar dilution method. Given the tendency of the antimicrobial susceptibility pattern of N gonorrhoeae to change rapidly, it is important to monitor MICs to detect the emergence of resistance.

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

    PubMed

    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.

  5. Assessment of Etest as an alternative to agar dilution for antimicrobial susceptibility testing of Neisseria gonorrhoeae.

    PubMed

    Liu, Hsi; Taylor, Thomas H; Pettus, Kevin; Trees, David

    2014-05-01

    We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory Improvement Amendments (CLIA)-approved CLSI standard agar dilution method and, separately, by the Etest according to the manufacturer's recommendations. The MICs were determined and compared. Ten laboratory-generated mutants were used to simulate substantially nonsusceptible specimens. The Etest and agar dilution methods were well correlated. Statistical tests produced regression R2 values of 88%, 82%, and 85% and Pearson correlation coefficients of 92%, 91%, and 92% for ceftriaxone, cefixime, and cefpodoxime, respectively. When paired comparisons were made, the two tests were 88.7%, 80%, and 87% within 1 log2 dilution from each other for ceftriaxone, cefixime, and cefpodoxime, respectively. The within-2-log2 agreements were 99.1%, 98.3%, and 94.8% for ceftriaxone, cefixime, and cefpodoxime, respectively. Notwithstanding the good correlations and the within-2-log2 general agreement, the Etest results produced slightly lower MICs than the agar dilution results. In conclusion, we found that the Etest can be effectively used as an alternative to agar dilution testing to determine the susceptibility of N. gonorrhoeae to ceftriaxone, cefixime, and cefpodoxime, although we recommend further research into extremely resistant isolates. For isolates within the typical range of clinical MICs, reexamination of the Etest interpretation of susceptible and nonsusceptible categories would likely allow for successful transition from agar dilution to the Etest.

  6. 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.

    PubMed

    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

    2007-10-01

    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 from patient materials during a selective digestive decontamination or selective oral decontamination trial in an intensive-care unit. Disk diffusion is an unreliable method to measure susceptibility to colistin. High error rates and low levels of reproducibility were observed in the disk diffusion test. The colistin Etest, agar dilution, and the VITEK 2 showed a high level of agreement with the broth microdilution reference method. Heteroresistance for colistin was observed in six Enterobacter cloacae isolates and in one Acinetobacter baumannii isolate. This is the first report of heteroresistance to colistin in E. cloacae isolates. Resistance to colistin in these isolates seemed to be induced upon exposure to colistin rather than being caused by stable mutations. Heteroresistant isolates could be detected in the broth microdilution, agar dilution, Etest, or disk diffusion test. The VITEK 2 displayed low sensitivity in the detection of heteroresistant subpopulations of E. cloacae. The VITEK 2 colistin susceptibility test can therefore be considered to be a reliable tool to determine susceptibility to colistin in isolates of genera that are known not to exhibit resistant subpopulations. In isolates of genera known to (occasionally) exhibit heteroresistance, an alternative susceptibility testing method capable of detecting heteroresistance should be used.

  7. Tetrazolium reduction as an aid for streptococcal growth detection with agar dilution susceptibility testing.

    PubMed Central

    Coudron, P E; Ford, J M; Dalton, H P

    1983-01-01

    A dye reduction method for determining a definitive endpoint with agar dilution susceptibility testing has been developed. Bacterial growth was determined by applying to the inoculum spot a dye solution containing 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride and phenazine methosulfate. Viable colonies reduced the tetrazolium salt to a visible red color within 3 to 5 min. The minimum inhibitory concentrations of six antimicrobial agents tested against 167 clinical streptococcal isolates were recorded before and after the addition of the tetrazolium-phenazine methosulfate solution. A total of 252 discrepancies (25%) were observed, and of these, 30 (12%) differed by more than one tested antibiotic concentration. Endpoint reproducibility of the dye procedure was assessed by four technologists in a double-blind study. A 2.7-fold reduction in disagreement was observed when the dye was used. Use of the tetrazolium-phenazine methosulfate solution involves little deviation from standard antimicrobial susceptibility test procedures and yields more accurate, as well as reproducible, susceptibility results. PMID:6630459

  8. Drug susceptibility testing by dilution methods.

    PubMed

    Jeannot, Katy; Plésiat, Patrick

    2014-01-01

    Serial twofold dilution methods are widely used to assess the bacteriostatic activities of antibiotics. This can be achieved by dilution of considered drugs in agar medium or in culture broth, and inoculation by calibrated inoculums. Although seemingly simple, these methods are greatly influenced by the experimental conditions used and may lead to discrepant results, in particular with untrained investigators. The present step-by-step protocol has been validated for Pseudomonas species, including P. aeruginosa. Introduction of appropriate control strains is crucial to ascertain minimal inhibitory concentration values and compare the results of independent experiments.

  9. Collaborative investigation of broth microdilution and semisolid agar dilution for in vitro susceptibility testing of Candida albicans.

    PubMed

    Shawar, R; Paetznick, V; Witte, Z; Ensign, L G; Anaissie, E; LaRocco, M

    1992-08-01

    A study was performed in two laboratories to evaluate the effect of growth medium and test methodology on inter- and intralaboratory variations in the MICs of amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITRA), and the triazole Sch 39304 (SCH) against 14 isolates of Candida albicans. Testing was performed by broth microdilution and semisolid agar dilution with the following media, buffered to pH 7.0 with morpholinepropanesulfonic acid (MOPS): buffered yeast nitrogen base (BYNB), Eagle's minimal essential medium (EMEM), RPMI 1640 medium (RPMI), and synthetic amino acid medium for fungi (SAAMF). Inocula were standardized spectrophotometrically, and endpoints were defined by the complete absence of growth for AMB and by no more than 25% of the growth in the drug-free control for all other agents. Comparative analyses of median MICs, as determined by each test method, were made for all drug-medium combinations. Both methods yielded similar (+/- 1 twofold dilution) median MICs for AMB in EMEM and RPMI, 5FC in all media, and FLU in EMEM, RPMI, and SAAMF. In contrast, substantial between-method variations in median MICs were seen for AMB in BYNB and SAAMF, FLU In BYNB, and ITRA and SCH in all media. Interlaboratory concordance of median MICs was good for AMB, 5FC, and FLU but poor for ITRA and SCH in all media. Endpoint determinations were analyzed by use of kappa statistical analyses for evaluating the strength of observer agreement. Moderate to almost perfect interlaboratory agreement occurred with AMB and 5FC in all media and with FLU in EMEM, RPMI, and SAAMF, irrespective of the test method. Slight to almost perfect interlaboratory agreement occurred with ITRA and SCH in EMEM, RPMI, and SAAMF when tested by semisolid agar dilution but not broth microdilution. Kappa values assessing intralaboratory agreement between methods were high for 5FC in all media, for AMB in BYNB, ENEM, and RPMI, and for FLU in EMEM, RPMI, and SAAMF. One laboratory

  10. Collaborative investigation of broth microdilution and semisolid agar dilution for in vitro susceptibility testing of Candida albicans.

    PubMed Central

    Shawar, R; Paetznick, V; Witte, Z; Ensign, L G; Anaissie, E; LaRocco, M

    1992-01-01

    A study was performed in two laboratories to evaluate the effect of growth medium and test methodology on inter- and intralaboratory variations in the MICs of amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITRA), and the triazole Sch 39304 (SCH) against 14 isolates of Candida albicans. Testing was performed by broth microdilution and semisolid agar dilution with the following media, buffered to pH 7.0 with morpholinepropanesulfonic acid (MOPS): buffered yeast nitrogen base (BYNB), Eagle's minimal essential medium (EMEM), RPMI 1640 medium (RPMI), and synthetic amino acid medium for fungi (SAAMF). Inocula were standardized spectrophotometrically, and endpoints were defined by the complete absence of growth for AMB and by no more than 25% of the growth in the drug-free control for all other agents. Comparative analyses of median MICs, as determined by each test method, were made for all drug-medium combinations. Both methods yielded similar (+/- 1 twofold dilution) median MICs for AMB in EMEM and RPMI, 5FC in all media, and FLU in EMEM, RPMI, and SAAMF. In contrast, substantial between-method variations in median MICs were seen for AMB in BYNB and SAAMF, FLU In BYNB, and ITRA and SCH in all media. Interlaboratory concordance of median MICs was good for AMB, 5FC, and FLU but poor for ITRA and SCH in all media. Endpoint determinations were analyzed by use of kappa statistical analyses for evaluating the strength of observer agreement. Moderate to almost perfect interlaboratory agreement occurred with AMB and 5FC in all media and with FLU in EMEM, RPMI, and SAAMF, irrespective of the test method. Slight to almost perfect interlaboratory agreement occurred with ITRA and SCH in EMEM, RPMI, and SAAMF when tested by semisolid agar dilution but not broth microdilution. Kappa values assessing intralaboratory agreement between methods were high for 5FC in all media, for AMB in BYNB, ENEM, and RPMI, and for FLU in EMEM, RPMI, and SAAMF. One laboratory

  11. Collaborative investigation of broth microdilution and semisolid agar dilution for in vitro susceptibility testing of Candida albicans.

    PubMed

    Shawar, R; Paetznick, V; Witte, Z; Ensign, L G; Anaissie, E; LaRocco, M

    1992-08-01

    A study was performed in two laboratories to evaluate the effect of growth medium and test methodology on inter- and intralaboratory variations in the MICs of amphotericin B (AMB), flucytosine (5FC), fluconazole (FLU), itraconazole (ITRA), and the triazole Sch 39304 (SCH) against 14 isolates of Candida albicans. Testing was performed by broth microdilution and semisolid agar dilution with the following media, buffered to pH 7.0 with morpholinepropanesulfonic acid (MOPS): buffered yeast nitrogen base (BYNB), Eagle's minimal essential medium (EMEM), RPMI 1640 medium (RPMI), and synthetic amino acid medium for fungi (SAAMF). Inocula were standardized spectrophotometrically, and endpoints were defined by the complete absence of growth for AMB and by no more than 25% of the growth in the drug-free control for all other agents. Comparative analyses of median MICs, as determined by each test method, were made for all drug-medium combinations. Both methods yielded similar (+/- 1 twofold dilution) median MICs for AMB in EMEM and RPMI, 5FC in all media, and FLU in EMEM, RPMI, and SAAMF. In contrast, substantial between-method variations in median MICs were seen for AMB in BYNB and SAAMF, FLU In BYNB, and ITRA and SCH in all media. Interlaboratory concordance of median MICs was good for AMB, 5FC, and FLU but poor for ITRA and SCH in all media. Endpoint determinations were analyzed by use of kappa statistical analyses for evaluating the strength of observer agreement. Moderate to almost perfect interlaboratory agreement occurred with AMB and 5FC in all media and with FLU in EMEM, RPMI, and SAAMF, irrespective of the test method. Slight to almost perfect interlaboratory agreement occurred with ITRA and SCH in EMEM, RPMI, and SAAMF when tested by semisolid agar dilution but not broth microdilution. Kappa values assessing intralaboratory agreement between methods were high for 5FC in all media, for AMB in BYNB, ENEM, and RPMI, and for FLU in EMEM, RPMI, and SAAMF. One laboratory

  12. Determination of in vitro synergy for dual antimicrobial therapy against resistant Neisseria gonorrhoeae using Etest and agar dilution.

    PubMed

    Wind, Carolien M; de Vries, Henry J C; van Dam, Alje P

    2015-03-01

    In response to antimicrobial resistance of Neisseria gonorrhoeae to last-resort extended-spectrum cephalosporins, combination therapy of azithromycin+ceftriaxone is now recommended. Dual therapy can be effective to treat monoresistant strains as well as multidrug-resistant strains, preferably employing the effect of in vitro synergy. As reports on in vitro synergy of azithromycin+ceftriaxone in N. gonorrhoeae are conflicting, in this study an evaluation of this combination was performed using a cross-wise Etest method and agar dilution. Synergy was defined as a fractional inhibitory concentration index (FICI) of ≤0.5. To identify other dual treatment options for gonorrhoea, in vitro synergy was evaluated for 65 dual antimicrobial combinations using Etest. Azithromycin, cefixime, ceftriaxone, colistin, ertapenem, fosfomycin, gentamicin, minocycline, moxifloxacin, rifampicin, spectinomycin and tigecycline were screened for synergy in all possible combinations. No synergy or antagonism was found for any of the 65 combinations. The geometric mean FICI ranged from 0.82 to 2.00. The mean FICI of azithromycin+ceftriaxone was 1.18 (Etest) and 0.55 (agar dilution). The difference between both methods did not result in a difference in interpretation of synergy. Ceftriaxone-resistant strain F89 was tested in all combinations and no synergy was found for any of them. Most importantly, the ceftriaxone minimum inhibitory concentration of F89 was not decreased below the breakpoint with any concentration of azithromycin.

  13. Estimation method for serial dilution experiments.

    PubMed

    Ben-David, Avishai; Davidson, Charles E

    2014-12-01

    Titration of microorganisms in infectious or environmental samples is a corner stone of quantitative microbiology. A simple method is presented to estimate the microbial counts obtained with the serial dilution technique for microorganisms that can grow on bacteriological media and develop into a colony. The number (concentration) of viable microbial organisms is estimated from a single dilution plate (assay) without a need for replicate plates. Our method selects the best agar plate with which to estimate the microbial counts, and takes into account the colony size and plate area that both contribute to the likelihood of miscounting the number of colonies on a plate. The estimate of the optimal count given by our method can be used to narrow the search for the best (optimal) dilution plate and saves time. The required inputs are the plate size, the microbial colony size, and the serial dilution factors. The proposed approach shows relative accuracy well within ±0.1log10 from data produced by computer simulations. The method maintains this accuracy even in the presence of dilution errors of up to 10% (for both the aliquot and diluent volumes), microbial counts between 10(4) and 10(12) colony-forming units, dilution ratios from 2 to 100, and plate size to colony size ratios between 6.25 to 200.

  14. Comparison of agar dilution and antibiotic gradient strip test with broth microdilution for susceptibility testing of swine Brachyspira species.

    PubMed

    Mirajkar, Nandita S; Gebhart, Connie J

    2016-03-01

    Production-limiting diseases in swine caused by Brachyspira are characterized by mucohemorrhagic diarrhea (B. hyodysenteriae and "B. hampsonii") or mild colitis (B. pilosicoli), while B. murdochii is often isolated from healthy pigs. Emergence of novel pathogenic Brachyspira species and strains with reduced susceptibility to commonly used antimicrobials has reinforced the need for standardized susceptibility testing. Two methods are currently used for Brachyspira susceptibility testing: agar dilution (AD) and broth microdilution (BMD). However, these tests have primarily been used for B. hyodysenteriae and rarely for B. pilosicoli. Information on the use of commercial susceptibility testing products such as antibiotic gradient strips is lacking. Our main objective was to validate and compare the susceptibility results, measured as the minimum inhibitory concentration (MIC), of 6 antimicrobials for 4 Brachyspira species (B. hyodysenteriae, "B. hampsonii", B. pilosicoli, and B. murdochii) by BMD and AD (tiamulin, valnemulin, lincomycin, tylosin, and carbadox) or antibiotic gradient strip (doxycycline) methods. In general, the results of a high percentage of all 4 Brachyspira species differed by ±1 log2 dilution or less by BMD and AD for tiamulin, valnemulin, lincomycin, and tylosin, and by BMD and antibiotic gradient strip for doxycycline. The carbadox MICs obtained by BMD were 1-5 doubling dilutions different than those obtained by AD. BMD for Brachyspira was quicker to perform with less ambiguous interpretation of results when compared with AD and antibiotic gradient strip methods, and the results confirm the utility of BMD in routine diagnostics. PMID:26965233

  15. Comparison of Neisseria gonorrhoeae MICs obtained by Etest and agar dilution for ceftriaxone, cefpodoxime, cefixime and azithromycin.

    PubMed

    Gose, Severin; Kong, Carol J; Lee, Yer; Samuel, Michael C; Bauer, Heidi M; Dixon, Paula; Soge, Olusegun O; Lei, John; Pandori, Mark

    2013-12-01

    We evaluated Neisseria gonorrhoeae Etest minimum inhibitory concentrations (MICs) relative to agar dilution MICs for 664 urethral isolates for ceftriaxone (CRO) and azithromycin (AZM), 351 isolates for cefpodoxime (CPD) and 315 isolates for cefixime (CFM). Etest accurately determined CPD, CFM and AZM MICs, but resulted in higher CRO MICs.

  16. Comparison of Neisseria gonorrhoeae MICs Obtained by Etest and Agar Dilution for Ceftriaxone, Cefpodoxime, Cefixime and Azithromycin.

    PubMed

    Gose, Severin; Kong, Carol J; Lee, Yer; Samuel, Michael C; Bauer, Heidi M; Dixon, Paula; Soge, Olusegun O; Lei, John; Pandori, Mark

    2013-10-24

    We evaluated Neisseria gonorrhoeae Etest minimum inhibitory concentrations (MICs) relative to agar dilution MICs for 664 urethral isolates for ceftriaxone (CRO) and azithromycin (AZM), 351 isolates for cefpodoxime (CPD) and 315 isolates for cefixime (CFM). Etest accurately determined CPD, CFM and AZM MICs, but resulted in higher CRO MICs.

  17. Antimicrobial susceptibility testing for Helicobacter pylori isolates from Brazilian children and adolescents: Comparing agar dilution, E-test, and disk diffusion

    PubMed Central

    Ogata, Silvio Kazuo; Gales, Ana Cristina; Kawakami, Elisabete

    2014-01-01

    Antimicrobial susceptibility testing for Helicobacter pylori is increasingly important due to resistance to the most used antimicrobials agents. Only agar dilution method is approved by CLSI, but it is difficult to perform routinely. We evaluated the reliability of E-test and disk diffusion comparing to agar dilution method on Helicobacter pylori antimicrobial susceptibility testing. Susceptibility testing was performed for amoxicillin, clarithromycin, furazolidone, metronidazole and tetracycline using E-test, disk-diffusion and agar dilution method in 77 consecutive Helicobacter pylori strains from dyspeptic children and adolescents. Resistance rates were: amoxicillin - 10.4%, 9% and 68.8%; clarithromycin - 19.5%, 20.8%, 36.3%; metronidazole - 40.2%33.7%, 38.9%, respectively by agar dilution, E-test and disk diffusion method. Furazolidone and tetracycline showed no resistance rates. Metronidazole presented strong correlation to E-test (r = 0.7992, p < 0.0001) and disk diffusion method (r=-0.6962, p < 0.0001). Clarithromycin presented moderate correlation to E-test (r = 0.6369, p < 0.0001) and disk diffusion method (r=−0.5656, p < 0.0001). Amoxicillin presented weak correlation to E-test (r = 0.3565, p = 0.0015) and disk diffusion (r=−0.3565, p = 0.0015). Tetracycline presented weak correlation with E-test (r = 0.2346, p = 0.04) and furazolidone to disk diffusion (r=−0.0288, p = 0.8038). E-test presented better agreement with gold standard. It is an easy and reliable method for Helicobacter pylori susceptibility testing. Disk diffusion method presented high disagreement and high rates of major errors. PMID:25763052

  18. Method for Measuring Changes in Surface Tension on Agar

    PubMed Central

    Weisberg, David S.; Dworkin, Martin

    1983-01-01

    The surface tension of agar surfaces was determined by measuring the contact angles formed by drops of various hydrophobic liquids on the surface and then calculating the composite surface free energy function by solving a series of simultaneous equations derived from these data. This method was used to measure the change in the surface tension of agar produced by the addition of various concentrations of albumin. The resulting curve was typical of the effect of increasing concentrations of surfactants on surface tension. The method was compared with other methods of determining surface tension of solids, and it was concluded that the technique used here provided the most reliable results. PMID:16346273

  19. Dispersion serial dilution methods using the gradient diluter device.

    PubMed

    Walling, Leslie; Schulz, Craig; Johnson, Michael

    2012-12-01

    A solute aspirated into a prefilled tube of diluent undergoes a dilution effect known as dispersion. Traditionally the effects of dispersion have been considered a negative consequence of using liquid-filled fixed-tip liquid handlers. We present a novel device and technique that utilizes the effects of dispersion to the benefit of making dilutions. The device known as the Gradient Diluter extends the dilution range of practical serial dilutions to six orders of magnitude in final volumes as low as 10 μL. Presented are the device, dispersion methods, and validation tests using fluorescence detection of sulforhodamine and the high-performance liquid chromatography/ultraviolet detection of furosemide. In addition, a T-cell inhibition assay of a relevant downstream protein is used to demonstrate IC(50) curves made with the Gradient Diluter compare favorably with those generated by hand.

  20. Oxacillin susceptibility testing of Staphylococcus saprophyticus using disk diffusion, agar dilution, broth microdilution, and the Vitek GPS-105 card.

    PubMed

    Ramotar, K; Woods, W; Toye, B

    2001-08-01

    Eighty-three mecA negative isolates of S. saprophyticus had oxacillin zone diameters agar dilution, broth microdilution, or the Vitek GPS-105 card. Greater than 90% of these isolates would be considered resistant using NCCLS M7-A5, M100-S10 criteria. These results suggest that the current NCCLS MIC and zone diameter breakpoints for oxacillin resistance in coagulase-negative Staphylococci are not appropriate for S. saprophyticus as they do not correlate with the presence of the mecA gene.

  1. A modified agar plate method for detection of Strongyloides stercoralis.

    PubMed

    Koga, K; Kasuya, S; Khamboonruang, C; Sukhavat, K; Ieda, M; Takatsuka, N; Kita, K; Ohtomo, H

    1991-10-01

    The agar plate method is a new technique with high detection rates for coprological diagnosis of human strongyloidiasis. This report details modifications of the technique and establishes a standardized procedure. We recommend that all plates should be carefully observed using a microscope because macroscopic observation can lead to false negative results. It is also advisable to pour formalin solution directly into microscopically positive dishes to collect worms by sedimentation. This procedure enables one to observe worms otherwise hidden. Sealing dishes with adhesive tape prevents larvae from crawling out of the dishes, eliminating any possibility in the reduction of detection rates, and greatly improves the safety conditions for the technician performing the procedure. We consider the agar plate method to be superior to the filter paper method in detecting Strongyloides, and we believe that it will eventually become the technique of choice. PMID:1951861

  2. Standardization of disk diffusion and agar dilution susceptibility tests for Neisseria gonorrhoeae: interpretive criteria and quality control guidelines for ceftriaxone, penicillin, spectinomycin, and tetracycline.

    PubMed Central

    Jones, R N; Gavan, T L; Thornsberry, C; Fuchs, P C; Gerlach, E H; Knapp, J S; Murray, P; Washington, J A

    1989-01-01

    A six-laboratory study developed a standardized method for determining the susceptibilities of Neisseria gonorrhoeae strains to penicillin, tetracycline, spectinomycin, and ceftriaxone. Three quality control organisms were also selected, and quality assurance guidelines were initially generated for the disk diffusion and agar dilution methods. The medium recommended for gonococcal susceptibility testing was GC agar with a defined "XV-like" supplement. The supplement should be free of cysteine, a component implicated in the inactivation of some newer beta-lactam compounds. Penicillin, tetracycline, spectinomycin, and ceftriaxone were stable in agar plates stored at 3 to 5 degrees C for at least 2 weeks. Numerous GC agar and drug disk lots were used during the trials without significant variation in test results. Several other gonococcal strains were recommended for additional medium quality assurance. The disk quality control zone limits were established for N. gonorrhoeae ATCC 49226 (formerly CDC F-18) and Staphylococcus aureus ATCC 25923. MIC quality control ranges were also developed for N. gonorrhoeae ATCC 49226 and S. aureus ATCC 29213. The interpretive criteria for penicillin were as follows: susceptibility, greater than or equal to 47 mm (diameter of inhibition zone) (less than or equal to 0.06 micrograms/ml [MIC]); resistance, less than or equal to 26 mm (greater than or equal to 2 micrograms/ml). For tetracycline they were as follows: susceptibility, greater than or equal to 38 mm (less than or equal to 0.25 microgram/ml); resistance, less than or equal to 30 mm (greater than or equal to 2 micrograms/ml). For spectinomycin they were as follows: susceptibility, >/= 18 mm (/= 128 micrograms/ml). For ceftriaxone susceptibility, the criterion was >/= 35 mm (

  3. Antimicrobial susceptibility testing of Australian isolates of Brachyspira hyodysenteriae using a new broth dilution method.

    PubMed

    Karlsson, Märit; Oxberry, Sophy L; Hampson, David J

    2002-01-01

    The antimicrobial susceptibilities of 76 field isolates of Brachyspira hyodysenteriae from different states of Australia were tested in a newly developed broth dilution procedure. The antimicrobial agents used were tiamulin, valnemulin, tylosin, erythromycin, lincomycin and clindamycin. The results from the broth dilution susceptibility testing of 39 of the isolates were compared with results obtained for the same isolates using the agar dilution method. Amongst the isolates tested by broth dilution, 17 were from three farms and had been collected over a number of years. Their pulsed field gel electrophoresis pattern previously had been determined. The broth dilution technique was simple to use, less labor intensive than agar dilution, and gave clear end points. The results obtained using the two methods generally corresponded well, although in a few cases the MIC obtained by broth dilution were lower than those with agar dilution. For the 76 isolates tested by broth dilution, the MIC(90) (mg/l) was: tiamulin, 1; valnemulin, 0.5; tylosin>256; erythromycin>256; lincomycin, 64 and clindamycin, 16. Only minor differences in susceptibility patterns were found amongst isolates from different Australian states. Over all the isolates, and also amongst the isolates obtained from different years on the three farms, there was no trend for the susceptibility of the isolates to alter with time. PMID:11731165

  4. Total Antioxidant Capacity of Serum Determined Using the Potassium Permanganate Agar Method Based on Serum Diffusion in Agar

    PubMed Central

    Zhou, Ying; Zhang, Meijuan; Liu, Hui

    2015-01-01

    Objectives. To develop a new method for determining total antioxidants in serum and to evaluate the total antioxidant capacity of organisms. Design and Methods. Sodium hyposulfite (Na2S2O3) and serum were used to evaluate the linearity and precision of the potassium permanganate agar method. The area of serum diffusion in samples from 30 intensive care unit (ICU) patients compared with 44 healthy subjects was determined by the potassium permanganate agar method. Results. The linearity (R2 in the linear experiment of Na2S2O3 was 0.994; R2 in the linear experiment of serum was 0.987) and precision (coefficient of variation of area of high level serum diffusion within-run, between-run, and between-day and coefficient of variation of area of low serum diffusion within-run, between-run, and between-day were all less than 10%) were acceptable using the potassium permanganate agar method. Total antioxidants of serum between the ICU group and the healthy group were different (p = 0.002, two tailed). Conclusions. Total antioxidants in serum can be determined by the potassium permanganate agar method. The total antioxidant capacity of an organism can be evaluated by the amount of total antioxidants in serum. PMID:26347595

  5. The Resazurin-Agar Method - a Quick Test to Determine Water Quality

    NASA Astrophysics Data System (ADS)

    Huckfeldt, J.; Westphal, B.; Claußen, L.

    2015-12-01

    Rezasurin has been used as a smart tracer in stream ecosystems to indicate metabolic activity, specifically aerobic respiration by heterotrophic bacteria. Resazurin is a blue compound which is irreversibly reduced to the pink resorufin in the presence of aerobic bacteria. The degree and speed of colour change from blue to pink is a measure of the degree of oxygen consumption and thus an indirect indication of the concentration of aerobic bacteria in a given medium. A high concentration of bacteria in water indicates a bad water quality. In our work a method was developed using resazurin agar plates to find a quick and easy way for testing water quality and comparing concentrations of bacteria in freshwater and seawater samples. The theory was to concentrate bacteria from a defined volume of water sample onto polycarbonate filters (0.2 μm), which are then placed onto the resazurin agar plate. The presence of aerobic bacteria on the filter will reduce the resazurin in the agar and the compound changes its colour. First tests conducted with different dilutions of a pure culture of yoghurt bacteria showed promising results and confirmed the feasibility of the method. In a further assay, we used water samples from different water layers and different temperatures and were also able to observe differences in the concentration of bacteria, depending on these different environmental conditions.The assay was also successfully used with seawater samples, collected from 2 different stations at 3 different depths in the Baltic Sea (salinity=15). The discolouration of the plates showed good correlation with the oxygen concentrations in the water. The resazurin-agar plate method is economical and fast. Several samples could be investigated at the same time without sacrificing the reliability of the results. Thus it is a good pre-screening test for a quantitative evaluation of bacteria in a water sample.

  6. Agar underlay method for recovery of sublethally heat-injured bacteria.

    PubMed

    Kang, D H; Siragusa, G R

    1999-12-01

    A method of recovering sublethally heat-injured bacteria was developed. The procedure (termed the agar underlay method) uses a nonselective agar underlaid with a selective medium. In a two-chambered petri dish, the Lutri plate (LP), a nonselective agar is inoculated with a population of sublethally heat-injured bacteria. After a 2-h repair incubation period, selective agar is added to the bottom chamber of the LP and incubated. By diffusing through the nonselective top agar, selective agents from the underlay medium impart selectivity to the system. By the agar underlay method, recovery rates of the heat-injured food-borne pathogens Escherichia coli O157:H7 and Salmonella typhimurium were not different (P > 0. 05) from recovery rates determined with nonselective media. Sublethally heat-injured cells (60 degrees C for 1.5 min in buffer or 80 degrees C for 30 s on meat surfaces) grew and produced a typical colony morphology and color reaction when the agar underlay procedure was used with the appropriate respective selective agars. Unlike agar overlay methods for injury repair, the agar underlay procedure allows the typical selective-medium colony morphology to develop and allows colonies to be more easily picked for further characterization. Higher recovery rates of heat-injured fecal enterococci from bovine fecal samples and total coliforms from animal waste lagoons were obtained by the agar underlay method with selective agars than by direct plating on the respective selective media. PMID:10583985

  7. An extension of the Coconut Cream Agar method to screen Penicillium citrinum isolates for citrinin production.

    PubMed

    Mohamed, S; Flint, S; Palmer, J; Fletcher, G C; Pitt, J I

    2013-09-01

    A simple and rapid screening method was developed for the detection of citrinin in fungal cultures using Coconut Cream Agar (CCA) described previously for detecting aflatoxin and ochratoxin A. Fifteen isolates of Penicillium citrinum were inoculated onto CCA and incubated at 25 and 30°C for 10 days. All isolates produced a distinct yellow green fluorescence on CCA when the reverse side of the agar plates were viewed under long wavelength UV light. Detection was optimal at 25°C after four to 5 days of incubation. Isolates positive by the CCA method also tested positive for citrinin production by the TLC agar plug method after growth on CCA, Czapek yeast extract agar and yeast extract sucrose agar. Control cultures were negative by both methods, indicating that the CCA Petri dish method was suitable for screening cultures for citrinin production.

  8. A solid agar overlay method for recovery of heat-injured Listeria monocytogenes.

    PubMed

    Yan, Zhinong; Gurtler, Joshua B; Kornacki, Jeffrey L

    2006-02-01

    A solid agar overlay method was developed for recovery of heat-injured Listeria monocytogenes. Presolidified nonselective tryptic soy agar with 0.6% yeast extract (TSAYE, 2% agar) was overlaid on top of solidified modified Oxford agar (MOX). Heat injury of L. monocytogenes was conducted at 58 degrees C for 6 min in a jacketed flask filled with tryptic soy broth. Both noninjured and heat-treated L. monocytogenes cells were plated onto TSAYE, MOX, and TSAYE-MOX plates. No significant differences (P > 0.05) in recovery were found among the three media for noninjured bacterial cells. Recovery of heat-injured L. monocytogenes cells on TSAYE-MOX overlay plates was equivalent to that on the nonselective TSAYE medium, whereas recovery on the selective MOX medium was significantly lower (P < 0.05) compared with both TSAYE and the overlay plates. There were no significant differences (P > 0.05) among the overlay plates prepared 0, 2, 4, 6, 8, 16, and 24 h prior to plating heat-injured bacterial cells. The TSAYE-MOX overlay also allowed differentiation of L. monocytogenes from a mixture of four other types of foodborne pathogens. This solid agar overlay method for recovery of heat-injured L. monocytogenes cells is less time-consuming and less complicated than the conventional overlay-underlay technique and the double overlay modification of the thin agar layer method and may allow for greater laboratory plating efficiencies.

  9. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2004-01-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Anionic surfactants (SS-6656, Alfoterra 35, 38, 63,65,68) have been identified which can change the wettability of the calcite surface to intermediate/water-wet condition as well or better than the cationic surfactant DTAB with a West Texas crude oil in the presence of Na{sub 2}CO{sub 3}. All the carbonate surfaces (Lithographic Limestone, Marble, Dolomite and Calcite) show similar behavior with respect to wettability alteration with surfactant 4-22. Anionic surfactants (5-166, Alfoterra-33 and Alfoterra-38 and Alfoterra-68), which lower the interfacial tension with a West Texas crude oil to very low values (<10{sup -2} nM/m), have also been identified. Plans for the next quarter include conducting wettability, mobilization, and imbibition studies.

  10. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2003-07-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. We have conducted adsorption, phase behavior and wettability studies. Addition of Na{sub 2}CO{sub 3} decreases IFT with a minimum at about 0.2 M. Addition of surfactant decreases IFT further. In the absence of surfactant the minerals are oil wet after aging with crude oil. Addition of surfactant solution decreases the contact angle to intermediate wettability. Addition of Na{sub 2}CO{sub 3} decreases anionic surfactant adsorption on calcite surface. Plans for the next quarter include conducting adsorption, phase behavior and wettability studies.

  11. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2005-07-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Laboratory imbibition tests show that imbibition rate is not very sensitive to the surfactant concentration (in the range of 0.05-0.2 wt%) and small amounts of trapped gas saturation. It is however very sensitive to oil permeability and water-oil-ratio. Less than 0.5 M Na2CO3 is needed for in situ soap generation and low adsorption; NaCl can be added to reach the necessary total salinity. The simulation result matches the laboratory imbibition experimental data. Small fracture spacing and high permeability would be needed for high rate of recovery.

  12. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2003-10-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. We have conducted adsorption, phase behavior, interfacial tension (IFT) and wettability studies. Alfoterra-38 (0.05 wt%), Alfoterra-35 (0.05 wt%), SS-6656 (0.05 wt%), and DTAB (1 wt%) altered the wettability of the initially oil-wet calcite plate to an intermediate/water-wet state. Low IFT ({approx}10{sup -3} dynes/cm) is obtained with surfactants 5-166, Alfoterra-33 and Alfoterra-38. Plans for the next quarter include conducting wettability and mobilization studies.

  13. Dilute Surfactant Methods for Carbonate Formations

    SciTech Connect

    Kishore K. Mohanty

    2004-03-31

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Anionic surfactants (Alfoterra 35, 38) recover more than 40% of the oil in about 50 days by imbibition driven by wettability alteration in the core-scale. Anionic surfactant, Alfoterra-68, recovers about 28% of the oil by lower tension aided gravity-driven imbibition in the core-scale. Residual oil saturation showed little capillary number dependence between 10{sup -5} and 10{sup -2}. Wettability alteration increases as the number of ethoxy groups increases in ethoxy sulfate surfactants. Plans for the next quarter include conducting mobilization, and imbibition studies.

  14. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2004-10-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Simulation studies indicate that both wettability alteration and gravity-driven flow play significant role in oil recovery from fractured carbonates. Anionic surfactants (Alfoterra 35, 38) recover about 55% of the oil in about 150 days by imbibition driven by wettability alteration and low tension in the core-scale. Anionic surfactant, Alfoterra-68, recovers about 40% of the oil by lower tension aided gravity-driven imbibition in the core-scale. Cationic surfactant, DTAB recovers about 35% of the oil. Plans for the next quarter include conducting simulation and imbibition studies.

  15. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2005-01-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Imbibition in an originally oil-wet 2D capillary is the fastest in the case of Alf-38 and slowest in the case of DTAB (among the surfactants studied). Force of adhesion studies and contact angle measurements show that greater wettability alteration is possible with these anionic surfactants than the cationic surfactant studied. The water imbibition rate does not increase monotonically with an increase in the surfactant concentration. A numerical model has been developed that fits the rate of imbibition. Plans for the next quarter include conducting simulation and imbibition studies.

  16. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2003-01-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. We have acquired field oil and core samples and field brine compositions from Marathon. We have conducted preliminary adsorption and wettability studies. Addition of Na{sub 2}CO{sub 3} decreases anionic surfactant adsorption on calcite surface. Receding contact angles increase with surfactant adsorption. Plans for the next quarter include conducting adsorption, phase behavior and wettability studies.

  17. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2003-07-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. We have conducted adsorption, phase behavior, interfacial tension (IFT) and wettability studies. Addition of Na{sub 2}CO{sub 3} decreases IFT with a minimum at about 0.2 M. Addition of surfactant decreases IFT further. In the absence of surfactant the minerals are oil-wet after aging with crude oil. Addition of surfactant solution decreases the contact angle to intermediate-wet for many surfactants and water-wet for one surfactant. Addition of Na{sub 2}CO{sub 3} decreases anionic surfactant adsorption on calcite surface. Plans for the next quarter include conducting core adsorption, phase behavior, wettability and mobilization studies.

  18. Dilute Surfactant Methods for Carbonate Formations

    SciTech Connect

    Kishore K. Mohanty

    2005-10-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the best hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Laboratory-scale surfactant brine imbibition experiments give high oil recovery (35-62% OOIP) for initially oil-wet cores through wettability alteration and IFT reduction. Core-scale simulation results match those of the experiments. Initial capillarity-driven imbibition gives way to a final gravity-driven process. As the matrix block height increases, surfactant alters wettability to a lesser degree, or permeability decreases, oil production rate decreases. The scale-up to field scale will be further studied in the next quarter.

  19. DILUTE SURFACTANT METHODS FOR CARBONATE FORMATIONS

    SciTech Connect

    Kishore K. Mohanty

    2005-04-01

    There are many carbonate reservoirs in US (and the world) with light oil and fracture pressure below its minimum miscibility pressure (or reservoir may be naturally fractured). Many carbonate reservoirs are naturally fractured. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). Thermal and miscible tertiary recovery techniques are not effective in these reservoirs. Surfactant flooding (or huff-n-puff) is the only hope, yet it was developed for sandstone reservoirs in the past. The goal of this research is to evaluate dilute (hence relatively inexpensive) surfactant methods for carbonate formations and identify conditions under which they can be effective. Laboratory imbibition tests show about 61% oil recovery in the case of Alf-38 and 37% in the case of DTAB. A numerical model has been developed that fits the rate of imbibition of the laboratory experiment. Field-scale fracture block simulation shows that as the fracture spacing increases, so does the time of recovery. Plans for the next quarter include simulation studies.

  20. Dilute Surfactant Methods for Carbonate Formations

    SciTech Connect

    Kishore K. Mohanty

    2006-02-01

    There are many fractured carbonate reservoirs in US (and the world) with light oil. Waterflooding is effective in fractured reservoirs, if the formation is water-wet. Many fractured carbonate reservoirs, however, are mixed-wet and recoveries with conventional methods are low (less than 10%). The process of using dilute anionic surfactants in alkaline solutions has been investigated in this work for oil recovery from fractured oil-wet carbonate reservoirs both experimentally and numerically. This process is a surfactant-aided gravity drainage where surfactant diffuses into the matrix, lowers IFT and contact angle, which decrease capillary pressure and increase oil relative permeability enabling gravity to drain the oil up. Anionic surfactants have been identified which at dilute concentration of 0.05 wt% and optimal salinity can lower the interfacial tension and change the wettability of the calcite surface to intermediate/water-wet condition as well or better than the cationic surfactant DTAB with a West Texas crude oil. The force of adhesion in AFM of oil-wet regions changes after anionic surfactant treatment to values similar to those of water-wet regions. The AFM topography images showed that the oil-wetting material was removed from the surface by the anionic surfactant treatment. Adsorption studies indicate that the extent of adsorption for anionic surfactants on calcite minerals decreases with increase in pH and with decrease in salinity. Surfactant adsorption can be minimized in the presence of Na{sub 2}CO{sub 3}. Laboratory-scale surfactant brine imbibition experiments give high oil recovery (20-42% OOIP in 50 days; up to 60% in 200 days) for initially oil-wet cores through wettability alteration and IFT reduction. Small (<10%) initial gas saturation does not affect significantly the rate of oil recovery in the imbibition process, but larger gas saturation decreases the oil recovery rate. As the core permeability decreases, the rate of oil recovery reduces

  1. Efficacy of the thin agar layer method for the recovery of stressed Cronobacter spp. (Enterobacter sakazakii).

    PubMed

    Osaili, Tareq M; Al-Nabulsi, Anas A; Shaker, Reyad R; Al-Holy, Murad M; Al-Haddaq, Mohammed S; Olaimat, Amin N; Ayyash, Mutamed M; Al Ta'ani, Mahmoud K; Forsythe, Stephen J

    2010-10-01

    Cronobacter spp. (Enterobacter sakazakii) are emerging opportunistic pathogens for all age groups, and are of particular concern when it comes to infants. Prior to contaminating food, the organism may be exposed to a variety of stresses, leading to a generation of sublethally injured cells that may not be detected by selective media unless a protracted recovery period is included in the isolation procedure. This study evaluated the efficacy of the thin agar layer (TAL) method for the recovery of Cronobacter cells that had been exposed to various stress conditions. Five strains of C. sakazakii and C. muytjensii were exposed to starvation, heat, cold, acid, alkaline, chlorine, or ethanol, with or without further exposure to desiccation stress. The recovery of the stressed cells was determined on tryptone soy agar (TSA; nonselective control medium), violet red bile glucose agar (VRBGA; selective agar), Druggan-Forsythe-Iversen (DFI; selective agar), and TAL media (viz., VRBGA overlaid with TSA, and DFI overlaid with TSA). Regardless of stress type, there were no significant differences among the recoveries of stressed desiccated Cronobacter spp. cultures on TSA, DFI+TSA, and VRBGA+TSA, but there was significantly less recovery on VRBGA. The recovery of prestressed desiccated Cronobacter spp. on DFI+TSA was similar to that on TSA, whereas the recovery on VRBGA+TSA was lower. DFI+TSA performed better than VRBGA+TSA did in differentiating Cronobacter spp. within mixed bacterial cultures. The results of this study suggest the use of the TAL method DFI+TSA as an improved method for the direct recovery of stressed Cronobacter spp.

  2. Correlation of agar dilution and VITEK2 system for detection of resistance to macrolides, lincosamides and pristinamycin among Staphylococcus aureus and Staphylococcus epidermidis: association with genotypes.

    PubMed

    Bémer, P; Juvin, M-E; Corvec, S; Ros, A; Drugeon, H

    2005-08-01

    The performance of the VITEK2 system was evaluated against the agar dilution reference procedure for testing susceptibility of Staphylococcus aureus and Staphylococcus epidermidis to macrolides, lincosamides and streptogramins (MLS). Eighty clinical isolates were selected according to their resistance phenotype and genotype. Results for erythromycin and clindamycin showed 100% agreement; results for lincomycin showed agreement of 78%, with one very major error and 17 minor errors; and results for pristinamycin showed agreement of 46%, with one major error and 43 minor errors. Most isolates resistant to lincomycin and streptogramin A (L SgAr phenotype) were falsely susceptible to lincomycin, and intermediately-resistant or resistant to pristinamycin, with the VITEK2 system. No resistance gene was detected. Most (80%) isolates resistant constitutively to MLS (MLS(r)BC phenotype) were falsely intermediately-resistant to pristinamycin with the VITEK2 system. The erm(A) gene was more common than erm(C) in MLS(r)BC strains. Resistance to pristinamycin alone (SgA SgB PTr phenotype), or associated with either lincomycin resistance (L SgA SgB PTr phenotype) or constitutive MLS(B) resistance (MLS(BC) SgA PTr phenotype), was well-characterised without discordant results. Resistance to pristinamycin was always associated with resistance to streptogramin A, encoded by the vga(A), vga(B), vgb(A) and vat(A) genes in association with the erm(A) or erm(C) genes.

  3. Correlation of agar dilution and VITEK2 system for detection of resistance to macrolides, lincosamides and pristinamycin among Staphylococcus aureus and Staphylococcus epidermidis: association with genotypes.

    PubMed

    Bémer, P; Juvin, M-E; Corvec, S; Ros, A; Drugeon, H

    2005-08-01

    The performance of the VITEK2 system was evaluated against the agar dilution reference procedure for testing susceptibility of Staphylococcus aureus and Staphylococcus epidermidis to macrolides, lincosamides and streptogramins (MLS). Eighty clinical isolates were selected according to their resistance phenotype and genotype. Results for erythromycin and clindamycin showed 100% agreement; results for lincomycin showed agreement of 78%, with one very major error and 17 minor errors; and results for pristinamycin showed agreement of 46%, with one major error and 43 minor errors. Most isolates resistant to lincomycin and streptogramin A (L SgAr phenotype) were falsely susceptible to lincomycin, and intermediately-resistant or resistant to pristinamycin, with the VITEK2 system. No resistance gene was detected. Most (80%) isolates resistant constitutively to MLS (MLS(r)BC phenotype) were falsely intermediately-resistant to pristinamycin with the VITEK2 system. The erm(A) gene was more common than erm(C) in MLS(r)BC strains. Resistance to pristinamycin alone (SgA SgB PTr phenotype), or associated with either lincomycin resistance (L SgA SgB PTr phenotype) or constitutive MLS(B) resistance (MLS(BC) SgA PTr phenotype), was well-characterised without discordant results. Resistance to pristinamycin was always associated with resistance to streptogramin A, encoded by the vga(A), vga(B), vgb(A) and vat(A) genes in association with the erm(A) or erm(C) genes. PMID:16008619

  4. Colony morphotype on Sabouraud-triphenyltetrazolium agar: a simple and inexpensive method for Candida subspecies discrimination.

    PubMed Central

    Quindós, G; Fernández-Rodríguez, M; Burgos, A; Tellaetxe, M; Cisterna, R; Pontón, J

    1992-01-01

    A new method of Candida subspecies discrimination on Sabouraud-triphenyltetrazolium agar is reported. Five hundred sixty-two strains of Candida and Torulopsis glabrata, previously identified by conventional mycological methods, were studied. Each strain received a three-letter code and a number based on its colonial morphology. Sixteen morphotypes were found for Candida albicans, 6 were found for Candida parapsilosis, 4 were found for both Candida guilliermondii and Candida krusei, and 12 were found for Candida tropicalis. None of the 56 T. glabrata strains studied grew on this agar. A reproducibility of 95% was found for C. albicans. The simplicity and low cost could make this method useful for typing Candida spp. Images PMID:1400981

  5. Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis.

    PubMed

    Kaiser, Thaís Dias Lemos; Pereira, Eliezer Menezes; Dos Santos, Kátia Regina Netto; Maciel, Ethel Leonor Noia; Schuenck, Ricardo Pinto; Nunes, Ana Paula Ferreira

    2013-03-01

    Staphylococcus epidermidis in immunocompromised patients can cause bacteremia related to the use of catheter due to biofilm production. There are different phenotypic methods to detect biofilm formation. One method is based on culture in brain heart infusion agar (BHIA) containing sucrose and red Congo dye (original Congo red agar). Our group created a new CRA formula and we have confirmed its capacity to detect biofilm production in 210 S. epidermidis strains, including 76 (36.2%) icaAB gene-positive strains. Other parameters were also evaluated. The new CRA formula that gave the best results was BHIA with sucrose (5%), Congo red (0.08%), NaCl (1.5%), glucose (2%), and vancomycin (0.5 mg/mL) (vancomycin-modified CRA-CRAmod). The CRAmod plus vancomycin may be a promising tool and can help to determine the real participation of S. epidermidis in the infectious process.

  6. Modification of the Congo red agar method to detect biofilm production by Staphylococcus epidermidis.

    PubMed

    Kaiser, Thaís Dias Lemos; Pereira, Eliezer Menezes; Dos Santos, Kátia Regina Netto; Maciel, Ethel Leonor Noia; Schuenck, Ricardo Pinto; Nunes, Ana Paula Ferreira

    2013-03-01

    Staphylococcus epidermidis in immunocompromised patients can cause bacteremia related to the use of catheter due to biofilm production. There are different phenotypic methods to detect biofilm formation. One method is based on culture in brain heart infusion agar (BHIA) containing sucrose and red Congo dye (original Congo red agar). Our group created a new CRA formula and we have confirmed its capacity to detect biofilm production in 210 S. epidermidis strains, including 76 (36.2%) icaAB gene-positive strains. Other parameters were also evaluated. The new CRA formula that gave the best results was BHIA with sucrose (5%), Congo red (0.08%), NaCl (1.5%), glucose (2%), and vancomycin (0.5 mg/mL) (vancomycin-modified CRA-CRAmod). The CRAmod plus vancomycin may be a promising tool and can help to determine the real participation of S. epidermidis in the infectious process. PMID:23313084

  7. A modified method for the detection of microbial proteases on agar plates using tannic acid.

    PubMed

    Saran, Saurabh; Isar, Jasmine; Saxena, Rajendra Kumar

    2007-06-10

    In routine assay for the screening of microbes producing proteases, 10% trichloroaceticacid (TCA) is flooded on the milk agar plates after inoculation and required incubation to precipitate the protein. However, the clarity of the hydrolyzed zone is not very sharp and distinct. We herein present an improved assay for detecting the presence of extracellular protease from microorganisms on agar plates. In this method 10% tannic acid is flooded on the milk agar plate (in place of, TCA) to observe the zone of hydrolysis. Tannic acid sharply increases the colour intensity of the plate, as it favours the precipitation of the unhydrolyzed protein in the plate, thereby improving the contrast between the intact zones and the enzymatic lyses zones of the substrate. Our results indicate that this method is useful to detect extracellular proteases produced by both fungi as well as bacteria. The method used in the present study is sensitive, and can be easily performed for screening of large number of microbial cultures. This is the first report on the use of tannic acid for the detection of microbial proteases.

  8. A method of test substance removal in agar colony assays using glass capillaries.

    PubMed

    Kastner, M; Maurer, H R

    1984-07-01

    A technique has been developed to remove test substances, after defined incubation periods, from clonogenic in vitro assays using agar-containing glass capillaries. Following removal from the capillaries, the entire agar gels were washed in petri dishes and redrawn into new capillaries. Using 8 radioactive biochemicals of molecular masses ranging from 150 to 1300 dalton the kinetics of diffusion between 1 and 20 min were determined. Using a wash solution-to-assay volume ratio of 20:1, a single washing for 10 min yielded between 90% and 99% removal by diffusion of test substances. By incorporating myelopoietic stem cells it was demonstrated that the cells to be assayed can be quantitatively transferred, without loss or stress, out of and back into capillaries. Thus the reversibility of test substance action can examined under defined conditions avoiding technical problems of previous methods.

  9. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus.

    PubMed

    Cold, Emma R; Freyria, Nastasia J; Martínez Martínez, Joaquín; Fernández Robledo, José A

    2016-01-01

    The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham's F12-5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham's F12-5% FBS- 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham's F12-5% FBS- 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications.

  10. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus

    PubMed Central

    Cold, Emma R.; Freyria, Nastasia J.; Martínez Martínez, Joaquín; Fernández Robledo, José A.

    2016-01-01

    The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham’s F12–5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham’s F12–5% FBS– 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham’s F12–5% FBS– 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. PMID:27149378

  11. An Agar-Based Method for Plating Marine Protozoan Parasites of the Genus Perkinsus.

    PubMed

    Cold, Emma R; Freyria, Nastasia J; Martínez Martínez, Joaquín; Fernández Robledo, José A

    2016-01-01

    The genus Perkinsus includes protozoan parasites of mollusks responsible for losses in the aquaculture industry and hampering the recovery of natural shellfish beds worldwide, and they are a key taxon for understanding intracellular parasitism adaptations. The ability to propagate the parasite in liquid media, in the absence of the host, has been crucial for improving understanding of its biology; however, alternative techniques to grow the parasite are needed to explore other basic aspects of the Perkinsus spp. biology. We optimized a DME: Ham's F12-5% FBS- containing solid agar medium for plating Perkinsus marinus. This solid medium supported trophozoite propagation both by binary fission and schizogony. Colonies were visible to the naked eye 17 days after plating. We tested the suitability of this method for several applications, including the following: 1) Subcloning P. marinus isolates: single discrete P. marinus colonies were obtained from DME: Ham's F12-5% FBS- 0.75% agar plates, which could be further propagated in liquid medium; 2) Subcloning engineered Perkinsus mediterraneus MOE[MOE]: GFP by streaking cultures on plates; 3) Chemical susceptibility: Infusing the DME: Ham's F12-5% FBS- 0.75% agar plates with triclosan resulted in inhibition of the parasite propagation in a dose-dependent manner. Altogether, our plating method has the potential for becoming a key tool for investigating diverse aspects of Perkinsus spp. biology, developing new molecular tools, and for biotechnological applications. PMID:27149378

  12. Standardized methods and quality control limits for agar and broth microdilution susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum.

    PubMed

    Waites, Ken B; Duffy, Lynn B; Bébéar, Cécile M; Matlow, Anne; Talkington, Deborah F; Kenny, George E; Totten, Patricia A; Bade, Donald J; Zheng, Xiaotian; Davidson, Maureen K; Shortridge, Virginia D; Watts, Jeffrey L; Brown, Steven D

    2012-11-01

    An international multilaboratory collaborative study was conducted to develop standard media and consensus methods for the performance and quality control of antimicrobial susceptibility testing of Mycoplasma pneumoniae, Mycoplasma hominis, and Ureaplasma urealyticum using broth microdilution and agar dilution techniques. A reference strain from the American Type Culture Collection was designated for each species, which was to be used for quality control purposes. Repeat testing of replicate samples of each reference strain by participating laboratories utilizing both methods and different lots of media enabled a 3- to 4-dilution MIC range to be established for drugs in several different classes, including tetracyclines, macrolides, ketolides, lincosamides, and fluoroquinolones. This represents the first multilaboratory collaboration to standardize susceptibility testing methods and to designate quality control parameters to ensure accurate and reliable assay results for mycoplasmas and ureaplasmas that infect humans.

  13. AgarTrap: a simplified Agrobacterium-mediated transformation method for sporelings of the liverwort Marchantia polymorpha L.

    PubMed

    Tsuboyama, Shoko; Kodama, Yutaka

    2014-01-01

    The liverwort Marchantia polymorpha L. is being developed as an emerging model plant, and several transformation techniques were recently reported. Examples are biolistic- and Agrobacterium-mediated transformation methods. Here, we report a simplified method for Agrobacterium-mediated transformation of sporelings, and it is termed Agar-utilized Transformation with Pouring Solutions (AgarTrap). The procedure of the AgarTrap was carried out by simply exchanging appropriate solutions in a Petri dish, and completed within a week, successfully yielding sufficient numbers of independent transformants for molecular analysis (e.g. characterization of gene/protein function) in a single experiment. The AgarTrap method will promote future molecular biological study in M. polymorpha.

  14. Simplified Method for Groundwater Treatment Using Dilution and Ceramic Filter

    NASA Astrophysics Data System (ADS)

    Musa, S.; Ariff, N. A.; Kadir, M. N. Abdul; Denan, F.

    2016-07-01

    Groundwater is one of the natural resources that is not susceptible to pollutants. However, increasing activities of municipal, industrial, agricultural or extreme land use activities have resulted in groundwater contamination as occured at the Research Centre for Soft Soil Malaysia (RECESS), Universiti Tun Hussein Onn Malaysia (UTHM). Thus, aims of this study is to treat groundwater by using rainwater and simple ceramic filter as a treatment agent. The treatment uses rain water dilution, ceramic filters and combined method of dilute and filtering as an alternate treatment which are simple and more practical compared to modern or chemical methods. The water went through dilution treatment processes able to get rid of 57% reduction compared to initial condition. Meanwhile, the water that passes through the filtering process successfully get rid of as much as 86% groundwater parameters where only chloride does not pass the standard. Favorable results for the combination methods of dilution and filtration methods that can succesfully eliminate 100% parameters that donot pass the standards of the Ministry of Health and the Interim National Drinking Water Quality Standard such as those found in groundwater in RECESS, UTHM especially sulfate and chloride. As a result, it allows the raw water that will use clean drinking water and safe. It also proves that the method used in this study is very effective in improving the quality of groundwater.

  15. Comparison of the antibacterial activity of chelating agents using the agar diffusion method

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The agar diffusion assay was used to examine antibacterial activity of 2 metal chelators. Concentrations of 0 to 40 mM of ethylenediaminetetraacetic acid (EDTA) and ethylenediamine-N,N’-disuccinic acid (EDDS) were prepared in 1.0 M potassium hydroxide (KOH). The pH of the solutions was adjusted to 1...

  16. In vitro antifungal susceptibility testing of Scopulariopsis brevicaulis strains using agar diffusion method.

    PubMed

    Skóra, Magdalena; Macura, Anna B

    2011-01-01

    The genus Scopulariopsis is a common soil saprotroph and has been isolated from air, organic waste and also from plant, animal and human tissues. Scopulariopsis has mainly been associated in humans with superficial mycoses, but it has also been described as the cause of subcutaneous and invasive infections. The most common aetiological agent of infections in humans is Scopulariopsis brevicaulis. This species has been reported to be resistant in vitro to broad-spectrum antifungal agents available today. The aim of the study was to establish in vitro antifungal susceptibility of 35 S. brevicaulis strains against amphotericin B (AMB), flucytosine (FC), caspofungin (CAS), terbinafine (TER), ciclopirox (CIC), voriconazole (VOR), clotrimazole (CTR), miconazole (MCZ), econazole (ECO), ketoconazole (KET), itraconazole (ITR), and fluconazole (FLU). Antifungal susceptibility tests were evaluated by an agar diffusion method (Neo-Sensitabs, Rosco, Denmark). AMB, FC, CAS, ITR and FLU showed no antifungal activity against S. brevicaulis. TER, CIC, CTR, KET, VOR, ECO, and MCZ revealed inhibitory activity for S. brevicaulis, but it varied for each of the drugs. The best antifungal effect was observed for TER and CIC. All isolates had large inhibition zones for TER and CIC. CTR was also inhibitory for all tested S. brevicaulis isolates, but the diameters of inhibition zones were smaller than for TER and CIC. Nearly 89% isolates showed inhibition zones for KET and the mean diameter of the inhibition zone was comparable to CTR. The least antifungal activity exhibited VQR, ECO and MCZ. Because of the multiresistance of S. brevicaulis, infections due to this species may not respond to particular antifungal treatment and other therapeutic approaches should be considered, e.g., combined therapy and/or surgery. PMID:21682097

  17. In vitro antifungal susceptibility testing of Scopulariopsis brevicaulis strains using agar diffusion method.

    PubMed

    Skóra, Magdalena; Macura, Anna B

    2011-01-01

    The genus Scopulariopsis is a common soil saprotroph and has been isolated from air, organic waste and also from plant, animal and human tissues. Scopulariopsis has mainly been associated in humans with superficial mycoses, but it has also been described as the cause of subcutaneous and invasive infections. The most common aetiological agent of infections in humans is Scopulariopsis brevicaulis. This species has been reported to be resistant in vitro to broad-spectrum antifungal agents available today. The aim of the study was to establish in vitro antifungal susceptibility of 35 S. brevicaulis strains against amphotericin B (AMB), flucytosine (FC), caspofungin (CAS), terbinafine (TER), ciclopirox (CIC), voriconazole (VOR), clotrimazole (CTR), miconazole (MCZ), econazole (ECO), ketoconazole (KET), itraconazole (ITR), and fluconazole (FLU). Antifungal susceptibility tests were evaluated by an agar diffusion method (Neo-Sensitabs, Rosco, Denmark). AMB, FC, CAS, ITR and FLU showed no antifungal activity against S. brevicaulis. TER, CIC, CTR, KET, VOR, ECO, and MCZ revealed inhibitory activity for S. brevicaulis, but it varied for each of the drugs. The best antifungal effect was observed for TER and CIC. All isolates had large inhibition zones for TER and CIC. CTR was also inhibitory for all tested S. brevicaulis isolates, but the diameters of inhibition zones were smaller than for TER and CIC. Nearly 89% isolates showed inhibition zones for KET and the mean diameter of the inhibition zone was comparable to CTR. The least antifungal activity exhibited VQR, ECO and MCZ. Because of the multiresistance of S. brevicaulis, infections due to this species may not respond to particular antifungal treatment and other therapeutic approaches should be considered, e.g., combined therapy and/or surgery.

  18. A lattice Boltzmann method for dilute polymer solutions.

    PubMed

    Singh, Shiwani; Subramanian, Ganesh; Ansumali, Santosh

    2011-06-13

    We present a lattice Boltzmann approach for the simulation of non-Newtonian fluids. The method is illustrated for the specific case of dilute polymer solutions. With the appropriate local equilibrium distribution, phase-space dynamics on a lattice, driven by a Bhatnagar-Gross-Krook (BGK) relaxation term, leads to a solution of the Fokker-Planck equation governing the probability density of polymer configurations. Results for the bulk rheological characteristics for steady and start-up shear flow are presented, and compare favourably with those obtained using Brownian dynamics simulations. The new method is less expensive than stochastic simulation techniques, particularly in the range of small to moderate Weissenberg numbers (Wi).

  19. Improved method of screening for aflatoxin with a coconut agar medium.

    PubMed

    Davis, N D; Iyer, S K; Diener, U L

    1987-07-01

    Nine isolates of Aspergillus flavus and Aspergillus parasiticus were screened for aflatoxin production on a coconut extract agar medium. Aflatoxin-producing colonies were detected under long-wave UV light (365 nm) by blue fluorescence on the reverse side after 2 to 5 days of growth. Aflatoxin production was verified by chemical analysis. Several types of shredded coconut available in the United States were tested and found to be satisfactory. No additives were required. Various parameters affecting the test were investigated.

  20. Evaluation of PCR-based screening for vancomycin-resistant enterococci compared with a chromogenic agar-based culture method.

    PubMed

    Seo, Ja Young; Kim, Pyung-Whan; Lee, Jang-Ho; Song, Jae-Hoon; Peck, Kyong-Ran; Chung, Doo-Ryeon; Kang, Cheol-In; Ki, Chang-Seok; Lee, Nam Yong

    2011-07-01

    Rapid detection of vancomycin-resistant enterococci (VRE) infection is very important for control and prevention of nosocomial spread of these bacteria. A multiplex PCR method for rapid screening of VRE has recently been developed. We performed a prospective study of VRE screening tests to compare the performance of PCR to that of a chromogenic agar-based culture method. From January to December 2009, a total of 8815 rectal swab specimens were tested simultaneously for VRE by VRE selective culture and by PCR. The specimens were inoculated onto ChromID VRE agar containing 8 µg vancomycin ml⁻¹ and examined after 24 and 48 h of incubation. Identification and antibiotic susceptibility tests were performed using the automated VITEK-2 system and a supplementary E-test and disk diffusion test. Detection of the vanA and vanB genes was performed with the Seeplex VRE detection kit. Specimens were inoculated in enterococcosel broth for 16-24 h before PCR for enrichment of VRE. VRE were isolated from 741 of the 8815 specimens by chromogenic agar-based culture (8.4 %). vanA and vanB genotypes were detected in 758 (8.6 %) and 3 (0.03 %) specimens, respectively, by multiplex PCR. Sensitivity, specificity, positive predictive value and negative predictive value of PCR for detection of VRE were 98.2 %, 99.6 %, 95.7 %, and 99.8 %. No VRE were isolated from vanB-positive specimens. The overall performance of PCR is comparable to that of a chromogenic agar-based culture method for screening of VRE, so PCR could be an alternative or supportive method for effective control of nosocomial VRE infection.

  1. Agar block smear preparation: a novel method of slide preparation for preservation of native fungal structures for microscopic examination and long-term storage.

    PubMed

    Woo, Patrick C Y; Ngan, Antonio H Y; Chui, Hon-Kit; Lau, Susanna K P; Yuen, Kwok-Yung

    2010-09-01

    We describe a novel method of fungal slide preparation named "agar block smear preparation." A total of 510 agar block smears of 25 fungal strains obtained from culture collections, 90 QC fungal strains, and 82 clinical fungal strains from our clinical microbiology laboratory, which included a total of 137 species of yeasts, molds, and thermal dimorphic fungi, were prepared and examined. In contrast to adhesive tape preparation, agar block smears preserved the native fungal structures, such as intact conidiophores of Aspergillus species and arrangements of conidia in Scopulariopsis brevicaulis. Furthermore, agar block smears allowed examination of fungal structures embedded in the agar, such as the ascomata with ascomal hairs in Chaetomium funicola; pycnidium of Phoma glomerata; the intercalary ovoidal chlamydospores arranged in chains of Fusarium dimerum; and the lateral, spherical chlamydospores arranged in pairs of Fusarium solani. After 1 year of storage, morphological integrity was found to have been maintained in 459 (90%) of the 510 agar block smears. After 3 years of storage, morphological integrity was found to have been maintained in 72 (71%) of the 102 smears prepared in 2006. Agar block smear preparation preserves the native fungal structures and allows long-term storage and examination of fungal structures embedded in the agar, hence overcoming the major drawbacks of adhesive tape preparation. The major roles of agar block smear should be diagnosis for difficult cases, accurate identification of fungal species for clinical management of patients and epidemiological studies, and long-term storage for transportation of slides and education purposes.

  2. Radiotracer Dilution Method for Mercury Inventory Study in Electrolytic Cells

    SciTech Connect

    Sugiharto; Su'ud, Zaki; Kurniadi, Rizal; Waris, Abdul; Santoso, Sigit Budi; Abidin, Zainal; Santoso, Gatot Budi

    2010-06-22

    Purpose of the experiment is to demonstrate feasibility the use of radiotracer to measure weight of mercury in electrolytic cells of soda industry. The weight of mercury in each cell of the plant is designed approximately 1700 kg. Radiotracer is prepared by mixing {sup 203}Hg radioactive mercury with 2400 g of inactive mercury in a bath. The respective precisely weighted mercury aliquots to be injected into the cells are prepared by pouring approximately 130 g of radioactive mercury taken from the bath into 13 standard vials, in accordance with the number of the cells tested. Four standard references prepared by further dilution of {+-}2 g active mercury taken from the bath to obtain the dilution factors range of 12,000 to 20,000 from which the calibration graph is constructed. The injection process is conducting by pouring the radioactive mercury from aliquots into the flowing mercury at the inlet side of the cell and allows them to mix thoroughly. It is assumed that the mass of the radiotracer injected into a closed system remains constant, at least during the period of the test. From this experiment it was observed that the mixing time is two days after injection of radioactive mercury. The inactive mercury in each electrolytic cell calculated by the radiotracer method is of the range 1351.529 kg to 1966.354 kg with maximum error (95% confidence) is 1.52 %. The accuracy of measurement of the present method is better than gravimetric one which accounts 4 % of error on average.

  3. An inline QC method for determining serial dilution performance of DMSO-based systems.

    PubMed

    Walling, Leslie A

    2011-06-01

    Serial dilution of compounds solubilized in dimethylsulfoxide (DMSO) for dose-response curves is a common method for efficacy analysis of potential drug candidates. In general, serial dilution methods are particularly prone to error propagation because each dilution is dependent on the previous concentration. Moreover, assumptions about quality control parameters (i.e., dye linearity) can lead to an erroneous process. Here, an inline performance measurement is sought to improve the precision and accuracy of dilution plates. Sulforhodamine 101 (S101) dye is introduced as the quantitative fluorometric method of choice for DMSO-based systems. Although S101 in DMSO behaves in a nonlinear fashion over its detectable range, we account for this with a direct calibration method that includes every point of the dilution template. This report contains dye selection rationale for the S101 dye and its use in quantifying the performance of 96- and 384-well dilution protocols as tested on five identical instruments.

  4. Detection of intermediately vancomycin-susceptible and heterogeneous Staphylococcus aureus isolates: comparison of Etest and Agar screening methods.

    PubMed

    Riederer, K; Shemes, S; Chase, P; Musta, A; Mar, A; Khatib, R

    2011-06-01

    Detection of Staphylococcus aureus isolates with intermediate vancomycin susceptibility (VISA) and heteroresistance (hVISA) remains problematic. The population analysis profile/area under the curve (PAP/AUC) is the gold standard but is cumbersome. We compared the performance of two Etest screening methods (macromethod [MAC] and glycopeptide resistance detection [GRD]) plus brain heart infusion (BHI) agars supplemented with 3 (BHI-V3) or 4 (BHI-V4) mg/liter vancomycin in detecting hVISA and/or VISA phenotypes. Etest hVISA screenings were done in parallel for 485 saved methicillin-resistant S. aureus (MRSA) blood isolates according to the manufacturer's instructions. The PAP/AUC was measured for all isolates according to the modified method. PAP/AUC test isolate/Mu3 ratios of <0.9, 0.9 to 1.3, and >1.3 were considered positive for susceptible MRSA (S-MRSA), hVISA, and VISA, respectively. PAP/AUC revealed seven VISA and 33 hVISA phenotypes. MAC screening was positive for 30 (75.0%) hVISA/VISA and 49 (11.0%) S-MRSA isolates. GRD screening was positive for 28 (70.0%) hVISA/VISA and 63 (14.2%) S-MRSA isolates. Growth on BHI-V3 was noted in all hVISA/VISA and 24 (5.4%) S-MRSA isolates. Growth on BHI-V4 was noted in all VISA and four (12.1%) hVISA isolates. None of the S-MRSA isolates grew on BHI-V4 agar. The sensitivity, specificity, and positive (PPV) and negative (NPV) predictive values were 75.0%, 89.0%, 38.0%, and 97.5% for MAC; 70.0%, 85.8%, 30.8%, and 97.0% for GRD; 100%, 94.6%, 62.5%, and 100% for BHI-V3; and 100, 99.2%, 63.6%, and 100% for BHI-V4 (for detecting VISA). These findings suggest that both Etest screening methods have excellent NPV, but positive results require confirmation. BHI-V3 and BHI-V4 agars provide more precise identification of hVISA and VISA, respectively; they may be reasonable alternatives to PAP/AUC.

  5. The agar diffusion scratch assay - A novel method to assess the bioactive and cytotoxic potential of new materials and compounds

    PubMed Central

    Pusnik, Mascha; Imeri, Minire; Deppierraz, Grégoire; Bruinink, Arie; Zinn, Manfred

    2016-01-01

    A profound in vitro evaluation not only of the cytotoxic but also of bioactive potential of a given compound or material is crucial for predicting potential effects in the in vivo situation. However, most of the current methods have weaknesses in either the quantitative or qualitative assessment of cytotoxicity and/or bioactivity of the test compound. Here we describe a novel assay combining the ISO 10993-5 agar diffusion test and the scratch also termed wound healing assay. In contrast to these original tests this assay is able to detect and distinguish between cytotoxic, cell migration modifying and cytotoxic plus cell migration modifying compounds, and this at higher sensitivity and in a quantitative way. PMID:26861591

  6. Quantifying Methane Fluxes Simply and Accurately: The Tracer Dilution Method

    NASA Astrophysics Data System (ADS)

    Rella, Christopher; Crosson, Eric; Green, Roger; Hater, Gary; Dayton, Dave; Lafleur, Rick; Merrill, Ray; Tan, Sze; Thoma, Eben

    2010-05-01

    Methane is an important atmospheric constituent with a wide variety of sources, both natural and anthropogenic, including wetlands and other water bodies, permafrost, farms, landfills, and areas with significant petrochemical exploration, drilling, transport, or processing, or refining occurs. Despite its importance to the carbon cycle, its significant impact as a greenhouse gas, and its ubiquity in modern life as a source of energy, its sources and sinks in marine and terrestrial ecosystems are only poorly understood. This is largely because high quality, quantitative measurements of methane fluxes in these different environments have not been available, due both to the lack of robust field-deployable instrumentation as well as to the fact that most significant sources of methane extend over large areas (from 10's to 1,000,000's of square meters) and are heterogeneous emitters - i.e., the methane is not emitted evenly over the area in question. Quantifying the total methane emissions from such sources becomes a tremendous challenge, compounded by the fact that atmospheric transport from emission point to detection point can be highly variable. In this presentation we describe a robust, accurate, and easy-to-deploy technique called the tracer dilution method, in which a known gas (such as acetylene, nitrous oxide, or sulfur hexafluoride) is released in the same vicinity of the methane emissions. Measurements of methane and the tracer gas are then made downwind of the release point, in the so-called far-field, where the area of methane emissions cannot be distinguished from a point source (i.e., the two gas plumes are well-mixed). In this regime, the methane emissions are given by the ratio of the two measured concentrations, multiplied by the known tracer emission rate. The challenges associated with atmospheric variability and heterogeneous methane emissions are handled automatically by the transport and dispersion of the tracer. We present detailed methane flux

  7. Estimating dilutions for patch testing skin care products: a practical method.

    PubMed

    Sherertz, E F; Byers, S V

    1997-09-01

    When patch testing, it is helpful to patch test with the patient's own topical products. However, the thickness (viscosity) of the product often prevents easy measurement for dilution. A method of estimating volume: volume dilutions that requires minimal investment in supplies is presented. This method is only applicable to personal products and is not suitable for industrial or household chemicals.

  8. The antibacterial activity of sodium hypochlorite and chlorhexidine against Enterococcus faecalis: A review on agar diffusion and direct contact methods

    PubMed Central

    Luddin, Norhayati; Ahmed, Hany Mohamed Aly

    2013-01-01

    Complete debridement and disinfection of the root canal system are fundamental requirements for successful endodontic treatment. Despite the morphological challenges of the internal root anatomy, root canal irrigants play an important role in the optimization of the root canal preparation, which is essentially a chemo-mechanical procedure. Enterococcus faecalis is one of the most resistant microorganisms that dominants the microbial ecosystem of persistent periradicular lesions in retreatment cases. For that reason, many in vitro and in vivo studies evaluated and compared the antibacterial activity of sodium hypochlorite and chlorhexidine at varying concentrations using different experimental models against this microorganism. However, many controversies with regard to the ideal irrigant and concentration do in fact exist. Hence, this review aims to discuss the antibacterial activity of these two main root canal irrigants against Enterococcus faecalis using the agar diffusion and direct contact methods and the possible modulating factors responsible for inconsistent findings among different studies. In addition, the disinfection potential of both chemical agents on gutta percha and Resilon cones are also discussed. The source of this review was conducted through an electronic literature search using PubMed database from December 1997 until December 2011, which analyze the related laboratory investigations of both irrigants, published in major endodontic journals. PMID:23349569

  9. An Evaluation Method for the Suppression of Pathogenic Fusarium oxysporum by Soil Microorganisms Using the Dilution Plate Technique.

    PubMed

    Mitsuboshi, Masahiro; Kioka, Yuuzou; Noguchi, Katsunori; Asakawa, Susumu

    2016-09-29

    Soil-borne diseases caused by pathogenic microorganisms are one of the main factors responsible for the decline in crop yields in farmlands. Pathogenic Fusarium oxysporum causes serious damage to various crops, and, thus, a feasible diagnostic method for soil-borne diseases is required. We herein examined a simple method to evaluate the suppressiveness of soil microorganisms against a pathogen by co-cultivating indigenous soil microorganisms and a pathogenic fungus (F. oxysporum f. sp. spinaciae). We inoculated F. oxysporum onto the center of agar medium plates mixed with a dilution series of a suspension of organic fertilizers or soil. After an approximately one-week cultivation, the growth degree of F. oxysporum was estimated based on the size of the colonies that formed on the plates. The growth degree of F. oxysporum significantly differed among the organic fertilizers tested, indicating the usefulness of the method for evaluating suppressiveness by organic fertilizers. Differences in the growth degrees of F. oxysporum were associated with the incidence of disease in spinach on soil treated with organic fertilizers and inoculated with a pathogenic F. oxysporum strain. These results suggested that this method provides some useful information on the suppressiveness of organic fertilizers and soil against Fusarium wilt. PMID:27558588

  10. An Evaluation Method for the Suppression of Pathogenic Fusarium oxysporum by Soil Microorganisms Using the Dilution Plate Technique

    PubMed Central

    Mitsuboshi, Masahiro; Kioka, Yuuzou; Noguchi, Katsunori; Asakawa, Susumu

    2016-01-01

    Soil-borne diseases caused by pathogenic microorganisms are one of the main factors responsible for the decline in crop yields in farmlands. Pathogenic Fusarium oxysporum causes serious damage to various crops, and, thus, a feasible diagnostic method for soil-borne diseases is required. We herein examined a simple method to evaluate the suppressiveness of soil microorganisms against a pathogen by co-cultivating indigenous soil microorganisms and a pathogenic fungus (F. oxysporum f. sp. spinaciae). We inoculated F. oxysporum onto the center of agar medium plates mixed with a dilution series of a suspension of organic fertilizers or soil. After an approximately one-week cultivation, the growth degree of F. oxysporum was estimated based on the size of the colonies that formed on the plates. The growth degree of F. oxysporum significantly differed among the organic fertilizers tested, indicating the usefulness of the method for evaluating suppressiveness by organic fertilizers. Differences in the growth degrees of F. oxysporum were associated with the incidence of disease in spinach on soil treated with organic fertilizers and inoculated with a pathogenic F. oxysporum strain. These results suggested that this method provides some useful information on the suppressiveness of organic fertilizers and soil against Fusarium wilt. PMID:27558588

  11. Comparative evaluation of a chromogenic agar medium-PCR protocol with a conventional method for isolation of Vibrio parahaemolyticus strains from environmental and clinical samples.

    PubMed

    Canizalez-Roman, Adrian; Flores-Villaseñor, Héctor; Zazueta-Beltran, Jorge; Muro-Amador, Secundino; León-Sicairos, Nidia

    2011-02-01

    Screening for pathogenic Vibrio parahaemolyticus has become routine in certain areas associated with food-borne outbreaks. This study is an evaluation of the CHROMagar Vibrio (CV) medium-PCR protocol and the conventional method (TCBS (thiosulfate-citrate-bile salts-sucrose) agar plus biochemical and Wagatsuma agar tests) for detection of V. parahaemolyticus in shrimp, water, sediment, and stool samples collected for biosurveillance in an endemic area of northwestern Mexico. A total of 131 environmental and clinical samples were evaluated. The CV medium-PCR protocol showed a significantly improved ability (P < 0.05) to isolate and detect V. parahaemolyticus, identifying isolates of this bacteria missed by the conventional method. Although some other bacteria, distinct from pathogenic V. parahaemolyticus, produced violet colonies similar to that of V. parahaemolyticus on CV medium, we were able to detect a superior number of samples of V. parahaemolyticus with the CV medium-PCR protocol than with the conventional method. The Kanagawa phenomenon is routinely determined on Wagatsuma agar for the diagnosis of V. parahaemolyticus (pathogenic) positive for thermostable direct hemolysin (TDH) in developing countries. In our results, Wagatsuma agar showed low sensitivity (65.4% at 24 h and 75.6% at 48 h) and specificity (52.4% at 48 h) for identifying V. parahaemolyticus positive for TDH. Overall, our data support the use of the CV medium-PCR protocol in place of the conventional method (TCBS-biochemical tests-Wagatsuma agar) for detection of pathogenic V. parahaemolyticus, both in terms of effectiveness and cost efficiency.

  12. Comparison of Results Obtained by Testing with Three Different Agar Media and by the NCCLS M27-A Method for In Vitro Testing of Fluconazole against Candida spp.

    PubMed Central

    Rubio, M. Carmen; Gil, Joaquina; de Ocáriz, Inmaculada Ramírez; Benito, Rafael; Rezusta, Antonio

    2003-01-01

    Fluconazole susceptibilities of 150 Candida isolates were determined by a 25-μg fluconazole disk diffusion agar test and compared with the microdilution NCCLS M27-A method. The agar test used three different media and was read at 24 and 48 h. When only the susceptible and nonsusceptible categories were used, disk diffusion with Müeller-Hinton agar supplemented with 2% glucose and 0.5 μg of methylene blue (MHGM) per ml had a 95.37% correlation with the MIC method at 24 h, followed by RPMI 1640-2% of glucose agar (correlation, 94%) and Shadomy medium (SHDM) (correlation, 92.6%). The growth of microcolonies inside the inhibition zones was common (>63%) in the RPMI and SHDM media and minimal with MHGM (8.7%). At 48 h, MHGM and SHDM still had a >91% correlation with the MIC, while RPMI results had dropped to 75%. The best overall agreement was obtained with C. dubliniensis (100%). PMID:12791899

  13. [An observation on the histological structure of Oncomelania hupensis soft tissue by agar-paraffin double-embedding method].

    PubMed

    Tan, Ping; Zhang, Jie; Li, Qing; Yu, Zhi-jun

    2014-12-01

    In order to study the histological structure of Oncomelania hupensis soft tissue, the fixed soft tissues of O. hupensis were pre-embedded in the agar and made blocks, then dehydrated, transparentized, immersed in paraffin, sectioned, and stained with haematoxylin-eosin (HE). Permanent slides of O. hupensis soft tissue were obtained. The histological structure of soft tissues was clear under the microscope.

  14. 21 CFR 582.7115 - Agar-agar.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Agar-agar. 582.7115 Section 582.7115 Food and..., FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7115 Agar-agar. (a) Product. Agar-agar. (b) Conditions of use. This substance is generally recognized as safe when used...

  15. 21 CFR 582.7115 - Agar-agar.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 6 2011-04-01 2011-04-01 false Agar-agar. 582.7115 Section 582.7115 Food and..., FEEDS, AND RELATED PRODUCTS SUBSTANCES GENERALLY RECOGNIZED AS SAFE Stabilizers § 582.7115 Agar-agar. (a) Product. Agar-agar. (b) Conditions of use. This substance is generally recognized as safe when used...

  16. A novel agar formulation for isolation and direct enumeration of Vibrio vulnificus from oyster tissue.

    PubMed

    Griffitt, Kimberly J; Grimes, D Jay

    2013-08-01

    A new selective and differential medium, Vibrio vulnificus X-Gal (VVX), was developed for direct enumeration of V. vulnificus (Vv) from oyster samples. This agar utilizes cellobiose and lactose as carbon sources, and the antibiotics colistin and polymyxin B as selective agents. Hydrolysis of 5-bromo-4-chloro-3-indolyl- beta-d-galactopyranoside (x-gal), used in the agar as a lactose analog, produces an insoluble blue dye that makes lactose positive colonies easily distinguishable from any non-lactose fermenting bacteria. Various bacterial species were spot plated onto thiosulfate-citrate-bile salts-sucrose agar (TCBS), and CHROMagar Vibrio, two vibrio-specific selective agars, non-selective agar, and VVX to compare selectivity of VVX to other widely used media. A V. vulnificus pure culture was serially diluted on VVX and non-selective agar to determine the VVX percent recovery. Water and oyster samples were spread plated on VVX agar and allowed to incubate for 16-18 h at 33 °C. Blue and white colonies from VVX agar were picked and screened by end point PCR for the Vv hemolysin vvhA. VVX agar showed a significant improvement over TCBS and CHROMagar at preventing non-target growth. There was an 87.5% recovery compared to non-selective plating and a 98% positivity rate of blue colonies picked from oyster tissue plating. The findings suggest that this new agar is a fast, distinctive, and accurate method for enumeration of V. vulnificus from the environment.

  17. Comparison of a novel MPN method against the yeast extract agar (YEA) pour plate method for the enumeration of heterotrophic bacteria from drinking water.

    PubMed

    Sartory, David P; Gu, Haoyi; Chen, Chun-Ming

    2008-07-01

    This study compared the Quanti-Disc most probable number (MPN) test for heterotrophic bacteria from drinking water with the widely used yeast extract agar (YEA) pour plate method. The Quanti-Disc test module contains 50 reaction wells in which a medium has been pre-deposited. The medium contains a suite of three fluorogenic enzyme substrates selected for the detection of enzymes expressed widely by heterotrophic bacteria. The MPN of heterotrophic bacteria is calculated from the number of fluorescing reaction wells after incubation of a sample. Quanti-Disc and the YEA pour plate method were compared according to guidance on comparing methods given in United Kingdom national guidance and ISO 17994:2004. The two methods were also challenged with reference strains and isolates of heterotrophic bacteria from drinking water. This indicated that heterotrophic bacteria commonly encountered in drinking water are detected by both the YEA pour plate method and Quanti-Disc. Analysis of data from split water samples (723 for 37 degrees C tests and 872 for 22 degrees C tests) from nine geographically diverse laboratories in England and Wales demonstrated that the Quanti-Disc method is equivalent to the YEA pour plate method for the analysis of heterotrophic bacteria from drinking and similar waters at 37 degrees C, and superior to YEA for the analysis at 22 degrees C. The Quanti-Disc method is a simple and efficient alternative method for the enumeration of heterotrophic bacteria from drinking water.

  18. Comparison of Sorbitol MacConkey Agar and a Two-Step Method Which Utilizes Enzyme-Linked Immunosorbent Assay Toxin Testing and a Chromogenic Agar To Detect and Isolate Enterohemorrhagic Escherichia coli

    PubMed Central

    Novicki, Thomas J.; Daly, Judy A.; Mottice, Susan L.; Carroll, Karen C.

    2000-01-01

    Enterohemorrhagic Escherichia coli (EHEC) and specifically serotype O157:H7 are a significant cause of hemorrhagic gastrointestinal disease and the hemolytic uremic syndrome. Methods currently used in clinical microbiology labs, such as sorbitol-MacConkey (SMAC) agar, reliably detect only O157:H7. We have evaluated a two-step method that has the potential to identify and isolate all EHEC serotypes, including serotype O157:H7. This method utilizes a chromogenic selective-differential medium for the isolation of E. coli together with an enzyme-linked immunosorbent assay (ELISA) that detects the Shiga-like toxins Stx1 and Stx2. Both are commercially available and usable in a wide range of clinical microbiology laboratories. Compared to a Vero cell cytotoxic assay, SMAC had sensitivities of 23.5% for the identification of all EHEC serotypes and of 50.0% for the identification of O157:H7 alone. The two-step method had sensitivities of 76.5 and 100%, respectively. The ELISA alone had a sensitivity of 82.4% in the detection of Stx1 and Stx2. The specificity was 100% in all cases. Overall, 14 EHEC isolates were obtained: 8 (58%) O157:H7, 2 (14%) O26, 2 (14%) O111:NM, 1 (7%) O103:H2, and 1 (7%) O121:H19. All but one were isolated during the months of May to September. The two-step method was found to be considerably more expensive than SMAC for both positive and negative samples. PMID:10655343

  19. 21 CFR 184.1115 - Agar-agar.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 3 2011-04-01 2011-04-01 false Agar-agar. 184.1115 Section 184.1115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1115 Agar-agar. (a) Agar-agar (CAS Reg. No. PM 9002-18-0) is a...

  20. 21 CFR 184.1115 - Agar-agar.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Agar-agar. 184.1115 Section 184.1115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN... Substances Affirmed as GRAS § 184.1115 Agar-agar. (a) Agar-agar (CAS Reg. No. PM 9002-18-0) is a...

  1. A simplified semiquantitative determination of hepatitis C virus genome molecules by the end-point dilution method.

    PubMed

    Bathelier, C; Mercier, G; Lucotte, G

    1996-12-01

    We describe a semiquantitative method to measure hepatitis C virus (HCV) viral particle numbers, by carrying out reverse-transcription polymerase chain reaction (RT-PCR) on serial dilutions of serum samples. The virus concentrations measured were 10(3)-10(6) viral particles ml-1 of serum. The method described is relatively quick, and the only required manipulation is dilution of the serum. An optimal RT-PCR method is used for diluted and undiluted samples.

  2. Body composition among Sri Lankan infants by 18*O dilution method and the validity of anthropometric equations to predict body fat against 18*O dilution

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Body composition indicators provide a better guidance for growth and nutritional status of the infants. This study was designed to (1) measure the body composition of the Sri Lankan infants using a reference method, the 18*O dilution method; (2) calculate the body fat content of the infants using pu...

  3. Numerical simulations to assess the tracer dilution method for measurement of landfill methane emissions.

    PubMed

    Taylor, Diane M; Chow, Fotini K; Delkash, Madjid; Imhoff, Paul T

    2016-10-01

    Landfills are a significant contributor to anthropogenic methane emissions, but measuring these emissions can be challenging. This work uses numerical simulations to assess the accuracy of the tracer dilution method, which is used to estimate landfill emissions. Atmospheric dispersion simulations with the Weather Research and Forecast model (WRF) are run over Sandtown Landfill in Delaware, USA, using observation data to validate the meteorological model output. A steady landfill methane emissions rate is used in the model, and methane and tracer gas concentrations are collected along various transects downwind from the landfill for use in the tracer dilution method. The calculated methane emissions are compared to the methane emissions rate used in the model to find the percent error of the tracer dilution method for each simulation. The roles of different factors are examined: measurement distance from the landfill, transect angle relative to the wind direction, speed of the transect vehicle, tracer placement relative to the hot spot of methane emissions, complexity of topography, and wind direction. Results show that percent error generally decreases with distance from the landfill, where the tracer and methane plumes become well mixed. Tracer placement has the largest effect on percent error, and topography and wind direction both have significant effects, with measurement errors ranging from -12% to 42% over all simulations. Transect angle and transect speed have small to negligible effects on the accuracy of the tracer dilution method. These tracer dilution method simulations provide insight into measurement errors that might occur in the field, enhance understanding of the method's limitations, and aid interpretation of field data. PMID:27395754

  4. Numerical simulations to assess the tracer dilution method for measurement of landfill methane emissions.

    PubMed

    Taylor, Diane M; Chow, Fotini K; Delkash, Madjid; Imhoff, Paul T

    2016-10-01

    Landfills are a significant contributor to anthropogenic methane emissions, but measuring these emissions can be challenging. This work uses numerical simulations to assess the accuracy of the tracer dilution method, which is used to estimate landfill emissions. Atmospheric dispersion simulations with the Weather Research and Forecast model (WRF) are run over Sandtown Landfill in Delaware, USA, using observation data to validate the meteorological model output. A steady landfill methane emissions rate is used in the model, and methane and tracer gas concentrations are collected along various transects downwind from the landfill for use in the tracer dilution method. The calculated methane emissions are compared to the methane emissions rate used in the model to find the percent error of the tracer dilution method for each simulation. The roles of different factors are examined: measurement distance from the landfill, transect angle relative to the wind direction, speed of the transect vehicle, tracer placement relative to the hot spot of methane emissions, complexity of topography, and wind direction. Results show that percent error generally decreases with distance from the landfill, where the tracer and methane plumes become well mixed. Tracer placement has the largest effect on percent error, and topography and wind direction both have significant effects, with measurement errors ranging from -12% to 42% over all simulations. Transect angle and transect speed have small to negligible effects on the accuracy of the tracer dilution method. These tracer dilution method simulations provide insight into measurement errors that might occur in the field, enhance understanding of the method's limitations, and aid interpretation of field data.

  5. Assessing the Accuracy of the Tracer Dilution Method with Atmospheric Dispersion Modeling

    NASA Astrophysics Data System (ADS)

    Taylor, D.; Delkash, M.; Chow, F. K.; Imhoff, P. T.

    2015-12-01

    Landfill methane emissions are difficult to estimate due to limited observations and data uncertainty. The mobile tracer dilution method is a widely used and cost-effective approach for predicting landfill methane emissions. The method uses a tracer gas released on the surface of the landfill and measures the concentrations of both methane and the tracer gas downwind. Mobile measurements are conducted with a gas analyzer mounted on a vehicle to capture transects of both gas plumes. The idea behind the method is that if the measurements are performed far enough downwind, the methane plume from the large area source of the landfill and the tracer plume from a small number of point sources will be sufficiently well-mixed to behave similarly, and the ratio between the concentrations will be a good estimate of the ratio between the two emissions rates. The mobile tracer dilution method is sensitive to different factors of the setup such as placement of the tracer release locations and distance from the landfill to the downwind measurements, which have not been thoroughly examined. In this study, numerical modeling is used as an alternative to field measurements to study the sensitivity of the tracer dilution method and provide estimates of measurement accuracy. Using topography and wind conditions for an actual landfill, a landfill emissions rate is prescribed in the model and compared against the emissions rate predicted by application of the tracer dilution method. Two different methane emissions scenarios are simulated: homogeneous emissions over the entire surface of the landfill, and heterogeneous emissions with a hot spot containing 80% of the total emissions where the daily cover area is located. Numerical modeling of the tracer dilution method is a useful tool for evaluating the method without having the expense and labor commitment of multiple field campaigns. Factors tested include number of tracers, distance between tracers, distance from landfill to transect

  6. A Dilute-and-Shoot LC-MS Method for Quantitating Opioids in Oral Fluid.

    PubMed

    Enders, Jeffrey R; McIntire, Gregory L

    2015-10-01

    Opioid testing represents a dominant share of the market in pain management clinical testing facilities. Testing of this drug class in oral fluid (OF) has begun to rise in popularity. OF analysis has traditionally required extensive clean-up protocols and sample concentration, which can be avoided. This work highlights the use of a fast, 'dilute-and-shoot' method that performs no considerable sample manipulation. A quantitative method for the determination of eight common opioids and associated metabolites (codeine, morphine, hydrocodone, hydromorphone, norhydrocodone, oxycodone, noroxycodone and oxymorphone) in OF is described herein. OF sample is diluted 10-fold in methanol/water and then analyzed using an Agilent chromatographic stack coupled with an AB SCIEX 4500. The method has a 2.2-min LC gradient and a cycle time of 2.9 min. In contrast to most published methods of this particular type, this method uses no sample clean-up or concentration and has a considerably faster LC gradient, making it ideal for very high-throughput laboratories. Importantly, the method requires only 100 μL of sample and is diluted 10-fold prior to injection to help with instrument viability. Baseline separation of all isobaric opioids listed above was achieved on a phenyl-hexyl column. The validated calibration range for this method is 2.5-1,000 ng/mL. This 'dilute-and-shoot' method removes the unnecessary, costly and time-consuming extraction steps found in traditional methods and still surpasses all analytical requirements. PMID:26378142

  7. Determination of beryllium in ores and rocks by a dilution-fluorometric method with morin

    USGS Publications Warehouse

    May, R.; Grimaldi, F.S.

    1961-01-01

    Beryllium in concentrations as little as a few parts per million is determined fluorometrically with morin in low grade ores by a dilution method without separations. A high sensitivity is obtained by the adoption of instrumental and reaction conditions that give a satisfactory ratio of beryllium to blank fluorescence and at the same time minimize iron interference. Data on the behavior of 47 ions are given. The method is applied to ores containing bertrandite and beryl as the beryllium minerals.

  8. Bacteria holding times for fecal coliform by mFC agar method and total coliform and Escherichia coli by Colilert®-18 Quanti-Tray® method

    USGS Publications Warehouse

    Aulenbach, Brent T.

    2010-01-01

    Bacteria holding-time experiments of up to 62 h were performed on five surface-water samples from four urban stream sites in the vicinity of Atlanta, GA, USA that had relatively high densities of coliform bacteria (Escherichia coli densities were all well above the US Environmental Protection Agency criterion of 126 colonies (100 ml) − 1 for recreational waters). Holding-time experiments were done for fecal coliform using the membrane filtration modified fecal coliform (mFC) agar method and for total coliform and E. coli using the Colilert®-18 Quanti-Tray® method. The precisions of these analytical methods were quantified. Precisions determined for fecal coliform indicated that the upper bound of the ideal range of counts could reasonably be extended upward and would improve precision. For the Colilert®-18 method, analytical precisions were similar to the theoretical precisions for this method. Fecal and total coliform densities did not change significantly with holding times up to about 27 h. Limited information indicated that fecal coliform densities might be stable for holding times of up to 62 h, whereas total coliform densities might not be stable for holding times greater than about 27 h. E. coli densities were stable for holding times of up to 18 h—a shorter period than indicated from a previous studies. These results should be applicable to non-regulatory monitoring sampling designs for similar urban surface-water sample types.

  9. Pleural liquid clearance rate measured in awake sheep by the volume of dilution method

    SciTech Connect

    Broaddus, V.C.; Wiener-Kronish, J.P.; Berthiaume, Y.; Staub, N.C.

    1986-03-01

    The authors reported 24h clearance of mock pleural effusions measured terminally in sheep. To measure effusion volume at different times in the same sheep, they injected /sup 111/In-transferrin and measured its dilution. In 5 sheep with effusions of known sizes, the method was accurate to +/-10%. In 5 awake sheep, the authors injected 10 ml/kg of a 1% protein solution via a non-penetrating rib capsule. At 6h, the authors measured the volume by the dilution method and at 24h by direct recovery. The clearance rate in each animal was constant at 2.9-6.0%/h (average 4.8 +/- 1.3%/h). This new method gives a reliable two point clearance rate and requires fewer animals.

  10. Xanthan gum: an economical partial substitute for agar in microbial culture media.

    PubMed

    Babbar, Shashi B; Jain, Ruchi

    2006-04-01

    Xanthan gum, microbial desiccation-resistant polysaccharide prepared commercially by aerobic submerged fermentation from Xanthomonas campestris, has been successfully used alone and in combination with agar for microbial culture media. As illustrative examples, eight bacteria and eight fungi were grown on media solidified with either agar (A, 1.5%), xanthan gum (X, 1%), or combinations of both (0.9% X + 0.1% A, 0.8% X + 0.2% A, 0.7% X + 0.3% A, 0.6% X + 0.4% A). All fungi and bacteria exhibited normal growth and differentiation in all these treatments. Rather, growth of most of the fungi was better on xanthan (alone) and xanthan + agar media than agar medium. As the media gelled with xanthan gum alone flow, it was not possible to incubate Petri plates in inverted position. Moreover, because of the softness, streaking of bacteria was difficult on such media. However, these problems could be overcome by partially replacing xanthan gum with 0.3% agar. Bacterial enumeration studies carried out for Serratia sp. and Pseudomonas sp. by serial dilution and pour-plate method on agar (1.5%), 0.7%/0.6% X + 0.3%/0.4% A yielded similar counts. Selective media, succinate medium for Pseudomonas sp., and MacConkey broth medium for Escherichia coli gelled with 0.7%/0.6% X + 0.3%/0.4% A did not support growth of other bacteria when inoculated along with the above-mentioned bacteria. Likewise, differential medium, CRMA (Congo red mannitol agar) gelled with xanthan-agar combination could differentiate between Agrobacterium tumefaciens and Rhizobium sp.

  11. Reverse isotope dilution method for determining benzene and metabolites in tissues

    SciTech Connect

    Bechtold, W.E.; Sabourin, P.J.; Henderson, R.F.

    1988-07-01

    A method utilizing reverse isotope dilution for the analysis of benzene and its organic soluble metabolites in tissues of rats and mice is presented. Tissues from rats and mice that had been exposed to radiolabeled benzene were extracted with ethyl acetate containing known, excess quantities of unlabeled benzene and metabolites. Butylated hydroxytoluene was added as an antioxidant. The ethyl acetate extracts were analyzed with semipreparative reversed-phase HPLC. Isolated peaks were collected and analyzed for radioactivity (by liquid scintillation spectrometry) and for mass (by UV absorption). The total amount of each compound present was calculated from the mass dilution of the radiolabeled isotope. This method has the advantages of high sensitivity, because of the high specific activity of benzene, and relative stability of the analyses, because of the addition of large amounts of unlabeled carrier analogue.

  12. Isotopic Dilution GC/MS Method for Methionine Determination in Biological Media

    NASA Astrophysics Data System (ADS)

    Horj, Elena; Iordache, Andreea; Culea, Monica

    2011-10-01

    The isotopic dilution mass spectrometry technique is the method of choice for sensitive and accurate determination of analytes in biological samples. The aim of this work was to establish a sensitive analytical method for the determination of methionine in different biological media. Quantitation of methionine from the resultant tracer spectrum requires deconvolution of the enrichment of the isotopomers. Deconvolution of the ion abundance ratios to yield tracer-to-tracee ratio for the isotopomer was done using Brauman's least squares approach. Comparison with regression curve calculation method is presented. The method was applied for amino-acids determination in beef, pork and fish meat.

  13. Melanin bleaching with dilute hydrogen peroxide: a simple and rapid method.

    PubMed

    Liu, Chia-Hsing; Lin, Chih-Hung; Tsai, Min-Jan; Chen, Wan-Tzu; Chai, Chee-Yin; Huang, Ya-Chun; Tsai, Kun-Bow

    2013-05-01

    Melanins are naturally occurring pigments in both normal and pathologic tissues. Two common bleaching processes are potassium permanganate followed by oxalic acid treatment and dilute hydrogen peroxide (H2O2) process. The potassium permanganate/oxalic acid method is faster and more easily incorporated in conventional daily immunostaining protocols, whereas the dilute H2O2 method requires 24 hours. This study aimed to reduce melanin bleaching time by using a 10% H2O2 dilution. First, reaction time was reduced to 30 minutes by raising the temperature to 65°C. Second, containers with high thermal conductivity were used to improve bleaching effectiveness. Experimental comparisons of melanin treatments with H2O2 contained in an iron jar, a glass coplin jar, and a plastic steel jar obtained bleaching time of 20, 30, and 40 minutes, respectively. These modifications of the conventional bleaching method significantly improve the speed and efficiency of the procedure and are recommended when performing immunohistochemical studies.

  14. Turbidity as a method of preparing sperm dilutions in the echinoid sperm/egg bioassay

    SciTech Connect

    Hall, T.J.; Haley, R.K.; Battan, K.J. )

    1993-11-01

    The use of turbidimeter for preparing sperm dilutions used in the echinoid sperm/egg bioassay was evaluated. Regression analyses of the relationship between sperm density and turbidity for the sea urchins Strongylocentrotus purpuratus and Strongylocentrotus droebachiensis and the sand dollar Dendraster excentricus indicated that although there were slope differences for each species, each coefficient of determination was highly significant. For Dendraster excentricus, triplicate hemacytometer counts over a range of turbidities as well as repeated preparations of a single sperm turbidity indicated similar variability for each. The use of the turbidimeter has time-saving advantages over conventional hemacytometer methods without sacrificing precision. Sperm dilutions can be prepared rapidly, minimizing seawater sperm preactivation before test initiation, and may therefore contribute to increased test precision.

  15. Comparison of recovery of airborne microorganisms in a dairy cattle facility using selective agar and thin agar layer resuscitation media.

    PubMed

    Crozier-Dodson, Beth Ann; Fung, Daniel Y C

    2002-09-01

    Thin agar layer (TAL) medium was developed at Kansas State University to improve the resuscitation of injured cells and has been shown to result in higher recovery than is obtained with selective media alone for cold-, heat-, salt-, and acid-injured cells. The experiment presented here was designed to determine the effectiveness of the TAL method for the recovery of possibly injured organisms from air. Eleven agar media were used for the experiment: tryptic soy agar (TSA), MacConkey sorbitol agar (MSA), TAL-MSA, Baird-Parker (BP) agar, TAL-BP agar, modified Oxford (MOX) agar, TAL-MOX agar, xylose lysine sodium desoxycholate (XLD) agar, TAL-XLD agar, Yersinia-selective (CIN) agar, and TAL-CIN agar. The TAL plates were prepared by pipetting 6 ml of selective agar into a BBL Rodac plate (65 by 15 mm). Selective agar was allowed to solidify, and then each plate was overlaid with 6 ml of TSA. Selective agar plates were prepared by pipetting 12 ml of agar into BBL Rodac plates and allowing the agar to solidify. Samples were taken at an indoor cattle facility at five separate locations with a BioScience SAS air-sampling instrument. For each plate, 60 liters of air was sampled. Three replications of the experiment were performed. The TAL method resulted in higher counts of microorganisms on all media tested. In addition, 175 isolates were selected randomly and identified in order to test the selectivity of TAL and the selective media for target organisms. The data obtained in this study show that the TAL resuscitation method is effective and necessary for the recovery of airborne organisms that may be injured.

  16. Evaluation of an automated agar plate streaker.

    PubMed Central

    Tilton, R C; Ryan, R W

    1978-01-01

    An automated agar plate streaker was evaluated. The Autostreaker mechanizes the agar plate streaking process by providing storage for plates, labeling and streaking one or more plates for either isolation or quantitation, and stacking in one of several racks for subsequent incubation. Results showed the Autostreaker to produce agar plates with well-separated colonies and accurate colony counts. A total of 1,930 clinical specimens were processed either in parallel with manual methods or solely by the Autostreaker. Technologist acceptance of machine-streaked plates was outstanding. Images PMID:348722

  17. Synthesis of nano-sized amorphous boron powders through active dilution self-propagating high-temperature synthesis method

    SciTech Connect

    Wang, Jilin; Gu, Yunle; Li, Zili; Wang, Weimin; Fu, Zhengyi

    2013-06-01

    Graphical abstract: Nano-sized amorphous boron powders were synthesized by active dilution self-propagating high-temperature synthesis (SHS) method. The effects of endothermic reaction rate, the possible chemical reaction mechanism and active dilution model for synthesis of the product were also discussed. Highlights: ► Nano-sized amorphous boron powders were synthesized by active dilution self-propagating high-temperature synthesis method. ► The morphology, particle size and purity of the samples could be effectively controlled via changing the endothermic rate. ► The diluter KBH{sub 4} played an important role in active dilution synthesis of amorphous nano-sized boron powders. ► The active dilution method could be further popularized and become a common approach to prepare various inorganic materials. - Abstract: Nano-sized amorphous boron powders were synthesized by active dilution self-propagating high-temperature synthesis (SHS) method at temperatures ranging from 700 °C to 850 °C in a SHS furnace using Mg, B{sub 2}O{sub 3} and KBH{sub 4} as raw materials. Samples were characterized by X-ray powder diffraction (XRD), Laser particle size analyzer, Fourier transform infrared spectra (FTIR), X-ray energy dispersive spectroscopy (EDX), scanning electron microscopy (SEM), transmission electron microscopy (TEM) and high-resolution transmission TEM (HRTEM). The boron powders demonstrated an average particle size of 50 nm with a purity of 95.64 wt.%. The diluter KBH{sub 4} played an important role in the active dilution synthesis of amorphous nano-sized boron powders. The effects of endothermic reaction rate, the possible chemical reaction mechanism and active dilution model for synthesis of the product were also discussed.

  18. A new tracer technique for monitoring groundwater fluxes: the Finite Volume Point Dilution Method.

    PubMed

    Brouyère, Serge; Batlle-Aguilar, Jordi; Goderniaux, Pascal; Dassargues, Alain

    2008-01-28

    Quantification of pollutant mass fluxes is essential for assessing the impact of contaminated sites on their surrounding environment, particularly on adjacent surface water bodies. In this context, it is essential to quantify but also to be able to monitor the variations with time of Darcy fluxes in relation with changes in hydrogeological conditions and groundwater - surface water interactions. A new tracer technique is proposed that generalizes the single-well point dilution method to the case of finite volumes of tracer fluid and water flush. It is called the Finite Volume Point Dilution Method (FVPDM). It is based on an analytical solution derived from a mathematical model proposed recently to accurately model tracer injection into a well. Using a non-dimensional formulation of the analytical solution, a sensitivity analysis is performed on the concentration evolution in the injection well, according to tracer injection conditions and well-aquifer interactions. Based on this analysis, optimised field techniques and interpretation methods are proposed. The new tracer technique is easier to implement in the field than the classical point dilution method while it further allows monitoring temporal changes of the magnitude of estimated Darcy fluxes, which is not the case for the former technique. The new technique was applied to two experimental sites with contrasting objectives, geological and hydrogeological conditions, and field equipment facilities. In both cases, field tracer concentrations monitored in the injection wells were used to fit the calculated modelled concentrations by adjusting the apparent Darcy flux crossing the well screens. Modelling results are very satisfactory and indicate that the methodology is efficient and accurate, with a wide range of potential applications in different environments and experimental conditions, including the monitoring with time of changes in Darcy fluxes.

  19. A new tracer technique for monitoring groundwater fluxes: the Finite Volume Point Dilution Method.

    PubMed

    Brouyère, Serge; Batlle-Aguilar, Jordi; Goderniaux, Pascal; Dassargues, Alain

    2008-01-28

    Quantification of pollutant mass fluxes is essential for assessing the impact of contaminated sites on their surrounding environment, particularly on adjacent surface water bodies. In this context, it is essential to quantify but also to be able to monitor the variations with time of Darcy fluxes in relation with changes in hydrogeological conditions and groundwater - surface water interactions. A new tracer technique is proposed that generalizes the single-well point dilution method to the case of finite volumes of tracer fluid and water flush. It is called the Finite Volume Point Dilution Method (FVPDM). It is based on an analytical solution derived from a mathematical model proposed recently to accurately model tracer injection into a well. Using a non-dimensional formulation of the analytical solution, a sensitivity analysis is performed on the concentration evolution in the injection well, according to tracer injection conditions and well-aquifer interactions. Based on this analysis, optimised field techniques and interpretation methods are proposed. The new tracer technique is easier to implement in the field than the classical point dilution method while it further allows monitoring temporal changes of the magnitude of estimated Darcy fluxes, which is not the case for the former technique. The new technique was applied to two experimental sites with contrasting objectives, geological and hydrogeological conditions, and field equipment facilities. In both cases, field tracer concentrations monitored in the injection wells were used to fit the calculated modelled concentrations by adjusting the apparent Darcy flux crossing the well screens. Modelling results are very satisfactory and indicate that the methodology is efficient and accurate, with a wide range of potential applications in different environments and experimental conditions, including the monitoring with time of changes in Darcy fluxes. PMID:17949849

  20. Evaluation of blood agar microtiter plates for culturing leishmania parasites to titrate parasite burden in spleen and peripheral blood of patients with visceral leishmaniasis.

    PubMed

    Maurya, Radheshyam; Mehrotra, Sanjana; Prajapati, Vijay Kumar; Nylén, Susanne; Sacks, David; Sundar, Shyam

    2010-05-01

    Serial dilution of blood and spleen biopsy specimens, plated on Novy-MacNeal-Nicolle (NNN) blood agar using microtiter culture plates, is a sensitive and reproducible method for detection and growth of Leishmania parasites. Plates could be easily monitored, and growth could be rapidly detected. Moreover, parasite number may be estimated using this technique.

  1. End-point dilution and plaque assay methods for titration of cricket paralysis virus in cultured Drosophila cells.

    PubMed

    Scotti, P D

    1977-05-01

    Cricket paralysis virus, an insect picorna-like virus, caused a distinct c.p.e. in cultured Drosophila melanogaster cells, allowing the development of titration methods based on end-point dilution or plaque assay methods. The end-point dilution (TCD50) method is more sensitive and economical than plaque assays and is easily scored. The data indicate a minimum infectivity/particle ratio of about 1/2000.

  2. Syneresis in agar hydrogels.

    PubMed

    Boral, Shilpi; Saxena, Anita; Bohidar, H B

    2010-03-01

    Agar hydrogels exhibit syneresis which creates internal osmotic stress on the physical network. It was observed that such a stress gives rise to characteristic pulsating modes (breathing modes). Experiments carried over a period of 60-day revealed that the network deformations grew monotonously when the solvent released by syneresis was removed periodically from gel surface. However, when the solvent was not withdrawn, the gel exhibited very slowly relaxing breathing modes. The swelling-deswelling dynamics has been discussed in the generalized framework of a dissipative damped oscillator.

  3. An isotope-dilution standard GC/MS/MS method for steroid hormones in water

    USGS Publications Warehouse

    Foreman, William T.; Gray, James L.; ReVello, Rhiannon C.; Lindley, Chris E.; Losche, Scott A.

    2013-01-01

    An isotope-dilution quantification method was developed for 20 natural and synthetic steroid hormones and additional compounds in filtered and unfiltered water. Deuterium- or carbon-13-labeled isotope-dilution standards (IDSs) are added to the water sample, which is passed through an octadecylsilyl solid-phase extraction (SPE) disk. Following extract cleanup using Florisil SPE, method compounds are converted to trimethylsilyl derivatives and analyzed by gas chromatography with tandem mass spectrometry. Validation matrices included reagent water, wastewater-affected surface water, and primary (no biological treatment) and secondary wastewater effluent. Overall method recovery for all analytes in these matrices averaged 100%; with overall relative standard deviation of 28%. Mean recoveries of the 20 individual analytes for spiked reagent-water samples prepared along with field samples analyzed in 2009–2010 ranged from 84–104%, with relative standard deviations of 6–36%. Detection levels estimated using ASTM International’s D6091–07 procedure range from 0.4 to 4 ng/L for 17 analytes. Higher censoring levels of 100 ng/L for bisphenol A and 200 ng/L for cholesterol and 3-beta-coprostanol are used to prevent bias and false positives associated with the presence of these analytes in blanks. Absolute method recoveries of the IDSs provide sample-specific performance information and guide data reporting. Careful selection of labeled compounds for use as IDSs is important because both inexact IDS-analyte matches and deuterium label loss affect an IDS’s ability to emulate analyte performance. Six IDS compounds initially tested and applied in this method exhibited deuterium loss and are not used in the final method.

  4. Discharge measurement with salt dilution method in irrigation canals: direct sampling and geophysical controls

    NASA Astrophysics Data System (ADS)

    Comina, C.; Lasagna, M.; De Luca, D. A.; Sambuelli, L.

    2013-08-01

    An important starting point for designing management improvements, particularly in irrigation areas, is to record the baseline state of the water resources, including the amount of discharge from canals. In this respect discharge measurements by means of the salt dilution method is a traditional and well-documented technique. However, this methodology can be strongly influenced by the natural streaming characteristics of the canal (e.g. laminar vs. turbulent flow) and accurate precautions must be considered in the choice of both the measuring section and the length of the measuring reach of the canal which can affect the plume shape. The knowledge of plume distribution in the measuring cross-section is of primary importance for a correct location of sampling points aimed in obtaining a reliable measurement. To obtain this, geophysical imaging of an NaCl plume from a slug-injection salt dilution test has been performed within this paper by means of cross-flow fast electric resistivity tomography (FERT) in a real case history. Direct sampling of the same plume has been also performed with a multisampling optimization technique to obtain an average value over the measuring section by means of contemporarily sampling water in nine points. Results show that a correct visualization of the passage of the salt plume is possible by means of geophysical controls and that this can potentially help in the correct location of sampling points.

  5. A stable isotope dilution method for measuring bioavailability of organic contaminants

    PubMed Central

    Delgado-Moreno, Laura; Gan, Jay

    2014-01-01

    Methods for determining bioavailability of organic contaminants suffer various operational limitations. We explored the use of stable isotope labeled references in developing an isotope dilution method (IDM) to measure the exchangeable pool (E) of pyrene and bifenthrin as an approximation of their bioavailability in sediments. The exchange of deuterated bifenthrin or pyrene with its native counterpart was completed within 48 h. The derived E was 38–82% for pyrene and 28–59% for bifenthrin. Regression between E and the sum of rapid and slow desorption fractions obtained from sequential desorption showed a slope close to 1.0. The ability of IDM to predict bioavailability was further shown from a strong relationship (r2 > 0.93) between E and bioaccumulation into Chironomus tentans. Given the abundance of stable isotope labeled references and their relatively easy analysis, the IDM has the potential to become a readily adoptable tool for estimating organic contaminants bioaccessibility in various matrices. PMID:23434573

  6. Precision of a field method for determination of pH in dilute lakes

    USGS Publications Warehouse

    Turk, J.T.

    1986-01-01

    Replicate pH measurements in three dilute lakes made during extreme conditions indicate that pH can be measured in the field with a variance due to measurement error of 0.005 unit. Error of the field technique in measuring the pH of dilute solutions in the laboratory ranges from less than 0.01 unit in dilute strong-acid solutions to about 0.05 unit in air-saturated deionized water.

  7. An improved method of determining vapor-liquid equilibria for dilute organics in aqueous solution.

    PubMed

    Kieckbusch, T G; King, C J

    1979-05-01

    Chromatographic quantitative analyses of aqueous samples containing dilute concentrations of organic solutes are frequently confounded by sorption phenomena, particularly for vapor samples and aged syringes. A novel apparatus was developed for overcoming this problem; it utilizes continual circulation of gas through a thermostated syringe for sampling, followed by a needleless injection into the chromatograph. Precision and reproducibility well under 0.5% can be obtained in this way. This method was applied to a determination of the solubility of isopentyl acetate in water (2600 ppm) by following the equilibrium vapor response for increasing liquid concentrations. It was also used to determine equilibrium partition coefficients for C1-C5 n-acetates between air and water, over a range of temperatures from 25 to 40 degrees C. PMID:19847988

  8. Comparison of fluorescence optical respirometry and microbroth dilution methods for testing antimicrobial compounds.

    PubMed

    Hałasa, R; Turecka, K; Orlewska, C; Werel, W

    2014-12-01

    An analysis of the usefulness of the fluorescence optical respirometry test method to study several antimicrobials was performed. An oxygen-sensitive sensor: ruthenium-tris(4,7-diphenyl-1,10-phenanthroline) dichloride (Ru(DPP)3Cl2), the phosphorescence of which is quenched by molecular oxygen, was synthesised according to a method modified by us and then applied. A prototype sensitive measurement system was designed and constructed. Analyses of the impact of various antimicrobial chemical factors were performed: ampicillin, co-trimoxazole, nystatin, and newly synthesised compounds. It was shown that optical respirometry allows for analysis of the culture growth kinetics of bacteria and fungi and determination of cell growth parameters. It was shown also that MIC values obtained by fluorescence optical respirometry are consistent with the results of the MIC determinations made by serial dilution method (traditional MIC testing using CLSI). The method allows the time to obtain results to be significantly reduced (from 24-48 h to 5-7 h for bacteria and 24 yeasts) and allows the effect of concentrations below the MIC for the metabolic activity of microorganisms to be monitored. The sensitivity of the method allowed the volume of the tested samples to be lessened from 160 μl to 50 μl. Fluorescence optical respirometry allows for the rapid detection and evaluation of the action of various chemical compounds on the metabolic activity of microorganisms in real-time measurement of fluorescence intensity.

  9. Development of large area nanostructured silicon-hydrogen alloy material with improved stability for solar cell application by argon dilution method

    NASA Astrophysics Data System (ADS)

    Dey, Arka; Das, Mrinmay; Datta, Joydeep; Jana, Rajkumar; Dhar, Joydeep; Sil, Sayantan; Biswas, Debasish; Banerjee, Chandan; Ray, Partha Pratim

    2016-07-01

    Here we have presented the results of large area (30 × 30 cm2) silicon-hydrogen alloy material and solar cell by argon dilution method. As an alternative to hydrogen dilution, argon dilution method has been applied to develop single junction solar cell with appreciable stability. Optimization of deposition conditions revealed that 95% argon dilution gives a nanostructured material with improved transport property and less light induced degradation. The minority carrier diffusion length (L d ) and mobility-lifetime (μτ) product of the material with 95% argon dilution degrades least after light soaking. Also the density of states (DOS) below conduction level reveals that this material is less defective. Solar cell with this argon diluted material has been fabricated with all the layers deposited by argon dilution method. Finally we have compared the argon diluted solar cell results with the optimized hydrogen diluted solar cell. Light soaking study proves that it is possible to develop stable solar cell on large area by argon dilution method and that the degradation of argon diluted solar cell is less than that of hydrogen diluted one. [Figure not available: see fulltext.

  10. 21 CFR 184.1115 - Agar-agar.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Agar-agar. 184.1115 Section 184.1115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of...

  11. 21 CFR 184.1115 - Agar-agar.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Agar-agar. 184.1115 Section 184.1115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of...

  12. 21 CFR 184.1115 - Agar-agar.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Agar-agar. 184.1115 Section 184.1115 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DIRECT FOOD... ingredient meets the specifications of the “Food Chemicals Codex,” 3d Ed. (1981), p. 11, which...

  13. New method of tracing blood hemoglobin concentration to hematocrit ratio for monitoring plasma dilution and osmotic origin shifts in blood.

    PubMed

    Andrijauskas, Audrius; Ivaskevicius, Juozas

    2006-01-01

    Blood hemoglobin concentration and hematocrit are probably the most widely used parameters for outpatient and inpatient examination. In addition to their inherent significance for evaluation of blood viscosity and oxygen carrying capacity, these parameters are traditionally used as tracers of plasma dilution. Blood test derived results are conventionally recorded on multiple pages in patient's medical records making dynamical investigations tedious and time-consuming. In addition, research results describing plasma dilution by means of hemoglobin or hematocrit are presented in a clinically unpractical way. A new method, referred to as HBS Graphics (patent pending--USA serial # 60/712809) is introduced for the first time in this article. This method of evaluation of dynamical hemoglobin concentration, hematocrit and mean corpuscular hemoglobin concentration value deploys interfering parameter shifts for the evaluation of plasma dilution in relation to osmotic dynamics. The HBS Graphics complements two coordinate systems--hemoglobin concentration and hematocrit--with incorporated mean corpuscular hemoglobin concentration value specific trends referred to as radiating lines. Isosmotic plasma dilution and erythrocyte volume shifts follow radiating lines, while osmotic shifts induce intertrend shifts. This article also reviews other methods of tracing plasma dilution by means of blood hemoglobin concentration and hematocrit dynamics.

  14. A novel method for the quantification of quinic acid in food using stable isotope dilution analysis.

    PubMed

    Erk, Thomas; Bergmann, Hannah; Richling, Elke

    2009-01-01

    Organic acids play an important role in the flavor and taste of plant-derived foods. Quinic acid (QA) is one of the major acids. In the past, several methods like HPLC/UV, GC, and capillary electrophoresis were used for identification and quantification of QA. For the first time, a novel, sensitive, and selective method for the quantification of QA in food using stable isotope dilution analysis with HPLC/MS/MS has been established. Uniformly labeled 13C-QA was used as a standard to reduce sample preparations and to overcome matrix and ionization effects. The method was used to determine the QA content of red wines, instant coffees, and cloudy apple juices. QA contents of instant coffees were 64.4 and 63.6 g/kg powder. The concentrations in red wines were 24.0 and 25.1 mg/L, and 1493.3 and 1705.2 mg/L in cloudy apple juices.

  15. Enumeration of sublethally injured Escherichia coli O157:H7 ATCC 43895 and Escherichia coli strain B-41560 using selective agar overlays versus commercial methods.

    PubMed

    Smith, Amanda R; Ellison, Alysha L; Robinson, Amanda L; Drake, Maryanne; McDowell, Susan A; Mitchell, James K; Gerard, Patrick D; Heckler, Rachel A; McKillip, John L

    2013-04-01

    Quality control procedures during food processing may involve direct inoculation of food samples onto appropriate selective media for subsequent enumeration. However, sublethally injured bacteria often fail to grow, enabling them to evade detection and intervention measures and ultimately threaten the health of consumers. This study compares traditional selective and nonselective agar-based overlays versus two commercial systems (Petrifilm and Easygel) for recovery of injured E. coli B-41560 and O157:H7 strains. Bacteria were propagated in tryptic soy broth (TSB), ground beef slurry, and infant milk formula to a density of 10(6) to 10(8) CFU/ml and then were stressed for 6 min either in lactic acid (pH 4.5) or heat shocked for 3 min at 60°C. Samples were pour plated in basal layers of either tryptic soy agar (TSA), sorbitol MacConkey agar (SMAC), or violet red bile agar (VRB) and were resuscitated for 4 h prior to addition of agar overlays. Other stressed bacteria were plated directly onto Petrifilm and Easygel. Results indicate that selective and nonselective agar overlays recovered significantly higher numbers (greater than 1 log) of acid- and heat-injured E. coli O157:H7 from TSB, ground beef, and infant milk formula compared with direct plating onto selective media, Petrifilm, or Easygel, while no significant differences among these media combinations were observed for stressed E. coli B-41560. Nonstressed bacteria from TSB and ground beef were also recovered at densities significantly higher in nonselective TSA-TSA and in VRB-VRB and SMAC-SMAC compared with Petrifilm and Easygel. These data underscore the need to implement food safety measures that address sublethally injured pathogens such as E. coli O157:H7 in order to avoid underestimation of true densities for target pathogens.

  16. Effect of refrigerated storage on cefaclor in Mueller-Hinton agar.

    PubMed Central

    Surprenant, A M; Preston, D A

    1985-01-01

    Cefaclor is less stable than most cephalosporins in media at 35 degrees C. We demonstrated that the activity of cefaclor in Mueller-Hinton agar diminishes continuously at 4 degrees C, resulting in a loss of two-thirds of the activity within 21 days. We recommend that agar dilution plates for this cephalosporin be prepared on the day of their use. PMID:3968200

  17. Pantothenic acid quantification: method comparison of a stable isotope dilution assay and a microbiological assay.

    PubMed

    Rychlik, Michael; Roth-Maier, Dora

    2005-05-01

    Different foods and feedstuffs were analyzed for pantothenic acid (PA) by the recently developed stable isotope dilution assay (SIDA) and by the standard method, a microbiological assay (MA). The SIDA involved the use of [13C3, 15N]-pantothenic acid as the internal standard and detection by liquid chromatography-tandem mass spectrometry. The analysis of identical extracts minimized systematic bias due to equal extraction yields and enabled an ideal comparison between both methods. For the samples derived from plants a good accordance between the MA and the SIDA of total PA was found, whereas for the products of animal origin, higher contents were measured by MA than by SIDA. From the results of treatments by pantetheinase and phosphatase on the one hand and papain and diastase on the other, it was concluded that MA is able to measure a significant amount of bound PA. Furthermore, the data imply that microbial enzymes were able to cleave PA conjugates more effectively than pantetheinase and phosphatase treatment. PMID:16028638

  18. A comparison of a new centrifuge sugar flotation technique with the agar method for the extraction of immature Culicoides (Diptera: Ceratopogonidae) life stages from salt marsh soils.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two sampling techniques, agar extraction (AE) and centrifuge sugar flotation extraction (CSFE) were compared to determine their relative efficacy to recover immature stages of Culicoides spp from salt marsh substrates. Three types of samples (seeded with known numbers of larvae, homogenized field s...

  19. Quantification of methane and nitrous oxide emissions from various waste treatment facilities by tracer dilution method

    NASA Astrophysics Data System (ADS)

    Mønster, Jacob; Rella, Chris; Jacobson, Gloria; Kjeldsen, Peter; Scheutz, Charlotte

    2013-04-01

    Urban activities generate solid and liquid waste, and the handling and aftercare of the waste results in the emission of various compounds into the surrounding environment. Some of these compounds are emitted as gasses into the atmosphere, including methane and nitrous oxide. Methane and nitrous oxide are strong greenhouse gases and are considered to have 25 and 298 times the greenhouse gas potential of carbon dioxide on a hundred years term (Solomon et al. 2007). Global observations of both gasses have shown increasing concentrations that significantly contribute to the greenhouse gas effect. Methane and nitrous oxide are emitted from both natural and anthropogenic sources and inventories of source specific fugitive emissions from the anthropogenic sources of methane and nitrous oxide of are often estimated on the basis of modeling and mass balance. Though these methods are well-developed, actual measurements for quantification of the emissions is a very useful tool for verifying the modeling and mass balance as well as for validation initiatives done for lowering the emissions of methane and nitrous oxide. One approach to performing such measurements is the tracer dilution method (Galle et al. 2001, Scheutz et al. 2011), where the exact location of the source is located and a tracer gas is released at this source location at a known flow. The ratio of downwind concentrations of the tracer gas and the methane and nitrous oxide gives the emissions rates of the greenhouse gases. This tracer dilution method can be performed using both stationary and mobile measurements and in both cases, real-time measurements of both tracer and quantified gas are required, placing high demands on the analytical detection method. To perform the methane and nitrous oxide measurements, two robust instruments capable of real-time measurements were used, based on cavity ring-down spectroscopy and operating in the near-infrared spectral region. One instrument measured the methane and

  20. An evaluation of blood smears made by a new method using a spinner and diluted blood.

    PubMed

    Nourbakhsh, M; Atwood, J G; Raccio, J; Seligson, D

    1978-12-01

    Blood smears were prepared with the use of a spinner, which rotated with a fixed velocity for a fixed time. All blood samples used for spun smears were diluted with a fixed ratio of buffered isotonic saline solution. Distribution of cells in these smears was found to be random. The average number of cells per unit area was substantially uniform from place to place on the same slide and on multiple slides made with the smae sample. The distribution of leukocytes by type was also iniform. For different blood samples, the average number of cells per unit area in the smears correlated well with the measured cell concentrations per unit volume in the samples for leukocytes, erythrocytes and platelets. Leukocyte differential counts on replicate spun smears using the same bloods also agreed to within the sampling error. They similarly agreed with differential counts on pulled smears made from undiluted samples of the same bloods. With few exceptions, erythrocytic morphology on the spun smears was comparable to that on the good areas of pulled smears made with undiluted samples of the same bloods. Nearly all the spun smears were suitable for both viual and fully automated hematologic examination for leukocytes, erythrocytes, and platelets. This was true over nearly the whole area of each spun slide. In these ways this spinner method makes smears whose consistently high quality is little affected by either the properties of the blood sample or the skill of maker.

  1. Shear viscosity of dilute suspensions of ellipsoidal particles with a lattice Boltzmann method.

    PubMed

    Huang, Haibo; Wu, YanFeng; Lu, Xiyun

    2012-10-01

    The intrinsic viscosities for prolate and oblate spheroidal suspensions in a dilute Newtonian fluid are studied using a three-dimensional lattice Boltzmann method. Through directly calculated viscous dissipation, the minimum and maximum intrinsic viscosities and the period of the tumbling state all agree well with the analytical solution for particles with different aspect ratios. This numerical test verifies the analysis on maximum and minimum intrinsic viscosities. Different behavior patterns of transient intrinsic viscosity in a period are analyzed in detail. A phase lag between the transient intrinsic viscosity and the orientation of the particle at finite Reynolds number (Re) is found and attributed to fluid and particle inertia. At lower Re, the phase lag increases with Re. There exists a critical Reynolds number Rea at which the phase lag begins to decrease with Re. The Rea depends on the aspect ratio of the particle. We found that both the intrinsic viscosity and the period change linearly with Re when ReRea (high-Re regime). In the high-Re regime, the dependence of the period on Re is consistent with a scaling law, and the dependence of the intrinsic viscosity on Re is well described by second-degree polynomial fits. PMID:23214675

  2. Shear viscosity of dilute suspensions of ellipsoidal particles with a lattice Boltzmann method

    NASA Astrophysics Data System (ADS)

    Huang, Haibo; Wu, YanFeng; Lu, Xiyun

    2012-10-01

    The intrinsic viscosities for prolate and oblate spheroidal suspensions in a dilute Newtonian fluid are studied using a three-dimensional lattice Boltzmann method. Through directly calculated viscous dissipation, the minimum and maximum intrinsic viscosities and the period of the tumbling state all agree well with the analytical solution for particles with different aspect ratios. This numerical test verifies the analysis on maximum and minimum intrinsic viscosities. Different behavior patterns of transient intrinsic viscosity in a period are analyzed in detail. A phase lag between the transient intrinsic viscosity and the orientation of the particle at finite Reynolds number (Re) is found and attributed to fluid and particle inertia. At lower Re, the phase lag increases with Re. There exists a critical Reynolds number Rea at which the phase lag begins to decrease with Re. The Rea depends on the aspect ratio of the particle. We found that both the intrinsic viscosity and the period change linearly with Re when ReRea (high-Re regime). In the high-Re regime, the dependence of the period on Re is consistent with a scaling law, and the dependence of the intrinsic viscosity on Re is well described by second-degree polynomial fits.

  3. A new method to determine the yield stress of diluted polymeric solutions

    NASA Astrophysics Data System (ADS)

    Soto, Enrique; Ruiz, Servando; Cordova Aguilar, Maria Soledad

    2012-11-01

    A new method to measure the yield stress for diluted polymeric solutions is presented. The tested solutions exhibit shear thinning behavior a once the critical yield stress is overcame. In rheology, these fluids are known as Herschel-Buckley. The yield stress phenomenon and its relation with bubble motion is an important issue for different industries, for example, personal care, paints and some others. As a result of the yield stress, small bubbles remain trapped in the fluid bulk, but above a critical volume, which is related with the characteristic yield stress, the bubbles flow in the liquid. In order to change the bubble volume, the liquid is placed in a cylindrical container whose pressure is decreased by a vacuum pump. The bubble growths as the pressure decreases and keeps its position until it reaches the critical volume. The bubble shape changes with volume and velocity, and a competition among surface, gravitational, inertial and viscous forces is discussed. The yield stress determined value is higher than the obtained from simple shear measurements due to the complex flow around the bubble.

  4. [Titration of Ebola and Marburg viruses by plaque formation under semi liquid agar].

    PubMed

    Ustinova, E N; Shestopalov, A M; Bakulina, L F; Chepurnov, A A

    2003-01-01

    The method of titration of Ebola and Marburg viruses using plaque formation under semifluid agar cover is considered. Advantages of this method over conventional method of titration of these viruses with the use of hard agar cover are discussed.

  5. Susceptibility of a polycaprolactone-based root canal filling material to degradation using an agar-well diffusion assay

    PubMed Central

    Hiraishi, Noriko; Sadek, Fernanda T.; King, Nigel M.; Ferrari, Marco; Pashley, David H.; Tay, Franklin R

    2013-01-01

    Purpose Cholesterol esterase is both a component of salivary hydrolases as well as an inflammatory cell-derived enzyme and has been shown to cause biodegradation of methacrylate-based resin composites. This study examined whether Resilon, a polycaprolactone-based thermoplastic root filling material is susceptible to biodegradation by cholesterol esterase using agar-well diffusion assay of serially-diluted aqueous Resilon emulsions that were dispersed in agar. Materials and methods Emulsions of Resilon and polycaprolactone were prepared and dispersed in agar on culture plates. Two different concentrations of a cholesterol esterase (0.3 and 1.2 U/mL) were prepared and fed to wells prepared in the agar plates using an agar-well diffusion assay for examination the degradation of polymeric materials. Results Degradation of the emulsified Resilon was manifested as the formation of clear zones of different sizes around the agar wells. No clear zones were observed in agar wells that contain sterile distilled water as the negative control. Clinical significance Although dispersion Resilon into an emulsion is not the way in which this material is employed as a root filling material, the potential for Resilon to be degraded by cholesterol esterase is of potential concern as one cannot limit the degradation of extruded Resilon from a root apex by monocyte-derived macrophages to just the anatomical root apex. As the present study employed a high concentration of cholesterol esterase, further studies should be directed to examining the degradation of Resilon using macrophage cell cultures. PMID:18578181

  6. Using the tracer-dilution discharge method to develop streamflow records for ice-affected streams in Colorado

    USGS Publications Warehouse

    Capesius, Joseph P.; Sullivan, Joseph R.; O'Neill, Gregory B.; Williams, Cory A.

    2005-01-01

    Accurate ice-affected streamflow records are difficult to obtain for several reasons, which makes the management of instream-flow water rights in the wintertime a challenging endeavor. This report documents a method to improve ice-affected streamflow records for two gaging stations in Colorado. In January and February 2002, the U.S. Geological Survey, in cooperation with the Colorado Water Conservation Board, conducted an experiment using a sodium chloride tracer to measure streamflow under ice cover by the tracer-dilution discharge method. The purpose of this study was to determine the feasibility of obtaining accurate ice-affected streamflow records by using a sodium chloride tracer that was injected into the stream. The tracer was injected at two gaging stations once per day for approximately 20 minutes for 25 days. Multiple-parameter water-quality sensors at the two gaging stations monitored background and peak chloride concentrations. These data were used to determine discharge at each site. A comparison of the current-meter streamflow record to the tracer-dilution streamflow record shows different levels of accuracy and precision of the tracer-dilution streamflow record at the two sites. At the lower elevation and warmer site, Brandon Ditch near Whitewater, the tracer-dilution method overestimated flow by an average of 14 percent, but this average is strongly biased by outliers. At the higher elevation and colder site, Keystone Gulch near Dillon, the tracer-dilution method experienced problems with the tracer solution partially freezing in the injection line. The partial freezing of the tracer contributed to the tracer-dilution method underestimating flow by 52 percent at Keystone Gulch. In addition, a tracer-pump-reliability test was conducted to test how accurately the tracer pumps can discharge the tracer solution in conditions similar to those used at the gaging stations. Although the pumps were reliable and consistent throughout the 25-day study period

  7. [Detection of TDH-producing Vibrio parahaemolyticus O3:K6 from naturally contaminated shellfish using an immunomagnetic separation method and chromogenic agar medium].

    PubMed

    Hara-Kudo, Y; Sugiyama, K; Nishina, T; Saitoh, A; Nakagawa, H; Ichihara, T; Konuma, H; Hasegawa, J; Kumagai, S

    2001-11-01

    We attempted to isolate TDH-producing Vibrio parahaemolyticus O3:K6 from shellfish. Asari samples were incubated with TSB supplemented with 2% (w/v) NaCl for 6 h, and then the 6-h cultures were incubated with salt polymyxin broth for 18 h. After the two-step enrichment, a 1 ml portion of the culture was treated with magnetic beads coated with K6 antibody for immunoconcentration of V. parahaemolyticus O3:K6. The immunoconcentrated and untreated cultures were plated onto a chromogenic agar and TCBS agar media for isolation of V. parahaemolyticus. TDH-producing V. parahaemolyticus O3:K6 was isolated from 3 out of 66 lots (4.5%) of naturally contaminated Asari. Six of 4,265 colonies suspected as V. parahaemolyticus (0.14%) were TDH-producing V. parahaemolyticus O3:K6.

  8. Abilities of the mCP Agar method and CRENAME alpha toxin-specific real-time PCR assay to detect Clostridium perfringens spores in drinking water.

    PubMed

    Maheux, Andrée F; Bérubé, Eve; Boudreau, Dominique K; Villéger, Romain; Cantin, Philippe; Boissinot, Maurice; Bissonnette, Luc; Bergeron, Michel G

    2013-12-01

    We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a Clostridium perfringens-specific real-time PCR (rtPCR) assay based on the cpa gene (cpa rtPCR) by using a bacterial strain panel composed of C. perfringens and non-C. perfringens Clostridium strains. All non-C. perfringens Clostridium strains tested negative, whereas all C. perfringens strains tested positive with the cpa rtPCR, for an analytical specificity and ubiquity of 100%. The cpa rtPCR assay was then used to confirm the identity of 116 putative C. perfringens isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Four mCP(-)/rtPCR(+) colonies were identified as C. perfringens, whereas 3 mCP(+)/rtPCR(-) colonies were identified as non-C. perfringens. The cpa rtPCR was negative with all 51 non-C. perfringens strains and positive with 64 of 65 C. perfringens strains. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME + cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water. CRENAME + cpa rtPCR detected as few as one C. perfringens CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME + cpa rtPCR also allows the simultaneous and sensitive detection of Escherichia coli and C. perfringens from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection.

  9. Antibiotic activity of tigecycline against clinical pathogens by the micro dilution method.

    PubMed

    Mantzourani, I; Plessas, S; Alexopoulos, A; Voidarou, C; Bezirtzoglou, E

    2011-12-01

    Resistant pathogens are the cause of clinical infections which threatening the patients lives and challenging the health systems through their economic importance. Therefore, new antibacterial agents with a broader spectrum of activity that protect against development of resistance are required. Tigecycline (Tygacil, Wyeth) is a relatively new FDA and EMEA approved glycylcycline antimicrobial with an expanded broad-spectrum activity against pathogens involved in complicated skin and skin structure infections. In this study we evaluated the in vitro activity of tigecycline in comparison to 14 other antibiotics against 182 clinical pathogens by use of the micro dilution method. In overall, tigecycline exhibited the lowest Minimum Inhibitory Concentration (MIC) values in almost all bacteria with a mean of 0.52 ± 1.25mg/L, followed by meropenem and levofloxacin (mean MIC values 1.29 ± 2.52 and 1.45 ± 3.078 mg/L, respectively). MIC50 and MIC90 values of tigecycline were: 0.06 and 0.15 mg/L for E. coli, 0.12 and 1.00 mg/L for Klebsiella sp., 0.12 and 0.85 mg/L for various Enterobacter sp., 1.00 and 8.00 mg/L for Pseudomonas sp., 0.25 and 1.00 mg/L for Acinetobacter sp., 0.06 and 0.12 mg/L for Serratia sp., 0.12 and 0.25mg/L for Staphylococcus aureus, 0.5 and 5.00 mg/L for Streptococcus sp. The MIC values recorded were among the lowest in recent literature for Acinetobacter sp. (included A. baumannii), and comparable to those obtained for Klebsiella, Serratia and Enterobacter indicating that tigecycline has a promising in vitro activity. PMID:21513809

  10. Geophysical methods to support correct water sampling locations for salt dilution gauging

    NASA Astrophysics Data System (ADS)

    Comina, C.; Lasagna, M.; De Luca, D. A.; Sambuelli, L.

    2014-05-01

    To improve water management design, particularly in irrigation areas, it is important to evaluate the baseline state of the water resources, including canal discharge. Discharge measurements, using salt dilution gauging, are a traditional and well-documented technique. The complete mixing of salt used for dilution gauging is required for reliable measurements; this condition is difficult to test or verify and, if not fulfilled, is the largest source of uncertainty in the discharge calculation. In this paper, a geophysical technique (FERT, Fast Electrical Resistivity Tomography) is proposed for imaging the distribution of the salt plume used for dilution gauging at every point along a sampling cross-section. In this way, it is possible to check whether complete mixing has occurred. If the mixing is not complete, the image created by FERT can also provide guidance for selecting water-sampling locations in the sampling cross-section. A water multi-sampling system prototype for the simultaneous sampling of canal water at different points within the cross-section, aimed to potentially take into account concentration variability, is also proposed and tested. Preliminary results of a single test with salt dilution gauging and FERT in a real case are reported. The results show that imaging the passage of the salt plume is possible by means of geophysical controls and that this can potentially help in the selection of water sampling points.

  11. MAXIMUM INHIBITORY DILUTION OF MOUTHWASHES CONTAINING CHLORHEXIDINE AND POLYHEXAMETHYLENE BIGUANIDE AGAINST SALIVARY STAPHYLOCOCCUS AUREUS

    PubMed Central

    Nascimento, Andresa Piacezzi; Tanomaru, Juliane Maria Guerreiro; Matoba, Fumio; Watanabe, Evandro; Tanomaru, Mario; Ito, Izabel Yoko

    2008-01-01

    Objective: The aim of the present study was to determine the in vitro maximum inhibitory dilution (MID) of two chlorhexidine-based oral mouthwashes (CHX): Noplak®, Periogard®, and one polyhexamethylene biguanide-based mouthwash (PHMB): Sanifill Premium® against 28 field Staphylococcus aureus strains using the agar dilution method. Materials and Methods: For each product, decimal dilutions ranging from 1/10 to 1/655,360 were prepared in distilled water and added to Mueller Hinton Agar culture medium. After homogenization, the culture medium was poured onto Petri dishes. Strains were inoculated using a Steers multipoint inoculator and dishes were incubated at 37°C for 24hours. For reading, MID was considered as the maximum dilution of the mouthwash still capable of inhibiting microbial growth. Results: Sanifill Premium® inhibited the growth of all strains at 1/40 dilution and of 1 strain at 1/80 dilution. Noplak® inhibited the growth of 23 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Periogard® showed inhibited growth of 7 strains at 1/640 dilution and of all 28 strains at 1/320 dilution. Data were submitted to Kruskal-Wallis statistical test, showing significant differences between the mouthwashes evaluated (p<0.05). No significant difference was found between Noplak® and Periogard® (p>0.05). Sanifill Premium® was the least effective (p<0.05). Conclusion: It was concluded that CHX-based mouthwashes present better antimicrobial activity against S. Aureus than the PHMB-based mouthwash. PMID:19089230

  12. Multiregional evaluation of the SimPlate heterotrophic plate count method compared to the standard plate count agar pour plate method in water.

    PubMed

    Jackson, R W; Osborne, K; Barnes, G; Jolliff, C; Zamani, D; Roll, B; Stillings, A; Herzog, D; Cannon, S; Loveland, S

    2000-01-01

    A new SimPlate heterotrophic plate count (HPC) method (IDEXX Laboratories, Westbrook, Maine) was compared with the pour plate method at 35 degrees C for 48 h. Six laboratories tested a total of 632 water samples. The SimPlate HPC method was found to be equivalent to the pour plate method by regression analysis (r = 0. 95; y = 0.99X + 0.06).

  13. Abilities of the mCP Agar Method and CRENAME Alpha Toxin-Specific Real-Time PCR Assay To Detect Clostridium perfringens Spores in Drinking Water

    PubMed Central

    Maheux, Andrée F.; Bérubé, Ève; Boudreau, Dominique K.; Villéger, Romain; Cantin, Philippe; Boissinot, Maurice; Bissonnette, Luc

    2013-01-01

    We first determined the analytical specificity and ubiquity (i.e., the ability to detect all or most strains) of a Clostridium perfringens-specific real-time PCR (rtPCR) assay based on the cpa gene (cpa rtPCR) by using a bacterial strain panel composed of C. perfringens and non-C. perfringens Clostridium strains. All non-C. perfringens Clostridium strains tested negative, whereas all C. perfringens strains tested positive with the cpa rtPCR, for an analytical specificity and ubiquity of 100%. The cpa rtPCR assay was then used to confirm the identity of 116 putative C. perfringens isolates recovered after filtration of water samples and culture on mCP agar. Colonies presenting discordant results between the phenotype on mCP agar and cpa rtPCR were identified by sequencing the 16S rRNA and cpa genes. Four mCP−/rtPCR+ colonies were identified as C. perfringens, whereas 3 mCP+/rtPCR− colonies were identified as non-C. perfringens. The cpa rtPCR was negative with all 51 non-C. perfringens strains and positive with 64 of 65 C. perfringens strains. Finally, we compared mCP agar and a CRENAME (concentration and recovery of microbial particles, extraction of nucleic acids, and molecular enrichment) procedure plus cpa rtPCR (CRENAME + cpa rtPCR) for their abilities to detect C. perfringens spores in drinking water. CRENAME + cpa rtPCR detected as few as one C. perfringens CFU per 100 ml of drinking water sample in less than 5 h, whereas mCP agar took at least 25 h to deliver results. CRENAME + cpa rtPCR also allows the simultaneous and sensitive detection of Escherichia coli and C. perfringens from the same potable water sample. In itself, it could be used to assess the public health risk posed by drinking water potentially contaminated with pathogens more resistant to disinfection. PMID:24077714

  14. Development of a selective agar plate for the detection of Campylobacter spp. in fresh produce.

    PubMed

    Yoo, Jin-Hee; Choi, Na-Young; Bae, Young-Min; Lee, Jung-Su; Lee, Sun-Young

    2014-10-17

    This study was conducted to develop a selective medium for the detection of Campylobacter spp. in fresh produce. Campylobacter spp. (n=4), non-Campylobacter (showing positive results on Campylobacter selective agar) strains (n=49) isolated from fresh produce, indicator bacteria (n=13), and spoilage bacteria isolated from fresh produce (n=15) were plated on four Campylobacter selective media. Bolton agar and modified charcoal cefoperazone deoxycholate agar (mCCDA) exhibited higher sensitivity for Campylobacter spp. than did Preston agar and Hunt agar, although certain non-Campylobacter strains isolated from fresh produce by using a selective agar isolation method, were still able to grow on Bolton agar and mCCDA. To inhibit the growth of non-Campylobacter strains, Bolton agar and mCCDA were supplemented with 5 antibiotics (rifampicin, polymyxin B, sodium metabisulfite, sodium pyruvate, ferrous sulfate) and the growth of Campylobacter spp. (n=7) and non-Campylobacter strains (n=44) was evaluated. Although Bolton agar supplemented with rifampicin (BR agar) exhibited a higher selectivity for Campylobacter spp. than did mCCDA supplemented with antibiotics, certain non-Campylobacter strains were still able to grow on BR agar (18.8%). When BR agar with various concentrations of sulfamethoxazole-trimethoprim were tested with Campylobacter spp. (n=8) and non-Campylobacter (n=7), sulfamethoxazole-trimethoprim was inhibitory against 3 of 7 non-Campylobacter strains. Finally, we validated the use of BR agar containing 50mg/L sulfamethoxazole (BRS agar) or 0.5mg/L ciprofloxacin (BRCS agar) and other selective agars for the detection of Campylobacter spp. in chicken and fresh produce. All chicken samples were positive for Campylobacter spp. when tested on mCCDA, BR agar, and BRS agar. In fresh produce samples, BRS agar exhibited the highest selectivity for Campylobacter spp., demonstrating its suitability for the detection of Campylobacter spp. in fresh produce.

  15. Geophysical methods to support correct water sampling locations for salt dilution gauging

    NASA Astrophysics Data System (ADS)

    Comina, C.; Lasagna, M.; De Luca, D. A.; Sambuelli, L.

    2014-08-01

    To improve water management design, particularly in irrigation areas, it is important to evaluate the baseline state of the water resources, including canal discharge. Salt dilution gauging is a traditional and well-documented technique in this respect. The complete mixing of salt used for dilution gauging is required; this condition is difficult to test or verify and, if not fulfilled, is the largest source of uncertainty in the discharge calculation. In this paper, a geophysical technique (FERT, fast electrical resistivity tomography) is proposed for imaging the distribution of the salt plume used for dilution gauging at every point along a sampling cross section. With this imaging, complete mixing can be verified. If the mixing is not complete, the image created by FERT can also provide a possible guidance for selecting water-sampling locations in the sampling cross section. A water multi-sampling system prototype aimed to potentially take into account concentration variability is also proposed and tested. The results reported in the paper show that FERT provides a three-dimensional image of the dissolved salt plume and that this can potentially help in the selection of water sampling points.

  16. Effect of dilution on compressibility of naproxen in acetonitrile studied by ultrasonic method

    NASA Astrophysics Data System (ADS)

    Marczak, W.; Kowalska, T.; Bucek, M.; Piotrowski, D.; Sajewicz, M.

    2006-11-01

    Naproxen, ibuprofen, and ketoprofen are non-steroidal anti-inflammatory drugs. All of them belong to chiral 2-arylpropionic acids (2-APAs). Chiral compounds may remain in a patient's body as two antimers, even if administered as a single one, due to transenantiomerization. That is dangerous if therapeutic enantiomer has a toxic antipode. Chromatographic data suggest that solutions of S-(+)-naproxen in acetonitrile are stiffer than the pure solvent that favours oscillatory transenantiomerisation. Acoustic and volumetric studies of dilute solutions of naproxen in acetonitrile have been undertaken to verify that supposition. The molar adiabatic compressibility and volume depend linearly on the molar percent of naproxen at temperatures from 298.15 K to 313.15 K. Limiting partial compressibility of naproxen is close to zero and decreases slightly with increasing temperature. Thus, the compressibility of dilute solutions is mainly due to compressibility of acetonitrile, while naproxen is virtually incompressible. The hydrogen-bonded dimers of naproxen probably remain intact, even at infinite dilution.

  17. [Effect of ligand concentration on the precision of determining the parameters of ligand-receptor interaction by serial dilution methods].

    PubMed

    Bobrovnik, S A

    2004-01-01

    Earlier we suggested the method of serial dilution, which allows one to determine the parameters of ligand-receptor interaction even if the reactants are in a mixture and their concentrations are unknown. The method is especially useful if the liability of studied receptor does not allow its separation from corresponding ligand. The important prerequisite of the method's precision is that the concentration of the ligand should be sufficiently high comparing to the concentration of the receptor. In the present paper it was demonstrated that the method allows one to obtain sufficiently good precision even in the case when the concentration of the ligand is only one tenth of the receptor concentration.

  18. 48 CFR 401.371 - AGAR Advisories.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 48 Federal Acquisition Regulations System 4 2011-10-01 2011-10-01 false AGAR Advisories. 401.371... ACQUISITION REGULATION SYSTEM Agency Acquisition Regulations 401.371 AGAR Advisories. The SPE may issue AGAR Advisories, consistent with the policies of the FAR and the AGAR, for the following purposes: (a)...

  19. 48 CFR 401.371 - AGAR Advisories.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 4 2010-10-01 2010-10-01 false AGAR Advisories. 401.371... ACQUISITION REGULATION SYSTEM Agency Acquisition Regulations 401.371 AGAR Advisories. The SPE may issue AGAR Advisories, consistent with the policies of the FAR and the AGAR, for the following purposes: (a)...

  20. DETERMINATION OF THE MAXIMUM INHIBITORY DILUTION OF CETYLPYRIDINIUM CHLORIDE-BASED MOUTHWASHES AGAINST STAPHYLOCOCCUS AUREUS: AN IN VITRO STUDY

    PubMed Central

    Watanabe, Evandro; Tanomaru, Juliane Maria Guerreiro; Nascimento, Andresa Piacezzi; Matoba, Fumio; Tanomaru, Mario; Ito, Izabel Yoko

    2008-01-01

    The aim of this in vitro study was to determine the maximum inhibitory dilution (MID) of four cetylpyridinium chloride (CPC)-based mouthwashes: CPC+Propolis, CPC+Malva, CPC+Eucaliptol+Juá+Romã+Propolis (Natural Honey®) and CPC (Cepacol®), against 28 Staphylococcus aureus field strains, using the agar dilution method. Decimal dilutions ranging from 1/10 to 1/ 655,360 were prepared and added to Mueller Hinton Agar. Strains were inoculated using Steers multipoint inoculator. The inocula were seeded onto the surface of the culture medium in Petri dishes containing different dilutions of the mouthwashes. The dishes were incubated at 37°C for 24 h. For readings, the MID was considered as the maximum dilution of mouthwash still capable of inhibiting microbial growth. The obtained data showed that CPC+Propolis had antimicrobial activity against 27 strains at 1/320 dilution and against all 28 strains at 1/160 dilution, CPC+Malva inhibited the growth of all 28 strains at 1/320 dilution, CPC+Eucaliptol+Juá+Romã+Propolis inhibited the growth of 2 strains at 1/640 dilution and all 28 strains at 1/320 dilution, and Cepacol® showed antimicrobial activity against 3 strains at 1/320 dilution and against all 28 strains at 1/160 dilution. Data were submitted to Kruskal-Wallis test, showing that the MID of Cepacol® was lower than that determined for the other products (p<0.05). In conclusion, CPC-mouthwashes showed antimicrobial activity against S. aureus and the addition of other substances to CPC improved its antimicrobial effect. PMID:19089260

  1. Application of speciated isotope dilution mass spectrometry to evaluate extraction methods for determining mercury speciation in soils and sediments.

    PubMed

    Rahman, G M Mizanur; Kingston, H M Skip

    2004-07-01

    Extraction techniques commonly used to extract methylmercury or mercury species from various matrixes have been evaluated regarding their potential to transform inorganic mercury to methylmercury, or vice versa, during sample preparation steps by applying speciated isotope dilution mass spectrometry. Two of the five tested methods were highly prone to form inorganic mercury from methylmercury. Some published methods converted methylmercury to inorganic mercury approximately 100% (including the spiked CH(3)(201)Hg(+)). In other methods, as much as 45% of methylmercury was converted to inorganic mercury during extraction. The methods evaluated included cold acid extraction and sonication. Other methods, such as the proposed EPA RCRA Draft Method 3200, microwave-assisted extraction, and another sonication-based methods induced very little or no methylmercury transformation to inorganic mercury. Among these three methods, the proposed Draft EPA Method 3200 was found to be the most efficient.

  2. The Soft Agar Colony Formation Assay

    PubMed Central

    Borowicz, Stanley; Van Scoyk, Michelle; Avasarala, Sreedevi; Karuppusamy Rathinam, Manoj Kumar; Tauler, Jordi; Bikkavilli, Rama Kamesh; Winn, Robert A.

    2014-01-01

    Anchorage-independent growth is the ability of transformed cells to grow independently of a solid surface, and is a hallmark of carcinogenesis. The soft agar colony formation assay is a well-established method for characterizing this capability in vitro and is considered to be one of the most stringent tests for malignant transformation in cells. This assay also allows for semi-quantitative evaluation of this capability in response to various treatment conditions. Here, we will demonstrate the soft agar colony formation assay using a murine lung carcinoma cell line, CMT167, to demonstrate the tumor suppressive effects of two members of the Wnt signaling pathway, Wnt7A and Frizzled-9 (Fzd-9). Concurrent overexpression of Wnt7a and Fzd-9 caused an inhibition of colony formation in CMT167 cells. This shows that expression of Wnt7a ligand and its Frizzled-9 receptor is sufficient to suppress tumor growth in a murine lung carcinoma model. PMID:25408172

  3. Dilution space ratio of 2H and 18O of doubly labeled water method in humans.

    PubMed

    Sagayama, Hiroyuki; Yamada, Yosuke; Racine, Natalie M; Shriver, Timothy C; Schoeller, Dale A

    2016-06-01

    Variation of the dilution space ratio (Nd/No) between deuterium ((2)H) and oxygen-18 ((18)O) impacts the calculation of total energy expenditure (TEE) by doubly labeled water (DLW). Our aim was to examine the physiological and methodological sources of variation of Nd/No in humans. We analyzed data from 2,297 humans (0.25-89 yr old). This included the variables Nd/No, total body water, TEE, body mass index (BMI), and percent body fat (%fat). To differentiate between physiologic and methodologic sources of variation, the urine samples from 54 subjects were divided and blinded and analyzed separately, and repeated DLW dosing was performed in an additional 55 participants after 6 mo. Sex, BMI, and %fat did not significantly affect Nd/No, for which the interindividual SD was 0.017. The measurement error from the duplicate urine sample sets was 0.010, and intraindividual SD of Nd/No in repeats experiments was 0.013. An additional SD of 0.008 was contributed by calibration of the DLW dose water. The variation of measured Nd/No in humans was distributed within a small range and measurement error accounted for 68% of this variation. There was no evidence that Nd/No differed with respect to sex, BMI, and age between 1 and 80 yr, and thus use of a constant value is suggested to minimize the effect of stable isotope analysis error on calculation of TEE in the DLW studies in humans. Based on a review of 103 publications, the average dilution space ratio is 1.036 for individuals between 1 and 80 yr of age. PMID:26989221

  4. Comparing the mannitol-egg yolk-polymyxin agar plating method with the three-tube most-probable-number method for enumeration of Bacillus cereus spores in raw and high-temperature, short-time pasteurized milk.

    PubMed

    Harper, Nigel M; Getty, Kelly J K; Schmidt, Karen A; Nutsch, Abbey L; Linton, Richard H

    2011-03-01

    The U.S. Food and Drug Administration's Bacteriological Analytical Manual recommends two enumeration methods for Bacillus cereus: (i) standard plate count method with mannitol-egg yolk-polymyxin (MYP) agar and (ii) a most-probable-number (MPN) method with tryptic soy broth (TSB) supplemented with 0.1% polymyxin sulfate. This study compared the effectiveness of MYP and MPN methods for detecting and enumerating B. cereus in raw and high-temperature, short-time pasteurized skim (0.5%), 2%, and whole (3.5%) bovine milk stored at 4°C for 96 h. Each milk sample was inoculated with B. cereus EZ-Spores and sampled at 0, 48, and 96 h after inoculation. There were no differences (P > 0.05) in B. cereus populations among sampling times for all milk types, so data were pooled to obtain overall mean values for each treatment. The overall B. cereus population mean of pooled sampling times for the MPN method (2.59 log CFU/ml) was greater (P < 0.05) than that for the MYP plate count method (1.89 log CFU/ml). B. cereus populations in the inoculated milk samples ranged from 2.36 to 3.46 and 2.66 to 3.58 log CFU/ml for inoculated milk treatments for the MYP plate count and MPN methods, respectively, which is below the level necessary for toxin production. The MPN method recovered more B. cereus, which makes it useful for validation research. However, the MYP plate count method for enumeration of B. cereus also had advantages, including its ease of use and faster time to results (2 versus 5 days for MPN).

  5. Performance of the EUCAST disk diffusion method, the CLSI agar screen method, and the Vitek 2 automated antimicrobial susceptibility testing system for detection of clinical isolates of Enterococci with low- and medium-level VanB-type vancomycin resistance: a multicenter study.

    PubMed

    Hegstad, Kristin; Giske, Christian G; Haldorsen, Bjørg; Matuschek, Erika; Schønning, Kristian; Leegaard, Truls M; Kahlmeter, Gunnar; Sundsfjord, Arnfinn

    2014-05-01

    Different antimicrobial susceptibility testing methods to detect low-level vancomycin resistance in enterococci were evaluated in a Scandinavian multicenter study (n=28). A phenotypically and genotypically well-characterized diverse collection of Enterococcus faecalis (n=12) and Enterococcus faecium (n=18) strains with and without nonsusceptibility to vancomycin was examined blindly in Danish (n=5), Norwegian (n=13), and Swedish (n=10) laboratories using the EUCAST disk diffusion method (n=28) and the CLSI agar screen (n=18) or the Vitek 2 system (bioMérieux) (n=5). The EUCAST disk diffusion method (very major error [VME] rate, 7.0%; sensitivity, 0.93; major error [ME] rate, 2.4%; specificity, 0.98) and CLSI agar screen (VME rate, 6.6%; sensitivity, 0.93; ME rate, 5.6%; specificity, 0.94) performed significantly better (P=0.02) than the Vitek 2 system (VME rate, 13%; sensitivity, 0.87; ME rate, 0%; specificity, 1). The performance of the EUCAST disk diffusion method was challenged by differences in vancomycin inhibition zone sizes as well as the experience of the personnel in interpreting fuzzy zone edges as an indication of vancomycin resistance. Laboratories using Oxoid agar (P<0.0001) or Merck Mueller-Hinton (MH) agar (P=0.027) for the disk diffusion assay performed significantly better than did laboratories using BBL MH II medium. Laboratories using Difco brain heart infusion (BHI) agar for the CLSI agar screen performed significantly better (P=0.017) than did those using Oxoid BHI agar. In conclusion, both the EUCAST disk diffusion and CLSI agar screening methods performed acceptably (sensitivity, 0.93; specificity, 0.94 to 0.98) in the detection of VanB-type vancomycin-resistant enterococci with low-level resistance. Importantly, use of the CLSI agar screen requires careful monitoring of the vancomycin concentration in the plates. Moreover, disk diffusion methodology requires that personnel be trained in interpreting zone edges.

  6. Low bandgap a-Si:H film with better stability prepared by RF PECVD method using helium dilution

    SciTech Connect

    Ray, S.; Hazra, S.; Middya, A.R.; Barua, A.K.

    1994-12-31

    Low bandgap a-Si:H films have been prepared by RF PECVD method (13.56 MHz) using helium as diluent to silane gas. Helium dilution and chamber pressure play an important role to reduce optical gap at substrate temperature {approximately}210 C. This highly photosensitive low bandgap material showed less light induced degradation compared to that for normal bandgap a-Si:H material. Single junction solar cell having efficiency 7.2% (1 cm{sup 2} area) and spectral response up to 850 nm have been fabricated using 1.6 eV a-Si:H film.

  7. An alternative staining method for counting red-eared slider turtle (Trachemys scripta) blood cells using crystal violet in cells diluted with 0.45% sodium chloride.

    PubMed

    Tsai, Chyong-Ying; Yu, Jane-Fang; Wang, Yu-Wen; Fan, Pei-Chia; Cheng, Ting-Yu; Wang, Lih-Chiann

    2014-09-01

    Various staining methods are available for reptilian species blood cell quantification. However, these methods have shown inaccurate differentiation limitations. The current study evaluates staining effects and blood cell counting results using an alternative method, counting blood cells diluted with 0.45% sodium chloride solution and stained with crystal violet. Blood samples from 8 red-eared slider turtles (Trachemys scripta) were collected. Red and white blood cell counts were performed using different methods: the unstained method, the Unopette method, Liu stain, and crystal violet method using blood cells diluted in various sodium chloride solution osmolarities. The staining properties and blood cell count results were compared. The crystal violet method using blood cells diluted in 0.45% sodium chloride solution delivered the best staining and counting results among all of the tested methods, with the lowest average coefficient of variance. The proposed method can easily be performed, serving as a feasible method for blood cell counting in chelonians.

  8. Development of traceable precision dynamic dilution method to generate dimethyl sulphide gas mixtures at sub-nanomole per mole levels for ambient measurement.

    PubMed

    Kim, Mi Eon; Kim, Yong Doo; Kang, Ji Hwan; Heo, Gwi Suk; Lee, Dong Soo; Lee, Sangil

    2016-04-01

    Dimethyl sulphide (DMS) is an important compound in global atmospheric chemistry and climate change. Traceable international standards are essential for measuring accurately the long-term global trend in ambient DMS. However, developing accurate gas standards for sub-nanomole per mole (nmol/mol) mole fractions of DMS in a cylinder is challenging, because DMS is reactive and unstable. In this study, a dynamic dilution method that is traceable and precise was developed to generate sub-nmol/mol DMS gas mixtures with a dynamic dilution system based on sonic nozzles and a long-term (>5 years) stable 10 μmol/mol parent DMS primary standard gas mixtures (PSMs). The dynamic dilution system was calibrated with traceable methane PSMs, and its estimated dilution factors were used to calculate the mole fractions of the dynamically generated DMS gas mixtures. A dynamically generated DMS gas mixture and a 6 nmol/mol DMS PSM were analysed against each other by gas chromatography with flame-ionisation detection (GC/FID) to evaluate the dilution system. The mole fractions of the dynamically generated DMS gas mixture determined against a DMS PSM and calculated with the dilution factor agreed within 1% at 6 nmol/mol. In addition, the dynamically generated DMS gas mixtures at various mole fractions between 0.4 and 11.7 nmol/mol were analysed by GC/FID and evaluated for their linearity. The analytically determined mole fractions showed good linearity with the mole fractions calculated with the dilution factors. Results showed that the dynamic dilution method generates DMS gas mixtures ranging between 0.4 nmol/mol and 12 nmol/mol with relative expanded uncertainties of less than 2%. Therefore, the newly developed dynamic dilution method is a promising reference method for generating sub-nmol/mol DMS gas standards for accurate ambient measurements.

  9. Fabrication of Flexible Au/ZnO/ITO/PET Memristor Using Dilute Electrodeposition Method

    NASA Astrophysics Data System (ADS)

    Fauzi, F. B.; Ani, M. H.; Othman, R.; Azhar, A. Z. A.; Mohamed, M. A.; Herman, S. H.

    2015-11-01

    DRAM has been approaching its maximum physical limit due to the demand of smaller size and higher capacity memory resistor. The researchers have discovered the abilities of a memristor, a Non Volatile Memory (NVM) that could overcome the size and capacity obstacles. This paper discussed about the deposition of zinc oxide (ZnO) on indium tin oxide (ITO) coated polyethylene terephthalate (PET) substrate by electrodeposition. Metallic Zn film was deposited on substrates with varying deposition time from 15 to 120 seconds in very dilute zinc chloride (ZnCl2) aqueous and subsequently oxidized at 150 °C to form ZnO/ITO coated PET junction. The deposited thin film was characterized via x-ray diffraction (XRD) and field emission scanning electron microscopy (FESEM). The results from I-V measurement show the deposited ZnO exhibits pinched hysteresis loop. The hysteresis loop becomes smaller with increasing deposition time. The 15 seconds electrodeposition gave the largest hysteresis loop and largest value of resistive switching ratio of 1.067. The result of the synthesized ZnO on the flexible substrate can be one of the alternatives to replace the current memory system as the flexible memory system.

  10. Ice-affected streamflow records using tracer-dilution discharge methods

    USGS Publications Warehouse

    Capesius, J.P.; Sullivan, J.R.; Williams, C.A.; O'Neill, G. B.; ,

    2002-01-01

    Accurate ice-affected streamflow records are difficult to obtain for several reasons. Problems measuring stage, variable backwater conditions, access limitations in wintertime, and problems measuring flowing water under ice cover all contribute to make ice-affected streamflow records less accurate than open-channel streamflow records. The inaccuracy of ice-affected streamflow records is particularly troublesome for small streams where Instream-Flow water rights exist. The Colorado Water Conservation Board uses these water rights to protect in-stream aquatic communities. In January and February 2002, the U.S. Geological Survey, in cooperation with the Colorado Water Conservation Board, conducted an experiment using a sodium chloride tracer to determine streamflow under ice cover. The purpose of this study is to determine the usefulness and accuracy of ice-affected streamflow records using a sodium chloride tracer that was automatically injected into the stream. The tracer was injected at two gaging stations once per day for up to 25 days. Multiple-parameter water-quality sensors at the two gaging stations monitored background and peak tracer concentrations and conductance. These data were used to determine discharge at each site. A comparison of current-meter measurements to tracer-dilution discharge measurements shows an underestimation of discharge due to inaccuracy of current-meter measurements with ice cover and inconsistent tracer-pump rates caused by partial freezing of the tracer solution in the injection lines.

  11. Investigation of electrolyte measurement in diluted whole blood using spectroscopic and chemometric methods

    NASA Technical Reports Server (NTRS)

    Soller, Babs R.; Favreau, Janice; Idwasi, Patrick O.

    2003-01-01

    The feasibility of using near-infrared (NIR) spectroscopy in combination with partial least-squares (PLS) regression was explored to measure electrolyte concentration in whole blood samples. Spectra were collected from diluted blood samples containing randomized, clinically relevant concentrations of Na+, K+, and Ca2+. Sodium was also studied in lysed blood. Reference measurements were made from the same samples using a standard clinical chemistry instrument. Partial least squares (PLS) was used to develop calibration models for each ion with acceptable results (Na+, R2 = 0.86, CVSEP = 9.5 mmol/L; K+, R2 = 0.54, CVSEP = 1.4 mmol/L; Ca2+, R2 = 0.56, CVSEP = 0.18 mmol/L). Slightly improved results were obtained using a narrower wavelength region (470-925 nm) where hemoglobin, but not water, absorbed indicating that ionic interaction with hemoglobin is as effective as water in causing measurable spectral variation. Good models were also achieved for sodium in lysed blood, illustrating that cell swelling, which is correlated with sodium concentration, is not required for calibration model development.

  12. Use of a {sup 15}N isotope dilution method to assess contaminant effects on soil nitrification

    SciTech Connect

    Nason, G.E.; Dinwoodie, G.D.

    1995-12-31

    Ecologically relevant bioassays are needed to assess effects of contaminants on soil processes such as decomposition and nutrient cycling. This study was conducted to assess the potential of a soil-based nitrification bioassay. Soil samples adjusted to 0.03 MPa moisture content were amended with 0.1, 1.0, 10 and 100 mg kg{sup {minus}1} PCP or PCB, and 0.05, 0.5, 5 and 50 mg kg{sup {minus}1} Hg and preincubated for 7 days. A 2-d incubation was then started by addition of 10 mg kg{sup {minus}1} {sup 15}NO{sub 3}-N. Diethyl ether used as a carrier for PCP addition had little effect on inorganic nitrogen concentrations during the incubation. Net nitrogen mineralization and nitrification were unaffected by PCB. Higher amendment levels of both PCP and Hg resulted in increases in ammonium concentrations and decreases in net nitrification. {sup 15}N-nitrate pool dilution was sensitive to contamination and showed some gross nitrification was occurring even when net nitrification had ceased. Recoveries of Hg and PCB at the end of the study were greater than 90%. Recovery of PCP was 5%. Incubations carried out under sterile and non-sterile conditions indicated that both sorption and biological degradation were factors in the low PCP recovery.

  13. Evaluation of minimal inhibitory concentration of two new materials using tube dilution method: An in vitro study

    PubMed Central

    Hiremath, Geeta S.; Kulkarni, Raghavendra D.; Naik, Balaram D.

    2015-01-01

    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. PMID:25829698

  14. As a bacterial culture medium, citrated sheep blood agar is a practical alternative to citrated human blood agar in laboratories of developing countries.

    PubMed

    Russell, F M; Biribo, S S N; Selvaraj, G; Oppedisano, F; Warren, S; Seduadua, A; Mulholland, E K; Carapetis, J R

    2006-09-01

    Human blood agar (HuBA) is widely used in developing countries for the isolation of bacteria from clinical specimens. This study compared citrated sheep blood agar (CSBA) and HuBA with defibrinated horse blood agar and defibrinated sheep blood agar (DSBA) for the isolation and antibiotic susceptibility testing of reference and clinical strains of Streptococcus pneumoniae, Streptococcus pyogenes, and Staphylococcus aureus. Reference and clinical strains of all organisms were diluted in brain heart infusion and a clinical specimen of cerebrospinal fluid and cultured on all agars. Viable counts, colony morphology, and colony size were recorded. Susceptibility testing for S. pneumoniae and S. pyogenes was performed on defibrinated sheep blood Mueller-Hinton agar, citrated sheep blood Mueller-Hinton agar (CSB MHA), and human blood Mueller-Hinton agar plates. For all organisms, the colony numbers were similar on all agars. Substantially smaller colony sizes and absent or minimal hemolysis were noted on HuBA for all organisms. Antibiotic susceptibility results for S. pneumoniae were similar for the two sheep blood agars; however, larger zone sizes were displayed on HuBA, and quality control for the reference strain failed on HuBA. For S. pyogenes, larger zone sizes were demonstrated on HuBA and CSBA than on DSBA. Poor hemolysis made interpretation of the zone sizes difficult on HuBA. CSBA is an acceptable alternative for the isolation of these organisms. The characteristic morphology is not evident, and hemolysis is poor on HuBA; and so HuBA is not recommended for use for the isolation or the susceptibility testing of any of these organisms. CSB MHA may be suitable for use for the susceptibility testing of S. pneumoniae.

  15. Quantification of Nitrous Oxide from Fugitive Emissions by Tracer Dilution Method using a Mobile Real-time Nitrous Oxide Analyzer

    NASA Astrophysics Data System (ADS)

    Mønster, J.; Rella, C.; Jacobson, G. A.; He, Y.; Hoffnagle, J.; Scheutz, C.

    2012-12-01

    Nitrous oxide is a powerful greenhouse gas considered 298 times stronger than carbon dioxide on a hundred years term (Solomon et al. 2007). The increasing global concentration is of great concern and is receiving increasing attention in various scientific and industrial fields. Nitrous oxide is emitted from both natural and anthropogenic sources. Inventories of source specific fugitive nitrous oxide emissions are often estimated on the basis of modeling and mass balance. While these methods are well-developed, actual measurements for quantification of the emissions can be a useful tool for verifying the existing estimation methods as well as providing validation for initiatives targeted at lowering unwanted nitrous oxide emissions. One approach to performing such measurements is the tracer dilution method (Galle et al. 2001), in which a tracer gas is released at the source location at a known flow. The ratio of downwind concentrations of both the tracer gas and nitrous oxide gives the ratios of the emissions rates. This tracer dilution method can be done with both stationary and mobile measurements; in either case, real-time measurements of both tracer and analyte gas is required, which places high demands on the analytical detection method. To perform the nitrous oxide measurements, a novel, robust instrument capable of real-time nitrous oxide measurements has been developed, based on cavity ring-down spectroscopy and operating in the near-infrared spectral region. We present the results of the laboratory and field tests of this instrument in both California and Denmark. Furthermore, results are presented from measurements using the mobile plume method with a tracer gas (acetylene) to quantify the nitrous oxide and methane emissions from known sources such as waste water treatment plants and composting facilities. Nitrous oxide (blue) and methane (yellow) plumes downwind from a waste water treatment facility.

  16. Dynamic determination of kinetic parameters for the interaction between polypeptide hormones and cell-surface receptors in the perfused rat liver by the multiple-indicator dilution method

    SciTech Connect

    Sato, H.; Sugiyama, Y.; Sawada, Y.; Iga, T.; Sakamoto, S.; Fuwa, T.; Hanano, M. )

    1988-11-01

    Hepatic elimination of epidermal growth factor (EGF) via receptor-mediated endocytosis was studied by a multiple-indicator dilution method in the isolated perfused rat liver, in which cell polarity and spatial organization are maintained. In this method EGF was given with inulin, an extracellular reference, as a bolus into the portal vein, and dilution curves of both compounds in the hepatic vein effluent were analyzed. Analysis of the dilution curve for EGF, compared with that for somatostatin, which showed no specific binding to isolated liver plasma membranes, resulted as follows: (i) both extraction ratio and distribution volume of {sup 125}I-labeled EGF decreased as the injected amount of unlabeled EGF increased; (ii) the ratio plot of the dilution curve for EGF exhibited an upward straight line initially for a short period of time, whereas the ratio plot of somatostatin gradually decreased. The multiple-indicator dilution method was used for other peptides also. Insulin and glucagon, known to have hepatocyte receptors, behaved similarly to EGF in shape of their ratio plots. The kinetic parameters calculated by this analysis were comparable with reported values obtained by in vitro direct binding measurements at equilibrium using liver homogenates. They conclude that the multiple-indicator dilution method is a good tool for analyzing the dynamics of peptide hormones-cell-surface receptor interaction under a condition in which spatial architecture of the liver is maintained.

  17. "Dilute-and-shoot" triple parallel mass spectrometry method for analysis of vitamin D and triacylglycerols in dietary supplements.

    PubMed

    Byrdwell, William Craig

    2011-12-01

    A method is demonstrated for analysis of vitamin D fortified dietary supplements that eliminates virtually all chemical pretreatment prior to analysis, which is referred to as a "dilute-and-shoot" method. Three mass spectrometers, in parallel, plus a UV detector, an evaporative light-scattering detector (ELSD), and a corona charged aerosol detector (CAD) were used to allow a comparison of six detectors simultaneously. Ultraviolet data were analyzed using internal standard, external standard, and response factor approaches. The contents of gelcaps that contained 2,000 IU (50 μg) vitamin D(3) in rice bran oil, diluted to 100 mL, were analyzed without the need for lengthy saponification and extraction. Vitamin D(3) was analyzed using UV detection, extracted ion chromatograms, selected ion monitoring (SIM) atmospheric pressure chemical ionization mass spectrometry (APCI-MS), and two transitions of multiple reaction monitoring (MRM) APCI-MS. The internal standard, external standard, and response factor methods gave values of 0.5870 ± 0.0045, 0.5893 ± 0.0041, and 0.5889 ± 0.0045 μg/mL, respectively, by UV detection. The values obtained by MS were 0.6117 ± 0.0140, 0.6018 ± 0.0244, and 0.5848 ± 0.0146 μg/mL by SIM and two transitions of MRM, respectively. The triacylglycerols in the oils were analyzed using full-scan APCI-MS, electrospray ionization (ESI) MS, up to MS(4), an ELSD, and a CAD. The method proved to be very sensitive for vitamin D(3), as well as triacylglycerols (TAGs), allowing identification of intact TAGs containing fatty acids up to 28 carbons in length. LC-ESI-MS of glycerin polymers is also demonstrated. PMID:21964772

  18. Contribution of the finite volume point dilution method for measurement of groundwater fluxes in a fractured aquifer.

    PubMed

    Jamin, P; Goderniaux, P; Bour, O; Le Borgne, T; Englert, A; Longuevergne, L; Brouyère, S

    2015-11-01

    Measurement of groundwater fluxes is the basis of all hydrogeological study, from hydraulic characterization to the most advanced reactive transport modeling. Usual groundwater flux estimation with Darcy's law may lead to cumulated errors on spatial variability, especially in fractured aquifers where local direct measurement of groundwater fluxes becomes necessary. In the present study, both classical point dilution method (PDM) and finite volume point dilution method (FVPDM) are compared on the fractured crystalline aquifer of Ploemeur, France. The manipulation includes the first use of the FVPDM in a fractured aquifer using a double packer. This configuration limits the vertical extent of the tested zone to target a precise fracture zone of the aquifer. The result of this experiment is a continuous monitoring of groundwater fluxes that lasted for more than 4 days. Measurements of groundwater flow rate in the fracture (Q(t)) by PDM provide good estimates only if the mixing volume (V(w)) (volume of water in which the tracer is mixed) is precisely known. Conversely, the FVPDM allows for an independent estimation of V(w) and Q(t), leading to better precision in case of complex experimental setup such as the one used. The precision of a PDM does not rely on the duration of the experiment while a FVPDM may require long experimental duration to guarantees a good precision. Classical PDM should then be used for rapid estimation of groundwater flux using simple experimental setup. On the other hand, the FVPDM is a more precise method that has a great potential for development but may require longer duration experiment to achieve a good precision if the groundwater fluxes investigated are low and/or the mixing volume is large. PMID:26519822

  19. Contribution of the finite volume point dilution method for measurement of groundwater fluxes in a fractured aquifer.

    PubMed

    Jamin, P; Goderniaux, P; Bour, O; Le Borgne, T; Englert, A; Longuevergne, L; Brouyère, S

    2015-11-01

    Measurement of groundwater fluxes is the basis of all hydrogeological study, from hydraulic characterization to the most advanced reactive transport modeling. Usual groundwater flux estimation with Darcy's law may lead to cumulated errors on spatial variability, especially in fractured aquifers where local direct measurement of groundwater fluxes becomes necessary. In the present study, both classical point dilution method (PDM) and finite volume point dilution method (FVPDM) are compared on the fractured crystalline aquifer of Ploemeur, France. The manipulation includes the first use of the FVPDM in a fractured aquifer using a double packer. This configuration limits the vertical extent of the tested zone to target a precise fracture zone of the aquifer. The result of this experiment is a continuous monitoring of groundwater fluxes that lasted for more than 4 days. Measurements of groundwater flow rate in the fracture (Q(t)) by PDM provide good estimates only if the mixing volume (V(w)) (volume of water in which the tracer is mixed) is precisely known. Conversely, the FVPDM allows for an independent estimation of V(w) and Q(t), leading to better precision in case of complex experimental setup such as the one used. The precision of a PDM does not rely on the duration of the experiment while a FVPDM may require long experimental duration to guarantees a good precision. Classical PDM should then be used for rapid estimation of groundwater flux using simple experimental setup. On the other hand, the FVPDM is a more precise method that has a great potential for development but may require longer duration experiment to achieve a good precision if the groundwater fluxes investigated are low and/or the mixing volume is large.

  20. Dilution Confusion: Conventions for Defining a Dilution

    ERIC Educational Resources Information Center

    Fishel, Laurence A.

    2010-01-01

    Two conventions for preparing dilutions are used in clinical laboratories. The first convention defines an "a:b" dilution as "a" volumes of solution A plus "b" volumes of solution B. The second convention defines an "a:b" dilution as "a" volumes of solution A diluted into a final volume of "b". Use of the incorrect dilution convention could affect…

  1. [Determining the parameters for receptor-ligand interaction by serial dilution method for the case when the ligand and receptor are in a pre-existing mixture].

    PubMed

    Bobrovnik, S A

    2005-01-01

    New methods of determining the binding parameters for ligand-receptor interaction are considered. The considered approaches are based on the earlier suggested method of serial dilution and application of so-called coordinates of dilution. It was shown that the suggested methods allow to evaluate affinity constant and ligand concentration even for the case, when the receptor and corresponding ligand of unknown concentration are in a mixture and their separation from each other is impossible. In this connection the suggested methods are especially useful for studying the ligand-receptor interaction if the receptor is very liable and its purification from the ligand would cause drastic changes of its binding properties.

  2. A NEW SELECTIVE BLOOD AGAR MEDIUM FOR STREPTOCOCCUS PYOGENES AND OTHER HAEMOLYTIC STREPTOCOCCI.

    PubMed

    LOWBURY, E J; KIDSON, A; LILLY, H A

    1964-05-01

    Horse blood agar containing polymyxin B sulphate, neomycin sulphate, and fusidic acid inhibited the growth of Staph. aureus, Ps. pyocyanea, Proteus mirabilis, E. coli, and Klebsiella pneumoniae but allowed good growth of, and haemolysis by, Str. pyogenes. In a comparison with blood agar, blood 4% agar, and gentian violet blood agar, the selective medium (P.N.F.) yielded a significantly higher proportion of streptococci than the other media, both by aerobic and by anaerobic culture, from burn swab extracts deliberately contaminated with Str. pyogenes; P.N.F. culture was more effective with dilute than with heavy inocula of Str. pyogenes, allowing from three to seven times as many recoveries of Str. pyogenes from swabs contaminated with 10(-3) dilution of streptococcal cultures than cultures of the same material on the other media. Haemolytic streptococci of groups A, C, D, G, and others were isolated by aerobic culture from burns in a consecutive series of 1,277 swabs more often on P.N.F. medium than on blood 4% agar. Viable counts of 12 strains (including 11 different serological types) of Str. pyogenes showed some reduction in the numbers of colonies compared with counts on blood agar, and some strains grew more slowly on P.N.F. medium. These limitations, however, were offset and outweighed by the higher final yield of streptococci on the selective medium.

  3. Determination of niobium in rocks by an isotope dilution spectrophotometric method

    USGS Publications Warehouse

    Greenland, L.P.; Campbell, E.Y.

    1970-01-01

    Rocks and minerals are fused with sodium peroxide in the presence of carrierfree 95Nb. The fusion cake is leached with water and the precipitate dissolved in hydrofluoric-sulfuric acid mixture. Niobium is extracted into methyl isobutyl ketone and further purified by ion exchange. The amount of niobium is determined spectrophotometrically with 4-(2-pyridylazo)-resorcinol, and the chemical yield of the separations determined by counting 95Nb. This procedure is faster and less sensitive to interferences than previously proposed methods for determining niobium in rocks.The high purity of the separated niobium makes the method applicable to nearly all matrices. ?? 1970.

  4. Agar Diffusion Procedures for Susceptibility Testing of Malassezia pachydermatis: Evaluation of Mueller-Hinton Agar Plus 2 % Glucose and 0.5 µg/ml Methylene Blue as the Test Medium.

    PubMed

    Pasquetti, M; Chiavassa, E; Tizzani, P; Danesi, P; Peano, A

    2015-10-01

    Aim of this study was to verify whether Mueller-Hinton agar supplemented with 2 % glucose and methylene blue (MH-GM), which is used for disk diffusion susceptibility testing of Candida species by the Clinical and Laboratory Standards Institute, is suitable for testing Malassezia pachydermatis. A variant of the disk diffusion procedure utilizing a 9-mm tablet was used to test 31 isolates against clotrimazole and miconazole using MH-GM as test medium. The MH-GM agar optimally supported the growth of all M. pachydermatis isolates, provided that the yeast inoculum was prepared with a lipid source (Tween 40 and 80). Zone edges were frequently definite and clear, facilitating the measurement of zone size and minimizing subjectivity. The inhibition zones correlated with MIC values obtained in a broth dilution assay. The agar diffusion method with MH-GM as the test medium appears as a suitable procedure for testing the susceptibility of M. pachydermatis to CTZ and MCZ in clinical laboratories. This test format may allow processing a large number of isolates in epidemiological studies. This may in turn facilitate clarifying to what extent the problem "drug resistance" accounts for cases of treatment failure in dogs with Malassezia otitis and dermatitis. PMID:26138434

  5. Crystal formation in furunculosis agar

    USGS Publications Warehouse

    Bullock, G.L.; Ross, A.J.

    1964-01-01

    SINCE ITS INTRODUCTION SOME MONTHS AGO, FURUNCULOSIS AGAR has been employed in the diagnosis of suspect furunculosis and also as a general purpose medium. During our work with this medium we have noticed discrete "colonies," of crystalline material, which very closely resemble microbial colonies. These crystal colonies are compact and appear on both the surface and subsurface; they occur in inoculated slants and plates incubated for long periods (2 to 3 weeks), as well as in uninoculated stored medium. As the crystal colonies could be confusing to workers using this medium, we decided to attempt to identify them and also to determine whether storage conditions and different lots of medium affect crystal formation.

  6. Replica plating of colonies from Listeria-selective agars to blood agar to improve the isolation of Listeria monocytogenes from foods.

    PubMed

    Cassiday, P K; Graves, L M; Swaminathan, B

    1990-07-01

    Bacterial colonies from Listeria-selective agars were replica plated to sheep blood agar to screen for beta-hemolysis. By using the replica plating method to test for the beta-hemolytic characteristic of all the colonies growing on Listeria-selective agars instead of picking 3 to 10 suspected colonies for further testing, we recovered Listeria monocytogenes from 59 of 142 Listeria-selective agar plates which contained colonies of hemolytic and nonhemolytic Listeria species and were negative when tested by conventional colony picks.

  7. Method for control of NOx emission from combustors using fuel dilution

    SciTech Connect

    Schefer, Robert W.; Keller, Jay O

    2007-01-16

    A method of controlling NOx emission from combustors. The method involves the controlled addition of a diluent such as nitrogen or water vapor, to a base fuel to reduce the flame temperature, thereby reducing NOx production. At the same time, a gas capable of enhancing flame stability and improving low temperature combustion characteristics, such as hydrogen, is added to the fuel mixture. The base fuel can be natural gas for use in industrial and power generation gas turbines and other burners. However, the method described herein is equally applicable to other common fuels such as coal gas, biomass-derived fuels and other common hydrocarbon fuels. The unique combustion characteristics associated with the use of hydrogen, particularly faster flame speed, higher reaction rates, and increased resistance to fluid-mechanical strain, alter the burner combustion characteristics sufficiently to allow operation at the desired lower temperature conditions resulting from diluent addition, without the onset of unstable combustion that can arise at lower combustor operating temperatures.

  8. Estimation of fluid volumes in hemodialysis patients: comparing bioimpedance with isotopic and dilution methods.

    PubMed

    Piccoli, Antonio

    2014-04-01

    Both single-frequency bioimpedance and multiple-frequency spectroscopy are equally accurate in measuring total-body water and intracellular fluid. Estimates are consistent at a population level but not at the individual level, because of wide limits of agreement. There is no real 'gold standard' method providing estimates with absolute accuracy (in liters). Bioelectrical impedance vector analysis allows comparison of the actual body impedance with that of the reference population (in Ω/m). Hemodialysis prescription can be optimized with the use of this feedback.

  9. Variations on the "dilution" method for reconstituting cytochrome c oxidase into membrane vesicles.

    PubMed

    Ramírez, J; Calahorra, M; Peña, A

    1987-05-15

    A method for the rapid incorporation of cytochrome c oxidase into membranes has been developed. This method essentially consists of obtaining a preparation of the enzyme in which it is isolated and then dissolving it in a medium containing 0.5% of the detergent Tween 20, which gives a final concentration of 0.0125% after reconstitution. These studies revealed an optimal ratio of 1 microgram of enzyme to 5 mg of phospholipids. A similar optimal ratio was found when the amount of protein was varied. The optimum temperature was found to be 30 degrees C. Without a peak value being reached, it was found that the best reconstitution was obtained at pH 7.0-8.0. When measurements were performed either with a fluorescent cyanine (DiSC3) or by the uptake of tetraphenylphosphonium, it was found that the enzyme, with cytochrome c added to the outside, was capable of generating a membrane potential that was negative inside. Using the same procedure, the enzyme could also be reconstituted into vesicles of yeast plasma membrane. The procedure, then, seems adequate for incorporating cytochrome c oxidase into different kinds of membrane vesicles.

  10. Optical absorption of dilute nitride alloys using self-consistent Green’s function method

    PubMed Central

    2014-01-01

    We have calculated the optical absorption for InGaNAs and GaNSb using the band anticrossing (BAC) model and a self-consistent Green’s function (SCGF) method. In the BAC model, we include the interaction of isolated and pair N levels with the host matrix conduction and valence bands. In the SCGF approach, we include a full distribution of N states, with non-parabolic conduction and light-hole bands, and parabolic heavy-hole and spin-split-off bands. The comparison with experiments shows that the first model accounts for many features of the absorption spectrum in InGaNAs; including the full distribution of N states improves this agreement. Our calculated absorption spectra for GaNSb alloys predict the band edges correctly but show more features than are seen experimentally. This suggests the presence of more disorder in GaNSb alloys in comparison with InGaNAs. PMID:24475947

  11. A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity.

    PubMed

    Suh, Jung H; Eltanawy, Abeer; Rangan, Apoorva; Saba, Julie D

    2016-01-01

    A new technique for quantifying sphingosine phosphate lyase activity in biological samples is described. In this procedure, 2-hydrazinoquinoline is used to convert (2E)-hexadecenal into the corresponding hydrazone derivative to improve ionization efficiency and selectivity of detection. Combined utilization of liquid chromatographic separation and multiple reaction monitoring-mass spectrometry allows for simultaneous quantification of the substrate S1P and product (2E)-hexadecenal. Incorporation of (2E)- d5-hexadecenal as an internal standard improves detection accuracy and precision. A simple one-step derivatization procedure eliminates the need for further extractions. Limits of quantification for (2E)-hexadecenal and sphingosine-1-phosphate are 100 and 50fmol, respectively. The assay displays a wide dynamic detection range useful for detection of low basal sphingosine phosphate lyase activity in wild type cells, SPL-overexpressing cell lines, and wild type mouse tissues. Compared to current methods, the capacity for simultaneous detection of sphingosine-1-phosphate and (2E)-hexadecenal greatly improves the accuracy of results and shows excellent sensitivity and specificity for sphingosine phosphate lyase activity detection.

  12. Can the single-breath helium dilution method predict lung volumes as measured by whole-body plethysmography?*

    PubMed Central

    Coertjens, Patrícia Chaves; Knorst, Marli Maria; Dumke, Anelise; Pasqualoto, Adriane Schmidt; Riboldi, João; Barreto, Sérgio Saldanha Menna

    2013-01-01

    OBJECTIVE: To compare TLC and RV values obtained by the single-breath helium dilution (SBHD) method with those obtained by whole-body plethysmography (WBP) in patients with normal lung function, patients with obstructive lung disease (OLD), and patients with restrictive lung disease (RLD), varying in severity, and to devise equations to estimate the SBHD results. METHODS: This was a retrospective cross-sectional study involving 169 individuals, of whom 93 and 49 presented with OLD and RLD, respectively, the remaining 27 having normal lung function. All patients underwent spirometry and lung volume measurement by both methods. RESULTS: TLC and RV were higher by WBP than by SBHD. The discrepancy between the methods was more pronounced in the OLD group, correlating with the severity of airflow obstruction. In the OLD group, the correlation coefficient of the comparison between the two methods was 0.57 and 0.56 for TLC and RV, respectively (p < 0.001 for both). We used regression equations, adjusted for the groups studied, in order to predict the WBP values of TLC and RV, using the corresponding SBHD values. It was possible to create regression equations to predict differences in TLC and RV between the two methods only for the OLD group. The TLC and RV equations were, respectively, ∆TLCWBP-SBHD in L = 5.264 − 0.060 × FEV1/FVC (r2 = 0.33; adjusted r2 = 0.32) and ∆RVWBP-SBHD in L = 4.862 − 0.055 × FEV1/FVC (r2 = 0.31; adjusted r2 = 0.30). CONCLUSIONS: The correction of TLC and RV results obtained by SBHD can improve the accuracy of this method for assessing lung volumes in patients with OLD. However, additional studies are needed in order to validate these equations. PMID:24473761

  13. Application of Microbiological Method Direct Epifluorescence Filter Techique/Aerobic Plate Count Agar in the Identification of Irradiated Herbs and Spices

    PubMed Central

    Di Schiavi, Maria Teresa; Foti, Marina; Mosconi, Maria Cristina; Mattiolo, Giuseppina; Cavallina, Roberta

    2014-01-01

    Irradiation is a preservation technology used to improve the safety and hygienic quality of food. Aim of this study was to assess the applicability and validity of the microbiological screening method direct epifluorescence filter technique (DEFT)/aerobic plate count (APC) (EN 13783:2001) for the identification of irradiated herbs and spices. Tests on non-irradiated and irradiated samples of dried herbs and spices were performed. The method was based on the comparison of APC and count obtained using DEFT. In accordance with the standard reference, this method is not applicable to samples with APC<103 colony forming units (CFU)/g and this is its main limit. The results obtained in our laboratories showed that in 50% of cases of non-irradiated samples and in 96% of the samples treated with ionising radiation, the method was not applicable due to a value of CFU/g <103. PMID:27800348

  14. A method for the routine determination of methylmercury in marine tissue by GC isotope dilution-ICP-MS.

    PubMed

    Valdersnes, Stig; Maage, Amund; Fliegel, Daniel; Julshamn, Kåre

    2012-01-01

    Currently, there is no legal limit for methyl mercury (MeHg) in food; thus, no standardized method for the determination of MeHg in seafood exists within the European jurisdiction. In anticipation of a future legislative limit an inductively coupled plasma isotope dilution mass spectrometry (GC-ICP-ID-MS) method was developed in collaboration with the European Standardization Organization (CEN). The method comprises spiking the tissue sample with Me201Hg, followed by decomposition with tetramethylammonium hydroxide, pH adjustment and derivatization with sodium tetraethylborate, and finally organic extraction of the derivatized MeHg in a hexane phase. Subsequently, the sample is analyzed via GC-ICP-MS and the result calculated using the ID equation. The working range of the method was 0.0005-1.321 mg/kg MeHg in marine tissue, with an internal reproducibility (RSD) of 12-1%. The method was validated based on statistical measures, such as the z-scores, using the commercially available reference materials from National Institute of Standards and Technology Standard Reference Material (NIST SRM) 1566b, NIST SRM 2977 and National Research Council of Canada (NRCC) TORT 2, NRCC, DORM 3, NRCC DOLT 4, and European Reference Material (ERM) CE 464. Z-scores for all standard reference materials, except for NIST SRM 1566b, were better than 11.51. The wide range of marine tissues used during the validation ensures that the method will be applicable for measuring of MeHg in seafood matrixes of all kinds.

  15. Quantifying uncertainty of determination by standard additions and serial dilutions methods taking into account standard uncertainties in both axes.

    PubMed

    Hyk, Wojciech; Stojek, Zbigniew

    2013-06-18

    The analytical expressions for the calculation of the standard uncertainty of the predictor variable either extrapolated or interpolated from a calibration line that takes into account uncertainties in both axes have been derived and successfully verified using the Monte Carlo modeling. These expressions are essential additions to the process of the analyte quantification realized with either the method of standard additions (SAM) or the method of serial dilutions (MSD). The latter one has been proposed as an alternative approach to the SAM procedure. In the MSD approach instead of the sequence of standard additions, the sequence of solvent additions to the spiked sample is performed. The comparison of the calculation results based on the expressions derived to their equivalents obtained from the Monte Carlo simulation, applied to real experimental data sets, confirmed that these expressions are valid in real analytical practice. The estimation of the standard uncertainty of the analyte concentration, quantified via either SAM or MSD or simply a calibration curve, is of great importance for the construction of the uncertainty budget of an analytical procedure. The correct estimation of the standard uncertainty of the analyte concentration is a key issue in the quality assurance in the instrumental analysis.

  16. Comparison of the serial dilution indicator and intragastric titration methods for measurement of meal-stimulated gastric acid secretion in man.

    PubMed

    Hogan, D L; Turken, D; Stern, A I; Isenberg, J I

    1983-11-01

    Two in vivo methods that permit quantitation of gastric acid secretion immediately after the meal are currently in use: intragastric titration and the serial dilution indicator method. During intragastric titration, intragastric pH is artificially maintained at 5.5 to 7 by the continuous addition of alkali to the gastric contents, while during serial dilution the intragastric pH is permitted to seek its natural pH. This study compared gastric acid secretion and serum gastrin in response to a liquid protein meal measured by both techniques in 10 subjects. Mean (+/- SE) 3-hr acid outputs were almost identical (53.6 +/- 6.0 mmol/3 hr with intragastric titration and 52.0 +/- 8.5 mmol/3 hr with serial dilution indicator). Furthermore, 30 min secretory responses in individual subjects were highly correlated (r = 0.98 +/- 0.01, P less than 0.001). Also, in spite of intragastric pH being less than 1.5 by 90 min after the meal during the serial dilution method, total integrated serum gastrin concentrations after the meal were similar (intragastric titration = 20.6 +/- 7.3 ng min/ml versus serial dilution indicator = 23.5 +/- 9.8 ng min/ml) and individual 30-min gastrins during the two separate tests were highly correlated (r = 0.80 +/- 0.06, P less than 0.01). It is concluded that both meal-stimulated gastric acid secretion and serum gastrin concentrations as measured by intragastric titration and by the serial dilution indicator method produced similar results.

  17. An Isotope Dilution Method for High-frequency Measurements of Dissolved Inorganic Carbon concentration in the Surface Ocean

    NASA Astrophysics Data System (ADS)

    Huang, K.; Bender, M. L.; Wanninkhof, R. H.; Cassar, N.

    2013-12-01

    Dissolved inorganic carbon (DIC) is one of the most important species in the ocean carbon system. An autonomous system using isotope dilution as its core method has been developed to obtain high-frequency measurements of dissolved inorganic carbon (DIC) concentrations in the surface ocean. This system accurately mixes a seawater sample and a 13C-labeled sodium bicarbonate solution (spike). The mixed solution is then acidified and sent through a gas permeable membrane contactor. CO2 derived from DIC in the mixture is extracted by a CO2-free gas stream, and is sent to a cavity ring-down spectrometer to analyze its 13C/12C ratio. [DIC] of the seawater can then be derived from the measured 13C/12C, the known mixing ratio and the [DI13C] of the spike. The method has been tested under a wide [DIC] range (1800-2800 μmol/kg) in the laboratory. It has also been deployed on a cruise that surveyed ocean waters to the south of Florida. At a sampling resolution of 4 minutes (15 samples per hour), the relative standard deviation of DIC determined from the laboratory tests and the field deployment is ×0.07% and ×0.09%, respectively. The accuracy of the method is better than 0.1% except where [DIC] varies faster than 5 μmol/kg per minute. Based on the laboratory and field evaluations, we conclude that this method can provide accurate underway [DIC] measurements at high resolution in most oceanic regions. Schematic illustration of the work flow.

  18. Validation of a gas chromatography-mass spectrometry isotope dilution method for the determination of 2-butoxyethanol and other common glycol ethers in consumer products.

    PubMed

    Tokarczyk, Ryszard; Jiang, Ying; Poole, Gary; Turle, Richard

    2010-10-29

    A gas chromatography-mass spectrometry isotope dilution (GC-MS ID) method was developed and tested for the determination of 14 common glycol ethers in consumer products. Stable isotope labelled standards, 2-methoxyethanol-D(7) and 2-butoxyethanol-(13)C(2) (CDN isotopes) were employed to enhance the accuracy and precision of the glycol ethers determination. A 1000-fold sample dilution with methanol was applied to avoid column overload and contamination. At this dilution matrix effects were in most cases negligible and did not interfere with the analysis. The instrument detection limit (IDL) for analysed compounds varied from 0.01 to 1 μg/mL; while the estimated limit of quantification (LoQ) varied between different glycol ethers from 0.02 to 3.4 μg/mL. Calibration was tested in the range of 0.1-200 μg/mL and showed that the linear fit is upheld from 0.1 to 10 μg/mL, and extends beyond this range for some of the analytes. Recoveries of glycol ethers from products with different matrices were similar. The recoveries varied from 87% to 116% between the analysed compounds, while measurements precision varied between 2% and 14%. The method is applicable to products with glycol ether concentrations above 0.002-0.2% (w/w). The concentration range can be extended below the specified limits by decreasing the dilution factor; however, with lower dilution the sample matrix effect is expected to be stronger. Products with very high concentrations of glycol ether (>20%) may need to be further diluted prior to injection to avoid column overload. The method can be used for testing liquid and aerosol products designed for household use, such as cleaners, paints, solvents and paint stripers, for compliance and enforcement of regulations which limit glycol ethers content.

  19. Structure and magnetic properties of Co-doped ZnO dilute magnetic semiconductors synthesized via hydrothermal method

    NASA Astrophysics Data System (ADS)

    Ghosh, K.; Kahol, P. K.; Bhamidipati, S.; Das, N.; Khanra, S.; Wanekaya, A.; Delong, R.

    2012-07-01

    Using X-Ray Diffraction, Scanning Electron Microscopy, and Superconducting Quantum Interference Device magnetometer, detailed structural, morphological, and magnetic properties are reported on undoped and cobalt doped ZnO Dilute Magnetic Semiconductors, which were prepared by the hydrothermal method. Synthesis of undoped ZnO and cobalt-doped ZnO nanorods was carried out using aqueous solutions of Zn(NO3)2ṡ6H2O, Co(C2H3OO)2ṡ4H2O, and NH4OH as hydrolytic catalyst. Samples of different sizes and shapes were synthesized by varying process parameters such as solution molarity (0.05M, 0.15M, 0.3M, 0.5M), pH of the precursors in the range 8-11, growth temperature (100°-130°C), growth time (3-6 hrs), and annealing time. Optimum synthesis parameters to grow ZnO and cobalt-doped ZnO nanorods have been obtained. These nanorods show paramagnetic-like behavior. Our results do not indicate ferromagnetism behavior, unlike reported in thin films and nanocrystalline samples. The differences are likely due to the possible role of nanoparticle size, shape, and different oxygen vacancy concentrations.

  20. Comparison of the BBL CHROMagar Staph aureus Agar Medium to Conventional Media for Detection of Staphylococcus aureus in Respiratory Samples

    PubMed Central

    Flayhart, Diane; Lema, Clara; Borek, Anita; Carroll, Karen C.

    2004-01-01

    Screening for Staphylococcus aureus has become routine in certain patient populations. This study is the first clinical evaluation of the BBL CHROMagar Staph aureus agar (CSA) medium (BD Diagnostics, Sparks, Md.) for detection of S. aureus in nasal surveillance cultures and in respiratory samples from cystic fibrosis (CF) patients. S. aureus colonies appear mauve on CSA. Other organisms are inhibited or produce a distinctly different colony color. S. aureus was identified from all media by slide coagulase, exogenous DNase, and mannitol fermentation assays. Susceptibility testing was performed using the agar dilution method. A total of 679 samples were evaluated. All samples were inoculated onto CSA. Nasal surveillance cultures were inoculated onto sheep blood agar (SBA) (BD Diagnostics), and samples from CF patients were inoculated onto mannitol salt agar (MSA) (BD Diagnostics). Of the 679 samples cultured, 200 organisms produced a mauve color on CSA (suspicious for S. aureus) and 180 were positive for S. aureus on SBA or MSA. Of 200 CSA-positive samples 191 were identified as S. aureus. Nine mauve colonies were slide coagulase negative and were subsequently identified as Staphylococcus lugdunensis (one), Staphylococcus epidermidis (three), Staphylococcus haemolyticus (one), and Corynebacterium species (four). CSA improved the ability to detect S. aureus by recovering 12 S. aureus isolates missed by conventional media. Of the 192 S. aureus isolates recovered, 122 were methicillin susceptible and 70 were methicillin resistant. Overall, the sensitivity and specificity of CSA in this study were 99.5 and 98%, respectively. There was no difference in the performance of the slide coagulase test or in susceptibility testing performed on S. aureus recovered from CSA compared to SBA or MSA. Our data support the use of CSA in place of standard culture media for detection of S. aureus in heavily contaminated respiratory samples. PMID:15297498

  1. Comparison of the BBL CHROMagar Staph aureus agar medium to conventional media for detection of Staphylococcus aureus in respiratory samples.

    PubMed

    Flayhart, Diane; Lema, Clara; Borek, Anita; Carroll, Karen C

    2004-08-01

    Screening for Staphylococcus aureus has become routine in certain patient populations. This study is the first clinical evaluation of the BBL CHROMagar Staph aureus agar (CSA) medium (BD Diagnostics, Sparks, Md.) for detection of S. aureus in nasal surveillance cultures and in respiratory samples from cystic fibrosis (CF) patients. S. aureus colonies appear mauve on CSA. Other organisms are inhibited or produce a distinctly different colony color. S. aureus was identified from all media by slide coagulase, exogenous DNase, and mannitol fermentation assays. Susceptibility testing was performed using the agar dilution method. A total of 679 samples were evaluated. All samples were inoculated onto CSA. Nasal surveillance cultures were inoculated onto sheep blood agar (SBA) (BD Diagnostics), and samples from CF patients were inoculated onto mannitol salt agar (MSA) (BD Diagnostics). Of the 679 samples cultured, 200 organisms produced a mauve color on CSA (suspicious for S. aureus) and 180 were positive for S. aureus on SBA or MSA. Of 200 CSA-positive samples 191 were identified as S. aureus. Nine mauve colonies were slide coagulase negative and were subsequently identified as Staphylococcus lugdunensis (one), Staphylococcus epidermidis (three), Staphylococcus haemolyticus (one), and Corynebacterium species (four). CSA improved the ability to detect S. aureus by recovering 12 S. aureus isolates missed by conventional media. Of the 192 S. aureus isolates recovered, 122 were methicillin susceptible and 70 were methicillin resistant. Overall, the sensitivity and specificity of CSA in this study were 99.5 and 98%, respectively. There was no difference in the performance of the slide coagulase test or in susceptibility testing performed on S. aureus recovered from CSA compared to SBA or MSA. Our data support the use of CSA in place of standard culture media for detection of S. aureus in heavily contaminated respiratory samples.

  2. Residual Agar Determination in Bacterial Spores by Electrospray Ionization Mass Spectrometry

    SciTech Connect

    Wahl, Karen L.; Colburn, Heather A.; Wunschel, David S.; Petersen, Catherine E.; Jarman, Kristin H.; Valentine, Nancy B.

    2010-02-15

    Presented here is an analytical method to detect residual agar from a bacterial spore sample as an indication of culturing on an agar plate. This method is based on the resolubilization of agar polysaccharide from a bacterial spore sample, enzymatic digestion, followed by electrospray ionization tandem mass spectrometry (ESI-MSn) analysis for detection of a specific agar fragment ion. A range of Bacillus species and strains were selected to demonstrate the effectiveness of this approach. The characteristic agar fragment ion was detected in the spores grown on agar that were washed from 1 to 5 times, irradiated or non-irradiated and not in the spores grown in broth. A sample containing approximately 108 spores is currently needed for confident detection of residual agar from culture on agar plates in the presence of bacterial spores with a limit of detection of approximately 1 ppm agar spiked into a broth-grown spore sample. The results of a proficiency test with 42 blinded samples are presented demonstrating the utility of this method with no false positives and only 3 false negatives for samples that were below the detection level of the method as documented.

  3. A stable isotope dilution LC-ESI-MS/MS method for the quantification of pyridoxal-5'-phosphate in whole blood.

    PubMed

    van Zelst, Bertrand D; de Jonge, Robert

    2012-08-15

    Vitamin B6 is a cofactor in numerous biologic processes that include gluconeogenesis, neurotransmitter synthesis and amino acid metabolism. The aim of this study was to develop a method to measure the concentration of the biologically active form of vitamin B6 (pyridoxal-5'-phosphate, PLP) in whole blood with stable isotope dilution LC-ESI-MS/MS and compare this new procedure with an established HPLC method based on derivatization of pyridoxal-5'-phosphate. 50 μl of stable isotope (PLP-d3) was added to 250 μl of sample, followed by deproteinization with 10% trichloroacetic acid. After centrifugation, 20 μl of the supernatant was injected into the LC-ESI-MS/MS. Reversed phase chromatography was performed on a UPLC system, using a Waters™ Symmetry C18 column, with a gradient of 0.1% formic acid in methanol. PLP was measured on a tandem MS with a mass transition of 247.8>149.8 in the positive ion mode with a collision energy of 14 eV. The chromatographic run lasted 4 min. The method was linear from 4 to 8000 nmol/l. The intra-day and inter-day precision ranged between 1.7-2.8% and 3.0-4.1%, respectively. The mean absolute matrix-effect was 99.3% [97-102%]. The relative matrix-effect was 98.8%. The mean recovery was 98% [89-103%]. The lower limit of quantification was 4 nmol/l. The comparison of the LC-ESI-MS/MS method with our current HPLC method yielded the following equation: LC-ESI-MS/MS=1.11 [confidence interval, CI: 1.03-1.20] × HPLC+4.6 [CI: -1.3 to 11.0] (r²=0.94). This LC-ESI-MS/MS based method is characterized by simple sample processing and a short run time. The comparison with the current HPLC method is excellent although a significant proportional bias was detected. To conclude, the LC-ESI-MS/MS method is an appropriate method to determine PLP in whole blood.

  4. Agar polysaccharides from Gracilaria species (Rhodophyta, Gracilariaceae).

    PubMed

    Marinho-Soriano, E

    2001-07-26

    Yield, physical and chemical properties of agar from three agarophytes species (Gracilaria gracilis, G. dura and G. bursa-pastoris) were determined. The agar yield from the three species varied significantly (P<0.01). The highest yields of agar (34.8%) and the lowest (30%) were obtained from G. bursa-pastoris and G. gracilis, respectively. Highest gel strength (630+/-15 g cm(-2)) was obtained from agar extracted from G. gracilis and lowest from G. bursa-pastoris (26+/-3.6 g cm(-2)). The values of 3,6-anhydrogalactose were similar for G. gracilis and G. dura and there were no significant differences among the species. The sulfate contents varied significantly (P<0.01) and the higher value was obtained from G. bursa-pastoris. Among the three species, G. gracilis showed superior agar quality than the other two species, hence it can be considered a good potential source for industrial use.

  5. International experiences in assessing vitamin A status and applying the vitamin A-labeled isotope dilution method.

    PubMed

    Lopez-Teros, Veronica; Chileshe, Justin; Idohou-Dossou, Nicole; Fajarwati, Tetra; Medoua Nama, Gabriel; Newton, Sam; Vinod Kumar, Malavika; Wang, Zhixu; Wasantwisut, Emorn; Hunt, Janet R

    2014-01-01

    Inadequate vitamin A (VA) nutrition continues to be a major problem worldwide, and many interventions being implemented to improve VA status in various populations need to be evaluated. The interpretation of results after an intervention depends greatly on the method selected to assess VA status. To evaluate the effect of an intervention on VA status, researchers in Cameroon, India, Indonesia, Mexico, Senegal and Zambia have used serum retinol as an indicator, and have not always found improvement in response to supplementation. One problem is that homeostatic control of serum retinol may mask positive effects of treatment in that changes in concentration are observed only when status is either moderately to severely depleted or excessive. Because VA is stored mainly in the liver, measurements of hepatic VA stores are the “gold standard” for assessing VA status. Dose response tests such as the relative dose response (RDR) and the modified relative dose response (MRDR), allow a qualitative assessment of VA liver stores. On the other hand, the use of the vitamin A-labeled isotope dilution (VALID) technique, (using 13C or 2H-labeled retinyl acetate) serves as an indirect method to quantitatively estimate total body and liver VA stores. Countries including Cameroon, China, Ghana, Mexico, Thailand and Zambia are now applying the VALID method to sensitively assess changes in VA status during interventions, or to estimate a population’s dietary requirement for VA. Transition to the use of more sensitive biochemical indicators of VA status such as the VALID technique is needed to effectively assess interventions in populations where mild to moderate VA deficiency is more prevalent than severe deficiency. PMID:25537105

  6. Recovery of Sublethally Injured Bacteria Using Selective Agar Overlays.

    ERIC Educational Resources Information Center

    McKillip, John L.

    2001-01-01

    This experiment subjects bacteria in a food sample and an environmental sample to conditions of sublethal stress in order to assess the effectiveness of the agar overlay method to recover sublethally injured cells compared to direct plating onto the appropriate selective medium. (SAH)

  7. Growth kinetics of three species of Tetrahymena on solid agar

    SciTech Connect

    Dobra, K.W.; McArdle, E.W.; Ehret, C.F.

    1980-01-01

    A nutrient-agar method without liquid overlay has been developed for cultivation of ciliates. Three species of Tetrahymena-T. pyriformis strain W, T. rostrata strain UNI, and T. vorax strain V/sub 2/S, representing the 3 main groups of Tetrahymena species, were used; however the method should apply to other ciliates. Growth on the surface of the agar was facilitated by an optimal surface-to-volume ratio yielding a high density of ciliates and short generation times. At the highest density achieved, the cells became irregularly hexagonal and formed a monolayer tissue on the agar. Ciliates grown on agar were like those in liquid culture, typical oral ciliature, food-vacuole formation, and typical cortical patterns being retained. Advantages of this method include high cell density, easy recovery, and optimal O/sub 2/ supply. The organisms can also be cultivated on the surface of sterile cellulose-nitrate filters, facilitating in situ fixation and staining as well as transfer into different media by transfer of filters with cells, without prior centrifugation and resuspension.

  8. Single Strain Isolation Method for Cell Culture-Adapted Hepatitis C Virus by End-Point Dilution and Infection

    PubMed Central

    Sugiyama, Nao; Murayama, Asako; Suzuki, Ryosuke; Watanabe, Noriyuki; Shiina, Masaaki; Liang, T. Jake; Wakita, Takaji; Kato, Takanobu

    2014-01-01

    The hepatitis C virus (HCV) culture system has enabled us to clarify the HCV life cycle and essential host factors for propagation. However, the virus production level of wild-type JFH-1 (JFH-1/wt) is limited, and this leads to difficulties in performing experiments that require higher viral concentrations. As the cell culture-adapted JFH-1 has been reported to have robust virus production, some mutations in the viral genome may play a role in the efficiency of virus production. In this study, we obtained cell culture-adapted virus by passage of full-length JFH-1 RNA-transfected Huh-7.5.1 cells. The obtained virus produced 3 log-fold more progeny viruses as compared with JFH-1/wt. Several mutations were identified as being responsible for robust virus production, but, on reverse-genetics analysis, the production levels of JFH-1 with these mutations did not reach the level of cell culture-adapted virus. By using the single strain isolation method by end-point dilution and infection, we isolated two strains with additional mutations, and found that these strains have the ability to produce more progeny viruses. On reverse-genetics analysis, the strains with these additional mutations were able to produce robust progeny viruses at comparable levels as cell culture-adapted JFH-1 virus. The strategy used in this study will be useful for identifying strains with unique characteristics, such as robust virus production, from a diverse population, and for determining the responsible mutations for these characteristics. PMID:24848954

  9. Achieving comparability with IFCC reference method for the measurement of hemoglobin A1c by use of an improved isotope-dilution mass spectrometry method.

    PubMed

    Liu, Hong; Wong, Lingkai; Yong, Sharon; Liu, Qinde; Lee, Tong Kooi

    2015-10-01

    The development of reference measurement methods for hemoglobin A1c (HbA1c) is important for quality assurance in diabetes management. The IFCC reference method using purified proteins as calibration standards is the recommended accuracy-based reference method for the standardization of HbA1c measurement. We developed a highly precise and accurate liquid chromatography-isotope-dilution tandem mass spectrometry (LC-IDMS/MS) procedure, which can serve as an alternative accuracy-based method for HbA1c measurement. In this method, enzymatic proteolysis was applied to sample preparation, followed by LC-IDMS/MS measurement of hemoglobin A0 (HbA0) and HbA1c, using two "signature" hexapeptides for calibration. The concentrations of the signature hexapeptide calibration solutions were, in turn, determined using a hydrolysis method with HCl, followed by LC-IDMS/MS measurement using amino acid solutions as calibration standards. These solutions were gravimetrically prepared from pure amino acid certified reference materials (CRMs). The developed LC-IDMS/MS method was used in participation in an IFCC ring trial for reference laboratories (RELA 2013 and 2014) for HbA1c, where our results were compared with those using the IFCC reference method. The deviations were found to be 0.4-1.7 mmol mol(-1) [or 0.04-0.16% in National Glygohemoglobin Standardization Program (NGSP) units], revealing good comparability with the IFCC reference method. The relative expanded uncertainty of the LC-IDMS/MS was in the range of 2.6% to 2.8% (1.6% to 2.2% after converting to NGSP units). With excellent method precision, good comparability with the IFCC reference method, and a small measurement uncertainty, the developed LC-IDMS/MS method may be used as an alternative accuracy-based reference method for HbA1c measurement.

  10. Use of a novel radiometric method to assess the inhibitory effect of donepezil on acetylcholinesterase activity in minimally diluted tissue samples

    PubMed Central

    Kikuchi, Tatsuya; Okamura, Toshimitsu; Arai, Takuya; Obata, Takayuki; Fukushi, Kiyoshi; Irie, Toshiaki; Shiraishi, Tetsuya

    2010-01-01

    Background and purpose: Cholinesterase inhibitors have been widely used for the treatment of patients with dementia. Monitoring of the cholinesterase activity in the blood is used as an indicator of the effect of the cholinesterase inhibitors in the brain. The selective measurement of cholinesterase with low tissue dilution is preferred for accurate monitoring; however, the methods have not been established. Here, we investigated the effect of tissue dilution on the action of cholinesterase inhibitors using a novel radiometric method with selective substrates, N-[14C]methylpiperidin-4-yl acetate ([14C]MP4A) and (R)-N-[14C]methylpiperidin-3-yl butyrate ([14C]MP3B_R), for AChE and butyrylcholinesterase (BChE) respectively. Experimental approach: We investigated the kinetics of hydrolysis of [14C]-MP4A and [14C]-MP3B_R by cholinesterases, and evaluated the selectivity of [14C]MP4A and [14C]MP3B_R for human AChE and BChE, respectively, compared with traditional substrates. Then, IC50 values of cholinesterase inhibitors in minimally diluted and highly diluted tissues were measured with [14C]MP4A and [14C]MP3B_R. Key results: AChE and BChE activities were selectively measured as the first-order hydrolysis rates of [14C]-MP4A and [14C]MP3B_R respectively. The AChE selectivity of [14C]MP4A was an order of magnitude higher than traditional substrates used for the AChE assay. The IC50 values of specific AChE and BChE inhibitors, donepezil and ethopropazine, in 1.2-fold diluted human whole blood were much higher than those in 120-fold diluted blood. In addition, the IC50 values of donepezil in monkey brain were dramatically decreased as the tissue was diluted. Conclusions and implications: This method would effectively monitor the activity of cholinesterase inhibitors used for therapeutics, pesticides and chemical warfare agents. PMID:20401964

  11. Bacterial pathogens of otitis media and sinusitis: detection in the nasopharynx with selective agar media.

    PubMed

    Dudley, S; Ashe, K; Winther, B; Hendley, J O

    2001-11-01

    Carriage rates for the bacterial pathogens associated with otitis media (Streptococcus pneumoniae [SP], Hemophilus influenzae [HI], and Moraxella catarrhalis [MC]) are of interest. Culture on three selective agars was compared with culture on two standard agars to determine the more accurate method for detection of these species in the nasopharynx of healthy children. Weekly samples were obtained in winter from 18 healthy children (ages 1 through 9 years) as part of a longitudinal study. A 0.1-mL sample of 116 nasopharyngeal aspirate/washes was inoculated onto each of five agars. Two were standard (sheep blood and chocolate), and three were selective (blood with gentamicin for SP; chocolate with vancomycin, bacitracin, and clindamycin for HI; blood with amphotericin B, vancomycin, trimethoprim, and acetazolamide for MC). One technician read the standard plates and another the selective; both were blinded to the results of the other. SP was found in 44% of samples with selective agar versus 25% with standard agar; HI was found in 31% with selective versus 9% with standard; MC was found in 56% with selective versus 37% with standard. Overall, 80% of samples had one or more pathogens detected with selective agars as compared with 58% with standard agars (P =.0004). Selective agars were more accurate than standard agars for detecting otitis pathogens in the nasopharynx, where they are a common part of normal flora in healthy children.

  12. Determination of thyroid hormones in mouse tissues by isotope-dilution microflow liquid chromatography-mass spectrometry method.

    PubMed

    De Angelis, Meri; Giesert, Florian; Finan, Brian; Clemmensen, Christoffer; Müller, Timo D; Vogt-Weisenhorn, Daniela; Tschöp, Matthias H; Schramm, Karl-Werner

    2016-10-15

    Thyroid hormones (THs) play a critical role in the regulation of many biological processes such as growth, metabolism and development both in humans and wildlife. In general, TH levels are measured by immunoassay (IA) methods but the specificity of the antibodies used in these assays limits selectivity. In the last decade, several analytical methods using liquid chromatography-mass spectrometry (LC-MS) and tandem mass spectrometry (LC-MS/MS) have been developed to measure THs. These new techniques proved to be more accurate than the IA analysis and they were widely used for the determination of TH level in different human and animal tissues. A large part of LC-MS/MS methods described in literature employed between 200 and 500mg of sample, however this quantity can be considered too high especially when preclinical studies are conducted using mice as test subjects. Thus an analytical method that reduces the amount of tissue is essential. In this study, we developed a procedure for the analysis of six THs; L-thyroxine (T4), 3,3',5-triiodo-l-thyronine (T3), 3,3',5'-triiodo-l-thyronine (rT3), 3,5-diiodo-l-thyronine (rT2), 3,3'-diiodo-l-thyronine (T2), 3-iodo-l-thyronine (T1) using isotope ((13)C6-T4, (13)C6-T3, (13)C6-rT3, (13)C6-T2) dilution liquid chromatography-mass spectrometry. The major difference with previously described methods lies in the utilization of a nano-UPLC (Ultra Performance Liquid Chromatography) system in micro configuration. This approach leads to a reduction compared to the published methods, of column internal diameter, flow rate, and injected volume. The result of all these improvements is a decrease in the amount of sample necessary for the analysis. The method was tested on six different mouse tissues: liver, heart, kidney, muscle, lung and brown adipose tissue (BAT). The nano-UPLC system was interfaced with a quadrupole time-of-flight mass spectrometer (Q-TOF2-MS) using the positive ion mode electrospray ionization. In our analytical method

  13. Some Experiments With Agar-Grown Seedlings

    ERIC Educational Resources Information Center

    Freeland, P. W.

    1973-01-01

    Two percent agar gel is reported as a better medium for germination and growth studies. Students can be encouraged to undertake many simple experiments and make precise observations by using this medium. (PS)

  14. Characterization of physicochemical properties of carboxymethyl agar.

    PubMed

    Cao, Mingzhao; Liu, Xin; Luan, Jimei; Zhang, Xiaodong

    2014-10-13

    A series of carboxymethyl agars (CMAs) with different degree of substitution (DS) were prepared, and their properties were determined and analyzed. The results showed that with the increase of DS, the dissolving temperature, the gelling temperature, the gel melting temperature, the gel strength, the gel hardness, the gel fracturability, and the solution apparent viscosity of CMA all decreased, except that its gel cohesiveness and gel springiness increased. The variation process of agar molecules in solution from coil to helix could be observed by measuring the optical rotation of the solution at such a low concentration, at which even the solution could not form a gel. The gel skeleton microstructures of both agar and CMA were of porous network structure, and the pore size of CMA became smaller and denser with the increase of its DS. After carboxymethylation, the agar hygroscopicity was improved, but its thermal stability was lowered.

  15. Procedural revision to the use-dilution methods: establishment of maximum log density value for test microbes on inoculated carriers.

    PubMed

    Tomasino, Stephen F; Pines, Rebecca M; Hamilton, Gordon C

    2012-01-01

    (Staphylococcus aureus) and 964.02 (Pseudomonas aeruginosa), were revised in 2009 to include a standardized procedure to measure the log density of the test microbe and to establish a minimum mean log density value of 6.0 (geometric mean of 1.0 x 10(6) CFU/carrier) to qualify the test results. This report proposes setting a maximum mean log density value of 7.0 (geometric mean of 1.0 x 10(7) CFU/carrier) to further standardize the procedure. The minimum value was based on carrier count data collected by four laboratories over an 8-year period (1999-2006). The data have been updated to include an additional 4 years' worth of data (2006-2010) collected by the same laboratories. A total of 512 tests were conducted on products bearing claims against P. aeruginosa and S. aureus with and without an organic soil load (OSL) added to the inoculum (as specified on the product label claim). Six carriers were assayed in each test, for a total of 3072 carriers. Mean log densities for each of the 512 tests were at least 6.0. With the exception of two tests, one for P. aeruginosa without OSL and one for S. aureus with OSL, the mean log densities did not exceed 7.5 (geometric mean of 3.2 x 10(7) CFU/carrier). Across microbes and OSL treatments, the mean log density (+/- SEM) was 6.80 (+/- 0.07) per carrier (a geometric mean of 6.32 x 10(6) CFUlcarrier) and acceptable repeatability (0.28) and reproducibility (0.31) SDs were exhibited. A maximum mean log density per carrier of 7.0 is being proposed here as a validity requirement for S. aureus and P. aeruginosa. A modification to the method to allow for dilution of the final test cultures to achieve carrier counts within 6.0-7.0 logs is also being proposed. Establishing a range of 6.0-7.0 logs will help improve the reliability of the method and should allow for more consistent results within and among laboratories.

  16. Differentiation of Candida dubliniensis from Candida albicans on rosemary extract agar and oregano extract agar.

    PubMed

    de Loreto, Erico Silva; Pozzatti, Patrícia; Alves Scheid, Liliane; Santurio, Deise; Morais Santurio, Janio; Alves, Sydney Hartz

    2008-01-01

    Candida dubliniensis is a recently described pathogenic species which shares many phenotypic features with Candida albicans and therefore, may be misidentified in microbiological laboratories. Because molecular methods can be onerous and unfeasible in routine mycological laboratories with restricted budgets such as those in developing countries, phenotypic techniques have been encouraged in the development of differential media for the presumptive identification of these species. We examined the colony morphology and chlamydospore production of 30 C. dubliniensis isolates and 100 C. albicans isolates on two new proposed media: rosemary (Rosmarinus officinalis) extract agar (REA) and oregano (Origanum vulgare) extract agar (OEA). These substrates are traditionally used as spices and medicinal herbs. In both of these media, all C. dubliniensis isolates (100%) showed rough colonies with peripheral hyphal fringes and abundant chlamydospores after 24 to 48 hr of incubation at 25 degrees C. In contrast, under the same conditions, all isolates of C. albicans (100%) showed smooth colonies without hyphal fringes or chlamydospores. In conclusion, REA and OEA offer a simple, rapid, and inexpensive screening media for the differentiation of C. albicans and C. dubliniensis.

  17. Agar composition affects in vitro screening of biocontrol activity of antagonistic microorganisms.

    PubMed

    Bosmans, L; De Bruijn, I; De Mot, R; Rediers, H; Lievens, B

    2016-08-01

    Agar-based screening assays are the method of choice when evaluating antagonistic potential of bacterial biocontrol-candidates against pathogens. We showed that when using the same medium, but different agar compositions, the activity of a bacterial antagonist against Agrobacterium was strongly affected. Consequently, results from in vitro screenings should be interpreted cautiously. PMID:27166668

  18. Fusobacterium necrophorum- detection and identification on a selective agar.

    PubMed

    Bank, Steffen; Nielsen, Hanne Merete; Mathiasen, Boris Hoyer; Leth, Dorte Christiansen; Kristensen, Lena Hagelskjaer; Prag, Jørgen

    2010-12-01

    Within the last decade, Fusobacterium necrophorum subsp. funduliforme has been considered a clinically important pathogen causing pharyngitis especially in adolescents and young adults. F. necrophorum pharyngitis can progress into Lemierre's syndrome, which is a severe and life-threatening infection. However, throat swabs are not cultured anaerobically in the routine and even if cultured anaerobically, it can be difficult to identify F. necrophorum from the normal flora of the throat. F. necrophorum is therefore often overlooked as the cause of pharyngitis. In our laboratory, a F. necrophorum selective agar has been developed containing vancomycin and nalidixin, which inhibit the growth of most Gram-positive and many Gram-negative bacteria, respectively. β-haemolysis of horse blood can be detected, which further facilitates the detection and identification of F. necrophorum. The F. necrophorum selective agar was evaluated against a quantitative real-time polymerase chain reaction assay and shown to have a significantly higher sensitivity for detecting F. necrophorum than the anaerobic agar commonly used in Denmark. Furthermore, the F. necrophorum selective agar does not require experienced laboratory technicians, require fewer subcultures, is probably less expensive and is faster to perform than other culture methods.

  19. Methicillin-Resistant Staphylococcus aureus (MRSA) Detection: Comparison of Two Molecular Methods (IDI-MRSA PCR Assay and GenoType MRSA Direct PCR Assay) with Three Selective MRSA Agars (MRSA ID, MRSASelect, and CHROMagar MRSA) for Use with Infection-Control Swabs▿

    PubMed Central

    van Hal, S. J.; Stark, D.; Lockwood, B.; Marriott, D.; Harkness, J.

    2007-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is an increasing problem. Rapid detection of MRSA-colonized patients has the potential to limit spread of the organism. We evaluated the sensitivities and specificities of MRSA detection by two molecular methods (IDI-MRSA PCR assay and GenoType MRSA Direct PCR assay) and three selective MRSA agars (MRSA ID, MRSASelect, and CHROMagar MRSA), using 205 (101 nasal, 52 groin, and 52 axillary samples) samples from consecutive known MRSA-infected and/or -colonized patients. All detection methods had higher MRSA detection rates for nasal swabs than for axillary and groin swabs. Detection of MRSA by IDI-MRSA was the most sensitive method, independent of the site (94% for nasal samples, 80% for nonnasal samples, and 90% overall). The sensitivities of the GenoType MRSA Direct assay and the MRSA ID, MRSASelect, and CHROMagar MRSA agars with nasal swabs were 70%, 72%, 68%, and 75%, respectively. All detection methods had high specificities (95 to 99%), independent of the swab site. Extended incubation for a further 24 h with selective MRSA agars increased the detection of MRSA, with a corresponding decline in specificity secondary to a significant increase in false-positive results. There was a noticeable difference in test performance of the GenoType MRSA Direct assay in detection of MRSA (28/38 samples [74%]) compared with detection of nonmultiresistant MRSA (17/31 samples [55%]) (susceptible to two or more non-β-lactam antibiotics). This was not observed with selective MRSA agar plates or IDI-MRSA. Although it is more expensive, in addition to rapid turnaround times of 2 to 4 h, IDI-MRSA offers greater detection of MRSA colonization, independent of the swab site, than do conventional selective agars and GenoType MRSA Direct. PMID:17537949

  20. Bioscreening and expression of a camel anti-CTGF VHH nanobody and its renaturation by a novel dialysis-dilution method.

    PubMed

    Xue, Xiulei; Fan, Xiaobo; Qu, Qingrong; Wu, Guoqiu

    2016-12-01

    The variable regions of the camel heavy chain antibody, also known as nanobody is the smallest antibody with antigen-binding efficiency. CTGF is considered important during extracellular matrix deposition which was involved in the pathogenesis of fibrosis related diseases. There are several anti-CTGF-C nanobody drugs under developing in pharmacy. In this study, we described the screening of a novel anti-CTGF-C nanobody from the peripheral blood of immunized camel by phage display. The screened nanobody was further expressed and purified from E. coli cells. A sophisticated dialysis-dilution method was designed for the in vitro refolding of the nanobody. The results showed that the expressed nanobody was consisted of 135 amino acid and mainly expressed as inclusion body in E. coli cells. The dialysis-dilution method was very effective and the recovery rate of the renaturation was more than 80 %. The ELISA result suggested the nanobody had been well refolded showing a superior CTGF binding activity to the commercial mouse anti-CTGF-C mAb. In conclusion, the anti-CTGF-C nonobody had been successfully screened by phage display. The dialysis-dilution refolding method was very effective and the recovery rate reached over 80 %. PMID:27620736

  1. Thermodynamics of Dilute Solutions.

    ERIC Educational Resources Information Center

    Jancso, Gabor; Fenby, David V.

    1983-01-01

    Discusses principles and definitions related to the thermodynamics of dilute solutions. Topics considered include dilute solution, Gibbs-Duhem equation, reference systems (pure gases and gaseous mixtures, liquid mixtures, dilute solutions), real dilute solutions (focusing on solute and solvent), terminology, standard states, and reference systems.…

  2. Analysis of pressure drop characteristics and methods for calculating gas and gas-solid flow in horizontal pipes for dilute coal conveying system

    SciTech Connect

    Weiguo Pan; Zuohe Chi; Yongjing Liao

    1997-07-01

    This article reported pressure drop characteristics and methods for calculating friction factors {lambda} 0 and {lambda}{sub {mu}} for gas and gas-solids flows, respectively, in straight horizontal pipes are summarized advantages seed. The and disadvantages of calculating friction factor {lambda}{sub {mu}} through dimensional analysis in comparison with model simulation are analyzed. It is pointed out that model simulation is more suitable to engineering use than dimensional analysis. According to experimental results of dilute gas-coal powder flow in straight horizontal pipes of the coal pulverization system in a power plant; an empirical formula and a theoretical formula for calculating friction factor {lambda}{sub {mu}} in straight horizontal pipes transporting dilute coal powder are obtained.

  3. Agar agar-stabilized milled zerovalent iron particles for in situ groundwater remediation.

    PubMed

    Velimirovic, Milica; Schmid, Doris; Wagner, Stephan; Micić, Vesna; von der Kammer, Frank; Hofmann, Thilo

    2016-09-01

    Submicron-scale milled zerovalent iron (milled ZVI) particles produced by grinding macroscopic raw materials could provide a cost-effective alternative to nanoscale zerovalent iron (nZVI) particles for in situ degradation of chlorinated aliphatic hydrocarbons in groundwater. However, the aggregation and settling of bare milled ZVI particles from suspension presents a significant obstacle to their in situ application for groundwater remediation. In our investigations we reduced the rapid aggregation and settling rate of bare milled ZVI particles from suspension by stabilization with a "green" agar agar polymer. The transport potential of stabilized milled ZVI particle suspensions in a diverse array of natural heterogeneous porous media was evaluated in a series of well-controlled laboratory column experiments. The impact of agar agar on trichloroethene (TCE) removal by milled ZVI particles was assessed in laboratory-scale batch reactors. The use of agar agar significantly enhanced the transport of milled ZVI particles in all of the investigated porous media. Reactivity tests showed that the agar agar-stabilized milled ZVI particles were reactive towards TCE, but that their reactivity was an order of magnitude less than that of bare, non-stabilized milled ZVI particles. Our results suggest that milled ZVI particles could be used as an alternative to nZVI particles as their potential for emplacement into contaminated zone, their reactivity, and expected longevity are beneficial for in situ groundwater remediation.

  4. Standard operating procedure to prepare agar phantoms

    NASA Astrophysics Data System (ADS)

    Souza, R. M.; Santos, T. Q.; Oliveira, D. P.; Souza, R. M.; Alvarenga, A. V.; Costa-Felix, R. P. B.

    2016-07-01

    Agar phantoms are widely used as soft tissue mimics and some preparation techniques are described in the literature. There are also standards that describe the recipe of a soft tissue mimicking material (TMM). However some details of manufacture process are not clearly defined. The standardization of the phantom's preparation can produce a metrological impact on the results of the acoustic properties measured. In this direction, this paper presents a standard operating procedure (SOP) to prepare the agar TMM described on the IEC 60601-237.

  5. [Optimization of HIV diagnosis using the RT-PCR (gag) method with various dilutions of cDNA and MgCl2 molarity].

    PubMed

    Subasić, D; Konjhodzić, R; Serić, K

    1999-01-01

    Lack of optimum conditions for PCR can lead to absence of desired PCR products, undefined multiplication and appearance of unwanted products. So, the use of PCR aiming to generate large amounts of target nucleic acid sequences, may be so called "double-edged sword". The important parameters in optimisation of PCR methodology are annealing temperatures, Mg++ concentration and different dilutions of target sequences. In our optimization experiments of HIV-RT-PCR (GAG) method we used HIV positive plasma specimens for extraction of RNA and production of cDNA by reverse transcriptase. Different cDNA dilution (10(-1)-10(-10)) and MgCl2 molarity (1.25 mM; 1.5 mM; 5.0 mM) we used for first round (GAG1 and GAG4 outer primers) and second round PCR (GAG2 and GAG3 inner primers). Optimal results after 3% NuSieve agarose gel electrophoresis and detection of 413 pb PCR products were obtained with 1.25 mM MgCl2 and cDNA dilution 10(-1) and 10(-2). So the main aim of PCR optimisation is the achievement of optimal primer template binding and primer extension.

  6. Use of agar agar stabilized milled zero-valent iron particles for in situ groundwater remediation

    NASA Astrophysics Data System (ADS)

    Schmid, Doris; Velimirović, Milica; Wagner, Stephan; Micić Batka, Vesna; von der Kammer, Frank; Hofmann, Thilo

    2015-04-01

    A major obstacle for use of nanoscale zero-valent iron (nZVI) particles as a nontoxic material for effective in situ degradation of chlorinated aliphatic hydrocarbons (CAHs) is the high production cost. For that reason, submicro-scale milled zero-valent iron particles were recently developed (milled ZVI, UVR-FIA, Germany) by grinding macroscopic raw materials of elementary iron as a cheaper alternative to products produced by solid-state reduction. However, milled ZVI particles tend to aggregate and due to the rather large particle size (d50= 11.9 µm) also rapidly sediment. To prevent aggregation and consequently sedimentation of milled ZVI particles and therefore improve the mobility after in situ application, the use of a stabilizer is considered in literature as a most promising option. In this study, milled ZVI particles (1 g L-1 of particle concentration) were stabilized by environmentally friendly polymer agar agar (>0.5 g L-1), which had a positive impact on the milled ZVI stability. Sedimentation rate was significantly decreased by increasing the suspension viscosity. Column transport experiments were performed for bare and agar agar stabilized milled ZVI particles in commercially available fine grained quartz sand (DORSILIT® Nr.8, Gebrüder Dorfner GmbH Co, Germany) and different porous media collected from brownfields. The experiments were carried out under field relevant injection conditions of 100 m d-1. The maximal travel distance (LT) of less than 10 cm was determined for non-stabilized suspension in fine grained quartz sand, while agar agar (1 g L-1) stabilized milled ZVI suspension revealed LT of 12 m. Similar results were observed for porous media from brownfields showing that mobility of agar agar stabilized particle suspensions was significantly improved compared to bare particles. Based on the mobility data, agar agar stabilized milled zero-valent iron particles could be used for in situ application. Finally, lab-scale batch degradation

  7. Comparison of dosimetry gels prepared by agar and bovine gelatine

    NASA Astrophysics Data System (ADS)

    Sağsöz, M. E.; Korkut, Ö.; Alemdar, N.; Aktaş, S.; Çalı, E. B.; Kantarcı, M.

    2016-04-01

    Gel dosimeters are unique materials capable of showing three dimensional (3D) dose distributions of therapeutic or diagnostic exposures. Fricke gel dosimeters can be considered as chemical dosimeters that rely on a radiation-induced chemical reaction. Dose distribution of Fricke solutions containing Fe+2 ions determines the transformation of acidic, oxygen saturated Fe+2 ions to Fe+3 ions by the ionizing radiation in aqueous solutions. In this study we produced two different types of gel dosimeters using agar and bovine gelatin with similar fabrication methods. We compared the magnetic resonance (MR) T1 imaging responses of these two gel dosimeters to acquire a dose dependency of MR intensities. In conclusion agar gel dosimeters found to be produced easily and more consistent.

  8. Luminescent DNA- and agar-based membranes.

    PubMed

    Leones, R; Fernandes, M; Ferreira, R A S; Cesarino, I; Lima, J F; Carlos, L D; Bermudez, V de Zea; Magon, C J; Donoso, J P; Silva, M M; Pawlicka, A

    2014-09-01

    Luminescent materials containing europium ions are investigated for different optical applications. They can be obtained using bio-macromolecules, which are promising alternatives to synthetic polymers based on the decreasing oil resources. This paper describes studies of the DNA- and Agar-europium triflate luminescent membranes and its potential technological applications are expanded to electroluminescent devices. Polarized optical microscopy demonstrated that the samples are birefringent with submicrometer anisotropy. The X-ray diffraction analysis revealed predominantly amorphous nature of the samples and the atomic force microscopy images showed a roughness of the membranes of 409.0 and 136.1 nm for the samples of DNA10Eu and Agar1.11Eu, respectively. The electron paramagnetic resonance spectra of the DNA(n)Eu membranes with the principal lines at g ≈ 2.0 and g ≈ 4.8 confirmed uniform distribution of rare earth ions in a disordered matrix. Moreover, these strong and narrow resonance lines for the samples of DNA(n)Eu when compared to the Agar(n)Eu suggested a presence of paramagnetic radicals arising from the DNA matrix. The emission spectra suggested that the Eu3+ ions occupy a single local environment in both matrices and the excitation spectra monitored around the Eu emission lines pointed out that the Eu3+ ions in the Agar host were mainly excited via the broad band component rather than by direct intra-4f(6) excitation, whereas the opposite case occurred for the DNA-based sample.

  9. Cluster approach to dilute magnetism

    NASA Astrophysics Data System (ADS)

    Holvorcem, Paulo R. C.; Osório, Roberto

    1988-10-01

    A cluster algebra is developed for the definition of independent correlation functions in the cluster-variation method (CVM) for the spin-1 Ising model. A scheme is then introduced for the study of site-dilute spin- {1}/{2} Ising models by means of the CVM. The procedure regards the site-dilute spin- {1}/{2} model as the spin-1 model with additional constraints due to dilution. The Desjardins-Steinsvoll algortihm is used for the transformation of the CVM equations into a set of differential equations for the independent correlation functions with the inverse temperature as parameter. The evolution of the correlation functions with temperature and the behavior of response functions such as the specific heat and the susceptability are then obtained for any degree of dilution. As an introduction to this scheme, its detailed application is presented here for the simple case of the pair approximation.

  10. Rapid Direct Testing of Susceptibility of Mycobacterium tuberculosis to Isoniazid and Rifampin on Nutrient and Blood Agar in Resource-Starved Settings

    PubMed Central

    Ikram, Aamer; Coban, Ahmet Yilmaz; Martin, Anandi

    2012-01-01

    In this study, we evaluated the performance of blood agar (by macroscopic growth) and nutrient agar (by a microcolony detection method) for drug susceptibility testing of Mycobacterium tuberculosis against rifampin (RIF) and isoniazid (INH), using 67 smear-positive sputum specimens. The direct proportion method on Lowenstein-Jensen (LJ) medium was used as the “gold standard.” Compared with LJ medium, results for both media were in 100% agreement for RIF, while for INH the agreement levels for blood agar and nutrient agar were 98% and 95%, respectively. Within 2 weeks, 100% of specimens yielded results on blood agar, while 96.8% of specimens yielded results on nutrient agar. Our study showed that blood agar and nutrient agar can be used as alternative media for direct susceptibility testing of RIF and INH, especially in resource-poor settings. PMID:22357498

  11. Rapid direct testing of susceptibility of Mycobacterium tuberculosis to isoniazid and rifampin on nutrient and blood agar in resource-starved settings.

    PubMed

    Satti, Luqman; Ikram, Aamer; Coban, Ahmet Yilmaz; Martin, Anandi

    2012-05-01

    In this study, we evaluated the performance of blood agar (by macroscopic growth) and nutrient agar (by a microcolony detection method) for drug susceptibility testing of Mycobacterium tuberculosis against rifampin (RIF) and isoniazid (INH), using 67 smear-positive sputum specimens. The direct proportion method on Lowenstein-Jensen (LJ) medium was used as the "gold standard." Compared with LJ medium, results for both media were in 100% agreement for RIF, while for INH the agreement levels for blood agar and nutrient agar were 98% and 95%, respectively. Within 2 weeks, 100% of specimens yielded results on blood agar, while 96.8% of specimens yielded results on nutrient agar. Our study showed that blood agar and nutrient agar can be used as alternative media for direct susceptibility testing of RIF and INH, especially in resource-poor settings.

  12. Evaluation of Granada agar plate for detection of Streptococcus agalactiae in urine specimens from pregnant women.

    PubMed

    Tamayo, Javier; Gómez-Garcés, José-Luis; Alós, Juan-Ignacio

    2004-08-01

    The Granada agar plate (GAP; Biomedics SL, Madrid, Spain) was evaluated for the detection of group B streptococci (GBS) in urine specimens from pregnant women submitted for testing for asymptomatic bacteriuria and was compared with blood agar (BA [Columbia agar with 5% sheep blood]; bioMérieux, Marcy l'Etoile, France). The GAP detected 103 out of 105 GBS, whereas BA detected only 50. Use of the GAP could be a good method for the detection of GBS in urine specimens from pregnant women. PMID:15297542

  13. Modeling of the Bacillus subtilis Bacterial Biofilm Growing on an Agar Substrate

    PubMed Central

    Wang, Xiaoling; Wang, Guoqing; Hao, Mudong

    2015-01-01

    Bacterial biofilms are organized communities composed of millions of microorganisms that accumulate on almost any kinds of surfaces. In this paper, a biofilm growth model on an agar substrate is developed based on mass conservation principles, Fick's first law, and Monod's kinetic reaction, by considering nutrient diffusion between biofilm and agar substrate. Our results show biofilm growth evolution characteristics such as biofilm thickness, active biomass, and nutrient concentration in the agar substrate. We quantitatively obtain biofilm growth dependence on different parameters. We provide an alternative mathematical method to describe other kinds of biofilm growth such as multiple bacterial species biofilm and also biofilm growth on various complex substrates. PMID:26355542

  14. Modeling of the Bacillus subtilis Bacterial Biofilm Growing on an Agar Substrate.

    PubMed

    Wang, Xiaoling; Wang, Guoqing; Hao, Mudong

    2015-01-01

    Bacterial biofilms are organized communities composed of millions of microorganisms that accumulate on almost any kinds of surfaces. In this paper, a biofilm growth model on an agar substrate is developed based on mass conservation principles, Fick's first law, and Monod's kinetic reaction, by considering nutrient diffusion between biofilm and agar substrate. Our results show biofilm growth evolution characteristics such as biofilm thickness, active biomass, and nutrient concentration in the agar substrate. We quantitatively obtain biofilm growth dependence on different parameters. We provide an alternative mathematical method to describe other kinds of biofilm growth such as multiple bacterial species biofilm and also biofilm growth on various complex substrates.

  15. Serial Dilution Simulation Lab

    ERIC Educational Resources Information Center

    Keler, Cynthia; Balutis, Tabitha; Bergen, Kim; Laudenslager, Bryanna; Rubino, Deanna

    2010-01-01

    Serial dilution is often a difficult concept for students to understand. In this short dry lab exercise, students perform serial dilutions using seed beads. This exercise helps students gain skill at performing dilutions without using reagents, bacterial cultures, or viral cultures, while being able to visualize the process.

  16. Evaluation of a new most-probable-number (MPN) dilution plate method for the enumeration of Escherichia coli in water samples.

    PubMed

    Kodaka, Hidemasa; Saito, Mikako; Matsuoka, Hideaki

    2009-09-01

    The purpose of this study was to evaluate the most-probable-number dilution plate (MPN plate) method developed for the enumeration of Escherichia coil in water samples. Sterilized water was inoculated with E. coli ATCC 11775 to give between 2-1600 MPN/100 ml. The MPN was determined for both the MPN plate and 5-tube methods from the MPN table. The average of the natural logarithm (In) MPN with standard deviations in 95 samples was 4.26 +/- 1.48 by the 5-tube-method and 4.18 +/- 1.45 by the MPN plate method. The correlation coefficient was 0.96. These results were not significantly different according to the paired t-test (p > 0.05).

  17. A hidden pitfall in the preparation of agar media undermines microorganism cultivability.

    PubMed

    Tanaka, Tomohiro; Kawasaki, Kosei; Daimon, Serina; Kitagawa, Wataru; Yamamoto, Kyosuke; Tamaki, Hideyuki; Tanaka, Michiko; Nakatsu, Cindy H; Kamagata, Yoichi

    2014-12-01

    Microbiologists have been using agar growth medium for over 120 years. It revolutionized microbiology in the 1890s when microbiologists were seeking effective methods to isolate microorganisms, which led to the successful cultivation of microorganisms as single clones. But there has been a disparity between total cell counts and cultivable cell counts on plates, often referred to as the "great plate count anomaly," that has long been a phenomenon that still remains unsolved. Here, we report that a common practice microbiologists have employed to prepare agar medium has a hidden pitfall: when phosphate was autoclaved together with agar to prepare solid growth media (PT medium), total colony counts were remarkably lower than those grown on agar plates in which phosphate and agar were separately autoclaved and mixed right before solidification (PS medium). We used a pure culture of Gemmatimonas aurantiaca T-27(T) and three representative sources of environmental samples, soil, sediment, and water, as inocula and compared colony counts between PT and PS agar plates. There were higher numbers of CFU on PS medium than on PT medium using G. aurantiaca or any of the environmental samples. Chemical analysis of PT agar plates suggested that hydrogen peroxide was contributing to growth inhibition. Comparison of 454 pyrosequences of the environmental samples to the isolates revealed that taxa grown on PS medium were more reflective of the original community structure than those grown on PT medium. Moreover, more hitherto-uncultivated microbes grew on PS than on PT medium.

  18. A Hidden Pitfall in the Preparation of Agar Media Undermines Microorganism Cultivability

    PubMed Central

    Tanaka, Tomohiro; Kawasaki, Kosei; Daimon, Serina; Kitagawa, Wataru; Yamamoto, Kyosuke; Tamaki, Hideyuki; Tanaka, Michiko; Nakatsu, Cindy H.

    2014-01-01

    Microbiologists have been using agar growth medium for over 120 years. It revolutionized microbiology in the 1890s when microbiologists were seeking effective methods to isolate microorganisms, which led to the successful cultivation of microorganisms as single clones. But there has been a disparity between total cell counts and cultivable cell counts on plates, often referred to as the “great plate count anomaly,” that has long been a phenomenon that still remains unsolved. Here, we report that a common practice microbiologists have employed to prepare agar medium has a hidden pitfall: when phosphate was autoclaved together with agar to prepare solid growth media (PT medium), total colony counts were remarkably lower than those grown on agar plates in which phosphate and agar were separately autoclaved and mixed right before solidification (PS medium). We used a pure culture of Gemmatimonas aurantiaca T-27T and three representative sources of environmental samples, soil, sediment, and water, as inocula and compared colony counts between PT and PS agar plates. There were higher numbers of CFU on PS medium than on PT medium using G. aurantiaca or any of the environmental samples. Chemical analysis of PT agar plates suggested that hydrogen peroxide was contributing to growth inhibition. Comparison of 454 pyrosequences of the environmental samples to the isolates revealed that taxa grown on PS medium were more reflective of the original community structure than those grown on PT medium. Moreover, more hitherto-uncultivated microbes grew on PS than on PT medium. PMID:25281372

  19. Blood agar validation for susceptibility testing of isoniazid, rifampicin, ethambutol, and streptomycin to Mycobacterium tuberculosis isolates.

    PubMed

    Coban, Ahmet Yilmaz

    2013-01-01

    In recent studies, it was shown that blood agar can be used at least as effectively as Löwenstein-Jensen medium for growing Mycobacterium tuberculosis. It was also shown that susceptibility testing can be performed on blood agar. Additional validation of blood agar was performed on regional M. tuberculosis isolates from Turkey to determine critical concentrations of isoniazid (INH), rifampicin (RIF), ethambutol (ETM), and streptomycin (STR). In the current study, 40 M. tuberculosis clinical isolates were tested. H37Rv, which is susceptible to all antituberculosis agents, ATCC 35822 (INH-resistant), ATCC 35838 (RIF-resistant), ATCC 35837 (ETM-resistant), and ATCC 35820 (STR-resistant) quality control strains were used as control strains. Proportion method on 7H11 agar was considered as gold standard in the study. MIC values of the control strains and clinical isolates were detected on blood and 7H11 agar. Categorical agreements were 100% for each antibiotic, and essential agreements were 100%, 97.5%, 82.5%, and 95% for INH, RIF, ETM, and STR, respectively. According to the data, 0.2 µg/mL for INH, 1 µg/mL for RIF, 4 µg/mL for ETM, and 2 µg/mL for STR were appropriate breakpoint values for susceptibility testing on blood agar. Blood agar may be recommended for use in both developed and developing countries for the susceptibility testing of M. tuberculosis isolates to primary antituberculosis drugs.

  20. Luminescent DNA- and agar-based membranes.

    PubMed

    Leones, R; Fernandes, M; Ferreira, R A S; Cesarino, I; Lima, J F; Carlos, L D; Bermudez, V de Zea; Magon, C J; Donoso, J P; Silva, M M; Pawlicka, A

    2014-09-01

    Luminescent materials containing europium ions are investigated for different optical applications. They can be obtained using bio-macromolecules, which are promising alternatives to synthetic polymers based on the decreasing oil resources. This paper describes studies of the DNA- and Agar-europium triflate luminescent membranes and its potential technological applications are expanded to electroluminescent devices. Polarized optical microscopy demonstrated that the samples are birefringent with submicrometer anisotropy. The X-ray diffraction analysis revealed predominantly amorphous nature of the samples and the atomic force microscopy images showed a roughness of the membranes of 409.0 and 136.1 nm for the samples of DNA10Eu and Agar1.11Eu, respectively. The electron paramagnetic resonance spectra of the DNA(n)Eu membranes with the principal lines at g ≈ 2.0 and g ≈ 4.8 confirmed uniform distribution of rare earth ions in a disordered matrix. Moreover, these strong and narrow resonance lines for the samples of DNA(n)Eu when compared to the Agar(n)Eu suggested a presence of paramagnetic radicals arising from the DNA matrix. The emission spectra suggested that the Eu3+ ions occupy a single local environment in both matrices and the excitation spectra monitored around the Eu emission lines pointed out that the Eu3+ ions in the Agar host were mainly excited via the broad band component rather than by direct intra-4f(6) excitation, whereas the opposite case occurred for the DNA-based sample. PMID:25924317

  1. Determination of low methylmercury concentrations in peat soil samples by isotope dilution GC-ICP-MS using distillation and solvent extraction methods.

    PubMed

    Pietilä, Heidi; Perämäki, Paavo; Piispanen, Juha; Starr, Mike; Nieminen, Tiina; Kantola, Marjatta; Ukonmaanaho, Liisa

    2015-04-01

    Most often, only total mercury concentrations in soil samples are determined in environmental studies. However, the determination of extremely toxic methylmercury (MeHg) in addition to the total mercury is critical to understand the biogeochemistry of mercury in the environment. In this study, N2-assisted distillation and acidic KBr/CuSO4 solvent extraction methods were applied to isolate MeHg from wet peat soil samples collected from boreal forest catchments. Determination of MeHg was performed using a purge and trap GC-ICP-MS technique with a species-specific isotope dilution quantification. Distillation is known to be more prone to artificial MeHg formation compared to solvent extraction which may result in the erroneous MeHg results, especially with samples containing high amounts of inorganic mercury. However, methylation of inorganic mercury during the distillation step had no effect on the reliability of the final MeHg results when natural peat soil samples were distilled. MeHg concentrations determined in peat soil samples after distillation were compared to those determined after the solvent extraction method. MeHg concentrations in peat soil samples varied from 0.8 to 18 μg kg(-1) (dry weight) and the results obtained with the two different methods did not differ significantly (p=0.05). The distillation method with an isotope dilution GC-ICP-MS was shown to be a reliable method for the determination of low MeHg concentrations in unpolluted soil samples. Furthermore, the distillation method is solvent-free and less time-consuming and labor-intensive when compared to the solvent extraction method.

  2. Low density, microcellular, dopable, agar/gelatin foams for pulsed power experiments

    SciTech Connect

    McNamara, W.F.; Aubert, J.H.

    1997-04-01

    Low-density, microcellular foams prepared from the natural polymers agar and gelatin have been developed for pulsed-power physics experiments. Numerous experiments were supported with foams having densities at or below 10 mg/cm{sup 3}. For some of the experiments, the agar/gelatin foam was uniformly doped with metallic elements using soluble salts. Depending on the method of preparation, cell sizes were typically below 10 microns and for one process were below 1.0 micron.

  3. Cross-correlation focus method with an electrostatic sensor array for local particle velocity measurement in dilute gas-solid two-phase flow

    NASA Astrophysics Data System (ADS)

    Wang, Chao; Zhang, Jingyu; Gao, Wenbin; Ding, Hongbing; Wu, Weiping

    2015-11-01

    The gas-solid two-phase flow has been widely applied in the power, chemical and metallurgical industries. It is of great significance in the research of gas-solid two-phase flow to measure particle velocity at different locations in the pipeline. Thus, an electrostatic sensor array comprising eight arc-shaped electrodes was designed. The relationship between the cross-correlation (CC) velocity and the distribution of particle velocity, charge density and electrode spatial sensitivity was analysed. Then the CC sensitivity and its calculation method were proposed. According to the distribution of CC sensitivity, it was found that, between different electrode pairs, it had different focus areas. The CC focus method was proposed for particle velocity measurement at different locations and validated by a belt-style electrostatic induction experiment facility. Finally, the particle velocities at different locations with different flow conditions were measured to research the particle velocity distribution in a dilute horizontal pneumatic conveying pipeline.

  4. Evaluation of Petrifilm™ aerobic count plates as an equivalent alternative to drop plating on R2A agar plates in a biofilm disinfectant efficacy test.

    PubMed

    Fritz, B G; Walker, D K; Goveia, D E; Parker, A E; Goeres, D M

    2015-03-01

    This paper compares Petrifilm™ aerobic count (AC) plates to drop plating on R2A agar plates as an alternative method for biofilm bacteria enumeration after application of a disinfectant. A Pseudomonas aeruginosa biofilm was grown in a Centers for Disease Control and Prevention biofilm reactor (ASTM E2562) and treated with 123 ppm sodium hypochlorite (as free chlorine) according to the Single Tube Method (ASTM E2871). Aliquots from the same dilution tubes were plated on Petrifilm™ AC plates and drop plated on R2A agar plates. The Petrifilm™ AC and R2A plates were incubated for 48 and 24 h, respectively, at 36 ± 1 °C. After nine experimental runs performed by two technicians, the mean difference in biofilm log densities [log biofilm density (LD) = log10(CFU/cm(2))] between the two methods for control coupons, treated coupons, and log reduction (LR) was 0.052 (p = 0.451), -0.102 (p = 0.303), and 0.152 (p = 0.313). Equivalence testing was used to assess equivalence of the two plating methods. The 90 % confidence intervals for the difference in control and treated mean LDs between methods were (-0.065, 0.170) and (-0.270, 0.064), both of which fall within a (-0.5, +0.5) equivalence criterion. The 90 % confidence interval for the mean LR difference (-0.113, 0.420) also falls within this equivalence criterion. Thus, Petrifilm™ AC plates were shown to be statistically equivalent to drop plating on R2A agar for the determination of control LDs, treated LDs, and LR values in an anti-biofilm efficacy test. These are the first published results that establish equivalency to a traditional plate counting technique for biofilms and for a disinfectant assay.

  5. Evaluation of the chromogenic agar chromID C. difficile.

    PubMed

    Eckert, Catherine; Burghoffer, Béatrice; Lalande, Valérie; Barbut, Frederic

    2013-03-01

    Three selective media (chromID C. difficile agar, taurocholate cycloserine cefoxitin agar [TCCA; homemade], and CLO medium) were compared from 406 stool samples of patients suspected of having Clostridium difficile infection. The sensitivities of chromID C. difficile agar at 24 h and 48 h, CLO medium, and TCCA were 74.1%, 87%, 85.2%, and 70.4%, respectively.

  6. Multicenter Investigation of Gepotidacin (GSK2140944) Agar Dilution Quality Control Determinations for Neisseria gonorrhoeae ATCC 49226.

    PubMed

    Jones, Ronald N; Fedler, Kelley A; Scangarella-Oman, Nicole E; Ross, James E; Flamm, Robert K

    2016-07-01

    Gepotidacin, a novel triazaacenaphthylene antibacterial agent, is the first in a new class of type IIA topoisomerase inhibitors with activity against many biothreat and conventional pathogens, including Neisseria gonorrhoeae To assist ongoing clinical studies of gepotidacin to treat gonorrhea, a multilaboratory quality assurance investigation determined the reference organism (N. gonorrhoeae ATCC 49226) quality control MIC range to be 0.25 to 1 μg/ml (88.8% of gepotidacin MIC results at the 0.5 μg/ml mode). PMID:27161642

  7. Microfluidic serial dilution ladder.

    PubMed

    Ahrar, Siavash; Hwang, Michelle; Duncan, Philip N; Hui, Elliot E

    2014-01-01

    Serial dilution is a fundamental procedure that is common to a large number of laboratory protocols. Automation of serial dilution is thus a valuable component for lab-on-a-chip systems. While a handful of different microfluidic strategies for serial dilution have been reported, approaches based on continuous flow mixing inherently consume larger amounts of sample volume and chip real estate. We employ valve-driven circulatory mixing to address these issues and also introduce a novel device structure to store each stage of the dilution process. The dilution strategy is based on sequentially mixing the rungs of a ladder structure. We demonstrate a 7-stage series of 1 : 1 dilutions with R(2) equal to 0.995 in an active device area of 1 cm(2).

  8. Agar-Gel Precipitin Technique in Anthrax Antibody Determinations1

    PubMed Central

    Ray, John G.; Kadull, Paul J.

    1964-01-01

    A modification of the agar-gel precipitation inhibition technique of Thorne and Belton for detecting anthrax antibodies reduces inconsistency of visually determined end points on the same sera observed by different technicians. Determination of the minimal reacting concentrations of the anthrax antigen and antibody reagents, modifications of the visualization apparatus, methods for combining reagents, and length of incubation periods contribute to the ease of the end-point determinations and the uniformity of results. When compared with the previous technique, the modified procedure is less time-consuming while retaining satisfactory reproducibility, simplicity, specificity, and sensitivity. Images FIG. 1 FIG. 2 PMID:14201088

  9. The mathematics of dilution.

    PubMed

    Chatterjee, Barun Kumar

    2014-04-01

    The major objection to homeopathic medicine is that the doses of medicine prescribed in some cases are too dilute for any active ingredient to be present. The medicines would hence be rendered inactive, necessitating novel explanations for the action. A further examination of dilution in the light of the Langmuir equation shows that homeopathic medicines may not be as dilute as a simplistic application of Avogadro's Principle suggests, due to surface effects.

  10. Syneresis and delayed detachment in agar plates

    NASA Astrophysics Data System (ADS)

    Divoux, Thibaut; Mao, Bosi; Snabre, Patrick

    Biogels made of crosslinked polymers such as proteins or polysaccharides behave as porous soft solids and store large amount of solvent. These gels undergo spontaneous aging, called syneresis that consists in the shrinkage of the gel matrix and the progressive expulsion of the solvent. As a result, a biogel originally casted in a container often lose contact with the container sidewalls, and the detachment time is a priori difficult to anticipate since it may occur over variable time spans (from hours to days). Here we report on the syneresis phenomena in agar plates that consist in Petri dishes filled with a gel mainly composed of agar. Direct observations and speckle pattern correlation analysis allow us to rationalize the delayed detachment of the gel from the sidewall of the Petri dish. The detachment time $t^*$ is surprisingly not controlled by the mass loss as one would intuitively expect. Instead, $t^*$ is strongly correlated to the gel minimum thickness $e_{min}$ measured along the sidewall of the plate, and increases as a robust function of $e_{min}$ independently of the prior mass-loss history. Time-resolved correlation spectroscopy atypically applied to such weakly diffusive media gives access to the local thinning rate of the gel. This technique also allows us to detect the gel micro-displacements that are triggered by the water evaporation prior to the detachment, and even to anticipate the latter from a few hours. Our work provides observables to predict the detachment time of agar gels in dishes, and highlights the relevance of speckle pattern correlation analysis for the quantitative investigation of the syneresis dynamics in biopolymer gels.

  11. Syneresis and delayed detachment in agar plates.

    PubMed

    Divoux, Thibaut; Mao, Bosi; Snabre, Patrick

    2015-05-14

    Biogels made of crosslinked polymers such as proteins or polysaccharides behave as porous soft solids and store large amounts of solvent. These gels undergo spontaneous aging, called syneresis, which consists of the shrinkage of the gel matrix and the progressive expulsion of solvent. As a result, a biogel originally casted in a container often loses contact with the container sidewalls, and the detachment time is difficult to anticipate a priori, since it may occur over variable time spans (from hours to days). Here we report on syneresis phenomena in agar plates, which consist of Petri dishes filled with a gel mainly composed of agar. Direct observations and speckle pattern correlation analysis allow us to rationalize the delayed detachment of the gel from the sidewall of the Petri dish. The detachment time t* is surprisingly not controlled by the mass loss as one would intuitively expect. Instead, t* is strongly correlated to the gel minimum thickness emin measured along the sidewall of the plate, and increases as a robust function of emin, independently of the prior mass-loss history. Time-resolved correlation spectroscopy atypically applied to such weakly diffusive media gives access to the local thinning rate of the gel. This technique also allows us to detect the gel micro-displacements that are triggered by water evaporation prior to the detachment, and even to anticipate the latter from a few hours. Our work provides observables to predict the detachment time of agar gels in dishes, and highlights the relevance of speckle pattern correlation analysis for the quantitative investigation of the syneresis dynamics in biopolymer gels.

  12. Syneresis and delayed detachment in agar plates.

    PubMed

    Divoux, Thibaut; Mao, Bosi; Snabre, Patrick

    2015-05-14

    Biogels made of crosslinked polymers such as proteins or polysaccharides behave as porous soft solids and store large amounts of solvent. These gels undergo spontaneous aging, called syneresis, which consists of the shrinkage of the gel matrix and the progressive expulsion of solvent. As a result, a biogel originally casted in a container often loses contact with the container sidewalls, and the detachment time is difficult to anticipate a priori, since it may occur over variable time spans (from hours to days). Here we report on syneresis phenomena in agar plates, which consist of Petri dishes filled with a gel mainly composed of agar. Direct observations and speckle pattern correlation analysis allow us to rationalize the delayed detachment of the gel from the sidewall of the Petri dish. The detachment time t* is surprisingly not controlled by the mass loss as one would intuitively expect. Instead, t* is strongly correlated to the gel minimum thickness emin measured along the sidewall of the plate, and increases as a robust function of emin, independently of the prior mass-loss history. Time-resolved correlation spectroscopy atypically applied to such weakly diffusive media gives access to the local thinning rate of the gel. This technique also allows us to detect the gel micro-displacements that are triggered by water evaporation prior to the detachment, and even to anticipate the latter from a few hours. Our work provides observables to predict the detachment time of agar gels in dishes, and highlights the relevance of speckle pattern correlation analysis for the quantitative investigation of the syneresis dynamics in biopolymer gels. PMID:25812667

  13. Characteristic features and dye degrading capability of agar-agar gel immobilized manganese peroxidase.

    PubMed

    Bilal, Muhammad; Asgher, Muhammad; Shahid, Muhammad; Bhatti, Haq Nawaz

    2016-05-01

    Immobilization of enzymes has been regarded as an efficient approach to develop biocatalyst with improved activity and stability characteristics under reaction conditions. In the present study, purified manganese peroxidase (MnP) from Ganoderma lucidum IBL-05 was immobilized in agar-agar support using entrapment technique. Maximum immobilization yield was accomplished at 4.0% agar-agar gel. The immobilized MnP exhibited better resistance to changes in pH and temperature than the free enzyme, with optimal conditions being pH 6.0 and 50 °C. The kinetic parameters Km and Kcat/Km for free and entrapped MnP were calculated to be 65.6 mM and 6.99 M(-1) s(-1), and 82 mM and 8.15 M(-1) s(-1), respectively. Thermo-stability was significantly improved after immobilization. After 120 h, the insolubilized MnP retained its activity up to 71.9% and 60.3% at 30 °C and 40 °C, respectively. It showed activity until 10th cycle and retained 74.3% residual activity after 3th cycle. The effects of H2O2, ionic strength and potential inhibitors on activity of free and immobilized enzyme were investigated. Moreover, the decolorization of three structurally different dyes was monitored in order to assess the degrading capability of the entrapped MnP. The decolorization efficiencies for all the tested dyes were 78.6-84.7% after 12h. The studies concluded that the toxicity of dyes aqueous solutions was significantly reduced after treatment. The remarkable catalytic, thermo-stability and re-cycling features of the agar-agar immobilized MnP display a high potential for biotechnological applications. PMID:26854887

  14. Validation of an isotope dilution gas chromatography-mass spectrometry method for combined analysis of oxysterols and oxyphytosterols in serum samples.

    PubMed

    Schött, Hans-Frieder; Lütjohann, Dieter

    2015-07-01

    We describe the validation of a method for the analysis of oxysterols, i.e. oxycholesterols and oxyphytosterols, in human serum using gas chromatography-mass spectrometry selected ion monitoring (GC-MS-SIM). Concentrations of 7α- and 7β-hydroxy-, and 7oxo-cholesterol, -campesterol, and -sitosterol as well as 4β-hydroxycholesterol and side-chain oxygenated 24S-, 25-, and 27-hydroxycholesterol were determined by isotope dilution methodology. After saponification at room temperature the oxysterols were extracted, separated from their substrates, cholesterol, campesterol, and sitosterol, by solid phase extraction, and subsequently derivatised to their corresponding trimethylsilyl-ethers prior to GC-MS-SIM. In order to prevent artificial autoxidation butylated hydroxytoluene and ethylenediaminetetraacetic acid were added. The validation of the method was performed according to the International Conference on Harmonisation guidance, including limits of detection and quantification, ranges, recovery and precision. Due to improved instrumental settings and work-up procedure, limits of detection and quantification ranged between 8.0-202.0pg/mL and 28.0-674pg/mL, respectively. Recovery data in five calibration points varied between 91.9% and 116.8% and in serum samples between 93.1% and 118.1%. The mean coefficient of variation (CV) for the recovery of all compounds was <10%. Well satisfying CVs for within-day precision (2.1-10.8%) and for between-day precision (2.3-12.1%) were obtained. More than 20 samples could be processed in a single routine day and test series of about 300 samples can be realised without impairment of the validation parameters during a sequence. Comparison of oxysterol and oxyphytosterol content in serum and plasma revealed no difference. A fully validated isotope dilution methodology for the quantification of oxycholesterols and oxyphytosterols from human serum or plasma is presented.

  15. Validation of an isotope dilution gas chromatography-mass spectrometry method for combined analysis of oxysterols and oxyphytosterols in serum samples.

    PubMed

    Schött, Hans-Frieder; Lütjohann, Dieter

    2015-07-01

    We describe the validation of a method for the analysis of oxysterols, i.e. oxycholesterols and oxyphytosterols, in human serum using gas chromatography-mass spectrometry selected ion monitoring (GC-MS-SIM). Concentrations of 7α- and 7β-hydroxy-, and 7oxo-cholesterol, -campesterol, and -sitosterol as well as 4β-hydroxycholesterol and side-chain oxygenated 24S-, 25-, and 27-hydroxycholesterol were determined by isotope dilution methodology. After saponification at room temperature the oxysterols were extracted, separated from their substrates, cholesterol, campesterol, and sitosterol, by solid phase extraction, and subsequently derivatised to their corresponding trimethylsilyl-ethers prior to GC-MS-SIM. In order to prevent artificial autoxidation butylated hydroxytoluene and ethylenediaminetetraacetic acid were added. The validation of the method was performed according to the International Conference on Harmonisation guidance, including limits of detection and quantification, ranges, recovery and precision. Due to improved instrumental settings and work-up procedure, limits of detection and quantification ranged between 8.0-202.0pg/mL and 28.0-674pg/mL, respectively. Recovery data in five calibration points varied between 91.9% and 116.8% and in serum samples between 93.1% and 118.1%. The mean coefficient of variation (CV) for the recovery of all compounds was <10%. Well satisfying CVs for within-day precision (2.1-10.8%) and for between-day precision (2.3-12.1%) were obtained. More than 20 samples could be processed in a single routine day and test series of about 300 samples can be realised without impairment of the validation parameters during a sequence. Comparison of oxysterol and oxyphytosterol content in serum and plasma revealed no difference. A fully validated isotope dilution methodology for the quantification of oxycholesterols and oxyphytosterols from human serum or plasma is presented. PMID:25701095

  16. Determination of single and repeated red cell volumes by the indicator dilution method using carbon monoxide as the indicator

    SciTech Connect

    Fukui, M.; Shigemi, K. )

    1989-11-01

    The use of radioactive isotopes limits clinical applications of blood volume measurement in the ICU. We measured red cell volumes with carbon monoxide-labeled RBC in six dogs and five human volunteers. The measured values obtained on the dogs were compared with the simultaneous measurements with the {sup 51}Cr method; the ratio of the carbon monoxide to {sup 51}Cr values ranged from 0.86 to 1.17, and the mean ratio was 1.0 +/- 0.1 (SD), r = .93. We infer from these results that the carbon monoxide method has several advantages over the {sup 51}Cr method: (a) the short labeling time (about 1 min), (b) rapidly decreasing background levels of carbon monoxide with FIO2 1.0, and (c) repeatability at intervals of several hours.

  17. Biological treatment of textile dyes by agar-agar immobilized consortium in a packed bed reactor.

    PubMed

    Patel, Yogesh; Gupte, Akshaya

    2015-03-01

    The decolorization of Acid Maroon V was investigated using bacterial consortium EDPA containing Enterobacter dissolvens AGYP1 and Pseudomonas aeruginosa AGYP2 immobilized in different entrapment matrices. The consortium displayed 96% removal of dye (100 mg/l) within 6 h when immobilized in agar-agar. Under optimum concentrations of agar-agar (3.0% w/v) and cell biomass (0.9 g% w/v), the consortium displayed decolorization for 18 successive batches of Acid Maroon V and also decolorized 14 other different textile dyes. A packed bed reactor under batch mode showed 89% decolorization of dye after 56 repetitive cycles. Under continuous flow mode, maximum color removal was achieved with bed length of 36 cm, hydraulic retention time of 2.66 h, and dye concentration of 100 mg/l. Additionally, the reactor decolorized relatively higher concentrations (100-2000 mg/l) of dye. The synthetic dye wastewater containing five textile dyes was decolorized 92% with 62% COD reduction using an immobilized consortium.

  18. In Silico Calculation of Infinite Dilution Activity Coefficients of Molecular Solutes in Ionic Liquids: Critical Review of Current Methods and New Models Based on Three Machine Learning Algorithms.

    PubMed

    Paduszyński, Kamil

    2016-08-22

    The aim of the paper is to address all the disadvantages of currently available models for calculating infinite dilution activity coefficients (γ(∞)) of molecular solutes in ionic liquids (ILs)-a relevant property from the point of view of many applications of ILs, particularly in separations. Three new models are proposed, each of them based on distinct machine learning algorithm: stepwise multiple linear regression (SWMLR), feed-forward artificial neural network (FFANN), and least-squares support vector machine (LSSVM). The models were established based on the most comprehensive γ(∞) data bank reported so far (>34 000 data points for 188 ILs and 128 solutes). Following the paper published previously [J. Chem. Inf. Model 2014, 54, 1311-1324], the ILs were treated in terms of group contributions, whereas the Abraham solvation parameters were used to quantify an impact of solute structure. Temperature is also included in the input data of the models so that they can be utilized to obtain temperature-dependent data and thus related thermodynamic functions. Both internal and external validation techniques were applied to assess the statistical significance and explanatory power of the final correlations. A comparative study of the overall performance of the investigated SWMLR/FFANN/LSSVM approaches is presented in terms of root-mean-square error and average absolute relative deviation between calculated and experimental γ(∞), evaluated for different families of ILs and solutes, as well as between calculated and experimental infinite dilution selectivity for separation problems benzene from n-hexane and thiophene from n-heptane. LSSVM is shown to be a method with the lowest values of both training and generalization errors. It is finally demonstrated that the established models exhibit an improved accuracy compared to the state-of-the-art model, namely, temperature-dependent group contribution linear solvation energy relationship, published in 2011 [J. Chem

  19. In Silico Calculation of Infinite Dilution Activity Coefficients of Molecular Solutes in Ionic Liquids: Critical Review of Current Methods and New Models Based on Three Machine Learning Algorithms.

    PubMed

    Paduszyński, Kamil

    2016-08-22

    The aim of the paper is to address all the disadvantages of currently available models for calculating infinite dilution activity coefficients (γ(∞)) of molecular solutes in ionic liquids (ILs)-a relevant property from the point of view of many applications of ILs, particularly in separations. Three new models are proposed, each of them based on distinct machine learning algorithm: stepwise multiple linear regression (SWMLR), feed-forward artificial neural network (FFANN), and least-squares support vector machine (LSSVM). The models were established based on the most comprehensive γ(∞) data bank reported so far (>34 000 data points for 188 ILs and 128 solutes). Following the paper published previously [J. Chem. Inf. Model 2014, 54, 1311-1324], the ILs were treated in terms of group contributions, whereas the Abraham solvation parameters were used to quantify an impact of solute structure. Temperature is also included in the input data of the models so that they can be utilized to obtain temperature-dependent data and thus related thermodynamic functions. Both internal and external validation techniques were applied to assess the statistical significance and explanatory power of the final correlations. A comparative study of the overall performance of the investigated SWMLR/FFANN/LSSVM approaches is presented in terms of root-mean-square error and average absolute relative deviation between calculated and experimental γ(∞), evaluated for different families of ILs and solutes, as well as between calculated and experimental infinite dilution selectivity for separation problems benzene from n-hexane and thiophene from n-heptane. LSSVM is shown to be a method with the lowest values of both training and generalization errors. It is finally demonstrated that the established models exhibit an improved accuracy compared to the state-of-the-art model, namely, temperature-dependent group contribution linear solvation energy relationship, published in 2011 [J. Chem

  20. Comparison of Fecal Coliform Agar and Violet Red Bile Lactose Agar for Fecal Coliform Enumeration in Foods

    PubMed Central

    Leclercq, A.; Wanegue, C.; Baylac, P.

    2002-01-01

    A 24-h direct plating method for fecal coliform enumeration with a resuscitation step (preincubation for 2 h at 37 ± 1°C and transfer to 44 ± 1°C for 22 h) using fecal coliform agar (FCA) was compared with the 24-h standardized violet red bile lactose agar (VRBL) method. FCA and VRBL have equivalent specificities and sensitivities, except for lactose-positive non-fecal coliforms such as Hafnia alvei, which could form typical colonies on FCA and VRBL. Recovery of cold-stressed Escherichia coli in mashed potatoes on FCA was about 1 log unit lower than that with VRBL. When the FCA method was compared with standard VRBL for enumeration of fecal coliforms, based on counting carried out on 170 different food samples, results were not significantly different (P > 0.05). Based on 203 typical identified colonies selected as found on VRBL and FCA, the latter medium appears to allow the enumeration of more true fecal coliforms and has higher performance in certain ways (specificity, sensitivity, and negative and positive predictive values) than VRBL. Most colonies clearly identified on both media were E. coli and H. alvei, a non-fecal coliform. Therefore, the replacement of fecal coliform enumeration by E. coli enumeration to estimate food sanitary quality should be recommended. PMID:11916678

  1. Recovery of spores of Clostridium botulinum in yeast extract agar and pork infusion agar after heat treatment.

    PubMed

    Odlaug, T E; Pflug, I J

    1977-10-01

    Yeast extract agar, pork infusion agar, and modifications of these media were used to recover heated Clostridium botulinum spores. The D- and z-values were determined. Two type A strains and one type B strain of C. botulinum were studied. In all cases the D-values were largest when the spores were recovered in yeast extract agar, compared to the D-values for spores recovered in pork infusion agar. The z-values for strains 62A and A16037 were largest when the spores were recovered in pork infusion agar. The addition of sodium bicarbonate and sodium thioglycolate to pork infusion agar resulted in D-values for C. botulinum 62A spores similar to those for the same spores recovered in yeast extract agar. The results suggest that sodium bicarbonate and sodium thioglycolate should be added to recovery media for heated C. botulinum spores to obtain maximum plate counts. PMID:335970

  2. Direct and selective flow-injection method for the simultaneous spectrophotometric determination of calcium and magnesium in red and white wines using online dilution based on "Zone Sampling".

    PubMed

    Themelis, D G; Tzanavaras, P D; Trellopoulos, A V; Sofoniou, M C

    2001-11-01

    The present work reports a selective and simple flow injection method for the direct and simultaneous determination of calcium and magnesium ions in red, rose, and white wines. Both ions react with methylthymol blue (MTB) at a strongly basic medium to form colored complexes that are monitored spectrophotometrically (lambda(max) = 610 nm). The simultaneous determination is achieved by online masking of magnesium by 8-hydroxyquinoline (8-HQ). Incorporating an online dilution mode based on the "zone sampling" technique in the FI system, the determination of both analytes was achieved without any pretreatment of the samples, in the range 0-350 mg L(-1) and 0-200 mg L(-1) for Ca(II) and Mg(II), respectively. The 3 sigma detection limits were quite satisfactory (2.1 and 1.8 mg L(-1) for Ca(II) and Mg(II) respectively), and the precision was 1.2% (at a mixture of 100.0 mg L(-1) Ca(II) + 100.0 mg L(-1) Mg(II), n = 12). A detailed study of interferences proved that the proposed method is highly selective. The application of the method to the direct analysis of red, rose, and white wines yielded excellent results compared with those obtained by using FAAS as a reference method (e(r) < 2.8%).

  3. Improving agar electrospinnability with choline-based deep eutectic solvents.

    PubMed

    Sousa, Ana M M; Souza, Hiléia K S; Uknalis, Joseph; Liu, Shih-Chuan; Gonçalves, Maria P; Liu, LinShu

    2015-09-01

    Very recently our group has produced novel agar-based fibers by an electrospinning technique using water as solvent and polyvinyl alcohol (PVA) as co-blending polymer. Here, we tested the deep eutectic solvent (DES), (2-hydroxyethyl)trimethylammonium chloride/urea prepared at 1:2 molar ratio, as an alternative solvent medium for agar electrospinning. The electrospun materials were collected with an ethanol bath adapted to a previous electrospinning set-up. One weight percent agar-in-DES showed improved viscoelasticity and hence, spinnability, when compared to 1 wt% agar-in-water and pure agar nanofibers were successfully electrospun if working above the temperature of sol-gel transition (∼80 °C). By changing the solvent medium we decreased the PVA concentration (5 wt% starting solution) and successfully produced composite fibers with high agar contents (50/50 agar/PVA). Best composite fibers were formed with the 50/50 and 30/70 agar/PVA solutions. These fibers were mechanically resistant, showed tailorable surface roughness and diverse size distributions, with most of the diameters falling in the sub-micron range. Both nano and micro forms of agar fibers (used separately or combined) may have potential for the design of new and highly functional agar-based materials. PMID:26116384

  4. Contribution to diagnostics/prognostics of tuberculosis in children. I. New methods of assaying zinc and simultaneously copper and zinc in diluted sera by flame atomic-absorption spectrometry.

    PubMed

    Luterotti, Svjetlana; Kordić, Tončica Vukman; Dodig, Slavica

    2015-09-01

    In an attempt to provide a reliable status of metal ions in children, new methods of analysis of children's sera are proposed. New flame atomic-absorption spectrometric (FAAS) methods are simple, cost- and time-effective and, above all, labor-, reagent- and sample-saving. Two methods were suggested: method A for simultaneous determination of Cu and Zn from 5-fold diluted sera, and method B, for assaying zinc alone in 10-fold diluted samples. Both methods are based on a single-step sample pretreatment (deproteinization with 3 mol dm-3 HCl). Method A uses a single-step calibration with a mixed standard. The main advantage of method B is an additional reduction in sample consumption. Both methods were fully validated against reference methods. Accuracy, sensitivity and precision have proven them to be comparable to the reference methods in terms of analytical performance, and applicable to analyses of children's sera.

  5. 'Dilute-and-shoot' RSLC-MS-MS method for fast detection of nerve and vesicant chemical warfare agent metabolites in urine.

    PubMed

    Rodin, Igor; Braun, Arcady; Stavrianidi, Andrey; Baygildiev, Timur; Shpigun, Oleg; Oreshkin, Dmitry; Rybalchenko, Igor

    2015-01-01

    A sensitive screening method based on fast liquid chromatography tandem mass-spectrometry (RSLC-MS-MS) has shown the feasibility of separation and detection of low concentration β-lyase metabolites of sulfur mustard and of nerve agent phosphonic acids in urine. The analysis of these compounds is of interest because they are specific metabolites of the chemical warfare agents (CWAs), sulfur mustard (HD), sarin (GB), soman (GD), VX and Russian VX (RVX). The 'dilute-and-shoot' RSLC-MS-MS method provides a sensitive and direct approach for determining CWA exposure in non-extracted non-derivatized samples from urine. Chromatographic separation of the metabolites was achieved using a reverse phase column with gradient mobile phases consisting of 0.5% formic acid in water and acetonitrile. Identification and quantification of species were achieved using electrospray ionization-tandem mass-spectrometry monitoring two precursor-to-product ion transitions for each compound. The method demonstrates linearity over at least two orders of magnitude and had detection limits of 0.5 ng/mL in urine.

  6. 'Dilute-and-shoot' RSLC-MS-MS method for fast detection of nerve and vesicant chemical warfare agent metabolites in urine.

    PubMed

    Rodin, Igor; Braun, Arcady; Stavrianidi, Andrey; Baygildiev, Timur; Shpigun, Oleg; Oreshkin, Dmitry; Rybalchenko, Igor

    2015-01-01

    A sensitive screening method based on fast liquid chromatography tandem mass-spectrometry (RSLC-MS-MS) has shown the feasibility of separation and detection of low concentration β-lyase metabolites of sulfur mustard and of nerve agent phosphonic acids in urine. The analysis of these compounds is of interest because they are specific metabolites of the chemical warfare agents (CWAs), sulfur mustard (HD), sarin (GB), soman (GD), VX and Russian VX (RVX). The 'dilute-and-shoot' RSLC-MS-MS method provides a sensitive and direct approach for determining CWA exposure in non-extracted non-derivatized samples from urine. Chromatographic separation of the metabolites was achieved using a reverse phase column with gradient mobile phases consisting of 0.5% formic acid in water and acetonitrile. Identification and quantification of species were achieved using electrospray ionization-tandem mass-spectrometry monitoring two precursor-to-product ion transitions for each compound. The method demonstrates linearity over at least two orders of magnitude and had detection limits of 0.5 ng/mL in urine. PMID:25326204

  7. A Simplified Method for Quantifying Sulfur Mustard Adducts to Blood Proteins by Ultra-High Pressure Liquid Chromatography-Isotope Dilution Tandem Mass Spectrometry

    PubMed Central

    Pantazides, Brooke G.; Crow, Brian S.; Garton, Joshua W.; Quiñones-González, Jennifer A.; Blake, Thomas A.; Thomas, Jerry D.; Johnson, Rudolph C.

    2016-01-01

    Sulfur mustard binds to reactive cysteine residues, forming a stable sulfur-hydroxyethylthioethyl [S-HETE] adduct that can be used as a long-term biomarker of sulfur mustard exposure in humans. The digestion of sulfur mustard-exposed blood samples with proteinase K following total protein precipitation with acetone produces the tripeptide biomarker [S-HETE]-Cys-Pro-Phe. The adducted tripeptide is purified by solid phase extraction, separated by ultra-high pressure liquid chromatography, and detected by isotope dilution tandem mass spectrometry. This approach was thoroughly validated and characterized in our laboratory. The average interday relative standard deviation was ≤ 9.49%, and the range of accuracy was between 96.1-109% over a concentration range of 3.00 to 250. ng/mL with a calculated limit of detection of 1.74 ng/mL. A full 96-well plate can be processed and analyzed in 8 h which is five times faster than our previous 96-well plate method and only requires 50 µL of serum, plasma, or whole blood. Extensive ruggedness and stability studies and matrix comparisons were conducted to create a robust, easily transferrable method. As a result, a simple and high-throughput method has been developed and validated for the quantitation of sulfur mustard blood protein adducts in low volume blood specimens which should be readily adaptable for quantifying human exposures to other alkylating agents. PMID:25622494

  8. Agar Sediment Test for Assessing the Suitability of Organic Waste Streams for Recovering Nutrients by the Aquatic Worm Lumbriculus variegatus.

    PubMed

    Laarhoven, Bob; Elissen, H J H; Temmink, H; Buisman, C J N

    2016-01-01

    An agar sediment test was developed to evaluate the suitability of organic waste streams from the food industry for recovering nutrients by the aquatic worm Lumbriculus variegatus (Lv). The effects of agar gel, sand, and food quantities in the sediment test on worm growth, reproduction, and water quality were studied. Agar gel addition ameliorated growth conditions by reducing food hydrolysis and altering sediment structure. Best results for combined reproduction and growth were obtained with 0.6% agar-gel (20 ml), 10 g. fine sand, 40 g. coarse sand, and 105 mg fish food (Tetramin). With agar gel, ingestion and growth is more the result of addition of food in its original quality. Final tests with secondary potato starch sludge and wheat bran demonstrated that this test is appropriate for the comparison of solid feedstuffs and suspended organic waste streams. This test method is expected to be suitable for organic waste studies using other sediment dwelling invertebrates.

  9. Agar Sediment Test for Assessing the Suitability of Organic Waste Streams for Recovering Nutrients by the Aquatic Worm Lumbriculus variegatus

    PubMed Central

    Laarhoven, Bob; Elissen, H. J. H.; Temmink, H.; Buisman, C. J. N.

    2016-01-01

    An agar sediment test was developed to evaluate the suitability of organic waste streams from the food industry for recovering nutrients by the aquatic worm Lumbriculus variegatus (Lv). The effects of agar gel, sand, and food quantities in the sediment test on worm growth, reproduction, and water quality were studied. Agar gel addition ameliorated growth conditions by reducing food hydrolysis and altering sediment structure. Best results for combined reproduction and growth were obtained with 0.6% agar-gel (20 ml), 10 g. fine sand, 40 g. coarse sand, and 105 mg fish food (Tetramin). With agar gel, ingestion and growth is more the result of addition of food in its original quality. Final tests with secondary potato starch sludge and wheat bran demonstrated that this test is appropriate for the comparison of solid feedstuffs and suspended organic waste streams. This test method is expected to be suitable for organic waste studies using other sediment dwelling invertebrates. PMID:26937632

  10. Agar Sediment Test for Assessing the Suitability of Organic Waste Streams for Recovering Nutrients by the Aquatic Worm Lumbriculus variegatus.

    PubMed

    Laarhoven, Bob; Elissen, H J H; Temmink, H; Buisman, C J N

    2016-01-01

    An agar sediment test was developed to evaluate the suitability of organic waste streams from the food industry for recovering nutrients by the aquatic worm Lumbriculus variegatus (Lv). The effects of agar gel, sand, and food quantities in the sediment test on worm growth, reproduction, and water quality were studied. Agar gel addition ameliorated growth conditions by reducing food hydrolysis and altering sediment structure. Best results for combined reproduction and growth were obtained with 0.6% agar-gel (20 ml), 10 g. fine sand, 40 g. coarse sand, and 105 mg fish food (Tetramin). With agar gel, ingestion and growth is more the result of addition of food in its original quality. Final tests with secondary potato starch sludge and wheat bran demonstrated that this test is appropriate for the comparison of solid feedstuffs and suspended organic waste streams. This test method is expected to be suitable for organic waste studies using other sediment dwelling invertebrates. PMID:26937632

  11. Performance of CHROMAGAR candida and BIGGY agar for identification of yeast species

    PubMed Central

    Yücesoy, Mine; Marol, Serhat

    2003-01-01

    Background The importance of identifying the pathogenic fungi rapidly has encouraged the development of differential media for the presumptive identification of yeasts. In this study two differential media, CHROMagar Candida and bismuth sulphite glucose glycine yeast agar, were evaluated for the presumptive identification of yeast species. Methods A total number of 270 yeast strains including 169 Candida albicans, 33 C. tropicalis, 24 C. glabrata, 18 C. parapsilosis, 12 C. krusei, 5 Trichosporon spp., 4 C. kefyr, 2 C. lusitaniae, 1 Saccharomyces cerevisiae and 1 Geotrichum candidum were included. The strains were first identified by germ tube test, morphological characteristics on cornmeal tween 80 agar and Vitek 32 and API 20 C AUX systems. In parallel, they were also streaked onto CHROMagar Candida and bismuth sulphite glucose glycine yeast agar plates. The results were read according to the color, morphology of the colonies and the existance of halo around them after 48 hours of incubation at 37°C. Results The sensitivity and specificity values for C. albicans strains were found to be 99.4, 100% for CHROMagar Candida and 87.0, 75.2% for BiGGY agar, respectively. The sensitivity of CHROMagar Candida to identify C. tropicalis, C. glabrata and C. krusei ranged between 90.9 and 100% while the specificity was 100%. The sensitivity rates for BiGGY agar were 66.6 and 100% while the specificity values were found to be 95.4 and 100% for C. tropicalis and C. krusei, respectively. Conclusions It can be concluded that the use of CHROMagar Candida is an easy and reliable method for the presumptive identification of most commonly isolated Candida species especially C. albicans, C. tropicalis and C. krusei. The lower sensitivity and specificity of BiGGY agar to identify commonly isolated Candida species potentially limits the clinical usefulness of this agar. PMID:14613587

  12. An optimized method for the accurate determination of patulin in apple products by isotope dilution-liquid chromatography/mass spectrometry.

    PubMed

    Seo, Miyeong; Kim, Byungjoo; Baek, Song-Yee

    2015-07-01

    Patulin, a mycotoxin produced by several molds in fruits, has been frequently detected in apple products. Therefore, regulatory bodies have established recommended maximum permitted patulin concentrations for each type of apple product. Although several analytical methods have been adopted to determine patulin in food, quality control of patulin analysis is not easy, as reliable certified reference materials (CRMs) are not available. In this study, as a part of a project for developing CRMs for patulin analysis, we developed isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS) as a higher-order reference method for the accurate value-assignment of CRMs. (13)C7-patulin was used as internal standard. Samples were extracted with ethyl acetate to improve recovery. For further sample cleanup with solid-phase extraction (SPE), the HLB SPE cartridge was chosen after comparing with several other types of SPE cartridges. High-performance liquid chromatography was performed on a multimode column for proper retention and separation of highly polar and water-soluble patulin from sample interferences. Sample extracts were analyzed by LC/MS/MS with electrospray ionization in negative ion mode with selected reaction monitoring of patulin and (13)C7-patulin at m/z 153→m/z 109 and m/z 160→m/z 115, respectively. The validity of the method was tested by measuring gravimetrically fortified samples of various apple products. In addition, the repeatability and the reproducibility of the method were tested to evaluate the performance of the method. The method was shown to provide accurate measurements in the 3-40 μg/kg range with a relative expanded uncertainty of around 1%.

  13. Comparison of different extraction methods for simultaneous determination of B complex vitamins in nutritional yeast using LC/MS-TOF and stable isotope dilution assay.

    PubMed

    Hälvin, Kristel; Paalme, Toomas; Nisamedtinov, Ildar

    2013-02-01

    The application of LC/MS-TOF method combined with stable isotope dilution assay was studied for determination of thiamine, riboflavin, nicotinamide, nicotinic acid, pantothenic acid, pyridoxal, and pyridoxine in food. Nutritional yeast powder was used as a model food matrix. Acid extraction was compared with various enzymatic treatments in ammonium formate buffer to find a suitable method for the conversion of more complex vitamers into the same forms as the used isotope-labeled internal standards. The enzyme preparations α-amylase, takadiastase, β-glucosidase, and acid phosphatase were all able to liberate thiamine and riboflavin. The diastatic enzyme preparations α-amylase and takadiastase also expressed proteolytic side activities resulting in the formation of small peptides which interfered with the mass spectra of thiamine and riboflavin. Liberation of nicotinamide and pantothenic acid from NAD(+) and CoA, respectively, could not be achieved with any of the studied enzyme preparations. Hydrochloric acid extraction at 121 °C for 30 min was found to be destructive to pantothenic acid, but increased the liberation of pyridoxal.

  14. Simplified method for the determination of Ru, Pd, Re, Os, Ir and Pt in chromitites and other geological materials by isotope dilution ICP-MS and acid digestion.

    PubMed

    Meisel, T; Moser, J; Fellner, N; Wegscheider, W; Schoenberg, R

    2001-03-01

    A method for the determination of low Ru, Pd, Re, Os, Ir and Pt abundances in geological reference materials by isotope dilution inductively coupled plasma mass spectrometry (ICP-MS) after acid digestion in a high pressure asher (HPA-S) is presented. The digestion technique is similar to that using Carius tubes but easier to handle and reaches higher temperatures. Osmium can be determined as OsO4 with ICP-MS directly after digestion through a sparging technique. The remaining elements are preconcentrated by means of anion column chromatography. The resin is digested directly without elution leading to high yields but this causes problems if Zr is present at higher levels in the silicate rich materials. The analytical results for international platinum group element (PGE) reference materials, chromitite CHR-Bkg, basalt TDB-1 and gabbro WGB-1, are presented and compared with literature data, demonstrating the validity of the described method. Although higher in concentration, PGEs determined for reference material WGB-1 were worse than for TDB-1 indicating a more inhomogeneous distribution of the platinum group mineral phases. The low PGE abundance chromitite standard, CHR-Bkg, is likely to be homogeneous for Ru, Re, Os and Ir and is recommended as a reference material for the study of chromitites. Detection limits (3s x total procedure blank) range from 0.012 ng (Re and Os) to 0.77 ng (Pt), which could be further improved by applying higher quality acids.

  15. Simplified method for the determination of Ru, Pd, Re, Os, Ir and Pt in chromitites and other geological materials by isotope dilution ICP-MS and acid digestion.

    PubMed

    Meisel, T; Moser, J; Fellner, N; Wegscheider, W; Schoenberg, R

    2001-03-01

    A method for the determination of low Ru, Pd, Re, Os, Ir and Pt abundances in geological reference materials by isotope dilution inductively coupled plasma mass spectrometry (ICP-MS) after acid digestion in a high pressure asher (HPA-S) is presented. The digestion technique is similar to that using Carius tubes but easier to handle and reaches higher temperatures. Osmium can be determined as OsO4 with ICP-MS directly after digestion through a sparging technique. The remaining elements are preconcentrated by means of anion column chromatography. The resin is digested directly without elution leading to high yields but this causes problems if Zr is present at higher levels in the silicate rich materials. The analytical results for international platinum group element (PGE) reference materials, chromitite CHR-Bkg, basalt TDB-1 and gabbro WGB-1, are presented and compared with literature data, demonstrating the validity of the described method. Although higher in concentration, PGEs determined for reference material WGB-1 were worse than for TDB-1 indicating a more inhomogeneous distribution of the platinum group mineral phases. The low PGE abundance chromitite standard, CHR-Bkg, is likely to be homogeneous for Ru, Re, Os and Ir and is recommended as a reference material for the study of chromitites. Detection limits (3s x total procedure blank) range from 0.012 ng (Re and Os) to 0.77 ng (Pt), which could be further improved by applying higher quality acids. PMID:11284333

  16. Comparison of digestion procedures and methods for quantification of trace lead in breast milk by isotope dilution inductively coupled plasma mass spectrometry

    PubMed Central

    Amarasiriwardena, Chitra J.; Jayawardene, Innocent; Lupoli, Nicola; Barnes, Ramon M.; Hernandez-Avila, Mauricio; Hu, Howard

    2014-01-01

    Measurement of lead in breast milk is an important public health consideration and can be technically quite challenging. The reliable and accurate determination of trace lead in human breast milk is difficult for several reasons including: potential for contamination during sample collection, storage, and analysis; complexities related to the high fat content of human milk; and poor analytic sensitivity at low concentrations. Breast milk lead levels from previous published studies should therefore be reviewed with caution. Due to the difficulty in identifying a method that would successfully digest samples with 100% efficiency, we evaluated three different digestion procedures including: (1) dry ashing in a muffle furnace, (2) microwave oven digestion, and (3) digestion in high pressure asher. High temperature, high pressure asher digestion was selected as the procedure of choice for the breast milk samples. Trace lead analysis was performed using isotope dilution (ID) inductively coupled plasma mass spectrometry (ICP-MS). Measured lead concentrations in breast milk samples (n = 200) from Mexico ranged from 0.2 to 6.7 ng ml−1. The precision for these measurements ranged from 0.27–7.8% RSD. Use of strict contamination control techniques and of a very powerful digestion procedure, along with an ID-ICP-MS method for lead determination, enables us to measure trace lead levels as low as 0.2 ng ml−1 in milk (instrument detection limit = 0.01 ng ml−1). PMID:24808927

  17. Development and validation of a liquid chromatography isotope dilution mass spectrometry method for the reliable quantification of alkylphenols in environmental water samples by isotope pattern deconvolution.

    PubMed

    Fabregat-Cabello, Neus; Sancho, Juan V; Vidal, Andreu; González, Florenci V; Roig-Navarro, Antoni Francesc

    2014-02-01

    We present here a new measurement method for the rapid extraction and accurate quantification of technical nonylphenol (NP) and 4-t-octylphenol (OP) in complex matrix water samples by UHPLC-ESI-MS/MS. The extraction of both compounds is achieved in 30min by means of hollow fiber liquid phase microextraction (HF-LPME) using 1-octanol as acceptor phase, which provides an enrichment (preconcentration) factor of 800. On the other hand we have developed a quantification method based on isotope dilution mass spectrometry (IDMS) and singly (13)C1-labeled compounds. To this end the minimal labeled (13)C1-4-(3,6-dimethyl-3-heptyl)-phenol and (13)C1-t-octylphenol isomers were synthesized, which coelute with the natural compounds and allows the compensation of the matrix effect. The quantification was carried out by using isotope pattern deconvolution (IPD), which permits to obtain the concentration of both compounds without the need to build any calibration graph, reducing the total analysis time. The combination of both extraction and determination techniques have allowed to validate for the first time a HF-LPME methodology at the required levels by legislation achieving limits of quantification of 0.1ngmL(-1) and recoveries within 97-109%. Due to the low cost of HF-LPME and total time consumption, this methodology is ready for implementation in routine analytical laboratories. PMID:24423386

  18. Comparison of digestion procedures and methods for quantification of trace lead in breast milk by isotope dilution inductively coupled plasma mass spectrometry.

    PubMed

    Amarasiriwardena, Chitra J; Jayawardene, Innocent; Lupoli, Nicola; Barnes, Ramon M; Hernandez-Avila, Mauricio; Hu, Howard; Ettinger, Adrienne S

    2013-01-01

    Measurement of lead in breast milk is an important public health consideration and can be technically quite challenging. The reliable and accurate determination of trace lead in human breast milk is difficult for several reasons including: potential for contamination during sample collection, storage, and analysis; complexities related to the high fat content of human milk; and poor analytic sensitivity at low concentrations. Breast milk lead levels from previous published studies should therefore be reviewed with caution. Due to the difficulty in identifying a method that would successfully digest samples with 100% efficiency, we evaluated three different digestion procedures including: (1) dry ashing in a muffle furnace, (2) microwave oven digestion, and (3) digestion in high pressure asher. High temperature, high pressure asher digestion was selected as the procedure of choice for the breast milk samples. Trace lead analysis was performed using isotope dilution (ID) inductively coupled plasma mass spectrometry (ICP-MS). Measured lead concentrations in breast milk samples (n = 200) from Mexico ranged from 0.2 to 6.7 ng ml(-1). The precision for these measurements ranged from 0.27-7.8% RSD. Use of strict contamination control techniques and of a very powerful digestion procedure, along with an ID-ICP-MS method for lead determination, enables us to measure trace lead levels as low as 0.2 ng ml(-1) in milk (instrument detection limit = 0.01 ng ml(-1)). PMID:24808927

  19. Stability-Indicating UPLC Method for Tramadol HCl Impurities in the Tramadol Injection after Dilution by Infusion Fluids (5% Dextrose and 0.9% Sodium Chloride)

    PubMed Central

    Binnor, Anil K.; Mukkanti, Khagga; Suryanarayana, Mulukutla V.; Roy, Sunilendu B.

    2013-01-01

    A novel, rapid, and sensitive ultra-performance liquid chromatography (UPLC) method has been developed and validated as per ICH guidelines for the determination of tramadol HCl impurities in the tramadol HCl injection after reconstitution by infusion fluids (5% dextrose and 0.9% sodium chloride). The tramadol HCl injection is for the treatment of patients with moderate-to-severe pain. The stability of the reconstituted solution is critical before intravenous injection. The literature search resulted in few published articles on assays of tramadol in infusion fluids by conventional HPLC. No attempts have yet been made to determine the impurities in infusion fluids, as the concentration of tramadol after reconstitution is extremely low (0.4 mg/mL) and that of impurities is even lower. The proposed method is novel as it allows the quantitation of the impurities of tramadol HCl and is based on modern chromatographic techniques like UPLC. The method was developed using the Waters Acquity BEH C18 column with a mobile phase consisting of a gradient mixture of solvent A (trifluroacetic acid buffer) and solvent B (methanol: acetonitrile). The model stability study was designed by diluting the tramadol HCl injection in the 5% dextrose injection and 0.9% sodium chloride injection. Each mixture was kept under storage at room temperature (25 ± 2°C) for testing at initial, 2, 4, 8, 12, 18 & 24 hours. The validation study illustrates that the proposed method is suitable for the determination of tramadol and its impurities. The proposed method makes use of the LC-MS-compatible mobile phase. It can be useful for the determination of tramadol HCl and its impurities in plasma samples and other pharmaceutical dosage forms. PMID:24482769

  20. Rapid and sensitive method for the determination of four EU marker polycyclic aromatic hydrocarbons in cereal-based foods using isotope-dilution GC/MS.

    PubMed

    Kacmaz, Sibel; Zelinkova, Zuzana; Wenzl, Thomas

    2016-01-01

    A rapid and sensitive method has been developed for the determination of the four European Union marker polycyclic aromatic hydrocarbons (PAHs; benz[a]anthracene, chrysene, benzo[b]fluoranthene and benzo[a]pyrene) in some cereal-based foods. The method is based on pressurised liquid extraction (PLE), solid-phase extraction clean-up (SPE) and isotope-dilution gas chromatography with mass-spectrometric detection (GC/MS). The developed method was calibrated for the content range of 0.05-12.5 µg kg(-1) (expressed on a product basis). Recoveries of PAH were monitored in each sample via the recovery of (13)C-labelled PAHs. Recovery values were in the range between 86% and 91%, with relative standard deviations (RSDs) between 5% and 9%. The achieved limits of detection for all analytes were below 0.05 µg kg(-1). The applicability of the method for the analysis of routine samples was studied by the analysis of a set of commercial bread and breakfast cereal samples. In all analysed samples, benzo[a]pyrene (BAP) was the most prevalent PAH with the content between 0.09 and 0.30 µg kg(-1). On average, samples showed low levels of the sum of the four EU marker PAHs (ΣPAH4) that ranged between 0.11 and 0.22 µg kg(-1) for bread samples and between 0.23 and 0.87 µg kg(-1) for breakfast cereal samples. The developed method was found suitable for the determination of PAHs in cereal-based foods like cornflakes and breads with total relative fat contents below 3.5%. PMID:26950570

  1. Evaluation of agar-based medium with sheep sera for testing of drug susceptibility of Mycobacterium tuberculosis to Isoniazid, Rifampin, Ethambutol, and Streptomycin.

    PubMed

    Coban, Ahmet Yilmaz; Uzun, Meltem; Bozdogan, Bulent

    2013-12-01

    The performance of sheep sera instead of sheep blood in agar-based media was investigated for susceptibility testing of Mycobacterium tuberculosis against primary drugs. The levels of agreement between agar-based medium supplemented with sheep sera and the proportion method on Middlebrook 7H11 agar as the reference method for determining susceptibility to isoniazid (INH), rifampin (RIF), ethambutol (EMB), and streptomycin (STR) were 98.4, 98.4, 95.3, and 100%, respectively.

  2. AgarTrap-mediated genetic transformation using intact gemmae/gemmalings of the liverwort Marchantia polymorpha L.

    PubMed

    Tsuboyama-Tanaka, Shoko; Kodama, Yutaka

    2015-03-01

    The dioecious liverwort, Marchantia polymorpha L., is an emerging model plant. Various molecular biological techniques have been optimized for M. polymorpha for the past several years, and recently we reported a simplified Agrobacterium-mediated transformation method using sporelings (immature thalli from spores) of M. polymorpha. This method, termed AgarTrap (Agar-utilized Transformation with Pouring Solutions), completed by exchanging appropriate solutions on a single Petri dish to produce a sufficient number of independent transgenic sporelings. However, because spores are produced by crosses between males and females, the genetic backgrounds of resulting transgenic sporelings are not uniform. To easily produce transgenic liverworts with a uniform genetic background using AgarTrap, we developed an AgarTrap-mediated transformation method using intact gemmae/gemmalings produced by asexual reproduction. Using AgarTrap with male and female gemmae/gemmalings produced a sufficient number of independent transgenic gemmalings with uniform genetic backgrounds. The optimized transformation efficiencies were approximately 30 and 50 % in males and females, respectively. As with AgarTrap using sporelings, AgarTrap using intact gemmae/gemmalings will be useful in promoting studies of the molecular biology of M. polymorpha. PMID:25663453

  3. Development and validation of a stable-isotope dilution liquid chromatography-tandem mass spectrometry method for the determination of bisphenols in ready-made meals.

    PubMed

    Regueiro, Jorge; Wenzl, Thomas

    2015-10-01

    Due to their growing consumption, ready-made meals are a major dietary component for many people in today's society, representing an important potential route of human exposure to several food contaminants. The recent restrictions in the use of bisphenol A have led the plastic industry to look for alternative chemicals, most of them belonging to the same family of p,p'-bisphenols. The aim of the current work was to develop and validate a method based on stable-isotope dilution liquid chromatography-tandem mass spectrometry for the analysis of bisphenol A and its main analogs - bisphenol S, 4,4'-sulfonylbis(2-methylphenol), bisphenol F, bisphenol E, bisphenol B, bisphenol Z, bisphenol AF, bisphenol AP, tetrabromobisphenol A and bisphenol P - in solid foodstuffs, and particularly in ready-made meals. Extraction was carried out by ultrasound-assisted extraction after sample disruption with sand. A selective solid-phase extraction procedure was then applied to reduce potential matrix interferences. Derivatization of bisphenols with pyridine-3-sulfonyl chloride increased their ionization efficiency by electrospray ionization. Validation of the proposed method was performed in terms of selectivity, matrix effects, linearity, precision, measurement uncertainty, trueness and limits of detection. Satisfactory repeatability and intermediate precision were obtained; the related relative standard deviations were ≤7.8% and ≤10%, respectively. The relative expanded uncertainty (k=2) was below 17% for all bisphenol analogs and the trueness of the method was demonstrated by spike recovery experiments. Low limits of detection, in the range from 0.025μgkg(-1) to 0.140μgkg(-1), were obtained for all compounds. To demonstrate the applicability of the proposed method, it was eventually applied to several ready-made meals purchased from different supermarkets in Belgium. PMID:26456223

  4. Development and validation of a stable-isotope dilution liquid chromatography-tandem mass spectrometry method for the determination of bisphenols in ready-made meals.

    PubMed

    Regueiro, Jorge; Wenzl, Thomas

    2015-10-01

    Due to their growing consumption, ready-made meals are a major dietary component for many people in today's society, representing an important potential route of human exposure to several food contaminants. The recent restrictions in the use of bisphenol A have led the plastic industry to look for alternative chemicals, most of them belonging to the same family of p,p'-bisphenols. The aim of the current work was to develop and validate a method based on stable-isotope dilution liquid chromatography-tandem mass spectrometry for the analysis of bisphenol A and its main analogs - bisphenol S, 4,4'-sulfonylbis(2-methylphenol), bisphenol F, bisphenol E, bisphenol B, bisphenol Z, bisphenol AF, bisphenol AP, tetrabromobisphenol A and bisphenol P - in solid foodstuffs, and particularly in ready-made meals. Extraction was carried out by ultrasound-assisted extraction after sample disruption with sand. A selective solid-phase extraction procedure was then applied to reduce potential matrix interferences. Derivatization of bisphenols with pyridine-3-sulfonyl chloride increased their ionization efficiency by electrospray ionization. Validation of the proposed method was performed in terms of selectivity, matrix effects, linearity, precision, measurement uncertainty, trueness and limits of detection. Satisfactory repeatability and intermediate precision were obtained; the related relative standard deviations were ≤7.8% and ≤10%, respectively. The relative expanded uncertainty (k=2) was below 17% for all bisphenol analogs and the trueness of the method was demonstrated by spike recovery experiments. Low limits of detection, in the range from 0.025μgkg(-1) to 0.140μgkg(-1), were obtained for all compounds. To demonstrate the applicability of the proposed method, it was eventually applied to several ready-made meals purchased from different supermarkets in Belgium.

  5. Microstructural and Optical properties of transition metal (Cu) doped ZnO diluted magnetic semiconductor nano thin films fabricated by sol gel method

    NASA Astrophysics Data System (ADS)

    Ozturk, Ozgur; Asikuzun, Elif; Tasci, A. Tolga; Arda, Lutfi; Demirozu Senol, Sevim; Celik, Sukru; Terzioglu, Cabir

    Undoped and Cu (Copper) doped ZnO (Zn1-xCuxO) semiconductor thin films were produced by using sol-gel method. Cu was doped 1%, 2%, 3%, 4% and 5% ratio. Methanol and monoethanolamine (MEA) were used as solvent and stabilizer. In this study, the effect of Cu doping was investigated on microstructural and optical properties of ZnO DMS thin films. XRD, SEM, AFM and UV-VIS spectrometer measurements were performed for the microstructural and optical characterization. XRD, SEM and AFM results were showed that all of Cu doped ZnO based thin films have a hexagonal structure. The grain size of Cu doped ZnO thin films and morphology of surface were changed with increasing Cu doping. The optical transmittance of transition metal (Cu) doped ZnO thin films were decreased with doping. Keywords:Diluted Magnetic Semiconductor (DMS), Thin Film, Cu-doping, Bandgap Energy, ZnO. This research has been supported by the Kastamonu University Scientific Research Projects Coordination Department under the Grant No. KU-BAP-05/2015-12 and the Scientific and Technological Research Council of Turkey (TUBITAK) Project No. 114F259.

  6. Stable isotope dilution HILIC-MS/MS method for accurate quantification of glutamic acid, glutamine, pyroglutamic acid, GABA and theanine in mouse brain tissues.

    PubMed

    Inoue, Koichi; Miyazaki, Yasuto; Unno, Keiko; Min, Jun Zhe; Todoroki, Kenichiro; Toyo'oka, Toshimasa

    2016-01-01

    In this study, we developed the stable isotope dilution hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS) technique for the accurate, reasonable and simultaneous quantification of glutamic acid (Glu), glutamine (Gln), pyroglutamic acid (pGlu), γ-aminobutyric acid (GABA) and theanine in mouse brain tissues. The quantification of these analytes was accomplished using stable isotope internal standards and the HILIC separating mode to fully correct the intramolecular cyclization during the electrospray ionization. It was shown that linear calibrations were available with high coefficients of correlation (r(2)  > 0.999, range from 10 pmol/mL to 50 mol/mL). For application of the theanine intake, the determination of Glu, Gln, pGlu, GABA and theanine in the hippocampus and central cortex tissues was performed based on our developed method. In the region of the hippocampus, the concentration levels of Glu and pGlu were significantly reduced during reality-based theanine intake. Conversely, the concentration level of GABA increased. This result showed that transited theanine has an effect on the metabolic balance of Glu analogs in the hippocampus.

  7. Antimicrobial and physical-mechanical properties of agar-based films incorporated with grapefruit seed extract.

    PubMed

    Kanmani, Paulraj; Rhim, Jong-Whan

    2014-02-15

    The use of synthetic petroleum based packaging films caused serious environmental problems due to their difficulty in recycling and poor biodegradability. Therefore, present study was aimed to develop natural biopolymer-based antimicrobial packaging films as an alternative for the synthetic packaging films. As a natural antimicrobial agent, grapefruit seed extract (GSE) has been incorporated into agar to prepare antimicrobial packaging film. The films with different concentrations of GSE were prepared by a solvent casting method and the resulting composite films were examined physically and mechanically. In addition, the films were characterized by FE-SEM, XRD, FT-IR and TGA. The incorporation of GSE caused increase in color, UV barrier, moisture content, water solubility and water vapor permeability, while decrease in surface hydrophobicity, tensile strength and elastic modulus of the films. As the concentration of GSE increased from 0.6 to 13.3 μg/mL, the physical and mechanical properties of the films were affected significantly. The addition of GSE changed film microstructure of the film, but did not influence the crystallinity of agar and thermal stability of the agar-based films. The agar/GSE films exhibited distinctive antimicrobial activity against three test food pathogens, such as Listeria monocytogenes, Bacillus cereus and Escherichia coli. These results suggest that agar/GSE films have potential to be used in an active food packaging systems for maintaining food safety and extending the shelf-life of the packaged food. PMID:24507339

  8. Preparation and characterization of agar/clay nanocomposite films: the effect of clay type.

    PubMed

    Rhim, Jong-Whan; Lee, Soo-Bin; Hong, Seok-In

    2011-04-01

    Agar-based nanocomposite films with different types of nanoclays, such as Cloisite Na+, Cloisite 30B, and Cloisite 20A, were prepared using a solvent casting method, and their tensile, water vapor barrier, and antimicrobial properties were tested. Tensile strength (TS), elongation at break (E), and water vapor permeability (WVP) of control agar film were 29.7±1.7 MPa, 45.3±9.6%, and (2.22±0.19)×10(-9) g·m/m2·s·Pa, respectively. All the film properties tested, including transmittance, tensile properties, WVP, and X-ray diffraction patterns, indicated that Cloisite Na+ was the most compatible with agar matrix. TS of the nanocomposite films prepared with 5% Cloisite Na+ increased by 18%, while WVP of the nanocomposite films decreased by 24% through nanoclay compounding. Among the agar/clay nanocomposite films tested, only agar/Cloisite 30B nanocomposite film showed a bacteriostatic function against Listeria monocytogenes.

  9. STUDIES ON THE BACTERIOPHAGE OF D'HERELLE : III. SOME OF THE FACTORS DETERMINING THE NUMBER AND SIZE OF PLAQUES OF BACTERIAL LYSIS ON AGAR.

    PubMed

    Bronfenbrenner, J J; Korb, C

    1925-09-30

    The experiments reported above confirm the fact that lytic principle is distributed in active solution in a state of indivisible units. This permits its quantitative evaluation by serial dilution, as well as by plating on agar. The latter method, however, often gives readings considerably lower than those obtained by the broth dilution method of titration. By varying the concentration of agar it has been possible to show that the discrepancy is due to adsorption of the lytic agent on agar. When the concentration of the latter is increased from 0.3 per cent to 2.5 per cent the number of plaques of lysis is reduced more than 100 times. At the same time the average size of the plaques also decreases approximately to one-tenth of the original. The size, as well as the number of plaques, has been found to depend also on the condition of the culture employed in titration. Thus, when the culture exposed to the action of lytic agent is composed of young susceptible bacteria, the greater the concentration of bacteria, the smaller the plaques. When the culture is composed partly of young and partly of old susceptible bacteria, both the size and the number of the plaques are diminished with the increase in the relative concentration of old bacteria. On the other hand, presence in the culture of resistant bacteria does not affect either the size or the number of the plaques so long as the relative concentration of susceptible bacteria in the culture is sufficient to allow formation of them. The plaques appearing in the presence of a high concentration of resistant variants in the culture are relatively indistinct owing to overgrowth. Under carefully controlled conditions the size of plaques is found to be determined by the character of the lytic filtrate. Thus in the case of lytic agents which act upon more than one bacterial species the size of the plaques remains constant, irrespective of the bacterial substratum used for the production of the active filtrate.

  10. Potato Dextrose Agar Antifungal Susceptibility Testing for Yeasts and Molds: Evaluation of Phosphate Effect on Antifungal Activity of CMT-3

    PubMed Central

    Liu, Yu; Tortora, George; Ryan, Maria E.; Lee, Hsi-Ming; Golub, Lorne M.

    2002-01-01

    The broth macrodilution method (BMM) for antifungal susceptibility testing, approved by the National Committee for Clinical Laboratory Standards (NCCLS), was found to have deficiencies in testing of the antifungal activity of a new type of antifungal agent, a nonantibacterial chemically modified tetracycline (CMT-3). The high content of phosphate in the medium was found to greatly increase the MICs of CMT-3. To avoid the interference of phosphate in the test, a new method using potato dextrose agar (PDA) as a culture medium was developed. Eight strains of fungi, including five American Type Culture Collection strains and three clinical isolates, were used to determine the MICs of amphotericin B and itraconazole with both the BMM and the PDA methods. The MICs of the two antifungal agents determined with the PDA method showed 99% agreement with those determined with the BMM method within 1 log2 dilution. Similarly, the overall reproducibility of the MICs with the PDA method was above 97%. Three other antifungal agents, fluconazole, ketoconazole, and CMT-3, were also tested in parallel against yeasts and molds with both the BMM and the PDA methods. The MICs of fluconazole and ketoconazole determined with the PDA method showed 100% agreement within 1 log2 dilution of those obtained with the BMM method. However, the MICs of CMT-3 determined with the BMM method were as high as 128 times those determined with the PDA method. The effect of phosphate on the antifungal activity of CMT-3 was evaluated by adding Na2HPO4 to PDA in the new method. It was found that the MIC of CMT-3 against a Penicillium sp. increased from 0.5 μg/ml (control) to 2.0 μg/ml when the added phosphate was used at a concentration of 0.8 mg/ml, indicating a strong interference of Na2HPO4 with the antifungal activity of CMT-3. Except for fluconazole, all the other antifungal agents demonstrated clear end points among the yeasts and molds tested. Nevertheless, with its high reproducibility, good

  11. [Presumptive identification of Candida spp. and other clinically important yeasts: usefulness of Brilliance Candida Agar].

    PubMed

    Alfonso, Claudia; López, Mónica; Arechavala, Alicia; Perrone, María Del Carmen; Guelfand, Liliana; Bianchi, Mario

    2010-06-30

    Fungal infections caused by yeasts have increased during the last decades and invasive forms represent a serious problem for human health. Candida albicans is the species most frequently isolated from clinical samples. However, other emerging yeast pathogens are increasingly responsible for mycotic infections, and some of them are resistant to some antifungal drugs. Consequently, it is necessary to have methods that can provide a rapid presumptive identification at species level. Numerous chromogenic agar media have been shown to be of value as diagnostic tools. We have compared a chromogenic medium, Brilliance Candida Agar, with CHROMagar Candida, the chromogenic medium most used in our country. A multicentre study was conducted in 16 Hospitals belonging to the Mycology Net of Buenos Aires City Government. A total of 240 yeast isolates were included in this research. The new chromogenic agar showed results very similar to those obtained with CHROMagar Candida.

  12. Removal of 2,4-dinitrotoluene from concrete using bioremediation, agar extraction, and photocatalysis.

    PubMed

    Phutane, S R; Renner, J N; Nelson, S L; Seames, W S; Páca, J; Sundstrom, T J; Kozliak, E I

    2007-01-01

    Three methods, i.e. bioremediation by application of bacteria-laden agar, physical absorption of DNT by agar, or illumination by UV light were evaluated for the removal of 2,4-dinitrotoluene (DNT) from building-grade concrete. DNT biodegradation by Pseudomonas putida TOD was turned "on" and "off" by using toluene as a co-substrate thus allowing for rate-limiting step assessment. Bioremediation efficiency can be > 95-97% in 5-7 d if the process occurs at optimum growth temperature with the biological processes appearing to be rate-limiting. Sterile agar can remove up to 80% of DNT from concrete thus allowing DNT desorption and biodegradation to be conducted separately. Photoremediation results in 50% DNT removal in 9-12 d with no further removal, most likely due to mass transfer limitations.

  13. Homogeneous Matrix Deposition on Dried Agar for MALDI Imaging Mass Spectrometry of Microbial Cultures

    NASA Astrophysics Data System (ADS)

    Hoffmann, Thomas; Dorrestein, Pieter C.

    2015-11-01

    Matrix deposition on agar-based microbial colonies for MALDI imaging mass spectrometry is often complicated by the complex media on which microbes are grown. This Application Note demonstrates how consecutive short spray pulses of a matrix solution can form an evenly closed matrix layer on dried agar. Compared with sieving dry matrix onto wet agar, this method supports analyte cocrystallization, which results in significantly more signals, higher signal-to-noise ratios, and improved ionization efficiency. The even matrix layer improves spot-to-spot precision of measured m/z values when using TOF mass spectrometers. With this technique, we established reproducible imaging mass spectrometry of myxobacterial cultures on nutrient-rich cultivation media, which was not possible with the sieving technique.

  14. A universal SI-traceable isotope dilution mass spectrometry method for protein quantitation in a matrix by tandem mass tag technology.

    PubMed

    Li, Jiale; Wu, Liqing; Jin, Youxun; Su, Ping; Yang, Bin; Yang, Yi

    2016-05-01

    Isotope dilution mass spectrometry (IDMS), an important metrological method, is widely used for absolute quantification of peptides and proteins. IDMS employs an isotope-labeled peptide or protein as an internal standard although the use of a protein provides improved accuracy. Generally, the isotope-labeled protein is obtained by stable isotope labeling by amino acids in cell culture (SILAC) technology. However, SILAC is expensive, laborious, and time-consuming. To overcome these drawbacks, a novel universal SI-traceable IDMS method for absolute quantification of proteins in a matrix is described with human transferrin (hTRF). The hTRF and a human serum sample were labeled with different tandem mass tags (TMTs). After mixing the TMT-labeled hTRF and serum sample together followed by digestion, the peptides were separated by nano-liquid chromatography and analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Using the signature peptides, we calculated the ratios of reporter ions from the TMT-labeled peptides which, in turn, allowed determination of the mass fraction of hTRF. The recovery ranged from 97% to 105% with a CV of 3.9%. The LOD and LOQ were 1.71 × 10(-5) g/g and 5.69 × 10(-5) g/g of hTRF in human serum, respectively, and the relative expanded uncertainty was 4.7% with a mass fraction of 2.08 mg/g. For comparison, an enzyme-linked immunosorbent assay (ELISA) method for hTRF yielded a mass fraction of 2.03 mg/g. This method provides a starting point for establishing IDMS technology to accurately determine the mass fractions of protein biomarkers in a matrix with traceability to SI units. This technology should support the development of a metrological method useful for quantification of a wide variety of proteins. PMID:26942737

  15. Comparison of inhibitory mold agar to Sabouraud dextrose agar as a primary medium for isolation of fungi.

    PubMed

    Scognamiglio, Theresa; Zinchuk, Riva; Gumpeni, Pramod; Larone, Davise H

    2010-05-01

    Clinical specimens cultured on two selective fungal media, inhibitory mold agar (IMA) and Sabouraud dextrose agar (SDA), were compared with respect to recovery of fungi. Of the 840 fungal isolates recovered, 69.3% grew on both IMA and SDA; 24.9% grew only on IMA; and 5.8% grew only on SDA, showing that IMA is superior (P=0.003).

  16. An LC/MS/MS method for stable isotope dilution studies of β-carotene bioavailability, bioconversion, and vitamin A status in humans[S

    PubMed Central

    Oxley, Anthony; Berry, Philip; Taylor, Gordon A.; Cowell, Joseph; Hall, Michael J.; Hesketh, John; Lietz, Georg; Boddy, Alan V.

    2014-01-01

    Isotope dilution is currently the most accurate technique in humans to determine vitamin A status and bioavailability/bioconversion of provitamin A carotenoids such as β-carotene. However, limits of MS detection, coupled with extensive isolation procedures, have hindered investigations of physiologically-relevant doses of stable isotopes in large intervention trials. Here, a sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) analytical method was developed to study the plasma response from coadministered oral doses of 2 mg [13C10]β-carotene and 1 mg [13C10]retinyl acetate in human subjects over a 2 week period. A reverse phase C18 column and binary mobile phase solvent system separated β-carotene, retinol, retinyl acetate, retinyl linoleate, retinyl palmitate/retinyl oleate, and retinyl stearate within a 7 min run time. Selected reaction monitoring of analytes was performed under atmospheric pressure chemical ionization in positive mode at m/z 537→321 and m/z 269→93 for respective [12C]β-carotene and [12C] retinoids; m/z 547→330 and m/z 274→98 for [13C10]β-carotene and [13C5] cleavage products; and m/z 279→100 for metabolites of [13C10]retinyl acetate. A single one-phase solvent extraction, with no saponification or purification steps, left retinyl esters intact for determination of intestinally-derived retinol in chylomicrons versus retinol from the liver bound to retinol binding protein. Coadministration of [13C10]retinyl acetate with [13C10]β-carotene not only acts as a reference dose for inter-individual variations in absorption and chylomicron clearance rates, but also allows for simultaneous determination of an individual's vitamin A status. PMID:24158962

  17. An LC/MS/MS method for stable isotope dilution studies of β-carotene bioavailability, bioconversion, and vitamin A status in humans.

    PubMed

    Oxley, Anthony; Berry, Philip; Taylor, Gordon A; Cowell, Joseph; Hall, Michael J; Hesketh, John; Lietz, Georg; Boddy, Alan V

    2014-02-01

    Isotope dilution is currently the most accurate technique in humans to determine vitamin A status and bioavailability/bioconversion of provitamin A carotenoids such as β-carotene. However, limits of MS detection, coupled with extensive isolation procedures, have hindered investigations of physiologically-relevant doses of stable isotopes in large intervention trials. Here, a sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) analytical method was developed to study the plasma response from coadministered oral doses of 2 mg [(13)C10]β-carotene and 1 mg [(13)C10]retinyl acetate in human subjects over a 2 week period. A reverse phase C18 column and binary mobile phase solvent system separated β-carotene, retinol, retinyl acetate, retinyl linoleate, retinyl palmitate/retinyl oleate, and retinyl stearate within a 7 min run time. Selected reaction monitoring of analytes was performed under atmospheric pressure chemical ionization in positive mode at m/z 537→321 and m/z 269→93 for respective [(12)C]β-carotene and [(12)C] retinoids; m/z 547→330 and m/z 274→98 for [(13)C10]β-carotene and [(13)C5] cleavage products; and m/z 279→100 for metabolites of [(13)C10]retinyl acetate. A single one-phase solvent extraction, with no saponification or purification steps, left retinyl esters intact for determination of intestinally-derived retinol in chylomicrons versus retinol from the liver bound to retinol binding protein. Coadministration of [(13)C10]retinyl acetate with [(13)C10]β-carotene not only acts as a reference dose for inter-individual variations in absorption and chylomicron clearance rates, but also allows for simultaneous determination of an individual's vitamin A status. PMID:24158962

  18. Helium dilution refrigeration system

    DOEpatents

    Roach, Patrick R.; Gray, Kenneth E.

    1988-01-01

    A helium dilution refrigeration system operable over a limited time period, and recyclable for a next period of operation. The refrigeration system is compact with a self-contained pumping system and heaters for operation of the system. A mixing chamber contains .sup.3 He and .sup.4 He liquids which are precooled by a coupled container containing .sup.3 He liquid, enabling the phase separation of a .sup.3 He rich liquid phase from a dilute .sup.3 He-.sup.4 He liquid phase which leads to the final stage of a dilution cooling process for obtaining low temperatures. The mixing chamber and a still are coupled by a fluid line and are maintained at substantially the same level with the still cross sectional area being smaller than that of the mixing chamber. This configuration provides maximum cooling power and efficiency by the cooling period ending when the .sup.3 He liquid is depleted from the mixing chamber with the mixing chamber nearly empty of liquid helium, thus avoiding unnecessary and inefficient cooling of a large amount of the dilute .sup.3 He-.sup.4 He liquid phase.

  19. Helium dilution refrigeration system

    DOEpatents

    Roach, P.R.; Gray, K.E.

    1988-09-13

    A helium dilution refrigeration system operable over a limited time period, and recyclable for a next period of operation is disclosed. The refrigeration system is compact with a self-contained pumping system and heaters for operation of the system. A mixing chamber contains [sup 3]He and [sup 4]He liquids which are precooled by a coupled container containing [sup 3]He liquid, enabling the phase separation of a [sup 3]He rich liquid phase from a dilute [sup 3]He-[sup 4]He liquid phase which leads to the final stage of a dilution cooling process for obtaining low temperatures. The mixing chamber and a still are coupled by a fluid line and are maintained at substantially the same level with the still cross sectional area being smaller than that of the mixing chamber. This configuration provides maximum cooling power and efficiency by the cooling period ending when the [sup 3]He liquid is depleted from the mixing chamber with the mixing chamber nearly empty of liquid helium, thus avoiding unnecessary and inefficient cooling of a large amount of the dilute [sup 3]He-[sup 4]He liquid phase. 2 figs.

  20. Dilution, Concentration, and Flotation

    ERIC Educational Resources Information Center

    Liang, Ling; Schmuckler, Joseph S.

    2004-01-01

    As both classroom teaching practice and literature show, many students have difficulties learning science concepts such as density. Here are some investigations that identify the relationship between density and floating through experimenting with successive dilution of a liquid, or the systematic change of concentration of a saltwater solution.…

  1. Calibration graphs in isotope dilution mass spectrometry.

    PubMed

    Pagliano, Enea; Mester, Zoltán; Meija, Juris

    2015-10-01

    Isotope-based quantitation is routinely employed in chemical measurements. Whereas most analysts seek for methods with linear theoretical response functions, a unique feature that distinguishes isotope dilution from many other analytical methods is the inherent possibility for a nonlinear theoretical response curve. Most implementations of isotope dilution calibration today either eliminate the nonlinearity by employing internal standards with markedly different molecular weight or they employ empirical polynomial fits. Here we show that the exact curvature of any isotope dilution curve can be obtained from three-parameter rational function, y = f(q) = (a0 + a1q)/(1 + a2q), known as the Padé[1,1] approximant. The use of this function allows eliminating an unnecessary source of error in isotope dilution analysis when faced with nonlinear calibration curves. In addition, fitting with Padé model can be done using linear least squares.

  2. Evaluation of Bio-Rad MRSASelect agar for detection of methicillin-resistant Staphylococcus aureus directly from blood cultures.

    PubMed

    Riedel, Stefan; Dam, Lisa; Stamper, Paul D; Shah, Syed A R; Carroll, Karen C

    2010-06-01

    MRSASelect agar (Bio-Rad, Redmond, WA) was evaluated for its performance in detecting MRSA directly from positive blood cultures containing Gram-positive cocci in clusters. Agar plates were evaluated for the presence of pink colonies at 18 to 24 h. Results were compared to organism identification by using standard laboratory methods. Confirming coagulase on pink isolates, the sensitivity and specificity were both 99%.

  3. Flux balance analysis accounting for metabolite dilution.

    PubMed

    Benyamini, Tomer; Folger, Ori; Ruppin, Eytan; Shlomi, Tomer

    2010-01-01

    Flux balance analysis is a common method for predicting steady-state flux distributions within metabolic networks, accounting for the growth demand for the synthesis of a predefined set of essential biomass precursors. Ignoring the growth demand for the synthesis of intermediate metabolites required for balancing their dilution leads flux balance analysis to false predictions in some cases. Here, we present metabolite dilution flux balance analysis, which addresses this problem, resulting in improved metabolic phenotype predictions. PMID:20398381

  4. Flux balance analysis accounting for metabolite dilution.

    PubMed

    Benyamini, Tomer; Folger, Ori; Ruppin, Eytan; Shlomi, Tomer

    2010-01-01

    Flux balance analysis is a common method for predicting steady-state flux distributions within metabolic networks, accounting for the growth demand for the synthesis of a predefined set of essential biomass precursors. Ignoring the growth demand for the synthesis of intermediate metabolites required for balancing their dilution leads flux balance analysis to false predictions in some cases. Here, we present metabolite dilution flux balance analysis, which addresses this problem, resulting in improved metabolic phenotype predictions.

  5. Direct Protocol for Ambient Mass Spectrometry Imaging on Agar Culture.

    PubMed

    Angolini, Célio Fernando F; Vendramini, Pedro Henrique; Araújo, Francisca D S; Araújo, Welington L; Augusti, Rodinei; Eberlin, Marcos N; de Oliveira, Luciana Gonzaga

    2015-07-01

    Herein we describe a new protocol that allows direct mass spectrometry imaging (IMS) of agar cultures. A simple sample dehydration leads to a thin solid agar, which enables the direct use of spray-based ambient mass spectrometry techniques. To demonstrate its applicability, metal scavengers siderophores were imaged directly from agar culture of S. wadayamensis, and well resolved and intense images were obtained using both desorption electrospray ionization (DESI) and easy ambient sonic-spray ionization (EASI) with well-defined selective spatial distributions for the free and the metal-bound molecules, providing clues for their roles in cellular metabolism.

  6. Comparative evaluation of chromogenic agar medium and conventional culture system for isolation and presumptive identification of uropathogens

    PubMed Central

    Akter, Laila; Haque, Rezwana; Salam, Md. Abdus

    2014-01-01

    Objective: Urine is the most frequent specimen received for culture/sensitivity by clinical laboratories. The microbiological performance of HiCrome UTI agar medium was compared with Blood agar and MacConkey agar for isolation and presumptive identification of bacteria from urine culture. Methods: A total of 443 consecutively collected midstream and/or catheter-catch urine samples from patients attending the Islami Bank Medical College Hospital, Rajshahi, Bangladesh during January to December, 2012 were cultured. Urine samples showing pus cells ≥ 5/HPF were inoculated on to Blood agar (BA), MacConkey agar (MAC) and HiCrome UTI agar (CA) media simultaneously and incubated overnight aerobically at 370C. Rate of isolation and presumptive identification of bacterial species were compared for different media. Results: Culture yielded a total of 199 bacterial isolates from 189 (42.67%) positive plates including 179 (40.40%) unimicrobial and 10 (2.26%) polymicrobial (mixed growth of pair of bacteria) growths. Both HiCrome UTI agar and Blood agar media supported 100% growths while 151 (75.88%) growths were observed on MacConkey agar. The rate of presumptive identification was found significantly higher on HiCrome UTI agar (97.49%) than MAC agar (67.34%) (P<0.001) as primary urine culture medium. Of 199 isolates, E. coli was found to be the leading uropathogen isolated from 118 (59.30%) samples with its presumptive identification rate of 95.76%, 93.22% and 5.93% on CA, MAC and BA respectively. All 10 (100%) polymicrobial growths were demonstrated distinctly on CA against only 01(10%) on each BA and MAC. Conclusion: HiCrome UTI agar was found to be more useful as primary urine culture medium in both higher rate of isolation and presumptive identification of uropathogens in comparison to conventional media. Its inherent characteristics in demonstrating polymicrobial growth and ease of rapid identification by distinct colony colour are unique. PMID:25225521

  7. Evaluation of different methods for the detection of methicillin resistance in coagulase-negative staphylococci.

    PubMed

    Jarløv, J O; Busch-Sørensen, C; Espersen, F; Mortensen, I; Hougaard, D M; Rosdahl, V T

    1997-08-01

    The efficacy of 19 agar diffusion methods for the detection of methicillin resistance among coagulase-negative staphylococci (CoNS) within 24 h was evaluated. A total of 359 CoNS isolates were tested, of which 204 were Staphylococcus epidermidis. In 164 isolates, the presence of mecA was investigated; 61 strains were mecA-positive and 103 were mecA-negative by Southern blot analysis. Based on the best agreement shown with the mecA determination (94%) among four agar dilution assays for determining methicillin MIC, an assay with Columbia agar supplemented with NaCl and incubation with a heavy bacterial inoculum of 10(5)-10(6) cfu/spot was used as the reference MIC method. The best agar diffusion results were obtained with a 1 microg oxacillin disc on Columbia agar with 4.5% NaCl supplement. With this method, 99% of S. epidermidis and 94% of non-S. epidermidis were in agreement with the MIC determination. However, Columbia (without NaCl), Mueller-Hinton and Isosensitest agars were almost as useful when a 1 microg oxacillin disc was used. The zone breakpoints for S. epidermidis were, in general, considerably larger than those for other CoNS species and, consequently, differentiation according to species is recommended. Furthermore, resistance to other antibiotics, such as gentamicin and erythromycin, makes methicillin resistance highly likely.

  8. Evaluation of different methods for the detection of methicillin resistance in coagulase-negative staphylococci.

    PubMed

    Jarløv, J O; Busch-Sørensen, C; Espersen, F; Mortensen, I; Hougaard, D M; Rosdahl, V T

    1997-08-01

    The efficacy of 19 agar diffusion methods for the detection of methicillin resistance among coagulase-negative staphylococci (CoNS) within 24 h was evaluated. A total of 359 CoNS isolates were tested, of which 204 were Staphylococcus epidermidis. In 164 isolates, the presence of mecA was investigated; 61 strains were mecA-positive and 103 were mecA-negative by Southern blot analysis. Based on the best agreement shown with the mecA determination (94%) among four agar dilution assays for determining methicillin MIC, an assay with Columbia agar supplemented with NaCl and incubation with a heavy bacterial inoculum of 10(5)-10(6) cfu/spot was used as the reference MIC method. The best agar diffusion results were obtained with a 1 microg oxacillin disc on Columbia agar with 4.5% NaCl supplement. With this method, 99% of S. epidermidis and 94% of non-S. epidermidis were in agreement with the MIC determination. However, Columbia (without NaCl), Mueller-Hinton and Isosensitest agars were almost as useful when a 1 microg oxacillin disc was used. The zone breakpoints for S. epidermidis were, in general, considerably larger than those for other CoNS species and, consequently, differentiation according to species is recommended. Furthermore, resistance to other antibiotics, such as gentamicin and erythromycin, makes methicillin resistance highly likely. PMID:9301990

  9. Evaluation and interlaboratory validation of a selective agar for phosphatidylinositol-specific phospholipase C activity using a chromogenic substrate to detect Listeria monocytogenes from foods.

    PubMed

    Jinneman, Karen C; Hunt, Jan M; Eklund, Cheryl A; Wernberg, Jane S; Sado, Patricia N; Johnson, Janelle M; Richter, Richelle S; Torres, Selene T; Ayotte, Eugene; Eliasberg, Stacey J; Istafanos, Phillip; Bass, Deborah; Kexel-Calabresa, Nancy; Lin, Wen; Barton, Curtis N

    2003-03-01

    Phosphatidylinositol-specific phospholipase C (PI-PLC) activity is a potential virulence factor and is exhibited only by the Listeria species Listeria monocytogenes and Listeria ivanovii. A chromogenic substrate for the direct detection of PI-PLC activity is available in a new medium (BCM L. monocytogenes plating agar). The use of a chromogenic substrate offers a mechanism with which to directly screen for L. monocytogenes and L. ivanovii other than the esculin used in Oxford (OXF) and Palcam (PAL) agars, which screen for all Listeria species. The specificity levels of BCM plating agar and of BCM confirmation and rhamnose agars were evaluated with 107 Listeria and 10 Bacillus species isolates. In addition, BCM L. monocytogenes plating agar was compared with standard Listeria selective agars (OXF and PAL agars) with regard to the recovery of L. monocytogenes from 2,000 food and environmental samples obtained from eight participating laboratories. A Listeria species was isolated from at least one of the agars in 209 analyses, and L. monocytogenes was isolated in 135 of these analyses. In 27 of the analyses in which L. monocytogenes was isolated, one or more of the selective differential agars used failed to isolate L. monocytogenes, and therefore the results of these analyses were discrepant. Relative to a reference method involving the use of all three agars (OXF, PAL, and BCM agars), the OXF-BCM, PAL-BCM, and OXF-PAL combinations had sensitivities of 99.3, 99.2, and 90.2%, respectively. In statistical analyses of the different combinations of agars, the OXF-BCM and BCM-PAL combinations were found to be superior to the OXF-PAL combination for the detection of L. monocytogenes.

  10. Preparation of an agar-silver nanoparticles (A-AgNp) film for increasing the shelf-life of fruits.

    PubMed

    Gudadhe, Janhavi A; Yadav, Alka; Gade, Aniket; Marcato, Priscyla D; Durán, Nelson; Rai, Mahendra

    2014-12-01

    Preparation of protective coating possessing antimicrobial properties is present day need as they increase the shelf life of fruits and vegetables. In the present study, preparation of agar-silver nanoparticle film for increasing the shelf life of fruits is reported. Silver nanoparticles (Ag-NPs) biosynthesised using an extract of Ocimum sanctum leaves, were mixed with agar-agar to prepare an agar-silver nanoparticles (A-AgNp) film. This film was surface-coated over the fruits, Citrus aurantifolium (Thornless lime) and Pyrus malus (Apple), and evaluated for the determination of antimicrobial activity of A-AgNp films using disc diffusion method, weight loss and shelf life of fruits. This study demonstrates that these A-AgNp films possess antimicrobial activity and also increase the shelf life of fruits. PMID:25429496

  11. Automated counting of bacterial colony forming units on agar plates.

    PubMed

    Brugger, Silvio D; Baumberger, Christian; Jost, Marcel; Jenni, Werner; Brugger, Urs; Mühlemann, Kathrin

    2012-01-01

    Manual counting of bacterial colony forming units (CFUs) on agar plates is laborious and error-prone. We therefore implemented a colony counting system with a novel segmentation algorithm to discriminate bacterial colonies from blood and other agar plates.A colony counter hardware was designed and a novel segmentation algorithm was written in MATLAB. In brief, pre-processing with Top-Hat-filtering to obtain a uniform background was followed by the segmentation step, during which the colony images were extracted from the blood agar and individual colonies were separated. A Bayes classifier was then applied to count the final number of bacterial colonies as some of the colonies could still be concatenated to form larger groups. To assess accuracy and performance of the colony counter, we tested automated colony counting of different agar plates with known CFU numbers of S. pneumoniae, P. aeruginosa and M. catarrhalis and showed excellent performance.

  12. [Evaluation of blood agar medium for the growth of mycobacteria].

    PubMed

    Coban, Ahmet Yılmaz; Akgüneş, Alper; Durupınar, Belma

    2011-10-01

    This study was aimed to evaluate the performance of blood agar for the growth of mycobacteria from clinical specimens sent to Mycobacteriology Laboratory of Samsun Chest Diseases Hospital. One hundred fifty six clinical specimens including 123 sputum, 28 bronchoalveolar lavage (BAL) and 5 pleural fluid specimens were inoculated in Löwenstein-Jensen (LJ), BACTEC MGIT 960 system (Becton Dickinson, USA) and blood agar following decontamination process. The specimens were also simultaneously examined for the presence of acid-fast bacilli (AFB). Thirty five mycobacteria strains (33 Mycobacterium tuberculosis and 2 atypical mycobacteria) grew in blood agar, 38 (36 M.tuberculosis and 2 atypical mycobacteria) in LJ media and 46 (44 M.tuberculosis and 2 atypical mycobacteria) in BACTEC MGIT 960 system. Among 29 AFB negative specimens, 20 revealed growth in both blood agar and LJ medium and 27 in MGIT system. AFB positive 20 samples yielded growth in 15 samples in blood agar, 18 in LJ medium and 19 in MGIT system. Among the total of 156 samples, contamination was observed in 15 (9.6%) samples in blood agar, 16 (10.2%) in LJ medium and 18 (11.5%) in MGIT system. Growth time was 5-35 days (mean 18 ± 7.4), 11-35 days (mean 19 ± 5.9) and 5-15 days (mean 10 ± 2.4) for blood agar, LJ medium and BACTEC MGIT 960 system, respectively. The three samples which revealed contamination in BACTEC MGIT 960 system, grew successfully in both blood agar and LJ medium without contamination. In one sample, growth was observed only in LJ medium but neither in blood agar nor BACTEC MGIT 960 system. However, in another sample, growth was observed only in blood agar while no growth was detected in LJ or BACTEC MGIT 960 system. Six samples yielded mycobacteria only in BACTEC MGIT 960 system. These results indicated that simultaneous use of one liquid and one solid medium to grow mycobacteria from the clinical samples seemed to be complementary. Blood agar was a promising choice since it was found

  13. Use of Mueller-Hinton broth and agar in the germ tube test.

    PubMed

    Mattei, Antonella Souza; Alves, Sydney Hartz; Severo, Cecília Bittencourt; Guazzelli, Luciana da Silva; Oliveira, Flávio de Mattos; Severo, Luiz Carlos

    2014-01-01

    Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.

  14. Hyperspectral image reconstruction using RGB color for foodborne pathogen detection on agar plates

    NASA Astrophysics Data System (ADS)

    Yoon, Seung-Chul; Shin, Tae-Sung; Park, Bosoon; Lawrence, Kurt C.; Heitschmidt, Gerald W.

    2014-03-01

    This paper reports the latest development of a color vision technique for detecting colonies of foodborne pathogens grown on agar plates with a hyperspectral image classification model that was developed using full hyperspectral data. The hyperspectral classification model depended on reflectance spectra measured in the visible and near-infrared spectral range from 400 and 1,000 nm (473 narrow spectral bands). Multivariate regression methods were used to estimate and predict hyperspectral data from RGB color values. The six representative non-O157 Shiga-toxin producing Eschetichia coli (STEC) serogroups (O26, O45, O103, O111, O121, and O145) were grown on Rainbow agar plates. A line-scan pushbroom hyperspectral image sensor was used to scan 36 agar plates grown with pure STEC colonies at each plate. The 36 hyperspectral images of the agar plates were divided in half to create training and test sets. The mean Rsquared value for hyperspectral image estimation was about 0.98 in the spectral range between 400 and 700 nm for linear, quadratic and cubic polynomial regression models and the detection accuracy of the hyperspectral image classification model with the principal component analysis and k-nearest neighbors for the test set was up to 92% (99% with the original hyperspectral images). Thus, the results of the study suggested that color-based detection may be viable as a multispectral imaging solution without much loss of prediction accuracy compared to hyperspectral imaging.

  15. Use of Dehydrated Agar to Estimate Microbial Water Quality for Horticulture Irrigation.

    PubMed

    Meador, Dustin P; Fisher, Paul R; Guy, Charles L; Harmon, Philip F; Peres, Natalia A; Teplitski, Max

    2016-07-01

    Petrifilms are dehydrated agar culture plates that have been used to quantify colony forming units (CFU) mL of either aerobic bacteria (Petrifilm-AC) or fungus (Petrifilm-YM), depending on substrate composition. Microbes in irrigation systems can indicate biofilm risk and potential clogging of irrigation emitters. The research objective was to compare counts on Petrifilms versus traditional, hydrated-agar plates using samples collected from recirculated irrigation waters and cultures of isolated known species. The estimated count (in CFU mL) from a recirculated irrigation sample after 7 d of incubation on Petrifilm-YM was only 5.5% of the count quantified using sabouraud dextrose agar (SDA) with chloramphenicol after 14 d. In a separate experiment with a known species, Petrifilm-YM did not successfully culture zoospores of . Isolates of viable zoospores were cultured successfully on potato-dextrose agar (PDA), with comparable counts with a vegetable juice medium supplemented with the antibiotics pimaricin, ampicillin, rifamycin, pentochloronitrobenzene and hymexazol (PARP-H). The quantification of pv. Begoniaceae on Petrifilm-AC was not significantly different ( < 0.05) than on PDA, but was lower than on Reasoner and Goldrich agar (R2A) or with a hemocytometer. The current formulation of Petrifilm-YM is unlikely to be a useful monitoring method for plant pathogens in irrigation water because of the inability to successfully culture oomycetes. However, Petrifilm-AC was an effective method to quantify bacteria and can provide an easy-to-use on-farm tool to monitor biofilm risk and microbial density.

  16. Use of Dehydrated Agar to Estimate Microbial Water Quality for Horticulture Irrigation.

    PubMed

    Meador, Dustin P; Fisher, Paul R; Guy, Charles L; Harmon, Philip F; Peres, Natalia A; Teplitski, Max

    2016-07-01

    Petrifilms are dehydrated agar culture plates that have been used to quantify colony forming units (CFU) mL of either aerobic bacteria (Petrifilm-AC) or fungus (Petrifilm-YM), depending on substrate composition. Microbes in irrigation systems can indicate biofilm risk and potential clogging of irrigation emitters. The research objective was to compare counts on Petrifilms versus traditional, hydrated-agar plates using samples collected from recirculated irrigation waters and cultures of isolated known species. The estimated count (in CFU mL) from a recirculated irrigation sample after 7 d of incubation on Petrifilm-YM was only 5.5% of the count quantified using sabouraud dextrose agar (SDA) with chloramphenicol after 14 d. In a separate experiment with a known species, Petrifilm-YM did not successfully culture zoospores of . Isolates of viable zoospores were cultured successfully on potato-dextrose agar (PDA), with comparable counts with a vegetable juice medium supplemented with the antibiotics pimaricin, ampicillin, rifamycin, pentochloronitrobenzene and hymexazol (PARP-H). The quantification of pv. Begoniaceae on Petrifilm-AC was not significantly different ( < 0.05) than on PDA, but was lower than on Reasoner and Goldrich agar (R2A) or with a hemocytometer. The current formulation of Petrifilm-YM is unlikely to be a useful monitoring method for plant pathogens in irrigation water because of the inability to successfully culture oomycetes. However, Petrifilm-AC was an effective method to quantify bacteria and can provide an easy-to-use on-farm tool to monitor biofilm risk and microbial density. PMID:27380096

  17. Minimal Coital Dilution in Accra, Ghana

    PubMed Central

    Jenness, Samuel M.; Biney, Adriana A. E.; Ampofo, William Kwabena; Dodoo, F. Nii-Amoo; Cassels, Susan

    2015-01-01

    Background Coital dilution, the reduction in the coital frequency per partner when an additional ongoing partner is added, may reduce the transmission potential of partnership concurrency for HIV and other sexually transmitted infections. Empirical estimates of dilution, especially dilution of sexual acts unprotected by condoms, are needed to inform prevention research. Methods Sexually active adults in Accra, Ghana were recruited in a multi-stage household probability sample. Degree (number of ongoing partners), total acts, and unprotected acts were measured retrospectively for each month in the past year through an event-history calendar. Random effects negative binomial models estimated the association between degree and coital frequency. Results Compared to person-months with a single partner (monogamy), 2.06 times as many total acts and 1.94 times as many unprotected acts occurred in months with 2 partners. In months with 3 partners, 2.90 times as many total acts and 2.39 times as many unprotected acts occurred compared to monogamous months. Total acts but not unprotected acts also declined with partnership duration. Conclusions No dilution was observed for total acts with up to three concurrent partners, but a small amount of dilution was observed for unprotected acts for months with multiple concurrencies. This suggests moderate selective condom use in months with multiple concurrencies. The implications of the observed dilution for future HIV transmission must be investigated with mathematical models. PMID:25622062

  18. Gluconeogenesis from labeled carbon: estimating isotope dilution

    SciTech Connect

    Kelleher, J.K.

    1986-03-01

    To estimate the rate of gluconeogenesis from steady-state incorporation of labeled 3-carbon precursors into glucose, isotope dilution must be considered so that the rate of labeling of glucose can be quantitatively converted to the rate of gluconeogenesis. An expression for the value of this isotope dilution can be derived using mathematical techniques and a model of the tricarboxylic acid (TCA) cycle. The present investigation employs a more complex model than that used in previous studies. This model includes the following pathways that may affect the correction for isotope dilution: 1) flux of 3-carbon precursor to the oxaloacetate pool via acetyl-CoA and the TCA cycle; 2) flux of 4- or 5-carbon compounds into the TCA cycle; 3) reversible flux between oxaloacetate (OAA) and pyruvate and between OAA and fumarate; 4) incomplete equilibrium between OAA pools; and 5) isotope dilution of 3-carbon tracers between the experimentally measured pool and the precursor for the TCA-cycle OAA pool. Experimental tests are outlined which investigators can use to determine whether these pathways are significant in a specific steady-state system. The study indicated that flux through these five pathways can significantly affect the correction for isotope dilution. To correct for the effects of these pathways an alternative method for calculating isotope dilution is proposed using citrate to relate the specific activities of acetyl-CoA and OAA.

  19. Cost-effective nanoporous Agar-Agar polymer/Nickel powder composite particle for effective bio-products adsorption by expanded bed chromatography.

    PubMed

    Asgari, Setareh; Jahanshahi, Mohsen; Rahimpour, Ahmad

    2014-09-26

    In the present work a novel kind of dense nanoporous composite matrix for expanded bed application has been successfully first prepared with Nickel powder as a densifier and was covered with Agar-Agar layer as a skeleton, through the method of water-in-oil emulsification. Agar-Agar is a porous and inexpensive polymer. In order to fabricate cost-effective adsorbent with favorable qualities Agar-Agar polymer was used. Thereafter, the customized composite particle was modified by pseudo-affinity dye-ligand, Reactive Blue 4 (RB4), aimed at preparing a pseudo-affinity adsorbent (RB4-Agar-Ni) for bioprodut adsorption from aqueous solution. Bovine Serum Albumin (BSA) was selected as a model protein to investigate the adsorption behavior in batchwise and expanded bed chromatography, and the obtained results were evaluated with that of Streamline™ (Amersham-Pharmacia Biotech, Sweden). Spherical appearance and porous structure of composite particles were observed by the optical microscope (OM) and scanning electronic microscope (SEM). The results suggested that the matrices followed the logarithmic normal size distribution with the range of 65-300 μm and average diameter of 126.81-151.47 μm, proper wet density of 1.64-2.78 g/ml, water content of 62.74-34%, porosity of 98-90% and pore size of about 38-130 nm. For better comprehension of the impact of solid phase properties on the performance of the expanded bed, the expansion and hydrodynamic properties of a composite matrix with a series of densities was evaluated and estimated by the retention time distribution method (RTD) in an expanded bed and was compared with that of other matrices. According to obtained results the expansion factors under the same fluid velocity decreased by increasing the matrix density. Moreover, the axial dispersion coefficient (Dax) is the most appropriate parameter for evaluating the stability of expanded bed, on various operating conditions, such as different flow velocity, bed expansion

  20. Cost-effective nanoporous Agar-Agar polymer/Nickel powder composite particle for effective bio-products adsorption by expanded bed chromatography.

    PubMed

    Asgari, Setareh; Jahanshahi, Mohsen; Rahimpour, Ahmad

    2014-09-26

    In the present work a novel kind of dense nanoporous composite matrix for expanded bed application has been successfully first prepared with Nickel powder as a densifier and was covered with Agar-Agar layer as a skeleton, through the method of water-in-oil emulsification. Agar-Agar is a porous and inexpensive polymer. In order to fabricate cost-effective adsorbent with favorable qualities Agar-Agar polymer was used. Thereafter, the customized composite particle was modified by pseudo-affinity dye-ligand, Reactive Blue 4 (RB4), aimed at preparing a pseudo-affinity adsorbent (RB4-Agar-Ni) for bioprodut adsorption from aqueous solution. Bovine Serum Albumin (BSA) was selected as a model protein to investigate the adsorption behavior in batchwise and expanded bed chromatography, and the obtained results were evaluated with that of Streamline™ (Amersham-Pharmacia Biotech, Sweden). Spherical appearance and porous structure of composite particles were observed by the optical microscope (OM) and scanning electronic microscope (SEM). The results suggested that the matrices followed the logarithmic normal size distribution with the range of 65-300 μm and average diameter of 126.81-151.47 μm, proper wet density of 1.64-2.78 g/ml, water content of 62.74-34%, porosity of 98-90% and pore size of about 38-130 nm. For better comprehension of the impact of solid phase properties on the performance of the expanded bed, the expansion and hydrodynamic properties of a composite matrix with a series of densities was evaluated and estimated by the retention time distribution method (RTD) in an expanded bed and was compared with that of other matrices. According to obtained results the expansion factors under the same fluid velocity decreased by increasing the matrix density. Moreover, the axial dispersion coefficient (Dax) is the most appropriate parameter for evaluating the stability of expanded bed, on various operating conditions, such as different flow velocity, bed expansion

  1. Agar/collagen membrane as skin dressing for wounds.

    PubMed

    Bao, Lei; Yang, Wei; Mao, Xuan; Mou, Shansong; Tang, Shunqing

    2008-12-01

    Agar, a highly hydrophilic polymer, has a special gel property and favorable biocompatibility, but moderate intension strength in an aqueous condition and a low degradation rate. In order to tailor both properties of mechanical intension and degradation, type I collagen was composited with agar in a certain ratio by drying at 50 degrees C or by a freeze-dry process. Glutaraldehyde was chosen as a crosslinking agent, and the most favorable condition for crosslinking was that the weight ratio of agar to glutaraldehyde was 66.7 and the pH value about 5. Dynamic mechanical analysis results showed that the single agar membrane had a modulus value between 640 MPa and 1064 MPa, but it was between 340 MPa and 819 MPa after being composited with type I collagen. It was discovered under an optical microscope that the pores were interconnected in the composite scaffolds instead of the honeycomb-like pores in a single type I collagen scaffold or the laminated gaps in a single agar scaffold. The results of an acute toxicity test disclosed that the composites were not toxic to mice although the composites were crosslinked with a certain concentration of glutaraldehyde. The results of gross examinations showed that when the composite membranes or scaffolds were applied to a repair rabbit skin lesion, the composites had a good repair effect without infection, liquid exudation or visible scar in the lesion covered with them. But in the control group, the autologous skin showed necrosis and there were a lot of scar tissues in the lesion site. H&E staining results showed that the repair tissue was similar to the normal one and very few scaffolds or membranes were left without degradation after 2 or 3 weeks. In conclusion, it is proved that type I collagen increases the toughness of the agar membrane, and the agar/type I collagen composites are promising biomaterials as wound dressings for healing burns or ulcers.

  2. Production of microbial medium from defatted brebra (Milletia ferruginea) seed flour to substitute commercial peptone agar

    PubMed Central

    Andualem, Berhanu; Gessesse, Amare

    2013-01-01

    Objective To investigate and optimize microbial media that substitute peptone agar using brebra seed defatted flour. Methods 'Defatted process, inoculums preparation, evaluation of bacterial growth, preparation of cooked and hydrolyzed media and growth turbidity of tested bacteria were determined. Results Two percent defatted flour was found to be suitable concentration for the growth of pathogenic bacteria: Escherichia coli (ATCC 25922) (E. coli), Pseudomonas aeruginosa (ATCC 27853), Salmonella (NCTC 8385) and Shigella flexneri (ATCC 12022) (S. flexneri), while 3% defatted flour was suitable for Staphylococcus aureus (ATCC 25923) (S. aureus). E. coli (93±1) and S. flexneri (524±1) colony count were significantly (P≤0.05) greater in defatted flour without supplement than in supplemented medium. E. coli [(3.72×109±2) CFU/mL], S. aureus [(7.4×109±2) CFU/mL], S. flexneri [(4.03×109±2) CFU/mL] and Salmonella [(2.37×109±1) CFU/mL] in non-hydrolyzed sample were statistically (P≤0.05) greater than hydrolyzed one and commercial peptone agar. Colony count of Salmonella [(4.55×109±3) CFU/mL], S. flexneri [(5.40×109±3) CFU/mL] and Lyesria moncytogenes (ATCC 19116) [(5.4×109±3) CFU/mL] on raw defatted flour agar was significantly (P≤0.05) greater than cooked defatted flour and commercial peptone agar. Biomass of E. coli, S. aureus, Salmonella and Enterococcus faecalis in non-hydrolyzed defatted flour is highly increased over hydrolyzed defatted flour and commercial peptone broth. Conclusions The defatted flour agar was found to be better microbial media or comparable with peptone agar. The substances in it can serve as sources of carbon, nitrogen, vitamins and minerals that are essential to support the growth of microorganisms without any supplements. Currently, all supplements of peptone agar are very expensive in the market. PMID:24075344

  3. Evaluation of chromogenic agar, [corrected] VITEK2 YST and VITEK® MS for identification of Candida strains isolated from blood cultures.

    PubMed

    Sariguzel, Fatma Mutlu; Berk, Elife; Koc, Ayse Nedret; Sav, Hafize; Aydemir, Gonca

    2015-12-01

    The aim of this study is to compare conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system for the identification of Candida strains isolated from blood cultures. Fifty-four strains were identified according to conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS. Sequencing was used as the reference method. The 54 strains included 32 Candida parapsilosis, 19 Candida albicans, 1 Candida glabrata and 2 Candida tropicalis according to the reference method. One C. albicans and one C. glabrata isolate were misidentified as C. parapsilosis by chromogenic agar. [corrected]. Two C. parapsilosis and three C. albicans isolates were misidentified by VITEK2 YST card. Chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS identified correctly 96.2%, 90.7% and 100% of all strains, respectively. We found that the chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system are easy, rapid and accurate alternative methods for the identification of yeast species in the clinical microbiology laboratory.

  4. Metronidazole and clarithromycin resistance in Helicobacter pylori determined by measuring MICs of antimicrobial agents in color indicator egg yolk agar in a miniwell format. The Gastrointestinal Physiology Working Group of Universidad Peruana Cayetano Heredia and the Johns Hopkins University.

    PubMed

    Vasquez, A; Valdez, Y; Gilman, R H; McDonald, J J; Westblom, T U; Berg, D; Mayta, H; Gutierrez, V

    1996-05-01

    Resistance of Helicobacter pylori to metronidazole often causes failure of commonly used combination drug treatment regimens. We determined the MICs of metronidazole and clarithromycin against 18 H. pylori strains from Peru using tetrazolium egg yolk (TEY) agar. The MIC results obtained by agar dilution with petri dishes were compared with the results found through a miniwell format. The results of the two protocols for measuring drug susceptibility differed by no more than 1 dilution in all cases. On TEY agar, bright-red H. pylori colonies were easy to identify against a yellow background. Sixty-one percent (11 of 18) of the strains were resistant to metronidazole (MIC, > or = 4 micrograms/ml) and 50% (9 of 18) were resistant to clarithromycin (MIC, > or = 0.125 micrograms/ml), whereas none (0 of 5) of the strains tested were resistant to tetracycline (MIC, > or = 1 micrograms/ml). Thus, the prevalence of metronidazole and clarithromycin resistance in Peru is higher than that in developed regions of the world. The miniwell plate with TEY agar allows easy H. pylori colony identification, requires about one-third less of the costly medium necessary for petri dish assaying, conserves space, and yields MICs equivalent to those with agar dilution in petri dishes.

  5. Statistical inference for serial dilution assay data.

    PubMed

    Lee, M L; Whitmore, G A

    1999-12-01

    Serial dilution assays are widely employed for estimating substance concentrations and minimum inhibitory concentrations. The Poisson-Bernoulli model for such assays is appropriate for count data but not for continuous measurements that are encountered in applications involving substance concentrations. This paper presents practical inference methods based on a log-normal model and illustrates these methods using a case application involving bacterial toxins.

  6. Blood agar and Mycobacterium tuberculosis: the end of a dogma.

    PubMed

    Drancourt, M; Carrieri, P; Gévaudan, M-J; Raoult, D

    2003-04-01

    Incidental blood agar-based recovery of Mycobacterium tuberculosis led us to further investigate this routine medium for primary isolation and culture of M. tuberculosis. Fifteen respiratory tract and eight lymph node Ziehl-Neelsen-positive specimens were inoculated in parallel into tubes containing egg-based medium and 5% sheep blood agar. Colonies appeared sooner on this medium than on the egg-based medium, but this difference was not significant (P = 0.11, analysis of variance [ANOVA] test). Further experiments compared the growth of 38 respiratory and lymph node M. tuberculosis isolates when subcultured on the two media. After 6 days of incubation, 21 of 38 isolates had grown on blood agar, and the mean number of colonies was significantly greater on blood agar than on the egg-based medium (P < 0 0.001, ANOVA test). These results demonstrate that M. tuberculosis grows easily on blood agar within 1to 2 weeks, indicating that this basic medium is suitable for laboratory diagnosis of tuberculosis in addition to other media. Laboratories that routinely use prolonged incubations of blood plates, for example, for the recovery of Bartonella species, should consider the potential safety implications of encountering this highly infectious pathogen.

  7. [Stich-plaque test--an economic method for quantitative determination of viruses].

    PubMed

    Waschke, K; Stäber, H

    1976-01-01

    An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID 50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity , accuracy, and reproductibility of this method are described and discussed.

  8. [Stick-plaque test--an economic method of quantitative determination of viruses].

    PubMed

    Waschke, K; Stäber, H

    1976-01-01

    An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity, accuracy, and reproductibility of this method are described and discussed.

  9. Photothermal characterization of the gelation process in Gelidium robustum Agar

    NASA Astrophysics Data System (ADS)

    Freile-Pelegrín, Y.; Bante, J.; Alvarado-Gil, J. J.; Yánez-Limón, J. M.

    2005-06-01

    Agar is a hydrophilic colloid formed by polysaccharides, whose ability to form reversible gels simply by cooling hot aqueous solutions is the most important property and can be regarded as the prototype and model for all gelling systems. In this paper the evolution of the gelation process of agar obtained from algae of the species Gelidium robustum, using the photopyroelectric technique is reported. It is shown that thermal effusivity increase when the agar is cooled, reaching a maximum value around 37°C. The increase in thermal effusivity can be related to the increasing of the bondings in the gel as temperature decreases, reaching the maximum at the gelation point. The decrease of the thermal effusivity at lower temperature could be due to the syneresis process involving a gradual release of water after gelation.

  10. Fly agaric (Amanita muscaria) poisoning, case report and review.

    PubMed

    Satora, Leszek; Pach, Dorota; Butryn, Beata; Hydzik, Piotr; Balicka-Slusarczyk, Barbara

    2005-06-01

    Gathering and eating mushrooms and other plants containing psychoactive substances has become increasingly popular among young people experimenting with drugs. Dried fly agaric Amanita muscaria fruiting bodies were eaten by five young persons (18-21 years of age) at a party in order to evoke hallucinations. Visual and auditory hallucinations occurred in four of them, whereas a 18-year-old girl lost consciousness. The following morning, she went to the Clinic of Toxicology. Due to the fact that not all the active substances present in the fly agaric have been identified, and some of them have an effect after a period of latency, the patient was admitted for several days of observation during which check-up examinations were performed. After four days without any problems, she was discharged. The poisoning regressed with no organ complications. The remaining persons who had eaten the fly agaric were free from any complaints. PMID:15904689

  11. Agar-Block Microcosms for Controlled Plant Tissue Decomposition by Aerobic Fungi

    PubMed Central

    Schilling, Jonathan S.

    2011-01-01

    The two principal methods for studying fungal biodegradation of lignocellulosic plant tissues were developed for wood preservative testing (soil-block; agar-block). It is well-accepted that soil-block microcosms yield higher decay rates, fewer moisture issues, lower variability among studies, and higher thresholds of preservative toxicity. Soil-block testing is thus the more utilized technique and has been standardized by American Society for Testing and Materials (ASTM) (method D 1413-07). The soil-block design has drawbacks, however, using locally-variable soil sources and in limiting the control of nutrients external (exogenous) to the decaying tissues. These drawbacks have emerged as a problem in applying this method to other, increasingly popular research aims. These modern aims include degrading lignocellulosics for bioenergy research, testing bioremediation of co-metabolized toxics, evaluating oxidative mechanisms, and tracking translocated elements along hyphal networks. Soil-blocks do not lend enough control in these applications. A refined agar-block approach is necessary. Here, we use the brown rot wood-degrading fungus Serpula lacrymans to degrade wood in agar-block microcosms, using deep Petri dishes with low-calcium agar. We test the role of exogenous gypsum on decay in a time-series, to demonstrate the utility and expected variability. Blocks from a single board rip (longitudinal cut) are conditioned, weighed, autoclaved, and introduced aseptically atop plastic mesh. Fungal inoculations are at each block face, with exogenous gypsum added at interfaces. Harvests are aseptic until the final destructive harvest. These microcosms are designed to avoid block contact with agar or Petri dish walls. Condensation is minimized during plate pours and during incubation. Finally, inoculum/gypsum/wood spacing is minimized but without allowing contact. These less technical aspects of agar-block design are also the most common causes of failure and the key source of

  12. Growth of Desulfovibrio on the Surface of Agar Media

    PubMed Central

    Iverson, Warren P.

    1966-01-01

    Growth of Desulfovibrio desulfuricans (API strain) was found to take place in an atmosphere of hydrogen on the agar surface of complex media, including yeast extract (Difco), and Trypticase Soy Agar (BBL) without any added reducing agents. For growth on a 2% yeast extract-agar surface in the absence of hydrogen (nitrogen atmosphere), sodium lactate was required in the medium. Growth on the surface of Trypticase Soy Agar (TSA) under nitrogen took place readily in the absence of an added hydrogen donor. A medium (TSA plus salts) is described based upon the addition of sodium lactate (4 ml per liter), magnesium sulfate (2 g per liter), and ferrous ammonium sulfate (0.05%) to TSA, which appears suitable for the isolation and growth of Desulfovibrio on the surface of agar plates in an atmosphere of hydrogen. Sodium lactate does not appear to be essential in this medium for good growth and sulfate reduction in a hydrogen atmosphere, but is essential in a nitrogen atmosphere. Growth of Desulfovibrio (hydrogen atmosphere) on the agar surface of media commonly used for its cultivation as well as on an inorganic medium containing bicarbonate as a source of carbon is poor and erratic unless inoculated (Desulfovibrio) plates of TSA plus salts are incubated in the same container with plates of these media. This stimulatory effect of incubation with inoculated plates of TSA plus salts medium appears to be due to as yet unidentified volatile material produced by D. desulfuricans when growing on this medium. Another volatile material, or possibly the identical material, appears to act similarly to a hydrogen donor. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:5955798

  13. Influence of volume expansion on NaC1 reabsorption in the diluting segments of the nephron: a study using clearance methods.

    PubMed

    Danovitch, G M; Bricker, N S

    1976-09-01

    Whether volume expansion influences NaC1 reabsorption by the diluting segment of the nephron remains a matter of controversy. In the present studies this question has been examined in normal unanesthetized dogs, undergoing maximal water diuresis. Free water clearance (CH2O/GFR) has been used as the index of NaC1 reabsorption in the diluting segment. Three expressions have been employed for "distal delivery" of NaC1: a) V/GFR, designated as the "volume term"; b) (CNa/GFR + CH2O/GFR), the "sodium term;" and c) (CC1/GFR + CH2O/GFR), the "chloride term". The validity of these terms is discussed. Three techniques were used to increase distal delivery: 1) the administration of acetazolamide to dogs in which extracellular fluid (ECF) volume was not expanded (grop 1); 2) "moderate" volume expansion (group 2); and 3) "marked" volume expansion (group 3). CH2O/GFR increased progressively with rising values for "distal delivery" regardless of which term was used to calculate the latter. With all three delivery terms, differences in distal NaC1 reabsorption emerged between the two volume-expanded groups, though only with the "chloride" term did substantial differences also emerge between the nonexpanded group 1 dogs and both volume-expanded groups. In group 1, values for CH2O/GFR increased in close to a linear fashion up to distal delivery values equal to 24% of the volume of glomerular filtrate. However, at high rates of distal delivery the rate of rise of CH2O/GFR was less in group 2 than in group 1 and the depression of values was even greater in group 3. Within the limits of the techniques used, the data suggest that volume expansion inhibits fractional NaC1 reabsorption in the diluting segment of the nephron in a dose-related fashion. The "chloride" term was found to be superior to the "volume" and "sodium" terms in revealing these changes.

  14. Diluted magnetic semiconductors

    NASA Astrophysics Data System (ADS)

    Anderson, James R.

    1990-03-01

    Growth and physical properties of diluted magnetic semiconductors (DMS) were investigated. Growth included Bridgman, solid state recrystallization, and liquid phase epitaxy of Mercury(1-x)Manganese(x)Telluride and Mercury(1-x-y)Manganese(x)Cadmium(y)Telluride. Very uniform crystals were produced by solid state recrystallization. Physical properties studied included magnetization, optical response, and magnetotransport. From magnetization, the exchange interactions among magnetic ions have been deduced. Modulated spectroscopy gave details of the electronic structure of DMS and the quality of the material was indicated by the line widths. Magnetotransport, carried out in some cases to 30 T, showed a large negative magnetoresistance and subsequent increase. The Hg(1-x-y)Mn(x)Cd(y)Te has considerable promise for avalanche photodiodes between 1.2 and 1.8 micrometers.

  15. Dilution jet mixing program

    NASA Technical Reports Server (NTRS)

    Srinivasan, R.; Coleman, E.; Johnson, K.

    1984-01-01

    Parametric tests were conducted to quantify the mixing of opposed rows of jets (two-sided injection) in a confined cross flow. Results show that jet penetrations for two sided injections are less than that for single-sided injections, but the jet spreading rates are faster for a given momentum ratio and orifice plate. Flow area convergence generally enhances mixing. Mixing characteristics with asymmetric and symmetric convergence are similar. For constant momentum ratio, the optimum S/H(0) with in-line injections is one half the optimum value for single sided injections. For staggered injections, the optimum S/H(0) is twice the optimum value for single-sided injection. The correlations developed predicted the temperature distributions within first order accuracy and provide a useful tool for predicting jet trajectory and temperature profiles in the dilution zone with two-sided injections.

  16. Is blood agar an alternative to sabouraud dextrose agar for the isolation of fungi in patients with mycotic keratitis.

    PubMed

    Reddy, Ashok Kumar; Brahmaiah, Upputuri; Narayen, Nitesh; Reddy, Ravi Kumar; Reddy, Rupak Kumar; Chitta, Meghraj; Prasad, Srinivas; Swarup, Rishi; Mohiuddin, Syed Maaz; Reddy, Madhukar; Aasuri, Murali K; Murthy, B S R; Bhide, Milind; Ahmed, Sajid

    2013-06-01

    To compare the blood agar (BA), sabouraud dextrose agar (SDA) and chocolate agar (CA) for the isolation of fungi in patients with mycotic keratitis. Corneal Scrapings of 229 patients with clinically diagnosed microbial keratitis were inoculated on BA, SDA, CA. The culture media were evaluated for the rate and time taken for the fungal growth. Seventy six of 229 patients had fungal keratitis. Fungus grew on BA in 60/76(78.9 %), on SDA in 76/76 (100 %), on CA in 40/76(52.6 %) patients. The fungi which grew on BA (60/76) also grown on SDA at the same time. The colony morphologies of different fungi were better on SDA than BA/CA. Among the different culture media, SDA is essential for the isolation fungi in patients with mycotic keratitis.

  17. Diluted magnetic semiconductor nanowires exhibiting magnetoresistance

    DOEpatents

    Yang, Peidong; Choi, Heonjin; Lee, Sangkwon; He, Rongrui; Zhang, Yanfeng; Kuykendal, Tevye; Pauzauskie, Peter

    2011-08-23

    A method for is disclosed for fabricating diluted magnetic semiconductor (DMS) nanowires by providing a catalyst-coated substrate and subjecting at least a portion of the substrate to a semiconductor, and dopant via chloride-based vapor transport to synthesize the nanowires. Using this novel chloride-based chemical vapor transport process, single crystalline diluted magnetic semiconductor nanowires Ga.sub.1-xMn.sub.xN (x=0.07) were synthesized. The nanowires, which have diameters of .about.10 nm to 100 nm and lengths of up to tens of micrometers, show ferromagnetism with Curie temperature above room temperature, and magnetoresistance up to 250 Kelvin.

  18. [Methicillin resistance detection in Staphylococcus aureus: comparison between conventional methods and MRSA-Screen latex agglutination technique].

    PubMed

    Soloaga, R; Corso, A; Gagetti, P; Faccone, D; Galas, M

    2004-01-01

    Methicillin-resistant Staphylococcus aureus (MRSA) is a significant pathogen that has emerged over the last four decades, causing both nosocomial and community-acquired infections. Rapid and accurate detection of methicillin resistance in S. aureus is important for the use of appropriate antimicrobial therapy and for the control of nosocomial spread of MRSA strains. We evaluated the efficiency of conventional methods for detection of methicillin resistance such as the disk diffusion, agar dilution, oxacillin agar screen test, and the latex agglutination test MRSA-Screen latex, in 100 isolates of S. aureus, 79 mecA positive and 21 mecA negative. The MRSA-Screen latex (Denka Seiken, Niigata, Japón), is a latex agglutination method that detects the presence of PLP-2a, product of mecA gene in S. aureus. The PCR of the mecA gene was used as the "gold standard" for the evaluation of the different methods tested. The percentages of sensitivity and specificity were as follows: disk difusión 97 and 100%, agar dilution 97 and 95%, oxacillin agar screen test 100 and 100%, and MRSA-Screen latex, 100 and 100 %. All methods presented high sensitivity and specificity, but MRSA-Screen latex had the advantage of giving a reliable result, equivalent to PCR, in only 15 minutes.

  19. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  20. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  1. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  2. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  3. 21 CFR 866.4600 - Ouchterlony agar plate.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Ouchterlony agar plate. 866.4600 Section 866.4600 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunology Laboratory Equipment and Reagents §...

  4. Improving agar electrospinnability with choline-based deep eutectic solvents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One percent agar (% wt) was dissolved in the deep eutectic solvent (DES), (2-hydroxyethyl) trimethylammonium chloride/urea at a 1:2 molar ratio, and successfully electrospun into nanofibers. An existing electrospinning set-up, operated at 50 deg C, was adapted for use with an ethanol bath to collect...

  5. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion–reaction based continuum model

    NASA Astrophysics Data System (ADS)

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-08-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion–reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm’s shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here.

  6. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion-reaction based continuum model.

    PubMed

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-01-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion-reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm's shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here. PMID:27434099

  7. Simulation of Bacillus subtilis biofilm growth on agar plate by diffusion-reaction based continuum model

    NASA Astrophysics Data System (ADS)

    Zhang, Xianlong; Wang, Xiaoling; Nie, Kai; Li, Mingpeng; Sun, Qingping

    2016-08-01

    Various species of bacteria form highly organized spatially-structured aggregates known as biofilms. To understand how microenvironments impact biofilm growth dynamics, we propose a diffusion-reaction continuum model to simulate the formation of Bacillus subtilis biofilm on an agar plate. The extended finite element method combined with level set method are employed to perform the simulation, numerical results show the quantitative relationship between colony morphologies and nutrient depletion over time. Considering that the production of polysaccharide in wild-type cells may enhance biofilm spreading on the agar plate, we inoculate mutant colony incapable of producing polysaccharide to verify our results. Predictions of the glutamate source biofilm’s shape parameters agree with the experimental mutant colony better than that of glycerol source biofilm, suggesting that glutamate is rate limiting nutrient for Bacillus subtilis biofilm growth on agar plate, and the diffusion-limited is a better description to the experiment. In addition, we find that the diffusion time scale is of the same magnitude as growth process, and the common-employed quasi-steady approximation is not applicable here.

  8. Dilute acid/metal salt hydrolysis of lignocellulosics

    DOEpatents

    Nguyen, Quang A.; Tucker, Melvin P.

    2002-01-01

    A modified dilute acid method of hydrolyzing the cellulose and hemicellulose in lignocellulosic material under conditions to obtain higher overall fermentable sugar yields than is obtainable using dilute acid alone, comprising: impregnating a lignocellulosic feedstock with a mixture of an amount of aqueous solution of a dilute acid catalyst and a metal salt catalyst sufficient to provide higher overall fermentable sugar yields than is obtainable when hydrolyzing with dilute acid alone; loading the impregnated lignocellulosic feedstock into a reactor and heating for a sufficient period of time to hydrolyze substantially all of the hemicellulose and greater than 45% of the cellulose to water soluble sugars; and recovering the water soluble sugars.

  9. Stress in dilute suspensions

    NASA Technical Reports Server (NTRS)

    Passman, Stephen L.

    1989-01-01

    Generally, two types of theory are used to describe the field equations for suspensions. The so-called postulated equations are based on the kinetic theory of mixtures, which logically should give reasonable equations for solutions. The basis for the use of such theory for suspensions is tenuous, though it at least gives a logical path for mathematical arguments. It has the disadvantage that it leads to a system of equations which is underdetermined, in a sense that can be made precise. On the other hand, the so-called averaging theory starts with a determined system, but the very process of averaging renders the resulting system underdetermined. A third type of theory is proposed in which the kinetic theory of gases is used to motivate continuum equations for the suspended particles. This entails an interpretation of the stress in the particles that is different from the usual one. Classical theory is used to describe the motion of the suspending medium. The result is a determined system for a dilute suspension. Extension of the theory to more concentrated systems is discussed.

  10. Infrared thermography analysis of thermal diffusion induced by RF magnetic field on agar phantoms loaded with magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Bante-Guerra, Jose; Macías, J. D.; Caballero-Aguilar, L.; Vales-Pinzón, C.; Alvarado-Gil, J. J.

    2013-02-01

    Recently, several treatments for fighting malignant tumors have been designed. However these procedures have well known inconveniences, depending on their applicability, tumor size and side effects, among others. Magnetic hyperthermia is a safe, non-invasive method for cancer therapy. This treatment is applied via elevation of target tissue temperature by dissipation of heat from Magnetic Nanoparticles (MNPs), previously located within the tumor. The induction of heat causes cell death and therefore the removal of the tumor. In this work the thermal diffusion in phantoms of agar loaded with magnetic nanoparticles (MNPs) is studied using the infrared thermography technique, which is widely used in biology/medicine (e.g. skin temperature mapping). Agar is one of the materials used to simulate different types of body tissues, these samples are known as "phantoms". Agar is of natural origin, low cost and high degree of biocompatibility. In this work the agar gel was embedded with MNPs by coprecipitation and placed in an alternating magnetic field radiation. As a consequence, the energy from the radiation source is dissipated as heat and then transferred from the MNP to the gel, increasing its temperature. For the temperature analysis, the samples of agar gel were stimulated by RF magnetic field generated by coils. Heating was measured with infrared thermography using a Thermovision A20M infrared camera. Thermographic images allowed obtaining the dependence of thermal diffusion in the phantom as a function of the magnitude of the applied RF magnetic field and the load of magnetic particles.

  11. Preservation of anthocyanins in solid lipid nanoparticles: Optimization of a microemulsion dilution method using the Placket-Burman and Box-Behnken designs.

    PubMed

    Ravanfar, Raheleh; Tamaddon, Ali Mohammad; Niakousari, Mehrdad; Moein, Mahmoud Reza

    2016-05-15

    Anthocyanins are the main polyphenol components from red cabbage (Brassica oleracea L. Var. Capitata f. Rubra) extracts that have inherent antioxidant activities. Anthocyanins are effectively stable in acidic gastric digestion conditions, with nearly 100% phenol content recovery. However, the total phenol content recovery after simulated pancreatic digestion was approximately 25%. To protect anthocyanins against harsh environmental conditions (e.g., pH and temperature), solid lipid nanoparticles were prepared by the dilution of water in oil (w/o) microemulsions containing anthocyanins in aqueous media. The formulations were characterized for particle size and encapsulation efficiency. The formulation parameters (e.g., volume of the internal aqueous phase, homogenization time and the percentages of total lipid, total surfactant or stabilizer) were optimized using the Placket-Burman and Box-Behnken experimental designs. Entrapment efficiency (89.2 ± 0.3%) was calculated when the mean particle size was 455 ± 2 nm. A scanning electron microscopy study revealed the spherical morphology of the particles. PMID:26776010

  12. Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method

    PubMed Central

    1993-01-01

    B-less mice express a human immunoglobulin (Ig) lambda transgene that induces a severe deficiency of both immature pre-B and mature B lymphocytes. To understand this perturbation in B lymphopoiesis, we have devised a sensitive limiting dilution polymerase chain reaction assay that quantitates specific Ig rearrangements and thus quantitates B lineage cells at various stages of differentiation within unfractionated bone marrow. We find that there are significantly reduced frequencies of both VH-to-DJH and VK-to-JK rearrangements in the transgenic strain, whereas the frequency of D-to-JH rearrangements approximates that of wild type. Since Ig gene rearrangements occur in a stepwise fashion in which D-to-JH joining precedes that of VH-to-DJH and VK-to-JK, these results indicate that the major block of B lymphocyte development in the B-less strain occurs after D-to-JH rearrangement. Interestingly, sequence analysis of residual VHDJH junctions from transgenic pre-B lymphocytes reveals that an abnormally high proportion of these are out of frame and therefore nonproductive. Taken together, these data suggest that early expression of the transgenic lambda protein specifically prevents the development of a normal-sized population of precursor B lymphocytes coexpressing functional IgH. The transgene-induced immunodeficiency appears to arise by a precocious maturation process in which precursors bypass a developmental stage associated with cellular expansion. PMID:8315387

  13. Development of an equilibrium headspace gas chromatographic method for the measurement of noncovalent association and partitioning of n-alkylbenzenes at infinite dilution in fulvic acid pseudophase.

    PubMed

    Eljack, Mahmoud D; Wilson, Rachael E; Hussam, Abul; Khan, Shahamat U

    2015-02-27

    Fulvic acid (FA), the most important water soluble fraction of humic substances in nature, is known to form aggregate pseudophase and complexes with organic and inorganic species. Here, we report a novel equilibrium headspace gas chromatography (eHSGC) and a two-step reaction model to measure n-alkylbenzene-FA association constant (K11) and n-alkylbenzene-pseudophase FAn association constant (Kn1) without solute concentration and response factor. The K11 and Kn1 values were 2-3 orders of magnitude higher than those for sodium dodecylsulfate. Changes in peak area were used to calculate the critical FA-aggregation concentration (cfc), mole fraction based partition coefficients (Kx), activity coefficients of solute inside the aggregate pseudophase (γm(∞)), and transfer free energies of alkyl CH2 at infinite dilution. The cfc was found to be 10±0.5μM. The Kx values are of the order of 10(7) in the FA-aggregate pseudophase. The data shows that benzene has the lowest (0.0002) and n-butylbenzene has the highest (0.01) γm(∞) values, which are seven orders of magnitude smaller than γw(∞) in water. The transfer free energy of association of a CH2 group, -155cal/mol, compared to that of benzene, -9722cal/mol, indicates that the FA-aggregate pseudophase is more polarizable benzene-like and less n-alkane aliphatic-like.

  14. Agar-agar entrapment increases the stability of endo-β-1,4-xylanase for repeated biodegradation of xylan.

    PubMed

    Bibi, Zainab; Shahid, Faiza; Ul Qader, Shah Ali; Aman, Afsheen

    2015-04-01

    Microbial xylanases, specially endo-β-1,4-xylanase catalyzes the hydrolysis of xylan, is considered one of the most significant hydrolases. It has numerous applications but most extensively is utilized in paper and pulp industry as a bio-bleaching agent. Immobilization technique is comprehensively studied with the expectation of modifying and improving enzyme stability and characteristics for commercial purposes. Currently, matrix entrapment technique is applied to immobilize endo-β-1,4-xylanase within agar-agar gel beads produced by Geobacillus stearothermophilus KIBGE-IB29. Maximal enzyme immobilization yield was achieved at 2.5% of agar-agar concentration. Optimized conditions demonstrated an increase in the optimal reaction time from 05 min to 30 min and incubation temperature from 50 °C to 60 °C with reference to free enzyme whereas; no effect was observed for optimum pH. Entrapment technique uniquely changed the kinetic parameters of immobilized endo-β-1,4-xylanase (Km: 0.5074 mg min(-1) to 0.5230 mg min(-1) and Vmax: 4773 U min(-1) to 968 U min(-1)) as compared to free enzyme. However, immobilized enzyme displayed broad thermal stability and retained 79.0% of its initial activity at 80 °C up to 30 min whereas; free enzyme completely lost its activity at this temperature. With respect to economic feasibility, the immobilized enzyme showed impressive recycling efficiency up to six reaction cycles. PMID:25603143

  15. Thermal-induced ageing of agar solutions: impact on the structural and mechanical properties of agar gels

    NASA Astrophysics Data System (ADS)

    Mao, Bosi; Bentaleb, Ahmed; Louerat, Frédéric; Divoux, Thibaut; Snabre, Patrick

    Numerous hydrogels are prepared by cooling down to ambient temperature, aqueous polymer solutions brought to a boil. Although the incubation time of the polymer solution at such a high temperature could be used as a tuning parameter, its impact on the subsequent gelation has been poorly investigated. Here we study the effect of prolonged heating at 80°C on a 1.5% wt solution of agar, a natural polysaccharide. The incubation time is varied from a few hours up to five days. We show that the agar sol. continuously degrades as the result of both the hydrolysis and the intermolecular oxidation of the polymer chains. Furthermore, electronic microscopy and X-ray diffraction experiments reveal that gels formed from older agar sols display an increasingly coarser microstructure composed of micron-sized aggregated pieces of polysaccharides, in contrast with the fibrous-like structure of gels made from fresh sols. Along with structural changes prolonged incubation time leads to weaker gels of lower shear elastic modulus. Finally, macro-indentation experiments coupled to direct visualization show that increasing the incubation time of the agar sol. decreases the yield strain of the gel by a factor of three, while the rupture scenario turns continuously from brittle to ductile-like. Acknowledging funding from BioMérieux & CNRS.

  16. Qualitative and quantitative agar invasion test based on bacterial colony/biofilm.

    PubMed

    Corcuera, María Teresa; Gómez-Aguado, Fernando; Gómez-Lus, María Luisa; Ramos, Carmen; de la Parte, María Antonia; Alonso, María José; Prieto, José

    2013-09-01

    Invasion of the culture medium is a feature frequently studied in yeasts, in which it has been related to a greater virulence, but it is practically unknown in bacteria. Recently, it has been demonstrated that several clinically relevant bacterial species were also able of invading agar media, so it was necessary to design a microbiological assay to study the expression of this character in bacteria. Accordingly, a bacterial agar invasion test based on colony/biofilm development was designed, which allows qualitative and quantitative characterization of bacterial growth into the agar culture medium. Once the culture conditions were optimized, the test was applied to 90 strains from nine bacterial species, validating its usefulness for differentiating invasive strains (positive) from those non invasive (negative). The test also allows sorting invasive strains according to agar invasion intensity (low, moderate, high) and topographic invasion pattern (peripheral, homogeneous, mixed). Moreover, an image analysis routine to quantify the invasion was developed. Implemented method enables direct measuring of two invasion parameters (invasion area and number of invasion dots), automated calculation of three relative variables (invasion relative area, invasion dots relative density, and invasion dot average area), and the establishment of strain specific frequency histograms. This new methodology is simple, fast, reproducible, objective, inexpensive and can be used to study a great number of specimens simultaneously, all of which make it suitable for incorporation to the routine of any microbiology laboratory. It could also be a useful tool for additional studies related to clinical aspects of bacterial isolates such as virulence and antimicrobial response.

  17. Evaluation of three decarboxylating agar media to detect histamine and tyramine-producing bacteria in ripened sausages.

    PubMed

    Roig-Sagués, A X; Hernàndez-Herrero, M M; López-Sabater, E I; Rodríguez-Jerez, J J; Mora-Ventura, M T

    1997-11-01

    Histidine- and tyrosine-decarboxylase activity of 175 strains of bacteria isolated from eight retail samples of Spanish ripened sausages was tested in three decarboxylating agars (Niven medium, Joosten and Northolt medium and modified decarboxylating agar of Maijala) and confirmed by an enzymic method (histamine) and thin-layer chromatography (tyramine). Enterobacteria and pseudomonads showed the highest percentage of positive responses to histamine and tyramine in the three decarboxylating agars, but only enterobacteria were subsequently confirmed as histamine-producing. Confirmed tyramine-producing strains were all identified as enterococci or lactic acid bacteria. The medium described by Joosten and Northolt was more sensitive and faster at detecting tyramine-producing microorganisms. However, all three media failed to detect one histamine-positive strain of lactic acid bacteria used as a control.

  18. [Evaluation of the Epsilometer (Etest) method for the detection of tetracycline susceptibility in Paenibacillus larvae, the causal agent of American foulbrood disease of honeybees].

    PubMed

    Alippi, Adriana M; Reynaldi, Francisco J; López, Ana C

    2013-01-01

    American foulbrood (AFB) is a bacterial disease caused by the spore-forming, grampositive bacterium Paenibacillus larvae, which affects honeybee broods worldwide. The aim of this work was to compare the Epsilometer test (Etest) to the agar dilution method for testing a collection of 22 P. larvae strains to tetracycline by using MYPGP and Iso- Sensitest agars. Results showed that a categorical agreement of 100% was found when using Iso-Sensitest, while a categorical agreement of 86.36% was found (with 3 minor errors) when MYPGP was tested. In conclusion, the Etest could be a rapid and reliable method for testing MIC values of tetracycline in P. larvae only when used in combination with Iso-Sensitest agar. Nevertheless, these results should be confirmed with future studies involving a larger number of isolates.

  19. [Evaluation of the Epsilometer (Etest) method for the detection of tetracycline susceptibility in Paenibacillus larvae, the causal agent of American foulbrood disease of honeybees].

    PubMed

    Alippi, Adriana M; Reynaldi, Francisco J; López, Ana C

    2013-01-01

    American foulbrood (AFB) is a bacterial disease caused by the spore-forming, grampositive bacterium Paenibacillus larvae, which affects honeybee broods worldwide. The aim of this work was to compare the Epsilometer test (Etest) to the agar dilution method for testing a collection of 22 P. larvae strains to tetracycline by using MYPGP and Iso- Sensitest agars. Results showed that a categorical agreement of 100% was found when using Iso-Sensitest, while a categorical agreement of 86.36% was found (with 3 minor errors) when MYPGP was tested. In conclusion, the Etest could be a rapid and reliable method for testing MIC values of tetracycline in P. larvae only when used in combination with Iso-Sensitest agar. Nevertheless, these results should be confirmed with future studies involving a larger number of isolates. PMID:24401780

  20. How do microorganisms influence trace element uptake by plants? Screening in an agar model rhizosphere.

    NASA Astrophysics Data System (ADS)

    Marchetti, M.; Robinson, B. H.; Evangelou, M. W. H.; Vachey, A.; Schwitzguebel, J. P.; Bernier-Latmani, R.; Schulin, R.

    2009-04-01

    Trace elements (TE) are essential for humans and plants, but they may be toxic if their concentration is too high. For this reason, the management of TE in soils is very important. In some cases it may be necessary to increase the uptake of nutrients or TE by plants, for example in a biofortification perspective. Conversely, in some other cases TE uptake by plants should be decreased, for instance to avoid heavy metals entering the food chain via edible crops. Microorganisms living in the rhizosphere affect trace element (TE) uptake by plants. However, due to the complexity of this space and the variety of microorganisms that occur there, it is difficult to isolate the effect of any particular strain. To overcome this hurdle, we developed a system in which we grew plants under sterile conditions in agar and inoculated their rhizosphere with a single, well-defined microbial strain. For many years, agar has been used as a growth substrate for microorganisms and plant tissues. It is cheap, easy to use, and can be autoclaved to ensure its sterility. Because of its widespread use, an experiment conducted using this substrate can be reproduced under the same conditions in any laboratory. In contrast to soil, there is little interaction between the trace elements and the agar matrix. There are many studies investigating the influence of microorganisms on TE uptake by plants. However, so far only a small variety of microorganisms has been tested on few plant species. Therefore, the first objective of our research was to develop a method to rapidly screen a large variety of microorganisms on various plant species. Once this goal was achieved, we sought to study the effect of single, well-defined microbial strains on TE uptake by sunflower and wheat. The substrate for plants growth was a 10% agar solution prepared with modified Hoagland's solution and a TE solution containing 1 mg/kg Pb and molar equivalents of Cu, Ni and Zn. The agar solution was autoclaved and poured into

  1. Dilution refrigeration for space applications

    NASA Technical Reports Server (NTRS)

    Israelsson, U. E.; Petrac, D.

    1990-01-01

    Dilution refrigerators are presently used routinely in ground based applications where temperatures below 0.3 K are required. The operation of a conventional dilution refrigerator depends critically on the presence of gravity. To operate a dilution refrigerator in space many technical difficulties must be overcome. Some of the anticipated difficulties are identified in this paper and possible solutions are described. A single cycle refrigerator is described conceptually that uses forces other than gravity to function and the stringent constraints imposed on the design by requiring the refrigerator to function on the earth without using gravity are elaborated upon.

  2. Identification of cardiovascular dilution systems by contrast ultrasound.

    PubMed

    Mischi, Massimo; Jansen, Annemieke H M; Korsten, Hendrikus H M

    2007-03-01

    Indicator dilution techniques permit accurate measurements of important cardiovascular parameters, such as pulmonary blood volume (PBV) and ejection fraction (EF). However, their use is limited by the need for central catheterization. Contrast ultrasonography allows overcoming this problem. PBV and EF can be measured by a dilution system identification algorithm after detection of multiple dilution curves by an ultrasound scanner. In this paper, we present a system identification method that exploits the a priori knowledge on the dilution system and finds the optimum parameters for the parametric model representing the dilution system impulse response. No subsequent model interpolation is needed. Volume measurements show accurate in-vitro results and clinical feasibility, while 50 EF measurements in patients show a 0.88 correlation coefficient with echocardiographic biplane estimates. In conclusion, adding a priori knowledge to the system identification algorithm leads to increased accuracy and robustness of the method for PBV and EF measurements.

  3. Campylobacter in broiler slaughter samples assessed by direct count on mCCDA and Campy-Cefex agar.

    PubMed

    Gonsalves, Camila Cristina; Borsoi, Anderlise; Perdoncini, Gustavo; Rodrigues, Laura Beatriz; do Nascimento, Vladimir Pinheiro

    2016-01-01

    Campylobacter spp. cause foodborne illnesses in humans primarily through the consumption of contaminated chicken. The aim of this study was to evaluate the United States Department of Agriculture's (USDA) recommended methodology, protocol MLG 41.02, for the isolation, identification and direct plate counting of Campylobacter jejuni and C. coli samples from the broiler slaughtering process. A plating method using both mCCDA and Campy-Cefex agars is recommended to recover Campylobacter cells. It is also possible to use this method in different matrices (cloacal swabs and water samples). Cloacal swabs, samples from pre-chiller and post-chiller carcasses and samples of pre-chiller, chiller and direct supply water were collected each week for four weeks from the same flock at a slaughterhouse located in an abattoir in southern Brazil. Samples were analyzed to directly count Campylobacter spp., and the results showed a high frequency of Campylobacter spp. on Campy-Cefex agar. For the isolated species, 72% were identified as Campylobacter jejuni and 38% as Campylobacter coli. It was possible to count Campylobacter jejuni and Campylobacter coli from different samples, including the water supply samples, using the two-agar method. These results suggest that slaughterhouses can use direct counting methods with both agars and different matrices as a monitoring tool to assess the presence of Campylobacter bacteria in their products. PMID:27237112

  4. Mupirocin-mucin agar for selective enumeration of Bifidobacterium bifidum.

    PubMed

    Pechar, Radko; Rada, Vojtech; Parafati, Lucia; Musilova, Sarka; Bunesova, Vera; Vlkova, Eva; Killer, Jiri; Mrazek, Jakub; Kmet, Vladimir; Svejstil, Roman

    2014-11-17

    Bifidobacterium bifidum is a bacterial species exclusively found in the human intestinal tract. This species is becoming increasingly popular as a probiotic organism added to lyophilized products. In this study, porcine mucin was used as the sole carbon source for the selective enumeration of B. bifidum in probiotic food additives. Thirty-six bifidobacterial strains were cultivated in broth with mucin. Only 13 strains of B. bifidum utilized the mucin to produce acids. B. bifidum was selectively enumerated in eight probiotic food supplements using agar (MM agar) containing mupirocin (100 mg/L) and mucin (20 g/L) as the sole carbon source. MM agar was fully selective if the B. bifidum species was presented together with Bifidobacterium animalis subsp. lactis, Bifidobacterium breve, and Bifidobacterium longum subsp. longum species and with lactic acid bacteria (lactobacilli, streptococci). Isolated strains of B. bifidum were identified using biochemical, PCR, MALDI-TOF procedures and 16S rRNA gene sequencing. The novel selective medium was also suitable for the isolation of B. bifidum strains from human fecal samples.

  5. Modeling development of inhibition zones in an agar diffusion bioassay

    PubMed Central

    Chandrasekar, Vaishnavi; Knabel, Stephen J; Anantheswaran, Ramaswamy C

    2015-01-01

    A two-temperature agar diffusion bioassay is commonly used to quantify the concentration of nisin using Micrococcus luteus as the indicator microorganism. A finite element computational model based on Fick's second law of diffusion was used to predict the radius of the inhibition zone in this diffusion bioassay. The model developed was used to calculate nisin concentration profiles as a function of time and position within the agar. The minimum inhibitory concentration (MIC) of nisin against M. luteus was determined experimentally. The critical time (Tc) for growth of M. luteus within the agar diffusion bioassay was experimentally determined using incubation studies with nisin. The radius of the inhibition zone was predicted from the computational model as the location where the predicted nisin concentration at Tc was equal to MIC. The MIC was experimentally determined to be 0.156 μg mL−1, and Tc was determined to be 7 h. Good agreement (R2 = 0.984) was obtained between model-predicted and experimentally determined inhibition zone radii. PMID:26405525

  6. Mupirocin-mucin agar for selective enumeration of Bifidobacterium bifidum.

    PubMed

    Pechar, Radko; Rada, Vojtech; Parafati, Lucia; Musilova, Sarka; Bunesova, Vera; Vlkova, Eva; Killer, Jiri; Mrazek, Jakub; Kmet, Vladimir; Svejstil, Roman

    2014-11-17

    Bifidobacterium bifidum is a bacterial species exclusively found in the human intestinal tract. This species is becoming increasingly popular as a probiotic organism added to lyophilized products. In this study, porcine mucin was used as the sole carbon source for the selective enumeration of B. bifidum in probiotic food additives. Thirty-six bifidobacterial strains were cultivated in broth with mucin. Only 13 strains of B. bifidum utilized the mucin to produce acids. B. bifidum was selectively enumerated in eight probiotic food supplements using agar (MM agar) containing mupirocin (100 mg/L) and mucin (20 g/L) as the sole carbon source. MM agar was fully selective if the B. bifidum species was presented together with Bifidobacterium animalis subsp. lactis, Bifidobacterium breve, and Bifidobacterium longum subsp. longum species and with lactic acid bacteria (lactobacilli, streptococci). Isolated strains of B. bifidum were identified using biochemical, PCR, MALDI-TOF procedures and 16S rRNA gene sequencing. The novel selective medium was also suitable for the isolation of B. bifidum strains from human fecal samples. PMID:25217723

  7. Modeling development of inhibition zones in an agar diffusion bioassay.

    PubMed

    Chandrasekar, Vaishnavi; Knabel, Stephen J; Anantheswaran, Ramaswamy C

    2015-09-01

    A two-temperature agar diffusion bioassay is commonly used to quantify the concentration of nisin using Micrococcus luteus as the indicator microorganism. A finite element computational model based on Fick's second law of diffusion was used to predict the radius of the inhibition zone in this diffusion bioassay. The model developed was used to calculate nisin concentration profiles as a function of time and position within the agar. The minimum inhibitory concentration (MIC) of nisin against M. luteus was determined experimentally. The critical time (T c) for growth of M. luteus within the agar diffusion bioassay was experimentally determined using incubation studies with nisin. The radius of the inhibition zone was predicted from the computational model as the location where the predicted nisin concentration at T c was equal to MIC. The MIC was experimentally determined to be 0.156 μg mL(-1), and T c was determined to be 7 h. Good agreement (R (2) = 0.984) was obtained between model-predicted and experimentally determined inhibition zone radii.

  8. Impaction onto a Glass Slide or Agar versus Impingement into a Liquid for the Collection and Recovery of Airborne Microorganisms

    PubMed Central

    Juozaitis, Arvydas; Willeke, Klaus; Grinshpun, Sergey A.; Donnelly, Jean

    1994-01-01

    To study impaction versus impingement for the collection and recovery of viable airborne microorganisms, three new bioaerosol samplers have been designed and built. They differ from each other by the medium onto which the bioaerosol particles are collected (glass, agar, and liquid) but have the same inlet and collection geometries and the same sampling flow rate. The bioaerosol concentrations recorded by three different collection techniques have been compared with each other: impaction onto a glass slide, impaction onto an agar medium, and impingement into a liquid. It was found that the particle collection efficiency of agar slide impaction depends on the concentration of agar in the collection medium and on the sampling time, when samples are collected on a nonmoving agar slide. Impingement into a liquid showed anomalous behavior with respect to the sampling flow rate. Optimal sampling conditions in which all three new samplers exhibit the same overall sampling efficiency for nonbiological particles have been established. Inlet and collection efficiencies of about 100% have been achieved for all three devices at a sampling flow rate of 10 liters/min. The new agar slide impactor and the new impinger were then used to study the biological factors affecting the overall sampling efficiency. Laboratory experiments on the total recovery of a typical environmental microorganism, Pseudomonas fluorescens ATCC 13525, showed that both sampling methods, impaction and impingement, provided essentially the same total recovery when relatively nonstressed microorganisms were sampled under optimal sampling conditions. Comparison tests of the newly developed bioaerosol samplers with those commercially available showed that the incorporation of our research findings into the design of the new samplers yields better performance data than data from currently available samplers. PMID:16349217

  9. Intermittency in dilute granular flows

    NASA Astrophysics Data System (ADS)

    Guo, Wenxuan; Zhang, Qiang; Wylie, Jonathan J.

    2016-07-01

    In this letter, we show that dilute granular systems can exhibit a type of intermittency that has no analogue in gas dynamics. We consider a simple system in which a very dilute set of granular particles falls under gravity through a nozzle. This setting is analogous to the classical problem of high-speed nozzle flow in the study of compressible gases. It is well known that very dilute granular systems exhibit behavior qualitatively similar to gases, and that gas flowing through a nozzle does not exhibit intermittency. Nevertheless, we show that the intermittency in dilute granular nozzle flows can occur and corresponds to complicated transitions between supersonic and subsonic regimes. We also provide detailed explanations of the mechanism underlying this phenomenon.

  10. X-Plate Technology: a new method for detecting fluconazole resistance in Candida species.

    PubMed

    Chadwick, Sean G; Schuyler, Jessica A; Vermitsky, John-Paul; Adelson, Martin E; Mordechai, Eli; Gygax, Scott E

    2013-05-01

    Candida species are responsible for many opportunistic fungal infections. Fluconazole is a well-tolerated antifungal drug, commonly used in the treatment of candidiasis. However, with fluconazole resistance ever increasing, rapid detection and antifungal susceptibility testing of Candida is imperative for proper patient treatment. This paper reports a cost-effective, simple and rapid chromogenic agar dilution method for simultaneous Candida species identification and fluconazole susceptibility testing. The results obtained by X-Plate Technology were in absolute concordance with standard microbroth dilution assays. Analysis of 1383 clinical patient samples with suspected vulvovaginal candidiasis revealed that this technology was able to detect and speciate the Candida isolate and determine the fluconazole susceptibility. The prevalence and susceptibility profiles of the clinical isolates using this method were highly similar to published reports using the microbroth dilution method.

  11. 40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... dilution system, you may use a laminar flow element, an ultrasonic flow meter, a subsonic venturi, a... § 1065.240 Dilution air and diluted exhaust flow meters. (a) Application. Use a diluted exhaust flow meter to determine instantaneous diluted exhaust flow rates or total diluted exhaust flow over a...

  12. DAVINCI: Dilute Aperture VIsible Nulling Coronagraphic Imager

    NASA Technical Reports Server (NTRS)

    Shao, Michael; Levine, B. M.; Vasisht, G.; Lane, B. F.; Woodruff, R.; Vasudevan, G.; Samuele, R.; Lloyd, C. A.; Clampin, M.; Lyon, R.; Guyon, O.

    2008-01-01

    This slide presentation gives an overview of DAVINCI (Dilute Aperture VIsible Nulling Coronagraphic Imager). The presentation also includes information about dilute aperture coronagraph, and lyot efficiency.

  13. Differential recovery of Streptococcus mutans from various mitis-salivarius agar preparations.

    PubMed Central

    Liljemark, W F; Okrent, D H; Bloomquist, C G

    1976-01-01

    Recoveries of Streptococcus mutans from human dental plaque were lower when plated on mitis-salivarius agar obtained from Baltimore Biological Laboratories as compared with mitis-salivarius agar obtained from Difco Laboratories. However, no difference in recoveries of established laboratory strains of S. mutans was observed between these two agar preparations. PMID:956358

  14. Electrospinning of agar/PVA aqueous solutions and its relation with rheological properties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this work, we report the successful fabrication of agar-based nanofibers by an electrospinning technique using water as the solvent media. A tubeless spinneret was attached inside the electrospinning chamber, operated at 50 deg C, to avoid agar gelation. Pure agar solution 1% (w/w) showed inadequ...

  15. [Evaluation of the ChromID ESBL agar for the detection of ESBL-positive Enterobacteriaceae and vancomycin-resistant enterococcus isolates from urine cultures].

    PubMed

    Alışkan, Hikmet Eda; Colakoğlu, Sule; Turunç, Tuba; Demiroğlu, Yusuf Ziya

    2012-01-01

    Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae strains are frequent causative agents both in community-acquired infections and in nosocomial infections. The newly developed ChromID ESBL agar (bioMerieux, Marcy I'Etoile, France) is a chromogenic medium that helps rapid identification of ESBL-positive Enterobacteriaceae species from the clinical samples. The aim of this study was to evaluate the performance of ChromID ESBL agar in the rapid identification of ESBL-positive pathogens from the urine samples of the patients with urinary tract infections. A total of 672 urine samples (437 outpatients, 235 inpatients) were included in the study. All of the samples were inoculated simultaneously to 5% sheep blood agar, McConkey agar and ChromID ESBL agar media, and evaluated after incubation at 37°C for 18-24 hours. Gram-negative pathogens were tested for ESBL both by the standard combined double-disk diffusion (CDD) method using ceftazidime and cefotaxime disks and by doubledisk synergy (DDS) test. Among 672 urine cultures, 199 yielded microbial growth in routine media (sheep blood agar and/or McConkey agar), whereas 57 yielded bacterial growth in ChromID ESBL agar. When CDD method was accepted as the reference method according to Clinical and Laboratory Standards Institute (CLSI) recommendations, the sensitivity, specificity, positive and negative predictive values for ChromID ESBL agar for the detection of ESBL-positive bacteria in urinary tract infections were estimated as 97%, 92.9%, 89.1%, and 98.1%, respectively. Additionally, we also discovered that Chrom ID ESBL agar could detect vancomycin-resistant enterococci (VRE) as well as ESBL-positive bacteria, in our study. In order to investigate this observation we inoculated a total of 203 stock strains of Enterococcus spp. (118 vancomycin-sensitive, 85 vancomycin-resistant) to this medium. None of the vancomycinsensitive Enterococcus spp. did grow in ChromID ESBL medium, while 83 of the 85

  16. Multi-chamber electroosmosis using textile reinforced agar membranes--A promising concept for the future of hemodialysis.

    PubMed

    Kofler, Markus; Lenninger, Margit; Mayer, Gert; Neuwirt, Hannes; Grimm, Michael; Bechtold, Thomas

    2016-01-20

    Renal replacement therapy options are limited to hemodialysis and peritoneal dialysis (70% of US patients) or renal transplantation. Diffusion processes are the main physico-chemical principle behind hemodialysis. An alternative way to achieve liquid flow through membranes bases on the electroosmotic flow which is observed as electrokinetic phenomenon in porous membranes which bear surface charges. Agar consists of the non-ionic agarose and the negatively charged agaropectine thus an electroosmotic flux is observed in analytical electrophoresis. In this study the potential electroosmosis on textile reinforced agar membranes as separation method was investigated. Using a five-chamber electrolysis cell and an agar membrane/cellulose fabric composite an intensive electroosmotic flow of 1-2 ml cm(2) h(-1) at 100 mA cell current could be observed. The movement of cations in the negatively charged agar structure led to an intensive electroosmotic flux, which also transported uncharged molecules such as urea, glucose through the membrane. Separation of uncharged low molecular weight molecules is determined by the membrane characteristic. The transport of ions (K(+), PO4(3-), creatinine) and uncharged molecules (urea, glucose) in electroosmotic separation experiments was monitored using a pH 5.5 phosphate electrolyte with the aim to assess the overall transport processes in the electrochemical cell. The results demonstrate the potential of the method for filtration of biological fluids in the absence of external pressure or high shear rates.

  17. Cell-on-hydrogel platform made of agar and alginate for rapid, low-cost, multidimensional test of antimicrobial susceptibility.

    PubMed

    Sun, Han; Liu, Zhengzhi; Hu, Chong; Ren, Kangning

    2016-08-01

    Antimicrobial resistance (AMR) is a rapidly increasing threat to the effective treatment of infectious diseases worldwide. The two major remedies include: (1) using narrow-spectrum antibiotics based on rapid diagnosis; and (2) developing new antibiotics. A key part of both remedies is the antimicrobial susceptibility test (AST). However, the current standard ASTs that monitor colony formation are costly and time-consuming and the new strategies proposed are not yet practical to be implemented. Herein, we report a strategy to fabricate whole-hydrogel microfluidic chips using alginate-doped agar. This agar-based microfabrication makes it possible to prepare inexpensive hydrogel devices, and allows a seamless link between microfluidics and conventional agar-based cell culture. Different from common microfluidic systems, in our system the cells are cultured on top of the device, similar to normal agar plate culture; on the other hand, the microfluidic channels inside the hydrogel allow precise generation of linear gradient of drugs, thus giving a better performance than the conventional disk diffusion method. Cells in this system are not exposed to any shear flow, which allows the reliable tracking of individual cells and AST results to be obtained within 2-3 hours. Furthermore, our system could test the synergistic effect of drugs through two-dimensional gradient generation. Finally, the platform could be directly implemented to new drug discovery and other applications wherein a fast, cost-efficient method for studying the response of microorganisms upon drug administration is desirable. PMID:27452345

  18. Multi-chamber electroosmosis using textile reinforced agar membranes--A promising concept for the future of hemodialysis.

    PubMed

    Kofler, Markus; Lenninger, Margit; Mayer, Gert; Neuwirt, Hannes; Grimm, Michael; Bechtold, Thomas

    2016-01-20

    Renal replacement therapy options are limited to hemodialysis and peritoneal dialysis (70% of US patients) or renal transplantation. Diffusion processes are the main physico-chemical principle behind hemodialysis. An alternative way to achieve liquid flow through membranes bases on the electroosmotic flow which is observed as electrokinetic phenomenon in porous membranes which bear surface charges. Agar consists of the non-ionic agarose and the negatively charged agaropectine thus an electroosmotic flux is observed in analytical electrophoresis. In this study the potential electroosmosis on textile reinforced agar membranes as separation method was investigated. Using a five-chamber electrolysis cell and an agar membrane/cellulose fabric composite an intensive electroosmotic flow of 1-2 ml cm(2) h(-1) at 100 mA cell current could be observed. The movement of cations in the negatively charged agar structure led to an intensive electroosmotic flux, which also transported uncharged molecules such as urea, glucose through the membrane. Separation of uncharged low molecular weight molecules is determined by the membrane characteristic. The transport of ions (K(+), PO4(3-), creatinine) and uncharged molecules (urea, glucose) in electroosmotic separation experiments was monitored using a pH 5.5 phosphate electrolyte with the aim to assess the overall transport processes in the electrochemical cell. The results demonstrate the potential of the method for filtration of biological fluids in the absence of external pressure or high shear rates. PMID:26572331

  19. Bicosomes: Bicelles in Dilute Systems

    PubMed Central

    Rodríguez, Gelen; Soria, Guadalupe; Coll, Elisenda; Rubio, Laia; Barbosa-Barros, Lucyanna; López-Iglesias, Carmen; Planas, Anna M.; Estelrich, Joan; de la Maza, Alfons; López, Olga

    2010-01-01

    Abstract Bicelles are discoidal phospholipid nanostructures at high lipid concentrations. Under dilute conditions, bicelles become larger and adopt a variety of morphologies. This work proposes a strategy to preserve the discoidal morphology of bicelles in environments with high water content. Bicelles were formed in concentrated conditions and subsequently encapsulated in liposomes. Later dilution of these new structures, called bicosomes, demonstrated that lipid vesicles were able to isolate and protect bicelles entrapped inside them from the medium. Characterization of systems before and after dilution by dynamic light-scattering spectroscopy and cryo-transmission electron microscopy showed that free bicelles changed in size and morphology, whereas encapsulated bicelles remained unaltered by the effect of dilution. Free and entrapped bicelles (containing the paramagnetic contrast agent gadodiamide) were injected into rat brain lateral ventricles. Coronal and sagittal visualization was performed by magnetic resonance imaging. Whereas rats injected with free bicelles did not survive the surgery, those injected with bicosomes did, and a hyperintensity effect due to gadodiamide was observed in the cerebrospinal fluid. These results indicate that bicosomes are a good means of preserving the morphology of bicelles under dilution conditions. PMID:20643066

  20. Culture of Piscirickettsia salmonis on enriched blood agar.

    PubMed

    Mauel, Michael J; Ware, Cynthia; Smith, Pedro A

    2008-03-01

    Piscirickettsia salmonis is the etiologic agent of piscirickettsiosis, an economically significant disease of fish. Isolation of P. salmonis by culturing on fish cell lines has been the standard technique since the initial isolation of the organism. The ability to grow P. salmonis on artificial media would relieve facilities of the cost of maintaining cell lines, permit isolation at fish culture sites with fewer contamination problems, and allow easier transport of isolates to diagnostic facilities for confirmation assays. This report describes the successful culture of P. salmonis on enriched blood agar. PMID:18319435

  1. How do microorganisms influence trace element uptake by plants? Screening in an agar model rhizosphere.

    NASA Astrophysics Data System (ADS)

    Marchetti, M.; Robinson, B. H.; Evangelou, M. W. H.; Vachey, A.; Schwitzguebel, J. P.; Bernier-Latmani, R.; Schulin, R.

    2009-04-01

    Trace elements (TE) are essential for humans and plants, but they may be toxic if their concentration is too high. For this reason, the management of TE in soils is very important. In some cases it may be necessary to increase the uptake of nutrients or TE by plants, for example in a biofortification perspective. Conversely, in some other cases TE uptake by plants should be decreased, for instance to avoid heavy metals entering the food chain via edible crops. Microorganisms living in the rhizosphere affect trace element (TE) uptake by plants. However, due to the complexity of this space and the variety of microorganisms that occur there, it is difficult to isolate the effect of any particular strain. To overcome this hurdle, we developed a system in which we grew plants under sterile conditions in agar and inoculated their rhizosphere with a single, well-defined microbial strain. For many years, agar has been used as a growth substrate for microorganisms and plant tissues. It is cheap, easy to use, and can be autoclaved to ensure its sterility. Because of its widespread use, an experiment conducted using this substrate can be reproduced under the same conditions in any laboratory. In contrast to soil, there is little interaction between the trace elements and the agar matrix. There are many studies investigating the influence of microorganisms on TE uptake by plants. However, so far only a small variety of microorganisms has been tested on few plant species. Therefore, the first objective of our research was to develop a method to rapidly screen a large variety of microorganisms on various plant species. Once this goal was achieved, we sought to study the effect of single, well-defined microbial strains on TE uptake by sunflower and wheat. The substrate for plants growth was a 10% agar solution prepared with modified Hoagland's solution and a TE solution containing 1 mg/kg Pb and molar equivalents of Cu, Ni and Zn. The agar solution was autoclaved and poured into

  2. Standard Nutrient Agar 1 as a substitute for blood-supplemented Müller-Hinton agar for antibiograms in developing countries.

    PubMed

    Niederstebruch, N; Sixt, D

    2013-02-01

    In the industrial world, the agar diffusion test is a standard procedure for the susceptibility testing of bacteria isolates. Beta-hemolytic Streptococcus spp. are tested with Müller-Hinton agar supplemented with 5 % blood, a so-called blood agar. The results are interpreted using standardized tables, which only exist for this type of nutrient matrix. Because of a number difficulties, both with respect to technical issues and to manual skills, blood agar is not a feasible option in many developing countries. Beta-hemolytic Streptococcus spp. also grow on Standard Nutrient Agar 1 (StNA1). This suggests using that type of nutrient medium for running agar diffusion tests. However, there are no standardized tables that can be used for interpreting the diameters of the zones of inhibition on StNA1 1. Using the existing standardized tables for blood agar to interpret cultures on StNA1 1 would be of great benefit under such circumstances where blood agar is not available. With this in mind, we conducted comparative tests to evaluate the growth characteristics of beta-hemolytic Streptococcus spp. on StNA1 1 compared to Müller-Hinton agar supplemented with 5 % sheep blood. In this study, we were able to show that beta-hemolytic Streptococcus spp. develop similar zones of inhibition on blood agar and on StNA1 1. Therefore, it is suggested that, for the interpretation of antibiograms of beta-hemolytic Streptococcus spp. performed on StNA1 1, the standard tables for blood agar can be used.

  3. Standard Nutrient Agar 1 as a substitute for blood-supplemented Müller-Hinton agar for antibiograms in developing countries.

    PubMed

    Niederstebruch, N; Sixt, D

    2013-02-01

    In the industrial world, the agar diffusion test is a standard procedure for the susceptibility testing of bacteria isolates. Beta-hemolytic Streptococcus spp. are tested with Müller-Hinton agar supplemented with 5 % blood, a so-called blood agar. The results are interpreted using standardized tables, which only exist for this type of nutrient matrix. Because of a number difficulties, both with respect to technical issues and to manual skills, blood agar is not a feasible option in many developing countries. Beta-hemolytic Streptococcus spp. also grow on Standard Nutrient Agar 1 (StNA1). This suggests using that type of nutrient medium for running agar diffusion tests. However, there are no standardized tables that can be used for interpreting the diameters of the zones of inhibition on StNA1 1. Using the existing standardized tables for blood agar to interpret cultures on StNA1 1 would be of great benefit under such circumstances where blood agar is not available. With this in mind, we conducted comparative tests to evaluate the growth characteristics of beta-hemolytic Streptococcus spp. on StNA1 1 compared to Müller-Hinton agar supplemented with 5 % sheep blood. In this study, we were able to show that beta-hemolytic Streptococcus spp. develop similar zones of inhibition on blood agar and on StNA1 1. Therefore, it is suggested that, for the interpretation of antibiograms of beta-hemolytic Streptococcus spp. performed on StNA1 1, the standard tables for blood agar can be used. PMID:22926453

  4. Simple and sensitive analysis of histamine and tyramine in Japanese soy sauces and their intermediates using the stable isotope dilution HILIC-MS/MS method.

    PubMed

    Todoroki, Kenichiro; Ishii, Yasuhiro; Miyauchi, Chiemi; Kitagawa, Sachiyo; Min, Jun Zhe; Inoue, Koichi; Yamanaka, Tomoyuki; Suzuki, Kuniaki; Yoshikawa, Yuko; Ohashi, Norio; Toyo'oka, Toshimasa

    2014-07-01

    We established a simple, sensitive, and reproducible method to analyze the histamine and tyramine levels in Japanese soy sauce and its mash (called moromi) using hydrophilic interaction liquid chromatography with tandem mass spectrometry (HILIC-MS/MS). Histamine and tyramine quantification was performed using their stable isotopes for electrospray ionization-tandem mass spectrometry in the selected reaction monitoring mode. The sample pretreatment process was a simple, one-step liquid-liquid extraction. HILIC separation was performed with a gradient elution of aqueous ammonium formate and acetonitrile. Because of validation tests, the linearity, the accuracies, and precisions were sufficient. The limit of detection and the limit of quantification were 0.09 and 0.29 ppm for histamine and 0.13 and 0.42 ppm for tyramine, respectively. We successfully applied this method to histamine and tyramine determination in four kinds of commercial Japanese soy sauces and also in moromi samples during soy sauce production.

  5. Borelli's lactritmel agar induces conidiation in rare-macroconidia producing dermatophytic fungi.

    PubMed

    Ilkit, Macit; Gümral, Ramazan; Döğen, Aylin

    2012-10-01

    Macroconidia are among the most important indicators used to identify dermatophytic fungi, but several do not usually sporulate and/or produce macroconidia on Sabouraud glucose agar. Specifically, Microsporum audouinii, M. ferrugineum, Trichophyton concentricum, T. schoenleinii, T. verrucosum, and T. violaceum (including T. soudanense and T. yaoundei) rarely form macroconidia and, therefore, cannot be easily identified. In this study, we investigated the production of macroconidia on nine common laboratory media, including Borelli's lactritmel agar (BLA), modified Borelli's lactritmel agar (MBLA), brain heart infusion agar (BHIA), Christensen's urease agar in Petri dishes (UPA), cornmeal dextrose agar (CMDA), Lowenstein-Jensen agar (LJA), malt extract agar (MEA), oatmeal agar (OA), and potato dextrose agar (PDA). The performance of these media was evaluated using 18 rare-macroconidia producing isolates, including representative of the six species mentioned above. All cultures in this study were incubated at 26°C on the bench, and conidia formation on each was investigated at 5, 10, 15, 20, 25, and 30 days of incubation. BLA apparently improved macroconidia production after 15 days and was the most useful nutrient agar medium to induce these phenotypic characters in daily practice, closely followed by OA, PDA, and MBLA. PMID:22563856

  6. Physicochemical properties of biodegradable polyvinyl alcohol-agar films from the red algae Hydropuntia cornea.

    PubMed

    Madera-Santana, Tomás J; Robledo, Daniel; Freile-Pelegrín, Yolanda

    2011-08-01

    Agar obtained from the red alga Hydropuntia cornea was blended with polyvinyl alcohol (PVOH) in order to produce biodegradable films. In this study, we compare the properties of biopolymeric films formulated with agars extracted from H. cornea collected at different seasons (rainy and dry) in the Gulf of Mexico coast and PVOH as synthetic matrix. The films were prepared at different agar contents (0%, 25%, 50%, 75%, and 100%) and their optical, mechanical, thermal, and morphological properties analyzed. The tensile strength of PVOH-agar films increased when agar content was augmented. The formulation with 50% agar from rainy season (RS) had a significant higher tensile strength when compared to those from dry season (DS; p < 0.05). Tensile modulus also displayed an increasing trend and likewise, for 50% and 75% agar blends from RS showed higher values than those from DS (p < 0.05). In contrast, elongation at break decreased as the agar content increased, independently of the season. Environmental scanning electron microscopy images of PVOH-agar 75% biofilms from RS showed a homogeneous structure with good interfacial adhesion between the two components. The changes evidenced in the FTIR spectrum of this blend suggest that hydrogen bonding is taking place between the agar ether linkages (C-O-C) and the hydroxyl groups (OH) of the PVOH. Based on the above mentioned results, blends of PVOH and 75% agar from H. cornea collected in rainy season showed good properties for applications in the biodegradable packaging industry.

  7. [Clinical utility of Pourmedia GBS agar on screening for vaginal colonization of Group B Streptococcus].

    PubMed

    Kaneda, Mitsunori; Nagasaki, Hiromi; Tasaki, Megumi; Kamiyama, Kiyoshi

    2014-01-01

    Group B Streptococcus (GBS) are normal flora of the vagina and intestinal, but if the pregnant woman was infected with GBS in the vagina, miscarriage or premature would occur or the newborn would be developed to severe GBS infection. It is recommended that the inspection of GBS on all pregnant women by Japan Society of Obstetrics and Gynecology (JSOG) and Center for Disease Control and Prevention (CDC). We examined the comparison of detection rate between Pourmedia GBS agar (Eiken Chemical Co., Ltd.) and Nissui Separated Plate Sheep Blood Agar/BTB Lactose Agar medium (Nissui Pharmaceutical Co., Ltd.) on 112 sample. The positive rate of Pourmedia GBS agar was 21.4% (24/112 samples), Whereas Nissui Separated Plate Sheep Blood Agar/BTB Lactose Agar medium was 17.8% (20/112 samples). It was found that the detection rate was improved by using Pourmedia GBS agar on GBS screening test of vaginal swab.

  8. Usefulness of Chromogenic CromoCen® AGN agar medium for the identification of the genus Aeromonas: Assessment of faecal samples.

    PubMed

    Aguilera-Arreola, M G; Portillo-Muñoz, M I; Rodríguez-Martínez, C; Castro-Escarpulli, G

    2012-08-01

    Selective screening media for the detection and identification of Aeromonas strains are needed to guide primary isolation procedures in the clinical laboratory. This study compared the selective CromoCen® AGN chromogenic agar medium for the detection and identification of Aeromonas strains that were isolated from various samples against the conventional selective agar media that are commonly used for the isolation of this organism in food, environmental and clinical samples. The Miles and Misra and ecometric methods were used to evaluate the microbiological performance of CromoCen® AGN chromogenic agar medium, which was shown to be satisfactory. A total of 14 reference Aeromonas strains, 44 wild strains and 106 clinical stool specimens were examined using both non-chromogenic selective agars that are commonly used for Aeromonas isolation and CromoCen® AGN agar. The latter exhibited 94.73% sensitivity and 100% specificity for the various samples. On CromoCen® AGN agar medium, Aeromonas formed colonies with light green, greenish and salmon pigments with or without a surrounding wide transparent zone (halo) of 2-3mm in diameter around the entire border. This medium is recommended for the isolation and potential identification of the Aeromonas genus.

  9. Agar Medium for Differential Enumeration of Lactic Streptococci1

    PubMed Central

    Reddy, M. S.; Vedamuthu, E. R.; Washam, C. J.; Reinbold, G. W.

    1972-01-01

    An agar medium containing arginine and calcium citrate as specific substrates, diffusible (K2HPO4) and undiffusible (CaCO3) buffer systems, and bromocresol purple as the pH indicator was developed to differentiate among lactic streptococci in pure and mixed cultures. Milk was added as the sole source of carbohydrate (lactose) and to provide growth-stimulating factors. Production of acid from lactose caused developing bacterial colonies to seem yellow. Subsequent arginine utilization by Streptococcus lactis and S. diacetilactis liberated ammonia, resulting in a localized pH shift back toward neutrality and a return of the original purple indicator hue. The effects of production of acid from lactose and ammonia were fixed around individual colonies by the buffering capacity of CaCO3. After 36 hr at 32 C in a candle oats jar, colonies of S. cremoris were yellow, whereas colonies of S. lactis and S. diacetilactis were white. S. diacetilactis, on further incubation, utilized suspended calcium citrate, and, after 6 days, the citrate-degrading colonies exhibited clear zoning against a turbid background, making them easily distinguishable from the colonies of the other two species. The medium proved suitable for quantitative differential enumeration when compared with another widely used general agar medium for lactic streptococci. Images PMID:16349952

  10. Effect of dilution on carboxymethylcellulase and xylanase assays

    SciTech Connect

    Cauchon, N.; LeDuy, A.

    1984-08-01

    It has been demonstrated that the dilution of samples prior to the carboxymethylcellulase and xylanase assays causes serious discrepancies in the numerical values obtained for the enzyme activities. Even when the sample is assayed with the identical procedure, one could obtain different numerical values of the enzyme activity U depending on how much this sample has been diluted before the enzyme assay. Two crude commercial cellulase samples of Aspergillus niger and Trichoderma viride as well as the culture filtrate of our newly isolated acidophilic fungus have been used for the demonstration. An empirical method for reporting the cellulolytic activity by taking into account this dilution effect is proposed.

  11. An eco-friendly strategy, using on-line monitoring and dilution coupled to a second-order chemometric method, for the construction of dissolution curves of combined pharmaceutical associations.

    PubMed

    Calvo, Natalia L; Maggio, Rubén M; Kaufman, Teodoro S

    2014-02-01

    A simple, precise, economic and minimally operator-dependent method was developed under green analytical chemistry principles, for the simultaneous construction of the dissolution curves of a pharmaceutical association in short time and without employing organic solvents, allowing important savings of labor and resources. The carvedilol (CAR) and hydrochlorothiazide (HCT) combined formulation was employed as a model. The method (OD/UV-MCR) involves on-line sample dilution (OD) and UV detection of the analytes, coupled to multivariate curve resolution with alternating least squares (MCR-ALS). OD/UV-MCR proved to be robust and was successfully validated in accordance to ICH guidelines, fulfilling acceptance criteria for specificity (r(2) of spectral correlation>0.950), linearity [r>0.999 (N=25) in the ranges 1.00-31.1mg l(-1) and 0.51-15.2mg l(-1) for CAR and HCT, respectively] and precision (RSD<2%). Accuracy was assessed by point-to-point comparison between the dissolution profiles furnished by the proposed method with those provided by HPLC analysis, evidencing the usefulness of this monitoring system. In addition, OD/UV-MCR was successfully employed for the comparative analysis of three lots of commercial formulations of the CAR-HCT pharmaceutical association, belonging to a couple of different brands, employing Moore and Flanner's f2 similarity indicator. PMID:24291800

  12. Simultaneous Quantification of Methylated Cytidine and Adenosine in Cellular and Tissue RNA by Nano-Flow Liquid Chromatography-Tandem Mass Spectrometry Coupled with the Stable Isotope-dilution Method

    PubMed Central

    Fu, Lijuan; Amato, Nicolas J.; Wang, Pengcheng; McGowan, Sara J.; Niedernhofer, Laura J.; Wang, Yinsheng

    2016-01-01

    The rising interest in understanding the functions, regulation and maintenance of the epitranscriptome calls for robust and accurate analytical methods for the identification and quantification of post-transcriptionally modified nucleosides in RNA. Mono-methylations of cytidine and adenosine are common post-transcriptional modifications in RNA. Herein, we developed an LC-MS/MS/MS coupled with the stable isotope-dilution method for the sensitive and accurate quantifications of 5-methylcytidine (m5C), 2′-O-methylcytidine (Cm), N6-methyladenosine (m6A) and 2′-O-methyladenosine (Am) in RNA isolated from mammalian cells and tissues. Our results showed that the distributions of the four methylated nucleosides are tissue-specific. In addition, the 2′-O-methylated ribonucleosides (Cm and Am) are present at higher levels than the corresponding methylated nucleobase products (m5C and m6A) in total RNA isolated from mouse brain, pancreas and spleen, but not mouse heart. We also found that the levels of m5C, Cm and Am are significantly lower (by 6.5-43 fold) in mRNA than in total RNA isolated from HEK293T cells, whereas the level of m6A was slightly higher (by 1.6 fold) in mRNA than in total RNA. The availability of this analytical method, in combination with genetic manipulation, may facilitate the future discovery of proteins involved in the maintenance and regulation of these RNA modifications. PMID:26158405

  13. Development and validation of a new analytical method for the determination of 1,4-dichlorobenzene in honey by gas chromatography-isotope dilution mass spectrometry after steam-distillation.

    PubMed

    Botitsi, E V; Kormali, P N; Kontou, S N; Economou, A; Tsipi, D F

    2006-10-01

    A simple, fast, sensitive and robust analytical method using gas chromatography (GC)-isotope dilution mass spectrometry (MS) was developed and validated for the identification and quantification of 1,4-dichlorobenzene (p-DCB) residues in honey samples. The proposed methodology is based on steam-distillation using a Clevenger-type apparatus followed by gas chromatography-mass spectrometry (GC-MS) in the selected ion monitoring (SIM) mode employing the isotopically labeled analogue d4-1,4-dichlorobenzene (d4-p-DCB) as internal standard (IS). Validation of the method was performed in two different GC-MS systems, using quadrupole MS (QMS) and ion-trap MS (ITMS) detectors, with no statistically significant differences between two. Recoveries were better than 91% with percent relative standard deviations lower than 12%. The instrumental limits of detection were 1 microg kg(-1) in the GC-ITMS system and 0.6 microg kg(-1) in the GC-QMS system. The expanded uncertainty was estimated as 17% at the currently accepted "action level" of 10 microg kg(-1). The method was applied to the analysis of 310 honey samples in an extensive national monitoring study. A quality control (QC) system applied during the assays has demonstrated a good performance and long-term stability over a period of more than 8 months of continuous operation.

  14. An eco-friendly strategy, using on-line monitoring and dilution coupled to a second-order chemometric method, for the construction of dissolution curves of combined pharmaceutical associations.

    PubMed

    Calvo, Natalia L; Maggio, Rubén M; Kaufman, Teodoro S

    2014-02-01

    A simple, precise, economic and minimally operator-dependent method was developed under green analytical chemistry principles, for the simultaneous construction of the dissolution curves of a pharmaceutical association in short time and without employing organic solvents, allowing important savings of labor and resources. The carvedilol (CAR) and hydrochlorothiazide (HCT) combined formulation was employed as a model. The method (OD/UV-MCR) involves on-line sample dilution (OD) and UV detection of the analytes, coupled to multivariate curve resolution with alternating least squares (MCR-ALS). OD/UV-MCR proved to be robust and was successfully validated in accordance to ICH guidelines, fulfilling acceptance criteria for specificity (r(2) of spectral correlation>0.950), linearity [r>0.999 (N=25) in the ranges 1.00-31.1mg l(-1) and 0.51-15.2mg l(-1) for CAR and HCT, respectively] and precision (RSD<2%). Accuracy was assessed by point-to-point comparison between the dissolution profiles furnished by the proposed method with those provided by HPLC analysis, evidencing the usefulness of this monitoring system. In addition, OD/UV-MCR was successfully employed for the comparative analysis of three lots of commercial formulations of the CAR-HCT pharmaceutical association, belonging to a couple of different brands, employing Moore and Flanner's f2 similarity indicator.

  15. Development and validation of a new analytical method for the determination of 1,4-dichlorobenzene in honey by gas chromatography-isotope dilution mass spectrometry after steam-distillation.

    PubMed

    Botitsi, E V; Kormali, P N; Kontou, S N; Economou, A; Tsipi, D F

    2006-10-01

    A simple, fast, sensitive and robust analytical method using gas chromatography (GC)-isotope dilution mass spectrometry (MS) was developed and validated for the identification and quantification of 1,4-dichlorobenzene (p-DCB) residues in honey samples. The proposed methodology is based on steam-distillation using a Clevenger-type apparatus followed by gas chromatography-mass spectrometry (GC-MS) in the selected ion monitoring (SIM) mode employing the isotopically labeled analogue d4-1,4-dichlorobenzene (d4-p-DCB) as internal standard (IS). Validation of the method was performed in two different GC-MS systems, using quadrupole MS (QMS) and ion-trap MS (ITMS) detectors, with no statistically significant differences between two. Recoveries were better than 91% with percent relative standard deviations lower than 12%. The instrumental limits of detection were 1 microg kg(-1) in the GC-ITMS system and 0.6 microg kg(-1) in the GC-QMS system. The expanded uncertainty was estimated as 17% at the currently accepted "action level" of 10 microg kg(-1). The method was applied to the analysis of 310 honey samples in an extensive national monitoring study. A quality control (QC) system applied during the assays has demonstrated a good performance and long-term stability over a period of more than 8 months of continuous operation. PMID:17723727

  16. Automated on-line column-switching high performance liquid chromatography isotope dilution tandem mass spectrometry method for the quantification of bisphenol A, bisphenol F, bisphenol S, and 11 other phenols in urine.

    PubMed

    Zhou, Xiaoliu; Kramer, Joshua P; Calafat, Antonia M; Ye, Xiaoyun

    2014-01-01

    Human exposure to bisphenol A (BPA) is widespread. However, in recent years, bisphenol analogs such as bisphenol S (BPS) and bisphenol F (BPF) are replacing BPA in the production of some consumer products. Because human exposure to these alternative bisphenols may occur, biomonitoring of these bisphenol analogs is warranted. In the present study, we developed and validated a sensitive and selective method that uses on-line solid phase extraction coupled to high performance liquid chromatography-isotope dilution tandem mass spectrometry with peak focusing to measure BPA, BPF, BPS, and 11 other environmental phenols in urine. The method required a small amount of sample (100μL) and minimal sample pretreatment. The limits of detection were 0.03ng/mL (BPS), 0.06ng/mL (BPF), 0.10ng/mL (BPA), and ranged from 0.1ng/mL to 1.0ng/mL for the other 11 phenols. In 100 urine samples collected in 2009-2012 from a convenience group of anonymous adults in the United States, of the three bisphenols, we detected BPA at the highest frequency and median concentrations (95%, 0.72ng/mL), followed by BPS (78%, 0.13ng/mL) and BPF (55%, 0.08ng/mL). This sensitive, rugged, and labor and cost-effective method could be used for the analysis of large number of samples for epidemiologic studies.

  17. High-throughput assay of oleopentanedialdheydes in extra virgin olive oil by the UHPLC-ESI-MS/MS and isotope dilution methods.

    PubMed

    Di Donna, Leonardo; Benabdelkamel, Hicham; Mazzotti, Fabio; Napoli, Anna; Nardi, Monica; Sindona, Giovanni

    2011-03-15

    The quality of extra virgin olive oil is associated with the presence of microcomponents whose healing effects have been proved in some special cases. The enzymatic hydrolysis of oleuropein and ligstroside, and of their demethylated analogues, affords four different pentanedialdehydes, and for one of which, 2-(4-hydroxyphenyl)ethyl (3S,4E)-4-formyl-3-(2-oxoethyl)hex-4-enoate, also known as oleocanthal, an anti-inflammatory effect was quite recently carefully assessed. Extra virgin olive oil is now worldwide considered as a functional food whose daily intake, as for the Mediterranean diet, helps consumers in keeping a constant level of nonsteroidal anti-inflammatory drug (NSAID) in the blood. The presence of these active principles provides, therefore, olive oil with an important added value. In the framework of the actions of the recently funded Agrifood Regional Center, which should coordinate the scientific research and production worlds, an absolute analytical method was developed for the mass spectrometric detection of the two most abundant NSAIDs, Tyr-OLPD and HTyr-OLPD (oleopentanedialdehydes (OLPDs) conjugated to p-hydroxyphenylethanol and 3,4-dihydroxyphenylethanol, respectively), by UHPLC-ESI-MS/MS. PMID:21329394

  18. Dilution and the elusive baseline.

    PubMed

    Likens, Gene E; Buso, Donald C

    2012-04-17

    Knowledge of baseline conditions is critical for evaluating quantitatively the effect of human activities on environmental conditions, such as the impact of acid deposition. Efforts to restore ecosystems to prior, "pristine" condition require restoration targets, often based on some presumed or unknown baseline condition. Here, we show that rapid and relentless dilution of surface water chemistry is occurring in the White Mountains of New Hampshire, following decades of acid deposition. Extrapolating measured linear trends using a unique data set of up to 47 years, suggest that both precipitation and streamwater chemistry (r(2) >0.84 since 1985) in the Hubbard Brook Experimental Forest (HBEF) will approximate demineralized water within one to three decades. Because such dilute chemistry is unrealistic for surface waters, theoretical baseline compositions have been calculated for precipitation and streamwater: electrical conductivity of 3 and 5 μS/cm, base cation concentrations of 7 and 39 μeq/liter, acid-neutralizing capacity values of <1 and 14 μeq/liter, respectively; and pH 5.5 for both. Significantly large and rapid dilution of surface waters to values even more dilute than proposed for Pre-Industrial Revolution (PIR) conditions has important ecological, biogeochemical and water resource management implications, such as for the success of early reproductive stages of aquatic organisms.

  19. Dilution and the elusive baseline.

    PubMed

    Likens, Gene E; Buso, Donald C

    2012-04-17

    Knowledge of baseline conditions is critical for evaluating quantitatively the effect of human activities on environmental conditions, such as the impact of acid deposition. Efforts to restore ecosystems to prior, "pristine" condition require restoration targets, often based on some presumed or unknown baseline condition. Here, we show that rapid and relentless dilution of surface water chemistry is occurring in the White Mountains of New Hampshire, following decades of acid deposition. Extrapolating measured linear trends using a unique data set of up to 47 years, suggest that both precipitation and streamwater chemistry (r(2) >0.84 since 1985) in the Hubbard Brook Experimental Forest (HBEF) will approximate demineralized water within one to three decades. Because such dilute chemistry is unrealistic for surface waters, theoretical baseline compositions have been calculated for precipitation and streamwater: electrical conductivity of 3 and 5 μS/cm, base cation concentrations of 7 and 39 μeq/liter, acid-neutralizing capacity values of <1 and 14 μeq/liter, respectively; and pH 5.5 for both. Significantly large and rapid dilution of surface waters to values even more dilute than proposed for Pre-Industrial Revolution (PIR) conditions has important ecological, biogeochemical and water resource management implications, such as for the success of early reproductive stages of aquatic organisms. PMID:22455659

  20. Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test

    PubMed Central

    2010-01-01

    Background The ideal identification of Staphylococcus aureus clinical isolates requires a battery of tests and this is costly in resource limited settings. In many developing countries, the tube coagulase test is usually confirmatory for S. aureus and is routinely done using either human or sheep plasma. This study evaluated Mannitol salt agar and the deoxyribonuclease (DNase) test for improving the efficiency of the tube coagulase test in resource limited settings. The efficiency of human and sheep plasma with tube coagulase tests was also evaluated. Methods One hundred and eighty Gram positive, Catalase positive cocci occurring in pairs, short chains or clusters were subjected to growth on Mannitol salt agar, deoxyribonuclease and tube coagulase tests. Of these, isolates that were positive for at least two of the three tests (n = 60) were used to evaluate the performance of the tube coagulase test for identification of S. aureus, using PCR-amplification of the nuc gene as a gold standard. Results Human plasma was more sensitive than sheep plasma for the tube coagulase test (sensitivity of 91% vs. 81% respectively), but both plasmas had very low specificity (11% and 7% respectively). The sensitivity and specificity of the tube coagulase test (human plasma) was markedly improved when Mannitol salt agar and DNase were introduced as a tri-combination test for routine identification of Staphylococcus aureus (100% specificity and 75% sensitivity). The specificity and sensitivity of Mannitol salt agar/DNase/tube coagulase (sheep plasma) combination was 100% and 67%, respectively. Conclusion The efficiency of the tube coagulase test can be markedly improved by sequel testing of the isolates with Mannitol salt agar, DNase and Tube coagulase. There is no single phenotypic test (including tube coagulase) that can guarantee reliable results in the identification of Staphylococcus aureus. PMID:20707914

  1. Evaluation of eight agar media for the isolation of shiga toxin-Producing Escherichia coli.

    PubMed

    Gill, Alexander; Huszczynski, George; Gauthier, Martine; Blais, Burton

    2014-01-01

    The growth characteristics of 96 shiga toxin-producing Escherichia coli (STEC) strains representing 36 different O-types (including priority O types O26, O45, O103, O111, O121, O145 and O157) on commercial and in-house agar media were studied. The ability of the strains to grow on agar media with varying selective supplement formulations was evaluated using MacConkey Agar (MAC); Rainbow® Agar O157 (RBA); Rainbow® Agar O157 with manufacturer-recommended selective supplements (RBA-NT); Rainbow® Agar O157 with USDA-recommended selective supplements (RBA-USDA); CHROMagar STEC™ (CH STEC); Tryptone Bile agar containing cefixime and tellurite (TBA-CT); Tryptone Bile agar containing cefixime, tellurite, eosin and methylene blue (TBA-EM); and VTEC agar. All of the strains were able to grow on MAC, RBA and VTEC agar, whereas a number of strains (including some non-O157 priority O types) were unable to grow on the highly selective media CH STEC, RBA-NT, RBA-USDA, TBA-EM and TBA-CT. Only RBA-NT and CH STEC exhibited significant inhibition of background flora from ground beef enrichment. Significant inhibition of background flora from beef trim enrichment was observed with RBA-NT, RBA-USDA, CH STEC, TBA-EM and VTEC agar. With exception of E. coli O157, several different colony morphologies were observed on the differential plating media among strains of the same O type, indicating that this colony morphology is not a reliable means of identifying target STEC. These results suggest that an approach to maximize the recovery of target STEC from beef enrichment cultures is dual plating on lesser (RBA, MAC, VTEC agar) and more highly (RBA-NT, CH STEC) selective agars.

  2. How to optimize the drop plate method for enumerating bacteria.

    PubMed

    Herigstad, B; Hamilton, M; Heersink, J

    2001-03-01

    The drop plate (DP) method can be used to determine the number of viable suspended bacteria in a known beaker volume. The drop plate method has some advantages over the spread plate (SP) method. Less time and effort are required to dispense the drops onto an agar plate than to spread an equivalent total sample volume into the agar. By distributing the sample in drops, colony counting can be done faster and perhaps more accurately. Even though it has been present in the laboratory for many years, the drop plate method has not been standardized. Some technicians use 10-fold dilutions, others use twofold. Some technicians plate a total volume of 0.1 ml, others plate 0.2 ml. The optimal combination of such factors would be useful to know when performing the drop plate method. This investigation was conducted to determine (i) the standard deviation of the bacterial density estimate, (ii) the cost of performing the drop plate procedure, (iii) the optimal drop plate design, and (iv) the advantages of the drop plate method in comparison to the standard spread plate method. The optimal design is the combination of factor settings that achieves the smallest standard deviation for a fixed cost. Computer simulation techniques and regression analysis were used to express the standard deviation as a function of the beaker volume, dilution factor, and volume plated. The standard deviation expression is also applicable to the spread plate method.

  3. Characterization of Leptospiral Chemoreceptors Using a Microscopic Agar Drop Assay.

    PubMed

    Affroze, Samia; Islam, Md Shafiqul; Takabe, Kyosuke; Kudo, Seishi; Nakamura, Shuichi

    2016-08-01

    Bacterial chemotaxis is induced by sensing chemical stimuli via chemoreceptors embedded in the cytoplasmic membrane, enabling the cells to migrate toward nutrients or away from toxins. The chemoreceptors of Escherichia coli and Salmonella spp. have been well studied and are functionally classified on the basis of detectable substrates. The spirochete Leptospira possesses more than ten chemoreceptors and shows attractive or repellent responses against some sugars, amino acids, and fatty acids. However, the roles of these chemoreceptors have not been investigated. In this study, we conducted a chemotaxis assay called microscopic agar drop assay in combination with competition experiments, determining whether two kinds of attractants are recognized by the same type of chemoreceptor in the saprophytic Leptospira strain, Leptospira biflexa. Analyzing the competition effect observed between several pairs of chemicals, we found that L. biflexa senses sugars via chemoreceptors different from those that sense amino acids and fatty acids.

  4. Development, validation and application of a stable isotope dilution liquid chromatography electrospray ionization/selected reaction monitoring/mass spectrometry (SID-LC/ESI/SRM/MS) method for quantification of keto-androgens in human serum.

    PubMed

    Tamae, Daniel; Byrns, Michael; Marck, Brett; Mostaghel, Elahe A; Nelson, Peter S; Lange, Paul; Lin, Daniel; Taplin, Mary-Ellen; Balk, Steven; Ellis, William; True, Larry; Vessella, Robert; Montgomery, Bruce; Blair, Ian A; Penning, Trevor M

    2013-11-01

    Prostate cancer is the most frequently diagnosed form of cancer in males in the United States. The disease is androgen driven and the use of orchiectomy or chemical castration, known as androgen deprivation therapy (ADT) has been employed for the treatment of advanced prostate cancer for over 70 years. Agents such as GnRH agonists and non-steroidal androgen receptor antagonists are routinely used in the clinic, but eventually relapse occurs due to the emergence of castration-resistant prostate cancer. With the appreciation that androgen signaling still persists in these patients and the development of new therapies such as abiraterone and enzalutamide that further suppresses androgen synthesis or signaling, there is a renewed need for sensitive and specific methods to quantify androgen precursor and metabolite levels to assess drug efficacy. We describe the development, validation and application of a stable isotope dilution liquid chromatography electrospray ionization selected reaction monitoring mass spectrometry (SID-LC/ESI/SRM/MS) method for quantification of serum keto-androgens and their sulfate and glucuronide conjugates using Girard-T oxime derivatives. The method is robust down to 0.2-4pg on column, depending on the androgen metabolite quantified, and can also quantify dehydroepiandrosterone sulfate (DHEA-S) in only 1μL of serum. The clinical utility of this method was demonstrated by analyzing serum androgens from patients enrolled in a clinical trial assessing combinations of pharmacological agents to maximally suppress gonadal and adrenal androgens (Targeted Androgen Pathway Suppression, TAPS clinical trial). The method was validated by correlating the results obtained with a hydroxylamine derivatization procedure coupled with tandem mass spectrometry using selected reaction monitoring that was conducted in an independent laboratory.

  5. Detection of encapsulation in Staphylococcus aureus by use of antiserum agar.

    PubMed Central

    West, T E; Apicella, M A

    1984-01-01

    We examined an antiserum agar method to study its reliability in screening Staphylococcus aureus strains for capsule production. The encapsulated S. aureus Smith diffuse strain was compared with its nonencapsulated variant, Smith compact, in CCY medium containing 0.5% NaCl and 5.0% Smith diffuse rabbit antiserum. A halo was visible surrounding colonies of the Smith diffuse strain but not the Smith compact strain. On this same medium, the protein A-producing Cowan I strain possessed a halo that was visible on photographs. Single high-salt medium is known to inhibit protein A production, halo formation by the strains was also compared in 7.5% NaCl medium. The halo surrounding the Cowan I strain was not present when the salt content of the medium was increased. In contrast, the halo surrounding the Smith diffuse strain persisted in the 7.5% NaCl medium. By use of this medium, the antiserum agar technique may be valuable for the identification of encapsulated staphylococci without appreciable interference from protein A. Images PMID:6490810

  6. Visualization of Biosurfactant Film Flow in a Bacillus subtilis Swarm Colony on an Agar Plate.

    PubMed

    Kim, Kyunghoon; Kim, Jung Kyung

    2015-01-01

    Collective bacterial dynamics plays a crucial role in colony development. Although many research groups have studied the behavior of fluidic swarm colonies, the detailed mechanics of its motion remains elusive. Here, we developed a visualization method using submicron fluorescent beads for investigating the flow field in a thin layer of fluid that covers a Bacillus subtilis swarm colony growing on an agar plate. The beads were initially embedded in the agar plate and subsequently distributed spontaneously at the upper surface of the expanding colony. We conducted long-term live cell imaging of the B. subtilis colony using the fluorescent tracers, and obtained high-resolution velocity maps of microscale vortices in the swarm colony using particle image velocimetry. A distinct periodic fluctuation in the average speed and vorticity of flow in swarm colony was observed at the inner region of the colony, and correlated with the switch between bacterial swarming and growth phases. At the advancing edge of the colony, both the magnitudes of velocity and vorticity of flow in swarm colony were inversely correlated with the spreading speed of the swarm edge. The advanced imaging tool developed in this study would facilitate further understanding of the effect of micro vortices in swarm colony on the collective dynamics of bacteria. PMID:26343634

  7. Visualization of Biosurfactant Film Flow in a Bacillus subtilis Swarm Colony on an Agar Plate

    PubMed Central

    Kim, Kyunghoon; Kim, Jung Kyung

    2015-01-01

    Collective bacterial dynamics plays a crucial role in colony development. Although many research groups have studied the behavior of fluidic swarm colonies, the detailed mechanics of its motion remains elusive. Here, we developed a visualization method using submicron fluorescent beads for investigating the flow field in a thin layer of fluid that covers a Bacillus subtilis swarm colony growing on an agar plate. The beads were initially embedded in the agar plate and subsequently distributed spontaneously at the upper surface of the expanding colony. We conducted long-term live cell imaging of the B. subtilis colony using the fluorescent tracers, and obtained high-resolution velocity maps of microscale vortices in the swarm colony using particle image velocimetry. A distinct periodic fluctuation in the average speed and vorticity of flow in swarm colony was observed at the inner region of the colony, and correlated with the switch between bacterial swarming and growth phases. At the advancing edge of the colony, both the magnitudes of velocity and vorticity of flow in swarm colony were inversely correlated with the spreading speed of the swarm edge. The advanced imaging tool developed in this study would facilitate further understanding of the effect of micro vortices in swarm colony on the collective dynamics of bacteria. PMID:26343634

  8. Isolation of Shiga Toxin-Producing Escherichia coli from Ground Beef Using Multiple Combinations of Enrichment Broths and Selective Agars.

    PubMed

    Brusa, Victoria; Piñeyro, Pablo E; Galli, Lucía; Linares, Luciano H; Ortega, Emanuel E; Padola, Nora L; Leotta, Gerardo A

    2016-03-01

    Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens, and beef cattle are recognized as the principal reservoir. The aims of this study were (1) to identify the most sensitive combination of selective enrichment broths and agars for STEC isolation in artificially inoculated ground beef samples, and (2) to evaluate the most efficient combination(s) of methods for naturally contaminated ground beef samples. A total of 192 ground beef samples were artificially inoculated with STEC and non-stx bacterial strains. A combination of four enrichment broths and three agars were evaluated for sensitivity, specificity, and positive predictive value for STEC isolation from experimentally inoculated samples. Enrichments with either modified tryptic soy broth (mTSB) containing 8 mg/L novobiocin (mTSB-8) or modified Escherichia coli (mEC) broth followed by isolation in MacConkey agar were the most sensitive combinations for STEC isolation of artificially inoculated samples. Independently, both enrichments media followed by isolation in MacConkey were used to evaluate ground beef samples from 43 retail stores, yielding 65.1% and 58.1% stx-positive samples by RT-PCR, respectively. No difference was observed in the isolate proportions between these two methods (8/25 [32%] and 8/28 [28.6%]). Identical serotypes and stx genotypes were observed in STEC strains isolated from the same samples by either method. In this study, no single enrichment protocol was sufficient to detect all STEC in artificially inoculated samples and had considerable variation in detection ability with naturally contaminated samples. Moreover, none of the single or combinations of multiple isolation agars used were capable of identifying all STEC serogroups in either artificially inoculated or naturally occurring STEC-contaminated ground beef. Therefore, it may be prudent to conclude that there is no single method or combination of isolation methods capable of identifying all STEC serogroups

  9. SI-traceable certification of the amount content of cadium below the ng g(-1) level in blood samples by isotope dilution ICP-MS applied as a primary method of measurement.

    PubMed

    Diemer, J; Vogl, J; Quétel, C R; Linsinger, T; Taylor, P D; Lamberty, A; Pauwels, J

    2001-07-01

    The development and implementation of a method for the certification of cadmium in blood samples at low ng g(-1) and sub ng g(-1) levels is described. The analytical procedure is based on inductively coupled plasma isotope dilution mass spectrometry (ICP-IDMS) applied as a primary method of measurement. Two different sample digestion methods, an optimized microwave digestion procedure using HNO3 and H2O2 as oxidizing agents and a high-pressure asher digestion procedure, were developed and compared. The very high salt content of the digests and the high molybdenum content, which can cause oxide-based interferences with the Cd isotopes, were reduced by a chromatographic matrix separation step using an anion-exchange resin. All isotope ratio measurements were performed by a quadrupole ICP-MS equipped with an ultrasonic nebulizer with membrane desolvator. This sample introduction set-up was used to increase sensitivity and minimize the formation of oxides (less MoO+ interference with the Cd isotopes). Because of the very low Cd concentrations in the samples and the resulting need to minimize the procedural blank as much as possible, all sample-processing steps were performed in a clean room environment. Detection limits of 0.005 ng g(-1) Cd were achieved using sample weights of 2.7 g. The method described was used to recertify the cadmium content of three different blood reference materials from the Community Bureau of Reference (BCR) of the European Commission (BCR-194, BCR-195, BCR- 196). Cadmium concentrations ranged between approximately 0.2 ng g(-1) and approximately 12 ng g(-1). For these materials, SI-traceable certified values including total uncertainty budgets according to ISO and Eurachem guidelines were established.

  10. Quantification of viral proteins of the avian H7 subtype of influenza virus: an isotope dilution mass spectrometry method applicable for producing more rapid vaccines in the case of an influenza pandemic.

    PubMed

    Santana, Wanda I; Williams, Tracie L; Winne, Emily K; Pirkle, James L; Barr, John R

    2014-05-01

    Vaccination is the most effective means to prevent influenza and its serious complications. Influenza viral strains undergo rapid mutations of the surface proteins hemagglutinin (HA) and neuraminidase (NA) requiring vaccines to be frequently updated to include current circulating strains. It is nearly impossible to predict which strains will be circulating in the next influenza season. It is, therefore, imperative that the process of producing a vaccine be streamlined and as swift as possible. We have developed an isotope dilution mass spectrometry (IDMS) method to quantify HA and NA in H7N7, H7N2, and H7N9 influenza. The IDMS method involves enzymatic digestion of viral proteins and the specific detection of evolutionarily conserved target peptides. The four target peptides that were initially chosen for analysis of the HA protein of H7N2 and H7N7 subtypes were conserved and available for analysis of the H7N9 subtype that circulated in China in the spring of 2013. Thus, rapid response to the potential pandemic was realized. Quantification of a protein is performed by employing multiple peptides to ensure that the enzymatic digestion of the protein is efficient in the region of the target peptides, verify the accuracy of the measurement, and provide flexibility in the case of amino acid changes among newly emerging strains. The IDMS method is an accurate, sensitive, and selective method to quantify the amount of HA and NA antigens in primary liquid standards, crude allantoic fluid, purified virus samples, and final vaccine presentations.

  11. Broth and agar hop-gradient plates used to evaluate the beer-spoilage potential of Lactobacillus and Pediococcus isolates.

    PubMed

    Haakensen, M; Schubert, A; Ziola, B

    2009-03-15

    Identification of the beer-spoilage Lactobacillus and Pediococcus bacteria has largely taken two approaches; identification of spoilage-associated genes or identification of specific species of bacteria regardless of ability to grow in beer. The problem with these two approaches is that they are either overly inclusive (i.e., detect all bacteria of a given species regardless of spoilage potential) or overly selective (i.e., rely upon individual, putative spoilage-associated genes). Our goal was to design a method to assess the ability of Lactobacillus and Pediococcus to spoil beer that is independent of speciation or genetic background. In searching for a method by which to differentiate between beer-spoilage bacteria and bacteria that cannot grow in beer, we explored the ability of lactobacilli and pediococci isolates to grow in the presence of varying concentrations of hop-compounds and ethanol in broth medium versus on agar medium. The best method for differentiating between bacteria that can grow in beer and bacteria that do not pose a threat as beer-spoilage organisms was found to be a hop-gradient agar plate containing ethanol. This hop-gradient agar plate technique provides a rapid and simple solution to the dilemma of assessing the ability of Lactobacillus and Pediococcus isolates to grow in beer, and provides new insights into the different strategies used by these bacteria to survive under the stringent conditions of beer.

  12. Assessment of oxacillin salt agar for detection of MRSA identified by presence of the mecA gene.

    PubMed

    Kobayashi, Y; Kizaki, M; Kawakami, Y; Uchida, H; Ikeda, Y

    1993-04-01

    To assess the screening method for detection of methicillin-resistant Staphylococcus aureus recommended by Thornsberry and McDougal J. Clin Microbiol 1983; 18: 1084-1901, the growth of S. aureus and Staphylococcus epidermidis strains on Mueller Hinton agar containing 4% NaCl and 6 mg l-1 oxacillin, after 24 h incubation at 35 degrees C, was investigated. The strains used for this study were characterized for possession of the mecA gene by the polymerase chain reaction. All 39 strains of S. aureus with the mecA gene grew on this agar, and all 12 strains of S. aureus without the mecA gene did not grow. On the other hand, three of 12 strains of S. epidermidis did not grow on this agar, although all these strains possessed the mecA gene. These results suggest that Thornsberry and McDougal's screening method is suitable for detecting S. aureus strains with mecA gene. Introduction of this method, to detect methicillin-resistant S. aureus more precisely, is simple and cheap for any laboratory in any part of the world.

  13. Assessment of oxacillin salt agar for detection of MRSA identified by presence of the mecA gene.

    PubMed

    Kobayashi, Y; Kizaki, M; Kawakami, Y; Uchida, H; Ikeda, Y

    1993-04-01

    To assess the screening method for detection of methicillin-resistant Staphylococcus aureus recommended by Thornsberry and McDougal J. Clin Microbiol 1983; 18: 1084-1901, the growth of S. aureus and Staphylococcus epidermidis strains on Mueller Hinton agar containing 4% NaCl and 6 mg l-1 oxacillin, after 24 h incubation at 35 degrees C, was investigated. The strains used for this study were characterized for possession of the mecA gene by the polymerase chain reaction. All 39 strains of S. aureus with the mecA gene grew on this agar, and all 12 strains of S. aureus without the mecA gene did not grow. On the other hand, three of 12 strains of S. epidermidis did not grow on this agar, although all these strains possessed the mecA gene. These results suggest that Thornsberry and McDougal's screening method is suitable for detecting S. aureus strains with mecA gene. Introduction of this method, to detect methicillin-resistant S. aureus more precisely, is simple and cheap for any laboratory in any part of the world. PMID:8099927

  14. Effect of Different Commerical Agar Preparations on the Inhibitory Activities of Phenols

    PubMed Central

    Sands, J. G.; Bennett, E. O.

    1966-01-01

    The minimal inhibitory concentrations of 11 phenolic inhibitors were compared in five commercial agars and in nutrient broth. It was found that the brand of agar affected the end point obtained for a particular inhibitor, and that the degree of antagonism varied with each compound studied. The results indicate that there are at least two deleterious factors present in agar, one of which is water-soluble and one which is not. The major portion of the total antagonism was due to the water-soluble factor, which could be removed by washing the agar in warm distilled water prior to use in the test medium. PMID:5959856

  15. Detection of toxigenic isolates of Aspergillus flavus and related species on coconut cream agar.

    PubMed

    Dyer, S K; McCammon, S

    1994-01-01

    A new readily-prepared medium, coconut cream agar, was developed for the detection of aflatoxin production by isolates of Aspergillus flavus and related species. Coconut cream agar, which comprised coconut cream (50%) and agar (1.5%), detected isolates of A. flavus more effectively than the synthetic media tested and was as effective as media containing desiccated coconut. Fluorescence colouring of colonies grown on coconut cream agar could be used to differentiate A. flavus from A. parasiticus and A. nomius. In addition, conidial colour of A. flavus and A. nomius was quite distinct from that of A. parasiticus.

  16. Isolation of Shiga toxin-producing Escherichia coli from fresh produce using STEC heart infusion washed blood agar with mitomycin-C.

    PubMed

    Lin, Andrew; Nguyen, Lam; Clotilde, Laurie M; Kase, Julie A; Son, Insook; Lauzon, Carol R

    2012-11-01

    The ability to detect and isolate Shiga toxin-producing Escherichia coli (STEC) remains a major challenge for food microbiologists. Although methods based on nucleic acids and antibodies have improved detection of STECs in foods, isolation of these bacteria remains arduous. STEC isolation is necessary for matching food, environmental, and clinical isolates during outbreak investigations and for distinguishing between pathogenic and nonpathogenic organisms. STEC heart infusion washed blood agar with mitomycin-C (SHIBAM) is a modification of washed sheep blood agar prepared by adding mitomycin-C and optimizing both the washed blood and base agar to better isolate STECs. Most STEC isolates produce a zone of hemolysis on SHIBAM plates and are easily distinguishable from background microbiota. Here, we present data supporting the use of SHIBAM to isolate STECs from fresh produce. SHIBAM was tested for accuracy in identifying STECs (365 of 410 STEC strains were hemolytic, and 63 of 73 E. coli strains that did not produce Shiga toxin were not hemolytic) and for recovery from artificially inoculated fresh produce (11 of 24 romaine lettuce samples and 6 of 24 tomato samples). STEC recovery with SHIBAM agar was greatly improved when compared with recovery on Levine's eosin-methylene blue agar as a reference method.

  17. A rapid detection of multidrug-resistant Mycobacterium tuberculosis by a nitrate reductase assay on blood agar.

    PubMed

    Coban, Ahmet Yilmaz; Cayci, Yeliz Tanriverdi; Deveci, Aydin; Akgunes, Alper; Uzun, Meltem; Durupinar, Belma

    2011-05-01

    The susceptibility of 49 Mycobacterium tuberculosis clinical isolates to isoniazid (INH) and rifampisin (RIF) (28 multi-drug resistant-tuberculosis samples) was determined by a nitrate reductase assay (NRA) on blood agar. Agreement between the NRA and other testing methods was found to be 93.8% for both INH and RIF. The sensitivity, specificity, positive predictive value and negative predictive value for INH were 92.8%, 94.2%, 86.6% and 97%, respectively. The sensitivity, specificity, positive predictive value and negative predictive value for RIF were 90.4%, 96.4%, 95% and 93.1%. In conclusion, we show here that blood agar can be used effectively for the NRA test.

  18. Science Notes: Dilution of a Weak Acid

    ERIC Educational Resources Information Center

    Talbot, Christopher; Wai, Chooi Khee

    2014-01-01

    This "Science note" arose out of practical work involving the dilution of ethanoic acid, the measurement of the pH of the diluted solutions and calculation of the acid dissociation constant, K[subscript a], for each diluted solution. The students expected the calculated values of K[subscript a] to be constant but they found that the…

  19. Glucose-sucrose-potassium tellurite-bacitracin agar, an alternative to mitis salivarius-bacitracin agar for enumeration of Streptococcus mutans.

    PubMed Central

    Tanzer, J M; Börjesson, A C; Laskowski, L; Kurasz, A B; Testa, M

    1984-01-01

    An agar medium for selective recovery and enumeration of Streptococcus mutans was developed as an alternative to mitis salivarius-bacitracin (MSB) agar. Combinations of dyes, antibiotics, and tellurite were added to a nonselective medium which, because of its sucrose content, allowed easy recognition of S. mutans colonies. Candle jar incubation for 2 days, by comparison with anaerobic incubation, reduced background flora but did not diminish S. mutans recoveries from clinical samples. Quantitative comparisons were made of the simultaneous recoveries of a number of authentic S. mutans serotype representatives and fresh clinical isolates, using various glucose-sucrose-potassium tellurite-bacitracin (GSTB) formulations and mitis salivarius, MSB, and blood agars. Mitis salivarius counts were not detectably different from blood counts, but counts on MSB were distinctly lower. A formulation of the new medium containing 5% glucose 5% sucrose, 0.001% potassium tellurite, 0.3 U of bacitracin per ml (hence GSTB), and 2% agar gave recoveries nearly equal to those on mitis salivarius agar and much greater than those on MSB. The medium yielded readily recognized S. mutans colonies and facilitated detection of intracellular polysaccharide formers upon flooding with I2 reagent. Freshly isolated serotype c, E, and f colonies could often be distinguished from serotype d and g colonies, a distinction made reliable by testing for intracellular polysaccharide. A study of 300 salivary samples revealed GSTB to give significantly higher recoveries than MSB. About 72% of all samples were substantially underestimated for S. mutans with MSB, and 6.7% of samples were falsely negative for S. mutans with MSB. Recovery of background flora on GSTB was as low or lower than on MSB, and both types of agar could be stored for at least 9 weeks without notable change of selectivity. Thus, GSTB agar appears to be simple and reliable to use and requires no anaerobic incubation. Caution is voiced about

  20. Method to simultaneously determine the sphingosine 1-phosphate breakdown product (2E)-hexadecenal and its fatty acid derivatives using isotope-dilution HPLC-electrospray ionization-quadrupole/time-of-flight mass spectrometry.

    PubMed

    Neuber, Corinna; Schumacher, Fabian; Gulbins, Erich; Kleuser, Burkhard

    2014-09-16

    Sphingosine 1-phosphate (S1P), a bioactive lipid involved in various physiological processes, can be irreversibly degraded by the membrane-bound S1P lyase (S1PL) yielding (2E)-hexadecenal and phosphoethanolamine. It is discussed that (2E)-hexadecenal is further oxidized to (2E)-hexadecenoic acid by the long-chain fatty aldehyde dehydrogenase ALDH3A2 (also known as FALDH) prior to activation via coupling to coenzyme A (CoA). Inhibition or defects in these enzymes, S1PL or FALDH, result in severe immunological disorders or the Sjögren-Larsson syndrome, respectively. Hence, it is of enormous importance to simultaneously determine the S1P breakdown product (2E)-hexadecenal and its fatty acid metabolites in biological samples. However, no method is available so far. Here, we present a sensitive and selective isotope-dilution high performance liquid chromatography-electrospray ionization-quadrupole/time-of-flight mass spectrometry method for simultaneous quantification of (2E)-hexadecenal and its fatty acid metabolites following derivatization with 2-diphenylacetyl-1,3-indandione-1-hydrazone and 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide. Optimized conditions for sample derivatization, chromatographic separation, and MS/MS detection are presented as well as an extensive method validation. Finally, our method was successfully applied to biological samples. We found that (2E)-hexadecenal is almost quantitatively oxidized to (2E)-hexadecenoic acid, that is further activated as verified by cotreatment of HepG2 cell lysates with (2E)-hexadecenal and the acyl-CoA synthetase inhibitor triacsin C. Moreover, incubations of cell lysates with deuterated (2E)-hexadecenal revealed that no hexadecanoic acid is formed from the aldehyde. Thus, our method provides new insights into the sphingolipid metabolism and will be useful to investigate diseases known for abnormalities in long-chain fatty acid metabolism, e.g., the Sjögren-Larsson syndrome, in more detail. PMID:25137547

  1. Subculture on potato dextrose agar as a complement to the broth microdilution assay for Malassezia pachydermatis.

    PubMed

    Prado, Marilena R; Brito, Erika H S; Brilhante, Raimunda S N; Cordeiro, Rossana A; Leite, João J G; Sidrim, José J C; Rocha, Marcos F G

    2008-10-01

    The main aim of this study was to verify the efficacy of subculture on potato dextrose agar (PDA) as a complement to the in vitro susceptibility test for Malassezia pachydermatis strains by a broth microdilution method, as well as to determine the MIC and MFC of azole derivatives, amphotericin B and caspofungin. The microdilution assay was performed in 96-well plates using a modified RPMI 1640 medium. The M. pachydermatis strains were resistant to caspofungin. All strains (n=50) had shown MIC values of <0.03, <0.03, 2.0, 4.0 and 4.0 microg/ml for itraconazole, ketoconazole, voriconazole, fluconazole and amphotericin B, respectively. Thus, the subculture on PDA improved the analysis of the in vitro antifungal susceptibility of M. pachydermatis.

  2. Use of cefoperazone MacConkey agar for selective isolation of Laribacter hongkongensis.

    PubMed

    Lau, Susanna K P; Woo, Patrick C Y; Hui, Wai-ting; Li, Maria W S; Teng, Jade L L; Que, Tak-Lun; Luk, Wei-Kwang; Lai, Raymond W M; Yung, Raymond W H; Yuen, Kwok-yung

    2003-10-01

    A new selective medium, cefoperazone MacConkey agar (CMA), was developed for primary isolation of Laribacter hongkongensis from stool. Its performance in quantitative recovery and in a clinical evaluation of 4,741 human diarrheal stool specimens was superior to that of charcoal cefoperazone deoxycholate agar. In addition, with CMA, Arcobacter butzleri was unexpectedly isolated from the stools of six patients.

  3. Ceftibuten-containing agar plate for detecting group B streptococci with reduced penicillin susceptibility (PRGBS).

    PubMed

    Kamiya, Chitose; Kimura, Kouji; Doyama, Yo; Miyazaki, Akira; Morimoto, Makiko; Banno, Hirotsugu; Nagano, Noriyuki; Jin, Wanchun; Wachino, Jun-ichi; Yamada, Keiko; Arakawa, Yoshichika

    2015-08-01

    Penicillins remain first-line agents for treatment of group B Streptococcus (Streptococcus agalactiae; GBS) infections; however, several reports have confirmed the existence of GBS with reduced penicillin susceptibility (PRGBS). Because no selective agar plates for detection of PRGBS are available to date, in this investigation, we developed the selective agar plate for detection of PRGBS. We used 19 genetically well-confirmed PRGBS isolates and 38 penicillin-susceptible GBS isolates identified in Japan. For preparation of trial PRGBS-selective agar plates, we added 1 of antimicrobial agents (among oxacillin, ceftizoxime, and ceftibuten) to a well-established GBS-selective agar plate. Among 12 trial PRGBS-selective agar plates, Muller-Hinton agar containing 128 μg/mL ceftibuten with 5% sheep blood, 8 μg/mL gentamicin, and 12 μg/mL nalidixic acid was the most appropriate selective agar for PRGBS, showing 100% sensitivity and 81.6% specificity. In cases of potential nosocomial spread of PRGBS, the selective agar plate could be useful and reliable.

  4. Evaluation of a chromogenic agar for detection of group B streptococcus in pregnant women.

    PubMed

    Craven, Robin R; Weber, Carol J; Jennemann, Rebecca A; Dunne, W Michael

    2010-09-01

    We compared ChromID Strepto B agar (STRB; bioMérieux, Inc.), a selective and differential medium for group B streptococcus, with culture using neomycin-nalidixic acid agar (NNA) and LIM broth. STRB alone was more sensitive (87.7%) than NNA alone (79.0%), while each had a sensitivity of 100% when used in conjunction with LIM broth.

  5. High Concentrations of Manganese in Mueller-Hinton Agar Increase MICs of Tigecycline Determined by Etest▿

    PubMed Central

    Fernández-Mazarrasa, Carlos; Mazarrasa, Olav; Calvo, Jorge; del Arco, Asunción; Martínez-Martínez, Luis

    2009-01-01

    MICs of tigecycline determined by Etest were 4 to 12 times (three ATCC strains) and 2 to 8 times (50 clinical isolates) higher in Mueller-Hinton agar from Merck than in Mueller-Hinton agar from either Oxoid or Difco. This was related to a much higher concentration of manganese in the medium from Merck. PMID:19144806

  6. Effects of season on the yield and quality of agar from Gracilaria species (Gracilariaceae, Rhodophyta).

    PubMed

    Marinho-Soriano, E; Bourret, E

    2003-12-01

    The effect of season on yield and physical properties of agars extracted from Gracia gracilis and G. bursa-pastoris were determined. The agar yield from G. gracilis was maximum during spring (30%) and minimum during autumn (19%). In G. bursa-pastoris, the agar yield was greatest in summer (36%) and lowest in winter (23%). Agar yield from G. bursa-pastoris was positively correlated with temperature (r=0.94; P<0.01) and salinity (r=0.97; P<0.01) and negatively with nitrogen content (r=-0.93; P<0.01). Agar gel strengths fluctuated from 229 to 828 gcm(-2) and 23 to 168 gcm(-2) for G. gracilis and G. bursa-pastoris, respectively. The gelling temperature showed significant seasonal variation for both species. Chemical analysis of agar from the two seaweeds indicated variation in 3,6-anhydrogalactose and sulfate content (P<0.01). Furthermore, there was an inverse correlation between the two chemical variables. In general, agar extracted from G. gracilis possessed better qualities than agar extracted from G. bursa-pastoris and can be considered a candidate for industrial use.

  7. Development of novel agar media for isolating guaiacol producing Alicyclobacillus spp.

    PubMed

    Chang, S S; Park, S H; Kang, D H

    2013-06-01

    The purpose of this study is to develop a selective and differential medium (SK2 agar) for isolating guaiacol producing Alicyclobacillus. Forty-one selected dyes and vanillic acid were incorporated in SK agar for screening selective and differential agents. Two guaiacol producing (1016, 1101) and two non-guaiacol producing (19220, C-GD 1-1) Alicyclobacillus isolates were streaked onto media and color differentiation of the isolates was assessed. Among 41 tested dyes, Chrome Azurol S (CAS) allowed color differentiation of the two types of Alicyclobacillus. Colonies of guaiacol producing Alicyclobacillus isolates appeared as dark purple to royal blue color with yellow background, whereas non-guaiacol producing Alicyclobacillus isolates produced cream colored colonies with yellow background. Vanillic acid not only served as a precursor for guaiacol formation but also inhibited non-guaiacol producing Alicyclobacillus. Non-guaiacol producing isolates did not grow on SK agar containing more than 70 ppm vanillic acid, whereas the recovery of guaiacol producing isolates was unaffected. When compared with other Alicyclobacillus isolation media, not only was SK2 agar capable of selectively recovering guaiacol-producing Alicyclobacillus, the degree of growth was also approximately equal if not better than orange serum agar, potato dextrose agar, and K agar. The development of SK2 agar provides the fruit juice industry with an inexpensive, simple to use alternative for the detection of guaiacol producing Alicyclobacillus.

  8. Characteristics of thermoplastic sugar palm Starch/Agar blend: Thermal, tensile, and physical properties.

    PubMed

    Jumaidin, R; Sapuan, S M; Jawaid, M; Ishak, M R; Sahari, J

    2016-08-01

    The aim of this work is to study the behavior of biodegradable sugar palm starch (SPS) based thermoplastic containing agar in the range of 10-40wt%. The thermoplastics were melt-mixed and then hot pressed at 140°C for 10min. SEM investigation showed good miscibility between SPS and agar. FT-IR analysis confirmed that SPS and agar were compatible and inter-molecular hydrogen bonds existed between them. Incorporation of agar increased the thermoplastic starch tensile properties (Young's modulus and tensile strength). The thermal stability and moisture uptake increased with increasing agar content. The present work shows that starch-based thermoplastics with 30wt% agar content have the highest tensile strength. Higher content of agar (40wt%) resulted to more rough cleavage fracture and slight decrease in the tensile strength. In conclusion, the addition of agar improved the thermal and tensile properties of thermoplastic SPS which widened the potential application of this eco-friendly material. The most promising applications for this eco-friendly material are short-life products such as packaging, container, tray, etc.

  9. Inhibition of Streptococcus mutans strains by different mitis-salivarius agar preparations.

    PubMed Central

    Staat, R H

    1976-01-01

    Several Streptococcus mutans strains were markedly inhibited by mitis-salivarius agar manufactured by Baltimore Biological Laboratories, but little, if any, inhibition was noted using Difco Laboratories' mitis-salivarius agar. Supplementation of the basic medium with sucrose and bacitracin for specific selection of S. mutans resulted in suppression of representative S. mutans type a strains regardless of manufacturer. PMID:1270597

  10. Characteristics of thermoplastic sugar palm Starch/Agar blend: Thermal, tensile, and physical properties.

    PubMed

    Jumaidin, R; Sapuan, S M; Jawaid, M; Ishak, M R; Sahari, J

    2016-08-01

    The aim of this work is to study the behavior of biodegradable sugar palm starch (SPS) based thermoplastic containing agar in the range of 10-40wt%. The thermoplastics were melt-mixed and then hot pressed at 140°C for 10min. SEM investigation showed good miscibility between SPS and agar. FT-IR analysis confirmed that SPS and agar were compatible and inter-molecular hydrogen bonds existed between them. Incorporation of agar increased the thermoplastic starch tensile properties (Young's modulus and tensile strength). The thermal stability and moisture uptake increased with increasing agar content. The present work shows that starch-based thermoplastics with 30wt% agar content have the highest tensile strength. Higher content of agar (40wt%) resulted to more rough cleavage fracture and slight decrease in the tensile strength. In conclusion, the addition of agar improved the thermal and tensile properties of thermoplastic SPS which widened the potential application of this eco-friendly material. The most promising applications for this eco-friendly material are short-life products such as packaging, container, tray, etc. PMID:27177458

  11. Electrospinning of agar/PVA aqueous solutions and its relation with rheological properties.

    PubMed

    Sousa, Ana M M; Souza, Hiléia K S; Uknalis, Joseph; Liu, Shih-Chuan; Gonçalves, Maria P; Liu, LinShu

    2015-01-22

    In this work, we report the successful fabrication of agar-based nanofibers by electrospinning technique, using water as solvent media. A tubeless spinneret was attached inside the electrospinning chamber, operating at 50°C, to avoid agar gelation. Agar pure solution (1 wt%) showed inadequate spinnability regardless of the used electrospinning conditions. The addition of a co-blending polymer such as PVA (10 wt% starting solution) improved the solutions viscoelasticity and hence, the solutions spinnability. Agar/PVA solutions were prepared with different mass ratios (100/0, 50/50, 40/60, 30/70, 20/80 and 0/100) and electrospun at various sets of electrospinning conditions. Best nanofibers were obtained with 30/70 and 20/80 agar/PVA blends while samples with higher agar contents (50/50 and 40/60 agar/PVA) were harder to process and led to discontinuous fibrous mats. This first set of encouraging results can open a new window of opportunities for agar-based biomaterials in the form of nanofibers.

  12. Efficacy of agar-plate culture in detection of Strongyloides stercoralis infection.

    PubMed

    Arakaki, T; Iwanaga, M; Kinjo, F; Saito, A; Asato, R; Ikeshiro, T

    1990-06-01

    Agar-plate culture of feces using a modified petri dish proved to be highly efficient in the detection of Strongyloides stercoralis infection. Furrows left by S. stercoralis on the agar plate were distinguished readily in size from those left by Necator americanus. PMID:2352073

  13. Dilution method study on the interfacial composition, thermodynamic properties and structural parameters of W/O microemulsions stabilized by 1-pentanol and surfactants in absence and presence of sodium chloride.

    PubMed

    Paul, Bidyut K; Nandy, Debdurlav

    2007-12-15

    The phase behaviors, interfacial composition, thermodynamic properties and structural characteristics of water-in-oil microemulsions under varied molar ratio of water to surfactant (omega) at 303 K and also by varying temperatures at a fixed omega(=40) by mixing with 1-pentanol and decane or dodecane in absence and presence of sodium chloride have been studied by the method of dilution. The surfactants used were cetyl pyridinium chloride (CPC), sodium dodecyl sulfate (SDS) and polyoxyethylene (23) lauryl ether (Brij-35). The compositions of 1-pentanol and the surfactant at the interfacial region, the distribution of 1-pentanol between the interfacial region and the continuous oil phase, and the effective packing parameter (P(eff)) at the threshold level of stability have been estimated. The thermodynamics of transfer of 1-pentanol from the continuous oil phase to the interface have been evaluated. The structural parameters viz. radii of the droplet (R(e)) and the waterpool (R(w)), effective thickness of the interfacial layer (d(I)), average aggregation numbers of surfactants (N (s)) and the cosurfactant (1-pentanol) (N (a)) and the number of droplets (N(d)) have also been estimated. The prospect of using these w/o microemulsions for the synthesis of nanoparticles with small size, have been discussed in the light of the radii of the droplet, and waterpool, the extent of variation of effective thickness of the droplet under varied molar ratio of water to surfactant and temperature. An attempt has been made to rationalize the results in a comprehensive manner.

  14. [Evaluation of a new medium, eggplant (Solanum melongena) agar as a screening medium for Cryptococcus neoformans in environmental samples].

    PubMed

    Sengul, Mustafa; Ergin, Cağrı; Kartal, Tuğba

    2014-04-01

    Cryptococcus neofomans is an encapsulated yeast-like fungus that causes life-threatening infections, especially in immunosuppresive patients. C.neoformans infection is believed to be acquired via inhalation of aerosolized particles from the environment. Avian guano, decaying tree hollows and soil are the related known environmental niches. Brown pigmented yeast growth from the precursors in growth media is an important step for the identification and isolation of C.neoformans. Seeds of plants in nature are preferred owing to easy accessibility and low costs for the preparation of such media. Guizotia abysinicca (Niger seed) as Staib agar, Helianthus annus (Sunflower) as Pal's medium, Brassica nigra (Mustard) agar, tobacco agar, Mucuna pruriens (Velvet bean) seed agar, Perilla frutescens (Beefsteak plant) seed agar, Rubus fruticosus (Blackberry) agar and ground red hot pepper agar are pigment-based selective media for the differentiation of C.neoformans. The aim of this study was to observe the pigment production of C.neoformans in a new medium based on eggplant (Solanum melongena) and also to compare its performance with the simplified Staib, Pal's and tobacco agar for isolation from the environment. Three different eggplant-based medium (S.melongena Melanzaza viserba, S.melongena Pinstripe F1 and S.ovigerum Ivory F1) were included in the study. Pigment-forming eggplant medium, simplified Staib agar, Pal's agar and tobacco agar were used for the cultivation of the environmental swabbed samples from 19 Eucalyptus camaldulensis trunk hollows in continuous colonization region. While pigment formation were observed with S.melongena Melanzaza viserba and S.melongena Pinstripe F1 containing media, S.ovigerum Ivory F1 medium was found to be non-reactive. In colonization area (Gökova-Akyaka, Turkey), 11 (57.9%) out of 19 E.camaldulensis samples were positive with simplified Staib agar, Pal's agar and eggplant agar while 10 (52.6%) of them are positive with tobacco agar. C

  15. [Evaluation of a new medium, eggplant (Solanum melongena) agar as a screening medium for Cryptococcus neoformans in environmental samples].

    PubMed

    Sengul, Mustafa; Ergin, Cağrı; Kartal, Tuğba

    2014-04-01

    Cryptococcus neofomans is an encapsulated yeast-like fungus that causes life-threatening infections, especially in immunosuppresive patients. C.neoformans infection is believed to be acquired via inhalation of aerosolized particles from the environment. Avian guano, decaying tree hollows and soil are the related known environmental niches. Brown pigmented yeast growth from the precursors in growth media is an important step for the identification and isolation of C.neoformans. Seeds of plants in nature are preferred owing to easy accessibility and low costs for the preparation of such media. Guizotia abysinicca (Niger seed) as Staib agar, Helianthus annus (Sunflower) as Pal's medium, Brassica nigra (Mustard) agar, tobacco agar, Mucuna pruriens (Velvet bean) seed agar, Perilla frutescens (Beefsteak plant) seed agar, Rubus fruticosus (Blackberry) agar and ground red hot pepper agar are pigment-based selective media for the differentiation of C.neoformans. The aim of this study was to observe the pigment production of C.neoformans in a new medium based on eggplant (Solanum melongena) and also to compare its performance with the simplified Staib, Pal's and tobacco agar for isolation from the environment. Three different eggplant-based medium (S.melongena Melanzaza viserba, S.melongena Pinstripe F1 and S.ovigerum Ivory F1) were included in the study. Pigment-forming eggplant medium, simplified Staib agar, Pal's agar and tobacco agar were used for the cultivation of the environmental swabbed samples from 19 Eucalyptus camaldulensis trunk hollows in continuous colonization region. While pigment formation were observed with S.melongena Melanzaza viserba and S.melongena Pinstripe F1 containing media, S.ovigerum Ivory F1 medium was found to be non-reactive. In colonization area (Gökova-Akyaka, Turkey), 11 (57.9%) out of 19 E.camaldulensis samples were positive with simplified Staib agar, Pal's agar and eggplant agar while 10 (52.6%) of them are positive with tobacco agar. C

  16. Mechanical touch responses of Arabidopsis TCH1-3 mutant roots on inclined hard-agar surface

    NASA Astrophysics Data System (ADS)

    Zha, Guodong; Wang, Bochu; Liu, Junyu; Yan, Jie; Zhu, Liqing; Yang, Xingyan

    2016-01-01

    The gravity-induced mechanical touch stimulus can affect plant root architecture. Mechanical touch responses of plant roots are an important aspect of plant root growth and development. Previous studies have reported that Arabidopsis TCH1-3 genes are involved in mechano-related events, how-ever, the physiological functions of TCH1-3 genes in Arabidopsis root mechanoresponses remain unclear. In the present study, we applied an inclined hard agar plate method to produce mechanical touch stimulus, and provided evidence that altered mechanical environment could influence root growth. Furthermore, tch1-3 Arabidopsis mutants were investigated on inclined agar surfaces to explore the functions of TCH1-3 genes on Arabidopsis root mechanoresponses. The results showed that two tch2 mutants, cml24-2 and cml24-4, exhibited significantly reduced root length, biased skewing, and decreased density of lateral root. In addition, primary root length and density of lateral root of tch3 (cml12-2) was significantly decreased on inclined agar surfaces. This study indicates that the tch2 and tch3 mutants are hypersensitive to mechanical touch stimulus, and TCH2 (CML24-2 and CML24-4) and TCH3 (CML12-2) genes may participate in the mechanical touch response of Arabidopsis roots.

  17. Dilute Acid and Autohydrolysis Pretreatment

    NASA Astrophysics Data System (ADS)

    Yang, Bin; Wyman, Charles E.

    Exposure of cellulosic biomass to temperatures of about 120-210°C can remove most of the hemicellulose and produce cellulose-rich solids from which high glucose yields are possible with cellulase enzymes. Furthermore, the use of dilute sulfuric acid in this pretreatment operation can increase recovery of hemicellulose sugars substantially to about 85-95% of the maximum possible versus only about 65% if no acid is employed. The use of small-diameter tubes makes it possible to employ high solids concentrations similar to those preferred for commercial operations, with rapid heat-up, good temperature control, and accurate closure of material balances. Mixed reactors can be employed to pretreat larger amounts of biomass than possible in such small-diameter tubes, but solids concentrations are limited to about 15% or less to provide uniform temperatures. Pretreatment of large amounts of biomass at high solids concentrations is best carried out using direct steam injection and rapid pressure release, but closure of material balances in such “steam gun” devices is more difficult. Although flow of water alone or containing dilute acid is not practical commercially, such flow-through configurations provide valuable insight into biomass deconstruction kinetics not possible in the batch tubes, mixed reactors, or steam gun systems.

  18. A Microgravity Helium Dilution Cooler

    NASA Technical Reports Server (NTRS)

    Roach, Pat R.; Sperans, Joel (Technical Monitor)

    1994-01-01

    We are developing a He-3-He-4 dilution cooler to operate in microgravity. It uses charcoal adsorption pumps and heaters for its operation; it has no moving parts. It currently operates cyclically to well below 0.1 K and we have designed a version to operate continuously. We expect that the continuous version will be able to provide the long-duration cooling that many experiments need at temperatures down to 0.040 K. More importantly, such a dilution cooler could provide the precooling that enables the use of adiabatic demagnetization techniques that can reach temperatures below 0.001 K. At temperatures below 0.002 K many fascinating microgravity experiments on superfluid He-3 become possible. Among the possibilities are: research into a superfluid He-3 gyroscope, study of the nucleation of the B-phase of superfluid He-3 when the sample is floating out of contact with walls, study of the anisotropy of the surface tension of the B-phase, and NMR experiments on tiny free-floating clusters of superfluid He-3 atoms that should model the shell structure of nuclei.

  19. A preliminary evaluation of a new selective agar supplemented with desferrioxamine for detection of methicillin-resistant Staphylococcus aureus.

    PubMed

    Monsen, T; Olofsson, C; Granström, S; Wiström, J

    2003-07-01

    The aim of the present study was to evaluate the performance of two new selective screening agars, Colombia agar supplemented with 1000 mg/L desferrioxamine, 5 mg/L amphotericin B, 16 mg/L polymyxin B, and 2 mg/L methicillin (CMDAP agar) or 0.5 mg/L oxacillin (CODAP agar), for detection of methicillin-resistant Staphylococcus aureus (MRSA). Both the CMDAP and the CODAP agar effectively inhibited growth of 151 isolates of coagulase-negative staphylococci (CoNS), 45 of Enterobacteriaceae and six Candida spp. examined. The sensitivity and specificity of the CMDAP and CODAP agars for detection of MRSA was calculated by comparing the growth of 52 MRSA with the inhibition of 74 mecA negative S. aureus and of 151 CoNS. The performance of the new agars was compared with four previously described MRSA screening agars. The sensitivity and specificity for detection of MRSA after incubation at 35 degrees C for 24 h was 0.94 and 0.91, respectively, for the CMDAP agar, 0.60 and 0.90 for the CODAP agar, 0.98 and 0.57 for methicillin aztreonam mannitol salt agar (MAMSA), 0.23 and 0.84 for oxacillin mannitol salt agar (OMSA), 0.48 and 0.76 for oxacillin Mueller-Hinton agar (OMHA) and 0.75 and 0.77 for lithium oxacillin mannitol salt agar (LOMSA). Agars supplemented with desferrioxamine, CMDAP and CODAP, were more specific for detecting MRSA compared with agars not supplemented with desferrioxamine. The detection rate was higher for agars supplemented with methicillin than for agars supplemented with oxacillin.

  20. Maximum independent set on diluted triangular lattices

    NASA Astrophysics Data System (ADS)

    Fay, C. W., IV; Liu, J. W.; Duxbury, P. M.

    2006-05-01

    Core percolation and maximum independent set on random graphs have recently been characterized using the methods of statistical physics. Here we present a statistical physics study of these problems on bond diluted triangular lattices. Core percolation critical behavior is found to be consistent with the standard percolation values, though there are strong finite size effects. A transfer matrix method is developed and applied to find accurate values of the density and degeneracy of the maximum independent set on lattices of limited width but large length. An extrapolation of these results to the infinite lattice limit yields high precision results, which are tabulated. These results are compared to results found using both vertex based and edge based local probability recursion algorithms, which have proven useful in the analysis of hard computational problems, such as the satisfiability problem.

  1. Multicenter comparison of the ISO standard 20776-1 and the serial 2-fold dilution procedures to dilute hydrophilic and hydrophobic antifungal agents for susceptibility testing.

    PubMed

    Gomez-Lopez, Alicia; Arendrup, Maiken Cavling; Lass-Floerl, Cornelia; Rodriguez-Tudela, Juan-Luis; Cuenca-Estrella, Manuel

    2010-05-01

    A multicenter study was conducted to assess the accuracy of the ISO standard 20776-1 and the serial 2-fold dilution procedures for antifungal susceptibility testing. Fluconazole trays can be accurately prepared by following ISO and serial dilution schemes. However, itraconazole trays showed a significant lack of reproducibility that was independent of which method was followed.

  2. Evaluation of use of a new chromogenic agar in detection of urinary tract pathogens.

    PubMed

    Samra, Z; Heifetz, M; Talmor, J; Bain, E; Bahar, J

    1998-04-01

    CHROMagar Orientation, a new chromogenic medium, was evaluated for the detection and differentiation of gram-positive and gram-negative pathogenic microorganisms in 900 urine samples from hospitalized patients. Performance characteristics of the medium were evaluated in comparison to those of 5% sheep blood and MacConkey agars by direct inoculation of the urine samples on the three media. Four gram-negative and two gram-positive strains as well as one yeast control strain from the American Type Culture Collection were used to ensure quality control. CHROMagar Orientation succeeded in detecting all the urine pathogens that were detected by the reference media, including gram-negative bacilli, staphylococci, streptococci, and yeasts. Colony color and morphology on CHROMagar Orientation accurately differentiated Escherichia coli, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, and Acinetobacter spp. Owing to the similarity in the pigmentation produced by Klebsiella, Enterobacter, and Citrobacter isolates, the medium failed to distinguish among them; however, these isolates were easily recognized as coliforms because of their metallic blue coloration. Staphylococci were clearly perceptible: S. aureus and S. epidermidis grow in regular-size colonies that range from opaque white to yellowish, and S. saprophyticus produces opaque pink colonies. All streptococcus strains, including those from groups B and C, were detected. They grow as undifferentiated flat dry diffused colonies, and additional tests were required for identification. Enterococci were easily discriminated by their strong turquoise pigmentation and their typical growth on the agar's surface. Yeast grow in typical creamy wet convex colonies. The accuracy of antibiotic susceptibility determinations according to standard methods was also tested by picking isolates directly from CHROMagar Orientation. The results showed excellent correlation with those obtained with microorganisms picked from

  3. XAFS in dilute magnetic semiconductors.

    PubMed

    Sun, Zhihu; Yan, Wensheng; Yao, Tao; Liu, Qinghua; Xie, Yi; Wei, Shiqiang

    2013-10-14

    X-Ray absorption fine structure (XAFS) spectroscopy has experienced a rapid development in the last four decades and has proved to be a powerful structure characterization technique in the study of local environments in condensed matter. In this article, we first introduce the XAFS basic principles including theory, data analysis and experiment in some detail. Then we attempt to make a review on the applications of XAFS to the study of atomic and electronic structure in dilute magnetic semiconductor (DMS) systems. The power of XAFS in characterizing this interesting material system, such as determining the occupation sites and distribution of the dopants, detecting the presence of metal clusters or secondary phases, as well as identifying the defect types and dopant valence, will be illuminated by selected examples. This review should be of interest both to newcomers in the DMS field and to an interdisciplinary community of researchers working in synthesis, characterization and utilization of DMS materials. PMID:23884341

  4. Desynchronization in diluted neural networks

    SciTech Connect

    Zillmer, Ruediger; Livi, Roberto; Politi, Antonio; Torcini, Alessandro

    2006-09-15

    The dynamical behavior of a weakly diluted fully inhibitory network of pulse-coupled spiking neurons is investigated. Upon increasing the coupling strength, a transition from regular to stochasticlike regime is observed. In the weak-coupling phase, a periodic dynamics is rapidly approached, with all neurons firing with the same rate and mutually phase locked. The strong-coupling phase is characterized by an irregular pattern, even though the maximum Lyapunov exponent is negative. The paradox is solved by drawing an analogy with the phenomenon of 'stable chaos', i.e., by observing that the stochasticlike behavior is 'limited' to an exponentially long (with the system size) transient. Remarkably, the transient dynamics turns out to be stationary.

  5. Band anticrossing in dilute nitrides

    SciTech Connect

    Shan, W.; Yu, K.M.; Walukiewicz, W.; Wu, J.; Ager III, J.W.; Haller, E.E.

    2003-12-23

    Alloying III-V compounds with small amounts of nitrogen leads to dramatic reduction of the fundamental band-gap energy in the resulting dilute nitride alloys. The effect originates from an anti-crossing interaction between the extended conduction-band states and localized N states. The interaction splits the conduction band into two nonparabolic subbands. The downward shift of the lower conduction subband edge is responsible for the N-induced reduction of the fundamental band-gap energy. The changes in the conduction band structure result in significant increase in electron effective mass and decrease in the electron mobility, and lead to a large enhance of the maximum doping level in GaInNAs doped with group VI donors. In addition, a striking asymmetry in the electrical activation of group IV and group VI donors can be attributed to mutual passivation process through formation of the nearest neighbor group-IV donor nitrogen pairs.

  6. XAFS in dilute magnetic semiconductors.

    PubMed

    Sun, Zhihu; Yan, Wensheng; Yao, Tao; Liu, Qinghua; Xie, Yi; Wei, Shiqiang

    2013-10-14

    X-Ray absorption fine structure (XAFS) spectroscopy has experienced a rapid development in the last four decades and has proved to be a powerful structure characterization technique in the study of local environments in condensed matter. In this article, we first introduce the XAFS basic principles including theory, data analysis and experiment in some detail. Then we attempt to make a review on the applications of XAFS to the study of atomic and electronic structure in dilute magnetic semiconductor (DMS) systems. The power of XAFS in characterizing this interesting material system, such as determining the occupation sites and distribution of the dopants, detecting the presence of metal clusters or secondary phases, as well as identifying the defect types and dopant valence, will be illuminated by selected examples. This review should be of interest both to newcomers in the DMS field and to an interdisciplinary community of researchers working in synthesis, characterization and utilization of DMS materials.

  7. Effectual detection of group B streptococci with reduced penicillin susceptibility (PRGBS) by commercially available methicillin-resistant-Staphylococcus aureus (MRSA)-selective agar.

    PubMed

    Fukigai, Shinako; Morimoto, Makiko; Kimura, Kouji; Doyama, Yo; Miyazaki, Akira; Kamiya, Chitose; Banno, Hirotsugu; Morishima, Eriko; Onoda, Tomohiro; Nagano, Noriyuki; Jin, Wanchun; Wachino, Jun-Ichi; Yamada, Keiko; Arakawa, Yoshichika

    2016-07-01

    We evaluated the feasibility and efficacy of a commercially available methicillin-resistant Staphylococcus aureus (MRSA)-selective agar, chromID(™) MRSA, to detect group B streptococci with reduced penicillin susceptibility (PRGBS) in this study. The results showed 72.4% (21/29) sensitivity and 98.4% (60/61) specificity to detect PRGBS using this method.

  8. Total body water and lean body mass estimated by ethanol dilution

    NASA Technical Reports Server (NTRS)

    Loeppky, J. A.; Myhre, L. G.; Venters, M. D.; Luft, U. C.

    1977-01-01

    A method for estimating total body water (TBW) using breath analyses of blood ethanol content is described. Regression analysis of ethanol concentration curves permits determination of a theoretical concentration that would have existed if complete equilibration had taken place immediately upon ingestion of the ethanol; the water fraction of normal blood may then be used to calculate TBW. The ethanol dilution method is applied to 35 subjects, and comparison with a tritium dilution method of determining TBW indicates that the correlation between the two procedures is highly significant. Lean body mass and fat fraction were determined by hydrostatic weighing, and these data also prove compatible with results obtained from the ethanol dilution method. In contrast to the radioactive tritium dilution method, the ethanol dilution method can be repeated daily with its applicability ranging from diseased individuals to individuals subjected to thermal stress, strenuous exercise, water immersion, or the weightless conditions of space flights.

  9. Dilution in single pass arc welds

    SciTech Connect

    DuPont, J.N.; Marder, A.R.

    1996-06-01

    A study was conducted on dilution of single pass arc welds of type 308 stainless steel filler metal deposited onto A36 carbon steel by the plasma arc welding (PAW), gas tungsten arc welding (GTAW), gas metal arc welding (GMAW), and submerged arc welding (SAW) processes. Knowledge of the arc and melting efficiency was used in a simple energy balance to develop an expression for dilution as a function of welding variables and thermophysical properties of the filler metal and substrate. Comparison of calculated and experimentally determined dilution values shows the approach provides reasonable predictions of dilution when the melting efficiency can be accurately predicted. The conditions under which such accuracy is obtained are discussed. A diagram is developed from the dilution equation which readily reveals the effect of processing parameters on dilution to aid in parameter optimization.

  10. Development of an improved selective agar medium for isolation of Yersinia pestis.

    PubMed

    Ber, Raphael; Mamroud, Emanuelle; Aftalion, Moshe; Tidhar, Avital; Gur, David; Flashner, Yehuda; Cohen, Sara

    2003-10-01

    Existing media designed for selective isolation of clinically important members of the genus Yersinia were found to be unsatisfactory for the growth and isolation of Yersinia pestis. We report the development of a new selective agar medium (termed BIN) that supports the growth of Y. pestis. The development of the formulation of this medium was based on a fluorescence screening system designed for monitoring bacterial growth on semisolid media, using a green fluorescent protein-expressing strain. High-throughput combinatorial experiments can be conducted for the quantitative evaluation of the effect of different medium components on growth. Generation of fluorescence plots in this system, using microplates, allowed the quantitative evaluation of the growth rate of Y. pestis EV76 cultures in different agar compositions. The final BIN formulation is based on brain heart infusion agar, to which the selective agents irgasan, cholate salts, crystal violet, and nystatin were introduced. It was found that BIN agar is more efficient in supporting colony formation and recovery of Y. pestis than are the conventional semisolid media MacConkey agar and Yersinia-selective agar (cefsulodin-irgasan-novobiocin agar). The advantage of BIN over other media has been also demonstrated in recovering virulent Y. pestis from the mixed bacterial populations found in decaying carcasses of infected mice. The BIN medium is suggested as a selective medium for isolation and recovery of Y. pestis from various backgrounds.

  11. Is a New Standard Needed for Diffusion Methods for In Vitro Susceptibility Testing of Fosfomycin against Pseudomonas aeruginosa?

    PubMed Central

    Díez-Aguilar, María; Martínez-García, Laura; Morosini, María Isabel

    2015-01-01

    We analyzed fosfomycin susceptibility results in Pseudomonas aeruginosa clinical isolates obtained by MIC gradient strips and disk diffusion methods using two different inocula, 108 and 106 CFU/ml, and compared them to the agar dilution reference method. Essential and categorical agreements were 93.6% and 95%, respectively, for the 106 CFU/ml alternative inoculum, and they were 67.6% and 78.2%, respectively, for the standard inoculum (108 CFU/ml). The use of the 106 CFU/ml inoculum improves the agreement values and inhibition zone readings. PMID:26643341

  12. Is a New Standard Needed for Diffusion Methods for In Vitro Susceptibility Testing of Fosfomycin against Pseudomonas aeruginosa?

    PubMed

    Díez-Aguilar, María; Martínez-García, Laura; Cantón, Rafael; Morosini, María Isabel

    2016-02-01

    We analyzed fosfomycin susceptibility results in Pseudomonas aeruginosa clinical isolates obtained by MIC gradient strips and disk diffusion methods using two different inocula, 10(8) and 10(6) CFU/ml, and compared them to the agar dilution reference method. Essential and categorical agreements were 93.6% and 95%, respectively, for the 10(6) CFU/ml alternative inoculum, and they were 67.6% and 78.2%, respectively, for the standard inoculum (10(8) CFU/ml). The use of the 10(6) CFU/ml inoculum improves the agreement values and inhibition zone readings.

  13. Evaluation of heavy water for indicator dilution cardiac output measurement

    SciTech Connect

    Schreiner, M.S.; Leksell, L.G.; Neufeld, G.R. )

    1989-10-01

    We evaluated deuterium oxide (D2O) as a tracer for cardiac output measurements. Cardiac output measurements made by thermodilution were compared with those made by indicator dilution with D2O and indocyanine green as tracers. Five triplicate measurements for each method were made at intervals of 30 minutes in each of 9 anesthetized, mechanically ventilated goats. Cardiac output ranged between 0.68 and 3.79 L/min. The 45 data points yielded a correlation coefficient of 0.948 for the comparison of D2O indicator dilution cardiac output measurements with thermodilution measurements and a linear regression slope of 1.046. D2O indicator dilution measurements were biased by -0.11 +/- 0.22 L/min compared with thermodilution measurements and had a standard deviation of +/- 0.12 L/min for triplicate measurements. Hematocrits ranging between 20 and 50 vol% had no effect on optical density for D2O. D2O is more stable than indocyanine green and approximately one-tenth the price (40 cents per injection compared with $4). The basic instrumentation cost of approximately $9,000 is an additional initial expense, but provides the ability to perform pulmonary extravascular water measurements with a double-indicator dilution technique. D2O has potential as a tracer for the clinical determination of indicator dilution cardiac output measurements and pulmonary extravascular water measurements.

  14. [Formation of oxalate in oxaliplatin injection diluted with infusion solutions].

    PubMed

    Eto, Seiji; Yamamoto, Kie; Shimazu, Kounosuke; Sugiura, Toshimune; Baba, Kaori; Sato, Ayaka; Goromaru, Takeshi; Hagiwara, Yoshiaki; Hara, Keiko; Shinohara, Yoshitake; Takahashi, Kojiro

    2014-01-01

    Oxaliplatin use can cause acute peripheral neuropathy characterized by sensory paresthesias, which are markedly exacerbated by exposure to cold temperatures, and is a dose-limiting factor in the treatment of colorectal cancer.Oxalate is eliminated in a series of nonenzymatic conversions of oxaliplatin in infusion solutions or biological fluids.Elimination of oxalate from oxaliplatin has been suggested as one of the reasons for the development of acute neuropathy.In this study, we developed a high-performance liquid chromatography(HPLC)-based method to detect oxalate formation, and investigated the time dependent formation of oxalate in oxaliplatin diluted with infusion solutions.The results obtained showed that the amount of oxalate in the solution corresponded to 1.6% of oxaliplatin 8 h after oxaliplatin dilution with a 5% glucose solution. On the other hand, oxalate formation from oxaliplatin diluted with a saline solution was ten-fold higher than that from oxaliplatin diluted with the 5% glucose solution.Most patients who were intravenously injected with oxaliplatin experienced venous pain.As a preventive measure against venous pain, dexamethasone was added to the oxaliplatin injection.We measured the amount of oxalate formed in the dexamethasone-containing oxaliplatin injection diluted with a 5% glucose solution.The amount of oxalate formed when dexamethasone was added did not differ significantly from that formed when dexamethasone was not added.Thus, there are no clinical problems associated with the stability of oxaliplatin solutions.

  15. Xanthan gum: an economical substitute for agar in plant tissue culture media.

    PubMed

    Jain, R; Babbar, S B

    2006-03-01

    Xanthan gum, a microbial desiccation-resistant polysaccharide prepared commercially by aerobic submerged fermentation from Xanthomonas campestris, has been successfully used as a solidifying agent for plant tissue culture media. Its suitability as a substitute to agar was demonstrated for in vitro seed germination, caulogenesis and rhizogenesis of Albizzia lebbeck, androgenesis in anther cultures of Datura innoxia, and somatic embryogenesis in callus cultures of Calliandra tweedii. Culture media used for eliciting these morphogenic responses were gelled with either 1% xanthan gum or 0.9% agar. Xanthan gum, like agar, supported all these responses.

  16. Polymer film deposition on agar using a dielectric barrier discharge jet and its bacterial growth inhibition

    NASA Astrophysics Data System (ADS)

    Tsai, T.-C.; Cho, J.; Mcintyre, K.; Jo, Y.-K.; Staack, D.

    2012-08-01

    Polymer film deposition on agar in ambient air was achieved using the helium dielectric barrier discharge jet (DBD jet) fed with polymer precursors, and the bacterial growth inhibition due to the deposited film was observed. The DBD jet with precursor addition was more efficient at sterilization than a helium-only DBD jet. On the areas where polymer films cover the agar the bacterial growth was significantly inhibited. The inhibition efficacy showed dependence on the film thickness. The DBD jet without precursor also created a modified agar layer, which may slow the growth of some bacterial strains.

  17. A note on a selective agar medium for the enumeration of Flavobacterium species in water.

    PubMed

    Flint, K P

    1985-12-01

    A selective nutrient agar medium containing kanamycin at 50 micrograms/ml was developed for the isolation and enumeration of yellow-pigmented colonies from the River Sowe, Coventry. Such organisms were shown to be members of the heterogeneous genus Flavobacterium. Typically, yellow pigmented colonies constituted less than 10% of the colonies on nutrient agar alone but up to 70% on nutrient agar plus kanamycin. This medium is a useful addition to the range of media available for the isolation and further ecological study of particular species of this important group of micro-organisms.

  18. Diluted magnetic semiconductors with narrow band gaps

    NASA Astrophysics Data System (ADS)

    Gu, Bo; Maekawa, Sadamichi

    2016-10-01

    We propose a method to realize diluted magnetic semiconductors (DMSs) with p - and n -type carriers by choosing host semiconductors with a narrow band gap. By employing a combination of the density function theory and quantum Monte Carlo simulation, we demonstrate such semiconductors using Mn-doped BaZn2As2 , which has a band gap of 0.2 eV. In addition, we found a nontoxic DMS Mn-doped BaZn2Sb2 , of which the Curie temperature Tc is predicted to be higher than that of Mn-doped BaZn2As2 , the Tc of which was up to 230 K in a recent experiment.

  19. [Comparison of the results of determining antibiotic sensitivity on AGV medium and on Mueller-Hinton and isosensitest agars].

    PubMed

    Kozlov, R S; Strachunskiĭ, L S; Livermor, D M; Stetsiuk, O U; Shavrikova, E P

    1996-01-01

    To evaluate the adequacy of AGV agar for antimicrobial susceptibility testing, the susceptibility of a range of bacteria to 10 antimicrobials on AGV, Mueller-Hinton and isoSensitest agars, all supplemented with 5 per cent lyzed horse blood was determined. Disc tests were used. In general, AGV agar gave identical susceptibility results to Mueller-Hinton and isoSensitest agars for common gram positive and gram negative bacteria with most of the tested microbials excluding sulphonamides and trimethoprim. With those latter antimicrobials inhibition zones for susceptible organisms were not formed on AGV agar whereas large zones were present on Mueller-Hinton and isoSensitest agars. This discrepancy probably can be explained by the presence of high levels of thymidine in AGV agar; too high to be corrected even by the addition of 5 per cent lysed horse blood. AGV agar is possible to use for susceptibility testing with most of the microbials excluding trimethoprim and sulphonamides.

  20. Gas dilution system results and application to acid rain utilities

    SciTech Connect

    Jolley-Souders, K.; Geib, R.; Dunn, C.

    1997-12-31

    In 1997, the United States EPA will remove restrictions preventing acid rain utilities from using gas dilution systems for calibration or linearity studies for continuous emissions monitoring, Test Method 205 in 40CFR51 requires that a gas dilution system must produce calibration gases whose measured values are within {+-}2% of predicted values. This paper presents the evaluation of the Environics/CalMat 2020 Dilution System for use in calibration studies. Internal studies show that concentrations generated by this unit are within {+-}0.5% of predicted values. Studies are being conducted by several acid rain utilities to evaluate the Environics/CalMat system using single minor component calibration standards. In addition, an internally generated study is being performed to demonstrate the system`s accuracy using a multi-component gas mixture. Data from these tests will be presented in the final version of the paper.

  1. Growth characteristics of Bacillus anthracis compared to other Bacillus spp. on the selective nutrient media Anthrax Blood Agar and Cereus Ident Agar.

    PubMed

    Tomaso, Herbert; Bartling, Carsten; Al Dahouk, Sascha; Hagen, Ralf M; Scholz, Holger C; Beyer, Wolfgang; Neubauer, Heinrich

    2006-01-01

    Anthrax Blood Agar (ABA) and Cereus Ident Agar (CEI) were evaluated as selective growth media for the isolation of Bacillus anthracis using 92 B. anthracis and 132 other Bacillus strains from 30 species. The positive predictive values for the identification of B. anthracis on ABA, CEI, and the combination of both were 72%, 71%, and 90%, respectively. Thus, less than 10% of all species were misidentified using both nutrient media. Species which might be misidentified as B. anthracis were B. cereus, B. mycoides, and B. thuringiensis. Particularly, 30% of B. weihenstephanensis strains were misidentified as B. anthracis.

  2. Reusable chelating resins concentrate metal ions from highly dilute solutions

    NASA Technical Reports Server (NTRS)

    Bauman, A. J.; Weetal, H. H.; Weliky, N.

    1966-01-01

    Column chromatographic method uses new metal chelating resins for recovering heavy-metal ions from highly dilute solutions. The absorbed heavy-metal cations may be removed from the chelating resins by acid or base washes. The resins are reusable after the washes are completed.

  3. Neonothopanus gardneri: a new combination for a bioluminescent agaric from Brazil.

    PubMed

    Capelari, Marina; Desjardin, Dennis E; Perry, Brian A; Asai, Tatiane; Stevani, Cassius V

    2011-01-01

    The bioluminescent agaric, Agaricus gardneri Berk., was rediscovered recently in central Brazil. The new combination, Neonothopanus gardneri, is proposed for this long-forgotten taxon supported by morphological and molecular data.

  4. A fresh liver agar substrate for rearing small numbers of forensically important blow flies (Diptera: Calliphoridae)

    USGS Publications Warehouse

    Gruner, Susan V.; Slone, Daniel H.

    2014-01-01

    Forensically important calliphorids can be reared on a mixture of beef liver and agar. Small pieces of meat, especially fresh or frozen beef liver, will desiccate in 2–6 h, but this simple-to-make feeding substrate remains moist for at least 12 h at 25 and 30°C without desiccation, even in small (5 g) amounts. We determined the survivorship of small numbers of Chrysomya megacephala (F.) (first-instar larvae to adult eclosion) raised on 5 g of liver agar and fresh beef liver. We found that all larvae raised on 5 g of liver died due to desiccation, but survivorship on 5 g of liver agar was equivalent to that on larger (50 g) pieces of either liver agar or beef liver.

  5. [THE APPLICATION OF SELECTIVE CHROMOGENIC AGAR FOR DETECTING ENTEROBACTERIA WITH PRODUCTION OF BETA-LACTAMASES].

    PubMed

    Korobova, A G; Frolova, L N; Kliasova, G A

    2015-11-01

    The detection of enterobacteria with production of beta-lactamases of extended spectrum in selective chromogenic agar was analyzed The results ofdetection of beta-lactamases of extended spectrum was compared with "double disc" technique. The smears from mucous membrane of guttur and rectum from patients were analyzed in parallel on solid growth agar (Endo or Mac Conkey) and on selective agar CHROMagartm ESBL (CHROMagar France). The production of beta-lactamases of extended spectrum was confirmed using "double discs" technique. To exclude hyper-production of ampC beta-lactamases E-test was applied containing cefotetan and cefotetan with cloxacillin. The sampling consisted of 1552 samples from patients. The study permitted to isolate 1243 strains of enterobacteria on agar Endo or Mac Conkey and 409 strains of enterobacteria on selective agar CHROMagartm ESBL (Escherichia coli n = 226, Klebsiella pneumoniae n = 105, enterobacter spp. n = 35, Citrobacter spp. n = 21, others n = 22). The application of "double discs" technique confirmed production of beta-lactamases of extended spectrum in 386 (94%) out of 409 strains isolated on agar CHROMagartm ESBL. In 23 (6%) of strains no confirmation was established and hyper-production of ampC of beta-lactamases was established 15 out of total. Additionally, 8 were sensitive to cephalosporin of third generation. All enterobacteria isolated on agar Endo or Mac Conkey also were tested by "double discs" technique. Overall, 394 strains of enterobacteria with production of beta-lactamases of extended spectrum were obtained. On all agars (agar Endo or Mac Conkey and CHROMagartm ESBL)--263 (67%) strains; only on CHROMagartm ESBL--123 (31%) and only on agar Endo or Mac Conkey--8 (2%) (p < 0.0001). The sensitivity of selective agar CHROMagartm ESBL made up to 98% and specificity--97%. The resolution about detection of enterobacteria producing beta-lactamases of extended spectrum were submitted to clinic in 18-24 hours after arrival

  6. [THE APPLICATION OF SELECTIVE CHROMOGENIC AGAR FOR DETECTING ENTEROBACTERIA WITH PRODUCTION OF BETA-LACTAMASES].

    PubMed

    Korobova, A G; Frolova, L N; Kliasova, G A

    2015-11-01

    The detection of enterobacteria with production of beta-lactamases of extended spectrum in selective chromogenic agar was analyzed The results ofdetection of beta-lactamases of extended spectrum was compared with "double disc" technique. The smears from mucous membrane of guttur and rectum from patients were analyzed in parallel on solid growth agar (Endo or Mac Conkey) and on selective agar CHROMagartm ESBL (CHROMagar France). The production of beta-lactamases of extended spectrum was confirmed using "double discs" technique. To exclude hyper-production of ampC beta-lactamases E-test was applied containing cefotetan and cefotetan with cloxacillin. The sampling consisted of 1552 samples from patients. The study permitted to isolate 1243 strains of enterobacteria on agar Endo or Mac Conkey and 409 strains of enterobacteria on selective agar CHROMagartm ESBL (Escherichia coli n = 226, Klebsiella pneumoniae n = 105, enterobacter spp. n = 35, Citrobacter spp. n = 21, others n = 22). The application of "double discs" technique confirmed production of beta-lactamases of extended spectrum in 386 (94%) out of 409 strains isolated on agar CHROMagartm ESBL. In 23 (6%) of strains no confirmation was established and hyper-production of ampC of beta-lactamases was established 15 out of total. Additionally, 8 were sensitive to cephalosporin of third generation. All enterobacteria isolated on agar Endo or Mac Conkey also were tested by "double discs" technique. Overall, 394 strains of enterobacteria with production of beta-lactamases of extended spectrum were obtained. On all agars (agar Endo or Mac Conkey and CHROMagartm ESBL)--263 (67%) strains; only on CHROMagartm ESBL--123 (31%) and only on agar Endo or Mac Conkey--8 (2%) (p < 0.0001). The sensitivity of selective agar CHROMagartm ESBL made up to 98% and specificity--97%. The resolution about detection of enterobacteria producing beta-lactamases of extended spectrum were submitted to clinic in 18-24 hours after arrival

  7. 40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... interval. You may use the difference between a diluted exhaust flow meter and a dilution air meter to.... We recommend that you use a diluted exhaust flow meter that meets the specifications in Table 1 of... verification in § 1065.307 and the calibration and verifications in § 1065.340 and § 1065.341. You may use...

  8. 40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... interval. You may use the difference between a diluted exhaust flow meter and a dilution air meter to.... We recommend that you use a diluted exhaust flow meter that meets the specifications in Table 1 of... verification in § 1065.307 and the calibration and verifications in § 1065.340 and § 1065.341. You may use...

  9. 40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... interval. You may use the difference between a diluted exhaust flow meter and a dilution air meter to.... We recommend that you use a diluted exhaust flow meter that meets the specifications in Table 1 of... verification in § 1065.307 and the calibration and verifications in § 1065.340 and § 1065.341. You may use...

  10. 40 CFR 1065.240 - Dilution air and diluted exhaust flow meters.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... interval. You may use the difference between a diluted exhaust flow meter and a dilution air meter to.... We recommend that you use a diluted exhaust flow meter that meets the specifications in Table 1 of... verification in § 1065.307 and the calibration and verifications in § 1065.340 and § 1065.341. You may use...

  11. Isolation of Typical Marine Bacteria by Dilution Culture: Growth, Maintenance, and Characteristics of Isolates under Laboratory Conditions

    PubMed Central

    Schut, Frits; de Vries, Egbert J.; Gottschal, Jan C.; Robertson, Betsy R.; Harder, Wim; Prins, Rudolf A.; Button, Don K.

    1993-01-01

    Marine bacteria in Resurrection Bay near Seward, Alaska, and in the central North Sea off the Dutch coast were cultured in filtered autoclaved seawater following dilution to extinction. The populations present before dilution varied from 0.11 × 109 to 1.07 × 109 cells per liter. The mean cell volume varied between 0.042 and 0.074 μm3, and the mean apparent DNA content of the cells ranged from 2.5 to 4.7 fg of DNA per cell. All three parameters were determined by high-resolution flow cytometry. All 37 strains that were obtained from very high dilutions of Resurrection Bay and North Sea samples represented facultatively oligotrophic bacteria. However, 15 of these isolates were eventually obtained from dilution cultures that could initially be cultured only on very low-nutrient media and that could initially not form visible colonies on any of the agar media tested, indicating that these cultures contained obligately oligotrophic bacteria. It was concluded that the cells in these 15 dilution cultures had adapted to growth under laboratory conditions after several months of nutrient deprivation prior to isolation. From the North Sea experiment, it was concluded that the contribution of facultative oligotrophs and eutrophs to the total population was less than 1% and that while more than half of the population behaved as obligately oligotrophic bacteria upon first cultivation in the dilution culture media, around 50% could not be cultured at all. During one of the Resurrection Bay experiments, 53% of the dilution cultures obtained from samples diluted more than 2.5 × 105 times consisted of such obligate oligotrophs. These cultures invariably harbored a small rod-shaped bacterium with a mean cell volume of 0.05 to 0.06 μm3 and an apparent DNA content of 1 to 1.5 fg per cell. This cell type had the dimensions of ultramicrobacteria. Isolates of these ultramicrobacterial cultures that were eventually obtained on relatively high-nutrient agar plates were, with respect

  12. Electrocoalescence based serial dilution of microfluidic droplets.

    PubMed

    Bhattacharjee, Biddut; Vanapalli, Siva A

    2014-07-01

    Dilution of microfluidic droplets where the concentration of a reagent is incrementally varied is a key operation in drop-based biological analysis. Here, we present an electrocoalescence based dilution scheme for droplets based on merging between moving and parked drops. We study the effects of fluidic and electrical parameters on the dilution process. Highly consistent coalescence and fine resolution in dilution factor are achieved with an AC signal as low as 10 V even though the electrodes are separated from the fluidic channel by insulator. We find that the amount of material exchange between the droplets per coalescence event is high for low capillary number. We also observe different types of coalescence depending on the flow and electrical parameters and discuss their influence on the rate of dilution. Overall, we find the key parameter governing the rate of dilution is the duration of coalescence between the moving and parked drop. The proposed design is simple incorporating the channel electrodes in the same layer as that of the fluidic channels. Our approach allows on-demand and controlled dilution of droplets and is simple enough to be useful for assays that require serial dilutions. The approach can also be useful for applications where there is a need to replace or wash fluid from stored drops.

  13. Electrocoalescence based serial dilution of microfluidic droplets

    PubMed Central

    Bhattacharjee, Biddut; Vanapalli, Siva A.

    2014-01-01

    Dilution of microfluidic droplets where the concentration of a reagent is incrementally varied is a key operation in drop-based biological analysis. Here, we present an electrocoalescence based dilution scheme for droplets based on merging between moving and parked drops. We study the effects of fluidic and electrical parameters on the dilution process. Highly consistent coalescence and fine resolution in dilution factor are achieved with an AC signal as low as 10 V even though the electrodes are separated from the fluidic channel by insulator. We find that the amount of material exchange between the droplets per coalescence event is high for low capillary number. We also observe different types of coalescence depending on the flow and electrical parameters and discuss their influence on the rate of dilution. Overall, we find the key parameter governing the rate of dilution is the duration of coalescence between the moving and parked drop. The proposed design is simple incorporating the channel electrodes in the same layer as that of the fluidic channels. Our approach allows on-demand and controlled dilution of droplets and is simple enough to be useful for assays that require serial dilutions. The approach can also be useful for applications where there is a need to replace or wash fluid from stored drops. PMID:25379096

  14. Testing susceptibility of multidrug-resistant Mycobacterium tuberculosis to second-line drugs by use of blood agar.

    PubMed

    Satana, Dilek; Coban, Ahmet Yilmaz; Uzun, Meltem

    2010-11-01

    In this study, the susceptibilities of 35 multidrug-resistant (MDR) Mycobacterium tuberculosis clinical isolates to second-line drugs, including kanamycin (KM), rifabutin (RBU), ofloxacin (OFX), p-aminosalicylic acid (PAS), capreomycin (CAP), clofazimine (CFM), and ethionamide (ETH), were investigated on blood agar according to CLSI recommendations. Compared with the results of the Bactec 460 TB system, agreement was 100, 100, 97, 100, 100, 100, and 86% for KM, RBU, OFX, PAS, CAP, CFM, and ETH, respectively. Compared with the results of the proportion method, agreement was 100, 100, 97, 100, 97, 100, and 77% for KM, RBU, OFX, PAS, CAP, CFM, and ETH, respectively.

  15. A new chromogenic agar medium for detection of potentially virulent Yersinia enterocolitica.

    PubMed

    Weagant, Stephen D

    2008-02-01

    Several outbreaks of foodborne yersiniosis have been documented and this disease continues to be source of infections transmitted through foods. The selective agars most commonly used to isolate Yersinia enterocolitica in clinical, food and environmental samples, cefsulodin-irgasan-novobiocin (CIN) and MacConkey (MAC) agars, lack the ability to differentiate potentially virulent Y. enterocolitica from other Yersinia that may be present as well as some other bacterial spp. This study proposes the use of an agar medium, Y. enterocolitica chromogenic medium (YeCM), for isolation of potentially virulent Y. enterocolitica. This agar contains cellobiose as the fermentable sugar, a chromogenic substrate and selective inhibitors for suppression of colony formation by many competing bacteria. All strains of potentially virulent Yersinia of biotypes 1B, and biotypes 2-5 formed convex, red bulls-eye colonies on YeCM that were very similar to those described for CIN agar. However, Y. enterocolitica biotype 1A and other related Yersinia formed colonies that were purple/blue on YeCM while they formed typical red bulls-eye colonies on CIN agar. When a mixture of potentially virulent Y. enterocolitica biotype 1B, Y. enterocolitica biotype 1A and 5 other bacterial species was used to artificially contaminate tofu and then spread-plated on three selective agars, Y. enterocolitica biotype 1B colonies were easily distinguished from other strains on YeCM. However, Y. enterocolitica biotype 1B colonies were indistinguishable from many other colonies on CIN and only distinguishable from those of C. freundii on MAC. When colonies were picked and identified from these agars, typical colonies from YeCM were confirmed only as Y. enterocolitica biotype 1B. Typical colonies on CIN and MAC were found to belong to several competing species and biotypes.

  16. Genome Sequence of the Agar-Degrading Marine Bacterium Alteromonadaceae sp. Strain G7

    PubMed Central

    Kwak, Min-Jung; Song, Ju Yeon; Kim, Byung Kwon; Chi, Won-Jae; Kwon, Soon-Kyeong; Choi, Soobeom; Chang, Yong-Keun

    2012-01-01

    Here, we present the high-quality draft genome sequence of the agar-degrading marine gammaproteobacterium Alteromonadaceae sp. strain G7, which was isolated from coastal seawater to be utilized as a bioresource for production of agar-derived biofuels. The 3.91-Mb genome contains a number of genes encoding algal polysaccharide-degrading enzymes such as agarases and sulfatases. PMID:23209220

  17. Detection of Activity Responsible for Induction of the Agrobacterium tumefaciens Virulence Genes in Bacteriological Agar.

    PubMed

    Loubens, I; Chilton, W S; Dion, P

    1997-11-01

    Agrobacterium tumefaciens C58 grown on acidic medium containing glucose and solidified with bacteriological agar expressed a virB::lacZ fusion. No expression of this fusion was observed on a similar medium which was solidified with purified agarose. The fraction from bacteriological agar which was responsible for vir gene induction was extracted with methanol and partially purified by preparative thin-layer chromatography. PMID:16535740

  18. Evaluation of the Oxoid Brilliance™ CRE Agar for the detection of carbapenemase-producing Enterobacteriaceae.

    PubMed

    Cohen Stuart, J; Voets, G; Rottier, W; Voskuil, S; Scharringa, J; Van Dijk, K; Fluit, A C; Leverstein-Van Hall, M

    2013-11-01

    The adequate detection of carbapenemase-producing Enterobacteriaceae (CPE) is essential for adequate antibiotic therapy and for infection control purposes, especially in an outbreak setting. Selective agars play an important role in the detection of CPE. The Oxoid Brilliance™ CRE Agar (Thermo Fisher Scientific) was evaluated for the detection of CPE using 255 non-repetitive Enterobacteriaceae isolates, including 95 CPE (36 KPC, 4 KPC plus VIM, 4 NDM, 6 GIM, 20 VIM, and 25 OXA-48-producing isolates). The sensitivity of the CRE agar for the detection of CPE was 94 % (89/95), but differed per carbapenemase gene (100 % for KPC, NDM, and GIM, 90 % for VIM, and 84 % for OXA-48-producing isolates). The specificity of the CRE agar was 71 %, due to the growth of AmpC- and/or ESBL-producing isolates. The CRE agar is a sensitive tool for the detection of KPC and metallo-carbapenemase-producing Enterobacteriaceae, although the detection of OXA-48 producers is less optimal. The relatively low specificity requires confirmation of carbapenemase production for isolates recovered from the CRE agar.

  19. Entrapment of α-Amylase in Agar Beads for Biocatalysis of Macromolecular Substrate

    PubMed Central

    Sharma, Manu; Sharma, Vinay; Majumdar, Dipak K.

    2014-01-01

    Attempts have been made to optimize immobilization parameters, catalytic property, and stability of immobilized α-amylase in agar. The work compares natural entrapment efficiency of agar with the ionotropically cross-linked agar hydrogel, with the advantage of easy scale-up and cost and time effectiveness. Beads prepared with 3% (w/v) agar and 75 mM calcium chloride and hardened for 20 minutes were selected for further studies on the basis of entrapment efficiency (80%) and physical stability. Following entrapment, pH and temperature optima of enzyme were shifted from 6 to 6.5 and 50 to 55°C, respectively. Michaelis constant (Km) for both free and entrapped enzymes remained the same (0.83%) suggesting no change in substrate affinity. However, Vmax⁡ of entrapped enzyme decreased ~37.5-fold. The midpoint of thermal inactivation for entrapped enzyme increased by 8 ± 1°C implying its higher thermal stability. The entrapped enzyme in calcium agar bead had an Ea value of 27.49 kcal/mol compared to 17.6 kcal/mol for free enzyme indicating increased stability on entrapment. Half-life of enzyme increased ~2.2 times after entrapment in calcium agar at 60°C indicating stabilization of enzyme. The reusability of beads was size dependent. Beads with diameter <710 μm were stable and could be reused for 6 cycles with ~22% loss in activity. PMID:27382608

  20. Complex impedance and conductivity of agar-based ion-conducting polymer electrolytes

    NASA Astrophysics Data System (ADS)

    Nwanya, A. C.; Amaechi, C. I.; Udounwa, A. E.; Osuji, R. U.; Maaza, M.; Ezema, F. I.

    2015-04-01

    Agar-based electrolyte standing films with different salts and weak acids as ion and proton conductors were prepared and characterized by X-ray diffraction, UV-visible spectrophotometry, photoluminescence emission spectroscopy and electrochemical impedance spectroscopy. The salts used are lithium perchlorate (LiClO4) and potassium perchlorate (KClO4), while the weak acids used are acetic acid (CH3COOH) and lactic acid (C3H6O3). The values of the ion conductivity obtained for the agar-based polymer films are 6.54 × 10-8, 9.12 × 10-8, 3.53 × 10-8, 2.24 × 10-8 S/cm for the agar/acetic acid, agar/lactic acid, agar/LiClO4 and agar/KClO4 polymer films, respectively. As a function of temperature, the ion conductivity exhibits an Arrhenius behavior and the estimated activation energy is ≈0.1 eV for all the samples. The samples depicted high values of dielectric permittivity toward low frequencies which is due mostly to electrode polarization effect. The samples showed very high transparency (85-98 %) in the visible region, and this high transparency is one of the major requirements for application in electrochromic devices (ECD). The values of conductivity and activation energy obtained indicate that the electrolytes are good materials for application in ECD.

  1. [GROWTH OF MICROMYCETES FROM DIFFERENT ECOLOGICAL NICHES ON AGAR NUTRIENT MEDIA].

    PubMed

    Kurchenko, I M; Yurieva, E M; Voychuk, S I

    2015-01-01

    Radial growth rate of (K(r)) 153 strains 6 species of micromycetes from different ecological niches was studied on 7 agar media: three standard (malt extract agar, potato-dextrose agar, Czapek's agar), and on agar media with plant polymers (carboxymethylcellulose, xylan, soluble starch and apple pectin). Endophytic and plant pathogenic strains (biotrophs) of all studied species did not differ significantly in their ability to grow on nutrient media of different composition--average values of K(r) for these two groups were the same (0,200 and 0,199 mm/h, respectively). Soil micromycetes (saprophytes) characterized by the lowest average growth rate (0,169 mm/h) and significantly differed from the endophytic and plant pathogenic ones. Average of the radial growth rates of studied microscopic fungi were higher on standard nutrient media than with plant polymers ones. Growth parameters of endophytes and plant pathogens of all studied species on various agar media differed from the soil strains. High growth rate of endophytic and plant pathogenic strains of Fusarium poae, Alternaria alternata and Ceratocystis sp. provides them the rapid colonization of plants. Penicillium funiculosum strains equally can exist as saprophytes in soil and as endophytic plant symbionts. A wide range of K(r) variation of endophytic dark pigmented Mycelia sterilia indicates the presence in this group of different species of micromycetes, which have no sporulation.

  2. Broth versus solid agar culture of swab samples of cadaveric allograft musculoskeletal tissue.

    PubMed

    Varettas, Kerry

    2013-12-01

    As part of the donor assessment protocol, bioburden assessment must be performed on allograft musculoskeletal tissue samples collected at the time of tissue retrieval. Swab samples of musculoskeletal tissue allografts from cadaveric donors are received at the microbiology department of the South Eastern Area Laboratory Services (Australia) to determine the presence of bacteria and fungi. This study will review the isolation rate of organisms from solid agar and broth culture of swab samples of cadaveric allograft musculoskeletal tissue over a 6-year period, 2006-2011. Swabs were inoculated onto horse blood agar (anaerobic, 35 °C) and chocolate agar (CO2, 35 °C) and then placed into a cooked meat broth (aerobic, 35 °C). A total of 1,912 swabs from 389 donors were received during the study period. 557 (29.1 %) swabs were culture positive with the isolation of 713 organisms, 249 (34.9 %) from solid agar culture and an additional 464 (65.1 %) from broth culture only. This study has shown that the broth culture of cadaveric allograft musculoskeletal swab samples recovered a greater amount of organisms than solid agar culture. Isolates such as Clostridium species and Staphylococcus aureus would not have been isolated from solid agar culture alone. Broth culture is an essential part of the bioburden assessment protocol of swab samples of cadaveric allograft musculoskeletal tissue in this laboratory.

  3. Susceptibilities of Mycobacterium tuberculosis to isoniazid and rifampin on blood agar.

    PubMed

    Coban, Ahmet Yilmaz; Bilgin, Kemal; Uzun, Meltem; Tasdelen Fisgin, Nuriye; Akgunes, Alper; Cihan, Cigdem Cekic; Birinci, Asuman; Durupinar, Belma

    2005-04-01

    In this study, blood agar was used instead of 7H10 agar for the susceptibility testing of 34 clinical isolates of Mycobacterium tuberculosis to isoniazid (INH) and rifampin (RIF) in accordance with the NCCLS. The BACTEC 460 TB system (Becton Dickinson, Sparks, Md.) was used as a "gold standard." Results for both media were in agreement for RIF and INH at 100 and 94.1%, respectively. For INH, the specificity, sensitivity, positive predictive value, and negative predictive value were found to be 71.4, 100, 93.1, and 100%, respectively, while these values were 100% for RIF. In addition, the results of the susceptibility test performed with blood agar were obtained on day 14 of incubation. In conclusion, results were obtained much earlier with blood agar (2 weeks) than with 7H10 agar (3 weeks), and the results of this study suggest that blood agar may be used as an alternative medium for the susceptibility testing of M. tuberculosis to INH and RIF.

  4. Optimization of modified Middlebrook 7H11 agar for isolation of Mycobacterium bovis from raw milk cheese.

    PubMed

    Forgrave, R; Donaghy, J A; Fisher, A; Rowe, M T

    2014-10-01

    Reports have highlighted the absence of contemporary peer reviewed publications pertaining to Mycobacterium bovis culture from raw milk and cheese. By replicating traditional methods, cheese-making methodology and equipment were devised to produce Cheddar (n = 6) and Caerphilly (n = 3) artificially contaminated with M. bovis (three genotypes) under stringent laboratory-containment guidelines for handling hazardous microbiological material. Middlebrook 7H11, modified for M. bovis isolation, was assessed for capacity to enumerate M. bovis despite changing cheese microflora and prolonged M. bovis exposure to the cheese matrix using maturing cheese test portions (n = 63; up to 16 weeks). Malachite green (MG) containing media isolated M. bovis at significantly (P < 0·05) lower levels than unmodified Middlebrook 7H11 agar despite MG being a common adjunct of Middlebrook 7H11 agar modified for M. bovis growth. Subsequently, a selective MG-free Middlebrook 7H11 agar modified using haemolysed red cells and calf serum was demonstrated as the best performing (P < 0·05) medium for recovery of M. bovis from typical UK cheese types, Cheddar and Caerphilly. Significance and impact of the study: Following increased M. bovis infection of UK cattle, the risk posed to consumers from consumption of unpasteurized milk and dairy products has changed. Furthermore, published methods for the culture and molecular detection of M. bovis in raw milk products are limited. Cheese-making protocols and M. bovis culture media reported here provide tools for further investigation of M. bovis survival during all stages of cheese manufacture and could inform future assessment of the risk to consumers from M. bovis contamination of unpasteurized dairy products.

  5. Optimization of modified Middlebrook 7H11 agar for isolation of Mycobacterium bovis from raw milk cheese.

    PubMed

    Forgrave, R; Donaghy, J A; Fisher, A; Rowe, M T

    2014-10-01

    Reports have highlighted the absence of contemporary peer reviewed publications pertaining to Mycobacterium bovis culture from raw milk and cheese. By replicating traditional methods, cheese-making methodology and equipment were devised to produce Cheddar (n = 6) and Caerphilly (n = 3) artificially contaminated with M. bovis (three genotypes) under stringent laboratory-containment guidelines for handling hazardous microbiological material. Middlebrook 7H11, modified for M. bovis isolation, was assessed for capacity to enumerate M. bovis despite changing cheese microflora and prolonged M. bovis exposure to the cheese matrix using maturing cheese test portions (n = 63; up to 16 weeks). Malachite green (MG) containing media isolated M. bovis at significantly (P < 0·05) lower levels than unmodified Middlebrook 7H11 agar despite MG being a common adjunct of Middlebrook 7H11 agar modified for M. bovis growth. Subsequently, a selective MG-free Middlebrook 7H11 agar modified using haemolysed red cells and calf serum was demonstrated as the best performing (P < 0·05) medium for recovery of M. bovis from typical UK cheese types, Cheddar and Caerphilly. Significance and impact of the study: Following increased M. bovis infection of UK cattle, the risk posed to consumers from consumption of unpasteurized milk and dairy products has changed. Furthermore, published methods for the culture and molecular detection of M. bovis in raw milk products are limited. Cheese-making protocols and M. bovis culture media reported here provide tools for further investigation of M. bovis survival during all stages of cheese manufacture and could inform future assessment of the risk to consumers from M. bovis contamination of unpasteurized dairy products. PMID:24888395

  6. Comparative in vitro activity of Meropenem, Imipenem and Piperacillin/tazobactam against 1071 clinical isolates using 2 different methods: a French multicentre study

    PubMed Central

    2010-01-01

    Background Meropenem is a carbapenem that has an excellent activity against many gram-positive and gram-negative aerobic, facultative, and anaerobic bacteria. The major objective of the present study was to assess the in vitro activity of meropenem compared to imipenem and piperacillin/tazobactam, against 1071 non-repetitive isolates collected from patients with bacteremia (55%), pneumonia (29%), peritonitis (12%) and wound infections (3%), in 15 French hospitals in 2006. The secondary aim of the study was to compare the results of routinely testings and those obtained by a referent laboratory. Method Susceptibility testing and Minimum Inhibitory Concentrations (MICs) of meropenem, imipenem and piperacillin/tazobactam were determined locally by Etest method. Susceptibility to meropenem was confirmed at a central laboratory by disc diffusion method and MICs determined by agar dilution method for meropenem, imipenem and piperacillin/tazobactam. Results Cumulative susceptibility rates against Escherichia coli were, meropenem and imipenem: 100% and piperacillin/tazobactam: 90%. Against other Enterobacteriaceae, the rates were meropenem: 99%, imipenem: 98% and piperacillin/tazobactam: 90%. All Staphylococci, Streptococci and anaerobes were susceptible to the three antibiotics. Against non fermeters, meropenem was active on 84-94% of the strains, imipenem on 84-98% of the strains and piperacillin/tazobactam on 90-100% of the strains. Conclusions Compared to imipenem, meropenem displays lower MICs against Enterobacteriaceae, Escherichia coli and Pseudomonas aeruginosa. Except for non fermenters, MICs90 of carbapenems were <4 mg/L. Piperacillin/tazobactam was less active against Enterobacteriaceae and Acinetobacter but not P. aeruginosa. Some discrepancies were noted between MICs determined by Etest accross centres and MICs determined by agar dilution method at the central laboratory. Discrepancies were more common for imipenem testing and more frequently related to a few

  7. Chromogenic assay of human coagulation factor VIII: statistical comparison of 2 working dilution procedures.

    PubMed

    Alonso, C; Gonzalez, A; Frutos, G

    2005-08-01

    The effect of 2 different practices for preparation of working dilutions in the chromogenic substrate method for potency assay of factor VIII was evaluated. In this study the potency of several concentrate materials was shown to be statistically equivalent, whether performing the assay with independent or serial working dilutions.

  8. Preparation, Optimization, and Screening of the Effect of Processing Variables on Agar Nanospheres Loaded with Bupropion HCl by a D-Optimal Design.

    PubMed

    Varshosaz, Jaleh; Zaki, Mohammad Reza; Minaiyan, Mohsen; Banoozadeh, Jaafar

    2015-01-01

    Bupropion is an atypical antidepressant drug. Fluctuating in its serum levels following oral administration of immediate release dosage forms leads to occasional seizure. The aim of the present work was designing of sustained release bupropion HCl nanospheres suited for pulmonary delivery. Agar nanospheres were prepared by transferring the w/o emulsion to solid in oil (s/o) suspension. Calcium chloride was used as cross-linking agent and hydroxypropyl β-cyclodextrin (HPβCD) was used as permeability enhancer. A response surface D-optimal design was used for optimization of nanospheres. Independent factors included in the design were calcium chloride percent, speed of homogenization, agar percent, and HPβCD percent. Optimum condition was predicted to be achieved when the calcium chloride was set at 7.19%, homogenization speed at 8500 rpm, agar content at 2%, and HPβCD at 0.12%. The optimized nanoparticles showed particle size of 587 nm, zeta potential of -30.9 mV, drug loading efficiency of 38.6%, and release efficiency of 51% until 5 h. The nanospheres showed high degree of bioadhesiveness. D-optimal response surface method is a satisfactory design to optimize the fabrication of bupropion HCl loaded agar nanospheres and these nanospheres can be successively exploited to deliver bupropion in a controlled manner for a sufficiently extended period.

  9. Preparation, Optimization, and Screening of the Effect of Processing Variables on Agar Nanospheres Loaded with Bupropion HCl by a D-Optimal Design

    PubMed Central

    Varshosaz, Jaleh; Zaki, Mohammad Reza; Minaiyan, Mohsen; Banoozadeh, Jaafar

    2015-01-01

    Bupropion is an atypical antidepressant drug. Fluctuating in its serum levels following oral administration of immediate release dosage forms leads to occasional seizure. The aim of the present work was designing of sustained release bupropion HCl nanospheres suited for pulmonary delivery. Agar nanospheres were prepared by transferring the w/o emulsion to solid in oil (s/o) suspension. Calcium chloride was used as cross-linking agent and hydroxypropyl β-cyclodextrin (HPβCD) was used as permeability enhancer. A response surface D-optimal design was used for optimization of nanospheres. Independent factors included in the design were calcium chloride percent, speed of homogenization, agar percent, and HPβCD percent. Optimum condition was predicted to be achieved when the calcium chloride was set at 7.19%, homogenization speed at 8500 rpm, agar content at 2%, and HPβCD at 0.12%. The optimized nanoparticles showed particle size of 587 nm, zeta potential of −30.9 mV, drug loading efficiency of 38.6%, and release efficiency of 51% until 5 h. The nanospheres showed high degree of bioadhesiveness. D-optimal response surface method is a satisfactory design to optimize the fabrication of bupropion HCl loaded agar nanospheres and these nanospheres can be successively exploited to deliver bupropion in a controlled manner for a sufficiently extended period. PMID:26090423

  10. A comparison of the performance of commercially available chromogenic agars for the isolation and presumptive identification of organisms from urine

    PubMed Central

    Fallon, D; Ackland, G; Andrews, N; Frodsham, D; Howe, S; Howells, K; Nye, K J; Warren, R E

    2003-01-01

    Aims: To compare four media—UTI medium, BBL CHROMagar, CPS ID2, and Harlequin CLED—using a collection of fully characterised organisms and subsequent “field trial”. Methods: Seven hundred and eighty seven fully characterised isolates (730 Gram negative bacteria, 47 Gram positive bacteria, and 10 yeasts) were used to test for accuracy of organism identification. To assess isolation rates and ability to detect mixed cultures, 1435 urine samples were cultured in the three best performing chromogenic media (UTI medium, BBL CHROMagar, and CPS ID2) and CLED. Results: The chromogenic agars differed in their accuracy of identification, with BBL CHROMagar performing best and Harlequin CLED performing least well. Similarly, BBL CHROMagar achieved a higher overall isolation rate than UTI medium and CPS ID2. When mixed growth was defined as greater than two organism types, BBL CHROMagar detected more mixed cultures than did UTI medium and CPS ID2, although the differences were not significant. When mixed growth was defined as greater than one organism type the increased number of mixed growths detected by BBL CHROMagar became significant, largely because of differences in enterococcal isolation rates. Conclusion: The use of BBL CHROMagar, UTI medium, or CPS ID2 chromogenic agar as a replacement for CLED agar would improve the detection rate of contaminated urine samples. Enhanced identification helps to distinguish different species, facilitating the monitoring of bacterial resistance in support of the national antibiotic strategy. BBL CHROMagar gave the highest overall organism recovery rates, greatest ability to detect mixed cultures, and the most accurate identification of organisms. PMID:12890812

  11. Injection of Acanthaster planci with thiosulfate-citrate-bile-sucrose agar (TCBS). II. Histopathological changes.

    PubMed

    Rivera-Posada, J A; Pratchett, M; Owens, L

    2011-12-01

    We assessed histological changes in the tissues of the crown-of-thorns starfish Acanthaster planci (COTS) after injection of thiosulfate-citrate-bile-sucrose agar (TCBS) which was used as a disease inducer (potential outbreak control method), by conventional and scanning electron microscopy. Digestive glands were processed and stained with hematoxylin and eosin to describe the histological architecture of the intestinal epithelium. Subsequently comparison of healthy versus infected tissues and Gram stains were carried out to confirm bacterial occurrence on infected tissues, characterize the structural changes induced by bacterial communities in COTS tissues, and to determine if the histopathological changes of intestinal tissues were consistent with vibrio infection. TCBS injections induced marked epithelial desquamation, hypertrophy and hypersecretion of glandular cells, epithelial cell destruction, pyknosis, reduction of thickness and disorganization of connective tissue and associated nerve plexus, presence of bacterial colonies, irregular eosinophilic foci in glandular cells, brush border disruption, atrophy and detachment of intestinal microvilli and cell debris in the lumen. All these changes were attributed to a fulminating systemic dysbiosis and were consistent with vibrio infections.

  12. Development and validation of a microbiological agar assay for determination of orbifloxacin in pharmaceutical preparations.

    PubMed

    Cazedey, Edith C L; Salgado, Hérida R N

    2011-01-01

    Orbifloxacin is a fluoroquinolone with broad-spectrum antimicrobial activity, and belongs to the third generation of quinolones. Regarding the quality control of medicines, a validated microbiological assay for determination of orbifloxacin in pharmaceutical formulations has not as yet been reported. For this purpose, this paper reports the development and validation of a simple, sensitive, accurate and reproducible agar diffusion method to quantify orbifloxacin in tablet formulations. The assay is based on the inhibitory effect of orbifloxacin upon the strain of Staphylococcus aureus ATCC 25923 used as test microorganism. The results were treated statistically by analysis of variance and were found to be linear (r = 0.9992) in the selected range of 16.0-64.0 μg/mL, precise with relative standard deviation (RSD) of repeatability intraday = 2.88%, intermediate precision RSD = 3.33%, and accurate (100.31%). The results demonstrated the validity of the proposed bioassay, which allows reliable orbifloxacin quantitation in pharmaceutical samples and therefore can be used as a useful alternative methodology for the routine quality control of this medicine.

  13. Development and Validation of a Microbiological Agar Assay for Determination of Orbifloxacin in Pharmaceutical Preparation

    PubMed Central

    Cazedey, Edith C. L.; Salgado, Hérida R. N.

    2011-01-01

    Orbifloxacin is a fluoroquinolone with broad-spectrum antimicrobial activity, and belongs to the third generation of quinolones. Regarding the quality control of medicines, a validated microbiological assay for determination of orbifloxacin in pharmaceutical formulations has not as yet been reported. For this purpose, this paper reports the development and validation of a simple, sensitive, accurate and reproducible agar diffusion method to quantify orbifloxacin in tablet formulations. The assay is based on the inhibitory effect of orbifloxacin upon the strain of Staphylococcus aureus ATCC 25923 used as test microorganism. The results were treated statistically by analysis of variance and were found to be linear (r = 0.9992) in the selected range of 16.0–64.0 μg/mL, precise with relative standard deviation (RSD) of repeatability intraday = 2.88%, intermediate precision RSD = 3.33%, and accurate (100.31%). The results demonstrated the validity of the proposed bioassay, which allows reliable orbifloxacin quantitation in pharmaceutical samples and therefore can be used as a useful alternative methodology for the routine quality control of this medicine. PMID:24310597

  14. Synthesis and characterization of high-surface-area millimeter-sized silica beads with hierarchical multi-modal pore structure by the addition of agar

    SciTech Connect

    Han, Yosep; Choi, Junhyun; Tong, Meiping; Kim, Hyunjung

    2014-04-01

    Millimeter-sized spherical silica foams (SSFs) with hierarchical multi-modal pore structure featuring high specific surface area and ordered mesoporous frameworks were successfully prepared using aqueous agar addition, foaming and drop-in-oil processes. The pore-related properties of the prepared spherical silica (SSs) and SSFs were systematically characterized by field emission-scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), small-angle X-ray diffraction (SAXRD), Hg intrusion porosimetry, and N{sub 2} adsorption–desorption isotherm measurements. Improvements in the BET surface area and total pore volume were observed at 504 m{sup 2} g{sup −1} and 5.45 cm{sup 3} g{sup −1}, respectively, after an agar addition and foaming process. Despite the increase in the BET surface area, the mesopore wall thickness and the pore size of the mesopores generated from the block copolymer with agar addition were unchanged based on the SAXRD, TEM, and BJH methods. The SSFs prepared in the present study were confirmed to have improved BET surface area and micropore volume through the agar loading, and to exhibit interconnected 3-dimensional network macropore structure leading to the enhancement of total porosity and BET surface area via the foaming process. - Highlights: • Millimeter-sized spherical silica foams (SSFs) are successfully prepared. • SSFs exhibit high BET surface area and ordered hierarchical pore structure. • Agar addition improves BET surface area and micropore volume of SSFs. • Foaming process generates interconnected 3-D network macropore structure of SSFs.

  15. Helium Dilution Cryocooler for Space Applications

    NASA Technical Reports Server (NTRS)

    Roach, Pat; Hogan, Robert (Technical Monitor)

    2001-01-01

    NASA's New Millenium Program Space Technology presents the Helium Dilution Cryocooler for Space Applications. The topics include: 1) Capability; 2) Applications; and 3) Advantages. This paper is in viewgraph form.

  16. Improvement of Karmali agar by addition of polymyxin B for the detection of Campylobacter jejuni and C. coli in whole-chicken carcass rinse.

    PubMed

    Chon, Jung-Whan; Kim, Hyunsook; Yim, Jin-Hyeok; Song, Kwang-Young; Moon, Jin-San; Kim, Young-Jo; Seo, Kun-Ho

    2013-05-01

    The Karmali agar was modified by supplementation with a high concentration of polymyxin B. The goal of the study was to evaluate the effect of a high concentration of polymyxin B on the ability and selectivity of the modified Karmali agar to isolate Campylobacter jejuni and Campylobacter coli from whole chicken carcass rinse. A total of 80 whole chickens were rinsed with 400 mL of buffer peptone water. The rinsed samples were incubated with 2× blood-free modified Bolton enrichment broth for 48 h, and then streaked onto unmodified Karmali agar and modified Karmali agar supplemented with 100000 IU/L polymixin B (P-Karmali agar). The suspected colonies were finally confirmed by colony PCR. The P-Karmali agar exhibited a significantly better (P < 0.05) isolation rate than the unmodified Karmali agar (P-Karmali agar, 73.8%; unmodified Karmali agar, 33.8%). Moreover, the selectivity of the P-Karmali agar was also better (P < 0.05) than that of the other selective agar when comparing the number of contaminated plates (P-Karmali agar, 68.8%; unmodified Karmali agar, 87.5%) and growth index of competing flora (P-Karmali agar, 1.4; unmodified Karmali agar, 2.7). The improved selective agar excluded competing flora resistant to antibiotic agents in unmodified Karmali agar, increasing isolation rate and selectivity for C. jejuni and C. coli.

  17. Limitations of the clonal agar assay for the assessment of primary human ovarian tumour biopsies.

    PubMed Central

    Bertoncello, I.; Bradley, T. R.; Campbell, J. J.; Day, A. J.; McDonald, I. A.; McLeish, G. R.; Quinn, M. A.; Rome, R.; Hodgson, G. S.

    1982-01-01

    114 biopsy specimens from 70 patients with ovarian carcinoma at all stages of disease were submitted for assessment of clonogenic capacity in agar. A highly significant correlation was found between agar clonogenicity and patient survival after biopsy. However, problems related to inherent tumour heterogeneity, quality of sample and tissue disaggregation indicate that this technique may have limited applicability in the routine assessment of patients. Only 41 biopsy specimens (36%) from 31 patients (44.3%) complied with the prerequisite criteria for agar clonogenic assessment, namely: (a) the confirmed presence of malignant cells in the biopsy, (b) the ability to prepare a single-cell suspension, and (c) adequate viable cell numbers for assay. Furthermore, although the dominant patterns of agar clonogenic growth could be identified and correlated with stage of disease, the heterogeneity in both initial clonogenic capacity and "self-renewal" capacity assessed by the ability of primary clones to propagate in liquid culture and reclone in agar was too inconsistent for the assay to be used as a prognostic index for the individual patient. Images Figure PMID:7093117

  18. McKay agar enables routine quantification of the 'Streptococcus milleri' group in cystic fibrosis patients.

    PubMed

    Sibley, Christopher D; Grinwis, Margot E; Field, Tyler R; Parkins, Michael D; Norgaard, Jens C; Gregson, Daniel B; Rabin, Harvey R; Surette, Michael G

    2010-05-01

    The 'Streptococcus milleri' group (SMG) has recently been recognized as a contributor to bronchopulmonary disease in cystic fibrosis (CF). Routine detection and quantification is limited by current CF microbiology protocols. McKay agar was developed previously for the semi-selective isolation of this group. Here, McKay agar was validated against a panel of clinical SMG isolates, which revealed improved SMG recovery compared with Columbia blood agar. The effectiveness of this medium was evaluated by appending it to the standard CF sputum microbiology protocols in a clinical laboratory for a 6-month period. All unique colony types were isolated and identified by 16S rRNA gene sequencing. Whilst a wide variety of organisms were isolated, members of the SMG were the most prevalent bacteria cultured, and McKay agar allowed routine quantification of the SMG from 10(3) to >10(8) c.f.u. ml(-1) directly from sputum. All members of the SMG were detected [Streptococcus anginosus (40.7 %), Streptococcus intermedius (34.3 %) and Streptococcus constellatus (25 %)] with an overall prevalence rate of 40.6 % in our adult CF population. Without exception, samples where SMG isolates were cultured at 10(7) c.f.u. ml(-1) or greater were associated with pulmonary exacerbations. This study demonstrates that McKay agar can be used routinely to quantify the SMG from complex clinical samples.

  19. Nutrient agar with sodium chloride supplementation for presumptive detection of Moraxella catarrhalis in clinical specimens.

    PubMed

    Nishiyama, Hiroyuki; Saito, Ryoichi; Chida, Toshio; Sano, Kazumitsu; Tsuchiya, Tatsuyuki; Okamura, Noboru

    2012-04-01

    We previously reported that Nissui nutrient agar (N medium) promoted the growth of Moraxella catarrhalis but not commensal Neisseria spp. In the present study, we examined which constituent of N medium was responsible for the selective growth of M. catarrhalis using 209 M. catarrhalis and 100 commensal Neisseria spp. clinical strains. We found that peptone, but not meat extract or agar of N medium, had growth-promoting or growth-inhibiting ability with respect to M. catarrhalis and commensal Neisseria spp. Thus, we investigated the amino acid content of N peptone and found it had higher concentrations of amino acids than other commercial peptone products. On varying the sodium chloride concentration of reconstituted N medium, we noted that the concentration was an important factor in bacterial growth differences. Varying the sodium chloride concentration of other commercial nutrient agars achieved similar results to those for N medium. This is, to our knowledge, the first study observing that sodium chloride concentration is responsible for difference in growth between the two organisms. We also successfully isolated colonies of M. catarrhalis from respiratory specimens on N medium, whereas the growth of commensal Neisseria spp. was inhibited, and by adding bovine hematin and β-NAD we were able to isolate Haemophilus influenzae colonies as efficiently as with a chocolate agar. In conclusion, nutrient agar can be used as a medium for the preferential isolation of M. catarrhalis from upper respiratory tract specimens.

  20. Cell aggregation on agar as an indicator for cell-matrix adhesion: effects of opioids.

    PubMed

    Debruyne, Delphine; Mareel, Marc; Vanhoecke, Barbara; Bracke, Marc

    2009-09-01

    The slow aggregation assay is generally used to study the functionality of cell-cell adhesion complexes. Single cells are seeded on a semisolid agar substrate in a 96-well plate and the cells spontaneously aggregate. We used HEK FLAG-MOP cells that stably overexpress the mu opioid receptor and the mu-opioid-receptor-selective agonists DAMGO and morphine to study whether other factors than functionality of cell-cell adhesions complexes can contribute to changes in the pattern of slow aggregation on agar. HEK FLAG-MOP cells formed small compact aggregates. In the presence of DAMGO and morphine, larger and fewer aggregates were formed in comparison to the vehicle control. These aggregates were localized in the center of the agar surface, whereas in the vehicle control they were dispersed over the substrate. However, in suspension culture on a Gyrotory shaker, no stimulation of aggregation was observed by DAMGO and morphine, showing that opioids do not affect affinity. A dissociation experiment revealed that HEK FLAG-MOP aggregates formed in the absence or presence of opioids are resistant to de-adhesion. We demonstrated that the larger aggregates are neither the result of cell growth stimulation by DAMGO and morphine. Since manipulations of the substrate such as increasing the agar concentration or mixing agar with agarose induced the same changes in the pattern of slow aggregation as treatment with opioids, we suggest that cell-substrate adhesion may be involved in opioid-stimulated aggregation.

  1. Effects of shape and size of agar gels on heating uniformity during pulsed microwave treatment.

    PubMed

    Soto-Reyes, Nohemí; Temis-Pérez, Ana L; López-Malo, Aurelio; Rojas-Laguna, Roberto; Sosa-Morales, María Elena

    2015-05-01

    Model gel systems with different shape (sphere, cylinder, and slab) and size (180 and 290 g) were prepared with agar (5%) and sucrose (5%). Dielectric constant (ε'), loss factor (ε"), thermophysical properties, and temperature distribution of the model system were measured. Each agar model system was immersed and suspended in water, and then, heated in a microwave oven with intermittent heating until the core temperature reached 50 °C. The ε' and ε" of agar gels decreased when frequency increased. The density and thermal conductivity values of the agar gels were 1033 kg/m(3) and 0.55 W/m °C, respectively. The temperature distribution of sphere, cylinder, and slab was different when similar power doses were applied. The slab reached 50 °C in less time (10 min) and showed a more uniform heating than spheres and cylinders in both sizes. Agar model systems of 180 g heated faster than those of 290 g. The coldest point was the center of the model systems in all studied cases. Shape and size are critical food factors that affect the heating uniformity during microwave heating processes. PMID:25827444

  2. Novel grafted agar disks for the covalent immobilization of β-D-galactosidase.

    PubMed

    Wahba, Marwa I; Hassan, Mohamed E

    2015-12-01

    Novel grafted agar disks were prepared for the covalent immobilization of β-D-galactosidase (β-gal). The agar disks were activated through reacting with ethylenediamine or different molecular weights of Polyethyleneimine (PEI), followed by glutaraldehyde (GA). The modification of the agar gel and the binding of the enzyme were verified by Fourier Transform Infrared (FTIR) and elemental analysis. Moreover, the agar's activation process was optimized, and the amount of immobilized enzyme increased 3.44 folds, from 38.1 to 131.2 U/g gel, during the course of the optimization process. The immobilization of β-gal onto the activated agar disks caused its optimum temperature to increase from 45°C to 45-55°C. The optimum pH of the enzyme was also shifted towards the acidic side (3.6-4.6) after its immobilization. Additionally, the Michaelis-Menten constant (Km ) increased for the immobilized β-gal as compared to its free counterpart whereas the maximum reaction rate (Vmax ) decreased. The immobilized enzyme was also shown to retain 92.99% of its initial activity after being used for 15 consecutive times.

  3. Effects of shape and size of agar gels on heating uniformity during pulsed microwave treatment.

    PubMed

    Soto-Reyes, Nohemí; Temis-Pérez, Ana L; López-Malo, Aurelio; Rojas-Laguna, Roberto; Sosa-Morales, María Elena

    2015-05-01

    Model gel systems with different shape (sphere, cylinder, and slab) and size (180 and 290 g) were prepared with agar (5%) and sucrose (5%). Dielectric constant (ε'), loss factor (ε"), thermophysical properties, and temperature distribution of the model system were measured. Each agar model system was immersed and suspended in water, and then, heated in a microwave oven with intermittent heating until the core temperature reached 50 °C. The ε' and ε" of agar gels decreased when frequency increased. The density and thermal conductivity values of the agar gels were 1033 kg/m(3) and 0.55 W/m °C, respectively. The temperature distribution of sphere, cylinder, and slab was different when similar power doses were applied. The slab reached 50 °C in less time (10 min) and showed a more uniform heating than spheres and cylinders in both sizes. Agar model systems of 180 g heated faster than those of 290 g. The coldest point was the center of the model systems in all studied cases. Shape and size are critical food factors that affect the heating uniformity during microwave heating processes.

  4. Strategies to improve the mechanical strength and water resistance of agar films for food packaging applications.

    PubMed

    Sousa, Ana M M; Gonçalves, Maria P

    2015-11-01

    Agar films possess several properties adequate for food packaging applications. However, their high cost-production and quality variations caused by physiological and environmental factors affecting wild seaweeds make them less attractive for industries. In this work, native (NA) and alkali-modified (AA) agars obtained from sustainably grown seaweeds (integrated multi-trophic aquaculture) were mixed with locust bean gum (LBG) to make 'knife-coated' films with fixed final concentration (1 wt%) and variable agar/LBG ratios. Agar films were easier to process upon LBG addition (viscosity increase and gelling character decrease of the film-forming solutions observed by dynamic oscillatory and steady shear measurements). The mechanical properties and water resistance were optimal for films with 50 and/or 75% LBG contents and best in the case of NA (cheaper to extract). These findings can help reduce the cost-production of agar packaging films. Moreover, the controlled cultivation of seaweeds can provide continuous and reliable feedstock for transformation industries. PMID:26256341

  5. Modification of Karmali agar by supplementation with potassium clavulanate for the isolation of Campylobacter from chicken carcass rinses.

    PubMed

    Chon, Jung-Whan; Kim, Hong-Seok; Kim, Dong-Hyeon; Kim, Hyunsook; Choi, In-Soo; Oh, Deog-Hwan; Seo, Kun-Ho

    2014-07-01

    The detection ability and selectivity of Karmali agar was improved by supplementation of an extended-spectrum β-lactamase inhibitor, potassium clavulanate. The optimum concentration of potassium clavulanate (0.5 μg/ml) in Karmali agar was determined by inoculation of 50 Campylobacter and 30 extended-spectrum β-lactamase-producing E. coli strains onto normal and modified Karmali agar containing various concentrations of the agent. Eighty retail carcasses were rinsed with 400 ml of buffered peptone water. The rinse samples were enriched in 2 × blood-free Bolton enrichment broth at 42°C for 48 h and then were streaked onto normal and modified Karmali agar containing 0.5 μg/ml potassium clavulanate. The suspicious colonies were subcultured on Columbia blood agar and confirmed by colony PCR. In chicken carcass samples, the modified Karmali agar showed a significantly greater isolation rate than normal Karmali agar (42.5 versus 21.3%; P < 0.05). Furthermore, the selectivity of the modified Karmali agar was also significantly higher (P < 0.05) than that of the normal Karmali agar, as seen by comparison of the number of contaminated agar plates (83.8 versus 97.5%) and the growth index (1.67 versus 2.91) of the non-Campylobacter colonies.

  6. Comparison of effectiveness of wood decay fungi maintained by annual subculture on agar and stored in sterile water for 18 years.

    PubMed

    Richter, Dana L; Kangas, Laura C; Smith, Jill K; Laks, Peter E

    2010-03-01

    Fourteen isolates of basidiomycete decay fungi (12 species) were maintained for 18 years on agar slants transferred annually and also stored as mycelium-agar cores under cold sterile water without subculture. Isolates stored by each method were evaluated for decay effectiveness using a standard laboratory accelerated soil-block decay test. Effectiveness was measured by mean percent mass loss of wood blocks. There was no significant difference (p < or = 0.05) in decay effectiveness between storage methods for 12 of the fungus isolates tested. For the 2 fungi that showed a significant difference in the amount of decay with respect to storage method, 1 fungus (Fomitopsis lilacinogilva) produced more decay by the strain maintained as an agar slant, while the other fungus (Trametes versicolor) produced more decay by the strain stored in sterile water. Results suggested that storage under sterile water is an easy and effective method to store isolates of decay fungi for long periods, but as with any microbial storage method, careful monitoring of isolates upon revival is necessary.

  7. Impact of dilution on microbial community structure and functional potential: comparison of numerical simulations and batch culture experiments

    NASA Technical Reports Server (NTRS)

    Franklin, R. B.; Garland, J. L.; Bolster, C. H.; Mills, A. L.

    2001-01-01

    A series of microcosm experiments was performed using serial dilutions of a sewage microbial community to inoculate a set of batch cultures in sterile sewage. After inoculation, the dilution-defined communities were allowed to regrow for several days and a number of community attributes were measured in the regrown assemblages. Based upon a set of numerical simulations, community structure was expected to differ along the dilution gradient; the greatest differences in structure were anticipated between the undiluted-low-dilution communities and the communities regrown from the very dilute (more than 10(-4)) inocula. Furthermore, some differences were expected among the lower-dilution treatments (e.g., between undiluted and 10(-1)) depending upon the evenness of the original community. In general, each of the procedures used to examine the experimental community structures separated the communities into at least two, often three, distinct groups. The groupings were consistent with the simulated dilution of a mixture of organisms with a very uneven distribution. Significant differences in community structure were detected with genetic (amplified fragment length polymorphism and terminal restriction fragment length polymorphism), physiological (community level physiological profiling), and culture-based (colony morphology on R2A agar) measurements. Along with differences in community structure, differences in community size (acridine orange direct counting), composition (ratio of sewage medium counts to R2A counts, monitoring of each colony morphology across the treatments), and metabolic redundancy (i.e., generalist versus specialist) were also observed, suggesting that the differences in structure and diversity of communities maintained in the same environment can be manifested as differences in community organization and function.

  8. Digital image quantification of siderophores on agar plates.

    PubMed

    Andrews, Megan Y; Santelli, Cara M; Duckworth, Owen W

    2016-03-01

    This article presents visual image data and detailed methodology for the use of a new method for quantifying the exudation of siderophores during fungal growth. The data include images showing time series for calibration, fungal exudation, and negative controls, as well as replication accuracy information. In addition, we provide detailed protocols for making CAS assay layer plates, the digital analysis protocol for determining area of color change, and discuss growth media that do and do not work with the layer plate method. The results of these data, their interpretation, and further discussion can be found in Andrews et al., 2016 [1]. PMID:26937467

  9. Evaluation of the antibacterial effects of vancomycin hydrochloride released from agar-gelatin-bioactive glass composites.

    PubMed

    Rivadeneira, Josefina; Di Virgilio, Ana Laura; Audisio, M Carina; Boccaccini, Aldo R; Gorustovich, Alejandro A

    2015-01-13

    The aim of this work was to evaluate the perfomance of agar-gelatin (AG) composites and AG-containing 45S5 bioactive glass (BG) microparticles (AGBG) in relation to their water uptake capacity, sustained release of a drug over time, and antibacterial effects. The composites were fabricated by the gel-casting method. To impart the local drug release capacity, vancomycin hydrochloride (VC) was loaded in the composites in concentrations of 0.5 and 1 mg ml(-1). VC release was assessed in distilled water at 37 °C up to 72 h and quantified spectrophotometrically. The antibacterial activity of composites was evaluated by the inhibition zone test and the plate count method. The experiments were performed in vitro up to 48 h on three staphylococcus strains: Staphylococcus aureus ATCC29213, S. aureus ATCC6538 and Staphylococcus epidermidis ATCC12228. The results showed that the addition of BG to AG composites did not affect the degree of water uptake. The release of VC was significantly affected by the presence of BG. VC release was higher from AGBGVC films than from AGVC ones over prolonged incubation times. Bacterial inhibition zones were found around the composites. The halos were larger when the cells were put in contact with AGVC composites than when they were put in contact with AGBGVC ones. Nevertheless, the viable count method demonstrated that the composites inhibited Staphylococcus cell growth with no statistical differences. In conclusion, the addition of BG did not reflect an improvement in the parameters studied. On the other hand, composites loaded with VC would have a role in prophylaxis against bacterial infection.

  10. Effect of Diethylaminoethyl Dextran on the Growth of Mycoplasma in Agar

    PubMed Central

    Tauraso, Nicola M.

    1967-01-01

    The growth of certain strains of Mycoplasma is inhibited by substances present in commercial agar preparations. The addition of diethylaminoethyl (DEAE) dextran (10 mg per 100 ml) to agar media appears to enhance the growth of some strains. Of eight strains initially tested, the presence of DEAE dextran grossly enhanced the growth of three strains. One strain appeared not to be affected, and a clearly enhancing effect was not evident with four strains. Quantitative studies revealed that growth enhancement varied from 10 colony-forming units (CFU) for M. hominis type II (strain Campo) to 103.3 CFU for M. pulmonis (strain 880). The growth-enhancing effect is probably due to the ability of DEAE dextran to bind the sulfated polysaccharide moieties in agar and not to the DEAE dextran, per se. Images PMID:6025444

  11. Expression of an accessory cell phenotype by hairy cells during lymphocyte colony formation in agar culture.

    PubMed

    Farcet, J P; Gourdin, M F; Testa, U; Andre, C; Jouault, H; Reyes, F

    1983-01-01

    Human T lymphocytes require the cooperation of accessory cells to generate lymphocyte colonies in agar culture under PHA stimulation. Various hairy cell enriched fractions, as well as normal monocytes, have been found to be able to initiate colony formation by normal lymphocytes. Leukemic monocytes from CMML patients were also effective, but not the leukemic lymphocytes from CLL patients. The phenotype expressed by HC in agar colonies was further studied using cell surface and enzymatic markers. We have concluded that HC in agar culture in the presence of both normal T lymphocytes and PHA lose the B phenotype that they express in vivo and function like an accessory cell in contrast to normal or leukemic B lymphocytes. PMID:6601222

  12. A helium-3/helium-4 dilution cryocooler for operation in zero gravity

    NASA Technical Reports Server (NTRS)

    Hendricks, John B.

    1988-01-01

    This research effort covered the development of He-3/He-4 dilution cryocooler cycles for use in zero gravity. The dilution cryocooler is currently the method of choice for producing temperatures below 0.3 Kelvin in the laboratory. However, the current dilution cryocooler depends on gravity for their operation, so some modification is required for zero gravity operation. In this effort, we have demonstrated, by analysis, that the zero gravity dilution cryocooler is feasible. We have developed a cycle that uses He-3 circulation, and an alternate cycle that uses superfluid He-4 circulation. The key elements of both cycles were demonstrated experimentally. The development of a true 'zero-gravity' dilution cryocooler is now possible, and should be undertaken in a follow-on effort.

  13. Radiation effects on agar, alginates and carrageenan to be used as food additives

    NASA Astrophysics Data System (ADS)

    Aliste, A. J. A. J.; Vieira, F. F. F. F.; Del Mastro, N. L. N. L.

    2000-03-01

    Agar, alginates and carrageenan are hydrocolloids that induce stabilization of physical properties of the food product during shelf life and prevention of undesirable changes such as moisture migration, gas cell coalescence or textural profile changes. In this work, agar, alginates and carrageenan was irradiated as powder with different doses (0-10 kGy) of Co-60 and the rheological functional performance of water solutions of these irradiated additives was studied. The results are analyzed taking in account the future applications of those additives in irradiated foods.

  14. Agar-polydimethylsiloxane devices for quantitative investigation of oviposition behaviour of adult Drosophila melanogaster

    PubMed Central

    Leung, Jacob C. K.; Taylor-Kamall, Rhodri W.; Hilliker, Arthur J.; Rezai, Pouya

    2015-01-01

    Drosophila melanogaster (fruit fly) is a model organism and its behaviours including oviposition (egg-laying) on agar substrates have been widely used for assessment of a variety of biological processes in flies. Physical and chemical properties of the substrate are the dominant factors affecting Drosophila's oviposition, but they have not been investigated precisely and parametrically with the existing manual approaches. As a result, many behavioral questions about Drosophila oviposition, such as the combined effects of the aforementioned substrate properties (e.g., exposure area, sugar content, and stiffness) on oviposition and viability, and their threshold values, are yet to be answered. In this paper, we have devised a simple, easily implementable, and novel methodology that allows for modification of physical and chemical composition of agar substrates in order to quantitatively study survival and oviposition of adult fruit flies in an accurate and repeatable manner. Agar substrates have been modified by surface patterning using single and hexagonally arrayed through-hole polydimethylsiloxane (PDMS) membranes with various diameters and interspacing, as well as by substrate stiffness and sugar content modification via alteration of chemical components. While pure PDMS substrates showed a significant lethal effect on flies, a 0.5 mm diameter through-hole access to agar was found to abruptly increase the survival of adult flies to more than 93%. Flies avoided ovipositing on pure PDMS and on top of substrates with 0.5 mm diameter agar exposure areas. At a hole diameter of 2 mm (i.e., 0.25% exposure area) or larger, eggs were observed to be laid predominately inside the through-holes and along the edges of the PDMS-agar interface, showing a trending increase in site selection with 4 mm (i.e., 1% exposure area threshold) demonstrating natural oviposition rates similar to pure agar. The surface-modified agar-PDMS hybrid devices and the threshold values

  15. Serial dilution via surface energy trap-assisted magnetic droplet manipulation.

    PubMed

    Zhang, Yi; Shin, Dong Jin; Wang, Tza-Huei

    2013-12-21

    This paper demonstrates a facile method of generating precise serial dilutions in the form of droplets on an open surface platform. The method relies on the use of surface energy traps (SETs), etched areas of high surface energy on a Teflon coated glass substrate, to assist in the magnetic manipulation of droplets to meter and dispense liquid of defined volumes for the preparation of serial dilutions. The volume of the dispensed liquid can be precisely controlled by the size of the SETs, facilitating generation of concentration profiles of high linearity. We have applied this approach to the generation of serial dilutions of antibiotics for anti-microbial susceptibility testing (AST).

  16. Revisiting the Dilution Procedure Used To Manipulate Microbial Biodiversity in Terrestrial Systems

    PubMed Central

    Yan, Yan; Kuramae, Eiko E.; Klinkhamer, Peter G. L.

    2015-01-01

    It is hard to assess experimentally the importance of microbial diversity in soil for the functioning of terrestrial ecosystems. An approach that is often used to make such assessment is the so-called dilution method. This method is based on the assumption that the biodiversity of the microbial community is reduced after dilution of a soil suspension and that the reduced diversity persists after incubation of more or less diluted inocula in soil. However, little is known about how the communities develop in soil after inoculation. In this study, serial dilutions of a soil suspension were made and reinoculated into the original soil previously sterilized by gamma irradiation. We determined the structure of the microbial communities in the suspensions and in the inoculated soils using 454-pyrosequencing of 16S rRNA genes. Upon dilution, several diversity indices showed that, indeed, the diversity of the bacterial communities in the suspensions decreased dramatically, with Proteobacteria as the dominant phylum of bacteria detected in all dilutions. The structure of the microbial community was changed considerably in soil, with Proteobacteria, Bacteroidetes, and Verrucomicrobia as the dominant groups in most diluted samples, indicating the importance of soil-related mechanisms operating in the assembly of the communities. We found unique operational taxonomic units (OTUs) even in the highest dilution in both the suspensions and the incubated soil samples. We conclude that the dilution approach reduces the diversity of microbial communities in soil samples but that it does not allow accurate predictions of the community assemblage during incubation of (diluted) suspensions in soil. PMID:25888175

  17. Revisiting the dilution procedure used to manipulate microbial biodiversity in terrestrial systems.

    PubMed

    Yan, Yan; Kuramae, Eiko E; Klinkhamer, Peter G L; van Veen, Johannes A

    2015-07-01

    It is hard to assess experimentally the importance of microbial diversity in soil for the functioning of terrestrial ecosystems. An approach that is often used to make such assessment is the so-called dilution method. This method is based on the assumption that the biodiversity of the microbial community is reduced after dilution of a soil suspension and that the reduced diversity persists after incubation of more or less diluted inocula in soil. However, little is known about how the communities develop in soil after inoculation. In this study, serial dilutions of a soil suspension were made and reinoculated into the original soil previously sterilized by gamma irradiation. We determined the structure of the microbial communities in the suspensions and in the inoculated soils using 454-pyrosequencing of 16S rRNA genes. Upon dilution, several diversity indices showed that, indeed, the diversity of the bacterial communities in the suspensions decreased dramatically, with Proteobacteria as the dominant phylum of bacteria detected in all dilutions. The structure of the microbial community was changed considerably in soil, with Proteobacteria, Bacteroidetes, and Verrucomicrobia as the dominant groups in most diluted samples, indicating the importance of soil-related mechanisms operating in the assembly of the communities. We found unique operational taxonomic units (OTUs) even in the highest dilution in both the suspensions and the incubated soil samples. We conclude that the dilution approach reduces the diversity of microbial communities in soil samples but that it does not allow accurate predictions of the community assemblage during incubation of (diluted) suspensions in soil.

  18. Quantitative Analysis by Isotopic Dilution Using Mass Spectroscopy: The Determination of Caffeine by GC-MS.

    ERIC Educational Resources Information Center

    Hill, Devon W.; And Others

    1988-01-01

    Describes a laboratory technique for quantitative analysis of caffeine by an isotopic dilution method for coupled gas chromatography-mass spectroscopy. Discusses caffeine analysis and experimental methodology. Lists sample caffeine concentrations found in common products. (MVL)

  19. Declining ecosystem health and the dilution effect.

    PubMed

    Khalil, Hussein; Ecke, Frauke; Evander, Magnus; Magnusson, Magnus; Hörnfeldt, Birger

    2016-01-01

    The "dilution effect" implies that where species vary in susceptibility to infection by a pathogen, higher diversity often leads to lower infection prevalence in hosts. For directly transmitted pathogens, non-host species may "dilute" infection directly (1) and indirectly (2). Competitors and predators may (1) alter host behavior to reduce pathogen transmission or (2) reduce host density. In a well-studied system, we tested the dilution of the zoonotic Puumala hantavirus (PUUV) in bank voles (Myodes glareolus) by two competitors and a predator. Our study was based on long-term PUUV infection data (2003-2013) in northern Sweden. The field vole (Microtus agrestis) and the common shrew (Sorex araneus) are bank vole competitors and Tengmalm's owl (Aegolius funereus) is a main predator of bank voles. Infection probability in bank voles decreased when common shrew density increased, suggesting that common shrews reduced PUUV transmission. Field voles suppressed bank vole density in meadows and clear-cuts and indirectly diluted PUUV infection. Further, Tengmalm's owl decline in 1980-2013 may have contributed to higher PUUV infection rates in bank voles in 2003-2013 compared to 1979-1986. Our study provides further evidence for dilution effect and suggests that owls may have an important role in reducing disease risk. PMID:27499001

  20. Time- and media-saving testing and identification of microorganisms by multipoint inoculation on undivided agar plates.

    PubMed Central

    Burman, L G; Ostensson, R

    1978-01-01

    Motility and various biochemical activities of isolates of bacteria and yeasts were tested on undivided agar plates by using a simple, manually operated multipoint inoculation apparatus that allowed the analysis of 25 isolates per 9-cm-diameter petri plate. Fermentation of all 17 carbohydrates tested as well as 13 other biochemical activities commonly used for identification of bacteria were readily demonstrated by the multipoint inoculation plate method, and the results agreed very well with those of conventional tube tests. In addition to speedy inoculation and low cost of materials, the multipoint inoculation plate method offers several other advantages when compared with conventional tube tests or with some of the manufactured test kits currently available for recognizing members of the family Enterobacteriaceae. Images PMID:359588