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Sample records for agar diffusion bioassay

  1. 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. PMID:26405525

  2. 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

  3. Disk Agar Diffusion Susceptibility Testing of Yeasts

    PubMed Central

    Saubolle, Michael A.; Hoeprich, Paul D.

    1978-01-01

    A disk agar diffusion method was developed for testing the susceptibility of rapidly growing yeasts in vitro. A totally defined, completely synthetic agar culture medium (synthetic amino acid medium, fungal) and clinical isolates of Candida spp. and Torulopsis glabrata were used. Turbidimetric adjustment of cell suspensions resulted in standard, reproducible inocula, which gave sharp, clear zones of inhibition when applied by an agar overlay method. Optimal disk loads were determined for amphotericin B, amphotericin B methyl ester, 5-fluorocytosine, clotrimazole, and miconazole. Disk potencies were stable over a 2-month period when stored in a vacuum desiccator at −30°C. Using an error ratebounded classification, the zones of inhibition were correlated with both broth dilution and agar dilution minimum inhibitory concentrations (MICs). With amphotericin B and amphotericin B methyl ester, all isolates were susceptible, yielding zone diameters which clustered within 5 mm. Overall correlations between zone diameters and broth dilution MICs with 5-fluorocytosine, miconazole, and clotrimazole were 97, 96, and 82% (excluding T. glabrata), respectively; correlations of zone diameters with agar dilution MICs were 96, 92, and 88%, respectively. Disk diffusion susceptibility testing of yeasts appears to be generally applicable. However, when results are equivocal, quantitative test methods should be used. PMID:568910

  4. Improved agar diffusion method for detecting residual antimicrobial agents.

    PubMed

    Tsai, C E; Kondo, F

    2001-03-01

    The improved agar diffusion method for determination of residual antimicrobial agents was investigated, and the sensitivities of various combinations of test organisms and assay media were determined using 7 organisms, 5 media, and 31 antimicrobial agents. Bacillus stearothermophilus and synthetic assay medium (SAM) showed the greatest sensitivity for screening penicillins (penicillin G and ampicillin). The combination of Bacillus subtilis and minimum medium (MM) was the most sensitive for tetracyclines (oxytetracycline and chlortetracycline), B. stearothermophilus and SAM or Micrococcus luteus and Mueller-Hinton agar (MHA) for detecting tylosin and erythromycin, B. subtilis and MHA for aminoglycosides (streptomycin, kanamycin, gentamicin, and dihydrostreptomycin), B. stearothermophilus and SAM for polyethers (salinomycin and lasalocid), and B. subtilis and MM or Clostridium perfringens and GAM for polypeptides (thiopeptin, enramycin, virginiamycin, and bacitracin). However, gram-negative bacterium Escherichia coli ATCC 27166 and MM were better for screening for colistin and polymixin-B. For detecting the synthetic drugs tested, the best combination was B. subtilis and MM for sulfonamides, E. coli 27166 and MM for quinolones (oxolinic acid and nalidixic acid), B. subtilis and MM for furans (furazolidone), and the bioluminescent bacterium Photobacterium phosphoreum and luminescence assay medium for chloramphenicol and oxolinic acid. The results showed that the use of four assay plates, B. stearothermophilus and SAM, B. subtilis and MM, M. luteus and MHA, and E. coli 27166 and MM, was superior to the currently available techniques for screening for residual antimicrobial agents in edible animal tissues. PMID:11252480

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

    PubMed

    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 (R (2) in the linear experiment of Na2S2O3 was 0.994; R (2) 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

  6. 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

  7. 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...

  8. Diffusion of Methylene Blue in Phantoms of Agar Using a Photoacoustic Technique

    NASA Astrophysics Data System (ADS)

    Vilca-Quispe, L.; Alvarado-Gil, J. J.; Quintana, P.; Ordonez-Miranda, J.

    2010-05-01

    In this work, the kinetics of diffusion of methylene blue in agar aqueous solution is studied using a photoacoustic technique. Two agar phantoms solutions in water with a relation of mass/volume of 0.01% and 0.05% were analyzed. The study was performed using a modified Rosencwaig photoacoustic cell that is enclosed by transparent windows, on both sides. The sample is deposited directly on top of the upper window. A red light beam, at a fixed modulation frequency, is sent through the lower window illuminating the sample and inducing the photoacoustic effect inside the closed chamber of the cell. At the beginning of the experiment, a droplet of 100μL of agar solution is deposited; afterwards, the signal stabilizes, and 10μL of methylene blue aqueous solution (0.0125 g · mL-1) is added to the surface of the agar. During the first seconds of the experiment, the photoacoustic signal amplitude increases followed by a gradual and long decay. Results for modulation frequencies in the range from 10Hz to 80Hz for both agar concentrations are presented. A simple theoretical approach is presented to analyze the experimental data. It is demonstrated that the kinetics of the process can be parameterized as a function of the changes of an effective optical absorption coefficient. From these results, the characteristic time, in which the dye diffusion process stabilizes, is obtained. It is found that this time is larger for samples with a higher agar concentration. These differences provide important results for biomedical sciences in which agar gels are used as phantoms resembling some of the properties of living organs and tissues.

  9. An agar diffusion study comparing the antimicrobial activity of Nanoseal with some other endodontic sealers.

    PubMed

    Aal-Saraj, Ali Burak; Ariffin, Zaihan; Masudi, Sam'an Malik

    2012-08-01

    The aim of this study was to evaluate the antimicrobial activity of a new experimental nano-hydroxyapatite epoxy resin-based sealer (Nanoseal) with several other commercially available sealers; AH26, Tubliseal, Sealapex and Roekoseal against Enterococcus faecalis, Pseudomonas aeruginosa, Streptococcus mutans, Streptococcus sobrinus and Escherichia coli for up to 7 days. Agar diffusion was used in this study. Fifty Muller-Hinton agar plates were prepared and divided into five experimental groups (n = 10), for each micro-organism. Another 10 agar plates were used as positive and negative controls. Endodontic sealers were tested against each micro-organism. Inhibition zones produced were recorded. The results of this study showed that all test materials exhibited inhibition zones towards the tested micro-organisms for 7 days except for Roekoseal, which showed no inhibition zones. Nanoseal and AH26 exhibited similar zones of inhibition. Significant difference was found between Nanoseal and the other tested sealers (P < 0.001). PMID:22827817

  10. Cytotoxicity of ferrite particles by MTT and agar diffusion methods for hyperthermic application

    NASA Astrophysics Data System (ADS)

    Kim, Dong-Hyun; Lee, Se-Ho; Kim, Kyoung-Nam; Kim, Kwang-Mahn; Shim, In-Bo; Lee, Yong-Keun

    2005-05-01

    We investigated the cytotoxicity of the prepared various ferrites (Fe-, Li-, Ni/Zn/Cu-, Ba-, Sr-, Co-, Co/Ni-ferrites) using MTT assay as well as agar diffusion method. Their cytotoxicity was compared with that of alginate-encapsulated ferrites. In the MTT assay, Fe 3O 4 and SrFe 12O 19 ferrite showed the highest cell viability of 90%. Alginate-encapsulated Ba-ferrite was ranked mildly cytotoxic, whereas their ferrite particles were ranked cytotoxic.

  11. 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

  12. Serotyping reanalysis of unserotypable Actinobacillus pleuropneumoniae isolates by agar gel diffusion test.

    PubMed

    Morioka, Ayako; Shimazaki, Yoko; Uchiyama, Mariko; Suzuki, Shoko

    2016-05-01

    We observed increasing unserotypable (UT) Actinobacillus pleuropneumoniae isolates using agar gel diffusion (AGD) test. To reanalyze their serovar, we performed rapid slide agglutination (RSA) test and multiplex PCR for 47 UT isolates. Of these, 25 were serovar 1 (UT-serovar 1), 20 were serovar 2 (UT-serovar 2) and 2 were serovar 15 (UT-serovar 15). We examined serotyping antigen extraction temperature to determine heat influence. UT-serovar 1 and 15 were influenced by heat, because their precipitation lines were observed in the case of low antigen extraction temperature. To investigate the relationship between antigenicity and genotype, we performed pulsed-field gel electrophoresis (PFGE) analysis using UT-serovar 2 and 15. The predominant PFGE pattern of UT-serovar 2 was identical to that of serovar 2. PMID:26726101

  13. Serotyping reanalysis of unserotypable Actinobacillus pleuropneumoniae isolates by agar gel diffusion test

    PubMed Central

    MORIOKA, Ayako; SHIMAZAKI, Yoko; UCHIYAMA, Mariko; SUZUKI, Shoko

    2016-01-01

    We observed increasing unserotypable (UT) Actinobacillus pleuropneumoniae isolates using agar gel diffusion (AGD) test. To reanalyze their serovar, we performed rapid slide agglutination (RSA) test and multiplex PCR for 47 UT isolates. Of these, 25 were serovar 1 (UT-serovar 1), 20 were serovar 2 (UT-serovar 2) and 2 were serovar 15 (UT-serovar 15). We examined serotyping antigen extraction temperature to determine heat influence. UT-serovar 1 and 15 were influenced by heat, because their precipitation lines were observed in the case of low antigen extraction temperature. To investigate the relationship between antigenicity and genotype, we performed pulsed-field gel electrophoresis (PFGE) analysis using UT-serovar 2 and 15. The predominant PFGE pattern of UT-serovar 2 was identical to that of serovar 2. PMID:26726101

  14. 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

  15. 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.

  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. Inaccuracy of the Disk Diffusion Method Compared with the Agar Dilution Method for Susceptibility Testing of Campylobacter spp.

    PubMed Central

    Kotilainen, Pirkko; Puukka, Pauli; Nakari, Ulla-Maija; Siitonen, Anja; Eerola, Erkki; Huovinen, Pentti; Hakanen, Antti J.

    2012-01-01

    The agar dilution method has been standardized by the CLSI for the susceptibility testing of Campylobacter species, and according to these standards, the disk diffusion method should be used only in screening for macrolide and ciprofloxacin resistance. Nevertheless, the disk diffusion test is currently widely used, since it is easy to perform in clinical microbiology laboratories. In this study, the disk diffusion method was compared to the agar dilution method by analyzing the in vitro activities of seven antimicrobial agents against 174 Campylobacter strains collected in Finland between 2003 and 2008. Recommendations of the CLSI were followed using Mueller-Hinton agar plates with 5% of sheep blood. For each strain, the disk diffusion tests were performed two to four times. Of the 33 erythromycin-resistant strains (MIC, ≥16 μg/ml), 24 (73%) constantly showed a 6-mm erythromycin inhibition zone (i.e., no inhibition), while for seven strains the inhibition zone varied from 6 to 44 mm in repeated measurements. Among the 141 erythromycin-susceptible strains (MIC, <16 μg/ml), erythromycin inhibition zones varied between 6 and 61 mm. Of the 87 ciprofloxacin-resistant strains, 47 (54%) showed 6-mm inhibition zones, while 40 strains showed inhibition zones between 6 and 60 mm. Significant differences between the repetitions were observed in the disk diffusion for all antimicrobial agents and all strains except for the macrolide-resistant strains regarding the macrolides. For 17 (10%) strains, the variation in repeated measurements was substantial. These results show that the disk diffusion method may not be a reliable tool for the susceptibility testing of Campylobacter spp. Further studies are needed to assess whether the disk diffusion test could be improved or whether all susceptibilities of campylobacters should be tested using an MIC-based method. PMID:22075583

  18. Identification of Actinomyces israelii and Actinomyces naeslundii by Fluorescent-Antibody and Agar-Gel Diffusion Techniques1

    PubMed Central

    Lambert, Frank W.; Brown, June M.; Georg, Lucille K.

    1967-01-01

    This study was an attempt to develop a fluorescent-antibody (FA) test to differentiate Actinomyces israelii and A. naeslundii as an aid in their laboratory identification. Two strains of A. israelii (X522 and A601) and two strains of A. naeslundii (X454 and X600), which had received intensive study by several investigators, were used for the immunization of rabbits. Working titers, based on tests with antigens prepared from the homologous strains and from well-established heterologous strains, were determined for each labeled antibody preparation. These conjugates and their normal serum control conjugates were used separately to stain 85 cultures of Actinomyes species and 23 strains of other species that might be confused with them. Acetone-precipitated soluble antigens from these same strains were tested with different antisera in the agar-gel diffusion test. Results showed that A. israelii (X522 and A601) and A. naeslundii (X454 and X600) labeled antiglobulins, when used at their working titers, stained most strains of their homologous species. Agar-gel diffusion results showed general agreement with those of the FA tests. The two tests appear to be equal in sensitivity, but the FA test is more specific, since several cross-reactions were noted with the agar-gel diffusion test whereas no cross-reactions were obtained with the FA reagents. Agar-gel and FA studies suggest that at least two serotypes of A. israelii may be associated with human disease. Although the majority of strains tested in this study appear to belong to a common serotype, “serotype 1,” two strains of an apparent second serotype, “serotype 2,” were encountered. FA staining of tissue impression smears from experimentally infected mice was successful when a counterstain, Evans Blue dye, was used. PMID:4964473

  19. Use of agar diffusion assay to evaluate bactericidal activity of formulations of alkaline salts of fatty acids against bacteria associated with poultry processing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The agar diffusion assay was used to examine antibacterial activity of alkaline salts of fatty acids (FA). Wells in agar media seeded with bacteria were filled with FA-potassium hydroxide (KOH) solutions, plates were incubated, and zones of inhibition were measured. The relationship between bacteric...

  20. 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

  1. Use of agar diffusion assay to measure bactericidal activity of alkaline salts of fatty acids against bacteria associated with poultry processing

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The agar diffusion assay was used to examine antibacterial activity of alkaline salts of caproic, caprylic, capric, lauric, and myristic acids. A 0.5M concentration of each fatty acid was dissolved in 1.0 M potassium hydroxide (KOH), and pH of the mixtures was adjusted to 10.5 with citric acid. Solu...

  2. 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

  3. Comparison of different agar diffusion methods for the detection of residues in the kidneys of pigs treated with antimicrobial drugs.

    PubMed

    Korkeala, H; Sorvettula, O; Mäki-Petäys, O; Hirn, J

    1983-01-01

    Residue analyses of the kidneys of twenty-six pigs treated with various antimicrobial drugs 20 h before slaughter and of eleven untreated pigs were performed. The effects of storage temperature of the kidneys, and of sampling location, on the residue analysis were also studied. No method alone was sufficient for the detection of residues. Oxytetracycline residues could be detected at pH 6, dihydrostreptomycin residues at pH 8, and sulphonamide residues if trimethoprim was present in the medium. Chloramphenicol, penicillin G procaine, tylosin and lincomycin residues were not detectable with the methods used. The concentration of ampicillin decreased during the storage of samples at +4°C. Most methods also yielded zones of inhibition for the frozen kidneys from untreated pigs. It seems necessary to use agar media of two different pH values: the addition of trimethoprim to the medium is also needed. The use of fresh pig kidneys, and samples containing both kidney medulla and kidney cortex, is recommended in residue analysis. PMID:22055926

  4. 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...

  5. 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...

  6. Kinetic microplate bioassays for relative potency of antibiotics improved by partial Least Square (PLS) regression.

    PubMed

    Francisco, Fabiane Lacerda; Saviano, Alessandro Morais; Almeida, Túlia de Souza Botelho; Lourenço, Felipe Rebello

    2016-05-01

    Microbiological assays are widely used to estimate the relative potencies of antibiotics in order to guarantee the efficacy, safety, and quality of drug products. Despite of the advantages of turbidimetric bioassays when compared to other methods, it has limitations concerning the linearity and range of the dose-response curve determination. Here, we proposed to use partial least squares (PLS) regression to solve these limitations and to improve the prediction of relative potencies of antibiotics. Kinetic-reading microplate turbidimetric bioassays for apramacyin and vancomycin were performed using Escherichia coli (ATCC 8739) and Bacillus subtilis (ATCC 6633), respectively. Microbial growths were measured as absorbance up to 180 and 300min for apramycin and vancomycin turbidimetric bioassays, respectively. Conventional dose-response curves (absorbances or area under the microbial growth curve vs. log of antibiotic concentration) showed significant regression, however there were significant deviation of linearity. Thus, they could not be used for relative potency estimations. PLS regression allowed us to construct a predictive model for estimating the relative potencies of apramycin and vancomycin without over-fitting and it improved the linear range of turbidimetric bioassay. In addition, PLS regression provided predictions of relative potencies equivalent to those obtained from agar diffusion official methods. Therefore, we conclude that PLS regression may be used to estimate the relative potencies of antibiotics with significant advantages when compared to conventional dose-response curve determination. PMID:26971814

  7. Electro-osmosis in gels: Application to Agar-Agar

    NASA Astrophysics Data System (ADS)

    Cherblanc, Fabien; Boscus, Jérôme; Bénet, Jean-Claude

    2008-10-01

    Widely used in food- and bio-engineering as a reference material, Agar-Agar gel is the focus of an experimental investigation concerning the electro-osmosis phenomenon. After presenting the experimental methods, one trial is discussed in detail. A fair reproducibility of results is obtained, and the averaged electro-osmotic permeability is provided. This value lies in the range generally measured on various kind of soils, even if Agar-Agar gel does not share any micro-structural characteristics with soils. To cite this article: F. Cherblanc et al., C. R. Mecanique 336 (2008).

  8. 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...

  9. 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 CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS §...

  10. 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 CONSUMPTION (CONTINUED) DIRECT FOOD SUBSTANCES AFFIRMED AS GENERALLY RECOGNIZED AS SAFE Listing of Specific Substances Affirmed as GRAS § 184.1115...

  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. Automatic agar tray inoculation device

    NASA Technical Reports Server (NTRS)

    Wilkins, J. R.; Mills, S. M.

    1972-01-01

    Automatic agar tray inoculation device is simple in design and foolproof in operation. It employs either conventional inoculating loop or cotton swab for uniform inoculation of agar media, and it allows technician to carry on with other activities while tray is being inoculated.

  14. Fastidious anaerobe agar compared with Wilkins-Chalgren agar, brain heart infusion agar, and brucella agar for susceptibility testing of Fusobacterium species.

    PubMed

    Brazier, J S; Goldstein, E J; Citron, D M; Ostovari, M I

    1990-11-01

    Fastidious anaerobe agar supported the growth of 82 strains of fusobacteria better than brain heart infusion agar, brucella agar, and Wilkins-Chalgren agar. Fastidious anaerobe agar showed less hazing and fewer tailing endpoints with beta-lactam antibiotics. Whole-blood supplementation improved the performance of all media. Wilkins-Chalgren agar without blood failed to support the growth of 17% of the strains. All Fusobacterium ulcerans strains were resistant to clindamycin. PMID:2073122

  15. 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)...

  16. 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)...

  17. Thermal characterization of magnetically aligned carbonyl iron/agar composites.

    PubMed

    Diaz-Bleis, D; Vales-Pinzón, C; Freile-Pelegrín, Y; Alvarado-Gil, J J

    2014-01-01

    Composites of magnetic particles into polymeric matrices have received increasing research interest due to their capacity to respond to external magnetic or electromagnetic fields. In this study, agar from Gelidium robustum has been chosen as natural biocompatible polymer to build the matrix of the magnetic carbonyl iron particles (CIP) for their uses in biomedical fields. Heat transfer behavior of the CIP-agar composites containing different concentrations (5, 10, 15, 20, 25 and 30% w/w) of magnetically aligned and non-aligned CIP in the agar matrix was studied using photothermal radiometry (PTR) in the back-propagation emission configuration. The morphology of the CIP-agar composites with aligned and non-aligned CIP under magnetic field was also evaluated by scanning electron microscopy (SEM). The results revealed a dominant effect of CIP concentration over the alignment patterns induced by the magnetic field, which agrees with the behavior of the thermal diffusivity and thermal conductivity. Agar served as a perfect matrix to be used with CIP, and CIP-agar composites magnetically aligned at 20% CIP concentration can be considered as promising 'smart' material for hyperthermia treatments in the biomedical field. PMID:24274482

  18. 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

  19. Nanovehicles based Bioassay Labels

    SciTech Connect

    Liu, Guodong; Wang, Jun; Wu, Hong; Lin, Ying-Ying; Lin, Yuehe

    2007-04-01

    In this article, we review recent advances of our group in nanoparticle labels based bioassay. Apoferritin and silica nanoparticles have been used as nanovehicles to load large amount of markers for highly sensitive bioassay. Markers loaded apoferritin, apoferritin-templated metallic phosphate nanoparticles, and poly [guanine] coated silica nanoparticles have been prepared, characterized and used as labels for highly sensitive bioassay of protein and DNA. Dissociation and reconstitution characteristics at different pH as well as the special cavity structure of apoferritin nanovehicle provides a simple and convenient route to prepare versatile nanoparticle labels and avoid the complicated and tedious synthesis process of conventional nanoparticle labels. The optical and electrochemical characteristics of the prepared nanoparticle labels are easily controlled by loading different optical or electrochemical markers. Additionally, the use of apoferritin nanovehicle as template for synthesis of metallic phosphate nanoparticle labels offers fast route to prepare uniform-size metallic nanoparticle labels for electrochemical bioassay and avoids the traditional harsh dissolution conditions to dissolve metallic nanoparticle tags (that is, the strong-acid dissolution of quantum dots and gold nanoparticles) during the stripping analysis step. Silica nanoparticle has also been used as nanovehicle to carry thousands of poly [guanine] tracers, which was used to enhance the oxidation current of Ru(bpy)32+, resulting in enhanced sensitivity of electrochemical immunoassay. The new nanovehicle-based labels have been used for highly sensitive electrochemical detection of DNA and protein biomarkers, such as tumor necrosis factor-alpha (TNF-a). The high sensitivity and selectivity make these labels a useful addition to the armory of nanoparticle-based bioassay. The new nanovehicles based labels hold great promise for multiplex protein and DNA detection and for enhancing the sensitivity

  20. 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.

  1. A RAPID, QUANTITATIVE BIOASSAY FOR DETECTING PHYTOTOXIC GASES USING STRESS-ETHYLENE

    EPA Science Inventory

    A simple bioassay for detecting phytotoxic air pollutants has been developed. Wheat (Triticum aestivum L.) seedlings growing on agar medium in test-tubes are exposed to pollutants for 2 h. Stress-ethylene produced by the seedlings during exposure to the pollutants is collected in...

  2. DEVELOPMENT OF A TEST-TUBE STRESS-ETHYLENE BIOASSAY FOR DETECTING PHYTOTOXIC GASES

    EPA Science Inventory

    A rapid, quantitative bioassay for detecting phytotoxic air pollutants has been developed. The technique uses wheat Triticum aestivum L. or tomato Lycopersicon esculentum L., seedlings growing on an agar medium in test-tubes. The seedlings are exposed to a pollutant in the test-t...

  3. Adsorptive removal of methylene blue by agar: effects of NaCl and ethanol

    PubMed Central

    2012-01-01

    Adsorption of methylene blue (MB) on agar was investigated as a function of temperature (308-328 K), different concentrations of NaCl and HCl and various weight percentages of binary mixtures of ethanol with water. It was observed that the maximum experimental adsorption capacity, qm, exp, in water is up to 50 mg g-1 and decreases with increase in weight percentage of ethanol and NaCl and HCl concentration compared to that of water. Analysis of data using ARIAN model showed that MB adsorbs as monomer and dimer on the surface of agar. Binding constants of MB to agar were calculated using the Temkin isotherm. The process is exothermic in water and other solutions. The mean adsorption energy (E) value indicated binding of MB to agar is chemical adsorption. Kinetics of this interaction obeys from the pseudo-second-order model and diffusion of the MB molecules into the agar is the main rate-controlling step. PMID:22339759

  4. Individual based simulations of bacterial growth on agar plates

    NASA Astrophysics Data System (ADS)

    Ginovart, M.; López, D.; Valls, J.; Silbert, M.

    2002-03-01

    The individual based simulator, INDividual DIScrete SIMulations (INDISIM) has been used to study the behaviour of the growth of bacterial colonies on a finite dish. The simulations reproduce the qualitative trends of pattern formation that appear during the growth of Bacillus subtilis on an agar plate under different initial conditions of nutrient peptone concentration, the amount of agar on the plate, and the temperature. The simulations are carried out by imposing closed boundary conditions on a square lattice divided into square spatial cells. The simulator studies the temporal evolution of the bacterial population possible by setting rules of behaviour for each bacterium, such as its uptake, metabolism and reproduction, as well as rules for the medium in which the bacterial cells grow, such as concentration of nutrient particles and their diffusion. The determining factors that characterize the structure of the bacterial colony patterns in the presents simulations, are the initial concentrations of nutrient particles, that mimic the amount of peptone in the experiments, and the set of values for the microscopic diffusion parameter related, in the experiments, to the amount of the agar medium.

  5. 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. PMID:11472802

  6. 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. PMID:26355542

  7. 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

  8. A Rapid and Simple Bioassay Method for Herbicide Detection

    PubMed Central

    Li, Xiu-Qing; Ng, Alan; King, Russell; Durnford, Dion G.

    2008-01-01

    Chlamydomonas reinhardtii, a unicellular green alga, has been used in bioassay detection of a variety of toxic compounds such as pesticides and toxic metals, but mainly using liquid culture systems. In this study, an algal lawn—agar system for semi-quantitative bioassay of herbicidal activities has been developed. Sixteen different herbicides belonging to 11 different categories were applied to paper disks and placed on green alga lawns in Petri dishes. Presence of herbicide activities was indicated by clearing zones around the paper disks on the lawn 2–3 days after application. The different groups of herbicides induced clearing zones of variable size that depended on the amount, mode of action, and chemical properties of the herbicides applied to the paper disks. This simple, paper-disk-algal system may be used to detect the presence of herbicides in water samples and act as a quick and inexpensive semi-quantitative screening for assessing herbicide contamination. PMID:19578512

  9. 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)

  10. Bioassay for assessing marine contamination

    SciTech Connect

    Lapota, D.; Copeland, H.; Mastny, G.; Rosenberger, D.; Duckworth, D.

    1996-03-01

    The Qwiklite bioassay, developed by the laboratory at NCCOSC, is used as a biological tool to gauge the extent of environmental contamination. Some species of marine phytoplankton produce bioluminescence. The Qwiklite bioassay determines acute response and chronic effects of a wide variety of toxicants upon bioluminescent dinotlagellates by measuring their light output after exposure.

  11. BIOASSAY VESSEL FAILURE ANALYSIS

    SciTech Connect

    Vormelker, P

    2008-09-22

    Two high-pressure bioassay vessels failed at the Savannah River Site during a microwave heating process for biosample testing. Improper installation of the thermal shield in the first failure caused the vessel to burst during microwave heating. The second vessel failure is attributed to overpressurization during a test run. Vessel failure appeared to initiate in the mold parting line, the thinnest cross-section of the octagonal vessel. No material flaws were found in the vessel that would impair its structural performance. Content weight should be minimized to reduce operating temperature and pressure. Outer vessel life is dependent on actual temperature exposure. Since thermal aging of the vessels can be detrimental to their performance, it was recommended that the vessels be used for a limited number of cycles to be determined by additional testing.

  12. Evaluation and simplification of the assimilable organic carbon nutrient bioassay for bacterial growth in drinking water.

    PubMed

    Kaplan, L A; Bott, T L; Reasoner, D J

    1993-05-01

    A modified assimilable organic carbon (AOC) bioassay is proposed. We evaluated all aspects of the AOC bioassay technique, including inoculum, incubation water, bioassay vessel, and enumeration technique. Other concerns included eliminating the need to prepare organic carbon-free glassware and minimizing the risks of bacterial and organic carbon contamination. Borosilicate vials (40 ml) with Teflon-lined silicone septa are acceptable incubation vessels. Precleaned vials are commercially available, and the inoculum can be injected directly through the septa. Both bioassay organisms, Pseudomonas fluorescens P-17 and Spirillum sp. strain NOX, are available from the American Type Culture Collection and grow well on R2A agar, making this a convenient plating medium. Turbid raw waters need to be filtered prior to an AOC analysis. Glass fiber filters used with either a peristaltic pump or a syringe-type filter holder are recommended for this purpose. A sampling design that emphasizes replication of the highest experimental level, individual batch cultures, is the most efficacious way to reduce the total variance associated with the AOC bioassay. Quality control for the AOC bioassay includes an AOC blank and checks for organic carbon limitation and inhibition of the bioassay organisms. PMID:8517748

  13. Demonstrating Diffusion

    ERIC Educational Resources Information Center

    Foy, Barry G.

    1977-01-01

    Two demonstrations are described. Materials and instructions for demonstrating movement of molecules into cytoplasm using agar blocks, phenolphthalein, and sodium hydroxide are given. A simple method for demonstrating that the rate of diffusion of a gas is inversely proportional to its molecular weight is also presented. (AJ)

  14. 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. PMID:26596889

  15. Pigments of fly agaric (Amanita muscaria).

    PubMed

    Stintzing, Florian; Schliemann, Willibald

    2007-01-01

    The complex pigment pattern of fly agaric (Amanita muscaria) cap skins has been studied by LC-DAD and mass spectrometry. Among the betaxanthins the corresponding derivatives of serine, threonine, ethanolamine, alanine, Dopa, phenylalanine and tryptophan are reported for the first time to contribute to the pigment pattern of fly agarics. Betalamic acid, the chromophoric precursor of betaxanthins and betacyanins, muscaflavin and seco-dopas were also detected. Furthermore, the red-purple muscapurpurin and the red muscarubrin were tentatively assigned while further six betacyanin-like components could not be structurally allocated. Stability studies indicated a high susceptibility of pigment extracts to degradation which led to rapid colour loss thus rendering a complete characterization of betacyanin-like compounds impossible at present. Taking into account these difficulties the presented results may be a starting point for a comprehensive characterization of the pigment composition of fly agarics. PMID:18274277

  16. 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.

  17. Prostaglandins, bioassay and inflammation

    PubMed Central

    Flower, R J

    2006-01-01

    The formation of the British Pharmacological Society coincided almost exactly with a series of ground-breaking studies that ushered in an entirely new field of research – that of lipid mediator pharmacology. For many years following their chemical characterisation, lipids were considered only to be of dietary or structural importance. From the 1930s, all this changed – slowly at first and then more dramatically in the 1970s and 1980s with the emergence of the prostaglandins (PGs), the first intercellular mediators to be clearly derived from lipids, in a dynamic on-demand system. The PGs exhibit a wide range of biological activities that are still being evaluated and their properties underlie the action of one of the world's all-time favourite medicines, aspirin, as well as its more modern congeners. This paper traces the development of the PG field, with particular emphasis on the skilful utilisation of the twin techniques of bioassay and analytical chemistry by U.K. and Swedish scientists, and the intellectual interplay between them that led to the award of a joint Nobel Prize to the principal researchers in the PG field, half a century after the first discovery of these astonishingly versatile mediators. PMID:16402103

  18. Trace Amounts of Furan-2-Carboxylic Acids Determine the Quality of Solid Agar Plates for Bacterial Culture

    PubMed Central

    Hara, Shintaro; Isoda, Reika; Tahvanainen, Teemu; Hashidoko, Yasuyuki

    2012-01-01

    Background Many investigators have recognised that a significant proportion of environmental bacteria exist in a viable but non-culturable state on agar plates, and some researchers have also noticed that some of such bacteria clearly recover their growth on matrices other than agar. However, the reason why agar is unsuitable for the growth of some bacteria has not been addressed. Methodology/Principal Findings According to the guide of a bioassay for swarming inhibition, we identified 5-hydroxymethylfuran-2-carboxylic acid (5-HMFA) and furan-2-carboxylic acid (FA) as factors that inhibit bacterial swarming and likely inhibit extracellular polysaccharide production on agar. The furan-2-carboxylic acids 5-HMFA and FA effectively inhibited the swarming and swimming of several environmental bacteria at concentrations of 1.8 and 2.3 µg L−1 (13 and 21 nmol L−1), respectively, which are equivalent to the concentrations of these compounds in 0.3% agar. On Luria-Bertani (LB) plates containing 1.0% agar that had been previously washed with MeOH, a mixture of 5-HMFA and FA in amounts equivalent to their original concentrations in the unwashed agar repressed the swarming of Escherichia coli K12 strain W3110, a representative swarming bacterium. Conclusions/Significance Agar that contains trace amounts of 5-HMFA and FA inhibits the proliferation of some slow-growing or difficult-to-culture bacteria on the plates, but it is useful for single colony isolation due to the ease of identification of swarmable bacteria as the non-swarmed colonies. PMID:22848437

  19. 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

  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. Lecithin-agar assay for lecithinase antibodies in serum.

    PubMed

    Sibinovic, K H; Brown, F A; Pettigrew, K D; Vought, R L

    1971-01-01

    A technique for assay of lecithinase antibodies in serum was developed in this laboratory by using a lecithin-agar plate diffusion procedure based on a combination of described plate assays. Egg yolk lipoprotein composed primarily of lecithin was used as a substrate for reaction with free or non-neutralized lecithinase C after incubation of known amounts of lecithinase C with various dilutions of control and test sera. It was found that the size of the reaction zone was a function of enzyme concentration and inversely proportional to the antibody concentration. Accuracy and precision of the assay were determined. In addition, lecithinase antibody levels in sera from experimentally inoculated rats and rabbits and sera from randomly selected human patients were studied. PMID:4322282

  2. Lecithin-Agar Assay for Lecithinase Antibodies in Serum

    PubMed Central

    Sibinovic, Kyle H.; Brown, Freddie A.; Pettigrew, Karen D.; Vought, Robert L.

    1971-01-01

    A technique for assay of lecithinase antibodies in serum was developed in this laboratory by using a lecithin-agar plate diffusion procedure based on a combination of described plate assays. Egg yolk lipoprotein composed primarily of lecithin was used as a substrate for reaction with free or non-neutralized lecithinase C after incubation of known amounts of lecithinase C with various dilutions of control and test sera. It was found that the size of the reaction zone was a function of enzyme concentration and inversely proportional to the antibody concentration. Accuracy and precision of the assay were determined. In addition, lecithinase antibody levels in sera from experimentally inoculated rats and rabbits and sera from randomly selected human patients were studied. Images PMID:4322282

  3. Modelling larval movement data from individual bioassays.

    PubMed

    McLellan, Chris R; Worton, Bruce J; Deasy, William; Birch, A Nicholas E

    2015-05-01

    We consider modelling the movements of larvae using individual bioassays in which data are collected at a high-frequency rate of five observations per second. The aim is to characterize the behaviour of the larvae when exposed to attractant and repellent compounds. Mixtures of diffusion processes, as well as Hidden Markov models, are proposed as models of larval movement. These models account for directed and localized movements, and successfully distinguish between the behaviour of larvae exposed to attractant and repellent compounds. A simulation study illustrates the advantage of using a Hidden Markov model rather than a simpler mixture model. Practical aspects of model estimation and inference are considered on extensive data collected in a study of novel approaches for the management of cabbage root fly. PMID:25764283

  4. Primary Bioassay of Human Myeloma Stem Cells

    PubMed Central

    Hamburger, Anne; Salmon, Sydney E.

    1977-01-01

    The ability to clone primary tumors in soft agar has proven useful in the study of the kinetics and biological properties of tumor stem cells. We report the development of an in vitro assay which permits formation of colonies of human monoclonal plasma cells in soft agar. Colony growth has been observed from bone marrow aspirates from 75% of the 70 patients with multiple myeloma or related monoclonal disorders studied. Growth was induced with either 0.02 ml of human type O erythrocytes or 0.25 ml of medium conditioned by the adherent spleen cells of mineral oil-primed BALB/c mice. 5-500 colonies appeared after 2-3 wk in culture yielding a plating efficiency of 0.001-0.1%. The number of myeloma colonies was proportional to the number of cells plated between concentrations of 105-106 and back-extrapolated through zero, suggesting that colonies were clones derived from single myeloma stem cells. Morphological, histochemical, and functional criteria showed the colonies to consist of immature plasmablasts and mature plasma cells. 60-80% of cells picked from colonies contained intracytoplasmic monoclonal immunoglobulin. Colony growth was most easily achieved from the bone marrow cells of untreated patients or those in relapse. Only 50% of bone marrow samples from patients in remission were successfully cultured. Tritiated thymidine suicide studies provided evidence that for most myeloma patients, a very high proportion of myeloma colony-forming cells was actively in transit through the cell cycle. Velocity sedimentation at 1 g showed myeloma stem cells sedimented in a broad band with a peak at 13 mm/h. Antibody to granulocyte colony-stimulating factor did not reduce the number or size of the colonies. Increased numbers of myeloma colonies were seen when the marrow was depleted of colony-stimulating factor elaborating adherent cells before plating. This bioassay should prove useful in studying the in vitro biological behavior of certain bone marrow-derived (B

  5. Sediment bioassays with oyster larvae

    SciTech Connect

    Chapman, P.M.; Morgan, J.D.

    1983-10-01

    Tests with naturally-occurring sediments are rare and sediment testing methodology is not standardized. The authors present a simple methodology for undertaking sediment bioassays with oyster larvae, and present data from a recent study to prove the utility of this method.

  6. UNIFYING SCALER FOR BIOASSAY TESTS

    EPA Science Inventory

    An extensive set of interlaboratory root bioassay data was unified using centroids of individual tests as scalers. It is shown that the dose response obeys a first order differential equation with the constant of the equation related to the sensitivity of the dose response relati...

  7. 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

  8. Multiple screening of medicinal plants from Oaxaca, Mexico: ethnobotany and bioassays as a basis for phytochemical investigation.

    PubMed

    Frei, B; Heinrich, M; Bork, P M; Herrmann, D; Jaki, B; Kato, T; Kuhnt, M; Schmitt, J; Schühly, W; Volken, C; Sticher, O

    1998-05-01

    Based on ethnobotanical data collected among Zapotec Indians in Mexico, nine species traditionally applied to treat skin diseases and two species used to treat gastrointestinal disorders were subjected to several bioassays as further selection criteria for phytochemical investigation. Ten were active against at least one of the pathogenic and/or non-pathogenic bacteria and one against a non-pathogenic fungus in bioautographic TLC and agar diffusion tests. Cytotoxic/antitumor potential was found for one plant species with cell lines (KB, Caco-2) and for six with the brine shrimp assay. In the NF-κB- and the HET-CAM-test used to test for anti-inflammatory potential, two respectively one plant extract showed noteworthy activity. Furthermore, a potentially immunomodulating activity was investigated by evaluating the influence of extracts in various in vitro assays using murine and human lymphoid cells. In addition to the reported biological activities of the eleven plant species, comparisons of the ethnobotanical data and strategies for the selection for further phytochemical investigations are discussed. PMID:23195838

  9. 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

  10. 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. PMID:25842535

  11. Environmental monitoring using genetic bioassays

    SciTech Connect

    Lewtas, J.

    1989-01-01

    Environmental monitoring has evolved over the last ten years toward providing data more useful for exposure and risk assessment. The objective of many monitoring studies in the 1960s and 1970s was to monitor concentrations of pollutants including environmental mutagens at ambient locations, such as roof tops and in large bodies of water, where the pollutants would be well mixed and represent a homogeneous sample. In the 1980s, a number of studies focused on assessing the emission of mutagens from various sources. Now the emphasis has shifted to monitoring human exposure to environmental mutagens and to understanding which sources and factors lead to increased exposure and potential cancer risk. The chapter briefly reviews advances in genetic bioassay methods for environmental monitoring and focuses on approaches to integrating genetic bioassay methods with environmental-monitoring studies.

  12. 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.

  13. 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

  14. 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

  15. Automatic Surface Inoculation of Agar Trays1

    PubMed Central

    Wilkins, Judd R.; Mills, Stacey M.; Boykin, Elizabeth H.

    1972-01-01

    A machine is described which automatically inoculates a plastic tray containing agar media with a culture by use of either a conventional inoculating loop or a cotton swab. Isolated colonies were obtained with an inoculating loop when a heavy inoculum (109 cells/ml) was used or with a cotton swab when a light inoculum (ca. 104 cells/ml) was used. Trays containing combinations of differential or selective media were used to (i) separate mixtures of gram-positive and gram-negative bacteria, (ii) facilitate isolation of organisms from clinical specimens, and (iii) compare colony growth characteristics of pure cultures. The design of the machine is simple, it is easy to use, and it relieves the operator from the manual task of streaking cultures. Images PMID:16349943

  16. Poisoning with brown fly agaric, Amanita regalis.

    PubMed

    Elonen, E; Tarssanen, L; Härkönen, M

    1979-01-01

    Three patients ate different amounts of a common northern mushroom, brown fly agaric, Amanita regalis. All of them believed they had eaten delicious parasol mushrooms, Macrolepiota procera. The symptoms of poisoning began 1--2 hours after ingestion of the mushrooms. All the patients had marked gastrointestinal symptoms: nausea and heavy vomiting. Two had central nervous system manifestations and cholinergic symptoms: hallucinations, confusion, or loss of consciousness as well as copious salivation, or sweating. All patients recovered within 4--24 hours without any damage to liver, kidneys or central nervous system. It seems that cooking the mushrooms does not completely neutralize the toxic agents of Amanita regalis. The analysis of fried mushrooms shows that it may be possible to identify mushrooms reliably from the remains of a meal. PMID:760400

  17. Nanomaterial-Based Electrochemical Biosensors and Bioassays

    SciTech Connect

    Liu, Guodong; Mao, Xun; Gurung, Anant; Baloda, Meenu; Lin, Yuehe; He, Yuqing

    2010-08-31

    This book chapter summarizes the recent advance in nanomaterials for electrochemical biosensors and bioassays. Biofunctionalization of nanomaterials for biosensors fabrication and their biomedical applications are discussed.

  18. 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.

  19. Selected elements in fly agaric Amanita muscaria.

    PubMed

    Falandysz, J; Kunito, T; Kubota, R; Lipka, K; Mazur, A; Falandysz, Justyna J; Tanabe, S

    2007-09-01

    Concentrations of Ag, Al, Ba, Ca, Cd, Co, Cu, Cr, Cs, Fe, Ga, Hg, K, Mg, Mn, Mo, Na, Pb, Rb, Se, Sb, Sr, V, Tl and Zn have been determined in the whole fruiting bodies, as well as separately in caps and stalks, of fly agaric collected from three geographically distant sites in northern part of Poland. The elements were determined using ICP-MS, ICP-OES, HG-AAS and CV-AAS, respectively. For elements such as Al, Ba, Cr, Fe, Ga, Mo, Mn, Pb, Sb, Sr, Tl, and V concentrations were similar in the caps and stalks, respectively, and for K, Zn, Ag, Ca, Cd, Cu, Hg, Mg, Rb and Se were greater in the caps, while for Co, Cs and Na in the stalks. For Ag, Al, Ba, Ca, Cd, Co, Cr, Cs, Fe, Ga, Hg, Mn, Mo, Pb, Rb, Sb, Sr, Tl and V concentration in the caps showed spatial variations (P<0.05), while for Cu, K, Mg, Na, Se and Zn was independent of the site. The elements such as K with median or mean in the caps between 37,000 and 43,000 microg/g.dm and Mg with 920 and 1,100 microg/g dm were most abundant. Next, within median values range from approximately 100 to 500 microg/g dm were such as Ca, Fe and Al, and in descending order they followed by Rb (100-400 microg/g dm); V, Na, Zn (50-200 microg/g dm); Cu, Mn (10-50 microg/g dm); Cd (10-20 microg/g dm); Se (5 microg/g dm); Ba (<1-3); Cr, Ag, Pb, Sr (<1-2 microg/g dm); Cs, Co, Hg (<1-1 microg/g dm); Ga (<0.5), Sb, Mo and Tl (<0.1 microg/g dm). PMID:17849303

  20. Nanoparticle-Based Biosensors and Bioassays

    SciTech Connect

    Liu, Guodong; Wang, Jun; Lin, Yuehe; Wang, Joseph

    2007-10-11

    In this book chapter, we review the recent advances in nanoparticles based bioassay. The nanoparticles include quantum dots, silica nanoparticles and apoferritin nanoparticles. The new nanoparticles-based labels hold great promise for multiplex protein and DNA detection and for enhancing the sensitivity of other bioassays.

  1. Bioassays Based on Molecular Nanomechanics

    DOE PAGESBeta

    Majumdar, Arun

    2002-01-01

    Recent experiments have shown that when specific biomolecular interactions are confined to one surface of a microcantilever beam, changes in intermolecular nanomechanical forces provide sufficient differential torque to bend the cantilever beam. This has been used to detect single base pair mismatches during DNA hybridization, as well as prostate specific antigen (PSA) at concentrations and conditions that are clinically relevant for prostate cancer diagnosis. Since cantilever motion originates from free energy change induced by specific biomolecular binding, this technique is now offering a common platform for label-free quantitative analysis of protein-protein binding, DNA hybridization DNA-protein interactions, and in general receptor-ligandmore » interactions. Current work is focused on developing “universal microarrays” of microcantilever beams for high-throughput multiplexed bioassays.« less

  2. A bioaccumulation bioassay for freshwater sediments

    USGS Publications Warehouse

    Mac, Michael J.; Noguchi, George E.; Hesselberg, Robert J.; Edsall, Carol C.; Shoesmith, John A.; Bowker, James D.

    1990-01-01

    A laboratory bioassay is described for determining the bioavailability of contaminants from freshwater sediments. The bioassay consists of 10-d exposures to whole sediments under flow-through conditions. After testing five species, the fathead minnow (Pimephales promelas) and the earthworm (Lubricus terrestris) were recommended for use in the test. When the availability of polychlorinated biphenyls (PCBs), Hg and Zn from Great Lakes sediments was examined in laboratory exposures, only the PCBs were accumulated. A field validation study demonstrated that the magnitude of accumulation in laboratory exposures was similar to that in organisms caged in the field. A protocol is recommended for using the test as a standardized bioaccumulation bioassay.

  3. Simplified 48-hour IMVic test: an agar plate method.

    PubMed

    Powers, E M; Latt, T G

    1977-09-01

    An agar plate method was developed for the performance of the IMVic (indole, methyl red, Voges-Proskauer, and citrate) tests in lieu of the conventional tubed liquid media. By modifying the composition of the media and adding agar, a single "X"-compartmented petri dish was prepared containing all four IMVic test media. Ease of performance and simplification of the test were achieved by inoculating all four media simultaneously from a single colony (single inoculum) on eosin-methylene blue agar. Tests with 87 cultures, representing 7 genera in the family Enterobacteriaceae, were completed with typical (correct) IMVic patterns for all cultures within 48 h. Parallel tests with conventional media showed that the agar plate method was superior, more sensitive, faster, and simpler to perform, and less time was required to identify Escherichia coli by 72 h. PMID:334074

  4. Bioassay and characterization of soil microorganisms involved in the biodegradation of the fungicide, metalaxyl

    SciTech Connect

    Bailey, A.M.

    1985-01-01

    A sensitive bioassay was developed to detect low concentrations of metalaxyl in soils. The quantitative estimation of metalaxyl in soils was based on a significant positive relationship between the radial growth of Phytophthora boehmeriae and the log concentration of the fungicide in the agar. The isolate of P. boehmeriae was chosen for its sensitivity to metalaxyl as manifested in a linear growth response on cornmeal agar over a range of 2 to 30 ng/ml. The sensitivity and quantitative nature of the bioassay was confirmed by comparison with data obtained by using /sup 14/C-metalaxyl. Metabolism of metalaxyl was detected in three of five avocado soils that had repeated applications of the fungicide over 2-5 yr. The average disappearance of metalaxyl was 28 days, and in the most active soils was 14 days. The composition and level of the microbial populations of soils, either active or inactive in the breakdown of metalaxyl, did not differ. Fungal and bacterial microflora recovered from these two soils by use of either selective media or filtration techniques were capable of metabolizing metalaxyl over a 45-day period.

  5. Two-generation saccharin bioassays.

    PubMed

    Arnold, D L

    1983-04-01

    The controversy regarding the safety of saccharin for human consumption started shortly after its discovery over 100 years ago and has yet to subside appreciably. The consumption of saccharin, particularly in North America, began to escalate when the U.S. Food and Drug Administration set new standards of identity which allowed foods containing artificial sweeteners to be promoted as "nonnutritive" or "noncaloric" sweeteners for use by the general public. In 1969, when cyclamates were banned, at least 10 single-generation feeding studies were undertaken with saccharin to more accurately assess the potential toxicological consequences resulting from the anticipated increase in its consumption. None of these studies resulted in any overt regulatory action. Subsequently, the introduction of the two-generation chronic toxicity/carcinogenicity bioassay added a new tool to the toxicologist's arsenal. Three two-generation studies using saccharin have since been conducted. The results from these studies clearly show that when rats were exposed to diets containing 5 or 7.5% sodium saccharin from the time of conception to death, an increased frequency of urinary bladder cancers was found, predominantly in the males. While some study results suggested that impurities in commercial saccharin or the presence of urinary tract calculi may have been responsible for the observed bladder tumors, it now appears that these possibilities are highly unlikely. The mechanism by which saccharin elicited the bladder tumors using the two-generation experiment has not been ascertained. PMID:6347682

  6. Bioassays for Monitoring Insecticide Resistance

    PubMed Central

    Miller, Audra L.E.; Tindall, Kelly; Leonard, B. Rogers

    2010-01-01

    Pest resistance to pesticides is an increasing problem because pesticides are an integral part of high-yielding production agriculture. When few products are labeled for an individual pest within a particular crop system, chemical control options are limited. Therefore, the same product(s) are used repeatedly and continual selection pressure is placed on the target pest. There are both financial and environmental costs associated with the development of resistant populations. The cost of pesticide resistance has been estimated at approximately $ 1.5 billion annually in the United States. This paper will describe protocols, currently used to monitor arthropod (specifically insects) populations for the development of resistance. The adult vial test is used to measure the toxicity to contact insecticides and a modification of this test is used for plant-systemic insecticides. In these bioassays, insects are exposed to technical grade insecticide and responses (mortality) recorded at a specific post-exposure interval. The mortality data are subjected to Log Dose probit analysis to generate estimates of a lethal concentration that provides mortality to 50% (LC50) of the target populations and a series of confidence limits (CL's) as estimates of data variability. When these data are collected for a range of insecticide-susceptible populations, the LC50 can be used as baseline data for future monitoring purposes. After populations have been exposed to products, the results can be compared to a previously determined LC50 using the same methodology. PMID:21248689

  7. PHOXOCEPHALID AMPHIPOD BIOASSAY FOR MARINE SEDIMENT TOXICITY

    EPA Science Inventory

    The relative toxicity of marine sediment can be accurately determined through acute, static bioassays with the phoxocepalid amphipod Repoxynius abronius. Mortality and sublethal effects on emergence from sediment and reburial behavior are determined after ten day exposure in 1-L ...

  8. Characterization of agar/soy protein biocomposite films: Effect of agar on the extruded pellets and compression moulded films.

    PubMed

    Garrido, T; Etxabide, A; Guerrero, P; de la Caba, K

    2016-10-20

    Agar/soy protein biocomposite films were successfully processed by extrusion and compression moulding, obtaining transparent and homogeneous films. The conformational changes occurred during the extrusion process and the effect of agar on the final properties were analyzed. As shown by differential scanning calorimetry (DSC) and specific mechanical energy (SME) values, during the extrusion process protein denatured and unfolded protein chains could interact with agar. These interactions were analyzed by Fourier transform infrared spectroscopy (FTIR) and the secondary structure was determined from the amide I band. Those interactions were supported by the decrease of film solubility. Furthermore, the good compatibility between agar and soy protein was confirmed by the images from scanning electron microscopy (SEM). PMID:27474583

  9. Bioassay criteria for environmental restoration workers

    SciTech Connect

    Carbaugh, E.H.; Bihl, D.E.

    1993-01-01

    Environmental restoration (ER) work at the U. S. Department of Energy Hanford Site posed questions concerning when to perform bioassay monitoring of workers for potential intakes of radioactivity. Application of criteria originally developed for use inside radionuclide processing facilities to ER work resulted in overly restrictive bioassay requirements. ER work typically involves site characterization or, excavating large quantities of potentially contaminated soil, rather than working with concentrated quantities of radioactivity as in a processing facility. An improved approach, tailored to ER work, provided soil contamination concentrations above which worker bioassay would be required. Soil concentrations were derived assuming acute or chronic intakes of 2% of an Annual Limit on Intake (ALI), or a potential committed effective dose equivalent of 100 mrem, and conservative dust loading of air from the work. When planning ER work, the anticipated soil concentration and corresponding need for bioassay could be estimated from work-site historical records. Once site work commenced, soil sampling and work-place surveys could be used to determine bioassay needs. This approach substantially reduced the required number of bioassay samples with corresponding reductions in analytical costs, schedules, and more flexible work-force management. (Work supported by the US Department of Energy under contract DOE-AC06-76RLO 1830.)

  10. A Colorimetric Bioassay for Perchlorate

    NASA Astrophysics Data System (ADS)

    Heinnickel, M. L.; Smith, S.; Coates, J. D.

    2007-12-01

    Recognition of perchlorate (ClO4-) as a widespread contaminant across the United States and its potential adverse affects towards human health has motivated the EPA to place ClO4- on its contaminant candidate list for drinking water supplies. While a federal MCL has not yet been set, a recommended public health goal of 1 ppb (μg.L-1) was established by the US EPA in 2002. To date, methods of detection require use of sensitive ion chromatographic equipment that are expensive, time consuming, and require highly trained personnel for use. Our studies are focused on the development of a highly sensitive, simple, and robust colorimetric bioassay based on the primary enzyme involved in microbial ClO4- reduction, the perchlorate reductase (Pcr). A previously published assay used reduced methyl viologen (MV, the dye is reduced with sodium hydrosulfite) as an electron donor to demonstrate Pcr activity. The assay directly correlates the amount of MV oxidized with the amount of ClO4- reduced by assuming a transfer of four electrons. To test this assumption, we compared actual concentrations of MV oxidized to ClO4- reduced in this assay. ClO4- concentrations were determined using a Dionex ICS-500 ion chromatography system, while MV concentrations were determined using a standard curve generated at 578 nm. Comparisons between the two revealed that twelve molecules of MV were oxidized for each molecule of ClO4- reduced. The oxidation of these additional eight MV molecules is explained by the interaction of the dye with chlorite (the product of the Pcr reaction) and other contaminants that could be present in the enzyme prep. This unsettling result indicated this assay would be problematic for the detection of ClO4- in soil, which has many chemicals that could react with MV. To improve upon this assay, we have tried to reduce ClO4- using less reactive dyes and reductants. The reductants ascorbic acid, NADH, and dithiothreitol drive Pcr catalyzed ClO4- reduction, however, they

  11. Hyperspectral imaging for detecting pathogens grown on agar plates

    NASA Astrophysics Data System (ADS)

    Yoon, Seung Chul; Lawrence, Kurt C.; Siragusa, Gregory R.; Line, John E.; Park, Bosoon; Windham, William R.

    2007-09-01

    This paper is concerned with the development of a hyperspectral imaging technique for detecting and identifying one of the most common foodborne pathogens, Campylobacter. Direct plating using agars is an effective tool for laboratory tests and analyses of microorganisms. The morphology (size, growth pattern, color, etc.) of colonies grown on agar plates has been widely used to tentatively differentiate organisms. However, it is sometimes difficult to differentiate target organisms like Campylobacters from other contaminants grown together on the same agar plates. A hyperspectral imaging system operating at the visible and near infrared (VNIR) spectral region from 400 nm to 900 nm was set up to measure spectral signatures of 17 different Campylobacter and non-Campylobacter subspecies. Protocols for culturing, imaging samples and for calibrating measured data were developed. The VNIR spectral library of all 17 organisms commonly encountered in poultry was established from calibrated hyperspectral images. A classification algorithm was developed to locate and identify Campylobacters, non-Campylobacter contaminants, and background agars with 99.29% accuracy. This research has a potential to be expanded to detect other pathogens grown on agar media.

  12. 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

  13. Effect of BiTek agar on lysostaphin susceptibility of staphylococci.

    PubMed Central

    Langlois, B E; Dawson, K; Akers, K

    1990-01-01

    Staphylococci which were considered to be lysostaphin susceptible on P agar containing Bacto-Agar showed different degrees of resistance to lysostaphin when tested on P agar made with BiTek agar. As a result, lysostaphin-susceptible strains were misidentified as lysostaphin-resistant strains. Images PMID:2254432

  14. Effects of metals in in vitro bioassays.

    PubMed Central

    Sirover, M A

    1981-01-01

    The capacity of in vitro bioassays to detect the potential carcinogenicity of metal compounds is reviewed. The in vitro bioassays discussed include: bacterial reversion analysis to determine the capacity of metal salts to revert Salmonella typhimurium histidine auxotrophs or to revert Escherichia coli WP 2 tryp- to tryptophan prototrophy; examination of the ability of metal salts to preferentially inhibit cell growth in Bacillus subtilis cells deficient in DNA repair pathways; determination of the ability of metal salts to induce resistance to base analogs in mammalian cells; the capacity of metal salts to enhance viral transformation of mammalian cells or to transform cells in the absence of virus; and the ability of metal salts to induce chromosomal aberrations in mammalian cells. Using each of these in vitro bioassays, diverse metal compounds have been identified as potential carcinogens. Furthermore, the use of different compounds of a specific metal may allow a determination of the valence which may be required for carcinogenesis. PMID:7023930

  15. Poultry litter toxicity comparison from various bioassays

    SciTech Connect

    Gupta, G.; Kelly, P. )

    1992-01-01

    Poultry litter contains many toxic chemicals including Cu, As, Pb, Cd, Hg, Se and PCBs. Poultry litter leachate has been shown to be more toxic to marine luminescent organisms (Photobacterium phosphoreum) than other farm animal manures. A comparison of toxicity of the poultry litter leachate was undertaken using various bioassays. The EC{sub 50} (or LC{sub 50}) value for the leachate with the Microtox and Daphnia bioassays was 2.9 g/L/ Nitrobacter and Pseudomonas bioassays were not useful in determining the leachate toxicity because of the nutritional properties of the litter. Poultry litter leachate was found to be mutagenic to strains TA 97, TA 98, TA 100 and TA 102 using the Ames Test.

  16. RECOMMENDATIONS FOR UO3 PLANT BIOASSAY

    SciTech Connect

    Carbaugh, Eugene H.

    2010-07-12

    Alternative urine bioassay programs are described for application with decontamination and decommissioning activities at the Hanford UO3 Plant. The alternatives are based on quarterly or monthly urine bioassay for recycled uranium, assuming multiple acute inhalation intakes of recycled uranium occurring over a year. The inhalations are assumed to be 5µm AMAD particles of 80% absorption type F and 20% absorption type M. Screening levels, expressed as daily uranium mass excretion rates in urine, and the actions associated with these levels are provided for both quarterly and monthly sampling frequencies.

  17. 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

  18. 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

  19. 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.

  20. Hyperspectral Imaging for Detecting Pathogens Grown on Agar Plates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper is concerned with the development of a hyperspectral imaging technique for detecting and identifying one of the most common foodborne pathogens, Campylobacter. Direct plating using agars is an effective tool for laboratory tests and analyses of microorganisms. The morphology (size, growth...

  1. 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)

  2. 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 §...

  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. 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 §...

  5. 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 §...

  6. 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 §...

  7. 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...

  8. BIOASSAY-DIRECTED CHEMICAL ANALYSIS IN ENVIRONMENTAL RESEARCH

    EPA Science Inventory

    The use of short-term bioassay tests in conjunction with analytical measurements, constitute a powerful tool for identifying important environmental contaminants. The authors have coined the terminology 'bioassay directed chemical analysis' to best describe this marriage of analy...

  9. Micro-organism distribution sampling for bioassays

    NASA Technical Reports Server (NTRS)

    Nelson, B. A.

    1975-01-01

    Purpose of sampling distribution is to characterize sample-to-sample variation so statistical tests may be applied, to estimate error due to sampling (confidence limits) and to evaluate observed differences between samples. Distribution could be used for bioassays taken in hospitals, breweries, food-processing plants, and pharmaceutical plants.

  10. EDC BIOASSAYS FOR RISK MANAGEMENT PROJECTS

    EPA Science Inventory

    Overall goal for this research is to develop 3 bioassays for use in EDC projects across NRMRL (estrogenic, androgenic and thyroid assays). Currently, research is focused on estrogenic assays. A literature search was conducted to identify potential assays. The Yeast Estrogen Sc...

  11. 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. PMID:25126968

  12. 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.

  13. 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

  14. Brine Shrimp Bioassays: A Useful Technique in Biological Investigations

    ERIC Educational Resources Information Center

    Rice, Stanley A.; Maness, Ian B.

    2004-01-01

    A technique to measure the potency of leaf compounds against herbivores with the use of a bioassay is described. Bioassays are useful in classes where students have career plans like medicine in which bioassays can be used as tools for screening plants for possible medicinal potency.

  15. BIOASSAY-DIRECTED FRACTIONATION OF ORGANIC CONTAMINANTS IN AN ESTUARINE SEDIMENT USING THE NEW MUTAGENIC BIOASSAY, MUTATOX

    EPA Science Inventory

    Bioassay-directed fractionation of organic compounds was performed on an organic solvent extract of a contaminated estuarine sediment from Black Rock Harbor, Connecticut, using the new mutagenic bioassay, Mutatox-. hemical fractionation methods of the sediment extract included si...

  16. Comparison of chromogenic Biolog Rainbow agar Shigella/Aeromonas with xylose lysine desoxycholate agar for isolation and detection of Shigella spp. from foods.

    PubMed

    Zhang, Guodong; Lampel, Keith A

    2010-08-01

    Shigella outbreaks are widely reported throughout the world. However, it remains a challenge to isolate Shigella spp. from foods by using conventional microbiological media. The main objective of this study was to determine the effectiveness of a novel chromogenic medium, Rainbow agar Shigella/Aeromonas (Rainbow agar), for the isolation and detection of Shigella spp. in foods. All four Shigella species, S. sonnei, S. flexneri, S. dysenteriae, and S. boydii, were studied. Rainbow agar was compared with tryptic soy agar, xylose lysine desoxycholate agar (XLD), and Salmonella Shigella agar (SSA) for enumeration of Shigella spp. in pure culture. This chromogenic agar and XLD were also used to isolate Shigella spp. in artificially contaminated foods (4.8 log CFU/g of food), including lettuce, parsley, cilantro, spinach, potato salad, and shrimp. The inhibitory effect on Shigella growth by Rainbow agar was between that of XLD and SSA. All vegetables studied showed a moderately high background microflora on XLD and Rainbow agar. With artificially inoculated produce, Rainbow agar recovered about 1 to 2 log CFU more S. sonnei, S. dysenteriae, and S. boydii per g of food than did XLD. For potato salad and shrimp, which had low background microflora on Rainbow agar, Rainbow agar was slightly better in recovering Shigella spp. than XLD was in most cases. However, we found that the addition of streptomycin (6.25 mg/liter) to Rainbow agar could facilitate the isolation of Shigella in vegetables tested. In conclusion, Rainbow agar was a much more effective medium than was XLD for the isolation of Shigella spp. from foods. PMID:20819355

  17. An agar gel enzyme assay (AGEA) for simple detection of Salmonella enteritidis antibodies in chicken sera.

    PubMed

    Kim, C J; Nagaraja, K V

    1991-01-01

    An agar gel enzyme assay (AGEA) was developed for the detection of antibodies to Salmonella enteritidis (SE). The assay was based on the ability of antibodies to diffuse through an agar gel and react with antigen coated on a polystyrene surface. The antigen-antibody reaction was then made visible by applying an enzyme-conjugated anti-immunoglobulin and the addition, subsequently, of a substrate-containing gel. The color change in circular zones was taken as the indication for the presence of antibodies. The present investigation reports identification of an antigen specific for SE and its use in the development of a relatively simple AGEA procedure. The results of AGEA were compared with those of conventional microagglutination (MA) test and serum plate (SP) test. The percentage agreement between MA and AGEA in positive serum sample was found to be 94.4%, and in negative serum samples it was found to be 88.8%. The present results suggest that the AGEA could be a very useful screening test for the detection of SE antibodies because the assay is inexpensive, specific and simple to perform without much equipment, and give results within a 3-hr period. PMID:1832368

  18. 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.

  19. 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

  20. Three-dimensional characterization of bacterial microcolonies on solid agar-based culture media.

    PubMed

    Drazek, Laurent; Tournoud, Maud; Derepas, Frédéric; Guicherd, Maryse; Mahé, Pierre; Pinston, Frédéric; Veyrieras, Jean-Baptiste; Chatellier, Sonia

    2015-02-01

    For the last century, in vitro diagnostic process in microbiology has mainly relied on the growth of bacteria on the surface of a solid agar medium. Nevertheless, few studies focused in the past on the dynamics of microcolonies growth on agar surface before 8 to 10h of incubation. In this article, chromatic confocal microscopy has been applied to characterize the early development of a bacterial colony. This technology relies on a differential focusing depth of the white light. It allows one to fully measure the tridimensional shape of microcolonies more quickly than classical confocal microscopy but with the same spatial resolution. Placing the device in an incubator, the method was able to individually track colonies growing on an agar plate, and to follow the evolution of their surface or volume. Using an appropriate statistical modeling framework, for a given microorganism, the doubling time has been estimated for each individual colony, as well as its variability between colonies, both within and between agar plates. A proof of concept led on four bacterial strains of four distinct species demonstrated the feasibility and the interest of the approach. It showed in particular that doubling times derived from early tri-dimensional measurements on microcolonies differed from classical measurements in micro-dilutions based on optical diffusion. Such a precise characterization of the tri-dimensional shape of microcolonies in their late-lag to early-exponential phase could be beneficial in terms of in vitro diagnostics. Indeed, real-time monitoring of the biomass available in a colony could allow to run well established microbial identification workflows like, for instance, MALDI-TOF mass-spectrometry, as soon as a sufficient quantity of material is available, thereby reducing the time needed to provide a diagnostic. Moreover, as done for pre-identification of macro-colonies, morphological indicators such as three-dimensional growth profiles derived from

  1. 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

  2. Antimicrobial Disk Susceptibility Testing of Leptospira spp. Using Leptospira Vanaporn Wuthiekanun (LVW) Agar.

    PubMed

    Wuthiekanun, Vanaporn; Amornchai, Premjit; Langla, Sayan; White, Nicholas J; Day, Nicholas P J; Limmathurotsakul, Direk; Peacock, Sharon J

    2015-08-01

    Leptospira Vanaporn Wuthiekanun (LVW) agar was used to develop a disk diffusion assay for Leptospira spp. Ten pathogenic Leptospira isolates were tested, all of which were susceptible to 17 antimicrobial agents (amoxicillin/clavulanic acid, amoxicillin, azithromycin, cefoxitin, ceftazidime, ceftriaxone, chloramphenicol, ciprofloxacin, clindamycin, doripenem, doxycycline, gentamicin, linezolid, nitrofurantoin, penicillin, piperacillin/tazobactam, and tetracycline). All 10 isolates had no zone of growth inhibition for four antimicrobials (fosfomycin, nalidixic acid, rifampicin, and trimethoprim/sulfamethoxazole). Of the ten Leptospira, seven had a growth inhibition zone of ≤ 21 mm for aztreonam, the zone diameter susceptibility break point for Enterobacteriaceae. This assay could find utility as a simple screening method during the epidemiological surveillance of antimicrobial resistance in Leptospira spp. PMID:26055750

  3. Molecular Diffusion Coefficients: Experimental Determination and Demonstration.

    ERIC Educational Resources Information Center

    Fate, Gwendolyn; Lynn, David G.

    1990-01-01

    Presented are laboratory methods which allow the demonstration and determination of the diffusion coefficients of compounds ranging in size from water to small proteins. Included are the procedures involving the use of a spectrometer, UV cell, triterated agar, and oxygen diffusion. Results including quantification are described. (CW)

  4. Bioassaying for ozone with pollen systems

    SciTech Connect

    Feder, W.A.

    1981-01-01

    Sensitivity to ozone of pollen germinating in vitro is closely correlated with ozone sensitivity of the pollen parent. Ozone-sensitive and tolerant pollen populations have been identified in tobacco, petunia, and tomato cultivars. The rate of tube elongation can be reversibly slowed or stopped by exposure to low concentrations of ozone. The performance of selected pollen populations can then be used to bioassay ozone in ambient air by introducing the air sample into a growth chamber where ozone-sensitive pollen in growing. Year-round pollen producion can be achieved in the greenhouse. Harvested pollen can be tested, packaged, and transported to user facilities without loss of vigor. Pollen populations are inexpensive to produce, respond reliably, and are simple to use as a bioassay for air quality.

  5. Bioassay Labels Based on Apoferritin Nanovehicles

    SciTech Connect

    Liu, Guodong; Wang, Jun; Lea, Alan S.; Lin, Yuehe

    2006-09-04

    Here we report a nanoparticle label based on apoferritin nanovehicle loaded internally with markers for sensitive electrochemical DNA detection. The central cavity structure, the dissociation and reconstitute properties at different pHs of apoferritin provide a facile method to load and release markers. Hexacynoferrate(III) was used as model marker to load into the cavity of apoferritin protein cage. The loaded nanoparticle surface was functionalized with amino-modified DNA probe. Electrochemical DNA hybridization assay based on the hexacynoferrate loaded apoferritin nanovehicle could detect 23 atmol DNA targets in 50 ul sample solution. The concept could be readily extended to load other redox and fluorescence markers for bioassay applications. The new nanoparticle labels hold great promise for multi-target detection (in connection to nanoparticles loaded with different markers) and for enhancing the sensitivity of other bioassays.

  6. Perspectives in avoidance-preference bioassays

    SciTech Connect

    Steele, C.W.; Taylor, D.H.; Strickler-Shaw, S.

    1996-12-31

    Although behavioral endpoints are used in hazard assessment, establishment of water quality criteria and assessment of a contaminant`s hazard to aquatic life rely primarily on standard acute and chronic toxicity tests. Sublethal effects of pollutants should, however, be of major concern because more organisms experience sublethal rather than acutely or chronically lethal exposures of contaminants. The avoidance-preference approach to behavioral bioassays is very useful in screening pollutants for which the mechanisms of perception or response are largely unknown. The underlying philosophy of these studies is that an animal which perceives a chemical can be attracted or repulsed by it. No response is frequently assumed to indicate lack of perception. All three responses have broad ecological implications. The authors discuss the conditions required for performing avoidance-preference bioassays, as well as their sensitivities, advantages, and limitations. In this regard, a comparative approach is used in examining the results of avoidance-preference bioassays with zebrafish in two different apparatuses. Finally, they compare the results of avoidance-preference studies with other measures of the behavioral toxicity of lead to tadpoles.

  7. 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

  8. 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...

  9. 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

  10. 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

  11. Choline chloride based ionic liquid analogues as tool for the fabrication of agar films with improved mechanical properties.

    PubMed

    Sousa, Ana M M; Souza, Hiléia K S; Latona, Nicholas; Liu, Cheng-Kung; Gonçalves, Maria P; Liu, LinShu

    2014-10-13

    In the present paper, we test the suitability of ChCl/urea (DES-U) and ChCl/glycerol (DES-G) eutectic mixtures, each one prepared at 1:2 molar ratio, for the production of agar films. A three-step process is proposed: pre-solubilization of polymer in DES followed by compression-molding and subsequent drying. The mechanical properties, water resistance and microstructure of the films were evaluated at different polymer concentrations (i.e. 2-6%, w/w). DES-U showed by far, the best film forming ability. Agreeing with the diffusion and SEM data, films with the best mechanical properties were found at the lowest and highest agar concentrations (tensile strengths of 24.2-42 MPa and elongations of 15.4-38.9%). The water sorption and contact angle studies suggested increased hydrophilicity for the film containing the lowest concentration of agar. The use of choline chloride based ionic liquid analogues as solvent and plasticizer might be a promising tool for the development of new non-aqueous materials based on seaweed polysaccharides. PMID:25037344

  12. In-situ bioassays using caged bivalves

    SciTech Connect

    Salazar, M.H.; Salazar, S.M.

    1995-12-31

    It is important to make the distinction between chemical measurements to assess bioaccumulation potential versus biological measurements to assess potential bioeffects because bioaccumulation is not a bioeffect. Caging provides a unique opportunity to make synoptic measurements of each and facilitates making these measurements over space and time. Measuring bioaccumulation in resident and transplanted bivalves has probably been the most frequently used form of an in-situ bioassay because bivalves concentrate chemicals in their tissues. They are also easy to collect, cage, and measure. The authors have refined bivalve bioassay methods by minimizing the size range of test animals, making repetitive measurements of the same individuals, and standardizing test protocols for a variety of applications. They are now attempting to standardize criteria for accepting and interpreting data in the same way that laboratory bioassays have been standardized. Growth measurements can serve two purposes in this assessment strategy: (1) An integrated biological response endpoint that is easily quantifiable and with significance to the population, and (2) A means of calibrating bioaccumulation by assessing the relative health and physiological state of tissues that have accumulated the chemicals. In general, the authors have found the highest bioconcentration factors associated with the highest growth rates, the highest concentrations ({micro}g/g) of chemicals in juvenile mussels, and the highest chemical content ({micro}g/animal) in adult mussels. Without accounting for possible dilution of chemical concentrations by tissue growth or magnification through degrowth, contaminant concentrations can be misleading. Examples are provided for the Sudbury River in Massachusetts (Elliptio complanata), San Diego Bay (Mytilus galloprovincialis), and the Harbor Island Superfund Site in Puget Sound (Mytilus trossulus).

  13. 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

  14. 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. PMID:21207092

  15. A Multichannel Bioluminescence Determination Platform for Bioassays.

    PubMed

    Kim, Sung-Bae; Naganawa, Ryuichi

    2016-01-01

    The present protocol introduces a multichannel bioluminescence determination platform allowing a high sample throughput determination of weak bioluminescence with reduced standard deviations. The platform is designed to carry a multichannel conveyer, an optical filter, and a mirror cap. The platform enables us to near-simultaneously determine ligands in multiple samples without the replacement of the sample tubes. Furthermore, the optical filters beneath the multichannel conveyer are designed to easily discriminate colors during assays. This optical system provides excellent time- and labor-efficiency to users during bioassays. PMID:27424912

  16. 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

  17. Urine sample collection protocols for bioassay samples

    SciTech Connect

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject`s body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  18. Urine sample collection protocols for bioassay samples

    SciTech Connect

    MacLellan, J.A.; McFadden, K.M.

    1992-11-01

    In vitro radiobioassay analyses are used to measure the amount of radioactive material excreted by personnel exposed to the potential intake of radioactive material. The analytical results are then used with various metabolic models to estimate the amount of radioactive material in the subject's body and the original intake of radioactive material. Proper application of these metabolic models requires knowledge of the excretion period. It is normal practice to design the bioassay program based on a 24-hour excretion sample. The Hanford bioassay program simulates a total 24-hour urine excretion sample with urine collection periods lasting from one-half hour before retiring to one-half hour after rising on two consecutive days. Urine passed during the specified periods is collected in three 1-L bottles. Because the daily excretion volume given in Publication 23 of the International Commission on Radiological Protection (ICRP 1975, p. 354) for Reference Man is 1.4 L, it was proposed to use only two 1-L bottles as a cost-saving measure. This raised the broader question of what should be the design capacity of a 24-hour urine sample kit.

  19. 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. PMID:24211606

  20. 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. PMID:27109059

  1. Aspirator Gun for High-Throughput Mosquito Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We describe an innovative aspirator gun designed to transfer anaesthetized mosquitoes directly into glass bioassay tubes. The gun has been used for thousands of transfers with extremely low associated mortality and is the central component of a high-throughput bioassay system. The gun is constructed...

  2. COLLECTION, CHEMICAL FRACTIONATION, AND MUTAGENICITY BIOASSAY OF AMBIENT AIR PARTICULATE

    EPA Science Inventory

    The influence of industrialization and consequent increased concentration of urban particulate matter on the incidence of cancer has long been a concern. The first bioassays used to evaluate complex ambient air samples were whole-animal carcinogenesis bioassays. In these studies,...

  3. Aspirator gun for high-throughput mosquito bioassays.

    PubMed

    Aldridge, Robert L; Wynn, W Wayne; Britch, Seth C; Linthicum, Kenneth J

    2012-03-01

    We describe an innovative aspirator gun designed to transfer individual anesthetized mosquitoes directly into glass bioassay tubes. The gun has been used for thousands of transfers with extremely low associated mortality and is the central component of a high-throughput bioassay system. The gun is constructed using readily obtainable materials and can be modified for a range of insects. PMID:22533090

  4. Bioassay vs. Air Sampling: Practical Guidance and Experience at Hanford

    SciTech Connect

    Carbaugh, Eugene H.; Carlson, Eric W.; Hill, Robin L.

    2004-02-08

    The Hanford Site has implemented a policy to guide in determining whether air sampling data or special fecal bioassay data are more appropriate for determining doses of record for low-level plutonium exposures. The basis for the policy and four years of experience in comparing DAC-hours exposure with bioassay-based dosimetry is discussed.

  5. Signal Amplification of Bioassay Using Zinc Nanomaterials

    NASA Astrophysics Data System (ADS)

    Cowles, Chad L.

    An emerging trend in the analytical detection sciences is the employment of nanomaterials for bioassay signal transduction to identify analytes critical to public health. These nanomaterials have been specifically investigated for applications which require identification of trace levels of cells, proteins, or other molecules that can have broad ranging impacts to human health in fields such as clinical diagnostics, environmental monitoring, food and drink control, and the prevention of bioterrorism. Oftentimes these nanoparticle-based signal transduction or amplification approaches offer distinct advantages over conventional methods such as increased sensitivity, rapidity, or stability. The biological application of nanoparticles however, does suffer from drawbacks that have limited more widespread adoption of these techniques. Some of these drawbacks are, high cost and toxicity, arduous synthesis methods, functionalization and bioconjugation challenges, and laboratory disposal and environmental hazard issues, all of which have impeded the progression of this technology in some way or another. This work aims at developing novel techniques that offer solutions to a number of these hurdles through the development of new nanoparticle-based signal transduction approaches and the description of a previously undescribed nanomaterial. Zinc-based nanomaterials offer the opportunity to overcome some of the limitations that are encountered when other nanomaterials are employed for bioassay signal transduction. On the other hand, the biological application of zinc nanomaterials has been difficult because in general their fluorescence is in the blue range and the reported quantum yields are usually too low for highly sensitive applications. The advantages of using zinc nanomaterials for biological applications, such as reduced toxicity, simple synthesis, low cost, and straightforward functionalization strategies contribute to the research interest in their application as

  6. Circular Bioassay Platforms for Applications in Microwave-Accelerated Techniques

    PubMed Central

    Mohammed, Muzaffer; Clement, Travis C.; Aslan, Kadir

    2014-01-01

    In this paper, we present the design of four different circular bioassay platforms, which are suitable for homogeneous microwave heating, using theoretical calculations (i.e., COMSOL™ multiphysics software). Circular bioassay platforms are constructed from poly(methyl methacrylate) (PMMA) for optical transparency between 400–800 nm, has multiple sample capacity (12, 16, 19 and 21 wells) and modified with silver nanoparticle films (SNFs) to be used in microwave-accelerated bioassays (MABs). In addition, a small monomode microwave cavity, which can be operated with an external microwave generator (100 W), for use with the bioassay platforms in MABs is also developed. Our design parameters for the circular bioassay platforms and monomode microwave cavity during microwave heating were: (i) temperature profiles, (ii) electric field distributions, (iii) location of the circular bioassay platforms inside the microwave cavity, and (iv) design and number of wells on the circular bioassay platforms. We have also carried out additional simulations to assess the use of circular bioassay platforms in a conventional kitchen microwave oven (e.g., 900 W). Our results show that the location of the circular bioassay platforms in the microwave cavity was predicted to have a significant effect on the homogeneous heating of these platforms. The 21-well circular bioassay platform design in our monomode microwave cavity was predicted to offer a homogeneous heating pattern, where inter-well temperature was observed to be in between 23.72–24.13°C and intra-well temperature difference was less than 0.21°C for 60 seconds of microwave heating, which was also verified experimentally. PMID:25568813

  7. Comparison of ChromID Agar and Clostridium difficile Selective Agar for Effective Isolation of C. difficile from Stool Specimens

    PubMed Central

    Lee, Eun Joo

    2014-01-01

    Background ChromID Clostridium difficile agar (IDCd; bioMérieux SA, France) is a recently developed chromogenic medium for rapid and specific isolation of C. difficile. We compared the performance of IDCd with that of Clostridium difficile Selective Agar (CDSA). Methods A total of 530 fresh stool specimens were collected from patients with clinical signs compatible with C. difficile infection, and cultures for C. difficile were performed on IDCd and CDSA. C. difficile colonies were identified by spore staining, odor, use of an ANI identification test kit (bioMérieux SA), and multiplex PCR for tcdA, tcdB, and tpi. Results The concordance rate between IDCd and CDSA was 90.6% (480/530). The positivity rates on IDCd on days 1 and 2 (55.6% and 85.0%, respectively) were significantly higher than those on CDSA (19.4% and 75.6%, respectively) (P<0.001 for day 1 and P=0.02 for day 2), but the detection rates on IDCd and CDSA on day 3 were not different (89.4% vs. 82.8%, P=0.0914). On day 3, the recovery rates for non-C. difficile isolates on IDCd and CDSA were 30.2% (160/530) and 22.1% (117/530), respectively (P=0.0075). Clostridium spp. other than C. difficile were the most prevalent non-C. difficile isolates on both media. Conclusions The culture positivity rates on IDCd and CDSA were not different on day 3 but IDCd may allow for rapid and sensitive detection of C. difficile within 2 days of cultivation. PMID:24422190

  8. Plasmonically amplified fluorescence bioassay with microarray format

    NASA Astrophysics Data System (ADS)

    Gogalic, S.; Hageneder, S.; Ctortecka, C.; Bauch, M.; Khan, I.; Preininger, Claudia; Sauer, U.; Dostalek, J.

    2015-05-01

    Plasmonic amplification of fluorescence signal in bioassays with microarray detection format is reported. A crossed relief diffraction grating was designed to couple an excitation laser beam to surface plasmons at the wavelength overlapping with the absorption and emission bands of fluorophore Dy647 that was used as a label. The surface of periodically corrugated sensor chip was coated with surface plasmon-supporting gold layer and a thin SU8 polymer film carrying epoxy groups. These groups were employed for the covalent immobilization of capture antibodies at arrays of spots. The plasmonic amplification of fluorescence signal on the developed microarray chip was tested by using interleukin 8 sandwich immunoassay. The readout was performed ex situ after drying the chip by using a commercial scanner with high numerical aperture collecting lens. Obtained results reveal the enhancement of fluorescence signal by a factor of 5 when compared to a regular glass chip.

  9. Superluminescent variants of marine luciferases for bioassays.

    PubMed

    Kim, Sung Bae; Suzuki, Hideyuki; Sato, Moritoshi; Tao, Hiroaki

    2011-11-15

    In this study, a rational synthesis of superluminescent variants from marine luciferases with prolonged bioluminescence has been demonstrated. A putative active site of a model marine luciferase, Gaussia princeps Luciferase (GLuc), was assigned and modified by a site-directed mutagenesis. The potent variants were found to generate up to 10 times stronger bioluminescence, emitting red shifts of up to 33 nm with natural coelenterazine than native GLuc, rendering an efficient optical signature in bioassays. The advantageous properties were demonstrated with mammalian two-hybrid assays, single-chain probes, and metastases of murine B16 melanoma in BALB/c nude mice. The unique ideas for engineering GLuc are proved to be valid even for other marine luciferases. PMID:21951281

  10. Cell-based bioassays in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Itle, Laura J.; Zguris, Jeanna C.; Pishko, Michael V.

    2004-12-01

    The development of cell-based bioassays for high throughput drug screening or the sensing of biotoxins is contingent on the development of whole cell sensors for specific changes in intracellular conditions and the integration of those systems into sample delivery devices. Here we show the feasibility of using a 5-(and-6)-carboxy SNARF-1, acetoxymethyl ester, acetate, a fluorescent dye capable of responding to changes in intracellular pH, as a detection method for the bacterial endotoxin, lipopolysaccharide. We used photolithography to entrap cells with this dye within poly(ethylene) glyocol diacrylate hydrogels in microfluidic channels. After 18 hours of exposure to lipopolysaccharide, we were able to see visible changes in the fluorescent pattern. This work shows the feasibility of using whole cell based biosensors within microfluidic networks to detect cellular changes in response to exogenous agents.

  11. 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.

  12. Green synthesis of gold nanoparticles of different sizes and shapes using agar-agar water solution and femtosecond pulse laser irradiation

    NASA Astrophysics Data System (ADS)

    Almeida de Matos, Ricardo; da Silva Cordeiro, Thiago; Elgul Samad, Ricardo; Dias Vieira, Nilson; Coronato Courrol, Lilia

    2012-11-01

    We report a method to create gold nanoparticles of different sizes and shapes using agar-agar water solution and irradiation with light from a xenon lamp, followed by ultrashort laser pulses. No additives, such as solvents, surfactants or reducing agents, were used in the procedure. Laser irradiation (laser ablation) was important to the reduction of the nanoparticles diameter and formation of another shapes. Distilled water was used as solvent and agar-agar (hydrophilic colloid extracted from certain seaweeds) was important for the stabilization of gold nanoparticles, avoiding their agglomeration. The formation of gold nanoparticles was confirmed with ultraviolet-visible absorption and TEM microscopy. The gold nanoparticles acquired spherical, prism, and rod shapes depending on the laser parameters. Variation of laser irradiation parameters as pulse energy, irradiation time and repetition rate was assessed. The relevant mechanisms contributing for the gold nanoparticles production are discussed.

  13. 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

  14. PubChem BioAssay: 2014 update.

    PubMed

    Wang, Yanli; Suzek, Tugba; Zhang, Jian; Wang, Jiyao; He, Siqian; Cheng, Tiejun; Shoemaker, Benjamin A; Gindulyte, Asta; Bryant, Stephen H

    2014-01-01

    PubChem's BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for archiving biological tests of small molecules generated through high-throughput screening experiments, medicinal chemistry studies, chemical biology research and drug discovery programs. In addition, the BioAssay database contains data from high-throughput RNA interference screening aimed at identifying critical genes responsible for a biological process or disease condition. The mission of PubChem is to serve the community by providing free and easy access to all deposited data. To this end, PubChem BioAssay is integrated into the National Center for Biotechnology Information retrieval system, making them searchable by Entrez queries and cross-linked to other biomedical information archived at National Center for Biotechnology Information. Moreover, PubChem BioAssay provides web-based and programmatic tools allowing users to search, access and analyze bioassay test results and metadata. In this work, we provide an update for the PubChem BioAssay resource, such as information content growth, new developments supporting data integration and search, and the recently deployed PubChem Upload to streamline chemical structure and bioassay submissions. PMID:24198245

  15. PubChem BioAssay: 2014 update

    PubMed Central

    Wang, Yanli; Suzek, Tugba; Zhang, Jian; Wang, Jiyao; He, Siqian; Cheng, Tiejun; Shoemaker, Benjamin A.; Gindulyte, Asta; Bryant, Stephen H.

    2014-01-01

    PubChem’s BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for archiving biological tests of small molecules generated through high-throughput screening experiments, medicinal chemistry studies, chemical biology research and drug discovery programs. In addition, the BioAssay database contains data from high-throughput RNA interference screening aimed at identifying critical genes responsible for a biological process or disease condition. The mission of PubChem is to serve the community by providing free and easy access to all deposited data. To this end, PubChem BioAssay is integrated into the National Center for Biotechnology Information retrieval system, making them searchable by Entrez queries and cross-linked to other biomedical information archived at National Center for Biotechnology Information. Moreover, PubChem BioAssay provides web-based and programmatic tools allowing users to search, access and analyze bioassay test results and metadata. In this work, we provide an update for the PubChem BioAssay resource, such as information content growth, new developments supporting data integration and search, and the recently deployed PubChem Upload to streamline chemical structure and bioassay submissions. PMID:24198245

  16. Pulsed photothermal temperature profiling of agar tissue phantoms.

    PubMed

    Milanic, Matija; Majaron, Boris; Nelson, J Stuart

    2007-11-01

    We determine experimentally the accuracy of pulsed photothermal radiometric (PPTR) temperature depth profiling in water-based samples. We use custom tissue phantoms composed of agar gel layers separated by very thin absorbing layers. Two configurations of the acquisition system are compared, one using the customary spectral band of the InSb radiation detector (3.0-5.5 microm) and the other with a spectrally narrowed acquisition band (4.5-5.5 microm). The laser-induced temperature depth profiles are reconstructed from measured radiometric signals using a custom minimization algorithm. The results correlate very well with phantom geometry as determined by optical coherence tomography (OCT) and histology in all evaluated samples. Determination of the absorbing layer depth shows good repeatability with spatial resolution decreasing with depth. Spectral filtering improves the accuracy and resolution, especially for shallow absorption layers (~120 microm) and more complex structures (e.g., with two absorbing layers). The average full width at half maximum (FWHM) of the temperature peaks equals 23% of the layer depth. PMID:17522951

  17. Collection and control of tritium bioassay samples at Pantex

    SciTech Connect

    Fairrow, N.L.; Ivie, W.E.

    1992-01-01

    Pantex is the final assembly/disassembly point for US nuclear weapons. The Pantex internal dosimetry section monitors radiation workers once a month for tritium exposure. In order to manage collection and control of the bioassay specimens efficiently, a bar code system for collection of samples was developed and implemented to speed up the process and decrease the number of errors probable when transferring data. In the past, all the bioassay data from samples were entered manually into a computer database. Transferring the bioassay data from the liquid scintillation counter to each individual's dosimetry record required as much as two weeks of concentrated effort.

  18. Collection and control of tritium bioassay samples at Pantex

    SciTech Connect

    Fairrow, N.L.; Ivie, W.E.

    1992-12-31

    Pantex is the final assembly/disassembly point for US nuclear weapons. The Pantex internal dosimetry section monitors radiation workers once a month for tritium exposure. In order to manage collection and control of the bioassay specimens efficiently, a bar code system for collection of samples was developed and implemented to speed up the process and decrease the number of errors probable when transferring data. In the past, all the bioassay data from samples were entered manually into a computer database. Transferring the bioassay data from the liquid scintillation counter to each individual`s dosimetry record required as much as two weeks of concentrated effort.

  19. Estrogen Receptor Agonists and Antagonists in the Yeast Estrogen Bioassay.

    PubMed

    Wang, Si; Bovee, Toine F H

    2016-01-01

    Cell-based bioassays can be used to predict the eventual biological activity of a substance on a living organism. In vitro reporter gene bioassays are based on recombinant vertebrate cell lines or yeast strains and especially the latter are easy-to-handle, cheap, and fast. Moreover, yeast cells do not express estrogen, androgen, progesterone or glucocorticoid receptors, and are thus powerful tools in the development of specific reporter gene systems that are devoid of crosstalk from other hormone pathways. This chapter describes our experience with an in-house developed RIKILT yeast estrogen bioassay for testing estrogen receptor agonists and antagonists, focusing on the applicability of the latter. PMID:26585147

  20. Determining Enzyme Activity by Radial Diffusion

    ERIC Educational Resources Information Center

    Davis, Bill D.

    1977-01-01

    Discusses advantages of radial diffusion assay in determining presence of enzyme and/or rough approximation of amount of enzyme activities. Procedures are included for the preparation of starch-agar plates, and the application and determination of enzyme. Techniques using plant materials (homogenates, tissues, ungerminated embryos, and seedlings)…

  1. Physicochemical and morphological properties of plasticized poly(vinyl alcohol)-agar biodegradable films.

    PubMed

    Madera-Santana, T J; Freile-Pelegrín, Y; Azamar-Barrios, J A

    2014-08-01

    The effects of the addition of glycerol (GLY) on the physicochemical and morphological properties of poly(vinyl alcohol) (PVA)-agar films were reported. PVA-agar films were prepared by solution cast method, and the addition of GLY in PVA-agar films altered the optical properties, resulting in a decrease in opacity values and in the color difference (ΔE) of the films. Structural characterization using Fourier transformation infrared (FTIR) spectroscopy and X-ray diffraction (XRD) indicated that the presence of GLY altered the intensity of the bands (from 1200 to 800cm(-1)) and crystallinity. The characterization of the thermal properties indicated that an increase in the agar content produces a decrease in the melting temperature and augments the heat of fusion. Similar tendencies were observed in plasticized films, but at different magnification. The formulation that demonstrated the lowest mechanical properties contained 25wt.% agar, whereas the formulation that contained 75wt.% agar demonstrated a significant improvement. The water vapor transmission rate (WVTR) and surface morphology analysis demonstrated that the structure of PVA-agar films is reorganized upon GLY addition. The physicochemical properties of PVA-agar films using GLY as a plasticizer provide information for the application of this formulation as packaging material for specific food applications. PMID:24875313

  2. 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. PMID:25439904

  3. 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

  4. 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. PMID:14575957

  5. 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

  6. 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

  7. 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

  8. [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

  9. Evaporation-Driven Bioassays in Suspended Droplets.

    PubMed

    Hernandez-Perez, Ruth; Fan, Z Hugh; Garcia-Cordero, Jose L

    2016-07-19

    The microtiter plate has been an essential tool for diagnostics, high-throughput screening, and biological assays. We present an alternative platform to perform bioassays in a microplate format that exploits evaporation to drive assay reactions. Our method consists of droplets suspended on plastic pillars; reactions occur in these droplets instead of the wells. The pillars are fabricated by milling, and the rough surface created by this fabrication method pins the droplet to a constant contact line during the assay and also acts as a hydrophobic surface. Upon evaporation, natural convection arising from Marangoni currents mixes solutions in the droplet, which speeds up assay reactions, decreases assay times, and increases limits of detection. As a proof of concept we implemented two colorimetric assays to detect glucose and proteins in only 1.5 μL, without any external devices for mixing and with a digital microscope as a readout mechanism. Our platform is an ideal alternative to the microtiter plate, works with different volumes, is compatible with commercially available reagent dispensers and plate-readers, and could have broad applications in diagnostics and high-throughput screening. PMID:27331825

  10. Annotating Human P-Glycoprotein Bioassay Data

    PubMed Central

    Zdrazil, Barbara; Pinto, Marta; Vasanthanathan, Poongavanam; Williams, Antony J; Balderud, Linda Zander; Engkvist, Ola; Chichester, Christine; Hersey, Anne; Overington, John P; Ecker, Gerhard F

    2012-01-01

    Abstract Huge amounts of small compound bioactivity data have been entering the public domain as a consequence of open innovation initiatives. It is now the time to carefully analyse existing bioassay data and give it a systematic structure. Our study aims to annotate prominent in vitro assays used for the determination of bioactivities of human P-glycoprotein inhibitors and substrates as they are represented in the ChEMBL and TP-search open source databases. Furthermore, the ability of data, determined in different assays, to be combined with each other is explored. As a result of this study, it is suggested that for inhibitors of human P-glycoprotein it is possible to combine data coming from the same assay type, if the cell lines used are also identical and the fluorescent or radiolabeled substrate have overlapping binding sites. In addition, it demonstrates that there is a need for larger chemical diverse datasets that have been measured in a panel of different assays. This would certainly alleviate the search for other inter-correlations between bioactivity data yielded by different assay setups. PMID:23293680

  11. Bioassay-Directed Fractionation of Diesel and Biodiesel Emissions

    EPA Science Inventory

    Biofuels are being developed as alternatives to petroleum-derived products, but published research is contradictory regarding the mutagenic activity of such emissions relative to those from petroleum diesel. We performed bioassay-directed fractionation and analyzed the polycyclic...

  12. Bioassay Phantoms Using Medical Images and Computer Aided Manufacturing

    SciTech Connect

    Dr. X. Geroge Xu

    2011-01-28

    A radiation bioassay program relies on a set of standard human phantoms to calibrate and assess radioactivity levels inside a human body for radiation protection and nuclear medicine imaging purposes. However, the methodologies in the development and application of anthropomorphic phantoms, both physical and computational, had mostly remained the same for the past 40 years. We herein propose a 3-year research project to develop medical image-based physical and computational phantoms specifically for radiation bioassay applications involving internally deposited radionuclides. The broad, long-term objective of this research was to set the foundation for a systematic paradigm shift away from the anatomically crude phantoms in existence today to realistic and ultimately individual-specific bioassay methodologies. This long-term objective is expected to impact all areas of radiation bioassay involving nuclear power plants, U.S. DOE laboratories, and nuclear medicine clinics.

  13. A CONTROLLED BIOASSAY SYSTEM FOR MEASURING TOXICITY OF HEAVY METALS

    EPA Science Inventory

    Biological availability of metal micronutrients and metal toxicity are believed to be dependent on metal oxidation state, complexation, and solubility as well as the physicochemical characteristics of the aqueous phase. Basic design criteria for fish bioassays which are capable o...

  14. A wind tunnel bioassay system for screening mosquito repellents.

    PubMed

    Sharpington, P J; Healy, T P; Copland, M J

    2000-09-01

    A wind tunnel bioassay system to screen mosquito repellents is described. A wind tunnel is utilized to exploit the upwind flight response of host-seeking mosquitoes. Mosquitoes within the wind tunnel are activated with human breath, fly upwind, and land on heated chick skins. This behavioral sequence results in a consistently high percentage of the test population approaching repellent or control stimuli. The bioassay system is calibrated with diethyl methylbenzamide against Aedes aegypti and demonstrates a reproducible dose-response relationship. The persistence of diethyl methyl benzamide after a 1-h period is also recorded. The design of the bioassay system permits simultaneous, independent testing of 3 candidate repellents. The wind tunnel bioassay system is compared to other techniques for evaluating mosquito repellents. PMID:11081652

  15. Comparison of laboratory batch and flow-through microcosm bioassays.

    PubMed

    Clément, Bernard J P; Delhaye, Hélène L; Triffault-Bouchet, Gaëlle G

    2014-10-01

    Since 1997, we have been developing a protocol for ecotoxicological bioassays in 2-L laboratory microcosms and have applied it to the study of various pollutants and ecotoxicological risk assessment scenarios in the area of urban facilities and transport infrastructures. The effects on five different organisms (micro-algae, duckweeds, daphnids, amphipods, chironomids) are assessed using biological responses such as growth, emergence (chironomids), reproduction (daphnids) and survival, with a duration of exposure of 3 weeks. This bioassay has mainly been used as a batch bioassay, i.e., the water was not renewed during the test. A flow-through microcosm bioassay has been developed recently, with the assumption that conditions for the biota should be improved, variability reduced, and the range of exposure patterns enlarged (e.g., the possibility of maintaining constant exposure in the water column). This paper compares the results obtained in batch and flow-through microcosm bioassays, using cadmium as a model toxicant. As expected, the stabilization of physico-chemical parameters, increased organism fitness and reduced variability were observed in the flow-through microcosm bioassay. PMID:25086825

  16. Acarine attractants: Chemoreception, bioassay, chemistry and control.

    PubMed

    Carr, Ann L; Roe, Michael

    2016-07-01

    The Acari are of significant economic importance in crop production and human and animal health. Acaricides are essential for the control of these pests, but at the same time, the number of available pesticides is limited, especially for applications in animal production. The Acari consist of two major groups, the mites that demonstrate a wide variety of life strategies, i.e., herbivory, predation and ectoparasitism, and ticks which have evolved obligatory hematophagy. The major sites of chemoreception in the acarines are the chelicerae, palps and tarsi on the forelegs. A unifying name, the "foretarsal sensory organ" (FSO), is proposed for the first time in this review for the sensory site on the forelegs of all acarines. The FSO has multiple sensory functions including olfaction, gustation, and heat detection. Preliminary transcriptomic data in ticks suggest that chemoreception in the FSO is achieved by a different mechanism from insects. There are a variety of laboratory and field bioassay methods that have been developed for the identification and characterization of attractants but minimal techniques for electrophysiology studies. Over the past three to four decades, significant progress has been made in the chemistry and analysis of function for acarine attractants in mites and ticks. In mites, attractants include aggregation, immature female, female sex and alarm pheromones; in ticks, the attraction-aggregation-attachment, assembly and sex pheromones; in mites and ticks host kairomones and plant allomones; and in mites, fungal allomones. There are still large gaps in our knowledge of chemical communication in the acarines compared to insects, especially relative to acarine pheromones, and more so for mites than ticks. However, the use of lure-and-kill and lure-enhanced biocontrol strategies has been investigated for tick and mite control, respectively, with significant environmental advantages which warrant further study. PMID:27265828

  17. Detection of Agar, by Analysis of Sugar Markers, Associated with Bacillus Anthracis Spores, After Culture

    SciTech Connect

    Wunschel, David S.; Colburn, Heather A.; Fox, Alvin; Fox, Karen F.; Harley, William M.; Wahl, Jon H.; Wahl, Karen L.

    2008-08-01

    Detection of small quantities of agar associated with spores of Bacillus anthracis could provide key information regarding its source or growth characteristics. Agar, widely used in growth of bacteria on solid surfaces, consists primarily of repeating polysaccharide units of 3,6-anhydro-L-galactose (AGal) and galactose (Gal) with sulfated and O-methylated galactoses present as minor constituents. Two variants of the alditol acetate procedure were evaluated for detection of potential agar markers associated with spores. The first method employed a reductive hydrolysis step, to stabilize labile anhydrogalactose, by converting to anhydrogalactitol. The second eliminated the reductive hydrolysis step simplifying the procedure. Anhydrogalactitol, derived from agar, was detected using both derivatization methods followed by gas chromatography-mass spectrometry (GC-MS) analysis. However, challenges with artefactual background (reductive hydrolysis) or marker destruction (hydrolysis) lead to the search for alternative sugar markers. A minor agar component, 6-O-methyl galactose (6-O-M gal), was readily detected in agar-grown but not broth-grown bacteria. Detection was optimized by the use of gas chromatography-tandem mass spectrometry (GC-MS-MS). With appropriate choice of sugar marker and analytical procedure, detection of sugar markers for agar has considerable potential in microbial forensics.

  18. Factors Affecting Selectivity of Brilliant Green-Phenol Red Agar for Salmonellae

    PubMed Central

    Moats, W. A.; Kinner, J. A.

    1974-01-01

    Commercial brilliant green (BG)-sulfa agar was found to be nonselective toward a test series of Enterobacteriaceae. Various formulations of BG were prepared by using Trypticase soy agar (BBL) as a base. Results were more reproducible when BG dye was added after sterilization than before. Sulfonamides improved selectivity as compared with brilliant green alone. Sulfanilamide (SN) was slightly more selective for salmonellae than other sulfonamides tested. Bile salts and sodium dodecyl sulfate markedly reduced the toxicity of BG to all the test bacteria. Enterobacter strains were most difficult to inhibit. A combination of 5 mg of BG and 1 g of SN/liter prevented growth of Proteus mirabilis and Escherichia coli and retarded growth of Enterobacter strains. The BG-SN agars were superior in selectivity to a series of commercial agars tested, and numbers of salmonellae recovered on BG-SN agar and Trypticase soy agar (BBL) were the same. Brilliant green agars with various degrees of selectivity are described. PMID:4589120

  19. 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.

  20. Effect of heat treatment on the performance of tryptose-sulfite-cycloserine agar for enumeration of Clostridium perfringens.

    PubMed

    Brodsky, M H; Ciebin, B W

    1979-05-01

    Dissolving dehydrated tryptose-sulfite-cycloserine agar by only boiling or microwaving was found to inhibit Clostridium perfringens colony development in pour plates when compared with C. perfringens recovery in tryptose-sulfite-cycloserine agar prepared by autoclaving. PMID:225988

  1. A novel laboratory screening bioassay for crop seedling allelopathy.

    PubMed

    Belz, Regina G; Hurle, Karl

    2004-01-01

    Crops that control weeds by root exudation of allelochemicals are receiving increased attention, and there are efforts to breed allelopathic cultivars in several crops. The genetic improvement of allelopathic traits is based upon parental germ plasm with high allelopathic activity. Identification of allelopathic germplasm is done in laboratory screening bioassays, but experimental protocols are limited. We developed a fast and reliable laboratory screening bioassay for grain crops that includes dose-response considerations as an integral part of the experimental design. The bioassay was conducted in hydroponic culture, and a range of experiments with 2-(3H)-benzoxazolinone (BOA), an allelochemical of several grain crops, was carried out to define the basic protocol. Because of its sensitivity to BOA, Sinapis alba L. was selected as the receiver species. BOA affected growth (fresh weight and length of shoot and root), enzyme activities (ascorbate peroxidase, catalase, glutathione S-transferase, peroxidase, phenylalanine ammonia-lyase), and chlorophyll fluorescence, whereby root length was the most reliable response parameter. BOA sensitivity was dependent on nutrients for all parameters measured, and, thus, no nutrients were added. A set of experiments with Secale cereale L. and Triticum aestivum L. as donor species was carried out to optimize the protocol. Light and pH were eliminated as primary causes for the observed inhibition. The proposed bioassay has several methodological advantages over current bioassays. PMID:15074665

  2. Soil bioassays and the {sup 129}I problem

    SciTech Connect

    Sheppard, S.C.

    1995-12-31

    Iodine-129 is a very long-lived radionuclide associated with spent nuclear fuel. Because {sup 129}I has a 10{sup 7}-year half-life, is very mobile in the environment and is a biologically essential element, it is the most limiting radionuclide affecting disposal of spent fuel. Traditionally, the potential impacts of {sup 129}I have been estimated for human receptors, with the implicit assumption that all other organisms are less at risk. Risk is the operative word, the objective for protection of humans is to protect individuals, whereas the objective for other biota is usually to protect populations. Here, {sup 129}I poses an interesting problem: the half-life is so long it is barely radioactive. Thus, the chemical toxicity may be more limiting than the radiological impact. A series of soil bioassays were employed, including a life-cycle plant (Brassica rapa) bioassay, a modified earthworm survival bioassay, a microarthropod colonization/survival bioassay, and a series of more common soil and aquatic bioassays. Chemical toxicity was indicated at soil concentrations as low as 5 mg kg{sup {minus}1}. At these levels, radiological impact on non-human biota would not be expected, and therefore the chemical toxicity effects are more critical. However, human food-chain model estimates show these levels, as pure {sup 129}I, would be unacceptable for human radiological exposure, so that for {sup 129}I, protection of the human environment should also be protective of non-human biota.

  3. 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.

  4. [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

  5. [Investigation on pattern and methods of quality control for Chinese materia medica based on dao-di herbs and bioassay - bioassay for Coptis chinensis].

    PubMed

    Yan, Dan; Xiao, Xiao-he

    2011-05-01

    Establishment of bioassay methods is the technical issues to be faced with in the bioassay of Chinese materia medica. Taking the bioassay of Coptis chinensis Franch. as an example, the establishment process and application of the bioassay methods (including bio-potency and bio-activity fingerprint) were explained from the aspects of methodology, principle of selection, experimental design, method confirmation and data analysis. The common technologies were extracted and formed with the above aspects, so as to provide technical support for constructing pattern and method of the quality control for Chinese materia medica based on the dao-di herbs and bioassay. PMID:21800546

  6. 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

  7. 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.

  8. MRI Phantoms – Are There Alternatives to Agar?

    PubMed Central

    Hellerbach, Alexandra; Schuster, Verena; Jansen, Andreas; Sommer, Jens

    2013-01-01

    The suitability of different gelling agents as MRI phantoms was evaluated in terms of homogeneity, gel stability and reproducibility. Time and effort for preparation were also taken into account. The relaxation times of various gel compositions were estimated. Carbomer-980 and Carbopol-974P were determined to be promising novel phantom materials. These gelling agents are readily available, inexpensive and easy to handle given that thermal treatment is not required. Furthermore, the viscoelasticity of their polymer network is pH-dependent. With such characteristics, it was even possible to embed sensitive objects and retrieve them after testing. This was demonstrated with a fiber phantom for Diffusion Weighted MRI applications. Since Carbomer-980 and Carbopol-974P are non-hazardous, they are also suitable for multimodal setups (e.g., MRI as well as ultrasonic imaging). PMID:23940563

  9. 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.

  10. Rapid detection of Clostridium perfringens: comparison of lactose sulfite broth with tryptose-sulfite-cycloserine agar.

    PubMed

    Neut, C; Pathak, J; Romond, C; Beerens, H

    1985-01-01

    The lactose sulfite (LS) medium recommended for the detection and identification of Clostridium perfringens in foods was compared with a reference method using tryptose-sulfite-cycloserine (TSC) agar for the enumeration of this organism in a variety of foods and food ingredients. C. perfringens was detected and enumerated in 17 of the 54 samples examined with LS broth, but its presence could be confirmed in only 9 of the samples with TSC agar. In only 2 instances, C. perfringens was detected on TSC agar but not in LS broth. A positive response (FeS + and gas +) in LS broth incubated at 46 degrees C always corresponded to the presence of C. perfringens; whereas the black colonies formed on TSC agar incubated at 37 degrees C were frequently found to be Clostridium species other than C. perfringens. Thus, because of its highly selective nature, LS broth was superior to TSC agar for enumerating and confirming the small numbers of C. perfringens that were present in a majority of the samples. This was especially true when other clostridia were also present. Besides its greater selectivity and sensitivity, LS broth had the additional advantages of requiring less work and giving confirmed results within 24-48 h compared with 3 days for the TSC agar method. PMID:2865247

  11. [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. PMID:26638483

  12. Growth of coagulase-negative staphylococci on colistin-nalidixic acid agar and susceptibility to polymyxins.

    PubMed

    Fung, J C; McKinley, G; Tyburski, M B; Berman, M; Goldstein, J

    1984-05-01

    Colistin-nalidixic acid agar, although recently recommended as a replacement for blood agar for primary plating of urine specimens ( Fung et al., J. Clin. Microbiol. 16:632-636, 1982), has also been reported to suppress the growth of some strains of staphylococci that are susceptible to colistin (polymyxin E). The susceptibility of 11 species of staphylococci to polymyxins was determined, and the ability of these species to grow on colistin-nalidixic acid agar was examined. Although the MICs for most of the strains tested were 8 micrograms/ml or less, only a few coagulase-negative staphylococci grew on or were inhibited by colistin-nalidixic acid agar. This descrepancy was explained by the antagonistic effects that medium components, such as physiological concentrations of magnesium and calcium and 5% sheep blood, had on the activity of polymyxin. Colistin-nalidixic acid agar is still recommended for routine urine processing; however, the poor growth of 13% of the Staphylococcus saprophyticus strains tested suggests that blood agar should be included in the primary plating battery of urine specimens obtained from female outpatients. PMID:6330170

  13. Antimicrobial Activity and Brine Shrimp Lethality Bioassay of the Leaves Extract of Dillenia indica Linn

    PubMed Central

    Apu, AS; Muhit, MA; Tareq, SM; Pathan, AH; Jamaluddin, ATM; Ahmed, M

    2010-01-01

    The crude methanolic extract of Dillenia indica Linn. (Dilleniaceae) leaves has been investigated for the evaluation of antimicrobial and cytotoxic activities. Organic solvent (n-hexane, carbon tetrachloride and chloroform) fractions of methanolic extract and methanolic fraction (aqueous) were screened for their antimicrobial activity by disc diffusion method. Besides, the fractions were screened for cytotoxic activity using brine shrimp (Artemia salina) lethality bioassay. Among the four fractions tested, n-hexane, carbon tetrachloride, and chloroform fractions showed moderate antibacterial and antifungal activity compared to standard antibiotic, kanamycin. The average zone of inhibition was ranged from 6 to 8 mm at a concentration of 400 µg/disc. But the aqueous fraction was found to be insensitive to microbial growth. Compared to vincristine sulfate (with LC50 of 0.52 µg/ ml), n-hexane and chloroform fractions demonstrated a significant cytotoxic activity (having LC50 of 1.94 µg/ml and 2.13 µg/ml, respectively). The LC50 values of the carbon tetrachloride and aqueous fraction were 4.46 µg/ml and 5.13 µg/ ml, respectively. The study confirms the moderate antimicrobial and potent cytotoxic activities of Dillenia indica leaves extract and therefore demands the isolation of active principles and thorough bioassay. PMID:21331191

  14. Antimicrobial Activity and Brine Shrimp Lethality Bioassay of the Leaves Extract of Dillenia indica Linn.

    PubMed

    Apu, As; Muhit, Ma; Tareq, Sm; Pathan, Ah; Jamaluddin, Atm; Ahmed, M

    2010-01-01

    The crude methanolic extract of Dillenia indica Linn. (Dilleniaceae) leaves has been investigated for the evaluation of antimicrobial and cytotoxic activities. Organic solvent (n-hexane, carbon tetrachloride and chloroform) fractions of methanolic extract and methanolic fraction (aqueous) were screened for their antimicrobial activity by disc diffusion method. Besides, the fractions were screened for cytotoxic activity using brine shrimp (Artemia salina) lethality bioassay. Among the four fractions tested, n-hexane, carbon tetrachloride, and chloroform fractions showed moderate antibacterial and antifungal activity compared to standard antibiotic, kanamycin. The average zone of inhibition was ranged from 6 to 8 mm at a concentration of 400 µg/disc. But the aqueous fraction was found to be insensitive to microbial growth. Compared to vincristine sulfate (with LC(50) of 0.52 µg/ ml), n-hexane and chloroform fractions demonstrated a significant cytotoxic activity (having LC(50) of 1.94 µg/ml and 2.13 µg/ml, respectively). The LC(50) values of the carbon tetrachloride and aqueous fraction were 4.46 µg/ml and 5.13 µg/ ml, respectively. The study confirms the moderate antimicrobial and potent cytotoxic activities of Dillenia indica leaves extract and therefore demands the isolation of active principles and thorough bioassay. PMID:21331191

  15. Environmental effects of dredging. A chronic sublethal sediment bioassay with the marine polychaete nereis (Neanthes) arenaceodentata

    SciTech Connect

    Dillon, T.M.; Moore, D.W.; Bridges, T.S.

    1995-01-01

    This note provides a general overview of a new 28-day chronic sublethal sediment bioassay designed for the regulatory evaluation of dredged material. The bioassay uses survival and growth rate endpoints with the polychaete Nereis (Neanthes) arenaceodentata. The primary technical reference for this new bioassay is Dillon, Moore, and Reish (in press), upon which this overview is based. Sediment bioassays are used to assess the aggregate toxicity of sediment associated anthropogenic chemicals. Historically, these bioassays have measured survival of highly sensitive species following acute exposures (10 days). A new generation of sediment bioassays is being developed in which the subtle, sublethal response of test species is measured following chronic sediment exposures (Dillon 1993).

  16. Carbon-14 Bioassay for Decommissioning of Hanford Reactors

    SciTech Connect

    Carbaugh, Eugene H.; Watson, David J.

    2012-05-01

    The old production reactors at the US Department of Energy Hanford Site used large graphite piles as the moderator. As part of long-term decommissioning plans, the potential need for 14C radiobioassay of workers was identified. Technical issues associated with 14C bioassay and worker monitoring were investigated, including anticipated graphite characterization, potential intake scenarios, and the bioassay capabilities that may be required to support the decommissioning of the graphite piles. A combination of urine and feces sampling would likely be required for the absorption type S 14C anticipated to be encountered. However the concentrations in the graphite piles appear to be sufficiently low that dosimetrically significant intakes of 14C are not credible, thus rendering moot the need for such bioassay.

  17. Method comparison for 241Am emergency urine bioassay.

    PubMed

    Li, Chunsheng; Sadi, Baki; Benkhedda, Karima; St-Amant, Nadereh; Moodie, Gerry; Ko, Raymond; Dinardo, Anthony; Kramer, Gary

    2010-10-01

    241Am is one of the high-risk radionuclides that might be used in a terrorist attack. 241Am in urine bioassay can identify the contaminated individuals who need immediate medical intervention and decontamination. This paper compares three methods for the measurement of 241Am in urine, namely liquid scintillation counting (LSC), inductively coupled plasma mass spectrometry (ICP-MS) and gamma spectrometry (GS), at two levels, 20 and 2 Bq l(-1). All three methods satisfied the ANSI N13.30 radio-bioassay criteria for accuracy and repeatability. ICP-MS offered the best sensitivity and fastest sample turnaround; however, the ICP-MS system used in this work may not be available in many bioassay laboratories. LSC and GS are more commonly available instruments. GS requires minimal or no sample preparation, which makes it a good candidate method. Moreover, the sample throughput can be significantly improved if the GS and LSC methods are automated. PMID:20573683

  18. Internal dosimetry performing dose assessments via bioassay measurements

    SciTech Connect

    Bailey, K.M.

    1993-05-11

    The Internal Dosimetry Department at the Y-12 Plant maintains a state-of-the-art bioassay program managed under the guidance and regulations of the Department of Energy. The two major bioassay techniques currently used at Y-12 are the in vitro (urinalysis) and in vivo (lung counting) programs. Fecal analysis (as part of the in vitro program) is another alternative; however, since both urine and fecal analysis provide essentially the same capabilities for detecting exposures to uranium, the urinalysis is the main choice primarily for aesthetic reasons. The bioassay frequency is based on meeting NCRP 87 objectives which are to monitor the accumulation of radioactive material in exposed individuals, and to ensure that significant depositions are detected.

  19. The effect of pesticide residue on caged mosquito bioassays.

    PubMed

    Barber, J A S; Greer, Mike; Coughlin, Jamie

    2006-09-01

    Wind tunnel experiments showed that secondary pickup of insecticide residue by mosquitoes in cage bioassays had a significant effect on mortality. Cage bioassays using adult Ochlerotatus taeniorhynchus (Wiedemann) investigated the effect of exposure time to a contaminated surface. Cages were dosed in a wind tunnel using the LC50 for naled (0.124 mg a.i./ml) and an LC25 (0.0772 mg a.i./ml) for naled. Half of the bioassay mosquitoes were moved directly into clean cages with the other half remaining in the sprayed, hence contaminated, cage. Treatment mortality was assessed at 8, 15, 30, 60, 120, 240, and 1,440 min postapplication. Cage contamination had a significant effect on mosquito mortality for both the LC25 and LC50 between 15 and 30 min postapplication. PMID:17067048

  20. Bioassay-directed chemical analysis in environmental research

    SciTech Connect

    Schuetzle, D.; Lewtas, J.

    1986-01-01

    The use of short-term bioassay tests in conjunction with analytical measurements, constitute a powerful tool for identifying important environmental contaminants. The authors have coined the terminology bioassay directed chemical analysis to best describe this marriage of analytical chemistry and biology. The objective of this methodology is to identify key compounds in various types of air-pollutant samples. Once that task is completed, studies on metabolism, sources, environmental exposure and atmospheric chemistry can be undertaken. The principles and methodologies for bioassay directed chemical analysis are presented and illustrated in this paper. Most of this work has been directed toward the characterization of ambient air and diesel particulates, which are used as examples in this report to illustrate the analytical logic used for identifying the bio-active components of complex mixtures.

  1. Do we really need in-situ bioassays?

    SciTech Connect

    Salazar, M.H.; Salazar, S.M.

    1995-12-31

    In-situ bioassays are needed to validate the results from laboratory testing and to understand biological interactions. Standard laboratory protocols provide reproducible test results, and the precision of those tests can be mathematically defined. Significant correlations between toxic substances and levels of response (bioaccumulation and bioeffects) have also been demonstrated with natural field populations and suggest that the laboratory results can accurately predict field responses. An equal number of studies have shown a lack of correlation between laboratory bioassay results and responses of natural field populations. The best way to validate laboratory results is with manipulative field testing; i.e., in-situ bioassays with caged organisms. Bioaccumulation in transplanted bivalves has probably been the most frequently used form of an in-situ bioassay. The authors have refined those methods to include synoptic measurements of bioaccumulation and growth. Growth provides an easily-measured bioeffects endpoint and a means of calibrating bioaccumulation. Emphasis has been on minimizing the size range of test animals, repetitive measurements of individuals and standardization of test protocols for a variety of applications. They are now attempting to standardize criteria for accepting and interpreting data in the same way that laboratory bioassays have been standardized. Others have developed methods for in-situ bioassays using eggs, larvae, unicellular organisms, crustaceans, benthic invertebrates, bivalves, and fish. In the final analysis, the in-situ approach could be considered as an exposure system where any clinical measurements are possible. The most powerful approach would be to use the same species in laboratory and field experiments with the same endpoints.

  2. Selective agars for the isolation of Streptococcus iniae from Japanese flounder, Paralichthys olivaceus, and its cultural environment.

    PubMed

    Nguyen, H T; Kanai, K

    1999-05-01

    Two kinds of selective agar were developed for the isolation of Streptococcus iniae, the causal agent of streptococcosis, from Japanese flounder (Paralichthys olivaceus) and from culture tanks in flounder farms. The selective agars were heart infusion agar with added thallium acetate and oxlinic acid (TAOA), and colistin sulphate and oxolinic acid (CSOA). For samples containing various bacterial flora, selective agars were supplemented with defibrinated horse blood in order to distinguish beta-haemolytic colonies of Strep. iniae. Streptococcus iniae was quantitatively isolated from the brain and kidney of diseased flounders in pure culture. Two-thirds of isolates picked up from selective blood agars inoculated with intestinal samples were identified as Strep. iniae. The bacterial colony numbers of deposits and water from culture tanks on selective blood agars were about 10-10(5) times smaller than those on control heart infusion agar; Strep. iniae was isolated from few deposit and water samples. PMID:10347871

  3. Rotationally Induced Hydrodynamics: Fundamentals and Applications to High-Speed Bioassays

    NASA Astrophysics Data System (ADS)

    Wang, Gufeng; Driskell, Jeremy D.; Hill, April A.; Dufek, Eric J.; Lipert, Robert J.; Porter, Marc D.

    2010-07-01

    Bioassays are indispensable tools in areas ranging from fundamental life science research to clinical practice. Improving assay speed and levels of detection will have a profound impact in all of these areas. We recently developed a rapid, sensitive format for immunosorbent assays that expedites antigen mass transport by rotating the capture substrate. This review outlines the theoretical foundation of rotationally induced hydrodynamics and its application in heterogeneous assays. We describe a general solution that solves the rates of immunoreactions on rotating capture substrates, taking into account both diffusion and the rate of reaction between antibody and antigen. The general solution applies to a wide range of rotation rates, including mass transport-limited to reaction rate-limited assays, and is validated experimentally. We discuss several applications that demonstrate how immunoassays can be tailored to increase speed as well as lower the limit of detection of viral particles, pathogens, toxins, and proteins.

  4. Mass production of spores of lactic acid-producing Rhizopus oryzae NBRC 5384 on agar plate.

    PubMed

    Yamane, Tsuneo; Tanaka, Ryosuke

    2013-01-01

    Mass production of sporangiospores (spores) of Rhizopus oryzae NBRC 5384 (identical to NRRL 395 and ATCC 9363) on potato-dextrose-agar medium was studied aiming at starting its L(+)-lactic acid fermentation directly from spore inoculation. Various parameters including harvest time, sowed spore density, size of agar plate, height of air space, and incubation mode of plate (agar-on-bottom or agar-on-top) were studied. Ordinarily used shallow Petri dishes were found out to be unsuitable for the full growth of R. oryzae sporangiophores. In a very wide range of the sowed spore density, the smaller it was, the greater the number of the harvested spores was. It was also interesting to find out that R. oryzae grown downward vertically with a deep air space in an agar-on-top mode gave larger amount of spores than in an agar-on-bottom mode at 30°C for 7-day cultivation. Scale-up of the agar plate culture from 26.4 to 292 cm(2) was studied, resulting in the proportional relationship between the number of the harvested spores/plate and the plate area in the deep Petri dishes. The number of plates of 50 cm in diameter needed for 100 m(3) industrial submerged fermentation started directly from 2 × 10(5) spores/mL inoculum size was estimated as about 6, from which it was inferred that such a fermentation would be feasible. Designing a 50 cm plate and a method of spreading and collecting the spores were suggested. Bioprocess technological significance of the "full-scale industrial submerged fermentation started directly from spore inoculation omitting pre-culture" has been discussed. PMID:23658025

  5. Evaluation of CP Chromo Select Agar for the enumeration of Clostridium perfringens from water.

    PubMed

    Manafi, Mammad; Waldherr, Kerstin; Kundi, Michael

    2013-10-01

    The European Directive on drinking water quality has included mCP agar as the reference method for recovering Clostridium perfringens from drinking waters. In the present study, three media (mCP, TSCF and CP Chromo Select Agar) were evaluated for recovery of C. perfringens in different surface water samples. Out of 139 water samples, using a membrane filtration technique, 131 samples (94.2%) were found to be presumptively positive for C. perfringens in at least one of the culture media. Green colored colonies on CP Chromo Select Agar (CCP agar) were counted as presumptive C. perfringens isolates. Out of 483 green colonies on CCP agar, 96.3% (465 strains, indole negative) were identified as C. perfringens, and 15 strains (3.1%) were indole positive and were identified as Clostridium sordellii, Clostridium bifermentans or Clostridium tetani. Only 3 strains (0.6%) gave false positive results and were identified as Clostridium fallax, Clostridium botulinum, and Clostridium tertium. Variance analysis of the data obtained shows statistically no significant differences in the counts obtained between media employed in this work. The mCP method is very onerous for routine screening and bacterial colonies could not be used for further biochemical testing. The colonies on CCP and TSCF were easy to count and subculture for confirmation tests. TSCF detects sulfite-reducing clostridia, including species other than C. perfringens, and in some cases excessive blackening of the agar frustrated counting of the colonies. If the contamination was too high, TSCF did not consistently produce black colonies and as a consequence, the colonies were white and gave false negative results. On the other hand, the identification of typical and atypical colonies isolated from all media demonstrated that CCP agar was the most useful medium for C. perfringens recovery in water samples. PMID:23816139

  6. 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

  7. A statistical treatment of bioassay pour fractions

    NASA Astrophysics Data System (ADS)

    Barengoltz, Jack; Hughes, David

    A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course

  8. A statistical treatment of bioassay pour fractions

    NASA Astrophysics Data System (ADS)

    Barengoltz, Jack; Hughes, David

    A bioassay is a method for estimating the number of bacterial spores on a spacecraft surface for the purpose of demonstrating compliance with planetary protection (PP) requirements (Ref. 1). The details of the process may be seen in the appropriate PP document (e.g., for NASA, Ref. 2). In general, the surface is mechanically sampled with a damp sterile swab or wipe. The completion of the process is colony formation in a growth medium in a plate (Petri dish); the colonies are counted. Consider a set of samples from randomly selected, known areas of one spacecraft surface, for simplicity. One may calculate the mean and standard deviation of the bioburden density, which is the ratio of counts to area sampled. The standard deviation represents an estimate of the variation from place to place of the true bioburden density commingled with the precision of the individual sample counts. The accuracy of individual sample results depends on the equipment used, the collection method, and the culturing method. One aspect that greatly influences the result is the pour fraction, which is the quantity of fluid added to the plates divided by the total fluid used in extracting spores from the sampling equipment. In an analysis of a single sample’s counts due to the pour fraction, one seeks to answer the question: What is the probability that if a certain number of spores are counted with a known pour fraction, that there are an additional number of spores in the part of the rinse not poured. This is given for specific values by the binomial distribution density, where detection (of culturable spores) is success and the probability of success is the pour fraction. A special summation over the binomial distribution, equivalent to adding for all possible values of the true total number of spores, is performed. This distribution when normalized will almost yield the desired quantity. It is the probability that the additional number of spores does not exceed a certain value. Of course

  9. HIGHLY SENSITIVE BIOASSAYS FOR EVALUATING AIRBORNE MUTAGENS INDOORS

    EPA Science Inventory

    The standard mutagenicity bioassays that are readily applied to the valuation of outdoor air samples collected by high volume samplers are not efficiently sensitive to measure the mutagenicity of low volume air samples collected indoors. wo microsuspension mutation assays using v...

  10. Filtration effects due to bioassay cage design and screen type

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of bioassay cages in the efficacy assessment of specific compounds, application techniques and technologies is a common practice. There are a number of cage designs being used that range across a variety of cage shapes and sizes and mesh types. The objective of this work was to examine a r...

  11. STRESS ETHYLENE: A BIOASSAY FOR RHIZOSPHERE-APPLIED PHYTOTOXICANTS

    EPA Science Inventory

    A bioassay for rhizosphere-applied phytotoxicants was developed and evaluated with a broad range of chemicals. Test substances were applied to the rhizosphere of whole, intact bush bean plants (Phaseolus vulgaris L. cv. Bush Blue Lake 290) grown in a solid support medium and the ...

  12. Assessment of acrylamide toxicity using a battery of standardised bioassays.

    PubMed

    Zovko, Mira; Vidaković-Cifrek, Željka; Cvetković, Želimira; Bošnir, Jasna; Šikić, Sandra

    2015-12-01

    Acrylamide is a monomer widely used as an intermediate in the production of organic chemicals, e.g. polyacrylamides (PAMs). Since PAMs are low cost chemicals with applications in various industries and waste- and drinking water treatment, a certain amount of non-polymerised acrylamide is expected to end up in waterways. PAMs are non-toxic but acrylamide induces neurotoxic effects in humans and genotoxic, reproductive, and carcinogenic effects in laboratory animals. In order to evaluate the effect of acrylamide on freshwater organisms, bioassays were conducted on four species: algae Desmodesmus subspicatus and Pseudokirchneriella subcapitata, duckweed Lemna minor and water flea Daphnia magna according to ISO (International Organization for Standardisation) standardised methods. This approach ensures the evaluation of acrylamide toxicity on organisms with different levels of organisation and the comparability of results, and it examines the value of using a battery of low-cost standardised bioassays in the monitoring of pollution and contamination of aquatic ecosystems. These results showed that EC50 values were lower for Desmodesmus subspicatus and Pseudokirchneriella subcapitata than for Daphnia magna and Lemna minor, which suggests an increased sensitivity of algae to acrylamide. According to the toxic unit approach, the values estimated by the Lemna minor and Daphnia magna bioassays, classify acrylamide as slightly toxic (TU=0-1; Class 1). The results obtained from algal bioassays (Desmodesmus subspicatus and Pseudokirchneriella subcapitata) revealed the toxic effect of acrylamide (TU=1-10; Class 2) on these organisms. PMID:26751864

  13. Artificial diets for life tables bioassays of TPB in Mississippi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two artificial diets for mass rearing and bioassay of the tarnished plant bug, (TPB), Lygus lineolaris Palisot de Beauvois, (Hemiptera: Miridae) were modified and developed, respectively. The first diet is a modification of a semisolid artificial diet (NI diet), which permits large scale rearing of ...

  14. INFLUENCE OF SEDIMENT EXTRACT FRACTIONATION METHODS ON BIOASSAY RESULTS

    EPA Science Inventory

    Four bioassays [Microtax(tm), Mutatox(tm), sister chromatid exchange (SCE), and metabolic cooperation] were used to analyze marine sediment extracts fractionated by two different methods: silica gel column chromatography and acid-base fractionation. esults indicated that a sedime...

  15. Statistical considerations in the analysis of data from replicated bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Multiple-dose bioassay is generally the preferred method for characterizing virulence of insect pathogens. Linear regression of probit mortality on log dose enables estimation of LD50/LC50 and slope, the latter having substantial effect on LD90/95s (doses of considerable interest in pest management)...

  16. Soil bioassays as tools for sludge compost quality assessment

    SciTech Connect

    Domene, Xavier; Sola, Laura; Ramirez, Wilson; Alcaniz, Josep M.; Andres, Pilar

    2011-03-15

    Composting is a waste management technology that is becoming more widespread as a response to the increasing production of sewage sludge and the pressure for its reuse in soil. In this study, different bioassays (plant germination, earthworm survival, biomass and reproduction, and collembolan survival and reproduction) were assessed for their usefulness in the compost quality assessment. Compost samples, from two different composting plants, were taken along the composting process, which were characterized and submitted to bioassays (plant germination and collembolan and earthworm performance). Results from our study indicate that the noxious effects of some of the compost samples observed in bioassays are related to the low organic matter stability of composts and the enhanced release of decomposition endproducts, with the exception of earthworms, which are favored. Plant germination and collembolan reproduction inhibition was generally associated with uncomposted sludge, while earthworm total biomass and reproduction were enhanced by these materials. On the other hand, earthworm and collembolan survival were unaffected by the degree of composting of the wastes. However, this pattern was clear in one of the composting procedures assessed, but less in the other, where the release of decomposition endproducts was lower due to its higher stability, indicating the sensitivity and usefulness of bioassays for the quality assessment of composts.

  17. US Army Radiological Bioassay and Dosimetry: The RBD software package

    SciTech Connect

    Eckerman, K. F.; Ward, R. C.; Maddox, L. B.

    1993-01-01

    The RBD (Radiological Bioassay and Dosimetry) software package was developed for the U. S. Army Material Command, Arlington, Virginia, to demonstrate compliance with the radiation protection guidance 10 CFR Part 20 (ref. 1). Designed to be run interactively on an IBM-compatible personal computer, RBD consists of a data base module to manage bioassay data and a computational module that incorporates algorithms for estimating radionuclide intake from either acute or chronic exposures based on measurement of the worker's rate of excretion of the radionuclide or the retained activity in the body. In estimating the intake,RBD uses a separate file for each radionuclide containing parametric representations of the retention and excretion functions. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent. For a given nuclide, if measurements exist for more than one type of assay, an auxiliary module, REPORT, estimates the intake by applying weights assigned in the nuclide file for each assay. Bioassay data and computed results (estimates of intake and committed dose equivalent) are stored in separate data bases, and the bioassay measurements used to compute a given result can be identified. The REPORT module creates a file containing committed effective dose equivalent for each individual that can be combined with the individual's external exposure.

  18. Book Review: Bioassays with Arthropods: 2nd Edition

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The technical book "Bioassays with Arthropods: 2nd Edition" (2007. Jacqueline L. Robertson, Robert M. Russell, Haiganoush K, Preisler and N. E. Nevin, Eds. CRC Press, Boca Raton, FL, 224 pp.) was reviewed for the scientific readership of the peer-reviewed publication Journal of Economic Entomology. ...

  19. US Army Radiological Bioassay and Dosimetry: The RBD software package

    SciTech Connect

    Eckerman, K.F.; Ward, R.C.; Maddox, L.B.

    1993-01-01

    The RBD (Radiological Bioassay and Dosimetry) software package was developed for the U. S. Army Material Command, Arlington, Virginia, to demonstrate compliance with the radiation protection guidance 10 CFR Part 20 (ref. 1). Designed to be run interactively on an IBM-compatible personal computer, RBD consists of a data base module to manage bioassay data and a computational module that incorporates algorithms for estimating radionuclide intake from either acute or chronic exposures based on measurement of the worker`s rate of excretion of the radionuclide or the retained activity in the body. In estimating the intake,RBD uses a separate file for each radionuclide containing parametric representations of the retention and excretion functions. These files also contain dose-per-unit-intake coefficients used to compute the committed dose equivalent. For a given nuclide, if measurements exist for more than one type of assay, an auxiliary module, REPORT, estimates the intake by applying weights assigned in the nuclide file for each assay. Bioassay data and computed results (estimates of intake and committed dose equivalent) are stored in separate data bases, and the bioassay measurements used to compute a given result can be identified. The REPORT module creates a file containing committed effective dose equivalent for each individual that can be combined with the individual`s external exposure.

  20. BENTHIC INVERTEBRATE BIOASSAYS WITH TOXIC SEDIMENT AND PORE WATER

    EPA Science Inventory

    The relative sensitivities of bioassays to determine the toxicity of sediments were investigated and three methods of making the sample dilutions required to generate dose-response relationships were compared. he assays studied were: (a) Microtox, a 15-min assay of Photobacterium...

  1. Shape-encoded silica microparticles for multiplexed bioassays.

    PubMed

    Kim, Lily Nari; Kim, Mira; Jung, Keumsim; Bae, Hyung Jong; Jang, Jisung; Jung, Yushin; Kim, Jiyun; Kwon, Sunghoon

    2015-08-01

    Shape-encoded silica microparticles for use in multiplexed bioassays were fabricated by using optofluidic maskless lithography (OFML) and tetraethylorthosilicate (TEOS) polymerization. These encoded silica microparticles exhibit excellent bioconjugation properties and negligible non-specific analyte adsorption. Encoded silica microparticles could be useful in a wide variety of applications, including DNA- and protein-based diagnostics. PMID:26125980

  2. Microplate Bioassay for Determining Substrate Selectivity of "Candida rugosa" Lipase

    ERIC Educational Resources Information Center

    Wang, Shi-zhen; Fang, Bai-shan

    2012-01-01

    Substrate selectivity of "Candida rugosa" lipase was tested using "p"-nitrophenyl esters of increasing chain length (C[subscript 1], C[subscript 7], C[subscript 15]) using the high-throughput screening method. A fast and easy 96-well microplate bioassay was developed to help students learn and practice biotechnological specificity screen. The…

  3. Sensitive bioassay for detection of biologically active ricin in food

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential use of ricin as an agent of biological warfare highlights the need to develop fast and effective methods to detect biologically active ricin. The current “gold standard” for ricin detection is an in vivo mouse bioassay; however, this method is not practical to test on a large number of...

  4. Correction of spray concentration and bioassay cage penetration data

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field trials were conducted to demonstrate the need for correcting sampled spray concentration data for sampler collection efficiencies and estimated spray exposure levels in mosquito bioassays for cage interference effects. A large spray block was targeted with aerial spray treatments of etofenpro...

  5. 1. VIEW IN ROOM 125, BIOASSAY LABORATORY, SHOWN IS THE ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    1. VIEW IN ROOM 125, BIOASSAY LABORATORY, SHOWN IS THE FIRST STEP IN A SIX-STEP PROCESS TO ANALYZE URINE SAMPLES FOR PLUTONIUM AND URANIUM CONTAMINATION. IN THIS STEP, NITRIC ACID IS ADDED TO SAMPLE, AND THE SAMPLE IS BOILED DOWN TO A WHITE POWDER. - Rocky Flats Plant, Health Physics Laboratory, On Central Avenue between Third & Fourth Streets, Golden, Jefferson County, CO

  6. Medium-term bioassays for carcinogenicity of chemical mixtures.

    PubMed Central

    Ito, N; Imaida, K; Hirose, M; Shirai, T

    1998-01-01

    Carcinogenic effects of chemical mixtures were examined with a medium-term liver bioassay for carcinogens or a multiorgan medium-term bioassay using male F344 rats. In the medium-term liver bioassay, rats were initially treated with diethylnitrosamine (DEN) at 200 mg/kg body weight, i.p.; after 2 weeks they received chemical mixtures such as 10 different heterocyclic amines at one-tenth or one-hundredth the dose levels used in carcinogenicity studies and the mixtures of 20 different pesticides, each at acceptable daily intake (ADI) levels or a mixture of 100 times ADI levels. All animals were subjected to two-thirds partial hepatectomy at week 3 and were sacrificed at week 8. The number and areas of glutathione S-transferase placental form (GST-P) positive foci (preneoplastic lesions in the liver) were compared between respective groups. When 10 heterocyclic amines were mixed in the diet at one-tenth dose level, clear synergism was observed, but no combined effects were evident with the one-hundredth dose levels. In the pesticide experiment, treatment of rats with the 20-pesticide mixture at the ADI dose level did not enhance GST-P-positive foci. In contrast, a mixture of 100 times the ADI significantly increased those values. In a multiorgan bioassay of 28 weeks, mixtures of 40 high-volume compounds and 20 pesticides (suspected carcinogens) added together at their respective ADI levels did not enhance carcinogenesis in any organs initiated by five different carcinogens (DEN, N-methylnitrosourea, dimethylhydrazine, N-butyl-N-(4-hydroxybutyl)nitrosamine, and dihydroxy-di-n-propylnitrosamine) in combination. The combination effect of low dietary levels of five antioxidants, butylated hydroxyanisole, caffeic acid, sesamol, 4-methoxyphenol, and catechol, were also examined using the multiorgan bioassay. The incidence of forestomach papillomas was significantly increased only in the combination group and the results indicate that combination of the five antioxidants can

  7. Preparation and characterization of bio-nanocomposite films of agar and silver nanoparticles: laser ablation method.

    PubMed

    Rhim, Jong-Whan; Wang, Long-Feng; Lee, Yonghoon; Hong, Seok-In

    2014-03-15

    Silver nanoparticles (AgNPs) were prepared by a laser ablation method and composite films with the AgNPs and agar were prepared by solvent casting method. UV-vis absorbance test and transmission electron microscopy (TEM) analysis results revealed that non-agglomerated spherical AgNPs were formed by the laser ablation method. The surface color of the resulting agar/AgNPs films exhibited the characteristic plasmonic effect of the AgNPs with the maximum absorption peaks of 400-407 nm. X-ray diffraction (XRD) test results also exhibited characteristic AgNPs crystals with diffraction peaks observed at 2θ values of 38.39°, 44.49°, and 64.45°, which were corresponding to (111), (200), and (220) crystallographic planes of face-centered cubic (fcc) silver crystals, respectively. Thermogravimetric analysis (TGA) results showed that thermal stability of the agar/AgNPs composite films was increased by the inclusion of metallic silver. Water vapor barrier properties and surface hydrophobicity of the agar/AgNPs films increased slightly with the increase in AgNPs content but they were not statistically significant (p>0.05), while mechanical strength and stiffness of the composite films decreased slightly (p<0.05). The agar/AgNPs films exhibited distinctive antimicrobial activity against both Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli O157:H7) bacterial pathogens. PMID:24528754

  8. 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

  9. Characterization of bionanocomposite films prepared with agar and paper-mulberry pulp nanocellulose.

    PubMed

    Reddy, Jeevan Prasad; Rhim, Jong-Whan

    2014-09-22

    Crystallized nanocellulose (CNC) was separated from paper-mulberry (Broussonetia kazinoki Siebold) bast pulp by sulfuric acid hydrolysis method and they were blended with agar to prepare bionanocomposite films. The effect of CNC content (1, 3, 5 and 10 wt% based on agar) on the mechanical, water vapor permeability (WVP), and thermal properties of the nanocomposites were studied. Changes of the cellulose fibers in structure, morphology, crystallinity, and thermal properties of the films were evaluated using FT-IR, TEM, SEM, XRD, and TGA analysis methods. The CNC was composed of fibrous and spherical or elliptic granules of nano-cellulose with sizes of 50-60 nm. Properties of agar film such as mechanical and water vapor barrier properties were improved significantly (p<0.05) by blending with the CNC. The tensile modulus and tensile strength of agar film increased by 40% and 25%, respectively, in the composite film with 5 wt% of CNC, and the WVP of agar film decreased by 25% after formation of nanocomposite with 3 wt% of CNC. The CNC obtained from the paper-mulberry bast pulp can be used as a reinforcing agent for the preparation of bio-nanocomposites, and they have a high potential for the development of completely biodegradable food packaging materials. PMID:24906782

  10. 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. PMID:26043937

  11. 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

  12. 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

  13. Dioxin analysis in water by using a passive sampler and CALUX bioassay.

    PubMed

    Addeck, Amr; Croes, Kim; Van Langenhove, Kersten; Denison, Michael; Elskens, Marc; Baeyens, Willy

    2012-01-15

    Passive sampling of organic pollutants is a new trend in environmental monitoring and analysis. Passive samplers are being developed to overcome the drawbacks of the conventional snapshot sampling approach. The ceramic toximeter is a promising passive sampler for monitoring dioxin-contaminated surface and ground waters. It consists of an alumina cylinder lined with a thin coating of titania and a pore diameter of 0.05 μm. The cylinder serves as a diffusion barrier limiting the analyte transport to molecular diffusion only, as well as a container for a selective trapping material of a high capacity and affinity towards the chemical(s) of concern. The cylinder is closed from both sides with PTFE caps. The ceramic toximeter was filled with activated carbon as the trapping material and has been tested in vitro for the sampling of dioxin-contaminated water. In addition, the utilization of the CALUX bioassay technique for analyzing the trapped dioxin has greatly reduced the time and costs for dioxin scanning in aqueous media. Exposure times varied between 1 and 7 days in a solution of 1.35 ng-TCDDL(-1) (TCDD is 2,3,7,8-tetrachlorodibenzodioxin). The mean effective molecular diffusion coefficient of TCDD in the toximeter amounts to 11.9×10(-6)m(2)d(-1) while the minimum concentration detectable in an aquatic system after 30 days of exposure amounts to 0.89 pg-TCDDL(-1). PMID:22265472

  14. 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

  15. Simple protocol for secondary school hands-on activity: Electrophoresis of pre-stained nucleic acids on agar-agar borate gels.

    PubMed

    Britos, Leticia; Goyenola, Guillermo; Oroño, Silvia Umpiérrez

    2004-09-01

    An extremely simple, inexpensive, and safe method is presented, which emulates nucleic acids isolation and electrophoretic analysis as performed in a research environment, in the context of a secondary school hands-on activity. The protocol is amenable to an interdisciplinary approach, taking into consideration the electrical and chemical parameters of the electrophoretic system. Furthermore, the laboratory is framed in a more comprehensive pedagogical setting, which addresses the methodological aspects of a pivotal scientific enterprise such as the Human Genome Project. In this setting, the hands-on activity is complemented with animations, paper models, and discussions. Additionally, our results indicate that the use of borate buffer and agar-agar gels suits many of the experiments included in college-level laboratory activities, which currently make use of more expensive agarose gels and TBE or TAE buffers. PMID:21706751

  16. Genotoxicity of leachates from a landfill using three bioassays.

    PubMed

    Cabrera, G L; Rodriguez, D M

    1999-05-19

    In the city of Queretaro, around 500 tons of solid wastes are produced everyday and are deposited in a landfill. This is the result of social and economic activities of human beings or from their normal physiological functions. As a result of rain, leachates are produced, which, if not handled and treated correctly, may pollute the underground water. Among the bioassays developed for the detection of mutagenicity in environmental pollutants, plant systems have been proven to be sensitive, cheap, and effective. The purpose of this study was to determine the presence of genotoxic agents in the leachates of the landfill of the city using three bioassays: Tradescantia-micronucleus (Trad-MCN), Tradescantia stamen hair mutations (Trad-SHM) and Allium root anaphase aberrations (AL-RAA) and make a comparison of the results in the three assays. Leachates were sampled during both the dry and rainy seasons. Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the leachates. Three replicates of each sample were analyzed in each of the three bioassays. As expected the samples of leachates collected during the dry season showed a higher genotoxicity than those collected during the rainy season. In conclusion, there are substances present in the leachates capable of inducing genotoxicity in the plant assays. On the other hand, the plant assays showed different degrees of sensitivity: the more sensitive was the Trad-MCN bioassay and the less sensitive the Trad-SHM assay. Therefore, when analyzing environmental pollutants it is recommended to use a battery of bioassays. PMID:10350599

  17. Comparison of a new, bismuth-iron-sulfite-cycloserine agar for isolation of Clostridium perfringens with the tryptose-sulfite-cycloserine and blood agars.

    PubMed

    Gubash, S M; Ingham, L

    1997-02-01

    A new differential and selective, bismuth-iron-sulfite-cycloserine (BISC) medium, for isolation and enumeration of Clostridium perfringens from food and feces, was developed. The medium was compared with the widely-used tryptose-sulfite-cycloserine (TSC) medium and blood agar (BA) in recovering actively growing cells, cold- (refrigerated and frozen) stressed, and heat-stressed C. perfringens cells, and heat-activated spores from human feces. Both selective media were satisfactory in recovering actively growing cells and heat-activated spores of C. perfringens. Both were inferior to non-inhibitory blood agar in recovering heat or cold-stressed cells. The advantages of the new BISC medium over the TSC medium were: elimination of the need to prepare pour- or overlay-agar plates, which simplified inoculation of specimens on the medium and simplified the subcultures of colonies for confirmatory identification. All colonies of C. perfringens developed on BISC were black or dark gray. This was contrary to TSC medium, which gave, on average, 39.6% of white colonies when inoculated with the pure cultures of C. perfringens. PMID:9084113

  18. 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

  19. Blood agar to detect virulence factors in tap water heterotrophic bacteria.

    PubMed Central

    Payment, P; Coffin, E; Paquette, G

    1994-01-01

    Cytolytic colonies were found in 57% of tap water samples, and up to 6% of samples were found to contain bacteria having three or more virulence factors. The factors evaluated were cytotoxicity, hemolysis, cell adherence, and cell invasiveness. Overall, 17% of the samples contained cytolytic colonies that were adherent and hemolytic. Among the media tested, tryptic soy agar with sheep blood (incubated at 35 degrees C for 48 h) was the best medium for the detection of cytolytic colonies. Of the colonies growing on this medium, 13% were cytolytic, whereas on medium R2A, less than 3% were cytolytic. Furthermore, when tryptic soy agar with blood was used, 24% of the samples contained colonies with at least three virulence factors whereas only 5% were positive with R2A. Routine monitoring by using tryptic soy agar with sheep blood is suggested as an appropriate procedure for the detection of bacteria with pathogenic potential in drinking water. PMID:8017913

  20. [DNA and chemical analyses of commercial fly agaric-related products].

    PubMed

    Maruyama, Takuro; Kawahara, Nobuo; Fukiharu, Toshimitsu; Yokoyama, Kazumasa; Makino, Yukiko; Goda, Yukihiro

    2005-04-01

    Since June 6, 2002, psilocin and psilocybin-containing fungi (commonly called "magic mushrooms") have been regulated by the Narcotics and Psychotropics Control Law in Japan. However, various fly agaric-related products are now entering the Japanese market via the internet. In this study, fly agaric-related products available in this way were investigated for raw materials by DNA analysis and for additives by chemical analysis. Nucleotide sequence analysis of the mitochondrial 12S rDNA region suggested that these fly agaric-related products originate from A. muscaria or A. muscaria var. persicina. Furthermore, they were classified into three strains based on the ITS2-LSU nucleotide sequence. Harmine derivatives and/or tryptamine derivatives were detected in some of these products by LC/MS analysis. In accordance with this, the matK gene of Peganum harmala was found in all of the harmine derivative-containing samples. PMID:16018591

  1. 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.

  2. Growth kinetics of Salmonella enterica in Hajna tetrathionate broth, Rappaport broth and modified semisolid Rappaport agar

    PubMed Central

    FUJIHARA, Masatoshi; TABUCHI, Hiroyuki; UEGAKI, Kaho

    2015-01-01

    To determine the appropriate method for isolating Salmonella enterica, we compared the growth of S. enterica serovars using three selective enrichment media. S. enterica was more successfully isolated from artificially contaminated fecal samples after enrichment in Hajna tetrathionate broth or modified semisolid Rappaport agar than in Rappaport broth. Since most bacteria (other than motile S. enterica) do not migrate on modified semisolid Rappaport agar, the growth characteristics of S. enterica can be interpreted easily and quickly. Two S. enterica isolates did not migrate on modified semisolid Rappaport agar, but did grow in Hajna tetrathionate broth, which suggests that the combined use of these selective enrichment media is appropriate for isolating S. enterica. PMID:26498402

  3. 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. PMID:24554750

  4. Assessment of Etest as an Alternative to Agar Dilution for Antimicrobial Susceptibility Testing of Neisseria gonorrhoeae

    PubMed Central

    Taylor, Thomas H.; Pettus, Kevin; Trees, David

    2014-01-01

    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. PMID:24554750

  5. A Simple Experiment for Visualizing Diffusion

    ERIC Educational Resources Information Center

    Helseth, L. E.

    2011-01-01

    We propose a simple and fascinating experiment for studying diffusion in gels using a pH-sensitive dye. By doping agar with methyl red, we obtain a gel which rapidly reacts to changes in pH by changing its absorption spectrum. The pH gradients can be followed using a digital camera, and we demonstrate here that the pH-sensitive colour changes can…

  6. Serogroup identification of Neisseria meningitidis: comparison of an antiserum agar method with bacterial slide agglutination.

    PubMed Central

    Craven, D E; Frasch, C E; Robbins, J B; Feldman, H A

    1978-01-01

    A serum agar method for serogrouping Neisseria meningitidis is described and compared with conventional bacterial slide agglutination. There was 93% agreement for 300 strains examined individually by each method. Among strains from serogroups A, B, C, Y, and W135, there was 100% correlation, whereas strains from serogroup 29E (Z') had only 67% correlation. The serum agar method was rapid, as well as easy to perform and interpret. The potential benefits of this method for epidemiological studies and reference laboratories processing large numbers of meningococcal isolates are emphasized. Images PMID:96123

  7. Casein Agar: a Useful Medium for Differentiating Candida dubliniensis from Candida albicans

    PubMed Central

    Mosca, Christian O.; Moragues, María D.; Llovo, José; Al Mosaid, Asmaa; Coleman, David C.; Pontón, José

    2003-01-01

    Production of chlamydospores on casein agar at 24°C for 48 h provides a simple means for differentiating Candida dubliniensis from Candida albicans based on chlamydospore production. Of 109 C. dubliniensis isolates tested on this medium, 106 (97.2%) produced abundant chlamydospores and three produced few chlamydospores. In contrast, of the 120 C. albicans isolates tested, 111 (92.5%) failed to produce any chlamydospores, whereas the remaining nine isolates produced few chlamydospores. These findings indicate that abundant chlamydospore production on casein agar is a useful test for discriminating between C. dubliniensis and C. albicans. PMID:12624062

  8. BioAssay Ontology (BAO): a semantic description of bioassays and high-throughput screening results

    PubMed Central

    2011-01-01

    Background High-throughput screening (HTS) is one of the main strategies to identify novel entry points for the development of small molecule chemical probes and drugs and is now commonly accessible to public sector research. Large amounts of data generated in HTS campaigns are submitted to public repositories such as PubChem, which is growing at an exponential rate. The diversity and quantity of available HTS assays and screening results pose enormous challenges to organizing, standardizing, integrating, and analyzing the datasets and thus to maximize the scientific and ultimately the public health impact of the huge investments made to implement public sector HTS capabilities. Novel approaches to organize, standardize and access HTS data are required to address these challenges. Results We developed the first ontology to describe HTS experiments and screening results using expressive description logic. The BioAssay Ontology (BAO) serves as a foundation for the standardization of HTS assays and data and as a semantic knowledge model. In this paper we show important examples of formalizing HTS domain knowledge and we point out the advantages of this approach. The ontology is available online at the NCBO bioportal http://bioportal.bioontology.org/ontologies/44531. Conclusions After a large manual curation effort, we loaded BAO-mapped data triples into a RDF database store and used a reasoner in several case studies to demonstrate the benefits of formalized domain knowledge representation in BAO. The examples illustrate semantic querying capabilities where BAO enables the retrieval of inferred search results that are relevant to a given query, but are not explicitly defined. BAO thus opens new functionality for annotating, querying, and analyzing HTS datasets and the potential for discovering new knowledge by means of inference. PMID:21702939

  9. Thin agar film for enhanced fungal growth and microscopic viewing in a new sealable fungal culture case.

    PubMed

    Hill, D R

    1996-09-01

    This project was undertaken to find ways to enhance fungus colony maturation, to make viewing of fungal cultures easier, and to reduce disruption of the fungal structures to be observed for identification. Accordingly, a technique using a thin (0.2-mm) agar film that avoids problems inherent in traditional methods of fungal culture and identification was developed. In addition, to accommodate the 0.2-mm layer of agar film and a contiguous thicker 4-mm section of agar, a sealable fungal culture case that fits within microscope stage calipers and under the objective lenses was invented. The growth and identification of 28 organisms were evaluated in the sealable fungal culture cases and on double-pour agar plates by using potato dextrose agar in both. Compared with results obtained with the double-pour agar plates (rated as "good"), fungal growth and identification with the sealable fungal culture case were superior (rated as "excellent") (P < 0.05, chi-square test). The thin agar film limits excessive mycelial growth, while it often promotes complete sporulation or other forms of maturation of the fungal colony. More importantly, the thin agar film allows direct microscopic viewing of the developing fungal colonies. The portion of the sealable fungal culture case with the 4-mm layer of agar can be used for evaluation of colony pigment and texture. In conclusion, this new sealable fungal culture case allows direct viewing and earlier fungal species identification with greater intrinsic safety. PMID:8862573

  10. Comparison of Etest, disk diffusion, and broth macrodilution for in vitro susceptibility testing of Rhodococcus equi.

    PubMed

    Berghaus, Londa J; Giguère, Steeve; Guldbech, Kristen; Warner, Eleanor; Ugorji, Ukachi; Berghaus, Roy D

    2015-01-01

    MICs of erythromycin, clarithromycin, azithromycin, rifampin, gentamicin, and doxycycline against 101 isolates of Rhodococcus equi were determined by broth macrodilution, disk diffusion, and Etest. Categorical agreement ranged between 85.1 and 100%. Overall, the agreement between Etest and disk diffusion was better than the agreement between broth macrodilution and the agar-based methods. PMID:25378571

  11. Comparison of Etest, Disk Diffusion, and Broth Macrodilution for In Vitro Susceptibility Testing of Rhodococcus equi

    PubMed Central

    Berghaus, Londa J.; Guldbech, Kristen; Warner, Eleanor; Ugorji, Ukachi; Berghaus, Roy D.

    2014-01-01

    MICs of erythromycin, clarithromycin, azithromycin, rifampin, gentamicin, and doxycycline against 101 isolates of Rhodococcus equi were determined by broth macrodilution, disk diffusion, and Etest. Categorical agreement ranged between 85.1 and 100%. Overall, the agreement between Etest and disk diffusion was better than the agreement between broth macrodilution and the agar-based methods. PMID:25378571

  12. Pullulan encapsulation of labile biomolecules to give stable bioassay tablets.

    PubMed

    Jahanshahi-Anbuhi, Sana; Pennings, Kevin; Leung, Vincent; Liu, Meng; Carrasquilla, Carmen; Kannan, Balamurali; Li, Yingfu; Pelton, Robert; Brennan, John D; Filipe, Carlos D M

    2014-06-10

    A simple and inexpensive method is reported for the long-term stabilization of enzymes and other unstable reagents in premeasured quantities in water-soluble tablets (cast, not compressed) made with pullulan, a nonionic polysaccharide that forms an oxygen impermeable solid upon drying. The pullulan tablets dissolve in aqueous solutions in seconds, thereby facilitating the easy execution of bioassays at remote sites with no need for special reagent handling and liquid pipetting. This approach is modular in nature, thus allowing the creation of individual tablets for enzymes and their substrates. Proof-of-principle demonstrations include a Taq polymerase tablet for DNA amplification through PCR and a pesticide assay kit consisting of separate tablets for acetylcholinesterase and its chromogenic substrate, indoxyl acetate, both of which are highly unstable. The encapsulated reagents remain stable at room temperature for months, thus enabling the room-temperature shipping and storage of bioassay components. PMID:24764260

  13. A New Bioassay for Auxins and Cytokinins 1

    PubMed Central

    Boerjan, Wout; Genetello, Chris; Van Montagu, Marc; Inzé, Dirk

    1992-01-01

    The authors have developed a sensitive bioassay that can be used to detect auxins as well as cytokinins. The bioassay is based on the expression in transformed tobacco (Nicotiana tabacum) mesophyll protoplasts of a chimeric gene, consisting of the upstream sequences of the Agrobacterium tumefaciens gene 5, coupled to the coding sequence of the β-glucuronidase. The expression of this gene is induced by the presence of both auxin and cytokinin in the culture medium. Using this assay, indole-3-acetic acid was detected at 5 × 10−8 molar, whereas trans-zeatin could be detected at 5 × 10−11 molar. The assay can be performed in microtiter plates, allowing numerous samples to be analyzed simultaneously. Only 2.5 × 105 protoplasts are required for one individual assay in 250 microliters of culture medium and for qualitative results, the reaction is readily visualized by ultraviolet light. ImagesFigure 3Figure 4Figure 6 PMID:16668975

  14. Bioassay studies to determine OTEC's effect on phytoplankton activity

    SciTech Connect

    Carmiggelt, C.J.W.; Hartwig, E.O.; Commins, M.L.; Horne, A.J.

    1982-09-01

    The effect of artificially upwelled water (800m) on phytoplankton from 25m and 100m was simulated using five day bioassays. The results show that some enhancement of the phytoplankton populations in the receiving waters due to upwelling is likely to occur. The very small phytoplankton (< 5 um) are most important in this response. The magnitude of the biostimulation cannot be predicted from this study. Ammonia leaks, spills, and venting are probable in an operating OTEC plant. The bioassays show that additions of ammonia will produce biostimulaton only when the P/N ratio indicates nitrogen limitation. In the Hawaiian waters sampled N-limitation was not always present and varied with depth. No nitrogen fixation was detected. The magnitude of stimulation due to ammonia alone was generally less than the addition of upwelled water which is a more complete nutrient mixture.

  15. Applying Agar's Concept of "Languaculture" to Explain Asian Students' Experiences in the Australian Tertiary Context

    ERIC Educational Resources Information Center

    Norris, Lindy; Tsedendamba, Nara

    2015-01-01

    This paper reports part of a broader qualitative case study of Asian students "translation" (Agar, 2006) to study in an Australian university. The paper is concerned with the experiences of eight participants and their involvement in a training programme in the use of language learning strategies (LLS) to support their engagement with…

  16. Development of hyperspectral imaging technique for salmonella enteritidis and typhimurium on agar plates

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Salmonella is a common cause of foodborne disease resulting from the consumption of contaminated food products. Although a direct plating method is widely used for presumptive positive screening of pathogenic Salmonella colonies on agar plates, it is labor-intensive, time-consuming and also prone to...

  17. THE MICROGARDENING COOKBOOK, DIRECTIONS FOR PREPARING DISHES AND TUBES OF STERILE NUTRIENT AGAR.

    ERIC Educational Resources Information Center

    CHANDLER, MARION N.

    THIS BOOKLET WAS PREPARED FOR TEACHER USE IN ASSOCIATION WITH THE ELEMENTARY SCIENCE STUDY UNIT "MICROGARDENING." IT CONTAINS DIRECTIONS FOR PREPARING CULTURE DISHES AND TUBES OF NUTRIENT STERILE AGAR FOR FUNGAL AND/OR BACTERIAL GROWTH. IT INCLUDES (1) LISTS OF NEEDED SUPPLIES AND EQUIPMENT, (2) DIRECTIONS FOR THE PREPARATION AND STERILIZATION OF…

  18. Mercury in fruiting bodies of Fly Agaric Amanita muscaria (L.: Fr.) Pers. collected from Poland

    NASA Astrophysics Data System (ADS)

    Falandysz, J.; Lipka, K.

    2003-05-01

    Total mercury concentrations were determined in the fruiting bodies of Fly Agaric Amanita muscaria (L.: FL) Pers. and underlying soil substrate collected from several sites in Poland in 1993-2000 to evaluate mercury status as contaminant and bioindicating features of this species. The samples were collected from the spatially distant sites such as: Zaborski Landscape Park, Mierzeja Wiślana Landscape Park, Wdzydzki Landscape Park, Borecka Forest, Tucholskie Forest, Wieluńska Upland, the communities of Gubin, Manowo, Lubiana and Morag. Total mercury content of caps and stalks of Fly agaric varied widely depending on the sites examined. The range of the mean mercury concentrations for all 17 sites was between 96±10 and 1900±1400 ng/g dry wt for the caps and between 6l±32 and 920±760 ng/g dry wt for the stalks, while between 4.4±3.1 and 150±20 ng/g were noted for soil substrate samples from 9 sites examined. Fly agaric independently of the site examined showed relatively good capacity to accumulate total mercury and BCF values varied between 16±10 and 74±15 for the caps and between 11±8 and 42±10 for the stalks. Nevertheless, relatively high bioconcentration potential of mercury by Fly agaric seems to be specific for that species and under soil mercury concentrations noted no bioindication properties of this mushroom could be observed.

  19. Seasonal variation in the biomass and agar yield from Gracilaria cervicornis and Hydropuntia cornea from Brazil.

    PubMed

    Marinho-Soriano, E; Silva, T S; Moreira, W S

    2001-04-01

    Seasonality of biomass and agar yield from two agarophytes (G. cervicornis and H. cornea) was determined. The biomass from G. cervicornis was higher (390 g m-2) during the dry season and lower during the rainy season (129 g m-2). The data analysis for G. cervicornis revealed a significant seasonal variation (P < 0.05). H. cornea did not show a clear seasonal variation and was present only from March to August. The peak in biomass for this species was recorded in April (383 g m-2) and was significantly different from the other months (P < 0.05). The agar yield for G. cervicornis varied from 11% to 20%, with generally higher values recorded during the dry season. The agar yield showed a highly significant variation (P < 0.001). Agar yield from H. cornea ranged from 29% to 41%, with a peak recorded in June. The results above indicate that H. cornea can be considered a good candidate for commercial use. PMID:11272017

  20. Radiation survival of murine and human melanoma cells utilizing two assay systems: monolayer and soft agar.

    PubMed Central

    Yohem, K. H.; Slymen, D. J.; Bregman, M. D.; Meyskens, F. L.

    1988-01-01

    The radiation response of murine and human melanoma cells assayed in bilayer soft agar and monolayer was examined. Cells from the murine melanoma Cloudman S91 CCL 53.1 cell line and three human melanoma cell strains (C8146C, C8161, and R83-4) developed in our laboratory were irradiated by single dose X-rays and plated either in agar or on plastic. D0 values were the same within 95% confidence intervals for cells from the human melanoma cell strains C8146C, C8161, and R83-4 but were dissimilar for the murine cell line CCL 53.1 Dq values were different for all cells studied. The shape of the survival curve for all four melanomas was not identical for cells assayed in soft agar versus cells grown on plastic. This would indicate that apparent radiosensitivity was influenced by the method of assay although there were no apparent consistent differences between the curves generated by monolayer or bilayer soft agar assays. PMID:3348949

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 sp...

  2. EFFECT OF IMPACT STRESS ON MICROBIAL RECOVERY ON AN AGAR SURFACE

    EPA Science Inventory

    Microbial stress due to the impaction of microorganisms onto an agar collection surface was studied experimentally. he relative recovery rates of aerosolized Pseudomonas fluorescens and Micrococcus luteus were determined as a function of the impaction velocity by using a moving a...

  3. A Method for Cell Culture and Maintenance of Ammonia-Oxidizing Archaea in Agar Stab.

    PubMed

    Chu, Yeon-Jin; Lee, Jin-Young; Shin, So-Ra; Kim, Geun-Joong

    2015-12-01

    Ammonia oxidizing archaea (AOA) are predominantly found and closely linked with geochemical cycling of nitrogen in non-extreme habitats. However, these strains have mainly been investigated using liquid cultures of enriched cells. Here, we provide an agar stab as a simple and reliable means of cultivating and maintaining AOA. PMID:26543273

  4. Evolutionary consequences of putative intra- and interspecific hybridization in agaric fungi

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Agaric fungi of the southern Appalachians including the Great Smoky Mountains National Park are often heterozygous for the rDNA internal transcribed spacer region (ITS) with >42% of collections showing some level of heterozygosity for indels and/or base-pair substitutions. For these collections, int...

  5. Fishmeal extract bile salt lactose agar--a differential medium for enteric bacteria.

    PubMed

    Subbannayya, K; Udayalaxmi, J; Anugraha, M

    2006-08-01

    Fishmeal extract bile salt lactose agar (FEBLA), a new differential medium for enteric bacteria was developed and evaluated for its ability to grow and differentiate lactose fermenters (LF) from non-lactose fermenters (NLF) in comparison with MacConkeys agar. Performance of FEBLA was at par with the latter. On FEBLA medium, the contrast between LF and NLF colonies was pronounced and Klebsiella pneumoniae produced more mucoid colonies than on MacConkeys agar (Hi Media). Unlike MacConkeys agar, a 24 h culture of K. pneumoniae cells on FEBLA were longer and thicker with abundant capsular material around the bacilli. Escherichia coli produced long and thick cells but only after 48h. No change in cell morphology was evident with regard to Salmonella typhi, S. paratyphi A, Shigella flexneri, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Citrobacter koseri and Acinetobacter baumannii. Performance of the medium was controlled using E. coli and S. flexneri. FEBLA is simple, cost effective and may be a suitable alternative in the preliminary identification of enteric bacteria. PMID:16924840

  6. 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.

  7. MODIFIED AGAR MEDIUM FOR DETECTING ENVIRONMENTAL SALMONELLAE BY THE MOST-PROBABLE-NUMBER METHOD

    EPA Science Inventory

    Salmonellae in the environment remain a potential source of disease. Low numbers of salmonellae have been detected and enumerated from environmental samples by most probable number methods that require careful colony selection from plated agar medium. A modified xylose lysine bri...

  8. Alternative plasticizers for the production of thermo-compressed agar films

    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...

  9. Investigation of dental alginate and agar impression materials as a brain simulant for ballistic testing.

    PubMed

    Falland-Cheung, Lisa; Piccione, Neil; Zhao, Tianqi; Lazarjan, Milad Soltanipour; Hanlin, Suzanne; Jermy, Mark; Waddell, J Neil

    2016-06-01

    Routine forensic research into in vitro skin/skull/brain ballistic blood backspatter behavior has traditionally used gelatin at a 1:10 Water:Powder (W:P) ratio by volume as a brain simulant. A limitation of gelatin is its high elasticity compared to brain tissue. Therefore this study investigated the use of dental alginate and agar impression materials as a brain simulant for ballistic testing. Fresh deer brain, alginate (W:P ratio 91.5:8.5) and agar (W:P ratio 81:19) specimens (n=10) (11×22×33mm) were placed in transparent Perspex boxes of the same internal dimensions prior to shooting with a 0.22inch caliber high velocity air gun. Quantitative analysis to establish kinetic energy loss, vertical displacement elastic behavior and qualitative analysis to establish elasticity behavior was done via high-speed camera footage (SA5, Photron, Japan) using Photron Fastcam Viewer software (Version 3.5.1, Photron, Japan) and visual observation. Damage mechanisms and behavior were qualitatively established by observation of the materials during and after shooting. The qualitative analysis found that of the two simulant materials tested, agar behaved more like brain in terms of damage and showed similar mechanical response to brain during the passage of the projectile, in terms of energy absorption and vertical velocity displacement. In conclusion agar showed a mechanical and subsequent damage response that was similar to brain compared to alginate. PMID:27131216

  10. 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

  11. Paper bioassay based on ceria nanoparticles as colorimetric probes.

    PubMed

    Ornatska, Maryna; Sharpe, Erica; Andreescu, Daniel; Andreescu, Silvana

    2011-06-01

    We report the first use of redox nanoparticles of cerium oxide as colorimetric probes in bioanalysis. The method is based on changes in the physicochemical properties of ceria nanoparticles, used here as chromogenic indicators, in response to the analyte. We show that these particles can be fully integrated in a paper-based bioassay. To construct the sensor, ceria nanoparticles and glucose oxidase were coimmobilized onto filter paper using a silanization procedure. In the presence of glucose, the enzymatically generated hydrogen peroxide induces a visual color change of the ceria nanoparticles immobilized onto the bioactive sensing paper, from white-yellowish to dark orange, in a concentration-dependent manner. A detection limit of 0.5 mM glucose with a linear range up to 100 mM and a reproducibility of 4.3% for n = 11 ceria paper strips were obtained. The assay is fully reversible and can be reused for at least 10 consecutive measurement cycles, without significant loss of activity. Another unique feature is that it does not require external reagents, as all the sensing components are fixed onto the paper platform. The bioassay can be stored for at least 79 days at room temperature while maintaining the same analytical performance. An example of analytical application was demonstrated for the detection of glucose in human serum. The results demonstrate the potential of this type of nanoparticles as novel components in the development of robust colorimetric bioassays. PMID:21524141

  12. A Bioassay System Using Bioelectric Signals from Small Fish

    NASA Astrophysics Data System (ADS)

    Terawaki, Mitsuru; Soh, Zu; Hirano, Akira; Tsuji, Toshio

    Although the quality of tap water is generally examined using chemical assay, this method cannot be used for examination in real time. Against such a background, the technique of fish bioassay has attracted attention as an approach that enables constant monitoring of aquatic contamination. The respiratory rhythms of fish are considered an efficient indicator for the ongoing assessment of water quality, since they are sensitive to chemicals and can be indirectly measured from bioelectric signals generated by breathing. In order to judge aquatic contamination accurately, it is necessary to measure bioelectric signals from fish swimming freely as well as to stably discriminate measured signals, which vary between individuals. However, no bioassay system meeting the above requirements has yet been established. This paper proposes a bioassay system using bioelectric signals generated from small fish in free-swimming conditions. The system records signals using multiple electrodes to cover the extensive measurement range required in a free-swimming environment, and automatically discriminates changes in water quality from signal frequency components. This discrimination is achieved through an ensemble classification method using probability neural networks to solve the problem of differences between individual fish. The paper also reports on the results of related validation experiments, which showed that the proposed system was able to stably discriminate between water conditions before and after bleach exposure.

  13. Improved bioassay for detecting autoinducer of Rhodovulum sulfidophilum

    NASA Astrophysics Data System (ADS)

    Terada, T.; Kikuchi, Y.; Umekage, S.

    2015-02-01

    Quorum sensing is a bacterial gene regulation system that enables prompt environmental adaptation in response to cell density. Quorum sensing is driven by an extracellularly secreted chemical signal called autoinducer. Gram-negative bacteria produce one or several types of N-acylhomoserine lactone (AHL) as autoinducers. Our previous study suggests that the gram-negative marine photosynthetic bacterium Rhodovulum sulfidophilum produces AHL in the early stationary phase and plays a role in maintaining the bacterial cell aggregates called "floc". We performed conventional bioassay to identify AHL production by using Chromobacterium violaceum VIR07, which produces violet pigment (violacein) in response to AHL with side chains ranging from C10 to C18 in length. However, we were not able to observe the violacein with good reproducibility, suggesting that inhibitory chemical compounds co-existed in the AHL extract. Therefore, we improved the extraction method; the ethyl acetate-extracted AHLs were fractionated by using reverse phase TLC. By using the re-extracted AHLs for the bioassay, we observed an obvious production of violacein. This result clearly indicates that R. sulfidophilum produces AHLs with side chains ranging from C10 to C18 in length and suggests the utility of improved bioassay for AHL detection.

  14. Novel bioassay using Bacillus megaterium to detect tetracycline in milk.

    PubMed

    Tumini, Melisa; Nagel, Orlando G; Molina, Pilar; Althaus, Rafael L

    2016-01-01

    Tetracyclines are used for the prevention and control of dairy cattle diseases. Residues of these drugs can be excreted into milk. Thus, the aim of this study was to develop a microbiological method using Bacillus megaterium to detect tetracyclines (chlortetracycline, oxytetracycline and tetracycline) in milk. In order to approximate the limits of detection of the bioassay to the Maximum Residue Limit (100μg/l) for milk tetracycline, different concentrations of chloramphenicol (0, 1000, 1500 and 2000μg/l) were tested. The detection limits calculated were similar to the Maximum Residue Limits when a bioassay using B. megaterium ATCC 9885 spores (2.8×10(8)spores/ml) and chloramphenicol (2000μg/l) was utilized. This bioassay detects 105μg/l of chlortetracycline, 100μg/l of oxytetracycline and 134μg/l of tetracycline in 5h. Therefore, this method is suitable to be incorporated into a microbiological multi-residue system for the identification of tetracyclines in milk. PMID:27131738

  15. Effect of impact stress on microbial recovery on an agar surface.

    PubMed

    Stewart, S L; Grinshpun, S A; Willeke, K; Terzieva, S; Ulevicius, V; Donnelly, J

    1995-04-01

    Microbial stress due to the impaction of microorganisms onto an agar collection surface was studied experimentally. The relative recovery rates of aerosolized Pseudomonas fluorescens and Micrococcus luteus were determined as a function of the impaction velocity by using a moving agar slide impactor operating over a flow rate range from 3.8 to 40 liters/min yielding impaction velocities from 24 to 250 m/s. As a reference, the sixth stage of the Andersen Six-Stage Viable Particle Sizing Sampler was used at its operating flow rate of 28.3 liters/min (24 m/s). At a collection efficiency of close to 100% for the agar slide impactor, an increase in sampling flow rate and, therefore, in impaction velocity produced a significant decline in the percentage of microorganisms recovered. Conversely, when the collection efficiency was less than 100%, greater recovery and lower injury rates occurred. The highest relative rate of recovery (approximately 51% for P. fluorescens and approximately 62% for M. luteus) was obtained on the complete (Trypticase soy agar) medium at 40 and 24 m/s (6.4 and 3.8 liters/min), respectively. M. luteus demonstrated less damage than P. fluorescens, suggesting the hardy nature of the gram-positive strain versus that of the gram-negative microorganism. Comparison of results from the agar slide and Andersen impactors at the same sampling velocity showed that recovery and injury due to collection depends not only on the magnitude of the impaction velocity but also on the degree to which the microorganisms may be embedded in the collection medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7747946

  16. Effect of impact stress on microbial recovery on an agar surface.

    PubMed Central

    Stewart, S L; Grinshpun, S A; Willeke, K; Terzieva, S; Ulevicius, V; Donnelly, J

    1995-01-01

    Microbial stress due to the impaction of microorganisms onto an agar collection surface was studied experimentally. The relative recovery rates of aerosolized Pseudomonas fluorescens and Micrococcus luteus were determined as a function of the impaction velocity by using a moving agar slide impactor operating over a flow rate range from 3.8 to 40 liters/min yielding impaction velocities from 24 to 250 m/s. As a reference, the sixth stage of the Andersen Six-Stage Viable Particle Sizing Sampler was used at its operating flow rate of 28.3 liters/min (24 m/s). At a collection efficiency of close to 100% for the agar slide impactor, an increase in sampling flow rate and, therefore, in impaction velocity produced a significant decline in the percentage of microorganisms recovered. Conversely, when the collection efficiency was less than 100%, greater recovery and lower injury rates occurred. The highest relative rate of recovery (approximately 51% for P. fluorescens and approximately 62% for M. luteus) was obtained on the complete (Trypticase soy agar) medium at 40 and 24 m/s (6.4 and 3.8 liters/min), respectively. M. luteus demonstrated less damage than P. fluorescens, suggesting the hardy nature of the gram-positive strain versus that of the gram-negative microorganism. Comparison of results from the agar slide and Andersen impactors at the same sampling velocity showed that recovery and injury due to collection depends not only on the magnitude of the impaction velocity but also on the degree to which the microorganisms may be embedded in the collection medium.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7747946

  17. 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

  18. Toxicity of copper-spiked sediments to Tubifex tubifex (Oligochaeta, Tubificidae): Comparison of the 28-day reproductive bioassay with an early-life-stage bioassay

    SciTech Connect

    Vecchi, M.; Pasteris, A.; Bonomi, G. . Dipt. di Biologia Evoluzionistica Sperimentale); Reynoldson, T.B. . National Water Research Inst.)

    1999-06-01

    Two sediment bioassay methods using Tubifex tubifex (Mueller, 1774) as the test species were compared. The first was an adult reproduction test, the second an early-life-stage survival test. The duration of both bioassays is 28 d and the amount of work required was similar; they may be useful alternatives to each other in different circumstances (e.g., the early life stage bioassay could be carried out with smaller volumes of sediment). The two bioassays were performed simultaneously on copper-spiked sediments. Sediments from two freshwater and two terrestrial sites were used; five separate, nonsimultaneous experiments were performed, one for each sediment or soil and a further experiment with soil with a good supplement. In the adult bioassay, there were large differences in the production of cocoons, eggs, and young among the control treatments of the five experiments. There were also major differences in the NOEC and LOEC for copper between the tested substrates. The early life stage bioassay appears to be less sensitive to copper toxicity than the adult reproductive bioassay since NOECs and LOECs are higher for early survival than for the most sensitive endpoints of the adult bioassay in three experiments out of five.

  19. In Vitro Biologic Activities of the Antimicrobials Triclocarban, Its Analogs, and Triclosan in Bioassay Screens: Receptor-Based Bioassay Screens

    PubMed Central

    Ahn, Ki Chang; Zhao, Bin; Chen, Jiangang; Cherednichenko, Gennady; Sanmarti, Enio; Denison, Michael S.; Lasley, Bill; Pessah, Isaac N.; Kültz, Dietmar; Chang, Daniel P.Y.; Gee, Shirley J.; Hammock, Bruce D.

    2008-01-01

    Background Concerns have been raised about the biological and toxicologic effects of the antimicrobials triclocarban (TCC) and triclosan (TCS) in personal care products. Few studies have evaluated their biological activities in mammalian cells to assess their potential for adverse effects. Objectives In this study, we assessed the activity of TCC, its analogs, and TCS in in vitro nuclear-receptor–responsive and calcium signaling bioassays. Materials and methods We determined the biological activities of the compounds in in vitro, cell-based, and nuclear-receptor–responsive bioassays for receptors for aryl hydrocarbon (AhR), estrogen (ER), androgen (AR), and ryanodine (RyR1). Results Some carbanilide compounds, including TCC (1–10 μM), enhanced estradiol (E2)-dependent or testosterone-dependent activation of ER- and AR-responsive gene expression up to 2.5-fold but exhibited little or no agonistic activity alone. Some carbanilides and TCS exhibited weak agonistic and/or antagonistic activity in the AhR-responsive bioassay. TCS exhibited antagonistic activity in both ER- and AR-responsive bioassays. TCS (0.1–10 μM) significantly enhanced the binding of [3H]ryanodine to RyR1 and caused elevation of resting cytosolic [Ca2+] in primary skeletal myotubes, but carbanilides had no effect. Conclusions Carbanilides, including TCC, enhanced hormone-dependent induction of ER- and AR-dependent gene expression but had little agonist activity, suggesting a new mechanism of action of endocrine-disrupting compounds. TCS, structurally similar to noncoplanar ortho-substituted poly-chlorinated biphenyls, exhibited weak AhR activity but interacted with RyR1 and stimulated Ca2+ mobilization. These observations have potential implications for human and animal health. Further investigations are needed into the biological and toxicologic effects of TCC, its analogs, and TCS. PMID:18795164

  20. A bioassay-guided fractionation scheme for characterization of new antibacterial compounds from Prosopis cineraria aerial parts

    PubMed Central

    Neghabi-Hajiagha, Mahdieh; Aliahmadi, Atousa; Taheri, Mohammad Reza; Ghassempour, Alireza; Irajian, Gholamreza; Rezadoost, Hassan; Feizabadi, Mohammad Mehdi

    2016-01-01

    Background and Objectives: Due to the importance of finding of new antibacterial agents, the antibacterial properties of Prosopis cineraria aerial parts were investigated using a bioassay guided fractionation scheme. Materials and Methods: The organic extract was prepared via maceration in methanol, followed by the fractionation using n-hexane and ethyl acetate. The MICs of fractions were determined against some human pathogenic bacteria using broth micro-dilution assay. The primary characterization and identification of bioactive substance(s) was based on a bio-autographical method using HPTLC and flash chromatography in parallel with agar overlay assays. Finally the exact mass of effective compound(s) was determined by LC-MS. Results: The best antibacterial activities were related to the ethyl acetate fraction. The effective antibacterial compound of the plant were 2 substances with molecular weight of 348 and 184 Dalton that inhibited the growth of assessed Gram positive bacteria with MIC values lower than 125 to 62.5 μg/ml synergistically. Conclusion: Further analysis using nuclear magnetic resonance could reveal the exact structure of these two antibacterial substances. These 2 effective antibacterial compounds could be applied as lead compound for synthesis of new antibacterial agents. PMID:27092218

  1. Microwave-accelerated bioassay technique for rapid and quantitative detection of biological and environmental samples.

    PubMed

    Mohammed, Muzaffer; Syed, Maleeha F; Aslan, Kadir

    2016-01-15

    Quantitative detection of molecules of interest from biological and environmental samples in a rapid manner, particularly with a relevant concentration range, is imperative to the timely assessment of human diseases and environmental issues. In this work, we employed the microwave-accelerated bioassay (MAB) technique, which is based on the combined use of circular bioassay platforms and microwave heating, for rapid and quantitative detection of Glial Fibrillary Acidic Protein (GFAP) and Shiga like toxin (STX 1). The proof-of-principle use of the MAB technique with the circular bioassay platforms for the rapid detection of GFAP in buffer based on colorimetric and fluorescence readouts was demonstrated with a 900W kitchen microwave. We also employed the MAB technique with a new microwave system (called the iCrystal system) for the detection of GFAP from mice with brain injuries and STX 1 from a city water stream. Control bioassays included the commercially available gold standard bioassay kits run at room temperature. Our results show that the lower limit of detection (LLOD) of the colorimetric and fluorescence based bioassays for GFAP was decreased by ~1000 times using the MAB technique and our circular bioassay platforms as compared to the commercially available bioassay kits. The overall bioassay time for GFAP and STX 1 was reduced from 4h using commercially available bioassay kits to 10min using the MAB technique. PMID:26356762

  2. Effect of heat treatment on the performance of tryptose-sulfite-cycloserine agar for enumeration of Clostridium perfringens.

    PubMed Central

    Brodsky, M H; Ciebin, B W

    1979-01-01

    Dissolving dehydrated tryptose-sulfite-cycloserine agar by only boiling or microwaving was found to inhibit Clostridium perfringens colony development in pour plates when compared with C. perfringens recovery in tryptose-sulfite-cycloserine agar prepared by autoclaving. Images PMID:225988

  3. Effect of lignin on water vapor barrier, mechanical, and structural properties of agar/lignin composite films.

    PubMed

    Shankar, Shiv; Reddy, Jeevan Prasad; Rhim, Jong-Whan

    2015-11-01

    Biodegradable composite films were prepared using two renewable resources based biopolymers, agar and lignin alkali. The lignin was used as a reinforcing material and agar as a biopolymer matrix. The effect of lignin concentration (1, 3, 5, and 10wt%) on the performance of the composite films was studied. In addition, the mechanical, water vapor barrier, UV light barrier properties, FE-SEM, and TGA of the films were analyzed. The agar/lignin films exhibited higher mechanical and UV barrier properties along with lower water vapor permeability compared to the neat agar film. The FTIR and SEM results showed the compatibility of lignin with agar polymer. The swelling ratio and moisture content of agar/lignin composite films were decreased with increase in lignin content. The thermostability and char content of agar/lignin composite films increased with increased lignin content. The results suggested that agar/lignin films have a potential to be used as a UV barrier food packaging material for maintaining food safety and extending the shelf-life of the packaged food. PMID:26271435

  4. Differentiating non-0157:H7 STEC serogroups from ground beef plated on agar media by hyperspetral imaging

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Introduction: The development of an assay to detect and confirm a positive non-O157:H7 isolate is challenging when mixed morphologically results are obtained from the serogroups growing on Rainbow agar. Rainbow agar is only claimed by the manufacturer to be very specific for E.coli O157:H7 strain...

  5. 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

  6. 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

  7. An abbreviated scheme for identification of Yersinia enterocolitica isolated from food enrichments on CIN (cefsulodin-irgasan-novobiocin) agar.

    PubMed

    Devenish, J A; Schiemann, D A

    1981-09-01

    An abbreviated procedure for the biochemical identification of Yersinia enterocolitica isolated from food enrichments on CIN (cefsulodin-irgasan-novobiocin) agar was investigated. A total of 170 colonies resembling Y. enterocolitica in colonial morphology and appearance on CIN agar were selected for identification using API strips. Ninety-three of these isolates were examined with the PathoTec ornithine decarboxylase, Voges-Proskauer, and urease test strips. The PathoTec urease strip alone was adequate for identification of all isolates of Y. enterocolitica. Christensen's urea agar was applied to the remaining 77 isolates and found less specific in the 1 isolate of Enterobacter agglomerans was urease positive along with 10 isolates of Y. enterocolitica. CIN agar is a highly specific medium for isolation of Y. enterocolitica, requiring only Kligler iron agar and urea slants for confirmation of presumptive colonies. PMID:7306881

  8. Modeling Antimicrobial Activity of Clorox(R) Using an Agar-Diffusion Test: A New Twist On an Old Experiment.

    ERIC Educational Resources Information Center

    Mitchell, James K.; Carter, William E.

    2000-01-01

    Describes using a computer statistical software package called Minitab to model the sensitivity of several microbes to the disinfectant NaOCl (Clorox') using the Kirby-Bauer technique. Each group of students collects data from one microbe, conducts regression analyses, then chooses the best-fit model based on the highest r-values obtained.…

  9. Enumeration of fecal Clostridium perfringens spores in egg yolk-free tryptose-sulfite-cycloserine agar.

    PubMed

    Hauschild, A H; Hilsheimer, R; Griffith, D W

    1974-03-01

    The Shahidi-Ferguson perfringens, tryptose-sulfite-cycloserine (TSC), and egg yolk-free TSC agars have been tested for their suitability to enumerate fecal spores of Clostridium perfringens. When these spores comprised at least 20% of the total anaerobe spores, equally accurate counts were obtained in the three media. With lower ratios of C. perfringens spores, the most accurate counts were obtained in egg yolk-free TSC agar. The median C. perfringens spore count of 60 normal fecal specimens was log 3.4/g. A nonmotile, sulfite- and nitrate-reducing Clostridium, not identifiable with any known clostridial species, was isolated from 14 out of 60 fecal specimans. It was not differentiated from C. perfringens in the nitrite motility test, but could be distinguished by its inability to liquefy gelatin. PMID:4363369

  10. Enumeration of Fecal Clostridium perfringens Spores in Egg Yolk-Free Tryptose-Sulfite-Cycloserine Agar

    PubMed Central

    Hauschild, A. H. W.; Hilsheimer, R.; Griffith, D. W.

    1974-01-01

    The Shahidi-Ferguson perfringens, tryptose-sulfite-cycloserine (TSC), and egg yolk-free TSC agars have been tested for their suitability to enumerate fecal spores of Clostridium perfringens. When these spores comprised at least 20% of the total anaerobe spores, equally accurate counts were obtained in the three media. With lower ratios of C. perfringens spores, the most accurate counts were obtained in egg yolk-free TSC agar. The median C. perfringens spore count of 60 normal fecal specimens was log 3.4/g. A nonmotile, sulfite- and nitrate-reducing Clostridium, not identifiable with any known clostridial species, was isolated from 14 out of 60 fecal specimans. It was not differentiated from C. perfringens in the nitrite motility test, but could be distinguished by its inability to liquefy gelatin. PMID:4363369

  11. Characterization of gelatin-agar based phase separated hydrogel, emulgel and bigel: a comparative study.

    PubMed

    Wakhet, Senggam; Singh, Vinay K; Sahoo, Saikat; Sagiri, Sai Sateesh; Kulanthaivel, Senthilguru; Bhattacharya, Mrinal K; Kumar, Naresh; Banerjee, Indranil; Pal, Kunal

    2015-02-01

    The current study describes the in-depth characterization of agar-gelatin based co-hydrogels, emulgels and bigels to have an insight about the differences in the properties of the formulations. Hydrogels have been extensively studied as vehicle for controlled drug release, whereas, the concept of emulgels and bigels is relatively new. The formulations were characterized by scanning electron microscopy, FTIR spectroscopy, XRD and mechanical properties. The biocompatibility and the ability of the formulations to be used as drug delivery vehicle were also studied. The scanning electron micrographs suggested the presence of internal phases within the agar-gelatin composite matrices of co-hydrogel, emulgel and bigel. FTIR and XRD studies suggested higher crystallinity of emulgels and bigels. Electrical impedance and mechanical stability of the emulgel and the bigel was higher than the hydrogel. The prepared formulations were found to be biocompatible and suitable for drug delivery applications. PMID:25672596

  12. Effect of Soybean Casein Digest Agar Lot on Number of Bacillus stearothermophilus Spores Recovered †

    PubMed Central

    Pflug, I. J.; Smith, Geraldine M.; Christensen, Ronald

    1981-01-01

    In recent years it has become increasingly apparent that Bacillus stearothermophilus spores are affected by various environmental factors that influence the performance of the spores as biological indicators. One environmental factor is the recovery medium. The effect of different lots of commercial soybean casein digest agar on the number of colony-forming units per plate was examined in two series of experiments: (i) several lots of medium from two manufacturers were compared in single experiments, and (ii) paired media experiments with four lots of medium were carried out and yielded three-point survivor curves. The results demonstrate that commercial soybean casein digest agar is variable on a lot-to-lot basis. The variation was lowest when recovering unheated or minimally heated spores and increased greatly with the severity of heating. PMID:16345822

  13. Effect of Soybean Casein Digest Agar Lot on Number of Bacillus stearothermophilus Spores Recovered.

    PubMed

    Pflug, I J; Smith, G M; Christensen, R

    1981-08-01

    In recent years it has become increasingly apparent that Bacillus stearothermophilus spores are affected by various environmental factors that influence the performance of the spores as biological indicators. One environmental factor is the recovery medium. The effect of different lots of commercial soybean casein digest agar on the number of colony-forming units per plate was examined in two series of experiments: (i) several lots of medium from two manufacturers were compared in single experiments, and (ii) paired media experiments with four lots of medium were carried out and yielded three-point survivor curves. The results demonstrate that commercial soybean casein digest agar is variable on a lot-to-lot basis. The variation was lowest when recovering unheated or minimally heated spores and increased greatly with the severity of heating. PMID:16345822

  14. Holographic diffusers

    NASA Astrophysics Data System (ADS)

    Wadle, Stephen; Wuest, Daniel; Cantalupo, John; Lakes, Roderic S.

    1994-01-01

    Holographic diffusers are prepared using silver halide (Agfa 8E75 and Kodak 649F) and photopolymer (Polaroid DMP 128 and DuPont 600, 705, and 150 series) media. It is possible to control the diffusion angle in three ways: by selection of the properties of the source diffuser, by control of its subtended angle, and by selection of the holographic medium. Several conventional diffusers based on refraction or scattering of light are examined for comparison.

  15. 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.

  16. Co-precipitation with PVP and Agar to Improve Physicomechanical Properties of Ibuprofen

    PubMed Central

    Maghsoodi, Maryam; Kiafar, Farhad

    2013-01-01

    Objective(s) : Ibuprofen is a problematic drug in tableting due to its viscoelastic properties. Additionally its high cohesivity results in low flowability. In this study, co-precipitation of ibuprofen with varying concentration of agar and PVP to optimize properties of Ibuprofen was carried out. Materials and Methods: Co-precipitates of ibuprofen- PVP or agar were prepared by solvent evaporation technique under vacuum condition. Differential scanning calorimetry (DSC), X -ray diffraction of powder (XRDP) and FT-IR spectroscopy were used to investigate the solid state characteristics of the co-precipitates. The dissolution behavior, flowability, particle size and compaction properties of various batches were also studied. Results: Co-precipitation of drug with agar led to a change in habit from needle to plate shape crystals, while drug –PVP co-precipitates had agglomerated structure and consisted of numerous crystals which had been aggregated together. The co-precipitates showed improved flow properties compared with ibuprofen alone. Precipitation of ibuprofen with these additives led to modification in the dissolution of the drug. Agar in 1% w/w improved slightly the dissolution rate of drug while PVP had a negative impact and led to reduction in the dissolution rate of drug to less than that of pure drug. The all obtained co-precipitates exhibited significantly improved tableting behavior compared with drug crystals alone. This may be due to this fact that, the polymer covering the drug particles increases and changes the nature of the surface area available for interparticulate bonds between particles. DSC, XRDP and FT-IR experiments showed that drug particles, in co-precipitates samples, did not undergo polymorphic modifications. Conclusion: The study highlights the influence of polymeric additives on crystallization process leading to modified performance. PMID:24250942

  17. Co-precipitation with PVP and Agar to Improve Physicomechanical Properties of Ibuprofen

    PubMed Central

    Maghsoodi, Maryam; Kiafar, Farhad

    2013-01-01

    Objective(s) : Ibuprofen is a problematic drug in tableting due to its viscoelastic properties. Additionally its high cohesivity results in low flowability. In this study, co-precipitation of ibuprofen with varying concentration of agar and PVP to optimize properties of Ibuprofen was carried out. Materials and Methods: Co-precipitates of ibuprofen- PVP or agar were prepared by solvent evaporation technique under vacuum condition. Differential scanning calorimetry (DSC), X -ray diffraction of powder (XRDP) and FT-IR spectroscopy were used to investigate the solid state characteristics of the co-precipitates. The dissolution behavior, flowability, particle size and compaction properties of various batches were also studied. Results: Co-precipitation of drug with agar led to a change in habit from needle to plate shape crystals, while drug –PVP co-precipitates had agglomerated structure and consisted of numerous crystals which had been aggregated together. The co-precipitates showed improved flow properties compared with ibuprofen alone. Precipitation of ibuprofen with these additives led to modification in the dissolution of the drug. Agar in 1% w/w improved slightly the dissolution rate of drug while PVP had a negative impact and led to reduction in the dissolution rate of drug to less than that of pure drug. The all obtained co-precipitates exhibited significantly improved tableting behavior compared with drug crystals alone. This may be due to this fact that, the polymer covering the drug particles increases and changes the nature of the surface area available for interparticulate bonds between particles. DSC, XRDP and FT-IR experiments showed that drug particles, in co-precipitates samples, did not undergo polymorphic modifications. Conclusion: The study highlights the influence of polymeric additives on crystallization process leading to modified performance. PMID:24250936

  18. Diffusion MRI

    NASA Astrophysics Data System (ADS)

    Fukuyama, Hidenao

    Recent advances of magnetic resonance imaging have been described, especially stressed on the diffusion sequences. We have recently applied the diffusion sequence to functional brain imaging, and found the appropriate results. In addition to the neurosciences fields, diffusion weighted images have improved the accuracies of clinical diagnosis depending upon magnetic resonance images in stroke as well as inflammations.

  19. New technique for collecting ambient diesel particles for bioassays

    SciTech Connect

    Hallock, M.F.; Smith, T.J.; Hammond, S.K.; Beck, B.D.; Brain, J.D.

    1987-05-01

    This paper describes a new application of viable aerosol sampler, the Liquid electrostatic Aerosol Precipitator (LEAP), for the collection of diesel particles for bioassays of pulmonary toxicity and mutagenicity or carinogenicity. Currently used methods (filtration, dry electrostatic precipitation) cause agglomeration of particles and increases in particle size up to twenty-fold, which may alter particle toxicity significantly. Collection of diesel particles with the LEAP preserved submicronic particle size. Differences in chemical composition of extracts of surface adsorbents as compared to particles collected on filters also were observed. This technique may be applicable for collection other types of combustion products or oil mists that agglomerate when collected by filtration.

  20. A new technique for collecting ambient diesel particles for bioassays.

    PubMed

    Hallock, M F; Smith, T J; Hammond, S K; Beck, B D; Brain, J D

    1987-05-01

    This paper describes a new application of a viable aerosol sampler, the Liquid Electrostatic Aerosol Precipitator (LEAP), for the collection of diesel particles for bioassays of pulmonary toxicity and mutagenicity or carcinogenicity. Currently used methods (filtration, dry electrostatic precipitation) cause agglomeration of particles and increases in particle size up to twenty-fold, which may alter particle toxicity significantly. Collection of diesel particles with the LEAP preserved submicronic particle size. Differences in chemical composition of extracts of surface adsorbents as compared to particles collected on filters also were observed. This technique may be applicable for collection of other types of combustion products or oil mists that agglomerate when collected by filtration. PMID:2438921

  1. Field and Bioassay Indicators for Internal Dose Intervention Therapy

    SciTech Connect

    Carbaugh, Eugene H.

    2007-05-01

    Guidance is presented that is used at the U.S. Department of Energy Hanford Site to identify the potential need for medical intervention in response to intakes of radioactivity. The guidance, based on ICRP Publication 30 models and committed effective dose equivalents of 20 mSv and 200 mSv, is expressed as numerical workplace measurements and derived first-day bioassay results for large intakes. It is used by facility radiation protection staff and on-call dosimetry support staff during the first few days following an intake.

  2. Field and bioassay indicators for internal dose intervention therapy.

    PubMed

    Carbaugh, Eugene H

    2007-05-01

    Guidance is presented that is used at the U.S. Department of Energy Hanford Site to identify the potential need for medical intervention in response to intakes of radioactivity. The guidance, based on ICRP Publication 30 models and committed effective dose equivalents of 20 mSv and 200 mSv, is expressed as numerical workplace measurements and derived first-day bioassay results for large intakes. It is used by facility radiation protection staff and on-call dosimetry support staff during the first few days following an intake. PMID:17440323

  3. Electroantennographic bioassay as a screening tool for host plant volatiles.

    PubMed

    Beck, John J; Light, Douglas M; Gee, Wai S

    2012-01-01

    Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host plant. When a host plant is an important agronomic commodity feeding damage by insect pests can inflict serious economic losses to growers. Accordingly, kairomones can be used as attractants to lure or confuse these insects and, thus, offer an environmentally friendly alternative to pesticides for insect control. Unfortunately, plants can emit a vast number volatiles with varying compositions and ratios of emissions dependent upon the phenology of the commodity or the time of day. This makes identification of biologically active components or blends of volatile components an arduous process. To help identify the bioactive components of host plant volatile emissions we employ the laboratory-based screening bioassay electroantennography (EAG). EAG is an effective tool to evaluate and record electrophysiologically the olfactory responses of an insect via their antennal receptors. The EAG screening process can help reduce the number of volatiles tested to identify promising bioactive components. However, EAG bioassays only provide information about activation of receptors. It does not provide information about the type of insect behavior the compound elicits; which could be as an attractant, repellent or other type of behavioral response. Volatiles eliciting a significant response by EAG, relative to an appropriate positive control, are typically taken on to further testing of behavioral responses of the insect pest. The experimental design presented will detail the methodology employed to screen almond-based host plant volatiles by measurement of the electrophysiological antennal responses of an adult insect pest navel orangeworm (Amyelois transitella) to single components and simple blends of components via EAG bioassay. The method utilizes two excised antennae placed across a "fork" electrode holder. The protocol

  4. An emergency bioassay method for (210)Po in urine.

    PubMed

    Guérin, Nicolas; Dai, Xiongxin

    2015-09-01

    A rapid method was developed to efficiently measure (210)Po in urine samples in an emergency situation. Polonium-210 in small urine samples (10 mL) was spontaneously deposited on a stainless steel disc in 1 M HCl at room temperature for 4 h in a polyethylene bottle. The metallic disc was then counted for 4 h by alpha spectrometry. The developed method allowed the preparation of large sample batch in a short time. The method meets the requirements for an emergency bioassay procedure. PMID:26115206

  5. How to Fabricate Functional Artificial Luciferases for Bioassays.

    PubMed

    Kim, Sung-Bae; Fujii, Rika

    2016-01-01

    The present protocol introduces fabrication of artificial luciferases (ALuc(®)) by extracting the consensus amino acids from the alignment of copepod luciferase sequences. The made ALucs have unique sequential identities that are phylogenetically distinctive from those of any existing copepod luciferase. Some ALucs exhibited heat stability, and strong and greatly prolonged optical intensities. The made ALucs are applicable to various bioassays as an optical readout, including live cell imaging, single-chain probes, and bioluminescent tags of antibodies. The present protocol guides on how to fabricate a unique artificial luciferase with designed optical properties and functionalities. PMID:27424894

  6. 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

  7. Alternative plasticizers for the production of thermo-compressed agar films.

    PubMed

    Sousa, Ana M M; Souza, Hiléia K S; Liu, LinShu; Gonçalves, Maria P

    2015-05-01

    Agar films were produced by thermo-compression using choline chloride (ChCl) as a plasticizer with urea. The three solid components were mixed together with the salt and urea (minor components) added to agar (main component) according to a fixed mass ratio of, respectively, 1.16:1:5. A central composite rotatable design (CCRD) with three parameters, 2(3), was used to evaluate the effects of temperature (X1; °C), time (X2; min) and applied load (X3; kN) of heat-pressing on the maximum tensile strength (TS) of the films (Y; MPa). Mixtures of urea and agar prepared at a mass ratio of 1:5 did not form homogeneous films suggesting the important plasticizing role of the salt. Heat-pressing the mixtures at more draconian conditions led to much darker and opaque films, with better mechanical resistance (higher values of TS). The most resistant film (∼ 15 MPa) was obtained at 140°C, 20 min and 176 kN. Selected films, including the optimal, showed similar water sorption profiles and close values of water vapor permeability (∼ 2.5-3.7 × 10(-9)gm(-1)s(-1)Pa(-1)). The fracture behavior and mechanical properties of the films were greatly affected by additional water plasticization when the films were stored at different conditions of relative humidity. PMID:25727746

  8. Susceptibility testing of Propionibacterium acnes comparing agar dilution with E test.

    PubMed Central

    Smith, M A; Alperstein, P; France, K; Vellozzi, E M; Isenberg, H D

    1996-01-01

    Propionibacterium acnes has been identified as a significant agent of nosocomial infections, including endophthalmitis. Data concerning susceptibility of P. acnes to newer beta-lactam antibiotics and fluoroquinolones are limited. Recent reports suggest that quinolones have activity against these organisms sufficient to warrant further study. We undertook a study to select appropriate antimicrobial agents for use in a rabbit model of P. acnes endophthalmitis. We compared the antibiotic susceptibilities of P. acnes by using the National Committee for Clinical Laboratory Standards method of agar dilution with the E test. Thirteen clinical isolates obtained from eye specimens and three American Type Culture Collection control strains were tested against 14 antibiotics. All the clinical isolates were susceptible by both methods to piperacillin, piperacillin-tazobactam, ampicillin-sulbactam, ticarcillin-clavulanate, cefotaxime, cefotetan, ceftriaxone, cefoxitin, and imipenem in addition to clindamycin but were resistant to metronidazole. The clinical P. acnes isolates also displayed high-level susceptibility to ciprofloxacin, sparfloxacin, and ofloxacin. Almost all the P. acnes strains demonstrated E-test MICs within 2 dilutions of the MICs observed by the agar dilution method. Those few strains for which discrepancies were noted exhibited E-test susceptibilities three- to fivefold dilutions lower than the agar dilution method susceptibilities but only with ampicillin-sulbactam, ticarcillin-clavulanate, and/or clindamycin. On the basis of our study, all of clinical eye isolates were susceptible to these newer antimicrobial agents and the two methods demonstrated similar susceptibility patterns. PMID:8815076

  9. Evaluation of cephamycins as supplements to selective agar for detecting Campylobacter spp. in chicken carcass rinses.

    PubMed

    Chon, Jung-Whan; Kim, Young-Ji; Kim, Hong-Seok; Kim, Dong-Hyeon; Kim, Hyunsook; Song, Kwang-Young; Sung, Kidon; Seo, Kun-Ho

    2016-04-16

    Although cefoperazone is the most commonly used antibiotic in Campylobacter-selective media, the distribution of cefoperazone-resistant bacteria such as extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli is increasing. Here we evaluated the potential of cephamycins for use as supplements to improve modified charcoal-cefoperazone-deoxycholate agar (mCCDA) by replacing cefoperazone with the same concentrations (32 mg/L) of cefotetan (modified charcoal-cefotetan-deoxycholate agar, mCCtDA) and cefoxitin (modified charcoal-cefoxitin-deoxycholate agar, mCCxDA). In chicken carcass rinse samples, the number of mCCDA plates detecting for Campylobacter (18/70, 26%) was significantly lower than that of mCCtDA (42/70, 60%) or mCCxDA plates (40/70, 57%). The number of mCCDA plates (70/70, 100%) that were contaminated with non-Campylobacter species was significantly higher than that of mCCtDA (20/70, 29%) or mCCxDA plates (21/70, 30%). The most common competing species identified using mCCDA was ESBL-producing E. coli, while Pseudomonas species frequently appeared on mCCtDA and mCCxDA. PMID:26915052

  10. Agar Plate Method for Detection and Enumeration of Alkylbenzenesulfonate-Degrading Microorganisms

    PubMed Central

    Ohwada, Kouichi

    1975-01-01

    A simple method for detection and enumeration of alkylbenzenesulfonate (ABS)-degrading microorganisms by using agar plates was developed and used in microbiological studies of coastal marine and polluted river waters. The method depends upon the color responses of neutral red in alkaline medium. Neutral red changes from pink, when it enters into ABS micelles, to yellow, when the ABS is degraded, and does not form micelles. When neutral red-tris(hydroxymethyl)-aminomethane buffer solution and then cationic surfactant solution were sprayed onto the agar surface of ABS-nutrient agar cultures, transparent haloes appeared around the colonies of ABS-degrading microorganisms against a pink background. Viable counts of ABS-degrading bacteria isolated from both seawater and freshwater environments were considerably higher in polluted waters than in less polluted areas. Viable counts of ABS-degrading bacteria averaged 1.5 × 105/ml in samples from the surface water of polluted Tokyo Bay and 3.0 × 104/ml in samples from the surface water of polluted Tamagawa River but were fewer in number in samples from less polluted waters. Images PMID:234155

  11. A supplemented soft agar chemotaxis assay demonstrates the Helicobacter pylori chemotactic response to zinc and nickel

    PubMed Central

    Sanders, Lisa; Andermann, Tessa M.

    2013-01-01

    Directed motility, or chemotaxis, is required for Helicobacter pylori to establish infection in the stomach, although the full repertoire of this bacterium’s chemotactic responses is not yet known. Here we report that H. pylori responds to zinc as an attractant and nickel as a repellent. To reach this conclusion, we employed both a temporal chemotaxis assay based on bacterial reversals and a supplemented soft agar spatial assay. We refined the temporal assay using a previously described chemorepellent, acid, and found that H. pylori requires rich media with serum to maintain optimal swimming motility. Surprisingly, we found that some strains respond to acid as an attractant, and that the TlpC chemoreceptor correlated with whether acid was sensed as an attractant or repellent. Using this same assay, we detected weak repellent responses to nickel and copper, and a varied response to zinc. We thus developed an alternative spatial chemotactic assay called the supplemented soft agar assay, which utilizes soft agar medium supplemented with the test compound. With Escherichia coli, the attractant serine slowed overall bacterial migration, while the repellent nickel increased the speed of overall migration. In H. pylori we detected slowed migration with doubled tryptone media, as well as zinc, consistent with an attractant response. In contrast, nickel increased migration, consistent with repulsion. PMID:23139399

  12. 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

  13. 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

  14. Development and characterisation of an agar--polyvinyl alcohol blend hydrogel.

    PubMed

    Lyons, John G; Geever, Luke M; Nugent, Michael J D; Kennedy, James E; Higginbotham, Clement L

    2009-10-01

    Numerous authors have reported on hydrogel technologies providing products suitable for applications in biomedical, personal care as well as in nano-sensor applications. Hydrogels fabricated from single polymers have been extensively investigated. However, in many cases a single polymer alone cannot meet divergent demands in terms of both properties and performance. In this work, hydrogels were prepared by physically blending the natural polymer agar with polyvinyl alcohol in varying ratios to produce a new biosynthetic polymer applicable for a variety of purposes. Hydrogen bonding was observed to take place between the polyvinyl alcohol and the agar molecules in the composite materials leading to changes in the thermal, mechanical and swelling characteristics of the composite hydrogels. The composite hydrogels exhibited a slightly higher melting temperature than pure agar (116.81 degrees C). Irreversible compressive damage was found to occur at lower strain levels during compression testing of the dehydrated samples consisting of higher PVOH concentrations. Rheological analysis of hydrated sample revealed G' values of between 5000 and 10,000 Pa for the composite blends, with gels containing higher PVOH percentages exhibiting poorer mechanical strength. PMID:19627855

  15. Isolation of Clostridium difficile from faecal specimens--a comparison of chromID C. difficile agar and cycloserine-cefoxitin-fructose agar.

    PubMed

    Carson, Kerry C; Boseiwaqa, Lusiana V; Thean, Sara K; Foster, Niki F; Riley, Thomas V

    2013-09-01

    The culture of toxigenic Clostridium difficile from stool specimens is still seen as the gold standard for the laboratory diagnosis of C. difficile infection (CDI). bioMérieux have released ChromID Cdiff chromogenic agar (CDIF) for the isolation and identification of C. difficile in 24 h. In this study, we compared CDIF to pre-reduced cycloserine-cefoxitin-fructose agar with sodium taurocholate (TCCFA) in the examination of glutamate dehydrogenase-positive faecal specimens that were either GeneOhm positive or negative, using direct culture or culture following alcohol shock. Direct culture on CDIF had a sensitivity of 100 % and recovery of 94 % while for TCCFA these were 87 % and 82 %, respectively. For GeneOhm-positive alcohol-shocked faecal samples, sensitivity and recovery on CDIF was similar to direct culture while on TCCFA they were about 10 % higher. For direct culture, there was a significant difference between growth on CDIF at 24 h and TCCFA at 48 h (P = 0.001) and between the two media at 48 h (P<0.001). A total of 142 strains of C. difficile were recovered in pure culture from all GeneOhm-positive samples used in this study and 11 (7.7 %) of these were A(-)B(-)CDT(-) and may represent mixed infections of toxigenic and non-toxigenic C. difficile. The most dominant ribotype was UK 014 (14.7 %) followed by 002 (11.9 %) and 020 (11.9 %), and 36 % of toxigenic isolates, including an A(-)B(+)CDT(-) strain, could not be assigned a UK ribotype. CDIF outperformed pre-reduced TCCFA by negating the need for alcohol shock treatment and by giving a time saving of 24 h in the isolation of C. difficile. CDIF plates were also more selective than TCCFA and C. difficile colonies were easy to identify and subculture prior to strain typing. PMID:23579394

  16. 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

  17. Agar disk elution method for susceptibility testing of Mycobacterium marinum and Mycobacterium fortuitum complex to sulfonamides and antibiotics.

    PubMed Central

    Stone, M S; Wallace, R J; Swenson, J M; Thornsberry, C; Christensen, L A

    1983-01-01

    An agar disk elution method using round well plates, supplemented Mueller-Hinton agar, and commercial drug disks is described for susceptibility testing of Mycobacterium marinum and the rapidly growing mycobacteria to antibiotics and sulfonamides. By this method, 14 of 14 strains of M. marinum were susceptible to rifampin, doxycycline, minocycline, and trimethoprim-sulfamethoxazole. Identical results were obtained with Middlebrook 7H10 agar and drugs prepared from standard powders. With 58 isolates of Mycobacterium fortuitum and Mycobacterium chelonei, this method had a 92% correlation with broth minimal inhibitory concentration determinations for cefoxitin and greater than 98% for doxycycline, kanamycin, amikacin, and the sulfonamides. Sixty-nine percent of isolates of M. chelonei susceptible to amikacin on supplemented Mueller-Hinton agar were resistant on 7H10 agar, and 15 of 16 M. chelonei isolates susceptible to erythromycin in broth were resistant by disk elution when an endpoint of no growth was used with either agar. The agar disk elution method offers a practical method for testing of most antibacterial agents against these mycobacterial species. Images PMID:6651277

  18. Chemical Exacerbation of Light-induced Retinal Degeneration in F344/N Rats in National Toxicology Program Rodent Bioassays.

    PubMed

    Yamashita, Haruhiro; Hoenerhoff, Mark J; Peddada, Shyamal D; Sills, Robert C; Pandiri, Arun R

    2016-08-01

    Retinal degeneration due to chronic ambient light exposure is a common spontaneous age-related finding in albino rats, but it can also be related to exposures associated with environmental chemicals and drugs. Typically, light-induced retinal degeneration has a central/hemispherical localization whereas chemical-induced retinal degeneration has a diffuse localization. This study was conducted to identify and characterize treatment-related retinal degeneration in National Toxicology Program rodent bioassays. A total of 3 chronic bioassays in F344/N rats (but not in B6C3F1/N mice) were identified that had treatment-related increases in retinal degeneration (kava kava extract, acrylamide, and leucomalachite green). A retrospective light microscopic evaluation of the retinas from rats in these 3 studies showed a dose-related increase in the frequencies of retinal degeneration, beginning with the loss of photoreceptor cells, followed by the inner nuclear layer cells. These dose-related increased frequencies of degenerative retinal lesions localized within the central/hemispherical region are suggestive of exacerbation of light-induced retinal degeneration. PMID:27230502

  19. [Evaluation of Antilles fish ciguatoxicity by mouse and chick bioassays].

    PubMed

    Pottier, I; Vernoux, J P

    2003-03-01

    Ciguatera is a common seafood poisoning in Western Atlantic and French West Indies. Ciguatera fish poisoning in the Caribbean is a public health problem. A toxicological study was carried out on 178 Caribbean fish specimens (26 species) captured off Guadeloupe and Saint Barthelemy between 1993 and 1999. The mouse bioassay and the chick feeding test were used to control fish edibility. Ciguatoxins presence was assumed when symptomatology was typical of ciguatera in mouse and chick. Fishes were classified in three groups: non toxic fish (edible), low toxic fish (not edible) and toxic fish (not edible). 75% of fishes were non toxic. Toxic fish specimens belonged to four families of high trophic level carnivores: Carangidae, Lutjanidae, Serranidae et Sphyraenidae. Percentages of toxic fishes to humans reached 55% for Caranx latus and 33% for Caranx bartholomaei and Caranx lugubris. Only a significant correlation between weight and toxicity was only found for C. latus and snappers. Small carnivorous groupers (Serranidae) were also toxic. Atoxic fish species were (a) pelagic fish (Coryphaena hippurus, Auxis thazard and Euthynnus pelamis), (b) invertebrates feeders (Malacanthus plumieri, Balistes vetula), (c) small high-risk fish or (d) fish of edible benthic fish families. Liver of four fishes (Mycteroperca venenosa, Caranx bartholomaei, Seriola rivoliana, Gymnothorax funebris) contained ciguatoxins at a significant level although their flesh was safe. This study confirms the usefulness of mouse and chick bioassays for sanitary control of fish. PMID:12784589

  20. A Bioassay for Lafora Disease and Laforin Glucan Phosphatase Activity

    PubMed Central

    Sherwood, Amanda R.; Johnson, Mary Beth; Delgado-Escueta, Antonio V.; Gentry, Matthew S.

    2013-01-01

    Objectives Lafora disease is a rare yet invariably fatal form of progressive neurodegenerative epilepsy resulting from mutations in the phosphatase laforin. Several therapeutic options for Lafora disease patients are currently being explored, and these therapies would benefit from a biochemical means of assessing functional laforin activity following treatment. To date, only clinical outcomes such as decreases in seizure frequency and severity have been used to indicate success of epilepsy treatment. However, these qualitative measures exhibit variability and must be assessed over long periods of time. In this work, we detail a simple and sensitive bioassay that can be used for the detection of functional endogenous laforin from human and mouse tissue. Design and methods We generated antibodies capable of detecting and immunoprecipitating endogenous laforin. Following laforin immunoprecipitation, laforin activity was assessed via phosphatase assays using para-nitrophenylphosphate (pNPP) and a malachite green-based assay specific for glucan phosphatase activity. Results We found that antibody binding to laforin does not impede laforin activity. Furthermore, the malachite green-based glucan phosphatase assay used in conjunction with a rabbit polyclonal laforin antibody was capable of detecting endogenous laforin activity from human and mouse tissue. Importantly, this assay discriminated between laforin activity and other phosphatases. Conclusions The bioassay that we have developed utilizing laforin antibodies and an assay specific for glucan phosphatase activity could prove valuable in the rapid detection of functional laforin in patients to which novel Lafora disease therapies have been administered. PMID:24012855

  1. Dichloromethane attracts diabroticite larvae in a laboratory behavioral bioassay.

    PubMed

    Jewett, D K; Bjostad, L B

    1996-07-01

    A two-choice laboratory behavioral bioassay was used to demonstrate that dichloromethane elicits the dose-dependent attraction of secondinstar western and southern corn rootworms. Preliminary data suggest that second-instar banded cucumber beetles are also attracted to dichloromethane. An eluotropic series of 10 materials, including distilled water, ethanol, methanol, acetone, ethyl dichloroacetate, dichloromethane, diethyl ether, benzene, hexadecane, and hexane, was tested for attraction of western corn rootworm larvae. Dichloromethane was the only one attractive at all doses tested, and orthogonal comparisons revealed a quadratic trend (convex) for responses of larvae to increasing dose. Benzene and hexadecane also attracted larvae, but significantly fewer than dichloromethane, and only at three doses and one dose, respectively. Orthogonal comparisons revealed no linear or quadratic trend for responses of larvae to increasing doses of either compound. Dichloromethane is the first organic compound demonstrated to attract western corn rootworm larvae in the absence of carbon dioxide. Carbon dioxide has previously been reported to attract western corn rootworm larvae either independently or when combined with other organic compounds, and the sensitivity of our bioassay was tested by demonstrating the dose-dependent attraction of western corn rootworm larvae to carbonated water as a carbon dioxide source. We have also demonstrated the attraction of southern corn rootworm larvae to carbon dioxide and propose that carbon dioxide and dichloromethane behave analogously when they interact with chemoreceptor sites on larvae. PMID:24226089

  2. Vicia faba bioassay for environmental toxicity monitoring: A review.

    PubMed

    Iqbal, Munawar

    2016-02-01

    Higher plants are recognized as excellent genetic models to detect cytogenetic and mutagenic agents and are frequently used in environmental monitoring studies. Vicia faba (V. faba) bioassay have been used to study DNA damages i.e., chromosomal and nuclear aberrations induced by metallic compounds, pesticides, complex mixtures, petroleum derivates, toxins, nanoparticles and industrial effluents. The main advantages of using V. faba is its availability round the year, economical to use, easy to grow and handle; its use does not require sterile conditions, rate of cell division is fast, chromosomes are easy to score, less expensive and more sensitive as compared to other short-term tests that require pre-preparations. The V. faba test offers evaluation of different endpoints and tested agents can be classified as cytotoxic/genotoxic/mutagenic. This test also provides understanding about mechanism of action, whether the tested agent is clastogenic or aneugenic in nature. In view of advantages offered by V. faba test system, it is used extensively to assess toxic agents and has been emerged as an important bioassay for ecotoxicological studies. Based on the applications of V. faba test to assess the environmental quality, this article offers an overview of this test system and its efficiency in assessing the cytogenetic and mutagenic agents in different classes of the environmental concerns. PMID:26414739

  3. 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

  4. 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

  5. Resistance monitoring of Heliothis virescens to pyramided cotton varieties with a hydrateable, artificial cotton leaf bioassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proof of concept was demonstrated for a practical, off the shelf bioassay to monitor for tobacco budworm resistance to pyramided Bt cotton using plant eluants. The bioassay was based on a previously described feeding disruption test using hydrateable artificial diet containing a blue indicator dye, ...

  6. Comparison of two mosquito bioassay methods for the estimate of minimum effective dose in repellents

    Technology Transfer Automated Retrieval System (TEKTRAN)

    It is expected that laboratory-based repellent bioassays should reliably evaluate the efficacy of compounds that deter mosquito feeding behavior. The variety of repellent bioassays available allows for flexibility in design, but makes it difficult to compare any two methods, including in vitro and i...

  7. Profiling Animal Toxicants by Automatically Mining Public Bioassay Data: A Big Data Approach for Computational Toxicology

    PubMed Central

    Zhang, Jun; Hsieh, Jui-Hua; Zhu, Hao

    2014-01-01

    In vitro bioassays have been developed and are currently being evaluated as potential alternatives to traditional animal toxicity models. Already, the progress of high throughput screening techniques has resulted in an enormous amount of publicly available bioassay data having been generated for a large collection of compounds. When a compound is tested using a collection of various bioassays, all the testing results can be considered as providing a unique bio-profile for this compound, which records the responses induced when the compound interacts with different cellular systems or biological targets. Profiling compounds of environmental or pharmaceutical interest using useful toxicity bioassay data is a promising method to study complex animal toxicity. In this study, we developed an automatic virtual profiling tool to evaluate potential animal toxicants. First, we automatically acquired all PubChem bioassay data for a set of 4,841 compounds with publicly available rat acute toxicity results. Next, we developed a scoring system to evaluate the relevance between these extracted bioassays and animal acute toxicity. Finally, the top ranked bioassays were selected to profile the compounds of interest. The resulting response profiles proved to be useful to prioritize untested compounds for their animal toxicity potentials and form a potential in vitro toxicity testing panel. The protocol developed in this study could be combined with structure-activity approaches and used to explore additional publicly available bioassay datasets for modeling a broader range of animal toxicities. PMID:24950175

  8. Effects of Wind Speed on Aerosol Spray Penetration in Adult Mosquito Bioassay Cages

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioassay cages are commonly used to assess efficacy of insecticides against adult mosquitoes in the field. To properly correlate adult mortality readings to insecticidal efficacy and/or spray application parameters, it is important to know how the cage used in the bioassay interacts with the spray ...

  9. LIFE CYCLE BIOASSAY FOR ASSESSMENT OF THE EFFECTS OF TOXIC CHEMICALS USING RAPID CYCLING OF BRASSICA

    EPA Science Inventory

    Initial evaluation of a new plant life cycle bioassay for the assessment of the effects of toxic chemicals is presented. he bioassay features a rapid cycling Brassica species that can complete its life cycle in as little as 36 days. he herbicide dalapon (2,2 dichloropropionic aci...

  10. Immunochemical technologies for replacement of rodent bioassays in sensitive detection of toxins in foods

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Rapid sensitive assays for biothreat toxins that can be used to detect intentionally contaminated foods are now typically performed via bioassay in live mice. While bioassay provides essential data on bioavailability, animal models are technically, fiscally, and ethically challenging. Through carefu...

  11. Improved high-throughput bioassay for Rhyzopertha dominica (F.) (Coleoptera: Bostrichidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    As we gain more information through functional genomic studies of Rhyzopertha dominica (F.), we need a high throughput bioassay system to screen potential biopesticides. R. dominica is an internal feeder during immature stages and presents unique challenges with traditional bioassay methods. Our pri...

  12. Development of a High Throughput Translational Bioassay for Plant Biofuel Properties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using the well developed microbial system, Clostridium phytofermentans, we have developed a robust bioassay for biomass digestibility and conversion to biofuels. The bioassay can be used to measure the impact of plant genetic diversity on digestibility, and thereby determine the potential effects of...

  13. A Bioassay for Determining Resistance Levels in Tarnished Plant Bug Populations to Neonicotinoid Insecticides

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A laboratory bioassay was developed and used to test field populations of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), for resistance development to the neonicitinoid insecticides imidacloprid (Trimax®) and thiamethoxam (Centric®). The bioassay determined LC50 values by feeding...

  14. A LABORATORY BIOASSAY FOR MONITORING RESISTANCE IN TARNISHED PLANT BUG POPULATIONS TO NEONICOTINOID INSECTICIDES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A laboratory bioassay was developed for testing tarnished plant bug populations for resistance development to the neonicotinoid insecticides imidacloprid and thiamethoxam. The bioassay allows for the determination of LC50 values by feeding known doses of the insecticides to adult tarnished plant bu...

  15. ACINETOBACTER SPP.: DISTINCT MORPHOLOGY ON EOSIN METHYLENE BLUE AGAR AS AN AID TO IDENTIFICATION IN DRINKING WATER

    EPA Science Inventory

    'Acinetobacter calcoaceticus', frequently found in drinking waters and implicated in nosocomial infections, was presumptively identified by its tiny, blue colonial appearance on Levine eosin methylene blue agar. All of the 33 isolates from drinking water showing this distinctive ...

  16. Benthic invertebrate bioassays with toxic sediment and pore water

    USGS Publications Warehouse

    Giesy, John P.; Rosiu, Cornell J.; Graney, Robert L.; Henry, Mary G.

    1990-01-01

    The relative sensitivities of bioassays to determine the toxicity of sediments were investigated and three methods of making the sample dilutions required to generate dose-response relationships were compared. The assays studied were: (a) Microtox®, a 15-min assay ofPhotobacterium phosphoreum bioluminescence inhibition by pore water; (b) 48-h Daphnia magnalethality test in pore water; (c) 10-d subchronic assay of lethality to and reduction of weight gain by Chironomus tentans performed in either whole sediment or pore water; (d) 168-h acute lethality assay of Hexagenia limbata in either whole sediment or pore water. The three methods of diluting sediments were: (a) extracting pore water from the toxic location and dilution with pore water from the control station; (b) diluting whole sediment from the toxic location with control whole sediment from a reference location, then extracting pore water; and (c) diluting toxic, whole sediment with whole sediment from a reference location, then using the whole sediment in bioassays. Based on lethality, H. limbata was the most sensitive organism to the toxicity of Detroit River sediment. Lethality of D. magna in pore water was similar to that of H. limbata in whole sediment and can be used to predict effects of whole sediment toxicity to H. limbata. The concentration required to cause a 50% reduction in C. tentans growth (10-d EC50) was approximately that which caused 50% lethality of D. magna (48-h LC50) and was similar to the toxicity that restricts benthic invertebrate colonization of contaminated sediments. While the three dilution techniques gave similar results with some assays, they gave very different results in other assays. The dose-response relationships determined by the three dilution techniques would be expected to vary with sediment, toxicant and bioassay type, and the dose-response relationship derived from each technique needs to be interpreted accordingly.

  17. Evaluation of the mutagenicity and carcinogenicity of motor vehicle emissions in short-term bioassays.

    PubMed Central

    Lewtas, J

    1983-01-01

    Incomplete combustion of fuel in motor vehicles results in the emission of submicron carbonaceous particles which, after cooling and dilution, contain varying quantities of extractable organic constituents. These organics are mutagenic in bacteria. Confirmatory bioassays in mammalian cells provide the capability of detecting chromosomal and DNA damage in addition to gene mutations. In order to evaluate the mutagenicity of these organics in mammalian cells, extractable organics from particle emissions from several diesel and gasoline vehicles were compared in a battery of microbial, mammalian cell and in vivo bioassays. The mammalian cell mutagenicity bioassays were selected to detect gene mutations, DNA damage, and chromosomal effects. Carcinogenesis bioassays conducted included short-term assays for oncogenic transformation and skin tumorigenesis. The results in different assay systems are compared both qualitatively and quantitatively. Good quantitative correlations were observed between several mutagenesis and carcinogenesis bioassays for this series of diesel and gasoline emissions. PMID:6186475

  18. In vitro/in vivo evaluation of agar nanospheres for pulmonary delivery of bupropion HCl.

    PubMed

    Varshosaz, Jaleh; Minaiyan, Mohsen; Zaki, Mohammad Reza; Fathi, Milad; Jaleh, Hossein

    2016-07-01

    Bupropion HCl is an atypical antidepressant drug with rapid and high first-pass metabolism. Sustained release dosage form of this drug is suggested for reducing its side effects which are mainly seizures. The aim of the present study was to design pulmonary agar nanospheres of bupropion HCl with effective systemic absorption and extended release properties. Bupropion HCl was encapsulated in agar nanospheres by ionic gelation, and characterized for physical and release properties. Pharmacokinetic studies on nanospheres were performed on rats by intratracheal spraying of 5 mg/kg of drug in form of nanospheres compared to intravenous and pulmonary delivery of the same dose as simple solution of the drug. The optimized nanoparticles showed particle size of 320 ± 90 nm with polydispersity index of 0.85, the zeta potential of -29.6 mV, drug loading efficiency of 43.1 ± 0.28% and release efficiency of 66.7 ± 2%. The area under the serum concentration-time profile for the pulmonary nanospheres versus simple solution was 10 237.84 versus 28.8 µg/ml min, Tmax of 360 versus 60 min and the Cmax of 1927.93 versus9.93 ng/ml, respectively. The absolute bioavailability of the drug was 86.69% for nanospheres and 0.25% for pulmonary simple solution. Our results indicate that pulmonary delivery of bupropion loaded agar nanospheres achieves systemic exposure and extends serum levels of the drug. PMID:25835223

  19. Cavitation-enhanced delivery of insulin in agar and porcine models of human skin

    NASA Astrophysics Data System (ADS)

    Feiszthuber, Helga; Bhatnagar, Sunali; Gyöngy, Miklós; Coussios, Constantin-C.

    2015-03-01

    Ultrasound-assisted transdermal insulin delivery offers a less painful and less invasive alternative to subcutaneous insulin injections. However, ultrasound-based drug delivery, otherwise known as sonophoresis, is a highly variable phenomenon, in part dependent on cavitation. The aim of the current work is to investigate the role of cavitation in transdermal insulin delivery. Fluorescently stained, soluble Actrapid insulin was placed on the surface of human skin-mimicking materials subjected to 265 kHz, 10% duty cycle focused ultrasound. A confocally and coaxially aligned 5 MHz broadband ultrasound transducer was used to detect cavitation. Two different skin models were used. The first model, 3% agar hydrogel, was insonated with a range of pressures (0.25-1.40 MPa peak rarefactional focal pressure—PRFP), with and without cavitation nuclei embedded within the agar at a concentration of 0.05% w/v. The second, porcine skin was insonated at 1.00 and 1.40 MPa PRFP. In both models, fluorescence measurements were used to determine penetration depth and concentration of delivered insulin. Results show that in agar gel, both insulin penetration depth and concentration only increased significantly in the presence of inertial cavitation, with up to a 40% enhancement. In porcine skin the amount of fluorescent insulin was higher in the epidermis of those samples that were exposed to ultrasound compared to the control samples, but there was no significant increase in penetration distance. The results underline the importance of instigating and monitoring inertial cavitation during transdermal insulin delivery.

  20. Cavitation-enhanced delivery of insulin in agar and porcine models of human skin.

    PubMed

    Feiszthuber, Helga; Bhatnagar, Sunali; Gyöngy, Miklós; Coussios, Constantin-C

    2015-03-21

    Ultrasound-assisted transdermal insulin delivery offers a less painful and less invasive alternative to subcutaneous insulin injections. However, ultrasound-based drug delivery, otherwise known as sonophoresis, is a highly variable phenomenon, in part dependent on cavitation. The aim of the current work is to investigate the role of cavitation in transdermal insulin delivery. Fluorescently stained, soluble Actrapid insulin was placed on the surface of human skin-mimicking materials subjected to 265 kHz, 10% duty cycle focused ultrasound. A confocally and coaxially aligned 5 MHz broadband ultrasound transducer was used to detect cavitation. Two different skin models were used. The first model, 3% agar hydrogel, was insonated with a range of pressures (0.25-1.40 MPa peak rarefactional focal pressure-PRFP), with and without cavitation nuclei embedded within the agar at a concentration of 0.05% w/v. The second, porcine skin was insonated at 1.00 and 1.40 MPa PRFP. In both models, fluorescence measurements were used to determine penetration depth and concentration of delivered insulin. Results show that in agar gel, both insulin penetration depth and concentration only increased significantly in the presence of inertial cavitation, with up to a 40% enhancement. In porcine skin the amount of fluorescent insulin was higher in the epidermis of those samples that were exposed to ultrasound compared to the control samples, but there was no significant increase in penetration distance. The results underline the importance of instigating and monitoring inertial cavitation during transdermal insulin delivery. PMID:25716689

  1. [Change of leukocytic phagocytosis during repeat hemoperfusion with cross-linked agar beads entrapped attapulgite clay].

    PubMed

    Huang, Wei; Ma, Yu; Yang, Xiaolan; Tang, Xianjue; Shu, Changda

    2003-06-01

    The leukocytic phagocytosis rate and the index of phagocytosis of rats on cross-linked agar beads entrapped attapulgite clay (CAA) hemoperfusion were studied. The results revealed that the leukocytic phagocytosis rate and the index of phagocytosis descended significantly after 1 hour and rose gradually after 6 hours. Finally it reached the normal level after 48 hours. Hemoperfusion repeated two times gave similar results. In conclusion, the function of leukocytic phagocytosis declined temporarily during CAA hemoperfusion. Many times hemoperfusion will not notably affect the body's defense system of rats. PMID:12856604

  2. Coma in the course of severe poisoning after consumption of red fly agaric (Amanita muscaria).

    PubMed

    Mikaszewska-Sokolewicz, Małgorzata A; Pankowska, Sylwestra; Janiak, Marek; Pruszczyk, Piotr; Łazowski, Tomasz; Jankowski, Krzysztof

    2016-01-01

    Red fly agaric poisoning is rare. It can be consumed for suicidal purposes or its psychedelic effect. The paper describes the case of a young men, who fell into a coma after ingestion of the red toadstools. Quick identification of the poison, early use of gastric lavage and symptomatic treatment resulted in regression of symptoms and lead to the patient's discharge from the hospital on the third day after intoxication. Authors discussing the poisonous alkaloids contained in the red toadtools: ibotenic acid, muscimol, muscasone and muscarine and theirs properties, responsible for the symptoms of intoxication. PMID:26828668

  3. CHROMagar Yersinia, a New Chromogenic Agar for Screening of Potentially Pathogenic Yersinia enterocolitica Isolates in Stools

    PubMed Central

    Renaud, Nicolas; Lecci, Laetitia; Courcol, René J.; Simonet, Michel

    2013-01-01

    CHROMagar Yersinia (CAY) is a new chromogenic medium for the presumptive detection of virulent Yersinia enterocolitica in stools. Based on a comparative analysis of 1,494 consecutive stools from hospitalized patients, CAY was found to be just as sensitive as the reference medium (cefsulodin-irgasan-novobiocin agar) but was significantly more specific and had a very low false-positive rate. CAY reduces the workload (and thus costs) for stool analysis and can therefore be recommended for routine laboratory use. PMID:23363840

  4. Comparative study of 6-APA production by free and agar immobilized bacteria in nutrient broth culture.

    PubMed

    Dolui, A K; Das, S

    2011-04-01

    In the present study different bacterial samples were isolated from soil of different places of Dibrugarh and screened for biotransformation ability to produce 6-Aminopenicillanic acid. Among ten isolated bacterial samples, three gram positive bacterial samples designated as AKDD-2, AKDD-4 and AKDD-6 showed the production of 6-APA from penicillin G. Assessment of production of 6-APA after incubation in penicillin G (2 mg/ml) by three different samples separately in free and agar immobilization state was done by HPLC analysis. Reusability of immobilized cells was found successful up to 14 days. PMID:21614893

  5. Amino acid mediated synthesis of silver nanoparticles and preparation of antimicrobial agar/silver nanoparticles composite films.

    PubMed

    Shankar, Shiv; Rhim, Jong-Whan

    2015-10-01

    Silver nanoparticles (AgNPs) were synthesized using amino acids (tyrosine and tryptophan) as reducing and capping agents, and they were incorporated into the agar to prepare antimicrobial composite films. The AgNPs solutions exhibited characteristic absorption peak at 420 nm that showed a red shift to ∼434 nm after forming composite with agar. XRD data demonstrated the crystalline structure of AgNPs with dominant (111) facet. Apparent surface color and transmittance of agar films were greatly influenced by the AgNPs. The incorporation of AgNPs into agar did not exhibit any change in chemical structure, thermal stability, moisture content, and water vapor permeability. The water contact angle, tensile strength, and modulus decreased slightly, but elongation at break increased after AgNPs incorporation. The agar/AgNPs nanocomposite films possessed strong antibacterial activity against Listeria monocytogenes and Escherichia coli. The agar/AgNPs film could be applied to the active food packaging by controlling the food-borne pathogens. PMID:26076636

  6. Chronic and Initiation/Promotion Skin Bioassays of Petroleum Refinery Streams.

    PubMed Central

    Skisak, C; Furedi-Machacek, EM; Schmitt, SS; Swanson, MS; Vernot, EH

    1994-01-01

    Nine refinery streams were tested in both chronic and initiation/promotion (I/P) skin bioassays. In the chronic bioassay, groups of 50 C3H/HeJ mice received twice weekly applications of 50 microl of test article for at least 2 years. In the initiation phase of the I/P bioassay, groups of CD-1 mice received an initiating dose of 50 microl of test article for 5 consecutive days, followed by promotion with 50 microl of phorbol-12-myristate-13-acetate (0.01% w/v in acetone) for 25 weeks. In the promotion phase of the I/P bioassay, CD-1 mice were initiated with 50 microl of 7,12-dimethylbenzanthracene (0.1% w/v in acetone) or acetone, followed by promotion with 50 microl of test article twice weekly for 25 weeks. The most volatile of the streams, sweetened naphtha, and the least volatile, vacuum residuum, were noncarcinogenic in both assays. Middle distillates, with a boiling range of 150 degrees-370 degreesC, demonstrated carcinogenic activity in the chronic bioassay and acted as promoters but not initiators in the I/P bioassay. Untreated mineral oil streams displayed initiating activity and were carcinogenic in the chronic bioassay, presumably due to the presence of polycyclic aromatic hydrocarbons of requisite size and structure. A highly solvent-refined mineral oil stream lacked initiating activity. These results indicate that the I/P bioassay, which takes 6 months to complete, may be a good qualitative predictor of the results of a chronic bioassay, at least for petroleum streams. Furthermore, the I/P bioassay can provide insight into possible mechanisms of tumor development. Images p82-a PMID:9719673

  7. Harvester ant bioassay for assessing hazardous chemical waste sites

    SciTech Connect

    Gano, K.A.; Carlile, D.W.; Rogers, L.E.

    1984-12-01

    A technique was developed for using harvester ants, Pogonomyrmex owhyeei, in terrestrial bioassays. Procedures were developed for maintaining stock populations, handling ants, and exposing ants to toxic materials. Relative toxicities were determined by exposing ants to 10 different materials. These materials included three insecticides, Endrin, Aldrin, and Dieldrin; one herbicide, 2,4-D; three oil-like compounds, wood preservative, drilling fluid, and slop oil; and three heavy metals, copper, zinc, and cadmium. Ants were exposed in petri dishes containing soil amended with a particular toxicant. Under these test conditions, ants showed no sensitivity to the metals or 2,4-D. Ants were sensitive to the insecticides and oils in repeated tests, and relative toxicity remained consistent throughout. Aldrin was the most toxic material, followed by Dieldrin, Endrin, wood preservative, drilling fluid, and slop oil. 10 refs., 2 figs., 2 tabs.

  8. Acute bioassays with benthic macroinvertebrates conducted in situ

    SciTech Connect

    Whaley, M.; Garcia, R.; Sy, J. )

    1989-10-01

    Several methods of toxicity testing using macroinvertebrates in controlled laboratory experiments have been reported. Researchers conducted bioassays with natural assemblages of benthic macroinvertebrates exposed to several petroleum refinery effluents. They found that the populations of invertebrates declined after only a few days of exposure. The objective of the study was to determine the acute toxic effects of discharge water from a petrochemical complex on a natural assemblage of benthic macroinvertebrates. The discharge water consisted of refinery wastewater and sanitary wastewater, as well as brine discharge from a power/desalination plant. The benthic macroinvertebrates were transplanted from a healthy reef area to the outfall channel receiving the discharge water. The study began on October 7, 1985, and concluded that same week. Any decrease in specific species would indicate that the discharge was toxic to these species. These species could also serve as indicators of toxic conditions at other locations.

  9. A sediment suspension system for bioassays with small aquatic organisms

    USGS Publications Warehouse

    Schmidt-Dallmier, M. J.; Atchison, G.J.; Steingraeber, M.T.; Knights, B.C.

    1992-01-01

    Exposure of aquatic organisms to suspended sediments can impair growth and survival and increase bioaccumulation of sediment-associated contaminants. However, evaluation of the effects of suspended sediments and their associated contaminants on aquatic organisms has been hampered by the lack of a practical and inexpensive exposure system for conducting bioassays. We present a cost-effective system for assessing the effects of suspended sediments and associated contaminants on small aquatic organisms. A 7-day suspension test was conducted with nominal sediment concentrations ranging from 0.0 To 5.0 g 1-1. The system maintained relatively constant suspended sediment concentrations, as measured by turbidity, and caused minimal mortality to test organisms.

  10. Use of bioassay methods to evaluate incinerator emissions

    SciTech Connect

    Watts, R.R.; DeMarini, D.M.; Linak, W.P.; Lemieux, P.M.; McSorley, J.A.

    1989-01-01

    The organic components in combustion emissions are composed of thousands of chemicals. Analyzing such a complex mixture for the presence of even a few selected chemicals is difficult and provides information on only a fraction of the chemicals present. Reliance on such limited chemical analysis for determining possible health effects may ignore the contribution of many other chemical components of the effluent. Because combustion emissions are complex mixtures, they have been evaluated as such, rather than by studying a few selected chemicals that might be present. The Salmonella (Ames) assay was used to determine the mutagenicity associated with particles from the effluent of municipal-waste combustors, from ambient air collected near a municipal-waste combustor, and from the effluent of a pilot-sized rotary kiln in which polyethylene was combusted. Filter samples were extracted with dichloromethane, and concentrated extracts were solvent exchanged into dimethyl sulfoxide for bioassay.

  11. Toxicity assessment using different bioassays and microbial biosensors.

    PubMed

    Hassan, Sedky H A; Van Ginkel, Steven W; Hussein, Mohamed A M; Abskharon, Romany; Oh, Sang-Eun

    2016-01-01

    Toxicity assessment of water streams, wastewater, and contaminated sediments, is a very important part of environmental pollution monitoring. Evaluation of biological effects using a rapid, sensitive and cost effective method can indicate specific information on ecotoxicity assessment. Recently, different biological assays for toxicity assessment based on higher and lower organisms such as fish, invertebrates, plants and algal cells, and microbial bioassays have been used. This review focuses on microbial biosensors as an analytical device for environmental, food, and biomedical applications. Different techniques which are commonly used in microbial biosensing include amperometry, potentiometry, conductometry, voltammetry, microbial fuel cells, fluorescence, bioluminescence, and colorimetry. Examples of the use of different microbial biosensors in assessing a variety of environments are summarized. PMID:27071051

  12. Synthesis, bioassay, crystal structure and ab initio studies of Erlenmeyer azlactones

    NASA Astrophysics Data System (ADS)

    Parveen, Mehtab; Ali, Akhtar; Ahmed, Sarfaraz; Malla, Ali Mohammed; Alam, Mahboob; Pereira Silva, P. S.; Silva, Manuela Ramos; Lee, Dong-Ung

    2013-03-01

    Several 4-arylidene-2-phenyl-5(4H)-azlactones have been synthesized via Erlenmeyer method. The synthesized compounds have been characterized on the basis of systematic spectral studies (IR, 1H NMR, 13C NMR, and MS). The compound (4Z)-4-(3,5-dimethoxybenzylidene)-2-phenyl-1,3-oxazol-5(4H)-one, C18H15NO4, (5), crystallizes in the orthorhombic system, space group P212121, with a = 5.6793(3) Å, b = 15.2038(7) Å, c = 17.6919(10) Å, Mr = 309.31, V = 1527.64(14) Å3, Z = 4 and R = 0.0547. The compound (4Z)-2-phenyl-4-(3,4,5-trimethoxybenzylidene)-1,3-oxazol-5(4H)-one, C19H17NO5, (6) crystallizes in triclinic geometry with space group P-1, having unit cell parameters a = 7.3814(3) Å, b = 8.1446(3) Å, c = 13.9845(5) Å, α = 86.918(3), β = 83.314(2), γ = 82.462(3), Mr = 339.34, V = 827.16(5) Å3, Z = 2 and R = 0.0433. The DFT calculations of compounds (5) and (6) have been carried out to ascertain the stability of Z-conformer. The in vitro antimicrobial activity of all the compounds (1-6) was evaluated by the disk diffusion method against gram +ve and gram -ve microorganism and fungal strains. The MIC of the synthesized compounds was determined by agar well diffusion method in 96-well microtiter plate. All the synthesized compounds were also screened for their free radical scavenging activity by DPPH method.

  13. Radiokinetic study on nucleation process of 65Zn(OH)2, 65Zn3(PO4)2 and 51CrPO4 crystals in gelatin and agar

    NASA Astrophysics Data System (ADS)

    Cecal, Al; Palamaru, M.; Chisca, S.; Balan, A.

    1999-01-01

    The nucleation process of 65Zn(OH)2, 65Zn3(PO4)2, and 51CrPO4 crystals in gelatin and agar was studied by using radioactive tracers. The diffusion rate, constants for 65Zn2+ and 51Cr3+ cations through gel, and the reaction rate constants of nucleation process as well as the beginning time of crystal appearance were established. It was found that the reaction rate constant of the low-soluble crystal is higher, and consequently, in a given colloidal medium this parameter varies as follows: k * Zn(PO4)2> k * Zn(OH) 2> k * CrPO 4

  14. Radiokinetic study on nucleation process of 65Zn(OH)2, 65Zn3(PO4)2 and 51CrPO4 crystals in gelatin and agar

    NASA Astrophysics Data System (ADS)

    Cecal, Al; Palamaru, M.; Chisca, S.; Balan, A.

    1999-01-01

    The nucleation process of 65Zn(OH)2, 65Zn3(PO4)2, and 51CrPO4 crystals in gelatin and agar was studied by using radioactive tracers. The diffusion rate, constants for 65Zn2+ and 51Cr3+ cations through gel, and the reaction rate constants of nucleation process as well as the beginning time of crystal appearance were established. It was found that the reaction rate constant of the low-soluble crystal is higher, and consequently, in a given colloidal medium this parameter varies as follows: k * Zn(PO4)2>k * Zn(OH) 2>k * CrPO 4

  15. A novel bioassay using root re-growth in Lemna.

    PubMed

    Park, Areum; Kim, Youn-Jung; Choi, Eun-Mi; Brown, Murray T; Han, Taejun

    2013-09-15

    A new phytotoxicity test method based on root elongation of three Lemna species (Lemna gibba, L. minor, and L. paucicostata) has been developed. Tests with aquatic plants have, typically, favored measurements on fronds (e.g. frond number, area, biomass) rather than on roots, due, in part, to issues associated with handling fragile roots and the time-consuming procedures of selecting roots with identical root lengths. The present method differs in that roots were excised prior to exposure with subsequent measurements on newly developed roots. Results show that there were species-specific difference in sensitivity to the five metals tested (Ag, Cd, Cr, Cu and Hg), with Ag being the most toxic (EC50=5.3-37.6 μgL(-1)) to all three species, and Cr the least toxic for L. gibba and L. minor (1148.3 and 341.8 μgL(-1), respectively) and Cu for L. paucicostata (470.4 μgL(-1)). Direct comparisons were made with measurements of frond area, which were found to be less sensitive. More generally, root re-growth was shown to reflect the toxic responses of all three Lemna species to these five important metals. The root growth bioassay differs from three internationally standardized methods (ISO, OCED and US EPA) in that it is completed in 48 h, the required volume of test solutions is only 3 ml and non-axenic plants are used. Our results show that the Lemna root method is a simple, rapid, cost-effective, sensitive and precise bioassay to assess the toxic risks of metals and has practical application for monitoring municipal and industrial waste waters where metals are common constituents. PMID:23917640

  16. Comprehensive integration of homogeneous bioassays via centrifugo-pneumatic cascading.

    PubMed

    Godino, Neus; Gorkin, Robert; Linares, Ana V; Burger, Robert; Ducrée, Jens

    2013-02-21

    This work for the first time presents the full integration and automation concept for a range of bioassays leveraged by cascading a centrifugo-pneumatic valving scheme to sequentially move several liquids through shared channel segments for multi-step sample preparation into the detection zone. This novel centrifugo-pneumatic liquid handling significantly simplifies system manufacture by obviating the need for complex surface functionalization procedures or hybrid material integration, as it is common in conventional valving methods such as capillary burst valves or sacrificial valves. Based on the centrifugo-pneumatic valving scheme, this work presents a toolkit of operational elements implementing liquid loading/transfer, metering, mixing and sedimentation in a microstructured polymer disc. As a proof of concept for the broad class of homogeneous bioassays, the full integration and automation of a colorimetric nitrate/nitrite test for the detection of clinically relevant nitric oxide (NO) in whole blood is implemented. First, 40 μL of plasma is extracted from a 100 μL sample of human blood, incubated for one hour with the enzymatic mixture (60 μL), and finally reacted with 100 μL of colorimetric (Greiss) reagents. Following just a single loading phase at the beginning of the process, all of these steps are automated through the centrifugo-pneumatic cascade with a high level of flow control and synchronization. Our system shows good correlation with controls up to 50 μM of nitrate, which adequately covers the healthy human range (4 to 45.3 μM). PMID:23250328

  17. Evolving BioAssay Ontology (BAO): modularization, integration and applications

    PubMed Central

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  18. Evolving BioAssay Ontology (BAO): modularization, integration and applications.

    PubMed

    Abeyruwan, Saminda; Vempati, Uma D; Küçük-McGinty, Hande; Visser, Ubbo; Koleti, Amar; Mir, Ahsan; Sakurai, Kunie; Chung, Caty; Bittker, Joshua A; Clemons, Paul A; Brudz, Steve; Siripala, Anosha; Morales, Arturo J; Romacker, Martin; Twomey, David; Bureeva, Svetlana; Lemmon, Vance; Schürer, Stephan C

    2014-01-01

    The lack of established standards to describe and annotate biological assays and screening outcomes in the domain of drug and chemical probe discovery is a severe limitation to utilize public and proprietary drug screening data to their maximum potential. We have created the BioAssay Ontology (BAO) project (http://bioassayontology.org) to develop common reference metadata terms and definitions required for describing relevant information of low-and high-throughput drug and probe screening assays and results. The main objectives of BAO are to enable effective integration, aggregation, retrieval, and analyses of drug screening data. Since we first released BAO on the BioPortal in 2010 we have considerably expanded and enhanced BAO and we have applied the ontology in several internal and external collaborative projects, for example the BioAssay Research Database (BARD). We describe the evolution of BAO with a design that enables modeling complex assays including profile and panel assays such as those in the Library of Integrated Network-based Cellular Signatures (LINCS). One of the critical questions in evolving BAO is the following: how can we provide a way to efficiently reuse and share among various research projects specific parts of our ontologies without violating the integrity of the ontology and without creating redundancies. This paper provides a comprehensive answer to this question with a description of a methodology for ontology modularization using a layered architecture. Our modularization approach defines several distinct BAO components and separates internal from external modules and domain-level from structural components. This approach facilitates the generation/extraction of derived ontologies (or perspectives) that can suit particular use cases or software applications. We describe the evolution of BAO related to its formal structures, engineering approaches, and content to enable modeling of complex assays and integration with other ontologies and

  19. Using lone star ticks, Amblyomma americanum (Acari: Ixodidae) in in vitro laboratory bioassays of repellents: dimensions, duration, and variability

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The in vitro bioassay is an important tool in repellent discovery and development, with a variety of bioassays used in recent years. Several factors, such as the dimensions and configuration of test surfaces and duration of tick exposure, can influence the outcome of bioassays. We tested two tick re...

  20. Mechanical and water barrier properties of agar/κ-carrageenan/konjac glucomannan ternary blend biohydrogel films.

    PubMed

    Rhim, Jong-Whan; Wang, Long-Feng

    2013-07-01

    Multicomponent hydrogel films composed of agar, κ-carrageenan, konjac glucomannan powder, and nanoclay (Cloisite(®) 30B) were prepared and their mechanical and water barrier properties such as water vapor permeability (WVP), water contact angle (CA), water solubility (WS), water uptake ratio (WUR), water vapor uptake ratio (WVUR) were determined. Mechanical, water vapor barrier, and water resistance properties of the ternary blend film exhibited middle range of individual component films, however, they increased significantly after formation of nanocomposite with the clay. Especially, the water holding capacity of the ternary blend biopolymer films increased tremendously, from 800% to 1681% of WUR for agar and κ-carrageenan films up to 5118% and 5488% of WUR for the ternary blend and ternary blend nanocomposite films, respectively. Water vapor adsorption behavior of films was also tested by water vapor adsorption kinetics and water vapor adsorption isotherms test. Preliminary test result for fresh spinach packaging revealed that the ternary blend biohydrogel films had a high potential for the use as an antifogging film for packaging highly respiring agricultural produce. In addition, the ternary blend nanocomposite film showed an antimicrobial activity against Gram-positive bacteria, Listeria monocytogenes. PMID:23688456

  1. 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

  2. Evaluation of a Modified Cefsulodin-Irgasan-Novobiocin Agar for Isolation of Yersinia spp

    PubMed Central

    Tan, Lai Kuan; Ooi, Peck Toung; Carniel, Elisabeth; Thong, Kwai Lin

    2014-01-01

    Y. enterocolitica and Y. pseudotuberculosis are important food borne pathogens. However, the presence of competitive microbiota makes the isolation of Y. enterocolitica and Y. pseudotuberculosis from naturally contaminated foods difficult. We attempted to evaluate the performance of a modified Cefsulodin-Irgasan-Novobiocin (CIN) agar in the differentiation of Y. enterocolitica from non-Yersinia species, particularly the natural intestinal microbiota. The modified CIN enabled the growth of Y. enterocolitica colonies with the same efficiency as CIN and Luria-Bertani agar. The detection limits of the modified CIN for Y. enterocolitica in culture medium (10 cfu/ml) and in artificially contaminated pork (104 cfu/ml) were also comparable to those of CIN. However, the modified CIN provided a better discrimination of Yersinia colonies from other bacteria exhibiting Yersinia-like colonies on CIN (H2S-producing Citrobacter freundii, C. braakii, Enterobacter cloacae, Aeromonas hydrophila, Providencia rettgeri, and Morganella morganii). The modified CIN exhibited a higher recovery rate of Y. enterocolitica from artificially prepared bacterial cultures and naturally contaminated samples compared with CIN. Our results thus demonstrated that the use of modified CIN may be a valuable means to increase the recovery rate of food borne Yersinia from natural samples, which are usually contaminated by multiple types of bacteria. PMID:25170941

  3. Relative value of selective group A streptococcal agar incubated under different atmospheres.

    PubMed Central

    Pacifico, L; Ranucci, A; Ravagnan, G; Chiesa, C

    1995-01-01

    A commercially available selective group A streptococcal agar (ssA) was evaluated for the recovery of group A streptococci (GAS) in comparison with recovery from simultaneous cultures on conventional sheep blood agar (SBA). Both sets of plates were incubated in air, 5% CO2, and anaerobically for 48 h, with a first reading taken at 24 h. A total of 402 (67.0%) GAS were isolated from the 600 specimens that were submitted. Recovery of GAS was significantly greater after 48 h of incubation than after 24 h of incubation for each medium-atmosphere combination. After 48 h of incubation, the sensitivities of GAS detection obtained by each culture technique were as follows: ssA-anaerobic atmosphere, 98.5%; SBA-anaerobic atmosphere, 89.5%; ssA-CO2 atmosphere, 88.0%; SBA-air, 86.5%; SBA-CO2 atmosphere, 82.0%; and ssA-air, 74.6%. There were no cultures positive in air or CO2 which were not positive anaerobically on either medium. The increased sensitivity of detecting positive GAS cultures when incubation was done in an ssA-anaerobic atmosphere for 48 h uncovered patients truly infected with the organisms. PMID:7494053

  4. 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

  5. Evaluation of a modified Cefsulodin-Irgasan-Novobiocin agar for isolation of Yersinia spp.

    PubMed

    Tan, Lai Kuan; Ooi, Peck Toung; Carniel, Elisabeth; Thong, Kwai Lin

    2014-01-01

    Y. enterocolitica and Y. pseudotuberculosis are important food borne pathogens. However, the presence of competitive microbiota makes the isolation of Y. enterocolitica and Y. pseudotuberculosis from naturally contaminated foods difficult. We attempted to evaluate the performance of a modified Cefsulodin-Irgasan-Novobiocin (CIN) agar in the differentiation of Y. enterocolitica from non-Yersinia species, particularly the natural intestinal microbiota. The modified CIN enabled the growth of Y. enterocolitica colonies with the same efficiency as CIN and Luria-Bertani agar. The detection limits of the modified CIN for Y. enterocolitica in culture medium (10 cfu/ml) and in artificially contaminated pork (10(4) cfu/ml) were also comparable to those of CIN. However, the modified CIN provided a better discrimination of Yersinia colonies from other bacteria exhibiting Yersinia-like colonies on CIN (H2S-producing Citrobacter freundii, C. braakii, Enterobacter cloacae, Aeromonas hydrophila, Providencia rettgeri, and Morganella morganii). The modified CIN exhibited a higher recovery rate of Y. enterocolitica from artificially prepared bacterial cultures and naturally contaminated samples compared with CIN. Our results thus demonstrated that the use of modified CIN may be a valuable means to increase the recovery rate of food borne Yersinia from natural samples, which are usually contaminated by multiple types of bacteria. PMID:25170941

  6. Inactivation of Listeria monocytogenes on agar and processed meat surfaces by atmospheric pressure plasma jets.

    PubMed

    Lee, Hyun Jung; Jung, Heesoo; Choe, Wonho; Ham, Jun Sang; Lee, Jun Heon; Jo, Cheorun

    2011-12-01

    An apparatus for generating atmospheric pressure plasma (APP) jet was used to investigate the inactivation of Listeria monocytogenes on the surface of agar plates and slices of cooked chicken breast and ham. He, N₂ (both 7 L/min), and mixtures of each with O₂ (0.07 L/min) were used to produce the plasma jets. After treatment for 2 min with APP jets of He, He + O₂, N₂, or N₂ + O₂, the numbers of L. monocytogenes on agar plates were reduced by 0.87, 4.19, 4.26, and 7.59 log units, respectively. Similar treatments reduced the L. monocytogenes inoculated onto sliced chicken breast and ham by 1.37 to 4.73 and 1.94 to 6.52 log units, respectively, according to the input gas used with the N₂ + O₂ mixture being the most effective. Most APP jets reduced the numbers of aerobic bacteria on the meat surfaces to <10² CFU/g, and the numbers remained below that level of detection after storage at 10 °C for 7 days. The results indicate that APP jets are effective for the inactivation of L. monocytogenes on sliced meats and for prolonging the shelf-life of such foods. PMID:21925030

  7. Abolition of Swarming of Proteus by p-Nitrophenyl Glycerin: Application to Blood Agar Media

    PubMed Central

    Williams, Fred D.

    1973-01-01

    Comparative plate counts were made of Staphylococcus aureus and Streptococcus pyogenes growing on blood agar supplemented with individual chemicals to abolish the swarming of Proteus. B-phenylethanol, sodium azide, and p-nitrophenyl glycerin (PNPG) were used as anti-swarm agents. Each anti-swarm agent effectively abolished swarming for 24 h, but azide failed to control swarming for longer periods of incubation. In addition, azide displayed growth inhibition towards the staphylococci and streptococci resulting in no hemolysis and reduced viable cell numbers with the streptococci. Phenylethanol showed reduced viable cell numbers with the streptococci and unreliable hemolytic reactions. At 0.1 to 0.3 mM, PNPG proved to be a superior anti-swarm agent in that it showed no growth inhibition and allowed normal hemolysis, but abolished swarming for extended periods of time. When laboratory strains of Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa. Listeria monocytogenes, and Vibrio cholerae were screened on a blood agar medium containing 0.1 mm PNPG, they displayed similar growth and hemolytic characteristics to the identical medium without PNPG. PMID:4715553

  8. Colonic cell growth and mucin degradation in rats fed diets containing various levels of beta-carotene with and without dietary agar.

    PubMed

    Hwa, S H; Shiau, S Y

    1993-06-01

    1. To either an agar-containing diet or an agar-free diet, 0, 0.3 and 2.0 mg/100 g of beta-carotene were incorporated and fed to groups of five rats for 28 days. 2. Weight gain and food consumption of rats fed different dietary groups did not show a significant difference (P > 0.05). 3. Colon weight, colonic mucosal DNA and RNA were generally higher in rats fed agar diets than rats fed agar-free diets at either beta-carotene supplementation level. 4. Mucinase activity was higher (P < 0.05) in rats fed the agar diet than in rats fed an agar-free diet without beta-carotene. However, the difference was not observed (P > 0.05) when beta-carotene was incorporated. 5. These data suggest that colonic mucin degradation in rats fed an agar diet decreased when the dietary beta-carotene inclusion level increased. PMID:7687211

  9. Vaneless diffusers

    NASA Astrophysics Data System (ADS)

    Senoo, Y.

    The influence of vaneless diffusers on flow in centrifugal compressors, particularly on surge, is discussed. A vaneless diffuser can demonstrate stable operation in a wide flow range only if it is installed with a backward leaning blade impeller. The circumferential distortion of flow in the impeller disappears quickly in the vaneless diffuser. The axial distortion of flow at the diffuser inlet does not decay easily. In large specific speed compressors, flow out of the impeller is distorted axially. Pressure recovery of diffusers at distorted inlet flow is considerably improved by half guide vanes. The best height of the vanes is a little 1/2 diffuser width. In small specific speed compressors, flow out of the impeller is not much distorted and pressure recovery can be predicted with one-dimensional flow analysis. Wall friction loss is significant in narrow diffusers. The large pressure drop at a small flow rate can cause the positive gradient of the pressure-flow rate characteristic curve, which may cause surging.

  10. An in vitro rainbow trout cell bioassay for AhR-mediated toxins

    SciTech Connect

    Richter, C.A.; Giesy, J.P.; Denison, M.S.

    1995-12-31

    The toxicity of PCBs, dioxins, and other halogenated aromatic hydrocarbons (HAHS) at environmentally relevant concentrations is in large part mediated through the aromatic hydrocarbon receptor (AhR). Bioassays which measure the activity of genes regulated by the receptor provide an integrative measure of the total AhR-mediated toxicity of a sample. The authors have recently developed and characterized a bioassay using recombinant rainbow trout hepatoma cells containing the firefly luciferase reporter gene under the regulation of the AhR. The cell line is designated Remodulated Lightning Trout (RLT). The RLT bioassay is relevant to fish, and is useful as a rapid screening device, a guide for chemical analysis, and a tool for studies of the AhR mechanism. The responses of the RLT cell line to various PCB congeners are similar to responses of in vivo fish bioassays. The authors now report on the responses of the bioassay to dioxins, dibenzofurans, and other related compounds as compared to in vivo fish bioassays. The authors will also report on the utility of the RLT bioassay in measuring the total TEQ of complex mixtures.

  11. Sensitive, Rapid, and Specific Bioassay for the Determination of Antilipogenic Compounds

    PubMed Central

    Ulitzur, S.; Goldberg, I.

    1977-01-01

    A sensitive and rapid bioassay for the determination of the antilipogenic compounds cerulenin and CM-55 is described. The bioassay is based on the inhibitory effect of cerulenin and CM-55 on the in vivo luminescence of an aldehyde-requiring mutant of the marine bacterium Beneckea harveyi. A total quantity as low as 0.1 μg of cerulenin can be determined within 15 min with an error of ±2%. The bioassay, as presented, is specific for compounds that are known to inhibit fatty acid biosynthesis and, as such, it might be used as a general screening method for the detection of antilipogenic compounds. PMID:303076

  12. Diffusion barriers

    NASA Technical Reports Server (NTRS)

    Nicolet, M. A.

    1983-01-01

    The choice of the metallic film for the contact to a semiconductor device is discussed. One way to try to stabilize a contact is by interposing a thin film of a material that has low diffusivity for the atoms in question. This thin film application is known as a diffusion barrier. Three types of barriers can be distinguished. The stuffed barrier derives its low atomic diffusivity to impurities that concentrate along the extended defects of a polycrystalline layer. Sacrificial barriers exploit the fact that some (elemental) thin films react in a laterally uniform and reproducible fashion. Sacrificial barriers have the advantage that the point of their failure is predictable. Passive barriers are those most closely approximating an ideal barrier. The most-studied case is that of sputtered TiN films. Stuffed barriers may be viewed as passive barriers whose low diffusivity material extends along the defects of the polycrystalline host.

  13. Diffuse radiation

    NASA Technical Reports Server (NTRS)

    1981-01-01

    A diffuse celestial radiation which is isotropic at least on a course scale were measured from the soft X-ray region to about 150 MeV, at which energy the intensity falls below that of the galactic emission for most galactic latitudes. The spectral shape, the intensity, and the established degree of isotropy of this diffuse radiation already place severe constraints on the possible explanations for this radiation. Among the extragalactic theories, the more promising explanations of the isotropic diffuse emission appear to be radiation from exceptional galaxies from matter antimatter annihilation at the boundaries of superclusters of galaxies of matter and antimatter in baryon symmetric big bang models. Other possible sources for extragalactic diffuse gamma radiation are discussed and include normal galaxies, clusters of galaxies, primordial cosmic rays interacting with intergalactic matter, primordial black holes, and cosmic ray leakage from galaxies.

  14. First Report of Crown Gall Caused by Agrobacterium sp. on Diffuse Knapweed (Centaurea diffusa)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A specimen of diffuse knapweed (Centaurea diffusa, DK) with crown gall-like symptoms was collected July 27, 2004, in Mosier, Wasco Co., OR (N 45.6842, W 121.4021), and sent to the USDA at Ft. Detrick, MD, for identification. A bacterium was isolated on Potato Dextrose Agar that caused hyperplasia a...

  15. Isolation and characterization of agar-digesting Vibrio species from the rotten thallus of Gracilariopsis heteroclada Zhang et Xia.

    PubMed

    Martinez, Joval N; Padilla, Philip Ian P

    2016-08-01

    Gracilariopsis heteroclada Zhang et Xia (Gracilariaceae, Rhodophyta) is one of the most studied marine seaweeds due to its economic importance. This has been cultivated extensively on commercial scale in the Philippines and other Asian countries. However, sustainable production of G. heteroclada in the Philippines could not be maximized due to the occurrence of rotten thallus disease. Thus, isolation and characterization of agar-digesting bacteria from the rotten thalli of G. heteroclada was conducted. A total of seven representative bacterial isolates were randomly selected based on their ability to digest agar as evidenced by the formation of depressions around the bacterial colonies on nutrient agar plates supplemented with 1.5% NaCl and liquefaction of agar. Gram-staining and biochemical characterization revealed that isolates tested were gram-negative rods and taxonomically identified as Vibrio parahaemolyticus (86-99.5%) and Vibrio alginolyticus (94.2-97.7%), respectively. It is yet to be confirmed whether these agar-digesting vibrios are involved in the induction and development of rotten thallus disease in G. heteroclada in concomitance with other opportunistic bacterial pathogens coupled with adverse environmental conditions. PMID:27285614

  16. Enumeration of food-borne Clostridium perfringens in egg yolk-free tryptose-sulfite-cycloserine agar.

    PubMed

    Hauschild, A H; Hilsheimer, R

    1974-03-01

    The SFP (Shahidi-Ferguson perfringens), TSC (tryptose-sulfite-cycloserine), EY (egg yolk)-free TSC, and OPSP (oleandomycin-polymyxin-sulfadiazine perfringens) agars have been tested for their suitability to enumerate Clostridium perfringens in naturally contaminated foods. Complete recoveries of C. perfringens were obtained in each of the four media, but only the TSC and EY-free TSC agars were sufficiently selective to ensure subsequent confirmatory tests without interference from facultative anaerobes. Because of some disadvantages associated with the use of egg yolk, EY-free TSC agar is recommended for enumeration of C. perfringens in foods. Several conditions for convenient shipment of foods and C. perfringens isolates with minimum loss of viability have been tested. The highest viable counts were preserved when foods were mixed 1:1 (wt/vol) with 20% glycerol and kept in a container with dry ice. Isolated C. perfringens strains remained viable for at least 2 weeks at ambient temperatures on blood agar slopes with a 2% agar overlay in screw-cap culture tubes. PMID:4363368

  17. Enumeration of Food-Borne Clostridium perfringens in Egg Yolk-Free Tryptose-Sulfite-Cycloserine Agar

    PubMed Central

    Hauschild, A. H. W.; Hilsheimer, R.

    1974-01-01

    The SFP (Shahidi-Ferguson perfringens), TSC (tryptose-sulfite-cycloserine), EY (egg yolk)-free TSC, and OPSP (oleandomycin-polymyxin-sulfadiazine perfringens) agars have been tested for their suitability to enumerate Clostridium perfringens in naturally contaminated foods. Complete recoveries of C. perfringens were obtained in each of the four media, but only the TSC and EY-free TSC agars were sufficiently selective to ensure subsequent confirmatory tests without interference from facultative anaerobes. Because of some disadvantages associated with the use of egg yolk, EY-free TSC agar is recommended for enumeration of C. perfringens in foods. Several conditions for convenient shipment of foods and C. perfringens isolates with minimum loss of viability have been tested. The highest viable counts were preserved when foods were mixed 1:1 (wt/vol) with 20% glycerol and kept in a container with dry ice. Isolated C. perfringens strains remained viable for at least 2 weeks at ambient temperatures on blood agar slopes with a 2% agar overlay in screw-cap culture tubes. PMID:4363368

  18. The effects of nutrient chemotaxis on bacterial aggregation patterns with non-linear degenerate cross diffusion

    NASA Astrophysics Data System (ADS)

    Leyva, J. Francisco; Málaga, Carlos; Plaza, Ramón G.

    2013-11-01

    This paper studies a reaction-diffusion-chemotaxis model for bacterial aggregation patterns on the surface of thin agar plates. It is based on the non-linear degenerate cross diffusion model proposed by Kawasaki et al. (1997) [5] and it includes a suitable nutrient chemotactic term compatible with such type of diffusion, as suggested by Ben-Jacob et al. (2000) [20]. An asymptotic estimation predicts the growth velocity of the colony envelope as a function of both the nutrient concentration and the chemotactic sensitivity. It is shown that the growth velocity is an increasing function of the chemotactic sensitivity. High resolution numerical simulations using Graphic Processing Units (GPUs), which include noise in the diffusion coefficient for the bacteria, are presented. The numerical results verify that the chemotactic term enhances the velocity of propagation of the colony envelope. In addition, the chemotaxis seems to stabilize the formation of branches in the soft-agar, low-nutrient regime.

  19. Characteristics of rat megakaryocyte colonies and their progenitors in agar culture

    SciTech Connect

    Kellar, K.L.; Rolovic, Z.; Evatt, B.L.; Sewell, E.T.

    1985-11-01

    The characteristics of megakaryocyte colonies that develop from megakaryocyte progenitors of rat bone marrow stimulated by rat spleen-conditioned medium (SCM) in agar culture were investigated. Colony frequency was optimal on day 7 and increased relative to both the number of cells plated and the concentration of SCM used. Colonies were categorized as small cell and big cell. Small-cell colonies had a greater proliferative potential, with a mean of 25 cells/colony. Big-cell colonies averaged 15 cells/colony. The ratio of big-cell to small-cell colonies was 0.69 +/- 0.29. Granulocyte-macrophage colonies, which were also stimulated by SCM, accounted for 70% +/- 15% of the total colonies in the cultures. Cytocidal experiments with tritiated thymidine reduced megakaryocyte colony formation by 45% and granulocyte-macrophage colony formation by 21%. The properties of rat, mouse, and human megakaryocyte progenitors as assayed in vitro are compared.

  20. Comparison of four commercial brucella agar media for growth of anaerobic organisms.

    PubMed Central

    Mangels, J I; Douglas, B P

    1989-01-01

    Four different commercial brucella blood agar plating media (Anaerobe Systems, BBL Microbiology Systems, Remel, and Scott Laboratories) were compared for the abilities to recover anaerobic organisms from clinical specimens and to support the growth of American Type Culture Collection anaerobic stock cultures. Following 24 h of incubation in an anaerobe chamber, Anaerobe Systems prereduced, anaerobically sterilized brucella plates yielded 63% of the total clinical anaerobe isolates, the Scott medium yielded 51%, the Remel medium yielded 42%, and the BBL medium yielded 37%. Poor growth of Peptostreptococcus magnus, P. anaerobius, Fusobacterium necrophorum, F. nucleatum, and pigmented Bacteroides spp. was observed on brucella media obtained from BBL, Remel, and Scott. Data obtained with stock anaerobic cultures showed that Anaerobe Systems plates yielded good growth and produced a larger colony size with all of the strains tested in 1 day, whereas poor growth of Peptostreptococcus spp., B. melaninogenicus, and Fusobacterium spp. was noted on brucella media from BBL, Remel, and Scott. PMID:2584378

  1. Nocardia pigrifrangens sp. nov., a novel actinomycete isolated from a contaminated agar plate.

    PubMed

    Wang, Liming; Zhang, Yamei; Huang, Ying; Maldonado, Luis A; Liu, Zhiheng; Goodfellow, Michael

    2004-09-01

    A polyphasic study was undertaken to establish the taxonomic position of an actinomycete strain isolated from a contaminated agar plate. The strain, designated 7031T, had morphological and chemotaxonomic properties typical of the genus Nocardia. An almost-complete 16S rRNA gene sequence determined for the strain was aligned with available sequences for nocardiae, and phylogenetic trees were inferred using three tree-generating algorithms. Strain 7031T clustered with the type strains of Nocardia carnea and Nocardia flavorosea, showing low 16S rRNA gene sequence similarities to these species (97.2 and 97.5 %, respectively). The strain was also distinguished from the closest species by a range of phenotypic properties. It is proposed that the strain be recognized as a novel species of Nocardia, Nocardia pigrifrangens sp. nov., the type strain of which is 7031T (= AS 4.1808T = JCM 11884T). PMID:15388728

  2. Injection of Acanthaster planci with thiosulfate-citrate-bile-sucrose agar (TCBS). I. Disease induction.

    PubMed

    Rivera-Posada, J A; Pratchett, M; Cano-Gómez, A; Arango-Gómez, J D; Owens, L

    2011-12-01

    This is the first report of the successful induction of a transmissible disease in the coral-eating crown-of-thorns starfish Acanthaster planci (COTS). Injection of thiosulfate-citrate-bile-sucrose agar (TCBS) culture medium into COTS induced a disease characterized by discoloured and necrotic skin, ulcerations, loss of body turgor, accumulation of colourless mucus on many spines especially at their tip, and loss of spines. Blisters on the dorsal integument broke through the skin surface and resulted in large, open sores that exposed the internal organs. Oedema and reddened digestive tissues and destruction of connective fibers were common. Moreover, healthy COTS in contact with these infected animals also displayed signs of disease and died within 24 h. TCBS induced 100% mortality in injected starfish. There was no introduction of new pathogens into the marine environment. TCBS promoted the growth of COTS' naturally occurring Vibrionales to high densities with subsequent symbiont imbalance followed by disease and death. PMID:22303625

  3. [Cryofractographic study of intercellular junctions in the populations of agar-cultivated Bordetella pertussis].

    PubMed

    Vysotskiĭ, V V; Vaisman, I Sh; Efimova, O G; Chemurzieva, N V

    1985-09-01

    The characteristic feature of replicas obtained from the freeze-fractures of B. pertussis unfixed cultures developing on casein charcoal agar for 1-7 days is the associative growth of highly polymorphic cells, ensured by the ramified system of intercellular connections (IC) formed by the derivatives of the outer layers of the cell wall. This proves that the associative location of bacterial cells, linked by numerous IC, in the preparation is not the artefact appearing in the process of their chemical fixation. In replicas obtained from the freeze-fractures of B. pertussis cultures, previously fixed with glutaraldehyde, osmic acid and uranyl acetate, oval cells with the cytoplasm having a relatively homogeneous structure and with the smoothed-out three-layer cell wall prevail. As a rule, IC are limited to the sites of direct contacts between individual cells. PMID:2866645

  4. Agar gel immunodiffusion assay to detect antibodies to Type A influenza virus.

    PubMed

    Jenson, Terra A

    2014-01-01

    The agar gel immunodiffusion (AGID) test is used to detect antibodies to Type A influenza group-specific antigens, i.e., the ribonucleoprotein (RNP) and matrix (M) proteins. Therefore, this test will detect antibodies to all influenza A virus subtypes. AGID is commonly used to screen poultry flocks for avian influenza virus infection. The AGID is a simple and economical serological test. All serological testing has its advantages and disadvantages which should be considered before choosing the optimal test for the laboratory needs. Each laboratory must evaluate the laboratory's resources, the volume of testing, the goal of testing, how the test results are used and what types of samples are being tested in order to select the optimal test. PMID:24899427

  5. Growth of Bacillus cereus on solid media as affected by agar, sodium chloride, and potassium sorbate.

    PubMed

    Stecchini, M L; Del Torre, M; Donda, S; Maltini, E

    2000-07-01

    The effect of two independent variables: microstructure, as modified by the agar content (1.0, 4.0, 7.0%), and water activity (a(w)), as modified by the NaCl content (0.5, 2.5, 4.5%), in the absence or in the presence of potassium sorbate (0.0; 2,000 ppm) on Bacillus cereus growth on solid media was studied. The time to visible growth (TVG) and the radial growth rate (RGR) of colonies were evaluated. TVG was not affected by microstructure and K-sorbate, although when a(w) was reduced, TVG tended to increase. RGR depended on linear effects of microstructure and a(w) variables and their interaction. When K-sorbate was added to cultural media, RGR was reduced significantly. However, in the presence of K-sorbate, RGR was found to change only when a(w) vas varied. PMID:10914662

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

    PubMed

    Daly, J G; Stevenson, R M

    1993-07-01

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

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

    PubMed Central

    Daly, J. G.; Stevenson, R. M. W.

    1993-01-01

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

  8. Charcoal agar, a new growth medium for the fish disease bacterium Renibacterium salmoninarum.

    PubMed Central

    Daly, J G; Stevenson, R M

    1985-01-01

    Charcoal is an effective replacement for serum in media for the isolation and culture of Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmonid fish. The medium, KDM-C, contains 10 g of peptone, 0.5 g of yeast extract, 1 g of L-cysteine hydrochloride, 1 g of activated charcoal, and 15 g of agar per liter and is adjusted to pH 6.8 with NaOH before autoclaving. Eight strains of R. salmoninarum grew from dilute inocula as well on KDM-C as on a standard serum-containing medium (KDM-2). The medium was effective for both primary isolations from fish and repeated transfers and has potential value for antigen preparation and physiological studies. Images PMID:4083882

  9. 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. PMID:25944532

  10. Evolutionary consequences of putative intra-and interspecific hybridization in agaric fungi.

    PubMed

    Hughes, Karen W; Petersen, Ronald H; Lodge, D Jean; Bergemann, Sarah E; Baumgartner, Kendra; Tulloss, Rodham E; Lickey, Edgar; Cifuentes, Joaquin

    2013-01-01

    Agaric fungi of the southern Appalachian Mountains including Great Smoky Mountains National Park are often heterozygous for the rDNA internal transcribed spacer region (ITS) with >42% of collections showing some heterozygosity for indels and/or base-pair substitutions. For these collections, intra-individual haplotype divergence is typically less than 2%, but for 3% of these collections intra-individual haplotype divergence exceeds that figure. We hypothesize that high intra-individual haplotype divergence is due to hybridization between agaric fungi with divergent haplotypes, possibly migrants from geographically isolated glacial refugia. Four species with relatively high haplotype divergence were examined: Armillaria mellea, Amanita citrina f. lavendula, Gymnopus dichrous and the Hygrocybe flavescens/chlorophana complex. The ITS region was sequenced, haplotypes of heterozygotes were resolved through cloning, and phylogenetic analyses were used to determine the outcome of hybridization events. Within Armillaria mellea and Amanita citrina f. lavendula, we found evidence of interbreeding and recombination. Within G. dichrous and H. flavescens/chlorophana, hybrids were identified but there was no evidence for F2 or higher progeny in natural populations suggesting that the hybrid fruitbodies might be an evolutionary dead end and that the genetically divergent Mendelian populations from which they were derived are, in fact, different species. The association between ITS haplotype divergence of less than 5% (Armillaria mellea = 2.6% excluding gaps; Amanita citrina f. lavendula = 3.3%) with the presence of putative recombinants and greater than 5% (Gymnopus dichrous = 5.7%; Hygrocybe flavescens/chlorophana = 14.1%) with apparent failure of F1 hybrids to produce F2 or higher progeny in populations may suggest a correlation between genetic distance and reproductive isolation. PMID:23928423

  11. Analyzing bioassay data using Bayesian methods -- A primer

    SciTech Connect

    Miller, G.; Inkret, W.C.; Schillaci, M.E.

    1997-10-16

    The classical statistics approach used in health physics for the interpretation of measurements is deficient in that it does not allow for the consideration of needle in a haystack effects, where events that are rare in a population are being detected. In fact, this is often the case in health physics measurements, and the false positive fraction is often very large using the prescriptions of classical statistics. Bayesian statistics provides an objective methodology to ensure acceptably small false positive fractions. The authors present the basic methodology and a heuristic discussion. Examples are given using numerically generated and real bioassay data (Tritium). Various analytical models are used to fit the prior probability distribution, in order to test the sensitivity to choice of model. Parametric studies show that the normalized Bayesian decision level k{sub {alpha}}-L{sub c}/{sigma}{sub 0}, where {sigma}{sub 0} is the measurement uncertainty for zero true amount, is usually in the range from 3 to 5 depending on the true positive rate. Four times {sigma}{sub 0} rather than approximately two times {sigma}{sub 0}, as in classical statistics, would often seem a better choice for the decision level.

  12. Target organs in chronic bioassays of 533 chemical carcinogens

    SciTech Connect

    Gold, L.S.; Slone, T.H.; Manley, N.B. ); Bernstein, L. )

    1991-06-01

    A compendium of carcinogenesis bioassay results organized by target organ is presented for 533 chemicals that are carcinogenic in at least one species. This compendium is based primarily on experiments in rats or mice; results in hamsters, nonhuman primates, and dogs are also reported. The compendium can be used to identify chemicals that induce tumors at particular sites, and to determine whether target sites are the same for chemicals positive in more than one species. The Carcinogenic Potency Database (CPDB), which includes results of 3969 experiments, is used in the analysis. The published CPDB includes details on each test, and literature references. Chemical carcinogens are reported for 35 different target organs in rats or mice. More than 80% of the carcinogens in each of these species are positive in at least one of the 8 most frequent target sites; liver, lung, mammary gland, stomach, vascular system, kidney, hematopoietic system, and urinary bladder. An analysis is presented of how well one can predict the carcinogenic response in mice from results in rats, or vice versa. Among chemicals tested in both species, 76% of rat carcinogens are positive in mice, and 71% of mouse carcinogens are positive in rats. Prediction is less accurate to the same target site: 52% of rat carcinogens are positive in the same site in mice, and 48% of mouse carcinogens are positive in the same site in rats. The liver is the most frequent site in common between rats and mice.

  13. Using enzyme bioassays as a rapid screen for metal toxicity

    USGS Publications Warehouse

    Choate, LaDonna M.; Ross, P.E.; Blumenstein, E. P.; Ranville, James F.

    2005-01-01

    Mine tailings piles and abandoned mine soils are often contaminated by a suite of toxic metals, which were released in the mining process. Traditionally, toxicity of such areas has been determined by numerous chemical methods including the Toxicity Characteristic Leachate Procedure (TCLP) and traditional toxicity tests using organisms such as the cladoceran Ceriodaphnia dubia. Such tests can be expensive and time-consuming. Enzymatic bioassays may provide an easier, less costly, and more time-effective toxicity screening procedure for mine tailings and abandoned mine soil leachates. This study evaluated the commercially available MetPLATE™ enzymatic toxicity assay test kit. The MetPLATE™ assay uses a modified strain of Escherichia coli bacteria as the test organism. Toxicity is defined by the activity of β-galactosidase enzyme which is monitored colorometrically with a 96-well spectrophotometer. The study used water samples collected from North Fork Clear Creek, a mining influenced water (MIW) located in Colorado. A great benefit to using the MetPLATE™ assay over the TCLP is that it shows actual toxicity of a sample by taking into account the bioavailability of the toxicants rather than simply measuring the metal concentration present. Benefits of the MetPLATE™ assay over the use of C. dubia include greatly reduced time for the testing process (∼2 hours), a more continuous variable due to a greater number of organisms present in each sample (100,000+), and the elimination of need to maintain a culture of organisms at all times.

  14. Bioassays on Illinois waterway dredged material. Final report

    SciTech Connect

    Moore, D.W.; Gibson, A.B.; Dillon, T.M.

    1992-12-01

    Sediment from the Illinois Waterway navigation channel is hydraulically dredged by the US Army Engineer District, Rock Island, and placed in the nearshore environment via pipeline. Water returning to the river can have a high-suspended solids load approaching fluid mud consistency. There is a concern that this return water may exceed the State of Illinois water quality standards for ammonia and have adverse effects on aquatic life. To address these concerns, composite sediment samples and site water collected from selected sites in the Illinois Waterway were evaluated in toxicity tests. Acute (48-hr) toxicity tests were conducted with two species, Pimephales promelas (the fathead minnow) and Daphnia magna (a freshwater cladoceran). A chronic (21-day) toxicity test was also conducted using Daphnia magna. Animals were exposed separately to different concentrations of filtered and unfiltered elutriates prepared from Acute, Cadmium, Daphnia magna, Pimephales promela, Ammonia, Chronic, Elutriate, Sediment, Bioassay, Cladoceran, Fathead minnow. Illinois Waterway edged material. Total ammonia concentrations were measured in all tests and the un-ionized fraction was calculated by adjusting for temperature and pH. Tests were conducted at the US Army Engineer Waterways Experiment Station, Vicksburg, MS. In addition, as part of an interlaboratory effort, a 48-hr acute toxicity test with Pimephales pomelas fry was conducted concurrently by the Hygienic Laboratory of the University of Iowa, Des Moines, IA.

  15. Long-wavelength-emitting nanocrystals for bioassay applications

    NASA Astrophysics Data System (ADS)

    Leppert, Valerie J.; Harvey, Ashley S.; McCool, Geoff D.; Quinlan, Forest T.; Feng, Jun; Shan, Guomin; Stroeve, Pieter; Risbud, Subhash H.; Hammock, Bruce D.; Kennedy, Ian M.

    2002-11-01

    New fluorophores that can be excited using visible or near-infrared radiation are of considerable interest for application in environmental and complex bioassays, where background fluorescence is exacerbated by ultra-violet or blue excitation. Useful labels for biomolecules include infrared emitting semiconductor nanoparticles that can be blue-shifted into the near-infrared and visible through quantum confinement effects, oxides of iron, and rare earth oxides. In this work, the synthesis of 6 nm average diameter lead selenide nanocrystals (well below the Bohr exciton diameter of 92 nm) through a reverse micelle technique; and the synthesis of iron and europium oxides with particles less than 5 nm in diameter by pulsed laser ablation is reported. The europium oxide nanoparticles' emission showed a large Stokes shift (144 nm or 216 nm, depending on excitation wavelength); a narrow, symmetric emission line at 610 nm (FWHM of 8 nm); and long lifetime (300 μs). The Eu2O3 nanoparticles, which were coated with silica for functionalization, displayed a greatly enhanced sensitivity over a conventional ELISA (0.025 ng ml-1 vs. 0.1 ng ml-1) when run in an atrazine immunoassay.

  16. Detection of Organic Compounds with Whole-Cell Bioluminescent Bioassays

    PubMed Central

    Xu, Tingting; Close, Dan; Smartt, Abby; Ripp, Steven

    2015-01-01

    Natural and manmade organic chemicals are widely deposited across a diverse range of ecosystems including air, surface water, groundwater, wastewater, soil, sediment, and marine environments. Some organic compounds, despite their industrial values, are toxic to living organisms and pose significant health risks to humans and wildlife. Detection and monitoring of these organic pollutants in environmental matrices therefore is of great interest and need for remediation and health risk assessment. Although these detections have traditionally been performed using analytical chemical approaches that offer highly sensitive and specific identification of target compounds, these methods require specialized equipment and trained operators, and fail to describe potential bioavailable effects on living organisms. Alternatively, the integration of bioluminescent systems into whole-cell bioreporters presents a new capacity for organic compound detection. These bioreporters are constructed by incorporating reporter genes into catabolic or signaling pathways that are present within living cells and emit a bioluminescent signal that can be detected upon exposure to target chemicals. Although relatively less specific compared to analytical methods, bioluminescent bioassays are more cost-effective, more rapid, can be scaled to higher throughput, and can be designed to report not only the presence but also the bioavailability of target substances. This chapter reviews available bacterial and eukaryotic whole-cell bioreporters for sensing organic pollutants and their applications in a variety of sample matrices. PMID:25084996

  17. Bioassay-based risk assessment of complex mixtures

    SciTech Connect

    Donnelly, K.C.; Safe, S.H.; Randerath, K.; Randerath, E.

    1994-12-31

    To compare the standard chemical-based risk assessment with in vitro genotoxicity assays, two complex environmental mixtures from a wood preserving site were analyzed in the Salmonella/microsome and E. coli prophage induction assays. Using GC/MS, sample 003 was found to contain relatively low levels of polycyclic aromatic hydrocarbons (PNAs) and elevated levels of polychlorinated dibenzo-p-dioxins (PCDDs), while sample 005 had higher levels of PNAs and relatively low levels of PCDDs. The complex mixtures were sequentially extracted with methylene chloride and methanol for analysis in Salmonella, or extracted with 1:1 hexane: acetone mixture for analysis in the prophage induction assay. At a dose of 1.0 mg/plate in Salmonella strain TA98 with metabolic activation, the methanol extract of sample 003 induced 197 net revertants, while sample 005 induced 436 net revertants. In the prophage induction assay, with activation, the hexane:acetone extract of sample 003 induced a fold increase that was slightly lower than that observed with sample 005. The estimated incremental carcinogenic risk for dermal adsorption and ingestion was 1.5E-3 for sample 003, while for sample 005 the estimated risk was 1.5E-2. Thus, the sample which induced the maximum response in both bioassays also had the highest estimated cancer risk. However, the frequency of PNA-DNA adducts in both skin and liver tissues was appreciably higher with sample 005 than with sample 003.

  18. Luminescent Lanthanide Reporters for High-Sensitivity Novel Bioassays.

    SciTech Connect

    Anstey, Mitchell; Fruetel, Julia A.; Foster, Michael E.; Hayden, Carl C.; Buckley, Heather L.; Arnold, John

    2013-09-01

    Biological imaging and assay technologies rely on fluorescent organic dyes as reporters for a number of interesting targets and processes. However, limitations of organic dyes such as small Stokes shifts, spectral overlap of emission signals with native biological fluorescence background, and photobleaching have all inhibited the development of highly sensitive assays. To overcome the limitations of organic dyes for bioassays, we propose to develop lanthanide-based luminescent dyes and demonstrate them for molecular reporting applications. This relatively new family of dyes was selected for their attractive spectral and chemical properties. Luminescence is imparted by the lanthanide atom and allows for relatively simple chemical structures that can be tailored to the application. The photophysical properties offer unique features such as narrow and non-overlapping emission bands, long luminescent lifetimes, and long wavelength emission, which enable significant sensitivity improvements over organic dyes through spectral and temporal gating of the luminescent signal.Growth in this field has been hindered due to the necessary advanced synthetic chemistry techniques and access to experts in biological assay development. Our strategy for the development of a new lanthanide-based fluorescent reporter system is based on chelation of the lanthanide metal center using absorbing chromophores. Our first strategy involves %22Click%22 chemistry to develop 3-fold symmetric chelators and the other involves use of a new class of tetrapyrrole ligands called corroles. This two-pronged approach is geared towards the optimization of chromophores to enhance light output.

  19. Target organs in chronic bioassays of 533 chemical carcinogens.

    PubMed Central

    Gold, L S; Slone, T H; Manley, N B; Bernstein, L

    1991-01-01

    A compendium of carcinogenesis bioassay results organized by target organ is presented for 533 chemicals that are carcinogenic in at least one species. This compendium is based primarily on experiments in rats or mice; results in hamsters, nonhuman primates, and dogs are also reported. The compendium can be used to identify chemicals that induce tumors at particular sites, and to determine whether target sites are the same for chemicals positive in more than one species. The Carcinogenic Potency Database (CPDB), which includes results of 3969 experiments, is used in the analysis. The published CPDB includes details on each test, and literature references. Chemical carcinogens are reported for 35 different target organs in rats or mice. More than 80% of the carcinogens in each of these species are positive in at least one of the 8 most frequent target sites: liver, lung, mammary gland, stomach, vascular system, kidney, hematopoietic system, and urinary bladder. An analysis is presented of how well one can predict the carcinogenic response in mice from results in rats, or vice versa. Among chemicals tested in both species, 76% of rat carcinogens are positive in mice, and 71% of mouse carcinogens are positive in rats. Prediction is less accurate to the same target site: 52% of rat carcinogens are positive in the same site in mice, and 48% of mouse carcinogens are positive in the same site in rats. The liver is the most frequent site in common between rats and mice. PMID:1773795

  20. Bioassay-based risk assessment of complex mixtures

    SciTech Connect

    Donnelly, K.C.; Huebner, H.J.

    1996-12-31

    The baseline risk assessment often plays an integral role in various decision-making processes at Superfund sites. The present study reports on risk characterizations prepared for seven complex mixtures using biological and chemical analysis. Three of the samples (A, B, and C) were complex mixtures of polycyclic aromatic hydrocarbons (PAHs) extracted from coal tar; while four samples extracted from munitions-contaminated soil contained primarily nitroaromatic hydrocarbons. The chemical-based risk assessment ranked sample C as least toxic, while the risk associated with samples A and B was approximately equal. The microbial bioassay was in general agreement for the coal tar samples. The weighted activity of the coal tar extracts in Salmonella was 4,960 for sample C, and 162,000 and 206,000 for samples A and B, respectively. The bacterial mutagenicity of 2,4,6-trinitrotoluene contaminated soils exhibited an indirect correlation with chemical-based risk assessment. The aqueous extract of sample 004 induced 1,292 net revertants in Salmonella, while the estimated risk to ingestion and dermal adsorption was 2E-9. The data indicate that the chemical-based risk assessment accurately predicted the genotoxicity of the PAHs, while the accuracy of the risk assessment for munitions contaminated soils was limited due to the presence of metabolites of TNT degradation. The biological tests used in this research provide a valuable compliment to chemical analysis for characterizing the genotoxic risk of complex mixtures.

  1. Colorimetric paper bioassay for the detection of phenolic compounds.

    PubMed

    Alkasir, Ramiz S J; Ornatska, Maryna; Andreescu, Silvana

    2012-11-20

    A new type of paper based bioassay for the colorimetric detection of phenolic compounds including phenol, bisphenol A, catechol and cresols is reported. The sensor is based on a layer-by-layer (LbL) assembly approach formed by alternatively depositing layers of chitosan and alginate polyelectrolytes onto filter paper and physically entrapping the tyrosinase enzyme in between these layers. The sensor response is quantified as a color change resulting from the specific binding of the enzymatically generated quinone to the multilayers of immobilized chitosan on the paper. The color change can be quantified with the naked eye but a digitalized picture can also be used to provide more sensitive comparison to a calibrated color scheme. The sensor was optimized with respect to the number of layers, pH, enzyme, chitosan and alginate amounts. The colorimetric response was concentration dependent, with a detection limit of 0.86 (±0.1) μg/L for each of the phenolic compounds tested. The response time required for the sensor to reach steady-state color varied between 6 and 17 min depending on the phenolic substrate. The sensor showed excellent storage stability at room temperature for several months (92% residual activity after 260 days storage) and demonstrated good functionality in real environmental samples. A procedure to mass-produce the bioactive sensors by inkjet printing the LbL layers of polyelectrolyte and enzyme on paper is demonstrated. PMID:23113670

  2. USING BIOASSAYS TO EVALUATE THE PERFORMANCE OF EDC RISK MANAGEMENT METHODS

    EPA Science Inventory

    In Superfund risk management research, the performance of risk management techniques is typically evaluated by measuring "the concentrations of the chemicals of concern before and after risk management efforts. However, using bioassays and chemical data provides a more robust und...

  3. COMPARATIVE POTENCY OF COMPLEX MIXTURES: USE OF SHORT-TERM GENETIC BIOASSAYS IN CANCER RISK ASSESSMENT

    EPA Science Inventory

    The primary problem regarding the introduction of new energy sources is whether they will alter the mutagenicity, carcinogenicity and potential human cancer risk from combustion emissions. New risk assessment methodologies utilizing data from short-term bioassays, therefore, are ...

  4. IN SITU BIOASSAY CHAMBER FOR ASSESSMENT OF SEDIMENT TOXICITY AND BIOACCUMULATION USING BENTHIC INVERTEBRATES

    EPA Science Inventory

    In this study, we describe the construction of a simple, inexpensive bioassay chamber for testing sediment toxicity (survival and growth) and bioaccumulation under field conditions using the midge Chironomus tentans and the oligochaete Lumbriculus variegatus. The test chamber is ...

  5. DEVELOPMENT OF BIOASSAY PROCEDURES FOR DEFINING POLLUTION OF HARBOR SEDIMENTS. PART I

    EPA Science Inventory

    This research investigates bioassay methods which may be useful in assessing the degree of pollution of harbor sediments. Procedures studied include 96 hr. toxicity tests employing Hexagenia limbata, Daphnia magna and Pontoporeia affinis as biological probes, monitoring cough fre...

  6. CHARACTERIZING THE GENOTOXICITY OF HAZARDOUS INDUSTRIAL WASTES AND EFFLUENTS USING SHORT-TERM BIOASSAYS

    EPA Science Inventory

    This chapter demonstrates that short-term bioassays can reliably and expeditiously measure the genotoxic potential of hazardous industrial wastes and effluents. etrochemical wastes have been studied in detail, especially discharges from chemical manufacturing plants and textile a...

  7. EVALUATION OF THE MUTAGENICITY AND CARCINOGENICITY OF MOTOR VEHICLE EMISSIONS IN SHORT-TERM BIOASSAYS

    EPA Science Inventory

    Incomplete combustion of fuel in motor vehicles results in the emission of submicron carbonaceous particles which, after cooling and dilution, contain varying quantities of extractable organic constituents. These organics are mutagenic in bacteria. Confirmatory bioassays in mamma...

  8. Comparative susceptibility of bemisia tabaci to imidacloprid in field- and laboratory-based bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bemisia tabaci biotype B is a resistance-prone pest of protected and open agriculture. Systemic uptake bioassays used in resistance monitoring programs have provided important information on susceptibility to neonicotinoid insecticides, but have remained decoupled from field performance. Simultaneou...

  9. EVALUATION OF THREE FISH SPECIES AS BIOASSAY ORGANISMS FOR DREDGED MATERIAL TESTING

    EPA Science Inventory

    Three fish species, Cyprinodon variegatus, Fundulus similis, and Menidia menidia, were evaluated to determine which is most suitable as a bioassay organism for solid phase testing of dredged material. Acute toxicity and bioaccumulation of polychlorinated biphenyls (PCBs) were mon...

  10. Harmonia Axyridis Adults Avoid Catnip and Grapefruit-derived Terpenoids in Laboratory Bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We observed the avoidance behavior of the multicolored Asian lady beetle, Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae), when adults were exposed to volatiles derived from catnip oil and grapefruit seed. In replicated laboratory bioassays, beetles avoided contact with volatiles emanating f...

  11. Evaluation of toxicity of selected insecticides against thrips on cotton in laboratory bioassays

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Adult vial technique (AVT) and spray table bioassays were conducted to evaluate toxicity of selected insecticides against immature and adult Western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). In AVT, technical insecticides comprising of organophosphates (d...

  12. 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

  13. Laboratory detection of extended-spectrum-beta-lactamase-producing Enterobacteriaceae: evaluation of two screening agar plates and two confirmation techniques.

    PubMed

    Overdevest, I T M A; Willemsen, I; Elberts, S; Verhulst, C; Kluytmans, J A J W

    2011-02-01

    The worldwide prevalence of extended-spectrum-beta-lactamase-producing ESBL-producing Enterobacteriaceae (ESBL-E) is increasing, making the need for optimized detection techniques more urgent. In this study we investigated the performance of two ESBL-E screening and two ESBL-E confirmation techniques. In accordance with the Dutch national guidelines (www.wip.nl), a collection of 642 highly resistant Enterobacteriaceae strains, as identified by Vitek2, was used to test the performances of two screening techniques (EbSA ESBL agar plate and ChromID ESBL agar plate) and of two confirmation techniques (MIC-strip ESBL and Vitek2 ESBL test panel). The individual test results were compared by using Etest, followed by a combination disk test if Etest results were inconclusive. Among group 1 isolates (Escherichia coli, Klebsiella spp., Proteus spp., Salmonella spp., and Shigella spp.) 291 (57.6%) were ESBL-E, versus 65 (47.4%) in group 2 (Enterobacter spp., Citrobacter spp., Morganella morganii, Serratia spp., and Providencia spp.). The sensitivities of all four tests for group 1 were comparable (EbSA, 96.6%; ChromID, 97.3%; MIC-strip, 99.6%; and Vitek2, 95.1%). The specificities of the EbSA and ChromID were the same (93.9%). However, the confirmation techniques produced many inconclusive test results, which reduces the applicability in routine laboratories. Only the two screening agar plates were validated for ESBL testing of group 2 microorganisms. They showed comparable sensitivities; however, the EbSA screening agar plate had a significantly higher specificity (78.6% versus 44.3%). In conclusion the screening agar plates performed better than the two confirmation techniques. The EbSA agar plate had the best overall performance. PMID:21123527

  14. A Standardized Lepidopteran Bioassay to Investigate the Bioactivity of Insecticidal Proteins Produced in Transgenic Crops.

    PubMed

    Graser, Gerson; Walters, Frederick S

    2016-01-01

    Insecticidal bioassays are the only reliable method to investigate the biological activity of an insecticidal protein and therefore provide an essential toolkit for the characterization and potency determination of these proteins. Here we present a standardized method for a lepidopteran larval bioassay, which is optimized to specifically estimate activity of insecticidal proteins produced in transgenic plants. The treatment can be either applied to the surface of the artificial diet, or blended into the diet. PMID:26614295

  15. Issues in weighting bioassay data for use in regressions for internal dose assessments

    SciTech Connect

    Strom, D.J.

    1992-11-01

    For use of bioassay data in internal dose assessment, research should be done to clarify the goal desired, the choice of method to achieve the goal, the selection of adjustable parameters, and on the ensemble of information that is available. Understanding of these issues should determine choices of weighting factors for bioassay data used in regression models. This paper provides an assessment of the relative importance of the various factors.

  16. A simple, rapid bioassay for detecting effects of pollutants on bacteria

    SciTech Connect

    Bauer, N.J.; Seidler, R.J.; Knittel, M.D.

    1981-12-01

    A screening bioassay needs to be rapid, and sensitive. The bioassay is described which is accurate, inexpensive, and which utilizes bacteria as the toxicity predictor. The basis of the test involves measuring the kinetics of dissolved oxygen depletion by a mixed microbial population following exposure to a pollutant and allows results to be obtained in as little as 40 min. Pollutants tested were cadmium, copper, nickel, sulfate, diuron, pentachlorophenol, atrazine, tricholoracetic acid, dimethylformamide, and diazinon. (JMT)

  17. Application of root bioassays to detect nutrient deficiencies in fast-growing trees and agroforestry crops

    SciTech Connect

    Harrison, A.F.; Dighton, J.; Jones, H.E.

    1992-12-31

    A new method for the detection of nutrient deficiencies is outlined and recommended as an alternative to conventional soil and foliar analyses. Bioassays are conducted to measure the uptake and supply of the macronutrients. Examples are quoted of the successful use of this technique with Eucalyptus and Sitka spruce. The bioassays have been shown to give equally good results with a range of tree and ground crops.

  18. Variation in the excitability of developed D. discoideum cells as a function of agar concentration in the substrate

    NASA Astrophysics Data System (ADS)

    Oikawa, Noriko; Bae, Albert; Amselem, Gabriel; Bodenschatz, Eberhard

    2010-03-01

    In the absence of nutrients, Dictyostelium discoideum cells enter a developmental cycle--they signal each other, aggregate, and ultimately form fruiting bodies. During the signaling stage, the cells relay waves of cyclic adenosine 3',5' monophosphate (cAMP). We observed a transition from spiral to circular patterns in the signaling wave, depending on the agar concentration of the substrate. In this talk we will present the changes in the times for the onset of signaling and synchronization versus agar concentration, as measured by spectral entropy. We also will discuss the origin of these effects.

  19. Modified agar dilution susceptibility testing method for determining in vitro activities of antifungal agents, including azole compounds.

    PubMed Central

    Yoshida, T; Jono, K; Okonogi, K

    1997-01-01

    In vitro activities of antifungal agents, including azole compounds, against yeasts were easily determined by using RPMI-1640 agar medium and by incubating the plates in the presence of 20% CO2. The end point of inhibition was clear by this method, even in the case of azole compounds, because of the almost complete inhibition of yeast growth at high concentrations which permitted weak growth of some Candida strains by traditional methods. MICs obtained by the agar dilution method were similar to those obtained by the broth dilution method proposed by the National Committee for Clinical Laboratory Standards. PMID:9174197

  20. In situ bioassay using Chironomus riparius: An intermediate between laboratory and field sediment quality assessments

    SciTech Connect

    Guchte, C. van de; Grootelaar, L.; Naber, A.

    1995-12-31

    Benthic macroinvertebrates like chironomid larvae are important indicators for sediment quality. Both in field surveys and laboratory bioassays effect parameters like abundance, survival, growth, larval development and morphological abnormalities of chironomids are recommended biological endpoints to assess the impact of sediment associated contaminants. Now and then results from field surveys on contaminated sites appeared to differ from results in laboratory bioassays on sediment field samples from the same sites. The impact of so-called modifying factors like temperature, oxygen levels and the availability of food could be studied in the laboratory. However, these factors could not fully explain the observed differences. In situ bioassays have been developed to bridge the gap between laboratory and field derived data with respect to the exposure of cultured Chironomus riparius larvae versus field collected Chironomus sp. larvae. Control survival in the in situ bioassays was within acceptable limits (> 80%). Effects observed during the caged exposure of laboratory cultured first instar larvae at contaminated sites were in agreement with the hypothesis that adequate in-field bioassessment reduces uncertainties inherent in the use of standardized laboratory bioassays. Although relative risk ranking of chemicals or contaminated sites can rely upon standard testing protocols, in situ bioassays can give a better insight in exposure-effect relationships under actual field conditions.

  1. An overview of the PubChem BioAssay resource.

    PubMed

    Wang, Yanli; Bolton, Evan; Dracheva, Svetlana; Karapetyan, Karen; Shoemaker, Benjamin A; Suzek, Tugba O; Wang, Jiyao; Xiao, Jewen; Zhang, Jian; Bryant, Stephen H

    2010-01-01

    The PubChem BioAssay database (http://pubchem.ncbi.nlm.nih.gov) is a public repository for biological activities of small molecules and small interfering RNAs (siRNAs) hosted by the US National Institutes of Health (NIH). It archives experimental descriptions of assays and biological test results and makes the information freely accessible to the public. A PubChem BioAssay data entry includes an assay description, a summary and detailed test results. Each assay record is linked to the molecular target, whenever possible, and is cross-referenced to other National Center for Biotechnology Information (NCBI) database records. 'Related BioAssays' are identified by examining the assay target relationship and activity profile of commonly tested compounds. A key goal of PubChem BioAssay is to make the biological activity information easily accessible through the NCBI information retrieval system-Entrez, and various web-based PubChem services. An integrated suite of data analysis tools are available to optimize the utility of the chemical structure and biological activity information within PubChem, enabling researchers to aggregate, compare and analyze biological test results contributed by multiple organizations. In this work, we describe the PubChem BioAssay database, including data model, bioassay deposition and utilities that PubChem provides for searching, downloading and analyzing the biological activity information contained therein. PMID:19933261

  2. Growth of clinical isolates of anaerobic bacteria on agar media: effects of media composition, storage conditions, and reduction under anaerobic conditions.

    PubMed Central

    Murray, P R

    1978-01-01

    The quantitative growth, the colony size, and the rate of growth of 47 clinical anaerobic isolates were compared on five different media, namely Brucella agar, brain heart infusion agar, Columbia agar, Schaedler agar, and tryptic soy agar. There was no significant difference in the quantitative growth of the anaerobes inoculated onto the five media. Although no single medium was superior for the growth of all isolates, 12 of 22 isolates, inoculated onto media stored for 4 weeks or less, grew best on Schaedler agar. The effects of supplementation of the media with reducing agents and reduction of the media before use were also analyzed and were found to be affected by the composition and length of storage of the media, as well as the bacteria tested. PMID:744801

  3. Use of benzimidazole agar plates to assess fall armyworm (Lepidoptera: Noctuidae) feeding on excised maize and sorghum leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is an economically significant pest of sorghum and maize. To screen sorghum and maize germplasm for resistance to fall armyworm feeding, field, greenhouse, or lab bioassays are often utilized individually or in combinatio...

  4. Defusing Diffusion

    ERIC Educational Resources Information Center

    Dou, Remy; Hogan, DaNel; Kossover, Mark; Spuck, Timothy; Young, Sarah

    2013-01-01

    Diffusion has often been taught in science courses as one of the primary ways by which molecules travel, particularly within organisms. For years, classroom teachers have used the same common demonstrations to illustrate this concept (e.g., placing drops of food coloring in a beaker of water). Most of the time, the main contributor to the motion…

  5. Relativistic diffusion

    NASA Astrophysics Data System (ADS)

    Haba, Z.

    2009-02-01

    We discuss relativistic diffusion in proper time in the approach of Schay (Ph.D. thesis, Princeton University, Princeton, NJ, 1961) and Dudley [Ark. Mat. 6, 241 (1965)]. We derive (Langevin) stochastic differential equations in various coordinates. We show that in some coordinates the stochastic differential equations become linear. We obtain momentum probability distribution in an explicit form. We discuss a relativistic particle diffusing in an external electromagnetic field. We solve the Langevin equations in the case of parallel electric and magnetic fields. We derive a kinetic equation for the evolution of the probability distribution. We discuss drag terms leading to an equilibrium distribution. The relativistic analog of the Ornstein-Uhlenbeck process is not unique. We show that if the drag comes from a diffusion approximation to the master equation then its form is strongly restricted. The drag leading to the Tsallis equilibrium distribution satisfies this restriction whereas the one of the Jüttner distribution does not. We show that any function of the relativistic energy can be the equilibrium distribution for a particle in a static electric field. A preliminary study of the time evolution with friction is presented. It is shown that the problem is equivalent to quantum mechanics of a particle moving on a hyperboloid with a potential determined by the drag. A relation to diffusions appearing in heavy ion collisions is briefly discussed.

  6. Relativistic diffusion.

    PubMed

    Haba, Z

    2009-02-01

    We discuss relativistic diffusion in proper time in the approach of Schay (Ph.D. thesis, Princeton University, Princeton, NJ, 1961) and Dudley [Ark. Mat. 6, 241 (1965)]. We derive (Langevin) stochastic differential equations in various coordinates. We show that in some coordinates the stochastic differential equations become linear. We obtain momentum probability distribution in an explicit form. We discuss a relativistic particle diffusing in an external electromagnetic field. We solve the Langevin equations in the case of parallel electric and magnetic fields. We derive a kinetic equation for the evolution of the probability distribution. We discuss drag terms leading to an equilibrium distribution. The relativistic analog of the Ornstein-Uhlenbeck process is not unique. We show that if the drag comes from a diffusion approximation to the master equation then its form is strongly restricted. The drag leading to the Tsallis equilibrium distribution satisfies this restriction whereas the one of the Jüttner distribution does not. We show that any function of the relativistic energy can be the equilibrium distribution for a particle in a static electric field. A preliminary study of the time evolution with friction is presented. It is shown that the problem is equivalent to quantum mechanics of a particle moving on a hyperboloid with a potential determined by the drag. A relation to diffusions appearing in heavy ion collisions is briefly discussed. PMID:19391727

  7. Bioassay of thermal protection afforded by candidate flight suit fabrics.

    PubMed

    Knox, F S; Wachtel, T L; McCahan, G R

    1979-10-01

    The United States Army Aeromedical Research Laboratory (USAARL) porcine cutaneous bioassay technique was used to determine what mitigating effect four thermally protective flight suit fabrics would have on fire-induced skin damage. The fabrics were 4.8-ox twill weave Nomex aramide, 4.5-oz stabilized twill weave polybenzimidazole, 4.8-oz plain weave experimental high-temperature polymer (HT4), and 4.8-oz plain weave Nomex aramide (New Weave Nomex or NWN). Each fabric sample was assayed 20 times in each of four configurations: as a single layer in contact with the skin; as a single layer with a 6.35 mm (0.25 in) air gap between fabric and skin; in conjuction with a cotton T-shirt with no air gaps; and, finally, in conjuction with a T-shirt with a 6.35 mm air gap between T-shirt and fabric. Bare skin was used as a control. A JP-4 fueled furnace was used as a thermal source and was adjested to deliver a mean heat flux of 3.07 cal/cm2/s. The duration of exposure was 5 s. Four hundred burn sites were graded using clinical observation and microscopic techniques. Used as single layers, none of the fabrics demonstrated superiority in providing clinically significant protection. When used with a cotton T-shirt, protection was improved. Protection improved progressively for all fabrics and configuration when an air gap was introduced. The experimental high-temperature polymer consistently demonstrated lower heat flux transmission in all configurations, but did not significantly reduce clinical burns. PMID:518445

  8. [Bioassay for enrich-blood bioactivity of Agelicae Sinensis Radix].

    PubMed

    Wang, Xiao-xiao; Zhang, Li-hong; Li, Xi; Wang, Ye; Rong, Zu-yuan; Wei, Hong-ping; Song, Qi-rui; Lv, Guang-hua

    2015-04-01

    Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality. PMID:26281565

  9. Plasmonically amplified bioassay - Total internal reflection fluorescence vs. epifluorescence geometry.

    PubMed

    Hageneder, Simone; Bauch, Martin; Dostalek, Jakub

    2016-08-15

    This paper investigates plasmonic amplification in two commonly used optical configurations for fluorescence readout of bioassays - epifluorescence (EPF) and total internal reflection fluorescence (TIRF). The plasmonic amplification in the EPF configuration was implemented by using crossed gold diffraction grating and Kretschmann geometry of attenuated total reflection method (ATR) was employed in the TIRF configuration. Identical assay, surface architecture for analyte capture, and optics for the excitation, collection and detection of emitted fluorescence light intensity were used in both TIRF and EPF configurations. Simulations predict that the crossed gold diffraction grating (EPF) can amplify the fluorescence signal by a factor of 10(2) by the combination of surface plasmon-enhanced excitation and directional surface plasmon-coupled emission in the red part of spectrum. This factor is about order of magnitude higher than that predicted for the Kretschmann geometry (TIRF) which only took advantage of the surface plasmon-enhanced excitation. When applied for the readout of sandwich interleukin 6 (IL-6) immunoassay, the plasmonically amplified EPF geometry designed for Alexa Fluor 647 labels offered 4-times higher fluorescence signal intensity compared to TIRF. Interestingly, both geometries allowed reaching the same detection limit of 0.4pM despite of the difference in the fluorescence signal enhancement. This is attributed to inherently lower background of fluorescence signal for TIRF geometry compared to that for EPF which compensates for the weaker fluorescence signal enhancement. The analysis of the inflammation biomarker IL-6 in serum at medically relevant concentrations and the utilization of plasmonic amplification for the fluorescence measurement of kinetics of surface affinity reactions are demonstrated for both EPF and TIRF readout. PMID:27260457

  10. Episodic acidification of small streams in the northeastern united states: Fish mortality in field bioassays

    USGS Publications Warehouse

    Van Sickle, J.; Baker, J.P.; Simonin, H.A.; Baldigo, Barry P.; Kretser, W.A.; Sharpe, W.E.

    1996-01-01

    In situ bioassays were performed as part of the Episodic Response Project, to evaluate the effects of episodic stream acidification on mortality of brook trout (Salvelinus fontinalis) and forage fish species. We report the results of 122 bioassays in 13 streams of the three study regions: the Adirondack mountains of New York, the Catskill mountains of New York, and the Northern Appalachian Plateau of Pennsylvania. Bioassays during acidic episodes had significantly higher mortality than did bioassays conducted under nonacidic conditions, but there was little difference in mortality rates in bioassays experiencing acidic episodes and those experiencing acidic conditions throughout the test period. Multiple logistic regression models were used to relate bioassay mortality rates to summary statistics of time-varying stream chemistry (inorganic monomeric aluminum, calcium, pH, and dissolved organic carbon) estimated for the 20-d bioassay periods. The large suite of candidate regressors also included biological, regional, and seasonal factors, as well as several statistics summarizing various features of aluminum exposure duration and magnitude. Regressor variable selection and model assessment were complicated by multicol-linearity and overdispersion. For the target fish species, brook trout, bioassay mortality was most closely related to time-weighted median inorganic aluminum. Median Ca and minimum pH offered additional explanatory power, as did stream-specific aluminum responses. Due to high multicollinearity, the relative importance of different aluminum exposure duration and magnitude variables was difficult to assess, but these variables taken together added no significant explanatory power to models already containing median aluminum. Between 59 and 79% of the variation in brook trout mortality was explained by models employing between one and five regressors. Simpler models were developed for smaller sets of bioassays that tested slimy and mottled sculpin

  11. [An oropharyngeal tularemia case diagnosed by the isolation of Francisella tularensis on human blood agar].

    PubMed

    Ozel, Gönül; Arslan, Ilker Burak; Yeşilyurt, Murat; Celebi, Bekir; Kılıç, Selçuk

    2010-10-01

    Tularemia which is a multisystem disease of humans and some animals, is endemic in North America, some parts of Europe and Asia. The causative agent, Francisella tularensis, is a fastidious gram-negative, intracellular bacterium which requires supplementation with sulphydryl compounds (cysteine, cystine, thiosulphate, isoVitaleX) for growth on common laboratory media. In this report, a case of oropharyngeal tularemia diagnosed by the isolation of the causative agent on non-selective-common microbiological agar, has been presented. The patient was from Yozgat located in central Anatolia where tularemia has not been reported so far. Forty-two years old male was admitted to the hospital with two weeks history of sudden onset fever, headache, generalized aches, sore throat, and cervical tender lump on the left. Physical examination revealed bilateral exudative tonsillitis and tender posterior cervical lymphadenopathy. He has been empirically treated with amoxicilin-clavulanic acid for 7 days with initial diagnosis of acute tonsillopharyngitis. However, he was admitted to the hospital since the symptoms persisted and swelling increased despite antibiotic therapy. Microscopical examination of the Gram and Ehrlich-Ziehl-Neelsen stained smears prepared from the surgically drained lymph node revealed PMNL, with no evidence of bacteria. Routine cultures of the lymph node material yielded growth of gram-negative coccobacilli only on human blood agar and the cultures were negative for pyogenic bacteria, acid-fast organisms and fungi. Pathologic examination of the drainage material revealed suppurative inflammation. Lymph node aspirate and serum samples of the patient together with the isolated strain were sent to reference laboratory for further investigation in accordance to the clinical and laboratory findings compatible with tularemia. The isolate was confirmed as F.tularensis by slide agglutination and direct immunofluorescence antibody tests, and identified as F

  12. 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

  13. Choline chloride based ionic liquid analogues as tool for the fabrication of agar films with improved mechanical properties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the present paper, we test the suitability of Choline-Cl/urea (DES-U) and Choline-Cl/glycerol (DES-G) eutectic mixtures at 1:2 molar ratios for the production of agar biodegradable films. A three-step process is proposed: pre-solubilization of polymer in DES followed by compression-molding and s...

  14. Controlled evaluation of the agar-slide and radiometric blood culture systems for the detection of bacteremia and fungemia.

    PubMed Central

    Weinstein, M P; Reller, L B; Mirrett, S; Stratton, C W; Reimer, L G; Wang, W L

    1986-01-01

    A commercially available agar-slide blood culture bottle (Septi-Chek; Roche Diagnostics, Div. Hoffman-La Roche, Inc., Nutley, N.J.) was compared with the radiometric blood culture system (BACTEC; Johnston Laboratories, Inc., Towson, Md.) in 8,544 paired blood cultures from adult patients. The systems were inoculated with equal volumes (10 ml) of blood. Overall, there was no statistically significant difference between the two systems in the recovery of clinically important microorganisms, but significantly more members of the family Enterobacteriaceae other than Escherichia coli were detected by the agar-slide system (P less than 0.005). The agar-slide system detected more fungi, and the BACTEC detected more anaerobic bacteria; however, small numbers of recovered organisms precluded statistical significance. When microorganisms grew in both systems, their presence was detected one or more days earlier in the BACTEC (P less than 0.001). More contaminants grew in the agar-slide system (P less than 0.001). Both systems performed well, and either system should provide high yield and prompt detection of positive blood cultures in patients with bacteremia and fungemia if used in an optimal way as recommended by the respective manufacturers. PMID:3517047

  15. Effects of immersion disinfection of agar-alginate combined impressions on the surface properties of stone casts.

    PubMed

    Iwasaki, Yukiko; Hiraguchi, Hisako; Iwasaki, Eriko; Yoneyama, Takayuki

    2016-01-01

    This study investigated the effects of disinfection of agar-alginate combined impressions on the surface properties of the resulting stone casts. Two brands of cartridge-form agar impression material and one alginate impression material were used. Agar-alginate combined impressions of smooth glass plates were prepared. The impressions were immersed in 0.55% ortho-phthalaldehyde solution or 0.5% sodium hypochlorite solution for 1, 3, 5 and 10 min. A stone cast made with an impression that had not been immersed was prepared as a control. The surface roughness (Ra) of the stone casts was measured, and the cast surfaces were observed by SEM. Immersion of agar-alginate combined impressions in 0.5% sodium hypochlorite solution for up to 10 min had no serious adverse effects on the surface properties of the stone casts. In contrast, even 1 min of immersion in 0.55% ortho-phthalaldehyde solution caused deterioration of the cast surface properties. PMID:26843442

  16. Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells.

    PubMed

    Horibata, Sachi; Vo, Tommy V; Subramanian, Venkataraman; Thompson, Paul R; Coonrod, Scott A

    2015-01-01

    Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation assay (hereafter called soft agar assay), which measures the ability of cells to proliferate in semi-solid matrices, remain a hallmark of cancer research. A key advantage of this technique over conventional 2D monolayer or 3D spheroid cell culture assays is the close mimicry of the 3D cellular environment to that seen in vivo. Importantly, the soft agar assay also provides an ideal tool to rigorously test the effects of novel compounds or treatment conditions on cell proliferation and migration. Additionally, this assay enables the quantitative assessment of cell transformation potential within the context of genetic perturbations. We recently identified peptidylarginine deiminase 2 (PADI2) as a potential breast cancer biomarker and therapeutic target. Here we highlight the utility of the soft agar assay for preclinical anti-cancer studies by testing the effects of the PADI inhibitor, BB-Cl-amidine (BB-CLA), on the tumorigenicity of human ductal carcinoma in situ (MCF10DCIS) cells. PMID:26067809

  17. Hydrogen-bond-mediated in situ fabrication of AgNPs/agar/PAN electrospun nanofibers as reproducible SERS substrates.

    PubMed

    Yang, Tong; Yang, Hui; Zhen, Shu Jun; Huang, Cheng Zhi

    2015-01-28

    Reproducibility in surface enhanced Raman scattering (SERS) measurements is a challenge. This work developed a facile way to make highly dispersed uniform silver nanoparticles (AgNPs) loaded in the agar/polyacrylonitrile (PAN) nanofibers by the coupling the electrospinning technology from metal complex-containing polymer solution and in situ photoreductive technique. Agar, as hydrophilic component, was introduced into the electrospinning solution considering that its abundant hydroxyl group sites could greatly improve the contents of silver ions in the polymers because of the rich silver ion chelated with the hydroxyl group, whereas hydrophilic agar was integrated with hydrophobic PAN by -OH···N≡C- hydrogen bonds as a bridge. Meanwhile, the in situ photoreductive reaction was made under different light irradiations such as desk lamp, 365 nm UV-lamp, and 254 nm UV-lamp. High yield of stable AgNPs with highly uniform and dispersion are available in the agar/PAN nanofibers after the in situ photoreductive reaction, supplying the possibility of reproducible SERS signals. To identify that concept of proof, a facile approach for the determination of malachite green (MG) in three environmental practical samples was demonstrated by using the composite nanofibrous material irradiated by 365 nm UV-lamp, giving the minimum detection concentration of MG as low as 0.1 μmol/L with a good linear response ranging from 0.1-100 μmol/L (R(2) = 0.9960). PMID:25546719

  18. NMR analysis of weak molecular interactions using slice-selective experiments via study of concentration gradients in agar gels.

    PubMed

    Mitrev, Y; Simova, S; Jeannerat, D

    2016-04-01

    Weak molecular interactions can be localized and quantified using a single NMR experiment analysing concentration gradients generated in agar gels. The spectra from various cross-sections along the gradient were obtained using a slice-selective pulse sequence realisable with standard NMR equipment. PMID:27009847

  19. The Limits of Two-Year Bioassay Exposure Regimens for Identifying Chemical Carcinogens

    PubMed Central

    Huff, James; Jacobson, Michael F.; Davis, Devra Lee

    2008-01-01

    Background Chemical carcinogenesis bioassays in animals have long been recognized and accepted as valid predictors of potential cancer hazards to humans. Most rodent bioassays begin several weeks after birth and expose animals to chemicals or other substances, including workplace and environmental pollutants, for 2 years. New findings indicate the need to extend the timing and duration of exposures used in the rodent bioassay. Objectives In this Commentary, we propose that the sensitivity of chemical carcinogenesis bio-assays would be enhanced by exposing rodents beginning in utero and continuing for 30 months (130 weeks) or until their natural deaths at up to about 3 years. Discussion Studies of three chemicals of different structures and uses—aspartame, cadmium, and toluene—suggest that exposing experimental animals in utero and continuing exposure for 30 months or until their natural deaths increase the sensitivity of bioassays, avoid false-negative results, and strengthen the value and validity of results for regulatory agencies. Conclusions Government agencies, drug companies, and the chemical industry should conduct and compare the results of 2-year bioassays of known carcinogens or chemicals for which there is equivocal evidence of carcinogenicity with longer-term studies, with and without in utero exposure. If studies longer than 2 years and/or with in utero exposure are found to better identify potential human carcinogens, then regulatory agencies should promptly revise their testing guidelines, which were established in the 1960s and early 1970s. Changing the timing and dosing of the animal bioassay would enhance protection of workers and consumers who are exposed to potentially dangerous workplace or home contaminants, pollutants, drugs, food additives, and other chemicals throughout their lives. PMID:19057693

  20. Benchmarking organic micropollutants in wastewater, recycled water and drinking water with in vitro bioassays.

    PubMed

    Escher, Beate I; Allinson, Mayumi; Altenburger, Rolf; Bain, Peter A; Balaguer, Patrick; Busch, Wibke; Crago, Jordan; Denslow, Nancy D; Dopp, Elke; Hilscherova, Klara; Humpage, Andrew R; Kumar, Anu; Grimaldi, Marina; Jayasinghe, B Sumith; Jarosova, Barbora; Jia, Ai; Makarov, Sergei; Maruya, Keith A; Medvedev, Alex; Mehinto, Alvine C; Mendez, Jamie E; Poulsen, Anita; Prochazka, Erik; Richard, Jessica; Schifferli, Andrea; Schlenk, Daniel; Scholz, Stefan; Shiraishi, Fujio; Snyder, Shane; Su, Guanyong; Tang, Janet Y M; van der Burg, Bart; van der Linden, Sander C; Werner, Inge; Westerheide, Sandy D; Wong, Chris K C; Yang, Min; Yeung, Bonnie H Y; Zhang, Xiaowei; Leusch, Frederic D L

    2014-01-01

    Thousands of organic micropollutants and their transformation products occur in water. Although often present at low concentrations, individual compounds contribute to mixture effects. Cell-based bioassays that target health-relevant biological endpoints may therefore complement chemical analysis for water quality assessment. The objective of this study was to evaluate cell-based bioassays for their suitability to benchmark water quality and to assess efficacy of water treatment processes. The selected bioassays cover relevant steps in the toxicity pathways including induction of xenobiotic metabolism, specific and reactive modes of toxic action, activation of adaptive stress response pathways and system responses. Twenty laboratories applied 103 unique in vitro bioassays to a common set of 10 water samples collected in Australia, including wastewater treatment plant effluent, two types of recycled water (reverse osmosis and ozonation/activated carbon filtration), stormwater, surface water, and drinking water. Sixty-five bioassays (63%) showed positive results in at least one sample, typically in wastewater treatment plant effluent, and only five (5%) were positive in the control (ultrapure water). Each water type had a characteristic bioanalytical profile with particular groups of toxicity pathways either consistently responsive or not responsive across test systems. The most responsive health-relevant endpoints were related to xenobiotic metabolism (pregnane X and aryl hydrocarbon receptors), hormone-mediated modes of action (mainly related to the estrogen, glucocorticoid, and antiandrogen activities), reactive modes of action (genotoxicity) and adaptive stress response pathway (oxidative stress response). This study has demonstrated that selected cell-based bioassays are suitable to benchmark water quality and it is recommended to use a purpose-tailored panel of bioassays for routine monitoring. PMID:24369993

  1. Experiences with Non-traditional Bioassay Methods in a Plutonium Processing Line

    SciTech Connect

    La Bone, T.R.

    2003-10-17

    An incident in an Savannah River Site (SRS) plutonium processing line (FB-Line) in 1999 highlighted the fact insoluble forms of plutonium exist at SRS that may not be readily monitored with the routine bioassay programs traditionally used at this site. To address this issue, a study was conducted in FB-Line with 21 participants for a year ending in July 2002. The purpose of the study was to examine the use of three non-traditional monitoring methods and, based on this experience, recommend a routine bioassay program that is capable of monitoring workers potentially exposed to insoluble plutonium. These non-traditional monitoring methods are personal air sampling (PAS), thermal ionization mass spectrometry (TIMS) of urine samples, and routine fecal bioassay. The main conclusions and recommendations of the study are: (1) A routine TIMS urine bioassay program, which is called the enhanced bioassay program (EBP), is recommended for workers in SRS facilities that have a reasonable potential for exposure to insoluble forms of plutonium. (2) Under certain conditions the EBP could result in onerous work restrictions. A contingency plan involving the use of PAS is recommended in this case. PAS is also recommended for workers who have had historic intakes of plutonium that interfere with the detection and interpretation of future intakes of insoluble plutonium. (3) For the EBP to be successful it must be used only for those workers who have a reasonable potential for exposure to insoluble plutonium, and these workers must take all necessary precautions to avoid cross-contamination of the urine (and follow-up fecal) samples. (4) Fecal bioassay is an important tool for follow-up to abnormal events, but routine fecal bioassay is not recommended. (5) The PAS data clearly shows that workers are exposed to low levels of airborne plutonium, but the participants appear to be unlikely to exceed a committed effective dose equivalent of 100 mrem from these exposures.

  2. [Variations in hyperbilirrubinemia in low birth weight newborns under phototherapy and continous or discontinous agar oral administration (author's transl)].

    PubMed

    Colomer, J; Moya, M; Marco, V; De Paredes, C; Escrivá, F; Vila, R

    1975-06-01

    Therapeutic attitude in hyperbilirrubinemia is always worth because other infrequent complications but not for this, less important. Phototherapy innocuousness, largely demonstrated, fosters its profilactic use at beginning and not only for those babies with serum bilirrubin over 10 mg % in the first day of life. Previously we have reported positive results with agar oral administration without collateral effects. On this grounds we have planned the following experience in a homogenous group of L.B.W.: one group was fed with agar previously to each formula administration; other group received the same amount of agar but divided in only three administrations in 24 hours; the last group received continuous phototherapy for 96 hours with a white cold fluorescent light from a source of 8-Vita-lite lamp of 40 watts with a intensity of 500 foot candle and 30 lumens. All of these babies weighed less than 2.500 g. and were between 10 and 90 percentil of Lubschenko diagram. They were fed with the same formula and same time table with no infusions, rejecting all that presented any type of pathology. Obstetric conditions were basically identical. This population was randomly divided in four groups. 1) Control group with no profilaxis, but with identical bilirrubin andhematocrit determinations. 2) Group with continuous agar oral administration, 125 mg. before each of the seven formula feeding. 3) Group with discontinuous agar administration, 250 mg. before three of the seven formula feeding. 4) Group with continuous phototherapy for 96 hours. These is initial identification of the groups with statistic signification, and after that a quantitative and sequential evolution of bilirrubin is analized in each group. PMID:1155873

  3. Theoretical and experimental NMR studies on muscimol from fly agaric mushroom (Amanita muscaria)

    NASA Astrophysics Data System (ADS)

    Kupka, Teobald; Wieczorek, Piotr P.

    2016-01-01

    In this article we report results of combined theoretical and experimental NMR studies on muscimol, the bioactive alkaloid from fly agaric mushroom (Amanita muscaria). The assignment of 1H and 13C NMR spectra of muscimol in DMSO-d6 was supported by additional two-dimensional heteronuclear correlated spectra (2D NMR) and gauge independent atomic orbital (GIAO) NMR calculations using density functional theory (DFT). The effect of solvent in theoretical calculations was included via polarized continuum model (PCM) and the hybrid three-parameter B3LYP density functional in combination with 6-311++G(3df,2pd) basis set enabled calculation of reliable structures of non-ionized (neutral) molecule and its NH and zwitterionic forms in the gas phase, chloroform, DMSO and water. GIAO NMR calculations, using equilibrium and rovibrationally averaged geometry, at B3LYP/6-31G* and B3LYP/aug-cc-pVTZ-J levels of theory provided muscimol nuclear magnetic shieldings. The theoretical proton and carbon chemical shifts were critically compared with experimental NMR spectra measured in DMSO. Our results provide useful information on its structure in solution. We believe that such data could improve the understanding of basic features of muscimol at atomistic level and provide another tool in studies related to GABA analogs.

  4. Fungistatic activity of flaxseed in potato dextrose agar and a fresh noodle system.

    PubMed

    Xu, Yingying; Hall, Clifford; Wolf-Hall, Charlene; Manthey, Frank

    2008-02-10

    Although numerous researchers have studied flaxseed as a food ingredient for its health benefits, flaxseed (Linum usitatissimum) has never been considered as a food preservative. The objective of this study was to investigate the effect of flaxseed flour (FF) concentration (0, 6, 9, 12, and 15% wt/wt), cultivar ('Omega' and brown) and source (four seed companies located in Minnesota and North Dakota) on flaxseed fungistatic activity. Fungal radial growth was used to assess the fungistatic activity of FF in both potato dextrose agar (PDA) medium and a fresh noodle system. Strains of Penicillium chrysogenum, Aspergillus flavus, Fusarium graminearum, and a Penicillium sp. isolated from molded noodles were used as the test microorganisms. Results showed that growth of F. graminearum was completely inhibited at all FF concentrations in PDA, and the inhibition of the other three test microorganisms increased with increasing FF concentrations. In the model noodle system, FF concentration at 9% or higher significantly reduced the mold count of fresh noodle during storage. In the inoculated noodle system, 6% FF addition was sufficient to significantly inhibit the growth of F. graminearum and A. flavus, whereas 9% FF concentrations showed fungistatic activity against P. chrysogenum and the Penicillium sp. isolate. Differences in the degree of mold inhibition were found among FFs obtained from different sources and cultivars. Results suggested that flaxseed possesses fungistatic activity and could be used as a multifunctional food ingredient. PMID:18077042

  5. Spectral filtering for improved pulsed photothermal temperature profiling in agar tissue phantoms

    PubMed Central

    Milanič, Matija; Majaron, Boris; Nelson, J. Stuart

    2009-01-01

    We present a systematic experimental comparison of pulsed photothermal temperature profiling utilizing the customary spectral band of the InSb radiation detector (λ=3.0 to 5.6 μm) and a narrowed acquisition band (4.5 to 5.6 μm). We use custom tissue phantoms composed of agar gel layers separated by thin absorbing layers. The laser-induced temperature profiles are reconstructed within the customary monochromatic approximation, using a custom minimization algorithm. In a detailed numerical simulation of the experimental procedure, we consider several acquisition spectral bands with the lower wavelength limit varied between 3.0 and 5.0 μm (imitating application of different long-pass filters). The simulated PPTR signals contain noise with amplitude and spectral characteristics consistent with our experimental system. Both experimental and numerical results indicate that spectral filtering reduces reconstruction error and broadening of temperature peaks, especially for shallower and more complex absorbing structures. For the simulated PPTR system and watery tissues, numerical results indicate an optimal lower wavelength limit of 3.8 to 4.2 μm. PMID:19123649

  6. Susceptibilities of genital mycoplasmas to the newer quinolones as determined by the agar dilution method.

    PubMed Central

    Kenny, G E; Hooton, T M; Roberts, M C; Cartwright, F D; Hoyt, J

    1989-01-01

    The increasing resistance of genital mycoplasmas to tetracycline poses a problem because tetracycline is one of the few antimicrobial agents active against Mycoplasma hominis, Ureaplasma urealyticum, chlamydiae, gonococci, and other agents of genitourinary-tract disease. Since the quinolones are a promising group of antimicrobial agents, the susceptibilities of M. hominis and U. urealyticum to the newer 6-fluoroquinolones were determined by the agar dilution method. Ciprofloxacin, difloxacin, and ofloxacin had good activity against M. hominis, with the MIC for 50% of isolates tested (MIC50) being 1 microgram/ml. Fleroxacin, lomefloxacin, pefloxacin, and rosoxacin had MIC50s of 2 micrograms/ml. Enoxacin, norfloxacin, and amifloxacin had MIC50s of 8 to 16 micrograms/ml, and cinoxacin and nalidixic acid were inactive (MIC50, greater than or equal to 256 micrograms/ml). Overall, the activities of 6-fluoroquinolones for ureaplasmas were similar to those for M. hominis, with MICs being the same or twofold greater. The most active 6-fluoroquinolones against ureaplasmas were difloxacin, ofloxacin, and pefloxacin, with MIC50s of 1 to 2 micrograms/ml. Ciprofloxacin was unusual in that the MIC50 for M. hominis was 1 microgram/ml, whereas the MIC50 for ureaplasmas was 8 micrograms/ml. Since the MIC50s for the most active quinolones approximate achievable concentrations in blood and urine, quinolones have promise in treating mycoplasmal infections. PMID:2712541

  7. National Committee for Clinical Laboratory Standards agar dilution susceptibility testing of anaerobic gram-negative bacteria.

    PubMed Central

    Brown, W J

    1988-01-01

    One hundred nine recent clinical isolates of anaerobic gram-negative bacteria were tested in triplicate by the National Committee for Clinical Laboratory Standards agar dilution procedure for their susceptibility to 32 antimicrobial agents. All isolates were inhibited by imipenem, but there were significant numbers of strains resistant to other beta-lactam drugs, and therefore the in vitro response to these antimicrobial agents cannot be predicted. This was particularly true for the bile-resistant or Bacteroides fragilis group. beta-Lactamase production was detected in 82% of the bacteroides with the nitrocefin test. Clavulanic acid combined with amoxicillin and ticarcillin and sulbactam combined with ampicillin resulted in synergistic activity against all beta-lactamase-positive organisms. Ceftizoxime was the most active of the cephalosporins. Two percent of the isolates were resistant to chloramphenicol and metronidazole. Clindamycin resistance was detected in 38% of the B. fragilis group, which is a marked increase from the 4% detected 10 years ago at this institution. PMID:3364956

  8. Theoretical and experimental NMR studies on muscimol from fly agaric mushroom (Amanita muscaria).

    PubMed

    Kupka, Teobald; Wieczorek, Piotr P

    2016-01-15

    In this article we report results of combined theoretical and experimental NMR studies on muscimol, the bioactive alkaloid from fly agaric mushroom (Amanita muscaria). The assignment of (1)H and (13)C NMR spectra of muscimol in DMSO-d6 was supported by additional two-dimensional heteronuclear correlated spectra (2D NMR) and gauge independent atomic orbital (GIAO) NMR calculations using density functional theory (DFT). The effect of solvent in theoretical calculations was included via polarized continuum model (PCM) and the hybrid three-parameter B3LYP density functional in combination with 6-311++G(3df,2pd) basis set enabled calculation of reliable structures of non-ionized (neutral) molecule and its NH and zwitterionic forms in the gas phase, chloroform, DMSO and water. GIAO NMR calculations, using equilibrium and rovibrationally averaged geometry, at B3LYP/6-31G* and B3LYP/aug-cc-pVTZ-J levels of theory provided muscimol nuclear magnetic shieldings. The theoretical proton and carbon chemical shifts were critically compared with experimental NMR spectra measured in DMSO. Our results provide useful information on its structure in solution. We believe that such data could improve the understanding of basic features of muscimol at atomistic level and provide another tool in studies related to GABA analogs. PMID:26312739

  9. Mercury and its bioconcentration factors in fly agaric (Amanita muscaria) from spatially distant sites in Poland.

    PubMed

    Falandysz, J; Lipka, K; Mazur, A

    2007-09-01

    Total mercury content has been determined in the fruiting bodies of fly agaric (Amanita muscaria) and topsoil layer (0-10 cm) collected from 14 spatially distant sites across Poland. Mercury was measured by cold-vapor atomic absorption spectroscopy (CV-AAS) after nitric acid (mushrooms) or nitric acid and sulfuric acid (soil) digestion of the samples. The caps, depending on the site, contained total mercury at mean concentrations from 0.24+/-0.13 to 1.4+/-0.6 microg/g dm (median 0.19-1.4 microg/g dm), and stalks from 0.18+/-0.06 to 0.71+/-0.26 microg/g dm (median 0.18-0.67 microg/g dm). An overall-mean the total mercury content for 204 caps and stalks was, respectively, 0.73+/-0.55 (0.05-3.3 microg/g dm) and 0.43+/-0.33 (0.09-2.3 microg/g dm). PMID:17849304

  10. Comparison of CHROMagar Salmonella Medium and Hektoen Enteric Agar for Isolation of Salmonellae from Stool Samples

    PubMed Central

    Gaillot, Olivier; Di Camillo, Patrick; Berche, Patrick; Courcol, René; Savage, Colette

    1999-01-01

    CHROMagar Salmonella (CAS), a new chromogenic medium, was retrospectively compared to Hektoen enteric agar (HEA) with 501 Salmonella stock isolates and was then prospectively compared to HEA for the detection and presumptive identification of Salmonella spp. with 508 stool samples before and after enrichment. All stock cultures (100%), including cultures of H2S-negative isolates, yielded typical mauve colonies on CAS, while 497 (99%) isolates produced typical lactose-negative, black-centered colonies on HEA. Following overnight incubation at 37°C, a total of 20 Salmonella strains were isolated from the 508 clinical samples. Sensitivities for primary plating and after enrichment were 95% (19 isolates) and 100% (20 isolates), respectively, for CAS and 80% (16 isolates) and 100% (20 isolates), respectively, for HEA. The specificity of CAS (88.9%) was significantly higher than that of HEA (78.5%; P < 0.0001). On the basis of its good sensitivity and specificity, CAS medium can be recommended for use for primary plating when human stool samples are screened for Salmonella spp. PMID:9986847

  11. Chemosensitivity measurements of human tumour cells by soft agar assays are influenced by the culture conditions.

    PubMed Central

    Endresen, L.; Tveit, K. M.; Rugstad, H. E.; Pihl, A.

    1985-01-01

    To investigate the influence of culture conditions on the in vitro responses of tumour cells to anticancer drugs, the sensitivities observed with the soft agar methods of Hamburger & Salmon (1977) (H-S) and of Courtenay & Mills (1978) (C-M) were compared. In all cases the ID50 values were determined from dose-response curves. Six human tumour cell lines exposed to 10 different agents, and 9 patients' melanomas exposed to 5 different agents, were examined. In the studies of cell lines the H-S method gave higher sensitivity values than the C-M method in 38 out of 52 cases, whereas in 14 cases the results were the same. In the patients' tumours the H-S method gave higher sensitivity in 21 of 35 cases, equal sensitivity in 11, and lower sensitivity in 3 cases. In many instances the ID50 values obtained with the two test systems differed by factors of 10 or more, both in the case of cell lines and tumour specimens. Systematic alterations in the culture conditions indicated that the presence or absence of rat erythrocytes is the most important factor responsible for the differences observed. Also, other factors, such as supplements (in the H-S method) and the use of different serum types, appeared to influence both colony growth and chemosensitivity. PMID:4005141

  12. Studies on the inactivation of medically important Candida species on agar surfaces using pulsed light.

    PubMed

    Farrell, Hugh; Garvey, Mary; Rowan, Neil

    2009-09-01

    Development of a pulsed-light (PL) approach to inanimate surface decontamination is timely, as the incidence of yeast-related infections in healthcare remains unacceptably high. Critical electrical and biological factors governing the efficacy of PL for the in vitro inactivation of medically important yeast were established in this study. Predetermined cell numbers of yeast were inoculated separately on agar plates and were flashed with < or =90 pulses of broad-spectrum light under varying operating conditions, and their inactivation was measured. Significant differences in inactivation among different yeasts occurred depending on the intensity of the applied lamp discharge energy and the amount of pulsing applied. Levels of yeast sensitivity also varied depending on the distance between the light source and the treatment surface used, and the population size, type and age of cultures treated. Yeast strains were shown to be significantly more resistant to PL irradiation compared with similarly treated bacterial control cultures. A clear relationship was observed between the concentration of eluted proteins from treated yeast and the severity of PL conditions, with scanning electron micrographs showing irreversible cellular damage. Therefore, the findings from this study will enable further development and optimization of PL as a method of decontaminating surfaces in healthcare setting. PMID:19624750

  13. Diffusion bonding

    DOEpatents

    Anderson, Robert C.

    1976-06-22

    1. A method for joining beryllium to beryllium by diffusion bonding, comprising the steps of coating at least one surface portion of at least two beryllium pieces with nickel, positioning a coated surface portion in a contiguous relationship with an other surface portion, subjecting the contiguously disposed surface portions to an environment having an atmosphere at a pressure lower than ambient pressure, applying a force upon the beryllium pieces for causing the contiguous surface portions to abut against each other, heating the contiguous surface portions to a maximum temperature less than the melting temperature of the beryllium, substantially uniformly decreasing the applied force while increasing the temperature after attaining a temperature substantially above room temperature, and maintaining a portion of the applied force at a temperature corresponding to about maximum temperature for a duration sufficient to effect the diffusion bond between the contiguous surface portions.

  14. Characterizing an agar/gelatin phantom for image guided dosing and feedback control of high-intensity focused ultrasound.

    PubMed

    Dunmire, Barbrina; Kucewicz, John C; Mitchell, Stuart B; Crum, Lawrence A; Sekins, K Michael

    2013-02-01

    The temperature dependence of an agar/gelatin phantom was evaluated. The purpose was to predict the material property response to high-intensity focused ultrasound (HIFU) for developing ultrasound guided dosing and targeting feedback. Changes in attenuation, sound speed, shear modulus and thermal properties with temperature were examined from 20°C to 70°C for 3 weeks post-manufacture. The attenuation decreased with temperature by a power factor of 0.15. Thermal conductivity, diffusivity and specific heat all increased linearly with temperature for a total change of approximately 16%, 10% and 6%, respectively. Sound speed had a parabolic dependence on temperature similar to that of water. Initially, the shear modulus irreversibly declined with even a slight increase in temperature. Over time, the gel maintained its room temperature shear modulus with moderate heating. A stable phantom was achieved within 2 weeks post-manufacture that possessed quasi-reversible material properties up to nearly 55°C. PMID:23245823

  15. Reading disc-based bioassays with standard computer drives.

    PubMed

    Yu, Hua-Zhong; Li, Yunchao; Ou, Lily M-L

    2013-02-19

    -based bioassays quantitatively. In this Account, we first provide a brief introduction to CD-related materials chemistry and microfluidics research. Then we describe the mild chemistry developed in our laboratory for the preparation of computer-readable biomolecular screening assays: photochemical activation of the polycarbonate (PC) disc surface and immobilization and delivery of probe and target biomolecules. We thoroughly discuss the analysis of the molecular recognition events: researchers can "read" these devices quantitatively with an unmodified optical drive of any personal computer. Finally, and critically, we illustrate our digitized molecular diagnosis approach with three trial systems: DNA hybridization, antibody-antigen binding, and ultrasensitive lead detection with a DNAzyme assay. These examples demonstrate the broad potential of this new analytical/diagnostic tool for medical screening, on-site food/water safety testing, and remote environmental monitoring. PMID:23025412

  16. The usefulness of a sediment bioassay with the gastropod Nassarius reticulatus in tributyltin monitoring programs.

    PubMed

    Laranjeiro, Filipe; Pérez, Sara; Navarro, Patricia; Carrero, José Antonio; Beiras, Ricardo

    2015-11-01

    Despite the use of tributyltin (TBT) had been banned worldwide in 2008 there is still evidence of its deleterious presence in environment. We evaluate the usefulness of a 28days sediment bioassay with Nassarius reticulatus females to monitor TBT pollution, using imposex as endpoint. In addition, butyltins were determined in sediments and tissues, and, whenever posible, imposex was assessed in native N. reticulatus at the same sites where sediments were sampled. In the bioassay, a significant increase in imposex parameters was obtained with three sediments (Vi2, Vi3, and Vi4). No correlation was found between this and TBT concentrations in sediment although good correlations were obtained for TBT in tissues, putting in evidence TBT bioavailability in sediment. A significant decrease in imposex from 2008 to 2013 in native snails was only observed at sites that did not cause any effect in the bioassay. In contrast, imposex levels in 2013 were kept as high as 2008 in one of the sites where a significant imposex increase in the bioassay was observed. The bioassay proves thus to be a practical and ecological relevant tool, as: (i) it can be conducted in sites with no native populations of snails, (ii) it provides early identification of polluted sites, anticipating future imposex levels or early identification of recovering, and (iii) it yields information on the bioavailable fraction of the TBT in the sediment. Therefore, this tool can be of extreme usefulness under the scope of recent European legislative frameworks. PMID:26318117

  17. Rainbow trout cell bioassay-derived relative potencies for halogenated aromatic hydrocarbons: Comparison and sensitivity analysis

    SciTech Connect

    Villeneuve, D.L.; Blankenship, A.L.; Giesy, J.P.; Richter, C.A.

    1999-05-01

    Rainbow trout hepatoma cells, stably transfected with a luciferase reporter gene under control of dioxin-responsive elements (RLT 2.0 cells) were used to derive relative potencies (RPs) for a variety of halogenated aromatic hydrocarbons (HAHs) that are structurally similar to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). This in vitro bioassay utilizes 96-well microplates, which provide high sample throughput and assay efficiency without affecting sensitivity. The RLT 2.0-derived potencies for dioxin and furan congeners, relative to 2,3,7,8-TCDD, ranged from 0.917 for 1,2,3,4,7,8-hexachlorodibenzofuran to 0.208 or 1,2,3,7,8-pentachlorodibenzofuran. All mono- and di-ortho polychlorinated biphenyls (PCBs) tested had RPs that were orders of magnitude less than TCDD, but point estimates could not be determined. The RLT 2.0-derived RPs were found to be comparable to both other rainbow trout-specific RPs and RPs based on mammalian bioassays. Sensitivity analysis suggested that the range of uncertainty associated with TCDD equivalent (TEQ) estimates based on RLT 2.0-derived RPs is approximately 10-fold. Within this degree of uncertainty and the context of this study, the RLT 2.0 bioassay showed no definitive biases or inaccuracies relative to similar mammalian- or fish-specific in vitro bioassays. Thus, the RLT 2.0 bioassay appears to be a useful tool for evaluating dioxin-like potency of HAHs to fish.

  18. Selecting a sensitive battery of bioassays to detect toxic effects of metals in effluents.

    PubMed

    de Paiva Magalhães, Danielly; da Costa Marques, Mônica Regina; Fernandes Baptista, Darcilio; Forsin Buss, Daniel

    2014-09-01

    The use of bioassay batteries is necessary to evaluate toxic effects at various biological levels. The selection of bioassays without prior testing and determination of the most sensitive/suitable groups for each impact may allow the discharge of effluents that pose a threat to the environment. The present study tested and selected a battery of sensitive ecotoxicological bioassays for detecting toxic effects of metals. The sensitivities of six organisms were evaluated (algae Pseudokirchneriella subcapitata and Chlorella vulgaris, Cladocera Daphnia similis and Ceriodaphnia dubia, and fish Poecilia reticulata and Danio rerio) after exposure to 10 individual metal species deemed toxic to the aquatic environment (Ag(+), Cd(2+), Cu(+), Cu(2+), Cr(3+), Cr(6+), Pb(2+), Ni(2+), Zn(2+), and Hg(2+)) and to real (steel-mill) and laboratory simulated effluents. In the bioassays, fish were the least sensitive; D. rerio showed no sensitivity to any of the effluents tested. P. subcapitata was a good bioindicator of Cr(3+) toxicity, and D. similis was the most sensitive organism to Hg(2+); but the toxic effect of effluents with higher levels of Hg(2+) was better detected by C. dubia. The most sensitive battery of bioassays to detect low concentrations of dissolved metals in effluents was the 72-h chronic test with C. vulgaris and the 48-h acute test with C. dubia. PMID:25199585

  19. Characterization of chemical waste site contamination and its extent using bioassays

    SciTech Connect

    Thomas, J.M.; Callahan, C.A.; Cline, J.F.; Greene, J.C.; McShane, M.C.; Miller, W.E.; Peterson, S.A.; Simpson, J.C.; Skalski, J.R.

    1984-12-01

    Bioassays were used in a three-phase research project to assess the comparative sensitivity of test organisms to known chemicals, determine if the chemical components in field soil and water samples containing unknown contaminants could be inferred from our laboratory studies using known chemicals, and to investigate kriging (a relatively new statistical mapping technique) and bioassays as methods to define the areal extent of chemical contamination. The algal assay generally was most sensitive to samples of pure chemicals, soil elutriates and water from eight sites with known chemical contamination. Bioassays of nine samples of unknown chemical composition from the Rocky Mountain Arsenal (RMA) site showed that a lettuce seed soil contact phytoassay was most sensitive. In general, our bioassays can be used to broadly identify toxic components of contaminated soil. Nearly pure compounds of insecticides and herbicides were less toxic in the sensitive bioassays than were the counterpart commercial formulations. This finding indicates that chemical analysis alone may fail to correctly rate the severity of environmental toxicity. Finally, we used the lettuce seed phytoassay and kriging techniques in a field study at RMA to demonstrate the feasibility of mapping contamination to aid in cleanup decisions. 25 references, 9 figures, 9 tables.

  20. Susceptibility of Bagrada hilaris (Hemiptera: Pentatomidae) to Insecticides in Laboratory and Greenhouse Bioassays.

    PubMed

    Palumbo, John C; Prabhaker, Nilima; Reed, Darcy A; Perring, Thomas M; Castle, Steven J; Huang, Ta-I

    2015-04-01

    Field-collected nymphs and adults of Bagrada hilaris (Burmeister) (Hemiptera: Penatatomidae) from three locations were evaluated for susceptibility to insecticides representing 10 classes of insecticide chemistry. Although relative susceptibilities differed between leaf-spray and leaf-dip Petri dish bioassays, consistently low LC50 values were determined for chlorpyrifos, bifenthrin, and lambda-cyhalothrin. Fenpropathrin and methomyl had intermediate values. Susceptibility to dinotefuran varied depending on the bioassay, possibly owing to leaf substrates used in the two bioassays. In soil systemic bioassays, the LC50 value of dinotefuran was significantly greater than that of two other neonicotinoids, imidacloprid and thiamethoxam, and the anthranilic diamide, cyantraniliprole. Mortality and feeding damage of B. hilaris and plant growth on insecticide-treated plants in greenhouse trials were consistent with the laboratory bioassays; the best results were seen with bifenthrin, methomyl, and chlorpyrifos. Mortality to the neonicotinoids was not evident; however, feeding damage and plant growth responses on dinotefuran-treated plants damage were similar to the noninfested control. This highlights the apparent antifeedant properties of dinotefuran that may have prevented adults from injuring broccoli plants after exposure to foliar spray residues. Data presented serve as baseline susceptibilities that can be used to monitor for resistance development in field populations of B. hilaris. PMID:26470178

  1. Studies on the bioassayable growth hormone-like activity of plasma

    NASA Technical Reports Server (NTRS)

    Ellis, S.; Vodian, M. A.; Grindeland, R. E.

    1978-01-01

    Evidence supporting the existence of bioassayable growth hormone-like activity in blood plasma distinct from the growth hormone measurable by radioimmunoassay and from somatomedin is presented. Tibial assays of the growth-hormone-like activity of injected, concentrated normal human and rat plasma in hypophysectomized rats reveal 200- and 50-fold activity excesses, respectively, with respect to the amount of growth hormone detected by radioimmunoassay. The origin of this bioassayable plasma hormone has been localized to the region of the pituitary, the origin of growth hormone, a distribution not followed by somatomedin C. Purification of the bioassayable agent indicates that is has a molecular weight of between 60,000 and 80,000, in contrast to that of growth hormone (20,000), and that the bioassayable activity is distinct from that of somatomedin C. Growth hormone-like activity detected in Cohn fraction IV as well as plasma activity, are found to be collectable on Dowex 50 resin, in contrast to somatomedin C and nonsuppressible insulin-like activity. The formation of bioassayable growth hormone-activity agents from radioimmunoassayable growth hormone and directly in the pituitary is suggested.

  2. Bioaccumulation of organic chemicals in contaminated soils: evaluation of bioassays with earthworms.

    PubMed

    Jager, Tjalling; van der Wal, Leon; Fleuren, Roel H L J; Barendregt, Arjan; Hermens, Joop L M

    2005-01-01

    Earthworms live in close contact with the soil and can thus be considered representative for the bioavailability of chemicals at contaminated sites. Bioavailability can either be assessed by analyzing earthworms from contaminated locations or by exposing laboratory-reared specimens to soil samples from the field (bioassays). In this study, we investigate the relevance of bioassays by using an extended experimental design (to identify signs of depletion of the bioavailable phase by the earthworms) and by using two species of earthworm (the standard test species Eisenia andrei and the field-relevant Aporrectodea caliginosa). Furthermore, bioassay results are compared to body residues of worms collected from the field site: a heavily polluted polder, amended with dredge spoil. We focused on telodrin, dieldrin, hexachlorobenzene, and eight PCBs. With our bioassay design, it was shown that depletion was unlikely, although more subtle effects could have occurred (e.g., changes in sorption during the experiments). E. andrei is a good choice for bioassays because its body residues correlate well to those in A. caliginosa, as well as to those in the field-collected worms. Nevertheless, E. andrei accumulated slightly more than the other species and appeared to be more sensitive to the conditions in soil from one of our sites. PMID:15667108

  3. Comparison of Citrated Human Blood, Citrated Sheep Blood, and Defibrinated Sheep Blood Mueller-Hinton Agar Preparations for Antimicrobial Susceptibility Testing of Streptococcus pneumoniae Isolates ▿

    PubMed Central

    Satzke, Catherine; Seduadua, Anna; Chandra, Reginald; Carapetis, Jonathan R.; Mulholland, E. Kim; Russell, Fiona M.

    2010-01-01

    The use of Mueller-Hinton agar supplemented with citrated human or citrated sheep blood was compared with the use of routinely used Mueller-Hinton agar supplemented with defibrinated sheep blood for antimicrobial susceptibility testing of Streptococcus pneumoniae. The alternate supplements were found to be unsatisfactory, particularly for testing resistant isolates, and therefore are not recommended. PMID:20668133

  4. Unusual non-fluorescent broad spectrum siderophore activity (SID EGYII) by Pseudomonas aeruginosa strain EGYII DSM 101801 and a new insight towards simple siderophore bioassay.

    PubMed

    Embaby, Amira M; Heshmat, Yasmin; Hussein, Ahmed

    2016-03-01

    Present study highlights an unusual non-fluorescent hydroxamate broad spectrum siderophore (SID EGYII) activity from Pseudomonas aeruginosa strain EGYII DSM 101801, a soil bacterial isolate, along with simple low cost effective siderophore bioassay. Detection of SID EGYII activity qualitatively was proved by masking this activity against Erwinia amylovora strain EGY1 DSM 101800, an indicator strain, in well-cut diffusion assay containing 100 µM FeCl3. SID EGYII activity was expressed quantitatively as arbitrary units [Siderophore arbitrary units (SAU)] 380 SAU/mL against E. amylovora strain EGY1 DSM 101800. Maximal SID EGYII activity was achieved upon growing P. aeruginosa strain EGYII DSM 101801 in PYB broth at 180 rpm for 24 h. SID EGYII displayed a broad spectrum antimicrobial activity against some human pathogens (i.e., Gram-positive bacteria, Gram-negative bacteria and yeasts) and a fireblight plant pathogen. Interestingly, transformants of Escherichia coli JM109 (DE3)pSID/EGYII harboring P. aeruginosa strain EGYII DSM 101801 plasmid demonstrated a perceivable antimicrobial activity against E. amylovora strain EGY1 DSM 101800. The broad spectrum antimicrobial activity of the unusual non-fluorescent SID EGYII would underpin its high potential in targeting bacterial pathogens posing probable threats to human health and agricultural economy. The present simple low cost effective bioassay is a new insight towards an alternative to the expensive cumbersome siderophore Chrome Azurol S assay. PMID:27015845

  5. Quantum diffusion

    SciTech Connect

    Habib, S.

    1994-10-01

    We consider a simple quantum system subjected to a classical random force. Under certain conditions it is shown that the noise-averaged Wigner function of the system follows an integro-differential stochastic Liouville equation. In the simple case of polynomial noise-couplings this equation reduces to a generalized Fokker-Planck form. With nonlinear noise injection new ``quantum diffusion`` terms rise that have no counterpart in the classical case. Two special examples that are not of a Fokker-Planck form are discussed: the first with a localized noise source and the other with a spatially modulated noise source.

  6. DIFFUSION PUMP

    DOEpatents

    Levenson, L.

    1963-09-01

    A high-vacuum diffusion pump is described, featuring a novel housing geometry for enhancing pumping speed. An upright, cylindrical lower housing portion is surmounted by a concentric, upright, cylindrical upper housing portion of substantially larger diameter; an uppermost nozzle, disposed concentrically within the upper portion, is adapted to eject downwardly a conical sheet of liquid outwardly to impinge upon the uppermost extremity of the interior wall of the lower portion. Preferably this nozzle is mounted upon a pedestal rising coaxially from within the lower portion and projecting up into said upper portion. (AEC)

  7. Preparation and application of agar/alginate/collagen ternary blend functional food packaging films.

    PubMed

    Wang, Long-Feng; Rhim, Jong-Whan

    2015-09-01

    Ternary blend agar/alginate/collagen (A/A/C) hydrogel films with silver nanoparticles (AgNPs) and grapefruit seed extract (GSE) were prepared. Their performance properties, transparency, tensile strength (TS), water vapor permeability (WVP), water contact angle (CA), water swelling ratio (SR), water solubility (WS), and antimicrobial activity were determined. The A/A/C film was highly transparent, and both AgNPs and GSE incorporated blend films (A/A/C(AgNPs) and A/A/C(GSE)) exhibited UV-screening effect, especially, the A/A/C(GSE) film had high UV-screening effect without sacrificing the transmittance. In addition, the A/A/C blend films formed efficient hydrogel film with the water holding capacity of 23.6 times of their weight. Both A/A/C(AgNPs) and A/A/C(GSE) composite films exhibited strong antimicrobial activity against both Gram-positive (Listeria monocytogenes) and Gram-negative (Escherichia coli) food-borne pathogenic bacteria. The test results of fresh potatoes packaging revealed that all the A/A/C ternary blend films prevented forming of condensed water on the packaged film surface, both A/A/C(AgNPs) and A/A/C(GSE) composite films prevented greening of potatoes during storage. The results indicate that the ternary blend hydrogel films incorporated with AgNPs or GSE can be used not only as antifogging packaging films for highly respiring fresh agriculture produce, but also as an active food packaging system utilizing their strong antimicrobial activity. PMID:26187189

  8. Comparative performance of Thin Layer Agar and Löwenstein-Jensen culture for diagnosis of tuberculosis.

    PubMed

    Battaglioli, T; Rintiswati, N; Martin, A; Palupi, K R; Bernaerts, G; Dwihardiani, B; Ahmad, R A; Matthys, F; Mahendradhata, Y; Van der Stuyft, P

    2013-11-01

    Sputum smear microscopy for the diagnosis of tuberculosis (TB) is cheap and simple but its sensitivity is low. Culture on Löwenstein-Jensen (LJ) is more sensitive but it takes a long time to yield results. Thin-Layer Agar (TLA) culture was suggested as an equally sensitive and faster alternative. We evaluated the performance of TLA for diagnosing TB in Jogjakarta, Indonesia. People with suspected TB presenting from July 2010 to July 2011 to two chest clinics of the National TB Control Programme network of Jogjakarta were eligible for inclusion. A sputum sample was sent to the Gadjah Mada University microbiology laboratory for concentration, smearing, Ziehl-Neelsen staining and culture on LJ and TLA. Sensitivity of cultures was evaluated against a composite reference standard (any positive culture). Time to detection of Mycobacteria was recorded. Out of 1414 samples, 164 (12%) were smear positive, 99 (7%) were scanty and 1151 (81%) were negative. On TLA and LJ respectively, 168 (12%) and 149 (11%) samples were positive, 72 (5%) and 32 (2%) were contaminated (κ = 0.64; 95% CI 0.59-0.69, p <0.01). Using the reference standard, 196 (14%) TB cases were identified. The sensitivity of TLA was 0.86 (95% CI 0.80-0.90), significantly higher (p 0.03) than for LJ (0.76; 95% CI 0.69-0.81). The median time to detection in days was significantly shorter (p <0.01) for TLA (12; 95% CI 11-13) than for LJ (44; 95% CI 43-45). TLA is a rapid and sensitive method for the diagnosis of TB. Implementation studies to evaluate the cost-effectiveness and impact of its introduction into programmatic settings are urgently needed. PMID:23738759

  9. Strategies for Transferring Mixtures of Organic Contaminants from Aquatic Environments into Bioassays.

    PubMed

    Jahnke, Annika; Mayer, Philipp; Schäfer, Sabine; Witt, Gesine; Haase, Nora; Escher, Beate I

    2016-06-01

    Mixtures of organic contaminants are ubiquitous in the environment. Depending on their persistence and physicochemical properties, individual chemicals that make up the mixture partition and distribute within the environment and might then jointly elicit toxicological effects. For the assessment and monitoring of such mixtures, a variety of cell-based in vitro and low-complexity in vivo bioassays based on algae, daphnids or fish embryos are available. A very important and sometimes unrecognized challenge is how to combine sampling, extraction and dosing to transfer the mixtures from the environment into bioassays, while conserving (or re-establishing) their chemical composition at adjustable levels for concentration-effect assessment. This article outlines various strategies for quantifiable transfer from environmental samples including water, sediment, and biota into bioassays using total extraction or polymer-based passive sampling combined with either solvent spiking or passive dosing. PMID:26804122

  10. Bioassay, isolation and studies on the mechanism of action of neurite extension factor

    NASA Technical Reports Server (NTRS)

    Kligman, D.

    1984-01-01

    The identification and purification of molecules active in promoting neurite outgrowth requires a sensitive reproducible bioassay. A quantitative bioassay was utilized to purify a neurite extension factor (NEF) based on counting the number of phase bright neurons with processes at least equal to one cell body diameter after 20 hrs. in culture is defined, serum free medium. Using a combination of heat treatment DEAE cellulose chromatography and gel filtration, an acidic protein of M sub r = 75,000 was highly purified. Upon reduction, it yields subunits of M sub r = 37,000. Purified fractions are active half maximally at 100 ng/ml in inducing neurite outgrowth in this bioassay. Currently, monoclonal antibodies to NEF are being produced. Female Balb C mice were immunized with the antigen and fusions with mouse myeloma cells will be performed to yield hybridoma cells.

  11. Comparison of liquid chromatographic and bioassay procedures for determining depletion of intramuscularly injected tylosin.

    PubMed

    Moats, W A; Harris, E W; Steele, N C

    1985-01-01

    Crossbred pigs weighing 80-110 kg were injected intramuscularly in the ham with 8.8 mg/kg tylosin. Animals were slaughtered in groups of 3 at intervals of 4 h, and 1, 2, 4, and 8 days after injection, and samples of blood, injected muscle, uninjected muscle, liver, and kidney were analyzed by liquid chromatography (LC) and by bioassay using Sarcina lutea as the test organism. The LC method was far more sensitive with a detection limit of less than 0.1 ppm, while the detection limit by bioassay was about 0.5 ppm in tissue. Results by bioassay and LC sometimes differed considerably for tissue samples. Residues in all tissues were below the tolerance limit of 0.2 ppm at 24 h, except in the injected muscle in one animal. Residues were not detected in any tissue of any animal at 48 h after treatment. PMID:4019360

  12. Bioassay for estimating the biogenic methane-generating potential of coal samples

    USGS Publications Warehouse

    Jones, E.J.P.; Voytek, M.A.; Warwick, P.D.; Corum, M.D.; Cohn, A.; Bunnell, J.E.; Clark, A.C.; Orem, W.H.

    2008-01-01

    Generation of secondary biogenic methane in coal beds is likely controlled by a combination of factors such as the bioavailability of coal carbon, the presence of a microbial community to convert coal carbon to methane, and an environment supporting microbial growth and methanogenesis. A set of treatments and controls was developed to bioassay the bioavailability of coal for conversion to methane under defined laboratory conditions. Treatments included adding a well-characterized consortium of bacteria and methanogens (enriched from modern wetland sediments) and providing conditions to support endemic microbial activity. The contribution of desorbed methane in the bioassays was determined in treatments with bromoethane sulfonic acid, an inhibitor of microbial methanogenesis. The bioassay compared 16 subbituminous coal samples collected from beds in Texas (TX), Wyoming (WY), and Alaska (AK), and two bituminous coal samples from Pennsylvania (PA). New biogenic methane was observed in several samples of subbituminous coal with the microbial consortium added, but endemic activity was less commonly observed. The highest methane generation [80????mol methane/g coal (56??scf/ton or 1.75??cm3/g)] was from a south TX coal sample that was collected from a non-gas-producing well. Subbituminous coals from the Powder River Basin, WY and North Slope Borough, AK contained more sorbed (original) methane than the TX coal sample and generated 0-23????mol/g (up to 16??scf/ton or 0.5??cm3/g) new biogenic methane in the bioassay. Standard indicators of thermal maturity such as burial depth, nitrogen content, and calorific value did not explain differences in biogenic methane among subbituminous coal samples. No original methane was observed in two bituminous samples from PA, nor was any new methane generated in bioassays of these samples. The bioassay offers a new tool for assessing the potential of coal for biogenic methane generation, and provides a platform for studying the

  13. In vitro bioassays to evaluate complex chemical mixtures in recycled water

    PubMed Central

    Jia, Ai; Escher, Beate I.; Leusch, Frederic D.L.; Tang, Janet Y.M.; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M.; Snyder, Shane A.

    2016-01-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, aryl hydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  14. Genotoxicity of soil from farmland irrigated with wastewater using three plant bioassays.

    PubMed

    Cabrera, G L; Rodriguez, D M

    1999-05-19

    Three well known plant bioassays, the Allium root chromosome aberration (AL-RAA) assay, the Tradescantia micronucleus (Trad-MCN) assay, and the Tradescantia stamen hair (Trad-SHM) mutation assay were validated in 1991 by the International Programme on Chemical Safety (IPCS) under the auspices of the World Health Organization, and the United Nations Environment Programme (UNEP). These plant bioassays have proven to be efficient tests for chemical screening and especially for in situ monitoring for genotoxicity of environmental pollutants. As a result of this validation study, standard protocols of these three plant bioassays were used by some of the 11 participating countries in the IPCS to carry on genotoxicity tests on air, water and soil as a follow up activity. In the city of Queretaro, Mexico, wastewater coming from both industrial and domestic sources and without any treatment is used to irrigate the farm crops, polluting the soil. Potentially the pollutants could reach the food chain. For the above reason, soil irrigated with wastewater was sampled and monitored for the presence of genotoxic agents using the above three bioassays. Extracts from soil samples were made using distilled water and organic solvents by shaking the sample for about 12 h under a relatively low temperature (15-20 degrees C). Plant cuttings of Tradescantia or the roots of Allium were treated by submerging them in the extracts. Three replicates of each sample were analyzed in each of the three bioassays. Extracts using DMSO, ethanol and distilled water tested positive in the three bioassays and there were no differences for the genotoxicity of the extracts with the different solvents. PMID:10350600

  15. In vitro bioassays to evaluate complex chemical mixtures in recycled water.

    PubMed

    Jia, Ai; Escher, Beate I; Leusch, Frederic D L; Tang, Janet Y M; Prochazka, Erik; Dong, Bingfeng; Snyder, Erin M; Snyder, Shane A

    2015-09-01

    With burgeoning population and diminishing availability of freshwater resources, the world continues to expand the use of alternative water resources for drinking, and the quality of these sources has been a great concern for the public as well as public health professionals. In vitro bioassays are increasingly being used to enable rapid, relatively inexpensive toxicity screening that can be used in conjunction with analytical chemistry data to evaluate water quality and the effectiveness of water treatment. In this study, a comprehensive bioassay battery consisting of 36 bioassays covering 18 biological endpoints was applied to screen the bioactivity of waters of varying qualities with parallel treatments. Samples include wastewater effluent, ultraviolet light (UV) and/or ozone advanced oxidation processed (AOP) recycled water, and infiltrated recycled groundwater. Based on assay sensitivity and detection frequency in the samples, several endpoints were highlighted in the battery, including assays for genotoxicity, mutagenicity, estrogenic activity, glucocorticoid activity, arylhydrocarbon receptor activity, oxidative stress response, and cytotoxicity. Attenuation of bioactivity was found to be dependent on the treatment process and bioassay endpoint. For instance, ozone technology significantly removed oxidative stress activity, while UV based technologies were most efficient for the attenuation of glucocorticoid activity. Chlorination partially attenuated genotoxicity and greatly decreased herbicidal activity, while groundwater infiltration efficiently attenuated most of the evaluated bioactivity with the exception of genotoxicity. In some cases, bioactivity (e.g., mutagenicity, genotoxicity, and arylhydrocarbon receptor) increased following water treatment, indicating that transformation products of water treatment may be a concern. Furthermore, several types of bioassays with the same endpoint were compared in this study, which could help guide the selection

  16. False-Positive Serum Botulism Bioassay in Miller-Fisher Syndrome.

    PubMed

    Zeylikman, Yuriy; Shah, Vishal; Shah, Umang; Mirsen, Thomas R; Campellone, Joseph V

    2015-09-01

    We describe a patient with acute progressive weakness and areflexia. Both botulism and Miller-Fisher variant of Guillain-Barré syndrome were initial diagnostic considerations, and she was treated with intravenous immunoglobulin and botulinum antitoxin. A mouse bioassay was positive for botulinum toxin A, although her clinical course, electrodiagnostic studies, and cerebrospinal fluid findings supported Miller-Fisher syndrome. This patient's atypical features offer points of discussion regarding the evaluation of patients with acute neuromuscular weakness and emphasize the limitations of the botulism bioassay. PMID:26301377

  17. A field bioassay to evaluate potential spatial repellents against natural mosquito populations.

    PubMed

    Chauhan, K R; Aldrich, J R; McCardle, P W; White, G B; Webb, R E

    2012-12-01

    A field bioassay evaluating candidate chemicals as aerial repellents was developed and evaluated against natural mosquito populations in Beltsville, MD. The bioassay consisted of an attractive source surrounded by a grid of 16 septa containing a volatile candidate aerial repellent, compared with an attractive source without such a grid. The attractive source was a Centers for Disease Control and Prevention light trap supplemented with carbon dioxide. Significant sources of variation included weather, position, and the differential response of mosquito species. Despite these sources of variation, significant repellent responses were obtained for catnip oil, E,Z-dihydronepetalactone, and DEET. PMID:23393752

  18. A Brine Shrimp Bioassay for Measuring Toxicity and Remediation of Chemicals

    NASA Astrophysics Data System (ADS)

    Lieberman, Marya

    1999-12-01

    A bioassay using Artemia franciscana (brine shrimp) was adapted to measure the toxicity of household chemicals. One project is described in which students collect dose-response curves for seven commercial flea-killing products. Next, groups of students researched the insecticidal ingredients of the flea products. On the basis of the structures of the active ingredients, they chose remediation methods to make the flea product less toxic to brine shrimp; procedures included copper-catalyzed hydrolysis, adsorption onto activated charcoal, bleach treatment, and photodegradation. No special equipment or supplies are necessary for the bioassay other than the brine shrimp eggs, which can be obtained at any aquarium store.

  19. A versatile electrowetting-based digital microfluidic platform for quantitative homogeneous and heterogeneous bio-assays

    NASA Astrophysics Data System (ADS)

    Vergauwe, Nicolas; Witters, Daan; Ceyssens, Frederik; Vermeir, Steven; Verbruggen, Bert; Puers, Robert; Lammertyn, Jeroen

    2011-05-01

    Electrowetting-on-dielectric (EWOD) lab-on-a-chip systems have already proven their potential within a broad range of bio-assays. Nevertheless, research on the analytical performance of those systems is limited, yet crucial for a further breakthrough in the diagnostic field. Therefore, this paper presents the intrinsic possibilities of an EWOD lab-on-a-chip as a versatile platform for homogeneous and heterogeneous bio-assays with high analytical performance. Both droplet dispensing and splitting cause variations in droplet size, thereby directly influencing the assay's performance. The extent to which they influence the performance is assessed by a theoretical sensitivity analysis, which allows the definition of a basic framework for the reduction of droplet size variability. Taking advantage of the optimized droplet manipulations, both homogeneous and heterogeneous bio-assays are implemented in the EWOD lab-on-a-chip to demonstrate the analytical capabilities and versatility of the device. A fully on-chip enzymatic assay is realized with high analytical performance. It demonstrates the promising capabilities of an EWOD lab-on-a-chip in food-related and medical applications, such as nutritional and blood analyses. Further, a magnetic bio-assay for IgE detection using superparamagnetic nanoparticles is presented whereby the nanoparticles are used as solid carriers during the bio-assay. Crucial elements are the precise manipulation of the superparamagnetic nanoparticles with respect to dispensing and separation. Although the principle of using nano-carriers is demonstrated for protein detection, it can be easily extended to a broader range of bio-related applications like DNA sensing. In heterogeneous bio-assays the chip surface is actively involved during the execution of the bio-assay. Through immobilization of specific biological compounds like DNA, proteins and cells a reactive chip surface is realized, which enhances the bio-assay performance. To demonstrate

  20. Review of Bioassays for Monitoring Fate and Transport ofEstrogenic Endocrine Disrupting Compounds in Water

    SciTech Connect

    CGCampbell@lbl.gov

    2004-01-30

    Endocrine disrupting compounds (EDCs) are recognizedcontaminants threatening water quality. Despite efforts in sourceidentification, few strategies exist for characterization or treatment ofthis environmental pollution. Given that there are numerous EDCs that cannegatively affect humans and wildlife, general screening techniques likebioassays and biosensors provide an essential rapid and intensiveanalysis capacity. Commonly applied bioassays include the ELISA and YESassays, but promising technologies include ER-CALUXa, ELRA, Endotecta,RIANA, and IR-bioamplification. Two biosensors, Endotecta and RIANA, arefield portable using non-cellular biological detection strategies.Environmental management of EDCs in water requires integration ofbiosensors and bioassays for monitoring and assessment.

  1. [Tryptose sulphite cycloserine agar for the recovery of Clostridium perfringens in surface waters: a study of different modes of utilization].

    PubMed

    Nusca, A; Orefice, L; Paradiso, R

    2007-01-01

    In the recent European Drinking Water Directive, Clostridium perfringens has assumed increasing importance so as to be considered a primary contamination indicator. Therefore it emerged the necessity to make culture methods, aimed at its recovery, more specific and sensitive. In this study we have verified the ability of Tryptose Sulphite Cycloserine Agar plates (TSC Agar), prepared and stored before the use at refrigeration temperature (+4 degrees) for different times, to show typical colonies, using both, the single layer and double layer techniques. Results show that storage of the prepared medium, even for a few days, decrease the recovery of typical colonies although such negative effect is minimized by using the double layer technique. PMID:17405507

  2. Preparation of agar nanospheres: comparison of response surface and artificial neural network modeling by a genetic algorithm approach.

    PubMed

    Zaki, Mohammad Reza; Varshosaz, Jaleh; Fathi, Milad

    2015-05-20

    Multivariate nature of drug loaded nanospheres manufacturing in term of multiplicity of involved factors makes it a time consuming and expensive process. In this study genetic algorithm (GA) and artificial neural network (ANN), two tools inspired by natural process, were employed to optimize and simulate the manufacturing process of agar nanospheres. The efficiency of GA was evaluated against the response surface methodology (RSM). The studied responses included particle size, poly dispersity index, zeta potential, drug loading and release efficiency. GA predicted greater extremum values for response factors compared to RSM. However, real values showed some deviations from predicted data. Appropriate agreement was found between ANN model predicted and real values for all five response factors with high correlation coefficients. GA was more successful than RSM in optimization and along with ANN were efficient tools in optimizing and modeling the fabrication process of drug loaded in agar nanospheres. PMID:25817674

  3. Effect of EDTA on Pb(II) Uptake and Translocation by Tumbleweed (Salsola Kali): Agar and Hydroponics Studies

    SciTech Connect

    de la Rosa, Guadalupe; Gardea-Torresdey, Jorge L.; Peralta-Videa, Jose R.; Aldrich, Mary

    2004-03-31

    Environmental accumulation of Pb represents a worldwide health hazard. While conventional cleanup techniques are generally expensive and soil disturbing, phytoremediation represents an inexpensive friendly option for the removal of contaminants from soil and water. In this research, tumbleweed (Salsola kali) plants exposed for 15 days to Pb(NO3)2 at 80 and 125 ppm in hydroponics and agar media, demonstrated a high capacity to uptake lead. The results showed that the plants cultivated in agar accumulated 25563, 5534 and 2185 mg Pb kg-1 DW in roots, stems and leaves, respectively. Moreover, Pb concentrations found in hydroponically grown tumbleweed plants tissues were 30744, 1511 and 1421 mg kg-1 DW in roots, stems and leaves, respectively. It was observed that EDTA enhanced Pb translocation. No Pb phytotoxic effects were observed during the experimental time period. Cellular structural features were also observed using TEM.

  4. [Clinical symptoms and circumastances of acute poisonings with fly agaric (Amanita muscaria) and panther cap (Amanita pantherina)].

    PubMed

    Łukasik-Głebocka, Magdalena; Druzdz, Artur; Naskret, Maciej

    2011-01-01

    Mushroom poisonings in Poland are quite common, especially in summer and autumn, but fly agaric (Amanita muscaria) and panther cap (Amanita pantherina) are rather rare cause of these intoxications. Fly agaric is a cause of deliberate poisoning, whereas panther cap poisoning also happens accidentally. The main toxins of these two mushrooms are ibotenic acid (pantherine, agarine), muscimol, muscazone and muscaridine. The other bioactive substances are stizolobic and stizolobinic acids and aminodicarboxyethylthiopropanoic acids. All these compounds are responsible for diverse picture of intoxication. An analysis of patients with Amanita muscaria and Amanita pantherina poisoning hospitalized in the Poznan Departament of Toxicology revealed that symptoms occurred after 30 minutes to 2 hours with vomiting, hallucinations, restlessness, increased psychomotor drive and central nervous system depression. Other antycholinergic symptoms like tachycardia and increased blood pressure, mydriasis, dry and red skin were seen only in a few cases. Acute respiratory failure was the most dangerous symptom observed in the course of poisoning. PMID:22010435

  5. Use of hydrogen peroxide treatment and crystal violet agar plates for selective recovery of bacteriophages from natural environments

    SciTech Connect

    Asghari, A.; Farrah, S.R.; Bitton, G. )

    1992-04-01

    Hydrogen peroxide inactivated bacteriophages and bacteria at different rates. A concentration of 0.1% hydrogen peroxide reduced the numbers of several bacteria by an average of 94% but caused an average of 25% inactivation in the numbers of bacteriophages tested. Treating natural samples with hydrogen peroxide selectively reduced the indigenous bacterial flora and permitted better visualization of plaques of lawns of Escherichia coli C-3000. In some cases indigenous gram-positive bacteria were relatively resistant to hydrogen peroxide, but their growth could be limited by incorporation of crystal violet into the bottom agar used for plaque assays. The use of hydrogen peroxide treatment and crystal violet-containing plates permitted recovery of more phages from natural samples than did other procedures, such as chloroform pretreatment or the use of selective plating agar such as EC medium.

  6. Disk Diffusion Testing Using Candida sp. Colonies Taken Directly from CHROMagar Candida Medium May Decrease Time Required To Obtain Results

    PubMed Central

    Klevay, Michael; Ebinger, Alex; Diekema, Daniel; Messer, Shawn; Hollis, Richard; Pfaller, Michael

    2005-01-01

    We compared results of disk diffusion antifungal susceptibility testing from Candida sp. strains passaged on CHROMagar and on potato dextrose agar. The overall categorical agreements for fluconazole and voriconazole disk testing were 95% and 98% with 0% and 0.5% very major errors, respectively. Disk diffusion testing by the CLSI (formerly NCCLS) M44-A method can be performed accurately by taking inocula directly from CHROMagar. PMID:16000489

  7. Anti-bacterial activity and brine shrimp lethality bioassay of methanolic extracts of fourteen different edible vegetables from Bangladesh

    PubMed Central

    Ullah, M. Obayed; Haque, Mahmuda; Urmi, Kaniz Fatima; Zulfiker, Abu Hasanat Md.; Anita, Elichea Synthi; Begum, Momtaj; Hamid, Kaiser

    2013-01-01

    Objective To investigate the antibacterial and cytotoxic activity of fourteen different edible vegetables methanolic extract from Bangladesh. Methods The antibacterial activity was evaluated using disc diffusion assay method against 12 bacteria (both gram positive and gram negative). The plant extracts were also screened for cytotoxic activity using the brine shrimp lethality bioassay method and the lethal concentrations (LC50) were determined at 95% confidence intervals by analyzing the data on a computer loaded with “Finney Programme”. Results All the vegetable extracts showed low to elevated levels of antibacterial activity against most of the tested strains (zone of inhibition=5-28 mm). The most active extract against all bacterial strains was from Xanthium indicum which showed remarkable antibacterial activity having the diameter of growth inhibition zone ranging from 12 to 28 mm followed by Alternanthera sessilis (zone of inhibition=6-21 mm). All extracts exhibited considerable general toxicity towards brine shrimps. The LC50 value of the tested extracts was within the range of 8.447 to 60.323 µg/mL with respect to the positive control (vincristine sulphate) which was 0.91 µg/mL. Among all studied extracts, Xanthium indicum displayed the highest cytotoxic effect with LC50 value of 8.447 µg/mL. Conclusions The results of the present investigation suggest that most of the studied plants are potentially good source of antibacterial and anticancer agents. PMID:23570009

  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. 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

  10. Liofilchem® O.A. Listeria agar and direct CAMP test provided sooner Listeria monocytogenes identification from neonatal bacteremia

    PubMed Central

    Savini, Vincenzo; Marrollo, Roberta; Serio, Annalisa; Paparella, Antonello; Argentieri, Angela Valentina; D’Antonio, Marianna; Coclite, Eleonora; Fusilli, Paola; Fazii, Paolo

    2014-01-01

    Listeria monocytogenes infection in pregnant women and newborns is a cause for serious concern, and invasive disease outcome strongly depends on prompt antibiotic therapy. To provide sooner identification from neonatal bacteremia we performed a CAMP test directly on positive blood aliquots and inoculated the Liofilchem® O.A. Listeria chromogenic agar as well, thus providing a 24-h turn-around time for response. PMID:24695762

  11. Nutrient limitation leads to penetrative growth into agar and affects aroma formation in Pichia fabianii, P. kudriavzevii and Saccharomyces cerevisiae.

    PubMed

    van Rijswijck, Irma M H; Dijksterhuis, Jan; Wolkers-Rooijackers, Judith C M; Abee, Tjakko; Smid, Eddy J

    2015-01-01

    Among fermentative yeast species, Saccharomyces cerevisiae is most frequently used as a model organism, although other yeast species may have special features that make them interesting candidates to apply in food-fermentation processes. In this study, we used three yeast species isolated from fermented masau (Ziziphus mauritiana) fruit, S. cerevisiae 131, Pichia fabianii 65 and Pichia kudriavzevii 129, and determined the impact of nitrogen and/or glucose limitation on surface growth mode and the production of volatile organic compounds (VOCs). All three species displayed significant changes in growth mode in all nutrient-limited conditions, signified by the formation of metafilaments or pseudohyphae. The timing of the transition was found to be species-specific. Transition in growth mode is suggested to be linked to the production of certain fusel alcohols, such as phenylethyl alcohol, which serve as quorum-sensing molecules. Interestingly, we did not observe concomitant increased production of phenylethyl alcohol and filamentous growth. Notably, a broader range of esters was found only for the Pichia spp. grown on nitrogen-limited agar for 21 days compared to nutrient-rich agar, and when grown on glucose- and glucose- plus nitrogen-limited agar. Our data suggest that for the Pichia spp., the formation of esters may play an important role in the switch in growth mode upon nitrogen limitation. Further biological or ecological implications of ester formation are discussed. PMID:25308873

  12. Beyond Agar: Gel Substrates with Improved Optical Clarity and Drug Efficiency and Reduced Autofluorescence for Microbial Growth Experiments

    PubMed Central

    Jaeger, Philipp A.; McElfresh, Cameron; Wong, Lily R.

    2015-01-01

    Agar, a seaweed extract, has been the standard support matrix for microbial experiments for over a century. Recent developments in high-throughput genetic screens have created a need to reevaluate the suitability of agar for use as colony support, as modern robotic printing systems now routinely spot thousands of colonies within the area of a single microtiter plate. Identifying optimal biophysical, biochemical, and biological properties of the gel support matrix in these extreme experimental conditions is instrumental to achieving the best possible reproducibility and sensitivity. Here we systematically evaluate a range of gelling agents by using the yeast Saccharomyces cerevisiae as a model microbe. We find that carrageenan and Phytagel have superior optical clarity and reduced autofluorescence, crucial for high-resolution imaging and fluorescent reporter screens. Nutrient choice and use of refined Noble agar or pure agarose reduce the effective dose of numerous selective drugs by >50%, potentially enabling large cost savings in genetic screens. Using thousands of mutant yeast strains to compare colony growth between substrates, we found no evidence of significant growth or nutrient biases between gel substrates, indicating that researchers could freely pick and choose the optimal gel for their respective application and experimental condition. PMID:26070672

  13. A soft agar colony assay for Lewis lung tumour and B16 melanoma taken directly from the mouse.

    PubMed Central

    Courtenay, V. D.

    1976-01-01

    A soft agar colony assay has been developed for the B16 mouse melanoma and the Lewis lung tumour. The special features of the technique are the use of a gas phase with 5% O2 instead of air and the addition of rat red blood cells. Single cell suspensions are prepared by trypsinization from the solid tumour and the cells are plated out in 0-3% agar over a layer of 0-5% agar in 30-mm Petri dishes. After 8 to 15 days' incubation in 5% O2, colonies of more than 50 cells are produced. Plating efficiencies of between 30 and 50% are usually obtained. The addition of up to 10(4) heavily irradiated tumour cells gives some further improvement in plating efficiency for the B16 melanoma but not for the Lewis lung tumour. Applications of the technique to measure cell survival in the two tumours after treatment with cytotoxic drugs and radiation are reported. The scatter of experimental points is relatively small, and in comparative experiments good agreement has been obtained with results using in vivo assay techniques. PMID:782495

  14. 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. PMID:9153733

  15. Irradiation of silver and agar/silver nanoparticles with argon, oxygen glow discharge plasma, and mercury lamp.

    PubMed

    Ahmad, Mahmoud M; Abdel-Wahab, Essam A; El-Maaref, A A; Rawway, Mohammed; Shaaban, Essam R

    2014-01-01

    The irradiation effect of argon, oxygen glow discharge plasma, and mercury lamp on silver and agar/silver nanoparticle samples is studied. The irradiation time dependence of the synthesized silver and agar/silver nanoparticle absorption spectra and their antibacterial effect are studied and compared. In the agar/silver nanoparticle sample, as the irradiation time of argon glow discharge plasma or mercury lamp increases, the peak intensity and the full width at half maximum, FWHM, of the surface plasmon resonance absorption band is increased, however a decrease of the peak intensity with oxygen glow plasma has been observed. In the silver nanoparticle sample, as the irradiation time of argon, oxygen glow discharge plasma or mercury lamp increases, the peak intensity of the surface plasmon resonance absorption band is increased, however, there is no significant change in the FWHM of the surface plasmon resonance absorption band. The SEM results for both samples showed nanoparticle formation with mean size about 50 nm and 40 nm respectively. Throughout the irradiation time with the argon, oxygen glow discharge plasma or mercury lamp, the antibacterial activity of several kinds of Gram-positive and Gram-negative bacteria has been examined. PMID:25184109

  16. Laboratory bioassay for assessing the effects of sludge supernatant on plant growth and vesicular-arbuscular mycorrhiza formation

    SciTech Connect

    Bohn, K.S.; Liberta, A.E.

    1982-12-01

    A laboratory bioassay is described for assessing the effects of sludge supernatant on juvenile corn growth and the ability of vesicular-arbuscular (VA) mycorrhizal fungi, indigenous to coal spoil, to form mycorrhizae. The bioassay demonstrated that application rates can be identified that have the potential to promote increased plant dry weight without suppressing the formation of VA mycorrhizae in a plant's root system.

  17. INITIATION/PROMOTION BIOASSAY IN RAT LIVER: USE OF GAMMA GLUTAMYLTRANSPEPTIDASE-POSITIVE FOCI TO INDICATE CARCINOGENIC ACTIVITY

    EPA Science Inventory

    Gamma Glutamyltranspeptidase (GGTase)-positive foci have been used to indicate activity in an initiation/promotion bioassay in rat liver. This rat liver foci bioassay has been proposed for inclusion in tier 2 of a three tier decision tree approach to carcinogenesis testing where ...

  18. Minimization of Between-well Sample Variance of Antifungal Activity Measurements Using a High-Throughput Screening Microplate Bioassay

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The use of microplate bioassays, or broth microdilution assays, to measure the activity of biological and synthetic compounds against fungal pathogens has increased in recent years; this technique has been identified as the most promising in vitro bioassay for quantifying antifungal activity. Quant...

  19. Evaluating macroinvertebrate population and community level effects in outdoor microcosms: Use of in situ bioassays and multivariate analysis

    SciTech Connect

    Shaw, J.L.; Manning, J.P.

    1996-05-01

    Evaluating toxicant effects on aquatic communities is difficult due to the ecological complexity at higher levels of organization. Two methods were assessed to improve the understanding of effects on macroinvertebrate communities in aquatic model ecosystems. First, in situ bioassay population effects were used to interpret effects at a higher organization level. Second, canonical discriminant analysis was used to investigate effects on community structure. In situ bioassays were conducted on six occasions in 17-m{sup 3} microcosms treated with copper sulfate. Macroinvertebrates occurring naturally in the microcosms were monitored. Epibenthic in situ bioassays were conducted using Caenis sp. (Ephemeroptera) and Hyalella azteca (Amphipoda) and a water column bioassay was conducted using Notonectidae (Hemiptera). Survival and growth were assessed after 3 d. Effects of copper on both notonectidae and Caenis were observed following application. However, the final Caenis epibenthic bioassays indicated that potential for recovery and survival was {ge}95%. Potential for recovery was less distinct in the water column bioassays. Copper effects also occurred on epibenthic macroinvertebrate populations and communities. Only four taxa, including Caenis, distinguished community differences among copper treatments soon after application. Later, communities showed similarities to the pretreatment bioassay. However, actual recovery was less apparent than the potential for recovery indicated by the bioassays, and community differences due to Caenis persisted.

  20. COMPARISON OF BIOASSAY AND ENZYME-LINKED IMMUNOSORBENT ASSAY FOR QUANTIFICATION OF 'SPODOPTERA FRUGIPERDA' NUCLEAR POLYHEDROSIS VIRUS IN SOIL

    EPA Science Inventory

    Standard curves with known amounts of Spodoptera frugiperda nuclear polyhedrosis virus (NPV) in soil were established with a bioassay and with an enzyme-linked immunosorbent assay (ELISA). The bioassay detected as few as 4 x 10 to the 4th power polyhedral inclusion bodies (PIB)/g...