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1.
The current AOAC use-dilution methods of disinfectant efficacy testing require the use of 48-54 h unadjusted broth cultures of Salmonella choleraesuis, Staphylococcus aureus, and Pseudomonas aeruginosa for the inoculation of stainless steel penicylinders. The use of unadjusted broth cultures contributes to noncomparable numbers of organisms on penicylinders among the test strains due to relative efficacy of bacterial attachment to penicylinders and to bacterial numbers in broth. To achieve comparable numbers of cells on the penicylinders among the 3 test strains, the cell densities of S. aureus and P. aeruginosa in broth culture were visually adjusted. Growth studies were conducted using S. choleraesuis and P. aeruginosa to determine the numbers of cells in broth at timed intervals and the corresponding numbers of cells attaching to the penicylinders. Results showed that the use of the 24 h broth cultures for all 3 test strains, with adjustment of S. aureus and P. aeruginosa broths, contributes to more comparable numbers of organisms attached to the penicylinders used in disinfectant testing.  相似文献   

2.
Two possible deficiencies in the AOAC use-dilution method for registration of chemical disinfectants by the Environmental Protection Agency are examined: (1) the physical disparities among brands of penicylinders and (2) the variability of bacterial numbers on penicylinders depending upon test strain and penicylinder surface texture. Textural differences of 2 brands of stainless steel penicylinders, one brand of porcelain, and one brand of glass were assessed by scanning electron microscopy. A considerable variation in smoothness of both inner and outer surfaces of stainless steel and porcelain penicylinders was observed. Glass penicylinders were very smooth. Numbers of bacteria attached to a penicylinder were assessed by vortexing the penicylinders 30 s at No. 4 after using the AOAC method of bacterial inoculation and drying 40 min at 37 degrees C. With this methodology, stainless steel carriers retained the 3 AOAC-recommended bacterial test strains differentially: ca 10(7) for Pseudomonas aeruginosa, 5 X 10(6) for Staphylococcus aureus, and 10(6) for Salmonella choleraesuis; glass retained 10(6)-10(7) organisms of all 3 test strains; porcelain retained about that amount of S. aureus but 10(5)-10(6) P. aeruginosa and 10(3)-10(4) S. choleraesuis. These data suggest that disinfectants are not similarly challenged with the AOAC-recommended test bacteria and that an alternative method should be considered to ensure comparable numbers of bacteria on penicylinders.  相似文献   

3.
Stainless steel penicylinders inoculated separately with test bacteria (Salmonella choleraesuis, Pseudomonas aeruginosa, or Staphylococcus aureus) are used in the AOAC use-dilution method (UDM) for disinfectant efficacy testing. Numbers of bacteria remaining on penicylinders were quantitatively assessed to determine if cells are washed from the penicylinders after a 10 min exposure to phosphate buffer dilution water (PBDW). Inoculated penicylinders were also examined by scanning electron microscopy (SEM) to determine the presence of cells remaining attached to the penicylinders after a 10 min exposure to a quaternary ammonium disinfectant and separately to PBDW. The percentage of cells washed from inoculated penicylinders exposed to PBDW was 89.9 for Salmonella choleraesuis, 48.8 for Pseudomonas aeruginosa, and 38.8 for Staphylococcus aureus. Qualitative examination of penicylinders by scanning electron microscopy confirmed the attachment of S. aureus and P. aeruginosa cells to penicylinders exposed separately to PBDW and a quaternary ammonium disinfectant. Few S. choleraesuis cells were observed on penicylinders exposed to PBDW and no cells were observed after disinfectant exposure. The variability of the numbers of viable cells entering the recovery media among the 3 UDM test bacteria due to cell detachment could be a significant factor in the recognized variability of the use-dilution method.  相似文献   

4.
An initial collaborative study of the AOAC use-dilution method (UDM), used for bactericidal disinfectant efficacy testing, demonstrated extreme variability of test results among the 18 laboratories testing identical hospital disinfectants. In an effort to improve the method, 32 changes were made by the UDM Task Force. These changes represented improvements in quality assurance practices and elimination of method variability; however, the basic framework of the method was retained. A second collaborative trial was conducted to determine if the interlaboratory variability of test results could be reduced to an acceptable level using the modified UDM. Twelve of the original 18 laboratories participated in the second study. Each laboratory processed 60 penicylinders (P) for each of the 6 randomly selected, federally registered disinfectants and 3 test organisms (Staphylococcus aureus, Salmonella choleraesuis, Pseudomonas aeruginosa). The number of positive penicylinders (greater than 1 positive P/60 replicates = failure) for the 6 products when P. aeruginosa was used as the challenge organism ranged 1-30, 0-36, 0-15, 0-5, 0-3, and 0-60 for the 3 quaternaries and 3 phenolics, respectively. The results of the variance components analysis for P. aeruginosa and the other 2 organisms showed that the variance components for laboratories were not significantly reduced for any organism in this study. Such interlaboratory variability of results questions the use of the original or the modified UDM for registration purposes.  相似文献   

5.
Sulfur (S) deficiencies in grain and forage crops have been detected in many agricultural regions of the world, but soil tests are not commonly used as the basis for S fertilizer recommendation programs. Errors of measurements of soil sulfate were determined to assess whether the variation among and within soil-testing laboratories could be a factor that prevent the adoption of soil testing to assess soil sulfate availability. Subsamples of 10 selected soils (Mollisols) from the Pampas (Argentina) were sent in two batches to five soil-testing laboratories. Laboratories were unaware of the existence of subsamples and performed routine sulfate analysis as if these soils came from 60 different fields. Soil sulfate ranged from 3.3 to 20.6 mg kg?1. One laboratory reported sulfate values greater than the other ones, having a mean bias of 4.1 mg kg?1 S sulfate (SO4). The other four laboratories reported similar sulfate values when soils had low sulfate availability (less than 10 mg S kg?1), even when they used different extractants. Considering only these four laboratories, average interlaboratory coefficients of variations ranged from 6 to 24% for the 10 soils. Within-laboratory mean coefficients of variation (CVs) ranged from 12 to 22%. However, mean absolute errors of all laboratories were less than 1.2 mg kg?1 S-SO4. Two laboratories reported different sulfate values for the two batches of shipment (an average difference of 4.7 and 3.8 mg kg?1 of S-SO4). Laboratories using different extractants obtained similar results, suggesting that using the same extractant is not a prerequisite to standardize laboratory results in these soils. Differences between laboratories in our study were smaller than in other interlaboratory comparisons for soil sulfate. These differences could be easily detected and corrected if laboratories participate in an interlaboratory control system. The observed low mean absolute errors suggested that, in general, all laboratories achieve acceptable precision when evaluating within the same batch of determinations. Differences between batches of shipment (within laboratory error) stressed the importance of using reference material for internal quality control.  相似文献   

6.
Net protein ratio data: AACC-ASTM collaborative study   总被引:1,自引:0,他引:1  
Seven- and 14-day net protein ratio (NPR) data were obtained from 7 laboratories for 6 protein sources: ANRC casein, lean beef, lactalbumin, textured vegetable protein, and peanut flour were fed as 10% protein (N X 6.25) in the test diet. Wheat flour, casein, and textured vegetable protein were fed as 6% protein (N X 6.25) in the test diet. Weighed dry ingredients for each diet were sent to each collaborator , who mixed the dry ingredients, then added specified amounts of corn oil and water and mixed each complete diet thoroughly. Rats were adapted for 0, 2, or 4 days, and then were fed the test diets for 28 days for protein efficiency ratio (PER) diets. The animal weight gain and feed consumption data obtained after 7 or 14 days of feeding were used to calculate NPR values. Analyses of data were done before [net protein ratio (NPR)] and after (R-NPR [relative-NPR]) adjustment of the data from each laboratory by its results for the reference protein casein. From the analysis of variance for NPR, significant (P less than 0.05) interactions were observed among laboratories, protein sources, and adaptation times of the animals (0, 2, or 4 days). Inter- and intralaboratory variability were decreased by use of 14-day values compared with 7-day values. Adjustment of the NPR data to R-NPR did not lower the intralaboratory variability but did lower the interlaboratory variability of the data. Increasing adaptation time did not consistently decrease interlaboratory or intralaboratory variability or decrease coefficients of variation (CV) of R-NPR values.(ABSTRACT TRUNCATED AT 250 WORDS)  相似文献   

7.
A novel method named cell membrane affinity chromatography was used to screen antimicrobial peptides from Jatropha curcas . A cationic antimicrobial peptide (KVFLGLK, JCpep7) was successfully isolated and identified. Antimicrobial assays indicated that JCpep7 was active against the tested microorganisms ( Salmonella typhimurium ATCC 50013, Shigella dysenteriae ATCC 51302, Pseudomonas aeruginosa ATCC 27553, Staphylococcus aureus ATCC 25923, Bacillus subtilis ATCC 23631, and Streptococcus pneumoniae ATCC 49619) with minimal inhibitory concentration (MIC) values ranging from 24 to 64 μg/mL. The antimicrobial mechanisms based on Fourier transform infrared (FTIR) spectroscopy and transmission electron microscopy (TEM) techniques showed that JCpep7 killed microbes principally via breaking of their cell walls and membranes, followed by cell lysis. The results indicated that cell membrane affinity chromatography could be a promising approach for high-throughput screening of antimicrobial peptides from J. curcas .  相似文献   

8.
The aim of the study presented here was to gain knowledge about the vapor-phase antimicrobial activity of selected essential oils and their major putatively active constituents against a range of foodborne bacterial and fungal strains. In a first step, the vapor-phase antimicrobial activities of three commercially available essential oils (EOs)-cinnamon (Cinnamomum zeylanicum), thyme (Thymus vulgaris), and oregano (Origanum vulgare)-were evaluated against a wide range of microorganisms, including Gram-negative bacteria (Escherichia coli, Yersinia enterocolitica, Pseudomonas aeruginosa, and Salmonella choleraesuis), Gram-positive bacteria (Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, and Enterococcus faecalis), molds (Penicillium islandicum and Aspergillus flavus), and a yeast (Candida albicans). The minimum inhibitory concentrations (MICs) were generally lower for oregano EO than for the thyme and cinnamon EOs, especially against the relatively resistant Gram-negative. The persistence of the EOs' antimicrobial activities over time was assessed, and changes in the composition of the atmosphere they generated over time were determined using single-drop microextraction (SDME) in combination with gas chromatography-mass spectrometry (GC-MS) and subsequent analysis of the data by principal component analysis (PCA). More relevant chemicals were selected. In addition, the vapor-phase activities of putatively key constituents of the oils were screened against representative Gram-positive (L. monocytogenes) and Gram-negative (S. choleraesuis) bacteria, a mold (A. flavus), and a yeast (C. albicans). Of the tested compounds, cinnamaldehyde, thymol, and carvacrol showed the strongest antimicrobial effectiveness, so their MICs, defined as the minimum vapor concentrations that completely inhibited detectable growth of the microorganisms, were calculated. To check for possible interactions between components present in the EOs, cinnamon EO was fortified with cinnamaldehyde and thyme EO with thymol, and then the antimicrobial activities of the fortified oils were compared to those of the respective unfortified EOs using fractional inhibitory concentration (FIC) indices and by plotting inhibition curves as functions of the vapor-phase concentrations. Synergistic effects were detected for cinnamaldehyde on A. flavus and for thymol on L. monocytogenes, S. choleraesuis, and A. flavus. In all other cases the fortification had additive effects, except for cinnamaldehyde's activity against S. choleraesuis, for which the effect was antagonistic. Finally, various microorganisms were found to cause slight changes over time to the atmospheres generated by all of the EOs (fortified and unfortified) except the fortified cinnamon EO.  相似文献   

9.
The antimicrobial activity of essential oils (EOs) of cinnamon (Cinnamon zeylanicum), clove (Syzygium aromaticum), basil (Ocimum basillicum), rosemary (Rosmarinus officinalis), dill (Anethum graveolens), and ginger (Zingiber officinalis) was evaluated over a range of concentrations in two types of contact tests (solid and vapor diffusion). The EOs were tested against an array of four Gram-positive bacteria (Staphylococcus aureus, Bacillus cereus, Enterococcus faecalis, and Listeria monocytogenes), four Gram-negative bacteria (Escherichia coli, Yersinia enterocolitica, Salmonella choleraesuis, and Pseudomonas aeruginosa), and three fungi (a yeast, Candida albicans, and two molds, Penicillium islandicum and Aspergillus flavus). The rationale for this work was to test the possibility of creating a protective atmosphere by using natural compounds that could extend the shelf life of packaged foodstuffs while minimizing organoleptic alterations. In the solid diffusion tests, cinnamon and clove gave the strongest (and very similar) inhibition, followed by basil and rosemary, with dill and ginger giving the weakest inhibition. The fungi were the most sensitive microorganisms, followed by the Gram-positive bacterial strains. The Gram-negative strain P. aeruginosa was the least inhibited. The composition of the atmosphere generated by the EOs, and their minimum inhibitory concentrations (MICs), were determined using a disk volatilization method, in which no inhibition from rosemary or basil was observed. Cinnamon and clove, once again, gave similar results for every microorganism. As a general rule, MIC (fungi) < MIC (bacteria) with no clear differences between Gram-positive or -negative strains except for P. aeruginosa, which was not inhibited by any of the EOs in the vapor phase. The atmosphere generated from the EOs was analyzed by means of solid-phase microextraction combined with gas chromatography-ion trap mass spectrometry. Differences among the volatiles in the EOs, which may be responsible for the differences in their antimicrobial performances, were found.  相似文献   

10.
Fourier transform infrared (FT-IR) spectroscopy and multivariate analysis were used to identify Pseudomonas aeruginosa and Escherichia coli ATCC 25922 inoculated into bottled drinking water. Three inoculation treatments were examined: (i) E. coli ATCC 25922 (N = 3), (ii) P. aeruginosa (N = 3), and (iii) a 1:1 (v:v) mixed culture of both P. aeruginosa and E. coli ATCC 25922 (N = 3). The control treatment was noninoculated drinking water (N = 3). Second derivative transformation and loadings plots over the range of 1800-900 cm(-1) indicate variations in the following bacterial constituents: amide I band ca. 1650 cm(-1), amide II band ca. 1540 cm(-1), phosphodiester backbone of nucleic acids ca. 1242 and 1080 cm(-1), and polysaccharide compounds ca. 1050-950 cm(-1). Cells with the different treatments were clearly segregated from a mean centered principal component analysis. By using soft independent modeling of class analogy analysis, spectra from a given treatment could be correctly classified 83-88% of the time. These results suggest that FT-IR spectroscopy can determine whether a pure culture is present, in addition to confirming that this method can discriminate between closely related bacteria based on differences in biochemical and phenotypic characteristics that can be detected in this spectral region.  相似文献   

11.
Aliphatic (2E)-alkenals and alkanals characterized from the fresh leaves of the coriander Coriandrum sativum L. (Umbelliferae) were found to possess bactericidal activity against Salmonella choleraesuis ssp. choleraesuis ATCC 35640. (2E)-Dodecenal (C(12)) was the most effective against this food-borne bacterium with the minimum bactericidal concentration (MBC) of 6.25 microg/mL (34 microM), followed by (2E)-undecenal (C(11)) with an MBC of 12.5 microg/mL (74 microM). The time-kill curve study showed that these alpha,beta-unsaturated aldehydes are bactericidal against S. choleraesuis at any growth stage and that their bactericidal action comes in part from the ability to act as nonionic surfactants.  相似文献   

12.
A liquid chromatographic (LC) method for the determination of chloramphenicol (CAP) residues in meat at the 10 microgram/kg level was tested in an interlaboratory study. The method used, based on aqueous extraction and sample cleanup with a cartridge containing Extrelut, was published earlier. A prestudy to familiarize collaborators with the method was performed before the actual interlaboratory precision study. The meat samples used in the precision study were prepared by diluting dosed chicken and pig muscle tissues with blank tissues from other species. Fourteen laboratories received 20 meat samples; 13 laboratories actually participated in the study. Two blank samples and 2 positive samples each of pig, calf, chicken, lamb, and cow meat were tested. The chloramphenicol concentrations in the positive samples ranged from 6.5 to 21 micrograms/kg. The overall mean reproducibility coefficient of variation was 17.9% after the results per laboratory were corrected for the mean recovery obtained within each sample series. The overall mean recovery was 55.1% with a coefficient of variation of 18.0% at the 10 micrograms/kg level. The limit of detection, based on chromatograms of blank samples, was estimated to be 1.5 micrograms/kg of chloramphenicol. No false positives or false negatives were observed in the concentration range tested; only 2 false positive results above the detection limit (1.7 and 6 micrograms/kg) on a total number of 60 blank analyses (3.3%) were observed.  相似文献   

13.
Enumeration of Staphylococcus aureus in foods was collaboratively studied by comparing the present AOAC final action method, 46.062, which uses trypticase soy broth with 10% NaCl to a proposed replacement method which uses the same broth with 1% sodium pyruvate added. Fifteen collaborators analyzed uninoculated samples of milk, tuna salad, and ground turkey, as well as samples inoculated with low (10(2) cells/g), middle (10(4) cells/g), and high (10(6) cells/g) levels of S. aureus. The samples were frozen immediately to maintain the inoculated level of S. aureus in the food. A different strain of S. aureus was used for each food; heat-stressed S. aureus cells were used to inoculate the milk samples. The pyruvate-amended broth significantly (alpha = 0.05) increased enumeration of low, middle, and high levels of S. aureus from milk and ground turkey, and from tuna salad at middle and high levels. The pyruvate-amended media method has been adopted official first action to replace method 46.062.  相似文献   

14.
The International Dairy Federation develops and studies methods for the analysis of milk and milk products. A first draft of an IDF protocol for conducting interlaboratory studies is presented. Major features of the protocol are type and number of participating laboratories; nature, duplication, and number of sample materials; final design (numbers of samples and laboratories); statistical analysis of the data; report of the final results.  相似文献   

15.
The U.S. Food and Drug Administration (FDA) sponsored an interlaboratory study of a liquid chromatographic determinative procedure for lasalocid sodium in chicken skin with adhering fat. Four laboratories analyzed 35 dosed tissue samples and 82 fortified tissue samples containing lasalocid at levels ranging from 0.1 to 0.6 ppm. Samples were homogenized with acetonitrile, washed with hexane, and partitioned into the mobile phase prior to analysis liquid chromatography with fluorescence detection. The results of the interlaboratory study showed good reproducibility for fortified samples. Fortification levels, average recoveries, and interlaboratory percent coefficients of variation were as follows: 0.6 ppm, 0.57 ppm, and 9.7; 0.3 ppm, 0.25 ppm, and 9.1; and 0.15 ppm, 0.14 ppm, and 7.0, respectively. Data for analysis of the dosed tissue also showed good agreement among the laboratories.  相似文献   

16.
Chitin neoglycoconjugates (BSA-CO) were obtained by the conjugation of bovine serum albumin (BSA) with chitin oligosaccharides (CO) through the Maillard reaction (nonenzymatic glycation). CO produced by acid hydrolysis of chitin were fractionated using an ultrafiltration membrane system (1-3 kDa cutoff). The Maillard reaction was carried out by heating a freeze-dried mixture containing BSA and CO at 60 °C (under 43% relative humidity for 6 and 12 h). BSA-CO were characterized by available amino groups content, intrinsic tryptophan emission spectra, gel electrophoresis, and mass spectrometry. Biological assays included interaction with wheat germ agglutinin (WGA) and with bacterial adhesins of Escherichia coli K88+ and Salmonella choleraesuis. Glycation of BSA was revealed by reduction of available amino groups and fluorescence intensity and also retarded migration through SDS-PAGE. Conjugation of BSA with chitin oligomers appeared to be time dependent and was confirmed by mass spectrometry, by which molecular mass increase for monomers and dimers was observed. Monomers were estimated to contain either one or two glycation sites (at 6 and 12 h of treatment, respectively), with one or two tetrasaccharide units attached. Consequently, dimers showed two or four glycation sites. BSA-CO presented biological recognition by WGA and E. coli K88+ and S. cholerasuis adhesins. The strategy used in this work represents a simple method to obtain glycoconjugates to study applications involving protein-carbohydrate recognition.  相似文献   

17.
A liquid chromatographic (LC) method previously published for the determination of carbadox in finished feeds and premixes was slightly modified and tested in an interlaboratory study. The feed samples are extracted with methanol-acetonitrile (50 + 50) after wetting with water. The extracts are purified over a short alumina column. An aliquot of the eluate is analyzed with reverse phase liquid chromatography with ultraviolet detection. Before the actual interlaboratory study, a prestudy with 2 familiarization feed samples was performed. For the interlaboratory study, 2 series of meal and pelleted samples were prepared with carbadox from different suppliers. Eight collaborating laboratories received 6 feed samples previously milled and ground and 4 pelleted samples which had to be ground by the collaborator's in-house method. Collaborators also received 3 carbadox concentrates (about 10% w/w) and 4 premix samples derived from the concentrates (about 1% w/w). Coefficients of variation under reproducibility conditions were 8.3% for meal samples and 4.9% for pellets. A minor but significant effect was noted for the influence of pelleting temperature on the carbadox content. A minor and insignificant effect was observed for the influence of the milling and grinding procedure on the carbadox content. Alumina cleanup of 1% premixes was not essential, although the resulting chromatograms were cleaner. A slight difference in reproducibility was observed with concentrates (10%) when 0.2 or 0.5 g sample size was used, although the average carbadox concentration found was the same. For premixes and concentrates, coefficients of variation under reproducibility conditions were low, ranging from 2.9 to 7.5%.  相似文献   

18.
The inhibitory activity of Curcuma longa L. (turmeric) rhizome constituents against sortase A, a bacterial surface protein anchoring transpeptidase, from Staphylococcus aureus ATCC 6538p was evaluated. The activity of the isolated compounds (1-4) was compared to that of the positive control,p-hydroxymecuribenzoic acid (pHMB). The biologically active components of C. longa rhizome were characterized by spectroscopic analysis as the curcuminoids curcumin (1), demethoxycurcumin (2), and bisdemethoxycurcumin (3). Curcumin was a potent inhibitor of sortase A, with an IC50 value of 13.8 +/- 0.7 microg/mL. Bisdemethoxycurcumin (IC50 = 31.9 +/- 1.2 microg/mL) and demethoxycurcumin (IC50 = 23.8 +/- 0.6 microg/mL) were more effective than pHMB (IC50 = 40.6 +/- 1.2 microg/mL). The three isolated compounds (1-3) showed no growth inhibitory activity against S. aureus strain Newman, with minimum inhibitory concentrations (MICs) greater than 200 microg/mL. Curcumin also exhibited potent inhibitory activity against S. aureus cell adhesion to fibronectin. The suppression of fibronectin-binding activity by curcumin highlights its potential for the treatment of S. aureus infections via inhibition of sortase activity. These results indicate that curcumin is a possible candidate in the development of a bacterial sortase A inhibitor.  相似文献   

19.
A joint U.S. Environmental Protection Agency/AOAC interlaboratory method validation study was conducted on EPA Method 508, Determination of Chlorinated Pesticides in Water by Gas Chromatography with an Electron Capture Detector, to determine the mean recovery and precision for analyses of 29 pesticides in reagent water and finished drinking water. The study design was based on Youden's nonreplicate plan for collaborative tests of analytical methods. The waters were spiked with 29 pesticides at 6 concentration levels, as 3 Youden pairs. Eleven volunteer laboratories extracted the spiked test waters with methylene chloride, performed a solvent exchange with methyl tert-butyl ether, and analyzed an aliquot of each extract by gas chromatography with electron capture detection. Results were analyzed using an EPA computer program, interlaboratory Method Validation Study (IMVS), which measured recovery and precision for each of the 29 pesticides and compared the performance of the method between water types. Method 508 was judged acceptable for all analytes tested. Only 3 analytes (alpha-chlordane, 4,4'-DDE, and methoxychlor) exhibited practical significant matrix effects. The method has been adopted official first action.  相似文献   

20.
Comparison of stainless steel penicylinders used in disinfectant testing   总被引:2,自引:0,他引:2  
Two brands of stainless steel penicylinders, S&L Metal Products and Fisher Scientific, were simultaneously tested to determine if they provide comparable results when used in the AOAC use-dilution method of disinfectant testing. Results showed consistently more positive tubes for the Fisher brand of penicylinders than for the S&L, regardless of the surface finish of the test cylinders.  相似文献   

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