3M Petrifilm Staph Express Count plate method for the enumeration of Staphylococcus aureus in selected dairy foods: collaborative study

The 3M Petrifilm Staph Express Count plate method was compared with AOAC Official Method 975.55 for the enumeration of Staphylococcus aureus in selected foods. Five foods--ice cream, raw milk, yogurt, whey powder, and cheese--were analyzed for S. aureus by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample, a low inoculation level, a medium inoculation level, and a medium inoculation level with background flora, each in duplicate. The mean log10 counts for the methods were comparable for all 5 foods. The repeatability and reproducibility variances of the 24 h Petrifilm Staph Express Count plate method were similar to those of the 72 h standard method.     

3M Petrifilm Staph Express Count plate method for the enumeration of Staphylococcus aureus in selected types of processed and prepared foods: collaborative study

The 3M Petrifilm Staph Express Count plate method was compared with AOAC Official Method 975.55 for the enumeration of Staphylococcus aureus in selected foods. Five foods--frozen lasagna, custard, frozen mixed vegetables, frozen hashbrowns, and frozen batter-coated mushrooms--were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample, a low inoculation level, a medium inoculation level, and a medium inoculation level with background flora, each in duplicate. The mean log10 counts for the methods were comparable for all 5 foods. The repeatability and reproducibility variances of the 24 h Petrifilm Staph Express Count plate method were similar to those of the 72 h standard method.     

Comparison of Petrifilm Staph Express Count System with the bacteriological analytical manual direct-plating method for enumeration of Staphylococcus aureus in artificially contaminated hard cheese

The 3M Petrifilm Staph Express Count System was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual (BAM) direct-plate count method for the enumeration of Staphylococcus aureus in 6 types of artificially contaminated hard cheese (Asiago, Cheddar, Gruyère, Parmesan, Romano, and Swiss). Five different samples of each cheese type were inoculated with S. aureus (ATCC 25923) to achieve low, medium, and high inoculum levels. S. aureus was enumerated by the Petrifilm and BAM methods, and the results were compared. Multivariate analysis of variance revealed no significant differences (P<0.05) between the 2 methods. The Petrifilm method compared favorably with the BAM procedure. The rapid method was more convenient to use, considerably faster, and less expensive to perform than the BAM method.     

Dry rehydratable film method for enumerating confirmed Escherichia coli in poultry, meats, and seafood: collaborative study

A rehydratable dry-film plating method for Escherichia coli, the Petrifilm E. coli/Coliform (EC) Count Plate in foods, has been compared with the AOAC INTERNATIONAL most probable number (MPN) method. Eleven laboratories participated in the collaborative study. Three E. coli levels in 8 samples each of frozen raw ground turkey, frozen raw ground beef, and frozen cooked fish were tested in duplicate. Mean log counts for the Petrifilm plate procedure were not significantly different from those for the MPN procedure for cooked fish samples inoculated with low or high inocula levels, for samples of raw turkey inoculated at medium level, and for beef inoculated at low, medium, and high levels. Repeatability and reproducibility variances of the Petrifilm EC Plate method recorded at 24 h were as good as or better than those of the MPN method. The dry rehydratable film method for enumerating confirmed E. coli in poultry, meats, and seafood has been adopted first action by AOAC INTERNATIONAL.     

Dry rehydratable film method for rapid enumeration of coliforms in foods (3M Petrifilm Rapid Coliform Count plate): collaborative study

A rehydratable dry-film plating method for coliforms in foods, the 3M Petrifilm Rapid Coliform Count plate method, was compared with the U.S. Food and Drug Administration's Bacteriological Analytical Manual method for nondairy foods and the American Public Health Association's Standard Methods for the Examination of Dairy Products (SMEDP) method for dairy foods. Six food types, vanilla ice cream, cheddar cheese, fresh refrigerated uncooked pasta, wheat flour, prepared frozen macaroni and cheese, and frozen hash browns, were analyzed for coliforms by 11 collaborating laboratories. For each food product tested, the collaborators received 8 blind samples consisting of a control sample and 3 levels of inoculated sample, each in duplicate. The mean log counts for the methods were comparable. The repeatability and reproducibility variances of the Petrifilm Rapid Coliform Count method at 14 and 24 h were not significantly different from those of the standard methods.     

Petrifilm rapid S. aureus Count Plate method for rapid enumeration of Staphylococcus aureus in selected foods: collaborative study

A rehydratable dry-film plating method for Staphylococcus aureus in foods, the 3M Petrifilm Rapid S. aureus Count Plate method, was compared with AOAC Official Method 975.55 (Staphylococcus aureus in Foods). Nine foods-instant nonfat dried milk, dry seasoned vegetable coating, frozen hash browns, frozen cooked chicken patty, frozen ground raw pork, shredded cheddar cheese, fresh green beans, pasta filled with beef and cheese, and egg custard-were analyzed for S. aureus by 13 collaborating laboratories. For each food tested, the collaborators received 8 blind test samples consisting of a control sample and 3 levels of inoculated test sample, each in duplicate. The mean log counts for the methods were comparable for pasta filled with beef and cheese; frozen hash browns; cooked chicken patty; egg custard; frozen ground raw pork; and instant nonfat dried milk. The repeatability and reproducibility variances of the Petrifilm Rapid S. aureus Count Plate method were similar to those of the standard method.     

Evaluation of a dry, rehydratable film method for rapid enumeration of Staphylococcus aureus

Results with the new 3M Petrifilm Rapid S. aureus Count (RSA) Plate method were compared with those of the classical Baird-Parker agar (BPA) method for detection and enumeration of Staphylococcus aureus. Studies on 219 bacterial strains demonstrated that the Petrifilm RSA plate is more sensitive than and as specific as the classical BPA method for confirmed identification of S. aureus. Counts of colonies from 71 pure cultures, 61 naturally contaminated food samples, and more than 750 artificially inoculated food samples showed that the Petrifilm RSA method was as effective as the classical BPA method for identification and enumeration of S. aureus. The Petrifilm RSA method gave results in one-third the time required for the classical method.     

Comparative evaluation of the association among enumeration methods and production of enterotoxins in food-derived Staphylococcus aureus

Staphylococcal food poisoning is one of the most common foodborne diseases worldwide; it results from the ingestion of staphylococcal enterotoxins (SEs) in food, mainly Staphylococcus aureus. This study investigated the statistical relationships among morphological enumerations of food-derived S. aureus and production of SEs using different methodologies. Food samples naturally contaminated with coagulase-positive S. aureus were submitted for enumeration on Baird-Parker (BP) agar, Rabbit Plasma Fibrinogen agar (RPFA), and Petrifilm Staph Express count system (STX), and the morphologically typical colonies were isolated for VIDAS and real-time (RT) PCR tests. RPFA and STX displayed better performance for the enumeration of SE-positive S. aureus when compared with BP, including higher frequencies of SE-positive isolates and better correlation indices between typical and SE-positive counts. Among all the evaluated culture media, no significant difference (P > 0.05) was shown on the frequencies of typical colonies that carried 11 individual se genes. In addition, results for SE identification between VIDAS and RT-PCR assay were unparalleled. These data will be valuable for the selection of methods for inspection of food-derived S. aureus.     

3M Petrifilm enterobacteriaceae count plate method for enumeration of enterobacteriaceae in selected foods: collaborative study

The practice of detecting and enumerating all oxidase-negative, glucose-fermenting-Gram-negative rods (i.e., the family Enterobacteriaceae) is used to indicate unsanitary or inadequate food processing conditions. The objective of this interlaboratory collaborative study was to evaluate and compare the methods described in Standard Methods for the Examination of Dairy Products (SMEDP) and the Compendium of Methods for the Microbiological Examination of Foods (Compendium) with a commercial product, the 3M Petrifilm Enterobacteriaceae Count Plate, for the recovery of Enterobacteriaceae in foods. Six foods--cheddar cheese, milk, flour, frozen prepared meals, frozen broccoli, and nut pieces--were analyzed for Enterobacteriaceae by 12 collaborating laboratories. For each food tested, the collaborators received 8 blind test portions consisting of a control test portion and 3 levels of inoculated test portion, each in duplicate. Each test portion was tested by the Petrifilm Enterobacteriaceae Count Plate method as well as the SMEDP or Compendium methods. The precision estimates (repeatability or within-laboratory variation, and reproducibility or between-laboratory variation) were calculated with standard statistical techniques.     

Sensitivity and specificity of the Sanita-kun Aerobic Count: internal validation and independent laboratory study

The Sanita-kun Aerobic Count consists of a transparent cover film, an adhesive sheet, a layer of nonwoven fabric, and a water-soluble compound film, including a culture medium formula for detection of aerobic microorganisms. The Sanita-kun sheet was validated for 14 food categories in an internal study and an independent study was conducted on ground beef and hot dogs. Both studies showed no significant difference in performance between 5 or 8 replicates of the Sanita-kun sheets and AOAC Method 966.23, excluding some lots of foods. The correlation coefficient to plate count agar in the internal accuracy study was 0.99. The average relative standard deviation for repeatability of total foods was 0.26 and 0.19, respectively, excluding < 10 average counts. The ruggedness study, which examined the influence of incubation temperature and period, recommended incubation of the Sanita-kun sheet at 32.5 +/- 2.5 degrees C for 46 +/- 2 h. Comparison of 3 lots of Sanita-kun sheets showed no decrease of performance in the older lot. The shelf-life of the sheet is at least 14 months. The Sanita-kun Aerobic Counts has been granted AOAC Performance Tested Method status.     

Validation of RAPID'Staph for enumeration of Staphylococcus aureus in foods. Performance-Tested Method

RAPID'Staph (Bio-Rad Laboratories, Hercules, CA) is a medium for differentiation and enumeration of coagulase-positive Staphylococcus aureus in food. RAPID'Staph medium is based on a Baird Parker formula optimized for the detection and enumeration of S. aureus in 24 h. The principle of the medium relies on the capacity of S. aureus to reduce tellurite (production of black colonies) and to provoke proteolysis of egg yolk (production of clear halo around the colony). Four foods (pasteurized whole milk, custard pie, processed ham, and smoked salmon) were selected to compare the performance of RAPID'Staph agar to AOAC Official Method 975.55. Method comparison studies demonstrated excellent agreement between the methods. Inclusivity and exclusivity rates of the medium were 100%. RAPID'Staph agar performed as expected when minor procedural variations were introduced, validating the ruggedness of the method. There was no difference in performance between the dehydrated and ready-to-use formulations of the media.     

Comparison of the compact dry EC with the most probable number method (AOAC official method 966.24) for enumeration of Escherichia coli and coliform bacteria in raw meats. Performance-Tested Method 110402

Compact Dry E. coli/Coliform Count (EC) is a ready-to-use test method for the enumeration of Escherichia coli and coliform bacteria in food. The plates are presterilized and contain culture medium and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL diluted sample inoculated onto the center of the self-diffusible medium, allowing the solution to diffuse by capillary action. The plate can be incubated at 35 degrees C for 20-24 h and the colonies counted without any further working steps. The Compact Dry EC medium plates were validated as an analysis tool for determining colony-forming units (CFU) of E. coli and coliform bacteria from a variety of raw meats using 5 different types of raw meats. The performance tests were conducted at 35 degrees C. In all studies performed, no apparent differences were observed between the Compact Dry EC method and the AOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.93 (E. coli) and r2 = 0.93 (coliform bacteria) could be assigned, as stated in the application for "Performance-Tested Method."     

Evaluation of the VIDAS staph enterotoxin II (SET 2) immunoassay method for the detection of staphylococcal enterotoxins in selected foods: collaborative study

A multilaboratory study was conducted to determine the limit of detection (LOD) of Staphylococcal enterotoxins (SET) in 5 foods. Cooked chicken, ham, potato salad, pasteurized liquid whole milk, and canned mushrooms were each spiked with a different enterotoxin (A, B, C1, D, or E), and tested at 0.25 and 0.5 ng/g SET levels to determine the LOD of the assay for those foods in a collaborative study. Unspiked controls were also included. A total of 19 laboratories representing government and industry participated. In this study, 1674 test portions were analyzed, of which 1638 were used in the statistical analysis. Of the 1638 test portions used in the statistical analysis, 1104 were spiked test portions, of which 1073 were positive by the VIDAS Staph enterotoxin II (SET 2) method. The detection rates at the 0.25 ng/mL level were cooked chicken, 98.2%; ham, 99.0%; potato salad, 99.1%; liquid whole milk, 85.2%; and canned mushrooms, 100%. The detection rates at the 0.5 ng/mL level were cooked chicken, 97.4%; ham, 98.1%; potato salad, 100%; liquid whole milk, 99.0%; and canned mushrooms, 100%. The data indicate that the SET 2 method is capable of detecting SET at 0.25 ng/g in cooked chicken, ham, potato salad, and canned mushrooms and at 0.5 ng/g in pasteurized liquid whole milk.     

Equivalence of Visual Immunoprecipitate Assay (VIP) for Salmonella for the detection of motile and nonmotile Salmonella in all foods to AOAC culture method: collaborative study

Six foods representative of a wide variety of processed, dried powder processed, and raw food types were analyzed by the Visual Immunoprecipitate Assay (VIP) for Salmonella and AOAC INTERNATIONAL culture method. Paired samples of each food type were simultaneously analyzed; one sample by the VIP method and one by the AOAC culture method. A total of 24 laboratories representing federal government agencies and private industry, in the United States and Canada, participated in this collaborative study. Food types were inoculated with species of Salmonella with the exception of raw ground chicken, which was naturally contaminated. No statistical differences (p < 0.05) were observed between VIP for Salmonella interpretation and the AOAC culture method for any inoculation level of any food type or naturally contaminated food. The method was adopted Official First Action status by AOAC INTERNATIONAL.     

Validation of the Hygicult E dipslides method in surface hygiene control: a Nordic collaborative study

A collaborative study with Enterobacteriaceae was conducted to validate Hygicult E dipslides by comparison with violet red bile glucose agar (VRBGA) contact plates and swabbing, using stainless steel surfaces artificially contaminated with microbes at various levels. Twelve laboratories participated in the validation procedure. The total number of collaborative samples was 108. The microbial level in each sample was assessed in triplicate by using the 3 above-mentioned methods. No Enterobacteriaceae were used at the low inoculation level. At the middle inoculation level, the percentages detached from the test surfaces were 16.6 with the Hygicult E method, 15.3 with the contact plate method, and 14.6 with swabbing; at the high innoculation level, the percentages were 14.5, 15.8, and 9.8, respectively. The percentage of acceptable results after the removal of outliers was 97.2. Repeatability relative standard deviations ranged from 33.4 to 44.9%; reproducibility relative standard deviations ranged from 45.2 to 77.1%. The Hygicult E dipslide, VRBGA contact plate, and swabbing methods gave similar results at all 3 microbial levels tested: <1.0 colony-forming units (CFU)/cm2 at the low level, 1.2-1.3 CFU/cm2 at the middle level (theoretical yield 8.0 CFU/cm2), and 1.2-2.0 CFU/cm2 at the high level (theoretical yield 12.5 CFU/cm2).     

Evaluation of the VIDAS Listeria (LIS) immunoassay for the detection of Listeria in foods using demi-Fraser and Fraser enrichment broths, as modification of AOAC Official Method 999.06 (AOAC Official Method 2004.06)

A multilaboratory study was conducted to compare the VIDAS LIS immunoassay with the standard cultural methods for the detection of Listeria in foods using an enrichment modification of AOAC Official Method 999.06. The modified enrichment protocol was implemented to harmonize the VIDAS LIS assay with the VIDAS LMO2 assay. Five food types--brie cheese, vanilla ice cream, frozen green beans, frozen raw tilapia fish, and cooked roast beef--at 3 inoculation levels, were analyzed by each method. A total of 15 laboratories representing government and industry participated. In this study, 1206 test portions were tested, of which 1170 were used in the statistical analysis. There were 433 positive by the VIDAS LIS assay and 396 positive by the standard culture methods. A Chi-square analysis of each of the 5 food types, at the 3 inoculation levels tested, was performed. The resulting average Chi square analysis, 0.42, indicated that, overall, there are no statistical differences between the VIDAS LIS assay and the standard methods at the 5% level of significance.     

依诺沙星与铁配合物的合成、表征及其生物活性

由水热法合成了依诺沙星铁配合物,用固体培养基平板药敏试验方法测定了配合物的体外抗菌活性。结果表明,合成的依诺沙星铁配合物对金黄色葡萄球菌、肺炎链球菌、绿脓杆菌、白色念珠菌都具有抗菌活性,但只有对金黄色葡萄球菌和绿脓杆菌的抗菌活性强于配体本身。     

CuCl2 heterogenized on metformine‐modified polystyrene resin as an antibacterial agent and recyclable nanocatalyst for Ullmann‐type C‐N coupling reactions

Merrifield Resin was functionalized with metformine and applied as a solid support to immobilize the CuCl₂. The Ps‐Met/CuCl₂ was characterized by several techniques including Fourier transform infrared (FTIR), field emission scanning electron microscopy (FESEM), energy‐dispersive X‐ray spectroscopy (EDX), wavelength‐dispersive X‐ray spectroscopy (WDX) and inductively coupled plasma (ICP). The Ps‐Met/CuCl₂ was used as an efficient recyclable solid nanocatalyst for N‐arylation of indole and aniline through coupling reactions of Ullmann‐type C‐N. The advantages of this method are easy workup, improved yields, and simple recovery via filtration. Ultimately, the Ps‐Met/CuCl₂ antibacterial property was examined against two bacteria (Staphylococcus aureus (Staph. aureus) and Escherichia coli (E. coli)) and indicated its antibacterial performance against gram negative (E. coli) bacteria and gram positive (Staph. aureus).     

Equivalence of assurance Gold Enzyme Immunoassay for visual or instrumental detection of motile and nonmotile Salmonella in all foods to AOAC culture method: collaborative study

Six foods representative of a wide variety of processed, dried powder processed, and raw food types were analyzed by the Assurance Gold Salmonella Enzyme Immunoassay (EIA) and AOAC INTERNATIONAL culture method. Paired samples of each food type were simultaneously analyzed; one sample by the Assurance method and one by the AOAC culture method. The results for Assurance method were read visually and instrumentally with a microplate reader. A total of 24 laboratories representing federal government agencies and private industry, in the United States and Canada, participated in this collaborative study. Food types were inoculated with species of Salmonella with the exception of raw ground chicken, which was naturally contaminated. No statistical differences (p < 0.05) were observed between Assurance Gold Salmonella EIA with either visual or instrumental interpretation and the AOAC culture method for any inoculation level of any food type or naturally contaminated food. The Assurance visual and instrumental options of reading sample reactions produced the same results for 1277 of the 1296 sample and controls analyzed.     

Antimicrobial Property of Nanosilver Colloid Prepared by Electrical Spark Discharge Method on <Emphasis Type="Italic">Aspergillus niger</Emphasis>

This study employed an electrical spark discharge method (ESDM) to prepare a nano-Ag colloid as an antifungal solution. The solution was diluted to two concentrations, and the fungal medium prepared in this study was coated with Aspergillus niger. The nano-Ag colloid solution was mixed with A. niger in various concentrations and dripped onto 3M Petrifilm plates. Inhibited growth observed after several days confirmed the antifungal effect of the nano-Ag colloid on A. niger. Because direct washing produced inaccurate quantitation and yielded A. niger in an excessively high concentration, this study employed an inoculation loop method for A. niger quantitation. The concentrations of A. niger ranged from 10^?2 to 10^?7%. The optimal colony count was observed on day 2. During an experiment regarding the antifungal effect of the ESDM-prepared nano-Ag colloid on A. niger, 3M Petrifilm plates were employed to observe the growth of A. niger. The colony count of 10^?2% A. niger without nano-Ag colloid was approximately 60. After the nano-Ag colloid was added, the colony count substantially decreased to approximately 10. The colony count of 10^?7% A. niger was reduced to 11 or lower after the nano-Ag colloid was added. The results confirmed the antifungal effect of the nano-Ag colloid on the growth of A. niger.