Evaluation of a new chromogenic agar medium for Alicyclobacillus acidoterrestris.

Spoilage of fruit juices by a thermoacidophilic spore-forming bacterium, Alicyclobacillus acidoterrestris, is a big problem for fruit juice industries worldwide. We have developed a novel chromogenic selective agar medium (EAATSM) for the isolation and enumeration of A. acidoterrestris. A. acidoterrestris strains appeared as blue colonies on the EAATSM. Other Alicyclobacillus strains appeared as white colonies or were inhibited. A study comparing EAATSM and YSG agar was carried out using artificially contaminated samples of 50 fruit juice products. The correlation coefficient between EAATSM and YSG was 0.991.

Evaluation of the dry sheet medium (Compact Dry ETB(R)) for enumerating Enterobacteriaceae in meat samples.

Compact Dry ETB(R) (CD-ETB, ETB; Enterobacteriaceae) , a ready-to-use and self-diffusible dry medium sheet culture system for the detection and enumeration of Enterobacteriaceae, was evaluated. A total of 35 Enterobacteriaceae strains, which were studied for the inclusivity study, grew and formed red-purple colored colonies on the CD-ETB(R). When 17 gram-negative bacteria other than Enterobacteriaceae were inoculated for the exclusivity study, 2 strains grew and formed red-purple colonies, 9 strains formed colorless colonies, and 6 strains failed to grow. A total of 43 gram-positive bacteria and 3 yeasts failed to grow. The CD-ETB method was compared with the Violet Red Bile Glucose (VRBG) agar method according to ISO 21528-2 and the 3M Petrifilm(TM) EB (3M-EB(R)) method in 53 meat samples. The correlation coefficients between CD-ETB(R) and VRBG agar, CD-ETB(R) and 3M-EB(R), and 3M-EB(R) and VRBG agar were 0.962, 0.992 and 0.960, respectively.

Compact Dry(R) X-BC for the enumeration of Bacillus cereus in food samples.

We evaluated the effectiveness of using Compact Dry(R) X-BC (CD-XBC), a ready-to-use and self-diffusing dry medium sheet culture system based on a novel detection principle, for the detection and enumeration of Bacillus cereus. All 13 B. cereus strains, which were studied for the inclusivity study, grew as blue/green colonies on the CD-XBC. When 3 yeast strains and 103 bacterial strains other than B. cereus were tested for the exclusivity study, 5 strains formed white colonies, and 4 strains formed blue/green colonies, while 94 other strains failed to grow. The 4 strains that formed blue/green colonies were B. thuringiensis, which is known to have the same biochemical features as B. cereus. The CD-XBC method was compared with the MYP agar method (MYP) and the NGKG agar method (NGKG) in 130 artificially contaminated food samples. The correlation coefficients between CD-XBC and MYP, and CD-XBC and NGKG were 0.972 and 0.971, respectively.

Evaluation of a novel rapid detection system for yeasts and molds.

A novel system, the NISSUI rapid detection system, has been developed to rapidly detect yeasts and molds in food. This system consists of a liquid medium containing resazurin as a redox indicator, a unique original micro-well dish containing 676 micro-wells, and a fluorescence-monitoring instrument with an incubator. To evaluate this system, orange juice, milk, and physiological saline solutions artificially inoculated with yeasts or molds were used as samples. Comparison of the new system used at 28ºC for 48 h with a spread-plate method using a potato-dextrose-agar plate at 25ºC for 120 h yielded a correlation coefficient (r) of 0.95. Our data reveal that the new method considerably shortens the time required for detection of yeasts and molds in food.

Evaluation of the compact dry X-SA method for enumerating Staphylococcus aureus in artificially contaminated food samples.

Compact Dry X-SA (CD-XSA), a ready-to-use and self-diffusible dry medium sheet culture system for the detection and enumeration of Staphylococcus aureus, was evaluated. A total of 50 S. aureus strains, which were studied for the inclusivity study, grew as blue-colored colonies on the CD-XSA. When 114 bacteria other than S. aureus and 3 yeasts were inoculated for the exclusivity study, 37 strains produced white colonies, and 4 strains produced blue colonies, and 3 strains produced magenta colonies, while 73 other strains failed to grow. The CD-XSA method was compared with the mannitol salt agar with egg yolk (MSEY) method, the Baird-Parker agar (BP) method and the 3M Petrifilm(TM) STX (3M-STX) method in 105 artificially contaminated food samples. The correlation coefficients between CD-XSA and MSEY, CD-XSA and BP, and CD-XSA and 3M-STX were 0.945, 0.960 and 0.977, respectively.

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

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

Evaluation of the Compact Dry VP method for screening raw seafood for total Vibrio parahaemolyticus.

Compact Dry VP (CDVP) is a ready-to-use method for enumerating Vibrio parahaemolyticus in food. The presterilized plates contain a culture medium comprising peptone, NaCl, bile salts, antibiotics, chromogenic substrates, and polysaccharide gum as a cold water-soluble gelling. After diluting raw seafood samples in a phosphate-buffered saline solution, a 1-ml aliquot was inoculated onto the center of the plate and allowed to diffuse by capillary action. Blue-green colonies forming on the plates were counted after 18 to 20 h of incubation at 35 degrees C. A total of 85 V. parahaemolyticus strains (62 tdh+ strains and 23 tdh- strains) were studied for inclusivity, 81 (95.3 %) of which produced blue-green colonies. When 97 strains (14 strains of Vibrio spp., 33 strains of coliform bacteria, and 50 strains of noncoliform bacteria) were assessed for exclusivity, 10 strains of Vibrio spp. produced non-blue-green colonies, and 87 strains failed to grow. The CDVP and U.S. Food and Drug Administration Bacteriological Analytical Manual (FDA-BAM) methods were compared with the use of four different types of raw seafood that were inoculated with four different V. parahaemolyticus strains. For raw tuna and oysters, the FDA-BAM colony lift method was used, whereas the FDA-BAM most-probable-number method was used for salmon and scallop. The linear correlation coefficients between the CDVP and FDA-BAM methods were 0.99 for fresh raw tuna, 0.95 for fresh raw oysters, 0.95 for frozen raw salmon, and 0.95 for frozen raw scallops. These results suggest that the CDVP method is useful for screening raw seafood for V. parahaemolyticus.

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."

Comparison of the compact dry CF with the most probable number method (AOAC official method 966.24) for enumeration of coliform bacteria in raw meats. Performance-Tested Methods" 110401.

Compact Dry CF is a ready-to-use test method for the enumeration of 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 into 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 CF medium plates were validated with 5 different raw meats. The performance tests were conducted at 35 degrees C. In all studies performed, no apparent differences were observed between the Compact Dry CF method and the AOAC Official Method 966.24 results. For the accuracy claim (n = 75), a correlation factor of r2 = 0.91 (coliform) could be assigned, as stated in the application for Performance-Tested Method. No significant variations in coliform bacterial counts were observed with different production lots or plates of diverse storage age by the quality consistency and storage robustness studies.

Comparison of the compact dry YM with the FDA BAM method for enumeration of yeasts and molds in fruit-based products. Performance-Tested Method 100401.

Compact Dry YM is a ready-to-use test method for the enumeration of yeasts and molds in fruit-based products. The plates are presterilized and contain nonwoven incorporated nutrients supplemented with antibiotics and a chromogenic enzyme substrate, 5-bromo-4-chloro-3-indoxyl phosphate, p-toluidine salt (X-phos), to facilitate counting dyes, and a cold water-soluble gelling agent. The medium should be rehydrated with 1 mL of diluted sample inoculated onto the center of the self-diffusible medium, allowing the solution to diffuse immediately by capillary action. The plate can be incubated at 25 degrees C for 7 days. The Compact Dry YM medium plates were validated as an analysis tool determining colony forming units of yeasts and molds from a variety of fruit-based products using 5 different fruit products. The performance tests were conducted at 25 degrees C. In all studies performed, no apparent differences were observed between the Compact Dry YM method and the U.S. Food and Drug Administration's Bacteriological Analytical Manual (FDA BAM) method results (P > 0.05). For the accuracy claim (n = 75), a correlation factor of r2 = 0.99 could be assigned, as stated in the application for Performance-Tested Method.

Comparison of the compact dry TC method with the standard pour plate method (AOAC official method 966.23) for determining aerobic colony counts in food samples: Performance-tested method.

Compact Dry TC qualifies as a rapid method kit for determining aerobic colony counts in foods. The plates are presterilized and contain culture medium and a cold-soluble gelling agent. The medium is rehydrated by inoculating 1 mL diluted sample into the center of the self-diffusible medium and allowing the solution to diffuse by capillary action. The plates can then be incubated and the colonies counted without any additional steps. The Compact Dry TC method was validated with 5 different raw meats. The performance tests were conducted at 35 degrees and 30 degrees C. In all required performance studies, no apparent differences were observed between the Compact Dry TC method and the Standard Pour Plate method (AOAC Official Method 966.23) for the detection level of aerobic microorganisms. For the accuracy claim (n = 60), a correlation factor of r2(35) = 0.9977 (35 degrees C) and r2(30) = 0.9932 (30 degrees C) could be assigned, as stated in the application for "Performance Tested Method." Quality consistency and storage robustness studies, showed no significant variations in plate count results with different production lots or plates of diverse storage age.

Comparative evaluation of the Desoxycholate Agar Nissui Food Stamp and swab methods for estimating coliform organisms in poultry processing plants after cleaning in Japan.

Bacterial control in poultry processing plants is very important, but the swab method for estimating bacterial contamination is somewhat troublesome in routine work. We compared the Desoxycholate Agar Nissui Food Stamp (DA-NFS) based on the agar contact method with the swab method to estimate coliform organisms from various equipments in four poultry processing plants after cleaning. Overall 104 surfaces for coliform organisms were evaluated. The results from 98 (94.2%) surfaces for coliform organisms were equivalent by the DA-NFS and swab methods and there were no significant differences between two methods (P > 0.05). The correlation coefficient between the DA-NFS and swab methods was 0.91. We conclude that the DA-NFS could be useful for routine coliform organisms examination in poultry processing plants after cleaning in Japan.

Evaluation of a new agar medium containing cetrimide, kanamycin and nalidixic acid for isolation and enhancement of pigment production of Pseudomonas aeruginosa in clinical samples.

A new selective agar medium (CKNA), containing cetrimide 0.3 g/l, kanamycin sulfate 50 mg/l, and nalidixic acid 5 mg/l, was developed for the isolation of Pseudomonas aeruginosa. It was compared to Nalidixic Acid Cetrimide agar (NAC), Pseudomonas Aeruginosa Selective Agar (PASA) and Pseudosel(TM) agar (CET) using 1,148 clinical specimens. The sensitivities rates of P. aeruginosa with CKNA, NAC, PASA, and CET were 88.2%, 81.3%, 79.2%, and 84.0%, respectively. The specificities of CKNA, NAC, PASA, and CET were 99.2%, 98.4%, 99.2%, and 99.7%, respectively.