IAFP 2009 Program Book and Abstract Book - AB114
sorbitol MacConkey agar (SMAC). After incubating at 36°C for 24 h, five colonies per each sample from the SMAC were reanalyzed, using the multiplex real-time PCR. The positive colonies in the real-time PCR were streaked on Tryptic Soy Agar with 0.6% yeast extract (TSAYE) and incubated at 36°C for 24 h, followed by biochemical confirmation testing using API 20 E. Results: In the presumptive screening of enriched mEC, all samples from omasum and large intestine were positive for at least one of the two toxin genes (stx1 and stx2) by the multiplex real-time PCR method, while 88% (30/34) were positive in abomasums. When five presumptive colonies per samples from the SMAC were reanalyzed using real-time PCR and biochemical confirmation, total STEC prevalence (STEC sample number/Total sample number) rates were 25% (8/32) in omasum, 20% (7/34) in abomasum, 37.5% (6/16) in large intestine and a total of 32 STEC strains were isolated from all samples (11 strains from omasum, 10 strains from abomasum and 11 strains from large intestine). Significance: This study shows that a high percentage of the edible by-products from cattle were contaminated with STEC, and the multiplex PCR can be used as a confirmatory means of isolating STEC. P2-28 Reclassification of ATCC® 49444™ from Staphylococcus aureus to Staphyloccus pseudintermedius KIMBERLY J. RAMSEY, Erin C. Cleveland, Marian L. McKee and Brian J. Beck ATCC, Bacteriology, Manassas, VA, USA Introduction: Strain ATCC 49444, currently known as Staphylococcus aureus, has been designated as a component strain in both FDA’s Bacteriological Analytical Manual and USDA-FSIS’s Laboratory Guidebook (MLG 8.06) for the CAMP (Christie-Atkins-Munch-Peterson) test for speciation of Listeria isolates. This assay is based on the synergistic hemolytic activity of L. monocytogenes that occurs in the presence of beta-lysin producing staphylococci. Purpose: While conducting authentication tests on this strain, we observed that its genetic and biochemical characteristics were not consistent with its designation as S. aureus, but rather S. intermedius or the recently described S. pseudintermedius, depending on the method used for identification. Methods: A culture from the original seed lot was subjected to a series of polyphasic tests and compared to closely related Staphylococcal Type cultures as well as commonly used QC strains from the ATCC collection to determine the correct identification of this strain. Results: Phenotypic analysis of ATCC 49444 using a bioMérieux Vitek 2 Compact identified the strain as S. intermedius. Since S. intermedius and S. pseudintermedius strains cannot be distinguished biochemically, the nucleotide sequences of the 16S rDNA and sodA genes from ATCC 49444 were determined and found to be consistent with other S. pseudintermedius strains. Riboprint analysis of ATCC 49444 indicated a low (0.1 to 0.3) similarity index with S. aureus or S. intermedius strains, but > 0.75 similarity index with S. pseudintermedius isolates. A functional comparison of ATCC 49444 with S. aureus ATCC 25923 utilizing the CAMP test demonstrated equivalent hemolytic enhancement with L. monocytogenes. Significance: While this study demonstrates that ATCC 49444 is a valid component strain for the CAMP test, it is proposed that ATCC 49444 be reclassified as S. pseudintermedius and that users worldwide be alerted to this name change. P2-29 Evaluation of the TEMPO® STA Method for the Enumeration of Staphylococcus aureus in Foods JOHN C. MILLS, Judith Colón-Reveles, Ronald Johnson and Gregory Devulder bioMérieux, Inc., Hazelwood, MO, USA Introduction: The TEMPO® STA method was developed for the automated enumeration of Staphylococcus aureus in foods. The method utilizes a selective dehydrated culture medium and an enumeration card containing 48 wells across 3 different dilutions for the automatic determination of the Most Probable Number. Purpose: As part of the AOAC Research Institute validation process, the alternative method was compared to the FDA Bacteriological Analytical Manual (BAM) method for all foods. Methods: Fourteen naturally and artificially contaminated foods were tested, including meat, poultry, dairy, seafood, confectionary, and vegetables. Five replicates of three lots for each food were tested, for a total of 210 samples. A 1:10 dilution of each sample was prepared and stomached for 2 min. For each diluted and stomached sample, 1.0 ml of diluted food sample was added to a TEMPO medium vial that had been reconstituted with 3.0 ml of sterile distilled water. The inoculated medium in the vial was then transferred and sealed into the STA card by the automated TEMPO filler. The inoculated cards were incubated for 26 h at 35 ± 1°C. Cards were read using the automated TEMPO reader. Standard method testing was performed as detailed in the BAM. Results: For the majority of samples tested, there was no significant difference for both the mean log counts and repeatability between the alternative method and the standard method, using a paired t-test and f-test at the 5% level. Significance: TEMPO STA provides an automated, accurate method for the enumeration of Staphylococcus aureus in foods. In addition, the TEMPO STA offers a considerable time and labor savings when compared to the reference methods tested in this study. P2-30 Comparison of the 3M™ Tecra™ Staphylococcus aureus Visual Immunoassay to the United States Pharmacopeia Standard Method for the Detection of Staphylococcus aureus in Raw and Finished Dietary Supplements CHRISTINE A. BINSFELD and Enrique Morales 3M, Food Safety Dept., St. Paul, MN, USA Introduction: Staphylococcus aureus has been reported to be a microbial contaminant in many foods, cosmetic/pharmaceutical products and dietary supplements. Because of the pathogenicity of S. aureus, the United States Pharmacopeia (USP) requires its absence in certain non-sterile pharmaceutical raw materials and finished products. There is a need to apply new technologies to conduct the microbiological monitoring of raw and finished products. These technologies need to be rapid, sensitive, accurate, cost effective, and easy to use. The standard microbiological methods require 4 to 5 days for the complete isolation and identification of S. aureus. 114 — Abstract Book Poster
IAFP 2009 Program Book and Abstract Book
Table of Contents for the Digital Edition of IAFP 2009 Program Book and Abstract Book
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