Comparison of dkgB-linked intergenic sequence ribotyping to DNA microarray hybridization for assigning serotype to Salmonella enterica.

Two DNA-based methods were compared for the ability to assign serotype to 139 isolates of Salmonella enterica ssp. I. Intergenic sequence ribotyping (ISR) evaluated single nucleotide polymorphisms occurring in a 5S ribosomal gene region and flanking sequences bordering the gene dkgB. A DNA microarray hybridization method that assessed the presence and the absence of sets of genes was the second method. Serotype was assigned for 128 (92.1%) of submissions by the two DNA methods. ISR detected mixtures of serotypes within single colonies and it cost substantially less than Kauffmann-White serotyping and DNA microarray hybridization. Decreasing the cost of serotyping S. enterica while maintaining reliability may encourage routine testing and research.

Selection and characterization of cellulose-deficient derivates of shiga toxin-producing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) is known to have several defense mechanisms, one of which is the production of extracellular substances including cellulose. The goal of this study was to prepare pairs of STEC cultures for use in future studies designed to address the role of cellulose in protecting the cells of STEC for survival under adverse environmental conditions. Cells of STEC deficient in cellulose production were separated from cellulose-proficient wild-type cells. The identities of the two types of cells were confirmed using serotyping and pulsed-field gel electrophoresis (PFGE). Selected growth characteristics of the two types of cells were determined using three phenotype microarray plates, PM9, PM10, and PM11. The cellulose-deficient and cellulose-proficient cells in each STEC pair shared the same serotype and PFGE profile. The deficiency in cellulose production did not significantly (P > 0.05) affect the growth characteristics of STEC cells under 191 of the 210 tested growth conditions. Significant differences in growth between the two types of cells were observed only in the presence of two antibiotics, a short chain fatty acid, and high concentrations of osmolytes, as well as under extreme acidic and alkaline pH. These results suggest that deficiency in cellulose production did not alter the serological property, PFGE profile, and growth characteristics of selected STEC strains under optimal growth conditions. The STEC strains and their cellulose-deficient derivates could be useful for studying the role of cellulose in protecting the cells of STEC for survival under adverse environmental conditions.

Behavior of Bacillus anthracis strains Sterne and Ames K0610 in sterile raw ground beef.

The behavior of Bacillus anthracis Sterne spores in sterile raw ground beef was measured at storage temperatures of 2 to 70 degrees C, encompassing both bacterial growth and death. B. anthracis Sterne was weakly inactivated (-0.003 to -0.014 log10 CFU/h) at storage temperatures of 2 to 16 degrees C and at temperatures greater than and equal to 45 degrees C. Growth was observed from 17 to 44 degrees C. At these intermediate temperatures, B. anthracis Sterne displayed growth patterns with lag, growth, and stationary phases. The lag phase duration decreased with increasing temperature and ranged from approximately 3 to 53 h. The growth rate increased with increasing temperature from 0.011 to 0.496 log10 CFU/h. Maximum population densities (MPDs) ranged from 5.9 to 7.9 log10 CFU/g. In addition, the fate of B. anthracis Ames K0610 was measured at 10, 15, 25, 30, 35, 40, and 70 degrees C to compare its behavior with that of Sterne. There were no significant differences between the Ames and Sterne strains for both growth rate and lag time. However, the Ames strain displayed an MPD that was 1.0 to 1.6 times higher than that of the Sterne strain at 30, 35, and 40 degrees C. Ames K0610 spores were rapidly inactivated at temperatures greater than or equal to 45 degrees C. The inability of B. anthracis to grow between 2 and 16 degrees C, a relatively low growth rate, and inactivation at elevated temperatures would likely reduce the risk for recommended ground-beef handling and preparation procedures.