Occurrence and Levels of Salmonella, Enterohemorrhagic Escherichia coli, and Listeria in Raw Wheat.

HIGHLIGHTS: Prevalence of Salmonella and E. coli in raw wheat emphasizes the need to cook wheat products. 3,891 grain samples were tested for E. coli and Salmonella; 1,285 were tested for Listeria. Of wheat berries sampled, 0.44% were positive for E. coli and 1.23% were positive for Salmonella. Salmonella diversity was high, indicating various animal sources that are difficult to prevent. Cooking wheat products is the best preventative measure against foodborne illness from wheat.

Microbial successions are associated with changes in chemical profiles of a model refrigerated fresh pork sausage during an 80-day shelf life study.

Fresh pork sausage is produced without a microbial kill step and therefore chilled or frozen to control microbial growth. In this report, the microbiota in a chilled fresh pork sausage model produced with or without an antimicrobial combination of sodium lactate and sodium diacetate was studied using a combination of traditional microbiological methods and deep pyrosequencing of 16S rRNA gene amplicons. In the untreated system, microbial populations rose from 10(2) to 10(6) CFU/g within 15 days of storage at 4°C, peaking at nearly 10(8) CFU/g by day 30. Pyrosequencing revealed a complex community at day 0, with taxa belonging to the Bacilli, Gammaproteobacteria, Betaproteobacteria, Actinobacteria, Bacteroidetes, and Clostridia. During storage at 4°C, the untreated system displayed a complex succession, with species of Weissella and Leuconostoc that dominate the product at day 0 being displaced by species of Pseudomonas (P. lini and P. psychrophila) within 15 days. By day 30, a second wave of taxa (Lactobacillus graminis, Carnobacterium divergens, Buttiauxella brennerae, Yersinia mollaretti, and a taxon of Serratia) dominated the population, and this succession coincided with significant chemical changes in the matrix. Treatment with lactate-diacetate altered the dynamics dramatically, yielding a monophasic growth curve of a single species of Lactobacillus (L. graminis), followed by a uniform selective die-off of the majority of species in the population. Of the six species of Lactobacillus that were routinely detected, L. graminis became the dominant member in all samples, and its origins were traced to the spice blend used in the formulation.

Influence of organic buffers on bacteriocin production by Streptococcus thermophilus ST110.

The effect of the organic buffer salts MES, MOPS, and PIPES on the growth of S. thermophilus ST110, medium pH, and accumulation of the antipediococcal bacteriocin thermophilin 110 were evaluated in whey permeate media over a period of 24 h. In nonbuffered medium, thermophilin 110 production at 37 degrees C paralleled the growth of S. thermophilus ST110 and reached a maximum after 8-10 h. Addition of organic buffer salts decreased the drop in medium pH and resulted in increased biomass (dry cells; microg/mL) and higher yields of thermophilin 110 (units/microg cells). The best results were obtained by the addition of 1% (w/v) MES to the medium, which reduced the pH drop to 1.8 units after 10 h of growth (compared to 2.3 pH units in the control) and resulted in a 1.5-fold increase in cell mass (495 microg/mL) and a 7-fold increase in thermophilin 110 yield (77 units/microg dry cells) over the control. The results showed that whey permeate-based media may be suitable for producing large amounts of thermophilin 110 needed for controlling spoilage pediococci in industrial wine and beer fermentations.

Use of octanoic acid as a postlethality treatment to reduce Listeria monocytogenes on ready-to-eat meat and poultry products.

The antilisterial efficacy and organoleptic impact of an octanoic acid (OA)-based treatment for ready-to-eat (RTE) meat and poultry products were investigated. Whole-muscle and comminuted RTE products were inoculated with a five-strain mixture of Listeria monocytogenes. The OA treatments were applied to the surface of RTE products by dispensing a specific volume of solution directly into the final package prior to vacuum sealing. Once sealed, the vacuum-packaged RTE products containing OA were immersed in water heated to 93.3 degrees C (200 degrees F) for 2 s to effect adequate film shrinkage. Extending the time at which the packaged, treated RTE products were exposed to water heated to 93.3 degrees C was also evaluated with a commercial cascading shrink tunnel fitted with a modified drip pan. Once treated, RTE products were examined for survivor populations of L. monocytogenes after 24 h of storage at 5 degrees C. Sensory evaluation was conducted with a 60-member trained panel on 11 uninoculated, treated RTE products. The OA treatment of RTE products reduced L. monocytogenes numbers to between 0.85 log CFU per sample (oil-browned turkey) and 2.89 log CFU per sample (cured ham) when compared with controls. The antilisterial activity of OA was improved by increasing the duration of the heat shrink exposure. Specifically, reductions of L. monocytogenes ranged from 1.46 log CFU per sample (oil-browned turkey) to 3.34 log CFU per sample (cured ham). Results from the sensory evaluation demonstrated that 10 of the 11 treated RTE products were not perceived as different (P < or = 0.05) from the untreated controls. Panelists detected reduced (P < or = 0.05) smoke flavor intensity with treated mesquite turkey, although the treated product was viewed as acceptable. Results demonstrate the effectiveness of OA as a postlethality treatment meeting U.S. Food Safety and Inspection Service regulatory guidelines for RTE meat and poultry products with minimal impact on sensory quality.

Relatedness of Listeria monocytogenes Isolates recovered from selected ready-to-eat foods and listeriosis patients in the United States.

Pulsed-field gel electrophoresis and serotyping were performed for 544 isolates of Listeria monocytogenes, including 502 isolates recovered from contaminated samples from 31,705 retail ready-to-eat (RTE) food products and 42 isolates recovered from human cases of listeriosis. The isolates were from Maryland (294 isolates) and California (250 isolates) and were collected in 2000 and 2001. The isolates were placed into 16 AscI pulsogroups (level of relatedness within each group, > or =66%), 139 AscI pulsotypes (levels of relatedness, > or =25% to 100%), and eight serotypes (serotypes 1/2a, 1/2b, 1/2c, 3a, 3b, 4b, 4c, and 4d). The most frequently found pulsotypes belonged to either pulsogroup A (150 food isolates plus 4 clinical isolates) or pulsogroup B (104 food isolates plus 5 clinical isolates). The majority of the 502 food isolates were either serotype 1/2a (298 isolates) or serotype 1/2b (133 isolates), whereas the majority of the 42 clinical isolates were either serotype 1/2a (19 isolates) or serotype 4b (15 isolates). Additionally, 13 clinical isolates displayed pulsotypes also found in food isolates, whereas the remaining 29 clinical isolates displayed 24 unique pulsotypes. These data indicate that most (86%) of the L. monocytogenes subtypes found in the RTE foods sampled belonged to only two serotypes and that 90% of the isolates displayed 73 pulsotypes, with 107 isolates displaying pulsotype 1. These data should help define the distribution and relatedness of isolates found in RTE foods in comparison with isolates that cause listeriosis.

Thermophilin 110: a bacteriocin of Streptococcus thermophilus ST110.

A screen of thermophilic lactic acid bacteria identified Streptococcus thermophilus strain ST110 as the putative producer of a bacteriocin with high level of activity against pediococci. Thermophilin 110 was isolated from culture supernatant after 16 h of growth and partially purified by a chloroform extraction procedure. The bacteriocin inhibited the growth of several lactic acid bacteria and in the case of Pediococcus acidilactici, it induced cell lysis with the concomitant release of OD260-absorbing material and intracellular enzymes. SDS-PAGE analysis revealed two components with estimated sizes between 4.0 kDa and 4.5 kDa, respectively, with possible involvement in bacteriocin activity as indicated by agar overlay assays with P. acidilactici as the target organism. Thermophilin 110 was inactivated by several proteolytic enzymes and also by alpha-amylase, which indicated the putative requirement of a glycosidic component for activity. The bacteriocin produced by S. thermophilus may be especially useful in the food processing industries to control spoilage caused by pediococci.

Catabolite repression and virulence gene expression in Listeria monocytogenes.

Previous studies have suggested that carbohydrates may affect expression of virulence genes in Listeria monocytogenes. Which carbohydrates influence virulence gene expression and how carbohydrates mediate expression, however, is not clear. The goal of this work was to examine how carbohydrates affect virulence gene expression in L. monocytogenes 10403S. Growth studies were conducted in medium containing glucose and various sugars. Metabolism of arbutin, arabitol, cellobiose, mannose, maltose, trehalose, and salicin were repressed in the presence of glucose. Only when glucose was consumed were these sugars fermented, indicating that catabolite repression by glucose had occurred. To determine whether virulence gene expression was also influenced by catabolite repression, we performed primer extension experiments, using primers for hly and prfA, which encode for a hemolysin and the regulator protein PrfA, respectively. In the presence of cellobiose and arbutin, transcription of hemolysin was reduced. However, none of the sugars affected transcription of prfA. The results demonstrate that catabolite repression occurs in L. monocytogenes and suggests that, at least in strain 10403S, cellobiose and arbutin repress expression of hemolysin.