Evaluation of risk based microbiological criteria for Campylobacter in broiler carcasses in Belgium using TRiMiCri

Campylobacteriosis is the most frequently reported foodborne zoonosis worldwide. Consumer´s exposure to Campylobacter might be reduced by establishing a microbiological criterion (MC) for Campylobacter on broiler meat. In the present study two possible approaches were evaluated, using the freely available software tool for risk based microbiological criteria TRiMiCri (http://tools.food.dtu.dk/trimicri). The first approach was the traditional one that implies a microbiological limit (ML-MC) and the second one which is based on the relative risk estimate (RRL-MC). The analyses were based on Campylobacter quantitative data collected from 28 Campylobacter positive bathes processed in 6 Belgian broiler slaughterhouses. To evaluate the performance of ML-MC, n=6, different c (0,1,2) and m (100,1 000,10 000) were used. Results showed that more than 90% of Campylobacter positive batches were not complying with strict ML criteria based on the m=100 for all applied combination of c. The RRL approach requires a baseline risk which was estimated based on the Campylobacter baseline data collected in Belgium in 2008. Approximately 60% of evaluated Campylobacter positive batches account for higher risk than the baseline risk. For both approaches, application of less stringent criteria results in lower percentage of NC and higher minimum relative residual risks (MRRR; it refers to the change in risk when all batches are sampled and all NC batches undergo treatment that effectively eliminates Campylobacter so they are replaced by zero risk batches). It was also observed that the number of samples (n) had little effect on risk estimates. Additionally, the results from ML-MC and RRL-MC follow the same curve when plotting percentage of NC against MRRR. However, for RRL-MC the percentage of NC batches and MRRR was lower and higher, respectively. To conclude, obtained results indicate that TRiMiCri is a useful and user friendly tool to make a risk based decision on the choice of the MC

Structural Basis of GLUT1 Inhibition by Cytoplasmic ATP

Cytoplasmic ATP inhibits human erythrocyte glucose transport protein (GLUT1)–mediated glucose transport in human red blood cells by reducing net glucose transport but not exchange glucose transport (Cloherty, E.K., D.L. Diamond, K.S. Heard, and A. Carruthers. 1996. Biochemistry. 35:13231–13239). We investigated the mechanism of ATP regulation of GLUT1 by identifying GLUT1 domains that undergo significant conformational change upon GLUT1–ATP interaction. ATP (but not GTP) protects GLUT1 against tryptic digestion. Immunoblot analysis indicates that ATP protection extends across multiple GLUT1 domains. Peptide-directed antibody binding to full-length GLUT1 is reduced by ATP at two specific locations: exofacial loop 7–8 and the cytoplasmic C terminus. C-terminal antibody binding to wild-type GLUT1 expressed in HEK cells is inhibited by ATP but binding of the same antibody to a GLUT1–GLUT4 chimera in which loop 6–7 of GLUT1 is substituted with loop 6–7 of GLUT4 is unaffected. ATP reduces GLUT1 lysine covalent modification by sulfo-NHS-LC-biotin by 40%. AMP is without effect on lysine accessibility but antagonizes ATP inhibition of lysine modification. Tandem electrospray ionization mass spectrometry analysis indicates that ATP reduces covalent modification of lysine residues 245, 255, 256, and 477, whereas labeling at lysine residues 225, 229, and 230 is unchanged. Exogenous, intracellular GLUT1 C-terminal peptide mimics ATP modulation of transport whereas C-terminal peptide-directed IgGs inhibit ATP modulation of glucose transport. These findings suggest that transport regulation involves ATP-dependent conformational changes in (or interactions between) the GLUT1 C terminus and the C-terminal half of GLUT1 cytoplasmic loop 6–7

Exposure assessment of foodborne pathogens in pork in Belgium

The aim of this study was to assess the exposure of the most incident foodborne pathogens in the Belgian meat production chain. The prevalence of Salmonella, Campylobacter and Listeria monocytogenes were evaluated in carcasses (swabs), retail cuts, minced meat and meat products of pork. The investigation was made each year since 1997, using official methods from the Ministry of Public Health for Salmonella and Campylobacter and the Vidas Listeria monocytogenes method. More than 10 % of each matrix were contaminated with Salmonella. For minced meat and meat products, the contamination rate were respectively round 20 % and 3 – 6 % for Listeria monocytogenes. Under 5 % of minced meat samples were positive for Campylobacter. For minced meat, the contamination has also been assessed according to the location of sampling (agreed, low capacity establishments or retail level). The characterisation of bacterial species allows the comparison between meat and human isolates

Virulence profiling and quantification of verocytotoxin-producing Escherichia coli O145:H28 and O26:H11 isolated during an ice cream-related hemolytic uremic syndrome outbreak

In September-October 2007, a mixed-serotype outbreak of verocytotoxin-producing Escherichia coli (VTEC) O145:H28 and O26:H11 occurred in the province of Antwerp, Belgium. Five girls aged between 2 and 11 years developed hemolytic uremic syndrome, and seven other coexposed persons with bloody diarrhea were identified. Laboratory confirmation of O145:H28 infection was obtained for three hemolytic uremic syndrome patients, one of whom was coinfected with O26:H11. The epidemiological and laboratory investigations revealed ice cream as the most likely source of the outbreak. The ice cream was produced at a local dairy farm using pasteurized milk. VTEC of both serotypes with indistinguishable pulsed-field gel electrophoresis patterns were isolated from patients, ice cream, and environmental samples. Quantitative analysis of the ice cream indicated concentrations of 2.4 and 0.03 CFU/g for VTEC O145 and O26, respectively. Virulence typing revealed that the repertoire of virulence genes carried by the O145:H28 outbreak strain was comparable to that of O157 VTEC and more exhaustive as compared to the O26:H11 outbreak strain and nonrelated clinical strains belonging to these serotypes. Taken together, these data suggest that O145:H28 played the most important role in this outbreak