Declan Bolton

Food safety knowledge of consumers and the microbiological and temperature status of their refrigerators.

The objectives of this study were to examine domestic food safety knowledge levels of consumers, establish the levels and incidence of bacterial contamination and operational temperatures in domestic refrigerators, and identify areas in which consumer food safety education is necessary in Ireland. A food safety knowledge questionnaire applied to a representative sample of households (n = 1,020) throughout the island of Ireland found the gaps in consumer food safety knowledge. Analysis of swab samples (n = 900) recovered from the domestic refrigerators in these households showed average total viable counts of 7.1 log CFU/cm2 and average total coliform counts of 4.0 log CFU/cm2. Analysis of swab samples also detected the incidence of Staphylococcus aureus (41%), Escherichia coli (6%), Salmonella enterica (7%), Listeria monocytogenes (6%), and Yersinia enterocolitica (2%). Campylobacter jejuni and E. coli O157:H7 were not detected in domestic refrigerators. The temperature profiles of a subset of the sampled refrigerators (100) were monitored for 72 h, and 59% were found to operate, on average, at temperatures above the recommended 5 degrees C. Knowledge and temperature survey results varied considerably, but consumers who scored better in terms of basic food safety knowledge had reduced levels of bacterial contamination in their refrigerators and reported a reduced incidence of food-associated illnesses. This study confirms the effect of basic food hygiene knowledge on hygienic practice and identifies specific areasfor emphasis in the development and delivery of effective food safety risk communication messages to consumers.

The‐survival‐characteristics‐of‐a‐non‐toxigenic‐strain‐of‐Escherichia‐coli‐O157:H7

The‐survival‐characteristics‐of‐a‐non‐toxigenic,‐antibiotic‐resistant‐strain‐of‐Escherichia‐coli‐O157:H7‐in‐bovine‐faeces‐were‐investigated.‐Faecal‐samples‐were‐inoculated‐with 108‐9‐cfu‐g−1‐of‐the‐organism‐and‐(i)‐stored‐in‐closed‐plastic‐containers‐at‐10 °C, (ii) stored in closed plastic containers placed outside or (iii) decanted onto the surface of grazing land. Recovery and enumeration on Sorbitol MacConkey Agar (SMAC) and Tryptic Soya Agar (TSA) revealed that the E. coli O157:H7 numbers in both enclosed samples (i and ii) had decreased by 4·5–5·5 log10 cfu g−1 within 99 d. Numbers in samples decanted onto grassland (iii) decreased by 4·0–5·0 log10 cfu g−1 within 50 d but the organism was still detectable in the surrounding soil for up to 99 d. Persistence of E. coli O157:H7 in bovine faeces and contaminated pastures may therefore be an important factor in the initial infection and re‐infection of cattle.

Washing and chilling as critical control points in pork slaughter hazard analysis and critical control point (HACCP) systems

Aims: The aim of this research was to examine the effects of preslaughter washing, pre‐evisceration washing, final carcass washing and chilling on final carcass quality and to evaluate these operations as possible critical control points (CCPs) within a pork slaughter hazard analysis and critical control point (HACCP) system.

Studies to determine the critical control points in pork slaughter hazard analysis and critical control point systems.

Aerobic mesophilic counts (AMC), coliform (CC) and coliform resuscitation counts (CRCs) were obtained by swabbing 50 cm(2) areas at three sites (ham, belly and neck) on pig carcasses, after each of seven stages of the slaughter/dressing process (bleeding, scalding, dehairing, singeing, polishing, evisceration and chilling). In most cases, there were no statistical differences (P>0.05) among the counts derived by these three methods. Reductions in counts at individual sites were observed after scalding (3.5 log(10) cfu cm(-2)), and singeing (2.5 log(10) cfu cm(-2)). Increases in counts at individual sites were observed after dehairing (2.0 log(10) cfu cm(-2)) and polishing (1.5 log(10) cfu cm(-2)). The incidence of Salmonella on pig carcasses was also obtained by swabbing the outside surfaces of 100 half carcasses. Information on the incidence of Salmonella in scald tank water (108 samples) was also investigated. Carcass swabs and scald tank water were examined for the presence of Salmonella using standard enrichment methods. Salmonella were detected on 31% of carcasses immediately after bleeding, 7% of carcasses immediately after dehairing and evisceration, and 1% of carcasses immediately after scalding. Serovars included Salmonella Typhimurium, Salmonella Hadar, Salmonella Infantis and Salmonella Derby. No Salmonella were recovered from samples of scald tank water. The impact of pig slaughter/dressing processes on carcass microbiology and their potential use as critical control points (CCPs) during pork production are discussed.

Characterisation and transferability of antibiotic resistance genes from lactic acid bacteria isolated from Irish pork and beef abattoirs.

Lactic acid bacteria isolated from Irish pork and beef abattoirs were analysed for their susceptibility to antimicrobials. Thirty-seven isolates (12 enterococci, 10 lactobacilli, 8 streptococci, 3 lactococci, 2 Leuconostoc, and 2 pediococci) were examined for phenotypic resistance using the E-test and their minimum inhibitory concentration to a panel of six antibiotics (ampicillin, chloramphenicol, erythromycin, streptomycin, tetracycline, and vancomycin) was recorded. The corresponding genetic determinants responsible were characterised by PCR. Also, the transferability of these resistance markers was assessed in filter mating assays. Of the 37 isolates, 33 were found to be resistant to one or more antibiotics. All strains were susceptible to ampicillin and chloramphenicol. The erm(B) and msrA/B genes were detected among the 11 erythromycin-resistant strains of enterococci, lactobacilli, and streptococci. Two tetracycline-resistant strains, Lactobacillus plantarum and Leuconostoc mesenteroides spp., contained tet(M) and tet(S) genes respectively. Intrinsic streptomycin resistance was observed in lactobacilli, streptococci, lactococci and Leuconostoc species; none of the common genetic determinants (strA, strB, aadA, aadE) were identified. Four of 10 strains of Enterococcus faecium were resistant to vancomycin; however, no corresponding genetic determinants for this phenotype were identified. Enterococcus faecalis strains were susceptible to vancomycin. L. plantarum, L. mesenteroides and Pediococcus pentosaceus were intrinsically resistant to vancomycin. Transfer of antibiotic resistance determinants was demonstrated in one strain, wherein the tet(M) gene of L. plantarum (23) isolated from a pork abattoir was transferred to Lactococcus lactis BU-2-60 and to E. faecalis JH2-2. This study identified the presence of antibiotic resistance markers in Irish meat isolates and, in one example, resistance was conjugally transferred to other LAB strains.

The effects of preslaughter washing on the reduction of Escherichia coli O157:H7 transfer from cattle hides to carcasses during slaughter

C.M. BYRNE, D.J. BOLTON, J.J. SHERIDAN, D.A. MCDOWELL and I.S. BLAIR.2000. Fresh bovine faeces were inoculated with a non‐toxigenic, antibiotic resistant strain of Escherichia coli O157:H7, spread on the rump areas of 30 heifers and allowed to dry for 24 h. Ten of the cattle then entered the normal slaughter process without further treatment. The remaining cattle were washed with a powerhose for 1 min (10 animals) and 3 min (10 animals) before entering the normal slaughter process. Both washing treatments removed all visible faecal materials on the live animals although a significant reduction (P < 0·05) in E. coli O157:H7 levels on the hides was only observed on those animals which were powerhosed for 3 min. After slaughter, E. coli O157:H7 was detected on carcasses and on the knives and hands of operatives. Preslaughter washing for 3 min did not statistically reduce the numbers of E. coli O157:H7 transferred from the hide to the carcass during slaughter. However, the organism was not detected on three of the four areas of the carcass sampled, indicating that washing may be a suitable method of decontamination animal hides before slaughter and as such deserves further investigation.

Campylobacter virulence and survival factors

Despite over 30 years of research, campylobacteriosis is the most prevalent foodborne bacterial infection in many countries including in the European Union and the United States of America. However, relatively little is known about the virulence factors in Campylobacter or how an apparently fragile organism can survive in the food chain, often with enhanced pathogenicity. This review collates information on the virulence and survival determinants including motility, chemotaxis, adhesion, invasion, multidrug resistance, bile resistance and stress response factors. It discusses their function in transition through the food processing environment and human infection. In doing so it provides a fundamental understanding of Campylobacter, critical for improved diagnosis, surveillance and control.

Verocytotoxigenic (Shiga Toxin–Producing) Escherichia coli: Virulence Factors and Pathogenicity in the Farm to Fork Paradigm

Verocytotoxigenic Escherichia coli (VTEC) are a good example of the evolution and emergence of pathogenic E. coli. Unknown before the late 1970s, these bacteria are a major cause of hemorrhagic colitis and hemolytic uremic syndrome worldwide. The production of verocytotoxins is the main virulence feature of VTEC but cannot be solely responsible for full pathogenicity. VTEC associated with severe human disease are usually capable of colonizing the intestinal mucosa with a characteristic attaching-and-effacing mechanism, genetically governed by the locus of enterocyte effacement, and possess other mobile genetic elements carrying additional virulence genes such as plasmids, phages, and pathogenicity islands (e.g., O-I 122). Despite the huge amount of data collected after the sequencing of the full genome of VTEC O157, the virulence and the evolution of the different VTEC serotypes have only been partially unraveled. A greater understanding of the factors governing the development of severe disease in humans and the colonization of animal hosts must be achieved before effective intervention strategies aimed at the reduction of the burden of infection can be developed. Defining all the factors characterizing a fully pathogenic VTEC strain will be crucial to improve the efficacy of the diagnosis of human infections, the surveillance of animal reservoirs, the assessment of public health risks, and the development of control interventions. An overview of the VTEC virulence factors, including their genetic basis and function, would start this process and is the objective of this article.

Serotypes and virulence profiles of non-O157 Shiga toxin-producing Escherichia coli isolates from bovine farms.

ABSTRACT Non-O157 Shiga toxin-producing Escherichia coli (STEC) strains are clinically significant food-borne pathogens. However, there is a dearth of information on serotype prevalence and virulence gene distribution, data essential for the development of public health protection monitoring and control activities for the meat and dairy industries. Thus, the objective of this study was to examine the prevalence of non-O157 STEC on beef and dairy farms and to characterize the isolates in terms of serotype and virulence markers. Bovine fecal samples (n = 1,200) and farm soil samples (n = 600) were collected from 20 farms throughout Ireland over a 12-month period. Shiga toxin-positive samples were cultured and colonies examined for the presence of stx 1 and/or stx 2 genes by PCR. Positive isolates were serotyped and examined for a range of virulence factors, including eaeA, hlyA, tir, espA, espB, katP, espP, etpD, saa, sab, toxB, iha, lpfA O157/OI-141, lpfA O113, and lpfA O157/OI-154. Shiga toxin and intimin genes were further examined for known variants. Significant numbers of fecal (40%) and soil (27%) samples were stx positive, with a surge observed in late summer-early autumn. One hundred seven STEC isolates were recovered, representing 17 serotypes. O26:H11 and O145:H28 were the most clinically significant, with O113:H4 being the most frequently isolated. However, O2:H27, O13/O15:H2, and ONT:H27 also carried stx 1 and/or stx 2 and eaeA and may be emerging pathogens.

Consumer food safety knowledge

Purpose – Consumers have an important role to play in preventing food‐borne disease. The purpose of this study was to demonstrate that consumers could be segmented successfully based on their food safety knowledge and practice.Design/methodology/approach – Principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to data on food safety knowledge and practice, collected by individual face‐to‐face questionnaires with domestic food preparers (n=1,020) and refrigerator swabs (n=726).Findings – From the food safety questionnaires four factors were identified as important for inclusion in the HCA. This analysis identified three groups of consumers based on the knowledge factors; they were “Conscientious” (21 per cent), “Cavalier” (25.4 per cent) and “Careful” (53.3 per cent) food handlers. In terms of food safety knowledge, the higher risk consumers were found to be in the Cavalier food handler group. This group of food handlers also engaged in less hygienic food handling practices...