Comparison of maintenance energy requirement and energetic efficiency between lactating Holstein-Friesian and other groups of dairy cows.

The objectives of the present study were to investigate the effects of cow group on energy expenditure and utilization efficiency. Data used were collated from 32 calorimetric chamber experiments undertaken from 1992 to 2010, with 823 observations from lactating Holstein-Friesian (HF) cows and 112 observations from other groups of lactating cows including Norwegian (n=50), Jersey × HF (n=46), and Norwegian × HF (n=16) cows. The metabolizable energy (ME) requirement for maintenance (MEm) for individual cows was calculated from heat production (HP) minus energy losses from inefficiencies of ME use for lactation, energy retention, and pregnancy. The efficiency of ME use for lactation (kl) was obtained from milk energy output adjusted to zero energy balance (El(0)) divided by ME available for production. The effects of cow groups were first evaluated using Norwegian cows against HF crossbred cows (F1 hybrid, Jersey × HF and Norwegian × HF). The results indicated no significant difference between the 2 groups in terms of energy digestibility, ratio of ME intake over gross energy intake, MEm (MJ per kg of metabolic body weight, MJ/kg(0.75)), or kl. Consequently, their data were combined (categorized as non-HF cows) and used to compare with those of HF cows. Again, we detected no significant difference in energy digestibility, ratio of ME intake over gross energy intake, MEm (MJ/kg(0.75)), or kl between non-HF and HF cows. The effects were further evaluated using linear regression to examine whether any significant differences existed between HF and non-HF cows in terms of relationships between ME intake and energetic parameters. With a common constant, no significant difference was observed between the 2 groups of cows in coefficients in each set of relationships between ME intake (MJ/kg(0.75)) and MEm (MJ/kg(0.75)), El(0) (MJ/kg(0.75)), HP (MJ/kg(0.75)), MEm:ME intake, El(0):ME intake, or HP:ME intake. However, MEm values (MJ/kg(0.75)) were positively related to ME intake (MJ/kg(0.75)), irrespective of cow group. We concluded, therefore, that cow groups evaluated in the present study had no significant effects on energy expenditure or energetic efficiency. However, the maintenance energy requirement (MJ/kg(0.75)) was not constant (as adopted in the majority of energy rationing systems across the world) but increased with increasing feed intake.

Effects of diet forage proportion on maintenance energy requirement and the efficiency of metabolizable energy use for lactation by lactating dairy cows.

The objective of the present study was to examine the effect of dietary forage proportion (FP) on metabolizable energy (ME) requirement for maintenance (MEm) and the efficiency of ME use for lactation (kl) in lactating dairy cows. Data used were derived from 32 calorimetric chamber experiments undertaken at our institute between 1992 and 2010, including data from 818 Holstein-Friesian cows (HF), 50 Norwegian Red cows, and 62 crossbred cows (Jersey × HF or Norwegian Red × HF). Animals were offered forage-only rations (n=66) or forage and concentrate rations (n=864) with FP ranging from 18 to 100% (dry matter basis). The effect of FP was evaluated by dividing the whole data set into 4 groups according to the FP ranges, categorized as FP <30%, FP=30 to 59%, FP=60 to 99%, and FP=100%. The MEm for individual cows was calculated from heat production minus energy losses from inefficiencies of ME use for lactation, energy retention and pregnancy, and kl was obtained from milk energy output adjusted to zero energy balance (El(0)) divided by ME available for production. Increasing FP significantly reduced ME intake and milk energy output, although the differences between the 2 low FP groups were not significant. However, increasing FP significantly increased the ratio of heat production over ME intake and MEm (MJ/kg(0.75)), with the exception that the increases did not reach significance in heat production/ME intake between FP <30% and FP=30 to 59%, or in MEm between FP=60 to 99% and FP=100%. However, the FP had no significant effect on the kl values, which were similar among the 4 groups of cows. The effect of FP was also evaluated using the linear mixed regression technique relating El(0) to ME intake. The results demonstrated that with a common regression coefficient (slope), the regression constants (intercepts) taken as net energy requirement for maintenance significantly increased with increasing FP. However, the increase between the 2 high FP groups did not research significance. It is concluded that increasing diet FP had no effects on kl but significantly increased maintenance energy requirement (MJ/kg(0.75)). These results indicate that using the current energy feeding systems to ration dairy cows managed under low input systems may underestimate their nutrient requirements, because the majority of feeding systems adopted globally do not differentiate the maintenance energy requirements between low and high forage input systems.

The effect of urea and ammonia treatments on the survival of Salmonella spp. and Yersinia enterocolitica in pig slurry.

AIMS: The objective of this study was to investigate the survival of Salmonella and Yersinia enterocolitica strains in pig slurry and evaluate urea and ammonia as disinfection strategies. METHODS AND RESULTS: Salmonella Anatum, Salmonella Derby, Salmonella Typhimurium DT19 and Y. enterocolitica bioserotypes 4, O:3, 2, O:5,27 and 1A, O:6,30 were selectively marked by insertion of the plasmid, pGLO encoding for green fluorescent protein and for ampicillin resistance. Strain cocktails were inoculated into fresh pig slurry (control), slurry treated with urea [final concentration 2% w/w, (0.33 mol l(-1) )] and slurry treated with ammonia [final concentration 0.5% w/w, (0.3 mol l(-1) )] and stored at 4, 14 and 25°C. Bacterial counts were determined at regular intervals on xylose lysine deoxycholate agar (XLD), and XLD supplemented with ampicillin (0.1 mg ml(-1) ) and arabinose (0.6 mg ml(-1) ) for Salmonella and cefsulodin-irgasan-novobiocin agar (CIN) and CIN supplemented with ampicillin and arabinose for Y. enterocolitica. The pH of the control-, urea- and ammonia-treated samples ranged from 7.1 to 7.7, 8.8 to 8.9 and 8.0 to 8.3, respectively. Salmonella D(4) values ranged from 2.71 to 21.29 days, D(14) values from 2.72 to 11.62 days and D(25) values from 1.76 to 6.85 days. The equivalent D values ranges for the Y. enterocolitica strains were 3.7-19.23, 1.8-16.67 and 1.63-7.09 days, respectively. Treatment significantly (P < 0.01) affected D values with control > ammonia > urea, as did incubation temperature; 4 > 14 > 25°C. CONCLUSIONS: Urea and to a lesser extent ammonia may be used to disinfect Salmonella- and/or Y. enterocolitica-contaminated pig slurry, decreasing the storage time required while increasing its fertilizer value. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents data supporting the treatment of pig slurry to kill important zoonotic agents, thereby reducing environmental contamination, cross-infection of other animals and decreasing zoonotic disease in the food chain.

Transfer of verocytotoxigenic Escherichia coli O157, O26, O111, O103 and O145 from fleece to carcass during sheep slaughter in an Irish export abattoir.

The purpose of this study was to investigate carriage and transfer of verocytotoxigenic Escherichia coli (VTEC) O157, O26, O111, O103 and O145 from fleece to dressed carcasses of 500 sheep, and to establish the virulence potential of recovered VTEC. Individual sheep were tracked and sampled (10 g fleece, full carcass swab) through the slaughter process. Samples were examined for the presence of verotoxin (vt1 and vt2) genes using a duplex real-time PCR assay and positive samples were further screened for the presence of the above five serogroups by real-time PCR. VTEC cells were recovered from PCR positive samples by serogroup specific immunomagnetic separation and confirmed by serogroup specific latex agglutination and PCR. Isolates were subject to a virulence screen (vt1, vt2, eaeA and hlyA) by PCR and isolates carrying vt genes were examined by Pulsed-Field Gel Electrophoresis (PFGE). VTEC O26 was recovered from 5/500 (1.0%) fleece and 2/500 (0.4%) carcass samples. VTEC O157 was isolated from 4/500 (0.8%) fleece samples and 3/500 (0.6%) carcass samples. E. coli O103 was recovered from 84/500 (16.8%) fleece and 68/500 (13.6%) carcasses, but only one E. coli O103 isolate (0.2%) carried vt genes. E. coli O145 was recovered from one fleece sample, but did not carry vt genes. E. coli O111 was not detected in any samples. For the four serogroups recovered, the direct transfer from fleece to carcass was not observed with PFGE showing that VTEC O26 isolates from a matched fleece/carcass "pair" were not identical. This study shows that while VTEC O157 are being carried by sheep presented for slaughter in Ireland, other potentially clinically significant verotoxin producing strains (particularly VTEC O26) are emerging.

Chlamydia pneumoniae antibodies and C-reactive protein levels in patients with abdominal aortic aneurysms.

INTRODUCTION: The study aim was to assess the relationship between the presence of antibodies to Chlamydia pneumoniae and abdominal aortic aneurysm (AAA) incidence. PATIENTS AND METHODS: Consecutive AAA patients and AAA-free controls were recruited prospectively. Serum samples from both groups were examined to determine Immunoglobulin (Ig) A and IgG titres against Chlamydia pneumoniae by ELISA and C-reactive protein (CRP) concentrations. Results were expressed as mean (SD) or median (IQR) and compared using χ (2) and Mann-Whitney U tests. A P value of <0.05 was considered statistically significant. RESULTS: Each study group (AAA/nAAA) comprised 250 patients. 196 (78.7%) AAA patients had positive IgA antichlamydial antibody titres, compared to 181 (72.4%) in the control group (P = 0.008, OR 2.0, 95% CI 1.2-3.5). However, positive IgG antibody titres were similar (191 versus 203; P = 0.222, OR 0.7, 95% CI 0.4-1.3). Average CRP concentrations were higher in AAA individuals. IgA or IgG antibody titres were not related to CRP concentrations. CONCLUSIONS: These results demonstrated that the frequent incidence of Chlamydia pneumoniae antibodies within the general population makes it difficult to relate its presence to AAA development, despite the high IgA antibody titres. In addition, raised CRP concentrations in AAA patients are not related to the presence of antichlamydial antibodies.

Comparison of aerial counts at different sites in beef and sheep abattoirs and the relationship between aerial and beef carcass contamination.

The study examined and compared levels of aerial contamination in commercial beef and sheep plants at four sites, i.e. lairage, hide/fleece pulling, evisceration and chilling. Aerial contamination was determined by impaction and sedimentation onto Plate Count Agar to enumerate Total Viable Counts, MacConkey Agar to enumerate coliforms and Violate Red Bile Glucose Agar to enumerate Enterobacteriaceae. AS I cannot see any difference in the text here - I am not sure what the change is?. The levels of aerial contamination were similar at equivalent sites in beef and sheep plants, irrespective of the sampling method or the type of organisms recovered. Mean log counts recovered on each medium in the chillers were generally significantly lower (P < .05) than the corresponding mean log numbers recovered at the other three sites. The relationship between impaction (air) and sedimentation (surface) counts could be described by the surface to air ratio (SAR) which in this study had an R(2) of 0.77. Further studies in an experimental plant compared counts recovered from the neck of beef carcasses with aerial counts determined by impaction and sedimentation onto agar and irradiated meat pieces. A relationship between counts on beef carcasses and in the air could not be established, irrespective of the method used to compare counts.

Diversity of culturable psychrophilic and psychrotrophic anaerobic bacteria isolated from beef abattoirs and their environments.

This study identified 431 psychrophilic or psychrotrophic isolates from commercial Irish beef abattoir environments and "blown packs" of vacuum-packed beef, using PCR and 16S rRNA sequencing, and estimated their intraspecies genetic diversity using restriction fragment length polymorphism (RFLP) analysis and spacer region PCR (SR-PCR). Twenty-five species were identified in the 431 isolates, with the most frequently recovered species being Clostridium gasigenes (n=315), Clostridium estertheticum (n=17), and a potentially novel species designated strain TC1 (n=52). These species were previously found to be associated with a particular type of spoilage known as blown-pack spoilage (BPS), which occurs in chilled-stored (i.e., -1.5°C to 4°C) vacuum-packaged meat within 2 to 4 weeks and involves the production of large volumes of gas. Overall, the study demonstrates the considerable and not previously reported diversity of the anaerobic microflora in abattoirs and the presence of a wide range of organisms capable of causing BPS at chilled temperatures.

Incidence and survival of non-O157 verocytotoxigenic Escherichia coli in soil.

AIMS: This study estimated the incidence of non-O157 verocytotoxigenic Escherichia coli (VTEC) in farm pasture soils and investigated the survival of non-O157 VTEC in clay and sandy loam soils. METHODS AND RESULTS: Twenty farms were tested over a 12-month period by sample enrichment in tryptone soya broth plus vancomycin, followed by PCR screening for the presence of vt1 and vt2 genes. Of the 600 soil samples, 162 (27%), across all farms, were found to contain vt1 and/or vt2 genes. The enrichment cultures from the 162 PCR-positive samples were plated onto Chromocult tryptone bile X-glucuronide agar (TBX), presumptive VTEC colonies recovered, confirmed as VTEC by PCR and serotyped. Samples of the two predominant soil types in Ireland (clay and sandy) were homogenized, characterized in terms of pH, boron, cobalt, copper, potassium, magnesium, manganese, phosphorus, zinc and organic matter content, inoculated with washed suspensions of eight non-O157:H7 soil isolates and six bovine faecal isolates and stored at 10°C for up to 201 days. Inoculum survival rates were determined at regular intervals by recovering and plating soil samples on TBX. All inoculated non-O157 serotypes had highest D-values in the sandy loam soil with D-values ranging from 50·26 to 75·60 days. The corresponding range in clay loam soils was 31·60-48·25 days. CONCLUSIONS: This study shows that non-O157 VTEC occur widely and frequently in pasture soils and can persist in such environments for several months, with considerable opportunity for recycling through farm environments, and cattle, with clear potential for subsequent transmission into the human food chain. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first such study of non-O157 VTEC in farm soils and found that these VTEC are frequent and persistent contaminants in farm soils. In light of recent epidemiological data, non-O157 VTEC should be seen as an emerging risk to be controlled within the food chain.

The influence of storing beef aerobically or in vacuum packs on the shelf life of mince.

AIMS: To investigate the influence of aerobic or vacuum pack storage of beef trimmings on the microbiology, colour and odour of subsequently produced mince. METHODS AND RESULTS: Trimmings stored aerobically for 7 or 10 days and in vacuum packs for 7, 10, 14 or 22 days at 0 or 5°C were minced, stored aerobically at 0 or 5°C for up to 7 days and examined daily to determine Total viable, Pseudomonas, Lactic acid bacteria, Brochothrix thermosphacta, and Enterobacteriaceae counts, colour and odour. Mincing reduced counts, particularly of Pseudomonas, B. thermosphacta and Enterobacteriaceae, probably because of the action free radicals released from muscle and bacterial cells. Storage of vacuum-packed trimmings for 22 days resulted in improved mince colour and inhibition of the growth of Pseudomonas. CONCLUSIONS: The shelf life of mince from trimmings is directly influenced by the trimmings storage conditions, and longer-term vacuum storage of trimmings produced improvements in mince quality. SIGNIFICANCE AND IMPACT OF THE STUDY: There appears to be no scientific rationale for limiting the storage of vacuum packaging beef trimmings to 15 days, prior to mince production, as stated in EU 835/2004. This study identifies advantages in storing trimmings in vacuum packs for at least 21 days prior to mincing, in terms of improved mince quality.

The effect of storage temperature and inoculum level on the time of onset of 'blown pack' spoilage.

AIMS: To examine the effect of storage temperature and inoculum level on the time of onset of 'blown pack' spoilage (BPS) caused by psychrotolerant bacteria in vacuum-packed (VP) meats. METHODS AND RESULTS: Gas-producing species and strains (n = 11), recovered in our laboratory or reported as associated with BPS, were inoculated onto beef or lamb meat pieces at final levels of <10, 10, 10(2) and 10(3) CFU cm(-2), VP and stored at -1.5, 1 or 4 degrees C. Six strains produced observable amounts of gas within 42 days and a further four strains produced gas within 100 days. BPS was observed earliest in VP meats inoculated with Clostridium estertheticum ssp. estertheticum at all inoculum levels/storage temperature combinations examined. Storage temperature and inoculum level significantly affected (P < 0.001 and P < 0.05 respectively) the onset of BPS in all cases. CONCLUSIONS: Controlling contamination levels and lowering the storage temperature delay the onset of BPS. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrates the positive effects of low contamination-low temperature as control interventions preventing/delaying BPS in VP chilled meats and identifies some of the contaminants most likely to cause BPS in chilled stored VP meat products.

Effects of reduction in beef surface water activity on the survival of Salmonella Typhimurium DT104 during heating.

AIM: To investigate the influence of reducing beef surface water activity (a(w)) on the survival of Salmonella Typhimurium DT104 during heating. METHODS AND RESULTS: Beef discs were surface inoculated with S. Typhimurium DT104 and either untreated or dried to achieve surface a(w) values of 0.95, 0.85 and 0.70. The samples were vacuum packed, heat-treated at 60 degrees C and removed at predetermined times. The inactivation curves were influenced by a(w) and treatment time. Biphasic inactivation curves were observed for S. Typhimurium DT104 heat-treated on beef samples with altered a(w) values, which were characterized by an initial decline in cell numbers at commencement of heating followed by a much slower rate of inactivation during the remaining treatment period. Point estimates of the heating time required to achieve a 1 log reduction on beef surfaces with a(w) of 0.99, 0.95, 0.85 and 0.70 were 0.5, 1.55, 11.25 and 17.79 min, respectively. CONCLUSIONS: A decrease in beef surface a(w) can substantially enhance the survival of S. Typhimurium DT104 after heating. SIGNIFICANCE AND IMPACT OF THE STUDY: Caution needs to be taken using dry air as a decontamination method as this may rapidly decrease product surface and pathogen a(w) values resulting in enhanced survival.

Isolation and sources of 'blown pack' spoilage clostridia in beef abattoirs.

AIMS: (i) To evaluate methods for isolation and molecular detection of blown pack spoilage (BPS) clostridia and (ii) to survey beef abattoirs for sources and distributions of Clostridium estertheticum and Cl. gasigenes. METHODS AND RESULTS: Molecular detection and conventional isolation methods were used to detect and recover BPS associated clostridia (Cl. estertheticum and Cl. gasigenes), from four commercial Irish beef abattoirs and their environments, during a one year study. DNA-based methods detected 218 Cl. estertheticum and 300 Cl. gasigenes, from 1680 samples, whereas culture-methods only yielded 17 Cl. estertheticum and 176 Cl. gasigenes isolates. BPS Clostridia were frequently detected in beef abattoirs and their environments, especially at areas prior to hide removal. The study noted a higher percentage of positive samples during the month of May (38.6%). CONCLUSIONS: (i) DNA-based techniques are the most reliable ways to determine the presence of these organisms in various samples and (ii) hides and faeces are the main reservoirs of BPS clostridia in the abattoirs. SIGNIFICANCE AND IMPACT OF THE STUDY: This paper provides useful information to detect BPS organisms, as well as to develop a science-based control strategy of the problem.

Changes in Escherichia coli O157:H7 numbers during holding on excised lean, fascia and fat beef surfaces at different temperatures.

AIM: To investigate changes in Escherichia coli O157:H7 numbers on excised beef carcass surfaces over 72 h at different temperatures. METHODS AND RESULTS: Excised lean meat, fascia and fat were inoculated with E. coli O157:H7 and held in an environmental chamber for 72 h, at air speed 0.5 m s(-1), relative humidity (RH) 90%, and temperatures 4, 8 and 12 degrees C. On lean, pathogen counts increased significantly at 12 degrees C. On fascia, significant reductions in counts occurred at 4 and 8 degrees C. Pathogen numbers were significantly reduced on fat at 4, 8 and 12 degrees C (64 h). Counts on fat were significantly less at all temperatures, compared to lean or fascia and surface water activity, a(w), decreased significantly over time on fat at 4 degrees C. Significant decreases in surface pH values were recorded on all meat substrates. CONCLUSIONS: The survival of E. coli O157:H7 varied in relation to the meat substrate and the holding temperature. Reductions in counts on fat surfaces appeared to be related to low surface a(w) values. No relationship between pathogen survival and surface pH was established. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of excised meat pieces in an environmental cabinet offers a more flexible approach to determining the use of different chilling regimes in the production of safe meat.

The survival of Salmonella enterica serovar Typhimurium DT104 and total viable counts on beef surfaces at different relative humidities and temperatures.

AIMS: The aim of this study was to investigate changes in Salmonella and total viable count (TVC) survival on beef carcass surfaces stored for 72 h under different combinations of relative humidity (i.e. RH 75% or 96%) and temperature (5 degrees C or 10 degrees C). METHODS AND RESULTS: The influence of low water activity (a(w)) and temperature on the survival and growth of Salmonella enterica serovar Typhimurium DT104 and the aerobic mesophilic flora on meat pieces from different sites on beef carcasses was investigated, under controlled conditions (75% or 96% RH; 5 or 10 degrees C) in an environmental cabinet. Salmonella counts declined during storage at low a(w) (75% RH) conditions at 5 degrees C or 10 degrees C. Salmonella counts increased during storage at high a(w) (96% RH) at 10 degrees C only. At 5 degrees C, TVCs increased during storage at high a(w), but not at low a(w). TVCs increased on all samples from carcasses stored at high or low a(w) at 10 degrees C, except those samples taken from areas of surface fat. CONCLUSIONS: This suggests that substrate composition dictates growth rates under low a(w) conditions. The results are discussed in terms of the possible protective effects of substrate osmolyte accumulation in bacterial survival and/or growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The data obtained in this study provides useful insights on the influence of a(w) and temperature on pathogen survival on meat surfaces at chill temperature.

Molecular characterisation of the recA locus in clinical isolates of verocytotoxigenic E. coli O157:H7.

Molecular epidemiology of verocytoxigenic Escherichia coli O157:H7 is important to help elucidate reservoirs and modes of transmission, particularly between animals and humans. As the recA gene locus is now beginning to gain application in bacterial genotyping schemes, and as it has not been examined previously in E. coli O157 isolates, this study aims to examine potential polymorphic variation as a possible epidemiological marker for the subspecies characterisation of clinically significant verocytotoxigenic E. coli O157:H7. A novel polymerase chain reaction (PCR) assay was designed to target a 638 bp region of the recA gene in E. coli O157 isolates. The PCR amplification of genomic DNA from extracted organisms was able to generate an amplicon of the expected size (approximately 638 bp) for all E. coli O157:H7 examined (n=80), as well as for other non-O157 E. coli and other members of the Enterobacteriaeceae including Citrobacter, Hafnia, Shigella, Enterobacter and Providencia. Subsequent restriction fragment length polymorphism (RFLP) and single-stranded conformational polymorphism (SSCP) analyses of these recA amplicons were able to differentiate E. coli O157 from the organisms examined, but were unable to distinguish between 79 isolates of wild-type E. coli O157, suggesting a highly conserved recA gene structure within the local population of organisms examined.

Survival of Escherichia coli O157:H7 and non-pathogenic E. coli on irradiated and non-irradiated beef surfaces.

This study examined changes in numbers of pathogenic (PEC) and non-pathogenic (NPEC) Escherichia coli during storage at 10°C on the surfaces of irradiated (IR) and non-irradiated (NIR) meat pieces excised from the neck, brisket and rump of beef carcasses and in Brain Heart Infusion Broth (BHI) and Maximum Recovery Diluent (MRD). On irradiated meat pieces, there were significant differences between mean PEC and NPEC counts at all sites. Differences in counts were also observed between IR and NIR surfaces and among the three meat sites for both E. coli types. These differences occurred only on IR samples, suggesting that the irradiation associated reductions in normal beef surface flora influenced survival of both E. coli types. PEC and NPEC counts increased during storage in BHI, but only NPEC counts increased in MRD. The results of this study highlight the impact of meat surface type and the presence/absence of the normal beef carcass surface flora on E. coli survival and/or growth during meat storage. Such previously unreported effects, and their precise mechanisms, have direct implications in the development and application of accurate models for the prediction of the safety and shelf life of stored meat.

Transfer of ampicillin resistance from Salmonella Typhimurium DT104 to Escherichia coli K12 in food.

AIMS: To investigate the transfer of antibiotic resistance from a donor Salmonella Typhimurium DT104 strain to a recipient Escherichia coli K12 strain. METHODS AND RESULTS: Mating experiments were conducted in broth, milk and ground meat (beef) at incubation temperatures of 4, 15, 25 and 37 degrees C for 18 and 36 h. Ampicillin-resistance transfer was observed at similar frequencies in all transfer media at 25 and 37 degrees C (10(-4) to 10(-5) log(10 )CFU ml g(-1), transconjugants per recipient) for 18 h. At 15 degrees C, transfer was observed in ground meat in the recipient strain (10(-6), log10 CFU g(-1), transconjugants per recipient), but not in broth or milk. At 4 degrees C, transfer did not occur in any of the examined mediums. Further analysis of the E. coli K12 nal transconjugant strain revealed the presence of a newly acquired plasmid (21 kbp) bearing the beta-lactamase gene bla(TEM). Transconjugants isolated on the basis of resistance to ampicillin did not acquire any other resistant markers. CONCLUSION: This study demonstrates the transfer of antibiotic resistance in food matrices at mid-range temperatures. SIGNIFICANCE AND IMPACT OF THE STUDY: It highlights the involvement of food matrices in the dissemination of antibiotic-resistant genes and the evolution of antibiotic-resistant bacteria.

Survival of antibiotic resistant and antibiotic sensitive strains of E. coli O157 and E. coli O26 in food matrices.

Escherichia coli O157:H7 or E. coli O26, which were AS (antibiotic sensitive), AR (laboratory created antibiotic resistant mutants), or naturally MAR (multi-antibiotic resistant), were inoculated into laboratory media, yoghurt or orange juice and their growth/survival monitored during enrichment at 37 degrees C or storage at 4 degrees C. The strains were also inoculated into minced beef and their thermal inactivation (D-values) examined at 55 degrees C, with and without a prior heat shock at 48 degrees C. The growth kinetics (lag phases, growth rates) of the VTEC (verocytotoxigenic E. coli), incubated over 24 h at 37 degrees C in laboratory media, were similar regardless of the presence or absence of antibiotic resistance. In yoghurt and orange juice, E. coli O157:H7 MAR died off significantly faster (P<0.05) than any of other VTEC strains examined. E. coli O157:H7 MAR was also found to be significantly more heat sensitive (P<0.05) than the other VTEC strains tested. The reasons for the observed differences in survival of the different VTEC strains and the link between antibiotic resistance and survival in VTEC organisms are discussed.

Antimicrobial resistance in Irish isolates of verocytotoxigenic Escherichia coli (E. coli)--VTEC.

This study compared the antimicrobial resistance profiles of Escherichia coli O157:H7 isolates (n=257) recovered from bovine hides, minced beef and human clinical samples in Ireland, to those profiles of a range of Irish non-O157 E. coli (O111 and O26) isolates (n=31) from a variety of clinical and veterinary sources. Four multi-drug resistant (MDR) E. coli O157:H7 food isolates were identified, with resistance to 10 (1 isolate), 6 (1 isolate) and 4 (2 isolates) antimicrobial agents, respectively. Two of these isolates (resistant to 7 and 4 antimicrobial classes) were characterised further by molecular methods and found to contain class 1 integrons along with a beta-lactamase-encoding tem-1 gene. Transfer of antimicrobial resistance (ampicillin, streptomycin and sulphonamides), the tem-1 gene and markers (int1, qacEDelta1, sul1) characteristic of class 1 integrons were evident in one MDR isolate (resistant to 4 antimicrobial classes) when conjugation and transformation experiments were performed. A clinical isolate and a veterinary isolate of the O111 serotype were MDR and resistant to 4 and 3 antimicrobial classes, respectively. These data suggest that the prevalence of antimicrobial resistance among the three VTEC serotypes examined in this study is low. However, these organisms may become a public health risk should they enter the food chain.

Characterisation of E. coli O157 isolates from bovine hide and beef trimming in Irish abattoirs by pulsed field gel electrophoresis.

Escherichia coli O157 isolates from bovine hide (n=117) and beef trimmings (n=32) from a single abattoir were examined by pulsed field gel electrophoresis (PFGE). Using BioNumerics software, dendrograms of isolates from each sample type (i.e. hide and beef trimming) were produced. In assessing the genetic relatedness of isolates, a similarity criterion of 80% was applied. The 117 E. coli O157 hide isolates were grouped into 14 clusters, comprising of 109 different PFGE profiles. Of the 109 different PFGE profiles, 8 were common to multiple isolates (i.e. shared 100% similarity by PFGE). The 32 E. coli O157 beef trimming isolates produced 28 different PFGE profiles and 2 clusters. Of the 28 PFGE profiles, 2 were common to multiple isolates and the remaining 26 were distinct. On a number of sampling occasions, isolates displaying identical PFGE patterns were recovered from multiple isolates collected from a single sample type (i.e. hides or trimmings), suggesting cross contamination from contaminated hides/animals to uncontaminated hides/animals and from contaminated beef trimmings to uncontaminated beef trimmings during abattoir operations.