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Biomedical subjects

J J Sheridan

Publications and source records attributed to J J Sheridan.

At least 19 recordsLinked to original sources

The ineffectiveness of organic acids, freezing and pulsed electric fields to control Escherichia coli O157:H7 in beef burgers.

AIMS: The objective of this study was to investigate the potential value of individual and combined applications of some GRAS (generally regarded as safe) additives with freezing and pulsed electric field (PEF) application, in reducing the risks associated with the presence of E. coli O157:H7 in beef burgers. METHODS AND RESULTS: Beef burgers, trimmings and filter paper were inoculated with E. coli O157:H7 and subjected to a range of chemical and physical treatments. Sequential application of 2% (v/v) lactic acid and freezing (at - 20 degrees C for 2 h) resulted in a decrease of approximately 6 log10 cfu cm(-1) in E. coli O157:H7, but only on filter paper. All other treatments were ineffective. CONCLUSIONS: Currently available methods for controlling E.coli O157:H7 in beef burgers during production are ineffective. SIGNIFICANCE AND IMPACT OF THE STUDY: Further research is needed to develop controls for E. coli O157:H7 during beef burger production.

Animals↗

Use of a marker organism to model the spread of central nervous system tissue in cattle and the abattoir environment during commercial stunning and carcass dressing.

Due to concerns about a link between variant Creutzfeldt-Jakob disease in humans and similar prion protein-induced disease in cattle, i.e., bovine spongiform encephalopathy (BSE), strict controls are in place to exclude BSE-positive animals and/or specified risk materials including bovine central nervous system (CNS) tissue from the human food chain. However, current slaughter practice, using captive bolt guns, may induce disruption of brain tissues and mobilize CNS tissues into the bovine circulatory system, leading to the dispersion of CNS tissues (including prion proteins) throughout the derived carcass. This project used a marker (antibiotic-resistant) strain of Pseudomonas fluorescens to model the effects of commercial captive bolt stunning procedures on the movement of mobilized CNS material within slaughtered animals and the abattoir environment. The marker organism, introduced by injection through the bolt entry aperture or directly using a cartridge-fired captive bolt, was detected in the slaughter environment immediately after stunning and in the abattoir environment at each subsequent stage of the slaughter-dressing process. The marker organism was also detected on the hands of operatives; on slaughter equipment; and in samples of blood, organs, and musculature of inoculated animals. There were no significant differences between the results obtained by the two inoculation methods (P < 0.05). This study demonstrates that material present in, or introduced into, the CNS of cattle during commercial captive bolt stunning may become widely dispersed across the many animate and inanimate elements of the slaughter-dressing environment and within derived carcasses including meat entering the human food chain.

Abattoirs↗

Beef HACCP: intervention and non-intervention systems.

While there are several generic beef HACCP documents available to the beef industry, these lack sufficient detail to be of any use other than as a general guide to HACCP. A document which clearly identifies and provides a sound scientific basis for potential critical control points (CCPs') and details critical limits, monitoring and corrective actions is clearly required. The objective of this paper is to provide such information. A detailed description of CCPs' for two different HACCP systems (an intervention and a non-intervention system) are presented and the advantages and disadvantages of each are discussed. Individual beef plants may then make an informed choice as to which HACCP system is most suitable for them and have all the specific information required for effective implementation.

Abattoirs↗

Antibiotic resistance among Listeria, including Listeria monocytogenes, in retail foods.

AIMS: In the past eight to 10 years, reports of antibiotic resistance in food-borne isolates in many countries have increased, and this work examined the susceptibility of 1001 food isolates of Listeria species. METHODS AND RESULTS: Susceptibility/resistance to eight antibiotics was determined using the Bauer-Kirby disc diffusion assay, and 10.9% of the isolates examined displayed resistance to one or more antibiotics. Resistance to one or more antibiotics was exhibited in 0.6% of Listeria monocytogenes isolates compared with 19.5% of Listeria innocua isolates. Resistance was not observed in Listeria seeligeri or Listeria welshimeri. Resistance to tetracycline (6.7%) and penicillin (3.7%) was the most frequently observed, and while resistance to one antibiotic was most common (9.1%), isolates resistant to two or more antibiotics (1.8%) were also observed. CONCLUSION: While resistance to the antibiotics most commonly used to treat human listeriosis was not observed in L. monocytogenes, the presence of such resistance in other Listeria species raises the possibility of future acquisition of resistance by L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The higher level of resistance in L. innocua compared with L. monocytogenes suggests that a species-related ability to acquire resistance to antibiotics exists.

Ampicillin↗

Thermal resistance of wild-type and antibiotic-resistant Listeria monocytogenes in meat and potato substrates.

AIMS: This study aimed to elucidate the relationship, if any, between the acquisition/possession of antibiotic resistance in strains of Listeria monocytogenes and the resistance of such strains to heat stress. METHODS AND RESULTS: D-values calculated using a linear survival model were used to compare the heat resistance of two wild-type (WT) and two antibiotic (streptomycin)-resistant (AR) mutant strains of L. monocytogenes measured in minced beef and potato substrates at 55 degrees C, with and without prior heat shock at 48 degrees C. In both minced beef and potato, no significant differences (P < 0.05) between D-values of AR and WT strains were noted. Heat shock did not significantly increase D-values of WT or AR strains in minced beef, while in potato slices, D-values in almost all cases were significantly higher in samples which had received heat-shock treatment. In minced beef, the use of a non-selective/overlay recovery medium did not result in higher D-values for any strains, while in potato, significantly higher (P < 0.05) D-values were obtained in most cases. CONCLUSION: The presence or absence of antibiotic resistance genes did not modulate the heat resistance of the strains examined in this study. SIGNIFICANCE AND IMPACT OF THE STUDY: The study demonstrated that heat shock, and the type of media used to determine bacterial numbers during heat processing, can significantly affect the D-values obtained.

Drug Resistance, Microbial↗

Comparison of selective and non-selective enrichment media in the detection of Listeria monocytogenes from meat containing Listeria innocua.

AIMS: This study investigated whether the higher incidence of recovery from meat of Listeria innocua compared with L. monocytogenes could be due to the laboratory media used, leading to an artificially lower detection of the pathogenic species, L. monocytogenes. METHODS AND RESULTS: Minced beef was inoculated with L. monocytogenes, L. innocua, or a mixture of these species, and stored at 0 or 10 degrees C under vacuum or aerobic conditions for up to 28 days. Listeria were recovered from the minced beef using selective (University of Vermont Medium, UVM) and non-selective (Buffered Peptone Water, BPW) enrichment broths after 0, 14, and 28 days of storage. In general, there were no significant differences (P < 0.05) between the numbers of L. monocytogenes recovered from minced beef samples after 24 h enrichment in BPW and the numbers recovered using UVM. In addition, the presence of L. innocua in meat samples containing L. monocytogenes did not significantly (P < 0.05) affect the numbers of L. monocytogenes recovered using either enrichment broth. In most cases there were no significant differences (P < 0.05) between the numbers of L. innocua recovered from minced beef samples after 24 h enrichment in BPW compared with numbers recovered using UVM. CONCLUSION: Listeria innocua was found to have no significant competitive advantage over L. monocytogenes in selective or non-selective enrichment media. SIGNIFICANCE AND IMPACT OF THE STUDY: The results suggest that, in some instances, the use of a selective enrichment broth offers no advantage over a non-selective enrichment broth for the recovery of Listeria species from minced beef.

Bacterial Typing Techniques↗

The survival of Escherichia coli O157:H7 in slurry from cattle fed different diets.

AIMS: The survival characteristics of Escherichia Coli O157:H7 were investigated in bovine slurry from cattle fed two different diets: (i) silage and (ii) silage + concentrates. METHODS AND RESULTS: Slurry samples collected from freshly-agitated tanks were inoculated at a level of log10 6.0 cfu g(-1) and stored in the laboratory at 10 degrees C. Over a 12 week storage period, a 3.5 and 5.5 log reduction was observed in slurry from cattle fed a silage and silage plus concentrate diet, respectively. CONCLUSIONS: The persistence of E. coli O157:H7 in slurry over a 3 month storage period indicates its potential for transmitting the organism back into the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: The discussion concludes however, that despite pathogen survival in slurry, it may not represent a major source of transmission in the farm environment.

Animals↗

Use of steam condensing at subatmospheric pressures to reduce Escherichia coli O157:H7 numbers on bovine hide.

This study used a laboratory-scale apparatus to apply subatmospheric steam to bovine hide pieces inoculated with Escherichia coli O157:H7 in maximum recovery diluent (MRD) and in high-liquid content and low-liquid content fecal suspensions (HLC fecal and LLC fecal, respectively). The survival of the organism in fecal clods, which were stored for 24 days in a desiccated state, was assessed. Inoculated fecal clods were also treated with subatmospheric steam. Steam treatment at 80 +/- 2 degrees C for 20 s reduced E. coli O157:H7 concentrations on hide inoculated to initial concentrations of approximately 7 log10 CFU/g by 5.46 (MRD inoculum), 4.17 (HLC fecal inoculum), and 5.99 (LLC fecal inoculum) log10 CFU/g. The reductions achieved in samples inoculated with LLC feces were larger than in samples inoculated with HLC feces (P < 0.05). Treatment at 80 +/- 2 degrees C for 10 s resulted in significantly smaller reductions (P < 0.05) on hide pieces of 2.54 (MRD), 1.94 (HLC fecal), and 2.15 (LLC fecal) log10 CFU/g. There were no significant differences among the reductions observed in all inoculum types in samples treated for 10 s. E. coli O157:H7 inoculated in fecal clods to 7.78 log10 CFU/g and stored at 4 or 15 degrees C survived for at least 24 days. Steam treatment (20 s) of 3-day-old clods reduced surviving E. coli O157:H7 numbers from 4.20 log10 CFU/g to below the limit of detection of the assay used (1.20 log10 CFU/g). This study shows that steam condensing at or below 80 +/- 2 degrees C can reduce E. coli O157:H7 when present on bovine hide, reducing the risk of cross contamination to the carcass during slaughter and dressing.

Animals↗

The role of microbiological testing in systems for assuring the safety of beef.

The use of microbiological testing in systems for assuring the safety of beef was considered at a meeting arranged by the International Livestock Educational Foundation as part of the International Livestock Congress, TX, USA, during February, 2000. The 11 invited participants from industry and government research organizations concurred in concluding that microbiological testing is necessary for the implementation and maintenance of effective Hazard Analysis Critical Control Point (HACCP) systems, which are the only means of assuring the microbiological safety of beef; that microbiological testing for HACCP purposes must involve the enumeration of indicator organisms rather than the detection of pathogens; that the efficacy of process control should be assessed against performance criteria and food safety objectives that refer to the numbers of indicator organisms in product; that sampling procedures should allow indicator organisms to be enumerated at very low numbers; and that food safety objectives and microbiological criteria are better related to variables, rather than attributes sampling plans.

Animals↗

Thermal inactivation of Listeria monocytogenes and Yersinia enterocolitica in minced beef under laboratory conditions and in sous-vide prepared minced and solid beef cooked in a commercial retort.

D-values were obtained for Listeria monocytogenes and Yersinia enterocolitica at 50, 55 and 60 degrees C in vacuum-packed minced beef samples heated in a laboratory water-bath. The experiment was repeated using vacutainers, which allowed heating of the beef to the desired temperature before inoculation. D-values of between 0.15 and 36.1 min were obtained for L. monocytogenes. Pre-heating the beef samples significantly affected (P < 0.05) the D60 value only. D-values for Y. enterocolitica ranged from 0.55 to 21.2 min and all the D-values were significantly different (P < 0.05) after pre-heating. In general, the D-values obtained for core inoculated solid beef samples were significantly higher (P < 0.05) than those generated in minced beef when heated in a Barriquand Steriflow commercial retort.

Animals↗

The effect of temperature and selective agents on the growth of Yersinia enterocolitica serotype O:3 in pure culture.

This study examined the individual and combined effects of the selective agents normally present in Yersinia-selective agar (i.e. cefsulodin, irgasan and novobiocin) on the growth kinetics of plasmid-bearing (P+) and plasmid-cured (P-) Yersinia enterocolitica serotype O:3 at 25 and 37 degrees C. Growth studies were carried out in pure culture, and the data obtained were subjected to linear regression analysis to determine lag phase duration(s) and growth rates of the examined strains. In general, the presence of selective agents increased the duration of the lag phase at 37 degrees C, with longer lag phases noted in all cases in which two or more selective agents were present. Growth rates in CIN broth base (CIN NA) and CIN NA plus commercial supplement (SR 109) (CIN) were faster at 37 than 25 degrees C, but in some cultures of incomplete CIN NA broth with less than three supplements added, growth tended to be faster at 25 than 37 degrees C. Generally, plasmid-bearing strains grew slower than plasmid-cured strains in most media at 37 degrees C due to virulence plasmid expression retarding growth. In some instances at 37 degrees C, it was observed that the growth rates of both plasmid-bearing and plasmid-cured strains were comparable, indicating the influence of added selective agent/s negating any effects associated with virulence plasmid expression. The effects of selective agents, incubation temperature and virulence plasmid carriage on the growth kinetics of Y. enterocolitica are discussed.

Culture Media↗

The effect of culture growth phase on induction of the heat shock response in Yersinia enterocolitica and Listeria monocytogenes.

The effect of culture growth phase on induction of the heat shock response in Yersinia enterocolitica and Listeria monocytogenes, was examined. Exponential or stationary preconditioned cultures were heat shocked and survivor numbers estimated using selective and overlay/resuscitation recovery techniques. The results indicate that prior heat shock induced increased heat resistance in both micro-organisms to higher heat treatments. Heat-shocked cells of each micro-organism were able to survive much longer than non-heat-shocked cells when heated at 55 degrees C. The size of the change in heat resistance between heat-shocked and non-heat-shocked cells was greatest for exponential cultures (X:X). Results indicate that the overall relative thermal resistance of each pathogen was dependent on cell growth phase. Stationary cultures (S:S) were significantly (P < 0.01) more thermotolerant than exponential cultures (X:X) under identical processing conditions. Under most conditions, the use of an overlay/resuscitation recovery medium resulted in higher D-values (P < 0.05) compared with a selective recovery medium.

Colony Count, Microbial↗

A membrane-immunofluorescent-viability staining technique for the detection of Salmonella spp. from fresh and processed meat samples.

A direct staining technique was investigated for the detection of viable Salmonella in fresh and processed meats. The technique involved overnight enrichment in BPW, extraction of Salmonella cells onto a polycarbonate membrane, followed by detection of the pathogen using anti-Salmonella monoclonal antibody coupled with an antibody linked-fluorescent stain (Texas Red) and a viability stain (Sytox Green). The technique was applied to the detection of Salm. enteritidis inoculated into broth culture or minced beef and then subjected to a variety of stresses including freezing (- 20 degrees C), heating (2 or 4 min at 56.9 degrees C), low pH (5 or 3.5) or high salt (2 or 4%). The correlation between traditional plate counts and the rapid count varied widely (r2 = 0.98-0.03), depending on the type and level of stress applied to the cells. The reason for the disparity in results obtained, and the potential application of the method as a diagnostic tool, are discussed.

Animals↗

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

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.

Abattoirs↗

The relationship between hide cleanliness and bacterial numbers on beef carcasses at a commercial abattoir.

Cattle were visually inspected in the lairage of a commercial abattoir and assigned to a category ranging from 1 (very clean) to 5 (very dirty) depending on the observed cleanliness of the hide. Animals from categories 2, 3 and 5 were slaughtered and total viable counts (TVCs) enumerated at five sites (hock, brisket, cranial back, bung and inside round) on the subsequent carcasses. The TVCs at the hock were significantly higher on category 5 than on category 2 carcasses (P < 0.05). Similarly, TVCs at the brisket were significantly higher on categories 3 and 5 than on category 2 carcasses (P < 0.05). There were no differences in counts among the categories at any of the other sites. The TVCs averaged over the five carcass sites were higher for category 5 than for category 2 carcasses (P < 0.05). The TVCs at the brisket were significantly higher than all other sites (P < 0.01). In general, carcasses from category 2 animals slaughtered in a batch with dirtier animals (categories 3 and 5) did not have higher TVCs than carcasses of category 2 animals slaughtered at the beginning of the day in the absence of dirtier animals. The introduction of improved hygienic practices during the dehiding of category 4 animals resulted in reduced TVCs at the brisket (P < 0.001).

Abattoirs↗

Effect of pH on survival, thermotolerance, and verotoxin production of Escherichia coli O157:H7 during simulated fermentation and storage.

Heat treatment is increasingly being introduced to fermented meat processing, since the acid tolerance properties of Escherichia coli O157:H7 can permit this organism to survive traditional processing procedures. This study investigated the effect of growth pH and fermentation on the thermotolerance at 55 degrees C of E. coli O157:H7 in a model fermented meat system. E. coli O157:H7 (strain 380-94) was grown at pH 5.6 or 7.4 (18 h at 37 degrees C), fermented to pH 4.8 or 4.4 in brain heart infusion broth, and stored for 96 h. Cells grown at pH 5.6 had higher D values at 55 degrees C (D55 degrees C) than cells grown at pH 7.4 (P < 0.001). Cells fermented to pH 4.8 had higher D55 degrees C than those fermented to pH 4.4 (P < 0.001). Cells fermented to pH 4.8 demonstrated an increase in D55 degrees C during storage (P < 0.001), whereas cells fermented to pH 4.4 showed a decrease in D55 degrees C during the same period (P < 0.001). The effect of growth pH on verotoxin production by E. coli O157:H7 was assessed using the verotoxin assay. Cells grown at pH 5.6 had lower verotoxin production then cells grown at pH 7.4. This effect was not sustained over storage. These results indicate that a lower growth pH can confer cross-protection against heat. This has implications for the production of acidic foods, such as fermented meat, during which a heating step may be used to improve product safety.

Animals↗