Contamination Of Meat Products Research Articles

Detection of Yersinia spp. in meat products by enrichment culture, immunomagnetic separation and nested PCR

The prevalence of Yersinia enterocolitica in meat products was assessed by four methods: cold enrichment in trypticase soy broth (A), enrichment in modified Rappaport broth at 25 °C (B), concentration by immunomagnetic separation (C) and yadA nested PCR (D). Furthermore, the pathogenic potentials of the isolates were established by phenotypic and genotypic tests, and their genomic relationships were determined by pulsed-field gel electrophoresis (PFGE). A total of 238 samples were collected at retail level in the city of San Luis, Argentina, during the period 2007–2008. The highest Yersinia prevalence in meat products was observed by method D (92 positive samples), followed by methods A (13 positive samples) and C (5 positive samples); however, no isolation was obtained by method B. Fourteen Y. enterocolitica and 4 Yersinia intermedia strains were recovered by culture. All Y. enterocolitica 2/O:9 strains gave results related to virulence by phenotypic tests and exhibited the genotype virF+myfA+ail+ystA+. Two biotype 1A strains showed a genotype virF−myfA−ail+ystA+ystB+. The 14 Y. enterocolitica strains isolated during this work plus one reference strain were separated into 11 genomic types by PFGE. This genomic heterogeneity of the isolates shows the diversity of Y. enterocolitica strains in our region. It is the first time that IMS was used to search Y. enterocolitica strains from naturally contaminated meat products.

Differential Virulence of Clinical and Bovine-Biased Enterohemorrhagic Escherichia coli O157:H7 Genotypes in Piglet and Dutch Belted Rabbit Models

Enterohemorrhagic Escherichia coli O157:H7 (EHEC O157) is an important cause of food and waterborne illness in the developed countries. Cattle are a reservoir host of EHEC O157 and a major source of human exposure through contaminated meat products. Shiga toxins (Stxs) are an important pathogenicity trait of EHEC O157. The insertion sites of the Stx-encoding bacteriophages differentiate EHEC O157 isolates into genogroups commonly isolated from cattle but rarely from sick humans (bovine-biased genotypes [BBG]) and those commonly isolated from both cattle and human patients (clinical genotypes [CG]). Since BBG and CG share the cardinal virulence factors of EHEC O157 and are carried by cattle at similar prevalences, the infrequent occurrence of BBG among human disease isolates suggests that they may be less virulent than CG. We compared the virulence potentials of human and bovine isolates of CG and BBG in newborn conventional pig and weaned Dutch Belted rabbit models. CG-challenged piglets experienced severe disease accompanied by early and high mortality compared to BBG-challenged piglets. Similarly, CG-challenged rabbits were likely to develop lesions in kidney and intestine compared with the BBG-challenged rabbits. The CG strains used in this study carried stx2 and produced significantly higher amounts of Stx, whereas the BBG strains carried the stx2c gene variant only. These results suggest that BBG are less virulent than CG and that this difference in virulence potential is associated with the Stx2 subtype(s) carried and/or the amount of Stx produced.

Analysis of the bacterial community within carrot wash water

Vegetables are washed after harvest to remove unwanted organic and inorganic particles, but wash water contaminated with certain pathogenic microorganisms can potentially contaminate produce. In this study, the microbial diversity of wash water was analyzed in samples taken from a carrot-processing facility. A 16S rRNA gene library with 427 clones was constructed and analyzed by amplified rDNA restriction analysis. For taxonomic classification, the 16S rRNA gene nucleotide sequences of 94 amplified rDNA restriction analysis fingerprints were determined. Each fingerprint indicates a distinct group of microorganisms. The nucleotide sequences were assigned to corresponding reference species. The most prevalent genus was Tolumonas , with 26% of the clones, followed by Acinetobacter and Flacobacterium , with 11% each. The latter two genera contain species that are known to cause nosocomial infections. The fourth most common genus was Arcobacter , comprising 9% of all clones. Some species of Arcobacter are considered to be emerging food pathogens, mainly associated with the contamination of meat products. So far, they have not been considered as contaminants of fresh produce. Based on the sequence data, an Arcobacter-specific PCR assay was developed to facilitate the detection of vegetable-associated Arcobacter strains.

Salting of dry-cured meat – A potential cause of contamination with the ochratoxin A-producing species Penicillium nordicum

Penicillium nordicum is a known contaminant of protein-rich foods and is primarily found on dry-cured meat products. It is an important producer of the mycotoxin ochratoxin A, which has nephrotoxic and cancerogenic activities. Recently a high number of P. nordicum strains was isolated from different dry-cured meat products from one of the Slovenian meat-processing plants. Since we have isolated P. nordicum in high counts also from Artic habitats, such as sea water and sea ice and due to its ability to grow well at low temperatures and at increased salinity, sea salt was suspected as the possible source of P. nordicum. In the present study contamination of meat products, air in the meat-processing plant and sea salt used for salting were analysed. When 50 g of salt sample from a sealed package was dissolved in sterile water and filtered, 12 colonies of P. nordicum were obtained on solid medium incubated at 15 °C, while a salt sample from an open vessel in the meat-processing area developed high, uncountable number of colonies. Amplified fragment length polymorphism analyses of P. nordicum isolates from different sources showed that contamination of meat products via salt was possible. Three selected isolates examined for extrolites all produced ochratoxin A. As contamination of dry-cured meat products with P. nordicum represents a potential health risk for consumers and workers in the meat-processing plants, salt should be taken into account as a potential cause of such contaminations.

Chemical sensing in mammalian host-bacterial commensal associations.

The mammalian gastrointestinal (GI) tract is colonized by a complex consortium of bacterial species. Bacteria engage in chemical signaling to coordinate population-wide behavior. However, it is unclear if chemical sensing plays a role in establishing mammalian host-bacterial commensal relationships. Enterohemorrhagic Escherichia coli (EHEC) is a deadly human pathogen but is a member of the GI flora in cattle, its main reservoir. EHEC harbors SdiA, a regulator that senses acyl-homoserine lactones (AHLs) produced by other bacteria. Here, we show that SdiA is necessary for EHEC colonization of cattle and that AHLs are prominent within the bovine rumen but absent in other areas of the GI tract. We also assessed the rumen metagenome of heifers, and we show that it is dominated by Clostridia and/or Bacilli but also harbors Bacteroidetes. Of note, some members of the Bacteroidetes phyla have been previously reported to produce AHLs. SdiA-AHL chemical signaling aids EHEC in gauging these GI environments, and promotes adaptation to a commensal lifestyle. We show that chemical sensing in the mammalian GI tract determines the niche specificity for colonization by a commensal bacterium of its natural animal reservoir. Chemical sensing may be a general mechanism used by commensal bacteria to sense and adapt to their mammalian hosts. Additionally, because EHEC is largely prevalent in cattle herds, interference with SdiA-mediated cattle colonization is an exciting alternative to diminish contamination of meat products and cross-contamination of produce crops because of cattle shedding of this human pathogen.

Phage Therapy To Reduce PreprocessingSalmonellaInfections in Market-Weight Swine

Contamination of meat products with food-borne pathogens usually results from the carcass coming in contact with the feces of an infected animal during processing. In the case of Salmonella, pigs can become colonized with the organism during transport and lairage from contaminated trailers and holding pens, resulting in increased pathogen shedding just prior to processing. Increased shedding, in turn, amplifies the likelihood of carcass contamination by magnifying the amount of bacteria that enters the processing facility. We conducted a series of experiments to test whether phage therapy could limit Salmonella infections at this crucial period. In a preliminary experiment done with small pigs (3 to 4 weeks old; 30 to 40 lb), administration of an anti-Salmonella phage cocktail at the time of inoculation with Salmonella enterica serovar Typhimurium reduced Salmonella colonization by 99.0 to 99.9% (2- to 3-log reduction) in the tonsils, ileum, and cecum. To test the efficacy of phage therapy in a production-like setting, we inoculated four market-weight pigs (in three replicates) with Salmonella enterica serovar Typhimurium and allowed the challenged pigs to contaminate a holding pen for 48 h. Sixteen naïve pigs were randomly split into two groups which received either the anti-Salmonella phage cocktail or a mock treatment. Both groups of pigs were comingled with the challenged pigs in the contaminated pen. Treatment with the anti-Salmonella phage cocktail significantly reduced cecal Salmonella concentrations (95%; P<0.05) while also reducing (numerically) ileal Salmonella concentrations (90%; P=0.06). Additional in vitro studies showed that the phage cocktail was also lytic against several non-Typhimurium serovars.

Diversity of Leuconostoc gasicomitatum associated with meat spoilage

Leuconostoc gasicomitatum isolates ( n = 384) associated with spoilage of meat and vegetable-based foods were characterised by pulsed-field gel electrophoresis (PFGE) typing. Our aim was to evaluate the diversity and distribution of spoilage-associated L. gasicomitatum isolates from meat products, and to determine whether the PFGE genotypes are specific to product, producer, or isolation year (1997-2007). PFGE typing differentiated the isolates into 68 genotypes, and revealed that none one of the 54 genotypes associated with meat products was recovered from vegetable-based foods. Generally, the meat-derived genotypes were not specific to meat animal species, and many genotypes included isolates from products of different types or processors, as well as isolates collected in different years. Furthermore, certain genotypes were repeatedly identified from products of the same processing plant suggesting that the processing environment may have an impact on L. gasicomitatum contamination of meat products.

Detection of toxigenic Clostridium difficile in pig feces by PCR

: &lt;I&gt;Clostridium difficile&lt;/I&gt; is considered an important uncontrolled cause of neonatal diarrhea. Also, the presence of bacteria in the feces of the animal could represent a zoonosic risk for the contamination of meat products. Therefore, it is necessary to have procedures available for the early detection of &lt;I&gt;C. difficile&lt;/I&gt; in animals. The current study describes a new semi-automated procedure for the recovery of &lt;I&gt;C. difficile&lt;/I&gt; DNA from pig feces and subsequent amplification by polymerase chain reaction (PCR) of three different sequences: the triose phosphate isomerase gene &lt;I&gt;tpi&lt;/I&gt;, specific for this bacterial species, and the &lt;I&gt;tcdA&lt;/I&gt; and &lt;I&gt;tcdB&lt;/I&gt; genes, which code for the A and B toxins of &lt;I&gt;C. difficile&lt;/I&gt;, respectively. Twenty-two fecal samples microbiologically positive for &lt;I&gt;C. difficile&lt;/I&gt; were used. The &lt;I&gt;tpi&lt;/I&gt; and &lt;I&gt;tcdA&lt;/I&gt; genes were amplified in all of them. The internal fragment of &lt;I&gt;tcdB&lt;/I&gt; was detected from 21 of these extracts; the negative sample gave a positive result when a different primer pair was used. None of the 10 DNA extracts obtained from culture-negative samples gave a positive result. The method presented in this article eliminates the interference caused by the possible presence of PCR inhibitors. To the authors’ knowledge, this is the first description of a PCR procedure for detection of &lt;I&gt;C. difficile&lt;/I&gt; DNA from domestic animal feces.

Enumeration and identification of Campylobacter species in the liver and bile of slaughtered cattle

Healthy cattle are considered as reservoirs for a variety of Campylobacter species. To control the bacterial contamination in meat products, quantitative assessment of campylobacters in liver and gallbladder was carried out at an abattoir. Liver and bile samples were collected from 108 healthy beef cattle after evisceration and viable counts of campylobacters were determined by a direct-plating technique using modified Cefoperazone Charcoal Deoxycholate agar (mCCDA). The suspected colonies on the highest dilution plates were subjected to biochemical tests and PCR for identification. Campylobacter species were isolated from 49 (45%) bile and 6 (5%) liver specimens examined. Numbers of campylobacters in bile and liver ranged from log 103 to 7 (median 5) and log 10 1 to 2 (median 1) cfu per ml and per g, respectively. These Campylobacter species were identified as C. fetus, C. jejuni, and C. coli. Multiple infections involving two species were observed in 16 cattle. C. fetus and C. jejuni were the predominant species in bile. Growth of C. fetus, C. jejuni, and C. coli in spiked bile samples revealed an initial exponential growth phase followed by a period with no apparent increase in colony count for 28 days. It appeared that these campylobacters can survive in bile for a long period. To determine transfer route of bacterial cells to the gallbladder, C. jejuni, C. fetus, or C. coli was inoculated intravenously in mice. The inoculated cells were recovered from bile, suggesting that the organism was transferred from the blood stream to bile duct in the liver. From these results, bile in cattle is considered to be an important contamination source of Campylobacter species in processing plants.

Isolation and characterization of methicillin-resistantStaphylococcus aureus from livestock and poultry meat

Meat samples from sheep, bovine, camel and poultry were collected from Amman area and were processed and tested for the presence of methicillin (oxacillin) resistantStaphylococcus aureus (MRSA). Identity ofS. aureus was ensured by Gram-staining and a battery of biochemical tests. From 1260 meat samples, 157S. aureus positive isolates were identified. Of the 157 isolates, 30 were resistant to methicillin levels greater than 2 μg/ml and only 15 weremecA-positive MRSA originating mainly from sheep and chicken. Subjecting themecA-positive MRSA to antibiotic susceptibility testing revealed that all isolates were resistant to β-lactam antibiotics (ampicillin, penicillin, and oxacillin) and were sensitive to vancomycin, trimethoprim, chloramphenicol and cephalothin. Randomamplified polymorphic DNA (RAPD) analysis ofmecA-positive animal isolates generated six different patterns. Comparing these results with results of isolates of human origin of our laboratory there is some molecular epidemiological relatedness between both and could be a possible source of infections through consuming contaminated meat products, direct contact or meat processing.

Post-processing application of chemical solutions for control of Listeria monocytogenes, cultured under different conditions, on commercial smoked sausage formulated with and without potassium lactate–sodium diacetate

This study evaluated post-processing chemical solutions for their antilisterial effects on commercial smoked sausage formulated with or without 1.5% potassium lactate plus 0.05% sodium diacetate, and contaminated (approximately 3–4 log cfu/cm 2) with 10-strain composite Listeria monocytogenes inocula prepared under various conditions. Inoculated samples were left untreated, or were immersed (2 min, 25±2 °C) in solutions of acetic acid (2.5%), lactic acid (2.5%), potassium benzoate (5%) or Nisaplin ® (0.5%, equivalent to 5000 IU/ml of nisin) alone, and in sequence (Nisaplin followed by acetic acid, lactic acid or potassium benzoate), before vacuum packaging and storage at 10 °C (48 days). Acetic acid, lactic acid or potassium benzoate applied alone reduced initial L. monocytogenes populations by 0.4–1.5 log cfu/cm 2, while treatments including Nisaplin caused reductions of 2.1–3.3 log cfu/cm 2. L. monocytogenes on untreated sausage formulated with antimicrobials had a lag phase duration of 10.2 days and maximum specific growth rate ( μ max) of 0.089 per day, compared to no lag phase and μ max of 0.300 per day for L. monocytogenes on untreated product that did not contain antimicrobials in the formulation. The immersion treatments inhibited growth of the pathogen for 4.9–14.8 days on sausage formulated without potassium lactate–sodium diacetate; however, in all cases significant ( P < 0.05 ) growth occurred by the end of storage. The antilisterial activity of chemical solutions was greatly enhanced when applied to product formulated with antimicrobials; growth was completely inhibited on sausage treated with acetic or lactic acid alone, and in sequence with Nisaplin. In general, habituation (15 °C, 7 days) of L. monocytogenes cells, planktonically or as attached cells to stainless-steel coupons in sausage homogenate prior to contamination of product, resulted in shorter lag phase durations compared with cells cultivated planktonically in a broth medium. Furthermore, when present, high levels of spoilage flora were found to suppress growth of the pathogen. Findings of this study could be useful to US meat processors in their efforts to select required regulatory alternatives for control of post-processing contamination in meat products.

Inactivation of transmissible spongiform encephalopathy agents in food products by ultra high pressure–temperature treatment

Bovine spongiform encephalopathy (BSE) contamination of the human food chain most likely resulted from nervous system tissue in mechanically recovered meat used in the manufacture of processed meats. The availability of effective decontamination methods for products considered at risk for BSE or other transmissible spongiform encephalopathies (TSEs) would be an attractive safeguard to human health, but neither of the two proven inactivating methods, autoclaving or exposure to strong alkali or bleach, are applicable to foodstuffs. Ultra high pressure–temperature treatment of foods is an effective decontamination method that can reduce the pathogen load while keeping unaltered the nutritional and organoleptic properties of the product. The application of different combinations of high pressure–temperature pulses to meat products ‘spiked’ with the agents of TSEs can reduce the level of infectivity by 10 3 to 10 6 mean lethal doses (LD 50) per gram of tissue. These data indicate that the high pressure–temperature treatment is a ready-to-use and feasible strategy to reduce the risk of TSEs transmission via contaminated meat products.

Next Day Detection of Salmonella spp. In Naturally Contaminated Meat Products Using Molecular Approach

Salmonella species are a well known food borne pathogen in developed and developing countries with a wide variety of illnesses conditions. The aim of this study was to compare a traditional isolation and identification method for Salmonella spp from meat products with a newer DNA/rRNA gene hybridization method [LUMIprobe 24 Salmonella] under the routine food laboratory conditions. A total of 133 samples [30 pork sausage, 40 chicken meat coated with oil and spices, 40 kebab and 23 beef burger] were collected from February to May 2004, samples were processed according to the guideline of the British Standard Cultural Method; BS EN 12824:1998, as well as, the DNA/rRNA method as guided by the manufacture. Results showed that the two methods gave equivalent sensitivity and selectivity for Salmonella spp detection. Significant time saving was achieved using the LUMIprobe 24 test allowing earlier release of results on the next day and the new molecular method experienced a reasonable level of flexibility and labor saving.

Does Food Safety Information Impact U.S. Meat Demand?

A theoretical model of consumer response to publicized food safety information on meat demand is developed with an empirical application to U.S. meat consumption. Evidence is found for the existence of pre-committed levels of consumption, seasonal factors, time trends, and contemporaneous own- and cross-commodity food safety concerns. The average demand response to food safety concerns is small, especially in comparison to price effects, and to previous estimates of health related issues. This small average effect masks periods of significantly larger responses corresponding with prominent food safety events, but these larger impacts are short-lived with no apparent food safety lagged effects on Food safety concerns in the United States have dramatically increased in the past decade with regard to incidences of contaminated meat products. Concerns have arisen because contaminated meat products can result in serious risk to the well being and health of consumers. Contamination comes from a myriad of sources, including but not limited to, outbreaks of Listeria monocytogenes, Escherichia coli (E. coli), and Salmonella (Centers for Disease Control and Prevention). Food safety problems are not isolated to the United States as other unsafe contaminates in meats have emerged across the world, including highly publicized outbreaks of Bovine Spongiform Encephalopathy (BSE) in Europe. The potential impacts of publicized food safety events on consumer’s demand for meat products in the United States include own-effects on the demand for the contaminated meat involved as well as cross-effects impacting the demand for other meats. The objective of this article is to investigate whether publicized food safety concerns surrounding beef, pork, and poultry (chicken and turkey) have impacted meat consumption. Food safety indices are constructed separately for beef, pork, and poultry

Growth dynamics of Salmonella enterica strains on alfalfa sprouts and in waste seed irrigation water.

Alfalfa sprouts and other seed sprouts have been implicated in numerous outbreaks of salmonellosis. The source of these epidemics appears to have been low-level contamination of seeds by Salmonella bacteria that developed into clinically significant populations during the seed germination process. To test the possibility that Salmonella enterica strains carry host range determinants that allow them to grow on alfalfa, strains isolated from alfalfa or other sources were surveyed for their ability to grow on germinating alfalfa seeds. An S. enterica serovar Cubana strain originally isolated from contaminated alfalfa sprouts multiplied most rapidly during the initial 24 h of the seed germination process. Germinating alfalfa seeds supported the multiplication of S. enterica cells prior to the emergence of the root radicle at 72 h. Thereafter, much lower rates of multiplication were apparent. The ability of S. enterica to grow on germinating alfalfa seeds was independent of the serovar, isolation source, or virulence of the strain. Isolates obtained from alfalfa attained population levels similar to those observed for strains isolated from contaminated meat products or stools. Each of the strains could be detected in the waste irrigation water, with populations being strongly correlated with those detected on the germinating alfalfa seeds. The S. enterica strains were capable of utilizing the waste irrigation water as a sole carbon and nitrogen source. S. enterica strains thus appear to grow saprophytically on soluble organics released from seeds during early phases of germination. The ability to detect S. enterica in the waste irrigation water early in the germination process indicates that this method may be used as a simple way to monitor the contamination of sprouts during commercial operations.

Role of variant Creutzfeldt-Jakob disease for safety of treatment with blood components: screening of lymphatic tissue is a potential tool for risk assessment.

Transmissible spongiform encephalopathies (TSEs) are neuropathological diseases caused by prions. Prions are infectious particles (PrPSc) which can induce bovine spongiform encephalopathy and most likely also the related infectious disease, variant of Creutzfeldt-Jakob disease (vCJD). The exposure of humans to orally ingested BSE agent in contaminated meat products presumably led to the emergence of vCJD. In vCJD, prions can be detected immunohistochemically not only in neuronal tissue but also in lymphoreticular tissue. vCJD is of significance in transfusion medicine because of the hypothetical transmission of prions by blood products. An immunohistochemistry method was used to allow screening for vCJD in human lymphoreticular tissue. PrPSc can be detected in the cerebrum and cerebellum of patients with sporadic Creutzfeldt-Jakob disease (sCJD) and in the lymph nodes, tonsils, and spleen of vCJD patients. This method has the major advantage of working in fixed specimens which are routinely saved in departments of pathology and therefore allows screening of large numbers of archived human lymphoreticular tissues in different regional areas and from different time points. Scrapie-positive lymphoreticular sheep tissue reacts similarly to human tissue of vCJD affected patients and is available in sufficient amounts to be used as positive control in screening programs. A method is provided which is a feasible tool for an epidemiological screening program to assess the prevalence of the assumed infectious agent of vCJD, PrPSc, in various populations.

The effects of level of bedding provision and time in lairage on the contamination of hides of finished cattle with potentially zoonotic bacteria

Many cases of food-borne illness in the UK are related to the consumption of contaminated meat products. This has highlighted the importance of adopting hygienic procedures throughout the meat production chain, including the farm and abattoir environment (Pennington, 2000). Many factors are known to affect the hygienic condition of finished cattle (Davies et al., 2000) and various husbandry practices may be used to improve cleanliness at slaughter. However, the extent to which abattoir practices contribute to the contamination of cattle hides is not known. Overnight lairage of cattle prior to slaughter is a common practice in the UK, but the extent to which this affects bacterial contamination of the hide remains to be determined. This study investigated the effects of providing additional straw bedding during lairage on the microbiological cleanliness of cattle during overnight lairage.

BSE uncertainties drag on

A recent mix-up in studies on brain tissue of animals has set back Britain's efforts to determine human exposure to the BSE agent through contaminated meat products. Nigel Williams reports.

Comparison of culture, PCR and immunoassays for detecting Escherichia coli O157 following enrichment culture and immunomagnetic separation performed on naturally contaminated raw meat products

The aims of this study were (i) to evaluate the specificity and sensitivity of three previously described PCR assays for the detection of Escherichia coli O157 and, (ii) to compare PCR, culture, and two visual immunoassays (VIAs), BioSign™ and Path-Stik™, for detecting E. coli O157 after enrichment culture and immunomagnetic separation (IMS) performed on various naturally contaminated raw beef, lamb and mixed meat products. Twelve sorbitol nonfermenting (SNF) verocytotoxin-producing (VT+) E. coli O157, 6 SNF VT− E. coli O157, 4 sorbitol fermenting (SF) VT+ E. coli O157, 3 SF VT− E. coli O157, 23 E. coli belonging to 17 other serogroups and 12 organisms of other species were used to check the specificity of PCR reactions. Only one primer pair generated amplimers only with E. coli O157 and was used for all subsequent work. After enrichment culture and on inoculated minced beef samples, PCR was as sensitive as culture for detecting 9 of the 12 strains of E. coli O157, but up to 4 log 10 more sensitive than culture for detecting three strains. Of the 120 samples of naturally contaminated meat products examined, 80 (67%) were positive by PCR, 70 (58%) were positive by BioSign™, 69 (58%) were positive by culture and 67 (56%) were positive by Path-Stik™. Although both VIAs lacked sensitivity when compared to PCR, both compared favourably with culture and both were extremely rapid and easy to perform, giving a result in less than 15 min. Eleven samples were positive by PCR and both VIAs, but negative by culture because culture plates were heavily overgrown with SF organisms, making detection of any E. coli O157 present impossible.

General Information and Statistics

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