Raw Pork Samples Research Articles

Risk Factors and Level of Listeria monocytogenes Contamination of Raw Pork in Retail Markets in China.

Listeria monocytogenes can contaminate various foods via food processing environments and contamination of raw materials. There is a limited understanding of L. monocytogenes transmission in retail market and the role of insects in L. monocytogenes transmission in the retail environments. To better understand the risk factors of raw pork contamination, the prevalence of L. monocytogenes was examined in raw pork, retail environments and insects in a retail market over a 6-month period from March to August in 2016 in Beijing, China. A total of 2,789 samples were collected, including 356 raw pork samples, 1,392 meat contact surface swabs (MCS), 712 non-meat contact surface swabs (NMCS) and 329 insect samples. Overall, 424 (15.20%) of the samples were found to be contaminated by L. monocytogenes. Analyzed by serotyping, multilocus sequence typing and pulsed-field gel electrophoresis, the 424 L. monocytogenes isolates were divided into three serotypes (1/2c, 1/2a and 3a), 15 pulsotypes (PTs) and nine sequence types (STs), 1/2c/PT4/ST9 (244/424, 58%) was the most prevalent type of L. monocytogenes strains. The raw pork, MCS of the environments and insects were contaminated with higher levels of L. monocytogenes than NMCS of the environments, which suggested that cross contamination of L. monocytogenes between raw pork and the environment existed in the retail market, and long-term contaminated surfaces and vector insects would act as high risk factors to transmit L. monocytogenes to raw pork. Thus more effective strategies are needed to reduce the risk of retail pork meat contamination by L. monocytogenes and prevent foodborne human listeriosis.

Veterinary antibiotics in food, drinking water, and the urine of preschool children in Hong Kong

Due to the harmful effects of veterinary antibiotics (VAs) residues in food on children's health, urine samples from 31 preschool and primary school children were analyzed for 13 common VAs. Samples of raw and cooked pork, chicken, fish, milk and drinking water from the children's living areas were also analyzed for residual VAs. Urinalysis revealed one to four target antibiotics in 77.4% of the sample group, with concentrations as high as 0.36ng/mL. Norfloxacin and penicillin had the highest detection rates (48.4% and 35.5%, respectively), with median concentrations of 0.037 and 0.13ng/mL, respectively. The VA burden of children in HK was lower than that in Shanghai. Enrofloxacin, penicillin, and erythromycin were the most detected VAs in raw and cooked food. Only oxytetracycline was detected in terminal tap water, and none were detected in milk. Tetracycline and doxycycline hyclate were detected in organic eggs (up to 7.1ng/g) and regular eggs (up to 6.6ng/g), which were common in children's diets. Traditional Chinese cooking processes did not completely eliminate VAs, and the concentrations of some VAs increased, especially after frying and roasting. The estimated daily intake (EDI) results show that the contribution of dietary intake and that based on the urine concentrations of VAs were far below the acceptable daily intake (ADI). The EDIs from urine were significantly lower than those based on cooked foods. The highest level of achievement percentage (LAP) based on dietary consumption and urine concentrations were 39.7% and 1.79%, respectively, and thus current levels of exposure to VAs would not seem to pose a risk to children's health. However, harmful effects of residual VAs during developmental periods may occur with exposure to much lower doses than those considered harmful to adults, and further investigation of these emerging pollutants is urgently encouraged.

Assessment of Aerobic Plate Counts, <i>Staphylococcus aureus</i>, <i>Escherichia coli</i> and <i>Salmonella</i> in Meat Sold by Street Vendors in the Eastern Cape Province, South Africa

The study was carried out to determine the aerobic plate counts (APC), Staphylococcus aureus, Escherichia coli and Salmonella contamination levels in cooked (n=24) and raw (n=24) beef, pork and mutton samples, surface contact plates (n=48) and water samples (n= 40) from street vendors. A total of 8 street vendors who were willing to participate in the study were randomly selected. After biochemical tests, no significant differences were found in the microbial counts of meat sold by street vendors in Alice and King Williams town. Furthermore, no significant differences were found in the mean scores of raw beef, mutton and pork where APC (4.8, 3.7 and 2.8 Log CFU/g), Staphylococcus aureus (3.3, 3.7 and 2.8 Log CFU/g) and E. coli (1.0, 0.6 and 0.3 Log CFU/g) respectively. Salmonella tested negative in all the samples tested in the study. The results in the study were associated with cross-contamination during processing and storage. However, the levels of contamination in cooked meat were lower when compared to the standards set by Commission Regulation for determining the microbiological quality of ready-to-eat foods. Overall, poor hygiene of the street vendor, utensils, and holding area were major sources of contamination. It was therefore concluded that there were no differences in the microbial counts of meat sold in the informal markets of Nkonkobe and Buffalo City Municipalities in the Eastern Cape Province, South Africa. Improved sanitation facilities, hygiene tools, and training will promote the production of safer food by the street vendors.

AUTENTIKASI DAGING AYAM SEGAR DARI KONTAMINASI DAGING BABI MENGGUNAKAN GEN CYT-B DENGAN ANALISIS DUPLEX- POLYMERASE CHAIN REACTION

Halal is one of important aspects in consumer protection. Meat and processed meat products are food that should be controlled strictly because those are prone to be adulterated by pork contamination. Therefore, it is necessary to provide detection technique which is accurate, fast and cheap. The objective of this research was to identify the presence of impurities of pork meat on raw chicken meat using gene Cyt-b with duplex-PCR analysis. This research used six samples of raw chicken meat and raw pork. Raw chicken meat was bought from supermarkets in the city of Surakarta and raw pork was obtained from pig slaughterhouse. The percentage of raw pork contamination on raw chicken meat was designed as much as 1, 5, 10, and 25%, respectively. The DNA genome was isolated according to DNA isolation protocol from Genomic DNA Mini Kit. In addition, duplex-PCR was performed based on protocol of KAPA2G Fast Multiplex PCR kit. The data was descriptively analyzed by directly looking the DNA bands on the gel documentation apparatus. The result showed that specific DNA bands for chicken and pig were completely appeared on 1.5% of agarose gels. Duplex-PCR detect contamination of pork on raw meat of chicken at all contamination levels. This research proved that the duplex-PCR detect the contamination of pork until the level of 1%.

Lactic acid bacteria isolated from raw and fermented pork products: Identification and characterization of catalase-producing Pediococcus pentosaceus.

Lactic acid bacteria (LAB) from raw and fermented pork samples were screened for their inhibitory activity by an agar spot test in order to obtain a LAB strain with suitable property to be used as meat starter cultures. Among the 174 isolates, 73 were positive to inhibit at least one of the seven indicator bacteria, which were further characterized. The most suitable isolate was isolate P0805, identified as Pediococcus pentosaceus. This bacterium was catalase- and nitrate reductase-positive and amino acid decarboxylase-negative; moreover, it produced inhibitory substances against Salmonella Typhimurium with the activity of the partially purified inhibitory substances of 409,600 AU/mL. To further characterize the catalase-producing ability of P. pentosaceus P0805, the effect of hematin on its catalase activity in Sausage Model Broth (SMB) was evaluated, and it enhanced catalase production. The catalase activity was found in both SMB with and without hematin. It was concluded that catalase produced by this bacterium was heme-independent catalase.

Comparative study of mineral composition of beef steak and pork chops depending on the thermal preparation method

This study focuses on the effects of three different thermal preparation methods (roasting, boiling, and microwave cooking) on the mineral concentrations of beef and pork, as well as on the comparison of mineral levels between these two types of meat. In this study, raw and cooked beef and pork samples were selected and analyzed by ICP-OES in order to determine mineral concentrations. In general, thermal preparation clearly increased mineral concentrations in cooked samples compared to raw meat. The highest mineral concentration was identified in the roasted samples. Trace element concentrations in beef were significantly higher compared to pork. In pork, Na concentration decreased in all samples, suggesting that Na is lost with water. Zn mean content in cooked beef samples registered significant differences compared to pork cooked samples. The percentage of water loss during the microwave thermal preparation for beef samples was higher than the other two treatments.

Effect of low-pressure plasma treatment on the color and oxidative stability of raw pork during refrigerated storage.

The effect of low-pressure plasma on quality attributes of meat is an important aspect, which must be considered before application in food. The aim of this study was to determine the color, fatty acid composition, lipid oxidation expressed as thiobarbituric acid reactive substances and total antioxidant capacity of raw pork samples exposed to helium low-pressure plasma treatment (20 kPa) for 0, 2, 5, and 10 min during the storage period. The thiobarbituric acid reactive substance concentrations of all plasma-treated samples during storage were in the range from 0.26 to 0.61 mg malondialdehyde/kg. Exposure time caused significant changes only in total color difference, hue angle, and chroma after 10 min of treatment. Ferric reducing ability of plasma values of meat samples decreased from 1.93 to 1.40 mmol Trolox Eq/kg after 14 days of storage. The storage period significantly affected proportion of polyunsaturated fatty acids, with an increase about 3% after 14 days of refrigeration storage while the content of saturated fatty acids was at the same level. Helium low-pressure plasma does not induce oxidative processes. Application of this decontamination technique while maintaining product quality is possible in food industry.

Acridine derivatives (PANHs, azaarenes) in raw, fried or grilled pork from different origins, and PANH formation during pork thermal processing

The aim of this work was to determine level of azaarenes (PANHs) in raw pork and to investigate their formation during meat frying or grilling, in particular to verify a suggestion that endogenous vitamin E might inhibit production of azaarenes. Azaarene concentration in raw pork samples from various origins ranged from 2.75ngg−1 to 3.69ngg−1 (2.75–2.93ngg−1 in meat of Polish Landrace pigs, 3.00–3.69ngg–1 in meat of hybrid Duroc×Polish Landrance pigs). PANH formation during frying of pork meat was not confirmed. On the other hand, PANHs were indeed formed during grilling; their levels ranged from 6.21ngg−1 to 8.08ngg–1. No inhibition influence of vitamin E on formation of PANHs on was found either in fried or grilled pork meat.

Vancomycin-resistance phenotypes, vancomycin-resistance genes, and resistance to antibiotics of enterococci isolated from food of animal origin.

In the present study, 500 raw beef, pork, and chicken meat samples and 100 pooled egg samples were analyzed for the presence of vancomycin-resistant enterococci, vancomycin-resistance phenotypes, and resistance genes. Of 141 isolates of enterococci, 88 strains of Enterococcus faecium and 53 strains of E. faecalis were identified. The most prevalent species was E. faecium. Resistance to ampicillin (n = 93, 66%), ciprofloxacin (n = 74, 52.5%), erythromycin (n = 73, 51.8%), penicillin (n = 59, 41.8%) and tetracycline (n = 52, 36.9%) was observed, while 53.2% (n = 75) of the isolates were multiresistant and 15.6% (n = 22) were susceptible to all antibiotics. Resistance to vancomycin was exhibited in 34.1% (n = 30) of the E. faecium isolates (n = 88) and 1.9% (n = 1) of the E. faecalis isolates (n = 53) using the disc-diffusion test and the E-test. All isolates were tested for vanA and vanB using real-time polymerase chain reaction (PCR) and multiplex PCR, and for vanC, vanD, vanE, vanG genes using multiplex PCR only. Among E. faecalis isolates, no resistance genes were identified. Among the E. faecium isolates, 28 carried the vanA gene when tested by multiplex PCR and 29 when tested with real-time PCR. No isolate carrying the vanC, vanD, vanE, or vanG genes was identified. Melting-curve analysis of the positive real-time PCR E. faecium isolates showed that 22 isolates carried the vanA gene only, 2 isolates the vanB2,3 genes only, and seven isolates carried both the vanA and vanB2,3 genes. Enterococci should be considered a significant zoonotic pathogen and a possible reservoir of genes encoding resistance potentially transferred to other bacterial species.

SCCmec Type IX Element in Methicillin Resistant Staphylococcusaureusspa Type t337 (CC9) Isolated from Pigs and Pork in Thailand

Methicillin resistant Staphylococcus aureus (MRSA) have emerged among livestock in several countries. In this study, we describe the results of a screening performed in pigs and raw pork samples in Thailand. Ten pork samples and 15 nasal swabs from pigs were collected from 2 markets and 1 pig farm in the Samuth Songkhram province in Thailand. MRSA were isolated using selective isolation procedures and confirmed by mecA PCR. The MRSA were characterized by antimicrobial susceptibility testing, pulsed field gel electrophoresis (PFGE), spa typing, SCCmec typing, and MLST. Resistance and virulence markers were screened using a microarray. Five of the pork samples and six pig nasal swabs were positive for MRSA. All 11 isolates belonged to spa type t337 but showed diversity in antimicrobial resistance patterns and PFGE profiles. Additionally, the isolates were sequence-typed; ST9, ST2136, ST2278 belonging to the clonal complex; CC9. All isolates harbored SCCmec IX and were resistant to 7 out of 14 tested antimicrobials; additional resistances to all antimicrobials tested were found in some of the pork and pig isolates and 1 pork isolate was resistant to 13 antimicrobials tested. Microarray analysis identified blaZ, aac-aphD, vga(A), tetM, and a tet efflux marker, in all strains and additionally ermB and aadD, cat and fex(A) in the pork isolates. None of the isolates were found PVL-positive, but enterotoxins were identified in all isolates. To our knowledge, only a few descriptions of MRSA in livestock and food products in Thailand have been observed but this is the first observation of MRSA CC9 associated with SCCmec IX in pork. This study indicates a likely widespread distribution of MRSA in pig and pork in Thailand and further investigation on the prevalence and importance of livestock associated MRSA in Thailand is needed.

Prevalence of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) on retail meat in Iowa

Several recent studies have indicated a high prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in retail-available meat. However, few studies have investigated MRSA in meat in the United States. The aim of this study was to determine the presence of Staphylococcus aureus (S. aureus) on meat samples available at retail stores. Samples of fresh raw pork, chicken, beef, and turkey were purchased from 22 food stores throughout Iowa. S. aureus strains were isolated from 27 of 165 samples, giving an overall prevalence of 16.4%. Turkey, pork, chicken, and beef had individual S. aureus prevalence rates of 19.4%, 18.2%, 17.8%, and 6.9%, respectively. Two isolates of MRSA were isolated from pork, giving an overall prevalence of 1.2%. One MRSA isolate was positive for the PVL gene. Common spa types included t034, t337, t008, and t002. These results suggest that MRSA is present on low numbers of retail meat in Iowa.

Identification and Molecular Characterization of Antimicrobial-Resistant Shiga Toxin–ProducingEscherichia coliIsolated from Retail Meat Products

Ten (2.7%) Shiga toxin-producing Escherichia coli (STEC) were isolated from 370 samples of raw minced beef, mutton, pork, and chicken from the Jilin region of China; and additional 10 E. coli O157:H7 isolates were previously isolated from different Jilin regions. Seventeen of the isolates were multiresistant, exhibiting resistance to ampicillin, ciprofloxacin, tetracycline, sulfamethoxazole-trimethoprim, gentamycin, and streptomycin. Class 1 integrons were detected in nine (45.0%) of the STEC isolates and consisted of serogroups O157, O62, O113, O149, and O70. Integrons containing amplicons of a 0.5-1.5 or 1.0 kb gene cassette were found in seven (77.8%) of the integron-containing isolates. Sequencing analysis revealed that these gene cassettes encode genes conferring resistance to trimethoprim (dfrA1) and streptomycin (aadA1). The 0.5 kb cassette described here was found to encode a putative transporter peptide in the STEC. Seventeen isolates contained plasmids with different bands, and transfer by conjugation between strains of E. coli demonstrated that class 1 integrons located on mobile plasmids could contribute to the emergence and dissemination of antimicrobial resistance to ampicillin, gentamycin, streptomycin, and sulfamethoxazole-trimethoprim amongst STEC. These data revealed the high prevalence of antimicrobial-resistant STEC isolates in Jilin's surrounding regions, providing important and useful surveillance information reflecting antimicrobial selection pressure.

Prevalence of methicillin-resistant Staphylococcus aureus in meat

Recently the isolation of methicillin-resistant Staphylococcus aureus (MRSA) strains from several food-producing animals has been reported. During slaughtering of MRSA-positive animals, contamination of carcasses with MRSA may occur and consequently the meat of these animals may get contaminated. The aim of this study was to estimate the prevalence of MRSA in raw meat samples from the retail trade. Samples of raw beef, pork, veal, lamb/mutton, chicken, turkey, fowl and game were collected from the retail trade. A detection method including a two-step enrichment in Mueller–Hinton broth + 6.5% NaCl and phenol red mannitol broth containing ceftizoxime and aztreonam, followed by isolation on MRSA ID agar (bioMérieux) was evaluated and subsequently applied for the detection of MRSA in samples of raw meats. MRSA strains were isolated from 264 (11.9%) of 2217 samples analyzed. Isolation percentages for the meat species were: beef (10.6%), veal (15.2%), lamb and mutton (6.2%), pork (10.7%), chicken (16.0%), turkey (35.3%), fowl (3.4%) and game (2.2%). The majority (85%) of the isolated strains belonged to spa-types of pulsed-field gel electrophoresis (PFGE) non-typeable (NT)-MRSA, corresponding to the multilocus sequence type ST398, a type also recently isolated in the Netherlands from pigs. However, a smaller part of these strains were found to be of other ST's, possibly of human origin. Further studies are needed to elucidate transmission routes of MRSA in relation to meat and other foods and to provide the tools for preventing the spread of MRSA. At present the high prevalence of MRSA in meat has not been shown to contribute significantly to the dissemination of MRSA to humans and the possible health hazard for consumers of the presence of MRSA in foods should be further elucidated.

한국형 육포제조를 위한 원료 돈육의 미생물 분포 및 병원성 미생물의 확인

시중의 정육점 및 백화점 등에서 유통 중인 10종의 돈육 원료에 대한 일반세균, 저온균, 고온균, 혐기성균 및 진균류, 대장균군 등의 미생물학적 분포와 병원성 미생물에 대한 분리동정을 실시하였다. 실험 결과, 원료 돈육에서 중온균은 <TEX>$3.9{\times}10^2-3.9{\times}10^5cfu/g$</TEX>으로 높은 분포를 보였고, 저온균은 <TEX>$1.5{\times}10^3-8.6{\times}10^2cfu/g$</TEX>, 혐기성균은 중온균, 저온균과 유사한 분포를 보였으나 상대적으로 적게 검출되었고, 고온균은 모든 검체에서 검출되지 않았다. 대장균군 또한 모든 검체에 대해서 검출되지 않았으며, 곰팡이와 효모 등 진균류는 <TEX>$3.8{\times}10^1-5.1{\times}10^2cfu/g$</TEX>으로 검출되었다. B. cereus는 돈육 sample B와 J에서 분리되었고, S. aureus의 경우 돈육 sample B에서만 검출되었다. B. cereus는 API kit를 이용한 간이동정에서 99.8%의 상동성을 보였고, S. aureus는 97.8%의 상동성을 보였다. The objective of this study is to evaluate the microbial safety of raw pork used to produce Korean pork jerky. The raw pork samples harbored large populations of microorganisms. In particular, mesophilic bacteria were found to be most numerous <TEX>$(3.9{\times}10^2-3.9{\times}10^5cfu/g)$</TEX> in the samples. Spore-forming bacteria and coliforms were not detected below detection limit. Yeast and molds were detected at <TEX>$3.8{\times}10^1-5.1{\times}10^2cfu/g$</TEX> in the raw pork. Ten samples of raw pork were analyzed for the presence of pathogenic bacteria. Bacillus cereus was isolated from samples B and J and Staphylococcus aureus was isolated from sample B. The B. cereus isolates from raw pork samples were identified with 99.8% agreement and S. aureus isolate was identified with 97.8% agreement according to the API CHB 50 kit.

Predicting Pathogen Growth during Short-Term Temperature Abuse of Raw Pork, Beef, and Poultry Products: Use of an Isothermal-Based Predictive Tool

A computer-based tool (available at: www.wisc.edu/foodsafety/meatresearch) was developed for predicting pathogen growth in raw pork, beef, and poultry meat. The tool, THERM (temperature history evaluation for raw meats), predicts the growth of pathogens in pork and beef (Escherichia coli O157:H7, Salmonella serovars, and Staphylococcus aureus) and on poultry (Salmonella serovars and S. aureus) during short-term temperature abuse. The model was developed as follows: 25-g samples of raw ground pork, beef, and turkey were inoculated with a five-strain cocktail of the target pathogen(s) and held at isothermal temperatures from 10 to 43.3°C. Log CFU per sample data were obtained for each pathogen and used to determine lag-phase duration (LPD) and growth rate (GR) by DMFit software. The LPD and GR were used to develop the THERM predictive tool, into which chronological time and temperature data for raw meat processing and storage are entered. The THERM tool then predicts a Δ log CFU value for the desired pathogen-product combination. The accuracy of THERM was tested in 20 different inoculation experiments that involved multiple products (coarse-ground beef, skinless chicken breast meat, turkey scapula meat, and ground turkey) and temperature-abuse scenarios. With the time-temperature data from each experiment, THERM accurately predicted the pathogen growth and no growth (with growth defined as Δ log CFU ≥ 0.3) in 67, 85, and 95% of the experiments with E. coli O157:H7, Salmonella serovars, and S. aureus, respectively, and yielded fail-safe predictions in the remaining experiments. We conclude that THERM is a useful tool for qualitatively predicting pathogen behavior (growth and no growth) in raw meats. Potential applications include evaluating process deviations and critical limits under the HACCP (hazard analysis critical control point) system.

Detection of Pathogenic Yersinia enterocolitica in Meat using Real-Time PCR

Yersinia enterocolitica is an important zoonosis, which can cause disease in humans and animals. The culture methods available for detection of Y. enterocolitica in food samples are time-consuming and seldom successful. Using DNA-based methods, like PCR, this pathogen can be detected more rapidly and with greater sensitivity. The aim of this study was to establish a rapid and accurate real-time PCR method to detect pathogenic Y. enterocolitica in pork. The chromosomal ail−gene, which is only present in pathogenic Y. enterocolitica strains, was used as DNA target for the 5' nuclease PCR assay. The probe was labelled at the 5' end with the fluorescent reporter dye (FAM) and at the 3' end with the quencher dye (TAMRA). A 2-step protocol with 45 cycles was used in a multicolour real-time PCR detection system. A Ct value over 40 indicated a negative result. The DNA extraction procedure for the natural samples was rapid and simply. This qualitative real-time PCR method was shown to be specific and sensitive. Detection rate of ail-positive Y. enterocolitica in 200 pig tonsils was 88 % and 35 % with PCR and culture methods, respectively. When 100 raw pork samples were studied, 7 were positive with PCR and all were culture negative.

Evaluation of a Combined Culture and PCR Method (NMKL-163A) for Detection of Presumptive Pathogenic Yersinia enterocolitica in Pork Products

A combined culture and PCR method for detection of pathogenic Yersinia enterocolitica in food (NMKL-163A) was evaluated by testing samples of artificially and naturally contaminated pork. The performance of the pre-PCR sample treatment, buoyant density centrifugation, was first compared with two commercially available methods (DNeasy tissue kit and PrepMan). We found that similar sensitivity was reached (i.e., 25 CFU/g of food was detected by single PCR) with the buoyant density centrifugation and the DNeasy Tissue kit when tested on overnight enrichments. However, the DNeasy tissue kit was superior when tested on nonenriched homogenates; the detection limit was 25 CFU/g in minced beef by single PCR and 25 CFU/g in sausage by nested PCR. We then analyzed 100 raw minced pork samples. Thirty-five tested positive for presumptive pathogenic Y. enterocolitica when analyzed by the NMKL-163A method, whereas none tested positive when analyzed in parallel by a standard culture method (ISO 10273). We also analyzed 97 samples of cold-smoked pork sausage, of which approximately 11% tested positive by the NMKL-163A method. This study showed that sensitivities such as those obtained by nested PCR were required for detection of the pathogen in naturally contaminated samples, and therefore the nested PCR primers, which are included in the NMKL-163A method only as an option, need to be validated and applied routinely.

Prevalence of Arcobacter spp. in Raw Milk and Retail Raw Meats in Northern Ireland

A 1-year study was undertaken to determine the prevalence of Arcobacter spp. in raw milk and retail raw meats on sale in Northern Ireland. Retail raw poultry samples (n = 94), pork samples (n = 101), and beef samples (n = 108) were obtained from supermarkets in Northern Ireland, and raw milk samples (n = 101) were kindly provided by the Milk Research Laboratory, Department of Agriculture and Rural Development, Belfast, Northern Ireland. Presumptive arcobacters were identified by previously described genus-specific and species-specific PCR assays. Arcobacter spp. were found to be common contaminants of retail raw meats and raw milk in Northern Ireland. Poultry meat (62%) had the highest prevalence, but frequent isolations were made from pork (35%), beef (34%), and raw milk (46%). Arcobacter butzleri was the predominant species isolated from retail raw meats and was the only species isolated from raw milk samples. Arcobacter cryaerophilus was detected less frequently, and Arcobacter skirrowii was detected only as a cocontaminant. To our knowledge, this is the first report of Arcobacter spp. prevalence in a diverse range of products of animal origin in Northern Ireland.

Identification and Characterization of Pathogenic Yersinia enterocolitica Isolates by PCR and Pulsed-Field Gel Electrophoresis

Approximately 550 to 600 yersiniosis patients are reported annually in Sweden. Although pigs are thought to be the main reservoir of food-borne pathogenic Yersinia enterocolitica, the role of pork meat as a vehicle for transmission to humans is still unclear. Pork meat collected from refrigerators and local shops frequented by yersiniosis patients (n=48) were examined for the presence of pathogenic Yersinia spp. A combined culture and PCR method was used for detection, and a multiplex PCR was developed and evaluated as a tool for efficient identification of pathogenic food and patient isolates. The results obtained with the multiplex PCR were compared to phenotypic test results and confirmed by pulsed-field gel electrophoresis (PFGE). In all, 118 pork products (91 raw and 27 ready-to-eat) were collected. Pathogenic Yersinia spp. were detected by PCR in 10% (9 of 91) of the raw pork samples (loin of pork, fillet of pork, pork chop, ham, and minced meat) but in none of the ready-to-eat products. Isolates of Y. enterocolitica bioserotype 4/O:3 were recovered from six of the PCR-positive raw pork samples; all harbored the virulence plasmid. All isolates were recovered from food collected in shops and, thus, none were from the patients' home. When subjected to PFGE, the six isolates displayed four different NotI profiles. The same four NotI profiles were also present among isolates recovered from the yersiniosis patients. The application of a multiplex PCR was shown to be an efficient tool for identification of pathogenic Y. enterocolitica isolates in naturally contaminated raw pork.

Specific detection of cytopathogenic Listeria monocytogenes using a two-step method of immunoseparation and cytotoxicity analysis

The development of rapid methods for detection of viable Listeria monocytogenes is crucial to prevent listeriosis and product recalls. While immunomagnetic separation has been used for isolating Listeria spp., lack of specificity and pathogenicity determination render this method unsatisfactory. A two-step method using Protein A agarose beads (Immunobeads) coated with a more specific antibody, monoclonal antibody (MAb)-C11E9 for L. monocytogenes was developed. Immunobeads were allowed to capture Listeria cells from a variety of samples and tested for cytopathogenic action on a murine hybridoma B-lymphocyte, Ped-2E9 cell line by Trypan blue staining, and by an alkaline phosphatase (AP)-based cytotoxicity assay. The two-step method was used to test uninoculated hotdogs, bologna, and raw beef, chicken, and pork samples, following selective enrichment in half-Fraser broth. Pure culture studies proved the assay to be specific for L. monocytogenes, while a similar assay with Dynal® Anti-Listeria immunomagnetic beads was positive for L. monocytogenes, L. ivanovii, and L. seeligeri. Detection and confirmation of cytopathogenicity of Listeria cells from food samples after 24-h selective enrichment were completed in 2–4 h. Isolates were further analyzed by the CAMP test for hemolytic activity and RiboPrinter® for genomic patterns. Using immunoseparation and cytotoxicity as a two-step rapid method, viable L. monocytogenes could be isolated, detected, and confirmed as cytopathogenic in 28 h or less.