Fecal microbiota composition, serum metabolomics, and markers of inflammation in dogs fed a raw meat-based diet compared to those on a kibble diet.

Introduction: Despite the potential health risks associated with feeding raw and non-traditional diets, the use of these diets in dogs is increasing, yet the health outcomes associated with these diets is not well understood. This study investigates the effect of feeding dogs a kibble or raw meat-based diets on fecal microbiota composition, serum metabolomics and inflammatory markers. Methods: Clinically healthy dogs with a history of consuming either kibble (KD, n = 27) or raw meat-based diets (RMBD, n = 28) for more than 1 year were enrolled. Dogs were fed a standardized diet of either a single brand of KD or RMBD for 28 days. Serum and fecal samples were collected for analysis of microbiota, metabolomics, and inflammatory markers. Multiple regression analysis was performed for each of the metabolites and inflammatory markers, with feed group, age and BCS included as independent variables. Results: The fecal microbiota composition differed between the KD and RMBD groups. Beta-diversity and some indices of alpha-diversity (i.e., Shannon and Simpson) were different between the two diet groups. Sixty- three serum metabolites differed between KD and RMBD-fed dogs with the majority reflecting the differences in macronutrient composition of the two diets.Fecal IAP, IgG and IgA were significantly higher in RMBD dogs compared to KD dogs, while systemic markers of inflammation, including serum c-reactive protein (CRP), galectin, secretory receptor of advanced glycation end-products (sRAGE), haptoglobin, and serum IgG were similar in dogs fed either diet. Discussion: Diet composition significantly affected fecal microbiota composition and metabolome. Although it had a potentially beneficial effect on local inflammatory markers, feeding RMBD had no impact on systemic inflammation. The influence of these changes on long term health outcomes provides an area for future study.

Raw meat-based diet for pets: a neglected source of human exposure to Salmonella and pathogenic Escherichia coli clones carrying mcr, Portugal, September 2019 to January 2020.

BackgroundThe pet industry is expanding worldwide, particularly raw meat-based diets (RMBDs). There are concerns regarding the safety of RMBDs, especially their potential to spread clinically relevant antibiotic-resistant bacteria or zoonotic pathogens.AimWe aimed to investigate whether dog food, including RMBD, commercially available in Portugal can be a source of Salmonella and/or other Enterobacteriaceae strains resistant to last-line antibiotics such as colistin.MethodsFifty-five samples from 25 brands (21 international ones) of various dog food types from 12 suppliers were screened by standard cultural methods between September 2019 and January 2020. Isolates were characterised by phenotypic and genotypic methods, including whole genome sequencing and comparative genomics.ResultsOnly RMBD batches were contaminated, with 10 of 14 containing polyclonal multidrug-resistant (MDR) Escherichia coli and one MDR Salmonella. One turkey-based sample contained MDR Salmonella serotype 1,4,[5],12:i:- ST34/cgST142761 with similarity to human clinical isolates occurring worldwide. This Salmonella exhibited typical antibiotic resistance (bla TEM + strA-strB + sul2 + tet(B)) and metal tolerance profiles (pco + sil + ars) associated with the European epidemic clone. Two samples (turkey/veal) carried globally dispersed MDR E. coli (ST3997-complexST10/cgST95899 and ST297/cgST138377) with colistin resistance (minimum inhibitory concentration: 4 mg/L) and mcr-1 gene on IncX4 plasmids, which were identical to other IncX4 circulating worldwide.ConclusionSome RMBDs from European brands available in Portugal can be a vehicle for clinically relevant MDR Salmonella and pathogenic E. coli clones carrying genes encoding resistance to the last-line antibiotic colistin. Proactive actions within the One Health context, spanning regulatory, pet-food industry and consumer levels, are needed to mitigate these public health risks.

Development of a closed-tube, calcein-based loop-mediated isothermal amplification assay to detect Salmonella spp. in raw meat samples.

Foodborne pathogens compromise food safety and public health, and Salmonella spp. are among the major pathogenic bacteria that cause outbreaks worldwide. Proper surveillance through timely and cost-effective detection methods across the food animal production chain is crucial to prevent Salmonella outbreaks and agricultural losses. Traditional culture methods are labor- and resource-intensive, with lengthy turnaround times. Meanwhile, conventional molecular tools, such as PCR and qPCR, are expensive and require technical skills and equipment. Loop-mediated isothermal amplification (LAMP) is a simple, rapid, inexpensive, highly sensitive, and specific molecular assay that does not require expensive equipment. Hence, this study developed and optimized a closed-tube, calcein-based LAMP assay to detect Salmonella using the invA gene and performed evaluation and validation against conventional PCR. The LAMP assay showed high specificity and sensitivity. It showed 10-fold higher sensitivity than conventional PCR, at <1 ng/µL DNA concentrations. Meanwhile, for CFU/mL, LAMP assay showed 1000-fold higher sensitivity than conventional PCR at 4.8 × 103 cells/mL than 4.8 × 107 cells/mL, respectively. For parallel testing of 341 raw meat samples, after conventional culture enrichment (until Rappaport-Vassiliadis broth), the optimized LAMP assay showed 100% detection on all samples while conventional PCR showed 100%, 99.04%, and 96.64% for raw chicken, beef, and pork samples, respectively. Meanwhile, a shortened enrichment protocol involving 3-h incubation in buffered peptone water only, showed lower accuracy in tandem with the optimized LAMP assay ranging from 55 to 75% positivity rates among samples. These suggest that the optimized LAMP assay possesses higher sensitivity over conventional PCR for invA gene detection when coupled with conventional enrichment culture methods. Hence, this assay has potential as a powerful complementary or alternative Salmonella detection method to increase surveillance capacity and protect consumer food safety and public health worldwide.

Assessment of bacteriological quality and safety of raw meat at slaughterhouse and butchers' shop (retail outlets) in Assosa Town, Beneshangul Gumuz Regional State, Western Ethiopia.

Contaminated meat has been implicated in many cases of foodborne illness and poses serious challenges in developing countries. This study aimed to assess the quality and safety of raw beef meat in Assosa Town. The finding showed that the mean of Aerobic mesophilic bacteria (AMB) and S. aureus at retail outlets was 5.04 log10cfu/g and 3.84 log10cfu/g; 4.03 log10cfu/g and 3.5 log10cfu/g at slaughterhouse, respectively. The microbial load range of AMB at the butcher shop was 2.49-5.16 log10 cfu/g, while at the abattoir it was 2.75-7.52 log10 cfu/g out of 70 raw beef meat analyzed samples. Similar to this, the butcher shop and abattoir had S. aureus microbiological load ranges of 2.74 - 4.84 log10 cfu/g and 2.71-4.72 log10 cfu/g, respectively. In contrast, 25.7% and 34.3% of the samples in the abattoir and retail shop, respectively, were contaminated with Salmonella sp. For S. aureus, just 38.71% and 17.14%, respectively, of the samples at the retail and butcher shops were satisfactory. AMB found that 80% of the examined samples from butcher shops and 57.7% from abattoirs were satisfactory. Due to poor handling and environmental hygiene procedures by Assosa Town butchers, 77.1% of the meat contact surface and 82.9% of the carcass were exposed to flies. On the other hand, only 5.7%, 28.6%, and 22.9% of the butchers kept the carcass in the refrigerator, and wore gowns and hairnets, respectively. In slaughterhouses, the majority of respondents (87.5%) concur that there were certain challenges in achieving slaughtering in the working environment.

Detection of virulence genes of Staphylococcus aureus isolated from raw beef for retail sale in the markets of Ulaanbaatar city, Mongolia.

BACKGROUND: Staphylococcus aureus (S. aureus) is a highly virulent pathogen that causes food-borne illness, food poisoning, skin and soft tissue infections, abscesses, mastitis, and bacteremia. It is common for meat and meat products to become contaminated with S. aureus due to dirty hands, food storage conditions, food production processes, and unhygienic conditions, causing food poisoning. Therefore, we aimed to isolate S. aureus strain from the raw beef and reveal virulence genes and antibiotic resistance profile from isolated S. aureus strains. METHODS: In this study, 100 samples of raw beef were collected from 4 major market stalls in Ulaanbaatar city, Mongolia. S. aureus was detected according to the ISO 6888-1:2021 standard, and the nucA gene encoding the species-specific thermonuclease was amplified and confirmed by polymerase chain reaction (PCR). In the strains of S. aureus isolated from the samples, the genes encoding the virulence factors including sea, sed, tsst, eta, etb, and mecA were amplified by multiplex PCR. These genes are encoded staphylococcal enterotoxin A, enterotoxin D, toxic shock syndrome toxin, exotoxin A, exotoxin B and penicillin-binding protein PBP 2A, respectively. Antibiotic sensitivity test was performed by the Kirby-Bauer disc diffusion method. The Clinical and Laboratory Standard Institute guidelines as CLSI M100-S27 was used for analysis of the data. RESULTS: Thirty-five percent of our samples were detected contaminated with of the S. aureus strains. Subsequently, antibiotic resistance was observed in the S. aureus contaminated samples. Among our samples, the highest rates of resistance were determined against ampicillin (97.1%), oxacillin (88.6%), and penicillin (88.6%), respectively. Three genes including mecA, sea, and tsst from six virulence genes were detected in 17% of S. aureus strain-contaminated samples by multiplex PCR. The sed, etb and eta genes were detected in the 2.9%, 11.4% and 5.7% of our samples, respectively. CONCLUSION: The results show that S. aureus related contamination is high in the raw beef for retail sale and prevalent S. aureus strains are resistant to all antibiotics used. Also, our results have demonstrated that there is a high risk for food poisoning caused by antibiotic resistant S. aureus in the raw beef and it may establish public health issues. Genes encoding for both heat-resistant and nonresistant toxicity factors were detected in the antibiotic resistant S. aureus strains and shown the highly pathogenic. Finally, our study is ensuring to need proper hygienic conditions during beef's preparation and sale.

Antimicrobial-Resistant Escherichia coli, Enterobacter cloacae, Enterococcus faecium, and Salmonella Kentucky Harboring Aminoglycoside and Beta-Lactam Resistance Genes in Raw Meat-Based Dog Diets, USA.

The practice of feeding raw meat-based diets to dogs has grown in popularity worldwide in recent years. However, there are public health risks in handling and feeding raw meat-based dog diets (RMDDs) to dogs since there are no pathogen reduction steps to reduce the microbial load, which may include antimicrobial-resistant pathogenic bacteria. A total of 100 RMDDs from 63 suppliers were sampled, and selective media were used to isolate bacteria from the diets. Bacterial identification, antimicrobial susceptibility testing, and whole-genome sequencing (WGS) were conducted to identify antimicrobial resistance (AMR). The primary meat sources for RMDDs included in this study were poultry (37%) and beef (24%). Frozen-dry was the main method of product production (68%). In total, 52 true and opportunistic pathogens, including Enterobacterales (mainly Escherichia coli, Enterobacter cloacae) and Enterococcus faecium, were obtained from 30 RMDDs. Resistance was identified to 19 of 28 antimicrobials tested, including amoxicillin/clavulanic acid (23/52, 44%), ampicillin (19/52, 37%), cephalexin (16/52, 31%), tetracycline (7/52, 13%), marbofloxacin (7/52, 13%), and cefazolin (6/52, 12%). All 19 bacterial isolates submitted for WGS harbored at least one type of AMR gene. The identified AMR genes were found to mediate resistance to aminoglycoside (gentamicin, streptomycin, amikacin/kanamycin, gentamicin/kanamycin/tobramycin), macrolide, beta-lactam (carbapenem, cephalosporin), tetracycline, fosfomycin, quinolone, phenicol/quinolone, and sulfonamide. In conclusion, the results of this study suggest that feeding and handling RMDDs may pose a significant public health risk due to the presence of antimicrobial-resistant pathogens, and further research and intervention may be necessary to minimize these risks.

Brucella suis biovar 1 infection in a dog with orchitis in Germany.

In 2021, a case of canine brucellosis diagnosed in a dog with orchitis was presented to a veterinary practice in Germany. Serological testing excluded Brucella (B.) canis as a causative agent, but molecular analysis revealed the presence of B. suis biovar 1. Since biovar 1 is not endemic in Europe and the dog had no history of travel to endemic areas, a comprehensive epidemiological investigation was conducted using whole genome sequence data to determine the source of infection. We describe the clinical progress of the animal and the potential infection of a veterinary clinic employee. The findings highlight the importance of considering less common Brucella species as possible causes of canine brucellosis. The data also emphasize that it is quite challenging to identify Brucella species in a routine diagnostic laboratory and to conduct epidemiological investigations to unveil possible transmission routes.

Bacterial Pericarditis Caused by Campylobacter fetus subsp. fetus After Mutton Consumption.

Campylobacter fetus subsp. fetus causes systemic diseases including bacteremia and meningitis. However, it rarely causes bacterial pericarditis. We present a rare case of bacterial pericarditis caused by Campylobacter fetus subsp. fetus. A man in his 60s presented with a fever and dyspnea. Electrocardiography revealed ST segment elevation in all leads except augmented vector right (aVR), and contrast-enhanced computed tomography of the chest revealed a large pericardial effusion. Campylobacter fetus subsp. fetus, appearing as curved, gull-wing-shaped gram-negative rods on microscopy, was identified on blood culture. The patient was diagnosed with acute pericarditis caused by Campylobacter fetus subsp. fetus. Further, history-taking revealed that he had consumed undercooked mutton before the onset of his illness. He recovered after treatment with antibiotics (ceftriaxone, ampicillin, and amoxicillin) for four weeks. With a blood culture revealing gull-wing shaped gram-negative rods, and the patient's history including potential contact with animals or the consumption of raw or undercooked meat, Campylobacter fetus subsp. fetus infection should be suspected.

Assessment of virulence factors and antimicrobial resistance among the Pseudomonas aeruginosa strains isolated from animal meat and carcass samples.

BACKGROUND: Pseudomonas aeruginosa bacteria are emerging causes of food spoilage and foodborne diseases. Raw meat of animal species may consider a reservoir of P. aeruginosa strains. OBJECTIVES: The present survey was done to assess the prevalence, antibiotic resistance properties and distribution of virulence factors among the P. aeruginosa strains isolated from raw meat and carcass surface swab samples of animal species. METHODS: Five hundred and fifty raw meat and carcass surface swab samples were collected from cattle and sheep species referred to as slaughterhouses. P. aeruginosa bacteria were identified using culture and biochemical tests. The pattern of antibiotic resistance was determined by disk diffusion. The distribution of virulence and antibiotic resistance genes was determined using polymerase chain reaction. RESULTS: Forty-seven of 550 (8.54%) examined samples were contaminated with P. aeruginosa. The prevalence of P. aeruginosa in raw meat and carcass surface swab samples were 6.57 and 12%, respectively. P. aeruginosa isolates showed the maximum resistance rate toward penicillin (87.23%), ampicillin (85.10%), tetracycline (85.10%), gentamicin (65.95%) and trimethoprim (57.44%). The most commonly detected antibiotic resistance genes were BlaCTX-M (53.19%), blaDHA (42.55%) and blaTEM (27.65%). The most commonly detected virulence factors was ExoS (42.55%), algD (31.91%), lasA (31.91%), plcH (31.91%) and exoU (25.53%). CONCLUSIONS: Meat and carcass surface swab samples may be sources of resistant and virulent P. aeruginosa, which pose a hygienic threat in their consumption. However, further investigations are required to identify additional epidemiological features of P. aeruginosa in meat and carcass surface samples.

Clarithromycin-, and metronidazole-resistant Helicobacter pylori isolated from raw and ready-to-eat meat in Mansoura, Egypt.

Helicobacter pylori (H. pylori), classified as a class-I carcinogen, is one of the leading medical pathogens of global concern associated mainly with the development of gastric adenocarcinomas and gastric mucosa-associated lymphoid tissue lymphomas; nevertheless, its prevalence in food especially meat and meat products is not fully covered. Additionally, the resistance of H. pylori towards clarithromycin is increasing worldwide and consider the leading cause of H. pylori treatment failure. Thus, the present study was designed to determine the prevalence, molecular characterization, and antimicrobial resistance profiles of clarithromycin-, and metronidazole-resistant H. pylori isolated from raw and ready-to-eat meat samples retailed in Mansoura city, Egypt. Among the 250 samples tested, H. pylori were molecularly confirmed in 40.8 % (49/120) of raw meat products and in 29.2 % (38/130) of ready-to-eat meat products. Precisely, 53.3 % (32/60), 56.7 % (17/30), 40 % (8/20), 55 % (11/20), 60 % (12/20), 13.3 % (4/30), and 15 % (3/20) of raw ground beef, beef burger, beef burger sandwiches, beef shawarma sandwiches, beef kofta sandwiches, beef luncheon, and beef sausage sandwiches, respectively were positive for H. pylori. Of the 204 biochemically-identified H. pylori isolates, 53.9 % (110/204) were molecularly confirmed by PCR through the detection of glmM, cagA, or vacA genes, which were detected at an incidence of 95.5 % (105/110), 77.3 % (85/110), and 20.9 % (23/110) among the isolates, respectively. The antimicrobial sensitivity testing revealed that all of the 110 (100 %) molecularly-confirmed H. pylori isolates were multidrug-resistant (MDR; resistant to four or more antibiotics). Interestingly, 100 % and 61.8 % of H. pylori isolated from raw and ready-to-eat meat were resistant to metronidazole and clarithromycin, respectively which consider alarming results as metronidazole and clarithromycin are the mainstay antibiotics in the treatment of H. pylori infections. Additionally, 94.5 %, 94.5 %, 24.5 %, 23.6 %, and 13.6 % of isolates were resistant to vancomycin, sulphamethoxazole-trimethoprim, imipenem, levofloxacin, and nitrofurantoin, respectively. The widespread contamination of examined raw and ready-to-eat meat product samples with MDR H. pylori isolates could constitute a tremendous public health hazard. Further studies concerning the prevalence and possible methods of elimination of H. pylori in different food categories distributed in the various provinces in Egypt as well as in other countries is required for a better understanding the H. pylori as an emerging foodborne pathogen.

Raw Meat Consumption and Food Safety Challenges: A Survey of Knowledge, Attitudes, and Practices of Consumers in Lebanon.

A key contributor to foodborne illnesses is consuming contaminated ready-to-eat foods, including raw meats. The latter is a common practice in Lebanon, a country that suffers from widespread pollution and food safety challenges. However, studies on the safety of raw meat consumption in Lebanon are limited. In this study, an attempt was made to investigate the knowledge, attitudes, and practices (KAPs) of the Lebanese population toward the potential risk associated with the consumption of raw meats, and to identify factors that affect KAP levels. An online survey (n = 577) was administered to Lebanese adults aged 18 years and above to assess their KAPs. The results showed that 74.5% of the participants consumed raw meat, 44% had good food safety knowledge, and 30.7% exhibited good practices. However, more than half of the participants (61.9%) showed a positive attitude toward food safety. There was a significant association between knowledge and attitude (p < 0.001), attitude and practices (p < 0.001), and knowledge and practices (p < 0.001), thereby indicating that an increase in food safety education could translate into better practices in this population. Accordingly, efforts to enhance education on food safety are warranted to reduce the potential risk of food poisoning associated with raw meat consumption in Lebanon.

Enterococcus species: insights into antimicrobial resistance and whole-genome features of isolates recovered from livestock and raw meat in Ghana.

Introduction: Enterococcus spp. have gradually evolved from commensals to causing life-threatening hospital-acquired infections globally due to their inherent antimicrobial resistance ability and virulence potential. Enterococcus spp. recovered from livestock and raw meat samples were characterized using antimicrobial susceptibility testing and whole-genome sequencing. Materials and methods: Isolates were confirmed using the MALDI-ToF mass spectrometer, and antimicrobial susceptibility was determined using the Kirby-Bauer disk diffusion method. Whole genome sequencing was performed on isolates resistant to two or more antibiotics. Bioinformatics analysis was performed to determine sequence types, resistance and virulence gene content and evolutionary relationships between isolates from meat and livestock samples, and other enterococci genomes curated by PATRIC. eBURST analysis was used to assign genomes to clonal complexes. Results: Enterococcus spp. were predominantly E. faecalis (96/236; 41%) and E. faecium (89/236; 38%). Overall, isolates showed resistance to erythromycin (78/236; 33%), tetracycline (71/236; 30%), ciprofloxacin (20/236; 8%), chloramphenicol (12/236; 5%), linezolid (7/236; 3%), ampicillin (4/236; 2%) and vancomycin (1/236, 0.4%). Resistance to two or more antimicrobial agents was detected among 17% (n = 40) Enterococcus spp. Resistance genes for streptogramins [lsa(A), lsa(E), msr(C)], aminoglycosides [aac(6')-Ii, aph(3')-III, ant(6)-Ia, aac(6')-aph(2″), str], amphenicol [cat], macrolides [erm(B), erm(T), msr(C)], tetracyclines [tet(M), tet(L), tet(S)] and lincosamides [lsa(A), lsa(E), lnu(B)] were detected among the isolates. Genes for biofilm formation, adhesins, sex pheromones, cytolysins, hyaluronidase, oxidative stress resistance, quorum-sensing and anti-phagocytic activity were also identified. Potential plasmids with replicon sequences (rep1, rep2, repUS43, repUS47, rep9a, rep9b) and other mobile genetic elements (Tn917, cn_5536_ISEnfa1, Tn6009, ISEnfa1, ISEfa10) were detected. Clinically relevant E. faecium ST32 and ST416 clones were identified in meat samples. Conclusion: The occurrence of antimicrobial-resistant Enterococcus spp. in livestock and raw meat samples, carrying multiple resistance and virulence genes, including known clones associated with hospital-acquired infections, underscores the critical need for employing robust tools like whole genome sequencing. Such tools provide detailed data essential for ongoing surveillance efforts aimed at addressing the challenge of antimicrobial resistance with a focus on one health.

A review of food additives to control the proliferation and transmission of pathogenic microorganisms with emphasis on applications to raw meat-based diets for companion animals.

Raw meat-based diets (RMBDs) or sometimes described as biologically appropriate raw food (BARFs) are gaining in popularity amongst dog and cat owners. These pet guardians prefer their animals to eat minimally processed and more "natural" foods instead of highly heat-processed diets manufactured with synthetic preservatives. The market for RMBDs for dogs and cats is estimated at $33 million in the United States. This figure is likely underestimated because some pet owners feed their animals raw diets prepared at home. Despite their increasing demand, RMBDs have been plagued with numerous recalls because of contamination from foodborne pathogens like Salmonella, E. coli, or Campylobacter. Existing literature regarding mitigation strategies in RMBD's for dogs/cats are very limited. Thus, a comprehensive search for published research was conducted regarding technologies used in meat and poultry processing and raw materials tangential to this trade (e.g., meats and poultry). In this review paper, we explored multiple non-thermal processes and GRAS approved food additives that can be used as potential antimicrobials alone or in combinations to assert multiple stressors that impede microbial growth, ultimately leading to pathogen inactivation through hurdle technology. This review focuses on use of high-pressure pasteurization, organic acidulants, essential oils, and bacteriophages as possible approaches to commercially pasteurize RMBDs effectively at a relatively low cost. A summary of the different ways these technologies have been used in the past to control foodborne pathogens in meat and poultry related products and how they can be applied successfully to impede growth of enteric pathogens in commercially produced raw diets for companion animals is provided.

Zoonotic diseases risk perceptions and protective behaviors of consumers associated with consumption of meat and milk in and around Bishoftu, Ethiopia.

Consumption of raw or undercooked milk and meat is a major source of zoonoses. Information on the public risk perceptions and protective behaviors are essential for prevention and control of these diseases. However, such information is limited in Ethiopia. We assessed the risk perceptions and protective behaviors of the community toward zoonoses associated with consumption of raw meat and milk by employing the basic constructs of food safety health belief model. We collected data from 210 respondents (105 from each urban and peri-urban) using a face-to-face questionnaire interview. Majority of the respondents (96.2%) had knowledge about zoonoses. Despite their knowledge, 91% and 94.3% had raw meat and raw milk consumption habits, respectively. The odds of raw meat consumption was significantly higher in males (AOR = 3.90; CI = 1.28-11.86) and raw milk consumption was higher in females (AOR = 3.82; CI = 0.78-18.65). About 65% of respondents, obtained meat from backyard slaughtering. Self-owned and smallholder dairy farms are the major sources of milk for peri-urban and urban dwellers, respectively. While 46.7% of the respondents reported that community members are the primary sources of information, only 2.4% reported animal health professionals as their primary source of information. More than half of the respondents either moderately or strongly agreed that consumption of raw meat and raw milk can expose them to zoonoses. Urban dwellers had perceived more seriousness of consuming raw animal products. Peri-urban residents had significantly lower intention than urban ones to implement protective behaviors such as stopping consuming raw meat (P = 0.017) and milk (P = 0.043). We noted that lack of access to refrigerator and pasteurized milk were the perceived barriers for protection against zoonoses among the peri-urban dwellers. There was significant difference in perceived benefits of avoiding consumption of raw meat (P = 0.005) and milk (P = 0.001) between urban and peri-urban residents. Our study showed that irrespective of knowledge about zoonoses, consumptions of raw meat and raw milk remained common practices among the respondents. Public health education on the risk of consumption of raw milk and meat and the significance of protective behaviors using a one-health approach is critically needed to ensure meat and milk safety.

Transcriptomic and Metabolomic Profile Analysis of Muscles Reveals Pathways and Biomarkers Involved in Flavor Differences between Caged and Cage-Free Chickens.

The cage-free system has gained a lot of interest in recent years because it can offer chickens more freedom and is easier to manage compared with free-range rearing systems, but few studies have focused on the effect of the cage-free rearing system on meat quality and flavor. In this study, 44 Jianghan chickens were reared in caged or cage-free systems to explore the effect of different rearing systems on meat-eating quality. Sensory evaluation of cooked muscles showed that the leg muscle aroma, juiciness, and flavor intensity significantly improved by the cage-free rearing. The cage-free hens had significantly lower body weight, abdominal fat percentage, and meat fat content, but higher meat moisture content. The cage-free group had brighter breast muscle and redder leg muscle color 24 h after slaughter. Transcriptomic and metabolomic profile analysis of the leg muscle samples showed that the cage-free rearing changed biosynthesis pathways associated with glycogen metabolism, lipid and fatty acid biosynthesis and transport, muscle cellular type, and cellular components, which were related to raw meat quality. Different rearing systems also resulted in differences in glycolipid metabolism, lipid metabolism, and altered levels of intramuscular fat content and other flavor precursors. Pathways such as glycerolipid metabolism, adipocytokine signaling, and metabonomic pathways such as linoleic acid, glycerophospholipid, arginine, proline, and ß-alanine metabolism may be responsible for the meat quality and flavor change.

Detection and Molecular Identification of Salmonella Pathogenic Islands and Virulence Plasmid Genes of Salmonella in Xuzhou Raw Meat Products.

ABSTRACT: Virulence genes expressed in Salmonella are a primary contributing factor leading to the high morbidity and mortality of salmonellosis in humans. The pathogenicity of Salmonella is mainly determined by the specific virulence factors that it carries. These factors also confer greater virulence and play a role in infection of a host and transmission of disease, and most Salmonella enterica can cause cross-infections between humans and animals. In this study, 265 samples in total were collected from a farmer's market and two supermarkets in Xuzhou, Jiangsu province, China, including 205 pork samples and 60 chicken samples. The suspected Salmonella isolates were isolated and identified using microbiological and molecular methods, and the confirmed isolates were used for serovar analysis and antimicrobial susceptibility testing. The virulence genes of Salmonella pathogenic islands (SPIs) and Salmonella virulence plasmids (Spv) in Salmonella-positive isolates were subsequently detected. Salmonella was isolated from 29.0% of samples, and all isolates were confirmed by PCR targeting the stn gene. Among the Salmonella isolates, resistance was most frequently observed against ciprofloxacin (84.4%), followed by tetracycline (71.4%) and streptomycin (68.8%). Resistance to amoxicillin-clavulanic acid (6.3%) and aztreonam (5%) was less commonly detected. The presence of the following virulence genes was determined by specific PCRs: hilA (SPI-1), sifA (SPI-2), misL (SPI-3), siiE (SPI-4), sopB (SPI-5), and spvC. The detection rate for SPI-1 to SPI-5 was 93.5, 87.0, 97.4, 97.4, and 97.4%, respectively. In addition, the detection rate of the spvC gene was 96.1%. Except for sopB (94.7%), all isolates of the dominant serovar S. enterica subsp.. enterica serovar Enteritidis contained all virulence genes from SPI-1 to SPI-5. This study demonstrated the epidemiological status of Salmonella in raw meat products in Xuzhou, and the complex antibiotic resistance and high isolation rate of virulence genes observed reveal many potential risks of which the findings presented herein will provide orientation to improve public health safeguards.

Microbiological Quality and Presence of Foodborne Pathogens in Raw and Extruded Canine Diets and Canine Fecal Samples.

Pet food can be a source of microbiological hazards that might affect companion animals and owners. Even though owners usually rely on conventional pet diets, such as extruded diets, new feeding practices, such as raw meat-based diets (RMBDs), have grown. RMBDs' benefits are still scientifically uncertain, while its risks have been documented. The use of canine RMBDs might increase the exposure to zoonotic pathogens, such as Salmonella spp., Listeria monocytogenes, Campylobacter spp., among others. Identifying pathogen prevalence in canine food and pets is required to contribute to public health measures. The aims of this study were: (1) to compare the microbiological quality of RMBDs and extruded diets (2) to identify and compare the prevalence of Salmonella spp., Campylobacter jejuni, and L. monocytogenes from raw and extruded canine diets and canine fecal samples, and (3) to characterize pet owners according to the diet chosen to be used on their pets, their motivations for using RMBDs, and their knowledge about benefits and risks related to this feeding practice. Conventional and molecular microbiological methods were used to identify pathogen presence from food and fecal samples, while pulsed-field gel electrophoresis (PFGE) was performed to evaluate the clonal relationship between isolates. Aerobic plate counts for RMBDs were higher than those detected for extruded diets. Salmonella spp. and L. monocytogenes were isolated from 35.7% (15/42) RMBDs, while Salmonella spp., C. jejuni, and L. monocytogenes from 33.3% (11/33) fecal samples from RMBD-fed dogs. From the RMBD samples positive to Salmonella spp., chicken was the main meat ingredient composing the diets. PFGE analysis confirmed a genetic association between Salmonella spp. isolates from fecal and raw food samples from the same household. We did not detect pathogens from extruded food samples or feces from extruded-fed dogs. Using a survey, we identified dog owners' unawareness and/or underestimation of risks related to RMBDs. We demonstrated that canine raw pet food might be a source of zoonotic foodborne pathogens that represent a health risk for both humans and pets. While clinical findings caused by the mentioned pathogens vary among pets, the zoonotic potential implies a significant concern.

Isolation and identification of Helicobacter pylori from raw chicken meat in Dhamar Governorate, Yemen.

Although Helicobacter pylori (H. pylori) is one of the most common bacterial pathogens of human, its natural reservoirs are still unclear. There is an increasing number of reports that document the occurrence of H. pylori in various foods. This study aimed at isolation of H. pylori from chicken meat sampled. Two hundred and sixty samples were collected randomly from slaughterhouses and markets in Dhamar Governorate, Yemen. Samples were enriched in Brain-Heart Infusion broth in microaerophilic conditions before inoculating the Camp-Blood agar and EYE agar plates. Results showed that 13.8% of samples were contaminated evidenced by H. pylori growth via traditional culture method on agar media. No significant differences between sample types (thighs and breast muscles) (p=0.353) or the sampling source (p=0.816) were observed. Autumn season was associated with increased occurrence of H. pylori. The source of H. pylori in food is still not identified. Proper cooking and good sanitation practices are highly recommended to avoid the infection. Further studies addressing the potential sources of H. pylori are highly suggested.

From chicken to salad: Cooking salt as a potential vehicle of Salmonella spp. and Listeria monocytogenes cross-contamination.

Epidemiological studies show that improper food handling practices at home account for a significant portion of foodborne illness cases. Mishandling of raw meat during meal preparation is one of the most frequent hazardous behaviours reported in observational research studies that potentially contributes to illness occurrence, particularly through the transfer of microbial pathogens from the raw meat to ready-to-eat (RTE) foods. This study evaluated the transfer of two major foodborne pathogens, Salmonella enterica and Listeria monocytogenes, from artificially contaminated chicken meat to lettuce via cooking salt (used for seasoning) during simulated domestic handling practices. Pieces of chicken breast fillets were spiked with five different loads (from ca. 1 to 5 Log CFU/g) of a multi-strain cocktail of either S. enterica or L. monocytogenes. Hands of volunteers (gloved) contaminated by handling the chicken, stirred the cooking salt that was further used to season lettuce leaves. A total of 15 events of cross-contamination (three volunteers and five bacterial loads) were tested for each pathogen. Immediately after the events, S. enterica was isolated from all the cooking salt samples (n = 15) and from 12 samples of seasoned lettuce; whereas L. monocytogenes was isolated from 13 salt samples and from all the seasoned lettuce samples (n = 15). In addition, S. enterica and L. monocytogenes were able to survive in artificially contaminated salt (with a water activity of 0.49) for, at least, 146 days and 126 days, respectively. The ability of these foodborne pathogens to survive for a long time in cooking salt, make it a good vehicle for transmission and cross-contamination if consumers do not adopt good hygiene practices when preparing meals.

Prevalence, Antibiotic Resistance, Toxin-Typing and Genotyping of Clostridium perfringens in Raw Beef Meats Obtained from Qazvin City, Iran.

BACKGROUND: Clostridium perfringens is one of the highest prevailing spore-forming foodborne pathogens, which is widely distributed and causes severe disease and outbreaks in humans and animals. Raw meat and poultry are the main vehicles of this pathogen. In this study, we investigated the prevalence, antibiotic resistance pattern, toxin-encoding genes and genetic diversity of C. perfringens isolates from raw whole and minced meat samples purchased from local markets in Qazvin city, Iran (the source of beef cattle production was also located in Qazvin city, Iran). METHODS: We used conventional culture-based and Kirby-Bauer disk diffusion and conventional and arbitrary primer PCR methods. RESULTS: A total of 18 C. perfringens strains were isolated from 133 raw meat samples (13.53%). Up to 44.4 and 55.5% of these isolates were detected in raw minced and whole meat samples, respectively. We found that 72.2, 66.6, 61.1, 37.8 and 33.3% of the C. perfringens isolates were resistant to ampicillin, tetracycline, amoxicillin, ciprofloxacin and chloramphenicol antibiotics, respectively. Multidrug resistance was found in 38% of the isolates. Among the four main toxin genes evaluated, the Cpa gene was detected in all isolates, and 61.1% of the isolates were mostly recognized as type A C. perfringens. High levels of genetic diversity were observed among the isolates, and they were classified into five distinct groups. CONCLUSIONS: The isolates from whole meat samples were more resistant to antibiotics. However, toxin genes were more detected in the isolates from minced meat samples. Our findings suggest that contamination of raw meat products with multidrug resistant C. perfringens could be regarded as one of the concerning pathogens in these products. Comprehensive monitoring of C. perfringens isolates is strongly recommended.