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Helicobacter suis is the most common non-Helicobacter pylori gastric Helicobacter species found in humans. Infection is associated with gastritis, peptic ulcer, gastric MALT lymphoma and neurodegenerative disorders, particularly Parkinson's disease. However, the pathogenicity of this species is still a matter of research, and results of virulence studies and antibiotic susceptibility tests tend to vary between strains. Cholesterol α-glucosyltransferase (αCgT), a known H. pylori virulence factor, appears to be present in most clinical H. suis isolates. The ability to form biofilms also plays a crucial role in the pathogenesis of H. pylori. However, no reports have been published on this ability in H. suis. H. suis is considered an emerging zoonotic pathogen, with pigs being the main source of human infection. However, there is very little information on its presence in pork, mainly due to the difficulties of its culture. Therefore, our aim was to determine the prevalence of H. suis in pork products from our geographical area by PCR and Fluorescence in situ Hybridization (FISH), as well as to isolate the bacteria and determine the antibiotic susceptibility patterns, the presence of the αCgT gene and the ability of the isolates to form biofilms. Overall, H. suis was detected in 20 of the 70 (28.6 %) samples analyzed. In 3 of them, H. suis was isolated. The αCgT gene was detected in all isolates and two of them showed a multiresistance pattern. The H. suis reference strain and two of the isolates showed "strong" to "moderate" in vitro biofilm formation ability under optimal growth conditions. Our results seem to indicate that H. suis is significantly prevalent in pork products. The combination of culture with FISH and/or mPCR proved to be a rapid and specific method for the detection, identification and direct visualization of cultivable H. suis cells from pork food products., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Pathogen control in the meat industry relies on the effectiveness of postharvest interventions in reducing microbial populations. This study investigated differences in the survival of Salmonella serovars when exposed to organic acids used as antimicrobials on raw pork meat. Seven serovars were included in this study (S. Newport, S. Kentucky, S. Typhimurium, S. Dublin, S. Heidelberg, S. Infantis, and S. Enteritidis). Multistrain serovar cocktails were prepared and tested against lactic acid (LA) and peracetic acid PAA at two concentrations, LA 2 and 4% and PAA 200 and 400 ppm. Pork samples were assigned to each serovar, inoculated with 6.0 Log CFU/cm 2 Salmonella (one serovar at a time), and treated with the corresponding antimicrobials. A two-way analysis of variance was conducted to examine the effects of serovar and antimicrobial concentrations on Salmonella survival. A significant main effect of serovar was identified, indicating that Salmonella concentration and reduction rate were significantly affected by serovar. Similarly, a significant main effect of antimicrobials was observed, suggesting that the treatment types impacted Salmonella concentration and reduction rate. However, the interaction effect between serovar and antimicrobial was not significant. Posthoc comparisons indicate that PAA 400 ppm is more effective at reducing Salmonella concentrations and that S. Dublin may be more susceptible than S. Newport to antimicrobial sprays. Additionally, under PAA exposure, only S. Dublin, S. Kentucky, and S. Heidelberg showed statistically significant differences (P < 0.05) compared with the control, indicating that these three serovars are more susceptible to PAA treatments than the rest. The behavior of different Salmonella serovars under stress conditions can give us an insight into how these pathogens survive processing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonella exhibits extensive genetic diversity, facilitated by horizontal gene transfer occurring within and between species, playing a pivotal role in this diversification. Nevertheless, most studies focus on clinical and farm animal isolates, and research on the pangenome dynamics of Salmonella isolates from retail stage of the animal food supply chain is limited. Here, we investigated the genomes of 950 Salmonella isolates recovered from retail chicken and pork meats in seven provinces and one municipality of China in 2018. We observed a strong correlation between Salmonella sublineage diversity and the accessory genome with meat type, revealing reduced diversity associated with increased resistance. Importantly, genes associated with antibiotic, biocide, and heavy metal resistance were unevenly distributed in Salmonella from retail chicken and pork. Pork Salmonella isolates showed a higher prevalence of copper and silver resistance genes, while chicken Salmonella isolates displayed a significant predominance of genetic determinants associated with cephalosporin and ciprofloxacin resistance. Moreover, co-occurrence patterns of resistance determinants and their interaction with mobile genetic elements also correlated with meat type. In summary, our findings shed light on how Salmonella achieves their ecological niche success driven by evolution and gene changes in the retail stage of the animal food supply chain., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The spoilage of refrigerated pork involves nutrient depletion and the production of spoilage metabolites by spoilage bacteria, yet the microbe-metabolite interactions during this process remain unclear. This study employed 16S rRNA high-throughput sequencing and non-targeted metabolomics based on ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) to reveal the core microbiota and metabolite profiles of pork during refrigeration. A total of 45 potential biomarkers were screened through random forest model analysis. Metabolic pathway analysis indicated that eleven pathways, including biogenic amine metabolism, pentose metabolism, purine metabolism, pyrimidine metabolism, phospholipid metabolism, and fatty acid degradation, were potential mechanisms of pork spoilage. Correlation analysis revealed nine metabolites-histamine, tyramine, tryptamine, D-gluconic acid, UDP-d-glucose, xanthine, glutamine, phosphatidylcholine, and hexadecanoic acid-as spoilage biomarkers, with Pseudomonas, Serratia, and Photobacterium playing significant roles. This study provides new insights into the changes in microbial and metabolic characteristics during the spoilage of refrigerated pork., Competing Interests: Declaration of competing interest We declare that no conflict of interest exits in the submission of this manuscript, and manuscript is approved by all authors for publication., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

In order to explore the application prospects of static magnetic field (SMF) combined with supercooling in meat preservation, this study proposed a novel method of supercooling assisted by a stationary magnetic field (SMF + supercooling) for the preservation of chilled pork, evaluating its cooling rate and quality changes (e.g., water holding capacity, color, pH, and TVB-N), as well as the evolution trend of the microbiota. The results showed that SMF + supercooling significantly (P < 0.05) improved the cooling rate of pork. Compared to chilling and supercooling, SMF + supercooling significantly delayed the increase of TVB-N and TVC on the 12th day of storage (P < 0.05). SMF + supercooling treatment achieves the maintenance of pork water-holding capacity by inhibiting water migration, reducing drip loss, cooking loss, and centrifugal loss of pork. The 16S rDNA bacteria flora analysis demonstrated that SMF + supercooling treatment reduced the relative abundance of spoilage bacteria such as Acinetobacter, Streptococcus, and Pseudomonas, delaying the deterioration of pork quality caused by microbial growth. The SMF + supercooling treatment can be considered a novel refrigeration preservation method that delays the deterioration of pork quality and extends its shelf life., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Salmonella is a foodborne pathogen of global significance and is highly prevalent in pork. This study investigated the prevalence, contamination distribution, virulence genes and antibiotic resistance of Salmonella in 3 pork processors in the Shandong Province of China. Samples were collected from 13 different sampling sources across the slaughter procedures (600 samples) as well as at retail outlets supplied by these processors (45 samples). The prevalence was 18.9 % among all the samples, with the highest prevalence observed in feces (40.0 %), lairage pens (38.0 %), and hides (34.0 %). A total of 6 serotypes were identified, with S. Rissen (46.3 %) and S. typhimurium (32.0 %) found to be the most prevalent serotypes. 86.8 % of Salmonella isolates were multi-drug resistant, with the majority of strains resistant to erythromycin, sulfisoxazole, and ampicillin. The multilocus sequence typing analysis revealed 6 STs were obtained from 45 isolates, with the dominant type ST469 accounting for 40.0 % of the total, which suggested a high possibility of cross-contamination between the plant processing chain and retail outlets. This work reveals the prevalence and correlation of Salmonella isolates between pork slaughter and retail outlets and acts as a case-study for stakeholders wishing to improve pork supply chain hygiene, control cross-contamination between the various slaughtering processes, and obtain continuous updates on Salmonella surveillance., Competing Interests: Declaration of competing interest The authors declare that no conflict of interest., (Copyright © 2024. Published by Elsevier Ltd.)

Streptococcus suis, a bacterium commonly found in pigs, causes infections in humans through direct contact with infected animals or consumption of contaminated pork products. Recently, a localized outbreak of S. suis infection in humans resulted in three confirmed cases. All three patients had some form of contact with pigs in their medical history. One patient worked at a pig farm, whereas the other two consumed raw pork soup at the same restaurant. The patients were diagnosed with septicemia, subdural empyema, and infectious spondylitis. Streptococcus suis was isolated from their blood. This study was conducted to investigate the clinical features of three patients with S. suis infection and perform a molecular biological analysis of the strains obtained from them. Subsequent investigations highlighted the potential sources for this rare but serious infection and provided insight into preventive measures.

In this study, a pretreatment method based on a magnetic capture probe for the rapid isolation and enrichment of bacteria from raw pork was developed. The chitosan immobilized Fe 3 O 4 @MIL-100(Fe) was prepared as a capture probe for total bacterial counts through the electrostatic interaction of positively charged chitosan and the negatively charged substances on the surface of bacteria. The interference of matrix in pork samples on this method was studied and removed by differential centrifugation. The results showed the capture probe had a great selectivity binding and magnetic separation properties for the tested six common bacteria in pork. Under the optimal conditions, the capture efficiency of the bacteria (10 5 CFU mL -1 ) from pork surface samples was all above 90%. The capture efficiency of the bacteria in a homogenate system was greatly decreased due to the interference of sarcoplasmic protein and myofibrillar protein in pork. The matrix effect was mitigated by a differential centrifugation method, and the capture efficiency of all six bacteria was >80%. The developed magnetic separation method took 40 min and showed good isolation and enrichment properties of bacteria. Thus, the proposed method is expected to provide a simple, convenient, and time-saving pretreatment method for the detection of total bacterial counts in pork., (© 2024 Institute of Food Technologists.)

Salmonella contamination of pork products is a significant public health concern. Temperature abuse scenarios, such as inadequate refrigeration or prolonged exposure to room temperature, can enhance Salmonella proliferation. This study aimed to develop and validate models for Salmonella growth considering competition with background microbiota in raw ground pork, under isothermal and dynamic conditions of temperature abuse between 10 and 40 °C. The maximum specific growth rate (μ max ) and maximum population density (MPD) were estimated to quantitatively describe the growth behavior of Salmonella. To reflect more realistic microbial interactions in Salmonella-contaminated product, our model considered competition with the background microbiota, measured as mesophilic aerobic plate counts (APC). Notably, the μ max of Salmonella in low-fat samples (∼5 %) was significantly higher (p < 0.05) than that in high-fat samples (∼25 %) at 10, 20, and 30 °C. The average doubling time of Salmonella was 26, 4, 2, 1.5, 0.8, and 1.1 h at 10, 15, 20, 25, 30, and 40 °C, respectively. The initial concentration of Salmonella minimally impacted its growth in ground pork at any temperature. The MPD of APC consistently exceeded that of Salmonella, indicating the growth of APC without competition from Salmonella. The competition model exhibited excellent fit with the experimental data, as 95 % (627/660) of residual errors fell within the desired acceptable prediction zone (pAPZ >0.70). The theoretical minimum and optimum growth temperatures for Salmonella ranged from 5 to 6 °C and 35 to 36 °C, respectively. The dynamic model displayed strong predictive performance, with 90 % (57/63) of residual errors falling within the APZ. Dynamic models could be valuable tools for validating and refining simpler static or isothermal models, ultimately improving their predictive capabilities to enhance food safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The degree of contamination of retail meat with colistin-resistant bacteria and its potential contribution to dissemination within communities remains to be determined. Thus, we aimed to elucidate the contamination status of colistin-resistance genes, indicative of colistin-resistant bacteria, in retail meats in Vietnam. In total, 46 chicken and 49 pork meats from stores in Vietnam and Japan were examined. Multiplex real-time polymerase chain reaction with TaqMan probes was performed for detecting mcr-1 , mcr-3 , and Escherichia coli 16S rRNA . Colistin-resistant bacteria in meats were isolated using selective media. The minimum inhibitory concentrations of colistin were determined using the broth microdilution method. The results showed that 70.7% of chicken meats in Vietnam were contaminated with both mcr-1 and mcr-3 . Meanwhile, mcr-1 and mcr-3 were detected in 15.9% and 40.9% of pork meat, respectively. Only mcr-3 was detected in 40% of chicken in Japan. In addition, mcr-1 -harboring E. coli and mcr-3 -harboring Aeromonas were isolated from chicken meats in Vietnam. Some of these isolates showed colistin resistance. These results showed that most retail meats were highly contaminated with colistin-resistance genes. Notably, our results suggest that mcr-3 is more prevalent in the contaminated samples compared with mcr-1 .

This study compared the shelf-life of beef and pork longissimus lumborum muscles (loins) that had the same initial bacterial loads and were held under the same chilled storage conditions. To identify the underlying pathways, comparisons were conducted from the perspective of the spoilage indicators; protease/lipase activity, and the volatile organic compounds (VOC) generated over 28 d of chilled storage. The initial total viable microbial count (TVC) on Day 0 for both type of meat was 4.3 log 10 CFU/g. It was found that the TVC of beef and pork did not differ throughout the total chilled storage period and both ultimately exceeded 7 log 10 CFU/g after 28 d. Based on total volatile basic nitrogen (TVB-N) guidelines, pork was spoilt after 21 d of chilled storage and therefore 7 d earlier than beef. Changes in the concentration of VOC spoilage biomarkers, including 1-octen-3-ol, 1-octanol, nonanal, and others, confirmed that pork had a shorter shelf-life than beef. An important reason for the difference in shelf-life between the two types of meat was that pork had a higher protease activity, although the beef had higher levels of total lipase activity. These findings help us understand the differences in the spoilage process of raw meat from different species and explore specific measures to control the spoilage of beef or pork., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024. Published by Elsevier Ltd.)

Volatile organic compounds (VOCs) indicative of pork microbial spoilage can be quantified rapidly at trace levels using selected-ion flow-tube mass spectrometry (SIFT-MS). Packaging atmosphere is one of the factors influencing VOC production patterns during storage. On this basis, machine learning would help to process complex volatolomic data and predict pork microbial quality efficiently. This study focused on (1) investigating model generalizability based on different nested cross-validation settings, and (2) comparing the predictive power and feature importance of nine algorithms, including Artificial Neural Network (ANN), k-Nearest Neighbors, Support Vector Regression, Decision Tree, Partial Least Squares Regression, and four ensemble learning models. The datasets used contain 37 VOCs' concentrations (input) and total plate counts (TPC, output) of 350 pork samples with different storage times, including 225 pork loin samples stored under three high-O 2 and three low-O 2 conditions, and 125 commercially packaged products. An appropriate choice of cross-validation strategies resulted in trustworthy and relevant predictions. When trained on all possible selections of two high-O 2 and two low-O 2 conditions, ANNs produced satisfactory TPC predictions of unseen test scenarios (one high-O 2 condition, one low-O 2 condition, and the commercial products). ANN-based bagging outperformed other employed models, when TPC exceeded ca. 6 log CFU/g. VOCs including benzaldehyde, 3-methyl-1-butanol, ethanol and methyl mercaptan were identified with high feature importance. This elaborated case study illustrates great prospects of real-time detection techniques and machine learning in meat quality prediction. Further investigations on handling low VOC levels would enhance the model performance and decision making in commercial meat quality control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The connection and temperature control of cold chain links are key to ensuring meat quality. Considering the practical production of cold chain logistics in China, this study investigated the impact of various cold chain logistics modes (including warehousing, transportation, and sales conditions) at different temperature settings (7 °C, 4 °C, and - 1 °C), modeled in the laboratory, on the quality and bacterial community succession of pork. The pork quality was evaluated by pH, water holding capacity, total volatile basic nitrogen (TVB-N), total viable count (TVC) and myowater status. Among the different cold chain logistics modes, the LL1 (samples being warehoused and transported at 4 °C for 96 h and sold at -1 °C) and the SL1 (samples being warehoused and transported at 4 °C for 30 h and sold at -1 °C) modes were suitable for inter-provincial and intra-provincial transportation due to their long shelf life (> 14 days), respectively. The bacterial community succession of pork in different cold chain logistics modes was accessed by high-throughput sequencing. The results indicated that the cold chain logistics modes had affected the bacterial community, with Latilactobacillus being the dominant bacteria in the LL1 mode and SL1 mode during spoilage. The study revealed that the entire or partial process supercooling treatment (-1 °C) during the cold chain logistics process could effectively preserve the meat quality, supporting the high-quality development of the fresh meat cold chain logistics., Competing Interests: Declaration of competing interest The authors declared that they have no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Shigella flexneri has the ability to contaminate pork and cause foodborne diseases. This study aimed to examine the effectiveness of linalool (a natural preservative) against S. flexneri and explore its potential application in contaminated pork. The results showed that linalool was capable of damaging the cell membrane and binding to the DNA of S. flexneri, and inhibiting biofilm formation and disrupting mature biofilms. The antibacterial effectiveness of linalool on the surface of pork was further demonstrated by analyzing the physicochemical properties of the pork (i.e., weight loss rate, pH value, color index, and TVB-N value) and its protein profiles. Linalool did not completely kill S. flexneri in pork at minimum bactericidal concentration (MBC) concentration and its antibacterial effect of linalool was stronger during the initial stage of storage. During storage, linalool influenced the abundance of specific proteins in the pork, particularly those involved in pathways related to fat metabolism. These findings offer novel insights into the antibacterial efficacy of linalool and its underlying mechanism in pork., Competing Interests: Declaration of competing interest The authors declares no conflict of interest for this research work., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The pork production chain is an important reservoir of antimicrobial resistant bacteria. This study identified and characterized integrons in Salmonella isolates from a Brazilian pork production chain and associate them with their antibiotic resistance pattern. A total of 41 whole-genome sequencing data of nontyphoidal Salmonella were analyzed using PlasmidSPAdes and IntegronFinder software. Nine isolates (21.9%) had some integrons identified (complete and/or incomplete). Six complete class 1 integrons were found, with streptomycin resistance genes ( aadA1 , aadA2 ) alone or downstream of a trimethoprim resistance gene ( dfrA1 , dfrA12) , and some also containing resistance genes for sulfonamides ( sul1 , sul3 ) and chloramphenicol ( cmlA1 ). Class 2 integron was detected in only one isolate, containing dfrA1 - sat2 - aadA1 gene cassettes. Five isolates harbored CALINs-clusters attC but lacking integrases-with antimicrobial resistance genes typically found in integron structures. In all, integrons were observed among four serotypes: Derby, Bredeney, Panama, and monophasic var. Typhimurium I 4,[5],12:i:-. The association of integrons with antibiotic resistance phenotype showed that these elements were predominantly identified in multidrug resistance isolates, and six of the seven gentamicin-resistant isolates had integrons. So, surveillance of integrons in Salmonella should be performed to identify the potential for the spread of antimicrobial resistance genes among bacteria.

Introduction: An outbreak of gastroenteritis due to Salmonella Give, a very rarely identified serotype in human isolates in Greece, occurred in participants of a religious festival in a rural area of southern Greece, in September 2022. The objectives of this study were to describe the outbreak in terms of epidemiology, identify the vehicle of transmission of the foodborne pathogen and recommend prevention measures., Methods: The outbreak was linked to the consumption of a local traditional recipe of roasted pork meat served by a street food vendor. In 2018, the same food item, served in a restaurant in the same region, was implicated in another S. Give outbreak., Results: Outbreak investigations revealed that outbreak-associated isolates, of food and human origin, belonged to the same S. Give strain. Significant deficiencies regarding food safety practices were identified., Conclusion: Technical knowledge about pathogen transmission paths is important in order for both food handlers and consumers to follow hygiene and sanitary measures, mainly in cases of mass gatherings, where large quantities of food are prepared, handled, cooked and served. Efficient official supervision, mainly during summer festivals, is required in order to avoid recurrence of foodborne infections by different combinations of pathogens/food commodities.

To clarify the relationship between microorganisms and physicochemical indicators of Xuanwei ham. Six ham samples for the first, second and third year were selected, respectively. The changes of physicochemical properties, the free fatty acids and microbial communities of Xuanwei ham were investigated by GC-MS and high-throughput sequencing technology. Results showed that scores of colour, overall acceptability, texture, taste and aroma were the highest in the third year sample. With increasing ripening time, moisture content, water activity (Aw), lightness (L*), springiness, and resilience decreased continuously, and yellowness (b*) was the highest in the second year sample. 31 free fatty acids were detected, and unsaturated fatty acids such as palmitoleic acid, oleic acid, and linoleic acid were the major fatty acids. The content of palmitoleic acid, oleic acid and eicosenoic acid increased significantly during processing. At the phylum level, the dominant bacteria were Proteobacteria and Firmicutes, and fungi were Ascomycota. At the genus level, the dominant bacteria were Staphylococcus and Psychrobacter, and fungi were Aspergillus. Correlation analysis showed that water content and Aw were closely related to microorganisms, and most unsaturated fatty acids were significantly correlated with microorganisms. These findings showed that microorganisms played an important role in the quality of Xuanwei ham, and provided a scientific basis for the quality control of Xuanwei ham., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Here, we developed a nano-TiO 2 -nisin-modified chitosan composite packaging film and investigated its properties and antibacterial activity, as well as its effect on chilled pork preservation time. The results indicated that the preservation time of chilled pork coated with a nano-TiO 2 -nisin-modified chitosan film (including 0.7 g/L nano-TiO 2 , irradiated with ultraviolet light for 40 min, and dried for 6 h) followed by modified atmosphere packaging (50% CO 2  + 50% N 2 ) increased from 7 to 20 days at 4 °C. Both nano-TiO 2 and nisin enhanced the mechanical strength of the chitosan film, and nisin promoted nano-TiO 2 dispersion and compatibility in chitosan. Treatment with 0.4 g/L nano-TiO 2 for 60 min considerably inhibited spoilage bacteria, particularly Acinetobacter johnnii XBB1 (A. johnnii XBB1). As nano-TiO 2 concentration and photocatalytic time increased, K + , Ca 2+ , and Mg 2+ leakage in A. johnnii XBB1 increased but Na + /K + -ATPase and Ca 2+ /Mg 2+ -ATPase activities decreased. In A. johnnii XBB1, TiO 2 significantly downregulated the expression of putrefaction-related genes such as cysM and inhibited cell self-regulation and membrane wall system repair. Therefore, our nano-TiO 2 -nisin-modified chitosan film could extend the shelf life without the addition of any chemical preservatives, demonstrating great potential for application in food preservation., Competing Interests: Declaration of competing interest The authors report no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This study aimed to investigate the potential mechanisms associated with the persistence of chloramphenicol (CHP) resistance in Escherichia coli and Salmonella enterica isolated from pigs, pork, and humans in Thailand. The CHP-resistant E. coli (n = 106) and Salmonella (n = 57) isolates were tested for their CHP susceptibility in the presence and absence of phenylalanine arginine β-naphthylamide (PAβN). The potential co-selection of CHP resistance was investigated through conjugation experiments. Whole genome sequencing (WGS) was performed to analyze the E. coli (E329, E333, and E290) and Salmonella (SA448, SA461, and SA515) isolates with high CHP MIC (32-256 μg/mL) and predominant plasmid replicon types. The presence of PAβN significantly reduced the CHP MICs (≥4-fold) in most E. coli (67.9%) and Salmonella (64.9%). Ampicillin, tetracycline, and streptomycin co-selected for CHP-resistant Salmonella and E. coli-transconjugants carrying cmlA. IncF plasmids were mostly detected in cmlA carrying Salmonella (IncFIIAs) and E. coli (IncFIB and IncF) transconjugants. The WGS analysis revealed that class1 integrons with cmlA1 gene cassette flanked by IS26 and TnAs1 were located on IncX1 plasmid, IncFIA(HI1)/HI1B plasmids and IncFII/FIB plasmids. IncFIA(HI1)/HI1B/Q1in SA448 contained catA flanked by IS1B and TnAs3. In conclusion, cross resistance through proton motive force-dependent mechanisms and co-selection by other antimicrobial agents involved the persistence of CHP-resistance in E. coli in this collection. Dissemination of CHP-resistance genes was potentially facilitated by mobilization via mobile genetic elements., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Puangseree et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Nontyphoidal Salmonella is one of the major causes of self-limiting diarrheal disease and the most common foodborne pathogen worldwide. It is an important contributor to the burden of foodborne illness in South America, including Peru, where chicken and pork are important vehicles for Salmonella infection. Salmonella infections are underreported, particularly in low- and middle-income countries where concerted action tackling Salmonella along the chicken and pork chains, from primary production to retail, is urgently needed. To support and inform the implementation of new strategies to reduce Salmonella contamination of chicken and pork, this study describes the frequency and distribution of foodborne outbreaks attributed to Salmonella in Peru and evaluates the level of Salmonella in chicken and pork meat sold in markets of three regions of Peru. To that end, we analyzed historical reports of foodborne outbreaks, levels of Salmonella in chicken and pork sold in markets, and the number of mesophiles in the collected meat samples. As a result, the microbiological analysis reveals a widespread contamination of chicken (77.1%) and pork (26.8%) with Salmonella. It also pinpoints Salmonella as the causative agent in nearly half of the outbreaks (47.0%) where the potential origin is identified over a 11-year period with chicken, mayonnaise, and pork being the most likely food vehicles. These results suggest that Salmonella is a major contributor to foodborne illness in Peru and that the monitoring of mesophiles could be a good strategy for surveillance, generating data to support source attribution studies and ultimately evidence-informed policies.

Dry-cured meat products are gaining attention owing to their distinctive sensory characteristics and health benefits. In this study, two Debaryomyces hansenii strains were investigated for their potential as starter cultures for dry-cured pork belly products. After preliminary screening, these D. hansenii strains, namely, S20 and S26, both exhibiting with excellent aroma-producing capacity in a dry-cured meat model, were selected as single-strain starter cultures. For comparison, a non-inoculated control was also evaluated. In S20- and S26-inoculated pork belly, yeast dominated the microbiota and improved microbiological safety by suppressing Enterobacteriaceae growth. Compared with the non-inoculated control, the inoculated pork belly yielded higher hardness and redness (a*) values. Starter culture inoculation accelerated proteolysis in pork belly, improving the content of total free amino acids (TFFAs) and several essential free amino acids (Thr, Val, Met, Ile, Leu, and Phe) at the end of processing. Moreover, the inoculated samples exhibited higher levels of fat oxidation-derived aldehydes as well as esters, acids, alcohols and other compounds than the non-inoculated control at the end of the 95-day ripening period. Overall, these findings provide new insights into the application of D. hansenii isolated from dry-cured ham to dry-cured pork belly., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Salmonella enterica causes severe food-borne infections through contamination of the food supply chain. Its evolution has been associated with human activities, especially animal husbandry. Advances in intensive farming and global transportation have substantially reshaped the pig industry, but their impact on the evolution of associated zoonotic pathogens such as S. enterica remains unresolved. Here we investigated the population fluctuation, accumulation of antimicrobial resistance genes and international serovar Choleraesuis transmission of nine pig-enriched S. enterica populations comprising more than 9,000 genomes. Most changes were found to be attributable to the developments of the modern pig industry. All pig-enriched salmonellae experienced host transfers in pigs and/or population expansions over the past century, with pigs and pork having become the main sources of S. enterica transmissions to other hosts. Overall, our analysis revealed strong associations between the transmission of pig-enriched salmonellae and the global pork trade., (© 2024. The Author(s).)

Helicobacter pylori, linked to gastric diseases, is targeted for probiotic treatment through bacteriocin production. Bacteriocins have gained recognition for their non-toxic effects on host cells and their ability to combat a wide range of pathogens. This study aimed to taxonomically characterize and evaluate the safety and probiotic properties of the novel species of Lactococcus sp. NH2-7C isolated from fermented pork, as well as its bacteriocin NH2-7C, both in vitro and in silico. Comparative genotypic analysis revealed an average nucleotide identity of 94.96%, an average amino acid identity of 94.29%, and a digital DNA-DNA hybridization value of 63.80% when compared to Lactococcus lactis subsp. lactis JCM 5805 T . These findings suggest that strain NH2-7C represents a novel species within the genus Lactococcus. In silico assessments confirmed the non-pathogenic nature of strain NH2-7C and the absence of genes associated with virulence and biogenic amine formation. Whole-genome analysis revealed the presence of the nisA gene responsible for nisin A production, indicating its potential as a beneficial compound with anti-Helicobacter pylori activity and non-toxic characteristics. Probiotic assessments indicated bile salt hydrolase and cholesterol assimilation activities, along with the modulation of interleukin-6 and tumour necrosis factor-α secretion. Strain NH2-7C demonstrated gastrointestinal tolerance and the ability to adhere to Caco-2 cells, affirming its safety and probiotic potential. Additionally, its ability to produce bacteriocins supports its suitability as a functional probiotic strain with therapeutic potential. However, further in vitro and in vivo investigations are crucial to ensure its safety and explore potential applications for Lactococcus sp. NH2-7C as a probiotic agent., (© 2023. The Author(s).)

Background: Non-typhoidal Salmonella (NTS) is a serious foodborne pathogen that has previously been isolated from pigs presented for slaughter in a rural pork value chain in western Kenya., Methods: To understand varying NTS contamination along the value chain we assessed prevalence at slaughter, transport and retail. Suspect isolates from culture were confirmed using matrix-assisted laser desorption-ionization time of flight mass spectrometry., Results: Prevalence on pig carcasses, meat transportation containers, retailed raw and cooked pork and accompanying side salads was 18.1%, 23.9%, 28.0%, 1.9% and 8.6%, respectively., Conclusion: NTS contamination is propagated along the pork value chain in rural western Kenya, demonstrating the need for improved hygiene measures to prevent human exposure., (© The Author(s) 2023. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.)

A host-guest colorimetric strategy is described for the detection of Listeria monocytogenes (L. monocytogenes). The optical probes were self-assembled based on the supramolecular interactions between the carbonyl groups of cucurbit[7]uril portals and gold nanoparticles (CB[7]-AuNPs). Aptamer and urease modified magnetic nanoparticles were used to specifically recognize and binding to L. monocytogenes, simultaneously hydrolyzing urea to produce ammonium ion (NH 4 + ) that can reverse CB[7] induced AuNPs aggregation. In the presence of L. monocytogenes, the above-mentioned magnetic conjugates preferentially bind to the bacterial surface, which results in blocking the catalytic active sites, thus inhibiting the production of ammonium ions. The normalized absorbance ratio of A 700 nm /A 525 nm was proportional to the L. monocytogenes concentration ranging from 10 to 10 6  cfu·mL -1 , and the visual determination can be done down to 10 cfu·mL -1 . For spiked food samples analyzed without pre-enrichment, recoveries of 98.4% to 99.3% were achieved could be verified and RSD were less than 10%. This work may offer a broad prospect for sensitive and specific determination  of pathogens., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Austria, part of Springer Nature.)

Mulberry (Morus alba L.), and above all the extract from the leaves of this plant, is a natural medicine that has been used in traditional medicine for hundreds of years. Mulberry leaves contains polyphenol compounds: flavonoids, coumarins, numerous phenolic acids, as well as terpenes and steroids. The antioxidant effect of these compounds may be beneficial to the fat fraction of meat products, thereby increasing their functional qualities. The aim of the study was to evaluate the effectiveness of the use of mulberry water leaf extract, as an additive limiting adverse fat changes and affecting the functionality in model liver pâtés. Pork pâtés were prepared by replacing 20% of animal fat with rapeseed oil (RO), and water extract of mulberry leaves was added in the proportion of 0.2%, 0.6% and 1.0%. It has been shown that the addition of mulberry leaf extract delayed the appearance of primary and secondary fat oxidation products. The most effective antioxidant effect during 15-day storage was observed in the sample with the addition of 0.6% and 1.0% water mulberry leaf extract. These samples also showed inhibiting activity against angiotensin-converting enzymes and cholinesterase's. During storage, the tested pâtés had a high sensory quality with unchanged microbiological quality. Mulberry leaf extract can be an interesting addition to the production of fat meat products, delaying adverse changes in the lipid fraction and increasing the functionality of products., Competing Interests: The authors have declared that no competing interests exist.

In this study, we aimed to characterize the distribution of Yersinia enterocolitica in a pork production chain in Brazil, as well as the virulence profile and antibiotic resistance of the obtained isolates. Samples from 10 pig lots obtained from finishing farms (water, feed, and barn floors, n = 30), slaughterhouse (lairage floors, carcasses at four processing steps, tonsils, and mesenteric lymph nodes, n = 610), and processing (end cuts, processing environment, n = 160) were obtained in Paraná state, Brazil, and subjected to Y. enterocolitica detection by ISO 10,273. The obtained isolates were identified based on biochemical and molecular features (16 s rRNA, inv, bioserotyping) and subjected to PCR assays to detect virulence (ail, ystA, ystB, virF, myfA, fepA, fepD, fes, tccC, ymoA, hreP, and sat) and multidrug resistance-related genes (emrD, yfhD, and marC). Also, isolates were subjected to disk diffusion test to characterize their resistance against 17 antibiotics from 11 classes and to pulsed field gel electrophoresis (PFGE) after XbaI macro-restriction. Y. enterocolitica was detected in a single sample (tonsil), and the obtained three isolates were characterized as serotype O:3, harboring ail, ystA, virF, myfA, tccC, ymoA, hreP, emrD, yfhD, and marC, and resistant to all tested antibiotics. The three isolates presented identical macro-restriction profiles by PFGE, also identical to isolates obtained from Minas Gerais, other Brazilian state; one selected isolate was identified as biotype 4. Despite the low occurrence of Y. enterocolitica in the studied pork production, the virulence potential and the antibiotic resistance profiles of the isolates demonstrated their pathogenic potential, and the macro-restriction profiles indicate strains descending from a common subtype in the pork production chain of two Brazilian States., (© 2021. Sociedade Brasileira de Microbiologia.)

Nine odorless laboratory-collected hydro-distilled aqueous extracts (basil, calendula, centrifuged oregano, corn silk, laurel, oregano, rosemary, spearmint, thyme) and one industrial steam-distilled oregano hydrolate acquired as by-products of essential oils purification were screened for their in vitro antimicrobial activity against three Salmonella Typhimurium strains (4/74, FS8, FS115) at 4 and 37 °C. Susceptibility to the extracts was mainly plant- and temperature-dependent, though strain dependent effects were also observed. Industrial oregano hydrolate eliminated strains immediately after inoculation, exhibiting the highest antimicrobial potential. Hydro-distilled extracts eliminated/reduced Salmonella levels during incubation at 4 °C. At 37 °C, oregano, centrifuged oregano, thyme, calendula and basil were bactericidal while spearmint, rosemary and corn silk bacteriostatic. A strain-dependent effect was observed for laurel. The individual or combined effect of marinades and edible coatings prepared of industrial hydrolate and hydro-distilled oregano extracts with or without oregano essential oil (OEO) was tested in pork meat at 4 °C inoculated with FS8 strain. Lower in situ activity was observed compared to in vitro assays. Marinades and edible coatings prepared of industrial oregano hydrolate + OEO were the most efficient in inhibiting pathogen. Marination in oregano extract and subsequent coating with either 50% oregano extract + OEO or water + OEO enhanced the performance of oregano extract. In conclusion, by-products of oregano essential oil purification may be promising alternative antimicrobials to pork meat stored under refrigeration when applied in the context of multiple hurdle approach., (© 2021. The Author(s).)

Plant-derived antimicrobial agents have adequate antimicrobial effects on food-borne pathogens, which can be used as food preservatives. The purpose of this study was to evaluate the antibacterial mechanism of chlorogenic acid (CA) against Yersinia enterocolitica and Enterobacter sakazakii . The minimum inhibitory concentration (MIC) of CA was determined by employing the broth microdilution method. Then, the cell function and morphological changes of Y. enterocolitica and E. sakazakii treated with CA were characterized. Finally, the growth inhibition models of Y . enterocolitica in raw pork and E. sakazakii in skim milk were constructed through the response surface methodology. The results demonstrated that CA has a satisfactory inhibitory effect against Y. enterocolitica and E. sakazakii with a MIC of 2.5 mg/mL. In addition, CA inhibited the growth of Y. enterocolitica and E. sakazakii via cell membrane damage, such as depolarization of the cell membrane, reduction in intracellular adenosine triphosphate (ATP) and pH levels, and destruction of cell morphology. Moreover, CA reduced two log cycles of Y. enterocolitica in raw pork and E. sakazakii in skim milk at a certain temperature. According to the corresponding findings, CA has the potential to be developed as an effective preservative to control Y. enterocolitica and E. sakazakii -associated foodborne diseases.

Abstract: Shiga toxin-producing Escherichia coli (STEC) O157:H7/nonmotile and some non-O157 STEC strains are foodborne pathogens. In response to pork-associated O157 STEC outbreaks in Canada, we investigated the occurrence of STEC in Canadian retail raw ground pork during the period of 1 November 2014 to 31 March 2016. Isolated STEC strains were characterized to determine the Shiga toxin gene (stx) subtype and the presence of virulence genes encoding intimin (eae) and enterohemorrhagic E. coli hemolysin (hlyA). O157 STEC and non-O157 STEC strains were isolated from 1 (0.11%) of 879 and 13 (2.24%) of 580 pork samples, respectively. STEC virulence gene profiles containing both eae and hlyA were found only in the O157 STEC (stx2a, eae, hlyA) isolate. The eae gene was absent from all non-O157 STEC isolates. Of the 13 non-O157 STEC isolates, two virulence genes of stx1a and hlyA were found in four (30.8%) O91:H14 STEC isolates, whereas one virulence gene of stx2e, stx1a, and stx2a was identified in five (38.5%), two (15.4%), and one (7.7%) STEC isolates, respectively, of various serotypes. The remaining non-O157 STEC isolate carried stx2, but the subtype is unknown because this isolate could not be recovered for sequencing. O91:H14 STEC (stx1a, hlyA) was previously reported in association with diarrheal illnesses, whereas the other non-O157 STEC isolates identified in this study are not known to be associated with severe human illnesses. Virulence gene profiles identified in this study indicate that the occurrence of non-O157 STEC capable of causing severe human illness is rare in Canadian retail pork. However, O157 STEC in ground pork can occasionally occur; therefore, education regarding the potential risks associated with STEC contamination of pork would be beneficial for the public and those in the food industry to help reduce foodborne illnesses., (Published 2021 by the International Association for Food Protection. Copyright ©, Her Majesty the Queen, in Right of Canada as represented by the Canadian Food Inspection Agency.)

Listeria monocytogenes is an important foodborne pathogen, which imposes great burdens on public health. The current methods for detecting L. monocytogene are limited in several ways such as time consuming and lab equipment dependent. In this study, we developed a new electrochemical assay to improve the efficacy. This assay allows us to generate numerous G-quadruplex sequences while loop-mediated isothermal amplification happens. Then, these G-quadruplex sequences form DNAzyme to produce a color change and an electrochemical signal by oxidizing tetramethylbenzidine. This assay could be finished in 2 h, which significantly reduced the detection time. Also, we confirmed the limit of detection of this assay at 6.8 CFU/mL according to 3σ criterion. Our assay shows good sensitivity to detect bacteria range from 52.5 to 5.25 × 10 4 CFU/mL. This assay's reliability was also confirmed by detecting artificially contaminated pork samples. Thus, we propose this electrochemical assay for rapid and sensitive detection of L. monocytogenes in food., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Genomic diversity of Listeria monocytogenes isolates from the deboning and slicing areas of three dry-cured ham processing plants was analysed. L. monocytogenes was detected in 58 out of 491 samples from the environment and equipment surfaces, all from the deboning area, with differences in prevalence among facilities. The most frequent PCR-serogroup was IIa (74.1%) followed by IIb and IIc, and only one isolate was serogroup IVb. Twenty different pulsotypes and 11 sequence types (STs) grouped into 10 clonal complexes (CCs) were determined. ST121 (CC121) and ST9 (CC9) were the most abundant. Premature stop codons (PMSC6 and PMSC19) associated with attenuated virulence were found in the inlA sequence in 7 out of 12 selected strains. CC121 strains were strong biofilm formers and some harboured the transposon Tn6188, related with increased tolerance to quaternary ammonium compounds. L. monocytogenes clones considered hypovirulent resulted predominant in the deboning areas. The clonal structure and potential virulence of the isolates could help to establish adequate control measures and cleaning protocols for the comprehensive elimination of the pathogen in dry-cured ham processing environment., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Various methods have been described to isolate third generation cephalosporin (3GC) resistant Enterobacteriaceae from foods, but it is not known how comparable they are between studies. Here, the performance of five enrichment broths and two selective agars are compared for their ability to isolate 3GC resistant Enterobacteriaceae from retail chicken, beef, pork, and veal samples. The results showed equivalence between Enterobacteriaceae enrichment broth (EE), lauryl sulfate broth (LST), and modified typtone soy broth (mTSB). Lower isolation rates were observed when LST and mTSB were supplemented with the 3GC antibiotic cefotaxime. The overall performance of MacConkey agar supplemented with cefotaxime and a proprietary selective agar (ESBL CHROMagar) was equivalent, although differences linked to the microbiota of specific meat commodities were noted. Regardless of the isolation method, further screening was required to confirm the taxonomy and resistance of the presumptive positive strains. Approximately 40% of confirmed 3GC resistant foodborne Enterobacteriaceae strains tested positive for extended spectrum beta-lactamase (ESBL) activity. Strains that were resistant to ceftriaxone and susceptible to cefoxitin were more likely to test positive for ESBL activity, as were strains that possessed either of two ESBL genes (bla SHV or bla TEM ). Based on our results, we recommend using an antibiotic-free enrichment broth, two selective agars, and an isolate screening strategy to isolate 3GC resistant Enterobacteriaceae from retail meats. Antibiotic susceptibility testing and/or PCR screening for bla SHV or bla TEM can then be used to identify ESBL producing strains among the 3GC resistant meat isolates. The adoption of this approach by the research community will enable more effective monitoring of antibiotic resistance rates and trends among foodborne Enterobacteriaceae over time and across jurisdictions., (Crown Copyright © 2021. Published by Elsevier B.V. All rights reserved.)

The ripening process of dry-cured meat products is characterised by the development of fungi on the product's surface. This population plays a beneficial role, but, uncontrolled moulds represent a health risk, since some of them may produce mycotoxins, such as ochratoxin A (OTA). The aim of the present work is to assess the potential of near-infrared spectroscopy (NIRS) for the detection of OTA-producing mould species on dry-cured ham-based agar. The collected spectra were used to develop Support Vector Machines-Discriminant Analysis (SVM-DA) models by a hierarchical approach. Firstly, an SVM-DA model was tested to discriminate OTA and non-OTA producers; then, two models were tested to discriminate species among the OTA producers and the non-OTA producers. OTA and non-OTA-producing moulds were discriminated with 85% sensitivity and 86% specificity in the prediction. Furthermore, the SVM-DA model could differentiate non-OTA-producing species with a 95% sensitivity and specificity. Promising results were obtained for the prediction of the four OTA-producing species tested, with a 69% and 90% sensitivity and specificity, respectively. The preliminary approach demonstrated the high potential of NIR spectroscopy, coupled with Chemometrics, to be used as a real-time automated routine monitorization of dry-cured ham surfaces.

In this work, two combinations of double EOs, i.e., α-terpineol: eugenol (α-T:Eu) and carvacrol:eugenol (CA:Eu), are used to develop the active antibacterial films of double EOs@yam starch/microcrystalline cellulose (EOs@SC). The hydrogen-bonded networks in SC matrix are conducive to thermostability enhancement and the film of SC25 is determined for EO incorporation. The interactions between EOs and SC matrix are also hydrogen bonds and the double EOs@SC are smooth at ratio of ≤2:2 for α-T:Eu or CA:Eu. The ultimate film properties are dependent on the incorporated EOs. The release of EOs is well controlled by two mechanisms of diffusion (predominant) and swelling (secondary). Synergetic antibacterial activity occurs on double EOs@SC. The shelf life of pork can be extended by 1 day at 25 °C by the two typical films of α-T 2 :Eu 2 @SC and CA 2 :Eu 2 @SC. Moreover, EOs@SC can be well degraded in humus soil. Thereby, the target films will have great potential in active packaging to extend the shelf life of food., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Colistin and carbapenem-resistant bacteria have emerged and become a serious public health concern, but their epidemiological data is still limited., Objectives: This study examined colistin and carbapenem resistance in Escherichia coli and Salmonella from pigs, pig carcasses, and pork in Thailand, Lao PDR, and Cambodia border provinces., Methods: The phenotypic and genotypic resistance to colistin and meropenem was determined in E. coli and Salmonella obtained from pigs, pig carcasses, and pork (n = 1,619). A conjugative experiment was performed in all isolates carrying the mcr gene (s) (n = 68). The plasmid replicon type was determined in the isolates carrying a conjugative plasmid with mcr by PCR-based replicon typing (n = 7). The genetic relatedness of mcr -positive Salmonella (n = 11) was investigated by multi-locus sequence typing., Results: Colistin resistance was more common in E. coli (8%) than Salmonella (1%). The highest resistance rate was found in E. coli (17.8%) and Salmonella (1.7%) from Cambodia. Colistin-resistance genes, mcr-1 , mcr-3 , and mcr-5 , were identified, of which mcr-1 and mcr-3 were predominant in E. coli (5.8%) and Salmonella (1.7%), respectively. The mcr-5 gene was observed in E. coli from pork in Cambodia. Two colistin-susceptible pig isolates from Thailand carried both mcr-1 and mcr-3 . Seven E. coli and Salmonella isolates contained mcr-1 or mcr-3 associated with the IncF and IncI plasmids. The mcr -positive Salmonella from Thailand and Cambodia were categorized into two clusters with 94%-97% similarity. None of these clusters was meropenem resistant., Conclusions: Colistin-resistant E. coli and Salmonella were distributed in pigs, pig carcasses, and pork in the border areas. Undivided-One Health collaboration is needed to address the issue., Competing Interests: The authors declare no conflicts of interest., (© 2021 The Korean Society of Veterinary Science.)

To identify the microbial community and origin of the spoilage flora of bacon, the changes in microbial population numbers and community structure were followed along the processing line, using culture-independent and culture-dependent methods. 16S rRNA gene amplicon sequencing (16S-seq) analysis showed that community complexity and structure significantly differed at different processing stages. Some 428 bacterial groups were ascertained at genus level, and Acinetobacter, Pseudomonas, Psychrobacter, and Brochothrix were the predominant bacteria on raw meats. After curing specimens dominated by Psychrobacter, Weissella, Vibrio, Leuconostoc, Myroides, Acinetobacter, and Lactobacillus, a total of 33 species were identified by traditional microbiological analyses and direct sequence determination methods. Our results indicated that curing should be considered one of the primary factors during various processing steps, presumably contaminating the products directly or indirectly., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Species belonging to the genus Sphingomonas have been isolated from environments such as soil, water and plant tissues. Many strains are known for their capability of degrading aromatic molecules and producing extracellular polymers. A Gram-stain-negative, strictly aerobic, motile, red-pigmented, oxidase-negative, catalase-positive, rod-shaped strain, designated DH-S5 T , has been isolated from pork steak packed under CO 2 -enriched modified atmosphere. Cell diameters were 1.5×0.9 µm. Growth optima were at 30 °C and at pH 6.0. Phylogenetic analyses based on both complete 16S rRNA gene sequence and whole-genome sequence data revealed that strain DH-S5 T belongs to the genus Sphingomonas , being closely related to Sphingomonas alpina DSM 22537 T (97.4 % gene sequence similarity), followed by Sphingomonas qilianensis X1 T (97.4 %) and Sphingomonas hylomeconis GZJT-2 T (97.3 %). The DNA G+C content was 64.4 mol%. The digital DNA-DNA hybridization value between the isolate strain and S. alpina DSM 22537 T was 21.0 % with an average nucleotide identity value of 77.03 %. Strain DH-S5 T contained Q-10 as the ubiquinone and major fatty acids were C 18 : 1 cis 11 (39.3 %) and C 16 : 1 cis 9 (12.5 %), as well as C 16 : 0 (12.1 %) and C 14 : 0 2-OH (11.4 %). As for polar lipids, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, dimethylphosphatidylethanolamine and sphingoglycolipid could be detected, alongside traces of monomethylphosphatidylethanolamine. Based on its phenotypic, chemotaxonomic and phylogenetic characteristics, strain DH-S5 T (=DSM 110829 T =LMG 31606 T ) is classified as a representative of the genus Sphingomonas , for which the name Sphingomonas aliaeris sp. nov. is proposed.

Antimicrobial resistance (AMR) is a recognized global public health concern. Although the link between antimicrobial usage in food animals and AMR in humans is established, the detailed interactions are unclear. Antimicrobial stewardship (AMS) in livestock was first implemented in Europe with Sweden as the pioneer in 1986. Despite this head start, AMR is still an ongoing challenge for Europe. The European Union (EU) is an established agriculture producer, the second largest pork producer globally, and one of the largest markets for organic food. China is the global leader in both production and consumption of pork. China's rise in prosperity has led to an increase in its pork demand. Chinese producers commonly use antimicrobials during production for disease treatment and prevention to meet this increased demand. China's rising prosperity together with recent publicized food safety scandals, disease outbreaks in domestic livestock products, and increased AMR awareness have resulted in an increased willingness to pay and demand for organic food by Chinese consumers. Responding to the growing concerns of AMR by consumers and the World Health Organization (WHO), the Chinese government introduced a national pilot program in 2016 to reduce unnecessary antimicrobial use. Compared with China, the EU is a different entity as it is a political union comprising diverse countries and although it may have more experience in AMS, both entities face similar issues with AMR and increasing demand for organic food. Increased interest in organic food has arisen due to concerns about AMR, food safety, outbreaks of bacterial food contamination, and animal welfare. This article aims to compare the different AMS strategies employed by each entity, China and the EU, and how the increased demand for organic produce globally also influences the effort to reduce antimicrobial use in these entities' pork industries.

Rapid, sensitive and on-site monitoring of meat spoilage is highly essential for food safety. Hydrogen sulfide (H 2 S) a typical volatile, produced during enzymatic hydrolysis is considered as a reliable marker for evaluating meat freshness. Herein, a novel nano-bionic sensor based on the superior catalytic activity of ruthenium nanoparticles (Ru NPs) has been fabricated for H 2 S quantification. The activity sites of Ru NPs were poisoned in the presence of H 2 S, thereby affecting its catalytic efficiency via reducing the degradation of azo dye. The developed nano-bionic sensor achieved a selective response toward H 2 S, with capability for on-site surveillance of the pork freshness in the linear range (0-1800 nM). A higher correlation was obtained between the H 2 S content and the total viable count during the 9-period pork spoilage process (R 2  = 0.9633 and 0.9769). Moreover, the proposed method exhibits high selectivity in the presence of other characteristic volatiles encountered during the pork storage process., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

We present estimations for the amounts of Arcobacter (A. butzleri, A. cryaerophilus and A. skirrowii) and Campylobacter (C. jejuni, C. coli and C. fetus) species in retail chicken, pork and beef meat using PCR-MPN. Arcobacter butzleri, A. cryaerophilus and C. jejuni were found in 100, 60 and 55% of chicken samples, respectively. No other Arcobacter or Campylobacter species were found in chicken. The MPNs of A. butzleri, A. cryaerophilus and C. jejuni were greater than 10 3 per 100 g in 50, 0 and 5% of samples, respectively. The MPN of A. butzleri was higher than that of C. jejuni in 95% of samples. In pork, A. butzleri and A. cryaerophilus were detected in 10 and 11 (50 and 55%) of 20 samples, respectively. No other Arcobacter or Campylobacter species were found in pork. Only one pork sample had more than 10 3 MPN per 100 g of A. cryaerophilus. For beef, only two samples tested positive for A. cryaerophilus, at 4600 and 92 MPN per 100 g. Overall, we found that the presence and MPNs of Arcobacter species are very high in chicken. In contrast, the positive ratios of Arcobacter in pork were high as chicken samples, but MPNs were lower than in chicken., (© 2021 The Society for Applied Microbiology.)

Clostridium perfringens (C. perfringens), a prolific toxin-producing anaerobe is an important foodborne pathogen with a huge public health concern. Rapid and on-site detection of C. perfringens is of specific importance in developing countries. In the present study, saltatory rolling circle amplification (SRCA) assay was developed for culture-independent, rapid and visual detection of C. perfringens and evaluated in meat with pork as a model. The specificity of the SRCA assay was ascertained by using 62 C. perfringens and 18 non- C. perfringens strains. The analytical sensitivity of the developed SRCA, conventional and real-time PCR assays were 80 fg, 800 fg and 800 fg DNA per tube, respectively. The limit of detection of the SRCA assay was 80 CFU/g of pork in the absence of enrichment and 8 CFU/g after short enrichment of 6 h. The detection limits of 80 CFU/g and 8 CFU/g of pork were attained within 120 min and 8 h, respectively. Real-world or field relevancy of the developed assay was evaluated by screening 82 raw and processed pork samples. As the developed assay is simple, user-friendly, cost-effective and sophisticated-equipment free, it would be more suitable for on-site testing of C. perfringens in foods. To our information, this is the first report to apply SRCA for the detection of C. perfringens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Pork and pork products are important staple food in the diet of Vietnamese consumers. The safety of pork, including biological contamination, is a concern to several public authorities and value chain actors. This cross-sectional study aimed to identify Salmonella and total bacterial count (TBC) contamination of cut pork sold in different outlets, and determine the potential factors leading to contamination. A total of 671 pork samples were collected from different retail channels in three provinces in Northern Vietnam. Hygiene conditions and practices at pork vending premises were also observed and recorded. Data analysis used descriptive statistics and regression analysis. Overall, Salmonella prevalence in retailed pork was 58.1%. Salmonella contamination in pork from traditional retail, modern retail and food services were 60.5%, 50.9% and 80.5%, respectively. Eighty percent and 68% of fresh pork in canteen and street food was contaminated with Salmonella. Only a small proportion of a subset of the pork samples (6.2%) tested met the Vietnamese standard requirement for TBC contamination. Average concentration of TBC in fresh pork in traditional retail, modern retail and food services were 6.51 (SD: 0.64), 6.38 (0.65), and 6.96 (0.85) LogCFU/g, respectively. Transport time, use of the same tools for pork and other types of meat, storage temperature, and environment hygiene are important factors that might affect microbial contamination. The findings underline the high level of microbial contamination, which requires practical interventions to improve food safety hygiene practices and behavior of pork retailers., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)

This study aimed to investigate antimicrobial resistance (AMR) characteristics of Escherichia coli isolates from pig origin (including pigs, pig carcass, and pork) and humans in Thailand and Lao People's Democratic Republic (PDR) border provinces. The majority of the E. coli isolates from Thailand (69.7%) and Lao PDR (63.3%) exhibited multidrug resistance. Class 1 integrons with resistance gene cassettes were common ( n  = 43), of which the most predominant resistance gene cassette was aadA1 . The percentage of extended-spectrum beta-lactamase (ESBL) producers was 3.4 in Thailand and 3.2 in Lao PDR. The ESBL genes found were bla CTX-M14 , bla CTX-M27 , and bla CTX-M55 , of which bla CTX-M55 was the most common (58.6%). Ser-83-Leu and Asp-87-Asn were the predominant amino acid changes in GyrA of ciprofloxacin-resistant isolates. Twenty-two percent of all isolates were positive for qnrS . Class 1 integrons carrying aadA1 from pigs ( n  = 1) and ESBL genes ( bla CTX-M55 and bla CTX-M14 ) from pigs ( n  = 2), pork ( n  = 1), and humans ( n  = 7) were located on conjugative plasmids. Most plasmids (29.3%) were typed in the IncFrepB group. In conclusion, AMR E. coli are common in pig origin and humans in these areas. The findings confirm AMR as One Health issue, and highlight the need for comprehensive and unified collaborations within and between sectors on research and policy.

Yersinia enterocolitica bio-serotype 4/O:3 was previously identified in a pork production chain in Brazil and the obtained isolates presented high identity by pulsed-field gel electrophoresis (PFGE, XbaI). For the current study, an additional 147 porcine samples (tonsils = 100, palate = 30, head meat = 17) were collected from the same pork production chain 2-years later and 14 (9.5%) tested positive for Y. enterocolitica. Isolates (n = 24, 1 to 2 per positive sample) were bio-serotype 4/O:3 and harbored virulence genes ail, inv, wbbU, virF, myfA, ystA, ymoA, hreP and sat, and the multidrug resistance related genes emrD, marC and yfhD. PFGE (XbaI) demonstrated no differences among isolates (100% similarity) and were identical to some Y. enterocolitica isolates (n = 13) obtained previously from the same pork chain. A second PFGE analysis (NotI) confirmed the high degree of similarity among isolates obtained over time, demonstrating the persistence of an apparent clonal Y. enterocolitica bio-serotype 4/O:3 in this particular pork production chain in Brazil., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The aim of this study was to investigate the influence of the duration of frozen storage (12 and 24 weeks) and the storage temperature (-18 °C or - 80 °C) on the shelf life of pork in MAP for up to 14 days after thawing. Frozen meat was compared to unfrozen meat in different physicochemical and microbiological parameters. The parameters a*, b*, pH value, cooking loss, shear force and antioxidant activity showed no significant changes depending on the freezing process. The total moisture loss was significantly higher for frozen/thawed pork compared to unfrozen pork. The storage loss of all frozen samples showed higher values than the unfrozen samples on all examination days. The level of TBARS reached comparable values for all experimental groups. On examination days 7 and 14, significantly lower values of total plate count (TPC) and Pseudomonas spp. were measured for frozen pork compared to unfrozen pork. The temperature of frozen storage had no significant influence on the quality of pork., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Our purpose was to investigate the main bacterial microbiota and volatile profiles in the Chinese traditional dry-cured product-Jinhua ham during different processing stages and to analyze the role of the main microbiota in the formation of characteristic flavor. We determined the microbiota of Jinhua ham by using 16 S high throughput sequencing, and found that Staphylococcus constituted the predominant microbiota throughout the flavor formation process. Based on the volatile profiles of Jinhua dry-cured products from 11 different processing via SPME-GC-MS analysis, Aldehydes were the main groups of volatiles, with the most abundant ones being hexanal (13.89%) and nonanal (3.96%). To further investigate the relationship between predominant microbiota and the major volatile compounds in Jinhua ham, we screened and isolated genus Staphylococcus with high protease and lipase activities. The main Staphylococcus isolates, S. saprophyticus (53.4%) and S. equorum (31.0%) are related to the yields of aldehydes by producing hexanal, nonanal, benzaldehyde, and phenylacetaldehyde, indicating their contributions on the formation of characteristic flavor substances in Jinhua ham., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The emergence and dissemination of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli is a global health issue. Food-producing animals, including pigs, are significant reservoirs of antimicrobial resistance (AMR), which can be transmitted to humans. Thus, the rapid detection of ESBLs is required for efficient epidemiological control and treatment. In this study, multiplex recombinase polymerase amplification (RPA) combined with a single-stranded tag hybridization chromatographic printed-array strip (STH-PAS), as a lateral flow strip assay (LFA), was established for the rapid and simultaneous detection of multiple bla genes in a single reaction. Visible blue lines, indicating the presence of the blaCTX-M, blaSHV, and blaOXA genes, were observed within 10 min by the naked eye. The limit of detection of all three genes was 2.5 ng/25 μL, and no cross-reactivity with seven commensal aerobic bacteria was observed. A total of 93.9% (92/98) and 96% (48/50) of the E. coli isolates from pork meat and fecal samples, respectively, expressed an ESBL-producing phenotype. Nucleotide sequencing of the PCR amplicons showed that blaCTX-M was the most prevalent type (91.3-95.83%), of which the main form was blaCTX-M-55. The sensitivity and specificity of the RPA-LFA were 99.2% and 100%, respectively, and were in almost perfect agreement (κ = 0.949-1.000) with the results from PCR sequencing. Thus, the RPA-LFA is a promising tool for rapid and equipment-free ESBL detection and may facilitate clinical diagnosis in human and veterinary medicine, as well as AMR monitoring and surveillance., Competing Interests: The authors have declared that no competing interests exist.

The objective of this study was to develop an active banana starch film (BSF) incorporated with banana peel extract. We compared the film's properties with commercial wrap film (polyvinyl chloride; PVC). Moreover, a comparison of the quality of minced pork wrapped during refrigerated storage (7 days at ±4 °C) was also performed. The BSF with different concentrations of banana peel extract (0, 1, 3, and 5 (%, w/v )) showed low mechanical properties (tensile strength (TS): 4.43-31.20 MPa and elongation at break (EAB): 9.66-15.63%) and water vapor permeability (3.74-11.0 × 10 -10 g mm/sm 2 Pa). The BSF showed low film solubility (26-41%), but excellent barrier properties to UV light. The BSF had a thickness range of 0.030-0.047 mm, and color attributes were: L* = 49.6-51.1, a* = 0.21-0.43, b* = 1.26-1.49. The BSF incorporated with banana peel extracts 5 (%, w/v ) showed the highest radical scavenging activity (97.9%) and inhibitory activity of E. coli O157: H7. The BSF showed some properties comparable to the commercial PVC wrap film. Changes in qualities of minced pork were determined for 7 days during storage at ±4 °C. It was found that thiobarbituric acid reactive substances (TBARS) of the sample wrapped with the BSF decreased compared to that wrapped with the PVC. The successful inhibition of lipid oxidation in the minced pork was possible with the BSF. The BSF incorporated with banana peel extract could maintain the quality of minced pork in terms of oxidation retardation.

One objective of this study was to determine overall prevalence of Salmonella in ground pork from U.S. retail stores over three seasons including both case-ready and store-ground packages. Package types collected included: overwrap, chub, modified atmosphere packaging, and other (plastic or wax paper wrapped). Because package type represents different production systems and are subject to varied microbiological government regulation and testing methodologies, both USDA-FSIS and FDA Salmonella isolation protocols were performed. Another objective of the study was to determine serotypes and antimicrobial susceptibility profiles of the isolates obtained from the ground pork samples. Ground pork aliquots were subjected to real-time PCR. Recovered isolates were serotyped and minimum inhibitory concentration analysis to 15 antimicrobials was determined using microbroth dilution. Overall prevalence of Salmonella in ground pork from the 865 samples collected was 1.39%. Prevalence was not affected by package type ( p  = 0.29) nor grind location (case-ready vs. store-ground; p  = 0.17). Season affected Salmonella prevalence ( p  = 0.05) with most isolates found during fall, and there was a tendency for geographic region to affect prevalence ( p  = 0.07). The USDA Salmonella isolation method was more effective at recovering isolates ( p  = 0.01) compared with the FDA methodology and yielded a kappa statistic of 0.26 as a measure of agreement. The serotypes isolated included: Infantis, 4,5,12:i:-, Brandenburg, Typhimurium var 5-, Seftenberg, and Johannesburg with only two packages containing multiple serotypes. No isolates were resistant to antibiotics commonly used to treat human Salmonella infections including extended spectrum cephalosporins or fluoroquinolones. Although the recovery of Salmonella from retail ground pork samples was rare, Salmonella Typhimurium (and its monophasic variant 4,5,12:i:-), which are among the most common serovars recovered from human infections, were recovered. Therefore, more effective strategies to further reduce or eliminate these pathogens from retail pork products are warranted.