Modeling the growth of Salmonella in raw ground pork under dynamic conditions of temperature abuse.

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 (µmax) 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 µmax 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.

Functional genome analysis and anti-Helicobacter pylori activity of a novel bacteriocinogenic Lactococcus sp. NH2-7C from Thai fermented pork (Nham).

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 5805T. 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.

Evaluación de la resistencia a los antibióticos de cepas de Escherichia coli aisladas en carne de cerdo comercializada en los mercados municipales de la ciudad de Guatemala / Evaluation of antibiotics resistance of Escherichia coli strains isolated in pork sold in municipal markets of Guatemala City

La resistencia a los antimicrobianos es un problema de salud pública a nivel mundial que va en aumento y se ve reflejada en la falta de eficacia de los tratamientos de infecciones bacterianas con antibióticos en humanos y en animales. El presente estudio tuvo como objetivo evaluar la resistencia a los antibióticos de cepas de Escherichia coli aisladas en carne de cerdo expendida en los mercados municipales de la ciudad de Guatemala. Se identificaron los antibióticos que presentaron mayor resistencia y mayor sensibilidad in vitro frente a las cepas de E. coli aisladas a partir de 76 muestras de carne de cerdo. Se realizó un muestreo aleatorio simple con afijación proporcional por mercado. Para la identificación de las cepas de E. coli se utilizó la prueba de IMViC y para evaluar la resistencia a los antimicrobianos se utilizó la prueba de Kirby Bauer empleando 9 antibióticos. Se aisló E. coli en el 55% (42/76) de las muestras. La resistencia en las 42 cepas aisladas fue: tetraciclina (83%) neomicina (50%) y sulfametoxasole + trimetoprim (50%). 83% de las cepas (35/42) fueron resistentes a 2 antibióticos y 50% (21/42) a 3 antibióticos o más. Se obtuvo mayor sensibilidad con ceftriaxona (91%), amikacina (83%), gentamicina (65%) y ácido nalidíxico (65%). Se concluye que existe resistencia a los antibióticos evaluados, lo que constituye un riesgo para la salud pública ya que se encuentra en cepas aisladas en un alimento para consumo humano.

Antimicrobial resistance is a global public health threat that is increasing and is reflected in the lack of efficacy of bacterial infection treatments with antibiotics in humans and animals. The objective of this study was to evaluate the resistance to antibiotics of Escherichia coli strains isolated from pork in the municipal markets of Guatemala City. Antibiotics with the highest resistance and those with the highest sensitivity in vitro against the strains of E. coli were evaluated. A simple random sampling was carried out with proportional allocation by market, and 76 samples were collected. IMViC test was used to identify the E. coli strains, and antibiotics resistance was evaluated using the Kirby Bauer with nine different antibiotics. E. coli was isolated in 55% (42/76) of the samples. Resistance was evaluated in the 42 isolates. Antibiotic resistance was detected to tetracycline (83%), neomycin (50%), and sulfamethoxazole + trimethoprim (50%). All isolates presented resistance to at least one antibiotic; it was determined that 83% (35/42) showed resistance to two antibiotics and 50% (21/42) showed resistance to three antibiotics or more. The sensitivity obtained was higher for ceftriaxone (91%), amikacin (83%), gentamicin (65%), and nalidixic acid (65%). In conclusion, antibiotic resistance was detected, which constitutes a risk to public health since it is found in isolated strains in food for human consumption.

Effect of Morus alba leaf extract dose on lipid oxidation, microbiological stability, and sensory evaluation of functional liver pâtés during refrigerated storage.

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.

Evaluation of the Membrane Damage Mechanism of Chlorogenic Acid against Yersinia enterocolitica and Enterobacter sakazakii and Its Application in the Preservation of Raw Pork and Skim Milk.

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.

Application of gas sensors for modelling the dynamic growth of Pseudomonas in pork stored at different temperatures.

Pseudomonas have a faster growth rate over other bacteria in chilled meat under aerobic conditions. A non-destructive method for modelling the dynamic growth of Pseudomonas in pork stored at different temperatures using gas sensors was presented in our work. Based on selected gas sensor data, the first-order kinetic equations (Gompertz and Logistic Functions) combined with the secondary model (Square-root Function) effectively simulated Pseudomonas growth in pork at different temperatures with R2 and RMSE values of 0.71-0.97 and 0.27-0.84, respectively. Additionally, these models showed high accuracy with correlation coefficients greater than 0.90, in addition to several individual accuracy values. Furthermore, HS-SPME/GC-MS results demonstrated the presence of identified key volatiles in samples inoculated with Pseudomonas, including three amine compounds (mercaptamine, 1-octanamine and 1-heptadecanamine), phenol and indole. Our work showed that gas sensors are a rapid, easy and non-destructive method with acceptable feasibility in modelling the dynamic growth of spoilage microorganisms in meat.

Investigation of Listeria monocytogenes, Salmonella enterica and Yersinia enterocolitica in pig carcasses in Southern Brazil / Pesquisa de Listeria monocytogenes, Salmonella enterica and Yersinia enterocolitica em carcaças de suínos no sul do Brasil

The intensification of pig production and advances in the sanitary control of herds profoundly changed the profile of risk attributed to pork consumption. In the actual scenario, most microorganisms related to macroscopic lesions observed in the post mortem inspection are not transmitted by food, while foodborne bacteria of importance to consumer health do not cause macroscopic lesions. In Brazil, the "Ministério da Agricultura, Pecuária e Abastecimento" requested a scientific opinion on the prioritizing of pathogens potentially transmitted by unprocessed pork. After conducting a qualitative risk assessment, only Salmonella enterica was classified as of high risk to consumers. The present study was part of the validation step of the risk assessment and aimed to investigate the frequency of S. enterica, Yersinia enterocolitica and Listeria monocytogenes and hygienic-sanitary indicators in pig carcasses of pigs rose under intensive production and slaughtered under the Federal Inspection System in three slaughterhouses located in Southern Brazil. Additionally, the antimicrobial resistance profile of the isolated pathogens was also investigated. A total of 378 carcasses were sampled by superficial sponges before the chilling step in three slaughterhouses. Samples were investigated for the presence of the three aforementioned pathogens and subjected to enumeration of Colony Formation Units (log CFU.cm-1) of total aerobic mesophiles (TAM) and Enterobacteriaceae. Salmonella strains were tested by disc diffusion test for resistance to eleven antimicrobials. There were significantly statistical differences (p<0.0001) on the median counts of both indicators between the slaughterhouses. The median of TAM was very close for Slaughterhouses A and B: 1.573 log CFU.cm-1 and 1.6014 log CFU.cm-1, respectively. While in Slaughterhouse C, a higher TAM median was detected (2.216 log CFU.cm-1). A similar profile was observed regarding to Enterobacteriaceae, and medians were calculated as follow: -0.426 log CFU.cm-1 in Slaughterhouse A; 0.2163 log CFU.cm-1 in B; and 0.633 log CFU.cm-1 in C. Regarding the pathogens investigated, L. monocytogenes was not detected and only one carcass from Slaughterhouse C was positive for Y. enterocolitica. Thus, the results suggest a very low prevalence of L. monocytogenes and Y. enterocolitica in the sampled population. A total of 65 (17.2%) carcasses were positive for S. enterica, with a difference in frequencies between slaughterhouses and slaughter days. The prevalence of Salmonella positive carcasses was higher in the Slaughterhouse C (25.4%; CI 95% 19-32%) in comparison with A (9.5%; CI 95% 9-14%) and B (18.3%; CI 95% 12-24%). There was no significantly statistical association between Enterobacteriaceae counts and Salmonella isolation on carcass surface (p=0.69). The slaughtering day, nested within the slaughterhouse, explains 31.3% of Salmonella prevalence variability. S. Typhimurium (38.1%) was the most prevalent, followed by S. Infantis (30.1%). Among the 61 Salmonella strains tested for resistance to antimicrobials, 18 (31.6%) were full-susceptible. No strain displayed resistance to azithromycin, ceftazidime, cefotaxime and meropenem. The highest resistance frequency was displayed to tetracycline (54.1%), followed by ampicillin (50.82%), nalidixic acid (42.62%) and chloramphenicol (42.62). Multi-resistance was detected in 52.54% of the, strains. In conclusion, S. enterica is more prevalent in pre-chill pig carcasses than Y. enterocolitica and L. monocytogenes and thus should be prioritized in monitoring and control programs at slaughter. Salmonella serovars varied among slaughterhouses and present significant differences in their resistance to antimicrobials. Slaughterhouses that present higher medians of TAM or Enterobacteriaceae in a monitoring period may have higher S. enterica prevalences as well. However, there is a high variation of S. enterica prevalence among slaughter days, which cannot be always related to the hygienic indicators counts observed on a given day.(AU)

A intensificação da produção de suínos e os avanços no controle sanitário dos rebanhos alterou de forma importante o perfil de risco do consumo de carne suína. No cenário atual, a maioria dos microrganismos causadores de lesões macroscópicas detectáveis na inspeção post mortem não são transmissíveis por alimentos, enquanto bactérias de importância como causadoras de doenças transmitidas por alimentos não causam lesões macroscópicas. No Brasil, o Ministério da Agricultura, Pecuária e Abastecimento solicitou uma opinião científica sobre a priorização de patógenos potencialmente transmitidos pela carne suína in natura. Após conduzir uma avaliação de risco qualitativa, apenas Salmonella enterica foi classificada como de alto risco para o consumidor. O presente estudo foi parte da etapa de validação da avaliação de risco e objetivou: investigar a frequência de S. enterica, Yersinia enterocolitica e Listeria. monocytogenes; e enumerar indicadores higiênico-sanitários em carcaças de suínos abatidos sob inspeção federal em frigoríficos dedicados ao abate de suínos sob sistema intensivo de criação no sul do Brasil. Além disso, o perfil de resistência a antimicrobianos dos patógenos isolados foi investigado. A superfície de um total de 378 carcaças foi amostrada por esponjas, na etapa de pré-resfriamento em três matadouros frigoríficos (A, B, C). As amostras foram investigadas quanto à presença dos três patógenos acima mencionados e quanto à enumeração de Unidades Formadoras de Colônia (log UFC.cm-1) de mesófilos aeróbios totais (MAT) e Enterobacteriaceae. As cepas isoladas de Salmonella foram testadas quanto à resistência a onze antimicrobianos pela técnica de disco difusão. As medianas de contagem de ambos os indicadores apresentaram diferença significativa (p<0,0001) entre matadouros-frigoríficos. A mediana de MAT foi bastante próxima para A e B (1,573 log UFC.cm-1 e 1,6014 log UFC.cm-1, respectivamente), enquanto em C uma mediana de MAT mais elevada foi determinada (2,216 log CFU.cm-1). Um perfil semelhante foi observado em relação a Enterobacteriaceae, sendo as medianas calculadas para A, B e C, respectivamente: -0,426 log CFU.cm-1; 0,2163 log UFC.cm-1; e 0,633 log UFC.cm-1. Em relação aos patógenos investigados, L. monocytogenes não foi detectada e apenas uma carcaça, do Matadouro C, foi positiva para Y. enterocolitica. Portanto, os resultados sugerem uma prevalência muito baixa desses patógenos na população amostrada. Em um total de 65 (17,2%) carcaças houve isolamento de S. enterica, com diferença nas frequências observadas entre matadouros e dias de abate. A prevalência de carcaças positivas para S. enterica foi maior no Matadouro C (25,4%; IC95% 19-32%) em comparação com A (9,5%; IC95% 9-14%) e B (18,3%; IC95% 12-24%). Não houve associação estatística entre o número de Enterobacteriaceae e o isolamento de S. enterica na superfície das carcaças (p=0,69). O dia de abate agrupado por frigorífico explica 31,3% da variação na prevalência de Salmonella. O sorovar mais frequente de S. enterica foi Typhimurium (38,1%) seguido de S. Infantis (30,1%). Entre as 61 cepas de S. enterica testadas quanto à resistência a antimicrobianos, 18 (31,6%) foram totalmente suscetíveis aos antimicrobianos testados. Nenhuma cepa apresentou resistência a azitromicina, ceftazidima, cefotaxima e meropenem. As maiores frequências de resistência foram demonstradas contra tetraciclina (54,1%), ampicilina (50,8%), ácido nalidíxico (42,62%) e cloranfenicol (42,62%). Em 52,54% das cepas foi detectada multi-resistência. Em conclusão, S. enterica é mais prevalente em carcaças suínas no pré-resfriamento do que Y. enterocolitica e L. monocytogenes. Portanto, S. enterica deve ser priorizada em programas de monitoramento e controle ao abate. Os sorovares de Salmonella variam entre matadouros e apresentam diferenças significativas na resistência a antimicrobianos. Matadouros de suínos que apresentam medianas de MAT e Enterobacteriaceae num período de monitoramento podem apresentar também prevalências mais de altas de presença de S. enterica. Entretanto, há uma alta variabilidade na frequência de S. enterica entre dias de abate, e nem sempre há relação entre essa frequência e a contagem de indicadores higiênico-sanitários determinados num determinado dia.(AU)

Effect of chitosan-gelatin coating containing nano-encapsulated tarragon essential oil on the preservation of pork slices.

There is an increasing demand for bio-based packaging materials incorporated with active nanoparticles as one of the new technologies to obtain food products with improved quality and extended shelf-life. The objectives of this study were to develop a chitosan-gelatin based edible coating incorporated with nano-encapsulated tarragon essential oils (TEO) and to investigate the effects of chitosan-gelatin coatings containing TEO or TEO-loaded nanoparticles (TEO-NPs) on the preservation of pork slices during refrigerated storage for 16 days. Nanoparticles with the average diameters of 246.27-504.60 nm were produced using different mass ratios of chitosan to TEO (1:0, 1:0.2, 1:0.4, 1:0.6, 1:0.8 and 1:1) by ionic gelation method. Zeta potential and encapsulation efficiency (EE) of the prepared TEO-NPs were 27.07-37.12 mV and 9.83-35.57%, respectively. TEO-NPs with the maximum EE were applied for preparing the edible coatings. The results suggested that the coating treatments could significantly inhibit quality deterioration of pork slices. Nano-encapsulation contributed to the sustained release of TEO and caused an improved antioxidant, antibacterial and sensory properties. The study suggests that chitosan-gelatin coating incorporated with TEO-NPs could be developed as a prospective active packaging to preserve pork slices.

Fatal case of bacteremia caused by Streptococcus suis in a splenectomized man and a review of the European literature.

Streptococcus suis is an emerging zoonotic human pathogen, which is a causative agent of invasive infections in people who are in close contact with infected pigs or contaminated pork products. It is associated with severe systemic infections, most commonly meningitis and sepsis, which may lead to high rates of morbidity and mortality. Serotype 2 is the most prevalent type in S. suis infections in humans. We have reported a case of a very rapidly proceeding fatal human S. suis infection in a splenectomized, but otherwise immunocompetent patient in Hungary. We would like to highlight the attention for this pathogen for the risk group patients, not only pig breeders, veterinarians, abattoir workers, meat processing and transport workers, butchers and cooks, that those persons who are immunocompromised including those with spleen removed, persons with diabetes mellitus, cancer and alcoholism, are also at greater risk of infection.

An FcεRI-IgE-based genetically encoded microfluidic cell sensor for fast Gram-negative bacterial screening in food samples.

An FcεRI-IgE-based genetically encoded microfluidic cell sensor was constructed for fast Gram-negative bacterial screening in food samples. CD14-Fcε IgE, produced by the gene engineered antibodies (GEAs) technology, was used for the recognition of the target bacteria or lipopolysaccharide (LPS). Stable cell lines expressing GCaMP6s, a genetically encoded indicator of calcium flux, were first established for monitoring mast cell activation and improving detection sensitivity. The microfluidic system was designed to improve automation and control the reaction time. Once Gram-negative bacteria bound to the CD14-Fcε IgE on the RBL-2H3 cell surface, RBL-2H3 cell receptor (FcεRI)-induced Ca2+ signaling pathway was immediately activated to release Ca2+. The elevated intracellular Ca2+ triggers GCaMP6s for reporting the presence of Gram-negative bacteria. The developed biosensor was able to detect 80 CFU mL-1 Gram-negative bacteria within 2.5 min in pure culture samples. The biosensor was used to detect Gram-negative bacteria in pork samples. With its short screening time and easy operation, the proposed biosensor shows promise in future applications of foodborne pathogen testing in 1 h to 1 day.

The effect of barrier properties of polymeric films on the shelf-life of vacuum packaged fresh pork meat.

The aim of this study was to investigate the permeability properties of synthetic polymeric materials and their influence on the quality of packed chilled pork. Barrier properties of biaxially oriented polypropylene (BOPP) coated with acrylic/polyvinylidene chloride (BOPPAcPVDC) and biaxially oriented coextruded polypropylene (BOPPcoex), were tested on three gases: oxygen, carbon dioxide and nitrogen at different temperatures: 4 °C, 20 °C, 40 °C and 60 °C. Coefficients of permeability, diffusion and solubility were determined. The quality parameters of vacuum packed fresh chilled pork stored under controlled temperature over a 21-day period were evaluated with physico-chemical, sensory and microbiological analyses. Results showed that BOPPAcPVDC film provided better protection for fresh pork than BOPPcoex film in terms of physico-chemical, sensory and microbiological attributes, due to its greater barrier behaviour, with smaller permeance data (q) (0.98 to 324 cm3/m2 · d · bar) for BOPPAcPVDC, compared to BOPPcoex (227 to 6200 cm3/m2 · d · bar), in the temperature range from 4 °C to 60 °C (p < .001). The shelf life of fresh pork packed in this film was doubled, from 7 to 14 days.