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Introduction: Salmonellosis is the second most commonly occurring bacterial zoonosis in Germany. Rye in pig feeding offers new possibilities for addressing that issue due to its high content of non-starch polysaccharides (NSPs). These are fermented in the intestinal tract to specific fermentation products, which seem to have bacteriolytic effects against Salmonella. A coarse feed structure can display synergistic effects., Methods: Seven conventional pig fattening farms increased the rye content (40%-70%) while limiting the amount of fine particles (maximum of 20% ≤0.25 mm). Samples from pigs were tested for Salmonella antibodies and compared with samples from 167 farms without any changes to the feed., Results: Rye-based diets had a significant (p < 0.05) impact on Salmonella antibody (percentage optical density [OD%]) detection. In this study, it became apparent that significantly fewer positive OD% values could be detected due to the increase in rye compared to farms that did not change the feed (Farm 6 P0: 35.45 ± 36.18; P1: 15.48 ± 16.98; P2: 9.36 ± 8.17). An elimination of Salmonella could not be achieved, but especially farms with high antibody counts were able to strongly reduce those in both phases consecutively (Farm 5 P0: 35.17 ± 35.53; P1: 18.56 a  ± 20.96; P2: 13.38 a  ± 18.99). That was different on farms without adapted feeding, where an increase in Salmonella antibodies was observed (P0: 17.38 ± 22.21; P1: 20.12 ± 25.39; P2: 18.12 ± 25.44)., Conclusion: By increasing the proportion of rye and limiting the proportion of fine particles in the feed, Salmonella antibodies (OD% values) in meat juice and blood can be significantly reduced, especially on farms with an initially high incidence of Salmonella. If that is implemented in feeding across the board on farms, an improvement in food safety and a decreased risk of zoonosis can be expected., (© 2024 The Author(s). Veterinary Medicine and Science published by John Wiley & Sons Ltd.)

In Gram-negative bacteria, LPS (lipopolysaccharide) has been thoroughly characterized and has been shown to play a major role in pathogenesis and bacterial defense. In Salmonella and Escherichia coli , LPS also influences biofilm development. However, the overall role of LPS glycoform in biofilm formation has not been conclusively settled, as there is a lack of consensus on the topic. Some studies show that LPS mutants produce less biofilm biomass than the wild-type strains, while others show that they produce more. This review summarizes current knowledge of LPS biosynthesis and explores the impact of defective steps on biofilm-related characteristics, such as motility, adhesion, auto-aggregation, and biomass production in Salmonella and E. coli . Overall, motility tends to decrease, while adhesion and auto-aggregation phenotypes tend to increase in most LPS-mutant strains. Interestingly, biofilm biomass of various LPS mutants revealed a clear pattern dependent on biofilm maturation time. Incubation times of less than 24 h resulted in a biofilm-defective phenotype compared to the wild-type, while incubation exceeding 24 h led to significantly higher levels of biofilm production. This explains conflicting results found in reports describing the same LPS mutations. It is therefore critical to consider the effect of biofilm maturation time to ascertain the effects of LPS glycoform on biofilm phenotype. Underlying reasons for such changes in biofilm kinetics may include changes in signalling systems affecting biofilm maturation and composition, and dynamic LPS modifications. A better understanding of the role of LPS in the evolution and modification of biofilms is crucial for developing strategies to disperse biofilms., Competing Interests: The authors declare no conflict of interest.

Salmonella forms biofilms, and persist on food contact surfaces. Once a biofilm is formed cleaning and sanitation protocols may be inadequate for effective removal. This study evaluated attachment characteristics, surface properties, and structure of Salmonella biofilms on food contact surfaces commonly used in the tree-fruit industry. Multi-strain Salmonella biofilms were grown in a Centers for Disease Control and Prevention (CDC) biofilm reactor at 22 ± 2 °C and sampling was conducted at 2, 24 and 96-h. After each incubation period, coupons weregently rinsed and the remaining cells enumerated. Biofilms were analyzed with Laser Scanning Confocal Microscopy (LSCM). Hydrophobicity was evaluated by measuring the contact angles of reference liquids method using a drop tensiometer instrument. Material type and biofilm age significantly influenced attachment and biofilm hydrophobicity (P < 0.05). The strength of attachment, across all time points, was highest on nylon followed by wood and high-density polyethylene. The highest contact angle measurements were observed after 96-h of biofilm formation for each material. All the results and observations from this study contribute to a better understanding of the attachment and hydrophobicity characteristics of Salmonella and might help producers make informed decisions when selecting containers for harvesting and storing in order to minimize biofilm formation and potential for cross-contamination., (© 2024. The Author(s).)

Probiotics are increasingly recognized for their capacity to combat pathogenic bacteria. In this study, we isolated a strain of Ligilactobacillus salivarius XP132 from the gut microbiota of healthy chickens. This strain exhibited resistance to low pH and bile salts, auto-aggregation capabilities, and the ability to co-aggregate with pathogenic Salmonella. The in vitro antibacterial activity of Ligilactobacillus salivarius XP132 was tested using an Oxford cup antibacterial test, and the results showed that Ligilactobacillus salivarius XP132 exhibited broad-spectrum antibacterial activity, with especially strong antibacterial activity against Salmonella. In animal experiments with white feather broilers and specific-pathogens-free (SPF) chickens, we orally administered 1 × 10 9 CFU XP132 live bacteria per chicken per day, and detected the content of Salmonella in the liver, spleen, intestinal contents, and eggs of the chickens by RT-qPCR. Oral administration of Lactobacillus salivarius XP132 group significantly reduced the levels of Salmonella in chicken liver, spleen, intestinal contents and eggs, and the oral administration of Ligilactobacillus salivarius XP132 significantly inhibited the horizontal and vertical transmission of Salmonella in SPF chickens and white-feathered broilers. After oral administration of XP132, the production of chicken serum anti-infective cytokine IFN-γ was also significantly up-regulated, thereby enhancing the host's ability to resist infection. In addition, the production of various serum inflammatory cytokines, including IL-1β, IL-6, IL-8, and TNF-α, was down-regulated, leading to significant amelioration of the inflammatory response induced by S. Pullorum in chickens. These findings suggest that Ligilactobacillus salivarius XP132 possesses potent antibacterial and immunomodulatory properties that effectively prevent both horizontal and vertical transmission of Salmonella Pullorum, highlighting its potential as a valuable tool for the prevention and control of Salmonella disease., Competing Interests: DISCLOSURES The authors declare no conflict of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonella and Campylobacter are major foodborne pathogens that cause outbreaks associated with contaminated chicken liver. Proper cooking is necessary to avoid the risk of illness to consumers. This study tested the thermal inactivation of a 4-strain Salmonella cocktail and a 3-strain Campylobacter cocktail in chicken livers separately at temperatures ranging from 55.0 to 62.5°C. Inoculated livers were sealed in aluminum cells and immersed in a water bath. The decimal reduction time (D-values) of Salmonella in chicken livers were 9.01, 2.36, 0.82, and 0.23 min at 55.0, 57.5, 60.0, and 62.5°C, respectively. The D-values of Campylobacter ranged from 2.22 min at 55.0°C to 0.19 min at 60.0°C. Salmonella and Campylobacter had similar z-values in chicken livers of 4.8 and 4.6°C, respectively. Chicken livers can be heated to internal temperatures of 70.0 to 73.9°C for at least 1.6 to 0.2 s to achieve a 7-log reduction of Salmonella. Validation tests demonstrated that heating chicken livers to internal temperatures of 70.0 to 73.9°C for 2 to 0 s resulted in a reduction of Salmonella exceeding 7 logs. Collectively, these data show that Salmonella exhibits higher heat resistance than Campylobacter in chicken livers. Therefore, Salmonella could be considered as the target pathogen when designing thermal treatments or cooking instructions for liver products. These findings will aid in designing effective thermal processing for both industrial and home cooking to eliminate Salmonella and Campylobacter, ensuring consumer safety when consuming chicken liver products., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

1. Epidemiological surveillance of Salmonella spp. serves as a primary tool for maintaining the health of poultry flocks. Characterising circulating serotypes is crucial for implementing control and prevention measures. This study conducted phenotypic and molecular characterisation of S. enterica Pullorum, S. enterica Heidelberg, and S. enterica Corvalis isolated from broiler chickens during slaughtering.2. All strains were susceptible to gentamicin, neomycin and norfloxacin. However, resistance rates exceeded 50% for ciprofloxacin and tiamulin, irrespective of the serotype. Approximately 64% of strains were classified as multidrug-resistant, with S. enterica Heidelberg strains exhibiting significantly higher overall resistance. The isolates demonstrated the ability to adhere and produce biofilm at a minimum of three temperatures, with S. enterica Pullorum capable of biofilm production at all temperatures encountered during poultry rearing.3. Each strain possessed between two and seven different virulence-associated genes. Genetic similarity, as indicated by pulsed field gel electrophoresis, exceeded 90% for all three serotypes and strains were classified in the R5 ribotype by PCR, regardless of serotype. Sequencing revealed high similarity among all strains, with homology ranging from 99.61 to 100% and all were classified to a single cluster.4. The results suggested a clonal relationship among the strains, indicating the possible circulation of a unique clonal group of S. enterica Pullorum in the southern region of Brazil.

Microbial thermal inactivation in low moisture foods is challenging due to enhanced thermal resistance of microbes and low thermal conductivity of food matrices. In this study, we leveraged the body of previous work on this topic to model key experimental features that determine microbial thermal inactivation in low moisture foods. We identified 27 studies which contained 782 mean D-values and developed linear mixed-effect models to assess the effect of microorganism type, matrix structure and composition, water activity, temperature, and inoculation and recovery methods on cell death kinetics. Intraclass correlation statistics (I 2 ) and conditional R 2 values of the linear mixed effects models were: E. coli (R 2 -0.91, I 2 -83%), fungi (R 2 -0.88, I 2 -85%), L. monocytogenes (R 2 -0.84, I 2 -75%), Salmonella (R 2 -0.69, I 2 -46%). Finally, global response surface models (RSM) were developed to further study the non-linear effect of a w and temperature on inactivation. The fit of these models varied by organisms from R 2 0.88 (E. coli) to 0.35 (fungi). Further dividing the Salmonella data into individual RSM models based on matrix structure improved model fit to R 2 0.90 (paste-like products) and 0.48 (powder-like products). This indicates a negative relationship between data diversity and model performance., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Transfer of Salmonella to internal organs of broilers over a 35 d grow-out period was evaluated. A total of 360 one-day old chicks were placed in 18 floor pens of 3 groups with 6 replicate pens each. On d 0, broilers were orally challenged with a cocktail of Salmonella (equal population of marked serovars; nalidixic acid-resistant S. Typhimurium, rifampicin-resistant S. Infantis, and kanamycin-resistant S. Reading) to have 3 groups: L (low; ∼2 log CFU/bird); M (medium; ∼5 log CFU/bird); and H (High; ∼8 log CFU/bird). On d 2, 7 and 35, 4 birds/pen were euthanized and ceca, liver, and spleen samples were collected aseptically. Gizzard samples (4/pen) were collected on d 35. The concentration of Salmonella in liver and spleen were transformed to binary outcomes (positive and negative) and fitted in glm function of R using cecal Salmonella concentrations (log CFU/g) and inoculation doses (L, M, and H) as inputs. On d 2, H group showed greater (P ≤ 0.05) cecal colonization of all 3 serovars compared to L and M groups. However, M group showed greater (P ≤ 0.05) colonization of all 3 serovars in the liver and spleen compared to L group. Salmonella colonization increased linearly in the ceca and quadratically in the liver and spleen with increasing challenge dose (P ≤ 0.05). On d 35, L group had greater (P ≤ 0.05) S. Infantis colonization in the ceca and liver compared to M and H groups (P ≤ 0.05). Moreover, within each group on d 35, the concentration of S. Reading was greater than those of S. Typhimurium and S. Infantis for all 3 doses in the ceca and high dose in the liver and gizzard (P ≤ 0.05). Salmonella colonization diminished in the ceca, liver, and spleen during grow-out from d 0 to d 35 (P ≤ 0.05). On d 35, birds challenged with different doses of Salmonella cocktail showed a similar total Salmonella spp. population in the ceca (ca. 3.14 log CFU/g), liver (ca. 0.54 log CFU/g), spleen (ca. 0.31 log CFU/g), and gizzard (ca. 0.42 log CFU/g). Estimates from the fitted logistic model showed that one log CFU/g increase in cecal Salmonella concentration will result in an increase in relative risk of liver and spleen being Salmonella-positive by 4.02 and 3.40 times (P ≤ 0.01), respectively. Broilers from H or M group had a lower risk (28 and 23%) of being Salmonella-positive in the liver compared to the L group when the cecal Salmonella concentration is the same (P ≤ 0.05). Oral challenge of broilers with Salmonella spp. with various doses resulted in linear or quadratic increases in Salmonella colonization in the internal organs during early age and these populations decreased during grow-out (d 35). This research can provide guidance on practices to effectively mitigate the risk of Salmonella from chicken parts and enhance public health., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonellosis, one of the most common foodborne infections in Europe, is monitored by food safety surveillance programmes, resulting in the generation of extensive databases. By leveraging tree-based machine learning (ML) algorithms, we exploited data from food safety audits to predict spatiotemporal patterns of salmonellosis in northwestern Italy. Data on human cases confirmed in 2015-2018 (n = 1969) and food surveillance data collected in 2014-2018 were used to develop ML algorithms. We integrated the monthly municipal human incidence with 27 potential predictors, including the observed prevalence of Salmonella in food. We applied the tree regression, random forest and gradient boosting algorithms considering different scenarios and evaluated their predictivity in terms of the mean absolute percentage error (MAPE) and R 2 . Using a similar dataset from the year 2019, spatiotemporal predictions and their relative sensitivities and specificities were obtained. Random forest and gradient boosting (R 2  = 0.55, MAPE = 7.5%) outperformed the tree regression algorithm (R 2  = 0.42, MAPE = 8.8%). Salmonella prevalence in food; spatial features; and monitoring efforts in ready-to-eat milk, fruits and vegetables, and pig meat products contributed the most to the models' predictivity, reducing the variance by 90.5%. Conversely, the number of positive samples obtained for specific food matrices minimally influenced the predictions (2.9%). Spatiotemporal predictions for 2019 showed sensitivity and specificity levels of 46.5% (due to the lack of some infection hotspots) and 78.5%, respectively. This study demonstrates the added value of integrating data from human and veterinary health services to develop predictive models of human salmonellosis occurrence, providing early warnings useful for mitigating foodborne disease impacts on public health., (© 2024. The Author(s).)

Plant extracts are increasingly recognized as potential prophylactic agents in poultry production due to their diverse bioactive properties. This study investigated the phytochemical and biological properties of Libidibia ferrea (L. ferrea), a plant species native to the Caatinga region of northeastern Brazil. The aim of this study was to identify secondary metabolites and to demonstrate the antimicrobial, antioxidant and protective effects of the plant extract. Three extracts were produced: EHMV, a hydroalcoholic extract from the maceration of pods, and EEMC and EEMV ethanolic extracts from the maceration of peels and pods, respectively, from L. ferrea. High-performance liquid chromatography (HPLC-MS/MS) and atomic absorption spectroscopy (AAS) were used to characterize the metabolites and metals. The antimicrobial activity against Salmonella Galinarum (SG), Salmonella pullorum (SP), Salmonella Heidelberg (SH) and Avian pathogenic Escherichia coli (APEC) was evaluated alone and in combination with probiotic bacteria (Bacillus velenzensis) using agar diffusion and the bactericidal minimum concentration (CBM). The antioxidant potential of the extracts was evaluated in 5 in vitro assays and 6 assays in 3t3 cells. The toxicity of EHMV was tested, and its ability to combat SP infection was demonstrated using a chicken embryo model. The results showed that EHMV exhibited significant antimicrobial activity. The combination of EHMV with BV had synergistic effects, increased antimicrobial activity and induced bacterial sporulation. Composition analysis revealed the presence of 8 compounds, including tannins and phenolic compounds. In vitro antioxidant tests demonstrated that total antioxidant capacity(TAC) activity was increased, and the extract had strong reducing power and notable metal chelating effects. Analysis of 3T3 cells confirmed the protective effect of EHMV against oxidative stress. Toxicity assessments in chicken embryos confirmed the safety of EHMV and its protective effect against SP-induced mortality. EHMV from L. ferrea is rich in proteins and contains essential metabolites that contribute to its antimicrobial and antioxidant properties. When associated with probiotic bacteria such as B. velezensis, this extract increases the inhibition of SH, SG, SP, and APE. The nontoxic nature of EHMV and its protective effects on chicken embryos make it a potential supplement for poultry., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Salmonellosis, caused by Salmonella species, is one of the most common foodborne illnesses worldwide with an estimated 93.8 million cases and about 155,00 fatalities. In both industrialized and developing nations, Salmonellosis has been reported to be one of the most prevalent foodborne zoonoses and is linked with arrays of illness syndromes such as acute and chronic enteritis, and septicaemia. The two major and most common Salmonella species implicated in both warm-blooded and cold-blooded animals are Salmonella bongori and Salmonella enterica. To date, more than 2400 S. enterica serovars which affect both humans and animals have been identified. Salmonella is further classified into serotypes based on three primary antigenic determinants: somatic (O), flagella (H), and capsular (K). The capacity of nearly all Salmonella species to infect, multiply, and survive in human host cells with the aid of their pathogenic and virulence arsenals makes them deadly and important public health pathogens. Primarily, food-producing animals such as poultry, swine, cattle, and their products have been identified as important sources of salmonellosis. Additionally, raw fruits and vegetables are among other food types that have been linked to the spread of Salmonella spp. Based on the clinical manifestation of human salmonellosis, Salmonella strains can be categorized as either non-typhoidal Salmonella (NTS) and typhoidal Salmonella . The detection of aseptically collected Salmonella in necropsies, environmental samples, feedstuffs, rectal swabs, and food products serves as the basis for diagnosis. In developing nations, typhoid fever due to Salmonella Typhi typically results in the death of 5%-30% of those affected. The World Health Organization (WHO) calculated that there are between 16 and 17 million typhoid cases worldwide each year, with scaring 600,000 deaths as a result. The contagiousness of a Salmonella outbreak depends on the bacterial strain, serovar, growth environment, and host susceptibility. Risk factors for Salmonella infection include a variety of foods; for example, contaminated chicken, beef, and pork. Globally, there is a growing incidence and emergence of life-threatening clinical cases, especially due to multidrug-resistant (MDR) Salmonella spp, including strains exhibiting resistance to important antimicrobials such as beta-lactams, fluoroquinolones, and third-generation cephalosporins. In extreme cases, especially in situations involving very difficult-to-treat strains, death usually results. The severity of the infections resulting from Salmonella pathogens is dependent on the serovar type, host susceptibility, the type of bacterial strains, and growth environment. This review therefore aims to detail the nomenclature, etiology, history, pathogenesis, reservoir, clinical manifestations, diagnosis, epidemiology, transmission, risk factors, antimicrobial resistance, public health importance, economic impact, treatment, and control of salmonellosis., Competing Interests: The authors declare that there is no conflict of interest.

The lasso peptide microcin Y (MccY) effectively inhibits various serotypes of Salmonella in vitro, but the antibacterial effect against S. Pullorum in poultry is still unclear. This study was the first to evaluate the safety and anti-S. Pullorum infection of MccY in specific pathogen-free (SPF) chicks. The safety test showed that the body weight, IgA and IgM levels of serum, and cecal microbiota structure of 3 groups of chicks orally administrated with different doses of MccY (5 mg/kg, 10 mg/kg, 20 mg/kg) for 14 days were not significantly different from those of the control group. Then, the chicks were randomized into 3 groups for the experiment of anti-S. Pullorum infection: (I) negative control group (NC), (II) S. Pullorum-challenged group (SP, 5 × 10 8  CFU/bird), (III) MccY-treated group (MccY, 20 mg/kg). The results indicated that compared to the SP group, treatment of MccY increased body weight and average daily gain (P < 0.05), reduced S. Pullorum burden in feces, liver, and cecum (P < 0.05), enhanced the thymus, and decreased the spleen and liver index (P < 0.05). Additionally, MccY increased the jejunal villus height, lowered the jejunal and ileal crypt depth (P < 0.05), and upregulated the expression of IL-4, IL-10, ZO-1 in the jejunum and ileum, as well as CLDN-1 in the jejunum (P < 0.05) compared to the SP group. Furthermore, MccY increased probiotic flora (Barnesiella, etc.), while decreasing (P < 0.05) the relative abundance of pathogenic flora (Escherichia and Salmonella, etc.) compared to the SP group., (© 2024. The Author(s).)

In the pork production chain, the control at slaughterhouse aims to ensure safe food thanks to proper hygienic conditions during all steps of the slaughtering. Salmonella is one of the main foodborne pathogens in the EU causing a great number of human cases, and pigs also contribute to its spreading. Pig is the main reservoir of the zoonotic hepatitis E virus (HEV) that can be present in liver, bile, feces and even rarely in blood and muscle. The aim of this study was to assess the presence of both Salmonella and HEV in several points of the slaughtering chain, including pig trucks. Other viruses hosted in the gut flora of pigs and shed in feces were also assayed (porcine adenovirus PAdV, rotavirus, norovirus, and mammalian orthoreovirus MRV). Torque teno sus virus (TTSuV) present in both feces, liver and blood was also considered. Four Italian pig abattoirs were sampled in 12 critical points, 5 of which were the outer surface of carcasses before processing. HEV and rotavirus (RVA) were not detected. Norovirus was detected once. Salmonella was detected in two of the 4 abattoirs: in the two lairage pens, in the site of evisceration and on one carcass, indicating the presence of Salmonella if carcass is improper handled. The sampling sites positive for Salmonella were also positive for PAdV. MRV was detected in 10 swabs, from only two abattoirs, mainly in outer surface of carcasses. TTSuV was also detected in all abattoirs. Our study has revealed a diverse group of viruses, each serving as indicator of either fecal (NoV, RVA, PAdV, MRV) or blood contamination (TTSuV). TTSuV could be relevant as blood contamination indicators, crucial for viruses with a viremic stage, such as HEV. The simultaneous presence of PAdV with Salmonella is relevant, suggesting PAdV as a promising indicator for fecal contamination for both bacterial and viruses. In conclusion, even in the absence of HEV, the widespread presence of Salmonella at various points in the chain, underscores the need for vigilant monitoring and mitigation strategies which could be achieved by testing not only bacteria indicators as expected by current regulation, but also some viruses (PAdV, TTSuV, MRV) which could represent other sources of fecal contamination., 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 © 2023. Published by Elsevier Ltd.)

Importance: Salmonella outbreaks linked to poultry meat have been reported continuously worldwide. Therefore, Salmonella contamination of poultry meats in slaughterhouses is one of the critical control points for reducing disease outbreaks in humans., Objective: This study examined the carry-over contamination of Salmonella species through the entire slaughtering process in South Korea., Methods: From 2018 to 2019, 1,097 samples were collected from the nine slaughterhouses distributed nationwide. One hundred and seventeen isolates of Salmonella species were identified using the invA gene-specific polymerase chain reaction, as described previously. The serotype, phylogeny, and antimicrobial resistance of isolates were examined., Results: Among the 117 isolates, 93 were serotyped into Salmonella Mbandaka (n = 36 isolates, 30.8%), Salmonella Thompson (n = 33, 28.2%), and Salmonella Infantis (n = 24, 20.5%). Interestingly, allelic profiling showed that all S . Mbandaka isolates belonged to the lineage of the sequence type (ST) 413, whereas all S . Thompson isolates were ST292. Moreover, almost all S . Thompson isolates (97.0%, 32/33 isolates) belonging to ST292 were multidrug-resistant and possessed the major virulence genes whose products are required for full virulence. Both serotypes were distributed widely throughout the slaughtering process. Pulsed-field gel electrophoretic analysis demonstrated that seven S . Infantis showed 100% identities in their phylogenetic relatedness, indicating that they were sequentially transmitted along the slaughtering processes., Conclusions and Relevance: This study provides more evidence of the carry-over transmission of Salmonella species during the slaughtering processes. ST292 S . Thompson is a potential pathogenic clone of Salmonella species possibly associated with foodborne outbreaks in South Korea., Competing Interests: The authors declare no conflicts of interest., (© 2024 The Korean Society of Veterinary Science.)

Salmonella and Campylobacter are common bacterial hazards causing foodborne illnesses worldwide. A large proportion of Salmonella and Campylobacter illnesses are attributed to contaminated poultry products that are mishandled or under cooked. Processing interventions such as chilling and post-chill dip are critical to reducing microbial contamination of poultry. A comprehensive search of the literature published between 2000 and 2021 was conducted in the databases Web of Science, Academic Search Complete, and Academic OneFile. Studies were included if they were in English and investigated the effects of interventions against Salmonella and/or Campylobacter on whole carcasses and/or parts during the chilling or post-chill stages of poultry processing. Random-effects meta-analyses were performed using the "meta" package in the R programming language. Subgroup analyses were assessed according to outcome measure reported, microorganism tested, processing stage assessed, and chemical treatment used. The results included 41 eligible studies. Eighteen studies reported results of 28 separate interventions against Salmonella and 31 reported results of 50 separate interventions against Campylobacter. No significant difference (P> 0.05) was observed when comparing the combined mean difference of all interventions targeting Salmonella to the combined mean difference of all interventions targeting Campylobacter or when comparing chilling times within each pathogen subgroup. For analyses examining antimicrobial additives, peroxyacetic acid (PAA) had the largest reduction against Salmonella population regardless of chilling time (P< 0.05). PAA also had the largest reduction against Campylobacter population and prevalence during primary chilling (P< 0.01). Air chilling showed a lower reduction for Campylobacter than any immersion chilling intervention (P< 0.05). Chilling time and antimicrobial used during poultry processing had varying effects depending on the pathogen and outcome measure investigated (concentration or prevalence). High heterogeneity and low sample numbers in most analyses suggest that more high-quality research that is well-designed and has transparent reporting of methodology and results is needed to corroborate the results., Competing Interests: DISCLOSURES The authors declare no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Salmonella is a major food-borne bacterial pathogen that causes food poisoning related to the consumption of eggs, milk, and meat. Food safety in relation to Salmonella is particularly important for eggs because their shells as well as their contents can be a source of contamination. Chicken can also be infected with influenza virus, but it remains unclear how co-infection of Salmonella and influenza virus affect each other., Aim: The potential influence of co-infection of Salmonella and influenza virus was examined., Methods: Salmonella Abony and influenza virus were injected into chicken embryonated eggs. After incubation, proliferation of Salmonella and influenza virus was measured using a direct culture assay for bacteria and an enzyme-linked immunosorbent assay for influenza virus, respectively., Results: Our findings indicate that the number of colony-forming units (CFUs) of Salmonella did not vary between chicken embryonated eggs co-infected with influenza A virus and Salmonella -only infected eggs. Furthermore, we found the proliferation of influenza A or B virus was not significantly influenced by co-infection of the eggs with Salmonella ., Conclusion: These results suggest that combined infection of Salmonella with influenza virus does not affect each other, at least in terms of their proliferation., Competing Interests: The Author declares that there is no conflict of interest.

Salmonella enterica is a prominent cause of foodborne disease in the United States. However, the mechanism and route of pathogen transmission that leads to Salmonella infection in commercial processing plants are poorly understood. This study aimed to investigate the effect of mixed-species biofilms on S. enterica survival and persistence under sanitizer stress [Quaternary ammonium compounds (QACs)] by analyzing 78 floor drain samples from a meat processing facility and three S . enterica strains (serovars Cerro, Montevideo, and Typhimurium) isolated from that facility and an unrelated source. The four test groups were as follows: control, QAC treatment, Salmonella addition, and QAC treatment with Salmonella addition. DNAs were extracted, and 16S rRNA gene based on the variable region V4 amplicon sequencing was performed to analyze the relative abundance, core microbiome, and Alpha and Beta diversity using the qiime2 pipeline. At the genus level, the Brochothrix (45.56%), Pseudomonas (38.94%), Carnobacterium (6.18%), Lactococcus (4.68%), Serratia (3.14%), and Staphylococcus (0.82%) were shown to be the most prevalent in all drain samples. The results demonstrate that the relative abundance of different bacterial genera was affected by both QAC treatment and Salmonella addition, with some genera showing increases or decreases in abundance. Notably, the correlation network was constructed to understand the relationships between the different bacteria. Nitrospira had the greatest number of connections in the floor drain environment network, with two negative and eight positive correlations. The results suggest that Nitrospira in the mixed-species biofilm community may play a role in converting ammonium in the QAC sanitizer into nitrites. Thus, Nitrospira could be a potentially important genus in providing sanitizer resistance to pathogen-encompassed mixed-species biofilms.IMPORTANCE Salmonella contamination in meat processing facilities can lead to foodborne illness outbreaks. Our study characterized the microbiome dynamics in beef facility drains and their response to Salmonella addition and common sanitizer (QAC). Nitrospira could be an important genus in providing sanitizer resistance to pathogen-encompassed mixed-species biofilms. The results provide insight into the impact of mixed-species biofilms on Salmonella survival and persistence under sanitizer stress in meat processing facilities. The results highlight the need to consider mixed-species biofilm effects when developing targeted interventions to enhance food safety., Competing Interests: The authors declare no conflict of interest.

Quantitative microbial risk assessment (QMRA) has witnessed rapid development within the context of food safety in recent years. As a means of contributing to these advancements, a QMRA for Salmonella spp. in fresh chicken patties for the general European Union (EU) population was developed. A two-dimensional (Second Order) Monte-Carlo simulation method was used for separating variability and uncertainty of model's parameters. The stages of industrial processing, retail storage, domestic storage, and cooking in the domestic environment were considered in the exposure assessment. For hazard characterization, a dose-response model was developed by combining 8 published dose-response models using a Pert distribution for describing uncertainty. The QMRA model predicted a mean probability of illness of 1.19*10 -4 (5.28*10 -5 - 3.57*10 -4 95 % C.I.), and a mean annual number of illnesses per 100,000 people of 2.13 (0.96 - 6.59 95 % C.I.). Moreover, sensitivity analysis was performed, and variability in cooking preferences was found to be the most influential model parameter (r = -0.39), followed by dose-response related variability (r = 0.22), and variability in the concentration of Salmonella spp. at the time of introduction at the processing facility (r = 0.11). Various mitigation strategy scenarios were tested, from which, "increasing the internal temperature of cooking" and "decreasing shelf life" were estimated to be the most effective in reducing the predicted risk of illness. Salmonella-related illnesses exhibit particularly high severity, making them some of the most prominent zoonotic diseases in the EU. Regular monitoring of this hazard in order to further highlight its related parameters and causes is a necessary procedure. This study not only provides an updated assessment of Salmonella spp. risk associated with chicken patties, but also facilitates the identification of crucial targets for scientific investigation and implementation of real-world intervention strategies., 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.)

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.

The trend of feeding dogs and cats with raw pet food claiming health benefits poses health concerns due to the occurrence of pathogenic bacteria. High pressure processing (HPP) allows the non-thermal inactivation of microorganisms, preserving the nutritional characteristics with minimal impact on organoleptic traits of food. The present study aimed to evaluate and model the effect of HPP application (450-750 MPa for 0-7 min) on the inactivation of Salmonella, endogenous microbiota and colour of raw pet food formulated with different concentrations of lactic acid (0-7.2 g/kg) as natural antimicrobial. Additionally, the effect of a subsequent frozen storage of pressurized product was assessed. Salmonella inactivation ranged between 1 and 9 log, depending on the combination of conditions. According to the polynomial model obtained, the effect of pressure was linear, while a quadratic term was also included for holding time (depicting the occurrence of a resistant tail at ca. 4-6 min). The effect of lactic acid was dependent on the pressure level, being most relevant for treatments below 600 MPa. Frozen storage after HPP prevented the pathogen recovery and caused a further Salmonella inactivation enhanced by lactic acid in most of the treatments. Endogenous microbial groups were significantly reduced by HPP to below the detection level in several conditions. In general, little effect of HPP on the instrumental colour parameters was observed, except for a slight increase in lightness, which was hardly appreciable from visual observation. High pressure processing emerges as a relevant technology for the control of Salmonella spp. and to manage the microbiological safety of raw pet food. The mathematical model can be used as decision support tool to design safer raw pet food, while keeping the desired freshness appearance of the products., Competing Interests: Declaration of competing interest Authors declare no conflict of interest. The funders provided the raw materials for preparing the raw pet food product used in the study. They had no responsibility on the design of experiments, data collection and analysis or decision to publish., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Abstract: This multi-institutional study assessed the efficacy of Enterococcus faecium NRRL B-2354 as a nonpathogenic Salmonella surrogate for thermal processing of nonfat dry milk powder, peanut butter, almond meal, wheat flour, ground black pepper, and date paste. Each product was analyzed by two laboratories (five independent laboratories total), with the lead laboratory inoculating (E. faecium or a five-strain Salmonella enterica serovar cocktail of Agona, Reading, Tennessee, Mbandaka, and Montevideo) and equilibrating the product to the target water activity before shipping. Both laboratories subjected samples to three isothermal treatments (between 65 and 100°C). A log-linear and Bigelow model was fit to survivor data via one-step regression. On the basis of D80°C values estimated from the combined model, E. faecium was more thermally resistant (P < 0.05) than Salmonella in nonfat dry milk powder (DEf-80°C, 100.2 ± 5.8 min; DSal-80°C, 28.9 ± 1.0 min), peanut butter (DEf-80°C, 133.5 ± 3.1 min; DSal-80°C, 57.6 ± 1.5 min), almond meal (DEf-80°C, 34.2 ± 0.4 min; DSal-80°C, 26.1 ± 0.2 min), ground black pepper (DEf-80°C, 3.2 ± 0.8 min; DSal-80°C, 1.5 ± 0.1 min), and date paste (DEf-80°C, 1.5 ± 0.0 min; DSal-80°C, 0.5 ± 0.0 min). Although the combined laboratory D80°C for E. faecium was lower (P < 0.05) than for Salmonella in wheat flour (DEf-80°C, 9.4 ± 0.1 min; DSal-80°C, 10.1 ± 0.2 min), the difference was ∼7%. The zT values for Salmonella in all products and for E. faecium in milk powder, almond meal, and date paste were not different (P > 0.05) between laboratories. Therefore, this study demonstrated the impact of standardized methodologies on repeatability of microbial inactivation results. Overall, E. faecium NRRL B-2354 was more thermally resistant than Salmonella, which provides support for utilizing E. faecium as a surrogate for validating thermal processing of multiple low-moisture products. However, product composition should always be considered before making that decision., (Published 2022 by the International Association for Food Protection. Not subject to U.S. Copyright.)

To counteract the dramatic increase in antibiotic-resistant bacterial pathogens, many countries, including China, have banned the use of antibiotic-supplemented feed for farming animals. However, the exact consequences of this policy have not been systematically evaluated. Therefore, Salmonella isolates from farms that ceased using antibiotics 1-5 years ago were compared with isolates from farms that continue to use antimicrobials as growth promotors. Here, we used whole-genome sequencing combined with in-depth phenotypic assays to investigate the ecology, epidemiology, and persistence of multi-drug resistant (MDR) Salmonella from animal farms during the withdrawal of antibiotic growth promotors. Our results showed that the prevalence of Salmonella was significantly lower in antibiotic-free feed (AFF) farms compared to conventional-feed (CF) farms, even though all isolates obtained from AFF farms were MDR (>5 classes) and belonged to well-recognized predominant serovars. The additional phylogenomic analysis combined with principal component analysis showed high similarity between the predominant serovars in AFF and CF farms. This result raised questions regarding the environmental persistence capabilities of MDR strain despite AFF policy. To address this question, a representative panel of 20 isolates was subjected to disadvantageous environmental stress assays. These results showed that the predominant serovars in AFF and CF farms were more tolerant to stress conditions than other serovars. Collectively, our findings suggest that AFF helps eliminate only specific MDR serovars, and future guiding policies would benefit by identifying predominant Salmonella clones in problematic farms to determine the use of AFF and additional targeted interventions., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Fine ground black pepper generally consumed as a seasoning without any further processing has been associated with Salmonella enterica outbreaks. Thermal inactivation kinetics data is necessary to develop a pasteurization process for fine ground black pepper. This study investigates the influence of temperature and water activity on thermal inactivation kinetics of Salmonella in fine ground black pepper. It also assesses the suitability of Enterococcus faecium as a surrogate for Salmonella. Fine ground black pepper of varying water activities, a w (0.40, 0.55, 0.70) was subjected to isothermal treatments at different temperatures (65-80 °C) for five equidistant time points with intervals ranging from 18 s to 250 min. The survival data were used to fit two primary models (log-linear and Weibull) and two secondary models (response surface and Modified Bigelow). Results indicated that among the two primary models, the Weibull model explained the thermal inactivation kinetics better with lower RMSE (0.24 - 0.56 log CFU/g) and AIC c values at all a w and temperatures. Water activity and treatment temperature significantly enhanced the thermal inactivation of Salmonella. E. faecium NRRL B-2354 was found to be a suitable surrogate for Salmonella in fine ground black pepper at all tested treatment conditions. The developed modified Bigelow model based on the Weibull model could be applied to predict the inactivation kinetics of Salmonella in black pepper and would benefit the spice industry in identifying process parameters for thermal pasteurization of fine ground black pepper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Thermal inactivation of pathogenic and spoilage organisms in low and intermediate moisture foods is of critical importance for guaranteeing microbiological safety and stability of these products. Producers tendentially reduce salt in low and intermediate moisture foods because of nutritional health considerations, but it is unclear how this affects microbial inactivation rates during pasteurization. In this study we predict the time to achieve a pre-defined 6-log reduction for Salmonella enterica subsp. enterica serovar Napoli (hereafter: S. Napoli) and Eurotium herbariorum mould spores (hereafter: E. herbariorum spores) and the relationship with product characteristics. We tested 31 design products for heat inactivation of S. Napoli and 29 design products for heat inactivation of E. herbariorum spores. We used a global Bayesian regression combining primary Weibull models with a secondary regression model to relate pasteurization temperature and product characteristics (water activity (a w ), pH, and fractions of sodium chloride, sucrose and oil on product) to microbial counts. With this model, we predict the time to 6-log reduction. Thermal inactivation rates were much higher for vegetative S. Napoli than for E. herbariorum spores. Also, inactivation curves were non-linear for many experiments. There were significant associations between the Weibull model parameters and temperature, and pH and a w for S. Napoli and E. herbariorum spores, respectively. We parameterized an inactivation model for S. Napoli and E. herbariorum spores using design products with a broad range of characteristics and showed how the simplified approach of using D-values does not accurately describe the non-linearity of thermal inactivation for both types of organism. Results of our model can be used to produce accurate heat inactivation predictions as input for the pasteurization process in factories where intermediate moisture foods are manufactured., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Adequate surrogate identification is critical for validating in-plant thermal process controls for Salmonella inactivation in different food matrices. This study compared the thermal inactivation parameters (D- and z-values) and evaluated the heat resistance of Enterococcus faecium (8459) as a surrogate for a 5-serovar Salmonella cocktail in cornmeal. The cornmeal was spray inoculated with the respective bacteria to achieve ~9 log CFU/g population and set to the desired moisture contents (16, 22, and 28% w.b.). The inoculated cornmeal was then heat-treated at pre-determined temperatures (60, 64, and 68 °C) in sealed aluminum thermal-death-time disks in hot water baths for pre-determined time intervals. Injury-recovery media [brain heart infusion (BHI) agar overlaid with xylose lysine deoxycholate (XLD) agar for Salmonella or BHI agar overlaid m-enterococcus agar for E. faecium] were used for microbial enumeration to account for thermally injured bacterial cells. The D-values of Salmonella in cornmeal at 16, 22, and 28% moisture content were 37.5, 8.4, and 2.4 min at 60 °C, 19.9, 3.5, and 1.1 min at 64 °C, and 10.1, 1.4, and 0.5 min at 68 °C, respectively. The D-values of E. faecium in cornmeal at 16, 22, and 28% moisture content were 140.4, 18.9, and 3.3 min at 60 °C, 78.4, 7.1, and 1.6 min at 64 °C, and 37.3, 2.8, and 0.8 min at 68 °C, respectively. The z-values of E. faecium and Salmonella in cornmeal at 16, 22, and 28% moisture content were 13.9, 9.7, and 12.5 °C, and 14.0, 10.4, and 11.7 °C, respectively. These results indicated similar or higher thermal resistance (D-values) and equivalent thermal sensitivity (z-values) of E. faecium compared to Salmonella at different moisture contents and respective temperatures (P ≤ 0.05). Therefore, E. faecium could be used as a surrogate for Salmonella during thermal process validation of cornmeal processing., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Abstract: Meat contamination by Salmonella enterica is a serious public health concern. Available data have suggested that biofilm formation at processing plants and contaminated contact surfaces might contribute to meat contamination. Because transfer from contact surfaces to food products via direct contact has been deemed the most common bacteria transmission route that can lead to contamination, we evaluated the effect of Salmonella biofilm forming ability, contact surface material, and beef surface tissue type on Salmonella biofilm transfer from hard surfaces to beef products. Salmonella biofilms developed on the common contact surfaces stainless steel (SS) and polyvinylchloride (PVC) were transferred consecutively via 30 s of direct contact to either lean muscle or adipose tissue surfaces of 15 pieces of beef trim. The Salmonella biofilm cells could be effectively transferred multiple times from the contact surfaces to the beef trim as indicated by quantifiable Salmonella cells on most meat samples. Biofilm forming ability had the most significant impact (P < 0.05) on transfer efficiency. More cells of Salmonella strains that formed strong biofilms were transferred after each contact and contaminated more meat samples with quantifiable cells compared with strains that formed weak biofilms. Contact surface materials also affected transferability. Salmonella biofilms on SS transferred more efficiently than did those on PVC. In contrast, the two types of meat surface tissues were not significantly different (P > 0.05) in biofilm transfer efficiency. Beef trim samples that were in contact with biofilms but did not have quantifiable Salmonella cells were positive for Salmonella after enrichment culture. Our results indicate the high potential of Salmonella biofilms on common contact surfaces in meat processing plants to cause product cross-contamination., (Published 2022 by the International Association for Food Protection. Not subject to U.S. Copyright.)

Salmonella can become viable but nonculturable (VBNC) in response to environmental stressors, but the induction of the VBNC state in Salmonella contaminating ready-to-eat dried fruit is poorly characterized. Dried apples, strawberries, and raisins were mixed with a five-strain cocktail of Salmonella at 4% volume per weight of dried fruit at 10 9 CFU/g. The inoculated dried fruit were then dried in desiccators at 25°C until the water activity ( a w ) approximated that of the uninoculated dried fruit. However, Salmonella could not be recovered after drying, not even after enrichment, suggesting a population reduction of approximately 8 log CFU/g. To assess the potential impact of storage temperature on survival, dried apples were spot-inoculated with the Salmonella cocktail, dried under ambient atmosphere at 25°C, and stored at 4 and 25°C. Spot inoculation permitted recovery of Salmonella on dried apple after drying, with the population of Salmonella decreasing progressively on dried apples stored at 25°C until it was undetectable after about 46 days, even following enrichment. The population decline was noticeably slower at 4°C, with Salmonella being detected until 82 days. However, fluorescence microscopy and laser scanning confocal microscopy with the LIVE/DEAD Bac Light bacterial viability system at time points at which no Salmonella could be recovered on growth media even following enrichment showed that a large proportion (56 to 85%) of the Salmonella cells on the dried fruit were viable. The data suggest that the unique combination of stressors in dried fruit can induce large numbers of VBNC cells of Salmonella. IMPORTANCE Salmonella is a leading foodborne pathogen globally causing numerous outbreaks of foodborne illnesses and remains the leading contributor to deaths attributed to foodborne disease in the United States and other industrialized nations. Therefore, efficient detection methods for Salmonella contaminating food are critical for public health and food safety. Culture-based microbiological methods are considered the gold standard for the detection and enumeration of Salmonella in food. Findings from this study suggest that unique stressors on dried fruit can induce the VBNC state in Salmonella, thus rendering it undetectable with culture-based methods even though the bacteria remain viable. Therefore, strong consideration should be given to using, in addition to culture-based methods, microscopic and molecular methods for the accurate detection of all viable and/or culturable cells of Salmonella contaminating dried fruit, as all of these cells have the potential to cause human illness.

Salmonella invades and disrupts gut epithelium integrity, creating an infection-generated electric field that can drive directional migration of macrophages, a process called galvanotaxis. Phagocytosis of bacteria reverses the direction of macrophage galvanotaxis, implicating a bioelectrical mechanism to initiate life-threatening disseminations. The force that drives direction reversal of macrophage galvanotaxis is not understood. One hypothesis is that Salmonella can alter the electrical properties of the macrophages by modifying host cell surface glycan composition, which is supported by the fact that cleavage of surface-exposed sialic acids with a bacterial neuraminidase severely impairs macrophage galvanotaxis, as well as phagocytosis. Here, we utilize N-glycan profiling by nanoLC-chip QTOF mass cytometry to characterize the bacterial neuraminidase-associated compositional shift of the macrophage glycocalyx, which revealed a decrease in sialylated and an increase in fucosylated and high mannose structures. The Salmonella nanH gene, encoding a putative neuraminidase, is required for invasion and internalization in a human colonic epithelial cell infection model. To determine whether NanH is required for the Salmonella infection-dependent direction reversal, we constructed and characterized a nanH deletion mutant and found that NanH is partially required for Salmonella infection in primary murine macrophages. However, compared to wild type Salmonella, infection with the nanH mutant only marginally reduced the cathode-oriented macrophage galvonotaxis, without canceling direction reversal. Together, these findings strongly suggest that while neuraminidase-mediated N-glycan modification impaired both macrophage phagocytosis and galvanotaxis, yet to be defined mechanisms other than NanH may play a more important role in bioelectrical control of macrophage trafficking, which potentially triggers dissemination.

Critical cancer pathways often cannot be targeted because of limited efficiency crossing cell membranes. Here we report the development of a Salmonella-based intracellular delivery system to address this challenge. We engineer genetic circuits that (1) activate the regulator flhDC to drive invasion and (2) induce lysis to release proteins into tumor cells. Released protein drugs diffuse from Salmonella containing vacuoles into the cellular cytoplasm where they interact with their therapeutic targets. Control of invasion with flhDC increases delivery over 500 times. The autonomous triggering of lysis after invasion makes the platform self-limiting and prevents drug release in healthy organs. Bacterial delivery of constitutively active caspase-3 blocks the growth of hepatocellular carcinoma and lung metastases, and increases survival in mice. This success in targeted killing of cancer cells provides critical evidence that this approach will be applicable to a wide range of protein drugs for the treatment of solid tumors., (© 2021. The Author(s).)

In light of emerging antibiotic resistance, synthesis of active, environmental friendly antimicrobial alternatives becomes increasingly necessary. In this study, ZnO quantum dots (ZnO QDs) were developed by the sol-gel method and characterized. The antibacterial activities of ZnO QDs against Escherichia coli ( E. coli ), Staphylococcus aureus ( S. aureus ) and Salmonella Pullorum ( S. Pullorum ) were systematically investigated. Moreover, the protective effects of ZnO QDs on Salmonella-caused pullorosis in chicks were also explored. The results indicated that the size range of ZnO QDs was 3-6 nm. Antibacterial results showed that ZnO QDs treatment inhibited the growth of E. coli , S. aureus , and S. Pullorum in the rate of 87.06 ± 0.98%, 94.75 ± 2.28%, and 85.55 ± 1.15%, respectively. Its excellent antibacterial property was manifested with the minimum inhibitory concentration of 0.7812, 0.0976, and 0.1953 mg ml -1 , which may be attributed to the production of reactive oxygen species, the dissolution of Zn 2+ ions, and the loss of cell integrity. Furthermore, in the in vivo test, the ZnO QDs effectively reduced the mortality of chicks infected with S. Pullorum via regulating the balance of the intestinal flora, protecting liver and intestine, and modulating the balance of antioxidation systems. This study reveals that ZnO QDs exerts remarkably antibacterial activity in vitro and anti-pullorosis effect in chicks., (© 2021 IOP Publishing Ltd.)

Efficient in vivo delivery of a CRISPR/Cas9 plasmid is of paramount importance for effective therapy. Here, we investigated the usability of Salmonella as a plasmid carrier for in vivo therapy against virus-induced cancer using Marek's disease virus (MDV) as a model for study in chickens. A green fluorescent protein-expressing CRISPR/Cas9 plasmid encoding the virulence gene pp38 was constructed against Marek's disease virus. Therapeutic plasmids were transformed into Salmonella carrying lon and sifA gene deletions. The animals in 5 groups were intraperitoneally inoculated with phosphate-buffered saline, vector control, or Salmonella before or after MDV infection, or left uninfected as a naïve control. Therapeutic effectiveness was evaluated by observing disease outcomes and the viral copy number in peripheral blood mononuclear cells. The efficacy of plasmid delivery by Salmonella was 13 ± 1.7% in the spleen and 8.0 ± 1.8% in the liver on the 6 th day post-infection. The Salmonella-treated groups showed significant resistance to MDV infection. The maximum effect was observed in the group treated with Salmonella before MDV infection. None of the chickens fully recovered; however, the results suggested that timely delivery of Salmonella could be effective for in vivo CRISPR/Cas9-mediated genetic interference against highly pathogenic MDV. The use of Salmonella in CRISPR systems provides a simpler and more efficient platform for in vivo therapy with CRISPR than the use of conventional in vivo gene delivery methods and warrants further development., (© 2021. The Author(s).)

Salmonella enterica ssp. enterica serovars Enteritidis (SE) and Gallinarum (SG) cause different diseases in chickens. However, both are able to reach the blood stream where heterophils and monocytes are potentially able to phagocytose and kill the pathogens. Using an ex vivo chicken whole blood infection model, we compared the complex interactions of the differentially host-adapted SE and SG with immune cells in blood samples of two White Leghorn chicken lines showing different laying performance (WLA: high producer; R11: low producer). In order to examine the dynamic interaction between peripheral blood leucocytes and the Salmonella serovars, we performed flow cytometric analyses and survival assays measuring (i) leucocyte numbers, (ii) pathogen association with immune cells, (iii) Salmonella viability and (iv) immune gene transcription in infected whole blood over a four-hour co-culture period. Inoculation of blood from the two chicken lines with Salmonella led primarily to an interaction of the bacteria with monocytes, followed by heterophils and thrombocytes. We found higher proportions of monocytes associated with SE than with SG. In blood samples of high producing chickens, a decrease in the numbers of both heterophils and Salmonella was observed. The Salmonella challenge induced transcription of interleukin-8 (IL-8) which was more pronounced in SG- than SE-inoculated blood of R11. In conclusion, the stronger interaction of monocytes with SE than SG and the better survivability of Salmonella in blood of low-producer chickens shows that the host-pathogen interaction and the strength of the immune defence depend on both the Salmonella serovar and the chicken line., (© 2021. The Author(s).)

Nontyphoidal Salmonella (NTS) is one of the major causes of bacterial foodborne infection. It is mainly manifested by self-limiting gastroenteritis in healthy individuals but can also cause severe complications including blood stream infection and mortality. The emergence of multidrug-resistant strains of Salmonella is becoming a global public health concern. This study is aimed at estimating the prevalence of Salmonella , identifying serotypes involved, and investigating antimicrobial susceptibility of the isolates among diarrheic patients attending private hospitals in Addis Ababa. We collected a total of 298 stool samples from diarrheic patients attending five private hospitals in Addis Ababa and isolated Salmonella according to standard microbiological techniques; the isolates were serotyped using slide agglutination and microplate agglutination techniques. Antimicrobial susceptibility test of the isolates was carried out using Kirby-Bauer disc diffusion assay according to Clinical Laboratory Standards Institute guidelines. Fourteen stool samples (4.7%) were positive for Salmonella , and Salmonella Kiambu was the most dominant serovar ( n = 7, 50%) followed by S . Saintpaul ( n = 4, 28.6%) and S. Haifa ( n = 2, 14.3%). Three (21.4%) of the isolates were resistant to sulfisoxazole and tetracycline each and 2 (14.3%) to ampicillin. Resistance to two antimicrobials was detected only in 2 (14.3%) of the isolates, and none of the isolates were resistant to more than two antimicrobials. In conclusion, the current study showed low prevalence of NTS in diarrheic patients attending private hospitals in Addis Ababa. Although multidrug resistance to several antimicrobials was not detected in the isolates, prudent use of antimicrobials is recommended to guaranty the long-term use of the available antimicrobials., Competing Interests: The authors declare that they have no competing interests., (Copyright © 2021 Ruhama Kebede et al.)

Variation observed in heat inactivation of Salmonella strains (data from Combase) was characterized using multilevel modeling with two case studies. One study concerned repetitions at one temperature, the other concerned isothermal experiments at various temperatures. Multilevel models characterize variation at various levels and handle dependencies in the data. The Weibull model was applied using Bayesian regression. The research question was how parameters varied with experimental conditions and how data can best be analyzed: no pooling (each experiment analyzed separately), complete pooling (all data analyzed together) or partial pooling (connecting the experiments while allowing for variation between experiments). In the first case study, level 1 consisted of the measurements, level 2 of the group of repetitions. While variation in the initial number parameter was low (set by the researchers), the Weibull shape factor varied for each repetition from 0.58-1.44, and the rate parameter from 0.006-0.074 h. With partial pooling variation was much less, with complete pooling variation was strongly underestimated. In the second case study, level 1 consisted of the measurements, level 2 of the group of repetitions per temperature experiment, level 3 of the cluster of various temperature experiments. The research question was how temperature affected the Weibull parameters. Variation in initial numbers was low (set by the researchers), the rate parameter was obviously affected by temperature, the estimate of the shape parameter depended on how the data were analyzed. With partial pooling, and one-step global modeling with a Bigelow-type model for the rate parameter, shape parameter variation was minimal. Model comparison based on prediction capacity of the various models was explored. The probability distribution of calculated decimal reduction times was much narrower using multilevel global modeling compared to the usual single level two-step approach. Multilevel modeling of microbial heat inactivation appears to be a suitable and powerful method to characterize and quantify variation at various levels. It handles possible dependencies in the data, and yields unbiased parameter estimates. The answer on the question "to pool or not to pool" depends on the goal of modeling, but if the goal is prediction, then partial pooling using multilevel modeling is the answer, provided that the experimental data allow that., (Copyright © 2021 The Author. Published by 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.)

Salmonella enterica serovar Gallinarum is a host-restricted bacterial pathogen that causes a serious systemic disease exclusively in birds of all ages. Salmonella enterica serovar Typhimurium is a host-generalist serovar. Dendritic cells (DCs) are key antigen-presenting cells that play an important part in Salmonella host-restriction. We evaluated the differential response of chicken blood monocyte-derived dendritic cells (chMoDCs) exposed to S. Gallinarum or S. Typhimurium. S. Typhimurium was found to be more invasive while S. Gallinarum was more cytotoxic at the early phase of infection and later showed higher resistance against chMoDCs killing. S. Typhimurium promoted relatively higher upregulation of costimulatory and other immune function genes on chMoDCs in comparison to S. Gallinarum during early phase of infection (6 h) as analyzed by real-time PCR. Both Salmonella serovars strongly upregulated the proinflammatory transcripts, however, quantum was relatively narrower with S. Gallinarum. S. Typhimurium-infected chMoDCs promoted relatively higher proliferation of naïve T-cells in comparison to S. Gallinarum as assessed by mixed lymphocyte reaction. Our findings indicated that host restriction of S. Gallinarum to chicken is linked with its profound ability to interfere the DCs function. Present findings provide a valuable roadmap for future work aimed at improved vaccine strategies against this pathogen., (© 2021. The Author(s).)

Background: The scientific publications of antimicrobial susceptibilities and resistance must be precise, with interpretations adjusted to the standard. In this frame, knowledge of antimicrobial resistance is fundamental in pathogenic microorganisms such as Salmonella spp., known for many annual deaths worldwide. The objective of this work was to compare the interpretation of standards, the concentrations, and the breakpoints, to study antimicrobial resistance in Non-Typhoidal Salmonella (NTS) isolated from beef, pork, and chicken meat, meat products, and propose additional considerations that improve the use and usefulness of published results., Results: After refining the search based on meeting the inclusion and exclusion criteria, 48 papers were selected. In 33 (68.8%) of them, the disc diffusion method was used, in 11 (22.9%) the MIC determination method, and in 4 (8.33%) were used both. In 24 (50%) of the articles, the selection of a different (correct) standard could have had an impact on the interpretation of antimicrobial susceptibility, which observed when considering three scenarios, i) comparison between the year of the isolation versus the implemented standard, ii) comparison between the year of submission versus implemented standard and iii) comparison between the year of publication versus implemented standard., Conclusions: The most frequent scenario was the inadequate selection of standards, indicating that some studies had not ensured that applied standards kept in line with the date of isolation, date of publication and interpretation of susceptibilities. We proposed 2 years for standards use for resistance and multi-resistance interpretations. On the other hand, we invite researchers to publish their results in the shortest possible time, and editors and reviewers of scientific journals to prioritise these types of studies and verify the correspondence between the standard cited and the one used and the one to be taken into account., (© 2021. The Author(s).)

The effect of ohmic heating (OH) (50, 55, and 60 °C, 6 V/cm) on the inactivation kinetics (Weibull model) and morphological changes (scanning electron microscopy and flow cytometry) of Salmonella spp. in infant formula (IF) was evaluated. In addition, thermal load indicators (hydroxymethylfurfural and whey protein nitrogen index, HMF, and WPNI) and bioactive compounds (DPPH, total phenolics, ACE, α-amylase, and α-glucosidase inhibitory activities) were also studied. OH presented a more intense inactivation rate than conventional heating, resulting in a reduction of about 5 log CFU per mL at 60 °C in only 2.91 min, being also noted a greater cell membrane deformation, higher formation of bioactive compounds, and lower values for the thermal load parameters. Overall, OH contributed to retaining the nutritional value and improve food safety in IF processing., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The effects of the bcsE gene and BcsE protein on bacterial physiology and pathogenicity in SalmonellaTyphimurium and Salmonella Group C1 were investigated. It was observed that biofilm and pellicle formation did not occur in the bcsE gene mutants of wild-type strains. Besides, the 'rdar' (red, dry, rough) biofilm morphotype in wild-type strains changed significantly in the mutants. In terms of the bcsE gene, the swimming and swarming motility in mutant strains showed a dramatic increase compared to the wild-type strains. The Salmonella bcsE gene was cloned into Escherichia coli BL21, and the his-tagged protein produced in this strain was purified to obtain polyclonal antibodies in BALB/c mice. The antibodies were showed labeled antigen specificity to the BscE protein. As a result of immunization and systemic persistence tests carried out with BALB/c mice, BscE protein was determined to trigger high levels of humoral and cellular responses (Th1 cytokine production, IgG2a/IgG1 > 1). Systemic persistence in the liver and spleen samples decreased by 99.99% and 100% in the bcsE mutant strains. Finally, invasion abilities on HT-29 epithelial cells of wild-type strains were utterly disappeared in their bcsE gene mutant strains., (© The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Efforts to overcome resistance to immune checkpoint blockade therapy have focused on vaccination strategies using neoepitopes, although they cannot be applied on a large scale due to the "private" nature of cancer mutations. Here, we show that infection of tumor cells with Salmonella induces the opening of membrane hemichannels and the extracellular release of proteasome-generated peptides by the exacerbation of endoplasmic reticulum (ER) stress. Peptides released by cancer cells foster an antitumor response in vivo, both in mice bearing B16F10 melanomas and in dogs suffering from osteosarcoma. Mass spectrometry analysis on the supernatant of human melanoma cells revealed 12 peptides capable of priming healthy-donor CD8 + T cells that recognize and kill human melanoma cells in vitro and when xenotransplanted in vivo. Hence, we identified a class of shared tumor antigens that are generated in ER-stressed cells, such as tumor cells, that do not induce tolerance and are not presented by healthy cells., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Cooperativity has a central place in biological regulation, providing robust and highly-sensitive regulation. The bacterial flagellar motor implements autonomous torque regulation based on the stator's dynamic structure; the stator units bind to and dissociate from the motor dynamically in response to environmental changes. However, the mechanism of this dynamic assembly is not fully understood. Here, we demonstrate the cooperativity in the stator assembly dynamics. The binding is slow at the stalled state, but externally forced rotation as well as driving by motor torque in either direction boosts the stator binding. Hence, once a stator unit binds, it drives the rotor and triggers the avalanche of succeeding bindings. This cooperative mechanism based on nonequilibrium allostery accords with the recently-proposed gear-type coupling between the rotor and stator.

Salmonella is an intracellular pathogen of a substantial global health concern. In order to identify key players involved in Salmonella infection, we performed a global host phosphoproteome analysis subsequent to bacterial infection. Thereby, we identified the kinase SIK2 as a central component of the host defense machinery upon Salmonella infection. SIK2 depletion favors the escape of bacteria from the Salmonella -containing vacuole (SCV) and impairs Xenophagy, resulting in a hyperproliferative phenotype. Mechanistically, SIK2 associates with actin filaments under basal conditions; however, during bacterial infection, SIK2 is recruited to the SCV together with the elements of the actin polymerization machinery (Arp2/3 complex and Formins). Notably, SIK2 depletion results in a severe pathological cellular actin nucleation and polymerization defect upon Salmonella infection. We propose that SIK2 controls the formation of a protective SCV actin shield shortly after invasion and orchestrates the actin cytoskeleton architecture in its entirety to control an acute Salmonella infection after bacterial invasion., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Salmonellosis in poultry is an important disease that seriously impedes the development of the poultry industry. The increased resistance to antimicrobials against Salmonella has been a major public health concern worldwide. We conducted a study from January to June 2016 in and around the Rajshahi district of Bangladesh on the commercial chicken to isolate, identify and characterize poultry-specific Salmonella, to assess the potential risk factors and to determine the antimicrobial resistance pattern of the isolates. The overall prevalence of Salmonella enterica was 41% (49/120) [95% CI: 31.95%-50.17%] with 41.7% in broiler chicken (25/60) [95% CI: 29.06%-55.12%] and 40% in layer chicken (24/60, 40%) [95% CI: 27.56%-53.46%]. Samples collected from Rajshahi city (OR = 1.37, 95% CI: 0.50-3.73) and Puthia Upazila (OR = 1.51, 95% CI: 0.56-4.12) were more likely to be positive for Salmonella than Charghat Upazila. Salmonella detection was 1.3 times higher in chicken, providing loose feed than those provided ready feed. All the isolates fermented dextrose, maltose and mannitol with the production of acid and gas, but did not ferment sucrose and lactose. The isolates showed catalase, MR, citrate utilization test and TSI agar test positive, but indole and V-P tests negative. Salmonella isolates were sensitive to ciprofloxacin (90%), gentamycin (80%), amoxicillin (75%), streptomycin (70%), ampicillin (45%) and sulfamethoxazole-trimethoprim (45%), whereas highly resistant to penicillin (100%) and nalidixic acid (100%) followed by sulfamethoxazole-trimethoprim (55%), ampicillin (40%) and amoxicillin (25%). Salmonella enterica is endemic in commercial chicken production in Bangladesh with high prevalence. A considerable proportion of Salmonella isolates was found to be resistant to the majority of the common antimicrobial drugs. A good biosecurity system could be effective for the reduction of Salmonella. It is necessary to obtain universal commitments to establish prudent antibiotic use policies., (© 2021 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

The innate and inducible resistance of six Salmonella strains (4/74, FS8, FS115, P167807, ATCC 13076, WT) in mayonnaise at 5 °C following adaptation to different pH/undissociated acetic acid (UAA) combinations (15mM/pH5.0, 35mM/pH5.5, 45mM/pH6.0) was investigated. The inherent and acid-induced responses were strain-dependent. Two strains (ATCC 13076, WT), albeit not the most resistant innately, exhibited the most prominent adaptive potential. Limited/no adaptability was observed regarding the rest strains, though being more resistant inherently. The individual effect of pH and UAA adaptation in the phenotypic and transcriptomic profiles of ATCC 13076 and WT was further examined. The type (pH, UAA) and magnitude of stress intensity affected their responses. Variations in the type and magnitude of stress intensity also determined the relative gene expression of four genes (adiA, cadB, rpoS, ompR) implicated in Salmonella acid resistance mechanisms. adiA and cadB were overexpressed following adaptation to some treatments; rpoS and ompR were downregulated following adaptation to 15mM/pH5.0 and 35mM/pH5.5, respectively. Nonetheless, the transcriptomic profiles did not always correlate with the corresponding phenotypes. In conclusion, strain variations in Salmonella are extensive. The ability of the strains to adapt and induce resistant phenotypes and acid resistance-related genes is affected by the type and magnitude of the stress applied during adaptation., (Copyright © 2020. Published by Elsevier Ltd.)

Thermal inactivation kinetics of Salmonella in low moisture foods are necessary for developing proper thermal processing parameters for pasteurization. The effect of water activity on thermal inactivation kinetics of Salmonella and Enterococcus faecium NRRL B-2354 in ground black pepper has not been studied previously. Identification of a suitable surrogate assists in conducting in-plant process validations. Ground black pepper was inoculated with a 5-serotype Salmonella cocktail or E. faecium NRRL B-2354, equilibrated to water activities of 0.25, 0.45 or 0.65 in a humidity-controlled chamber, and isothermally treated at different temperatures. The survivor data were used for fitting the log-linear models to obtain the D and z-values of Salmonella and E. faecium in ground black pepper. Modified Bigelow models were developed to evaluate the effects of temperature and water activity on the thermal inactivation kinetics of Salmonella and E. faecium. Water activity and temperature showed significant negative effects on the thermal resistance of Salmonella and E. faecium in ground black pepper. For example, significantly higher D values of Salmonella were observed at water activity of 0.45 (D 70°C  = 20.5 min and D 75°C  = 7.8 min) compared to water activity of 0.65 (D 70°C  = 3.9 min and D 75°C  = 2.0 min). D-values of E. faecium were significantly higher than those of Salmonella at all three water activities, indicating that E. faecium is a suitable surrogate for Salmonella in thermal processing validation., (Copyright © 2021. Published by Elsevier B.V.)

Bacterial genotoxins cause DNA damage in eukaryotic cells, resulting in activation of the DNA damage response (DDR) in vitro. These toxins are produced by Gram-negative bacteria, enriched in the microbiota of inflammatory bowel disease (IBD) and colorectal cancer (CRC) patients. However, their role in infection remains poorly characterized. We address the role of typhoid toxin in modulation of the host-microbial interaction in health and disease. Infection with a genotoxigenic Salmonella protects mice from intestinal inflammation. We show that the presence of an active genotoxin promotes DNA fragmentation and senescence in vivo, which is uncoupled from an inflammatory response and unexpectedly associated with induction of an anti-inflammatory environment. The anti-inflammatory response is lost when infection occurs in mice with acute colitis. These data highlight a complex context-dependent crosstalk between bacterial-genotoxin-induced DDR and the host immune response, underlining an unexpected role for bacterial genotoxins., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Salmonella contamination sources and transmission routes were studied in 5 Belgian poultry slaughterhouses. Samples from the slaughter and cutting line after cleaning and disinfection were collected, as well as neck skin samples and thighs during slaughter of the first flock. In total, 680 swab and water samples were taken from the slaughter line before slaughter. In all slaughterhouses, Salmonella was notwithstanding cleaning and disinfection still isolated from the slaughter line before start of activities. The prevalence of Salmonella in the plucking area was 10.4% (38/365) (hanging area: 5.0%, scalding tank: 5.8%, plucking machine: 17.0%); in the evisceration room, 1.5% (2/138); and in the cutting area, 2.0% (3/149). No Salmonella (0/28) was found in samples from the chilling line. On neck skin samples taken from the various lines, Salmonella prevalence was 16.1% (48/299) after plucking, 16.0% (48/300) after evisceration, 23.3% (70/300) after chilling; on thighs, prevalence was 10.0% (24/240). Nine Salmonella serotypes were identified of which Salmonella Infantis was the most common serovar (53.8%), especially in slaughterhouse A. Two contamination causes were identified; first, although all flocks had an official Salmonella negative status, this was in one case incorrect and led to an enormous contamination of the neck skins of the flock and the slaughterline (i.e., cooling water). Second, molecular typing revealed cross-contamination from flocks slaughtered 1 d before sampling. Salmonella was apparently not always eliminated by the cleaning and disinfection process and able to contaminate the carcasses of the first slaughtered flock. In conclusion, the results of this study provided practical insights for poultry production to further improve their Salmonella control, for example, Salmonella status determination closer to the slaughter date, to adapt cleaning and disinfection protocols especially for critical machinery and better hygienic designed equipment., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)