Salmonella Typhimurium Research Articles

STM1863, a Member of the DUFs Protein Family, Is Involved in Environmental Adaptation, Biofilm Formation, and Virulence in Salmonella Typhimurium.

Salmonella Typhimurium (STM) is an important zoonotic Gram-negative pathogen that can cause infection in a variety of livestock and poultry. Meanwhile, as an important foodborne pathogen, the bacterium can survive in various stressful environments and transmits through the fecal-oral route, posing a serious threat to global food safety. To investigate the roles of STM1863, a member of the DUFs protein family, involved in STM environmental adaptation, biofilm formation, and virulence. We analyzed the molecular characteristics of the protein encoded by STM1863 gene and examined intra- and extracellular expression levels of STM1863 gene in mouse macrophages. Furthermore, we constructed STM1863 gene deletion and complementation strains and determined its environmental adaptation under stressful conditions such as acid, alkali, high salt, bile salt, and oxidation. And the capacity of biofilm formation and pathogenicity of those strains were analyzed and compared. In addition, the interaction between the promoter of STM1863 gene and RcsB protein was analyzed using DNA gel electrophoresis migration assay (electrophoretic mobility shift assay [EMSA]). The experiments revealed that acid adaptability and biofilm formation ability of STM1863 gene deletion strain were significantly weakened compared with the parental and complementary strains. Moreover, the adhesion and invasion ability of STM1863 deletion strain to mouse macrophages was significantly decreased, while the median lethal dose (LD50) increased by 2.148-fold compared with the parental strain. In addition, EMSA confirmed that RcsB protein could bind to the promoter sequence of STM1863 gene, suggesting that the expression of STM1863 gene might be modulated by RcsB. The present study demonstrated for the first time that STM1863, a member of the DUFs protein family, is involved in the modulation of environmental adaptation, biofilm formation, and virulence.

Enhancing pathogen identification in cheese with high background microflora using an artificial neural network-enabled paper chromogenic array sensor approach

Biohazards, which may occur at all supply chain stages, pose significant threats to food safety and public health. Addressing these concerns and enhancing food safety necessitates a nondestructive pathogen surveillance approach capable of continuously and simultaneously detecting multiple pathogens. Detecting and differentiating low concentrations of pathogenic bacteria amid high background microflora levels in foods is challenging, requiring technology with high sensitivity and robust discriminatory capability. This study introduces an artificial neural network-driven paper chromogenic array sensor (ANN-PCA) technique developed for the nondestructive, continuous, and simultaneous detection of Salmonella Enteritidis (SE) and Escherichia coli O157:H7 (Ec) from a high background microflora in shredded cheddar cheese. This method enables accurate detection of SE and Ec in monoculture and cocktail culture while distinguishing them from a high level of background microflora (∼7.5 log CFU/g), with accuracies ranging from 72 ± 11% to 92 ± 3%. In addition, SE and Ec were successfully identified at concentrations as low as 1 log CFU/g within one day, with an accuracy of 72 ± 11%. This approach exhibits promising potential for integration into a digitalized, smart, and resilient nondestructive surveillance system for real-time pathogen detection in foods throughout the supply chain without enrichment, incubation, or other sample preparation steps.

Shifts in microbiota and gene expression of nutrient transporters, mucin and interleukins in the gut of fast-growing and slow-growing chickens infected by Salmonella Enteritidis.

Slow-growing breeds are more resistant to Salmonella infection compared to fast-growing broilers. However, it is unclear whether that is associated with innate resistance or rather rely on differences in Salmonella-induced gut responses. We investigated the microbial composition and gene expression of nutrient transporters, mucin, and interleukin in the gut of a fast-growing (Cobb500) and a slow-growing naked neck (NN) chicken breeds challenged with Salmonella Enteritidis. Hatchlings were inoculated at two days of age using sterile broth (sham) or Salmonella Enteritidis (SE) and distributed according to a completely randomized design into four treatments: Cobb-sham; Cobb-SE; NN-sham; and NN-SE. Cecal SE counting and microbial composition by 16S rRNA sequencing were determined at 24-, 96-, and 168-hours post-inoculation (hpi). Gene expression of amino acid (Asct1) and peptide transporters (PepT1), glucose transporters (Sglt1, Glut2 and Glut5) and mucin (Muc2) in the jejunum and expression of interleukins (IL1 beta, IL8, IL17 and IL22) in the cecum was assessed by qPCR at 24 and 168 hpi. NN birds were colonized by SE just as Cobb birds but showed innate upregulation of Muc2, IL8 and IL17 in comparison to Cobb. While nutrient transporter mRNA expression was impaired in SE-challenged Cobb birds, the opposite was observed in NN. There were no differences in microbial diversity at different sampling times for Cobb-SE, whereas the other groups had higher diversity and lower dominance at 24 hpi compared with 96 hpi and 168 hpi. NN birds apparently develop earlier gut microbial stability, have higher basal level of mucin gene expression as well as differential nutrient transporter and interleukin gene expression in the presence of SE which might mitigate the effects of SE infection compared to Cobb birds.

Impacts of on-farm water sanitation practices on microbial hygiene in poultry waterlines and efficacy of sodium hypochlorite-based product on foodborne pathogens

The microbial water quality of poultry water supplies can be correlated to bird health, livability, overall performance, and human food safety. On-farm microbial evaluations were conducted to understand microbial hygiene of poultry waterlines based on water sanitation practices. With on-farm study I, two farms were selected: Farm A and Farm B; that did not practice water sanitation consistently during the flock grow-out period. Prevalence tests for specific pathogens- Salmonella, E coli, Campylobacter, Listeria, and Staphylococcus were performed for both farms by collecting swab samples post bird harvest from waterlines and plating in specific growth media. With the on-farm study II, one broiler farm with four barn units that treated water (Chlorine (Cl)= ∼ 1 ppm) during the flock grow-out period and flushed waterlines between flocks using concentrated chlorine solution (>1000 ppm) was selected. Swab samples (one from each barn, n=4; each sampling occasion) were collected on three occasions-before flushing, after flushing, and at the end of the grow-out period (d 42) to understand biofilm growth nature in poultry waterlines. Additionally, a separate in-vitro study was conducted to understand the efficacy of a commonly used poultry drinking water sanitizer product (sodium hypochlorite, 8.25 %) against specific foodborne pathogens: Salmonella Enteritidis (SE), E. coli and Listeria. Results from the on-farm study I showed that Farm A and Farm B were positive for Listeria and Staphylococcus, whereas these farms were found negative for other species tested. The findings from on-farm study II showed that biofilm reestablished (> 4 log10 cfu/ml) in waterlines by the end of grow-out cycle despite waterline cleaning and consistent water sanitation during flock grow-out period. The in-vitro test showed that the efficacy of chlorine-based water sanitizer at the field application dose rates was affected by pathogen load in water and pathogen types. The overall results from on-farm or in-vitro studies indicated that poultry growers need to emphasize consistent and robust poultry drinking water sanitation practice in their farms to keep the acceptable bacterial levels in poultry water supplies or water systems.

Bioassay guided triterpene isolation and its biological evaluation using branches extract of a significant medicinal plant; Monotheca buxifolia

ObjectiveThe present study focuses on column chromatography of ethyl acetate fractions and re fractions of M.buxifolia branches for potential compound isolation alongside their biological and phytochemical evaluation and TLC analysis. MethodsThe obtained EtOAc fractions were subjected to phytochemical (TPC, TFC), antioxidant (TRP, TAC, DPPH, MC, ABTS), antibacterial and enzyme inhibition (α-amylase, lipase and urease) assays. Fractions indicating the presence of a compound were selected for running successive columns and their factions also analyzed employing the above mentioned bioassays. ResultsA collective none-to-low antibacterial activity was observed. MbBE11 presented the highest antidiabetic potential (79.92%±3.9) besides moderate phytochemical and antioxidant activity. Highest phenolic content (75.85±3.6 μgGAE/mg), flavonoid content (45.39±2μgQE/mg), antioxidant capacity (99.9±3.7 μgAAE/mg), reducing power (94.82±3.9 μgAAE/mg) and radical scavenging activity (DPPH; 86.28±3%, ABTS; 98.9±2.3%, MC; 59.01±2.7%) was reported by MbBE16 making it a potential source of antioxidant agents. Column chromatography of MbBE11 produced two refractions (MbBE21 and MbBE22) which reported moderate phytochemical and high antioxidant activity. High α-amylase and urease inhibition with low lipase inhibition was observed. MbBE21 depicted the largest ZOI against S.enterica (SE) (12±0.7 mm) and MbBE22 against Methicillin-resistant S.aureus (MRSA) (12±0.5 mm). Column chromatography of MbBE16 produced four refractions (MbBE31 – MbBE34) with moderate phytochemical and antioxidant activity without a single dominant fraction. Antibacterial activity remained low with the average ZOI sized at 7±0.5 mm. MbBE15 reported the highest anti-urease at 95.7%±4.7 and significant lipase inhibition at 85.5%±3. MbBE15, MbBE22 MbBE32 resulted in crystal formation later characterized as Lupeol with moderate phytochemical and antioxidant activity. ConclusionThe study concludes that M.buxifolia is a potential source of bioactive components and can be further explored through modern strategies.

A dual RPA-LFD assay for the simultaneous detection of Salmonella typhimurium and Salmonella enteritidis.

Introduction: Salmonella was one of the most common bacteria that caused foodborne illness, with S. typhimurium (Salmonella typhimurium) and S. enteritidis (Salmonella enteritidis) infections accounting for more than 75% of human salmonella infections. Methods: In this study, we developed a method of dual recombinase polymerase amplification (RPA) combined with a lateral flow dipstick for the rapid detection of S. typhimurium and S. enteritidis in clinical specimens (stool). Results: The entire reaction process, including amplification and result reading, could be completed within 65min. The detection limits of S. typhimurium and S. enteritidis in pure culture samples were 5.23 × 101CFU/mL and 3.59 × 101CFU/mL, respectively. The detection limits of S. typhimurium and S. enteritidis in artificially contaminated samples were 8.30 × 101CFU/mL and 2.70 × 102CFU/mL, respectively. In addition, the method had no cross-reaction with other pathogenic microorganisms. The results in clinical samples were fully consistent with those obtained using Bacterial Analysis Manual, with sensitivity and specificity were 100% (8/8) and 100% (17/17) for S. typhimurium and 100% (4/4) and 100% (21/21) for S. enteritidis, respectively. Discussion: The detection limits of S. typhimurium and S. enteritidis in artificially contaminated samples were higher than those in pure culture samples, which might be attributed to the inherent complex composition of artificially contaminated samples. In addition, the detection limits of S. typhimurium and S. enteritidis in the same sample were also different, which might be attributed to different amplification efficiency of two target genes in the same reaction system. Conclusion: This assay had potential application outdoors, as it could be performed within 1h at 38°C without a complex instrument, and the results could be observed with the naked eye. In conclusion, the dual RPA-LFD assay established in this study had practical significance for the rapid detection of S. typhimurium and S. enteritidis in the future.

Antibacterial Effect of Green Synthesis of Selenium Nanoparticles against Salmonella typhi in Small Buffalo

Salmonella typhi is one of the most significant public causes of diarrhea in young animals. Bacterial colonies were first diagnosed by bacteriological methods. The aim of this article is to study the antibacterial effect of green synthesis of selenium nanoparticles against salmonella typhi in small buffalo. From 60 fecal samples of diarrhea, Salmonella typhi consisted of 21(35%) Salmonella paratyphi 9(15%), Escherichia coli 10(16,66%), Shigella 8(13,33%) and Staphylococcus aureus 12(20%). Cinnamon extract was utilized for the green synthesis of selenium nanoparticles (SeNPs). The characterization of green SeNPs was detected using: UV–Visible Spectroscopy. The absorption peak appeared at 298 nm. X-ray diffraction (XRD) indicated that the formed Se-NPs were highly crystalline. FTIR analysis indicated the presence of N-C– and –C-C stretching groups, amines, and Carbohydrates in the composition of SeNPs, by SEM the Se-NPs had a virtually spherical form. The size of the Se-NPs was made from cinnamon extract in the ranges of 34.5 to 45 nm. The antibacterial activity of the cinnamic ethanolic extract and synthesized SeNPs each one alone was investigated against salmonella typhi by using the agar well diffusion method and tested by using a concentration of (25,50,100 μg/mL). The results indicate that Se-NPs gave a good inhibition at 100 mg/ml, which was 14.33 mm, while the cinnamon extract at the same concentration gave an inhibition of 12.33 mm. The broth microdilution method was used to determine the MIC and MBC. MIC values recorded 200 and 25 μg/mL for cinnamon extract and SeNPs Respectively. Additionally, MBC values were recorded at 800 and 200 μg/mL for cinnamon extract and for SeNPs respectively. The RT-PCR was used for the detection of Bss and Bssr gene expression before and after being treated with green synthesized Se-NPs.The results show that the expression of the BssS and BssR gene after treatment with a combination of SeNPs and cinnamon extract increased in the cycle number and gave high gene expression.

Molecular Prediction and Correlation of the Structure and Function of Universal Stress Protein A (UspA) from Salmonella Typhimurium.

Salmonella Typhimurium (ST) is a zoonotic pathogen that can cause gastroenteritis in humans when they consume contaminated food or water. When exposed to various stressors, both from living organisms (biotic) and the environment (abiotic), Salmonella Typhimurium produces Universal Stress Proteins (USPs). These proteins are gaining recognition for their crucial role in bacterial stress resistance and the ability to enter a prolonged state of growth arrest. Additionally, USPs exhibit diverse structures due to the fusion of the USP domain with different catalytic motifs, enabling them to participate in various reactions and cellular activities during stressful conditions. In this particular study, researchers cloned and analyzed the uspA gene obtained from poultry-derived strains of Salmonella Typhimurium. The gene comprises 435 base pairs, encoding a USP family protein consisting of 144 amino acids. Phylogenetic analysis demonstrated a close relationship between the uspA genes of Salmonella Typhimurium and those found in other bacterial species. Weused molecular dynamics simulations and 3D structure prediction to ensure thatthe USPA protein was stable.Furthermore, we alsocarried out motif search and network analysis of protein-protein interactions. The findings from this study offer valuable insights for the development of inhibitors targeted against Salmonella Typhimurium.

Functionalized sampling swabs array-based portable ATP bioluminescence sensor with on-site enrichment and high specificity for live Salmonella detection

Live food-borne pathogens, featured with rapid proliferative capacity and high pathogenicity, pose an emerging food safety and public health crisis. The high-sensitivity detection of pathogens is particularly imperative yet remains challenging. This work developed a functionalized nylon swab array with enhanced affinity for Salmonella typhimurium (S.T.) for high-specificity ATP bioluminescence-based S.T. detection. In brief, the nylon swabs (NyS) were turned to N-methylation nylon (NyS-OH) by reacting with formaldehyde, and NyS-OH were further converted to NyS-CA by reacting with carboxylic groups of citric acid (CA) and EDC/NHS solution, for altering the NyS surface energy to favor biomodification. The antibody-immobilized nylon swab (MNyS-Ab) was ready for S.T.-specific adsorption. Three prepared MNyS-Ab were installed on a stirrer to form an MNyS-Ab array, allowing for on-site enrichment of S.T. through absorptive extraction. The enriched S.T. was quantified by measuring the bioluminescence of ATP released from cell lysis utilizing a portable ATP bioluminescence sensor. The bioassay demonstrated a detectable range of 102–107 CFU mL−1 with a detection limit (LOD) of 8 CFU/mL within 35 min. The signal of single MNyS-Ab swabs was 500 times stronger than the direct detection of 106 CFU/mL S.T. The MNyS-Ab array exhibited a 100-fold increase in extraction level compared to a single MNyS. This combination of a portable bioluminescent sensor and modified nylon swab array offers a novel strategy for point-of-care testing of live S.T. strains. It holds promise for high-sensitivity measurements of other pathogens and viruses.

Effect of a Bacillus subtilis plus Yeast Cell Wall Synbiotic on Salmonella Enteritidis Colonization in Ceca of Layer Pullets

Salmonella Enteritidis (SE) is a major contamination concern in eggs and risk for Salmonellosis in humans. Strains of Bacillus subtilis and yeast cell wall can be used as substitutes for antibiotic substances in feed against Salmonella in poultry. The objective of this study was to assess the effect of BacPack® (Quality Technology International, Inc., Elgin, IL, USA) Q1+1 (BPQ11), a feed additive combination of a Bacillus subtilis strain and Saccharomyces cerevisiae cell wall, on SE cecal colonization in Lohmann LSL pullets. A control group (CON) and a test group (BPQ) were each randomly assigned 100-day-old chicks. CON was fed a corn–soybean meal-based vegetarian mash diet, and BPQ was fed the control diet supplemented with BPQ11 for the duration of the study. At 8 days of age, chicks were orally challenged with a nalidixic acid-resistant SE strain at a dose of 6.3 × 107 colony forming units (CFUs) per bird. At 7, 11, 15, and 19 days post-challenge (DPC), 25 birds per group were euthanized, and their cecal contents were collected and analyzed for SE. SE counts were 6.88, 7.98, 7.79, and 7.50 in CON and 7.18, 7.31, 6.35, and 6.30 log10 CFU/g in BPQ at 7, 11, 15, and 19 DPC, respectively. SE did not differ between CON and BPQ at 7 DPC; however, BPQ had lower (p < 0.0001) SE at 11 (−0.67), 15 (−1.45), and 19 (−1.20 log10 CFU/g) DPC. Results indicate that synbiotic BPQ11 may be a useful dietary pre-harvest tool for SE management in layer birds.

Serum cytokine profile of neonatal broiler chickens infected with Salmonella Typhimurium.

The avian immune system responds to Salmonella infection by expressing cytokines and chemokines. We hypothesized that the immune status of Salmonella Typhimurium (ST) challenged neonatal broilers would differ from the uninfected treatment. The objective of this experiment was to evaluate 12 cytokines. Day of hatch male chicks were randomly allocated into a control or ST challenged group. At day three of age, sterile diluent or 5.0 × 108CFU of ST was given orally to each chick. Blood was obtained 24h post challenge and serum separated for later analysis (n = 30 chicks/treatment). Significant (p ≤ 0.05) increases in pro-inflammatory cytokines-interleukin-6 (IL-6), IL-16, and IL-21; anti-inflammatory cytokines- IL-10; chemokines-regulated on activation, normal T cell expressed and secreted (RANTES), macrophage inflammatory protein-1β (MIP-1β), and MIP-3α; colony stimulating factors-macrophage colony-stimulating factor (M-CSF); and growth factors-vascular endothelial growth factor (VEGF) were observed in the serum of the challenged chicks when compared to the control. No significant differences were observed in IL-2, interferon gamma (IFNγ), and IFNα. These data indicate the detection of mucosal immune responses in broiler chickens following ST infection. The heightened levels of pro-inflammatory cytokines, chemokines, and colony stimulating factors align with known inflammatory mechanisms, like the influx of immune cells. However, the elevation of IL-10 was unexpected, due to its immunoregulatory properties. Notably, the rise in VEGF levels is compelling, as it suggests the possibility of tissue repair and angiogenesis in ST infected birds.

Chitosan-nanoparticle-based oral Salmonella enteritidis subunit vaccine elicits cross-protection against Salmonella typhimurium in broilers

Non-typhoidal Salmonella infection is a significant health and economic burden in poultry industry. Developing an oral vaccine to induce robust mucosal immunity in the intestines of birds, especially cross protection against different Salmonella serotypes is challenging. Therefore, a potent oral vaccine platform that can mitigate different serotypes of Salmonella is warranted for the poultry industry. We reported earlier that the Salmonella enteritidis (SE) immunogenic outer membrane proteins (OMPs) and flagellin (FLA) entrapped in mannose chitosan nanoparticles (OMPs-FLA-mCS NPs) administered prime-boost (d-3 and 3-wk later) by oral inoculation elicits mucosal immunity and reduces challenge SE colonization by over 1 log10 CFU in birds. In this study, we sought to evaluate whether the SE antigens containing OMPs-FLA-mCS NPs vaccine induces cross-protection against Salmonella typhimurium (ST) in broilers. Our data indicated that the OMPs-FLA-mCS NPs vaccine induced higher cross-protective antibody responses compared to commercial Poulvac ST vaccine (contains a modified-live ST bacterium). Particularly, OMPs-FLA-mCS-NP vaccine elicited OMPs and FLA antigens specific increased production of secretory IgA and IgY antibodies in samples collected at both post-vaccination and post-challenge timepoints compared to commercial vaccine group. Notably, the vaccine reduced the challenge ST bacterial load by 0.8 log10 CFU in the cecal content, which was comparable to the outcome of Poulvac ST vaccination. In conclusion, our data suggested that orally administered OMPs-FLA-mCS-NP SE vaccine elicited cross protective mucosal immune responses against ST colonization in broilers. Thus, this candidate vaccine could be a viable option replacing the existing both live and killed Salmonella vaccines for birds.

Multifunctional photoelectrochemical platform based on layer-by-layer assembly of Au/Bi2MoO6/V2CTx for simultaneous detection and inactivation of Salmonella enteritidis

A unique multifunctional photoelectrochemical(PEC) platform for the simultaneous detection and inactivation of Salmonella enteritidis (SE) was proposed. The photoactive material in the platform was a Bi2MoO6/V2CTx composite, and gold nanoparticles were modified to link the aptamer via Au-S bonds. The aptamer was to link the complementary DNA (cDNA) to the electrode surface via base complementarity. In the presence of SE, the aptamer attached to Salmonella enteritidis selectively and then the cDNA was released from the aptamer electrode, the photocurrent signal was amplified, allowing for quantitative detection of Salmonella enteritidis. The linear range of the method was 1.82 × 102 CFU/mL to 1.82 × 108 CFU/mL based on the specific recognition of Salmonella enteritidis by aptamer. The detection limit was 26 CFU/mL. Furthermore, due to its photoelectrocatalytic action, the PEC platform demonstrated strong antibacterial activity (99.99 %). The new technique was created to detect and inactivate Salmonella enteritidis in a timely and sensitive manner.

MARCKS Inhibition Alters Bovine Neutrophil Responses to Salmonella Typhimurium.

Neutrophils are innate immune cells that respond quickly to sites of bacterial infection and play an essential role in host defense. Interestingly, some bacterial pathogens benefit from exuberant neutrophil inflammation. Salmonella is one such pathogen that can utilize the toxic mediators released by neutrophils to colonize the intestine and cause enterocolitis. Because neutrophils can aid gut colonization during Salmonella infection, neutrophils represent a potential host-directed therapeutic target. Myristoylated alanine-rich C-kinase substrate (MARCKS) is an actin-binding protein that plays an essential role in many neutrophil effector responses. We hypothesized that inhibition of MARCKS protein would alter bovine neutrophil responses to Salmonella Typhimurium (STm) ex vivo. We used a MARCKS inhibitor peptide to investigate the role of MARCKS in neutrophil responses to STm. This study demonstrates that MARCKS inhibition attenuated STm-induced neutrophil adhesion and chemotaxis. Interestingly, MARCKS inhibition also enhanced neutrophil phagocytosis and respiratory burst in response to STm. This is the first report describing the role of MARCKS protein in neutrophil antibacterial responses.

STnc1280, a trans-coding sRNA is involved in virulence modulation via targeting gldA mRNA in Salmonella Typhimurium.

Introduction. Salmonella Typhimurium (STM) is a food-borne Gram-negative bacterium, which can infect humans and a wide range of livestock and poultry, causing a variety of diseases such as septicaemia, enteritis and abortion.Hypothesis/Gap Statement. We will decipher the impacts of sRNA STnc1280 on STM virulence and provide a theoretical basis to reveal the regulatory role and molecular mechanism of STnc1280.Aim. The main objective of this study was to clarify whether sRNA STnc1280 exerts regulatory roles on STM pathogenicity.Methodology. The STnc1280 gene was amplified and its molecular characteristics were analysed in this study. Then, STnc1280 gene deletion strain (STM-ΔSTnc1280) and the complementary strain (ΔSTnc1280/STnc1280) were constructed by λ-Red homologous recombination method, respectively, to analyse of adhesion and invasive ability and pathogenicity of different strains. Subsequently, the potential target gene regulated by STnc1280 was predicted using target RNA2 software, followed by the verification of the interaction between STnc1280 and target mRNA using the dual plasmid reporter system (DPRS). Furthermore, the mRNA and protein level of target gene was determined using qRT-PCR and Western blot, respectively.Results. The results revealed that the cell adhesion and invasive ability and pathogenicity of STM-ΔSTnc1280 were significantly reduced compared to STM-SL1344 strain, indicating that the deficiency of STnc1280 gene significantly influenced STM pathogenicity. The DPRS results showed that STnc1280 can interact with the mRNA of target gene gldA, thus suppressing the expression of lacZ gene. Furthermore, the level of gldA mRNA was not influenced in STM-ΔSTnc1280, but the expression of GldA protein decreased significantly.Conclusion. Combining the bioinformatic analysis, these findings suggested that STnc1280 may bind to the SD sequence of gldA mRNA, hindering the binding of ribosomes to gldA mRNA, thereby inhibiting the expression of GldA protein to modulate the virulence of STM.

Establishment and comparison of in situ detection models for foodborne pathogen contamination on mutton based on SWIR-HSI.

Rapid and accurate detection of food-borne pathogens on mutton is of great significance to ensure the safety of mutton and its products and the health of consumers. The feasibility of short-wave infrared hyperspectral imaging (SWIR-HSI) in detecting the contamination status and species of Escherichia coli (EC), Staphylococcus aureus (SA) and Salmonella typhimurium (ST) contaminated on mutton was explored. The hyperspectral images of uncontaminated and contaminated mutton samples with different concentrations (108, 107, 106, 105, 104, 103 and 102 CFU/mL) of EC, SA and ST were acquired. The one dimensional convolutional neural network (1D-CNN) model was constructed and the influence of structure hyperparameters on the model was explored. The effects of different spectral preprocessing methods on partial least squares-discriminant analysis (PLS-DA), support vector machine (SVM) and 1D-CNN models were discussed. In addition, the feasibility of using the characteristic wavelength to establish simplified models was explored. The best full band model was the 1D-CNN model with the convolution kernels number of (64, 16) and the activation function of tanh established by the original spectra, and its accuracy of training set, test set and external validation set were 100.00, 92.86 and 97.62%, respectively. The optimal simplified model was genetic algorithm optimization support vector machine (GA-SVM). For discriminating the pathogen species, the accuracies of SVM models established by full band spectra preprocessed by 2D and all 1D-CNN models with the convolution kernel number of (32, 16) and the activation function of tanh were 100.00%. In addition, the accuracies of all simplified models were 100.00% except for the 1D-CNN models. Considering the complexity of features and model calculation, the 1D-CNN models established by original spectra were the optimal models for pathogenic bacteria contamination status and species. The simplified models provide basis for developing multispectral detection instruments. The results proved that SWIR-HSI combined with machine learning and deep learning could accurately detect the foodborne pathogen contamination on mutton, and the performance of deep learning models were better than that of machine learning. This study can promote the application of HSI technology in the detection of foodborne pathogens on meat.

Fabrication of CMC/PVA/Dextrin-Based Polymeric Membrane for Controlled Release of Cefixime With Enhanced Antibacterial Activity.

This study focuses on the investigation of the significance of polymers in drug delivery approaches. The carboxymethyl cellulose (CMC), polyvinyl alcohol (PVA) and dextrin-based hydrogel membrane were prepared and employed for the sustained release of third-generation oral antibiotic (cefixime). Different proportions of CMC, PVA and dextrin were blended and hydrogel membranes were fabricated via solvent casting method. The prepared membrane was characterized by FTIR, SEM, UV-visible, TGA and swelling analysis. Cefixime drug was incorporated in the CMC/PVA/dextrin matrix and drug release was investigated. The sustained release of the tested drug (cefixime) was investigated and the drug was released in 120 min in the phosphate-buffered saline (PBS) solution. The antibacterial activity of the prepared membrane was promising against Proteus vulgaris, salmonella typhi, Escherichia coli and Bacillus subtilis strains. The swelling capabilities, thermal stability and non-toxic nature of the prepared CMC/PVA/dextrin membrane could have potential applications for cefixime drug in delivery in a controlled way for the treatment of infectious diseases.

A two-step activation mechanism enables mast cells to differentiate their response between extracellular and invasive enterobacterial infection

Mast cells localize to mucosal tissues and contribute to innate immune defense against infection. How mast cells sense, differentiate between, and respond to bacterial pathogens remains a topic of ongoing debate. Using the prototype enteropathogen Salmonella Typhimurium (S.Tm) and other related enterobacteria, here we show that mast cells can regulate their cytokine secretion response to distinguish between extracellular and invasive bacterial infection. Tissue-invasive S.Tm and mast cells colocalize in the mouse gut during acute Salmonella infection. Toll-like Receptor 4 (TLR4) sensing of extracellular S.Tm, or pure lipopolysaccharide, causes a modest induction of cytokine transcripts and proteins, including IL-6, IL-13, and TNF. By contrast, type-III-secretion-system-1 (TTSS-1)-dependent S.Tm invasion of both mouse and human mast cells triggers rapid and potent inflammatory gene expression and >100-fold elevated cytokine secretion. The S.Tm TTSS-1 effectors SopB, SopE, and SopE2 here elicit a second activation signal, including Akt phosphorylation downstream of effector translocation, which combines with TLR activation to drive the full-blown mast cell response. Supernatants from S.Tm-infected mast cells boost macrophage survival and maturation from bone-marrow progenitors. Taken together, this study shows that mast cells can differentiate between extracellular and host-cell invasive enterobacteria via a two-step activation mechanism and tune their inflammatory output accordingly.

Transcriptional responses of cytokines, immunoglobulin A, and nitric oxide genes in 1-day-old chicks post Salmonella typhimurium infection: An experimental study.

Salmonella has become one of the hazards prevalent foodborne pathogens causing different diseases in chickens. However, Salmonella typhimurium (ST), a nonhost-specific serovar, is a major avian agent that causes severe disturbance in young chicken wellness. The occurrence of Salmonella in chickens and their antimicrobial resistance were explored in this study. In addition, the immune response of 1-day-old broiler chicks, against multidrug resistant (MDR) ST infection, was also assessed at 4 and 24 hours post infection (pi) in the cecum and spleen, representing their mucosal and systemic immune responses, respectively. A total of 375 samples from 130 diseased and apparently healthy broiler and layer chickens were randomly collected for Salmonella isolation, identification, and resistance profile evaluation, from farms and different clinical laboratories. The immune response of 1-day-old broiler chicks, Ross 308, against in-vivo ST infection was ascertained through the evaluation of heterophile phagocytosis and s expression of cytokines, immunoglobulin A and other immune-regulating genes in the cecum and spleen. Twenty-four, 1-day-old nonvaccinated broiler chicks were used and divided into two groups. The chicks in the infected group were orally inoculated with 0.5 ml of 2 × 108 colony forming units (CFU)/ml of MDR ST suspension, while those in the control group were taken nutrient broth. Seven out of 130 (5.38%) examined chickens were positive for Salmonella. All isolates (100%) were resistant to amoxicillin-clavulanic acid (AMC), cefazolin (CZ), cefoxitin (FOX), ciprofloxacin (CIP), nalidixic acid (NA), tetracycline (TE), fosfomycin (FOS), and colistin (CT) with multiple antimicrobial resistances (MARs) index range of 0.72-0.83, where none of them was resistant to meropenem (MEM). The results of immune response revealed that chicks infected with ST showed significantly different phagocytosis percentages and index values compared to controls. According to the real-time quantitative polymerase chain reaction (RT-qPCR) results, the transcription of IL-8, iNOS, IL-18, IgA, and IFN-γ for chicks infected by ST showed a significantly increased trend (p < 0.01) with increasing chicken age and was higher in the cecum than spleen compared to controls (p < 0.05) during 24 hours after infection. The findings indicated a strong mucosal immune response in the chicks after the ST challenge, which reflects humoral and cellular responses. Our insight recommended the occurrence of a natural immune response stimulator at 1 day age to face the infection, and this can prevent the resistance transfer, with efficient control measures.

Inactivation of Salmonella Enteritidis in liquid egg yolk and egg white using bacteriophage cocktails

Salmonella Enteritidis (SE) is a significant global cause of foodborne illness, often linked to egg contamination. This study evaluated the inhibitory effects of eight bacteriophages (phages) against three SE strains isolated from poultry environments. The most effective phages were selected to formulate different phage cocktails, to enhance the efficacy and prolong inhibition. Four phage cocktails were tested at a multiplicity of infection (MOI) of 100 in tryptic soy broth (TSB), and at MOIs of 100 and 1000 in liquid egg white (EW) and egg yolk (EY) with storage at 8 °C for up to 30 days (d). The effectiveness of the phage cocktails varied significantly among bacterial strains, yet all demonstrated significant reductions compared to the positive control in liquid culture (P < 0.05). Similarly, the tested SE strains in both EW and EY showed significant reductions with phage treatments (P < 0.005), although the effectiveness was influenced by the MOI and medium composition. Treating EY proved to be more challenging, with lower magnitudes of reduction and longer treatment durations required, compared to EW. Reductions ranged from 1 to greater than 4 log CFU/mL in EW and EY after 30 d, with consistently higher reductions achieved at MOI 1000. Phage titers decreased initially, but remained stable following SE inoculation in broth and liquid eggs at 8 °C, indicating that lysis from without mechanisms may have contributed to the inhibitory effect. Notably, phages exhibited stronger attachment to SE in EW, which can be attributed to be less viscous nature of EW compared to EY. This study demonstrated that phage applications in both EW and EY effectively reduced SE counts at 8 °C, with no regrowth during long-term storage. These findings contribute to the development of biocontrol methods that enhance food safety and reduce foodborne outbreaks associated with contaminated egg products.