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Salmonella translocate to the gut epithelium via microfold cells lining the follicle-associated epithelium (FAE). How Salmonella localize to the FAE is not well characterized. Here we use live imaging and competitive assays between wild-type and chemotaxis-deficient mutants to show that Salmonella enterica serotype Typhimurium (S. Typhimurium) localize to the FAE independently of chemotaxis in an ex vivo mouse caecum infection model. Electrical recordings revealed polarized FAE with sustained outward current and small transepithelial potential, while the surrounding villus is depolarized with inward current and large transepithelial potential. The distinct electrical potentials attracted S. Typhimurium to the FAE while Escherichia coli (E. coli) localized to the villi, through a process called galvanotaxis. Chloride flux involving the cystic fibrosis transmembrane conductance regulator (CFTR) generated the ionic currents around the FAE. Pharmacological inhibition of CFTR decreased S. Typhimurium FAE localization but increased E. coli recruitment. Altogether, our findings demonstrate that bioelectric cues contribute to S. Typhimurium targeting of specific gut epithelial locations, with potential implications for other enteric bacterial infections.

Enteric pathogens such as Salmonella enterica serovar Typhimurium experience spatial and temporal changes to the metabolic landscape throughout infection. Host reactive oxygen and nitrogen species non-enzymatically convert monosaccharides to alpha hydroxy acids, including L-tartrate. Salmonella utilizes L-tartrate early during infection to support fumarate respiration, while L-tartrate utilization ceases at later time points due to the increased availability of exogenous electron acceptors such as tetrathionate, nitrate, and oxygen. It remains unknown how Salmonella regulates its gene expression to metabolically adapt to changing nutritional environments. Here, we investigated how the transcriptional regulation for L-tartrate metabolism in Salmonella is influenced by infection-relevant cues. L-tartrate induces the transcription of ttdBAU, genes involved in L-tartrate utilization. L-tartrate metabolism is negatively regulated by two previously uncharacterized transcriptional regulators TtdV (STM3357) and TtdW (STM3358), and both TtdV and TtdW are required for the sensing of L-tartrate. The electron acceptors nitrate, tetrathionate, and oxygen repress ttdBAU transcription via the two-component system ArcAB. Furthermore, the regulation of L-tartrate metabolism is required for optimal fitness in a mouse model of Salmonella-induced colitis. TtdV, TtdW, and ArcAB allow for the integration of two cues, i.e., substrate availability and availability of exogenous electron acceptors, to control L-tartrate metabolism. Our findings provide novel insights into how Salmonella prioritizes the utilization of different electron acceptors for respiration as it experiences transitional nutrient availability throughout infection.ImportanceBacterial pathogens must adapt their gene expression profiles to cope with diverse environments encountered during infection. This coordinated process is carried out by the integration of cues that the pathogen senses to fine-tune gene expression in a spatiotemporal manner. Many studies have elucidated the regulatory mechanisms of how Salmonella sense metabolites in the gut to activate or repress its virulence program; however, our understanding of how Salmonella coordinates its gene expression to maximize the utilization of carbon and energy sources found in transitional nutrient niches is not well understood. In this study, we discovered how Salmonella integrates two infection-relevant cues, substrate availability and exogenous electron acceptors, to control L-tartrate metabolism. From our experiments, we propose a model for how L-tartrate metabolism is regulated in response to different metabolic cues in addition to characterizing two previously unknown transcriptional regulators. This study expands our understanding of how microbes combine metabolic cues to enhance fitness during infection.

Acute gastrointestinal infection with intracellular pathogens like Salmonella Typhimurium triggers the release of the proinflammatory cytokine interleukin 1β (IL-1β). However, the role of IL-1β in intestinal defense against Salmonella remains unclear. Here, we show that IL-1β production is detrimental during Salmonella infection. Mice lacking IL-1β (IL-1β -/-) failed to recruit neutrophils to the gut during infection, which reduced tissue damage and prevented depletion of short-chain fatty acid (SCFA)-producing commensals. Changes in epithelial cell metabolism that typically support pathogen expansion, such as switching energy production from fatty acid oxidation to fermentation, were absent in infected IL-1β -/- mice which inhibited Salmonella expansion. Additionally, we found that IL-1β induces expression of complement anaphylatoxins and suppresses the complement-inactivator carboxypeptidase N (CPN1). Disrupting this process via IL-1β loss prevented mortality in Salmonella-infected IL-1β -/- mice. Finally, we found that IL-1β expression correlates with expression of the complement receptor in patients suffering from sepsis, but not uninfected patients and healthy individuals. Thus, Salmonella exploits IL-1β signaling to outcompete commensal microbes and establish gut colonization. Moreover, our findings identify the intersection of IL-1β signaling and the complement system as key host factors involved in controlling mortality during invasive Salmonellosis.

Genetic screening of pools of mutants can reveal genetic determinants involved in complex biological interactions, processes, and systems. We previously constructed two single-gene deletion resources for Salmonella enterica serovar Typhimurium 14028s in which kanamycin (KanR) and chloramphenicol (CamR) cassettes were used to replace non-essential genes. We have now used lambda-red recombination to convert the antibiotic cassettes in these resources into a tetracycline-resistant (TetR) version where each mutant contains a different 21-base barcode flanked by Illumina Read1 and Read2 primer sequences. A motility assay of a pool of the entire library, followed by a single-tube processing of the bacterial pellet, PCR, and sequencing, was used to verify the performance of the barcoded TetR collection. The new resource is useful for experiments with defined subsets of barcoded mutant strains where biological bottlenecks preclude high numbers of founder bacteria, such as in animal infections. The TetR version of the library will also facilitate the construction of triple mutants by transduction. The resource of 6197 mutants covering 3490 genes is deposited at Biological and Emerging Infections Resources (beiresources.org).

Background: Salmonella, a major foodborne pathogen, significantly contributes to global diarrheal diseases. Following infection, the primary manifestation often involves the development of acute gastroenteritis, with only a small subset of serotypes capable of precipitating severe systemic illness, marked by fever, sepsis, or miscarriage. Detailed case reports focusing specifically on urinary tract infections caused by Salmonella Typhimurium have not been documented. Case presentation: A 49-year-old Chinese female presented with significant upper ureteral stricture, with a visible narrow segment of about 3 cm in length on retrograde pyelography. The decision was made to proceed with robotic ureteroplasty with a lingual mucosa graft. Eleven days postoperatively, the patient developed recurrent fever. Bacterial culture of mid-stream urine detected Salmonella typhi twice and Escherichia coli once.The patient was treated with cefoperazone/sulbactam and meropenem successively. Fifty days after surgery, double-J stent removal was performed, and the patient remained afebrile thereafter. Conclusions: This case suggests that urinary tract infection may not be the most crucial factor influencing the survival of autologous tissue grafts. With aggressive antimicrobial therapy according to sensitivity, the survival of autologous tissue grafts remains unaffected. Our case may offer valuable insights on urinary tract infections in the context of autologous tissue transplantation for ureteral augmentation. [ABSTRACT FROM AUTHOR]

Background: Salmonella Typhimurium (S. Typhimurium) is a common pathogenic microorganism and poses a threat to the efficiency of poultry farms. As signaling molecules regulating the interaction between the host and gut microbiota, bile acids (BAs) play a protective role in maintaining gut homeostasis. However, the antibacterial effect of BAs on Salmonella infection in broilers has remained unexplored. Therefore, the aim of this study was to investigate the potential role of feeding BAs in protecting against S. Typhimurium infection in broilers. Methods: A total of 144 1-day-old Arbor Acres male broilers were randomly assigned to 4 groups, including non-challenged birds fed a basal diet (CON), S. Typhimurium-challenged birds (ST), S. Typhimurium-challenged birds treated with 0.15 g/kg antibiotic after infection (ST-ANT), and S. Typhimurium-challenged birds fed a basal diet supplemented with 350 mg/kg of BAs (ST-BA). Results: BAs supplementation ameliorated weight loss induced by S. Typhimurium infection and reduced the colonization of Salmonella in the liver and small intestine in broilers (P < 0.05). Compared to the ST group, broilers in ST-BA group had a higher ileal mucosal thickness and villus height, and BAs also ameliorated the increase of diamine oxidase (DAO) level in serum (P < 0.05). It was observed that the mucus layer thickness and the number of villous and cryptic goblet cells (GCs) were increased in the ST-BA group, consistent with the upregulation of MUC2 gene expression in the ileal mucosa (P < 0.05). Moreover, the mRNA expressions of Toll-like receptor 5 (TLR5), Toll-like receptor 4 (TLR4), and interleukin 1 beta (IL1b) were downregulated in the ileum by BAs treatment (P < 0.05). 16S rDNA sequencing analysis revealed that, compared to ST group, BAs ameliorated the decreases in Bacteroidota, Bacteroidaceae and Bacteroides abundances, which were negatively correlated with serum DAO activity, and the increases in Campylobacterota, Campylobacteraceae and Campylobacter abundances, which were negatively correlated with body weight but positively correlated with serum D-lactic acid (D-LA) levels (P < 0.05). Conclusions: Dietary BAs supplementation strengthens the intestinal mucosal barrier and reverses dysbiosis of gut microbiota, which eventually relieves the damage to the intestinal barrier and weight loss induced by S. Typhimurium infection in broilers. [ABSTRACT FROM AUTHOR]

ABSTRACT Adding plant extracts to sausage and other meat products is very important to improve their quality, safety, and durability. The aim of this study was to evaluate the microbiological properties of beef sausage enriched with roselle (Hibiscus sabdariffa L.) sepal extract. The total content of phenolic and flavonoid compounds in hydroalcoholic extract of Roselle sepals was measured by Folin‐Ciocalteu test and aluminum chloride colorimetric method, and antioxidant activity was measured by DPPH method. Pectin was used to encapsulate the extract. The characteristics of capsules, including particle size, zeta potential, and capsule efficiency were measured. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of extracts were performed by tube dilution method. Microbial tests were performed on sausage treatment samples. Analysis of the obtained data was done using SPSS and Excel software. The study revealed that the total phenolic and flavonoid compounds in the extract were 174.6 mg of gallic acid per gram and 16.8368 mg of quercetin, respectively. The extract displayed the highest antioxidant activity at elevated concentrations. Particle size ranged from 16.833 to 640.534 μm. The zeta potential and capsule efficiency were found to be satisfactory. Compared to the encapsulated extract, the free roselle extract better inhibited Staphylococcus aureus (s. aureus), Salmonella typhimurium (S. typhimurium), and Escherichia coli (E.coli). During storage until day 20, these bacteria did not grow in the free or encapsulated extract groups. However, S. aureus appeared in the free extract group from day 25 and the encapsulated extract group from day 20. Roselle extract has antimicrobial properties that can improve the quality and safety of beef sausage during storage. [ABSTRACT FROM AUTHOR]

ABSTRACT Molecular characteristics of emulsifiers such as their molecular weight (MW) and surface charge, not only affect the stability of the emulsion but also can have an impact on its capacity to either inhibit or promote microbial proliferation. These characteristics can affect the behavior of pathogens such as Salmonella Typhimurium in emulsion systems. The growth and thermal resistance of S. Typhimurium were monitored at different oil content levels (20%, 40%, and 60%) in emulsions stabilized by three whey protein‐based emulsifiers: whey protein isolate (WPI), whey protein hydrolysate (WPH), and a modified WPI with an alteration of charge (WPI+). Our study revealed that emulsifier itself with different MW and surface charge had no effect on bacterial growth and inactivation without oil inclusion (p > 0.05). However, it was found that higher bacterial growth rate at 60% oil content emulsion stabilized with WPI+ (0.65 ± 0.03 log CFU/h) than WPI (0.19 ± 0.04 log CFU/h), which showed the charge of emulsifiers has different effects on microbial dynamics in oil‐in‐water emulsion. Interestingly, WPI+ in emulsions also seemed to convey protection against thermal inactivation of bacteria. These data describe a complex interrelationship between the physicochemical characteristics of the emulsifier and its interacting nature with bacterial cells. They throw even more light on the insight about the importance of a strategic approach toward emulsifier selection in food formulations. This is crucial for the food safety and stability of products. [ABSTRACT FROM AUTHOR]

An ethanol extract of Piper auritum leaves (PAEE) inhibits the mutagenic effect of three food-borne aromatic amines (2-amino-1-methyl-6-phenylimidazo[4,5- b ]pyridine (PhIP); 2-amino-3,8-dimethylimidazo[4,5- f ]quinoxaline (MeIQx); 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (4,8-DiMeIQx)) in the TA98 Salmonella typhimurium strain. Preincubation with MeIQx demonstrated in mutagenesis experiments that inhibition of Cytochrome P450 (CYP), as well as direct interaction between component(s) of the plant extract with mutagens, might account for the antimutagenic observed effect. Gas chromatography/mass spectrometry analysis revealed that safrole (50.7%), α-copaene (7.7%), caryophyllene (7.2%), β-pinene (4.2%), γ-terpinene (4.1%), and pentadecane (4.1%) as the main components (PAEE). Piper extract and safrole were able to inhibit the rat liver microsomal CYP1A1 activity that participates in the amines metabolism, leading to the formation of the ultimate mutagenic/ molecules. According to this, safrole and PAEE-inhibited MeIQx mutagenicity but not that of the direct mutagen 2-nitrofluorene. No mutagenicity of plant extract or safrole was detected. This study shows that PAEE and its main component safrole are associated with the inhibition of heterocyclic amines activation due in part to the inhibition of CYP1A subfamily activity. [ABSTRACT FROM AUTHOR]

The effects of both live and heat‐killed states of Lactiplantibacillus plantarum MWFLp‐182 (L. plantarum MWFLp‐182) isolated from the faeces of long‐lived humans on preventing and relieving Salmonella infection were studied. Evaluation methods included RT‐qPCR measurement of inflammatory factors, histopathological analysis and non‐targeted metabolomics. The results showed that L. plantarum MWFLp‐182 in both states improved body weight loss, organ swelling and ileocolon pathological damage. They also protected against Salmonella infection by regulating key KEGG enrichment pathways and the expression levels of TNF‐a and IL‐6. It is worth noting that the function of live bacteria was better than that of heat‐killed bacteria. [ABSTRACT FROM AUTHOR]

Ensuring food safety measures are essential to minimize the risk of foodborne diseases linked to raw food products. Here, we investigated the efficacy of an innovative approach for the control of Salmonella Typhimurium found in fresh produces. Plasma activated water (PAW) and bacteriophages are emerging effective and valuable alternative methods for microbiological decontamination. The efficacy of PAW and a lytic bacteriophage (109 PFU/mL), both separately and sequentially, against S. Typhimurium in fresh produce was investigated. S. Typhimurium (105–107 CFU/g) were inoculated on lettuce leaves and treated with PAW, S. Typhimurium phage SK-T2 or their combination. PAW or bacteriophage inactivated S. Typhimurium, on lettuce leaves at different initial populations, by 2.90–3.46 or 1.45–3.25 log CFU/g, respectively. After sequential treatments of PAW and bacteriophage, S. Typhimurium populations, initially applied at ~ 105 CFU/g reduced by 4.47 log CFU/g, but when the order of application was changed (i.e., bacteriophage followed by PAW), the combination synergistically decreased the Salmonella numbers below the detection limit of the method used for the enumeration (i.e., < 101 CFU/g). At the high-level inoculum (~ 7 log CFU/g), consecutive treatments of PAW and phage decreased the S. Typhimurium population by 3.28 log CFU/g, and a reduction of 6.20 log CFU/g was achieved after reversing the order of treatment. Regardless of the bacterial inoculum level, sequential applications of bacteriophage and PAW resulted in a higher level of inactivation. This study proved that the bacteriophage–PAW combination constitutes a promising alternative approach to the conventional washing process in fresh produce wash waters in the food industry. [ABSTRACT FROM AUTHOR]

Summary: Pork cutlets are susceptible to cross‐contamination with foodborne pathogens during processing. Food irradiation effectively improves food hygiene by eliminating microbes. We investigated the application of X‐ray irradiation (0.5–10 kGy) on pathogen inactivation (D10 value), quality changes [total aerobic bacteria (TAB), yeast and mould (Y&M), total coliform (TC), colour, thiobarbituric acid reactive substance (TBARS), and pH], and the hydrocarbon profile of pork cutlets. D10 values of Escherichia coli O157:H7, Salmonella typhimurium, Listeria monocytogenes, and Staphylococcus aureus were 0.37, 0.62, 0.52, and 0.36 kGy, respectively. At least 3.10 kGy (5‐D10 values) is required to ensure the microbiological safety of pork cutlets. X‐ray irradiation dose‐dependently reduced all microbial counts, and up to 7 kGy had no adverse effects on colour, TBARS, and pH. Furthermore, 1,7‐hexadecadiene and 8‐heptadecene were potential markers for irradiated pork cutlets. X‐ray irradiation of up to 7 kGy is effective in improving hygiene levels while maintaining pork cutlet quality. [ABSTRACT FROM AUTHOR]

Background: Salmonella, is one of the most important pathogens causing foodborne illnesses worldwide. The early detection of Salmonella is essential to minimize the risk. Herein, the nanoprobe (CS-AuNPs@phage-T156) for Salmonella was obtained by functionalizing cysteamine (CS) stabilized gold nanoparticles (CS-AuNPs) with phage T156. Results: In the presence of target Salmonella, the CS-AuNPs@phage-T156 specifically recognized and adsorbed on the surface of Salmonella, then induced the aggregation of gold nanoparticles and the color changes from red to purple, which enabled visual detection. The concentration of Salmonella was quantified by the absorption ratio (A680/A526), with a wide linear range from 1.2 × 101 CFU/mL to 1.2 × 106 CFU/mL. Under the optimal experimental conditions, this assay possessed a potent selectivity characteristic quickly toward Salmonella, with a detection limit of 12 CFU/mL. Conclusion: This simple and rapid detection method is expected to provides a novel identification element for the detection of Salmonella by intuitive visualization judgment and quantitative analysis. [ABSTRACT FROM AUTHOR]

As cytoplasmic protein complexes that are pivotal for innate immunity, inflammasomes act primarily through the detection of pathogen‐ or danger‐associated molecular patterns. Nucleotide oligomerisation domain‐like receptor family and caspase activation recruitment domain‐containing protein 4 (NLRC4) inflammasomes identify and eliminate intracellular pathogens, a process contingent on the ligand‐recognition capabilities of neuronal apoptosis inhibitory proteins (NAIPs). Upon detection of specific molecules indicative of intracellular infection, NAIPs discern distinct pathogenic components and subsequently transmit signals to NLRC4, thus initiating their activation and triggering an inflammatory response. However, the mechanisms underlying NLRC4 inflammasome remain unclear. In this study, we elucidated the critical role of ATG16L2 in activating the NLRC4 inflammasome. ATG16L2‐deficient macrophages exhibited reduced NLRC4 inflammasome activation, characterised by decreased oligomerisation of apoptosis‐associated speck‐like protein containing a CARD and attenuated cleavage of Pro‐caspase‐1, Pro‐IL‐1β and gasdermin D. Co‐immunoprecipitation assays revealed an interaction between ATG16L2 and NAIPs. Furthermore, ATG16L2 enhanced the association between NAIPs and NLRC4 by binding to NAIPs. For ATG16L2‐knockout mice infected with Salmonella typhimurium, pathogen clearance and survival rates markedly decreased. Collectively, our findings suggest that ATG16L2 is a significant modulator of the innate immune system, influencing the activity of the NLRC4 inflammasome and the host's defensive response to intracellular pathogens. [ABSTRACT FROM AUTHOR]

Cryptophytes are a promising source of bioactive compounds that have not been fully explored. This research investigated the antimicrobial activity of total phenolic compounds (TPC) and exopolysaccharides (EPS) extracted from several cryptophytes against a range of harmful foodborne bacteria and fungi. To measure the minimum inhibitory concentration (MIC) value, the broth microdilution method was used. In the antibacterial evaluation of TPC, the MIC ranged between 31.25 and 500 µg/mL, while for the antifungal activity test, it varied from 31.25 to 125 µg/mL. In the antibacterial activity test of EPS, the MIC values ranged from 125 to 1,000 µg/mL, whereas in the antifungal susceptibility test, it ranged between 62.5 and 1,000 µg/mL. The most resistant pathogen against TPC was Escherichia coli, while Campylobacter jejuni was the most susceptible. In the case of EPS, the most resistant pathogen was Salmonella Typhimurium, while Aspergillus versicolor exhibited the highest susceptibility. Overall, in terms of antimicrobial activity, TPC was more effective than EPS. Finally, the tolerance level (TL) for TPC and EPS was =4 in all tested samples, indicating their bactericidal/fungicidal mechanism of action. In conclusion, TPC and EPS isolated from cryptophytes demonstrated remarkable antimicrobial properties and ability to fully eradicate pathogens, and could be considered as natural preservatives in the food industry. [ABSTRACT FROM AUTHOR]

Reuterin, a mixture of different forms of 3-hydroxypropanal (3-HPA), including HPA hydrate and HPA dimer, is an antimicrobial compound converted from glycerol by Lactobacillus reuteri and other strains. Although its antimicrobial function may be related to its interaction with thiol groups, its temperature stability and effect on the gut environment remain unclear. The present study evaluated the antimicrobial effects and activity of reuterin against Escherichia coli and Salmonella typhimurium. Utilization of a reliable in vitro gut microbiome fermentation system revealed that reuterin has a modulatory effect on the gut microbial community. Reuterin treatment completely inhibited H2 and NH3 production in the gut and significantly enhanced the synthesis of branched short-chain fatty acids. 16s rRNA sequencing indicated that reuterin promoted the growth of Proteobacteria and Bacteroidetes in the in vitro system and significantly modulated gut microbiota composition. [ABSTRACT FROM AUTHOR]

The main objective of this study was to evaluate the biological activities of Anabasis setifera extract, including its antimicrobial, anticancer, and antioxidant properties. In the current study, Anabasis setifera leaves extract was evaluated for antimicrobial, anticancer, antioxidant activities and phytochemical analyses. Ethyl acetate extract of Anabasis setifera (EA-AS) exhibited promising antimicrobial activity toward Escherichia coli, Staphylococcus aureus, Salmonella typhimurium, Bacillus subtilis, Candida albicans, Aspergillus brasiliensis, Aspergillus fumigatus with MICs 62.5, 125, 62.5, 31.25, 62.5, 125 and 125 μg/mL respectively. Moreover, EA-AS showed anticancer activity at safe concentrations, where IC50 were 36.4 and 44 μg/mL toward Hep-G2 and MCF-7 cancerous cell lines. EA-AS was found to contain 55 significant compounds identified through gas chromatography mass spectrophotometry (GCMS). The most abundant compounds were 1,4-dimethoxy-6,7,8,9-tetrahydro-5-benzocycloheptenone (26.04%), hexa-2,4-diyn-1-ylbenzene (8.40%), dihydrobenzo[b]fluoranthene (6.10%), ethanone, 1-[2,3-dihydro-2-(1-methylethenyl)-5-benzofuranyl (6.10%), and valerenol (4.08%). GC mass analysis confirmed the antioxidant properties of AS by detecting several compounds with antioxidant activity, including hexa-2,4-diyn-1-ylbenzene, nerolidol, spathulenol, -naphthalenem ethanol, decahydro-4-trimethyl-8-methylene, hexadecenoic acid, tremetone, desmethoxyencecalin, heptadecyn-1-ol, thunbergol, hexadecanol, dotriacontane, taylorione, ligulatin, retinoic acid, and falcarinol. The analysis of EA-AS reveals that it is a rich source of valuable phytochemicals: total Phenolic Content: a promising 4,264 μg/mL /, suggesting substantial biological and pharmacological potential. Total tannin content: 391.17 μg/mL, indicating potential applications in industries like nutraceuticals, pharmaceuticals, and cosmetics. Total flavonoid content exceptionally high at 5,163 μg/mL, while the total alkaloid content measured 1,036.26 μg/mL. Additionally, EA-AS demonstrated antioxidant activity with an EC50 of 30.6 μg/mL. In conclusion, the comprehensive analysis of the EA-AS reveals its immense potential as a rich source of valuable phytochemicals with diverse bioactivities, warranting further in-depth studies to unlock its full pharmaceutical and commercial prospects. Our results suggest substantial biological and pharmacological prospects for EA-AS as a promising antimicrobial, anticancer, and potent antioxidant. [ABSTRACT FROM AUTHOR]

The eukaryotic Glucose Induced Degradation/C-Terminal to LisH (GID/CTLH) complex is a highly conserved E3 ubiquitin ligase involved in a broad range of biological processes. However, a role of this complex in host anti-microbial defenses has not been described. We exploited Mycobacterium tuberculosis (Mtb) induced cytotoxicity in macrophages in a FACS based CRISPR genetic screen to identify host determinants of intracellular Mtb growth restriction. Our screen identified 5 (GID8, YPEL5, WDR26, UBE2H, MAEA) of the 12 predicted members of the GID/CTLH complex as determinants of intracellular growth of both Mtb and Salmonella serovar Typhimurium. We show that the anti-microbial properties of the GID/CTLH complex knockout macrophages are mediated by enhanced GABAergic signaling, activated AMPK, increased autophagic flux and resistance to Mtb induced necrotic cell death. Meanwhile, Mtb isolated from GID/CTLH knockout macrophages are nutritionally starved and oxidatively stressed. Our study identifies the GID/CTLH complex activity as broadly suppressive of host anti-microbial responses against intracellular bacterial infections. Here, Simwela et al. perform a whole genome CRISPR/Cas9 screen on Mycobacterium tuberculosis infected primary macrophages and identify the GID/CTLH complex as a modulator of host cell physiology capable of promoting bacterial survival and growth. [ABSTRACT FROM AUTHOR]

Serotonergic neurons in the central nervous system control behavior and mood, but knowledge of the roles of serotonergic circuits in the regulation of immune homeostasis is limited. Here, we employ mouse genetics to investigate the functions of enteric serotonergic neurons in the control of immune responses and find that these circuits regulate IgA induction and boost host defense against oral, but not systemic Salmonella Typhimurium infection. Enteric serotonergic neurons promote gut-homing, retention and activation of intestinal plasmacytoid dendritic cells (pDC). Mechanistically, this neuro-immune crosstalk is achieved through a serotonin-5-HT receptor 7 (HTR7) signaling axis that ultimately facilitates the pDC-mediated differentiation of IgA+ B cells from IgD+ precursors in the gut. Single-cell RNA-seq data further reveal novel patterns of bidirectional communication between specific subsets of enteric neurons and lamina propria DC. Our findings thus reveal a close interplay between enteric serotonergic neurons and gut immune homeostasis that enhances mucosal defense. Roles of serotonergic neurons in immune homeostasis and host defense are largely unexplored. Here the authors show that in mice enteric serotonergic neurons regulate immune homeostasis and host defense against Salmonella infection by orchestrating gut homing and activation of plasmacytoid dendritic cell, ultimately promoting the differentiation of protective IgA+ B cells. [ABSTRACT FROM AUTHOR]

Campylobacter and invasive non-typhoidal Salmonella (iNTS) are among the most common causative agents of gastroenteritis worldwide. As of now, no single combination licensed vaccine is available for public health use against both iNTS and Campylobacter species. Outer-membrane vesicles (OMVs) are nanoscale proteoliposomes released from the surface of gram-negative bacteria during log phase and harbor a variety of immunogenic proteins. Based on epidemiology of infections, we formulated a novel trivalent outer membrane vesicles (TOMVs)-based vaccine candidate against Campylobacter jejuni (CJ), Salmonella Typhimurium (ST) and Salmonella Enteritidis (SE). Isolated OMVs from CJ, ST and SE were combined in equal ratios for formulation of TOMVs and 5 µg of the developed vaccine candidate was used for intraperitoneal immunization of adult BALB/c mice. Immunization with TOMVs significantly activated both the humoral and cellular arm of adaptive immune response. Robust bactericidal effect was elicited by TOMVs immunized adult mice sera. TOMVs immunization induced long-term protective efficacy against CJ, ST and SE infections in mice. The study illustrates the ability of TOMVs-based combination immunogen in eliciting broad-spectrum protective immunity against prevalent Campylobacter and iNTS pathogens. According to the findings, TOMVs can work as a potent combination-based acellular vaccine candidate for amelioration of Campylobacter and iNTS-mediated gastroenteritis. [ABSTRACT FROM AUTHOR]

High-pressure processing (HPP) is a promising technology for increasing the shelf life of food, with minimal effects on the nutritional or sensory quality. However, there has been a concern that high-oil-content foods may protect food pathogens in HPP, and that HPP can affect the quality of lipids. We inoculated Listeria monocytogenes and Salmonella Typhimurium into 34% and 54% oil-content pesto sauce, processed them either with HPP (600 MPa, 4 min) or thermal processing (82 °C, 5 min), and analyzed bacteria counts, pH, GC-MS (Terpene compounds), the time–kill kinetic study, and lipid oxidation value for 60 days in refrigerating storage (5 ± 2 °C). Our findings show that HPP significantly reduced the number of bacteria (more than 4-log) compared to thermal processing or non-processing. Additionally, we discovered terpene compounds (highest-level terpene: L-linalool, eugenol, and 1,8-cineol) in pesto oil that exhibit antimicrobial activity. Different oil content did not have any significant effect on bacteria levels. Regarding chemical results, all samples were of acceptable quality, and the processes did not show any negative effect on lipid oxidation (Peroxide and P-Anisidine value under 10 meq per kilogram of oil). In conclusion, our study indicates that HPP is a suitable method for high-oil-content pesto sauce. In addition, functional compounds naturally present in pesto may contribute to maintaining its microbial and chemical quality. [ABSTRACT FROM AUTHOR]

The aim of this study was to examine the biological activity and probiotic properties of lactic acid bacteria (LAB) isolated from sweet potato stalk kimchi (SPK). Various LAB and Bacillus spp. are active in the early stages of the fermentation of kimchi made from sweet potato stalk. Four strains of LAB were identified, including SPK2 (Levilactobacillus brevis ATCC 14869), SPK3 (Latilactobacillus sakei NBRC 15893), SPK8 and SPK9 (Leuconostoc mesenteroides subsp. dextranicum NCFB 529). SPK2, SPK3, SPK8, and SPK9 showed 64.64–94.23% bile acid resistance and 78.66–82.61% pH resistance. We identified over 106 CFU/mL after heat treatment at 75 °C. Four strains showed high antimicrobial activity to Escherichia coli and Salmonella Typhimurium with a clear zone of >11 mm. SPK2 had the highest antioxidative potentials, higher than the other three bacteria, with 44.96 μg of gallic acid equivalent/mg and 63.57% DPPH scavenging activity. These results demonstrate that the four strains isolated from sweet potato kimchi stalk show potential as probiotics with excellent antibacterial effects and may be useful in developing health-promoting products. [ABSTRACT FROM AUTHOR]

Among innovative food technologies, ultrasounds have demonstrated physical damages (provided by frequency and intensity factors) on bacterial structures while determining the microbiological stabilization of many foodstuffs. This study tested the efficacy of the thermosonication process on 16 Salmonella typhimurium strains belonging to the academic biobank (isolated from swine slaughterhouses). All strains were exposed to focused ultrasounds, generated by the Waveco® system (Milan, Italy), with the following settings: 40 KHz coupled with 80 W at different 5 min intervals starting from 5 to 15 ones, and focusing on two different temperatures: 40 °C and 50 °C. After each treatment, all strains were directly plated onto count agars immediately (t0) and after 24 h (t24) of storage at refrigerated temperature. The results showed bacterial reductions by prolonging the sonication treatments until 15 min (i.e., 50 °C for 15 min reduced of 2.16 log CFU/gr the initial loads). In the present in vitro study, the most considerable decrease was observed after 24 h. It meant that Salmonella strains were lethally damaged at the wall level, confirming the ultrasound bactericidal effect on loads. The present in vitro scientific investigation demonstrates the practical bactericidal effects of thermosonication, highlighting promising applications at the industry level for food microbial stabilization and shelf-life prolongation. [ABSTRACT FROM AUTHOR]

Three novel 4‐aryl‐2‐(2‐(4‐fluorophenyl)‐2‐oxoethyl)‐6‐phenylpyridazin‐3(2H)‐one derivatives were synthesized, and their chemical structures were established based on 1H‐NMR, 13C‐NMR, ESI‐HRMS, and FT‐IR spectral data. Their in vitro antibacterial activities were evaluated against Staphylococcus aureus [MRSA] (Gram‐positive isolate), Escherichia coli and Salmonella typhimurium (Gram‐negative isolates), and Pseudomonas aeruginosa and Acinetobacter baumannii (Gram‐negative nonfermenter isolates), and the results were compared with the standard drug "Amikacin". The results showed that one of the compounds has good antibacterial activity against P. aeruginosa (MIC = 11.57 µM) and E. coli (MIC = 23.14 µM). In order to gain molecular insights into the compounds, their reactivity properties were studied using density functional theory (DFT), and molecular docking was carried out for the compounds with E. coli MurB protein receptors (2Q85). [ABSTRACT FROM AUTHOR]

Pathogens detection is a crucial measure in the prevention of foodborne diseases. This study developed a novel multicolor colorimetric assay to visually detect Salmonella Typhimurium (S. Typhimurium), by utilizing the etching process of gold nanorods (AuNRs) with TMB2+. The strategy involved the construction of nanozyme by assembling magnetic covalent organic framework (MCOF) with aptamer-conjugated AuNPs (Apt-AuNPs), exhibiting remarkable peroxidase-like activity to catalyze the oxidation of TMB/H2O2 and inducing the etching of AuNRs. The presence of S. Typhimurium could inhibit this process, resulting in the generation of vivid colors. The multicolor colorimetric assay could specifically determine S. Typhimurium from 102 to 108 CFU mL−1 in 60 min with visual detection limit of 102 CFU mL−1, and instrumental detection limit of 2.3 CFU mL−1. Moreover, detecting S. Typhimurium in chicken, milk, pork and lettuce samples has shown promise in practical applications. [ABSTRACT FROM AUTHOR]

Bovine salmonellosis is an important contagious disease that is usually endemic to a particular region. the present study aimed to study the clinical cases, diagnostic tools and the specific gene responsible for the pathogenesis in Basrah, Iraq. The study was conducted on 2- 4 years-old (88) native cow breeds from November 2022 to July 2023. Nearly, 38.63% of cows were positive for Salmonella by clinical signs, culture and confirmed using polymerase chain reaction (PCR). Diseased cows suffered from diarrhea, dehydration and pyrexia. Hematological analysis indicated leukocytosis, neutrophilia and significantly higher erythrocyte sedimentation rate and clotting factor indices. Results also indicated hypoglycemia, decreased total protein, and increased blood urea nitrogen. Fecal samples (1gm) were taken directly from the rectum of all suspected animals and transmitted immediately to the diagnostic lab in a cooling box. The PCR amplification and sequencing results of the 16S rRNA gene from Salmonella isolates revealed that 10 out of 19 isolates exhibited distinct bands of approximately 1500 bp corresponding to the Salmonella enterica serovar typhimurium. The results of cdtb and pltB genes amplification exhibited clear bands of approximately 819 bp and 518 bp, respectively. According to the phylogenetic analysis, our local isolates represented a close relationship to the other published strains in the NCBI database. In conclusion, bovine salmonellosis is considered a serious disease due to its negative effects on the infected animals of all ages. Therefore, early elimination and control of this disease could be the right way to protect farm animals. [ABSTRACT FROM AUTHOR]

Fenugreek, black cumin, and garlic are herbs that are rich sources of antimicrobial and nutritional bioactive compounds. This study investigated the influence of herbs on the rheological characteristics of dough and the quality parameters of bread. The antimicrobial activity of the herbs was determined against the pathogens at 50 and 100 mg/mL concentrations. Maximum zone inhibition was measured for fenugreek against Salmonella typhimurium (16.5 mm) at 100 mg/mL concentration. Dough development time and departure time increased with the increase in concentration of the herbs. Farinograph quality number (FQN), mixing tolerance index, and stability varied significantly with the treatments such as physio-chemical, structural, textural, and sensory parameters at herbs concentration 0.25, 0.5, and 0.75%, respectively. Ash, fiber, fat, protein content, firmness, and total phenolic content were increased with treatments. However, a decrease in moisture content and total plate count was observed due to the addition of herbs. The microstructure revealed that the addition of fenugreek significantly improves the bread structure. The sensory profile of bread was not affected with the addition of herbs at a concentration of 0.50%. [ABSTRACT FROM AUTHOR]

The impact of pulsed light treatment (PLT) on natural microbiota and inoculated microbes such as Salmonella Typhimurium, Bacillus cereus, and Aspergillus flavus on red chilies was investigated. Sequential drying did not completely inactivate the aerobic mesophiles and yeast and mold count. Hence, PLT (0.53–2.59 J cm−2) was employed as a decontamination technology on red chilies. PLT resulted in 8 log reduction of inoculated microorganisms on chilies at 2.59 J cm−2. The microbial inactivation kinetics followed Weibull distribution (R2 > 0.97) with β value of 1.1, 1.2, and 1.5 for S. Typhimurium, B. cereus, and A. flavus, respectively. Changes in structure and composition of cell components were identified by SEM and FTIR analysis. After PLT, phenolics, antioxidants, flavonoids, and capsaicinoids were better retained but a significant change in ascorbic acid and carotenoid's content was observed. Hence, PL can be a potential technology for decontamination of fresh and dried chilies along with maximum retention of bioactives. [ABSTRACT FROM AUTHOR]

Simple Summary: Subunit vaccines based on peptides as minimal immunogenic epitopes/antigens can induce highly specific immune responses without triggering adverse reactions. Here, Salmonella enterica serovar Typhimurium peptides were identified in the immunopeptidome of porcine macrophages genotyped for different class II molecules that were stimulated with the bacteria. Immunopeptidomics and mass spectrometry approaches were used for peptide identification, along with two predictors for immunoinformatics analysis. Differences between individuals were observed concerning length, elution preferences, and elution predictions. Thirty-one bacterial peptides were identified; outer membrane protein-A and chaperonin GroEL protein had the highest number of peptides. Since proteins having the highest number of peptides have been suggested as candidate antigens for vaccines, all the bacterial peptides identified can be considered promising candidate epitopes upon determining their immunogenicity. Salmonellosis is a zoonotic infection that has a major impact on human health; consuming contaminated pork products is the main source of such infection. Vaccination responses to classic vaccines have been unsatisfactory; that is why peptide subunit-based vaccines represent an excellent alternative. Immunopeptidomics was used in this study as a novel approach for identifying antigens coupled to major histocompatibility complex class II molecules. Three homozygous individuals having three different haplotypes (Lr-0.23, Lr-0.12, and Lr-0.21) were thus selected as donors; peripheral blood macrophages were then obtained and stimulated with Salmonella typhimurium (MOI 1:40). Although similarities were observed regarding peptide length distribution, elution patterns varied between individuals; in total, 1990 unique peptides were identified as follows: 372 for Pig 1 (Lr-0.23), 438 for Pig 2 (Lr.0.12) and 1180 for Pig 3 (Lr.0.21). Thirty-one S. typhimurium unique peptides were identified; most of the identified peptides belonged to outer membrane protein A and chaperonin GroEL. Notably, 87% of the identified bacterial peptides were predicted in silico to be elution ligands. These results encourage further in vivo studies to assess the immunogenicity of the identified peptides, as well as their usefulness as possible protective vaccine candidates. [ABSTRACT FROM AUTHOR]

A highly specific and sensitive rapid two-signal assay was developed for the detection of Salmonella typhimurium in foods of animal origin. The invA gene of Salmonella was used as the biorecognition element and recombinase-assisted amplification (RAA) technology for signal amplification. By utilizing the specific recognition and efficient trans-cleavage activity of CRISPR/Cas12a, point-of-care testing (POCT) for S. typhimurium was achieved via lateral flow strips (LFS) and personal glucometer (PGM) biosensors as dual signal readout systems, with sensitivities of 33 CFU/mL and 20 CFU/mL, respectively. Users can select the appropriate test system on the basis of specific application requirements: LFSs are ideal for rapid onsite screening, whereas glucometer biosensors offer precise quantitative determination. This approach simplifies the use of large instruments and overcomes site constraints, demonstrating good accuracy and applicability in animal-derived samples, with significant potential for the detection of other pathogens and for use in restricted environments. [ABSTRACT FROM AUTHOR]

An innovative method is introduced based on the combination of label-free surface-enhanced Raman scattering with advanced multivariate analysis. This technique allows both quantitative and qualitative assessment of Salmonella typhimurium and Escherichia coli on eggshells. Using silver nanocubes embedded in polydimethylsiloxane, we consistently achieved Raman spectra of bacteria. The stability of the Ag NCs@PDMS substrate is confirmed using rhodamine 6G over 30 days under standard conditions. Principal component analysis (PCA) effectively distinguishes between S. typhimurium and E. coli spectra. Partial least squares regression (PLS) models were developed for quantitative determination of bacteria on egg surfaces, yielding accurate results with minimal error. The S. typhimurium model achieves Rc2 = 0.9563 and RMSEC = 0.601 in calibration, and Rv2 = 0.9113 and RMSEV = 0.907 in validation. Similarly, the E. coli model achieves Rc2 = 0.9877 and RMSEC = 0.322 in calibration, and Rv2 = 0.9606 and RMSEV = 0.579 in validation. Recoveries validate PLS predictions by inoculating egg surfaces with varying bacterial amounts. Our study demonstrates the feasibility of SERS-PLS for quantitative determination of S. typhimurium and E. coli on eggshells, promising enhanced food safety protocols. [ABSTRACT FROM AUTHOR]

Four synthetic Schiff bases (PSB1 [(E)-2-(((4-aminopyridin-3-yl)imino)methyl)-4,6-dibromophenol], PSB2 [(E)-2-(((4-aminopyridin-3-yl)imino)methyl)-4,6-diiodophenol], PSB3 [(E)-2-(((4-aminopyridin-3-yl)imino)methyl)-4-iodophenol], and PSB4 [(E)-2-(((4-aminopyridin-3-yl)imino)methyl)-4-chloro-6-iodophenol]) were fully characterized. These compounds exhibit an intramolecular hydrogen bond between the hydroxyl group of the phenolic ring and the nitrogen of the azomethine group, contributing to their stability. Their antimicrobial activity was evaluated against various Gram-negative and Gram-positive bacteria, and it was found that the synthetic pyridine Schiff bases, as well as their precursors, showed no discernible antimicrobial effect on Gram-negative bacteria, including Salmonella Typhi (and mutant derivatives), Salmonella Typhimurium, Escherichia coli, and Morganella morganii. In contrast, a more pronounced biocidal effect against Gram-positive bacteria was found, including Bacillus subtilis, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus haemolyticus. Among the tested compounds, PSB1 and PSB2 were identified as the most effective against Gram-positive bacteria, with PSB2 showing the most potent biocidal effects. Although the presence of reactive oxygen species (ROS) was noted after treatment with PSB2, the primary mode of action for PSB2 does not appear to involve ROS generation. This conclusion is supported by the observation that antioxidant treatment with vitamin C only partially mitigated bacterial inhibition, indicating an alternative biocidal mechanism. [ABSTRACT FROM AUTHOR]

Organic compounds with antibacterial and antiparasitic properties are gaining significance for biomedical applications. This study focuses on the solvent-free synthesis (green synthesis) of 1,4-naphthoquinone or 2,3-dichloro-1,4-naphthoquinone with different phenylamines using silica gel as an acid solid support. The study also includes in silico PASS predictions and the discovery of antibacterial and antiparasitic properties of phenylaminonaphthoquinone derivatives 1–12, which can be further applied in drug discovery and development. These activities were discussed in terms of molecular descriptors such as hydrophobicity, molar refractivity, and half-wave potentials. The in vitro antimicrobial potential of the synthesized compounds 1–12 was evaluated against a panel of six bacterial strains (three Gram-positive: Staphylococcus aureus, Proteus mirabilis, and Enterococcus faecalis; and three Gram-negative bacteria: Escherichia coli, Salmonella typhimurium, and Klebsiella pneumoniae). Six compounds (1, 3, 5, 7, 10, and 11) showed better activity toward S. aureus with MIC values between 3.2 and 5.7 μg/mL compared to cefazolin (MIC = 4.2 μg/mL) and cefotaxime (MIC = 8.9 μg/mL), two cephalosporin antibiotics. Regarding in vitro antiplasmodial activity, compounds 1 and 3 were the most active against the Plasmodium falciparum strain 3D7 (chloroquine-sensitive), displaying IC50 values of 0.16 and 0.0049 μg/mL, respectively, compared to chloroquine (0.33 μg/mL). In strain FCR-3 (chloroquine-resistant), most of the compounds showed good activity, with compounds 3 (0.12 μg/mL) and 11 (0.55 μg/mL) being particularly noteworthy. Additionally, docking studies were used to better rationalize the action and prediction of the binding modes of these compounds. Finally, absorption, distribution, metabolism, excretion, and toxicity (ADMET) predictions were performed. [ABSTRACT FROM AUTHOR]

Cervical cancer is the fourth most common malignancy in women globally. Chemotherapies, targeted therapies, and immunotherapies in the treatment of cervical cancer are usually accompanied by effective and adverse effects. Therefore, finding other efficient and accurate molecular targets remains essential to improve the treatment benefits of cervical cancer patients. MCPIP1 (monocyte chemoattractant protein-induced protein 1) is a kind of endonuclease with a CCCH zinc finger domain and a PilT-N-terminal (PIN) domain, and its function in cervical cancer is unknown. We found that MCPIP1 inhibits cell proliferation and promotes cell apoptosis of cervical cancer. Additionally, MCPIP1 suppresses mRNA and protein expression of the apoptotic inhibitor XIAP by decreasing its mRNA stability. Mechanically, MCPIP1 binds to the XIAP mRNA via its CCCH zinc finger domain and degrades the XIAP mRNA via the endonuclease activity coming from its PIN domain. Our study clarifies that MCPIP1 promotes cervical cancer cell apoptosis by suppressing the expression of XIAP, thereby impeding cervical cancer progression. Moreover, targeted delivery of MCPIP1 with engineered Salmonella typhimurium leads to tumor growth retardation in the HeLa xenograft tumor model in mice. Therefore, our study may provide a theoretical basis for formulating clinical treatment strategies for cervical cancer. [ABSTRACT FROM AUTHOR]

Inertial microfluidics is essential for separating particles and cells, enabling numerous biomedical applications. Despite the simplicity of spiral microchannels, the lack of predictive models hampers real‐world applications, highlighting the need for cost‐effective computational tools. In this study, four novel data fitting models are developed using linear and power regression analyses to investigate how flow conditions influence particle behaviors within spiral microchannels. These models are rigorously tested under two different flow rates, focusing on a smaller particle representing Salmonella Typhimurium and a larger particle representing bacterial aggregates, aiming for effective separation and detection. A critical parameter, the sheath‐to‐sample flow rate ratio, is either interpolated or extrapolated using the microchannel's aspect ratios to predict particle separation. The models show strong agreement with experimental data, underscoring their predictability and efficiency. These insights suggest that further refinement of these models can significantly reduce research and development costs for advanced inertial microfluidic devices in biomedical applications. This work represents a crucial step towards establishing a robust computational framework, advancing inertial microfluidics towards practical biomedical applications. [ABSTRACT FROM AUTHOR]

It is thought to be risk-free, environmentally benign, and safe for biological processes to produce zinc oxide nanoparticles from renewable resources. This study examined Cassia javanica's ability to create ZnONPs. The generated ZnONPs were analyzed using a variety of techniques, such as TEM, FTIR spectroscopy, UV-Vis spectroscopy, and XRD analysis. The antibacterial potential of ZnONPs has been investigated using both Agar well diffusion and microtitreplate (MTP) methods. One method used to evaluate ZnONPs' capacity to scavenge free radicals at different concentrations was the DPPH method. The permanent zinc oxide (ZnO) shape and the naturally occurring crystal structure of ZnONPs were validated by the XRD data. ZnONPs showed antibacterial activity with MICs of 31.7 μg/mL toward Bacillus subtilis, 62.5 μg/mL for Salmonella typhimurium, Escherichia coli while Clostridium sporogenes and Bacillus pumilus was 125μg/mL. Furthermore, ZnONPs demonstrated a range of antibiofilm activities toward Staphylococcus aureus (MRSA). ZnONPs showed an intriguing antioxidant capacity, achieving IC50 of 109.3 μg/ml μg/mL. Additionally, ZnONPs demonstrated low toxic effect on Vero cell with IC50 154.01 μg/mL as well as possible anticancer action when applied to the carcinoma cell lines HepG2 with IC50 of 47.48 μg/mL. Furthermore, ZnONPs at 62.5 μg/mL had a promising antiviral impact against HSV1 and COX B4, with antiviral activities of 75.4% and 65.8%, respectively. [ABSTRACT FROM AUTHOR]

With advancements in DNA amplification research, isothermal amplification technology has emerged as an attractive method for detecting target DNA. Here, we describe primer generation-rolling circle amplification (PG-RCA) as an isothermal amplification method for detecting Escherichia coli O157:H7, Salmonella Typhimurium, Bacillus cereus, and Listeria monocytogenes. To improve PG-RCA sensitivity, the concentrations of the reaction components, dNTPs, phi29 DNA polymerase, and circular probes were optimized; the optimized conditions were applied to detect each target bacterium. A pair of forward and reverse circular probes that hybridized to the sense and anti-sense target genes was used in PG-RCA, exhibiting target selectivity. PG-RCA, which generated additional primers simultaneously with linear RCA and comprised multiple reaction cycles, resulted in higher accumulation of amplified DNA products than did linear RCA within the same reaction period. The threshold time (Tt) for each target gene concentration was determined based on the threshold value set in the amplification plot for PG-RCA, and a linear correlation between the Tt value and genomic DNA concentration was proven for each of the four bacteria. The PG-RCA-based assay could be applied to gene-based detection of various microorganisms and may be a useful isothermal amplification method for replacing traditional PCR methods. [ABSTRACT FROM AUTHOR]

Salmonellosis represents a significant economic and public health concern for the poultry industry in Africa, leading to substantial economic losses due to mortality, reduced productivity, and food safety problems. However, comprehensive information on the burden of poultry salmonellosis at the continental level is scarce. To address this gap, a systematic review and meta-analysis were conducted to consolidate information on the prevalence and circulating serotypes of poultry salmonellosis in African countries. This involved the selection and review of 130 articles published between 1984 and 2021. A detailed systematic review protocol was structured according to Cochrane STROBE and PRISMA statement guideline. From the 130 selected articles from 23 different African countries, the overall pooled prevalence estimate (PPE) of poultry salmonellosis in Africa was found to be 14.4% (95% CI = 0.145–0.151). Cameroon reported the highest PPE at 71.9%. The PPE was notably high in meat and meat products at 23%. The number of research papers reporting poultry salmonellosis in Africa has shown a threefold increase from 1984 to 2021. Salmonella Enteritidis and Typhimurium were the two most prevalent serotypes reported in 18 African countries. Besides, Salmonella Kentucky, Virchow, Gallinarum, and Pullorum were also widely reported. Western Africa had the highest diversity of reported Salmonella serotypes (141), in contrast to southern Africa, which reported only 27 different serotypes. In conclusion, poultry salmonellosis is highly prevalent across Africa, with a variety of known serotypes circulating throughout the continent. Consequently, it is crucial to implement strategic plans for the prevention and control of Salmonella in Africa. RESEARCH HIGHLIGHTS The pooled sample prevalence of poultry salmonellosis in Africa is high (14.4%). The highest PPE was recorded in meat and meat products. Salmonella serotypes of zoonotic importance were found in all sample types. Salmonella Enteritidis and Typhimurium are common serotypes spreading in Africa. [ABSTRACT FROM AUTHOR]

Salmonella Typhimurium is an invasive gastrointestinal pathogen for both humans and animals. To investigate the genetic framework and diversity of S. Typhimurium, a total of 194 S. Typhimurium isolates were collected from patients in a tertiary hospital between 2020 and 2021. Antimicrobial susceptibility testing was used to confirm the resistance phenotype. Whole-genome sequencing and bioinformatics analysis were performed to determine the sequence type, phylogenetic relationships, resistance gene profiles, Salmonella pathogenicity island (SPI) and the diversity of the core and pan genome. The result showed that 57.22% of S. Typhimurium isolates were multidrug resistant and resistance of total isolates to the first-line drug ciprofloxacin was identified in 60.82%. The population structure of S. Typhimurium was categorized into three lineages: ST19 (20.10%, 39/194), ST34-1 (47.42%, 92/194) and ST34-2 (40.65%, 63/194), with the population size exhibiting increasing trends. All lineages harbored variety of fimbrial operons, prophages, SPIs and effectors that contributed to the virulence and long-term infections of S. Typhimurium. Importantly, ST34-1 lineage might potentially be more invasive due to the possession of SPI1-effector gene sopE which was essential for the proliferation, internalization and intracellular presence of S. Typhimurium in hosts. Multiple antimicrobial resistance genes were characteristically distributed across three lineages, especially carbapenem genes only detected in ST34-1&2 lineages. The distinct functional categories of pan genome among three lineages were observed in metabolism, signaling and gene information processing. This study provides a theoretical foundation for the evolved adaptation and genetic diversity of S. Typhimurium ST19 and ST34, among which ST34 lineages with multidrug resistance and potential hypervirulence need to pay more attention to epidemiological surveillance. [ABSTRACT FROM AUTHOR]

The present study focused on the quantitative determination of active compounds, antioxidant and antibacterial activities of leaf extracts of different Sonneratia species. The tested species including Sonneratia alba (SA), Sonneratia caseolaris (SC), and Sonneratia ovata Backer (SO) were collected from Can Gio Mangrove Forest, Vietnam. The methanol extract of SA leaves was found to contain the highest content of total flavonoids, total phenolics and gallic acid (4344.16 ± 51.78 μg/g dry mass). The highest luteolin‐7‐O‐glucoside and luteolin amounts (5963.28 ± 59.77 and 234.33 ± 10.41 μg/g dry mass, respectively) were found in the methanol extract of SC leaves according to HPLC‐DAD analysis. The methanol extracts of all three species exhibited strong DPPH radical scavenging activity while the ethanol extracts provided strong antibacterial activity against Staphylococcus aureus, Salmonella typhimurium, and Escherichia coli. By fractionating the extracts over a SiO2 column using CHCl3: EA: MeOH gradients, the fractions containing high purity luteolin and luteolin‐7‐glucoside from SC and high purity luteolin from SO were collected. Moreover, all fractions demonstrated significant DPPH free radical scavenging ability but the fraction enriched in gallic acid showed the highest activity. These findings show that SA is a promising source of gallic acid while SC can be potentially used for the extraction and isolation of luteolin‐7‐glucoside. [ABSTRACT FROM AUTHOR]

The current study aimed to evaluate the antibacterial efficacy of black seed oil (BSO) and its nanoemulsion (BSO-NE) against two foodborne pathogens, Listeria monocytogenes and Salmonella Typhimurium, during the processing and storage of yoghurt. Additionally, the sensory properties of the resulting yoghurt were assessed. Firstly, BSO-NE was prepared and characterized using zeta-sizer and Transmission electron microscopy (TEM). Then, the antibacterial activity and minimum inhibitory concentration (MIC) were determined via a resazurin-based microtiter dilution assay. Fresh buffalo's milk was inoculated with L. monocytogenes and Salmonella Typhimurium (1×105 CFU/ml) with the addition of either BSO or BSO-NE. The inoculated milk was then used for yoghurt making. The organoleptic properties and bacterial load of the obtained yoghurt were evaluated during storage in comparison to a control group. This study elucidated that BSO can be successfully produced in a nanoemulsion form, exhibiting acceptable particle size, stability, and morphology. The results demonstrated a strong antibacterial activity of BSO-NE (3.125 mg/mL) against both L. monocytogenes and Salmonella Typhimurium compared to the control and pure oil groups. Notably, complete eradication of L. monocytogenes and Salmonella Typhimurium was observed in yoghurt formulated with BSO-NE on the 8th day of storage. Interestingly, the antibacterial effect of BSO and BSO-NE on Salmonella Typhimurium was approximately similar throughout the experiment. Moreover, the results indicated that the addition of BSO-NE to yoghurt enhanced its organoleptic properties. Overall, these findings suggest that yoghurt can be fortified with BSO-NE to prevent bacterial contamination by L. monocytogenes and Salmonella Typhimurium in the dairy industry. [ABSTRACT FROM AUTHOR]

Antimicrobial drug resistance caused by Salmonella Typhimurium (S. Typhimurium) is a noteworthy issue causing morbidity and mortality linked to potential outbreaks throughout the world. The current study aimed to synthesis chitosan, iron oxide nanoparticles (Ch-Fe3O4 NPs), tannic acid-Ch-Fe3O4 (T-Ch-Fe3O4) NPs for the loading of florfenicol, compare the antibacterial efficiency of the free florfenicol and the loaded one against florfenicol-resistant strains of S. Typhimurium. Also, the in-vitro bio-toxicity and viability effect of those materials were assessed. It was shown that florfenicol was successfully loaded onto the surface of tannic acid-Ch-Fe3O4 NPs. This was determined through measurement of hydrodynamic sizes and zeta potential (+0.120±0.02, -0.372±0.072 and +0.07±0.03 mV for Ch-Fe3O4 NPs, T-Ch-Fe3O4 NPs, and Florfenicol NPs respectively) and confirmed by FT-IR analysis. Concerning the antibacterial effect, loading florfenicol with T-Ch-Fe3O4 NPs resulted in superior antibacterial efficacy against the resistant S. Typhimurium strain. This was evident by a decrease in the minimum inhibitory concentration (MIC) from 31.25 µg/ml for florfenicol to 15.62 µg/ml for the loaded florfenicol NPs. This finding suggests that florfenicol NPs possess enhanced antibacterial efficacy compared to the free drug, highlighting the potential of nanotechnology in addressing the challenge of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Foodborne pathogens, such as Salmonella, are major factors that pose significant threats to global food safety and public health. Salmonella typhimurium is a prominent serotype contributing to non-typhoidal salmonellosis, which is a prevalent foodborne illness affecting humans and animals. Bacteriophages are considered one of the most environmentally friendly biocontrol agents, particularly in the food industry, owing to their high specificity and high safety. However, the emergency of phage-resistant mutants limits the biocontrol effect of phage treatment, leading to the requirement for a high diversity of lytic phages. Therefore, the study isolated and characterized two novel lytic Salmonella bacteriophages (SPYS_1 and SPYS_2) targeting S. typhimurium ATCC14028 and evaluated their effectiveness in reducing the contamination rates for milk and chicken tenders. Morphological and genomic analyses indicated that Salmonella phages SPYS_1 and SPYS_2 are novel species classified under the genus Skatevirus and the genus Berlinvirus, respectively. Both phages exhibited high stability across a broad range of thermal and pH conditions. The one-step growth curve result suggested that both phages had a short adsorption time and a large burst size in a single lytic cycle. The phage SPYS_1 demonstrated a noteworthy inhibition effect on the growth of S. typhimurium ATCC14028 in milk, resulting in a ~2-log reduction within the 2 to 4 h range. Overall, both phages have shown significant potential for application in food safety in the future. [ABSTRACT FROM AUTHOR]

Salmonella typhimurium and Listeria monocytogenes are relevant foodborne bacterial pathogens which may cause serious intoxications and infectious diseases in humans. In this study, a sensitive immunochromatographic analysis (ICA) for the simultaneous detection of these two pathogens was developed. For this, test strips containing two test zones with specific monoclonal antibodies (MAb) against lipopolysaccharides of S. typhimurium and L. monocytogenes and one control zone with secondary antibodies were designed, and the double-assay conditions were optimized to ensure high analytical parameters. Prussian blue nanoparticles (PBNPs) were used as nanozyme labels and were conjugated with specific MAbs to perform a sandwich format of the ICA. Peroxidase-mimic properties of PBNPs allowed for the catalytic amplification of the colorimetric signal on test strips, enhancing the assay sensitivity. The limits of detection (LODs) of Salmonella and Listeria cells were 2 × 102 and 7 × 103 cells/mL, respectively. LODs were 100-fold less than those achieved due to the ICA based on the traditional gold label. The developed double ICA was approbated for the detection of bacteria in cow milk samples, which were processed by simple dilution by buffer before the assay. For S. typhimurium and L. monocytogenes, the recoveries from milk were 86.3 ± 9.8 and 118.2 ± 10.5% and correlated well with those estimated by the enzyme-linked immunosorbent assay as a reference method. The proposed approach was characterized by high specificity: no cross-reactivity with other bacteria strains was observed. The assay satisfies the requirements for rapid tests: a full cycle from sample acquisition to result assessment in less than half an hour. The developed ICA has a high application potential for the multiplex detection of other foodborne pathogens. [ABSTRACT FROM AUTHOR]

There is an increasing need to develop alternative antimicrobials to replace currently used antibiotics. Phytochemicals, such as essential oils, have garnered significant attention in recent years as potential antimicrobials. However, the mechanisms underlying their bactericidal activities are not yet fully understood. In this study, we investigated the bactericidal activity of eugenol oil against Salmonella enterica serovar Typhimurium (S. Typhimurium) to elucidate its mechanism of action. We hypothesized that eugenol exerts its bactericidal effects through the production of reactive oxygen species (ROS), which ultimately leads to cell death. The result of this study demonstrated that the bactericidal activity of eugenol against S. Typhimurium was significantly (p < 0.05) mitigated by thiourea (ROS scavenger) or iron chelator 2,2′-dipyridyl, supporting the hypothesis. This finding contributes to a better understanding of the killing mechanism by eugenol oil. [ABSTRACT FROM AUTHOR]

The cuticle covering the outer surface of an eggshell functions as both a physical and chemical barrier against invading microorganisms. Contamination of eggs by microbial pathogens progresses in four stages: bacterial attachment to the egg surface, penetration through the cuticle and eggshell, multiplication within the underlying membranes, and the final stage of contaminating the egg contents. Therefore, it is important to study bacterial count at the first point of contact, i.e., on the surface of the eggs. In this study, we have evaluated the impact of differences in cuticle quality (due to egg washing and hen age) on bacterial load. We compared bacterial adherence on the eggshell surface of white eggs which were either washed (graded) or unwashed (ungraded), collected from Lohmann laying hens of different ages: early (24–28 weeks), mid-lay (44–48 weeks), and late (66–70 weeks). We aimed to determine the impact of hen age and egg washing on differences in cuticle quality and bacterial adherence. Our results indicate that hen age (up to 70 weeks) and commercial egg washing do not significantly impact bacterial adherence on eggshell surfaces. We have developed a novel method using green fluorescent protein (GFP)-expressing Salmonella typhimurium to estimate adherence of bacteria to the eggshell surface, with independent measurement of autofluorescence to quantitate cuticle deposition. S. typhimurium were localized, adhering to cracks visible on the outer cuticle in ungraded eggs, indicating that egg-associated pathogens usually enter the egg interior either through respiratory pores in eggshells or through shell micro-cracks. The results of this study can be utilized to optimize innovative methods for predictive microbiology in order to achieve egg safety. [ABSTRACT FROM AUTHOR]

The rapid and sensitive detection of Salmonella typhimurium in food matrices is crucial for ensuring food safety. This study presents the development of an ultrasensitive electrochemical biosensor using surface-modified bacterial cellulose (BC) integrated with polypyrrole (Ppy) and reduced graphene oxide (RGO), further functionalized with immobilized S. typhimurium-specific phage particles. The BC substrate, with its ultra-fibrous and porous structure, was modified through in situ oxidative polymerization of Ppy and RGO, resulting in a highly conductive and flexible biointerface. The immobilization of phages onto this composite was facilitated by electrostatic interactions between the polycationic Ppy and the negatively charged phage capsid heads, optimizing phage orientation and enhancing bacterial capture efficiency. Morphological and chemical characterization confirmed the successful fabrication and phage immobilization. The biosensor demonstrated a detection limit of 1 CFU/mL for S. typhimurium in phosphate-buffered saline (PBS), with a linear detection range spanning 100 to 107 CFU/mL. In real samples, the sensor achieved detection limits of 5 CFU/mL in milk and 3 CFU/mL in chicken, with a linear detection range spanning 100 to 106 CFU/mL, maintaining high accuracy and reproducibility. The biosensor also effectively discriminated between live and dead bacterial cells, demonstrating its potential in real-world food safety applications. The biosensor performed excellently over a wide pH range (4–10) and remained stable for up to six weeks. Overall, the developed BC/Ppy/RGO–phage biosensor offers a promising tool for the rapid, sensitive, and selective detection of S. typhimurium, with robust performance across different food matrices. [ABSTRACT FROM AUTHOR]

Salmonella is the main bacterial pathogen that causes foodborne disease, particularly in developing countries. Nontyphoidal Salmonella (NTS) include Enteritidis and Typhimurium as the most prevalent strains which are one of the significant causes of acute gastroenteritis in children. Therefore, identifying the most predominant serovars, types of common contaminated food, and paying attention to their antibiotic resistance are the main factors in the prevention and control strategy of salmonellosis. This study was undertaken to evaluate the prevalence rate of serovars, the biofilm formation, antimicrobial resistance (AMR) status, and phenotypic virulence factors of Salmonella strains isolated from diarrhea samples in some cities of Iran. A total of 40 (10.41%) Salmonella isolates were recovered from 384 diarrhea samples processed and the most common serovar was Salmonella serovar Typhimurium (82.5). Also, all isolates belonging to serovar Typhimurium showed more virulence factors compared to other serovars. The isolates showed a high resistance rate to ampicillin (95%) and nalidixic acid (87.5%), while a low resistance rate was found for chloramphenicol (2.5%). Moreover, significant variances in the capacity of biofilm formation were found between different Salmonella serotypes. The resistance of NTS to extant choice drugs is a potential public health problem. Constant monitoring of AMR pattern and virulence profile of NTS serovars is suggested for the prevention of salmonellosis in humans. [ABSTRACT FROM AUTHOR]

In this investigation, the bio‐activity and biosafe potential of thiosemicarbazonates of nickel(II) as well as that of ligands are described. Reactions of nickel(II) acetate with a series of thiosemicarbazones, {R1(py)C2=N3‐N2(H)‐C1(=S)‐NHR; R1=py, Me, H and Ph; R=H, Me, Et, Ph}, led to the loss of acidic hydrazinic N2(H) protons, and resulted in the formation of six‐coordinated complexes of stoichiometry, [Ni(N,N,S−L)2], where anionic N,N,S−L=py2tsc‐NHR, 1–4; apytsc‐NHR, 5–8; pytsc‐NHR, 9–12 and bpytsc‐NHR, 13–16. All these complexes have been characterized using analytical data, IR and UV‐visible spectroscopy and ESI‐mass spectrometry. The single crystal X‐ray crystal structure of several complexes {2(Me), 7(Et), 11(Et), 12(Ph), 13(H),14(Me) and 15(Et)} were also determined. Thio‐ligands coordinate to nickel(II) through pyridyl nitrogen‐N4, azomethine nitrogen‐N3 and thiolato sulfur atoms, resulting in the formation of homoleptic complexes with octahedral structures. Complexes 1–16 and uncoordinated thiosemicarbzones, were evaluated for their antimicrobial activity, in terms of ZOI, MIC, time kill assay and in vitro percent cell viability MTT assay, against clinical isolate methicillin‐resistant Staphylococcus aureus (MRSA), Staphylococcus aureus (MTCC740), Klebsiella pneumonia (MTCC109), Salmonella typhimurium (MTCC1251), and Candida albicans (MTCC227), a yeast. Both the ligands and their metal complexes exhibited moderate to high bioactivity, and were found to be biosafe with high cell viability, 88–92 %, in several cases. [ABSTRACT FROM AUTHOR]

Objective: Persister cells are a specific subset of bacteria capable of surviving exposure to lethal doses of antibiotics, leading to antibiotic therapy failures and infection relapses. This research explores the utilization of drug repositioning to target the Lon protease in Salmonella Typhimurium. Method: In this study, FDA-approved drugs sourced from the Drug Bank database were screened to identify existing pharmaceuticals with the potential to combat the Lon protease. The formation of persister cells in the presence of antibiotics, as well as the combination of antibiotics with potential Lon protease inhibitors, was examined. Furthermore, the expression of type II toxin-antitoxin system genes was analyzed to enhance our comprehension of the inhibitors' effects. Result: Molecular docking analysis revealed that Diosmin and Nafcillin exhibited strong binding affinity to the Lon protease. Molecular dynamics simulation trajectories analysis demonstrated that the interaction of these ligands with the enzyme did not induce instability; rather, the enzyme's structure remained stable. Combinations of ceftazidime and ciprofloxacin with either Nafcillin or Diosmin led to significant reductions in bacterial cell counts. Furthermore, the effectiveness of these combinations, when compared to antibiotics alone, highlighted the substantial impact of Nafcillin and Diosmin in reducing type II TA system gene expression. Conclusion: These findings suggest promising prospects for developing novel therapeutic approaches targeting persister cells to mitigate treatment failures in Salmonella infections. [ABSTRACT FROM AUTHOR]