Caffeine holds promise for applications in food safety due to its antioxidant and antibacterial properties. Given rising antimicrobial resistance, its natural antimicrobial potential is valuable for controlling foodborne pathogens and reducing reliance on synthetic preservatives. This study aimed to explore caffeine as an alternative to control Salmonella biofilms in fruit juice substrates. S. Enteritidis and S. Typhimurium biofilm were developed in Brain Heart Infusion (BHI) broth (control), grape and apple juice, before and after caffeine application. Biofilm inhibition was quantified by crystal violet staining, exopolysaccharide (EPS) production, and visualization through Confocal and Scanning Electron Microscopy. Swimming motility assays assessed caffeine's impact on bacterial motility. Both strains formed biofilms in the tested juices. The Minimum Inhibitory Concentration (MIC) of caffeine was 9.37 mM ml-1 for S. Typhimurium and 18.75 mM ml-1 for S. Enteritidis. Biofilm inhibition was observed for treatments before and after caffeine application, with varying levels depending on the matrix. EPS production and inhibition were higher in biofilms formed in grape and apple juices compared to the control (BHI). Sub-inhibitory concentrations of caffeine reduced motility in both strains. These findings suggest that caffeine may be a promising approach to control Salmonella biofilms in the food industry.

The contamination of fresh produce, including blueberries, with pathogenic microorganisms poses a significant public health risk. This study assessed the efficacy of alkyltrimethylammonium bromide (ATMB), a quaternary ammonium compound, in reducing microbial loads on blueberries. The antimicrobial activity of ATMB was tested against Escherichia coli O157:H7, Salmonella enterica, and Listeria monocytogenes at varying concentrations (750 and 1500 ppm) and exposure times (1, 5, and 10 min). Results showed that reductions in microbial loads were dependent on both the concentration of ATMB and exposure time. The greatest reductions of 6.7 log CFU ml-1 (where CFU stands for colony-forming units) for Escherichia coli O157:H7, 5.1 log CFU ml-1 for S. enterica, and 4.9 log CFU ml-1 for L. monocytogenes were observed at 1500 ppm for 10 min. These findings demonstrate the potential of ATMB as an effective antimicrobial agent for the decontamination of blueberries, thereby enhancing food safety and protecting consumers.

Antimicrobial resistance among Staphylococcus species, including multidrug-resistant and biofilm-forming strains, poses a critical threat to global health, demanding innovative therapeutic solutions. In this context, this study explores the antimicrobial and antibiofilm potential of the aquatic plant Hydrocleys nymphoides as a promising alternative. Extracts from the plant's leaves and roots were obtained using solvents of increasing polarity and tested against five key pathogenic Staphylococcus species: S. aureus, S. epidermidis, S. haemolyticus, S. pseudintermedius, and S. coagulans. The hexane extract of H. nymphoides leaves showed the most notable activity, with inhibition zones of 9-17 mm and minimum inhibitory concentrations (MICs) as low as 0.8 mg/ml for certain strains. Subinhibitory concentrations of the extract significantly reduced biofilm formation in most strains, with reductions up to 46.9%. Gas chromatography-mass spectrometry revealed bioactive compounds such as linoleic acid, n-hexadecanoic acid, 9-octadecenal, eicosane, and tetratriacontane, known for their antimicrobial and antibiofilm properties. Although cytotoxicity was observed at concentrations near the MIC, lower concentrations were non-toxic, indicating potential for controlled therapeutic applications. These findings underscore the biotechnological value of H. nymphoides as a sustainable source of antimicrobial agents against multidrug-resistant Staphylococcus. This work emphasizes the critical role of phytotherapy in combating the escalating antimicrobial resistance crisis.

Goat milk is a superior nutritional source for human beings. It possesses probiotic, prebiotic, immunomodulatory actions and inhibits the adherence of pathogenic bacteria. The microbiota of raw goat milk is a rich source of novel bacteriocin-producing lactic acid bacteria (LAB). LAB were evaluated for their probiotic attributes, i.e. acid, bile tolerance, and safety assessment. The antimicrobial activity of LAB isolated from goat milk was assessed against the 10 most common indigenous bacterial pathogens confirmed through 16S rRNA sequencing. The strains LAB GM8 and GM121 showed high tolerance to acidic pH while GM121 showed tolerance to high bile salt concentrations. LAB GM121 showed susceptibility to eight antibiotics which meets the safety requirements regarding phenotypic resistance evaluation. Out of 10, 4 LAB GM8, GM26, GM121, GM122 displayed antibacterial activity against Pseudomonas aeruginosa and Staphylococcus aureus, demonstrating their ability to produce bioactive compounds. So, based on acid, bile tolerance, and safety requirements, it can be concluded that the GM121 has the potential to be explored as probiotic strains in the dairy industry. Bacterial isolates (n-24) were screened for exopolysaccharide production and amylase activity; Bacillus paralicheniformis GM75 showed good exopolysaccharide production and antimicrobial activity while Bacillus cereus GM56 showed maximum amylase activity.

Breast cancer has emerged as the leading cause of global cancer incidence, surpassing lung cancer. Accumulating evidence suggests that probiotics exhibit inhibitory effect on breast cancer progression, highlighting the need to identify gut flora-derived probiotics with potential anti-breast cancer properties. Here, we investigated the effect of the cell-free supernatant of Clostridium leptum (ClCFS) on breast cancer cells by methyl thiazolyl tetrazolium (MTT) assay. Untargeted metabolomics analysis was employed to characterize metabolite alterations in ClCFS. Furthermore, the core targets were predicted by the protein-protein interaction network and the signaling pathways were enriched by the Kyoto Encyclopedia of Genes and Genomes analysis. Our findings demonstrated that ClCFS inhibited the proliferation of breast cancer cells and that the main metabolite of ClCFS might be acetylcarnitine. Utilizing network pharmacological analysis, we identified apoptosis-related signaling pathways as the principal mechanisms underlying ClCFS activity. Furthermore, five core targets of STAT3, IL-1β, BCL2, CASP3, and ESR1 were identified. This study elucidates the main bioactive constituent and the potential targets of ClCFS against breast cancer. It provides a new understanding of the pharmacological activity of ClCFS in breast cancer treatment.

Newcastle Disease (ND) is a significant viral disease affecting poultry worldwide, with outbreaks persisting despite extensive vaccination efforts. This study characterizes a novel NDV strain, RT40, isolated from poultry farms in northeast Iran for the first time. RT40, classified as genotype VII, subgenotype VII.1.1, has a genome length of 15 192 nucleotides, with six genes, 12 UTRs, and five IGS, consistent with class II NDVs. Pathogenicity tests confirm its virulent velogenic nature, differentiating it from other known strains. Phylogenetic analysis points to a Western origin, suggesting possible cross-border transmission with Iraqi strains, highlighting the strain's relevance for NDV surveillance. Virus isolation, RNA sequencing, and cross-border transmission analysis were performed, revealing that RT40 shares a high nucleotide sequence identity with Iraqi strains, further supporting cross-border transmission concerns. The findings underscore the need for genotype-specific vaccines tailored to circulating strains. The genome, deposited in GenBank (accession ON184061), serves as an essential resource for future research and vaccine development. By emphasizing whole-genome sequencing, this study aims to improve NDV evolution tracking and enhance understanding of NDV diversity. This work calls for targeted vaccine strategies to effectively manage NDV spread, ensuring the sustainability of the poultry industry.

Foodborne pathogens present a significant public health concern where fresh produce is a key agricultural product. Rapid and sensitive detection methods are essential to ensure the safety of such produce. This study aimed to develop and optimize a modified multiplex polymerase chain reaction (mPCR) assay, which incorporates enhancements to conventional PCR, for the simultaneous detection of Escherichia coli, Salmonella, and Listeria monocytogenes in fresh produce. The specificity of each primer pair was validated using 15 strains, confirming 100% accurate detection of pathogenic strains without cross-reactivity. Since no false positives were observed, the assay demonstrated 100% precision, highlighting its reliability in distinguishing target pathogens. The sensitivity of the mPCR assay was demonstrated through serial dilutions, detecting Salmonella down to 10 fg µl-1, L. monocytogenes to 100 fg µl-1, and E. coli to 1 pg µl-1. The mPCR assay was then successfully applied to romaine lettuce and kale, demonstrating its effectiveness in detecting pathogens in mixed samples inoculated at varying concentrations (109-101 CFU ml-1). Kale exhibited greater sensitivity, detecting pathogens at lower levels, while romaine lettuce also provided consistent detection. This study highlights the potential of mPCR for enhancing food safety by providing rapid and sensitive pathogen detection in fresh produce.

Sleep is an essential homeostatic process that undergoes dynamic changes throughout the lifespan, with distinct life stages predisposed to specific sleep pathologies. Similarly, the gut microbiome also varies with age, with different signatures associated with health and disease in the latest decades of life. Emerging research has shown significant cross-talk between the gut microbiota and the brain through several pathways, suggesting the microbiota may influence sleep, though the specific mechanisms remain to be elucidated. Here, we critically examine the existing literature on the potential impacts of the gut microbiome on sleep and how this relationship varies across adulthood. We suggest that age-related shifts in gut microbiome composition and immune function may, in part, drive age-related changes in sleep. We conclude with an outlook on the therapeutic potential of microbiome-targeted interventions aimed at improving sleep across adulthood, particularly for individuals experiencing high stress or with sleep complaints.

Cleaning and sanitizing processing equipment are essential for achieving the hygiene standards required in food processing. The term "sanitizing" refers to the inactivation of microorganisms by disinfectants on previously cleaned surfaces and materials. In the food industry, commercial solutions of peracetic acid (PAA) and hydrogen peroxide (HP) show bactericidal and sporicidal activity at room temperature, and are used as disinfectants for food surfaces, fruit, and vegetables, or to sanitize water. The aim of this study was to quantify the impact of commercial solutions of PAA and HP on Bacillus cereus endospores as a function of concentration and temperature, in order to optimize its use. Inactivation kinetics were carried at 5 PAA/HP solution concentration levels and three temperature levels. Weibull model fits on concave inactivation kinetics quantified B. cereus spore resistances. A Bigelow-type model quantified the effects of concentration and temperature on spore resistance Weibull model parameters. The parameters of the Bigelow-type model enable to optimize disinfection treatments, reducing the concentration of PAA/HP solutions used by increasing the treatment temperature, while ensuring the same level of disinfectant efficacy against bacterial spores.

Antimicrobial bioassay-guided isolation of metabolites from fermented rice cultures of Beltraniella portoricensis strain MRH42 yielded known metabolites hymenopsin A (1) and B (2). Their chemical structures were identified by comparison of their nuclear magnetic resonance and high-resolution mass spectra data reported in the literature. Both metabolites were cytotoxic to human tumour cell lines liver (HepG2), colon (HT29), breast (MCF-7), pancreatic (MiaPaca-2), and human epithelial liver cells transformed (THLE2), with ED50 ranging from 45.72 to 76.19 µM for 1 and 14.83-31.66 µM for 2. None of the compounds was active against drug-resistant microbial bacteria. This is the first report of cytotoxic purified metabolites from B. portoricensis. The hymenopsins biosynthesized by B. portoricensis, contribute to the expansion of our understanding of the chemical diversity of fungi native to the subtropical regions of Mexico.

This study designed an in vitro model to test the efficacies of footbath sanitizers for use in dairy processing plants. Efficacies of selected sanitizers for inactivation of mixed-cocktail, dairy-relevant Gram-negative bacteria in model footbaths, in the presence of milk residues, were measured over 7 days against attached and planktonic populations by plate counting and calculating log reductions achieved. A simplified practical table ranked the most to the least effective sanitizers. These included: biguanide/quaternary ammonium compound (QAC) combination > biguanide = accelerated hydrogen peroxide = ethanol/sodium hydroxide combination > phenolic compound = amine amphoteric compounds > amphoteric surfactant > diclosan = chlorine dioxide > enzyme/surfactants combination.

The ability to precisely manipulate the genome of Clostridioides difficile allows for the generation of mutants that can be characterized to unveil critical aspects relating to pathogenesis and virulence. Such findings can ultimately direct the development of antivirulence agents to combat C. difficile infection, by small molecule inhibition. In this review, the mechanisms underpinning the most widely adopted allelic exchange technologies for generating precise genomic mutations in C. difficile, are discussed. The advantages and disadvantages of each system are considered. In addition, a comprehensive review of the application of these tools for the study of C. difficile toxins and genes residing within the toxin loci, is presented.

The current study aimed to isolate and characterize Vibrio cholerae isolated from the Jukskei River, one of the largest Rivers in Johannesburg, South Africa. Water samples collected from the Jukskei River were subjected to culture-based methods for the detection and isolation of V. cholerae. Twenty-four V. cholerae were isolated, confirmed using real-time PCR, and sequenced using the MInION portable nanopore-sequencing device. Reference-based genome assemblies were constructed from the raw reads using the EPI2ME software followed by bioinformatics analysis using the Centre for Genomic Epidemiology website. All the V. cholerae isolates isolated from the Jukskei River were classified as non-O1/non-O139 and none of the isolates harbored the cholera toxin gene, ctxA. All 24 V. cholerae isolates belonged to sequence type 741, virulent genes including toxR, vspD, als, hlyA, makA, and rtxA as well as the Vibrio pathogenicity island 2 were detected amongst the isolates. Antimicrobial resistance genes (parC, varG, and gyrA) were detected in 83% of isolates. Although V. cholerae non-O1/non-O139 are not associated with epidemic cholera they can still cause mild to life-threatening illnesses. Therefore, increased surveillance should be considered to better understand the public health risks to the local community.

This study aimed to characterize Melissa officinalis essential oil (EOMO) from a region in Northeast Brazil and evaluate its antifungal activity against Candida and Cryptococcus neoformans, by analyzing its action mechanism. EOMO was characterized using gas chromatography-mass spectrometry and the main chemical compounds were Geranial (14.10%) and (Z)-Nerolidol (17.75%). The broth microdilution assay was used to determine the minimum inhibitory concentration (MIC) against strains of Candida albicans, Candida parapsilosis, Candida krusei, Candida auris (MIC raging from 256 to 26.7 µg ml-1) and C. neoformans (MIC ranging from 64 to 32 µg ml-1). Flow cytometry and comet assays were employed to investigate EOMO's mechanism of action, which might be related to an increase in the production of reactive oxygen species and damage to fungal DNA. The chemical composition of EOMO from Northeast Brazil showed a higher content of (Z)-Nerolidol and has significant antifungal potential.

Up to 90% of organic matter (OM) in soils and sediments are stabilized and protected against microbial decomposition through organo-mineral interactions, formation of soil aggregates, pH and oxygen availability. In soils and sediment systems OM is associated with mineral constituents promoting carbon persistence and sequestration of which iron (Fe) and manganese (Mn) are crucial components. Under anoxic condition, microbes couple the decomposition of OM to the oxidative/reductive transformation of Fe/Mn minerals leading to carbon greenhouse gas (C-GHG) emissions (i.e., CH4 and CO2). Although these organo-mineral-microbe interactions have been observed for decades, the bio-geochemical mechanisms governing the switch from OM stability toward OM degradation are not fully understood. Interest in this field have been growing steadily given the interest in global warming caused by OM decomposition leading to C-GHG emissions. This review emphasizes the dual role of Fe/Mn minerals in both OM stability and decomposition. Additionally, we synthesize the conceptual understanding of how Fe/Mn minerals govern OM dynamics and the resultant C-GHG emissions via microbial-mediated carbon transformation. We highlight the need for interdisciplinary research to better understand organo-Fe/Mn mineral-microbial interactions to develop management handles for climate mitigation strategies.

Current therapeutic alternatives for the treatment of cryptococcosis are scarce, highly toxic, expensive and difficult to access. Therefore, the aim of this study was to evaluate the anticryptococcal potential of a collection of 27 drugs in vitro against several strains of Cryptococcus neoformans and Cryptococcus gattii. We investigated several parameters to evaluate the antifungal activity of the drugs: determination of minimum inhibitory concentration (MIC), combinatorial effects with fluconazole, kinetics of growth inhibition, post antifungal effect (PAFE) and morphometric analyses at subinhibitory concentrations. Antiparasitics albendazole, fenbendazole, flubendazole, mebendazole and antidepressants duloxetine and paroxetine showed antifungal activity with a MIC of 100 µmol L-1 or less for most strains tested. The results of the zero-interaction power model indicated additive effects for combination of fluconazole with drugs finasteride, hydroxyzine and paroxetine. The combined treatments significantly improved the ability of fluconazole to kill C. neoformans ATCC H99. Same phenomenon occurred in the in vitro PAFE, as the combinations suppressed fungal growth more effectively than fluconazole alone. A significant reduction in capsule size was observed. Screening of the drug collection showed interesting results, with benzimidazoles antiparasitics and serotonin and norepinephrine reuptake inhibitors in foreground. Finasteride, hydroxyzine and paroxetine significantly improved activity of fluconazole in vitro.

Whey from previous production is often used as a natural starter in the technology of traditional cheeses, including bryndza-cheese in Slovakia. Therefore, studying its bacterial community and isolating new potential natural starters is important for improving the characterisitics of final product. Composition of bacterial consortia of fresh and fermented whey in the production of raw ewes´ milk-based bryndza-cheese from 8 small or medium-sized producers was analysed. Culture-based microbiological analysis and culture-independent analysis based on 16S rRNA gene sequencing by MiSeq and MinION were used. Results showed the dominance of lactococci or streptococci, with 3 - 8 log CFU ml-¹ of Lactobacillus sensu lato in all whey samples. Potential natural starters comprising Lacticaseibacillus paracasei/casei, Lactiplantibacillus plantarum, Lentilactobacillus parabuchneri, Lactobacillus helveticus, Lactobacillus diolivorans, Levilactobacillus brevis, Limosilactobacillus fermentum, Lactobacillus delbrueckii, Lactobacillus gasseri and Lentilactobacillus otakiensis were isolated. Coliforms were also present in all samples, with no consistently lower values in fermented whey samples. Some samples contained pseudomonads and/or acinetobacters. Coagulase positive staphylococci were present at relevant levels in samples from 4 producers. The results revealed that whey is a source of natural starters due to the presence of lactobacilli.

The present study employs a metagenomics approach to evaluate microbial communities' ecological functions and potential within the Rajpardi lignite mine of Gujarat, India. Through whole genome shotgun sequencing on the Illumina Miseq platform, we obtained 10,071,318 sequences, which unveiled a diverse and abundant microbial community primarily composed of Proteobacteria, Acidobacteria, and Nitrospirae. Comprehensive taxonomic profiling and gene prediction was carried out using the SqueezeMeta pipline, which highlighted significant contributions to carbohydrate, amino acid, and energy metabolism. The detection of antimicrobial resistance (AMR) and stress resistance genes, such as blaTEM and merA, suggests that these microbes possess the ability to adapt to harsh environmental conditions. Genome binning revealed species such as Acidiphilum sp. 20-67-58, emphasising the nature of these communities as they adapted to an acidic environment. This finding highlights the crucial role of microbes in biogeochemical cycles, emphasizing their potential in bioremediation, pollutant degradation, and ecosystem restoration.

The ability of Salmonella enterica subsp. enterica to persist and form biofilms on different surfaces can constitute a source of food contamination, being an issue of global concern. The objective of this study was to understand the biofilm formation profile of 14 S. enterica strains among different serovars and sources and to evaluate the ability of essential oil (EO) components (carveol, citronellol, and citronellal) to disinfect the biofilms formed on stainless steel and polypropylene surfaces. All the strains were able to form biofilms with counts between 5.34 to 6.78 log CFU cm-2. Then, the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of EO components were evaluated on two selected strains. All compounds inhibited the growth of Salmonella Typhimurium (strain 1; MIC = 800-1000 µg ml-1) and Salmonella Enteritidis (strain 5; MIC = 400-1000 µg ml-1) and only carveol showed bactericidal activity against strains 1 and 5 (MBC = 1200 µg ml-1). Biofilms were exposed to the EO components at 10 × MIC for 30 min and polypropylene surfaces were more difficult to disinfect showing reductions between 0.9 and <1.2 log CFU cm-2. In general, the S. enterica biofilms demonstrated a significant tolerance to disinfection, demonstrating their high degree of recalcitrance on food processing surfaces.

Biofilm and planktonic forms are different kinds of self-protection mechanisms in microorganisms for resistance to adverse environments. The research explored the physicochemical properties, antimicrobial, and antioxidant activities of Lactobacillus delbrueckii (L. delbrueckii) 5E, focusing on the biofilm's stress tolerance. L. delbrueckii 5E's high lactic acid production (709.1 g·L-1) enhances its antimicrobial activity, which is pH-dependent and decreases when the supernatant is adjusted to pH 6.5. The cell-free fermentation supernatant, bacterial suspension, and cell-free extract of L. delbrueckii 5E showed significant scavenging activity against 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, and superoxide anion radicals. L. delbrueckii 5E formed biofilms with a bacterial count of 9.00 × 106 CFU·mL-1 on polyacrylonitrile electrospun membranes. Stress-tolerance tests indicated that the biofilm form of L. delbrueckii 5E exhibited superior survival under high temperatures, osmotic pressures, bile salts, potassium sorbate, and H2O2 exposure. The biofilm consistently released free bacteria, maintaining a stable total colony count of 106 CFU·mL-1. The remarkable antibacterial and antioxidant properties of L. delbrueckii 5E, along with its resilience to harsh environments, establish its potential for applications in the food industry.

Codonopsis radix, Astragalus membranaceus, Platycodon grandiflorus, and Tiger milk mushroom are promising candidates for functional food development. However, their limited bioavailability impedes a challenge to broader applications. In this study, fermentation of these medicine and food homologous (MFHs) materials using Lactobacillus plantarum was optimized to enhance antioxidant activity. Through single-factor experiments and Pareto analysis, the key factors influencing DPPH radical scavenging activity, including the material-to-water ratio, sucrose concentration, and inoculum size, were identified. Response surface methodology and Artificial neural networks were then employed to optimize fermentation parameters. The optimal conditions resulted in a DPPH radical scavenging rate of 95.8%. Furthermore, the levels of bioactive compounds, including polysaccharides, polyphenols, flavonoids, and saponins, were significantly elevated, which enhanced antioxidant activity and may have improved bioavailability. FTIR analysis confirmed the structural changes in the active compounds after fermentation. This study provides valuable insights into developing MFH-based probiotic fermented products, enhancing their nutritional and biological properties.