Search Results (640)

A new Schiff base, (E)-2-(((1,10-phenanthrolin-5-yl)imino)methyl)-4,6-di-tert-butylphenol (Fen-IHB), was designed to incorporate an intramolecular hydrogen bond (IHB) between the phenolic OH and the azomethine nitrogen with the goal of modulating its physicochemical and biological properties. Fen-IHB was synthesized by condensation of 5-amino-1,10-phenanthroline with 3,5-di-tert-butyl-2-hydroxybenzaldehyde and exhaustively characterized by HR-ESI-MS, FTIR, 1D/2D NMR (¹H, ¹³C, DEPT-45, HH-COSY, CH-COSY, D₂O exchange), and UV–Vis spectroscopy. Cyclic voltammetry in anhydrous CH₃CN revealed a single irreversible cathodic peak at −1.43 V (vs. Ag/Ag⁺), which is consistent with the intramolecular reductive coupling of the azomethine moiety. Density functional theory (DFT) calculations, including MEP mapping, Fukui functions, dual descriptor analysis, and Fukui potentials with dual descriptor potential, identified the exocyclic azomethine carbon as the principal nucleophilic site and the phenolic ring (hydroxyl oxygen and adjacent carbons) as the main electrophilic region. Noncovalent interaction (NCI) analysis further confirmed the strength and geometry of the intramolecular hydrogen bond (IHB). In vitro antimicrobial assays indicated that Fen-IHB was inactive against Gram-negative facultative anaerobes (Salmonella enterica serovar Typhimurium and Typhi, Escherichia coli) and strictly anaerobic Gram-positive species (Clostridioides difficile, Roseburia inulinivorans, Blautia coccoides), as any growth inhibition was indistinguishable from the DMSO control. Conversely, Fen-IHB displayed measurable activity against Gram-positive aerobes and aerotolerant anaerobes, including Bacillus subtilis, Streptococcus pyogenes, Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus haemolyticus. Overall, these comprehensive characterization results confirm the distinctive chemical and electronic properties of Fen-IHB, underlining the crucial role of the intramolecular hydrogen bond and electronic descriptors in defining its reactivity profile and selective biological activity. Full article

Mexican culture offers a great variety of traditional maize-based fermented foods that are beneficial for human health. Atole agrio (sour atole), prepared from blue maize (Zea mays) in the state of Veracruz, has been scarcely studied as a potential functional food. The purpose of this study was to select endogenous potentially probiotic lactic acid bacteria (LAB) from freshly fermented blue maize atole agrio. Samples of spontaneously fermented atole agrio were used for the isolation of LAB on MRS agar. The abilities to tolerate acidic pH, bile salts, and sodium chloride, as well as surface hydrophobicity and aggregation capabilities, were used as criteria for probiotic potential. Selected LAB were identified using MALDI-TOF-MS. Finally, safety-related characterizations, such as hemolytic activity and antibiotic susceptibility, were performed. In the initial stages of fermentation, the presence of fungi, yeasts, coliform organisms, and LAB were detected, and in the final fermentation process, where the blue atole agrio reached a pH of 4, 49 isolates of LAB were obtained. Sixteen isolates showed high tolerance to pH 2, and seven of them showed tolerance to 3% bile salts and 4% sodium chloride. The seven isolates were identified as Pediococcus pentosaceus. Although the seven isolates showed low hydrophobicity to hexadecane and chloroform, they had medium autoaggregation and coaggregation with pathogens. The seven isolates showed notable antibacterial properties against Staphylococcus aureus, Salmonella enterica serovar Typhimurium, Escherichia coli, and Listeria monocytogenes, as well as good amylolytic capacity. All the P. pentosaceus strains were non-hemolytic, sensible to clindamycin and resistant to the other 11 antibiotics tested. Only subtle differences were found among the seven isolates, which can be considered potential candidates for probiotics. The freshly fermented blue maize atole agrio can be considered a functional food containing potentially probiotic LAB and the antioxidant phenolic compounds present in blue maize. Full article

The efficacy of peracetic acid (0.05%, 0.5%, and 1%), sodium hypochlorite (0.2%, 0.6%, and 1%), and benzalkonium chloride (0.3%, 1.15%, and 2%) was evaluated against Staphylococcus aureus (ATCC 6538), Salmonella enterica serovar Typhimurium, (ATCC 14028), Enterococcus hirae (ATCC 8043), Pseudomonas aeruginosa (ATCC 9027), Escherichia coli (ATCC 9027), and Listeria monocytogenes (ATCC 35152) using stainless steel discs, following European Committee for Standardization (CEN) guidelines. According to CEN, a sanitizer must achieve at least a 5 Log₁₀ CFU reduction to be considered effective. Peracetic acid at 1% demonstrated the highest inactivation capacity, reducing all tested strains by more than 7 Log₁₀ CFU/mL. P. aeruginosa (ATCC 9027) showed high tolerance to sodium hypochlorite and benzalkonium chloride, with reductions below 2 Log₁₀ CFU/mL even at maximum concentrations. Both sodium hypochlorite and benzalkonium chloride, at their highest tested concentrations, effectively inactivated S. aureus, S. typhimurium, E. hirae, L. monocytogenes, and E. coli, achieving reductions greater than 7 Log₁₀ CFU/mL. Overall, sanitizers were effective only at intermediate or maximum concentrations recommended by the manufacturers, suggesting that minimum label concentrations should be avoided to ensure microbiological control. Full article

The lack of improved sanitation in rural areas of developing countries, including South Africa, exacerbates open defecation, leading to the significant contamination of water sources by human and animal waste. This study aimed to establish the association of Campylobacter jejuni, Salmonella enterica serovar Typhimurium, Shigella flexneri, and Yersinia enterocolitica in open defecation sites and animal waste with the contamination of water sources in Vhembe District, South Africa. A total of 1032 water samples and 111 faecal samples from the Collins Chabane and Thulamela municipalities were analysed using qPCR. Regression models were used to assess associations, with S. Typhimurium (19–60%) and S. flexneri (11–44%) being the most prevalent bacteria in faecal matter and water, showing detection rates of 4–100% and 5–100%, respectively. Strong associations were found during the wet season between faecal waste and water contamination for S. flexneri (R² = 0.7, p = 0.005) and S. Typhimurium (R² = 0.619, p = 0.091). Urgent measures are needed to address the contamination of rural water sources due to open defecation and livestock waste. Full article

This study aimed to develop innovative functional gummy candies enriched with protein hydrolysates derived from porcine liver, enhancing their antioxidant and antimicrobial properties. First, the overall consumer acceptability (OA) was assessed to determine the most suitable combination of gummy matrix components. Selected combinations were then analyzed for antioxidant activity (ABTS•+, DPPH•), antimicrobial effects, microbiological safety, and physicochemical characteristics. The incorporation of liver hydrolysates significantly increased antioxidant capacity. The highest activity was observed in sample GC5Pa24Ag, hydrolyzed with papain for 24 h and formulated with agar, showing ABTS•+ and DPPH• scavenging activities of (67.6 ± 0.98 µmol/g) and (49.14 ± 1.00%), respectively (p ≤ 0.05). Pepsin hydrolyzed samples (GC2Pe3Gl, GC2Pe6Gl, GC2Pe24Gl) exhibited significantly larger inhibition zones against Listeria monocytogenes ATCC 13932, Escherichia coli ATCC 25922, and Salmonella enterica subsp. enterica serovar Typhimurium ATCC 14028 compared to the control (p < 0.05). Among all, GC5Pa24Ag demonstrated the broadest antimicrobial activity, with a 29.0 ± 0.2 mm inhibition zone against all tested pathogens. These findings suggest that porcine liver hydrolysates can be successfully incorporated into confectionery products to create functional gummies with potential health benefits, offering antioxidant protection and antimicrobial effects in a consumer-friendly form. Full article

The growing demand for natural preservatives has driven interest in essential oils (EOs) from medicinal and aromatic plants. This study examines the potential of EOs from six wild populations of Albanian Lamiaceae, specifically Origanum vulgare subsp. hirtum, Thymbra capitata, and Satureja montana species, to be utilized for food conservation, among other possible uses. The EOs were extracted by hydrodistillation, and their chemical profiles were analyzed through GC-MS. DPPH and ABTS assays were performed to evaluate antioxidant activity. The antimicrobial efficacy of the oils was assessed using the broth microdilution method against six common foodborne pathogens: Salmonella enterica serovar Enteritidis, Escherichia coli, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Micrococcus luteus, and one fungus, Candida albicans. The most potent EOs in terms of yield and biological activity, resulting from O. vulgare subsp. hirtum and T. capitata, were encapsulated in oil-in-water emulsions, which were characterized for particle size and zeta potential. The results show that the populations of O.vulgare subsp. hirtum and T. capitata taken in the study belong to carvacrol chemotypes, and their EOs show strong antioxidant activity and are effective against all tested microorganisms. Nanoemulsions prepared with these EOs showed promising stability, indicating their potential as natural preservatives in food applications. Full article

Background/Objectives: The consumption of liquid egg products is rising. While thermal pasteurization improves safety and shelf life, it can affect product quality. Furthermore, egg products continue to cause many foodborne illnesses, especially those caused by Salmonella enterica subspecies enterica serovar Enteritidis (Salmonella Enteritidis). Bacteriophages (or phages) are an effective alternative to specifically fight foodborne bacteria. This study aimed to evaluate (i) the stability of phage vB_SeEM_UALMA_PCSE1 (PCSE1) under different conditions of temperature and pH; (ii) the effect of multiplicity of infection (MOI) and temperature on phage efficacy; (iii) the bactericidal effect of phage PCSE1 against S. Enteritidis in liquid whole eggs compared to thermal pasteurization; and (iv) the effect of both treatments on the physicochemical and functional properties of liquid whole eggs. Methods: For this, stability tests, bacterial growth inhibition assays in culture media and liquid eggs, and physicochemical and functional analyses were conducted. Results: Phage PCSE1 was (i) stable at pH 7 and 8, and at 4, 25, and 37 °C for 56 days; (ii) effectively prevented S. Enteritidis growth in TSB (reduction of 1.8, 4.5, and 4.5 log colony-forming units (CFU)/mL at 4, 10, and 25 °C, respectively, relative to the bacterial control); (iii) controlled S. Enteritidis in liquid whole eggs at 25 °C (reduction of 5.8 log CFU/mL relative to the bacterial control) comparable to pasteurization (reduction of 5.2 log CFU/mL); and (iv) preserved eggs’ properties, contrarily to pasteurization. Conclusions: These findings suggest PCSE1 is a promising strategy to fight S. Enteritidis in liquid egg products, though further studies on shelf-life are needed. Full article

Salmonella serovar identification typically requires multiple enrichment steps using selective media, consuming considerable time and resources. This study presents a rapid, culture-independent method leveraging artificial intelligence (AI) to classify Salmonella serovars from rich hyperspectral microscopy data. Five serovars (Enteritidis, Infantis, Kentucky, Johannesburg, 4,[5],12:i:-) were analyzed from samples prepared using only sterilized de-ionized water. Hyperspectral data cubes were collected to generate single-cell spectra and RGB composite images representing the full microscopy field. Data analysis involved two parallel branches followed by multimodal fusion. The spectral branch compared manual feature selection with data-driven feature extraction via principal component analysis (PCA), followed by classification using conventional machine learning models (i.e., k-nearest neighbors, support vector machine, random forest, and multilayer perceptron). The image branch employed a convolutional neural network (CNN) to extract spatial features directly from images without predefined morphological descriptors. Using PCA-derived spectral features, the highest performing machine learning model achieved 81.1% accuracy, outperforming manual feature selection. CNN-based classification using image features alone yielded lower accuracy (57.3%) in this serovar-level discrimination. In contrast, a multimodal fusion model combining spectral and image features improved accuracy to 82.4% on the unseen test set while reducing overfitting on the train set. This study demonstrates that AI-enabled hyperspectral microscopy with multimodal fusion can streamline Salmonella serovar identification workflows. Full article

Contaminated poultry is one of the major sources of food-borne non-typhoidal Salmonella (NTS). The aim of this study was to evaluate the presence of Salmonella along the slaughter process in low- and high-throughput poultry abattoirs in South Africa and to determine their characteristics. Samples were collected from 500 chicken carcass rinsates at various processing stages in three abattoirs. Salmonella detection and identification was conducted in accordance with the ISO 6579 methodology. NTS serotyping was performed with serotype-specific PCRs. The Kirby–Bauer disk diffusion method was used to determine antimicrobial resistance in Salmonella. PCR was used to analyze thirteen antimicrobial genes and four virulence genes. Salmonella spp. was detected in 11.8% (59/500; CI: 9.5–15) of the samples tested. The predominant serovars were Salmonella Enteritidis (n = 21/59; 35.59%) and Salmonella Typhimurium (n = 35; 59.32%). Almost all Salmonella isolates were susceptible to all tested antimicrobials except three. Despite the low resistance to tetracyclines at the phenotypic level, approximately half of the strains carried tetA genes, which may be due to “silent” antimicrobial resistance genes. Diverse virulence genes were detected among the confirmed NTS serotypes. We found a predominance of S. Enteritidis and S. Typhimurium from chicken carcasses with diverse virulence and resistance genes. As we detected differences between the slaughterhouses, an in-depth study should be performed on the risk of Salmonella in low- and high-throughput abattoirs. The integrated monitoring and surveillance of NTS in poultry is warranted in South Africa to aid in the design of mitigation strategies. Full article

Background: Salmonella is a bacterium that causes foodborne infections. This study characterized two strains isolated from cheese and beef in Brazil using whole-genome sequencing (WGS). Objectives: We evaluated their antimicrobial resistance profiles, virulence factors, plasmid content, serotypes and phylogenetic relationships. Methods: DNA was extracted and sequenced on the NovaSeq 6000 platform; the pangenome was assembled using the Roary tool; and the phylogenetic tree was constructed via IQ-TREE. Results and Discussion: For contextualization and comparison, 3493 Salmonella genomes of Brazilian origin from NCBI were analyzed. In our isolates, both strains carried the aac(6′)-Iaa_1 gene, while only Schwarzengrund harbored the qnrB19_1 gene and the Col440I_1 plasmid. Cerro presented the islands SPI-1, SPI-2, SPI-3, SPI-4, SPI-5 and SPI-9, while Schwarzengrund also possessed SPI-13 and SPI-14. Upon comparison with other Brazilian genomes, we observed that Cerro and Schwarzengrund represented only 0.40% and 2.03% of the national database, respectively. Furthermore, they revealed that Schwarzengrund presented higher levels of antimicrobial resistance, a finding supported by the higher frequency of plasmids in this serovar. Furthermore, national data corroborated our findings that SPI-13 and SPI-14 were absent in Cerro. A virulence analysis revealed distinct profiles: the cdtB and pltABC genes were present in the Schwarzengrund isolates, while the sseK and tldE1 family genes were exclusive to Cerro. The results indicated that the sequenced strains have pathogenic potential but exhibit low levels of antimicrobial resistance compared to national data. The greater diversity of SPIs in Schwarzengrund explains their prevalence and higher virulence potential. Conclusions: Finally, the serovars exhibit distinct virulence profiles, which results in different clinical outcomes. Full article

Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized with S. Typhimurium membrane vesicles (MVs). Silver nanoparticles (Ag NPs) were initially encapsulated within biodegradable poly(lactic-co-glycolic) nanoparticles (Ag-P NPs), which were then surface-modified with polyethyleneimine to form Ag-PP NPs. These were subsequently fused with S. Typhimurium MVs via a sonication method to generate Ag-PP@MV NPs. The resulting MV-coated nanoparticles displayed a similar protein profile to that of native MVs and exhibited antibacterial activity against intracellular S. Typhimurium. Notably, the enhanced cellular uptake of the MV-modified NPs contributed to their intracellular bactericidal efficacy. This study highlights MV modification as a promising strategy to improve NP delivery to macrophages for treating persistent intracellular infections. Full article

(1) Background: Understanding the distribution of Salmonella serovars in food, animals, and their environments is crucial for identifying infection sources and monitoring pathogen prevalence in the food chain. This study analysed Salmonella serovars in Poland from 2016 to 2020, focusing on their epidemiological significance. (2) Methods: Isolation of Salmonella was carried out following PN-EN ISO 6579 standards, and serotyping was performed using the White–Kauffmann–Le Minor scheme. A total of 7104 isolates were collected from food-producing animals, their environments, food of animal origin, feedingstuffs, and fertilisers. (3) Results: A total of 175 serovars were identified, with S. Enteritidis (n = 2905; 40.9%), S. Infantis (n = 1167; 16.4%), and S. Typhimurium (n = 360; 5.1%) being the most prevalent. Species-specific patterns were observed: S. Enteritidis dominated in chickens, ducks, and cattle; S. Kentucky in turkeys; S. Typhimurium in geese; and monophasic S. Typhimurium in pigs. S. Enteritidis and S. Infantis were most frequent in food of animal origin, especially broiler meat. In feedingstuffs, S. Agona was predominant, while fertilisers mostly contained S. Derby and S. Infantis. (4) Conclusions: The study highlights the source-dependent variety of Salmonella serovars and the importance of serotyping in tracing infection routes and preventing the spread of pathogens. Identifying the most common serovars supports the development of targeted preventive measures, including improved biosecurity, hygiene, and management practices to enhance food safety. Full article

Background/Objective: The present study focused on the analysis of the Spanish clone belonging to the successful 4,[5],12:i:- monophasic variant of Salmonella enterica serovar Typhimurium. Methods: All isolates of the clone recovered in a Spanish region from human clinical samples between 2008 and 2018 (N = 14) were investigated using microbiological approaches and genome sequence analysis. In addition, they were compared with isolates from the years 2000 to 2003 (N = 21), which were previously characterized but had not yet been sequenced. Results: Phylogenetic analyses indicate that all isolates are closely related (differing by 1 to 103 SNPs) but belong to two clades termed A and B. With few exceptions, clade A comprised isolates of the first period, also including two “older” control strains, LSP 389/97 and LSP 272/98. Clade B only contained isolates from the second period. Isolates from both periods were resistant to antibiotics and biocides, with almost all resistance genes located on large IncC plasmids, additionally carrying pSLT-derived virulence genes. The number of resistance genes was highly variable, resulting in a total of 22 ABR (antibiotic biocide resistance) profiles. The number of antibiotic resistance genes, but not that of biocide resistance genes, was considerably lower in isolates from the second than from the first period (with averages of 5.5 versus 9.6 genes). Importantly, IS26, which resides in multiple copies within these plasmids, appears to be playing a crucial role in the evolution of resistance, and it was also responsible for the monophasic phenotype, which was associated with four different deletions eliminating the fljAB region. Conclusions: the observed reduction in the number of antibiotic resistance genes could correlate with the loss of adaptive advantage originating from the ban on the use of antibiotics as feed additives implemented in the European Union since 2006, facilitated by the intrinsic instability of the IncC plasmids. Two consecutive IS26 transposition events, which can explain both the clonal relationship of the isolates and their variability, may account for the observed fljAB deletions. Full article

Salmonella colonization in the gastrointestinal tract of turkeys presents a risk to the safety of products derived from them. Lactobacillus-based probiotics and a plant-derived compound, trans-cinnamaldehyde, have previously been found to be effective in reducing multidrug-resistant Salmonella enterica subsp. enterica serovar Heidelberg (S. Heidelberg) in turkey poults. However, the effect of the challenge and the application of the treatments on the cecal metabolome has yet to be elucidated. Thus, the objective of the present study was to characterize alterations in the metabolic profiles of cecal contents collected from poults following S. Heidelberg challenge and treatment with Lactobacillus salivarius UMNPBX2 and L. ingluviei UMNPBX19 (LB), trans-cinnamaldehyde (TC), or a combination of both (CO) using untargeted gas chromatography–mass spectrometry (GC-MS). Poults in the challenged control (PC) group had the most distinct and convergent metabolome profiles, with the most pronounced disparity observed compared to the unchallenged control (NC), indicating the effect of the S. Heidelberg challenge. Perturbations in metabolites in the primary bile acid biosynthesis, pentose and glucuronate interconversions, and steroid biosynthesis were the most prominent. The greater abundance of metabolites, such as primary bile acids and sugars, in the PC group may be associated with S. Heidelberg colonization or potential shifts in microbiota. The treatments yielded varying effects on the metabolome profiles, with the TC and CO groups exhibiting the closest similarity, although TC was more similar to NC. The findings revealed alterations to ceca-associated metabolites, which are likely a response to the S. Heidelberg challenge and the application of the TC and LB treatments. Additional studies are needed to validate the possible causal relationship between the observed shifts. Gaining insight into the alterations to the metabolic microenvironment in the avian cecum will help elucidate the mechanisms by which they facilitate Salmonella persistence. Understanding these relationships can aid in designing more effective pre-harvest Salmonella control strategies and enhancing the efficacy of interventions within the flock. Full article

Antimicrobial use in food animals selects for antimicrobial-resistant (AMR) bacteria, which most commonly reach humans via the food chain. However, AMR bacteria can also escape the feedyard via agricultural runoff, manure used as crop fertilizer, and even dust. A study published in 2015 reported AMR genes in dust from cattle feedyards; however, one of the study’s major limitations was the failure to investigate gene presence in viable bacteria, or more importantly, viable bacteria of importance to human health. Our main objective was to investigate the presence and quantity of viable bacteria and antimicrobial-resistant (AMR) determinants in fugitive bioaerosols from cattle feedyards in the downwind environment. Six bioaerosol sampling campaigns were conducted at three commercial beef cattle feedyards to assess variability in viable bacteria and AMR determinants associated with geographic location, meteorological conditions, and season. Dust samples were collected using four different sampling methods, and spiral plated in triplicate on both non-selective and antibiotic-selective media. Colonies of total aerobic bacteria, Enterococcus spp., Salmonella enterica, and Escherichia coli were enumerated. Viable bacteria, including AMR bacteria, were identified in dust from cattle feedyards. Bacteria and antimicrobial resistance genes (ARGs via qPCR) were mainly found in downwind samples. Total suspended particles (TSPs) and impinger samples yielded the highest bacterial counts. Genes encoding beta-lactam resistance (bla_CMY-2 and bla_CTX-M) were detected while the most common ARG was tet(M). The predominant Salmonella serovar identified was Lubbock. Further research is needed to assess how far viable AMR bacteria can travel in the ambient environment downwind from cattle feedyards, to model potential public health risks. Full article