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11 pages, 1947 KiB  
Article
Exploring the Fermentation Profile, Bacterial Community, and Co-Occurrence Network of Big-Bale Leymus chinensis Silage Treated with/Without Lacticaseibacillus rhamnosus and Molasses
by Baiyila Wu, Xue Cao, Mingshan Fu, Yuxin Bao, Tiemei Wu, Kai Liu, Shubo Wen, Fenglin Gao, Haifeng Wang, Hua Mei and Yang Song
Agronomy 2025, 15(8), 1888; https://doi.org/10.3390/agronomy15081888 - 5 Aug 2025
Abstract
The purpose of this study was to investigate the effect of different additives on the microbial composition, fermentation quality, and bacterial community structure of big-bale Leymus chinensis silage. An experiment was set up with four treatment groups: a control (C) group, Lacticaseibacillus rhamnosus [...] Read more.
The purpose of this study was to investigate the effect of different additives on the microbial composition, fermentation quality, and bacterial community structure of big-bale Leymus chinensis silage. An experiment was set up with four treatment groups: a control (C) group, Lacticaseibacillus rhamnosus (L) group, molasses (M) group, and L. rhamnosus + molasses (LM) group, with three replications per group, and L. chinensis silages were fermented for 20 and 40 days. The lactic acid, acetic acid, 1,2-propanediol, and propionic acid contents increased, and pH, butyric acid, 1-propanol, and ethanol contents decreased in the L, M, and LM groups compared to the C group. In the LM group, the number of lactic acid bacteria was the highest, while the pH was the lowest. Enterobacter and Paucibacter were the main dominant genera in the C group. The addition of L. rhamnosus and molasses increased the relative abundance of Lactobacillus, Weissella, and Enterococcus. Lactobacillus abundance correlated positively (p < 0.01) with Lactococcus, Enterococcus, and Weissella and correlated negatively with Enterobacter and Paucibacter. Conversely, Enterobacter and Paucibacter showed a strong positive correlation (p < 0.01, R = 0.55) during fermentation. Lactobacillus, Enterococcus, and Weissella were positively associated (p < 0.01) with acetic and lactic acid levels, while Enterobacter abundance was correlated positively (p < 0.05, R = 0.43) with 1,2-propanediol content. In summary, the addition of both L. rhamnosus and molasses improved the fermentation quality and bacterial community structure of big-bale L. chinensis silage. In addition to inhibiting harmful microorganisms, this combination improved the fermentation products of big-bale L. chinensis silage through microbial regulation. Full article
(This article belongs to the Special Issue Innovative Solutions for Producing High-Quality Silage)
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24 pages, 3176 KiB  
Article
Influence of Seasonality and Pollution on the Presence of Antibiotic Resistance Genes and Potentially Pathogenic Bacteria in a Tropical Urban River
by Kenia Barrantes-Jiménez, Bradd Mendoza-Guido, Eric Morales-Mora, Luis Rivera-Montero, José Montiel-Mora, Luz Chacón-Jiménez, Keilor Rojas-Jiménez and María Arias-Andrés
Antibiotics 2025, 14(8), 798; https://doi.org/10.3390/antibiotics14080798 (registering DOI) - 5 Aug 2025
Abstract
Background/Objectives: This study examines how seasonality, pollution, and sample type (water and sediment) influence the presence and distribution of antibiotic resistance genes (ARGs), with a focus on antibiotic resistance genes (ARGs) located on plasmids (the complete set of plasmid-derived sequences, including ARGs) in [...] Read more.
Background/Objectives: This study examines how seasonality, pollution, and sample type (water and sediment) influence the presence and distribution of antibiotic resistance genes (ARGs), with a focus on antibiotic resistance genes (ARGs) located on plasmids (the complete set of plasmid-derived sequences, including ARGs) in a tropical urban river. Methods: Samples were collected from three sites along a pollution gradient in the Virilla River, Costa Rica, during three seasonal campaigns (wet 2021, dry 2022, and wet 2022). ARGs in water and sediment were quantified by qPCR, and metagenomic sequencing was applied to analyze chromosomal and plasmid-associated resistance profiles in sediments. Tobit and linear regression models, along with multivariate ordination, were used to assess spatial and seasonal trends. Results: During the wet season of 2021, the abundance of antibiotic resistance genes (ARGs) such as sul-1, intI-1, and tetA in water samples decreased significantly, likely due to dilution, while intI-1 and tetQ increased in sediments, suggesting particle-bound accumulation. In the wet season 2022, intI-1 remained low in water, qnrS increased, and sediments showed significant increases in tetQ, tetA, and qnrS, along with decreases in sul-1 and sul-2. Metagenomic analysis revealed spatial differences in plasmid-associated ARGs, with the highest abundance at the most polluted site (Site 3). Bacterial taxa also showed spatial differences, with greater plasmidome diversity and a higher representation of potential pathogens in the most contaminated site. Conclusions: Seasonality and pollution gradients jointly shape ARG dynamics in this tropical river. Plasmid-mediated resistance responds rapidly to environmental change and is enriched at polluted sites, while sediments serve as long-term reservoirs. These findings support the use of plasmid-based monitoring for antimicrobial resistance surveillance in aquatic systems. Full article
(This article belongs to the Special Issue Origins and Evolution of Antibiotic Resistance in the Environment)
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23 pages, 4178 KiB  
Article
Taxonomic Biomarkers of Gut Microbiota with Potential Clinical Utility in Mexican Adults with Obesity and Depressive and Anxiety Symptoms
by María Alejandra Samudio-Cruz, Daniel Cerqueda-García, Elizabeth Cabrera-Ruiz, Alexandra Luna-Angulo, Samuel Canizales-Quinteros, Carlos Landa-Solis, Gabriela Angélica Martínez-Nava, Paul Carrillo-Mora, Edgar Rangel-López, Juan Ríos-Martínez, Blanca López-Contreras, Jesús Fernando Valencia-León and Laura Sánchez-Chapul
Microorganisms 2025, 13(8), 1828; https://doi.org/10.3390/microorganisms13081828 - 5 Aug 2025
Abstract
While the gut microbiota of obese children in Mexico has been studied, its relationship with depressive and anxiety symptoms in obese adults remains unexplored. The aim of this study was to describe the gut microbiota profile of Mexican adults with obesity and its [...] Read more.
While the gut microbiota of obese children in Mexico has been studied, its relationship with depressive and anxiety symptoms in obese adults remains unexplored. The aim of this study was to describe the gut microbiota profile of Mexican adults with obesity and its association with depression and anxiety. We sequenced the V3-V4 region of the 16S rRNA gene from stool samples of obese adults categorized into four groups: control (OCG), with depressive symptoms (OD), with anxiety symptoms (OAx), or with both (ODAx). Alpha diversity was assessed using t-tests, beta diversity was assessed with PERMANOVA, and taxonomic differences was assessed with LEfSe. Associations between bacterial genera and clinical variables were analyzed using the Maaslin2 library. Bacteroidota was the most prevalent phylum, and Prevotella was the dominant enterotype across all groups. Although overall diversity did not differ significantly, 30 distinct taxonomic biomarkers were identified among groups as follows: 4 in OCG (Firmicutes), 5 in OD (Firmicutes, Bacteroidota), 13 in OAx (Firmicutes, Bacteroidetes, Fusobacteroidota, Proteobacteria), and 8 in ODAx (Firmicutes). This is the first study to identify distinct gut microbiota profiles in obese Mexican adults with depressive and anxiety symptoms. These findings suggest important microbial biomarkers for improving the diagnosis and treatment of mental health conditions in obesity. Full article
(This article belongs to the Special Issue Gut Microbiota: Influences and Impacts on Human Health)
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12 pages, 388 KiB  
Article
Evolution of Respiratory Pathogens and Antimicrobial Resistance over the COVID-19 Timeline: A Study of Hospitalized and Ambulatory Patient Populations
by Luigi Regenburgh De La Motte, Loredana Deflorio, Erika Stefano, Matteo Covi, Angela Uslenghi, Carmen Sommese and Lorenzo Drago
Antibiotics 2025, 14(8), 796; https://doi.org/10.3390/antibiotics14080796 (registering DOI) - 5 Aug 2025
Abstract
Background: The COVID-19 pandemic has profoundly altered the clinical and microbiological landscape of respiratory tract infections (RTIs), potentially reshaping pathogen distribution and antimicrobial resistance (AMR) profiles across care settings. Objectives: The objective of this study was to assess temporal trends in respiratory bacterial [...] Read more.
Background: The COVID-19 pandemic has profoundly altered the clinical and microbiological landscape of respiratory tract infections (RTIs), potentially reshaping pathogen distribution and antimicrobial resistance (AMR) profiles across care settings. Objectives: The objective of this study was to assess temporal trends in respiratory bacterial pathogens, antimicrobial resistance, and polymicrobial infections across three pandemic phases—pre-COVID (2018–2019), COVID (2020–2022), and post-COVID (2022–2024)—in hospitalized and ambulatory patients. Methods: We retrospectively analyzed 1827 respiratory bacterial isolates (hospitalized patients, n = 1032; ambulatory patients, n = 795) collected at a tertiary care center in Northern Italy. Data were stratified by care setting, anatomical site, and pandemic phase. Species identification and susceptibility testing followed EUCAST guidelines. Statistical analysis included chi-square and Fisher’s exact tests. Results: In hospitalized patients, a significant increase in Pseudomonas aeruginosa (from 45.5% pre-COVID to 58.6% post-COVID, p < 0.0001) and Acinetobacter baumannii (from 1.2% to 11.1% during COVID, p < 0.0001) was observed, with 100% extensively drug-resistant (XDR) rates for A. baumannii during the pandemic. Conversely, Staphylococcus aureus significantly declined from 23.6% pre-COVID to 13.7% post-COVID (p = 0.0012). In ambulatory patients, polymicrobial infections peaked at 41.2% during COVID, frequently involving co-isolation of Candida spp. Notably, resistance to benzylpenicillin in Streptococcus pneumoniae reached 80% (4/5 isolates) in hospitalized patients during COVID, and carbapenem-resistant P. aeruginosa (CRPA) significantly increased post-pandemic in ambulatory patients (0% pre-COVID vs. 23.5% post-COVID, p = 0.0014). Conclusions: The pandemic markedly shifted respiratory pathogen dynamics and resistance profiles, with distinct trends observed in hospital and community settings. Persistent resistance phenotypes and frequent polymicrobial infections, particularly involving Candida spp. in outpatients, underscore the need for targeted surveillance and antimicrobial stewardship strategies. Full article
(This article belongs to the Section Antibiotic Therapy in Infectious Diseases)
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18 pages, 903 KiB  
Article
Effect of Allyl-Isothiocyanate Release from Black Mustard (Brassica nigra) Seeds During Refrigerated Storage to Preserve Fresh Tench (Tinca tinca) Fillets
by María José Rodríguez Gómez, María Alejo Martínez, Raquel Manzano Durán, Daniel Martín-Vertedor and Patricia Calvo Magro
Fishes 2025, 10(8), 381; https://doi.org/10.3390/fishes10080381 - 5 Aug 2025
Abstract
The aim of this study was to prevent the development of microorganisms in the refrigerated storage of tench by releasing allyl isothiocyanate (AITC) produced by black mustard seeds. Tench reared in an aquaculture centre were sacrificed and the fillets were separated. Different amounts [...] Read more.
The aim of this study was to prevent the development of microorganisms in the refrigerated storage of tench by releasing allyl isothiocyanate (AITC) produced by black mustard seeds. Tench reared in an aquaculture centre were sacrificed and the fillets were separated. Different amounts of defatted mustard seed (300, 400 and 500 mg) were added to hermetic polypropylene trays. Microbiological, sensory, and gas chromatography with MS detection analysis were done. AITC release increased progressively until the third day of storage, significantly delaying the development of microorganisms in samples with higher mustard seed content. The tasting panel detected positive aromas at the beginning of the study, but these decreased and negative aromas appeared. The mustard seed treatment resulted in a higher positive aroma at the end of the storage, reducing rotting and ammonia odours. A total of 31 volatile compounds were detected and grouped into hydrocarbon, alcohol, benzenoid, isothiocyanate, ketone, acetate, aldehyde, and others. Butylated hydroxytoluene, an indicator of bacterial contamination, was the major aromatic compound found during storage. The release of AITC resulted in fewer organic compounds with negative aromas appearing during storage. PCA analysis allowed us to classify the assays during storage according to their volatile profiles, confirming the differences observed between treatments. Thus, adding mustard seed to fish packaging could be a viable alternative to extending the product’s shelf life and ensuring food safety. Full article
(This article belongs to the Section Processing and Comprehensive Utilization of Fishery Products)
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37 pages, 5366 KiB  
Article
Oral Microbiota Composition and Its Association with Gastrointestinal and Developmental Abnormalities in Children with Autism Spectrum Disorder
by Zuzanna Lewandowska-Pietruszka, Magdalena Figlerowicz and Katarzyna Mazur-Melewska
Microorganisms 2025, 13(8), 1822; https://doi.org/10.3390/microorganisms13081822 - 4 Aug 2025
Abstract
Autism Spectrum Disorder (ASD) is frequently accompanied by gastrointestinal disturbances, dietary selectivity, and altered stress responses, with growing evidence pointing to gut–brain axis involvement. While intestinal microbiota has been extensively studied, the role of the oral microbiota remains underexplored. This study investigates the [...] Read more.
Autism Spectrum Disorder (ASD) is frequently accompanied by gastrointestinal disturbances, dietary selectivity, and altered stress responses, with growing evidence pointing to gut–brain axis involvement. While intestinal microbiota has been extensively studied, the role of the oral microbiota remains underexplored. This study investigates the associations between oral microbiota composition and behavioral, gastrointestinal, dietary, and neuroendocrine parameters in children with ASD. A total of 45 children aged 2–18 years comprised the study group. Data collection included oral swabs for 16S rRNA gene sequencing, salivary cortisol sampling, dietary records, and standardized behavioral assessments using the Vineland Adaptive Behavior Scale. A total of 363 microbial species across 11 phyla were identified. Significant correlations were observed between specific bacterial taxa and functional gastrointestinal disorders (FGIDs), dietary patterns, salivary cortisol rhythms, and functioning. Children with FGIDs, food selectivity, or macronutrient imbalances exhibited enriched pro-inflammatory taxa (e.g., Selenomonas, Megasphaera), whereas those with typical cortisol secretion or higher adaptive functioning showed greater microbial diversity and abundance of health-associated genera (e.g., Bifidobacterium dentium). These findings suggest that oral microbiota profiles may reflect systemic physiological and neurobehavioral traits in children with ASD. Further longitudinal studies are needed to clarify causal relationships and support the development of microbiota-targeted interventions. Full article
(This article belongs to the Special Issue Focus on Pediatric Infectious Diseases)
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15 pages, 647 KiB  
Article
Effects of Burdock Addition and Different Starters on the Quality and Flavor Improvement of Duck Sausages
by Li Cui, Xuan Zhao, Xingye Song, Wenjing Zhou, Tao Wang, Wuyang Huang and Yuxing Guo
Biology 2025, 14(8), 996; https://doi.org/10.3390/biology14080996 (registering DOI) - 4 Aug 2025
Abstract
Burdock (Arctium lappa L.) is a medicinal and edible homologous plant whose roots contain many bioactive substances such as polysaccharides and phenolics. This study explored the integration of burdock powder and lactic acid bacteria fermentation to enhance the nutritional quality, sensory attributes, [...] Read more.
Burdock (Arctium lappa L.) is a medicinal and edible homologous plant whose roots contain many bioactive substances such as polysaccharides and phenolics. This study explored the integration of burdock powder and lactic acid bacteria fermentation to enhance the nutritional quality, sensory attributes, and flavor profiles of duck sausages. Three bacterial strains, Lacticaseibacillus casei, L. helveticus, and L. plantarum, were selected based on sensory analysis, and their effects on sausage properties were evaluated through combined fermentation trials. The results demonstrated that duck sausages fermented with L. plantarum and L. helveticus and supplemented with 3% burdock powder (PHB group) exhibited > 1.5-fold higher antioxidant activity (ABTS at 85.2 μmol trolox/g and DPPH at 92.7 μmol trolox/g, respectively; p < 0.05) and 15% increase in total phenolic content (8.24 mg gallic acid/g) compared to non-fermented counterparts. The PHB formulation also enhanced color stability (lightness, redness, yellowness), textural characteristics (hardness, springiness, cohesiveness), and sensory acceptability. Volatile compound analysis revealed a reduction in off-odor aldehydes (hexanal, (E)-2-octenal, (E)-2-decenal, and (E,E)-2,4-decadienal) and increased production of desirable aromatic compounds like tetramethyl-pyrazine. These findings highlight the potential of combining lactic acid bacteria fermentation with burdock powder to develop functional duck sausages with improved nutritional and sensory properties. Full article
(This article belongs to the Special Issue Nutraceutical and Bioactive Compounds in Foods)
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28 pages, 2282 KiB  
Article
From Hue to Health: Exploring the Therapeutic Potential of Plant-Pigment-Enriched Extracts
by Azza SalahEldin El-Demerdash, Amira E. Sehim, Abeer Altamimi, Hanan Henidi, Yasmin Mahran and Ghada E. Dawwam
Microorganisms 2025, 13(8), 1818; https://doi.org/10.3390/microorganisms13081818 - 4 Aug 2025
Abstract
The escalating global challenges of antimicrobial resistance (AMR) and cancer necessitate innovative therapeutic solutions from natural sources. This study investigated the multifaceted therapeutic potential of pigment-enriched plant extracts. We screened diverse plant extracts for antimicrobial and antibiofilm activity against multidrug-resistant bacteria and fungi. [...] Read more.
The escalating global challenges of antimicrobial resistance (AMR) and cancer necessitate innovative therapeutic solutions from natural sources. This study investigated the multifaceted therapeutic potential of pigment-enriched plant extracts. We screened diverse plant extracts for antimicrobial and antibiofilm activity against multidrug-resistant bacteria and fungi. Hibiscus sabdariffa emerged as the most promising, demonstrating potent broad-spectrum antimicrobial and significant antibiofilm activity. Sub-inhibitory concentrations of H. sabdariffa robustly downregulated essential bacterial virulence genes and suppressed aflatoxin gene expression. Comprehensive chemical profiling via HPLC identified major anthocyanin glucosides, while GC-MS revealed diverse non-pigment bioactive compounds, including fatty acids and alcohols. Molecular docking suggested favorable interactions of key identified compounds (Cyanidin-3-O-glucoside and 1-Deoxy-d-arabitol) with E. coli outer membrane protein A (OmpA), indicating potential antiadhesive and antimicrobial mechanisms. Furthermore, H. sabdariffa exhibited selective cytotoxicity against MCF-7 breast cancer cells. These findings establish H. sabdariffa pigment-enriched extract as a highly promising, multi-functional source of novel therapeutics, highlighting its potential for simultaneously addressing drug resistance and cancer challenges through an integrated chemical, biological, and computational approach. Full article
(This article belongs to the Special Issue Advanced Research on Antimicrobial Activity of Natural Products)
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22 pages, 2692 KiB  
Article
Differences in the Profile of Aromatic Metabolites in the Corresponding Blood Serum and Cerebrospinal Fluid Samples of Patients with Secondary Bacterial Meningitis
by Alisa K. Pautova, Peter A. Meinarovich, Vladislav E. Zakharchenko, Pavel D. Sobolev, Natalia A. Burnakova and Natalia V. Beloborodova
Metabolites 2025, 15(8), 527; https://doi.org/10.3390/metabo15080527 (registering DOI) - 3 Aug 2025
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Abstract
Background: Secondary (nosocomial) bacterial meningitis remains a serious problem in patients with severe brain damage. The aim of this study was to assess the differences in the aromatic metabolites of tryptophan, phenylalanine, and tyrosine, in serum and cerebrospinal fluid (CSF) samples collected simultaneously [...] Read more.
Background: Secondary (nosocomial) bacterial meningitis remains a serious problem in patients with severe brain damage. The aim of this study was to assess the differences in the aromatic metabolites of tryptophan, phenylalanine, and tyrosine, in serum and cerebrospinal fluid (CSF) samples collected simultaneously from patients with long-term sequelae of severe brain damage with suspected secondary bacterial meningitis. Methods: Group I included 16 paired serum and CSF samples from patients (N = 11) without secondary bacterial meningitis; group II included 13 paired serum and CSF samples from patients (N = 4) with secondary bacterial meningitis. Results: The median concentrations of serum 5-hydroxyindole-3-acetic, CSF 4-hydroxyphenyllactic (p-HPhLA), CSF 4-hydroxyphenylacetic, CSF phenyllactic, and indole-3-lactic acids in serum and CSF were statistically higher in group II compared to group I (p-value ≤ 0.03), while 4-hydroxyphenylpropionic and indole-3-acetic in serum were lower in group II compared to group I (p-value = 0.04). In group I, p-HPhLA serum concentrations were greater than or equal to its CSF concentrations in 14 paired samples; in group II, p-HPhLA concentrations in serum were lower than in CSF in all paired samples. Conclusions: The obtained results demonstrate the differences in the profile of aromatic metabolites in serum and CSF and may confirm the hypothesis of the p-HPhLA microbial origin in the CSF of patients with secondary bacterial meningitis. Full article
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21 pages, 3431 KiB  
Article
Synthesis and Antibacterial Evaluation of an Indole Triazole Conjugate with In Silico Evidence of Allosteric Binding to Penicillin-Binding Protein 2a
by Vidyasrilekha Sanapalli, Bharat Kumar Reddy Sanapalli and Afzal Azam Mohammed
Pharmaceutics 2025, 17(8), 1013; https://doi.org/10.3390/pharmaceutics17081013 - 3 Aug 2025
Viewed by 79
Abstract
Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial [...] Read more.
Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial cell wall biosynthesis. Objectives: The objective was to design and characterize a novel small-molecule inhibitor targeting PBP2a as a strategy to combat MRSA. Methods: We synthesized a new indole triazole conjugate (ITC) using eco-friendly and click chemistry approaches. In vitro antibacterial tests were performed against a panel of strains to evaluate the ITC antibacterial potential. Further, a series of in silico evaluations like molecular docking, MD simulations, free energy landscape (FEL), and principal component analysis (PCA) using the crystal structure of PBP2a (PDB ID: 4CJN), in order to predict the mechanism of action, binding mode, structural stability, and energetic profile of the 4CJN-ITC complex. Results: The compound ITC exhibited noteworthy antibacterial activity, which effectively inhibited the selected strains. Binding score and energy calculations demonstrated high affinity of ITC for the allosteric site of PBP2a and significant interactions responsible for complex stability during MD simulations. Further, FEL and PCA provided insights into the conformational behavior of ITC. These results gave the structural clues for the inhibitory action of ITC on the PBP2a. Conclusions: The integrated in vitro and in silico studies corroborate the potential of ITC as a promising developmental lead targeting PBP2a in MRSA. This study demonstrates the potential usage of rational drug design approaches in addressing therapeutic needs related to ABR. Full article
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20 pages, 4612 KiB  
Article
Effect of a Gluten-Free Diet on the Intestinal Microbiota of Women with Celiac Disease
by M. Mar Morcillo Serrano, Paloma Reche-Sainz, Daniel González-Reguero, Marina Robas-Mora, Rocío de la Iglesia, Natalia Úbeda, Elena Alonso-Aperte, Javier Arranz-Herrero and Pedro A. Jiménez-Gómez
Antibiotics 2025, 14(8), 785; https://doi.org/10.3390/antibiotics14080785 (registering DOI) - 2 Aug 2025
Viewed by 173
Abstract
Background/Objectives: Celiac disease (CD) is an autoimmune disorder characterized by small intestinal enteropathy triggered by gluten ingestion, often associated with gut dysbiosis. The most effective treatment is strict adherence to a gluten-free diet (GFD), which alleviates symptoms. This study uniquely integrates taxonomic, [...] Read more.
Background/Objectives: Celiac disease (CD) is an autoimmune disorder characterized by small intestinal enteropathy triggered by gluten ingestion, often associated with gut dysbiosis. The most effective treatment is strict adherence to a gluten-free diet (GFD), which alleviates symptoms. This study uniquely integrates taxonomic, functional, and resistance profiling to evaluate the gut microbiota of women with CD on a GFD. Methods: To evaluate the long-term impact of a GFD, this study analyzed the gut microbiota of 10 women with CD on a GFD for over a year compared to 10 healthy controls with unrestricted diets. Taxonomic diversity (16S rRNA gene sequencing and the analysis of α and β-diversity), metabolic functionality (Biolog EcoPlates®), and antibiotic resistance profiles (Cenoantibiogram) were assessed. Results: Metagenomic analysis revealed no significant differences in taxonomic diversity but highlighted variations in the abundance of specific bacterial genera. Women with CD showed increased proportions of Bacteroides, Streptococcus, and Clostridium, associated with inflammation, but also elevated levels of beneficial genera such as Roseburia, Oxalobacter, and Paraprevotella. Despite no significant differences in metabolic diversity, higher minimum inhibitory concentrations (MICs) in women in the healthy control group suggest that dietary substrates in unrestricted diets may promote the proliferation of fast-growing bacteria capable of rapidly developing and disseminating antibiotic resistance mechanisms. Conclusions: These findings indicate that prolonged adherence to a GFD in CD supports remission of gut dysbiosis, enhances microbiota functionality, and may reduce the risk of antibiotic resistance, emphasizing the importance of dietary management in CD. Full article
(This article belongs to the Special Issue Antibiotic Resistance: A One-Health Approach, 2nd Edition)
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23 pages, 4116 KiB  
Article
Taxonomic and Functional Profiling of Bacterial Communities in Leather Biodegradation: Insights into Metabolic Pathways and Diversity
by Manuela Bonilla-Espadas, Marcelo Bertazzo, Irene Lifante-Martinez, Mónica Camacho, Elena Orgilés-Calpena, Francisca Arán-Aís and María-José Bonete
Bacteria 2025, 4(3), 37; https://doi.org/10.3390/bacteria4030037 - 1 Aug 2025
Viewed by 87
Abstract
Leather biodegradation is a complex microbial process with increasing relevance for sustainable waste management. In this study, we investigated bacterial communities responsible for the degradation of leather treated with different tanning agents (chrome, Zeolite, Biole®) using high-throughput 16S rRNA gene sequencing [...] Read more.
Leather biodegradation is a complex microbial process with increasing relevance for sustainable waste management. In this study, we investigated bacterial communities responsible for the degradation of leather treated with different tanning agents (chrome, Zeolite, Biole®) using high-throughput 16S rRNA gene sequencing and metatranscriptomic analysis. Proteobacteria, Bacteroidetes, and Patescibacteria emerged as the dominant phyla, while genera such as Acinetobacter, Pseudomonas, and Sphingopyxis were identified as key contributors to enzymatic activity and potential metal resistance. A total of 1302 enzymes were expressed across all the conditions, including 46 proteases, with endopeptidase La, endopeptidase Clp, and methionyl aminopeptidase being the most abundant. Collagen samples exhibited the highest functional diversity and total enzyme expression, whereas chrome-treated samples showed elevated protease activity, indicating selective pressure from heavy metals. Differential enzyme expression patterns were linked to both the microbial identity and tanning chemistry, revealing genus- and treatment-specific enzymatic signatures. These findings deepen our understanding of how tanning agents modulate the microbial structure and function and identify proteases with potential applications in the bioremediation and eco-innovation of leather waste processing. Full article
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23 pages, 3211 KiB  
Article
Investigation of Bacterial Species and Their Antimicrobial Drug Resistance Profile in Feline Urinary Tract Infection in Thailand
by Kankanit Lapcharoen, Chunyaput Bumrungpun, Wiyada Chumpol, Kamonwan Lunha, Suganya Yongkiettrakul, Porntippa Lekcharoensuk and Chantima Pruksakorn
Animals 2025, 15(15), 2235; https://doi.org/10.3390/ani15152235 - 30 Jul 2025
Viewed by 282
Abstract
Feline urinary tract infections (UTIs) present a common challenge in veterinary practice, underscoring the importance of understanding local bacterial pathogens and antimicrobial resistance (AMR). This study determined bacterial prevalence and antimicrobial susceptibility in cats at Kasetsart University’s Veterinary Teaching Hospital in Bangkok, Thailand. [...] Read more.
Feline urinary tract infections (UTIs) present a common challenge in veterinary practice, underscoring the importance of understanding local bacterial pathogens and antimicrobial resistance (AMR). This study determined bacterial prevalence and antimicrobial susceptibility in cats at Kasetsart University’s Veterinary Teaching Hospital in Bangkok, Thailand. Of the 543 cystocentesis urine samples collected from 428 cats, 115 (21.2%) tested positive for bacterial cultures, leading to a diagnosis of UTIs in 95 cats (22.2%). The most prevalent isolates included Escherichia coli (24.8%), Staphylococcus species (19.2%), Proteus mirabilis (13.6%), Pseudomonas aeruginosa (12.0%), and Enterococcus species (12.0%). Staphylococcus felis (8.8%) and Staphylococcus pseudintermedius (5.6%) were the predominant Staphylococcus species. Rare pathogens such as Corynebacterium urealyticum and Lactococcus garvieae were also identified. Antimicrobial testing revealed alarming resistance, with 69.2% of isolates exhibiting multidrug resistance (MDR). Escherichia coli and Proteus mirabilis showed high resistance to amoxicillin/clavulanic acid (AMC) (45.2–70.6%) and sulfamethoxazole/trimethoprim (SXT) (51.6–52.9%). Enterococcus faecium exhibited 85.7% resistance to AMC. Methicillin resistance was identified in 41.7% of Staphylococcus isolates, particularly high in Staphylococcus epidermidis (75.0%) and Staphylococcus pseudintermedius (71.4%). High fluoroquinolone resistance among MDR isolates further exacerbates AMR concerns. These results indicate that MDR Gram-negative, Staphylococcus, and Enterococcus species complicate the empirical treatment of feline UTIs, highlighting significant implications for AMR in veterinary practice. Full article
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15 pages, 752 KiB  
Article
Enhanced Anti-Inflammatory Effects of Rosemary (Salvia rosmarinus) Extracts Modified with Pseudomonas shirazensis Nanoparticles
by Enrique Gutierrez-Albanchez, Elena Fuente-González, Svitlana Plokhovska, Francisco Javier Gutierrez-Mañero and Beatriz Ramos-Solano
Antioxidants 2025, 14(8), 931; https://doi.org/10.3390/antiox14080931 - 29 Jul 2025
Viewed by 250
Abstract
Rosemary (Salvia rosmarinus) is renowned for its antioxidant, anti-inflammatory, and antihyperglycemic properties, largely attributed to its rich phytochemical profile. This study evaluates the potential of metabolites from Pseudomonas shirazensis NFV3, formulated in silver nanoparticles (AgNPs), to enhance the bioactivity of rosemary [...] Read more.
Rosemary (Salvia rosmarinus) is renowned for its antioxidant, anti-inflammatory, and antihyperglycemic properties, largely attributed to its rich phytochemical profile. This study evaluates the potential of metabolites from Pseudomonas shirazensis NFV3, formulated in silver nanoparticles (AgNPs), to enhance the bioactivity of rosemary extracts in postharvest applications. Rosemary stems were treated with AgNPs coated with bacterial metabolites (NP), bacterial cells, or metabolites (LM), and the extracts’ phytochemical composition and bioactivities were assessed. HPLC and HPLC–MS analyses revealed that the NP treatment induced significant metabolic remodeling, particularly upregulating rosmarinic acid and selected triterpenes (ursolic and betulinic acids), while reducing carnosic acid levels. NP-treated extracts exhibited significantly enhanced inhibition of cyclooxygenase (COX-1 and COX-2), indicating improved anti-inflammatory potential. The α-glucosidase inhibition and antioxidant activity (DPPH assay) of the extracts were not substantially altered, suggesting the selective enhancement of pharmacological functions. These findings demonstrate that nanoparticle-based elicitation selectively remodels secondary metabolism in rosemary, improving extract quality and bioactivity. This strategy offers a novel, sustainable tool for optimizing plant-based therapeutics in the phytopharmaceutical industry. Full article
(This article belongs to the Special Issue Applications of Antioxidant Nanoparticles, 2nd Edition)
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Article
Streamlining Bacillus Strain Selection Against Listeria monocytogenes Using a Fluorescence-Based Infection Assay Integrated into a Multi-Tiered Validation Pipeline
by Blanca Lorente-Torres, Pablo Castañera, Helena Á. Ferrero, Sergio Fernández-Martínez, Suleiman Adejoh Ocholi, Jesús Llano-Verdeja, Farzaneh Javadimarand, Yaiza Carnicero-Mayo, Amanda Herrero-González, Alba Puente-Sanz, Irene Sainz Machín, Isabel Karola Voigt, Silvia Guerrero Villanueva, Álvaro López García, Eva Martín Gómez, James C. Ogbonna, José M. Gonzalo-Orden, Jesús F. Aparicio, Luis M. Mateos, Álvaro Mourenza and Michal Letekadd Show full author list remove Hide full author list
Antibiotics 2025, 14(8), 765; https://doi.org/10.3390/antibiotics14080765 - 29 Jul 2025
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Abstract
Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential [...] Read more.
Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential against L. monocytogenes. Methods: A total of 26 Bacillus isolates were screened for antimicrobial activity, gastrointestinal resilience, and host cell adhesion. A fluorescence-based infection assay using mCherry-expressing HCT 116 cells was used to assess cytoprotection against L. monocytogenes NCTC 7973. Eight strains significantly improved host cell viability and were validated by quantification of intracellular CFU. Two top candidates were tested in a murine model of listeriosis. The genome of the lead strain was sequenced to evaluate safety and biosynthetic potential. Results: B. subtilis CECT 8266 completely inhibited intracellular replication of L. monocytogenes in HCT 116 cells, reducing bacterial recovery to undetectable levels. In vivo, it decreased splenic bacterial burden by approximately 6-fold. Genomic analysis revealed eight bacteriocin biosynthetic clusters and silent antibiotic resistance genes within predicted genomic islands, as determined by CARD and Alien Hunter analysis. The strain also demonstrated bile and acid tolerance, as well as strong adhesion to epithelial cells. Conclusions: The proposed pipeline enables efficient identification of probiotic Bacillus strains with intracellular protective activity. B. subtilis CECT 8266 is a promising candidate for translational applications in food safety or health due to its efficacy, resilience, and safety profile. Full article
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