Search Results (558)

Lactic acid fermentation (LAF) is a promising low-tech approach for the stabilization and hygienization of human feces from dry toilets. However, practical implementation remains limited due to a lack of application-relevant knowledge. This study systematically evaluated the influence of a series of practical process parameters on the performance of LAF under standardized laboratory conditions. Feces obtained from different types of dry toilets were physicochemically characterized and subsequently fermented under varying process conditions, using pH and lactic acid production as key indicators of fermentation performance. The results indicate that LAF is feasible across a broad range of process conditions, including temperatures between 8 and 30 °C, and is largely independent of carbon source type, air intrusion, and extended storage periods (>1 year), provided that a sufficient carbon supply is ensured. The investigated parameters exhibited varying degrees of influence on process performance, with carbon source dosage (≥10 w/w-% sugar beet molasses equivalent) and feces type emerging as the most influential factors. While ferrous iron addition (≤5 w/w-%) enhanced pH reduction, biochar, bentonite, and rock flour (≤10 w/w-%) showed negligible effects. Maximum lactic acid production was limited to ≤4.5 w/w-%, irrespective of carbon source dosage, resulting in minimum pH values ranging from 4.1 to 5.2. These values varied primarily with fecal type, suggesting a strong influence of intrinsic buffering capacity. Under conditions supporting stable LAF, i.e., rapid acidification followed by sustained low pH, E. coli was consistently reduced below the detection limit in the investigated samples. Overall, the findings suggest that LAF is a comparatively robust treatment approach and highlight operational parameters that are likely to be important for its practical implementation as a sanitation technology. Full article

Extended-spectrum β-lactamase-producing Klebsiella pneumoniae (ESBL-KP) is an important antimicrobial-resistant pathogen, and wastewater may serve as a reservoir for its persistence and dissemination. This study investigated the virulence-associated genes, biofilm-forming capacity and genomic relatedness of ESBL-KP isolates recovered from wastewater and livestock farm environments in southeastern North Carolina. A cross-sectional study was conducted between May and September 2025 at two wastewater treatment plants (WWTPs) and two livestock farms. ESBL-KP isolates recovered from wastewater, animal feces, and water samples were characterized using PCR, whole-genome sequencing and crystal violet biofilm assays. Genomic relatedness was assessed using phylogenomic analysis. Data were analyzed using descriptive statistics and Fisher’s exact test. ESBL-KP was detected in 15.4% (n = 69/449) of samples, with the highest prevalence observed in WWTPs (75.4%, n = 52) followed by poultry farms (21.7%, n = 15). The most frequent virulence genes were mrkD (30/69), entB (26/69), K2 (21/69), and rmpA (21/69). Significant variation in gene distribution by sample type was observed for mrkD (p = 0.0013) and entB (p = 0.0011). Biofilm formation varied by sample type, with strong biofilm predominating in influent (n = 20) and sludge (n = 8), although no significant differences were detected across sample types (p = 0.357). Phylogenetic analysis revealed that one poultry farm isolate was clonally related to wastewater isolates, differing by 1–3 single nucleotide polymorphisms (SNPs) and sharing the virulence genes mrkA, iutA, and fimH. Overall, environmental ESBL-KP isolates exhibited widespread virulence potential and robust biofilm-forming capacity, while phylogenetic evidence demonstrated clonal relatedness between poultry farm and wastewater isolates and sharing mrkA, iutA, and fimH virulence genes. These findings highlight wastewater and agricultural systems as genetically related reservoirs for clinically relevant ESBL-KP strains and underscore the need for strengthened One Health-based surveillance to monitor and mitigate their environmental dissemination. Full article

With the growing interest in the microbiome, short-chain fatty acids (SCFAs) have emerged as key metabolites due to their critical roles in host physiology, including immune regulation, energy homeostasis, and inflammatory control. As a result, the accurate quantification of SCFAs in various biological samples has become increasingly important. However, reliable and standardized methods for measuring SCFAs across different sample types remain underdeveloped, highlighting the need for methodological refinement. To address this need, we optimized two analytical methods, headspace GC-MS and GC-MS/MS, for SCFA quantification. These techniques were applied to a range of biological matrices, including pure microbial cultures, low-abundance animal liver, animal feces, and standardized simulated human fecal samples. The headspace GC-MS approach enables direct analysis with minimal sample preparation, thereby enhancing throughput and ease of use. In contrast, the GC-MS/MS method, involving methanol extraction, alkaline treatment, and derivatization with MTBSTFA, offers superior sensitivity and precision, making it particularly suitable for small-volume and low-abundance samples. Together, these optimized protocols provide robust, sensitive platforms for profiling SCFAs across diverse biological matrices, facilitating a deeper understanding of microbiome–host interactions and supporting future translational applications. Full article

A novel Bacillus velezensis strain DY201, isolated from broiler feces, was characterized to assess its probiotic potential as an antibiotic alternative in poultry production. The strain demonstrated robust environmental tolerance with optimal growth at 42 °C and 51.32% survival following sequential exposure to simulated gastric and intestinal fluids. DY201 exhibited broad-spectrum antimicrobial activity against enterotoxigenic Escherichia coli K88, Staphylococcus aureus, Salmonella pullorum, and Clostridium perfringens, with activity remaining stable across pH 5.0–8.0 and retaining over 92.65% efficacy after 85 °C treatment. Scanning electron microscopy revealed metabolite-induced membrane perforation in target pathogens. Although whole-genome sequencing identified 14 biosynthetic gene clusters for lipopeptides including surfactin and fengycin, integrated proteomic and metabolomic analyses detected small-molecule metabolites—Withaferin A, 2′-hydroxy-2-methoxychalcone, and platycodigenin—as the primary antimicrobial effectors. In a preliminary broiler trial, dietary DY201 supplementation significantly increased the relative abundance of Bacillus in the ileum from 0.30% to 10.30% (p = 0.0434) and in the jejunum from 0.77% to 5.56% (p = 0.0453), enriched the generally beneficial genus Lactobacillus in the jejunum from 73.05% to 80.11% (p = 0.0323), and reduced Candidatus Arthromitus in the ileum from 13.38% to 0.59% (p = 0.0105). These findings support B. velezensis DY201 as a promising probiotic candidate for intestinal microbiota modulation in broilers, although functional intestinal health benefits require further validation through growth performance, barrier function, immune response, and pathogen challenge studies. Full article

Background: Overweight and obesity are chronic diseases that result from complex interactions including genetics, environment, eating behaviors, and limited access to a healthy diet. Amaranth protein (AmProt) has several health benefits, but no studies have examined its effects on the modulation of children’s gut microbiota. The work aimed to analyze serum levels and changes in gut microbiota in children aged 8–10 years with different body mass index (BMI) values after supplementation with AmProt. Methods: Participating children were allocated into three groups according to their BMI: normal weight (NW), overweight (OW), and with obesity (OB). Children received AmProt for 90 days. Levels of fasting blood glucose, cholesterol, triglycerides, and insulin were analyzed before and after diet supplementation. HOMA-IR and adinopectin/leptin ratio were evaluated. Feces were collected and metagenome analysis was carried out. Results: No changes in glucose levels were observed across groups and treatments; however, cholesterol and triglycerides levels tended to decrease. The HOMA-IR value increased in relation to BMI and no changes were observed after treatment. Firmicutes were highly abundant in all groups. The lower abundance of Ruminococcus was observed in the OW and OB groups. In the OW group, Blautia, Butyricicoccus, and Roseburia were also observed in increased abundance. In all groups, AmProt consumption tended to increase the abundance of Coproccus, Prevotella, and Collinsella. Conclusions: Supplementation of the children’s diet with AmProt showed an improvement in serum cholesterol and triglyceride levels, which could be related to changes in the microbiota related to lipid metabolism. Full article

Background: Shigellosis is an illness that affects young children all over the world and Shigella flexneri is one of the most common pathogens. The aim of this study was to investigate a potential beneficial effect of Weizmannia coagulans Long45 supplementation on feline-derived Shigella flexneri-induced colitis in mice, as well as potential mechanisms. Results: The results revealed that mice receiving fecal microbiota from diarrheic cats experienced significant weight loss, decreased survival rate, increased mRNA levels of inflammatory cytokines (TNF-α, IL-4, IFN-γ, IL-1β, IL-6, and IL-18), and increased cell apoptosis compared to the single DSS treatment. In contrast, mice that received fecal microbiota from healthy cats exhibited an increased body weight, increased mRNA level of ZO-1, claudin-3, and Muc2 and decreased apoptosis, indicating a protective effect. The 16S rDNA analysis revealed that the abundance of Shigella in the feces of diarrheic cats was significantly higher than that in healthy cats, while the abundance of Bacillus was lower. Using bacteria culture technology, 19 strains of Shigella flexneri were isolated from 27 fecal samples of diarrheic cats and a strain of Weizmannia coagulans Long45 was isolated from the feces of healthy cats. Further study showed that Weizmannia coagulans Long45 significantly alleviated pathological alterations and colonic barrier dysfunction by modulating the NF-κB and Nrf2 signaling pathways. Conclusions: Our data indicate that feline-derived Shigella flexneri may be a potential pathogen associated with diarrhea and intestinal barrier dysfunction. Weizmannia coagulans Long45, as a potential probiotic, can effectively alleviate Shigella-induced colitis by interfering with the Nrf2 and NF-κB signaling pathways. Full article

Ulcerative colitis (UC) is a type of inflammatory bowel disease without curative therapeutics. Recent studies demonstrate that Akkermansia muciniphila exerts mitigating effects on UC, but the underlying mechanisms remain unclear. In this study, we isolated a strain of A. muciniphila, designated NND9, from the feces of DSS-induced ulcerative colitis model mice and investigated its effects on UC of the mouse model. NND9 significantly alleviated UC severity in the mice by restoring gut barrier integrity through improving colonic mucus layer thickness, mitigating goblet cell depletion, and halting epithelial cell death. Mechanistically, NND9 suppressed the expression of the Tnfrsf10b gene encoding death receptor 5 (DR5) on the surface of colonic epithelial cells. Additionally, NND9 inhibited the phosphorylation of kinase 3 (RIPK3) and the pseudokinase mixed-lineage kinase domain-like protein (MLKL) associated with the necrotic apoptosis pathway, thereby reducing gut epithelial cell death. NND9 also markedly ameliorated the gut microbiome of the colitis mice. Untargeted metabolomics analysis demonstrated that NND9 modulated both tryptophan and bile acid metabolism. In conclusion, NND9 exhibits curative effects on UC by resolving inflammatory reactions of the gut mucosa through the DR5-RIPK3/p-RIPK3-MLKL/p-MLKL pathway and redressing gut dysbiosis. This study provides valuable information for the development of innovative therapeutic strategies for the treatment of UC. Full article

Doxorubicin is an effective chemotherapeutic agent, but it causes gastrointestinal toxicity that impairs treatment efficacy and quality of life. This study investigated the effects of liraglutide, a GLP-1 analog, on acute doxorubicin-induced gut toxicity in rats. Sixty male Wistar rats were assigned to four groups: Control (C), Doxorubicin (D), Liraglutide (L), and Doxorubicin + Liraglutide (DL). Groups L and DL received liraglutide (0.6 mg/kg, s.c.) for two weeks. D and DL were given a single dose of doxorubicin (20 mg/kg, i.p). After 48 h, the distal colon, feces, and blood were collected. Results: Doxorubicin caused crypt disruption, goblet cell loss, apoptosis, and reduced fecal short-chain fatty acids. Levels of TNF-α, NF-κB, Bcl-2, TLR4, and antioxidant enzymes were unchanged among groups. Microbiota analysis showed similar α-diversity but altered β-diversity. Doxorubicin reduced Bacteroidetes and increased Proteobacteria, with higher Arcanobacterium and Clavibacter genera abundance. Liraglutide alone decreased Bacteroidetes and increased Corynebacterium and Actinobaculum genera. Combined treatment showed no significant effects. We conclude that acute doxorubicin administration induces intestinal structural damage, reduces short-chain fatty acids, and changes microbiota composition. Although liraglutide alters microbial profiles, it does not attenuate doxorubicin-induced gut toxicity. Full article

Linezolid represents a critical last-resort treatment for severe multidrug-resistant (MDR) Gram-positive bacterial infections. Rising linezolid resistance in Enterococcus isolates threatens its efficacy; this study characterized the molecular features and transfer potential of plasmid-encoded linezolid resistance genes optrA and poxtA in a linezolid-resistant Enterococcus asini isolate from chickens. An E. asini strain was isolated during a surveillance program focusing on drug-resistant Gram-positive bacteria in poultry. PCR screened linezolid resistance genes, conjugation and plasmid stability assays evaluated gene transferability and stability, and whole-genome sequencing (WGS) was performed using both the Illumina and Nanopore platforms. We present the first detection of optrA and poxtA genes in E. asini recovered from chicken feces in China. Sequence analysis of the complete genome showed that poxtA and optrA were situated on two distinct plasmids. The poxtA positive plasmid, pHNGXN23C145Ea-1, also carried multiple resistance genes, including tet(S), fexB, erm(B), ant(6)-Ia, aph(3′)-III. Furthermore, the poxtA gene was flanked by IS1216E mobile elements. The optrA bearing plasmid, pHNGXN23C145Ea-2, harbours a common genetic array of ‘IS1216E fexA-optrA-erm(A)-IS1216E’. Conjugation experiments indicated that neither the poxtA- nor the optrA-bearing plasmid was transferred to recipient strains, which was consistent with sequence analysis showing that both plasmids lacked intact conjugative transfer regions. Stability assays confirmed that poxtA and optrA remained highly stable in the absence of selective pressure. Notably, this discovery was made in a livestock sample, despite the non-use of linezolid in food animals, suggesting that such niches may act as silent reservoirs for resistance genes, which could persist and potentially transfer to clinically relevant MDR pathogens. Full article

Kennel-housed dogs may experience chronic stress affecting cognition. This study compared the effects of Lycium barbarum polysaccharides (LBP) and glycopeptide (LbGP) on cognitive function in laboratory-kenneled poodles. Eighteen dogs were assigned to CON, LBP, or LbGP groups for 42 days. Cognitive tests were performed and serum, saliva, and feces were collected for subsequent analysis. Both supplements improved cognitive performance (e.g., increased the correct rate in the cylinder test by approximately 40.0%, both p < 0.001). LBP enriched beneficial bacteria (Faecalibacterium and Bacteroides, p < 0.05), reduced pathogens (Romboutsia and Terrisporobacter, p < 0.05), and predominantly influenced the indole pathway of tryptophan metabolism. LbGP specifically decreased Escherichia-Shigella and Corynebacterium, increased fecal SCFAs, and mainly targeted the 5-HT pathway. Both treatments regulated immune function (i.e., elevated IL-6 and IL-10) and antioxidant capacity; LBP significantly increased serum superoxide dismutase (SOD) levels by 6.8% (p < 0.01) and BDNF levels by 13.5% (p < 0.05), while LbGP elevated (p < 0.01) glutathione peroxidase (GSH-Px) levels by 20.9% and reduced salivary cortisol levels by 14.2% (p < 0.01). Overall, LBP and LbGP support canine cognition through distinct microbiota-related mechanisms, likely via the microbiota–gut–brain axis, suggesting their potential as functional feed additives for cognitive health. Full article

Klebsiella pneumoniae has been associated with reproductive infections in equines. The detection of β-lactam resistance determinants, especially extended-spectrum β-lactamase (ESBL) genes, within genomic regions linked to horizontal gene transfer (HGT), is of a particular concern. In this study, we characterize the whole-genome sequences (WGS) of three K. pneumoniae equine isolates harboring multiple antimicrobial resistance genes. Two isolates were recovered from uterine washes of mares: one with endometritis (YAH-KPEM1) and one clinically normal (YAH-KPSE1), and a third from the feces of a diarrheic foal (YAH-KPF132). WGS was performed using the Illumina MiSeq platform, and the reads were subsequently processed through hybrid assembly in Unicycler v0.5.1. Genome annotation was completed using PROKKA v1.14.5. Strain YAH-KPEM1 was classified as ST127, whereas YAH-KPSE1 and YAH-KPF132 belonged to ST1630 and ST224, respectively. Notably, K. pneumoniae ST1630 and ST224 have not been reported before in equines. All three genomes encoded multiple antimicrobial resistance (AMR) determinants, including two encoding ESBL genes (CTX-M-15), as well as virulence factors and regions associated with HGT. Additionally, two (YAH-KPEM1 and YAH-KPSE1) isolates were found to be multidrug resistant (MDR), harboring an IncFIB(K) plasmid replicon, and another isolate, YAH-KPF132, carried an IncFII replicon. The detection of AMR and virulence genes in equine Klebsiella isolates has important clinical implications for guiding antimicrobial selection and improving treatment success. Full article

This study aimed to evaluate the potential of Lonicera japonica Flos (LJF) as an alternative agent against Eimeria tenella (E. tenella) in chickens and to conduct phytochemical analysis to obtain compositional insights. Seventy-two one-day-old chickens were allocated to six groups and fed diets supplemented with LJF powder (LJFp) at three concentrations (LJFp-L, LJFp-M, and LJFp-H) or maduramicin (MDM) or two non-supplemented control diets, namely, an infected unmedicated control (IUC) and an uninfected unmedicated control (UUC). Dietary treatments were initiated at chick arrival (Day 0) and continued for 28 days. At 21 days of age, all groups except the UUC group were orally challenged with a field isolate of E. tenella (PT-Te003; 2.0 × 10⁴ oocysts/bird). Anticoccidial efficacy was assessed using the lesion score (LS), oocysts per gram of feces (OPG), relative body weight gain (rBWG), and anticoccidial index (ACI). The results demonstrated that all LJFp treatment groups had significantly reduced cecal OPG and LS (all LJFp treatments: p < 0.05 vs. IUC), indicating the effective suppression of E. tenella replication and intestinal damage. Regarding growth performance, the rBWG values of the LJFp-L and LJFp-M groups were the highest and comparable to those of the UUC group, showing no significant differences. In contrast, the LJFp-H and MDM groups exhibited significantly lower values (p < 0.05). Based on ACI evaluation, all LJFp-treated groups exhibited moderate to partial efficacy (LJFp-L > LJFp-M > LJFp-H), while MDM showed limited effectiveness. A gas chromatography–mass spectrometry (GC-MS) analysis of the LJFp ethanol extract revealed 15 essential oils, 10 organic acids, and three other compound classes, several of which have been associated with anticoccidial activity. Overall, the in vivo results suggest that LJF may exert potential anticoccidial effects against a field isolate of E. tenella. Phytochemical analysis provided preliminary compositional insights, and further studies are warranted to optimize extraction methods and evaluate efficacy at lower concentrations under additional in vitro and in vivo conditions. However, the current evidence remains insufficient to determine whether the field isolate exhibits reduced sensitivity to commercially available anticoccidial drugs, and additional studies are needed to clarify this issue. Full article

Background/Objectives: Enteropathogenic Escherichia coli O157:H7 infection disrupts intestinal homeostasis, causing dysbiosis, barrier dysfunction, and inflammation. This study aimed to evaluate the protective efficacy and mechanisms of a novel probiotic, Bacteroides thetaiotaomicron type strain ATCC 29148, isolated from goat feces, against E. coli O157:H7-induced colitis. Methods: This study assessed the protective potential of the probiotic strain Bacteroides thetaiotaomicronBT6 and BT7 in vitro for GI tolerance, adhesion, and no adverse effects were observed. For the in vivo experiment, male C57BL/6J mice were divided into groups treated with Bacteroides thetaiotaomicron (BT6), PBS, E. coli O157:H7, or a combination. We employed integrated analyses including 16S rRNA gene sequencing, antioxidant status, cytokine profiling, and short-chain fatty acid (SCFA) measurement. Results: In vitro, Bacteroides thetaiotaomicron (BT6 and BT7) showed high gastrointestinal tolerance (71.89–93.22% survival). In vivo, it significantly mitigated infection-associated weight loss and disease activity (p < 0.05). Probiotic treatment enhanced barrier integrity, reduced colonic inflammation, and modulated systemic immune responses, notably increasing anti-inflammatory IL-10 while decreasing pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 (p < 0.05). It also alleviated oxidative stress by reducing malondialdehyde (MDA) and elevating antioxidant enzymes (SOD, CAT, GSH) and ATP. Fecal SCFA profiling revealed increased propionic and butyric acid. 16S sequencing indicated that B. thetaiotaomicron (BT6) administration increased beneficial families (Lactobacillaceae, Muribaculaceae) and suppressed pathobionts. Conclusions: B. thetaiotaomicron (BT6) probiotic with potential for mitigating enteropathogenic infection, an effect mainly determined by its capacity to reestablish the intestinal epithelial barrier and enhance global host health, and modulating the inflammatory response. Full article

Background and Objectives: Sepsis-associated acute kidney injury (SA-AKI) is driven by exaggerated inflammation and oxidative stress, with the HMGB1–RAGE axis playing a pivotal role in amplifying tissue damage. This study aimed to investigate the renoprotective effects of papaverine in a feces-induced peritonitis (FIP) model of sepsis and to explore its impact on HMGB1–RAGE-mediated inflammatory and oxidative pathways. Materials and Methods: Sepsis was induced in male Wistar rats by intraperitoneal injection of fecal slurry (1 g/kg). Animals were treated with papaverine (20 or 40 mg/kg, i.p.) one hour after FIP induction and evaluated at 24 h. Renal function (BUN, creatinine, lactate), inflammatory markers (HMGB1, TNF-α, CRP), oxidative stress (MDA), circulating sRAGE levels, renal NF-κB levels, and histopathological injury scores were assessed. Results: The FIP model resulted in an early mortality rate of 20% and produced marked renal histopathological alterations. Biochemically, FIP increased plasma HMGB1, TNF-α, CRP, MDA, BUN, creatinine, and lactate levels while decreasing sRAGE. Papaverine treatment dose-dependently reduced inflammatory and oxidative markers, restored sRAGE levels, improved renal function parameters, and attenuated histopathological injury. In addition, renal NF-κB levels were significantly elevated in the FIP group compared to controls and were dose-dependently reduced following papaverine treatment. Conclusions: FIP-induced sepsis activates an HMGB1-driven inflammatory–oxidative cascade contributing to SA-AKI. Papaverine confers dose-dependent renoprotection by suppressing HMGB1–RAGE signaling, attenuating NF-κB activation, reducing oxidative stress, and preserving renal structure and function. Targeting the HMGB1–sRAGE axis may represent a promising therapeutic strategy in sepsis-associated renal injury. Full article

Nitrogen (N) utilization by ruminants affects production efficiency, feeding costs, and environmental N losses in confined production systems. Palm kernel cake (PKC), an abundant agro-industrial by-product in tropical regions, has been increasingly used in ruminant diets, although its effects on nitrogen dynamics remain inconsistent. In this study, we systematically reviewed and meta-analyzed the effects of dietary PKC inclusion on N intake, excretion, absorption, and retention in confined cattle, goats, and sheep. Eleven studies published between 1995 and 2025, comprising 44 treatment means and 322 experimental units, were included in the meta-analysis. A random-effects model was applied, and the ruminant species was used as a moderator, defining a significant level at 0.05. Overall, the pooled effects indicated that species significantly influenced N intake (p < 0.01) and N absorption (p < 0.01). Species also showed a tendency to influence N in feces (p = 0.062) and manure N (p = 0.073), whereas N in urine (p = 0.194) and N retention (p = 0.170) were not affected. In subgroup analysis, PKC inclusion reduced N intake in goats (Standardized Mean Difference (SMD)) = −0.792; 95% CI (Confidence Interval) = −1.428 to −0.155; I² (Heterogeneity) = 76.7%) and cattle (SMD = −1.576; 95% CI = −2.250 to −0.902; I² = 65.7%), N in urine in cattle (SMD = −0.478; 95% CI = −0.806 to −0.150; I² = 0%), N absorption (SMD = −0.873; 95% CI = −1.517 to −0.229; I² = 77.1%), and N retention (SMD = −0.875; 95% CI = −1.338 to −0.412; I² = 64.1%) in goats. Conversely, PKC had a positive effect on N absorption in sheep (SMD = 1.137; 95% CI = 0.016 to 2.258; I² = 72.4%). Overall, this study highlights the species-dependent responses of N dynamics to PKC inclusion, emphasizing the importance of species-specific dietary strategies when using agro-industrial by-products to improve nitrogen utilization efficiency and potentially mitigate N losses in confined ruminant systems. Full article