Porcine cysticercosis caused by Taenia solium remains a neglected zoonotic disease in high-altitude Andean regions, where backyard pig-rearing, limited sanitation, and close human–animal–environment interactions promote transmission. Pigs serve as effective sentinels of environmental contamination and offer a practical surveillance proxy for the taeniasis/cysticercosis complex. This study aimed to determine the seroprevalence of porcine cysticercosis, identify associated risk factors, and characterize spatial clustering of infection in backyard pigs in the José María Arguedas district, Apurímac, Perú, using a One Health approach.  A quantitative, observational, analytical, cross-sectional study was carried out from April to December 2024. Blood samples were collected from 245 backyard pigs across 49 households and tested for anti-T. solium antibodies using the enzyme-linked immunoelectrotransfer blot (EITB) assay. Data on pig and owner characteristics, husbandry practices, sanitation, and knowledge of cysticercosis were gathered through structured questionnaires. Geospatial coordinates of households were recorded and analyzed with geographic information systems (GIS) employing inverse distance weighting interpolation to identify infection hotspots. Associations were assessed using both univariate and multivariate logistic regression analyses.  The overall seroprevalence of porcine cysticercosis was 14.7% (36/245; 95% confidence interval [CI]: 10.1–19.3). Significant spatial clustering was seen, with the Checche locality showing the highest seroprevalence at 60% (p < 0.01), indicating a hyperendemic micro-focus. Univariate analysis revealed associations between porcine cysticercosis and the pig owner's sex (p < 0.05) as well as awareness that humans can be infected (p < 0.05). Multivariate logistic regression identified only the pig owner's sex as an independent risk factor, with pigs raised by women having higher odds of infection (Odds ratio = 3.2; 95% CI: 1.3–8.2; p = 0.015). No significant links were found with pig-level characteristics.  This study offers the first combined serological and geospatial assessment of porcine cysticercosis in the José María Arguedas district. The results reveal moderate endemicity, localized transmission hotspots, and an important gender aspect in disease risk. Combining EITB diagnostics with GIS-based mapping within a One Health framework provides a scalable surveillance model to support targeted interventions, health education, especially among women farmers, and sanitation improvements in high-risk Andean areas. 

Saline water use in livestock production is increasingly common in coastal and delta regions due to freshwater scarcity, but prolonged intake may compromise productivity, physiological balance, and immune function in small ruminants. Excessive intake of sodium (Na) and chloride (Cl) can induce osmotic and oxidative stress, which may disrupt hematological homeostasis and growth performance. Antioxidant nutrients such as vitamin E (VitE) and selenium (Se) can enhance cellular protection and immune resilience under environmental stress. This study aimed to evaluate the effects of dietary VitE and Se supplementation on productivity, hematological responses, and selected biochemical parameters in growing goats consuming diluted seawater (DSW) under tropical conditions.  Ten crossbred Boer male goats (6 months old; body weight [BW] 16.66 ± 0.52 kg) were allocated to a completely randomized design with two treatments and five replicates per group over a 5-week experimental period. Goats in the control group received no supplementation, whereas goats in the treatment group received a daily premix providing 60 mg VitE and 0.9 mg Se per head. All goats consumed fresh water during period 1 (P1), followed by 1% DSW during period 2 (P2) and 2% DSW during periods 3 and 4 (P3–P4), before returning to fresh water in period 5 (P5). Dry matter intake (DMI) and water intake (WI) were recorded daily, whereas BW, plasma electrolytes, liver and kidney function indicators, and hematological parameters were measured weekly. Data were analyzed using linear mixed-effects models including treatment, period, and their interaction.  DMI was not significantly affected by DSW concentration or VitE–Se supplementation. WI increased at 1% DSW but declined at 2% DSW, with the supplemented group maintaining higher WI than the control group. BW change decreased in the control group during exposure to 2% DSW, whereas BW remained stable in the supplemented group. DSW increased plasma Na and Cl concentrations and elevated liver enzyme activities, while supplementation attenuated Cl accumulation and moderated Na elevation. Hematological analysis indicated greater leukocyte responses in the control group as DSW salinity increased, whereas the supplemented group maintained more stable hematological profiles. Renal and hepatic indicators remained within physiological reference ranges in both groups.  Dietary VitE and Se supplementation mitigated DSW-associated hematological disturbances and supported WI and BW maintenance in growing goats, suggesting a practical nutritional strategy for saline water–affected production systems.

Native Blackbone chickens (Gallus gallus domesticus) are increasingly consumed in Southeast Asia because of their perceived health benefits. However, information on the epidemiology, antimicrobial resistance (AMR), and genomic characteristics of non-typhoidal Salmonella associated with this traditional poultry system remains limited. This study aimed to determine the prevalence, serovar distribution, antimicrobial susceptibility patterns, and genomic features of Salmonella isolated from feces and meat of Thai native Blackbone chickens.  A prospective longitudinal study was conducted between August 2020 and July 2021 on a small-scale Blackbone chicken farm in Phitsanulok province, Thailand. A total of 2,258 samples, comprising 1,755 fecal and 503 meat samples, were collected from chickens aged 1–50 weeks. Salmonella isolation was performed using standard culture and biochemical methods, with confirmation by invA using polymerase chain reaction. Serovars were determined by slide agglutination. Antimicrobial susceptibility was assessed against 14 antimicrobial agents using disk diffusion and broth microdilution methods. Six representative isolates were subjected to whole-genome sequencing (WGS) for multilocus sequence typing, detection of AMR and virulence genes, plasmid analysis, and phylogenetic comparison.  The overall prevalence of Salmonella was 6.3% (142/2,258), with detection rates of 4.7% in fecal samples and 11.9% in meat samples. Salmonella Bovismorbificans was the predominant serovar (64.1%), followed by S. Corvallis (20.4%) and S. Weltevreden (9.2%). More than half of the isolates (52.8%) exhibited resistance to at least one antimicrobial, most commonly streptomycin (46.5%), while resistance to critically important antimicrobials was not observed. Multidrug resistance (MDR) was rare (1.4%). WGS revealed that S. Bovismorbificans isolates belonged to sequence type ST1499, whereas S. Weltevreden was identified as ST365. Despite limited AMR, all sequenced isolates carried numerous virulence-associated genes linked to adhesion, invasion, and intracellular survival.  Thai native Blackbone chickens harbor non-typhoidal Salmonella, including emerging and potentially invasive lineages, with low levels of MDR but substantial virulence potential. These findings highlight the zoonotic risk posed by traditional poultry systems and underscore the importance of continued surveillance and genomic monitoring to support food safety and One Health strategies. 

Poorly managed dairy farm wastewater is a significant reservoir of antibiotic-resistant bacteria, particularly Escherichia coli, contributing to the environmental spread of antimicrobial resistance (AMR) and posing risks to animal and public health. Conventional wastewater treatment systems are often insufficient to inactivate these resistant organisms. Silver nanoparticles (AgNPs), especially those synthesized by pulsed laser ablation (PLA) in liquid, offer a high-purity, chemical-free nanomaterial with promising antimicrobial properties. This study aimed to evaluate the in vitro antimicrobial efficacy of laser-synthesized AgNPs against antibiotic-resistant E. coli isolated from dairy cattle wastewater within a One Health framework.  Wastewater samples were collected aseptically from 50 smallholder dairy farms in East Java, Indonesia. E. coli isolates were identified using standard cultural, morphological, Gram staining, and biochemical (Indole, methyl red, Voges–Proskauer, citrate) methods. Antibiotic resistance was screened using the Kirby–Bauer disk diffusion method against streptomycin, erythromycin, penicillin, and tetracycline. AgNPs were synthesized via PLA in polyvinylpyrrolidone medium and characterized using transmission electron microscopy, ultraviolet–visible spectroscopy, and Fourier transform infrared spectroscopy. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of AgNPs were determined by broth microdilution and agar subculture methods, respectively, across concentrations ranging from 0.195 to 100 ppm. Statistical analysis was performed using one-way analysis of variance followed by Tukey’s post hoc test at a significance level of p < 0.05.  PLA successfully produced monodisperse AgNPs with a mean diameter of 11.62 ± 1.8 nm and a characteristic surface plasmon resonance peak at 418 nm, confirming high-purity and stability. Twenty antibiotic-resistant E. coli isolates were evaluated. MIC values ranged from 37.5 to 100 ppm, with erythromycin-resistant isolates showing the lowest MICs (45.0 ± 10.5 ppm) and streptomycin-resistant isolates the highest (75.0 ± 33.3 ppm). Most isolates (75%) exhibited MBC values >100 ppm, indicating predominantly bacteriostatic activity at the tested concentrations. No statistically significant differences in MIC values were observed among resistance groups (p > 0.05). A concentration of 62.5 ppm was identified as the most effective inhibitory dose across resistance profiles.  Laser-synthesized AgNPs demonstrated consistent in vitro inhibitory activity against antibiotic-resistant E. coli from dairy wastewater, with an optimal MIC of approximately 62.5 ppm. These findings highlight the potential application of AgNPs as a supplementary control strategy in dairy waste management and AMR mitigation, supporting an integrated One Health approach. 

Kalmyk cattle represent a resilient indigenous beef breed of Russia, valued for their exceptional adaptation to harsh continental climates and growing importance in sustainable beef production. Despite their economic and ecological relevance, genetic determinants underlying meat productivity and quality in this breed remain fragmented across largely regional studies. This systematic review aimed to synthesize available evidence on polymorphisms in four major candidate genes, growth hormone (GH), Thyroglobulin (TG), Leptin (LEP), and Calpain 1 (CAPN1), and to evaluate their distribution, associations with productive traits, and relevance for marker-assisted selection in Kalmyk cattle.  The review was conducted in accordance with PRISMA 2020 guidelines. A comprehensive literature search covering January 2004 to December 2024 was performed using international (PubMed, Scopus, Google Scholar) and Russian (eLibrary.ru) databases. Eligible studies included peer-reviewed articles, dissertations, and conference proceedings reporting primary genotyping data for GH, TG, LEP, and CAPN1 polymorphisms in purebred or crossbred Kalmyk cattle. Data extracted included sample size, geographical origin, genotyping methods, allele and genotype frequencies, and reported genotype–phenotype associations. Due to methodological heterogeneity, a qualitative narrative synthesis was applied.  The synthesis revealed pronounced inter-herd and regional heterogeneity in the frequency of favorable alleles. The GH c.2141C>G polymorphism showed extreme variability, with the desirable VV genotype ranging from 0% to 78.3% across herds. Substantial contrasts were also observed for LEP polymorphisms, where favorable genotypes varied from near absence to dominance within specific populations. For meat quality markers, the TG c.-422C>T and CAPN1 c.4568G>C polymorphisms displayed generally low but highly uneven frequencies. Evidence from selected breeding programs demonstrated that targeted selection can substantially increase the prevalence of desirable alleles and improve growth and carcass traits.  Kalmyk cattle exhibit marked genetic heterogeneity for key meat productivity and quality markers, reflecting founder effects, localized selection, and breeding history. While GH and LEP polymorphisms show strong potential for marker-assisted selection, the low baseline frequency of favorable TG and CAPN1 alleles highlights the need for structured, large-scale genomic strategies. This review provides the first consolidated genetic landscape of meat-related polymorphisms in Kalmyk cattle and establishes a foundation for sustainable, climate-resilient breeding programs. 

The gastrointestinal health of Asian elephants (Elephas maximus) is critically dependent on hindgut microbial fermentation, yet host-specific probiotic strains derived from elephants remain poorly characterized. Although lactic acid bacteria (LAB) are widely recognized for their probiotic benefits, systematic evaluation of elephant-origin LAB, including molecular identification and safety assessment, is limited. This study aimed to isolate LAB from the feces of healthy captive Asian elephants and comprehensively evaluate their in vitro probiotic potential, safety profile, and molecular identity to support the development of host-adapted probiotic candidates.  Fresh fecal samples were collected from 25 clinically healthy captive Asian elephants housed at four elephant camps in Krabi Province, Thailand. LAB were isolated using de Man, Rogosa, and Sharpe agar and subjected to preliminary phenotypic and biochemical characterization. Antimicrobial activity was evaluated against five pathogenic indicator bacteria using the disk diffusion method. Probiotic functional properties were assessed through acid tolerance (pH 3.0), bile salt tolerance (1%), cell surface hydrophobicity, and autoaggregation assays. Safety evaluation included hemolytic activity and antibiotic susceptibility testing. Molecular identification of selected isolates was performed using 16S rRNA gene sequencing followed by phylogenetic analysis.  A total of 195 LAB isolates were recovered, of which 52 exhibited antimicrobial activity against all tested pathogens. Eleven isolates demonstrated superior probiotic attributes, with acid and bile salt survival rates ranging from 74.67%–91.67% and 75.17%–98.15%, respectively. These isolates showed strong antimicrobial activity (inhibition zones 12–15 mm), high cell surface hydrophobicity (74.03%–92.24%), and substantial autoaggregation capacity (70.60%–85.74%). All selected isolates were non-hemolytic and susceptible to clinically relevant antibiotics. Molecular analysis identified seven isolates as Enterococcus faecalis and four as Lactiplantibacillus plantarum. Among them, isolates I9, I56, and I145 (L. plantarum) exhibited the most consistent and robust probiotic characteristics.  This study provides the first molecularly validated and comprehensive in vitro evaluation of probiotic LAB isolated from captive Asian elephants. The identified L. plantarum strains, particularly isolates I9, I56, and I145, demonstrated strong functional and safety profiles, supporting their potential as host-specific probiotics for improving gastrointestinal health and disease management in captive and wild Asian elephants. 

Malignant catarrhal fever (MCF) is a highly fatal disease caused by ovine herpesvirus-2 (OvHV-2), with sheep acting as asymptomatic reservoir hosts. In Indonesia, MCF remains underdiagnosed due to its predominantly subclinical presentation, particularly in regions where mixed ruminant farming is practiced. Riau Province has a large population of Bali cattle, and sheep production is increasing, raising concerns regarding OvHV-2 spillover. This study aimed to determine the molecular prevalence of OvHV-2 infection in cattle and identify individual- and herd-level risk factors associated with the occurrence of MCF in Riau Province, Indonesia.  A cross-sectional study was conducted between August and December 2024 involving 274 beef cattle from 76 farms across five districts of Riau Province. Buffy coat DNA extracted from ethylenediaminetetraacetic acid-anticoagulated blood samples was examined for OvHV-2 using nested polymerase chain reaction (PCR). Epidemiological data were collected through structured questionnaires administered to farmers. The associations between potential risk factors and OvHV-2 positivity were evaluated using chi-square tests and multivariate logistic regression analysis.  OvHV-2 DNA was detected in 33 of 274 cattle, yielding an individual-level prevalence of 12.0% (95% CI: 8.0–16.0%). At the farm-level, 21 of 76 farms (27.6%; 95% confidence interval [CI]: 17.0%–38.0%) had at least one PCR-positive animal. Most infected cattle (n = 32/33) were clinically asymptomatic, emphasizing the silent nature of infection. The multivariable analysis identified cattle aged 2–5 years as having a significantly lower risk of infection (adjusted odds ratio [aOR] = 0.239; p = 0.004). In contrast, farms that practiced mixed rearing of cattle and sheep showed a markedly increased risk of MCF (aOR = 8.218; p = 0.015), as did farms located within 100 m of sheep holdings (aOR = 4.083; p = 0.027).  OvHV-2 infection is endemic but largely subclinical in cattle in Riau Province. Close proximity to sheep and co-rearing practices are the primary drivers of MCF risk. These findings underscore the importance of molecular surveillance and the implementation of biosecurity measures, particularly spatial separation between cattle and sheep, to prevent OvHV-2 transmission and reduce economic losses in susceptible cattle populations. 

The global expansion of antimicrobial resistance (AMR), particularly due to extended-spectrum β-lactamase (ESBL)-producing Escherichia coli, represents a critical threat to food safety and public health. Imported meat products are increasingly recognized as potential vehicles for the transboundary dissemination of multidrug-resistant (MDR) bacteria. However, data on the occurrence and molecular characteristics of ESBL-producing E. coli in imported frozen beef in Saudi Arabia remain limited. This study aimed to determine the prevalence, AMR profiles, and ESBL gene distribution among E. coli isolated from retail imported frozen beef.  A total of 78 imported frozen boneless beef samples were collected from retail shops and supermarkets in Dammam and Al Khobar, Eastern Province, Saudi Arabia. E. coli isolation was performed using Enterobacteriaceae enrichment broth followed by CHROMagar™ E. coli. Presumptive isolates were screened on CHROMagar™ ESBL and subjected to antimicrobial susceptibility testing against 21 antimicrobials using the disc diffusion method. Phenotypic confirmation of ESBL production was conducted using E-test ESBL strips. Molecular detection of ESBL-encoding genes (blaTEM, blaSHV, blaCTX-M-1, blaCTX-M-9, and blaCTX-M-15) was performed by polymerase chain reaction.  From 78 beef samples, 390 E. coli isolates were recovered, of which 361 (92.5%) were presumptive ESBL producers on CHROMagar™ ESBL. Phenotypic confirmation showed that 319/361 (88.3%) isolates were ESBL-producing E. coli. Molecular analysis detected β-lactamase genes in 324/361 (89.7%) isolates, with blaCTX-M-1 and blaCTX-M-15 each identified in 87.5% of isolates. High resistance rates were observed to ampicillin (99.4%), cephalothin (97.2%), cephalexin (91.4%), ceftriaxone (81.9%), and cefotaxime (73.9%). MDR was detected in 97.2% of isolates.  Retail imported frozen beef in Saudi Arabia harbors an exceptionally high burden of MDR ESBL-producing E. coli, predominantly driven by CTX-M-1 and CTX-M-15 enzymes. These findings indicate that international beef supply chains may act as significant reservoirs for high-risk ESBL determinants, underscoring the need for strengthened surveillance, regulatory control, and One Health-based interventions to limit foodborne dissemination of AMR. 

Imerslund–Gräsbeck syndrome (IGS) is a rare autosomal recessive disorder characterized by selective intestinal cobalamin malabsorption in humans and several canine breeds. In Border Collies, IGS is caused by a single cytosine deletion in exon 53 of the CUBN gene (c.8392delC), resulting in a frameshift mutation and loss of Cubam receptor function. Although carrier frequencies have been reported in European and East Asian populations, region-specific genetic epidemiology data from Southeast Asia are lacking. This study aimed to determine the genotypic distribution and allele frequency of the CUBN c.8392delC mutation in Thai Border Collies and to validate a cost-effective molecular diagnostic approach combining double-mismatch allele-specific primers with multiplex polymerase chain reaction (PCR).  A total of 107 clinically healthy Border Collies from private owners and breeding kennels in Thailand were enrolled. Genomic DNA extracted from peripheral blood was genotyped using a newly optimized double-mismatch allele-specific multiplex PCR assay designed to discriminate wild-type and mutant CUBN alleles, with an internal control amplicon included in each reaction. Synthetic mutant DNA fragments were used as positive controls for homozygous mutant detection. All PCR-based genotyping results were independently validated by Sanger sequencing. Genotype and allele frequencies were calculated, and 95% confidence interval (CI) values were estimated using the Wilson score method.  Of the 107 dogs examined, 106 were homozygous wild-type (WT; CUBN +/+) (99.1%; 95% CI: 94.8%–99.8%), one dog was a heterozygous carrier (CUBN +/−) (0.9%; 95% CI: 0.16–5.0%), and no homozygous mutant individuals (CUBN −/−) were identified (0%; 95% CI: 0%–4.3%). The estimated wild-type and mutant allele frequencies were 99.6% (95% CI: 98.2%–99.9%) and 0.4% (95% CI: 0.07%–1.8%), respectively. The double-mismatch allele-specific multiplex PCR assay demonstrated 100% concordance with Sanger sequencing, with no false-positive or false-negative results.  This study provides the first population-level screening data for the CUBN c.8392delC mutation associated with IGS in Thai Border Collies and indicates a very low carrier and mutant allele frequency in this genetically diverse population. The validated double-mismatch allele-specific multiplex PCR assay offers a rapid, reliable, and cost-effective tool for routine genetic screening, supporting evidence-based breeding strategies and future large-scale genetic surveillance of hereditary disorders in dogs. 

Novel variant infectious bursal disease virus (nVarIBDV) occurs worldwide, causing significant morbidity and bursal atrophy in chickens, leading to immunosuppression and heavy economic losses. This study aimed to evaluate the pathogenicity, immunogenicity, and virus load of nVarIBDV in organs of specific-pathogen-free (SPF) chickens.  Sixty, 21-day-old SPF chickens were divided into Groups A and B. Birds in Group A were inoculated with 106.75 EID50/ mL of nVarIBDV (UPM1432/2019) while Group B served as a control. Four birds from Group B were sacrificed at 0 days post-inoculation (dpi). Four birds from each group were sacrificed at 1, 3, 5, 7, 10, 14, and 21 dpi. Clinical signs and serum samples were collected. Body, bursa, and spleen weights, and gross lesions were recorded. Bursa samples were collected for histological examination, while the bursa, spleen, caecal tonsils, thymus, and bone marrow were collected for the virus load determination. Data was analyzed using Student’s t-test at 95% confidence level.  Watery diarrhea and ruffled feathers, bursal atrophy, yellowish stain, decreased folds, firm consistency of bursa, and splenomegaly were observed in Group A. Bursal follicles were atrophied with bursal lesion scores of 4 to 5. IBD antibody titer in Group A, ranging from 6921 -13869 ELISA units at 5-21 dpi, was significantly higher (p<0.05) than in Group B at all timepoints. The viral load was highest in the bursa and lowest in the bone marrow and was detected from 1-21 dpi in the bursa, spleen, caecal tonsil, and thymus, and up to 7 dpi in the bone marrow with a copy number ranging from 7.111 - 12.414 log10.  The nVarIBDV was highly pathogenic, immunogenic, and highly infective in the organs of SPF chickens. It could cause immunosuppression in chickens, exposing them to secondary infections with resultant heavy economic losses, and this report of the detection of nVarIBDV in these organs up to 21 dpi is novel. 

Veterinarians experience disproportionately high levels of psychological distress and suicidal ideation compared with other professional groups. Occupational stress (OS), compassion fatigue, workplace environment (WE), and coping strategies have been identified as relevant risk or protective factors; however, their interrelationships remain insufficiently explored, particularly in Brazil. This study aimed to examine the direct and indirect pathways linking occupational psychosocial factors to suicidal ideation among Brazilian veterinarians working in companion-animal practice, with psychological distress conceptualized as a central mediating mechanism.  A nationwide cross-sectional survey was conducted between June and July 2022 involving 1,472 veterinarians exclusively engaged in small animal practice in Brazil. Participants completed validated self-report measures assessing psychological distress (Kessler-6), suicidal ideation, coping strategies, compassion fatigue, OS, and WE. Data were analyzed using partial least squares structural equation modeling (PLS-SEM) with 5,000 bootstrap resamples. Direct, indirect, and mediating effects were estimated, and model fit, reliability, and validity were assessed. Ethical approval was obtained from the Research Ethics Committee of Inspirar College (protocol no. 31645220.4.0000.5594).  Psychological distress emerged as the strongest predictor of suicidal ideation (β = 0.40; p < 0.001), explaining 20.6% of its variance. OS showed a substantial positive association with psychological distress (β = 0.47; p < 0.001), followed by compassion fatigue (β = 0.15; p < 0.001) and WE (β = 0.07; p = 0.028). Coping strategies exerted a significant protective effect on psychological distress (β = −0.26; p < 0.001) and suicidal ideation (β = −0.08; p = 0.013). Indirect effects confirmed that psychological distress mediated the associations between OS, compassion fatigue, WE, and suicidal ideation. Younger age and female gender were indirectly associated with higher suicidal ideation through increased psychological distress.  Psychological distress plays a central mediating role in the relationship between occupational psychosocial factors and suicidal ideation among Brazilian veterinarians. Interventions targeting OS reduction, emotional support, and the promotion of adaptive coping strategies may substantially mitigate suicide risk in veterinary clinical practice. 

The Alope village chicken is an indigenous Indonesian line developed to combine local adaptability with improved growth potential. However, age-specific physiological and metabolic reference data remain limited, constraining evidence-based feeding and health management. This study aimed to integrate blood biochemical profiling and plasma metabolomics to characterize age-related metabolic changes, establish baseline reference values, and identify putative age-discriminant biomarkers in Alope village chickens.  A cross-sectional design was applied using 30 clinically healthy female Alope village chickens (Gallus gallus domesticus), stratified into three age groups: G1 (8 weeks), G2 (12 weeks), and G3 (16 weeks) (n = 10 per group). Blood samples were collected for biochemical analysis of glucose, total protein, urea, and triglycerides using enzymatic colorimetric assays. Plasma metabolomic profiling was conducted using untargeted gas chromatography–mass spectrometry. Multivariate analyses, including principal component analysis and partial least squares–discriminant analysis, were used to explore age-associated metabolic patterns. Discriminant metabolites were identified based on variable importance in projection values and descriptive receiver operating characteristic analysis.  All biochemical parameters increased significantly with age (p < 0.05), remaining within physiological reference ranges, indicating normal metabolic development. Multivariate metabolomic analysis revealed clear age-dependent separation, with distinct metabolic signatures characterizing each growth stage. Exploratory pathway mapping indicated age-associated involvement of amino acid metabolism (glycine, serine, and threonine), lipid metabolism (glycerolipid and glycerophospholipid turnover), purine metabolism, the citrate cycle, and nodes linked to the pentose phosphate pathway. Seven metabolites demonstrated strong discriminatory performance (variable importance in projection ≈ 1.17–1.35; area under the curve ≥ 0.985) and were identified as putative age-related biomarkers, reflecting coordinated shifts in one-carbon/redox balance, membrane remodeling, and nucleotide and energy metabolism. Age was the primary determinant of metabolic organization in Alope village chickens from 8 to 16 weeks. The integration of blood biochemistry and plasma metabolomics revealed coordinated physiological transitions from early growth toward enhanced energy handling and metabolic stabilization. The identified candidate metabolites provide an exploratory framework for defining physiological age and support the potential application of age-informed metabolic indicators for precision nutrition and health monitoring in indigenous village chicken production systems.

Return-to-estrus (RTE) after insemination is a major source of reproductive inefficiency in commercial swine herds, particularly in primiparous sows that are physiologically distinct from multiparous animals. Under tropical conditions, heat stress, metabolic demands of first lactation, and suboptimal reproductive management may further exacerbate post-weaning fertility problems. However, limited large-scale evidence is available focusing exclusively on first-parity sows. This study aimed to identify reproductive, management, and seasonal risk factors associated with RTE following the first post-weaning insemination in primiparous Landrace × Yorkshire sows raised under tropical conditions.  A retrospective observational study was conducted using farm records from a commercial herd in central Vietnam. Data from 5,111 primiparous sows were included after applying predefined inclusion and exclusion criteria. Explanatory variables comprised age at first insemination, age at first farrowing, number of piglets born alive (NBA), litter birth weight (LBW), litter size and weight at weaning, lactation length (LL), weaning-to-service interval (WSI), age at first insemination after weaning (ASAI), and month of first post-weaning insemination (MSAI). Univariable and multivariable logistic regression analyses were performed to evaluate associations with RTE. Multicollinearity was assessed using Spearman’s correlation coefficients and variance inflation factors. Model fit and discrimination were evaluated using the Hosmer–Lemeshow test and the area under the receiver operating characteristic curve.  The overall RTE rate was 8.6%. In the multivariable model, MSAI, WSI, ASAI, and NBA were significant predictors of RTE. Sows inseminated during December–May had significantly lower odds of RTE than those inseminated in August–September. A WSI of 2–6 days was associated with the lowest RTE, whereas delayed (7–20 days) intervals increased risk. The lowest RTE was observed in sows with ASAI of 380–400 days. An NBA greater than 14 piglets was associated with a reduced likelihood of RTE. LBW, litter size and weight at weaning, and LL were not independently associated with RTE.  RTE in primiparous sows under tropical conditions is strongly influenced by seasonal timing and post-weaning reproductive management. Optimizing insemination timing, maintaining an appropriate WSI, and mitigating heat stress may substantially improve fertility and reduce non-productive days in tropical swine production systems. 

Salmonella spp. are major foodborne pathogens within the poultry chain and pose a substantial public health risk. Rapid and accurate serotype identification is essential for effective surveillance, outbreak investigation, and control strategies. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) provides rapid species-level identification; however, its reliability for serotype differentiation remains limited by overlapping spectral profiles and incomplete reference databases. Concurrently, rising antimicrobial resistance (AMR) necessitates exploration of alternative antimicrobial agents. This study aimed to (i) evaluate the accuracy of MALDI-TOF MS for Salmonella serotype identification using specific peak analysis compared with conventional serotyping and (ii) assess the antimicrobial effectiveness of trans-cinnamic acid (TCA) against poultry-associated Salmonella isolates.  A total of 63 Salmonella isolates representing six serotypes were analyzed, including Salmonella Enteritidis, Salmonella Typhimurium, Salmonella Kentucky, Salmonella Newjersey, Salmonella Fresno, and Salmonella Weltevreden, obtained from poultry production environments in Thailand. MALDI-TOF MS performance was evaluated against conventional serotyping using overall percentage agreement (OPA), positive percentage agreement (PPA), negative percentage agreement (NPA), and Cohen’s kappa statistic. Serotype-specific mass spectral peaks were identified through comparative analysis with reference strains. The antimicrobial activity of TCA was evaluated using broth dilution assays to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC).  MALDI-TOF MS showed high concordance with conventional serotyping, with OPA, PPA, and NPA values ranging from 97.3% to 100%. Cohen’s kappa values indicated substantial to perfect agreement, with minor discordance observed for S. Enteritidis (κ = 0.65). Serotype-associated peaks were consistently detected at 6,094 ± 2 mass-to-charge ratio (m/z) for S. Enteritidis, 7,156 ± 2 m/z for S. Typhimurium, and 5,370 ± 2 m/z for S. Kentucky. TCA exhibited uniform antimicrobial activity against all tested serotypes, with MIC and MBC values of 10 mM and 20 mM, respectively, and no significant differences among serotypes (p > 0.05).  MALDI-TOF MS combined with specific peak analysis provides a reliable and rapid approach for Salmonella serotype identification in the poultry chain, although database expansion remains necessary for uncommon serotypes. TCA demonstrated consistent inhibitory and bactericidal activity, supporting its potential role as a complementary, non-antibiotic intervention for Salmonella control and AMR mitigation in poultry production systems. 

The African Houbara bustard (Chlamydotis undulata), a species of high conservation concern, is maintained through intensive captive breeding programs where bacterial diseases may compromise productivity and reintroduction success. Escherichia coli is frequently implicated in reproductive failure, early mortality, and septicemic conditions; however, integrated phenotypic and genotypic characterization of antimicrobial resistance and virulence across life stages in this species remains scarce. This study aimed to determine the prevalence of E. coli across different life stages of captive African Houbara bustards, characterize multidrug-resistant (MDR), extensively drug-resistant (XDR), and extended-spectrum β-lactamase (ESBL) phenotypes, and profile avian pathogenic E. coli (APEC)–associated virulence genes within a One Health framework.  A cross-sectional diagnostic investigation was conducted during the 2025 production season. A total of 110 samples were collected from early embryonic death (n = 50), chicks (n = 25), juveniles (n = 25), and adult females (n = 10). Isolation and identification of E. coli were performed using conventional bacteriology and the VITEK®2 system. Antimicrobial susceptibility testing was carried out using disk diffusion and VITEK®2 AST, with resistance patterns classified according to international MDR and XDR definitions. Phenotypic ESBL detection was undertaken, and a multiplex real-time polymerase chain reaction (PCR) assay targeting 15 APEC-associated virulence genes was applied to representative isolates.  E. coli was isolated from 62.7% (69/110) of samples, with the highest prevalence observed in juveniles (84%) and chicks (80%). All isolates were resistant to ampicillin and fluoroquinolones, while complete susceptibility was observed to fosfomycin, aminoglycosides, and chloramphenicol. MDR was detected in 96.9% of tested isolates, and one XDR isolate resistant to 15 antibiotics was identified. ESBL-producing E. coli were detected in juveniles (14%) and early embryonic death samples (9%). Virulence profiling revealed a high gene burden, with most isolates harboring ≥10 virulence genes, particularly those associated with iron acquisition, protectins, and invasion.  Captive African Houbara bustards harbor highly virulent MDR and ESBL-producing E. coli across life stages, posing significant conservation, veterinary, and public health risks. Strengthened biosecurity, prudent antimicrobial stewardship, and integrated One Health surveillance are essential to improve breeding success and safeguard reintroduction programs. 

Acid–base disturbances, particularly metabolic acidosis, are common in dogs with advanced renal failure and contribute substantially to morbidity and prognosis. Intermittent hemodialysis (IHD) is increasingly used when conventional therapy fails; however, physiological monitoring parameters for incremental intermittent hemodialysis (i-IHD) in dogs remain poorly defined. Venous blood gas (VBG) analysis offers a safer and more practical alternative to arterial sampling, yet its utility during i-IHD has not been systematically evaluated. This study aimed to characterize longitudinal changes in VBG and hemato-biochemical parameters before and after consecutive i-IHD sessions in dogs with renal failure.  In this prospective observational study, 45 client-owned dogs with severe azotemia (serum creatinine >5 mg/dL) due to acute kidney injury (AKI) stage IV–V or chronic kidney disease (CKD) stage IV were enrolled. All dogs underwent three consecutive i-IHD sessions with stepwise increases in treatment intensity. Venous blood samples were collected immediately before and after each session for VBG analysis, including pH, bicarbonate (HCO₃⁻), total carbon dioxide (TCO₂), partial pressures of carbon dioxide (pCO₂) and oxygen (pO₂), base excess, anion gap, and cerebral oxygen saturation, along with hemato-biochemical profiling. Dialysis adequacy was assessed using Kt/V, urea reduction ratio, and creatinine reduction ratio. Pre- and post-dialysis values were compared using paired statistical analyses.  Dogs exhibited mild-to-moderate metabolic acidosis before i-IHD. Across all sessions, i-IHD produced a consistent and significant correction of acid–base imbalance, evidenced by normalization of pH and marked increases in HCO₃⁻ and TCO₂ (p < 0.01). A modest but significant rise in pCO₂ accompanied bicarbonate repletion, while the anion gap remained within the lower borderline range, indicating non-anion gap metabolic acidosis. Significant reductions in blood urea nitrogen and creatinine (approximately 25%–40% per session) confirmed effective solute clearance, with adequacy indices improving progressively across sessions. Electrolyte abnormalities, particularly hyperkalemia, were effectively corrected.  I-IHD effectively restores acid–base and biochemical homeostasis in dogs with advanced renal failure. Serial VBG monitoring provides clinically meaningful, session-wise information and represents a practical tool for guiding i-IHD without the risks of arterial sampling. 

Q fever, caused by Coxiella burnetii, is a globally distributed zoonosis with major public health and livestock production implications. Dromedary camels (Camelus dromedarius) are increasingly recognized as important reservoirs, particularly in arid and semi-arid regions. In Jordan, however, comprehensive national-level data integrating serology, molecular detection, and spatial epidemiology are lacking. This study aimed to estimate the serological and molecular prevalence of Q fever in camels, identify associated risk factors, and describe the spatial distribution of infection across Jordan.  A nationwide cross-sectional study was conducted between July and October 2022 using a two-stage cluster sampling design. A total of 468 camels from 31 villages across all camel-rearing governorates were sampled. Serum samples were tested for anti-C. burnetii antibodies using a commercial indirect multi-species enzyme-linked immunosorbent assay, while whole blood was examined for C. burnetii DNA using conventional polymerase chain reaction (PCR) assays targeting IS1111 and CB-1 genes. Survey-weighted prevalence estimates were calculated, and risk factors were evaluated using univariate and multivariate survey-weighted logistic regression. Spatial distribution was mapped at the governorate level.  The weighted prevalence of Q fever was 88.75% (95% confidence interval: 79.26%–98.23%) using parallel interpretation of serological and molecular tests. Individually, prevalence was 44.02% by enzyme-linked immunosorbent assay and 68.93% by PCR. Significant risk factors included age ≥3 years, contact with other camel herds, Sofor breed, light or absent tick infestation, and improper disposal of abortion materials. Co-herding with other species and a history of abortion were associated with reduced odds of positivity. Marked spatial heterogeneity was observed, with the highest prevalence in Tafilah governorate and the lowest in Zarqa.  This first nationwide, integrated serological, molecular, and spatial study demonstrates an exceptionally high burden of Q fever in Jordanian camels, highlighting their critical role as a reservoir. The findings underscore the urgent need to incorporate camels into national Q fever surveillance and control programs within a coordinated One Health framework involving animal, human, and environmental health sectors. 

Enteric methane emissions from ruminants contribute significantly to greenhouse gas production, prompting research into sustainable feed additives. Macroalgae, rich in bioactive compounds, show promise in modulating rumen fermentation, improving digestibility, and reducing methane output. Indonesia’s diverse native macroalgae remain largely unexplored for these purposes, necessitating comparative evaluation to identify promising species for ruminant nutrition. This study aimed to conduct the first comparative in vitro evaluation of rumen fermentation patterns, digestibility characteristics, gas production kinetics, and enteric methane emissions using 14 native Indonesian macroalgae species, including brown (Sargassum sp., Padina sp., Turbinaria ornata), green (Boergesenia forbesii, Caulerpa racemosa, Ulva lactuca), and red (Palmaria palmata, Gelidium sp., Halymenia durvillei, Gracilaria verrucosa, Eucheuma cottonii, Gracilaria gigas, Eucheuma spinosum, Gracilaria coronopifolia) algae, to identify candidates for sustainable ruminant feed additives.  Macroalgae samples were collected from various Indonesian locations, dried, and analyzed for chemical composition (dry matter, ash, crude protein, crude fat, crude fiber, nitrogen-free extract). In vitro fermentation was performed using a completely randomized design with five replicates per species. Samples (0.5 g) were incubated at 39°C for 72 h in buffered rumen fluid from fistulated Ongole crossbreed cattle. Parameters measured included total gas production, methane emissions (estimated via volatile fatty acid [VFA] profiles), ammonia, total and partial VFAs (acetate, propionate, butyrate, valerate, iso-butyrate, iso-valerate), acetate-to-propionate ratio, in vitro dry matter digestibility (IVDMD), in vitro organic matter digestibility (IVOMD), partitioning factor, microbial protein synthesis, and gas production kinetics. Data were analyzed using a one way analysis of variance with significance at p < 0.05 or p < 0.01, followed by post-hoc tests.  Chemical composition varied widely; red algae like Palmaria palmata had high crude protein (22.39 % dry matter), while brown algae like Padina sp. were ash-rich (74.39 % dry matter). Total gas production was highest in B. forbesii (54.75 mL; p < 0.01) and lowest in T. ornata (10.94 mL). Methane emissions and methane per incubated dry matter were lowest in Sargassum sp. (1.87 mM and 3.75 mM/g dry matter; p < 0.01), with Sargassum sp. and C. racemosa reducing methane by 71.86 %. Ammonia levels were similar across species (p > 0.05). Total VFA and propionate were highest in H. durvillei and B. forbesii (p < 0.01), with reduced acetate-to-propionate ratios. IVDMD and IVOMD were highest in H. durvillei (81.72 % and 69.53 %; p < 0.01). Gas kinetics showed B. forbesii with the highest asymptote (201.97 mL; p < 0.01) but slowest rate (0.01 mL/h). Positive correlations existed between crude protein and VFA/ammonia, while crude fiber inversely correlated with gas production and digestibility.  H. durvillei emerged as optimal for enhancing rumen fermentation and digestibility, while Sargassum sp. excelled in methane mitigation. These species hold promise as natural additives for reducing environmental impacts in ruminant production, warranting in vivo validation for optimal inclusion rates and long-term effects. 

Houseflies (Musca domestica) are recognized mechanical vectors of foodborne pathogens and antimicrobial-resistant bacteria, particularly in environments with intensive food-handling. Their role in disseminating virulent and multidrug-resistant (MDR) Escherichia coli at the food–environment–human interface remains underexplored in Northern Thailand. This study aimed to determine the prevalence, phylogenetic distribution, virulence gene carriage, antimicrobial resistance profiles, and resistance determinants of E. coli isolated from houseflies collected at food markets in Phayao Province.  A cross-sectional surveillance study was conducted between June and November 2023 in Muang District, Phayao Province. A total of 350 houseflies were collected from meat, vegetable, and cooked-food markets using sticky traps. E. coli isolates were identified by culture and polymerase chain reaction (PCR) targeting the uidA gene. Phylogenetic grouping was performed using chuA, yjaA, and TSPE4.C2. Virulence genes associated with diarrheagenic E. coli were detected by PCR. Antimicrobial susceptibility testing was conducted using the disc diffusion method against 12 antibiotics representing eight antimicrobial classes. Multidrug resistance was defined as resistance to three or more antimicrobial classes. Resistance-associated genes were screened by PCR. Associations were analyzed using the chi-square test.  Overall, 106 E. coli isolates (30.3%) were recovered, with the highest prevalence in meat markets (39.5%) (p < 0.0001). Phylogroup A predominated (49.1%), followed by D (19.8%), B1 (18.9%), and B2 (12.3%). Virulence genes were detected in 69.8% of isolates, with bfpA being the most prevalent (26.4%). Universal resistance to penicillin G and erythromycin was observed, while high resistance rates were noted for ampicillin (66.0%) and tetracycline (35.8%). MDR was identified in 46.2% of isolates and was significantly more common in meat markets (p = 0.0225). The highest MDR prevalence occurred in phylogroup D (57.1%). The most frequently detected resistance genes were blaSHV, ereA, and tetA.  Houseflies from food markets in Northern Thailand harbor virulent and MDR E. coli, highlighting their role as environmental sentinels and potential disseminators of antimicrobial resistance. These findings underscore the need for improved market hygiene and incorporation of insect vectors into One Health–based antimicrobial resistance surveillance strategies. 

Animal cruelty is a persistent concern for animal welfare and society worldwide. In Thailand, the Prevention of Cruelty to Animals and Animal Welfare Act B.E. 2557 (2014) provides a legal framework to address animal abuse; however, effective enforcement remains constrained by limitations in investigative procedures and the limited integration of veterinary forensic science. Veterinarians from the Department of Livestock Development (DLD) play a central role in responding to cruelty reports, conducting inspections, and supporting legal processes. This study aimed to explore the attitudes of DLD veterinarians toward companion animal welfare, the effectiveness of existing animal welfare legislation, and the current and potential role of forensic science in animal cruelty investigations in Thailand.  A cross-sectional qualitative study was conducted between August 2024 and December 2024. Eighteen veterinarians with DLD from six geographic regions of Thailand were recruited using stratified purposive sampling. Data were collected via a validated, open-ended online questionnaire that addressed animal welfare challenges, legal enforcement, investigative procedures, and forensic applications. Qualitative content analysis was performed using the NVivo software, with double-blind coding to enhance analytical rigor. The codes were organized into categories and overarching themes following inductive thematic analysis.  Multiple interrelated drivers of companion animal cruelty were identified by veterinarians. The major welfare-related themes included a lack of owner responsibility and preparedness, insufficient knowledge and ethical awareness regarding animal care, socioeconomic constraints, deficiencies in stray animal management systems, weaknesses in law enforcement, and cultural attitudes that undermine animal welfare. While most respondents perceived the existing animal cruelty law as partially effective in reducing abuse, they highlighted critical gaps, including ambiguous legal definitions, inconsistent enforcement, and limited public awareness. The absence of standardized veterinary forensic protocols, limited forensic training, and restricted inspection authority were the primary factors driving investigative challenges. Respondents strongly supported the integration of forensic science, emphasizing its potential to improve evidentiary reliability, strengthen legal credibility, and enhance investigative outcomes, while also identifying the need for structured training programs and interagency collaboration.  Companion animal cruelty in Thailand is driven by multifactorial welfare, legal, and societal challenges. Although current legislation provides a foundation for protection, the absence of standardized forensic practices limits investigative effectiveness. Strengthening veterinary forensic capacity through harmonized protocols, targeted training, and coordinated enforcement could substantially enhance animal welfare protection and support consistent application of animal cruelty legislation within a One Welfare framework.

The global restriction on antibiotic growth promoters (AGPs) in poultry production due to antimicrobial resistance concerns has accelerated the search for effective, sustainable alternatives. Probiotics derived from agricultural by-products offer a promising strategy to enhance gut health and productivity while reducing environmental waste. Coconut-waste, including coconut water and pulp, is rich in fermentable substrates that support the growth of lactic acid bacteria (LAB) and the production of functional metabolites. This study aimed to perform an integrated in vitro characterization of indigenous microorganisms derived from coconut-waste fermentation as potential probiotic and synbiotic candidates for sustainable poultry production.  Indigenous microorganism solutions (IMOS) were produced through anaerobic fermentation of coconut water and coconut pulp for 5, 10, 15, and 20 days using a completely randomized design with four treatments and five replicates. Physicochemical properties (pH, LAB counts), enzymatic activities (cellulase and mannanase), tolerance to simulated gastrointestinal conditions (acidic pH 2.5, bile salts at 0.3% and 0.5%, and thermal exposure at 42°C), cell surface hydrophobicity, antimicrobial activity against Escherichia coli, Salmonella spp., and Staphylococcus aureus, and short-chain fatty acid (SCFA) production were evaluated using standard microbiological and analytical methods.  Fermentation duration significantly influenced all evaluated parameters (p < 0.05). IMOS fermented for 15 days exhibited the lowest pH (3.19 ± 0.02), the highest LAB population (2.05 ± 0.13 × 10¹¹ CFU/mL), optimal cellulase (12.50 ± 0.15 U/mL) and mannanase activities (20.48 ± 0.13 U/mL), and the greatest cell surface hydrophobicity (95.09 ± 0.35%). LAB survival remained high under simulated gastrointestinal stress, reaching 80.23 ± 4.12% at pH 2.5 (6 h), 71.45 ± 0.56% in 0.5% bile salts, and 8.09 ± 0.35 × 10¹¹ CFU/mL at 42°C. Antimicrobial assays demonstrated complete inhibition of E. coli after 24 h at 15 days of fermentation. Acetate (3.34–3.43 g/L) and butyrate (0.66–0.71 g/L) were the dominant SCFAs detected.  Coconut waste–derived IMOS demonstrates strong in vitro probiotic and synbiotic characteristics and represents a low-cost, environmentally sustainable functional feed additive for poultry. Fermentation for 15 days yielded optimal functional properties. Further in vivo validation is warranted to confirm efficacy under practical production conditions. 

The fatty acid-binding protein 4 (FABP4) gene is a key regulator of lipid metabolism and has been linked to carcass and meat quality traits in several cattle breeds. However, genetic variation in FABP4 and its phenotypic relevance in Bali cattle (Bos javanicus), an important indigenous Indonesian breed, remains poorly characterized. This study aimed to identify novel single-nucleotide polymorphisms (SNPs) in the FABP4 gene of Bali cattle and to evaluate their associations with in vivo ultrasound-measured carcass traits and fatty acid composition.  Blood samples were collected from 95 Bali cattle bulls sourced from a single population. Genomic DNA was extracted, and a 721-bp fragment of FABP4 spanning intron 2 to exon 4 was amplified and sequenced using the Sanger method. SNP detection and sequence alignment were performed using standard bioinformatics tools. Carcass and meat quality traits, including longissimus dorsi thickness (LDT), backfat thickness (BFT), marbling score (MS), and intramuscular fat (IMF), were measured in live animals using ultrasound imaging. Fatty acid composition was determined from post-mortem muscle samples using gas chromatography–flame ionization detection. Associations between FABP4 genotypes and phenotypic traits were analyzed using a general linear model, with statistical significance set at p < 0.05.  Four novel FABP4 SNPs were identified: g.4631T>C, g.4724T>C, and g.4769G>A located in intron 3, and g.5002C>T located in exon 4. The g.5002C>T variant was a nonsynonymous substitution resulting in a valine-to-alanine amino acid change. Among the identified polymorphisms, only g.5002C>T showed a significant association with LDT (p = 0.0296). Animals carrying the CT genotype exhibited greater LDT compared with CC and TT genotypes. No significant associations were observed between any FABP4 SNPs and BFT, MS, IMF, or fatty acid composition.  This study provides the first evidence of FABP4 genetic variation in Bali cattle and demonstrates that the g.5002C>T polymorphism is significantly associated with LDT. The CT genotype at this locus may serve as a potential genetic marker for improving muscle development in Bali cattle, subject to validation in larger and independent populations. 

Bacterial contamination during liquid storage of boar semen negatively affects sperm quality and fertility outcomes, necessitating the routine use of antibiotics in semen extenders. However, increasing concerns regarding antimicrobial resistance have encouraged the development of alternative antimicrobial strategies. Synthetic antimicrobial peptides (AMPs) have demonstrated broad-spectrum antibacterial activity and may serve as potential substitutes or adjuncts to conventional antibiotics. This study aimed to evaluate the antimicrobial efficacy of the synthetic peptide PA-13 alone and in combination with reduced gentamicin concentrations against Escherichia coli isolated from boar semen, as well as its effects on semen quality during storage at 18°C.  Two experiments were conducted. In Experiment I, fresh semen samples collected from seven healthy adult boars were diluted in Beltsville Thawing Solution supplemented with gentamicin (200 μg/mL, positive control), without antibiotics (negative control), or PA-13 at concentrations of 62.5, 31.25, and 15.625 μg/mL. Total bacterial counts were measured at 0, 24, 48, and 72 h of storage, while semen quality parameters, including sperm motility, viability, acrosomal integrity, and mitochondrial membrane potential, were evaluated on days 1, 3, and 5. In Experiment II, isolated E. coli was incubated with PA-13 alone or in combination with varying gentamicin concentrations, and bacterial growth was monitored over 24 h using optical density measurements.  PA-13 effectively inhibited bacterial proliferation in extended semen during the first 24 h of storage, with lower concentrations (15.625 and 31.25 μg/mL) showing greater antibacterial activity than the higher concentration (62.5 μg/mL). Semen quality parameters were comparable among groups on day 1; however, prolonged storage demonstrated that the highest PA-13 concentration negatively affected sperm motility and viability. Lower PA-13 concentrations preserved semen quality similar to that of the gentamicin-treated control. In Experiment II, combination treatments exhibited synergistic effects, with PA-13 at 3.906 μg/mL combined with gentamicin at 100 μg/mL inhibiting E. coli growth equivalently to gentamicin at 200 μg/mL alone.  PA-13 effectively controls bacterial contamination in stored boar semen while maintaining semen quality at appropriate concentrations. Its combination with gentamicin enables a substantial reduction in antibiotic dosage without compromising antibacterial efficacy. These findings support the use of AMPs as alternative or complementary antimicrobial agents in semen extenders to reduce antibiotic use in the swine industry. 

Agro-industrial byproducts, including distillers dried grains with solubles, sunflower seed meal, palm kernel cake, sweet orange peel meal, brewers’ dried grain, and various fruit and vegetable processing wastes, represent abundant, low-cost alternatives to conventional feed ingredients such as corn and soybean meal in broiler chicken diets. Their incorporation supports sustainability by reducing feed costs, alleviating food-feed competition, and promoting circular bioeconomy principles through waste valorization. However, the presence of complex non-starch polysaccharides, lignocellulosic structures, phytate, mannans, and other antinutritional factors often limits nutrient digestibility, impairs intestinal health, and compromises broiler growth performance when these byproducts are included at higher levels. Exogenous enzyme supplementation, particularly phytase, protease, xylanase, β-glucanase, β-mannanase, cellulase, and multi-enzyme complexes, has emerged as an effective strategy to overcome these limitations. Enzymes hydrolyze indigestible components, reduce digesta viscosity, improve nutrient availability (dry matter, crude protein, energy, and phosphorus), enhance intestinal morphology (increased villus height to crypt depth ratio), modulate gut microbiota toward beneficial populations such as Lactobacillus and Bifidobacterium, and mitigate inflammatory responses. These improvements enable substantially higher inclusion levels of byproducts, up to 50% substitution of conventional ingredients in some cases, without negative effects on body weight gain, feed conversion ratio, or overall performance. Reported performance gains include 1%–16% increases in weight gain, 2%–11% in feed intake, and 1%–26% reductions in feed conversion ratio, depending on byproduct type, enzyme combination, and inclusion level. Economically, enzyme supplementation often offsets its initial cost through better feed efficiency, resulting in lower production cost per kilogram of broiler meat (reductions of 7%–12% in several studies). Environmentally, the approach decreases reliance on high-carbon-footprint crops, reduces manure emissions, lowers greenhouse gas contributions from feed production, and supports waste minimization. Challenges remain, including variability in byproduct composition, enzyme specificity and stability, seasonal quality fluctuations, and occasional inconsistent results across trials. This review concludes that strategic enzyme supplementation offers a practical, science-based pathway to increase the sustainable and efficient utilization of agro-industrial byproducts in broiler production. Future efforts should focus on tailored multi-enzyme formulations, integration of omics technologies for precise matching of enzymes to specific byproducts, and large-scale commercial validation to facilitate wider industry adoption. 

The growing burden of infectious diseases and antimicrobial resistance (AMR) in aquaculture demands safe, host-adapted alternatives to antibiotics. Probiotics derived from the gastrointestinal tract (GIT) of the target host are considered more ecologically compatible and effective than non-host strains. This study aimed to isolate and characterize indigenous gut bacteria from wild Nile tilapia (Oreochromis niloticus) collected from Lake Naivasha, Kenya, to evaluate their probiotic attributes, assess pathogenicity, and identify promising candidates using conventional phenotypic methods and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).  Thirty-eight apparently healthy O. niloticus were sampled, and bacteria were isolated from the entire GIT using standard bacteriological techniques. Fifty autochthonous isolates were obtained and subjected to stepwise screening, including hemolytic activity, antibiotic susceptibility, enzymatic activity (protease and amylase), tolerance to low pH and bile salts, adhesion to stainless steel surfaces, growth kinetics, and in vivo pathogenicity in O. niloticus. Species-level identification was performed using biochemical tests and MALDI-TOF MS.  Of the 50 isolates, 10 (20%) were nonhemolytic and sensitive to at least eight antibiotics. Functional screening reduced these to four candidates exhibiting enzymatic activity, acid and bile tolerance, and adhesion. Three isolates, identified as Rossellomorea marisflavi, Micrococcus luteus, and Paucilactobacillus vaccinostercus, were nonpathogenic to O. niloticus. In contrast, Aeromonas ichthiosmia, despite exhibiting several probiotic-like traits in vitro, caused 80% cumulative mortality and was excluded. Among the nonpathogenic isolates, P. vaccinostercus demonstrated the strongest overall probiotic profile, including superior acid and bile tolerance, high enzymatic indices, robust adhesion (~4.7 × 10⁴ CFU/mL), and favorable growth kinetics.  The gut microbiota of wild O. niloticus from Lake Naivasha harbors a limited but valuable pool of indigenous bacteria with probiotic potential. This study provides the first evidence that P. vaccinostercus is a promising, nonpathogenic probiotic candidate for tilapia aquaculture. These findings support the development of locally adapted, antibiotic-free probiotic strategies to enhance fish health and sustainable aquaculture in Kenya. Further in vivo feeding trials and genomic safety assessments are warranted. 

Calcium source and bioavailability are critical determinants of nutrient utilization, gut microbial ecology, and future productivity in laying hens, particularly during the pre-laying period. Organic calcium salts may exert additional functional effects through microbiota modulation beyond mineral supply alone. This study evaluated the effects of replacing calcium carbonate with calcium citrate on nutrient digestibility, cecal microbiota composition, short-chain fatty acid (SCFA) production, and predicted microbial metabolic pathways in pre-laying hens.  Sixty Hisex Brown pre-laying hens (13–20 weeks of age) were allocated to two dietary treatments: a control diet containing calcium carbonate and an experimental diet in which calcium carbonate was fully replaced with calcium citrate. Diets were formulated to be isocaloric, isonitrogenous, and equal in total calcium content. Nutrient digestibility coefficients were determined using a physiological balance trial. Cecal SCFA concentrations were quantified by gas chromatography. Cecal microbiota composition was analyzed by 16S rRNA gene sequencing, and functional pathway prediction was performed using Kyoto Encyclopedia of Genes and Genomes-based bioinformatic analysis. Statistical significance was set at p < 0.05.  Replacement of calcium carbonate with calcium citrate significantly increased the digestibility of crude fat (+28.7%, p ≤ 0.001), crude protein (+7.29%, p ≤ 0.001), calcium (+7.56%, p ≤ 0.05), and phosphorus (+2.92%, p ≤ 0.05). Cecal concentrations of propionic, butyric, and valeric acids were significantly higher in the calcium citrate group (p ≤ 0.001). Microbiota analysis revealed a higher relative abundance of Bacillota, particularly Lactobacillaceae and Oscillospiraceae, and a reduced proportion of Bacteroidota, including Alistipes. Alpha diversity indices were higher in the experimental group. Functional prediction indicated enrichment of microbial genes associated with carbohydrate, amino acid (phenylalanine, tyrosine, tryptophan), and fatty acid metabolism, alongside reduced methane metabolism.  Dietary calcium citrate markedly improves nutrient digestibility and beneficially reshapes cecal microbiota composition and function in pre-laying hens. These findings highlight calcium citrate as a promising nutritional strategy to enhance gut health, mineral utilization, and feed efficiency, with potential implications for subsequent egg production and sustainable poultry systems. 

Hepatic lymph nodes (HLNs) drain lymph from the liver, gallbladder, pancreas, stomach, and duodenum. Enlargement of these nodes may indicate pathology in those organs, but objective, breed-independent computed tomography (CT)-based size criteria have not been established. This study aimed to determine quantitative cutoff values for HLN dimensions normalized to aortic diameter (Ao) to differentiate normal from abnormal HLNs in dogs.  This retrospective study analyzed contrast-enhanced abdominal CT scans (slice thickness <1.5 mm) from 84 dogs older than 1 year, examined between March 2016 and November 2022. Dogs were divided into two groups: no targeted organ abnormalities (26 dogs; 46 HLNs) or presence of targeted organ abnormalities in the liver, gallbladder, pancreas, stomach, duodenum, or combinations (58 dogs; 89 HLNs), based on clinical, laboratory, and imaging findings. Using multiplanar reconstruction, the maximum length (cranio-caudal), width (latero-lateral), and height (dorso-ventral) of each HLN were measured and divided by the Ao measured at the same level. Parameters were compared between groups using Mann–Whitney U tests. Receiver operating characteristic curves were used to identify optimal cutoff values, sensitivity, specificity, and area under the curve (AUC).  HLN dimensions and dimension-to-Ao ratios were significantly larger in dogs with targeted organ abnormalities (all p < 0.001). Mean width-to-Ao ratio, length-to-Ao ratio, and height-to-Ao ratio were 0.7 ± 0.3, 1.4 ± 0.7, and 0.6 ± 0.3 in the normal group versus 1.2 ± 1.2, 2.0 ± 0.8, and 0.8 ± 0.4 in the abnormal group, respectively. The width-to-Ao ratio showed the highest diagnostic performance (AUC = 0.76, 95% confidence interval: 0.69–0.85; p < 0.0001), with an optimal cutoff of ≥ 0.75 (sensitivity 70%, specificity 67%). Height-to-Ao ratio cutoff was ≥ 0.615 (AUC = 0.70), and length-to-Ao ratio cutoff was ≥ 1.58 (AUC = 0.70).  Ao-normalized HLN dimension ratios provide an objective, breed- and body-size-independent criterion for CT evaluation of canine hepatic lymph nodes. A width-to-Ao ratio ≥ 0.75 offers the best balance of sensitivity and specificity for identifying HLN abnormalities suggestive of targeted organ pathology. These cutoff values support non-invasive CT-based assessment but should be interpreted with clinical and laboratory findings; histopathology remains essential for definitive diagnosis. 

Probiotic products are increasingly utilized in animal feed and the food industry to promote health benefits. Herbs such as turmeric (Curcuma longa L.), galingale (Boesenbergia rotunda (Linn.) Mansf.), gotu kola (Centella asiatica (L.) Urb.), and black sesame (Sesamum indicum L.) are rich in bioactive compounds with potential antioxidant, anti-inflammatory, and antimicrobial properties. However, their bioavailability can be limited, and fermentation with probiotics may enhance these attributes. This study investigated the formulation of a synbiotic product by fermenting aqueous extracts of turmeric, galingale, and gotu kola with multi-strain probiotics, supplemented with prebiotic black sesame extract, to improve probiotic viability and biofunctional activities, including enzyme inhibition and antioxidant effects.  Seven probiotic strains (Ligilactobacillus salivarius KUKPS6202, Ligilactobacillus paracasei KUKPS6201, Lactobacillus acidophilus KUKPS6107, Ligilactobacillus rhamnosus KUKPS6007, Ligilactobacillus reuteri KUKPS6103, Bacillus coagulans KPS-TF02, and Saccharomyces cerevisiae subsp. boulardii KUKPS6005) were cultivated in a mung bean–soybean medium and used for fermentation of herbal extracts (10% w/v) at 37°C for 24 h. Black sesame extract (1% w/v) was added as a prebiotic. Antimicrobial activity was assessed via agar well diffusion against pathogens like Aeromonas hydrophila and Bacillus cereus. Probiotic viability was measured by plate counts. Bioactivities evaluated included α-amylase and α-glucosidase inhibition (IC50 values), lipase activity (using a commercial kit), and antioxidant potential via 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging and ferric reducing antioxidant power (FRAP) assays. Data were analyzed using one-way analysis of variance and Tukey's test (p < 0.05).  All probiotic strains grew effectively in the mung bean–soybean medium, with viable counts reaching 7-8 log CFU/mL. Fermented herbal extracts inhibited A. hydrophila and B. cereus (inhibition zones 7.33-9.00 mm) but showed limited activity against other pathogens. Black sesame extract significantly enhanced probiotic viability (p < 0.05) compared to unsupplemented extracts. Fermentation with L. rhamnosus yielded the lowest IC50 for α-amylase (5.96 mg/mL) and α-glucosidase (3.32 mg/mL). L. reuteri exhibited the highest lipase activity (384.77 mU/L after 24 h). Antioxidant activities were comparable across treatments in DPPH assay, but FRAP values were highest for B. coagulans (0.275 μg Trolox/mL), S. boulardii (0.264 μg Trolox/mL), and L. salivarius (0.252 μg Trolox/mL) (p < 0.05).  Synbiotic fermentation with black sesame extract and multi-strain probiotics significantly boosts probiotic survival and biofunctional properties, offering potential as functional foods or animal feed supplements for metabolic health, antimicrobial protection, and antioxidant support. Future in vivo studies could validate these benefits. 

The gut microbiota is essential for nutrient digestion, immune function, and environmental adaptation in ruminants, particularly high-altitude species like yaks (Bos grunniens). Different husbandry practices (captive vs. free-range) can potentially alter the microbial communities and affect the yak health. However, comparative data on how these systems affect yak gut microbiomes remain limited, with most studies focusing on taxonomy rather than functional implications. This study aimed to compare gut microbiome composition, diversity, and predicted functional profiles between captive (CY) and free-range (FY) yaks using a 16S rRNA gene metabarcoding approach.  Fecal samples were collected from healthy ~2-year-old yaks (n=5 CY, n=5 FY) in Litang County, Ganzi Prefecture, Sichuan, China, during summer. DNA was extracted, and the V4 region of the 16S rRNA gene was sequenced on Illumina NovaSeq 6000. Bioinformatic analyses included quality filtering, Operational taxonomic units (OTU) clustering (97% similarity), taxonomic annotation (SILVA database), α- and β-diversity analysis. The microbial function was predicted using PICRUSt2 (KEGG pathways), BugBase (community phenotypes), and FAPROTAX (ecological functions). Statistical comparisan used Welch’s t-tests, Wilcoxon rank-sum tests, principal coordinates analysis (PCoA), and Analysis of similarities (ANOSIM) with significance set at p < 0.05.  α-Diversity indices (e.g., Shannon p = 0.5476) showed no significant differences between CY and FY. However, β-diversity revealed distinct community structures (PCoA: PC1 30.52%, PC2 12.25%; ANOSIM R = 0.976, p = 0.007), with FY samples more homogeneous. At the genus level, CY were enriched in Ruminococcaceae bacterium UCG-005, Streptococcus, Escherichia-Shigella, Treponema, Christensenellaceae R-7, and Clostridium sensu stricto 1 (many fermentative or potentially opportunistic). FY showed higher abundances of Bacillus, Arthrobacter, Rhodococcus, Candidatus Saccharimonas, Prevotellaceae UCG-001, and Paenibacillus. Predicted functions indicated FY had greater capacities for carbohydrate/amino acid metabolism, DNA repair, fatty acid biosynthesis, and vitamin B pathways, while CY favored fermentation and reductive acetogenesis. BugBase highlighted higher anaerobic phenotypes in CY.  Husbandry practices profoundly influence yak gut microbiome structure and inferred metabolic potential, with free-range systems promoting, homogeneous communities suited to natural high-fiber diets while captive systems promotes fermentative and opportunistic shifts. These microbiome differences suggest opportunities for probiotic interventions to enhance yak health, productivity, and sustainability in high-altitude pastoral systems. Future metagenomic and metabolomic validation is needed. 

Epizootic hemorrhagic disease virus (EHDV), an Orbivirus transmitted by Culicoides midges, infects a wide range of ruminant species including dromedary camels (Camelus dromedarius). Although circulation of EHDV has been documented in cattle in the Sultanate of Oman, its epidemiology in camels remains poorly characterized. This study was conducted in the Al Batinah North and Al Batinah South governorates, coastal regions with hot, arid climates favorable to vector activity, to estimate individual-level seroprevalence of EHDV antibodies in dromedary camels, identify associated risk factors using univariate and multivariate analyses, and evaluate spatial distribution patterns to inform future control measures in multi-host livestock systems.  A cross-sectional study was performed between September 2020 and July 2021. Serum samples were collected from 415 camels across 50 holdings using convenience sampling. Antibodies against EHDV were detected with a commercial competitive enzyme-linked immunosorbent assay (c-ELISA) targeting the VP7 protein (sensitivity = 85.9 %, specificity = 99.7 %). True prevalence (TP) was calculated using the Rogan–Gladen estimator. Univariate associations were assessed with chi-square or Fisher’s exact tests; variables with p < 0.25 were entered into backward stepwise binary logistic regression. Spatial analyses included average nearest-neighbour clustering, Getis-Ord Gi* hot-spot analysis, and kernel density estimation in ArcGIS.  Apparent seroprevalence was 40.2 % (95 % confidence interval [CI]: 35.5–45.1 %), with an estimated TP of 46.7 % (95 % CI: 41.3–52.3 %). No significant difference existed between governorates (p = 0.520). In univariate analysis, seropositivity was significantly higher in camels > 4 years of age (45.9 % vs 19.3 %; p < 0.001), in breeding/leisure camels (43.1 % vs 20.8 %; p = 0.002), and in animals kept in contact with other ruminants (49.3 % vs 35.6 %; p = 0.007). After multivariate adjustment, only age > 4 years (odds ratio [OR] = 3.4, 95 % CI: 1.88–5.96; p < 0.01) and contact with other ruminants (OR = 1.6, 95 % CI: 1.05–2.44; p = 0.029) remained independent risk factors. Global spatial clustering of positive holdings was highly significant (nearest-neighbour ratio = 0.39, z = –8.26, p < 0.001), with elevated density along coastal agricultural corridors.  This first large-scale serological survey confirms active circulation of EHDV among dromedary camels in northern Oman. Older age and mixed-species herding are the main risk factors identified. The findings highlight the importance of camels in the multi-host ecology of EHDV and emphasize the need for integrated national surveillance, vector studies, and molecular characterization of circulating serotypes to support evidence-based control strategies in a changing climate.