Palmitic acid (PA) (C16: 0) is a rumen-inert long-chain fatty acid (FA) widely used in dairy cattle to increase dietary energy density and milk fat synthesis; however, its effects in dairy goats remain poorly characterized. This study evaluated whether supplementing 3% or 6% PA in the diet of mid-lactation goats could improve milk yield, composition, FA profile, and whole-animal energy balance under semi-arid Mexican production conditions.  Twenty-one multiparous crossbred goats (45.8 ± 1.2 kg; 21 ± 3 days in milk) were randomly assigned to three treatments for 6 weeks after a 2-week adaptation: (1) Control diet (without PA), (2) diet + 3% PA, and (3) diet + 6% PA on a dry-matter (DM) basis. Diets were isoenergetic and isoproteic before PA addition. Individual DM intake (DMI), milk yield, and composition were measured daily; milk FA profiles and energy balance were determined on days 0, 21, and 42. Data were analyzed using a mixed-model with repeated measures, and means were compared using the Tukey test (p ≤ 0.05).  PA inclusion did not affect DMI, body weight, or milk yield. However, milk fat concentration and yield increased significantly (p < 0.01) in both PA treatments, with the highest fat concentration observed at 6% PA. The milk FA profile shifted toward greater C16: 0 and C16: 1 proportion (p < 0.0001) and decreased short-chain (C16) FA fractions. Energy-corrected milk yield rose by ~40% in PA-fed goats, and energy balance improved markedly from week 3 onward, particularly in the 3% group (p < 0.01), indicating superior dietary energy utilization without intake suppression.  Moderate PA supplementation (~3% DM) effectively enhances milk fat synthesis and energy efficiency in goats while maintaining stable intake and yield. Increasing PA beyond 3% confers minimal additional benefit and may overly saturate milk fat. These findings provide species-specific evidence that rumen-inert fat inclusion can be an efficient strategy to support metabolic status and product quality in mid-lactation goats under variable forage systems. 

Free-range laying hens raised under tropical climates are susceptible to heat stress (HS), which compromises intestinal integrity, nutrient absorption, and egg quality. Ganoderma lucidum (Lingzhi) contains potent antioxidant compounds, but its efficacy in animal diets is limited by poor solubility and stability. This study evaluated the effects of dietary microencapsulated G. lucidum extract (MGE) on laying performance, antioxidant capacity, intestinal morphology, and nutrient digestibility in free-range hens under high-temperature conditions.  A total of 256 Hy-Line Brown hens (25 weeks old) were randomly assigned to four dietary treatments with four replicates of 16 birds each: (i) basal diet (control [Con]), (ii) basal diet + free G. lucidum extract (1, 000 mg/kg), (iii) MGE at 1, 000 mg/kg (MGE0.1), and (iv) MGE at 500 mg/kg (MGE0.05). The trial lasted 12 weeks under natural tropical temperatures. Productive performance, egg quality, fatty acid profile, jejunal histomorphology, and antioxidant indices were analyzed. Statistical analysis was conducted using one-way analysis of variance with Tukey’s test (p ≤ 0.05).  MGE supplementation improved average egg weight and albumen height without affecting feed intake or feed conversion ratio. Both MGE0.1 and MGE0.05 increased jejunal villus height, width, surface area, and the villus height-to-crypt depth ratio compared with the Con. group. MGE diets significantly increased apparent ileal digestibility of dry matter, crude protein, gross energy, and ash. Enhanced antioxidant responses were observed, including higher total antioxidant capacity (15%–19%), superoxide dismutase (15%–22%), and catalase activity, with a concurrent 46%–47% reduction in malondialdehyde. Additionally, MGE diets reduced yolk cholesterol and the n-6/n-3 polyunsaturated fatty acid ratio.  MGE effectively enhances intestinal morphology, antioxidant defense, and nutrient utilization in free-range hens exposed to HS, thereby improving egg quality and the yolk lipid profile. MGE represents a promising natural antioxidant and feed additive for sustainable poultry production under tropical conditions. 

Methicillin-resistant coagulase-positive Staphylococci (MRCoPS), including methicillin-resistant Staphylococcus pseudintermedius (MRSP), methicillin-resistant Staphylococcus aureus (MRSA), and methicillin-resistant Staphylococcus coagulans (MRSC), are emerging zoonotic pathogens in veterinary hospitals, posing significant infection control challenges. This study aimed to investigate the molecular epidemiology, antimicrobial resistance profiles, and clonal dissemination of MRCoPS across environmental surfaces, veterinary personnel, and canine patients at the Veterinary Teaching Hospital, Chulalongkorn University (VTH-CU), Thailand.  A cross-sectional study was conducted involving 216 environmental samples, 23 veterinary staff, and 14 canine patients. Isolates were identified using biochemical tests, polymerase chain reaction (PCR), and matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry. Methicillin resistance was confirmed by mecA gene detection. Antimicrobial susceptibility was evaluated through disk diffusion following Clinical and Laboratory Standards Institute guidelines. Molecular typing was performed using staphylococcal cassette mec (SCCmec) PCR and pulsed-field gel electrophoresis (PFGE). Multivariate logistic regression identified environmental predictors of MRCoPS contamination.  Among 88 coagulase-positive isolates, 62 (70.5%) were methicillin resistant, predominantly MRSP (91.9%), followed by MRSC (8.1%) and MRSA (1.6%). Floors represented the principal environmental reservoir, with significantly higher contamination odds than medical instruments (adjusted odds ratios [AOR] = 0.32; 95% confidence interval = 0.12–0.86; p = 0.024). The dermatological unit showed a six-fold higher risk of MRCoPS contamination than the medicine unit (AOR = 5.86; p = 0.027). All MRSC isolates carried SCCmec type V and displayed a consistent triple resistance pattern (gentamicin-clindamycin-erythromycin), while MRSP isolates exhibited diverse antibiograms and untypeable SCCmec elements. PFGE revealed clonal similarity (pattern A) between canine and environmental isolates, confirming the potential for nosocomial transmission.  MRCoPS, particularly MRSP, were widely distributed and persistent in the VTH-CU environment, despite routine cleaning. The clonal overlap among environmental and canine isolates highlights potential cross-contamination within the hospital. Strengthened disinfection protocols, antimicrobial stewardship programs, and regular environmental surveillance are imperative to mitigate the spread of multidrug-resistant staphylococci. This study highlights the importance of integrating environmental, animal, and human infection control practices in veterinary facilities within the context of One Health. 

Highly pathogenic avian influenza (HPAI) remains a global threat to poultry production, trade, and public health. While Morocco has not yet reported confirmed HPAI outbreaks, the endemic circulation of low-pathogenic avian influenza (LPAI) H9N2 since 2016, proximity to affected neighboring countries, and Morocco’s position along migratory bird flyways highlight the country’s vulnerability. This study aimed to identify high-risk areas for HPAI introduction and spread to inform risk-based surveillance and control policies.  We applied a spatial multi-criteria decision analysis integrated with geographic information systems at the provincial scale. Relevant risk factors were identified through a literature review and expert consultation, and categorized into the introduction (wetlands, live poultry imports, recreational bird imports, and poultry products) and spread (poultry density and type, live bird markets, transport networks, and human population density) domains. Weights were assigned to factors using the analytic hierarchy process based on responses from 73 poultry-sector experts. Data were normalized, integrated into composite risk maps, and validated against historical LPAI H9N2 outbreak data (2016). Sensitivity and uncertainty analyses were used to assess model robustness.  The final maps revealed that 25 provinces (33.3% of the national territory) exhibited high-to-very high risk of HPAI introduction, particularly along northern coastal provinces, border regions, and areas linked to recreational bird imports. For spread risk, 41 provinces (41.3%) were classified as high to very high, concentrated in the Casablanca–Settat, Rabat–Salé–Kenitra, Fès–Meknès, and Marrakech–Safi regions, which are characterized by dense poultry production, major trade hubs, and extensive transport networks. Sensitivity analyses confirmed the model's stability, with variations in weight producing a minimal impact on risk classifications.  This study provides the first comprehensive spatial risk maps of HPAI introduction and spread in Morocco, highlighting priority provinces for early detection, targeted surveillance, and preventive biosecurity measures. Despite limitations arising from reliance on LPAI data and expert judgment, the approach offers a robust decision-support tool for veterinary authorities. The methodology is adaptable to regional applications and can be refined with real-time surveillance data, enhancing Morocco’s preparedness and resilience against future avian influenza incursions. 

An increased proportion of female piglets is desirable in commercial swine breeding to improve productivity, facilitate genetic selection, and reduce the need for male castration. However, currently available sex-selection techniques, such as flow cytometry, are costly and impractical for routine field use. This study evaluated the potential of rabbit serum albumin (RSA) as a low-cost biochemical modulator to influence the proportion of female offspring, comparing its effects with those of other albumin sources and determining optimal supplementation conditions for boar semen used for artificial insemination (AI).  Eight Landrace boars were initially screened in vitro to assess sperm quality and the proportion of X- and Y-bearing sperm following incubation with albumin. Four boars (A, B, E, and G) showing a higher X-sperm proportion were subsequently selected for in vivo trials involving 130 sows. Semen was diluted in a conventional extender supplemented with albumin (RSA, porcine serum albumin, or bovine serum albumin) or left unsupplemented (control). The effects of albumin source, concentration (0.1–0.2 mg/mL), incubation temperature (25°C vs. 37°C), duration (5–15 min), and boar variation were examined. Offspring sex ratio and litter size were analyzed using the Kruskal–Wallis test, followed by Dwass–Steel–Critchlow–Fligner pairwise comparisons (p < 0.05).  All albumin treatments significantly increased (p < 0.05) the proportion of female piglets compared with controls. RSA yielded the greatest effect, particularly at 0.1 mg/mL incubated at 37°C for 15 min, producing up to 61.8% female offspring compared with 24.8% in controls. Boars with an initial male-biased sex ratio showed the largest improvement after RSA treatment. Although litter size decreased slightly with albumin supplementation, the difference was not statistically significant (p ≥ 0.05).  Supplementation of semen extenders with RSA effectively increased the proportion of female piglets without compromising fertility. This method offers a practical, scalable, and economical alternative to conventional sex-sorting technologies for swine breeding. Further optimization and larger-scale validation are warranted to ensure consistent litter performance and broader adoption in commercial production systems. 

Highly pathogenic avian influenza virus (HPAI) H5N1 continues to threaten poultry biosecurity worldwide due to rapid antigenic drift and reassortment. Since late 2020, clade 2.3.4.4b strains have dominated outbreaks across multiple continents. This study genetically characterized H5N1 isolates circulating in Upper Egypt during 2023–2025, clarified their phylogenetic origin, and compared them with vaccine strains used nationally.  A total of 100 samples from 25 broiler flocks showing respiratory and neurological symptoms across New Valley, Assiut, and El-Minya governorates were examined. Specimens were screened for avian influenza subtypes (H5N1, H9N2, H5N8, H6N2) and differential viral pathogens (Newcastle disease virus, infectious bronchitis virus, infectious laryngotracheitis virus, infectious bursal disease virus) using reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Positive isolates were propagated in specific-pathogen-free embryonated chicken eggs and identified through hemagglutination and hemagglutination inhibition assays. Partial hemagglutinin gene sequencing and phylogenetic analyses were performed using Molecular Evolutionary Genetics Analysis version 7.0.  HPAI-H5N1 was detected in 16% (4/25) of flocks, showing 25%–50% mortality. Five isolates displayed high hemagglutination titers (7–8 log2) and were confirmed as H5N1 subtype by RT-qPCR. Phylogenetic analysis classified New Valley-1-H5N1-2023 and New Valley-2-H5N1-2024 within clade 2.3.4.4b. These strains shared 96%–99% nucleotide and amino acid identity with recent Egyptian and Eurasian H5N1 isolates but only 72%–84% with currently used Egyptian vaccine seeds (e.g., MEFLUVAC [Kemin Industries, Inc., USA], EgyFlu [Nagy Awad Group, Cairo, Egypt]). Mutations R72S, A83D, and T140A were identified in receptor-binding and antigenic regions of hemagglutination, implying potential antigenic drift.  This is the first documentation of clade 2.3.4.4b HPAI-H5N1 circulation in broiler flocks of Upper Egypt. The low genetic relatedness to existing vaccine strains indicates probable vaccine mismatch and reduced protection. Continuous molecular surveillance, integration of full-genome sequencing, and periodic vaccine seed updates are essential for effective containment. Enhanced monitoring at the domestic–wild bird interface will help mitigate cross-species transmission and align with One Health strategies for zoonotic risk reduction. 

Accurate evaluation of hormonal status is critical for optimizing growth performance and meat quality in beef cattle. Conventional matrices such as blood, saliva, and urine reflect only short-term fluctuations and are influenced by collection stress. Hair, as a retrospective biosubstrate, can integrate hormone secretion over time and serve as a non-invasive indicator of chronic endocrine activity. This study aimed to evaluate the relationship between hair hormone concentrations and productive performance in Hereford bulls and to establish reference intervals (RIs) for major hormones in hair.  A total of 200 Hereford bulls aged 15–18 months were reared under uniform feeding and housing conditions. Hair samples from the withers were processed into powder (d50 < 20 μm), and concentrations of 12 hormones, including cortisol, adrenaline, testosterone, estradiol, somatotropin (STH), thyroxine (T4), and insulin, were quantified by enzyme-linked immunosorbent assay. Slaughter data included live and carcass weights, yields, and physicochemical meat traits. Correlation analyses (Spearman) and stepwise multiple linear regression were used to determine hormonal predictors of productivity. RIs (2.5th–97.5th percentiles) were calculated according to American Society for Veterinary Clinical Pathology recommendations.  Hair and serum hormones correlated significantly only for STH (r = 0.69) and adrenocorticotropic hormone (r = 0.61). Cortisol and adrenaline were inversely related to anabolic hormones (testosterone and insulin) and showed negative associations with live weight gain, carcass weight, and meat yield, but positive associations with meat pH and lipid oxidation. STH, insulin, estradiol, testosterone, and T4 were positively related to growth rate, carcass composition, and protein content. Regression models identified STH as the strongest independent positive predictor (β = 0.49) and cortisol as the principal negative predictor (β = –0.35) of productivity. RIs for 12 hormones were established for diagnostic application.  Hair hormone analysis reliably reflects chronic endocrine status and predicts productive performance in beef cattle. Elevated stress hormones impair growth and meat quality, whereas anabolic hormones enhance carcass traits. The established RIs can serve as practical benchmarks for metabolic monitoring and herd management strategies in precision beef production. 

Trypanosoma evansi infection (Surra) remains a major constraint to equine health and productivity in Thailand. The only available trypanocidal drug, diminazene aceturate (DA), has limited efficacy, poor blood–brain barrier penetration, and toxicity in horses. This study aimed to investigate the in vitro inhibitory effects of commonly used equine antibiotics, gentamicin (GMC), ceftiofur (CTF), and trimethoprim–sulfamethoxazole (TS), against T. evansi (Thai strain isolated from dairy cattle number 953; TEDC 953) to identify potential therapeutic alternatives or adjuncts for equine trypanosomosis.  An in vitro growth inhibition assay was conducted using the T. evansi TEDC 953 strain cultivated in Hirumi's Modified Iscove's medium 9 (HMI-9 medium) containing 20% horse serum under controlled conditions (37°C, 5% CO2, 75% humidity). Serial dilutions of DA, GMC, CTF, and TS were tested in duplicate across three independent experiments. Parasite viability was assessed after 48 h by microscopic examination, and the half-maximal effective concentration (EC50) was determined using nonlinear regression analysis in GraphPad Prism 5.  Among the three antibiotics, GMC and CTF significantly inhibited T. evansi growth in vitro, whereas TS showed no inhibitory effect. The EC50 values were 1.25 × 10⁻5 ± 3.90 × 10⁻6 mg/mL for DA, 0.22 ± 0.08 mg/mL for GMC, and 0.08 ± 0.05 mg/mL for CTF. Parasite viability assays confirmed that GMC (5 mg/mL) and CTF (0.2 mg/mL) completely eliminated T. evansi after 48 h of exposure. These findings provide the first in vitro evidence of the trypanocidal potential of GMC and CTF against the Thai strain of T. evansi.  GMC and CTF exhibited substantial inhibitory activity against T. evansi under in vitro conditions, supporting their potential use as repurposed or adjunct antibiotics for trypanocidal therapy in horses. This preliminary evidence underscores the need for in vivo validation, pharmacokinetic profiling, and mechanistic studies to explore synergistic effects with conventional trypanocides such as DA. 

Viral infections continue to pose major challenges to pig health, farm productivity, and global food security. Early and accurate diagnosis is the cornerstone of disease prevention, surveillance, and control in swine populations. In recent years, remarkable progress has been achieved in molecular, serological, and digital diagnostic technologies, enabling more rapid, sensitive, and field-adaptable detection of important porcine viruses such as African swine fever virus, porcine reproductive and respiratory syndrome virus, and classical swine fever virus. This review summarizes current and emerging diagnostic approaches, highlighting polymerase chain reaction (PCR) and its advanced forms, quantitative PCR and digital PCR, as the gold standards for laboratory confirmation. The advent of next-generation sequencing and metagenomics has revolutionized pathogen discovery and genomic surveillance, providing comprehensive insights into viral evolution and transboundary transmission. Isothermal amplification techniques such as loop-mediated isothermal amplification and recombinase polymerase amplification have shown strong potential for on-farm diagnosis due to their simplicity, rapidity, and minimal equipment requirements. Innovations such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated-based assays, biosensors, lab-on-a-chip platforms, and point-of-care testing devices are bridging the gap between laboratory precision and field application, allowing rapid decision-making during outbreaks. The integration of artificial intelligence, machine learning, and geographic information systems has further enhanced diagnostic interpretation, real-time data sharing, and early outbreak prediction under the One Health framework. Despite these advances, challenges remain in ensuring assay standardization, affordability, and equitable access in resource-limited regions. Continued international collaboration, data sharing, and policy harmonization under the guidance of the Food and Agriculture Organization, the World Organization for Animal Health, and the World Health Organization are essential for the global control of swine viral diseases. Ultimately, combining molecular innovation with digital adaptability offers the most promising path toward resilient, cost-effective, and sustainable diagnostic systems for safeguarding animal and public health. 

Genetically engineered pigs are invaluable biomedical models for xenotransplantation and the study of human diseases. Although electroporation (EP) and lipofection are individually effective for clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) ribonucleoprotein (RNP) delivery, their combined application in porcine embryos has not been systematically evaluated. This study aimed to determine whether packaging Cas9-guided RNA complexes in cationic lipids enhances EP-mediated gene editing efficiency without compromising embryonic development.  Porcine zygotes with their zona pellucida removed were edited using RNPs targeting beta-1,4-N-acetyl-galactosaminyl transferase 2 (B4GALNT2) or growth hormone receptor (GHR) genes. Four treatment groups were tested: (1) EP with RNPs (EP), (2) EP with lipofectamine-packaged RNPs (EPL), (3) transfection with lipofectamine-packaged RNPs before EP (TL + EPL), and (4) EP followed by lipofection (EPL + TL). Blastocyst formation was evaluated morphologically, and mutation rates were assessed by Sanger sequencing followed by tracking of indels by decomposition (TIDE) analysis.  Blastocyst formation rates were comparable across all treatments, indicating that lipofectamine packaging and EP caused no detectable cytotoxicity. For B4GALNT2, no mutations were induced by EP alone, whereas TL + EPL treatment significantly increased total and mosaic mutation rates (p < 0.05). For GHR, the total mutation and mosaic mutation rates were likewise higher in TL + EPL compared with EP, although mutation efficiency (indel percentage per edited embryo) remained unchanged. These results suggest that pre-EP lipofection promotes RNP uptake by facilitating lipid-membrane interactions that are potentiated by subsequent membrane destabilization through EP.  Packaging RNPs in cationic lipids and applying sequential lipofection followed by EP significantly enhances CRISPR/Cas9-mediated mutagenesis in porcine zygotes without affecting developmental competence. This dual-delivery approach provides a simple, reproducible, and low-toxicity workflow for generating gene-edited embryos, with potential applicability to large-animal biomedical models. 

Ophiotaenia species are globally distributed proteocephalidean cestodes that commonly parasitize amphibians and reptiles. Despite the ecological importance of frogs in controlling insect populations and maintaining food-web stability, data on cestode infections in Egyptian amphibians remain scarce. This study provides the first documentation of Ophiotaenia sp. infecting Amietophrynus kassasii in Egypt and evaluating the in vitro anticestodal activity of Sinularia sp. extract against adult tapeworms.  A total of 85 frogs were collected from freshwater ponds in New Valley Governorate, Egypt, between February and September 2024. Intestines were examined for cestodes, which were identified morphologically using light microscopy and scanning electron microscopy (SEM). Soft coral Sinularia sp. extract was prepared by methanolic extraction, and three concentrations (25, 50, 100 μg/mL) were assessed for anticestodal efficacy using motility, paralysis, and mortality endpoints. Tapeworms from the control and highest-dose groups were subjected to SEM to evaluate tegumental alterations.  Ophiotaenia sp. infection was detected in 5 of 85 frogs (5.9%), with a notably high mean intensity of 70 parasites per host. The recovered cestodes measured 12–30 mm × 0.7–0.9 mm, featuring a scolex with two spherical suckers and a distinct apical organ. Gut-content analysis of infected frogs revealed coleopteran, orthopteran, and hymenopteran insects as probable intermediate or paratenic hosts. Sinularia sp. extract exhibited clear dose-dependent anticestodal activity. Mortality occurred at 7.58 ± 0.15 h (25 μg/mL), 5.79 ± 0.08 h (50 μg/mL), and 4.247 ± 0.09 h (100 μg/mL), compared with 70.39 ± 1.23 h in controls. SEM analysis of treated cestodes showed profound tegumental erosion, sucker shrinkage, cirrus sac constriction, and proglottid contraction, indicating severe structural disruption.  This study documents the first occurrence of Ophiotaenia sp. in A. kassasii in Egypt and provides evidence that Sinularia sp. extract possesses strong, dose-dependent anticestodal properties. The pronounced tegumental damage observed suggests potent cestocidal mechanisms. These findings offer new insights into amphibian parasitology in Egypt and support the potential development of marine-derived natural products as alternative anthelmintics. 

Chitosan-based DNA nanoparticles have emerged as a promising next-generation platform for veterinary vaccines, addressing several limitations of conventional attenuated, inactivated, and recombinant formulations. Chitosan is a biodegradable, biocompatible, and low toxicity polymer with mucoadhesive properties that enhance cellular uptake and protect nucleic acids from enzymatic degradation. These characteristics make it an attractive candidate for delivering plasmid DNA encoding viral antigens across diverse animal species. Recent advances demonstrate that chitosan–DNA nanoparticles can induce robust humoral and cellular immune responses, stimulate mucosal immunity, and achieve high levels of protection in terrestrial livestock, poultry, fish, and crustaceans. A wide range of viral pathogens has been targeted using this approach, including Foot-and-Mouth disease virus, Newcastle disease virus, infectious bronchitis virus, spring viremia of carp virus, white spot syndrome virus, and infectious pancreatic necrosis virus. Depending on the species and formulation strategy, nanoparticles have been successfully administered intranasally, intramuscularly, intraperitoneally, or orally, highlighting their versatility for mass vaccination in both terrestrial and aquatic systems. Reported protection rates range from 60% to 100% in mammalian and avian models, while oral nanoparticle vaccines in shrimp and fish have demonstrated sustained immune activation and survival benefits. The ability to incorporate genetic adjuvants, such as cytosine-phosphate-guanine motifs, cytokines, or complement fragments, further enhances the immunogenicity of these platforms. Despite these promising results, several challenges remain. Most studies use small laboratory animals or controlled experimental settings, and data from large-scale field trials in cattle, pigs, and equines remain scarce. The stability of nanoparticle formulations during long-term storage, the scalability of manufacturing processes, and the standardization of dosing regimens require further investigation. Overall, chitosan–DNA nanoparticles represent a safe, flexible, and rapidly adaptable vaccine carrier system with significant potential to transform veterinary immunization. Their capacity to elicit mucosal and systemic immunity, enable needle-free delivery, and support DIVA-compatible vaccine design positions them as a valuable tool for controlling emerging and re-emerging viral diseases in the context of One Health. 

Listeria monocytogenes remains a major foodborne pathogen globally, with mortality rates ranging from 20%–40%. The increasing incidence of listeriosis and the limitations of culture-based and polymerase chain reaction-based diagnostics highlight the need for rapid, cost-effective, and highly specific immunoassays. This study aimed to develop and validate a regionally adapted sandwich enzyme-linked immunosorbent assay (ELISA) based on monoclonal and polyclonal antibodies (pAbs) raised against a recombinant p60 antigen derived from a Kazakhstani L. monocytogenes field isolate.  The p60 gene lacking its N-terminal signal peptide was amplified from a regional L. monocytogenes isolate, cloned into the pET28c(+) vector, and expressed in Escherichia coli arctic express (DE3). Recombinant p60 protein was purified by Ni2⁺-affinity chromatography and used to immunize BALB/c mice and Chinchilla rabbits for monoclonal antibodies and pAbs antibody production. Hybridoma clones were screened for specificity using indirect ELISA and Western blot. A sandwich ELISA was assembled using mAb 1H8 as the capture antibody and horseradish peroxidase-conjugated rabbit pAbs as detection antibodies. Analytical sensitivity and diagnostic performance were evaluated using serial dilutions of recombinant p60 and culture supernatants of L. monocytogenes isolates recovered from 507 food samples.  The recombinant p60 antigen (50.3 kDa) was successfully expressed and purified at 5.9 mg/L yield. Among seven stable hybridoma clones, mAb 1H8 exhibited the highest affinity (Ka = 2.5 × 1010 M⁻1) and specificity without cross-reactivity to non-Listeria bacteria. The optimized sandwich ELISA achieved a detection limit of 1.5 ng/mL, corresponding to approximately 103 colony-forming units/mL. All six L. monocytogenes field isolates tested positive in the assay, with results strongly correlating with viable cell counts (R2 = 0.89). The assay demonstrated comparable sensitivity to commercial kits while offering shorter assay time (2 h) and substantially lower production cost.  The developed sandwich ELISA provides a sensitive, specific, rapid, and regionally tailored diagnostic platform for detecting pathogenic L. monocytogenes in food samples. By integrating locally produced recombinant antigens and immunoreagents, the assay offers a cost-effective alternative to imported kits, supporting national food safety programs and One Health surveillance initiatives. 

Enteric methane (CH₄) emissions from ruminants reduce dietary energy efficiency and contribute to global greenhouse gas accumulation. Gliricidia sepium (Gamal), a protein-rich tropical legume, is widely used as a basal forage but may require targeted additives to optimize rumen fermentation. Plant bioactive compounds, such as Uncaria gambir (Gambier), and microbial supplements, such as direct-fed microbials (DFM) containing Saccharomyces cerevisiae, are known to influence fermentation pathways and energy use. This study aimed to compare the effects of Gamal-based diets supplemented with Gambier or DFMs on nutrient digestibility, rumen fermentation characteristics, volatile fatty acids (VFAs), microbial protein synthesis (MPS), fatty acid (FA) profiles, and CH₄ mitigation under in vitro conditions.  Six dietary treatments were evaluated in a randomized block design with three replicates: control, Gamal substitution, Gamal with 0.5% DFM, Gamal with 1% DFM, Gamal with 1% Gambier, and Gamal with 2% Gambier. Substrates were incubated in rumen fluid and buffer for 48 h at 39°C. Dry matter (DM) digestibility, organic matter digestibility, ammonia-nitrogen (NH3-N), VFAs, CH₄ production, MPS, and FA composition were analyzed using standard procedures. Statistical analysis was performed using analysis of variance followed by Duncan’s test.  Supplementation with DFM improved DM and organic matter digestibility, with the highest values observed in the diet containing 0.5% DFM. The greatest methane reduction was observed with 1% Gambier, which lowered CH₄ output by 33.43% compared with the control. DFM increased total VFAs and propionate concentration, reducing the acetate-to-propionate ratio. Gambier increased the acetate concentration and NH3-N and achieved the highest MPS values. FA profiles shifted according to additive type, with notable changes in saturated, monounsaturated, and polyunsaturated FA.  Gamal-based diets supplemented with Gambier or DFM positively modified rumen fermentation, but through distinct mechanisms. DFM improved digestibility and fermentation stability, whereas Gambier achieved the greatest CH₄ mitigation at 1% inclusion. Gambier represents a promising, locally available option for sustainable methane reduction in tropical ruminant feeding systems, supporting future in vivo validation. 

Biofilm-forming Aeromonas hydrophila represents a critical constraint in aquaculture, driving recurrent infections, environmental persistence, and antimicrobial resistance. Sustainable alternatives to antibiotics are urgently needed. This study evaluated the multiphase antibiofilm activity of chitosan nanoparticles (ChNPs) synthesized from Litopenaeus vannamei shrimp shells against clinical A. hydrophila isolates from Indonesian gourami (Osphronemus gouramy), focusing on their effects during biofilm adhesion, planktonic proliferation, and mature biofilm degradation.  Between February 2024 and March 2025, diseased gourami were sampled from aquaculture sites in Surabaya, Indonesia. Three wild-type A. hydrophila isolates (A1G1, A2G1, A3G1) were confirmed via biochemical and 16S rRNA sequencing. ChNPs were synthesized through ionic gelation of deacetylated chitosan with sodium tripolyphosphate and characterized by Scanning Electron Microscopy (SEM), dynamic light scattering, and Fourier Transform Infrared Spectroscopy (FTIR) analyses. Antibiofilm efficacy was tested at concentrations of 15–45 μg mL⁻¹ using crystal violet staining (optical density [OD]₅₉₅) for adhesion and degradation phases, and turbidity (OD₆₀₀) for planktonic inhibition. Data were analyzed using one- and two-way analysis of variance with Tukey’s post hoc test.  ChNPs exhibited spherical morphology (≈641 nm; ζ = +51 mV) and stable ionic crosslinking. They significantly inhibited adherent biomass formation (p < 0.05), reducing OD₅₉₅ from 0.787 to 0.317 in the most responsive strain A3G1 (> 59 % inhibition). Planktonic growth (OD₆₀₀) declined dose-dependently (63 % inhibition at 45 μg mL⁻¹), with significant strain–concentration interactions (p < 0.01). Mature biofilm degradation reached 63% at 45 μg mL⁻¹, approaching the level of the antibiotic-treated control. SEM and FTIR data supported electrostatic disruption and extracellular polymeric substance penetration as probable mechanisms.  Shrimp-shell–derived ChNPs effectively suppressed A. hydrophila biofilms at multiple developmental stages, demonstrating a potent, biodegradable alternative for the control of aquaculture pathogens. Their integration into eco-friendly, antibiotic-free disease management aligns with circular bioeconomy and One Health frameworks. Further in vivo validation and formulation optimization are warranted. 

Feline coronavirus (FeCoV) is a widely circulating Alphacoronavirus that causes mild enteric infections and, in some cases, progresses to Feline infectious peritonitis, a fatal systemic disease. FeCoV consists of two genotypes (I and II) and two biotypes (FeCoV and feline infectious peritonitis virus [FIPV]). Despite its importance, whole-genome data, particularly for FeCoV genotype II, remain limited in Thailand. This study aimed to determine the prevalence of FeCoV in domestic cats and to genetically characterize circulating strains using whole-genome and S gene sequencing.  A total of 471 rectal swabs were collected from domestic cats presented to private small animal hospitals in Bangkok and neighboring provinces from October 2022 to October 2023. FeCoV detection and genotyping were performed using one-step reverse transcription polymerase chain reaction targeting the 3′UTR and S gene, respectively. Selected FeCoV-positive samples were subjected to whole-genome sequencing (WGS) (n = 4) and complete S gene sequencing (n = 6) using Oxford Nanopore technology with Minimap2, Racon, and Medaka pipelines. Phylogenetic and genetic analyses were conducted using MEGA program.  FeCoV positivity was 21.87% (103/471), with higher detection in young cats (<6 months; 28.46%), though age, clinical status, and season showed no significant association (p > 0.05). Genotype I was overwhelmingly predominant (99.03%), whereas genotype II was rare (0.97%). Phylogenetic analysis revealed that Thai FeCoV-I strains clustered closely with Chinese and Dutch FeCoV-I strains, while the FeCoV-II strain grouped with Chinese FeCoV-II. Whole-genome pairwise comparisons showed high nucleotide and amino acid identities with their respective genotype references. No mutations were detected in the S1/S2 or S2 cleavage sites of Thai FeCoV-I, indicating conserved spike characteristics typical of FECoV biotypes. FeCoV-II exhibited the characteristic deletion and insertion patterns known for this genotype. No evidence of recombination with other coronaviruses was observed.  This study provides updated molecular epidemiology of FeCoV in Thailand and reports the first complete FeCoV-II genome sequences from the country. The predominance of FeCoV-I and the detection of conserved spike regions highlight the need for genotype-specific surveillance and the reconsideration of vaccine strategies that currently target FeCoV-II. Expanded nationwide monitoring and detailed recombination analyses are warranted to better understand FeCoV evolution and transmission in feline populations. 

Plastic-derived monomers such as styrene are increasingly detected in tropical freshwater ecosystems at concentrations approaching 0.8 mg/L. These contaminants pose toxicological risks to aquatic organisms, particularly through oxidative stress, inflammation, and metabolic disruption. Oryzias celebensis (Celebes medaka), an endemic tropical freshwater species, offers high ecological relevance for assessing pollutant impacts in Southeast Asia. This study evaluated the hepatic histopathological and molecular responses of O. celebensis following acute styrene exposure and assessed its suitability as a tropical sentinel species.  Adult O. celebensis were exposed to 0.1, 0.25, 0.5, and 0.75 mg/L styrene for 96 h under semi-static conditions. Liver tissues were examined for cytoplasmic vacuolization, necrosis, inflammatory cell infiltration, and congestion using standard histopathological scoring. Expression of biomarker genes related to detoxification (cytochrome P450 1A1 [CYP1A1]), oxidative stress (catalase, superoxide dismutase [SOD]), and inflammation (tumor necrosis factor [TNF]) was quantified using reverse transcription quantitative polymerase chain reaction. Correlations between biomarker expression and tissue lesions were analyzed using Spearman’s coefficients.  Styrene exposure induced concentration-dependent hepatic injury. Histopathological lesions intensified markedly at ≥0.5 mg/L, with prominent vacuolization, necrosis, and inflammatory infiltration. CYP1A1 was strongly upregulated, showing a 36.9-fold increase at 0.75 mg/L (p < 0.01), indicating robust activation of the aryl hydrocarbon receptor (AhR) pathway. Antioxidant enzymes (catalase and SOD) and TNF expression also increased significantly, reflecting oxidative and inflammatory stress. Strong positive correlations (rₛ = 0.93–0.99) were observed between gene expression and lesion severity, confirming mechanistic links between molecular responses and tissue pathology.  Acute styrene exposure triggers coordinated hepatocellular injury and molecular stress responses in O. celebensis through activation of aryl hydrocarbon receptor-CYP1A1, nuclear factor erythroid 2-related factor 2, and nuclear factor kappa-light-chain-enhancer of activated B cells pathways. The strong correspondence between histopathological and transcriptional biomarkers demonstrates that this species is highly sensitive to styrene toxicity and suitable for ecotoxicological monitoring. Findings highlight the need for environmental surveillance of industrial pollutants in tropical freshwater systems. 

Feline chronic enteropathy (CE), often manifesting along the triaditis-axis with concurrent pancreatitis, remains difficult to manage despite standardized dietary modification and cobalamin supplementation. Dysregulated macrophage activity contributes to persistent mucosal and pancreatic inflammation. EF-M2 (ImmutalonTM, Activator MAF LLC, Russia) is an analytically defined, alpha-N-acetylgalactosamine (α-GalNAc) –bearing Gc protein-derived macrophage-activating factor 2.0 (GcMAF 2.0) ligand designed to engage C-type lectin domain family 10 member A (CLEC10A) and promote M2-leaning macrophage-programming. This study aimed to evaluate whether adjunct EF-M2 improves clinical disease activity compared with placebo and to determine whether clinical responses align with macrophage-linked pharmacodynamic (PD) biomarkers.  A multicenter, randomized, double-blind, placebo-controlled, parallel-group trial was conducted in client-owned cats with CE (modified intention-to-treat = 36). Cats received EF-M2 or volume-matched saline twice weekly for 4 weeks in addition to standardized diet/B12 care, followed by a 4-week off-drug period (day 56). The primary endpoint was the change in the feline CE activity index (FCEAI) at day 28. Secondary outcomes included responder rate (≥50% reduction), steroid-sparing effect, serum specific feline pancreatic lipase (Spec fPL), blinded abdominal ultrasonography, PD markers arginase-1 to inducible nitric oxide synthase (ARG1/iNOS) ratio, interleukin-10 [IL-10], and tumor necrosis factor-alpha [TNF-α]). Safety was assessed using Veterinary Cooperative Oncology Group – Common Terminology Criteria for Adverse Events (VCOG-CTCAE) criteria.  EF-M2 significantly improved FCEAI scores at day 28 compared with placebo (least-squares mean difference −2.5; 95% confidence interval −3.7 to −1.3; p = 0.0007). Responder rates were higher with EF-M2 (61% vs. 28%), and more cats remained steroid-free through day 28 (72% vs. 39%). Clinical benefits partially persisted to day 56 (between-group difference in FCEAI −2.1; p = 0.004). In the pancreatitis-positive subgroup, EF-M2 produced a greater reduction in Spec fPL (−2.1 vs. −0.3 μg/L; p = 0.009) and improved pancreatic ultrasonography indices. PD markers shifted consistently with the intended mechanism (ARG1/iNOS ↑, IL-10 ↑, TNF-α ↓; all p ≤ 0.01), and ΔARG1/iNOS correlated with ΔFCEAI (r = −0.57; p = 0.001). Adverse events were mild and comparable between groups, with no treatment-related serious events.  Short-course adjunct EF-M2 achieved clinically meaningful improvement in disease activity, reduced steroid exposure, and improved pancreatitis-associated indicators in cats with CE. The coherent M2-leaning PD signature supports macrophage-programming as a biologically plausible mechanism. EF-M2 demonstrated favorable tolerability and represents a promising adjunctive option for triaditis-axis disease. 

Non-starch polysaccharides (NSP) in cereal-based poultry diets can impair nutrient digestibility and increase intestinal viscosity, reducing growth performance. Supplementation with NSP-degrading enzymes such as endo-1,4-xylanase and β-glucanase can improve energy utilization and feed efficiency. This study evaluated the effects of xylanase alone or in combination with β-glucanase on growth performance, carcass yield, and meat quality of broiler chickens fed energy-reduced diets.  A total of 320 male Ross 308 broilers were distributed into four treatments with eight replicates of ten birds: Positive control (PC), negative control (NC; −100 kcal dietary metabolizable energy/kg), NC + xylanase (12.45 Internation Units [IU]/kg), and NC + xylanase + β-glucanase (12.45 + 12.8 IU/kg). Birds were reared for 37 days under a three-phase feeding program (starter, 1–10 days; grower, 11–24 days; finisher, 25–37 days). Growth indices, feed conversion ratio (FCR), energy conversion ratio (ECR), and European production efficiency factor (EPEF) were recorded. At 38 days, carcass yield, organ weights, and breast meat quality (pH, color, drip loss, wooden-breast, and white-striping scores) were assessed.  During the grower phase, enzyme supplementation significantly improved FCR, ECR, and EPEF compared with NC (p < 0.05). The combined xylanase + β-glucanase treatment produced energy-efficiency and cost benefits comparable to PC. In the finisher phase, xylanase alone improved FCR and ECR (p < 0.05). No significant effects on carcass yield, internal organ proportions, or breast meat quality were observed (p > 0.05). The incidence of wooden-breast and white-striping myopathies remained minimal across all groups.  Supplementation with xylanase and β-glucanase enhances nutrient utilization and feed efficiency in low-energy corn–soybean diets without compromising carcass or meat quality. A combination of xylanase + β-glucanase is most beneficial during the grower phase, while xylanase alone optimizes performance during the finisher phase, providing a cost-effective and sustainable feeding strategy for broiler production. 

Ticks and tick-borne diseases (TBDs) remain a major constraint to cattle production, responsible for substantial economic losses through reduced productivity, increased treatment costs, and high mortality. Beyond livestock impacts, ticks transmit a range of zoonotic pathogens, posing significant health risks to communities in close contact with cattle. Despite Borno State having the largest cattle population in Nigeria, there is no prior documentation of cattle farmers’ knowledge, attitudes, and practices (KAP) regarding ticks, TBDs, and their zoonotic implications. This study aimed to assess farmers’ awareness, preventive behaviors, and tick-control strategies, while evaluating the influence of formal and informal education on these variables.  A cross-sectional descriptive KAP survey was conducted among 492 cattle farmers across Maiduguri Metropolitan Council and Jere Local Government Area between November 2024 and February 2025. Data were collected using a semi-structured and pre-validated questionnaire translated into local languages. Descriptive statistics summarized KAP, while Chi-square tests assessed associations between education and key outcome variables (significance level: p ≤ 0.05).  Most farmers (77.2%) reported observing ticks on their cattle, and 82.9% recognized their role in livestock disease transmission. Tick occurrence was highest during the rainy season (83.7%). Although awareness of livestock TBDs was high, more than half (54.4%) were unaware that ticks transmit diseases to humans. A large proportion (59.8%) reported previous tick bites, but only 10.2% sought medical care afterward. Combined control through acaricides and handpicking was the predominant practice (78.9%). Significant differences between formally and informally educated farmers were observed for lesion recognition after tick bites (χ² = 128.04; p ≤ 0.001), tick-control method (χ² = 26.30; p ≤ 0.001), frequency of handpicking (χ² = 44.27; p ≤ 0.001), and acaricide application methods (χ² = 57.45; p ≤ 0.001).  Farmers demonstrated good knowledge of ticks and livestock TBDs but exhibited low awareness of zoonotic risks and poor health-seeking behavior following tick bites. Strengthening public health education, promoting protective practices, and integrating zoonotic TBDs into One Health policies are essential to reducing risks among high-exposure populations. 

The Ovar-DRB1 gene, a key component of the sheep main histocompatibility complex (MHC) class II region, plays a critical role in antigen presentation and immune responsiveness. Despite the well-documented hypervariability of exon 2 in many sheep breeds, no study has yet examined DRB1 allelic composition in Edilbay sheep, a Kazakh breed highly adapted to harsh continental steppe conditions. This study aimed to characterize the diversity of Ovar-DRB1 exon 2 alleles in Edilbay sheep and to identify novel allelic variants using Sanger sequencing.  Blood samples from 50 Edilbay sheep reared at a breeding farm in Kazakhstan were subjected to DNA extraction and DRB1 exon 2 amplification using validated primers. Polymerase chain reaction products were purified and sequenced using Sanger sequencing. Allele identification was performed through pairwise sequence alignment in SnapGene and reference comparison with the Immuno Polymorphism Database of Major Histocompatibility Complex (IPD-MHC) database. Ambiguous chromatograms and overlapping nucleotide peaks were assessed for potential novel allelic patterns. Genetic diversity indices (Ho, He, Ne, and Shannon’s H’) were calculated.  A total of 25 known DRB1 alleles were identified in the Edilbay sheep population. Genetic diversity parameters demonstrated extremely high immunogenetic variation, with observed heterozygosity (Ho) of 0.94, expected heterozygosity (He) of 0.90, an effective number of alleles (Ne) of 16.7, and Shannon’s index (H’) of 3. Several chromatograms showed overlapping peaks or substitution patterns inconsistent with known alleles, including variations at positions 243–244 and multiple additional polymorphic sites. These patterns indicate the presence of putative novel alleles that could not be unambiguously assigned by direct Sanger sequencing. Approximately 20% of samples contained undocumented variants or low-quality chromatograms requiring further resolution.  This study presents the first comprehensive molecular characterization of Ovar-DRB1 exon 2 in Edilbay sheep, revealing exceptionally high genetic diversity and strong evidence for previously undescribed alleles. These findings broaden the catalog of DRB1 variants and highlight the breed’s adaptive immunogenetic potential. Further investigations using allele-specific amplification, cloning, or next-generation sequencing are recommended to precisely identify novel variants and explore associations with disease resistance and environmental adaptation. 

Canine monocytic ehrlichiosis, caused by Ehrlichia canis and transmitted primarily by Rhipicephalus sanguineus, is a common yet diagnostically challenging tick-borne disease in tropical regions. On the northern coast of Perú, environmental conditions favor vector persistence, but local data on clinical characteristics and risk determinants remain limited. This study aimed to determine the seroprevalence of E. canis in domestic dogs in Trujillo (La Libertad, Perú), describe associated clinical findings, and identify epidemiological risk factors linked to infection.  A cross-sectional analytical study was conducted from December 2023 to August 2024 involving 462 dogs with compatible clinical signs and/or tick infestation from 18 veterinary clinics across three districts. Serological testing was performed with the CaniV-4® rapid test, and hematological parameters were analyzed with an automated analyzer. Epidemiological data were obtained through owner questionnaires. Associations were evaluated using chi-square tests, logistic regression (Odds ratio [OR], 95% CI), and Mann–Whitney U tests for hematological differences. A p-value < 0.05 with OR and lower CI >1 defined risk factors.  The overall seroprevalence of E. canis was 51.3% (95% CI: 46.7%–55.8%). Sex and breed were not associated with infection. Dogs <1 year old (OR = 1.46), those lacking external deworming (OR = 1.99), fed homemade diets (OR = 2.26), and those frequently contacting stray dogs (OR = 4.33) were at significantly higher risk. Clinical predictors strongly associated with infection included lethargy (OR = 5.55), fever (OR = 5.52), anorexia (OR = 4.24), anemia (OR = 4.12), lymphadenopathy (OR = 3.46), and epistaxis (OR = 2.50). Seropositive dogs exhibited significantly reduced erythrocyte counts, hematocrit, hemoglobin, leukocyte counts, and platelet counts (p < 0.01). Although tick presence and park access were associated with seropositivity, their OR < 1 suggested confounding rather than true protective effects.  The high seroprevalence and significant clinical–hematological alterations highlight widespread exposure to E. canis among dogs in northern coastal Perú. Identified risk factors emphasize the need for integrated tick-control, improved owner awareness, and strengthened diagnostic protocols. Future research combining molecular confirmation, socioeconomic variables, and One Health–based surveillance is recommended to refine prevention and management strategies. 

The rapid rise of antimicrobial resistance threatens effective infection control and reinforces the need for alternative therapeutics. Chromolaena odorata (Siam weed), a traditionally used medicinal plant rich in phenolic and flavonoid compounds, has been reported to possess antimicrobial properties. This study evaluated the antimicrobial, antibiofilm, antioxidant, and ultrastructural effects of ethanolic C. odorata leaf extract against a diverse panel of Gram-positive and Gram-negative bacteria.  Ethanolic crude extract was prepared from dried C. odorata leaves, and its antimicrobial activity was assessed against 46 bacterial isolates using disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. Total phenolic and flavonoid contents were quantified using Folin–Ciocalteu and aluminum chloride methods. Antioxidant activity was measured using the 2,2-diphenyl picrylhydrazyl (DPPH) radical scavenging assay. Antibiofilm efficacy against Bacillus cereus was determined using crystal violet staining at sub-MIC levels. Ultrastructural alterations in B. cereus were examined via scanning electron microscopy (SEM).  The crude extract inhibited 78.26% (36/46) of tested isolates, with strong activity against nine species, including B. cereus, Staphylococcus aureus, Staphylococcus epidermidis, Micrococcus luteus, Aeromonas hydrophila, Stenotrophomonas maltophilia, Citrobacter freundii, and Shigella sonnei. MIC values ranged from 31.25–125 mg/mL, with B. cereus showing the lowest MIC and MBC (31.25 mg/mL). The extract exhibited high phenolic (96.82 ± 2.07 μg Gallat-equivalents/mg) and flavonoid (62.98 ± 2.64 μg Quercetin equivalent /mg) content, and moderate antioxidant activity (IC₅₀ = 120.02 ± 16.31 μg/mL). Sub-MIC concentrations significantly inhibited B. cereus biofilm formation in a dose- and time-dependent manner, achieving up to 66.16% inhibition at 1/2 MIC after 72 h (p < 0.001). SEM analysis revealed cell shrinkage, wall collapse, and surface roughening in treated B. cereus, indicating disrupted cell integrity.  Ethanolic C. odorata extract demonstrates broad-spectrum antibacterial, antibiofilm, antioxidant, and cell-disruptive activities, with pronounced effects against B. cereus. These findings highlight its potential as a natural antimicrobial or disinfectant candidate and support future development of plant-based agents to mitigate resistant bacterial infections. 

Gastrointestinal (GI) parasites significantly affect cattle productivity and animal health, especially in tropical regions where environmental and management conditions favor parasite survival. In Mozambique, most previous studies have focused on goats or individual parasite species, leaving crucial gaps in understanding multi-parasite burdens in cattle. This study aimed to determine the prevalence, diversity, and risk factors associated with GI parasites in cattle from the Limpopo district of southern Mozambique, applying a One Health lens due to the zoonotic potential of some parasites that circulate in cattle.  A cross-sectional study was conducted from January to May 2025 using 200 stool samples collected directly from cattle rectums. Samples were examined using Ritchie centrifugal sedimentation for helminths and protozoa and Ziehl–Neelsen staining for Cryptosporidium spp. Epidemiological data on grazing areas, deworming practices, and animal demographics were collected to identify risk and protective factors through Fisher’s test and odds ratios (OR).  Overall, 88.5% of cattle harbored at least one GI parasite. Eight parasite groups were detected: Eimeria spp. (49%), Strongyle-type eggs (46.5%), ciliates (29.5%), Paramphistomum spp. (18%), Fasciola spp. (11%), Cryptosporidium spp. (3.5%), Giardia spp. (2.5%), and Entamoeba spp. (1.5%). Grazing in non-flooded areas significantly reduced Fasciola spp. infection. (OR = 0.126) and Paramphistomum spp. (OR = 0.236), whereas deworming reduced Strongyle-type infections (OR = 0.366). Conversely, dewormed animals had higher odds of Eimeria spp. and ciliate infections, likely because ivermectin was ineffective against protozoa. Co-infections were common, particularly among adult animals.  This first multi-parasite epidemiological assessment in Mozambican cattle reveals a high burden of GI parasites, influenced by grazing conditions and suboptimal deworming practices. Avoiding flooded grazing areas, adopting coccidiostats, and implementing anthelmintic rotation are crucial for effective parasite control. Given the zoonotic relevance of Cryptosporidium, Giardia, and Fasciola, molecular studies are urgently needed to characterize circulating species and clarify the role of cattle as reservoirs. These findings provide essential evidence to strengthen veterinary surveillance and inform One Health interventions in southern Mozambique. 

Sperm sex-sorting technologies are crucial tools for enhancing reproductive management, production efficiency, sustainability, and animal welfare. Although widely used across various species, the effectiveness of these technologies, particularly between flow cytometry and emerging alternative methods, remains inconsistently reported. This systematic review and meta-analysis examined research published worldwide over the past 20 years to compare the performance of different sperm sex-sorting techniques, evaluate pregnancy outcomes and offspring sex ratios, and identify research trends, limitations, and knowledge gaps.  A thorough search of PubMed, Scopus, and Google Scholar identified studies published from 2005 to 2025. Ninety-one studies met the criteria for systematic review, and 22 were included in the quantitative synthesis. Data extraction included species, sex-sorting method, experimental design, sperm quality parameters, pregnancy outcomes, and female-offspring proportions. Random-effects proportional meta-analyses were conducted for pregnancy rate and sex-ratio outcomes. Heterogeneity was assessed using I², Tau², and H², while publication bias was evaluated with funnel plots and Egger’s/Begg’s tests.  Flow cytometry was the most widely used method (64 studies), especially in North America, while research in Asia focused more on alternative techniques such as antibody-based sorting, centrifugation, modified buffers, magnetic beads, and microfluidics. Meta-analysis of in vivo studies showed an overall pregnancy rate of 46.47% for flow cytometry, with significant differences among cattle, buffalo, and deer. Centrifugation-based methods had a higher pooled pregnancy rate of 66.49%, though this was based on only three studies. The overall proportion of female offspring after flow-sorting across all species was 81.72%, with cattle reaching 84.40%. Significant heterogeneity was present (I² > 73%), but no publication bias was found.  Flow cytometry remains the global gold standard for sperm sex-sorting, offering high accuracy, but it is limited by cost, technical requirements, and reduced accessibility in developing regions. Growing research activity in Asia highlights increasing interest in alternative, low-cost methods; however, in vivo validation remains limited. Future efforts should focus on large-scale field trials, standardized protocols, comparative studies with unsorted semen, and expanded research across species, including wildlife, to improve practical applicability, sustainability, and One Health benefits. 

Aeromonas hydrophila is a zoonotic, antimicrobial-resistant pathogen that causes significant losses in aquaculture and raises important One Health concerns. Outer membrane protein (OMP)–based subunit vaccines provide a targeted, antibiotic-sparing alternative to traditional bacterins, but validation across mammalian species remains limited. This study assessed the immunogenicity, safety, and protective effectiveness of a native ~34 kDa Omp34 (nOmp34) subunit vaccine in BALB/c mice, comparing it to a formalin-killed cell (FKC) vaccine, and examined immune factors that may predict survival.  Female BALB/c mice (n = 13 per group) received subcutaneous injections of phosphate-buffered saline (PBS), FKC, FKC + incomplete Freund’s adjuvant (IFA), or native Omp34 + IFA on days 0, 14, and 28. Immune responses were assessed by measuring anti-Omp34 immunoglobulin (Ig)G2a levels via enzyme-linked immunosorbent assay, serum lysozyme activity, nitroblue tetrazolium (NBT) respiratory burst, and phagocytic activity at specified intervals up to day 42. On day 42, mice were challenged intraperitoneally with a lethal dose of A. hydrophila, causing 80% mortality, and observed for 14 days for survival, clinical scores, and body weight changes. Data analysis involved analysis of variance with Tukey post hoc tests, mixed-effects modeling, Spearman correlation, receiver operating characteristic curves, logistic regression, and Kaplan–Meier survival analysis.  By day 42, all immune biomarkers showed clear separation (nOmp34+IFA > FKC + IFA > PBS; p < 0.05). NBT demonstrated the strongest correlation with survival (ρ ≈ 0.90) and the highest predictive performance (Area under the curve [AUC] ≈ 0.80), exceeding IgG2a and phagocytosis (AUC ≈ 0.70). Post-challenge survival rates were 84.6% for nOmp34 + IFA, 61.5% for FKC + IFA, and 23.1% for PBS, corresponding to relative percent survival values of 80% and 50% compared to PBS. The direct comparison between nOmp34 and FKC revealed a favorable but not statistically significant survival benefit (p = 0.238). Vaccination was well-tolerated, with stable body weight, minimal reactogenicity, and no severe clinical events.  The nOmp34 subunit vaccine elicited a strong, coordinated humoral and innate immune response, surpassing the matched bacterin in both efficacy and immune strength. NBT activity between days 35–42 proved to be a practical indicator of protection, aligning mechanistically with nicotinamide adenine dinucleotide phosphate -oxidase–mediated bacterial killing. These findings offer proof-of-concept for Omp34 as a scalable, antibiotic-sparing vaccine candidate and support its progression into aquaculture-relevant platforms within a One Health framework. 

Hypertrophic cardiomyopathy (HCM) is the most common form of cardiomyopathy in cats and is often linked with diastolic dysfunction and progressive remodeling of the left atrium (LA). While LA strain analysis has been utilized to measure atrial function, LA stiffness (LASt), a load-adjusted marker that combines diastolic filling pressures and atrial deformation (E:E’/εS), has not yet been studied in feline patients. This study aimed to compare LASt between normal cats and those with HCM and to assess its correlation with traditional echocardiographic parameters of LA and left ventricular (LV) function.  This retrospective cross-sectional study included client-owned cats evaluated at a university teaching hospital between August 2021 and August 2022. Cats were classified as normal or HCM based on LV wall thickness (≤5 mm vs. ≥6 mm). Standard echocardiographic parameters, doppler indices, tissue Doppler imaging (TDI) values, and two-dimensional speckle-tracking echocardiography (STE)–derived LA strain variables (reservoir, conduit, and active strain) were measured. LASt was calculated as E:E’/εS. Group comparisons were performed using t-tests or Mann–Whitney U tests, and correlations were assessed using Pearson’s coefficient.  Thirty-seven cats met the inclusion criteria (12 with HCM and 25 normal). Cats with HCM showed significantly higher LASt values compared to normal cats (median 2.26 vs. 0.30; p < 0.001), representing approximately a sevenfold increase. Reservoir strain (εS), conduit strain (εE), and active strain (εA) were all significantly lower in the HCM group (p < 0.001). LASt showed strong positive correlations with LA diameter and LA:Ao ratio (r ≥ 0.85; p < 0.001), and moderate-to-strong correlations with Doppler and TDI markers of diastolic dysfunction, including E:A ratio, MV A velocity, MV E’, and MV S’.  LASt, measured by STE, provides a sensitive, load-inclusive index of LA mechanical dysfunction in cats with HCM and may detect atrial remodeling earlier than strain alone. Its strong association with established markers of diastolic impairment supports its potential clinical utility for identifying subclinical atrial dysfunction. Larger longitudinal studies are warranted to validate LASt as a prognostic biomarker and to define clinically relevant thresholds for disease staging and monitoring. 

The inland aquaculture sector in Saudi Arabia has expanded rapidly, with Nile tilapia (Oreochromis niloticus) becoming the dominant cultured species. However, limited national surveillance has restricted early detection of parasitic infestations that could indicate breaches in farm-level biosecurity. This study aimed to evaluate low-cost parasitological screening as a practical indicator of biosecurity performance by assessing parasite prevalence, diversity, and predictors across inland farms during two seasons.  A sample of 30 fish from each farm was examined from 25 inland aquaculture sites sampled in summer and winter (2022–2023). External and internal parasitic infestations were evaluated through wet smears of the skin, fins, and gills, and by compression techniques for digeneans. Water quality parameters, serum cortisol levels, and farm management data were recorded. Parasites were identified to the genus-level. Predictors of infestation were analyzed using binary logistic regression followed by generalized linear mixed models.  Six parasite groups were detected, with infestations recorded on all farms. Trichodina spp. showed the highest farm-level prevalence (96%) and fish-level prevalence (54.3%), followed by Cichlidogyrus spp. (92% of farms; 56.9% of fish). Dactylogyrus, Ambiphyra, Gyrodactylus, and Centrocestus were variably present, with the eastern region displaying the lowest diversity. Most infestations were mild. Significant predictors varied by parasite type: summer season increased the likelihood of Centrocestus, Dactylogyrus, Cichlidogyrus, and Ambiphyra; low pH strongly predicted monogenean and sessile ciliate infestations; dissolved oxygen and fish length also contributed to parasite-specific patterns. No clear association existed between infestation and elevated cortisol levels.  This nationwide two-season assessment shows that simple parasitological screening, especially for Trichodina and Cichlidogyrus, offers quick, affordable signs of biosecurity breaches in inland O. niloticus aquaculture. The presence of parasites indicates past or current risks of pathogen introduction, highlighting the importance of better quarantine procedures, water quality management, and following national biosecurity guidelines. Regular quarterly parasitological checks, combined with water quality testing and improved farm hygiene, can enhance early detection and reduce the spread of disease across Saudi Arabia's aquaculture industry. 

Pasteurella multocida is a primary cause of hemorrhagic septicemia (HS) in buffaloes across tropical regions, leading to rapid deaths and significant economic losses. In Vietnam, recurrent HS outbreaks have been reported, yet most studies have solely focused on bacterial isolation or species-level identification. Comprehensive molecular data, including capsular typing, lipopolysaccharide (LPS) genotyping, virulence–associated genes (VAGs), and antimicrobial resistance genes (ARGs), remain limited for buffalo-derived strains. This molecular gap hampers understanding of strain diversity, epidemiology, and vaccine development. Notably, no previous Vietnamese study has concurrently characterized capsule, LPS, VAG, and ARG profiles or reported atypical serogroups in buffaloes. Therefore, integrated molecular surveillance is crucial to detect emerging lineages and guide One Health–oriented disease management. This study aimed to provide the first comprehensive molecular characterization of P. multocida strains isolated from buffaloes in Vietnam’s Central Highlands, incorporating capsular typing, LPS genotyping, virulence gene profiling, and antimicrobial resistance detection.  Sixty-seven P. multocida isolates were recovered from lungs, bone marrow, and nasal swabs of clinically affected buffaloes (2022–2025). Species confirmation and molecular screening for capsular types (A, B, D), LPS genotypes (L1–L8), 12 VAGs, and seven ARGs were performed using polymerase chain reaction-based assays. Three representative isolates underwent 16S rRNA sequencing for phylogenetic analysis. Prevalence estimates, along with their 95% confidence intervals, were calculated, and chi-square tests were performed.  Capsular type B was the most common (62.7%), followed by type A (31.3%). Importantly, capsular type D (5.9%) was identified for the first time in Vietnamese buffaloes. LPS genotyping showed L2 as the predominant genotype (56.7%), with L6 (19.4%), L1 (16.4%), and L3 (7.5%) also present. All isolates contained eight conserved VAGs, while pfhA (58.2%) and hgbB (34.3%) showed variable presence. ToxA and nanH were not detected. Four ARGs, floR (22.3%), tetB (11.9%), blaROB1 (10.4%), and tetH (4.4%), were observed. Phylogenetic analysis clustered all isolates within the P. multocida group, with the type D isolate forming a minor diverging sub-branch.  This study establishes the first multilocus molecular profile of P. multocida in Vietnamese buffaloes, highlighting the emergence of capsular type D and the dominance of L2 genotypes. These findings support better diagnostics, vaccine antigen selection, and antimicrobial stewardship. Future work using multilocus sequence typing/whole–genome sequencing across livestock species will improve understanding of regional transmission dynamics within a One Health framework. 

General anesthesia (GA) suppresses the blink reflex and lacrimal gland activity, making animals more vulnerable to precorneal tear film (PTF) issues. Although decreases in tear volume during GA are well documented, changes in PTF quality are not well understood. This study examined both the quantity and quality of PTF, including the Schirmer Tear Test-1 (STT-1), tear osmolarity (TO), tear ferning (TF), and punctate fluorescein staining (PFS), in healthy mesocephalic Canis familiaris undergoing routine non-ophthalmic surgery under GA.  A prospective, randomized, pre–post study was conducted on 16 client-owned mesocephalic dogs (32 eyes). All subjects were clinically and ophthalmologically normal and classified as American Society of Anesthesiologists (ASA) I–II. Tear film parameters were evaluated at five perioperative time points: 30 min preoperatively (T0), 10 min post-premedication (T10), 5 min post-induction (T5), at first surgical incision (TS), and at discharge (TD). STT-1, TF, and TO were measured at each time point; PFS was performed at TD. GA consisted of methadone and dexmedetomidine premedication, propofol induction, and isoflurane maintenance. Mixed-effects regression, paired t-tests, and correlation analyses were applied, with p < 0.05 considered significant.  STT-1 values significantly decreased from baseline (21.2 ± 3.3 mm/min) to T10 (13.5 ± 5.9 mm/min; p < 0.001), T5 (6.4 ± 6.3 mm/min; p < 0.001), and TS (0.8 ± 1.6 mm/min; p < 0.001). TO decreased from 374.4 ± 29.3 mOsm/L at T0 to 354.7 ± 28.2 mOsm/L at TS (p < 0.001). TF grades increased from 0.8 ± 1.0 at T0 to 1.5 ± 1.3 at T10 and 2.3 ± 1.4 at T5 (p < 0.001), indicating deterioration of PTF structure. Moderate correlations were observed among STT-1, TF, and TO. At TD, tear parameters remained significantly altered compared with T0, and PFS identified punctate epithelial lesions in 34.4% of dogs. Age showed a moderate negative relationship with STT-1 (b = –0.41 mm/min; p = 0.038).  GA causes a significant decline in the quantity and quality of the PTF, with incomplete recovery by discharge despite the return of spontaneous blinking. These findings emphasize the need for proactive perioperative ocular surface protection and highlight TF and TO as useful early indicators of anesthesia-related ocular surface impairment in mesocephalic Canis familiaris. 

Milk yield and composition in goats are heavily influenced by feeding and management practices, especially in arid areas where forage availability varies. Native goats in northern Mexico are well adapted to tough environments, yet there is limited evidence on how different production systems impact their milk quality. This study aimed to compare milk yield, chemical makeup, and the fatty acid (FA) profile, particularly conjugated linoleic acid (CLA), of early-lactation native goats managed under grazing and housed systems.  Ten clinically healthy native goats in early-lactation were randomly assigned to two treatments: grazing (n = 5) and housed (n = 5). Housed goats received a mixed ration, while grazing goats foraged daily on native rangeland. Milk yield was recorded weekly, and 100 mL composite milk samples were analyzed for fat, protein, lactose, and FA profile using infrared spectrophotometry and gas chromatography. The experimental period included 14 days of adaptation and 42 days of data collection. Data were analyzed using a completely randomized design with repeated-measures in PROC MIXED (SAS v9.4), and Tukey’s test was applied for comparisons (p < 0.05).  Housed goats showed significantly higher (p < 0.05) daily milk yield (1.41 vs. 0.81 kg per day) and fat-corrected milk (1.22 vs. 0.83 kg per day). Protein and lactose concentrations were also greater in housed goats. In contrast, grazing goats produced milk with higher fat concentration (+42.3%; p = 0.0238) and a more favorable FA profile, including increased unsaturated FAs such as oleic acid (C18:1 c9), α-linolenic acid (C18:3 n-3), and cis-9, trans-11 CLA (p = 0.0009). Grazing also decreased medium-chain saturated FAs while increasing long-chain FAs and total monounsaturated FAs.  Grazing boosts the nutritional quality of milk from native goats by increasing unsaturated FAs and significantly raising CLA levels, though it comes with a reduction in milk volume. These results underscore the importance of native goats and extensive grazing systems for producing nutrient-rich milk in arid areas, supporting both sustainability and potential markets for functional foods. 

Postpartum infertility remains a major challenge in tropical dairy systems, particularly in Zebu crossbred cows that experience delayed ovarian cyclicity and increased metabolic and environmental stress. Improving early conception is essential to reducing calving intervals and enhancing productivity. This study evaluated the effectiveness of an Augmented-OvSynch protocol incorporating a post-insemination gonadotropin-releasing hormone (GnRH) injection in improving reproductive performance and assessing associated endocrine and oxidative stress dynamics in postpartum Zebu crossbred cows under field conditions.  A total of 219 clinically healthy pluriparous Zebu crossbred cows (Holstein-Friesian × Sahiwal), 55–90 days postpartum, were randomly allocated to either a control group receiving the standard OvSynch protocol (n = 82) or a treatment group receiving the Augmented-OvSynch protocol (n = 137), which included an additional GnRH dose on day 5 post-artificial insemination (AI). Estrus response, follicular dynamics, pregnancy rate, and pregnancy loss were monitored through behavioral signs and ultrasonography. Blood samples collected on days 0, 5, 10, and 21 were analyzed for estrogen, progesterone, malondialdehyde (MDA), superoxide dismutase (SOD), and total antioxidant capacity (TAC). Statistical significance was set at p < 0.05.  The Augmented-OvSynch protocol significantly improved estrus expression (83.94%) compared with the control group (37.80%). Pregnancy rate per AI was markedly higher in treated cows (68.61%) than in controls (29.27%). Progesterone concentrations increased significantly from day 5 to day 10 in treated cows, indicating enhanced luteal support and probable accessory corpus luteum formation. Estrogen levels declined more sharply in pregnant cows. Oxidative stress markers were more strongly associated with pregnancy status than protocol type: pregnant cows consistently showed lower MDA and higher TAC levels, whereas SOD exhibited moderate variation without strong treatment effects.  The Augmented-OvSynch protocol substantially improved reproductive outcomes in postpartum Zebu crossbred cows, primarily by enhancing luteal activity and improving synchronization responses. Although oxidative stress markers were influenced more by pregnancy outcome than treatment, their integration provided valuable insight into physiological determinants of fertility. Owing to its simplicity, low-cost, and field suitability, the protocol offers a practical and scalable reproductive management strategy for tropical smallholder dairy systems. 

Corn is the main cereal used in broiler nutrition because of its high energy content and carotenoid richness, while wheat and barley offer alternative nutrient profiles that may increase production flexibility. However, their relative impacts on broiler performance, carcass traits, meat physicochemical properties, and consumer sensory perception under standardized enzyme-supplemented conditions remain unclear. This study examined the effects of partially replacing corn with wheat or barley on growth performance, carcass yield, meat quality, and consumer sensory evaluation in broiler chickens.  A total of 525 male ROSS 308 broilers were randomly assigned to three dietary treatments: corn (C), corn–wheat (CW), and corn–barley (CB), with five replicates of 35 birds each. Diets were isocaloric and isonitrogenous and supplemented with a xylanase–β-glucanase complex. Birds were raised for 35 days under controlled environmental conditions. Growth performance, carcass traits, meat color (L*, a*, b*), pH, water-holding capacity, texture profile, and consumer sensory attributes were evaluated using standardized protocols. Data were analyzed using General Linear Model procedures with significance set at p < 0.05.  Broilers fed with corn showed numerically higher body weight gain (+4.8% compared to CW; +4.4% compared to CB) and a tendency toward improved feed conversion ratio (1.52 vs. 1.56–1.58; p = 0.10). Including barley significantly increased abdominal fat (+36% vs. corn; p = 0.04), while wheat resulted in the lowest fat deposition. Meat yellowness (b*) was highest in the corn group at both 45 min and 24 h postmortem (p < 0.05, p < 0.01), reflecting the higher carotenoid content of corn. No significant differences were found among treatments for pH, drip loss, cooking and thawing loss, texture parameters, or sensory scores (p > 0.05). All sensory attributes scored above 4.3 on the 7-point scale.  Moderate inclusion of wheat (12%–20%) or barley (8%–15%) in enzyme-supplemented diets did not affect growth performance, carcass yield, meat physicochemical traits, or consumer sensory acceptance. Wheat might be used strategically to reduce abdominal fat, while corn remains preferred when enhanced yellowness is desired. These findings support the practical use of wheat and barley as viable alternatives to corn in commercial broiler feeding programs. 

Bovine trichomonosis, caused by Tritrichomonas foetus, is a significant reproductive disease that impacts cattle productivity and breeding efficiency. In South Africa, routine diagnostic methods often depend on culture and microscopy, which may not accurately distinguish T. foetus from nonpathogenic trichomonads. This study aimed to determine the prevalence of T. foetus in bulls from the Dr. Segomotsi Ruth Mompati (DSRM) District, North-West Province, South Africa, using advanced molecular diagnostics, including real-time polymerase chain reaction (RT-PCR), conventional PCR, DNA sequencing, and phylogenetic analysis.  A total of 239 sheath wash samples were collected between June 2018 and October 2020. Of these, 51 culture-positive trichomonad isolates were selected for molecular analysis. Microscopy and modified Giemsa staining were used to characterize protozoal morphology. DNA was extracted and subjected to RT-PCR with 5’ TaqMan™ probes, as well as conventional PCR targeting the 5.8S rRNA/Internal Transcribed Spacer (ITS) regions. PCR amplicons were sequenced, and phylogenetic trees were constructed using MEGA (maximum-likelihood, 1,000 bootstrap replicates). Statistical comparisons between diagnostic methods were performed using Chi-square and Cochran’s Q test.  RT-PCR detected T. foetus in 80.4% (41/51) of the culture-positive samples, with most isolates showing low Ct values, indicating strong positivity. Conventional PCR successfully amplified 12 isolates (300–340 bp), all of which were confirmed as T. foetus by sequencing. Phylogenetic analysis showed that the isolates clustered with the Southern African genotype, exhibiting 77%–87% similarity to Namibian strains and were closely related to Australian and Turkish isolates. No significant correlation was found between geographic location and PCR positivity. RT-PCR demonstrated significantly higher sensitivity than conventional PCR (p < 0.05).  This study confirms a high prevalence of T. foetus in bulls in the DSRM district and demonstrates the superior accuracy of molecular diagnostics compared with culture and microscopy. The identification of genotypes closely related to Southern African strains highlights potential transboundary spread. Incorporating PCR-based screening into routine surveillance is essential for accurate diagnosis, minimizing unnecessary culling, and enhancing reproductive herd health. Further longitudinal studies are recommended to assess disease dynamics and inform regional control programs.