Gamma-oryzanol, a bioactive compound derived from rice bran, is recognized for its antioxidant and metabolic regulatory properties. This study evaluated the effects of dietary gamma-oryzanol supplementation on laying performance, egg quality, blood health, and follicular development in Japanese quails (Coturnix japonica). A total of 216 12-week-old laying quails were randomly assigned to six groups receiving gamma-oryzanol at 0 (control), 0.1, 0.2, 0.3, 0.4, and 0.5 g/kg of diet for 7 weeks. Feed intake, egg production, feed conver­sion ratio, egg quality traits, hematological and biochemical parameters, and ovarian follicular characteristics were mea­sured. Statistical analysis was conducted using one-way analysis of variance and Duncan’s post hoc test. Gamma-oryzanol at 0.2 g/kg significantly increased egg production (98.69%) compared to the control (86.80%). Supplementation at 0.3–0.5 g/kg improved eggshell thickness and yolk index (p < 0.05). Red blood cell counts were signifi­cantly higher at 0.4 g/kg, indicating enhanced erythropoiesis. Other hematological and biochemical parameters remained unchanged. A significant increase in small white follicle count and F3 follicle diameter was observed at 0.5 g/kg, indicating improved follicular development. Gamma-oryzanol supplementation at 0.2–0.5 g/kg enhanced laying performance, egg quality, and ovarian fol­licle development without adverse hematological or biochemical effects. The compound shows potential as a safe and functional feed additive to support productivity and reproductive efficiency in commercial quail production.

Bovine tuberculosis (bTB), caused by Mycobacterium bovis, remains a significant zoonotic and eco­nomic threat globally. Despite the long-standing use of the Bacillus Calmette-Guérin (BCG) vaccine, its inconsistent efficacy and interference with surveillance tests underscore the need for alternative approaches. This study evaluated the safety, immunogenicity, and protective efficacy of a novel influenza vector-based vaccine expressing M. bovis antigens ESAT-6 and TB10.4, formulated with or without an adjuvant. Recombinant influenza A viruses expressing ESAT-6 and TB10.4 were constructed using reverse genetics and incorporated into vaccine formulations. Guinea pigs and calves were immunized with adjuvanted and non-adjuvanted formulations, followed by challenge with a virulent M. bovis strain. Safety was assessed through clinical observation and histopathology. Immune responses were monitored using interferon-gamma (IFNγ) enzyme-linked immu­nosorbent assay, and protection was evaluated through organ damage indices, bacterial load, and survival rates over a 12-month period. Both formulations were safe and well-tolerated in guinea pigs and calves, with no adverse clinical signs. The non-adjuvanted vaccine induced the highest and most sustained IFNγ response, peaking between 2 and 5 months post-vac­cination. In guinea pigs, the protection index reached +0.60 lg in the non-adjuvanted group versus +0.2 lg in the adjuvanted group. In calves, lung bacterial load was reduced to 1.83–1.93 lg colony-forming unit (CFU) in vaccinated animals compared with 5.8 lg CFU in unvaccinated controls. Histopathological examination confirmed minimal tissue damage in the vaccinated groups. Both vaccine formulations demonstrated protective efficacy equivalent to or better than BCG, with the non-adju­vanted version showing superior performance. This novel influenza vector-based vaccine expressing ESAT-6 and TB10.4 antigens elicits strong, long-lasting cellular immunity and provides significant protection against M. bovis infection in guinea pigs and calves. The adjuvant-free formulation demonstrated higher immunogenicity, simplified production, and minimal adverse reactions, positioning it as a promising alternative to BCG for bTB control in livestock.

Pregnancy induces significant anatomical and physiological changes, many of which are regulated by the autonomic nervous system (ANS). Heart rate variability (HRV) is a well-established non-invasive tool for assessing ANS activity. While changes in heart rate (HR) and HRV during the third-trimester of equine pregnancy are documented, there is limited understanding of cardiac autonomic adaptations during the early stages of gestation. This study aimed to compare HR and time-domain HRV parameters between healthy non-pregnant mares and those in the first and second trimesters of pregnancy. A total of 45 Thai native crossbred mares were enrolled and divided into three groups: Non-pregnant (n = 5), first-trimester pregnant (0–114 days; n = 18), and second-trimester pregnant (115–226 days; n = 22). All mares were clinically healthy and free from cardiac abnormalities. Electrocardiographic data were collected using a Holter electrocardiogram system over a 15 min period at rest, and HRV was analyzed using time-domain measures: Standard deviation of all NN intervals (SDNN), SDNN index, root mean square of successive differences, standard deviation of 5-min mean NN intervals, percentage of successive NN intervals >50 ms, and vasovagal tonus index (VVTI). Data were analyzed using Kruskal–Wallis and Mann–Whitney U-tests. HR was significantly higher in first-trimester pregnant mares compared to non-pregnant mares (p < 0.05), and even higher in the second-trimester compared to the first (p < 0.05). However, there were no significant differences among the groups in any of the HRV parameters or VVTI. The findings indicate that cardiovascular adaptation during early pregnancy in mares is characterized by a progressive increase in HR, likely reflecting increased cardiac output to support fetal development. However, the lack of significant changes in time-domain HRV parameters and VVTI suggests that ANS balance is maintained during the first and second trimesters. These results provide valuable reference values for equine reproductive monitoring and contribute to a better understanding of physiological changes in early gestation.

Coriandrum sativum L. (coriander) has long been valued for its culinary and medicinal uses. C. sativum essential oil (CsEO), particularly linalool-rich chemotypes, exhibits diverse biological activities; however, integrated evalu­ations encompassing neurological, inflammatory, and molecular targets remain limited. This study aimed to chemically characterize Peruvian CsEO and assess its anticonvulsant, analgesic, anti-inflammatory, and antioxidant effects, alongside those of pure linalool, while elucidating potential mechanisms through cytokine modulation and molecular docking of cyclooxygenase (COX) enzymes. CsEO was extracted from Peruvian coriander seeds through steam distillation and analyzed using gas chromatography–mass spectrometry (GC-MS). Antioxidant activity was quantified using the 2,2′-azino-bis(3-ethylbenz­thiazoline-6-sulfonic acid) assay. Anticonvulsant effects were tested in BALB/c mice using the pentylenetetrazole-induced seizure model, analgesic activity through the acetic acid-induced writhing test, and anti-inflammatory effects in Holtzman rats using the carrageenan-induced paw edema model. Serum interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels were measured by enzyme-linked immunosorbent assay. Molecular docking evaluated linalool’s binding affinity to COX-1 and COX-2 relative to standard non-steroidal anti-inflammatory drugs. GC-MS identified linalool as the major constituent (59.80%), alongside α-pinene (8.65%), camphor (8.48%), and γ-terpinene (7.09%). CsEO demonstrated potent antioxidant activity (half-maximal inhibitory concentration [IC50] = 32.04 μg/mL), exceeding that of linalool alone (IC50 = 152.29 μg/mL). Significant anticonvulsant effects occurred at 200 mg/kg for both CsEO and linalool, increasing seizure latency by up to 87.20% and reducing seizure frequency by ~43%. In analgesic assays, linalool (200 mg/kg) achieved a 93.80% writhing reduction, comparable to tramadol, while CsEO showed strong but slightly lower efficacy. CsEO (200 mg/kg) inhibited carrageenan-induced edema by 51.35% at 4 h, reduced IL-1β by 49.8%, and IL-6 by 26.5%, effects comparable to ibuprofen. Docking revealed moderate linalool affinity for COX-1 (−5.70 kcal/mol) and COX-2 (−6.10 kcal/mol), sharing key hydrophobic interactions with reference drugs. Peruvian CsEO, characterized by a distinctive linalool-rich chemotype, exhibits significant multi-target pharma­cological activities, with synergistic contributions from minor constituents enhancing antioxidant and anti-inflammatory effects. Its integrated efficacy profile and favorable safety indicators highlight CsEO as a promising phytotherapeutic candi­date for managing seizures, pain, and inflammation. Further studies should explore chronic models, pharmacokinetics, and formulation strategies to optimize clinical applicability.

Stallion sexual behavior during semen collection can be influenced by multiple factors, yet the role of sexual arousal intensity remains underexplored. Understanding how arousal modulates behavioral and physiological reproductive traits is essential for improving artificial insemination (AI) efficiency and semen quality. This study aimed to evaluate the effects of sexual arousal intensity, age, origin, and semen collection method on stallion sexual behavior and the quality of fresh and frozen semen. Thirteen Arabian stallions (7 Tunisian, 6 foreign parentage) aged 6–20 years (total ejaculates = 49) were assessed during semen collection using either a dummy or an estrous mare. Sexual arousal intensity was scored on a four-point scale (− to +++). Behavioral responses, mounting and erection parameters, and semen traits were recorded. Fresh semen was evaluated for volume, motility, concentration, and morphology; frozen semen was assessed for motility, viability, membrane integrity, and abnormalities. Data were analyzed using multifactorial analysis of variance with signifi­cance at p < 0.05. High arousal (+++) increased vocalizations, anogenital sniffing, and Flehmen responses (p < 0.05) but prolonged preparation time (p = 0.05). Low arousal (−/+) prolonged full erection duration (p < 0.01) and improved sperm motility in fresh and frozen semen (p < 0.01). Stallions of foreign origin exhibited higher fresh semen motility (p < 0.01) but required longer preparation and collection times (p < 0.05). The estrous mare method improved fresh semen motility and concen­tration (p < 0.05) but did not affect frozen semen traits. Younger stallions produced larger semen volumes, but had higher abnormal sperm counts in fresh samples, whereas older stallions showed more abnormalities post-freezing (p < 0.01). Sexual arousal intensity significantly modulates both behavioral and semen quality parameters in stallions. Lower arousal is associated with prolonged erection and superior sperm motility, suggesting dissociation between behav­ioral excitement and physiological semen traits. Collection from an estrous mare can enhance fresh semen motility, though the dummy remains safer for handlers. These findings highlight the importance of tailoring semen collection protocols to individual stallion profiles to optimize AI outcomes.

Rabbits are increasingly valued in tropical livestock systems for their efficient feed conversion, high-quality meat, and adaptability to small-to-medium scale farming. Genetic selection using molecular markers, such as single-nucleotide polymorphisms (SNPs) in candidate genes, offers a powerful tool to enhance carcass yield and meat quality. The melanocortin-4 receptor (MC4R) gene, a regulator of energy balance and feed intake, has been associated with growth and carcass traits in various livestock but remains underexplored in tropical rabbit production systems. This study aimed to investigate the association between MC4R SNPs and economically important carcass traits in three commercial rabbit breeds, New Zealand White (NZW), Hyla, and Hycole, raised under tropical conditions in Indonesia. Thirty-five male rabbits (10 NZW, 11 Hyla, 14 Hycole; aged 3–4 months) were selected from 621 bucks. DNA was extracted from blood samples, and a 127-base pair MC4R fragment was amplified by polymerase chain reaction and sequenced for SNP detection. Carcass traits measured included hot carcass weight, cold carcass weight, ref­erence weight, carcass percentage, meat-to-bone ratio, and five commercial cut points (CP1–CP5). Association analysis between MC4R genotypes (AA, AG, GG) and carcass traits was performed using a general linear model, considering breed and genotype × breed interactions. An SNP at position 519 (G>A) produced three genotypes. GG-genotype rabbits exhibited the highest carcass per­centage (57.0%) and superior CP2 weight (0.12 ± 0.03 kg) and percentage (10.67 ± 1.53%), with significant genotype effects (p < 0.05) for CP2 weight and CP1 percentage. Breed effects were significant for carcass percentage and CP1 percentage (p < 0.05), while genotype × breed interactions significantly influenced CP2 traits (p < 0.01). The G allele showed a positive association with forequarter meat yield, suggesting its utility in marker-assisted selection (MAS). MC4R polymorphism at position 519 is significantly associated with key carcass traits in commercial rabbits under tropical conditions. The GG genotype is linked to improved carcass yield, particularly in CP2 traits, although effects vary by breed. Incorporating this SNP into MAS programs can enhance meat production efficiency and carcass quality in tropical rabbit breeding.

Leptospirosis is a globally neglected zoonosis caused by pathogenic spirochetes of the genus Leptospira. It affects a wide range of animals and poses serious public and veterinary health risks. Backyard cattle systems, common across Latin America, are particularly vulnerable due to poor biosecurity and close animal–human–wildlife inter­faces. This study assessed the seroprevalence, spatial distribution, and risk factors associated with Leptospira infection in backyard cattle herds of central Chiapas, Mexico. A cross-sectional study was conducted from January to September 2022 across five municipali­ties. A total of 590 serum samples were collected and analyzed using the microscopic agglutination test with six serovars. The geographical coordinates of production units (PUs) were recorded using a Global Positioning System (GPS), and spatial analysis was conducted with ArcGIS 10.5. Risk factor associations were evaluated through Chi-square tests and multivariate logistic regression using RStudio and Statistical Package for the Social Sciences software. The overall seroprevalence of Leptospira was 27.72% (95% confidence interval [CI]: 23.97%–31.75%), with the Portland Vere serovar being predominant (22.89%). Cintalapa exhibited the highest municipal seroprevalence (61.75%; odds ratio [OR] = 6.2). Logistic regression identified significant risk factors for seropositivity, including artificial insemination (OR = 2.43), use of Jagüey reservoirs (OR = 0.47), and cattle aged 3 years (OR = 0.54) or 8 years (OR = 1.98). The American Swiss × Holstein crossbreed was significantly associated with increased seroprevalence (OR = 3.15). The presence of dogs within PUs was significantly associated with Portland Vere seropositivity (OR = 3.82), highlighting a possible role in disease transmission. This study highlights a high burden of Leptospira interrogans serovar Canicola Portland Vere in backyard cattle of central Chiapas. Key risk factors include specific breeding methods, water sources, age, and breed. The integra­tion of serological surveillance, spatial mapping, and statistical modeling proved effective in identifying epidemiological hotspots and informing future One Health-based prevention strategies. The findings emphasize the need for enhanced disease surveillance, targeted control programs, and public health interventions tailored to small-scale cattle systems in tropical regions.

Foot-and-mouth disease (FMD) is a highly contagious transboundary animal disease affecting clo­ven-hoofed livestock, with significant economic and trade implications. Armenia lies within the West Eurasia and Middle East epidemiological pool, where serotypes O, A, Asia-1, and occasionally SAT-2 circulate. Despite decades of control efforts, the historical epidemiology of FMD in Armenia has not been comprehensively documented. This study aimed to conduct a 65-year retrospective analysis of FMD in Armenia to characterize serotype distribution, outbreak patterns, vaccination strat­egies, and diagnostic advancements, and to identify priorities for progression in the progressive control pathway (PCP-FMD). Data from 1958 to 2023 were compiled from the Union of Soviet Socialist Republics (USSR) agri­cultural archives, national veterinary records, World Organization for Animal Health/World Reference Laboratory for FMD reports, and peer-reviewed literature. Serotype identification, outbreak frequency, and species involvement were analyzed using descriptive statistics, heatmaps, and geographic information system (GIS) mapping. Diagnostic evolution from com­plement fixation testing to enzyme-linked immunosorbent assay, reverse transcription-polymerase chain reaction, and viral protein 1 (VP1) sequencing was documented. Vaccination protocols were traced from early monovalent campaigns to cur­rent polyvalent strategies. Between 1958 and 2023, Armenia recorded over 1 million FMD cases, with peaks in 1966 (591,820 cases) and 1973 (471,263 cases). Serotypes O, A, Asia-1, and SAT-1 were detected, with serotype O predominating. Outbreaks declined significantly after the 1980s, coinciding with mass vaccination, improved diagnostics, and targeted biosecurity measures. Notable milestones included integration of the A/Armenia/98 strain into vaccines (1999) and adoption of polyvalent vac­cines containing the A/ASIA/G-VII lineage (2016). No outbreaks have been reported since 2016. Armenia’s sustained control of FMD reflects adaptive vaccination strategies, early serotype detection, and regional cooperation. Progression from PCP-FMD Stage 2 to Stage 3 will require enhanced vaccination coverage, expanded surveillance, and strengthened veterinary infrastructure. Historical lessons from Armenia’s control strategies may inform FMD management in similar transboundary risk zones.

Canine babesiosis, primarily caused by Babesia canis vogeli in Thailand, is a significant tick-borne disease of veterinary concern. Molecular diagnostics targeting the 18S rRNA gene have enhanced detection sensitivity and specificity compared to conventional methods. This study aimed to identify and characterize B. canis vogeli in naturally infected dogs in Khon Kaen, Thailand, to compare the diagnostic performance of two primer sets (Bab7/Bab9 and Babf/Babc), and to perform phylogenetic analysis of the isolates. A total of 159 ethylenediaminetetraacetic acid blood samples from client-owned dogs presented to the Veterinary Teaching Hospital, Khon Kaen University, between July and October 2024, were examined. Samples under­went Giemsa-stained blood smear microscopy and PCR amplification of the 18S rRNA gene using both primer sets. Positive amplicons were sequenced and analyzed phylogenetically using the Maximum Likelihood method. Limit of detection (LOD), sensitivity, specificity, and positive predictive value (PPV) were calculated for each primer set using sequence-confirmed results as the reference. Microscopy detected B. canis in 19/159 (11.9%) of samples, while PCR increased detection to 23/159 (14.47%). Babf/Babc detected all positive cases (100% sensitivity), while Bab7/Bab9 detected 95.65% of positives. Both primer sets achieved 100% specificity and PPV, with an equal LOD of 105 DNA copies. Bab7/Bab9 also amplified Hepatozoon canis at a distinct amplicon size (503 base pair). Sequence analysis confirmed all Babesia-positive samples as B. canis vogeli, showing 96.34%–100% identity with global isolates. Phylogenetic analysis grouped the sequences with B. canis vogeli from multiple geographic regions, revealing minimal intraspecific variation. B. canis vogeli was the only subspecies identified in naturally infected dogs in Khon Kaen during the study period. Babf/Babc demonstrated superior diagnostic sensitivity for B. canis vogeli, whereas Bab7/Bab9 offered broader detection, including H. canis. Phylogenetic analysis revealed close genetic relationships with isolates worldwide. These find­ings support the use of Babf/Babc for specific diagnosis and Bab7/Bab9 for broader screening in endemic regions.

Insect-derived proteins are gaining attention as sustainable pet food ingredients, but the use of cricket protein hydrolysate (CPH) in canine diets remains underexplored. This study evaluated the effects of CPH on diet palatability, physiological responses, and antioxidant potential for shelf-life extension in commercial dog food. Thirty-two healthy adult dogs were assigned to four diets containing 0%, 2%, 4%, or 6% CPH for a 30-day feeding trial. Palatability was assessed through a two-bowl preference test, while biochemical, hematological, and fecal parameters were measured pre- and post-trial. Antioxidant efficacy was evaluated by monitoring acid value (AV) and peroxide value (PV) during accelerated storage (55°C for 46 days, simulating 12 months). Nutritional adequacy was con­firmed through proximate and amino acid analysis. The 2% CPH diet significantly improved palatability, with a 57% increase in intake compared to control (p < 0.05), whereas higher inclusions (4% and 6%) reduced acceptance due to bitterness from hydrophobic peptides. All health param­eters remained within reference ranges, though the 6% CPH diet lowered serum glucose (87.0 vs. 112.0 mg/dL; p < 0.001) and increased blood urea nitrogen (11.0 mg/dL; p = 0.0023). Antioxidant activity increased with CPH level, with 6% CPH reducing PV by 33% after 46 days (p < 0.05). CPH lacked certain essential amino acids, notably tryptophan, requiring com­plementary protein supplementation. CPH is a multifunctional ingredient that can enhance palatability and oxidative stability in dog diets at moderate inclusion (2%). High inclusion levels improve antioxidant capacity but may impair sensory acceptance and alter metabolic markers. Long-term safety, allergenicity, and flavor-masking strategies warrant further study.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric pathogen causing severe diar­rhea and high mortality in neonatal piglets. Maternal lactogenic immunity, conveyed through colostrum and milk, is essen­tial for protection; however, sow-derived antibodies may be insufficient in certain production systems. This study aimed to produce PEDV-specific hyperimmune colostrum and milk from goats and evaluate the safety and immunogenicity of live-attenuated and inactivated PEDV vaccines. Preliminary safety trials were performed in male goats (n = 6) to monitor clinical signs and adverse reactions after intramuscular vaccination. Ten pregnant Saanen goats were randomly assigned to two groups (n = 5 each) and immunized twice, 8 and 4 weeks before parturition, with either live-attenuated (1 × 105 50% tissue culture infectious dose [TCID50]/mL) or inactivated (1 × 106 TCID50/mL) PEDV vaccine. Serum was collected on days 0 and 28 post-vaccination, and colostrum/milk samples were obtained on days 0, 2, 7, and 14 postpartum for virus neutralization (VN) assays. Fecal samples were analyzed using quantitative reverse transcription polymerase chain reaction to detect viral shedding. Both vaccines were well tolerated, with only transient fever observed in three goats. No severe adverse reac­tions occurred in pregnant goats. The live-attenuated vaccine elicited higher VN titers in serum (peak 1:32) and colostrum (peak 1:2048) compared with the inactivated vaccine (serum peak 1:16; colostrum peak 1:512). Day 0 colostrum titers were significantly greater in the live vaccine group (p = 0.028). Although titers remained higher in this group on days 2, 7, and 14, differences were not statistically significant. Viral RNA shedding was absent in the inactivated group and transient in the live group, persisting up to 7 days in one goat. Vaccination of pregnant goats with live-attenuated or inactivated PEDV vaccines is safe and induces PEDV-specific antibodies in colostrum and milk. The live-attenuated vaccine generated the highest titers, indicating potential for producing goat-derived hyperimmune colostrum as an alternative passive immunization strategy for neonatal piglets. Field trials are warranted to confirm protective efficacy and explore integration into PEDV control programs.

Enhancing poultry reproductive performance is essential for improving productivity and addressing protein shortages in developing regions. Yoruba ecotype chickens (YECs) are resilient but limited in commercial potential due to small body size and low egg yield. This study assessed the fertilizing potential of YEC × Sussex (SS) crossbred cocks by integrating conventional reproductive morphometrics with seminal plasma metabolite profiling to identify potential biomarkers of sperm quality. Thirty 24-week-old YEC × SS cocks (2842.86 ± 137.33 g) were evaluated over 28 days. Semen was collected through abdominal massage, and semen volume, sperm concentration, and testosterone levels (enzyme-linked immunosorbent assay) were recorded. Testicular and epididymal morphometrics, densities, and sperm reserves were mea­sured. Seminal plasma metabolites were analyzed using high-performance liquid chromatography-mass spectrometry. Pearson correlations and linear regression models were applied to predict testosterone concentration from semen and morphometric traits. The left testis was heavier (8.00 g) and larger in volume (12.77 mL) than the right (6.75 g; 1.35 mL). Spermatozoa reserves averaged 0.20 × 109, with a strong positive correlation with testis volume (r = 0.998, p < 0.01) and a moderate neg­ative correlation with daily sperm production (r = –0.585, p < 0.01). Testosterone concentration prediction from live weight, semen volume, and sperm concentration achieved high accuracy (R2 = 0.829). Thirteen seminal plasma metabolites were identified, including ascorbic acid, quercetin, epicatechin, citric acid, and procyanidin B2 – compounds linked to antioxidant defense, energy metabolism, and sperm viability. YEC × SS crossbred cocks exhibit favorable reproductive morphometrics, strong correlations between testis vol­ume and sperm reserves, and a metabolite profile enriched in fertility-enhancing antioxidants. Predictive models using basic semen traits can reliably estimate testosterone levels, while identified metabolites have potential as biochemical markers for breeding selection. Integrating morphometric and metabolomic profiling can refine breeding strategies, improve artifi­cial insemination outcomes, and enhance the genetic improvement of local poultry breeds.