Background and Aim:Bluetongue virus (BTV), an arbovirus of major economic importance, affects domestic and wild ruminants globally and is primarily transmitted by Culicoides biting midges. The virus is endemic in many regions, yet limited data are available for Saudi Arabia. This study aimed to determine the seroprevalence of BTV antibodies in cattle, goats, sheep, and camels across two ecologically distinct regions in central Saudi Arabia and to assess species- and region-specific risk profiles.Materials and Methods:A total of 1,194 serum samples were collected from apparently healthy livestock (280 cattle, 159 camels, 429 sheep, and 326 goats) in Riyadh and Al-Qassim between October 2023 and March 2024. Samples were tested for BTV antibodies using a commercial competitive enzyme-linked immunosorbent assay. Statistical analysis included Chi-square tests and odds ratios with 95% confidence intervals to compare prevalence rates between species and regions.Results:The overall BTV seroprevalence was 44.6% (533/1,194). Goats had the highest prevalence (59.8%), followed by cattle (51%), sheep (36.3%), and camels (22.6%). In Riyadh, cattle (55.7%) and goats (55%) showed the highest rates, while in Al-Qassim, goats (65.7%) were most affected. Camels consistently showed the lowest seroprevalence (18.6%–25%). Statistically significant differences in seroprevalence were observed among species and between regions (p < 0.05).Conclusion:BTV is endemic in central Saudi Arabia, with substantial species and regional variability. Goats and cattle are at higher risk, indicating a need for species-targeted surveillance and vector control. The findings support the implementation of national bluetongue monitoring strategies and lay the groundwork for future molecular and longitudinal studies.

Background and Aim: Antibiotic resistance has spurred interest in alternative feed additives for poultry. Wood vinegar (WV), a by-product of plant pyrolysis, contains bioactive compounds with antioxidant and antimicrobial properties. This study aimed to evaluate the effects of WV supplementation through drinking water on the cecal microbial population, volatile fatty acid (VFA) concentrations, antioxidant enzyme activity, and apparent ileal nutrient digestibility in broiler chickens. Materials and Methods: A total of 432 1-day-old male Cobb 500 broiler chicks were randomly assigned to six groups (n = 72 per group; 6 replicates of 12 birds each). Treatments included a negative control (T1), a positive control with 0.02% oxytetracycline (T2), and WV-supplemented groups at dilution ratios of 1:100 (T3), 1:200 (T4), 1:500 (T5), and 1:1000 (T6) in drinking water. The experiment lasted 35 days. Plasma antioxidant enzymes (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx], total antioxidant capacity [T-AOC]), VFA profiles, ileal digestibility (crude protein [CP], ash, ether extract [EE]), and cecal microbial populations were assessed. Statistical analysis was performed using the General Linear Model and Duncan’s multiple range tests (p &lt; 0.05). Results: WV supplementation enhanced antioxidant status, with significant increases in GPx (T5 and T6) and T-AOC (T6), while CAT and SOD remained unaffected. T5 significantly elevated acetic, butyric, and total VFA levels. WV-treated birds (T3–T6) showed reduced Salmonella, Escherichia coli, and Enterobacteria counts and increased bifidobacteria and total bacteria compared with controls. T4 showed the highest digestibility of CP, while T5 significantly improved ash and EE digestibility. Conclusion: WV supplementation, particularly at a 1:200 dilution (T4), effectively improved gut microbial balance, enhanced antioxidant enzyme activity, and promoted nutrient digestibility. These results support WV as a viable natural alternative to antibiotic growth promoters in broiler production. Keywords: antibiotic alternative, antioxidant enzymes, broiler chickens, cecal microbiota, nutrient digestibility, volatile fatty acids, wood vinegar.

Background and Aim:Zoonotic malaria remains a significant public health concern in Southeast Asia. The potential role of cattle as reservoirs for Plasmodium spp. in Indonesia has not been fully elucidated, despite increasing recognition of animal reservoirs in malaria transmission dynamics. This study aimed to investigate the prevalence of Plasmodium spp. in humans and cattle in a malaria-endemic region of Indonesia to explore the potential for zoonotic transmission and inform integrated control strategies aligned with Sustainable Development Goal 3.3.Materials and Methods:A cross-sectional study was conducted between March to July 2024 involving 41 human participants and 43 cattle. Blood samples were collected and analyzed using endpoint polymerase chain reaction techniques targeting Plasmodium genus-specific DNA sequences. The infection prevalence in both populations was determined, and the results were interpreted to assess the risk of zoonotic malaria transmission.Results:All human blood samples tested negative for Plasmodium spp., corresponding to a 0% infection rate (95% confidence interval [CI]: 0.0%–8.5%). In contrast, one cattle sample tested positive, resulting in a 2.33% infection rate among cattle (95% CI: 0.06%–12.0%). The positive detection in cattle was confirmed by a distinct 240 base pairs band through agarose gel electrophoresis. The absence of infections in humans suggests the effectiveness of current public health measures, while the presence of Plasmodium DNA in cattle underscores the potential role of cattle as parasite reservoirs.Conclusion:The findings highlight the importance of integrating animal health surveillance into malaria elimination programs under the One Health framework. Although no zoonotic transmission to humans was observed, the detection of Plasmodium spp. in cattle warrants continuous surveillance, improved livestock management practices, and targeted vector control measures. Further studies with species-specific molecular diagnostics and broader geographic coverage are recommended to clarify the zoonotic potential and transmission dynamics involving cattle.

Background and Aim:The leptin (LEP) gene plays a pivotal role in regulating growth, metabolism, and fat deposition in cattle. Genetic polymorphisms in this gene can influence phenotypic traits and may serve as molecular markers for selection in breeding programs. However, comprehensive characterization of LEP gene variants in local Indonesian breeds, such as Madura cattle, remains limited. This study aimed to identify novel single-nucleotide polymorphisms (SNPs) within exon 2 of the bovine LEP gene and assess their association with growth traits in Madura cattle.Materials and Methods:Forty-five Madura cows (aged 2–4 years) were phenotypically evaluated for body weight (BW), wither height (WH), body length (BL), chest girth (CG), hip height (HH), head length (HL), and head width (HW). Genomic DNA was extracted and subjected to polymerase chain reaction amplification followed by Sanger sequencing. Detected SNPs were analyzed for genotype and allele frequencies, Hardy–Weinberg equilibrium (HWE), and their associations with growth traits using a general linear model.Results:Three SNPs were identified in exon 2 of the LEP gene: c.126T>C (synonymous), c.148T>C (missense), and a novel missense SNP c.149G>A, resulting in a cysteine to tyrosine substitution at amino acid position 50. The c.149G>A SNP showed polymorphism with three genotypes (GG, GA, AA), and the heterozygous GA genotype had the highest frequency (64.4%). This SNP deviated from HWE (p < 0.05), indicating potential selection pressure or population structure effects. While no statistically significant associations were found between genotypes and growth traits (p > 0.05), the AA genotype showed the highest mean values across most body measurements.Conclusion:Although the novel c.149G>A SNP was not significantly associated with growth traits, its polymorphic nature and descriptive trait patterns suggest it may have biological relevance. Larger-scale studies are recommended to validate its utility as a genetic marker for growth and development in Madura cattle.

Salmonella Abortusequi is a significant etiological agent of equine abortions, yet limited genomic data exist, particularly in Central Asia. This study aimed to perform the first genome-wide characterization and phylogenetic analysis of three S. Abortusequi strains isolated from equine abortions in different regions of Kazakhstan. Whole-genome sequencing was conducted on three isolates using the Illumina MiSeq platform. Genomic assemblies were annotated using SPAdes and Prokka, while phenotypic traits were predicted through BioNumerics. Antimicrobial resistance genes, virulence factors, and prophage elements were identified using established databases. Phylogenetic relationships were examined through whole-genome single-nucleotide polymorphism (wgSNP) analysis against a global panel of S. Abortusequi and related serovars. All isolates displayed high genomic similarity and were classified as Salmonella enterica subsp. enterica serovar Abortusequi with an antigenic profile of 4:a:e,n,x. Twelve Salmonella pathogenicity islands and three prophages were identified, with ST64B present in all isolates. The ac(6')-Iaa gene, which confers resistance to aminoglycosides, was detected in all strains. Each genome encoded 101-109 virulence factors, with 94 conserved across isolates. wgSNP analysis confirmed close phylogenetic clustering of the Kazakh strains, with regional variation between northern and southern isolates. Prophage-associated virulence elements, particularly virulence factor protein (SseK), were also documented. This study reveals the genetic uniformity and virulence potential of S. Abortusequi strains circulating in Kazakhstan. The presence of conserved resistance and virulence genes, including prophage-encoded elements, underscores the pathogenic risk posed by these isolates. These findings contribute valuable genomic data for surveillance, diagnosis, and control of salmonellosis in equine populations. Despite the limited sample size, the study establishes a foundation for future genomic epidemiological studies and targeted disease mitigation strategies.

Industrial-scale probiotic production requires economically viable media formulations that do not compromise strain functionality. This study aimed to develop a cost-effective medium for cultivating Lactiplantibacillus plantarum 22F (L22F), a probiotic candidate isolated from swine feces, while evaluating its industrial viability and functional metabolic profile. Carbon (glucose, sucrose, and dextrose) and nitrogen (yeast extract, soy protein isolate, and whey protein concentrate) sources were screened using one-variable-at-a-time and Plackett-Burman design, followed by Response Surface Methodology for optimization. Fermentation was scaled from a flask to 50 L fermenters at 37 °C and pH 6.50 ± 0.05. Cell viability, pH, and residual sugar were monitored. Functional assessments included stress tolerance assays (heat, acid, bile, and oxidative stress) and untargeted metabolomic profiling using ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry. The optimal medium comprised 9 g/L glucose, 14.1 g/L soy protein isolate, and 14.1 g/L yeast extract, supplemented with minerals. In 50 L fermentation, L22F achieved 9.20 log colony-forming units/mL at 12 h, with residual sugar at 1.50 g/L and pH 3.99. Compared to de Man, Rogosa, and Sharpe, the modified medium reduced production cost by 70%-88%, improved fermentation efficiency, and supported enhanced stress resilience. Metabolomic analysis revealed an elevated production of bioactive metabolites, particularly 1,4-dihydroxy-2-naphthoic acid and indolelactic acid, which are known to support gut homeostasis, anti-inflammatory responses, and probiotic efficacy. This study presents a cost-effective and scalable fermentation medium specifically designed for high-density L22F production. Beyond economic advantages, the medium enhanced the functional properties of L22F, supporting its application as a sustainable probiotic feed additive for swine. These findings establish a foundation for further industrial application and in vivo validation.

Coccidiosis, a parasitic disease caused by Eimeria species, remains a critical challenge for poultry production worldwide. Eimeria tenella is one of the most pathogenic species, causing intestinal lesions and compromising growth in chickens. This study aimed to estimate the prevalence of Eimeria species and evaluate the anticoccidial drug sensitivity of E. tenella field isolates in commercial broiler farms across Thailand. Four fecal samples were collected from each of the ten broiler farms in seven provinces of eastern and central Thailand. Oocysts were identified through morphological examination and confirmed through species-specific multiplex polymerase chain reaction. E. tenella isolates were propagated and subjected to in vivo anticoccidial sensitivity testing (AST) against four drugs: Nicarbazin (NIC), salinomycin (SAL), monensin (MON), and a combination of MON and NIC. Experimental infection trials were conducted on Ross 308 broiler chicks to assess weight gain, fecal oocyst shedding, lesion scores, and anticoccidial index (ACI) values. E. tenella and Eimeria praecox were the most prevalent species (40%), followed by Eimeria acervulina, Eimeria brunetti, and Eimeria mitis (20%). Mixed-species infections were detected in 50% of samples. The AST results showed that field isolates were sensitive to NIC, MON, and the MON + NIC combination, with ACI values of 172.51, 175.49, and 174.21, respectively. In contrast, SAL showed an ACI of 158.81, indicating resistance. All treated groups demonstrated reduced oocyst shedding and improved weight gain compared to untreated infected controls, though lesion score differences among treatments were not statistically significant. This study constitutes the first comprehensive report on anticoccidial drug resistance in E. tenella from broiler farms in Thailand. The findings indicate high efficacy of NIC, MON, and their combination, while revealing emerging resistance to SAL. These results provide crucial insights for revising coccidiosis control strategies and support the need for ongoing monitoring and development of alternative therapeutics to mitigate resistance evolution.

Cultural dietary practices involving the consumption of raw or undercooked meat, such as traditional sour fermented pork, pose significant risks for foodborne parasitic infections, particularly trichinellosis caused by Trichinella spiralis. This study aimed to evaluate the viability of T. spiralis larvae during sour pork fermentation and to assess the efficacy of microwave heating as a practical method for inactivating the larvae. Laboratory-bred hamsters were experimentally infected with T. spiralis to obtain encysted muscle larvae. Infected muscle samples were incorporated into a traditional sour pork recipe and fermented at ambient temperature (28-30°C) for 5 days. Larval viability was assessed daily using propidium iodide staining and confocal microscopy. In a separate experiment, pork slices embedded with infected muscle were subjected to microwave heating at 400 W (1-4 min) and 800 W (0.5-4.5 min). Post-treatment viability was determined similarly. Encysted larvae remained viable throughout the 5-day fermentation period, with no uptake of propidium iodide observed in any samples. In contrast, microwave heating at 400 W for 3 min or at 800 W for 1 min or longer resulted in complete larval inactivation, as evidenced by positive staining. Non-heated controls retained viable larvae, while boiling served as an effective positive control for inactivation. Traditional sour pork fermentation does not inactivate T. spiralis larvae within 5 days, underscoring a persistent zoonotic risk. However, microwave heating offers a rapid and accessible intervention for larval inactivation. These findings underscore the significance of public health education and food safety protocols in regions where the consumption of raw meat is culturally prevalent.

Feed cost constitutes a major constraint in small ruminant production systems in Jordan. The search for alternative, cost-effective feed ingredients has prompted interest in coconut meal (COC), a by-product of coconut oil extraction. This study evaluated the effects of including 10% COC (COC10) in the diet on the growth performance, nutrient utilization, carcass characteristics, and meat quality of growing Awassi lambs. Twenty-four male Awassi lambs (17.2 ± 0.63 kg) were randomly assigned to two dietary treatments: A control diet without COC (CON) and a test diet with 100 g/kg dry matter (DM) COC (COC10). The feeding trial lasted 70 days, comprising a 7-day adaptation and a 63-day experimental period. Feed intake and growth performance were monitored throughout. On day 49, five lambs per group were used to assess nutrient digestibility and nitrogen balance using total fecal and urine collection in metabolic cages. On day 64, lambs were slaughtered to evaluate carcass characteristics and meat quality. Blood samples were collected for serum biochemical analysis. The inclusion of COC10 significantly increased acid detergent fiber and ether extract intake (p ≤ 0.05), with no adverse effects on DM intake, weight gain, feed efficiency, or nutrient digestibility (p > 0.05). Economic analysis revealed a 16% reduction in production cost per kg of weight gain in the COC10 group (p = 0.05). No significant effects were observed on carcass traits, meat quality parameters, or blood biochemical profiles. Incorporating COC10 into the diet of Awassi lambs had no detrimental impact on growth, carcass characteristics, or health status, while improving economic efficiency. COC is a viable alternative protein and energy source in lamb diets. Further research is warranted to determine optimal inclusion rates under varying production systems.

Background and Aim:Neonatal piglet diarrhea (NPD) remains a significant challenge in the swine industry, contributing to elevated pre-weaning mortality, reduced productivity, and increased economic losses. In Vietnam, despite the growing importance of commercial pig production, comprehensive studies investigating the epidemiology of NPD and associated bacterial pathogens are lacking. This study aimed to assess the prevalence of Escherichia coli, Clostridium perfringens, and Clostridioides difficile and to identify risk factors contributing to NPD on Vietnamese sow farms.Materials and Methods:A cross-sectional study was conducted on 40 commercial sow farms across North, Central, and South Vietnam between August and December 2023. Structured questionnaires captured data on farm characteristics, management practices, and health interventions. Fecal samples from symptomatic piglets aged 1–14 days were pooled and analyzed using multiplex quantitative polymerase chain reaction to detect virulence genes of E. coli (F4, F5, F6, LT), C. perfringens (alpha, beta, and epsilon toxins), and C. difficile (toxins A and B). Logistic and ordinal regression models were applied to assess associations between risk factors and pathogen prevalence.Results:All farms tested positive for at least one pathogen. C. perfringens was the most prevalent (97.37%), followed by E. coli (46.49%) and C. difficile (39.47%). Co-infections involving multiple pathogens were common (64.91%), with C. perfringens consistently present in all mixed infections. Key virulence genes detected included LT (35.96%), alpha toxin (95.61%), and toxin A (20.17%). Significant risk factors included farm type, region, weaning performance, and peripartum antibiotic administration route. Notably, farms using mixed-feed antibiotics exhibited higher E. coli prevalence. Larger farms and those practicing early piglet relocation also showed increased pathogen diversity.Conclusion:This study presents the first national-scale assessment of bacterial pathogens in NPD across Vietnamese sow farms. The findings highlight the high burden of toxin-producing bacteria, frequent co-infections, and multiple farm-level risk factors. Interventions such as targeted vaccination, optimized antibiotic use, improved weaning practices, and enhanced regional surveillance are essential for mitigating NPD impacts and improving piglet health outcomes in Vietnam.