Issue Cover
Volume 18 | June
Research Article | 06 Jun 2025
Effects of probiotics and amprolium on performance, lesion scores, oocyst shedding, and histopathological changes in Eimeria tenella-infected broiler chickens
Thanyakorn Chalalai, Watcharapon Promsut, Kannika Hinkhao, Tirocha Hengphrathani, Kamonporn Sangsakul, Nopparat Bhavabhutanon, and Tippayaporn Nonkookhetkhong

Volume-18 | Issue-6 | Article-1 | https://doi.org/10.14202/vetworld.2025.1400-1410

Preview Abstract
Coccidiosis caused by Eimeria tenella significantly impairs poultry health and productivity, prompting the search for alternative or complementary therapies to conventional coccidiostats. This study investigates the prophylactic and therapeutic efficacy of a probiotic complex of Lactobacillus, Bifidobacteria, Enterococcus, and Streptococcus, alone or in combination with amprolium, against E. tenella infection in broiler chickens. A total of 90 broiler chickens were randomly allocated into six experimental groups (n = 15/group). Group 1 served as the uninfected control, while Group 2 comprised infected but untreated controls. Group 3 received probiotics at a concentration of 1 mg/mL, and Group 4 was treated with amprolium at a dosage of 20 mg/kg. Group 5 was administered a combination of probiotics (1 mg/mL) and amprolium (20 mg/kg), whereas Group 6 received prophylactic treatment with probiotics (1 mg/mL). All groups except the uninfected control were challenged orally with 2 × 104 sporula-ted E. tenella oocysts. Growth performance was monitored on days 15, 21, and 28. Lesion scoring, oocyst shedding, and histopathological examinations were conducted on day 28. An in vitro sporulation assay evaluated the inhibitory potential of treatments on oocyst development. In vitro, the probiotic-amprolium combination significantly reduced oocyst sporulation rates (5.86%). In vivo, amprolium and prophylactic probiotics significantly improved body weight gain and feed intake (p < 0.05) and reduced oocyst shedding. Lesion severity and parasite stage counts were significantly lower in the amprolium group; however, the combination group exhibited unexpectedly higher lesion scores. Mortality was highest in the amprolium and untreated groups (20%) but absent in the combination group, suggesting differential immunomodulatory effects. Histopathological analysis confirmed reduced intestinal damage in groups treated with amprolium or prophylactic probiotics. Amprolium monotherapy and prophylactic probiotic supplementation were effective in mitigating E. tenella-induced pathology and improving broiler performance. Probiotics alone provided moderate benefits, while their post-infection therapeutic use or co-administration with amprolium did not yield superior results. These findings underscore the prophylactic value of probiotics and warrant further studies to optimize combination regimens under field conditions.

Volume-18 | Issue-6 | Article-2 | https://doi.org/10.14202/vetworld.2025.1411-1420

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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.

Volume-18 | Issue-6 | Article-3 | https://doi.org/10.14202/vetworld.2025.1421-1432

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The rising burden of antimicrobial resistance (AMR) in veterinary medicine poses significant threatsto animal and public health. In South Africa, inadequate surveillance exacerbates the challenge, particularly regardingStaphylococcus spp. infections in companion animals. This study aimed to investigate the patterns and predictors of AMRand multidrug resistance (MDR) in Staphylococcus isolated from dogs between 2012 and 2017. A retrospective cross-sectional study was conducted on 1627 Staphylococcus isolates. Dataregarding animal demographics and antimicrobial susceptibility were extracted, cleaned, and analyzed. Intermediatesusceptibility results were classified as resistant. AMR was defined as resistance to at least one antimicrobial from oneclass and MDR as resistance to antimicrobials from three or more classes. Descriptive statistics, Cochran–Armitagetrend analysis, and binary logistic regression models were employed to assess trends and predictors of AMR and MDR. Overall, 61.2% of isolates exhibited resistance to at least one antimicrobial, and 39.0% were classified as MDR. Thehighest resistance was observed against penicillins (39.64%), followed by aminoglycosides (22.31%). Significant predictorsof AMR included Staphylococcus species, specimen type, and year of isolation, while MDR was significantly associated withspecimen type and the age of the dog. Notably, Staphylococcus pseudintermedius showed a markedly higher likelihood ofresistance (Adjusted Odds Ratio = 2.23, p < 0.001) compared to other species. Temporal trends indicated a decrease in AMRbut an increase in MDR across the study period. The high prevalence of AMR and MDR among canine Staphylococcus isolates, particularly in skin infections andamong younger dogs, underscores the urgent need to strengthen antimicrobial stewardship, enhance surveillance systems,and target interventions in veterinary practice. These findings serve as critical baseline data for future assessments of AMRtrends and can be used to inform strategies to mitigate the dissemination of resistant pathogens between animals andhumans.
Research Article | 06 Jun 2025
Reproductive impacts and disease burden of metritis and placental retention in dairy cows: A longitudinal monitoring study in Southern Vietnam (2022–2024)
Thuong Thi Nguyen, Lien Thi Bich Nguyen, Khang Nguyen Duong, and Thuan Khanh Nguyen

Volume-18 | Issue-6 | Article-4 | https://doi.org/10.14202/vetworld.2025.1433-1439

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Postpartum reproductive disorders, particularly metritis and placental retention, significantly compromise dairy herd productivity and calf viability. In Vietnam, where dairy production is expanding, limited data exist on the prevalence and consequences of such disorders under modern farm conditions. This study aimed to determine the prevalence of metritis and placental retention in dairy cows on an industrial farm in southern Vietnam from 2022 to mid-2024 and to evaluate their reproductive and neonatal health consequences. A longitudinal study was conducted on a dairy herd monitored through skin conductance responses activity sensors and DataFlow™ II software (Allflex Livestock Intelligence, MSD Animal Health Intelligence, USA). Health alerts triggered clinical examinations to identify genital infections. Metritic cows were treated using antibiotics and hormonal therapies and were monitored for recovery and subsequent breeding success. Neonatal calves were examined for respiratory and gastrointestinal disorders for 21 days post-birth. Statistical analysis was performed using Chi-square tests at a 95% confidence level. Genital infections affected 23.5%–38.8% of cows annually, with metritis prevalence ranging from 7.2% to 9.8%. Placental retention remained consistent at approximately 13% across years. Quarter 2 consistently exhibited the highest incidence of reproductive disorders. Treatment success for metritis was high (85.8%–88.6%); however, post-treatment pregnancy rates declined over time (68.5% in 2022 and 54.8% in 2024). Neonatal respiratory infections (2.9%–4.4%) were more frequent than gastrointestinal infections (0.1%–0.8%), with calf mortality declining from 3.4% in 2022 to 0.7% in 2024. Metritis and placental retention remain prevalent challenges in Vietnamese dairy herds, adversely impacting reproductive efficiency and calf health despite high treatment efficacy. The seasonal spike in disease incidence underscores the need for tailored herd health management during hotter months. Although early detection through precision monitoring improved recovery outcomes, residual effects on fertility persisted. Strengthened periparturient care, postpartum surveillance, and colostrum management are recommended to enhance both maternal and neonatal health outcomes.
Systematic Review | 06 Jun 2025
Global prevalence and risk factors of equine infectious anemia: A systematic review and meta-analysis
Lintang Winantya Firdausy, Faisal Fikri, Arya Pradana Wicaksono, Hakan Çalışkan, and Muhammad Thohawi Elziyad Purnama

Volume-18 | Issue-6 | Article-5 | https://doi.org/10.14202/vetworld.2025.1440-1451

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Equine infectious anemia (EIA) is a lentiviral disease affecting members of the Equidae family, with global distribution and significant implications for animal health and biosecurity. Despite numerous individual reports, a comprehensive synthesis of its global prevalence and risk factors remains lacking. This study aimed to conduct a systematic review and meta-analysis to estimate the global prevalence of EIA, identify diagnostic trends, and evaluate factors associated with heterogeneity across studies. A systematic search was conducted in six major databases (PubMed, Scopus, Web of Science, ScienceDirect, Cochrane Library, and ProQuest), yielding 312 records. After Preferred Reporting Items for Systematic Reviews and Meta-Analyses-guided screening, 29 eligible studies published between 1975 and 2024 were included in the study. Meta-analysis was performed using R Studio (version 4.4.2) employing a random-effects model. Subgroup analyses and meta-regression were conducted to explore heterogeneity across host species, continent, diagnostic method, and study period. Publication bias was assessed through funnel plots and Egger’s test. The global pooled prevalence of EIA was estimated at 20.97% (95% confidence interval [CI]: 11.08–30.85), with substantial heterogeneity (I2 = 99.3%). South America reported the highest regional prevalence (27.21%), while horses showed the greatest susceptibility among Equidae (25.40%). Diagnostic methods varied, with agar gel immunodiffusion being the most commonly used (18.62% prevalence detection). A declining trend in prevalence (2.19%–28.70%) was noted from 2015 to 2022. No significant publication bias was detected. Meta-regression revealed that climate and study period partially explained the heterogeneity. This study highlights the substantial global burden and diagnostic variability of EIA, emphasizing the need for enhanced surveillance in endemic areas, standardized diagnostic protocols, and strengthened quarantine practices. Expanding serological monitoring in underrepresented regions and integrating climatic and ecological data into control strategies are vital for mitigating EIA transmission risks.

Volume-18 | Issue-6 | Article-6 | https://doi.org/10.14202/vetworld.2025.1452-1465

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Despite the profound cultural and ecological significance of falcons in the Arabian Gulf, systematic evaluations of regional falcon research are lacking. A bibliometric approach can elucidate the evolution, influence, and emerging priorities within this specialized field. This study aimed to provide the first comprehensive bibliometric mapping of falcon research in the Arabian Gulf countries over the past four decades (1984–2024), revealing research dynamics, international collaboration networks, thematic trajectories, and critical knowledge gaps. Original research articles were systematically retrieved from the Scopus database using targeted search strategies restricted to title-level keywords and affiliation filters. Following Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, 126 articles were selected for analysis. Bibliometric methods – including co-authorship networks, keyword co-occurrence, thematic mapping, and trend analysis – were applied using VOSviewer, Bibliometrix, and Excel. The research output exhibited a modest annual growth rate of 2.32%, predominantly driven by contributions from the United Arab Emirates and Saudi Arabia. Strong international collaborations (56.35% co-authored publications) were identified, particularly with European and North American institutions. Research themes historically centered on clinical veterinary topics and avian biology are now shifting toward molecular diagnostics, genetic studies, and disease surveillance. However, a clear regional imbalance and underrepresentation of emerging fields such as genomics, epidemiology, and conservation breeding were observed. This analysis underscores the need for an integrative, regionally inclusive research strategy incorporating advanced molecular technologies and conservation science. Strengthening cross-border collaboration, adopting genome-based monitoring, and addressing emerging infectious threats will be critical to advancing falcon research and preservation in the Arabian Gulf.
Research Article | 10 Jun 2025
Propolis mitigates histopathological alterations in the pituitary gland and reproductive system of female albino rats subjected to cadmium toxicity
Abdulla A. Albishtue, Aqeel Mohsin Al-Mahmmodi, Hasan A. Almamoori, and Mustafa Ali Alahmer

Volume-18 | Issue-6 | Article-7 | https://doi.org/10.14202/vetworld.2025.1466-1478

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Cadmium (Cd) is a pervasive environmental toxin that disrupts endocrine function and induces oxidative damage in reproductive organs. Propolis (PRO), a resinous substance produced by bees, has garnered attention for its antioxidant and estrogenic properties. This study investigated the protective potential of PRO on the pituitary-ovarian-uterine axis in female rats subjected to Cd-induced toxicity. Thirty adult female albino rats were randomized into five groups (n = 6/group): Control (C), Cd-only (T0), and Cd plus PRO at 150, 300, and 500 mg/kg body weight (BW) (T1–T3, respectively). Cadmium chloride was administered orally at 5 mg/kg for 4 weeks. PRO was co-administered daily through gavage. At the proestrus stage, animals were euthanized for tissue collection. Vaginal cytology was used to confirm estrous stage. Histopathological examination of the ovary, uterus, and pituitary gland was performed using H&E staining. Serum estradiol (E2) and superoxide dismutase (SOD) activity were assessed to evaluate hormonal and oxidative responses. Morphometric measurements were statistically analyzed through one-way analysis of variance with Tukey’s post hoc test. Cd exposure (T0) led to prolonged estrous cycles, ovarian atresia, uterine degeneration, and significant disruption of pituitary architecture, accompanied by reduced E2 and SOD levels (p < 0.05). PRO administration dose-dependently ameliorated these alterations. The highest PRO dose (T3) restored the histological architecture of all target organs to near-normal levels, significantly increased ovarian and uterine weight ratios, and elevated both E2 and SOD activity. Histomorphometric analysis confirmed increased follicle survival, thickened ovarian surface epithelium, and elevated interstitial cell counts. Pituitary endocrine cell counts and uterine gland numbers were also significantly higher in PRO-treated groups, particularly T3. PRO supplementation at 500 mg/kg BW significantly attenuates Cd-induced reproductive and endocrine toxicity in female rats by restoring histological integrity and enhancing antioxidant and estrogenic responses. These findings suggest PRO as a promising candidate for mitigating heavy metal-induced reproductive dysfunction.
Research Article | 10 Jun 2025
Effect of idebenone supplementation in a semen extender on boar spermatozoa quality during liquid storage
Rehardus Ricco Pantecostoma, Jatesada Jiwakanon, and Saksiri Sirisathien

Volume-18 | Issue-6 | Article-8 | https://doi.org/10.14202/vetworld.2025.1479-1486

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In swine production, over 99% of artificial insemination (AI) procedures utilize boar semen preserved in liquid form at 17°C for up to 5 days. However, spermatozoa are highly susceptible to oxidative stress during storage, which impairs motility, membrane integrity, and overall fertility. Reactive oxygen species-induced lipid peroxidation (LPO) compromises sperm structure and function. Although antioxidants are used to mitigate oxidative damage, idebenone (IDB) – a short-chain benzoquinone with potent mitochondrial antioxidant properties – has not been studied in boar semen preservation. This study aimed to investigate the effects of IDB supplementation in a semen extender on boar spermatozoa quality during 120 h of liquid storage at 17°C. Ejaculates from 25 Duroc boars were diluted with Beltsville Thawing Solution to a final concentration of 30 × 106 sperm/mL. In Experiment 1, semen samples (n = 13) were treated with 0, 78, 156, 312, 625, or 1250 nM of IDB and assessed at 24-h intervals for motility (computer-assisted sperm analysis), viability (eosin-nigrosin), and membrane integrity (hypo-osmotic swelling test). In Experiment 2, 78 nM IDB was selected for its optimal effects and further tested (n = 12) for acrosome integrity (fluorescein isothiocyanate-conjugated peanut agglutinin), capacitation status (chlortetracycline staining), LPO (thiobarbituric acid reactive substances), and kinematic parameters. After 120 h, the 78 nM IDB group showed significantly improved total motility (81.4%) and membrane integrity (60.3%) compared to control (72.9% and 46.7%, respectively; p < 0.05). Viability was higher in IDB-treated groups, and high concentrations (625–1250 nM) reduced motility. In Experiment 2, 78 nM IDB improved total and progressive motility (84.0% and 46.8%), preserved acrosome integrity (64.06% vs. 58.75%), reduced capacitated sperm (64.72% vs. 59.11%), and decreased malondialdehyde levels (51.84 nmol vs. 59.61 nmol) relative to controls (p < 0.05). IDB at 78 nM effectively preserves boar semen quality during 5-day liquid storage by enhancing motility, viability, membrane and acrosome integrity, and reducing oxidative stress. These findings highlight its potential as a novel antioxidant additive in AI protocols.
Research Article | 10 Jun 2025
Gut microbiota modulation and immunity enhancement by Bacillus amyloliquefaciens NL1.2: A fiber-degrading probiotic isolated from native Thai swine
Kittiya Khongkool, Malai Taweechotipatr, Sunchai Payungporn, Vorthon Sawaswong, and Monthon Lertworapreecha

Volume-18 | Issue-6 | Article-9 | https://doi.org/10.14202/vetworld.2025.1487-1507

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The pursuit of sustainable alternatives to antibiotic growth promoters has intensified interest in spore-forming probiotics with fiber-degrading capabilities. This study aimed to isolate, characterize, and evaluate the safety and functional properties of Bacillus spp. from native Thai swine, focusing on strains with probiotic potential and enzymatic activity for application in livestock nutrition. Spore-forming Bacillus isolates were obtained from fecal samples of backyard-raised native pigs. Isolates were screened for acid and bile tolerance, autoaggregation, hydrophobicity, biofilm formation, adhesion to Caco-2 cells, antimicrobial activity, and co-aggregation with pathogens. Enzyme production (cellulase, xylanase, and pectinase), hemolytic activity, and antibiotic susceptibility were also assessed. The most promising strain, Bacillus amyloliquefaciens NL1.2, was subjected to in vivo safety and efficacy evaluations in a mouse model, including assessments of toxicity, histopathology, secretory immunoglobulin A (IgA) levels, and gut microbiome modulation through full-length 16S ribosomal RNA sequencing. B. amyloliquefaciens NL1.2 exhibited robust probiotic traits including high acid (115.05%) and bile (75.16%) tolerance, strong autoaggregation (65.99%), moderate hydrophobicity (34.13%), and effective adhesion (2.0%) to intestinal epithelial cells. It produced fiber-degrading enzymes (cellulase: 0.015 U/mL; xylanase: 0.522 U/mL; and pectinase: 0.374 U/mL) showed antimicrobial activity against Enterohemorrhagic Escherichia coli, Enteropathogenic E. coli, and Salmonella Typhimurium, and was non-hemolytic and antibiotic-sensitive. In vivo, NL1.2 induced no adverse effects and significantly elevated intestinal secretory IgA levels (p < 0.05). Microbiome analysis revealed enrichment of beneficial taxa (e.g., Bacteroidetes and Barnesiella) and reduction of potentially pathogenic taxa (e.g., Helicobacter and Deferribacteres). B. amyloliquefaciens NL1.2 is a safe, multifunctional probiotic with fiber-degrading, immunomodulatory, and gut microbiota-modulating properties. Its origin from native swine and broad functional attributes highlights its potential as a next-generation feed additive for sustainable animal production.
Research Article | 15 Jun 2025
Managing African animal trypanosomiasis in Benin: Knowledge, attitudes, and practices of cattle owners in the West Atacora zone
Yao Akpo, Aretas B. N. Tonouhewa, Traore Alkoiret, and Marc T. Kpodekon

Volume-18 | Issue-6 | Article-10 | https://doi.org/10.14202/vetworld.2025.1508-1516

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African animal trypanosomiasis (AAT), transmitted by tsetse flies, severely constrains livestock productivity in sub-Saharan Africa. In Benin, limited governmental control initiatives and widespread drug misuse have raised concerns about emerging trypanocidal resistance. This study aimed to assess the knowledge, attitudes, and practices of cattle farmers in the Atacora and Donga departments of northern Benin and identify behaviors contributing to the persistence and drug resistance of AAT. A cross-sectional survey was conducted from September to December 2024 among 201 cattle farmers selected through stratified random sampling across five districts. Data were collected using a semi-structured, pre-tested questionnaire and analyzed with R software employing descriptive statistics and comparative tests (χ2, Mann–Whitney U, and Kruskal–Wallis). Awareness of tsetse flies and their role in AAT transmission was high (84%), but only 24% recognized insecticide-based vector control as an effective method. The predominant control strategy involved trypanocides – mainly diminazene aceturate (81%) and isometamidium chloride (71%) – with 99% of participants administering these drugs. Notably, 42% sourced trypanocides from illicit markets, and 22% practiced self-medication. Most farmers (56%) treated their cattle twice yearly, yet 65% failed to observe withdrawal periods, and 59% reported therapeutic failures. Only 12% had received formal training in AAT management. Despite high disease awareness, poor adherence to recommended control practices and the prevalent misuse of trypanocides, particularly through informal markets, pose serious threats to sustainable AAT management. There is an urgent need for integrated risk communication and policy-driven interventions promoting responsible drug use and vector control in northern Benin.
Research Article | 15 Jun 2025
Enhancing the nutritional profile of African sharptooth catfish (Clarias gariepinus) through dietary supplementation with natural minerals and probiotic Escherichia coli 39-SN
Nurzhan Sarsembayeva, Gulmariya Ikramzhan, Tolkyn Abdigaliyeva, Zhumagul Kirkimbaeva, Birzhan Biyashev, Saule Sherimova, and Primkul Ibragimov

Volume-18 | Issue-6 | Article-11 | https://doi.org/10.14202/vetworld.2025.1517-1526

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Feed additives composed of natural minerals and probiotics are increasingly explored in aquaculture to improve fish health and meat quality while reducing antibiotic dependency. Zeolite and vermiculite, due to their adsorptive and ion-exchange properties, enhance mineral bioavailability, whereas probiotics, such as Escherichia coli 39-SN improve digestive function and nutrient assimilation. This study aimed to evaluate the effects of dietary inclusion of zeolite or vermiculite in combination with E. coli 39-SN on the chemical, mineral, and amino acid composition of African sharptooth catfish (Clarias gariepinus). A total of 150 catfish were divided into three groups: A control group fed a standard diet, and two experimental groups receiving diets supplemented with either 5% zeolite or 5% vermiculite, each combined with 0.1% E. coli 39-SN. The trial lasted 87 days under controlled aquaculture conditions. Post-trial, muscle tissues were analyzed for moisture, fat, protein, ash content, energy value, mineral content (e.g., iron, zinc, and magnesium), and amino acid profiles using standardized laboratory methods. Fish in the experimental groups exhibited improved nutritional profiles compared to the control. Notably, the zeolite + probiotic group showed the highest total amino acid content (34.63%) and significant increases in essential amino acids, such as leucine and phenylalanine. Vermiculite + probiotic supplementation yielded the highest fat content (0.69 g/100 g) and increased concentrations of iron and zinc. Both experimental groups demonstrated improved energy values, elevated mineral levels, and enhanced biological value of the muscle tissue. No signs of stress or cannibalism were observed, indicating good tolerance to the diets. The synergistic use of natural minerals and E. coli 39-SN significantly enhanced the chemical and nutritional quality of C. gariepinus muscle tissue. These findings suggest that such dietary strategies offer a promising alternative to antibiotics and can enhance the commercial value of aquaculture products. Future studies should explore dose optimization, long-term health effects, and the immunomodulatory potential of E. coli 39-SN in fish.
Research Article | 15 Jun 2025
Prevalence and characterization of gastrointestinal and ectoparasites in long-tailed macaques (Macaca fascicularis) from ecotourism regions of Aceh, Indonesia
Muhammad Hanafiah, Teuku Reza Ferasyi, Erdiansyah Rahmi, Winaruddin Winaruddin, Kartika Dewi, Roliamy Saputri, and Nisrima Redukmi

Volume-18 | Issue-6 | Article-12 | https://doi.org/10.14202/vetworld.2025.1527-1539

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Long-tailed macaques (Macaca fascicularis) serve as critical sentinels for zoonotic disease surveillance due to their ecological proximity to human populations. Understanding their parasitic burden is vital for conservation and public health, particularly in ecotourism areas where human-primate interactions are frequent. This study aimed to investigate the prevalence, diversity, and morphological characteristics of gastrointestinal (GI) and ectoparasites in M. fascicularis across four natural habitats in Aceh Province, Indonesia. A total of 100 fecal samples were collected from wild macaques at four sites: Pulau Weh Sabang Nature Tourism Park, Kuala Langsa Mangrove Forest, Saree (Aceh Besar), and Aceh Jaya. The parasitological examination involved the centrifugation method, lactophenol staining, and scanning electron microscopy (SEM). Parasites were identified based on egg morphology and adult worm anatomy. Of the 100 samples analyzed, 45% tested positive for GI parasites. Nematode prevalence was highest (80%), followed by protozoa (10%) and ectoparasites (10%). Identified nematodes included Ancylostoma spp. (70%), Oesophagostomum spp. (50%), Strongyloides spp. (40%), Ascaris spp. (30%), Enterobius spp. (20%), and Trichuris spp. (10%). Protozoan (Balantidium spp.) and ectoparasitic (Psoroptes spp.) infections were less common. Mixed infections were more frequent (70%) than single infections (30%). Adult worms examined through SEM and lactophenol staining were confirmed to be two nematode species: Oesophagostomum (Conoweberia) bifurcum and Trichuris trichiura, both with zoonotic potential. The high prevalence of nematodes, particularly zoonotic species, underscores the importance of monitoring parasitic infections in M. fascicularis residing in tourist-exposed areas. This study highlights the utility of combining traditional and advanced diagnostic techniques to enhance parasite surveillance. Integrating health assessments of wild primate populations into conservation programs is recommended to mitigate zoonotic risk and support One Health objectives.
Research Article | 15 Jun 2025
Evaluation of serum alpha-1B glycoprotein and C-reactive protein levels as biomarkers of canine benign prostatic hyperplasia
Grisnarong Wongbandue, Sekkarin Ploypetch, Piyada Pruksakitcharoen, Kittithee Udomrit, Kanisa Nujan, Rinrada Seekhumtae, Tanapron Thubthim, and Nawarus Prapaiwan

Volume-18 | Issue-6 | Article-13 | https://doi.org/10.14202/vetworld.2025.1540-1548

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Benign prostatic hyperplasia (BPH) is a prevalent disorder in aging male dogs, characterized by prostate enlargement secondary to hormonal dysregulation and chronic inflammation. Identifying non-invasive biomarkers is crucial for improving diagnosis and monitoring therapeutic interventions. This study aimed to evaluate serum alpha-1B glycoprotein (A1BG) and C-reactive protein (CRP) concentrations in dogs with BPH before and after castration, to assess their diagnostic and prognostic utility. A total of 20 male dogs were assigned to two groups: healthy controls (n = 10) and BPH-affected dogs (n = 10). Blood samples were collected from controls and the BPH group at diagnosis and 1 month post-castration. Serum A1BG and CRP concentrations were measured using enzyme-linked immunosorbent assay and fluorescence immunoassay, respectively. Prostatic volume (PV) was evaluated ultrasonographically. Dogs with BPH demonstrated significantly lower serum A1BG concentrations before castration compared to healthy controls (p < 0.01) and post-castration (p < 0.01). Post-castration A1BG levels were comparable to controls, suggesting biochemical normalization. Serum CRP concentrations remained within the normal range (<30 mg/L) across all groups and showed no significant differences. A significant negative correlation was observed between age and A1BG concentration in the pre-castration BPH group (r = −0.74, p = 0.02). Castration resulted in a marked reduction in PV, consistent with therapeutic response. Serum A1BG demonstrated potential as a sensitive biomarker for the diagnosis and therapeutic monitoring of canine BPH, in contrast to CRP, which exhibited limited diagnostic value. Normalization of A1BG levels post-castration supports its role in reflecting disease resolution. Integrating A1BG assessment into veterinary diagnostic workflows could enhance early detection, monitoring, and management strategies for BPH, offering a non-invasive and clinically informative approach. Further longitudinal studies with larger cohorts are warranted to validate these findings and explore long-term biomarker dynamics.
Research Article | 15 Jun 2025
Molecular epidemiology and antimicrobial resistance of Salmonella at the human–macaque–environment interface in Thailand: A One Health surveillance study
Suchawan Pornsukarom, Daraka Tongthainan, Phairot Phromwat, Suwarak Wannaratana, Kulchai Nakbubpa, and Sarut Muangsri

Volume-18 | Issue-6 | Article-14 | https://doi.org/10.14202/vetworld.2025.1549-1560

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The close interaction between humans and free-ranging macaques in urbanized environments raises concerns about the potential transmission of antimicrobial-resistant zoonotic pathogens. This study applied a One Health approach to estimate the prevalence, serovar distribution, antimicrobial resistance (AMR), and genetic diversity of Salmonella spp. in long-tailed macaques (Macaca fascicularis) and environmental sources in Chonburi, Thailand. A total of 313 samples – including 224 rectal swabs from macaques and 89 environmental samples (pooled macaque feces, stray dog feces, soil, feed, and drain water) – were collected from Si Racha and Sattahip districts between April and July 2023. Salmonella isolation was conducted using conventional culture methods, followed by confirmation through serotyping and polymerase chain reaction targeting the invA gene. Antimicrobial susceptibility testing was performed against 14 agents using broth microdilution. Multi-locus sequence typing and 16S rRNA gene sequencing were conducted to assess phylogenetic diversity. The overall prevalence of Salmonella was 2.88%, with all positive samples detected in the Si Racha district. Environmental samples had a significantly higher prevalence (8.89%) than macaque rectal swabs (0.45%; odds ratio = 22; 95% confidence interval: 2.71–178.84; p = 0.0002). Six distinct serovars were identified, with Salmonella Corvallis predominating in macaque feces. Among the nine isolates, 77.78% exhibited resistance, primarily to tetracycline and ampicillin. Notably, 85.71% of AMR strains from environmental samples were multidrug-resistant (MDR), showing resistance to ≥6 antimicrobials. Phylogenetic analysis revealed genetic heterogeneity, with no clear clustering by source or serovar. This study underscores the circulation of MDR Salmonella within macaques and their surrounding environments, implicating environmental reservoirs in potential zoonotic and reverse zoonotic transmission. The findings advocate for public awareness initiatives, environmental hygiene improvements, and integrative One Health strategies to mitigate AMR dissemination at the human–animal–ecosystem interface.
Research Article | 15 Jun 2025
Prevalence and anticoccidial drug sensitivity of Eimeria tenella isolated from commercial broiler farms in Thailand
Suttitas Tongkamsai, Siraprapa Boobphahom, Rachan Apphaicha, and Niwat Chansiripornchai

Volume-18 | Issue-6 | Article-15 | https://doi.org/10.14202/vetworld.2025.1561-1570

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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.
Research Article | 16 Jun 2025
Genome-wide characterization and comparative phylogenomics of three Salmonella Abortusequi strains isolated from equine abortions in Kazakhstan
Temirlan Bakishev, Asylulan Amirgazin, Marat Kuibagarov, Alexander Shevtsov, Zhanar Bakisheva, Gulzhan Yessembekova, Alma Kairzhanova, Ablaikhan Kadyrov, Kui Guo, Xiaojun Wang, Sarsenbay Abdrakhmanov, and Sergey Borovikov

Volume-18 | Issue-6 | Article-16 | https://doi.org/10.14202/vetworld.2025.1571-1580

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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.
Research Article | 16 Jun 2025
Impact of apolipoprotein B-associated cholesterol deficiency genotype on milk composition, somatic cell count, and parity effects in Lithuanian Holstein cows
Ramutė Mišeikienė, Nijolė Pečiulaitienė, Lina Kajokienė, Renata Bižienė, Kristina Morkūnienė, Vilius Marma, Saulius Tušas, Paulius Matusevičius, Ewa Wójcik, Alina Janocha, Anna Milczarek, and Laimutis Kučinskas

Volume-18 | Issue-6 | Article-17 | https://doi.org/10.14202/vetworld.2025.1581-1589

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Cholesterol deficiency (CD) in Holstein cattle, caused by a loss-of-function mutation in the apolipoprotein B (APOB) gene, is a heritable autosomal recessive condition with known implications for fat metabolism and cholesterol transport. This study aimed to investigate the effect of the CD genotype on milk yield components, cholesterol concentration, and somatic cell count (SCC) in Lithuanian Holstein cows, and to determine whether lactation number modulates these relationships. A total of 188 cows were classified by lactation: 1st (n = 44), 2nd (n = 50), 3rd and 4th (n = 60), and ≥5th (n = 34). Genotyping for the APOB mutation was conducted using allele-specific polymerase chain reaction. Milk fat, protein, lactose, and SCC were determined using LactoScope Fourier-transform infrared spectroscopy and Somascope methods, while cholesterol concentration was measured by high-performance liquid chromatography. Statistical analysis involved the Kruskal–Wallis H test due to non-normal data distribution. The heterozygous CD genotype was identified in 17.02% of the population, with wild-type and mutant allele frequencies of 0.91 and 0.09, respectively. Non-carriers showed marginally higher fat, protein, and cholesterol levels, with a statistically significant difference in fat content (p = 0.04). When stratified by lactation, significant differences were observed for fat content in the 1st lactation group (p = 0.026), SCC in the 2nd (p = 0.038), and protein content in the 3rd (p = 0.030). No significant variation in milk cholesterol concentration was detected across genotype groups in any lactation group. This study confirms the presence of the CD-associated APOB allele in the Lithuanian Holstein population. While CD status significantly influenced milk fat percentage, its effect on other milk composition traits and SCC was limited. Parity exhibited specific but non-consistent modulating effects. Further large-scale, longitudinal studies are warranted to elucidate the physiological underpinnings of these findings.
Research Article | 16 Jun 2025
Co-circulation and genetic characterization of genotype I and II feline bocavirus strains in domestic cats from Northern Vietnam
Hieu Van Dong, Giang Thi Huong Tran, Yen Hoang Thi Nguyen, Thiet Chi Ngo, Amonpun Rattanasrisomporn, Chaiwat Boonkaewwan, Dao Anh Tran Bui, and Jatuporn Rattanasrisomporn

Volume-18 | Issue-6 | Article-18 | https://doi.org/10.14202/vetworld.2025.1590-1598

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Feline bocavirus (FBoV), a member of the Parvoviridae family, has been implicated in gastrointestinal and respiratory conditions in domestic cats. Despite increasing global recognition, the molecular epidemiology of FBoV in Vietnamese animal populations remains largely unexplored. This study aimed to detect and genetically characterize FBoV strains circulating among domestic cats in Northern Vietnam to better understand their genotypic diversity and potential clinical relevance. A total of 166 fecal samples were collected from domestic cats of varying age, sex, and clinical status across four provinces in Northern Vietnam between 2022 and 2023. DNA was extracted and screened for FBoV using conventional polymerase chain reaction targeting the non-structural (NS)-1 gene. Positive samples were subjected to Sanger sequencing, and partial NS1 sequences were analyzed using MEGA X for phylogenetic inference. Recombination analysis was performed using RDP 4.0, and statistical significance was assessed using Fisher’s exact test. FBoV DNA was detected in 4 of 166 samples (2.41%), including one from a diarrheic cat and three from healthy cats. Phylogenetic analysis of the partial NS1 gene revealed that three strains belonged to genotype I and one to genotype II, all showing close genetic similarity to Chinese strains. Nucleotide identities among Vietnamese strains ranged from 64.68% to 99.57%. No recombination events were observed among the detected strains. FBoV was detected across age groups and both sexes, without significant associations. Co-infections with other enteric viruses (feline coronavirus, feline panleukopenia virus, feline astrovirus, and feline kobuvirus) were not observed in the FBoV-positive samples. This study provides the first molecular evidence of co-circulating FBoV genotypes I and II in domestic cats in Vietnam, indicating viral genetic diversity and suggesting possible regional transmission routes linked to neighboring countries. While FBoV was present in both symptomatic and asymptomatic cats, its clinical significance remains inconclusive. The findings underscore the need for expanded surveillance, complete genome analyses, and investigation into FBoV’s pathogenic potential and co-infection dynamics in the feline population. These data will be instrumental in shaping future diagnostic and control strategies for feline viral enteritis in Vietnam.
Review Article | 16 Jun 2025
African swine fever virus: Virology, pathogenesis, clinical impact, and global control strategies
Tridiganita Intan Solikhah, Firda Rostiani, Assyuria Fahma Putri Nanra, Adilah Dwi Putri Paras Dewi, Putri Haibah Nurbadri, Qurrotul Aini Dwi Agustin, and Gahastanira Permata Solikhah

Volume-18 | Issue-6 | Article-19 | https://doi.org/10.14202/vetworld.2025.1599-1613

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African swine fever (ASF) is a highly contagious and lethal viral disease affecting domestic pigs and wild boars, with profound implications for global swine production and food security. Caused by the ASF virus (ASFV), a complex double-stranded DNA virus of the Asfarviridae family, the disease exhibits diverse clinical outcomes - from peracute death to chronic infection - depending on viral genotype and host immunity. ASFV primarily targets monocytes and macrophages, leading to severe lymphoid depletion, systemic inflammation, and vascular pathology mediated by cytokine storms. The virus demonstrates remarkable environmental resilience and is transmitted through direct contact, fomites, and biological vectors such as Ornithodoros soft ticks. With 23 genotypes identified to date, ASFV poses ongoing challenges to diagnosis, control, and vaccine development. Diagnostic methods, including polymerase chain reaction, enzyme-linked immunosorbent assay, and virus isolation, are essential for timely detection and containment. Despite advances in live-attenuated vaccine research, safe and broadly protective vaccines remain elusive. This review synthesizes current knowledge on ASFV’s molecular biology, transmission dynamics, immunopathogenesis, clinical presentations, and control strategies and underscores the urgent need for integrated surveillance systems, cross-sectoral collaboration, and innovative tools for outbreak prediction and disease mitigation.
Review Article | 19 Jun 2025
Reproductive integration of leptin and kisspeptin in small ruminants: Mechanisms, biomarker potential, and prospects for precision breeding
Herdis Herdis, Ismeth Inounu, Santoso Santoso, Rahma Isartina Anwar, Sari Yanti Hayanti, Mohammad Firdaus Hudaya, Desiana Ade Mahari, Florentina Bety Indah Lupitasari, Anita Hafid, Marchie Asrid da Costa, Nur Adianto, and Pradita Iustitia Sitaresmi

Volume-18 | Issue-6 | Article-20 | https://doi.org/10.14202/vetworld.2025.1614-1633

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Kisspeptin and leptin (LEP) are two essential proteins that play a central role in regulating reproductive hormones in small ruminants through the hypothalamic-pituitary-gonadal axis. These proteins influence the secretion of gonadotropin-releasing hormone, which, in turn, controls key hormones such as follicle-stimulating hormone and luteinizing hormone. Acting in synergy, kisspeptin and LEP also interact with other metabolic and reproductive signals, including insulin, estrogen, and neuropeptides, to coordinate reproductive function. Despite their importance, the detailed mechanisms by which these proteins operate, especially in relation to body condition score are not yet fully understood. This review explores their biological roles, interactions, and potential as markers for selecting high-performing livestock. External factors such as diet, stress, and seasonal changes can further influence their expression and activity. Understanding these pathways can support improved fertility management and the development of genetic or therapeutic strategies to enhance reproductive efficiency in goats and sheep. 
Research Article | 19 Jun 2025
Ecotoxicological effects of heavy metal contamination on reproductive health and gamete quality in female Nile tilapia (Oreochromis niloticus) from the Brantas River Basin, Indonesia
Habib Syaiful Arif Tuska, Gretania Residiwati, Anik Martinah Hariati, Anwar Sanusi, Gatot Ciptadi, Barlah Rumhayati, Hendra Susanto, and Aulanni’am Aulanni’am

Volume-18 | Issue-6 | Article-21 | https://doi.org/10.14202/vetworld.2025.1634-1643

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Heavy metal pollution in freshwater ecosystems poses a serious threat to aquatic biodiversity and food safety. This study assessed the bioaccumulation of lead, cadmium (Cd), iron (Fe), and copper, as well as their effects on histopathological alterations in vital organs and gamete quality, in female Nile tilapia (Oreochromis niloticus) from the Brantas River Basin, East Java, Indonesia. Water and fish samples were collected from five sites with varying levels of industrial and residential activity. Heavy metal concentrations were analyzed through atomic absorption spectrophotometry. Histopathological evaluations were performed on gills, liver, and ovaries, and gamete quality was assessed based on oocyte diameter, germinal vesicle breakdown (GVBD), survival, and abnormality rate. Statistical analyses included a one-way analysis of variance, Kruskal-Wallis tests, and Spearman correlation. The highest concentrations of Cd and Fe were found in Kalisari, corresponding with pronounced histopathological lesions in fish, including necrosis, inflammation, and hyperplasia in vital organs. Oocytes from heavily polluted sites exhibited significantly reduced diameters, GVBD rates, and survival rates, alongside increased abnormalities. Cd and Fe levels exhibited strong positive correlations with organ damage and negative correlations with gamete quality, particularly oocyte survival (Fe: r = −0.900). Exposure to elevated levels of Cd and Fe significantly impairs the physiological and reproductive health of female Nile tilapia in the Brantas River. The observed tissue damage and reproductive disruption underscore the ecological and public health risks associated with unchecked industrial discharge. Long-term biomonitoring and targeted pollution control strategies are urgently required to safeguard aquatic life and reduce health risks under the One Health framework.
Research Article | 19 Jun 2025
In vitro and in silico investigations of Propolis-derived phytochemicals as potential inhibitors of Plasmodium falciparum
Dhrubo Ahmed Khan, Md. Nazmul Hasan, Rachasak Boonhok, Suthinee Sungkanu, Yutatirat Singhaboot, Afsana Amin Shorna, Anamul Hasan, Kesinee Chotivanich, Polrat Wilairatana, Abolghasem Siyadatpanah, Roghayeh Norouzi, Imran Sama-ae, Watcharapong Mitsuwan, Alok K. Paul, Maria de Lourdes Pereira, Shanmuga S. Sundar, Tooba Mahboob, Christophe Wiart, Ryan V. Labana, Siriphorn Chimplee, and Veeranoot Nissapatorn

Volume-18 | Issue-6 | Article-22 | https://doi.org/10.14202/vetworld.2025.1644-1659

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Malaria continues to pose a global health challenge, exacerbated by the emergence of drug-resistant strains of Plasmodium falciparum. This study aimed to evaluate the anti-Plasmodium potential of Propolis extracts collected from various Iranian regions and to characterize the molecular interactions of their bioactive phytochemicals with P. falciparum lactate dehydrogenase (PfLDH), a key enzyme in parasite glycolysis. The anti-Plasmodium activity of ethanol-extracted Propolis was assessed against P. falciparum NF54 using the SYBR Green I fluorescence assay. Gas chromatography-mass spectrometry (GC-MS) analysis identified major phytochemicals in the most active extract. Molecular docking and 100-ns molecular dynamic (MD) simulations were performed to evaluate the binding affinity and stability of selected compounds (tectochrysin and galangin) against PfLDH in both holo (Protein Data Bank [PDB] ID: 1LDG) and apo (PDB ID: 2X8L) forms. Propolis collected from Kermanshah city exhibited the highest anti-Plasmodium activity (IC50 = 6.69 ± 1.44 μg/mL). GC-MS analysis identified tectochrysin and galangin as major constituents. Molecular docking revealed strong binding affinities of tectochrysin (−7.8 kcal/mol) and galangin (−7.5 kcal/mol) to PfLDH, surpassing the binding energies of standard antimalarial drugs (chloroquine and quinine). MD simulations confirmed the stability of tectochrysin and galangin within the PfLDH active sites, with favorable root mean square deviation, root mean square fluctuation, gyration, solvent-accessible surface area, molecular surface area, and polar surface area profiles, indicating persistent and stable protein-ligand interactions throughout the simulation. The findings support the promising anti-Plasmodium potential of Propolis-derived compounds, particularly tectochrysin and galangin, as novel PfLDH inhibitors. Their potential applicability in transdisciplinary anti-parasitic therapy across human and veterinary medicine warrants further in vivo validation and clinical investigations.
Research Article | 19 Jun 2025
Viability of Trichinella spiralis in traditional sour pork fermentation and its inactivation by microwave heating: Implications for zoonotic risk and food safety
Atchara Artchayasawat, Benjamabhorn Pumhirunroj, Sukhonthip Khueangchiangkhwang, Thidarut Boonmars, Parichart Boueroy, Porntip Laummaunwai, and Panaratana Rattanasuwan

Volume-18 | Issue-6 | Article-23 | https://doi.org/10.14202/vetworld.2025.1660-1666

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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.
Research Article | 19 Jun 2025
Controlled freezing inactivates Trichinella britovi in wild boar meat: Insights from a murine infection model and multiplex polymerase chain reaction
Olimpia C. Iacob, Aurelian-Sorin Pașca, Laura Andree Olariu, Larisa Maria Ivănescu, and Mihai Mareș

Volume-18 | Issue-6 | Article-24 | https://doi.org/10.14202/vetworld.2025.1667-1674

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Trichinellosis remains a public health concern globally due to the zoonotic potential of consuming undercooked meat infected with Trichinella spp. larvae. Trichinella britovi, known for its moderate freeze tolerance, presents a food safety challenge, particularly in game meat such as wild boar. This study aimed to evaluate the infectivity of T. britovi larvae in wild boar meat subjected to prolonged freezing under controlled conditions. Muscle samples (50 g each) from a wild boar naturally infected with T. britovi were frozen for 56 days at four temperatures: −18°C, −20°C, −29°C, and −40°C. Post-thaw, larval viability was assessed through artificial digestion, and infectivity was tested in a murine model using BALB/c mice. Each experimental group (n = 5 mice) received 120 larvae through gavage over 2 days. After 56 days, mice were euthanized, and muscle tissues were examined histologically. Molecular confirmation was performed using multiplex polymerase chain reaction on formalin-fixed tissues. Despite larval motility post-thaw, no viable T. britovi DNA was detected in the muscle tissues of infected mice. Histological examination showed structures resembling Trichinella cysts in all experimental groups, but these were not molecularly confirmed. The control group remained negative throughout. Controlled freezing at temperatures as low as −18°C for 8 weeks rendered T. britovi larvae in wild boar meat non-infectious in a murine model. These findings suggest that freezing may be a viable strategy for reducing the risk of trichinellosis transmission through game meat. However, given species-specific variability and environmental influences, further studies across diverse conditions are warranted to refine food safety protocols.

Volume-18 | Issue-6 | Article-25 | https://doi.org/10.14202/vetworld.2025.1675-1684

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Mycoplasma bovis is a significant pathogen in cattle, causing respiratory, reproductive, and mammary diseases, leading to substantial economic losses. Conventional control measures remain ineffective due to antimicrobial resistance and the absence of an approved vaccine. This study aimed to develop a multiepitope messenger RNA (mRNA)-based vaccine against M. bovis using immunoinformatic and molecular modeling approaches. Two conserved surface-exposed proteins – lipoate protein ligase (LplA) and dihydrolipoamide dehydrogenase (PdhD) – were selected as vaccine targets. T- and B-cell epitopes were predicted using Immune Epitope Database and evaluated for antigenicity, allergenicity, toxicity, and conservancy. Selected epitopes were linked using specific amino acid linkers and combined with a resuscitation-promoting factor E (RpfE) adjuvant and untranslated regions (hemoglobin subunit beta and rabbit beta-globin) to improve translation and stability. The vaccine construct was modeled and validated through physicochemical profiling, secondary and tertiary structure prediction, molecular-docking with bovine toll-like receptors 4 (TLR4), and codon optimization. Molecular dynamics simulations were conducted to assess the stability of the vaccine-receptor complex. The modeled vaccine construct contained five cytotoxic T lymphocyte, six helper T lymphocyte, and five B-cell epitopes. The construct was predicted to be highly antigenic (score: 0.835), non-allergenic, and non-toxic. Structural validation showed 93.5% of residues in favored regions of the Ramachandran plot and a Z-score of −10.6. Docking simulations revealed strong binding affinity to bovine TLR4, supported by robust molecular dynamics simulation outcomes, including high stability, low eigenvalues, and favorable covariance patterns. Codon optimization yielded a guanine-cytosine content of 59.8% and a codon adaptation index of 0.87, indicating efficient expression in cattle. The predicted mRNA structure exhibited good thermodynamic stability (minimum free energy: −321.42 kcal/mol). This study presents a computationally designed mRNA vaccine candidate against M. bovis based on LplA and PdhD epitopes. The construct demonstrated promising immunogenicity, structural integrity, and receptor-binding properties, representing a viable vaccine strategy. Nonetheless, in vitro and in vivo validation is essential to confirm the construct’s efficacy and safety in cattle.