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Volume 18 | May
Research Article | 08 May 2025
One Health serosurveillance of anti-SARS-CoV-2 antibodies in domestic animals from the metropolitan area of Panama
Sulamith Del C. Pacheco, Alanis J. Jimenez, Giselle A. Rangel, and Claudia Del C. Rengifo-Herrera

Volume-18 | Issue-5 | Article-1 | https://doi.org/10.14202/vetworld.2025.1082-1089

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The COVID-19 pandemic, caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), has raised concern regarding anthropozoonotic transmission to domestic animals, posing potential public and veterinary health risks. Latin America remains underrepresented in seroepidemiological assessments of such zoonotic spillover. This study aimed to detect anti-SARS-CoV-2 antibodies in domestic dogs and cats in Panama using a One Health surveillance framework. A cross-sectional serological survey was conducted between October 2022 and December 2023 across the metropolitan area of Panama City. Serum samples from 341 animals (198 dogs and 143 cats) were analyzed using a commercial double-antigen enzyme-linked immunosorbent assay to detect antibodies targeting the SARS-CoV-2 nucleocapsid protein. A historical panel of 100 pre-pandemic canine and feline samples was also tested. Demographic, clinical, and exposure data were collected through owner questionnaires, and statistical associations with seropositivity were assessed using univariate tests and binary logistic regression. Seropositivity was detected in 12/341 animals (3.5%; 95% confidence interval: 1.96–6.11%), comprising 9 dogs (4.5%) and 3 cats (2.1%). In addition, 2/100 pre-pandemic canine samples (2.0%) tested positive. Most seropositive animals (75%) were reported to have lived in households with confirmed COVID-19 cases, although this variable was not statistically associated with seropositivity. Regression analysis identified ideal body condition as a significant predictor (p = 0.016), while sampling location and demographic variables were not significant. This study presents the first serological evidence of SARS-CoV-2 exposure in domestic pets in Panama. While low in prevalence, the findings underscore the relevance of community-based animal surveillance and reveal possible serological cross-reactivity with endemic canine coronaviruses. The data support the continued integration of domestic animal monitoring into One Health strategies to preempt zoonotic risks and improve pandemic preparedness.

Volume-18 | Issue-5 | Article-2 | https://doi.org/10.14202/vetworld.2025.1090-1100

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Ticks are critical vectors of pathogens affecting humans and livestock globally. The microbiome of ticks, comprising diverse bacterial communities, plays a crucial role in tick biology and vector competence. Hyalomma ticks are prominent in the Middle East and North Africa (MENA) region and are known carriers of significant pathogens. This study aimed to systematically evaluate existing literature regarding the microbiome composition of Hyalomma ticks in the MENA region, identify predominant bacterial genera, and highlight knowledge gaps. A systematic literature search was conducted using four databases: ScienceDirect, PubMed, Google Scholar, and Scopus. The search covered studies published between 2014 and 2024 employing 16S ribosomal RNA gene sequencing to analyze microbiomes of Hyalomma ticks within the MENA region. Studies not fulfilling these criteria were excluded through independent assessment by two authors. Out of 1,220 screened articles, seven studies met inclusion criteria, involving five Hyalomma species: Hyalomma dromedarii, Hyalomma anatolicum, Hyalomma excavatum, Hyalomma marginatum, and Hyalomma scupense. Most studies (57.14%) focused on H. dromedarii, primarily collected from camels. The geographical distribution of studies included the United Arab Emirates (42.86%), Saudi Arabia (28.57%), Iran (14.29%), and Tunisia (14.29%). Common bacterial genera identified across multiple studies included Acinetobacter, Bacillus, Flavobacterium, Francisella, Rickettsia, Staphylococcus, Pseudomonas, and Corynebacterium. However, substantial gaps were noted, particularly concerning variations related to tick lifecycle stages, host interactions, temporal dynamics, and extensive geographic coverage within the MENA region. This systematic review underscores the presence of key bacterial genera within Hyalomma ticks across the MENA region, revealing their potential roles in tick biology and pathogen transmission. Major research gaps identified include limited geographical scope, insufficient exploration of microbiome variation across tick life stages, host-specific interactions, and the environmental factors influencing microbial communities. Addressing these gaps through comprehensive, longitudinal, and multi-regional studies is imperative for improving public health strategies and developing targeted tick-control methods.  
Research Article | 12 May 2025
A lung cancer mouse model system based on an inbred C3H strain: Ultrasound imaging, pathological analysis, and proteomic biomarker identification
Ulayatul Kustiati, Wahyu Tri Widayati, Dwi Liliek Kusindarta, Dwi Aris Agung Nugrahaningsih, Dinda Aliffia, Wilda Bunga Tina Sanjaya, and Hevi Wihadmadyatami

Volume-18 | Issue-5 | Article-3 | https://doi.org/10.14202/vetworld.2025.1101-1108

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Lung cancer remains a leading cause of global mortality, necessitating robust animal models for research into its mechanisms and therapeutic options. This study aimed to develop and validate a novel lung cancer mouse model using the inbred C3H strain through intraperitoneal (I.P) injection of benzo(a)pyrene, offering insights into hematology, pathology, imaging, and proteomic biomarkers. Twelve male inbred C3H mice were assigned to non-treated and treatment groups, with the latter receiving 100 mg/kg body weight of benzo(a)pyrene intraperitoneally. Tumor development was monitored for 15 days using hematological analysis, ultrasound imaging (Vevo F2), histopathological assessment, and proteomic profiling through liquid chromatography-high-resolution mass spectrometry. Hematological analysis indicated a decrease in white blood cells, lymphopenia, and neutropenia, while red blood cells, hemoglobin, and platelets remained within normal ranges. Ultrasound imaging revealed tumor formation as hypoechoic areas with irregular patterns on the lung surface. The histological analysis highlighted lymphocyte infiltration, alveolar wall thickening, fibroelastosis, and dysplastic changes in the bronchial epithelium. Proteomic profiling identified specific biomarkers associated with lung cancer, including A disintegrin and metalloproteinase with thrombospondin motifs 12, abnormal spindle, adducin-3, adhesion G protein-coupled receptor, Agrin, apoptotic chromatin condensation inducer 1, rapidly accelerated fibrosarcoma isoforms B oncogene, breast cancer gene 2, hypoxia-induced gene-1, leucine-rich repeat-containing 2, leucine-rich repeat kinase 2, leucine-rich repeat-containing protein 2 isoform X2, membrane-spanning 4-domains, subfamily A, proto-oncogene tyrosine-protein kinase Src, rat sarcoma virus-related protein 14, squamous cell carcinoma antigen, and transcription intermediary factor 1-alpha. The I.P administration of benzo(a)pyrene in C3H mice effectively induced lung cancer, demonstrating significant pathological and biomarker changes. This model provides a valuable platform for investigating lung cancer mechanisms, evaluating new therapeutic approaches, and potentially shortening the timeframe required to establish reliable animal models for preclinical studies.
Research Article | 13 May 2025
Impacts of early postpartum behavioral patterns on the fertility and milk production of tropical dairy cows
Aqeel Raza, Kumail Abbas, Theerawat Swangchan-Uthai, Henk Hogeveen, and Chaidate Inchaisri

Volume-18 | Issue-5 | Article-4 | https://doi.org/10.14202/vetworld.2025.1109-1126

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Early postpartum behavioral patterns are pivotal indicators of dairy cow health, reproductive success, and lactation performance, particularly under the environmental stressors of tropical climates. This study aimed to investigate how these behavioral patterns, as captured by smart biosensor data, influence reproductive outcomes, and milk yield in Holstein Friesian cows, with specific emphasis on parity differences and behavioral clustering. A total of 227 Holstein Friesian cows, categorized by parity (primiparous vs. multiparous), were monitored using AfiTag-II accelerometers from 3 days prepartum to 30 days postpartum. Behavioral variables – activity, rest time, rest per bout, and restlessness ratio – were subjected to K-means clustering to identify distinct behavioral profiles. Reproductive performance was analyzed using Cox proportional hazard models, while lactation dynamics were modeled using the Wood function to estimate peak yield, peak time, and persistency. Three distinct behavioral clusters were identified. Primiparous cows in Cluster 1 showed the highest early postpartum activity (~300 min/day at 5 days in milk [DIM]) and restlessness ratios, while multiparous cows exhibited more stable behavioral profiles. Cox regression suggested that cows in Cluster 0 had a higher, although non-significant, likelihood of estrus onset at 40 DIM (Hazard ratio = 1.44, p = 0.09). Lactation modeling revealed that multiparous cows in Cluster 0 attained the highest cumulative milk yield (4896.6 ± 252.1 kg at 305 DIM), while the single cow in Cluster 2 exhibited an atypical lactation curve with a delayed peak and reduced persistency. Postpartum behavioral clustering reveals parity-specific lactation and reproductive trajectories in tropical dairy cows. Higher activity and restlessness ratios may delay estrus and compromise milk yield, underscoring the potential of behavioral monitoring for targeted reproductive and nutritional management. Integration of sensor-based clustering with routine herd monitoring may support early identification of cows at risk of suboptimal performance, improving reproductive efficiency and milk production in tropical dairy systems.
Research Article | 13 May 2025
Competing microorganisms with exclusion effects against multidrug-resistant Salmonella Infantis in chicken litter supplemented with growth-promoting antimicrobials
María Alejandra Ospina Barrero, Maryeimy Varón-López, and Lina M. Peñuela-Sierra

Volume-18 | Issue-5 | Article-5 | https://doi.org/10.14202/vetworld.2025.1127-1136

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The widespread use of antibiotic growth promoters (AGPs) in poultry production has been implicated in altering gut microbiota and promoting the excretion of multidrug-resistant (MDR) bacteria into the environment. Salmonella enterica serovar Infantis (Salmonella Infantis [S.I]), a prevalent zoonotic pathogen, has demonstrated increasing resistance in poultry systems. This study aimed to evaluate the efficacy of natural control microorganisms (NCM), Bacillus subtilis and Lactobacillus plantarum, in reducing the abundance of MDR S.I in fresh chicken litter from birds raised with or without AGP supplementation. It also examined how physicochemical properties and microbial dynamics influence pathogen persistence. Microcosms were constructed using litter from broilers raised under two dietary regimes (with and without avilamycin). Treatments included combinations of AGP, S.I, and NCM. Bacterial enumeration was performed using selective media, and whole-genome sequencing of S.I was conducted to characterize antimicrobial resistance and virulence genes. Physicochemical parameters (pH, humidity, temperature, and ammonia) were measured and correlated with microbial loads. Antagonistic activity of NCM strains was assessed using agar diffusion assays. Genome analysis revealed that S.I carried multiple resistance genes (e.g., blaCTX-M-65, tet(A), and sul1) and efflux systems conferring MDR. In vitro assays showed strong antagonism by L. plantarum and moderate activity by B. subtilis. In microcosms, S.I counts significantly decreased in the presence of both AGP and NCM, indicating synergistic inhibition. Conversely, in the absence of AGP, NCM had a limited effect. Statistical analyses showed strong correlations between microbial groups and physicochemical variables, particularly during later production stages. The application of B. subtilis and L. plantarum in chicken litter significantly reduced S.I colonization under AGP supplementation, suggesting their potential as biocontrol agents. These findings support the development of integrated litter management strategies to mitigate zoonotic and resistant pathogen dissemination, particularly in AGP-using systems. However, the effectiveness of such interventions may vary across farms due to differences in microbial ecology and environmental conditions.
Research Article | 13 May 2025
Protective effect of purple sweet potatoes (Ipomoea batatas L.) against rat breast cancer
Carmen R. Silva-Correa, Julio Hilario‑Vargas, Abhel A. Calderón-Peña, Víctor E. Villarreal-La Torre, Cinthya L. Aspajo-Villalaz, Natalia Bailon-Moscoso, Juan Carlos Romero-Benavides, Oscar Herrera-Calderon, William A. Sagástegui-Guarniz, Julio A. Castañeda-Carranza, Walter E. Janampa-Castillo, and José L. Cruzado-Razco

Volume-18 | Issue-5 | Article-6 | https://doi.org/10.14202/vetworld.2025.1137-1146

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Breast cancer is one of the most prevalent and lethal malignancies affecting women worldwide. Given the limitations of conventional treatments, there is an increasing interest in exploring naturally derived compounds with chemoprotective properties. Purple sweet potatoes (Ipomoea batatas L.) are rich in anthocyanins and have been reported to possess antioxidant, anti-inflammatory, and anticancer activities. This study aimed to evaluate the chemopreventive potential of ethanolic extracts from purple sweet potato peels in a rat model of 7,12-dimethylbenz(a)anthracene (DMBA)-induced breast cancer. Fifty female Rattus norvegicus (170–200 g) were randomized into five groups. Breast tumors were induced through a single subcutaneous dose of DMBA (20 mg/rat). Three experimental groups received daily oral administration of the extract at 200, 400, and 600 mg/kg body weight, respectively, for 4 months. One control group received only DMBA, while another received the highest dose of the extract without DMBA. Antioxidant activity was assessed using 2,2-diphenyl-1-picrylhydrazyl-hydrate (DPPH) and 2,2’-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays. Anthocyanin content was quantified using spectrophotometry. Tumor latency, tumor volume, and histopathological alterations were evaluated to determine the extract’s chemopreventive effects. The extract exhibited significant antioxidant activity comparable to quercetin at 1500 ppm (DPPH assay) and a high anthocyanin content (138.92 ± 0.58 mg/100 g dry extract). Tumor latency was significantly prolonged in the 600 mg/kg group (101 days) compared to the DMBA control (88 days). In addition, this group showed a marked reduction in tumor volume (2.26 cm3 vs. 15.21 cm3; p < 0.05). Histological examination revealed improved ductal epithelial integrity and reduced necrosis in extract-treated groups, particularly at the highest dose. The ethanolic extract of purple sweet potato peels demonstrated a dose-dependent chemopreventive effect against DMBA-induced breast cancer in rats. The extract’s high anthocyanin content likely contributed to its antioxidant and antitumor activities. These findings suggest potential applications in dietary chemoprevention, warranting further investigation into its molecular mechanisms and clinical translation.
Research Article | 13 May 2025
Sulfated Lewis A from the oviduct reservoir selectively binds to camel (Camelus dromedarius) sperm and extends their lifespan in vitro
Mohamed M. M. El-Sokary, Hamad A. Albreiki, Salem Belal, and Latifa R. Alshamsi

Volume-18 | Issue-5 | Article-7 | https://doi.org/10.14202/vetworld.2025.1147-1155

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Camel reproduction faces significant challenges, including poor semen preservation and a limited understanding of gamete interactions, particularly within the oviduct. Glycan-mediated sperm binding in the oviduct is pivotal for sperm storage and longevity in various species. This study aimed to evaluate the binding affinity of camel epididymal sperm to sulfated Lewis A (SuLeA) – a trisaccharide from the oviductal isthmus – and investigate its effect on sperm lifespan and viability in vitro. Fluorescent-labeled SuLeA was used to localize glycan-binding sites on camel sperm. An in vitro model involving biotinylated SuLeA conjugated to streptavidin-sepharose beads was developed to mimic oviductal interactions. Sperm-oviduct binding specificity was assessed by pre-incubating sperm with SuLeA before their exposure to epithelial cell aggregates. Sperm viability was evaluated over 48 h using SYBR-14 and propidium iodide staining. Fluorescent SuLeA showed preferential binding to the post-acrosomal region of camel sperm (53%, p < 0.05). Pre-incubation with SuLeA significantly inhibited sperm adhesion to oviductal aggregates (82% vs. 25%, p < 0.05), confirming binding specificity. Sperm demonstrated a high affinity to immobilized SuLeA (5 sperm/bead), which was reduced to 1 sperm/bead following glycan pre-incubation. Notably, sperm bound to immobilized SuLeA exhibited significantly higher viability (59%) after 48 h compared to unbound sperm (5%, p < 0.05). This study establishes that SuLeA selectively binds to camel sperm at the post-acrosomal region, mimicking physiological sperm-oviduct adhesion. The interaction not only confirms glycan specificity but also significantly prolongs sperm viability. These findings provide a promising foundation for developing freeze-free preservation techniques and improving artificial insemination protocols in camelids.
Research Article | 13 May 2025
Ovicidal effect of eucalyptus wood vinegar on gastrointestinal nematodes’ eggs from sheep
Yandra Thais Rocha da Mota, Alexandre Santos Pimenta, Moacir Franco de Oliveira, Ana Karolinne de Alencar França, Andressa Marcelly Silvestre Pereira, Rafael Rodolfo de Melo, Thays Vieira da Costa Monteiro, Maíra Fasciotti, Lúcio César Dantas de Medeiros, and Ana Carla Diógenes Suassuna Bezerra

Volume-18 | Issue-5 | Article-8 | https://doi.org/10.14202/vetworld.2025.1156-1167

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Gastrointestinal nematodes (GINs) significantly impair small ruminant production globally, particularly in tropical regions. Anthelmintic resistance due to the indiscriminate use of synthetic drugs has necessitated the search for sustainable, plant-based alternatives. Eucalyptus wood vinegar (WV), a by-product of biomass pyrolysis, possesses bioactive compounds with potential anthelmintic activity. This study aimed to assess the in vitro ovicidal efficacy of eucalyptus WV and WV derived from co-pyrolysis of eucalyptus wood with Origanum majorana (marjoram) against eggs of GINs from naturally infected sheep. WV samples were produced through controlled pyrolysis and refined through sequential vacuum distillation. Egg hatchability tests were performed using five WV concentrations (0.3125%–5% g/100 mL), with thiabendazole as a positive control and distilled water as a negative control. Egg counts, species identification, and scanning electron microscopy (SEM) were conducted to evaluate structural changes. The chemical compositions of the WVs were characterized using gas chromatography-mass spectrometry (GC/MS). Both WVs exhibited significant ovicidal activity, with eucalyptus WV achieving 97% inhibition at 1.25%, and the marjoram-enriched WV reaching 100% inhibition at 5%. GC/MS analysis revealed the presence of phenolic compounds, furfural, thymol, and eucalyptol, the latter two being exclusive to the marjoram formulation. SEM micrographs confirmed morphological deformations in treated eggs, including loss of symmetry and membrane integrity. The synergistic interaction among bioactive components, particularly thymol, eucalyptol, and furfural, is proposed as the mechanism enhancing ovicidal activity. Eucalyptus WV, particularly when enriched with O. majorana through co-pyrolysis, exhibits potent ovicidal effects against GINs in sheep. These findings support the potential use of WVs as eco-friendly anthelmintic alternatives in integrated parasite management strategies for small ruminants.
Research Article | 17 May 2025
Radiation-induced skin regeneration: A comparative efficacy and safety analysis of alpha, beta, and gamma modalities in murine models
Madyan Ahmed Khalaf, Marwan Noori Mohammed, Baida M. Ahmed, and Sahar A. H. Al-Sharqi

Volume-18 | Issue-5 | Article-9 | https://doi.org/10.14202/vetworld.2025.1168-1179

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The therapeutic application of ionizing radiation in wound healing, especially with alpha, beta, and gamma modalities, remains largely unexplored despite its potential for enhancing regenerative processes. This study aimed to comparatively analyze the efficacy and safety of alpha radiation (IG-A), beta radiation (IG-B), and gamma radiation (IG-G) modalities in promoting skin regeneration using a murine model of full-thickness excisional wounds. Twenty male BALB/c mice were randomized into four groups (n = 5 per group): IG-A, IG-B, IG-G, and an untreated control group (CG). Following surgical induction of full-thickness wounds (8 mm diameter), irradiation groups received 15 min of exposure at four intervals post-surgery using americium-241 (alpha), strontium-91 (beta), and cesium-137 (gamma). Wound healing was monitored macroscopically and microscopically on days 0, 2, 4, 6, 8, and 10. Histological and biochemical assessments included collagen synthesis, epithelialization, neovascularization, and growth factor (vascular endothelial growth factor [VEGF] and platelet-derived growth factor [PDGF]) quantification. Statistical analysis was performed using a one-way analysis of variance. IG-A significantly accelerated wound healing, achieving approximately 100% wound closure by day 10 compared to 90% and 80% in beta and gamma radiation groups, respectively. Control wounds demonstrated only 38% closure. Histopathological analysis indicated enhanced collagen deposition, neovascularization, sebaceous gland regeneration, and complete epithelialization primarily in the alpha-treated group. Biochemical assays revealed significantly elevated VEGF and PDGF levels in irradiated groups, with IG-A exhibiting the highest expression. IG-A demonstrated superior efficacy in accelerating wound healing and tissue regeneration compared to beta and gamma modalities. This novel finding suggests a potential therapeutic role for IG-A in clinical wound management strategies.
Research Article | 17 May 2025
Potency of Lacticaseibacillus paracasei as an alternative to antibiotic growth promoter in broiler chicken challenged with avian pathogenic Escherichia coli
Widya Paramita Lokapirnasari, Lilik Maslachah, Amung Logam Saputro, Efi Rokana, Andreas Berny Yulianto, Zulfi Nur Amrina Rosyada, Muhammad Aviv Firdaus, Himatul Ilma Silfia, Ertika Fitri Lisnanti, Zein Ahmad Baihaqi, Tabita Dameria Marbun, Aswin Rafif Khairullah, and Muhammad Shakeel

Volume-18 | Issue-5 | Article-10 | https://doi.org/10.14202/vetworld.2025.1180-1189

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The emergence of antibiotic-resistant bacteria due to the widespread use of antibiotic growth promoters (AGPs) necessitates the exploration of sustainable alternatives in poultry production. This study evaluated the efficacy of Lacticaseibacillus paracasei as a probiotic alternative to AGPs in broilers challenged with avian pathogenic Escherichia coli (APEC), with a focus on growth performance, antioxidant status, lipid metabolism, and hematological profiles. A total of 80 broiler chickens were randomly allocated into four groups: T0 (uninfected control), T1 (APEC-infected control), T2 (APEC + AGP, 0.1% zinc bacitracin), and T3 (APEC + probiotic, 0.5% L. paracasei). Treatments were administered from day 15 to 35 post-hatch, following a 2-week adaptation period. Performance indicators (feed conversion ratio [FCR], body weight gain [BWG], and feed efficiency [FE]), antioxidant parameters (superoxide dismutase [SOD] and malondialdehyde [MDA]), lipid profiles (cholesterol, HDL, and LDL), and hematological variables were assessed. Data were analyzed using analysis of variance with significance set at p < 0.05. Probiotic supplementation significantly improved FCR, BWG, and FE (p < 0.05), with the T3 group achieving the most favorable outcomes. SOD activity was markedly higher, and MDA levels were reduced in probiotic-treated birds compared with the AGP and infected groups. Moreover, L. paracasei administration resulted in significant reductions in total cholesterol and LDL levels while maintaining moderate HDL concentrations. Hemoglobin and thrombocyte levels were modestly influenced, though overall hematological profiles remained stable across groups. L. paracasei supplementation significantly enhanced growth performance, antioxidant defense, and lipid metabolism in broilers challenged with APEC, supporting its application as a viable alternative to AGPs. These findings contribute to sustainable poultry production practices and public health safety by mitigating the reliance on antibiotics.
Review Article | 17 May 2025
Benefits of aromatase blockers for increased testosterone in poultry: A mini-review
Rizki Fitrawan Yuneldi, Claude Mona Airin, Sarmin Sarmin, Andhika Yudha Prawira, and Pudji Astuti

Volume-18 | Issue-5 | Article-11 | https://doi.org/10.14202/vetworld.2025.1190-1198

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Testosterone is a key androgenic hormone in male poultry, regulating growth performance, reproductive function, and the development of secondary sexual characteristics. However, endogenous testosterone levels are often diminished through conversion to estrogen through the aromatase enzyme, presenting a physiological constraint in poultry production systems. While synthetic testosterone administration has been employed to overcome this limitation, it is frequently accompanied by adverse effects, including gonadal atrophy and impaired spermatogenesis. Consequently, aromatase blockers have emerged as a promising strategy to enhance testosterone levels by inhibiting estrogen biosynthesis. This review synthesizes current evidence on both synthetic aromatase blockers (SABs), such as letrozole and tamoxifen, and natural aromatase blockers (NAB), including compounds derived from Anadara granosa and Anadara nodifera clamshells, plant extracts, and trace minerals like zinc. The mechanisms, efficacy, physiological effects, and safety profiles of NAB are comparatively examined against SAB. The findings indicate that NAB can significantly elevate testosterone levels and improve reproductive and performance traits without the adverse histopathological effects observed with prolonged SAB or synthetic androgen use. This review highlights the potential of NAB as sustainable alternatives to synthetic hormones in poultry production and recommends further investigations to optimize dosing regimens, elucidate long-term effects, and explore combinatorial strategies.
Review Article | 17 May 2025
Ecological and public health dimensions of ESBL-producing Escherichia coli in bats: A One Health perspective
Alfiana Laili Dwi Agustin, Aswin Rafif Khairullah, Mustofa Helmi Effendi, Wiwiek Tyasningsih, Ikechukwu Benjamin Moses, Budiastuti Budiastuti, Hani Plumeriastuti, Sheila Marty Yanestria, Katty Hendriana Priscilia Riwu, Fidi Nur Aini Eka Puji Dameanti, Wasito Wasito, Riza Zainuddin Ahmad, Agus Widodo, and Daniah Ashri Afnani

Volume-18 | Issue-5 | Article-12 | https://doi.org/10.14202/vetworld.2025.1199-1213

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The emergence and global dissemination of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli represent a major public health concern. While antibiotic resistance in clinical and agricultural settings is well documented, the contribution of wildlife, particularly bats, to the spread of antimicrobial resistance (AMR) remains underexplored. Bats possess unique ecological traits – such as long-distance flight, longevity, and adaptability – that facilitate their role as potential reservoirs and vectors of antibiotic-resistant bacteria. This review synthesizes global findings on the occurrence, genetic characteristics, and transmission dynamics of ESBL-producing E. coli isolated from bats. Through a comprehensive literature review of studies conducted across five continents, we highlight the prevalence of multidrug-resistant E. coli in bat populations, with resistance profiles frequently including β-lactams (bla), aminoglycosides, tetracyclines, and fluoroquinolones. Notably, key ESBL genes such as blaCTX-M, blaTEM, blaSHV, and blaOXA have been identified in isolates from bat feces (guano), raising significant concern due to potential environmental contamination and zoonotic spillover. Risk factors such as habitat encroachment, anthropogenic waste exposure, and the agricultural use of bat guano further exacerbate the risk of ESBL transmission. Moreover, genomic comparisons suggest phylogenetic overlap between ESBL-producing E. coli from bats and those found in humans and livestock. Given these findings, bats warrant greater inclusion in One Health surveillance frameworks to trace AMR gene flow and develop targeted interventions. This review underscores the need for integrated AMR monitoring in wildlife, enhanced waste management policies, and stricter biosecurity to mitigate the public health risks associated with wildlife-origin ESBL dissemination.
Research Article | 17 May 2025
A comparison of diagnostic methods for canine Ehrlichiosis: Microscopy and RNases hybridization-assisted amplification technology compared with the quantitative polymerase chain reaction
Wisut Prasitsuwan, Thanikran Suwannachote, Thirawat Sumalai, Rachakris Lertpatarakomol, Tassanee Trairatapiwan, and Sakchai Ruenphet

Volume-18 | Issue-5 | Article-13 | https://doi.org/10.14202/vetworld.2025.1214-1223

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Canine vector-borne pathogens, particularly blood parasites, pose significant health threats to domestic dogs, ranging from subclinical infections to severe systemic diseases. In Thailand, microscopic examination remains the standard diagnostic method, despite its limitations. This study aimed to (i) determine the prevalence of major canine vector-borne pathogens in Bangkok, Thailand during the 2024 rainy season and (ii) evaluate the diagnostic performance of microscopy and a novel RNase hybridization-assisted amplification (RHAM) test kit in detecting canine Ehrlichiosis, compared to the quantitative polymerase chain reaction (qPCR) gold standard. A total of 134 whole blood samples were collected from clinically suspected dogs. Microscopy was performed on Giemsa-stained blood smears, and the RHAM test kit was employed for nucleic acid detection. qPCR served as the reference method. Sensitivity, specificity, accuracy, and precision of each diagnostic method were calculated relative to qPCR results. Microscopic examination revealed the following infection prevalences: Ehrlichia spp. (26.12%), Babesia spp. (4.48%), Hepatozoon canis (6.72%), Anaplasma spp. (0.75%), Dirofilaria immitis (3.73%), and Brugia spp. (3.73%). Compared with qPCR, microscopy demonstrated a sensitivity of 51.47%, specificity of 87.88%, accuracy of 69.40%, and precision of 81.39% for Ehrlichiosis detection. In contrast, the RHAM test kit achieved markedly higher diagnostic metrics: Sensitivity (91.18%), specificity (98.48%), accuracy (94.78%), and precision (98.41%). Notably, the RHAM kit provided rapid, user-friendly detection, approximating qPCR diagnostic performance, although its sensitivity slightly declined in samples with very low pathogen titers. This study highlights the continued high prevalence of Ehrlichiosis among dogs in Bangkok during the rainy season. Although microscopy remains practical, its diagnostic limitations are significant. The RHAM test kit demonstrated excellent sensitivity and specificity, offering a rapid and accurate alternative for Ehrlichiosis detection, particularly suitable for resource-limited settings lacking qPCR capabilities. Adoption of the RHAM assay could improve early diagnosis and management of canine Ehrlichiosis at grassroots veterinary facilities.
Research Article | 17 May 2025
Innovative use of indigenous dadih probiotics to enhance feed intake, digestibility, growth performance, and health in heat-stressed Sapera goats
Nurzainah Ginting, Edhy Mirwandhono, Nurjama’yah Br. Ketaren, and Yuan-Yu Lin

Volume-18 | Issue-5 | Article-14 | https://doi.org/10.14202/vetworld.2025.1224-1233

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Heat stress resulting from rising ambient temperatures in tropical climates poses a significant threat to ruminant productivity, leading to suppressed feed intake, impaired growth, and reduced health. Indigenous fermented foods such as dadih – a traditional probiotic made from fermented buffalo milk in bamboo tubes – may offer a sustainable nutritional intervention. This study aimed to investigate the effects of dadih supplementation on feed consumption, nutrient digestibility, growth performance, pathogenic bacterial load, and hematological profiles of heat-stressed Sapera goats. The dominant bacterial strain in dadih was characterized using 16S ribosomal RNA sequencing and evaluated for in vitro antagonism against Escherichia coli and Salmonella spp. An in vivo trial was conducted using 15 Sapera crossbred goats (15 ± 1.46 kg), randomly allocated into three treatment groups (n = 5): 8 cc dadih/day (Group A), 4 cc/day (Group B), and control (Group C). The trial lasted 4 weeks, during which feed consumption, daily weight gain, feed efficiency (FE), fecal pathogenic bacteria load, and hematological parameters were measured. The probiotic strain was identified as Lactiplantibacillus plantarum Japan collection of microorganisms 1149, exhibiting antibacterial activity with inhibition zones of 9.3 mm (E. coli) and 9.5 mm (Salmonella). Goats supplemented with 4 cc dadih (Group B) demonstrated the highest daily weight gain (127.14 g/day), FE (0.15), and nutrient digestibility. A higher dadih dose (8 cc) significantly reduced fecal E. coli levels. Hematological indices remained within normal physiological ranges across all treatments, suggesting no adverse effects. This study provides the first empirical evidence supporting the use of dadih as a climate-adaptive probiotic intervention in goats. Supplementation with 4 cc dadih optimized performance without disrupting hematological homeostasis, while 8 cc effectively suppressed gut pathogens. These findings offer novel insights into the functional role of traditional fermented probiotics in improving resilience to heat stress and promoting sustainable small ruminant production in tropical environments.
Research Article | 21 May 2025
Effects of melatonin on in vitro oocyte maturation and embryo development in pigs
Laura Andrea Blancas-Alvarez, Alma Lilia Alvarez-Guerrero, Alicia Alcantar-Rodriguez, and Alfredo Medrano

Volume-18 | Issue-5 | Article-15 | https://doi.org/10.14202/vetworld.2025.1234-1241

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In vitro fertilization (IVF) efficiency in pigs remains suboptimal, partly due to oxidative stress during oocyte maturation and embryo development. Melatonin (MLT), an endogenous antioxidant, has been proposed as a beneficial supplement in reproductive culture systems. This study aimed to evaluate the effects of different concentrations of MLT on in vitro porcine oocyte maturation and subsequent embryo development. Ovaries were obtained from prepubertal gilts at a local slaughterhouse. A total of 1142 cumulus-oocyte-complexes (COC) were allocated into four groups and matured in vitro with 0, 1, 3, or 5 μM MLT for 44 h. Oocyte maturation was assessed using aceto-orcein staining and viability with trypan blue staining. Subsequently, 1312 COC underwent IVF using a standardized sperm concentration, followed by embryo culture in North Carolina State University-23 medium supplemented with corresponding MLT concentrations for 8 days. Embryo development was classified according to cleavage, morula, and early blastocyst stages. Data were analyzed using analysis of variance with Tukey’s post hoc test (p < 0.05). Although supplementation with 1.0 μM MLT resulted in the highest metaphase II oocyte maturation rate (45.9% ± 5.70%) and blastocyst formation (26.9% ± 9.57%), no significant differences were observed among treatments in either oocyte maturation, COC viability, or embryo development stages (p > 0.05). Supplementation with 0–5 μM MLT during oocyte maturation and embryo culture did not significantly enhance in vitro maturation rates or embryonic developmental outcomes in pigs. Further investigations are warranted to optimize MLT concentrations and elucidate its mechanistic role during porcine oocyte and embryo in vitro culture.
Research Article | 21 May 2025
Virulence genes and phylogenetic analysis of antibiotic-resistant Escherichia coli isolated from pig slaughterhouses in Banten Province, Indonesia
Hadri Latif, Debby Fadhilah Pazra, Chaerul Basri, Dinda Iryawati, I. Wayan Teguh Wibawan, and Puji Rahayu

Volume-18 | Issue-5 | Article-16 | https://doi.org/10.14202/vetworld.2025.1242-1252

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Escherichia coli is a prominent zoonotic pathogen with diverse virulence factors and significant antibiotic resistance, particularly in pig farming environments. Pig slaughterhouses are critical points of potential bacterial transmission to humans and the environment. Comprehensive genomic surveillance of E. coli in these settings remains limited in Indonesia. This study aimed to investigate the phylogenetic distribution, virulence gene profiles, pathotypes, and antibiotic resistance characteristics of E. coli isolated from pig slaughterhouses in Banten Province, Indonesia, using whole-genome sequencing. Environmental samples, including effluent and floor swabs (n = 200), were collected from 10 pig slaughterhouses. E. coli isolates were identified and previously characterized for antibiotic resistance. Genomic DNA was extracted and sequenced using the Oxford Nanopore MinION platform. Bioinformatic analyses, including virulence gene detection (VirulenceFinder), phylogenetic reconstruction (RAxML), and phylogroup determination (Clermont method), were conducted to classify isolates based on pathotype and genetic lineage. Fifty-seven virulence genes were identified, including 46 associated with enteric pathotypes (Enterohemorrhagic E. coli: 35%, enterotoxigenic E. coli: 15%, and enteropathogenic E. coli: 5%) and 15 linked to extraintestinal pathotypes (uropathogenic E. coli: 95%, and neonatal meningitis E. coli: 5%). Phylogenetic analysis revealed five phylogroups - A, B1, D, G, and clade I - with A and B1 predominating. Most isolates (60%) exhibited a single pathotype, while a minority (5%) carried genes from multiple pathotypes. Serotypes O73, O78, and O157 were identified, with O73 being the most prevalent. No strong correlation was observed between phylogenetic clustering and virulence gene pathotype. The high prevalence of multidrug-resistant E. coli with diverse virulence genes in pig slaughterhouses highlights significant zoonotic and environmental health risks. These findings underscore the need for enhanced hygiene practices, antimicrobial stewardship, and longitudinal genomic surveillance in Indonesian pig production systems.

Volume-18 | Issue-5 | Article-17 | https://doi.org/10.14202/vetworld.2025.1253-1261

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The escalating global threat posed by antimicrobial resistance has intensified the search for novel antimicrobial agents. Plant-derived bioactive compounds represent a promising reservoir due to their chemical diversity and efficacy against resistant pathogens. Quercus species, traditionally utilized in herbal medicine, have shown significant bioactive potential. However, research specifically evaluating the antimicrobial and antibiofilm properties of Quercus coccifera remains limited. This study aimed to investigate the phytochemical composition, antimicrobial, antibiofilm, and antioxidant activities of Q. coccifera leaf extracts using various extraction methods and solvents with differing polarities. Q. coccifera leaves were harvested, dried, and extracted using solvents of varying polarity (n-hexane, chloroform, methanol, boiled water, and microwaved water). Phytochemical profiling included tests for alkaloids, tannins, glycosides, and terpenoids. Antioxidant activity was assessed using the DPPH assay. Antimicrobial activities against Gram-positive (Staphylococcus aureus and Streptococcus pneumoniae) and Gram-negative (Pseudomonas aeruginosa, Escherichia coli, and Brucella melitensis) bacteria were evaluated using AlamarBlue® (Invitrogen, Glasgow, UK) assay and Minimum Inhibitory Concentration (MIC) determination. Antibiofilm activity was assessed by biofilm viability tests and Minimum Biofilm Eradication Concentration (MBEC) assays. Methanolic and boiled water extracts demonstrated robust phytochemical profiles (alkaloids, tannins, glycosides, and terpenoids) and significant antioxidant activity (>90% inhibition). Antimicrobial pre-evaluation indicated superior antibacterial efficacy (>90% inhibition) of these extracts, while microwaved water extracts showed moderate activity (~75% inhibition). The methanolic and boiled water extracts exhibited potent antimicrobial effects with MIC values <30 μg/mL against all tested pathogens except S. pneumoniae. Similarly, these extracts effectively disrupted biofilms formed by S. aureus and P. aeruginosa, with MBEC values approximately 25 μg/mL. Polar solvent extracts of Q. coccifera leaves exhibit significant antimicrobial and antibiofilm activities, underlining their potential as novel antimicrobial agents or adjuncts to existing therapies. Future studies involving cytotoxicity evaluation and in vivo efficacy are essential to translate these findings into clinical applications.
Research Article | 21 May 2025
Injectable homologous platelet-rich plasma, alone or in combination with oral omega-3 supplementation, for treating keratoconjunctivitis sicca in dogs
William dos Santos Villa, João Victor Goulart Consoni Passareli, Giovana José Garcia Estanho, Marco Aurélio da Cruz Nobre Gomes, Gisele Alborghetti Nai, Cecília Laposy Santarém, and Silvia Franco Andrade

Volume-18 | Issue-5 | Article-18 | https://doi.org/10.14202/vetworld.2025.1262-1273

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Keratoconjunctivitis sicca (KCS) is a chronic inflammatory condition of the canine ocular surface primarily caused by immune-mediated destruction of lacrimal tissues. Platelet-rich plasma (PRP) is known for its regenerative and anti-inflammatory properties, while omega-3 (ω-3) fatty acids possess immunomodulatory effects. This study aimed to evaluate the efficacy of injectable homologous PRP (HPRP), alone or in combination with oral ω-3 supplementation, in improving clinical and histopathological parameters in dogs with KCS. Twenty-two dogs (44 eyes) with bilateral KCS were randomized into two treatment groups: HPRP (n = 22 eyes) and HPRP plus oral ω-3 (HPRPO; n = 22 eyes). Treatments were administered monthly for up to three sessions alongside topical lubricants. Ophthalmological evaluations – including Schirmer’s tear test-1 (STT-1), tear film breakup time (TBUT), fluorescein staining, cytology of the third eyelid gland, and conjunctival histopathology – were performed at baseline and at monthly intervals up to 6 months. Both groups demonstrated significant improvement in ocular clinical signs and tear secretion. STT-1 values significantly increased from baseline in both groups (p < 0.05), without significant intergroup differences. However, TBUT values were significantly higher in the HPRPO group from month 3 onwards (p < 0.05). Cytological and histological analyses revealed a significant reduction in lymphocyte and neutrophil counts and an increase in goblet cell numbers in both groups, with greater improvement in the HPRPO group (p < 0.05). Earlier resolution of corneal ulcers and reduced ocular inflammation were observed in the HPRPO group. Injectable HPRP, particularly when combined with oral ω-3 supplementation, is an effective therapeutic modality for managing KCS in dogs. The combination therapy enhanced tear film stability, reduced ocular inflammation, and promoted epithelial repair more effectively than HPRP alone. These findings support the synergistic effect of ω-3 fatty acids with PRP in ocular surface restoration.

Volume-18 | Issue-5 | Article-19 | https://doi.org/10.14202/vetworld.2025.1274-1287

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The aryl hydrocarbon receptor (AhR) plays a pivotal role in spermatogenesis through its regulatory functions in redox balance and gene expression. This study aimed to investigate the effects of resveratrol (RES), a polyphenolic AhR modulator, and CH223191, a selective AhR antagonist, on male reproductive function in rats by assessing sperm quality, oxidative stress, testicular histopathology, and AhR gene expression. Forty adult male rats were randomly divided into four groups: (i) Control, (ii) dimethyl sulfoxide (vehicle), (iii) RES (100 mg/kg i.p., twice weekly), and (iv) AhR⁻ (CH223191, 10 mg/kg i.p., twice weekly), treated for 60 days. Post-treatment, sperm motility, survival, viability, and DNA fragmentation were evaluated. Total antioxidant capacity (TAC), malondialdehyde (MDA) levels, testicular histopathology, and AhR gene expression quantitative polymerase chain reaction (qPCR) were analyzed. RES significantly enhanced sperm motility, survival, and viability, reduced DNA fragmentation, and increased TAC while decreasing MDA levels. Histologically, RES preserved normal testicular architecture. In contrast, AhR inhibition through CH223191 led to marked reductions in sperm quality, elevated oxidative stress, increased DNA fragmentation, and severe testicular degeneration. qPCR analysis revealed upregulation of AhR expression in the RES group (fold change: +23.1%) and significant downregulation in the AhR⁻ group (fold change: −72.6%), indicating differential modulation of AhR signaling pathways. RES positively modulates AhR activity, safeguarding testicular structure and enhancing sperm quality through antioxidant and anti-apoptotic mechanisms. Conversely, AhR antagonism disrupts spermatogenesis, underscoring the receptor’s essential role in male fertility. These findings suggest the therapeutic potential of AhR-targeting agents like RES in ameliorating male reproductive dysfunctions associated with oxidative stress and xenobiotic exposure.
Research Article | 21 May 2025
Presence of toxin-producing bacterial pathogens and associated risk factors in neonatal diarrhea of piglets on commercial sow farms in Vietnam
Duy Tien Do, Tram Thi Ngoc Ngo, Huong Dieu Vu, Nhat Minh Duong, Sittikorn Traiyarach, Danh Cong Lai, and Joaquin Miguel Escuder

Volume-18 | Issue-5 | Article-20 | https://doi.org/10.14202/vetworld.2025.1288-1296

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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. 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. 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. 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.
Research Article | 25 May 2025
Biomarker-based evaluation of aflatoxin B1 exposure in cattle
Priyadharshini Ponnusamy, Umaya Suganthi Rajendran, Madhavan Gopalakrishnan Nair, Uma Sambath, Raja Kumar, Jacob Thanislass, Avinash Warundeo Lakkawar, Vijayalakshmi Padmanaban, and Poobitha Subbarayan

Volume-18 | Issue-5 | Article-21 | https://doi.org/10.14202/vetworld.2025.1297-1305

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Assessment of aflatoxin B1 (AFB1) exposure in cattle traditionally relies on feed analysis, which may not reflect chronic exposure or accurately indicate individual susceptibility. This study aimed to evaluate the utility of serum AFB1-albumin adducts and blood AFB1-DNA adducts as biomarkers for assessing individual chronic AFB1 exposure in cattle, irrespective of immediate feed contamination levels. Blood samples were collected from 53 crossbred cattle from farms, clinical veterinary cases, and slaughterhouses in Puducherry, India. Feed samples (n = 40) from farm and clinical cases were analyzed for aflatoxin contamination using two-dimensional thin-layer chromatography. AFB1 exposure was quantified by measuring serum AFB1-albumin adducts and blood AFB1-DNA adducts using an indirect enzyme-linked immunosorbent assay. In addition, a novel method was developed to synthesize the aflatoxin B1-formamidopyrimidine (AFB1-FAPy) adduct in vitro and the synthesized adduct was characterized to serve as a standard for DNA adduct quantification. AFB1 was detected in 50% of feed samples, with 70% of positive samples exceeding the maximum permissible limit of 20 μg/kg. Despite variable feed contamination, serum AFB1-albumin and blood AFB1-DNA adducts were consistently detected across all animal groups. Median AFB1-albumin adduct levels were similar among farm (0.730 pg/mg), clinical (0.670 pg/mg), and slaughterhouse (0.770 pg/mg) cattle (p = 0.731). Median AFB1-DNA adduct levels were highest in slaughterhouse cattle (18.33 pmol/μg DNA), followed by farm (14.76 pmol/μg DNA) and clinical cases (7.47 pmol/μg DNA), although differences were not statistically significant (p = 0.328). No significant correlation was observed between feed contamination levels and biomarker concentrations, highlighting the chronic nature of AFB1 exposure. The consistent detection of AFB1-albumin and AFB1-DNA adducts in cattle, irrespective of detectable aflatoxin levels in feed, underscores the limitations of traditional feed analysis for monitoring chronic exposure. The novel synthesis and robust detection of AFB1-FAPy DNA adducts further enhance the reliability of these biomarkers. These biomarkers are minimally invasive, sensitive, and valuable for chronic aflatoxin exposure assessment, aiding proactive management strategies to safeguard animal health and public food safety.
Research Article | 25 May 2025
Polymorphism of splicing factor 3A subunit 3/PstI gene and its association with the performance of Madura cows (Bos indicus)
Widya Pintaka Bayu Putra, Hartati Hartati, Endang Tri Margawati, Mariyono Mariyono, Tulus Maulana, and Thobela Louis Tyasi

Volume-18 | Issue-5 | Article-22 | https://doi.org/10.14202/vetworld.2025.1306-1312

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Madura cattle (Bos indicus), a native Indonesian breed, are primarily raised for meat production and possess unique genetic characteristics shaped by crossbreeding with Bos javanicus, Bos taurus, and B. indicus. Despite their cultural and economic importance, limited molecular studies have explored candidate genes influencing their productive traits. This study aimed to identify the single-nucleotide polymorphism (SNP) g.1292A>T (rs473122879) in intron 2 of the Splicing Factor 3A Subunit 3 (SF3A3) gene and to evaluate its association with key phenotypic traits in Madura cows. A total of 49 adult Madura cows (>3 years) were sampled from two breeding locations: Java (n = 29) and Madura Island (n = 20). Body weight, withers height, hip height, body length (BL), and heart girth were measured using standardized morphometric techniques. Genomic DNA was extracted from whole blood and subjected to polymerase chain reaction (PCR) amplification targeting a 633 bp fragment of the SF3A3 gene. Genotyping of SNP g.1292A>T was performed using PCR-restriction fragment length polymorphism with PstI enzyme digestion. Sequencing analysis was conducted for genotype confirmation. Genetic diversity indices and Hardy–Weinberg equilibrium were assessed, and association between genotype and phenotypic traits was evaluated using a general linear model. Two genotypes, TT (0.69) and AT (0.31), were identified; the AA genotype was absent in all samples. The polymorphism was under Hardy–Weinberg equilibrium (χ2 < 3.84) with a moderate polymorphic information content (PIC = 0.23). A significant association was found between SF3A3/PstI polymorphism and BL in cows from Madura Island (p < 0.05), with heterozygous (AT) individuals exhibiting superior morphometric traits compared to TT homozygotes. The SF3A3/PstI gene is polymorphic in Madura cows and exhibits moderate genetic diversity. The presence of the g.1292A>T SNP is significantly associated with BL, particularly in animals from their native breeding environment. These findings suggest the potential utility of SF3A3/PstI as a genetic marker in molecular selection strategies aimed at improving Madura cattle productivity.
Research Article | 25 May 2025
A comparative study of collagen morphology and joint strength in anterior cruciate ligament repair and reconstruction models in rabbits
Andre Yanuar, Andi Isra Mahyuddin, Nucki Nursjamsi Hidajat, Hasrayati Agustina, Nicolaas Cyrillus Budhiparama, and Nur Atik

Volume-18 | Issue-5 | Article-23 | https://doi.org/10.14202/vetworld.2025.1313-1321

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Anterior cruciate ligament (ACL) repair offers several theoretical advantages over reconstruction, including preservation of native proprioception and reduced donor-site morbidity. However, the current experimental models are predominantly limited to ACL reconstruction, leaving a critical gap in ACL repair research. This study introduces a novel rabbit model to evaluate osteointegration and mechanical strength at the tendon/ligament-bone interface following ACL repair and reconstruction. Six male New Zealand White rabbits (Oryctolagus cuniculus), aged 90 ± 0 days and weighing 2.50 ± 0.20 kg, were randomly assigned to two groups: ACL reconstruction (n = 3) using the extensor digitorum longus tendon graft and ACL repair (n = 3) using the Krackow suture technique at the femoral attachment. Specimens were collected 6 weeks postoperatively for histological evaluation of Sharpey’s-like fibers, immunohistochemical analysis of types I and III collagen, and biomechanical tensile testing. All surgical procedures were completed without complications. Histological analysis showed greater numbers of Sharpey’s-like fibers in the reconstruction group (6.33 ± 0.58%) compared to the repair group (5.67 ± 1.6%), though not statistically significant (p > 0.05). Type I collagen fibers were significantly longer in the reconstruction group in both longitudinal (3.10 ± 0.05 μm vs. 2.97 ± 0.04 μm) and transverse (1.94 ± 0.09 μm vs. 1.81 ± 0.05 μm) dimensions (p < 0.05). Type III collagen dimensions did not differ significantly. The mean tensile failure load was higher in the reconstruction group (105.96 ± 63.37 N) than in the repair group (62.56 ± 20.11 N), though this difference was not statistically significant (p > 0.05). This study establishes a reproducible and cost-effective ACL repair model in rabbits and confirms that tendon-bone osteointegration occurs in both ACL repair and reconstruction. Superior biomechanical strength and enhanced type I collagen integration in the reconstruction group underscore current clinical outcomes favoring reconstruction. This model offers a valuable platform for exploring biological augmentation strategies to enhance ACL repair efficacy.
Research Article | 25 May 2025
Assessment of the physical and chemical properties of vaginal discharge for the diagnosis of endometritis in dairy cattle
Nest Dale Bartolome, Ruchikon Jongsuwanwattana, Sariya Asawakarn, Siriwat Suadsong, Kiattisak Sangpradit, and Theerawat Swangchan-Uthai

Volume-18 | Issue-5 | Article-24 | https://doi.org/10.14202/vetworld.2025.1322-1332

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Postpartum endometritis is a major cause of infertility in dairy cattle, impacting herd productivity and economic sustainability. Although traditional diagnostic methods are available, there remains a need for simple, rapid, and accurate cow-side diagnostic tools to facilitate early detection and management of clinical endometritis (CE) and subclinical endometritis (SCE). This study aimed to (i) evaluate the diagnostic performance of vaginal discharge parameters - pH, total dissolved solids (TDS), salinity, and electrical conductivity (EC) - using a portable flat-surface electrode probe, and pH and leukocyte esterase (LE) activity using reagent strips and (ii) establish optimal cut-off values for these parameters to support practical on-farm screening of endometritis. Fifty-eight postpartum Holstein Friesian cows were enrolled. Vaginal discharge was collected using a Metricheck device and analyzed for pH, TDS, salinity, and EC. Concurrently, pH and LE activity were assessed using reagent strips. Endometrial cytology was performed to diagnose CE and SCE. Diagnostic performance was evaluated using receiver operating characteristic (ROC) curve analysis. pH values measured by both the meter and strip were significantly higher in cows with normal uterine health compared to those with CE (p = 0.010 and p = 0.008, respectively). LE activity was significantly elevated in cows with CE (p = 0.001). ROC analysis identified optimal cut-off values: pH meter ≤8.35 (area under the curve [AUC] = 0.768) and LE strip ≥2 (AUC = 0.835) for diagnosing CE. Combining pH and LE strip results improved diagnostic performance (AUC = 0.801), achieving 65.22% sensitivity, 87.5% specificity, and 76.6% accuracy. TDS, salinity, and EC were not significantly associated with uterine health status (p > 0.05). The combined evaluation of vaginal discharge pH and LE activity offers a practical, cost-effective cow-side screening method for diagnosing endometritis in dairy cattle. In contrast, TDS, salinity, and EC measurements were not diagnostically informative. The proposed approach may enhance herd health management by enabling timely identification and treatment of endometritis.
Systematic Review | 25 May 2025
Prevalence of trypanosomiasis in domesticated animals in Indonesia: A systematic review and meta-analysis
Lintang Winantya Firdausy, Faisal Fikri, Arya Pradana Wicaksono, Hakan Çalışkan, and Muhammad Thohawi Elziyad Purnama

Volume-18 | Issue-5 | Article-25 | https://doi.org/10.14202/vetworld.2025.1333-1344

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Trypanosomiasis is a vector-borne parasitic disease with significant implications for animal health and rural livelihoods in Indonesia. Despite surveillance efforts, comprehensive national-level estimates of its prevalence in domesticated animals remain lacking. This study aimed to synthesize the pooled prevalence of trypanosomiasis across Indonesian provinces, identify contributing factors, and assess trends over time using a systematic review and meta-analysis. A systematic search was conducted in seven electronic databases (PubMed, Scopus, Web of Science, ScienceDirect, Cochrane Library, ProQuest, and Google Scholar) for articles published between 1988 and 2024. Eligible studies reported primary prevalence data of trypanosomiasis in domesticated animals within Indonesia. A total of 18 studies with 4,295 samples met the inclusion criteria. Random-effects meta-analysis was performed using R Studio 4.4.2. Subgroup analyses were conducted based on animal host, diagnostic method, province, and study period. Heterogeneity was assessed through I2 and τ2 statistics, and publication bias was evaluated using Egger’s test and funnel plots. The pooled prevalence of trypanosomiasis in domesticated animals across Indonesia was 31.23% (95% confidence interval: 24.67–37.78), with considerable heterogeneity (I2 = 98.1%). Buffaloes exhibited the highest infection rate at 51.46%, followed by cattle (33.99%), whereas horses and dogs had notably lower rates (<6%). Provinces with the highest reported prevalence included Lampung (75.05%) and Central Kalimantan (75.00%). Enzyme-linked immunosorbent assay was the most frequently used and sensitive diagnostic method. Meta-regression revealed a declining trend over time (p = 0.0002), although high variability persisted between regions and diagnostic tools. Trypanosomiasis remains endemic among domesticated animals in Indonesia, with a substantial pooled prevalence and marked regional variability. The findings underscore the need for improved surveillance, implementation of standardized diagnostic tools, and integrated vector management strategies. Future research should focus on ecological risk factors, seasonality, and the zoonotic potential of Trypanosoma evansi to support evidence-based control interventions.
Research Article | 25 May 2025
Retinal vascular calibers and correlations with biomarkers in bitches with pyometra-induced systemic inflammatory response syndrome
Fábio Dumit Pizzinatto, Alexandre Pinto Ribeiro, Bianca Eidt Rodrigues, Hanna Rodrigues Miranda, and Nathalia de Assis Pereira

Volume-18 | Issue-5 | Article-26 | https://doi.org/10.14202/vetworld.2025.1345-1356

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Pyometra is a common uterine disease in intact bitches, frequently progressing to systemic inflammatory response syndrome (SIRS). While retinal vascular alterations have been observed in human SIRS cases, similar evaluations are lacking in veterinary medicine. This study aimed to evaluate retinal vascular calibers in bitches with pyometra-induced SIRS (P-SIRS) and explore correlations between retinal measurements and hematological, biochemical, vascular endothelial growth factor (VEGF), and interleukin-4 (IL-4) levels. A prospective observational study was conducted on 40 bitches diagnosed with P-SIRS and 30 clinically healthy controls. Retinal images were captured at admission using a smartphone coupled with a Volk iNView® (Volk®, Mentor, OH, USA) device. Retinal arteriolar and venular calibers within 0.5–1 disk diameter from the optic disk were measured using ImageJ software (https://imagej.net/ij/index.html). Concurrent hematology, serum biochemistry, VEGF, and IL-4 levels were analyzed. Statistical comparisons and correlations were assessed using non-parametric tests and Spearman’s correlation. Retinal arteriolar calibers were significantly narrower (p = 0.0001) and venular calibers significantly wider (p = 0.0068) in P-SIRS patients compared to controls. Serum VEGF and IL-4 concentrations were markedly elevated in the P-SIRS group (p < 0.05). Retinal venular calibers positively correlated with band neutrophils (p = 0.02), monocytes (p = 0.04), and negatively with albumin (p = 0.008). Retinal arteriolar calibers negatively correlated with segmented neutrophils (p = 0.04) and VEGF (p = 0.0003). No ophthalmoscopically visible retinal lesions were detected. Bitches with pyometra-induced SIRS exhibited significant retinal microvascular alterations characterized by arteriolar narrowing and venular dilation. These vascular changes correlated with systemic inflammatory markers and VEGF levels, suggesting that retinal vascular assessment may serve as a non-invasive biomarker for systemic inflammation in veterinary patients. Despite microvascular changes, no clinically visible retinal lesions were observed, warranting further longitudinal studies to elucidate their prognostic significance.
Research Article | 31 May 2025
Physiological, cytological, and reproductive hormone characterization of estrus in Pasundan heifers synchronized with double injection of prostaglandin F2α
Rini Widyastuti, Nena Hilmia, Diky Ramdani, Rahmat Hidayat, Iman Hernaman, Andre Rivanda Daud, Ken Ratu Ghazirah Alhuur, Matni Syifa Bayani, Rangga Setiawan, Sigit Prastowo, Santoso Santoso, and Vidi Wulandari

Volume-18 | Issue-5 | Article-27 | https://doi.org/10.14202/vetworld.2025.1357-1364

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Pasundan cattle, a native Indonesian breed with valuable reproductive traits, face population decline due to limited conservation efforts. Estrus synchronization using prostaglandin F2α (PGF2α) is a viable strategy to support genetic improvement and sustainable breeding. This study aimed to evaluate the physiological, cytological, and hormonal responses associated with estrus synchronization in Pasundan heifers following a double-injection protocol of PGF2α. Eighteen healthy Pasundan heifers (2.0–2.5 years old; body condition score 3) received two intramuscular PGF2α injections 11 days apart. Estrus signs were assessed through vulva morphology, cervical mucus viscosity, and vaginal electrical resistance (VER) on days 0, 4, 5, 11–15. Vaginal cytology was conducted to classify epithelial cells, and blood samples were analyzed for follicle-stimulating hormone (FSH), progesterone, and estradiol through enzyme-linked immunosorbent assay. Data were analyzed using one-way analysis of variance with Tukey’s post hoc test (p < 0.05). The peak estrus response occurred on day 14 post-initial injection, marked by maximal vulvar swelling (7.27 ± 1.15 cm), highest mucus viscosity (14.9 ± 3.00 mm), and lowest VER (198.67 ± 29.61 Ohms). Cytologically, superficial and keratinized epithelial cells dominated (64.22%), indicating estrus. Hormonal assays revealed elevated FSH (5.08 mIU/mL) and estradiol (0.214 pg/mL), alongside a nadir in progesterone (0.162 ng/mL). Estrus was observed in 88.89% of heifers on day 14. Double-injection PGF2α effectively synchronized estrus in Pasundan heifers, with day 14 being optimal for fixed-time artificial insemination (FTAI). The synchronization protocol demonstrated clear correlations between physical, cytological, and hormonal parameters. This protocol provides a reliable basis for reproductive management in Pasundan cattle, facilitating conservation and productivity. Future studies should assess conception outcomes post-FTAI to validate long-term reproductive efficiency.
Research Article | 31 May 2025
Invasion dynamics and ecological impacts of Anisakis typica in commercial fish from the Western Pacific Ocean
Dhito Dwi Pramardika, Fadjar Satrija, Sulistiono Sulistiono, Risa Tiuria, and Arifin Budiman Nugraha

Volume-18 | Issue-5 | Article-28 | https://doi.org/10.14202/vetworld.2025.1365-1376

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The Western Pacific Ocean hosts rich marine biodiversity, yet the parasitic infections affecting its commercial fish species remain underexplored. This study investigated the prevalence, intensity, morphological characteristics, molecular identity, and ecological impacts of Anisakis typica infection in commercial fish from this region. Between January and April 2024, 178 fish samples from 12 commercial species were collected across three geographical clusters: Sangihe Island, Kahakitang Island, and Marore Island. Fish specimens were morphologically identified and dissected for parasite detection. Morphological identification of larvae was complemented by molecular characterization through polymerase chain reaction amplification of the internal transcribed spacer (ITS)1–5.8S–ITS2 rDNA region, followed by sequencing and phylogenetic analysis. A. typica infection was observed in 10.7% of the fish sampled, with moderate infection intensity. Euthynnus affinis exhibited the highest prevalence (41.2%) and intensity (15.4). The intestinal wall was the dominant predilection site (96.3%). Molecular analyses confirmed A. typica infection in E. affinis, Katsuwonus pelamis, Decapterus kurroides, and Variola louti, with high genetic similarity (93.38%–100%) to isolates previously reported from China. Notably, this study provides the first documentation of A. typica infection in D. kurroides and V. louti. This study highlights the ecological and zoonotic significance of A. typica infection in commercially important fish from the Western Pacific Ocean. The findings underscore the potential threats to marine ecosystem stability, fishery sustainability, and public health. High genetic proximity between A. typica isolates from Indonesia and China suggests historical host migrations, emphasizing the need for regional surveillance and integrated management strategies. Enhanced inspection practices and public awareness initiatives are crucial to mitigate the zoonotic risks posed by consuming infected fish.
Systematic Review | 31 May 2025
Histological response and blood glucose level in a diabetic animal model after the oral administration of Mucuna pruriens: A systematic review and meta-analysis
Tri Wahyu Pangestiningsih, Dian Meididewi Nuraini, Morsid Andityas, and Ariana Ariana

Volume-18 | Issue-5 | Article-29 | https://doi.org/10.14202/vetworld.2025.1377-1388

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Mucuna pruriens (MP) has emerged as a promising natural antidiabetic agent due to its rich bioactive composition. Although numerous preclinical studies have reported its hypoglycemic and histological benefits, a comprehensive synthesis quantifying these effects has been lacking. This study systematically evaluated the dual impact of orally administered MP extract on histopathological changes and blood glucose levels in diabetic animal models through a systematic review and meta-analysis. A systematic literature search was conducted across four databases (PubMed, Scopus, ScienceDirect, and Google Scholar) without date restrictions. Eligible in vivo studies were selected based on predefined inclusion criteria, and data were extracted following PRISMA guidelines. Risk of bias was assessed using the systematic review center for laboratory animal experimentation tools. Histological outcomes were summarized descriptively, while blood glucose levels were analyzed quantitatively using a random-effects meta-analysis. Subgroup analyses were performed based on MP concentration, duration of administration, and plant part used Sixteen studies were included, with 13 eligible for meta-analysis. MP extract significantly reduced blood glucose levels, with an overall standardized mean difference of −18.36 (95% confidence intervals: −21.22, −15.51; p < 0.01). Subgroup analyses revealed that lower MP doses (≤100 mg/kg) achieved superior glycemic control with prolonged administration (>4 weeks), whereas higher doses (≥200 mg/kg) were most effective within 1–4 weeks. Histological analysis indicated regenerative effects of MP on the pancreas, liver, pituitary gland, and corpus cavernosum. Seed extracts exhibited a stronger hypoglycemic effect compared to leaf extracts. Potential publication bias was detected but was addressed through trim-and-fill analysis. MP extract demonstrates significant antidiabetic potential through glycemic regulation and organ tissue restoration. Lower concentrations are preferable for long-term administration, while higher concentrations are optimal for short-term therapy. The findings advocate MP as a valuable candidate for integrative diabetes management strategies. Further clinical studies are recommended to validate its translational potential.
Research Article | 31 May 2025
Mitochondrial markers (cytochrome c oxidase subunit I and 16S ribosomal RNA) as supporting biomarkers for wild bird identification
Julián Marín-Villa, Albeiro López-Herrera, Daisy A. Gómez-Ruiz, Diana C. Restrepo-Rodas, Geraldine Sánchez-Rodríguez, and Cristina Úsuga-Monroy

Volume-18 | Issue-5 | Article-30 | https://doi.org/10.14202/vetworld.2025.1389-1399

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Illegal wildlife trafficking is a critical threat to biodiversity, particularly in megadiverse countries such as Colombia. Birds, notably psittacines, are among the most targeted taxa. Morphological identification is often insufficient, especially when dealing with cryptic species or degraded samples. This study aimed to assess the utility of mitochondrial markers cytochrome c oxidase subunit I (COI) and 16S ribosomal RNA (16S rRNA) as molecular tools for species-level identification of psittacines housed at the Conservation Park of Medellín. Six adult psittacines from the genera Ara and Pionus were selected based on availability. Blood samples were collected and genomic DNA was extracted using a commercial kit. Polymerase chain reaction amplification of partial COI and 16S rRNA gene fragments was performed, followed by Sanger sequencing. Sequence identity was confirmed using BLASTn and the Barcode of Life Data System (BOLD). Phylogenetic relationships were analyzed using Neighbor-Joining, Maximum Likelihood, and Bayesian Inference approaches. Molecular results showed 100% concordance with prior morphological identification for all six individuals. COI and 16S rRNA sequences allowed clear species-level identification with similarity values >98%. Phylogenetic analyses for both markers yielded congruent tree topologies, with high branch support (>90%), further validating species identification. Maximum interspecific divergence for COI was observed between Ara macao and Pionus fuscus (0.15980), while 16S rRNA showed lower divergence values. All generated sequences were submitted to GenBank and BOLD in accordance with findable, accessible, interoperable, reusable principles. This study confirms the robustness of COI and 16S rRNA mitochondrial markers in accurately identifying psittacine species. The integration of molecular and morphological approaches enhances forensic investigations, facilitates biodiversity conservation, and contributes to efforts against wildlife trafficking. Expanding genetic databases for Neotropical avifauna, especially for commonly trafficked species, is imperative. Future research should adopt integrative genomic approaches involving nuclear markers to overcome the maternal inheritance limitation of mitochondrial DNA.