In The Gambia, malaria transmission persists due to insecticide resistance and residual vector behavior, despite extensive use of indoor residual spraying and insecticide-treated nets. Community-led larval source management using Bacillus thuringiensis var. israelensis (Bti) offers a sustainable vector control alternative. This study aimed to assess the effectiveness and feasibility of community-led Bti application for reducing Anopheles mosquito populations, compared to expert-supervised application and non-intervention control arms. A non-randomized controlled trial was conducted across malaria-endemic regions in The Gambia from 2023 to 2024. Intervention arms included: (1) community-led Bti application, (2) expert-supervised Bti application, and (3) untreated control. Trained volunteers and entomologists applied Bti to breeding sites at weekly or biweekly intervals. Entomological surveys were conducted biweekly to monitor larval, pupal, and adult mosquito densities. Data were analyzed using generalized linear mixed models and negative binomial regression, adjusting for environmental covariates. By round 10, community-led and expert-supervised interventions achieved 96.8% and 98.6% reductions in larval density, 97.4% and 99.1% reductions in pupal emergence, and 96.2% and 98.8% reductions in adult mosquito populations, respectively. Statistically significant declines in mosquito densities were observed by 2024 (p < 0.001). Community participation enabled high coverage and operational sustainability, with over 85% of participants reporting visible mosquito reduction. Seasonal Bti application, especially when led by trained community members, significantly suppresses Anopheles populations. Although expert-supervised methods yielded slightly higher efficacy, community-led biolarviciding offers a scalable, sustainable, and environmentally safe vector control strategy, supporting The Gambia’s malaria elimination goals.

Local Indonesian chickens possess valuable dual-purpose traits for both meat and egg production, but exhibit lower productivity compared to commercial breeds. Genetic enhancement through selective crossbreeding and molecular marker analysis, such as growth hormone (GH) gene polymorphism, offers a strategy to improve performance traits. This study aimed to characterize quantitative traits and analyze GH gene polymorphism in crossbred chickens resulting from mating Arab chickens with five indigenous breeds. Five local breeds, Kampung Super, Sentul, Bangkok, Kampung, and Merawang, were each crossed with Arabian chickens using a 1:7 male-to-female ratio. Phenotypic evaluations included body weight (BW), weight gain, and 18 morphometric traits measured at various ages. Egg traits were monitored for 4 weeks. GH gene polymorphism was identified in 500 crossbred individuals using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) with the AluI enzyme restriction. Data were analyzed through a one-way analysis of variance and General Linear Models to determine phenotypic and genotypic associations. Significant differences (p < 0.05) in BW, BW gain, morphometric size, and egg traits were observed among parental and crossbred groups. Kampung Super × Arab chickens showed the highest performance across growth and reproductive metrics. The highest weight gain occurred between 2 and 3 months of age. Back height emerged as a key morphometric indicator of growth differences. PCR-RFLP revealed GH gene polymorphism with three genotypes: (+/+), (+/−), and (−/−). The (+/+) genotype had a significant (p < 0.05) positive impact on BW, weight gain, and back height. All populations were in Hardy–Weinberg equilibrium, and polymorphic information content values (~0.437) indicated moderate genetic diversity. This study is the first comprehensive integration of morphometric, phenotypic, and GH genotypic data in Indonesian crossbred chickens. The findings support the implementation of marker-assisted selection to enhance growth traits in breeding programs. Future work should assess multi-generational effects and integrate additional molecular markers to optimize breeding strategies across tropical poultry systems.

Acinetobacter baumannii is a multidrug-resistant (MDR) pathogen notorious for its biofilm formation and persistence in clinical and veterinary settings. Its resistance is exacerbated by quorum sensing (QS) pathways that regulate virulence and biofilm maturation. Disrupting QS and biofilm integrity using plant-derived compounds presents a promising alternative to traditional antibiotics. This study aimed to evaluate the antibiofilm and anti-QS potential of Paederia foetida Linn. ethanolic extract against A. baumannii, integrating gas chromatography–mass spectrometry (GC-MS) profiling, molecular docking, and in vitro assays. Leaves of P. foetida were extracted with ethanol and analyzed by GC-MS to identify major bioactive constituents. Molecular docking was conducted against five QS and biofilm-associated A. baumannii proteins (AF-A0A7S8WE28-F1-v4, AF-A0A059ZL64-F1-v4, AF-Q2VSW6-F1-v4, AF-A0A2P1B9S4-F1-v4, and AF-A0A5P9VY74-F1-v4). Absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiles and drug-likeness of key compounds were assessed in silico. Antimicrobial activity was determined by broth microdilution (minimum inhibitory concentration [MIC]/minimum bactericidal concentration [MBC]), and biofilm inhibition was evaluated through crystal violet microtiter assays. Morphological damage was examined using field emission scanning electron microscopy (FE-SEM). GC-MS identified 30 phytoconstituents, with 5-hydroxymethyl-2-furaldehyde, 4H-pyran-4-one derivative, and eugenol as predominant compounds. Eugenol exhibited the highest binding affinity, particularly with AbaR (−6.3 kcal/mol). The extract showed significant antimicrobial activity (MIC = 7.81 mg/mL; MBC = 31.25 mg/mL) and dose-dependent inhibition of biofilm biomass (p < 0.001). FE-SEM imaging confirmed dose-responsive membrane damage and disruption of the biofilm. ADMET predictions revealed favorable oral bioavailability and low toxicity for selected compounds. P. foetida extract exhibits potent antibacterial, anti-QS, and antibiofilm activity against MDR A. baumannii, supported by its phytochemical diversity, favorable pharmacokinetics, and strong protein-ligand interactions. These findings suggest its promise as a plant-derived therapeutic aligned with the One Health framework to combat antimicrobial resistance in both human and veterinary medicine.

White spot syndrome virus (WSSV) is a devastating pathogen in shrimp aquaculture, with viral protein 28 (VP28) playing a critical role in host cell attachment and entry. The extracellular domain of VP28 (residues 35–95) is immunogenic and essential for infection; however, its receptor interaction mechanisms remain incompletely elucidated. This study aimed to evaluate the tissue-binding affinity of full-length VP28 and its derived peptides (P1: Residues 35–65; P2: Residues 66–95) as well as a multimeric chimeric virus-like particle (K5-VLP) displaying VP28 on the surface of Macrobrachium rosenbergii nodavirus capsids to enhance host tissue interaction. Recombinant VP28, synthetic peptides (P1, P2), and chimeric K5-VLP were produced and characterized. Binding and inhibition assays were performed using enzyme-linked immunosorbent assay and immunofluorescence microscopy on shrimp gill, hemocyte, muscle, stomach, and hepatopancreas tissues. Full-length VP28 exhibited strong binding to gill, hemocyte, and muscle tissues. The P1 and P2 peptides showed moderate binding compared to rVP28. Notably, K5-VLP demonstrated a 1.7-fold higher binding affinity than rVP28 in gill tissues and significantly outperformed P1 and P2 peptides. Inhibition assays confirmed that K5-VLP more effectively interfered with VP28 binding than peptides. Structural analysis and transmission electron microscopy confirmed correct assembly and surface presentation of VP28 on the VLPs. Multimeric display of VP28 on K5-VLP enhances its binding affinity to shrimp tissues compared to monomeric or peptide forms. This suggests a promising platform for antiviral strategies, including competitive inhibition of WSSV entry and targeted therapeutic delivery in shrimp aquaculture.

The small intestinal microbiota plays a pivotal role in poultry digestion and immune function. Rearing systems can influence their composition, thereby affecting the overall health and performance of the birds. This study aimed to investigate how rearing systems (intensive [IN] vs. free-range [FR]) and immune status, reflected by leukocyte profiles, influence the small intestinal microbiome of IPB-D3 chickens, a genetically improved Indonesian local breed. Ninety IPB-D3 chickens were reared for 12 weeks under either IN or FR systems. Hematological profiling was conducted to assess health status, with leukocyte counts used to stratify birds. Microbiota samples from the small intestine were analyzed using full-length 16S ribosomal RNA (V1–V9) sequencing on the Oxford Nanopore platform. Taxonomic identification was performed using the SILVA database. Statistical comparisons were made using t-tests, and microbial diversity was assessed through alpha and beta diversity metrics. While most hematological parameters did not differ significantly between rearing systems, total leukocyte counts were higher in intensively reared chickens (p = 0.002). FR chickens exhibited significantly greater microbial diversity (p < 0.05) across multiple alpha diversity indices. A total of 1,294 unique species were identified in FR birds versus 720 in the IN group, with 1,761 shared species. Leukocyte level further influenced microbial profiles; chickens with high leukocyte (HL) counts were dominated by Ligilactobacillus aviarius, whereas low-leukocyte chickens had a higher abundance of Bacteroides caecigallinarum. Gallibacterium anatis, a potential pathogen, dominated in IN systems with elevated leukocytes. This study demonstrates that both the rearing environment and immune status substantially influence small intestinal microbial composition in IPB-D3 chickens. FR systems promoted richer, more beneficial microbial communities, while IN systems, especially with HL levels, were associated with opportunistic pathogens. Leukocyte profiling may serve as a non-invasive biomarker for gut health, supporting future development of precision poultry management strategies and immune-responsive probiotics.

Coccidiosis is a widespread protozoan disease that severely impacts poultry health and productivity. The Qinghao Changshan (QHCS) formula, composed of multiple traditional Chinese medicinal herbs, is widely used in China for coccidiosis control. Despite its proven clinical efficacy, the molecular mechanisms underlying its therapeutic action remain poorly understood. This study aimed to elucidate the active components and molecular mechanisms of QHCS against coccidiosis using an integrated approach combining network pharmacology and molecular docking. Active compounds of QHCS were identified from public pharmacological databases based on criteria of oral bioavailability ≥ioa and drug-likeness ≥rug-l Targets of these compounds were predicted using SwissTargetPrediction and PharmMapper, and disease-related genes were retrieved from GeneCards, DrugBank, OMIM (Online Mendelian Inheritance in Man), and Therapeutic Target Database. Overlapping targets were visualized using Venn diagrams, and protein–protein interaction (PPI) networks were constructed using STRING and Cytoscape. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to explore relevant biological functions and pathways. Molecular docking was used to validate interactions between selected active compounds (isorhamnetin, kaempferol, quercetin) and key targets (epidermal growth factor receptor [EGFR], estrogen receptor 1 [ESR1], progesterone receptor [PGR]). Sixty-nine active compounds and 3476 potential targets of QHCS were identified, with 11 targets overlapping with 87 coccidiosis-related genes. Eight core targets–Amyloid Beta Precursor Protein, interleukin 6, TNF Receptor Associated Factor 1, Platelet Derived Growth Factor Receptor Beta, EGFR, ESR1, Erb-B2 Receptor Tyrosine Kinase 2, and PGR–were identified through PPI network analysis. GO and KEGG enrichment revealed key pathways including focal adhesion, calcium signaling, mitogen-activated protein kinase, ErbB signaling pathway, forkhead box O, and gap junction pathways. Molecular docking confirmed strong binding affinities of isorhamnetin, kaempferol, and quercetin to EGFR, ESR1, and PGR, supporting their regulatory roles in these signaling pathways. QHCS exhibits anti-coccidial activity by modulating multiple signaling pathways and molecular targets through its key bioactive constituents. These findings provide mechanistic insights into the therapeutic effects of QHCS and lay a theoretical foundation for its broader application in veterinary parasitology.

Tropical climates expose livestock to prolonged heat stress, leading to compromised health, growth, and productivity. Nutritional interventions using omega-3 fatty acids and antioxidants may improve resilience in such environments. This study evaluated the effects of lemuru fish oil (LO), vitamin E, and selenium – individually and in combination – on hematological, physiological, antioxidant, and growth parameters of Garut lambs exposed to tropical heat stress. Forty male Garut lambs (23.52 ± 2.51 kg; 9–10 months old) were randomly assigned to five dietary groups: Control (CNT), LO (6%, LO), LO + 500 IU Vitamin E, LO + 0.5 ppm selenium, and LO + both Vitamin E and selenium (LOES). The 60-day trial took place during the dry season under severe to extreme heat stress conditions (temperature-humidity index: 25.13–40.07). Parameters assessed included nutrient intake, physiological responses, blood hematology, antioxidant status, and growth performance. Supplementation with LO and antioxidants significantly improved hematological indicators (e.g., hemoglobin), enhanced antioxidant defenses (e.g., increased superoxide dismutase [SOD] and glutathione, reduced malondialdehyde [MDA], and cortisol levels), and stabilized physiological responses (e.g., lower rectal temperature and heart rate). The LOES group showed the most pronounced improvements: SOD increased by 107%, MDA decreased by 62%, and cortisol levels were reduced by 28% compared to the CNTs. However, no significant differences were observed in average daily gain or feed efficiency. The combination of LO with Vitamin E and selenium effectively mitigated heat-induced oxidative and physiological stress in Garut lambs. Although growth parameters remained unchanged, the improved physiological and antioxidant status suggests that these supplements may be valuable functional feed additives for enhancing animal welfare and resilience under heat stress. Further research is warranted to investigate the long-term effects on productivity and reproduction.

Triple-negative breast cancer (TNBC) presents therapeutic challenges due to its aggressive nature and lack of targeted treatments. Programmed death-ligand 1 (PD-L1) and interferon-gamma (IFN-γ) are key immune modulators in tumor immune evasion. Annonacin, a natural acetogenin from Annona species, has shown promising anticancer properties, though its immunomodulatory mechanisms remain underexplored. This study aimed to investigate the dual apoptotic and immunomodulatory effects of annonacin on PD-L1 and IFN-γ expression using combined molecular docking and in vitro assays in TNBC (4T1) cells. Molecular docking simulations were conducted to assess annonacin’s interaction with PD-L1 (Protein Data Bank [PDB] ID: 6PV9) and IFN-γ (PDB ID: 1FG9). In vitro experiments using 4T1 cells involved 3-(4,-5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays for cytotoxicity, Annexin V-fluorescein isothiocyanate staining for apoptosis, and flow cytometry to analyze PD-L1 and IFN-γ expression following treatment with annonacin (1.5–25 μg/mL). Docking scores indicated moderate binding affinities of annonacin to IFN-γ (–5.2 kcal/mol) and PD-L1 (–5.0 kcal/mol), involving both hydrogen bonds and hydrophobic interactions. Annonacin exhibited a selective cytotoxic effect on 4T1 cells with a half-maximal inhibitory concentration of 15 μg/mL and a selectivity index of 2.6. Apoptosis was induced in a concentration-dependent manner, with late apoptotic populations peaking at 25 μg/mL. PD-L1 and IFN-γ expression peaked at 6.25 μg/mL, followed by a decline at higher doses, suggesting a dose-dependent immunomodulatory shift from immune activation to suppression. Annonacin modulates immune checkpoint (PD-L1) and cytokine (IFN-γ) expression while promoting apoptosis in TNBC cells. These results highlight its potential as a dual-function anticancer agent, warranting further preclinical evaluation for use as a monotherapy or in combination with immunotherapies.

Dromedary camels exhibit unique immune adaptations that enable survival in harsh environments with high microbial exposure. However, the cellular mechanisms underpinning their innate immune responses, particularly oxidative respiratory bursts, remain underexplored. This study aimed to investigate the kinetics of reactive oxygen species (ROS) production in camel leukocytes in response to selected bacterial and fungal pathogens and to assess the effect of serum opsonization on ROS generation. Whole blood from six clinically healthy female dromedary camels was stimulated with opsonized and non-opsonized Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae (three strains), and Candida albicans. Luminol-enhanced chemiluminescence (CL) assays were used to quantify ROS production over a 3-h period. Colony-forming units were evaluated to confirm microbial viability post-opsonization. Key ROS metrics included area under the curve, peak emission (relative light unit), and time to peak. Camel neutrophils demonstrated distinct pathogen-specific ROS kinetics. P. aeruginosa and K. pneumoniae 1705 elicited the highest total ROS on serum opsonization, whereas S. aureus and C. albicans showed minimal ROS induction. E. coli failed to induce a measurable ROS response. Serum opsonization significantly enhanced total ROS production and shortened peak response time for K. pneumoniae strains. In contrast, it reduced total ROS output for S. aureus and C. albicans without significantly affecting their peak kinetics. This study provides the first comprehensive analysis of microbial-specific ROS production in camel whole blood using a luminol-based CL assay. The findings underscore the variability in camel innate immune responses to different pathogens and highlight the modulatory role of serum opsonization. These insights could inform future strategies in camel immunotherapy, vaccine development, and disease diagnostics.

Doxorubicin, a widely used chemotherapeutic agent, is associated with reproductive toxicity due to its induction of oxidative stress and testicular damage. Emerging evidence suggests that rosuvastatin and ramipril may possess antioxidant and cytoprotective properties beyond their conventional uses. However, their comparative efficacy in preventing doxorubicin-induced testicular toxicity remains unclear. This study aimed to evaluate and compare the protective effects of rosuvastatin and ramipril on testicular function, oxidative stress markers, and reproductive outcomes in a rat model of doxorubicin-induced testicular toxicity. Twenty-four male Wistar rats were randomly allocated into four groups: Control, doxorubicin-only, rosuvastatin + doxorubicin, and ramipril + doxorubicin. Doxorubicin (5 mg/kg, intraperitoneal) was administered on days 7, 14, and 21, while rosuvastatin or ramipril (5 mg/kg/day, oral) was given for 21 days. On day 45, evaluations included testicular index, sperm count and motility, serum testosterone levels, oxidative stress markers (malondialdehyde [MDA], nitric oxide [NO], glutathione [GSH]), and histopathological analysis using Johnsen scoring. Both rosuvastatin and ramipril significantly restored the testicular index compared to the doxorubicin group (p < 0.05). Ramipril markedly increased serum testosterone, GSH, and NO levels while reducing MDA. Sperm motility and count showed partial improvement, notably in the ramipril group. Histopathological alterations were attenuated in both treatment groups, with improved Johnsen scores and reduced architectural disruption. Ramipril and rosuvastatin mitigate doxorubicin-induced testicular toxicity through antioxidant mechanisms. Ramipril demonstrated superior efficacy in preserving reproductive hormone levels and sperm function. These findings highlight its potential as a fertility-protective agent during chemotherapy. Further long-term and mechanistic studies are warranted.

Lumpy skin disease (LSD) is a severe transboundary viral infection in cattle, caused by the LSD virus (LSDV), leading to economic losses in the livestock industry. Conventional live-attenuated vaccines face limitations such as strain recombination, incomplete protection, and adverse effects. Therefore, safer and more targeted vaccine strategies are urgently needed. This study aimed to design, simulate, and express a novel multi-epitope vaccine (MEV) candidate against LSDV using a computational immunoinformatic pipeline. Four immunogenic LSDV proteins – P35, A4L, A33R, and L1R – were selected based on their structural and antigenic significance. B- and T-cell epitopes were predicted and filtered using antigenicity, allergenicity, and toxicity criteria. Selected epitopes were linked using specific linkers and an adjuvant to construct an MEV. Molecular docking was performed with bovine toll-like receptors (TLRs), and stability was evaluated through molecular dynamic simulations (GROMACS and iMODS). Codon optimization and heterologous expression of the construct were performed in Escherichia coli using the pET-28a(+) vector. Expression was checked through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. A total of 23 epitopes from the four LSDV proteins were incorporated into a 514 amino acid-long vaccine construct. The designed construct demonstrated high antigenicity, non-allergenicity, solubility, and favorable physicochemical properties. Docking with bovine TLR4 revealed stable binding with significant interaction residues. Molecular dynamics confirmed structural stability over 50 ns simulations. The recombinant construct was successfully expressed as a ~59 kDa His-tagged protein in E. coli, confirmed by SDS-PAGE and Western blotting. This study demonstrates a comprehensive computational and experimental workflow for developing a multi-epitope subunit vaccine against LSDV. The MEV candidate shows strong immunogenic potential, structural stability, and recombinant expression feasibility, offering a promising alternative to traditional vaccines. Further in vivo evaluation is warranted to assess protective efficacy.

Avian influenza (AI) is a highly contagious zoonotic disease affecting birds and, occasionally, humans. Ecuador confirmed its first case of AI in late 2022, resulting in significant avian mortality and economic losses. In response, the Ecuadorian government implemented a mandatory control program emphasizing pre-vaccination diagnostics using reverse transcription quantitative polymerase chain reaction (RT-qPCR) or enzyme-linked immunosorbent assay. This study aimed to confirm the absence of AI virus type A in poultry across six major provinces of Ecuador following the 2022 outbreak, as part of the national eradication and vaccination campaign. A cross-sectional, retrospective molecular surveillance study was conducted from April 2023 to June 2024. A total of 343 pooled tracheal swab samples were collected from poultry farms in six provinces (Chimborazo, Cotopaxi, El Oro, Pastaza, Pichincha, and Tungurahua), covering over 1 million birds. The samples were analyzed using real-time RT-qPCR, targeting the M gene, and subtype-specific genes (H5, H7, and H7N9). All 343 pooled samples (100%) tested negative for AI virus type A, suggesting an absence of active viral circulation during the study period. The implementation of pre-vaccination testing and biosecurity protocols contributed to this outcome. The study confirms that AI virus type A was not circulating in Ecuadorian poultry during the surveillance period. These findings underscore the effectiveness of collaborative efforts among government, industry, and laboratories. Ongoing molecular surveillance remains essential for early detection and prevention of future outbreaks, reinforcing Ecuador’s commitment to One Health principles.

Coccidiosis, caused by Eimeria tenella, is a significant parasitic disease affecting poultry, resulting in severe intestinal damage and substantial economic losses. The increasing resistance to conventional anticoccidial drugs necessitates novel therapeutic strategies. This study aimed to synthesize and characterize a pyrazole-modified chitosan Schiff base–iron nanocomposite (ChSB-FeNPs) and evaluate its prophylactic and therapeutic effects against E. tenella in experimentally infected broiler chickens. ChSB-FeNPs were synthesized by incorporating iron nanoparticles into a pyrazole-modified chitosan Schiff base matrix and characterized using Fourier-transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscope (TEM) techniques. Sixty broiler chicks were randomly assigned to six groups: Uninfected controls, prophylactic and therapeutic ChSB-FeNPs treatments, and a standard amprolium treat-ment. Birds were infected with E. tenella and monitored over 28 days. Clinical signs, survival, body weight, feed conversion ratio (FCR), oocyst counts, lesion scores, liver enzyme activities (aspartate aminotransferase, alanine aminotransferase, and alanine aminotransferase), lipid profiles (low-density lipoprotein, high-density lipoprotein), and histopathological changes were assessed. ChSB-FeNPs-treated groups (both prophylactic and therapeutic) showed significantly reduced oocyst output, lesion scores, liver enzyme elevations, and histopathological damage compared to infected untreated controls. Prophylactic ChSB-FeNPs treatment notably improved body weight gain and FCR, with efficacy comparable to or exceeding that of amprolium. TEM confirmed the nanocomposite size (~39.5 nm), and cytotoxicity assays demonstrated safety at 0.133 μg/mL. ChSB-FeNPs exhibited potent anticoccidial effects, offering both preventive and therapeutic benefits against E. tenella infection in broilers. This nanocomposite represents a promising, next-generation alternative to conventional anticoccidial drugs, warranting further investigation for large-scale application.

Dengue is a significant mosquito-borne viral disease with increasing public health relevance, particularly in tropical and subtropical regions. Although it primarily affects humans, veterinary research plays a crucial role in understanding disease dynamics, particularly through the use of animal models and vector control strategies. This study aimed to analyze global research trends on dengue within the field of veterinary medicine between 2019 and 2024, identifying patterns in productivity, collaboration, and emerging thematic priorities. A bibliometric analysis was conducted using Scopus-indexed publications retrieved on December 15, 2024. The search included terms related to dengue and Aedes species within the veterinary subject area. Data were analyzed using SciVal for metrics and collaboration patterns and RStudio for statistical summaries and visua-lizations. Inclusion criteria encompassed peer-reviewed studies on dengue relevant to veterinary contexts published between 2019 and 2024. A total of 742 publications were identified across 67 journals, including 652 articles and 60 reviews. Annual output showed a 3.01% decline, with an average document age of 2.56 years and 9.0 citations per article. International collaboration was evident in 41.51% of publications, with Brazil, Argentina, and Australia emerging as key contributors. Thematic evolution demonstrated a shift from basic entomological studies (2019–2020) to applied control strategies, including Wolbachia-based mosquito interventions and citizen surveillance (2021–2022). The most prolific institutions were Fundação Oswaldo Cruz (Brazil) and the University of Florida (USA), while the journal Parasites and Vectors had the highest publication volume. This bibliometric review highlights the growing interdisciplinary nature of dengue research in veterinary medicine and the pivotal role of international collaboration. The observed decline in publication rate underscores the need to reinvigorate research efforts. The findings advocate for a One Health approach integrating veterinary, ecological, and public health disciplines to address vector-borne diseases. Future efforts should focus on enhancing global partnerships, standardizing animal models, and supporting innovative vector control strategies to mitigate the burden of dengue.

Bovine leukemia virus (BLV) is a globally distributed retrovirus that causes enzootic bovine leu-kosis, a chronic infection associated with significant economic losses in cattle. Conventional serological diagnostic tools such as agar gel immunodiffusion and enzyme-linked immunosorbent assay detect anti-BLV antibodies but cannot identify proviral DNA, especially in early infections or in calves with maternal antibodies. This study aimed to develop a sensitive and specific duplex quantitative polymerase chain reaction (qPCR) assay targeting the env gene of BLV with β-actin as an internal control and apply it for molecular surveillance and genotyping of BLV in cattle from six regions of Kazakhstan. A total of 1,680 bovine DNA samples from cattle aged over 3 years were collected from six administrative regions of Kazakhstan. A duplex qPCR assay was developed using primers targeting a conserved region of the BLV env gene and bovine β-actin. Sensitivity was assessed using plasmid and genomic DNA dilutions, and specificity was tested against existing WOAH-recommended and commercial polymerase chain reaction (PCR) protocols. Positive samples with cycle threshold <28 were subjected to nested PCR and Sanger sequencing for genotyping. Phylogenetic analysis was conducted using the maximum likelihood method. The developed qPCR assay demonstrated a sensitivity of 20 plasmid copies for the env gene and 6 genomic equivalents for β-actin per reaction, with high specificity comparable to international standards. The overall BLV infection rate was 38.9%, ranging from 13% in Pavlodar to 60.5% in East Kazakhstan. Among 149 sequenced positive samples, four genotypes (G1, G4, G7, and G8) were identified. Genotype G4 was predominant, comprising 79.2% of sequences and present in all six regions. The duplex qPCR assay is a robust, sensitive, and cost-effective diagnostic tool for detecting BLV provirus, including in animals with maternal antibodies or early-stage infections. The regional genotypic distribution underscores the need for tailored control strategies. This molecular surveillance provides essential baseline data for national BLV eradication programs and contributes to global BLV epidemiological mapping.

Mangosteen (Garcinia mangostana) and turmeric (Curcuma longa) are medicinal plants with well-documented antimicrobial, antioxidant, and anticancer properties, attributed to α-mangostin and curcumin, respectively. While their individual bioactivities are recognized, their synergistic potential and standardization through analytical validation remain underexplored, particularly in veterinary and pharmacological applications. This study aimed to (1) evaluate the synergistic antibacterial, antioxidant, and anticancer effects of ethanolic extracts of mangosteen pericarp and turmeric rhizome and (2) develop and validate a high-performance liquid chromatography (HPLC) method for the simultaneous quantification of their major bioactive compounds. Ultrasound-assisted extraction was employed to prepare ethanolic extracts. Antibacterial activities were assessed using disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration assays against five bacterial strains, with synergy evaluated through checkerboard fractional inhibitory concentration index. Antioxidant activity was measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging and total phenolic content (TPC). Cytotoxicity was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays on human hepatocellular carcinoma cells (HepG2), human breast adenocarcinoma cells (MCF-7), and human normal fibroblast cells. A reversed-phase HPLC method was developed and validated to simultaneously quantify α-mangostin and curcumin in the combined extract. Mangosteen and turmeric extracts showed MICs of 3.12 and 31.25 μg/mL, respectively. Combined extracts demonstrated additive or synergistic effects against Gram-positive bacteria and enhanced the efficacy of gentamicin (up to 19-fold MIC reduction). The combined extract exhibited the strongest antioxidant activity (half-maximal inhibitory concentration = 5.78 μg/mL) and highest TPC (1227.38 mg gallic acid equivalent/g extract). Cytotoxic assays revealed selective inhibition of HepG2 and MCF-7 cells, with no toxicity toward normal fibroblasts. The validated HPLC method enabled accurate, simultaneous quantification of curcumin (73.23 mg/g extract) and α-mangostin (146.80 mg/g extract) with excellent linearity (R2 > 0.9995) and recovery (99.08%–104.72%). The combination of mangosteen and turmeric extracts exhibits potent, selective, and synergistic antibacterial and anticancer properties, along with enhanced antioxidant capacity. The validated HPLC method provides a reliable tool for quality control and standardization of such polyherbal formulations, supporting their potential for therapeutic and veterinary applications.

Feline chronic gingivostomatitis (FCGS) is a debilitating inflammatory condition of the oral cavity in cats, associated with chronic pain, salivation, halitosis, and reduced quality of life. Omega-3 polyunsaturated fatty acids (n-3 PUFAs) have known anti-inflammatory properties and may offer a supportive treatment option. This study aimed to evaluate the safety and clinical efficacy of two marine-derived n-3 PUFA sources – krill oil and green-lipped mussel (GLM) oil – administered through lickable cat treats in cats with FCGS. Sixteen FCGS cats were randomized into three double-blinded treatment groups to receive daily lickable treats: Control (no n-3 PUFA), krill oil (100 mg), or GLM oil (100 mg) for 28 days. Clinical outcomes (stomatitis and pain scores, halitosis, and salivation) were assessed on days 0, 14, and 28 by veterinarians. Owner-reported outcomes and inflammatory cytokine levels (interleukin 6, interleukin 1 beta, and interferon-gamma) were also evaluated. Both krill and GLM oil treatments were well-tolerated, with no adverse changes in body weight, hematological, or biochemical parameters. Mean stomatitis and pain scores showed a non-significant downward trend in all groups. Halitosis and salivation improved in seven cats, particularly in the GLM and control groups. Owner-reported improvement was observed in 10 of 16 cats, with the highest in the krill oil group (75%). Cytokine levels demonstrated high variability and no statistically significant changes. No significant differences were observed between the n-3 PUFA sources. Daily administration of n-3 PUFA-enriched cat treats is safe and may provide mild clinical benefit in cats with FCGS, particularly in alleviating oral discomfort. While no significant differences were found between krill and GLM oils, both formulations were palatable and suitable as adjunctive therapy. Further research with larger cohorts, extended durations, and sensitive scoring systems is warranted to optimize treatment protocols.

The reproductive efficiency of livestock, especially indigenous breeds such as Donggala bulls, is pivotal to successful breeding programs. While conventional semen parameters are widely used, molecular markers, such as sperm protein profiles and DNA integrity, are emerging as reliable indicators of fertility. This study aimed to characterize the sperm protein profiles of Donggala bulls and examine their correlation with sperm DNA integrity and protamine deficiency. Frozen semen samples were collected from six Donggala bulls (aged 5–7 years). Pre-freezing evaluations included progressive motility (via computer-assisted sperm analysis), sperm morphology (using Diff-Quik staining), DNA integrity (assessed by the acridine orange assay), and protamine deficiency (assessed by the chromomycin A3 assay). Protein concentrations were determined using the bicinchoninic acid assay, and protein profiling was performed using 1D sodium dodecyl sulfate polyacrylamide gel electrophoresis. Band intensities and distributions were analyzed using ImageJ. Statistical correlations were analyzed using a one-way analysis of variance and Pearson’s correlation coefficients. Significant individual variation was observed in semen quality among bulls. Progressive motility ranged from 38.3% to 46.1%, DNA integrity from 79.5% to 96.8%, and protamine deficiency from 96.0% to 98.7%. The number of protein bands per sample varied between 8 and 11, with molecular weights ranging from 5 to 175 kilodaltons (kDa). Protein concentration ranged from 8.32 to 20.70 μg/mL. A 35 kDa protein band was notably absent in one bull, which may be linked to lower motility. Strong correlations were observed between sperm motility and DNA fragmentation (r = 0.628), protamine deficiency (r = 0.539), protein concentration (r = 0.658), and protein band expression (r = 0.788). Sperm protein profiles in Donggala bulls are significantly correlated with DNA integrity and protamine deficiency, indicating their potential as molecular biomarkers for fertility prediction. These findings provide a foundation for integrating protein profiling into breeding soundness evaluations, suggesting that targeted proteomic analysis may enhance reproductive management strategies.