Antimicrobial resistance (AMR) in enteric pathogens such as Escherichia coli and Salmonella spp. has emerged as a critical global health challenge affecting both human and animal populations. The widespread use of antibiotics in food-producing animals for therapeutic, prophylactic, and growth-promoting purposes has accelerated the selection and dissemination of resistant bacteria and resistance genes throughout the food chain. Animal-origin foods, including meat, milk, eggs, and fish, serve as important vehicles for the transmission of multidrug-resistant organisms and AMR genes to humans, repre­senting a significant One Health concern. This review provides an overview of the occurrence, molecular mechanisms, and transmission pathways of AMR in E. coli and Salmonella isolated from animal-derived foods. Common resistance determi­nants include β-lactamase genes (blaTEM and blaCTX-M), tetracycline resistance genes (tetA and tetB), and plasmid-mediated quinolone resistance genes, which facilitate horizontal gene transfer through plasmids, integrons, and transposons. Global surveillance reports from World Health Organization’s Global Antimicrobial Resistance Surveillance System, European Food Safety Authority, and World Organization for Animal Health reveal significant regional disparities, with limited monitoring capacity in Central Asia, Africa, and Latin America. Data from Kazakhstan indicate a high prevalence of multidrug-resistant E. coli and Salmonella in poultry, dairy, and cheese products, underscoring the urgent need for harmonized national sur­veillance and risk management strategies. The review also discusses alternative approaches to reduce antibiotic use in livestock production, including bacteriophage therapy, probiotics, phytogenic feed additives, vaccination, and nanotech­nology-based interventions. While these strategies show promising results in laboratory and pilot studies, their practical application remains constrained by regulatory, economic, and field validation challenges. An integrated One Health strategy, combining surveillance, antimicrobial stewardship, and non-antibiotic interventions, is crucial to mitigating the dissemina­tion of AMR along the farm-to-fork continuum. Strengthening laboratory networks, enhancing data sharing, and promoting collaboration among veterinary, environmental, and public health sectors will be crucial to safeguard food safety and global health security.

Canine mammary tumors (CMTs) serve as valuable comparative models for human breast cancer (HBC) owing to their shared biological and molecular features. However, well-defined cell lines representing the luminal B subtype remain limited. This study aimed to establish and characterize a novel CMT cell line, designated CMT-622, to expand available in vitro models for luminal B breast cancer research. Primary tumor tissue was collected from an 11-year-old female dog diagnosed with high-grade mammary carcinoma (T3N1M0). Tumor cells were isolated using enzymatic digestion and differential adhesion. Morphological, cytogenetic, and immunophenotypic characteristics were assessed using hematoxylin-eosin staining, immu­nohistochemistry, and immunofluorescence. Growth kinetics, clonogenicity, and chromosomal analyses were performed, and tumorigenicity was evaluated through xenograft assays in nude mice. Drug sensitivity and apoptosis were compared with two existing CMT lines (CMT-1211 and CMT-n7) using the cell counting kit-8 (CCK)-8 assay and flow cytometry. CMT-622 cells maintained stable proliferation beyond 40 passages with a doubling time of 46.23 h and >15% clon­ing efficiency. Karyotyping revealed hyperdiploidy (80–110 chromosomes; modal = 87). Immunohistochemistry and immu­nofluorescence confirmed estrogen receptor (+), progesterone receptor (–), and human epidermal growth factor receptor 2 (weak +) expression, consistent with a luminal B phenotype. Co-expression of cytokeratin-18 and vimentin indicated a par­tial epithelial–mesenchymal transition (EMT) state. In nude mice, CMT-622 exhibited moderate tumorigenicity and pulmo­nary metastasis. The line showed intermediate osthole sensitivity (half-maximal inhibitory concentration = 50.48 μM) and an apoptosis rate of 21%, between CMT-1211 and CMT-n7, indicating balanced proliferative and drug-responsive behavior. CMT-622 represents a newly established luminal B CMT cell line with stable growth, EMT plasticity, and mod­erate drug sensitivity, reflecting clinically relevant tumor aggressiveness. Its molecular and phenotypic consistency across in vitro and in vivo models underscores its reliability for translational oncology applications. CMT-622 provides a robust pre­clinical platform for exploring tumorigenesis, metastasis, and therapeutic responses in both veterinary and HBC contexts, bridging comparative and translational cancer research.

Acanthamoeba spp. is free-living protozoa capable of causing severe infections, notably Acanthamoeba keratitis, which is difficult to manage due to cyst resistance and the cytotoxicity of current treatments. Plant-derived com­pounds represent a promising alternative strategy. This study investigated the amoebicidal, anti-adhesive, and cytotoxic properties of Mangifera indica L. (mango) leaf extract against ocularly relevant Acanthamoeba spp. Crude ethanolic leaf extract of M. indica was prepared and evaluated against Acanthamoeba polyphaga American Type Culture Collection (ATCC) 30461 and Acanthamoeba castellanii ATCC 50739. Minimum inhibitory concentration (MIC) and minimum parasiticidal concentration were determined for trophozoites and cysts. Morphological changes were analyzed by scanning electron microscopy (SEM). Anti-adhesion assays were conducted using polystyrene surfaces, with a commercial multipurpose contact lens (CL) solution as a control. Cytotoxicity was tested in Vero cells using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay to establish the minimum cytotoxic concentration. The extract inhibited trophozoite growth at 2 mg/mL and demonstrated cysticidal activity at 4 mg/mL for A. polyph­aga and 32 mg/mL for A. castellanii. SEM revealed disruption of trophozoite morphology, loss of acanthopodia, and sur­face perforations in cysts. At MIC levels, adhesion was reduced by >70%, and even at 1/8 MIC, inhibition remained above 50%, comparable to a commercial multipurpose solution. Cytotoxicity assessment showed >80% Vero cell viability at 0.125 mg/mL, indicating a favorable therapeutic window. This is the first report demonstrating amoebicidal and anti-adhesive effects of M. indica L. leaf extract against ocular Acanthamoeba species. The dual trophozoiticidal and anti-adhesive actions, combined with low cytotoxicity, high­light its potential for development as a plant-based therapeutic agent, particularly in ocular formulations or CL disinfectants. Future work should focus on phytochemical isolation, mechanistic studies, and novel delivery systems to enhance efficacy and safety.

Mitigating enteric methane emissions in ruminants remains a global challenge in achieving sustain­able livestock production. Although seaweed supplementation has shown promising results, most research has focused on temperate species, leaving tropical species underexplored. This study investigated the nutritional composition, bioactive compounds, and in vitro rumen fermentation characteristics of two tropical seaweeds, brown seaweed (Sargassum binderi) and green seaweed (Kappaphycus striatum), as potential functional feed additives for ruminants. The proximate composition, macro- and micro-minerals were determined using Association of Official Analytical Chemists and Inductively Coupled Plasma–Optical Emission Spectrometry methods. In vitro digestibility of dry matter digestibility (DMD) and organic matter digestibility (OMD) was evaluated using the Tilley and Terry two-stage technique. Rumen fermentation characteristics, pH, ammonia (NH3), and total volatile fatty acids (VFA), were analyzed after 48 h of incubation. Amino acids and fatty acids were profiled using high-performance liquid chromatography and gas chromatography–flame ionization detection, respectively, while bioactive metabolites were identified through liquid chro­matography–high-resolution mass spectrometry metabolomics. Green seaweed exhibited a higher crude protein content (7.52%) and digestibility (DMD = 73.56%; OMD = 72.71%) than brown seaweed (6.84%; 46.38%; 44.99%). VFA production (136.75–151.75 mM) and NH3 concentrations (22.21–26.78 mM) differed significantly (p < 0.01) between species, while pH remained within the optimal range (7.00–7.21). Both seaweeds contained balanced essential and non-essential amino acid profiles and abundant polyunsaturated fatty acids, notably linoleic, α-linolenic, eicosapentaenoic acid, docosahexaenoic acid, and conjugated linoleic acid. Metabolomic screening identified ~85 bioactive compounds, including lipid-derived metabolites, amino alcohols, vitamins, and osmolytes such as betaine and cholecalciferol, indicating their potential to modulate rumen fermentation and enhance animal resilience. Both S. binderi and K. striatum demonstrated promising nutritional and bioactive potential as ruminant feed additives. Their compositional diversity suggests species-specific applications – S. binderi as an energy-dense supplement and K. striatum as a functional additive for stress adaptation. However, further in vivo trials are necessary to determine optimal inclusion levels, long-term safety, and methane mitigation efficacy under production conditions.

Zoonotic intestinal parasites remain a neglected public health problem in low-resource coastal com­munities where humans, free-roaming dogs, and contaminated environments interact closely. Cartagena, Colombia, lacks updated epidemiological data on intestinal parasitosis despite its high tourist influx and vulnerable populations. This study applied a One Health framework to investigate the prevalence of intestinal parasites in humans, dogs, and soil in two coastal regions of Cartagena (La Boquilla and Punta Arenas) and to identify associated risk factors and clinical manifestations. A cross-sectional survey was conducted between March 2024 and March 2025. Stool samples from 33 residents and 42 dogs were analyzed by direct microscopy with saline and Lugol’s solutions, while 78 soil samples were assessed using the Krumbein, Sloss, and Willis techniques. Dogs suspected of Dirofilaria spp. infection were additionally screened by the Woo test. Sociodemographic data, nutritional status, and clinical information were collected. Statistical analyses included descriptive tests, Chi-square/Fisher’s tests, logistic regression, and odds ratio (OR) estimation at a significance level of p ≤ 0.05. Soil samples showed high contamination with Toxocara spp. (46.2%), Strongyloides spp. (28%), and Ancylostoma spp. (25.7%). Among humans, 60.97% were positive for parasites, with Giardia spp. (15.15%), Entamoeba histolytica/dispar (12.12%), Ascaris lumbricoides (12.12%), and Enterobius vermicularis (12.12%) being the most frequent. In dogs, 33.33% carried intestinal parasites, predominantly Ancylostoma spp. (14.29%) and Giardia spp. (7.14%). Clinical manifestations in humans included loss of appetite, cough, dermatitis, and weight loss, while dogs frequently presented with pallor, dermato­logical lesions, and gastrointestinal signs. Logistic regression analysis indicated that a lack of canine deworming significantly increased the risk of human parasitic infections (OR: 3.80; 95% confidence interval: 0.98–14.66; p = 0.048). This One Health investigation highlights significant zoonotic risk from shared parasitic infections in humans, dogs, and contaminated soils in Cartagena’s coastal regions. The lack of systematic deworming and vaccination in dogs, poor sanitation, and close human–dog contact amplify transmission. Strengths of this study include its integrative human–ani­mal–environment approach, while limitations involve modest sample size and lack of molecular genotyping. Future work should apply molecular epidemiology to confirm cross-species transmission. Practical implications emphasize the urgent need for mass deworming campaigns, improved waste management, vector control, and public health education to reduce zoonotic intestinal parasite burden in vulnerable coastal communities.

Toxoplasma gondii is an obligate intracellular protozoan responsible for reproductive losses in sheep and significant zoonotic transmission to humans. Despite its known presence in Algeria, regional epidemiological data remain inconsistent. This study aimed to determine the seroprevalence of T. gondii and identify associated risk factors among aborted ewes in Tebessa Province, northeastern Algeria, within a One Health framework. A cross-sectional survey was conducted from September 2019 to October 2020 across three communes (Tlidjen, Al Ater, and Negrine). Serum samples (n = 297) were collected from recently aborted Ouled Djellal ewes aged 2–5 years. Anti-T. gondii immunoglobulin G antibodies were detected using the Toxo-Screen DA direct agglutination test at a 1:40 dilution. Risk factors, including location, parity, gestational stage, farming system, and presence of carnivores, were analyzed by χ² tests and multivariate logistic regression using R v4.0.3. The overall seroprevalence was 48.48% (144/297). Significant differences were observed across communes (Tlidjen 59.03%, Al Ater 42.72%, Negrine 30.00%; p < 0.001). Higher prevalence was recorded in sedentary (58.54%) than transhumant (26.09%) systems (odds ratio [OR] = 5.28; 95% confidence interval [CI]: 2.83–9.85; p < 0.001) and in farms with carnivores (63.31% vs. 28.91%; OR = 2.90; p < 0.001). Multiparous ewes were less likely to be seropositive than primiparous ones (OR = 0.55; p = 0.047). No significant association was found for gestation stage (OR = 1.58; p = 0.111). The high seroprevalence of T. gondii in aborted ewes indicates active environmental transmission and considerable reproductive and zoonotic risks in Tebessa. Strengthened farm biosecurity, feline population management, and public awareness of meat hygiene are urgently needed. Integrating veterinary, environmental, and public health surveillance will improve toxoplasmosis control within the One Health framework.

Duck circovirus (DuCV) is an immunosuppressive pathogen linked to poor growth, feather abnormalities, and increased susceptibility to co-infections, leading to significant economic losses in duck production. Rapid and large-scale serological screening tools are essential for epidemiological surveillance and biosecurity. This study aimed to develop and validate an indirect enzyme-linked immunosorbent assay (iELISA) based on a recombinant capsid (Cap) protein for sensitive and specific detection of antibodies against DuCV. The cap gene from a Thai DuCV genotype I isolate was cloned into the pQE-31 vector and expressed in Escherichia coli M15. The 27 kDa recombinant Cap protein was purified under denaturing conditions, and its antigenicity was confirmed by Western blotting. The iELISA was optimized by checkerboard titration to determine the optimal antigen coating concentration and serum dilution. Diagnostic sensitivity, specificity, cross-reactivity, repeatability, reproducibility, and agreement with Western blotting were assessed using 80 positive, 103 negative, and 189 field serum samples. The optimized iELISA used 12 μg/well of antigen and a 1:20 serum dilution, producing the highest positive-to-negative optical density ratio. Receiver operating characteristic curve analysis yielded an area under the curve of 0.996, with 97.5% sensitivity and 98.1% specificity. No cross-reactivity was detected with sera positive for duck Tembusu virus, duck viral enteritis virus, or Riemerella anatipestifer. Intra- and inter-assay coefficients of variation were below 6.5% and 9.1%, respectively. Diagnostic agreement with Western blotting across 189 field sera was 91.0%, with a Cohen’s kappa of 0.752, indicating substantial concordance. The developed recombinant Cap-based iELISA provides a reliable, specific, and reproducible tool for large-scale DuCV serosurveillance. Its high diagnostic accuracy and scalability support its application in flock-level monitoring, pre-movement screening, and epidemiological studies, facilitating improved biosecurity and informed disease control strategies within the duck industry.

The gut microbiota of broilers plays a pivotal role in nutrient absorption, immune modulation, and mineral metabolism. Feed additives can influence these microbial and physiological processes, yet their integrated effects remain insufficiently understood. This study aimed to intelligently evaluate the impact of various feed additives on the intestinal microbiota and mineral composition of broiler chickens and to develop machine learning (ML) models for clustering and classification of diet-associated mineral and microbial profiles. A total of 385 Arbor Acres broilers (7 days old) were allocated into 11 groups, including one control semi-synthetic diet (SSD), one group with a semi-synthetic deficient diet (SSDD), and nine experimental groups receiving SSDD with different additives: Probiotics (Soya-bifidum and Sporobacterin), dietary fibers (cellulose, lactulose, and chitosan), enterosorbents (enterosgel and activated carbon), and ultrafine particles (UFPs) (Cu and Fe). Microbiota composition was assessed by 16S ribosomal RNA sequencing, and body mineral composition was determined through inductively coupled plasma mass spectrometer. To overcome data scarcity, synthetic records were generated using conditional tabular generative adversarial networks. K-means and hierarchical agglomerative clustering were used for mineral profile grouping, while logistic regression, SVM, and decision tree models classified diet types. Hierarchical clustering revealed six distinct mineral profile groups (Silhouette = 0.524), with SSD and SSDD forming separate clusters. Feed additives such as UFPs, chitosan, and activated carbon induced similar mineral patterns. Key differentiating biomarkers were cobalt, zinc, strontium, arsenic, and lithium (p < 0.05). The decision tree classifier achieved 74% accuracy in predicting diet types based on microbiota data. Alpha diversity analysis showed enhanced microbial richness in groups fed lactulose, enterosgel, cellulose, or activated carbon. ML effectively elucidated complex relationships between diet, microbiota composition, and mineral metabolism in broilers. The integration of clustering and predictive models demonstrates the feasibility of intelligent feeding systems tailored to optimize gut health and nutrient utilization. Future studies integrating multi-omics data and broader farm-level validation will strengthen precision nutrition frameworks for sustainable poultry production.

Campylobacter jejuni and Campylobacter coli are leading causes of bacterial foodborne gastroenteritis worldwide, with poultry serving as a principal reservoir. The rapid emergence of antimicrobial-resistant (AMR) Campylobacter strains poses a growing public-health challenge, especially in developing countries where therapeutic options are limited. This study investigated the fecal carriage and AMR profiles of zoonotic Campylobacter species (ZCS) isolated from broilers. A cross-sectional study was conducted between February and July 2024. A total of 370 broiler fecal samples were collected using systematic random sampling and cultured on Modified Charcoal Cefoperazone Deoxycholate Agar. Phenotypic identification was performed by colony morphology, Gram staining, and biochemical tests. Antimicrobial susceptibility was assessed using the Kirby–Bauer disk diffusion method against nine antibiotics from distinct classes. Multiple antibiotic resistance (MAR) indices were calculated, and Fisher’s exact test was applied to determine statistical associations (p < 0.05). Campylobacter spp. were detected in 20% (74/370) of samples, comprising C. jejuni 6% (22/370) and C. coli 14% (52/370). Nearly all isolates (97.3%) exhibited multidrug resistance (MDR), with MAR indices ranging from 0.2 to 1.0 (mean = 0.8). Thirteen distinct AMR patterns were observed; seven were associated with C. coli and six with C. jejuni. The three most effective antibiotics were gentamicin (GEN) > ciprofloxacin > tetracycline (TET), though C. coli isolates were significantly more resistant to GEN (p = 0.001) and TET (p = 0.018). The 20% fecal carriage of ZCS in slaughtered broilers and the 97.3% MDR prevalence pose a serious public-health and food-safety threat. Prudent antimicrobial use strengthened farm biosecurity, and active AMR surveillance under a One Health framework are urgently needed to curb the spread of antibiotic-resistant Campylobacter spp. in poultry production systems and to safeguard human health.

Haemonchus contortus is a highly pathogenic blood-feeding nematode of small ruminants, responsible for severe anemia, production losses, and mortality. Excessive use of synthetic anthelmintics, especially benzimidazoles, has led to widespread drug resistance, prompting a need for alternative therapeutics. Nicotiana tabacum (tobacco) contains bioactive alkaloids such as nicotine, which target nematode acetylcholine receptors and may provide sustainable parasite control. This study aimed to isolate, characterize, and evaluate the in vitro and in vivo anthelmintic efficacy and safety of alkaloid-rich fractions of N. tabacum against benzimidazole-resistant H. contortus in goats, in line with the One Health approach and the United Nations Sustainable Development Goals (SDGs 2, 3, 12, and 15). Leaves of N. tabacum were authenticated and subjected to sequential solvent extraction. Alkaloid fractions were confirmed by TLC and HPLC. In vitro assays, including adult motility and egg hatch tests, were conducted at concentrations of 1–5 mg/mL against resistant H. contortus isolates. The most active fraction (ethyl acetate) was administered orally in naturally infected Beetal goats (n = 25) at low (0.8 mg/kg), medium (1.2 mg/kg), and high (1.6 mg/kg) doses for 14 days, alongside negative (saline) and positive (oxfendazole 4.5 mg/kg) controls. Fecal egg counts, hematology, and liver enzyme levels were analyzed to determine efficacy and safety. The ethyl acetate fraction exhibited complete (100%) adult worm mortality at 3–5 mg/mL and total egg-hatch inhibition at 4–5 mg/mL (p < 0.05). The LD₅₀ for adult worm mortality was 0.323 mg/mL. In vivo, the high-dose group (1.6 mg/kg) achieved a 76.2% fecal-egg-count reduction, exceeding oxfendazole (69.7%). No significant changes in alanine aminotransferase or aspartate aminotransferase were observed (p > 0.05), confirming hepatic safety, while serum proteins and red-blood-cell indices improved significantly (p < 0.05). Purified alkaloid fractions of N. tabacum, particularly the ethyl acetate extract containing nicotine, demonstrated strong, dose-dependent anthelmintic activity, and safety against benzimidazole-resistant H. contortus. These findings support N. tabacum as a sustainable, plant-based alternative to synthetic anthelmintics. The work advances the One Health framework and contributes directly to SDG 2 (Zero Hunger), SDG 3 (Good Health and Well-being), SDG 12 (Responsible Consumption and Production), and SDG 15 (Life on Land).

Conventional egg-yolk and milk-based extenders are widely used for semen cryopreservation but pose biosafety concerns and compositional variability that compromise standardization. Liposome technology offers a biosecure, uniform alternative. This study aimed to develop soy lecithin-derived nanoliposomes (NLs) using an ultrasonic-based process and to evaluate their efficacy as a Tris-based extender for Ongole-grade bull semen cryopreservation. Soy lecithin NLs were prepared through probe ultrasonication (15–45 min) and ultracentrifugation, followed by physicochemical characterization using particle size analysis, scanning electron microscopy (SEM), and high-resolution transmission electron microscopy. Tris-NL (TNL) extenders were formulated at concentrations of 5–25 mg/mL and compared with a Tris–egg-yolk (TEY, 20%) control. Fresh semen from five Ongole-grade bulls was evaluated for motility, viability, and morphology before and after freezing. Kinematic parameters were assessed through computer-assisted semen analysis, plasma membrane integrity by hypoosmotic swelling test, acrosome integrity using fluorescein isothiocyanate-conjugated peanut agglutinin/propidium iodide staining, and DNA fragmentation by Halomax-sperm chromatin dispersion assay. Optimized sonication (45 min) produced stable NLs (mean diameter 76 nm, zeta potential −43.2 mV) with uniform spherical morphology. Among the tested formulations, TNL 5–10 mg/mL showed significantly higher (p < 0.05) post-equilibration motility (up to 98%), progressive motility, and kinematic parameters (velocity curved line, velocity average path, and velocity straight line) than TEY. Post-thaw evaluations demonstrated improved sperm viability (≈66%), reduced abnormalities (<7%), enhanced plasma-membrane and acrosomal integrity, and lower DNA fragmentation (~1.2%) in the 5–10 mg/mL groups. SEM confirmed smoother sperm surfaces with minimal cryo-damage compared with TEY. Soy lecithin-derived NLs at 5–10 mg/mL serve as an effective and biosecure substitute for egg yolk in Tris extenders, enhancing motility, viability, and structural integrity of Ongole-grade bull spermatozoa. This locally developed, plant-based nanotechnology supports biosafety, import substitution, and sustainability of artificial insemination programs in Indonesia.

Palm kernel meal (PKM), a major by-product of the palm oil industry, is rich in nutrients but poorly utilized in poultry feed due to its high fiber and mannan content. Improving PKM digestibility through microbial bioconversion could reduce dependency on expensive protein sources, such as soybean meal. This study aimed to evaluate a consortium of Bacillus subtilis and Lactobacillus fermentum for its enzymatic activity, probiotic properties, and potential to enhance PKM utilization in poultry diets. The research was performed in four stages: (1) measurement of cellulase, mannanase, and protease activities in individual and combined bacterial cultures (seven treatments, five replications); (2) determination of enzyme activities in B. subtilis and L. fermentum (1:1) grown in de Man, Rogosa, and Sharpe broth supplemented with 0%–20% PKM (four treatments, seven replications); (3) in vitro probiotic characterization, including acid and bile tolerance, hydrophobicity, autoaggregation, coaggregation, and pathogen inhibition; and (4) evaluation of enzyme activity in natural media composed of coconut water and shrimp wastewater. Data were analyzed using analysis of variance and Duncan’s multiple range tests at p < 0.05. The 1:1 consortium exhibited the highest enzyme activities; cellulase (13.71 U/mL), mannanase (17.05 U/mL), and protease (9.32 U/mL). The consortium retained high activity in 15% PKM media and demonstrated strong acid tolerance (70.6% survival at pH 2.5), bile salt tolerance (62.84% at 0.3%), and thermal resistance (83.15% at 42°C). It showed 83.75% hydrophobicity, 73.32%–71.64% autoaggregation, and 78.13% coaggregation, along with marked inhibition against Escherichia coli, Salmonella Enteritidis, and Staphylococcus aureus (15.07–17.12 mm inhibition zones). Natural media composed of 70% coconut water + 30% shrimp wastewater supported optimal enzymatic performance. The B. subtilis–L. fermentum consortium demonstrates potent synergistic enzymatic and probiotic traits, indicating its suitability as a bioenhancer for PKM-based poultry feed. This dual-function probiotic could lower feed costs, improve nutrient digestibility, and support sustainable poultry production. Future work should validate these results through in vivo trials and large-scale fermentation optimization.

Getah virus (GETV), a mosquito-borne alphavirus of veterinary importance, has caused periodic outbreaks in domestic animals, especially in Asia. Although several studies have reported evidence of infection in animals, the overall global seroprevalence remains unclear. This study aimed to comprehensively synthesize available evidence on the worldwide seroprevalence of GETV in domestic and wild animals and identify epidemiological patterns across host types, regions, and detection methods. A systematic search of PubMed, Scopus, ScienceDirect, and Web of Science was conducted following Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 guidelines, without temporal or regional restrictions. Eligible studies reporting serological detection of GETV antibodies in animal populations were included. Data were extracted and analyzed using a DerSimonian–Laird random-effects model, with subgroup analyses stratified by country, host category, sampling period, and diagnostic method. Heterogeneity was quantified using I2 statistics, and potential publication bias was assessed with funnel plots and Egger’s regression test. Fifteen studies (n = 10,211 animals) met the inclusion criteria. The pooled global seroprevalence of GETV was 33.3% (95% confidence interval: 24.2–43.9; I2 = 98.65%, p < 0.001). Malaysia reported the highest seroprevalence (77.2%), followed by China (41.8%) and South Korea (26.4%). Domestic animals (34.0%) exhibited higher exposure than wild species (29.2%), with pigs (43.1%) and cattle (43.2%) recording the highest rates. Studies using virus-neutralization tests yielded higher estimates (47.3%) than those employing enzyme-linked immunosorbent assay (31.4%) or hemagglutination inhibition (7.3%). Meta-regression revealed study location and diagnostic method as significant sources of heterogeneity. This meta-analysis demonstrates that GETV is endemic among Asian animal populations, particularly domestic livestock, indicating substantial virus circulation across species. The findings underscore the need for enhanced veterinary surveillance, standardized serological testing, and One Health-oriented monitoring frameworks to detect and mitigate GETV transmission risks. The absence of data from Africa, Europe, and the Americas highlights an urgent need for geographically expanded research to better understand the virus’s global distribution and zoonotic potential.

Antimicrobial resistance (AMR) resulting from antibiotic misuse in livestock poses a growing threat to animal and human health. The development of sustainable probiotic alternatives supports the United Nations Sustainable Development Goals (SDGs) for zero hunger (SDG 2), good health and well-being (SDG 3), and responsible consumption and production (SDG 12). This study aimed to isolate, characterize, and perform genomic analysis of lactic acid bacteria (LAB) from swine feces exhibiting antibacterial activity against pathogenic Escherichia coli, to explore their potential as eco-friendly probiotic feed additives. Thirty fecal samples were collected from slaughtered crossbred pigs in Thailand. LAB isolates were screened for antibacterial activity against five E. coli pathotypes (Enteroaggregative E. coli, enterohemorrhagic E. coli, enteroinvasive E. coli, enterotoxigenic E. coli, and enteropathogenic E. coli) and assessed for acid and bile tolerance, adhesion capacity, and gastrointestinal survival. Two promising isolates (ATP111 and ATP210) were subjected to whole-genome sequencing and bioinformatics analyses for genes related to antimicrobial production, stress tolerance, virulence, and AMR. Among 93 initial isolates, Lactiplantibacillus argentoratensis ATP111 and Weissella cibaria ATP210 exhibited broad-spectrum inhibition against all E. coli pathotypes. Both strains survived under pH 2.5 and 1% bile conditions, showing 74.39% and 66.90% survival, respectively, in simulated gastrointestinal conditions. Genomic analyses revealed the presence of genes encoding bacteriocins, polyketide synthases, terpenes, and multiple stress-response proteins, supporting their resilience and antimicrobial functionality. Importantly, both genomes lacked virulence and AMR genes, confirming biosafety for probiotic use. The integrated phenotypic and genomic evidence positions L. argentoratensis ATP111 and W. cibaria ATP210 as safe, effective, and sustainable probiotic candidates for swine health management. Their application as antibiotic alternatives aligns with SDG 3 (good health and well-being), SDG 12 (responsible consumption and production), and SDG 15 (life on land), contributing to reduced antibiotic dependence and improved livestock sustainability. Future in vivo validation is recommended to confirm efficacy and support global AMR mitigation efforts.

Growth retardation syndrome in cultured Penaeus vannamei has been associated with Enterocytozoon hepatopenaei (EHP) and a recently identified decapod hepanhamaparvovirus (DHPV) genotype V. However, data on its prevalence, pathogenicity, and interaction with the shrimp hepatopancreatic microbiome in Thailand remain limited. This study aimed to determine the incidence and co-infection rate of DHPV genotype V with EHP, evaluate its pathogenic potential, and explore microbiome alterations associated with infection. Between 2022 and 2023, 1,270 shrimp from 127 grow-out ponds across 46 farms in eastern Thailand and post-larvae 12 from five hatcheries in the south were screened for DHPV and EHP by polymerase chain reaction. Six representative isolates underwent phylogenetic analysis based on non-structural protein 1 (NS1) and NS2 genes. Pathogenicity was evaluated by immersion challenge bioassays in specific pathogen-free P. vannamei. Hepatopancreatic microbiomes of naturally infected and healthy shrimp were compared using 16S ribosomal RNA gene sequencing and Quantitative Insights Into Microbial Ecology 2-based analysis. DHPV was detected in 54.33% (69/127) of ponds and 4% (1/25) of hatchery tanks. Co-infection with EHP occurred in 40.16% of ponds. Phylogenetic analysis showed 97.99%–98.82% similarity with DHPV genotype V from South Korea, confirming transboundary genetic relatedness. Experimental infection caused low mortality (20%) but resulted in viral replication (101–103 copies/μL) and characteristic intranuclear inclusion bodies in hepatopancreatic cells. DHPV-infected shrimp exhibited distinct microbiome profiles with elevated Firmicutes, Planctomycetota, and Actinobacteriota abundances, supporting a pathobiome shift during infection. This is the first report of DHPV genotype V in P. vannamei from Thailand and its frequent co-infection with EHP. Despite its low experimental virulence, the widespread occurrence and microbiome dysbiosis suggest that it may have subclinical impacts that could exacerbate growth retardation. Routine molecular screening in hatcheries and farms, coupled with integrated viral–microbiome surveillance, is essential for sustainable aquaculture biosecurity and aligns with the United Nations Sustainable Development Goal 14 (Life Below Water) by promoting resilient aquatic food systems.

Rabies remains a fatal but preventable zoonotic disease causing nearly 59,000 human deaths annually worldwide, including approximately 75 cases/year in Vietnam. Despite the National Rabies Prevention and Control Program (2022–2030) targeting zero human deaths, community-level awareness and responsible dog management remain key determinants of program success. This study assessed knowledge, attitudes, and practices related to rabies prevention and dog management among residents of Hung Yen province. A community-based cross-sectional survey was conducted among 210 residents from Khoai Chau, Van Lam, and Yen My districts between November and December 2022. Data were collected using a structured, pre-tested questionnaire covering demographics, rabies knowledge, and dog ownership practices. Descriptive statistics and Chi-square tests were performed in R software, with p < 0.05 considered statistically significant. Among 210 respondents, 82.4% had heard of rabies, and 60.7% demonstrated good knowledge. Most knew that dog bites transmit rabies (97.7%), but 39.9% were unaware that scratches can also cause infection. More than half (52.6%) still believed traditional remedies could prevent rabies, and 39.7% did not seek medical care after dog bites. Younger respondents (18–40 years) had significantly better knowledge (odds ratio = 1.90; p = 0.04). Among 85 dog owners, 82.4% vaccinated their dogs, yet 21.2% allowed dogs to roam freely, and 91.8% did not spay or castrate them. Higher socioeconomic status and urban residence were associated with better dog management practices. Despite Hung Yen’s low rabies fatality, notable gaps persist in community awareness and responsible pet ownership. Misconceptions about transmission routes and reliance on traditional treatment threaten progress toward the 2030 rabies-free goal. Strengthened One Health-based education, targeting older adults and low-income rural groups, is recommended to promote timely post-exposure prophylaxis and sustainable dog vaccination and population control programs.

Cryopreservation is an essential assisted reproductive technology for preserving valuable animal genetics; however, it induces osmotic and oxidative stress that compromises sperm quality. Melatonin (MLT) functions as both an antioxidant and a signaling molecule through specific membrane receptors, melatonin 1 (MT1) and melatonin 2 (MT2). This study aimed to determine the association between sperm freezability, MLT receptor expression on spermatozoa, and MLT concentration in the seminal plasma of goat bucks. Semen samples were collected weekly from seven clinically healthy Saanen bucks (2–3 years) maintained under natural photoperiod and standardized feeding conditions. A total of 124 ejaculates were analyzed seasonally (spring-winter). Sperm quality parameters, including motility, viability, capacitation status (chlortetracycline [CTC] assay), plasma membrane fluidity, and acrosome integrity, were evaluated before and after cryopreservation. Expression levels of MT1 and MT2 receptors were quantified by Western blot, while MLT concentration in seminal plasma was measured by enzyme-linked immunosorbent assay. Pearson’s correlation and determination coefficients (R, R2) were computed to assess associations between sperm quality, receptor expression, and MLT concentration. Western blot analysis revealed variable MT1 (16 kDa) and MT2 (28–75 kDa) expression throughout the year. High negative correlations (R > −0.9, R2 > 0.8) were observed between the expression of both receptors and the proportion of acrosome-reacted (AR) spermatozoa (CTC-AR pattern), except for MT2 (75 kDa). Conversely, seminal plasma MLT concentration showed a strong positive correlation (R > 0.9, R2 > 0.8) with capacitated sperm having intact acrosomes (CTC-B pattern). Seasonal variation in receptor expression, rather than MLT concentration, influenced sperm cryoresistance. Sperm freezability in goats is closely linked to the expression of MLT receptors and seminal plasma MLT concentration. Higher receptor expression corresponds to improved post-thaw sperm quality, likely by mitigating cryocapacitation and acrosomal damage. These findings suggest that enhancing MLT receptor expression or modulating photoperiodic exposure could optimize semen cryopreservation protocols and reproductive efficiency in caprine species.

Proteases are key enzymes that hydrolyze peptide bonds enhancing the utilization of feed protein, improving nutrient efficiency, and reducing the need for costly protein ingredients. Despite their growing use in animal nutrition, comparative studies between fungal and microbial proteases in ruminants remain scarce. This study aimed to evaluate the influence of fungal and microbial proteases (25 U/g each) on the microbial composition of rumen fluid and feces, and on the chemical composition of digestive contents in Kazakh White Head bulls. Twenty bulls (14–15 months old; 310–320 kg) fitted with rumen fistulas were divided into three groups: a control group and two treatment groups, each receiving a basal diet supplemented with either fungal or microbial protease. Rumen fluid and fecal samples were analyzed for taxonomic profiles using next-generation sequencing (MiSeq, Illumina) of the 16S ribosomal RNA V3–V4 region. Chemical composition (dry matter [DM], crude protein [CP], crude fat [CF], crude fiber, and ash) was determined according to GOST mass fraction of DM 31640, mass fraction of CP 13496.4, mass fraction of CF 13496.15, mass fraction of crude fiber 31675, and mass fraction of crude ash 26226 standards. Statistical analysis was performed using the Mann–Whitney U-test (p ≤ 0.05). Microbial protease supplementation increased the abundance of beneficial phyla Bacillota (70.1%) and Bacteroidota (19.5%) in rumen fluid, with a corresponding rise in DM (+6.3%), CF (+9.4%), and CP (+7.9%) relative to control. In feces, Bacillota (70.7%) and Bacteroidota (15.5%) predominated. No opportunistic pathogens (e.g., Pseudomonas and Sutterella) were detected in the microbial protease group, indicating improved microbial balance and intestinal protection. Fungal protease exerted milder effects, with modest increases in nutrient fractions. Microbial protease was more effective than fungal protease in optimizing rumen microbiota and enhancing nutrient digestibility in bulls. Its use may support environmentally sustainable livestock production by reducing nitrogen excretion and dependence on high-protein feed ingredients. These findings provide a scientific basis for breed-adapted enzymatic feeding strategies in ruminants.

Insulin-like growth factor-1 (IGF-1) plays a crucial role in folliculogenesis and oocyte maturation by regulating oxidative stress and apoptotic pathways. However, the optimal IGF-1 concentration for small ruminant oocytes, particularly the native Kacang goat, remains undefined. This study aimed to determine the optimal IGF-1 dose for improving oocyte quality during in vitro oocyte maturation (IVM) by evaluating oxidative stress and apoptosis markers. Ovaries (n = 120) were collected from local slaughterhouses, and cumulus–oocyte complexes were aspirated, selected, and randomly divided into four groups: Control (0 ng/mL IGF-1) and treatments with 50, 100, and 150 ng/mL IGF-1 supplementation. Mature oocytes were analyzed for oxidative stress biomarkers, including superoxide dismutase (SOD-1), glutathione (GSH), and malondialdehyde (MDA), using an enzyme-linked immunosorbent assay, and for apoptosis regulators, B-cell lymphoma 2-associated X protein (BAX), B-cell lymphoma-2 (BCL-2), and cytochrome c, using immunocytochemistry. Data were analyzed using one-way analysis of variance and Duncan’s post hoc test (p < 0.05). IGF-1 supplementation produced concentration-dependent effects. The 100 ng/mL group (T2) exhibited the highest SOD-1 (2.07 ± 0.60) and GSH (8.07 ± 1.79) levels (p < 0.05), while MDA increased with higher IGF-1 doses, indicating a threshold beyond which oxidative stress is induced. Anti-apoptotic BCL-2 and cytochrome c expressions peaked at 50 ng/mL IGF-1 (10.73 ± 1.56 and 11.73 ± 0.99, respectively), whereas the pro-apoptotic marker BAX was lowest in the same group and increased at higher doses. The findings suggest that 50 ng/mL IGF-1 effectively maintains redox balance and mitochondrial stability through phosphatidylinositol 3-kinase/protein kinase B-mediated regulation. This study demonstrates, for the 1st time, a dose-dependent, biphasic effect of IGF-1 on oxidative and apoptotic pathways in Kacang goat oocytes. An optimal concentration of 50 ng/mL IGF-1 enhances oocyte maturation by balancing antioxidant defense and anti-apoptotic mechanisms, whereas higher concentrations induce oxidative stress. These findings establish a breed-specific IVM optimization protocol that supports genetic preservation and sustainable reproductive biotechnology for indigenous goats.

Reproductive efficiency during the non-breeding season is a critical factor influencing year-round productivity in sheep farming, particularly in semi-arid environments. Pregnant mare serum gonadotropin (PMSG) is commonly used to induce estrus, yet the optimal dosage that maximizes fertility and lamb growth while minimizing hormonal side effects remains unclear. This study aimed to determine the optimal PMSG dose for enhancing reproductive and growth performance in Dorper × Assaf crossbred ewes under semi-extensive conditions in Palestine. A total of 143 non-lactating Dorper × Assaf ewes (aged 2–3 years) were synchronized with intravaginal progestagen sponges for 14 days, followed by intramuscular administration of 400, 500, or 600 international units (IU) of PMSG. Key reproductive traits, estrus response, conception rate, abortion rate, fecundity, and lamb survival, were recorded, alongside lamb birth and weaning weights. Data were analyzed using a one-way analysis of variance and a general linear model incorporating parity, litter size, and sex effects, with significance set at p < 0.05. High conception and lambing rates were observed across all groups. The 400 IU PMSG dose resulted in the lowest abortion rate (2.44%) and highest lamb survival (96.9%), while higher doses (500–600 IU) increased abortion incidence without improving fertility outcomes. Birth weight increased with PMSG level (3.42–3.83 kg), whereas weaning weight peaked at 500 IU (22.18 kg). Litter size and lamb sex significantly affected both birth and weaning weights, with singletons and males being heavier. Administering 400 IU of PMSG provides the optimal balance between reproductive performance, lamb survival, cost-efficiency, and animal welfare. Excessive hormonal stimulation at higher doses offers no additional reproductive benefit and may compromise ewe health. Implementing this moderate, breed-specific hormonal protocol enhances fertility while reducing veterinary intervention and production costs, promoting sustainability and welfare-conscious management. These outcomes directly contribute to sustainable development goals (SDG) 2 – zero hunger and SDG 12 – responsible consumption and production, advancing resilient and ethical small-ruminant farming systems.

Indigofera zollingeriana is a high-protein tropical legume with potential as a sustainable ruminant feed; however, its ensiling is challenged by rapid proteolysis and ammonia accumulation. Incorporating lactic acid bacteria (LAB) inoculants and natural tannin sources may enhance fermentation quality and nitrogen preservation. This study evaluated the effects of Lactiplantibacillus plantarum inoculant and Acacia mangium leaf tannin extract, individually and in combination, on the chemical composition, ensiling characteristics, in vitro rumen fermentation, digestibility, and metabolomic profiles of Indigofera silage. A completely randomized design was used with four treatments: (R0) control, (R1) L. plantarum (10⁶ colony forming units/g FM), (R2) 1% A. mangium extract, and (R3) combination of both additives, each with five replicates. Silages were fermented anaerobically for 30 days. Analyses included proximate composition, pH, lactic acid, ammonia nitrogen (NH3-N), in vitro gas and methane (CH4) production, volatile fatty acids (VFA), digestibility, and untargeted metabolomics of both silage and rumen fluid using gas chromatography–mass spectrometry. Data were evaluated using a one-way analysis of variance, Duncan’s test, principal component analysis, and partial least squares discriminant analysis. L. plantarum lowered (p < 0.05) silage pH and fiber fractions but increased NH3-N due to enhanced deamination. A. mangium tannins effectively suppressed proteolysis, reducing NH3-N by 11.85%. Their combination improved (p < 0.05) dry- and organic-matter digestibility (↑ ≈ 9%), increased the propionate proportion by 6.82%, and lowered the acetate-to-propionate ratio, indicating a shift toward more energy-efficient rumen fermentation without significant methane (CH4) inhibition. Metabolomic profiling identified 23 key metabolites in silage and 11 in rumen fluid; acacia tannins increased fatty acyl compounds (+14.3%) while LAB enhanced prenol lipids, reflecting modified lipid and oxygen-derived pathways that improve nutrient utilization. The combination of L. plantarum and A. mangium leaf extract synergistically improved Indigofera silage quality, nutrient preservation, and digestibility while modulating beneficial metabolites associated with rumen fermentation efficiency. This integrated additive strategy represents a sustainable and locally adaptable approach for tropical ruminant feed production.

Milk yield variability in tropical dairy goats is driven not only by nutrition but also by complex metabolic and hormonal regulation. Conventional nutrition studies often overlook the physiological mechanisms underlying lactation efficiency. This study aimed to integrate metabolomic and hormonal analyses to identify biomarkers associated with high and low milk yield performance in Sapera goats. It provides the first untargeted ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (UHPLC-HRMS)-based metabolomics characterization linking metabolic and endocrine profiles to lactational efficiency in tropical dairy systems. Twenty lactating Sapera goats were categorized into two groups: High-yielding (HY) (>1000 mL/day) and low-yielding (LY) (≤1000 mL/day). All animals were fed identical diets formulated according to National Research Council (2007) standards and managed under uniform housing conditions. Milk composition, blood metabolites (glucose, cholesterol, total protein, and triglycerides), and plasma hormones (leptin and insulin-like growth factor-1) were quantified using colorimetric and enzyme-linked immunosorbent assay assays. Untargeted metabolomics of milk samples was performed using UHPLC-HRMS. Metabolites were identified through multi-database annotation, and statistical comparisons were conducted using independent t-tests with p < 0.05. Milk composition did not differ significantly between groups (p > 0.05). However, HY goats exhibited higher glucose and cholesterol concentrations and lower leptin levels (2.39 ± 0.42 ng/mL vs. 3.00 ± 0.44 ng/mL). Metabolomic analysis identified 213 metabolites, 19 unique to HYs, 28 to LYs, and 166 metabolites were found in both groups. HY goats showed enrichment of metabolites linked to lipid metabolism, membrane synthesis, and antioxidant defense (e.g., uric acid and phosphoserine derivatives), while LY goats displayed compounds indicative of metabolic stress and detoxification load (e.g., glycocholic acid and 3-furoic acid). Integrative correlation mapping revealed coordinated regulation between blood and milk metabolites in HY animals. HY and LY goats possess distinct metabolic and hormonal signatures despite similar milk composition. Identified biomarkers such as uric acid and glycerophosphoserine highlight pathways supporting efficient nutrient utilization and milk synthesis. These findings provide a foundation for precision feeding and biomarker-guided selection strategies to enhance productivity and sustainability in tropical dairy goat systems.

Vitrification is a promising cryopreservation technique for conserving the genetic resources of Aceh cattle; however, it may induce cryoinjury and follicular apoptosis. Pituitary extract (PE), containing follicle-stimulating hormone (FSH) and luteinizing hormone (LH), can potentially mitigate apoptosis and maintain follicular viability. This study investigated the effect of bovine PE supplementation in vitrification media on the expression of caspase-3, FSH receptor (FSHR), and LH receptor (LHR) in vitrified ovarian tissue of Aceh cattle. A factorial, completely randomized design was employed using five vitrification media, PE at 0, 200, 400, and 600 μg/mL, and commercial bovine PE (Gibco, 300 μg/mL), combined with three vitrification durations (0, 7, and 14 days), each in triplicate. Post-vitrification ovarian tissue was evaluated by immunohistochemistry for caspase-3, FSHR, and LHR expression. The intensity scores (IS) were analyzed using the Kruskal–Wallis and Mann–Whitney U tests (p < 0.05). PE supplementation significantly (p < 0.05) reduced caspase-3 expression across all follicular stages, indicating decreased apoptosis, whereas vitrification duration alone showed limited effects. The lowest caspase-3 IS (negative) occurred in the 600 μg/mL PE group after 7 days. Both PE concentration and vitrification duration significantly influenced FSHR and LHR expression (p < 0.05), with strong positive interactions between the two factors. FSHR and LHR expression increased progressively from primordial to antral follicles, suggesting preserved gonadotropin responsiveness. Optimal follicular preservation and receptor integrity were achieved at 600 μg/mL PE following 7 days of vitrification. Supplementation of vitrification media with 600 μg/mL bovine PE effectively suppresses follicular apoptosis while enhancing FSHR and LHR expression, maintaining ovarian functionality after vitrification. These results highlight PE as a low-cost, multi-hormonal alternative to single-gonadotropin supplements in cryopreservation media, supporting sustainable genetic conservation of Aceh cattle. Integrating locally derived PE into cryopreservation protocols could improve post-thaw follicular survival, reduce dependency on commercial reagents, and strengthen reproductive biotechnology for indigenous livestock conservation.

The giant freshwater prawn (Macrobrachium rosenbergii) is an economically valuable aquaculture species, yet its production often faces challenges related to poor growth and disease outbreaks caused by intensive farming practices and excessive antibiotic use. Probiotics offer a sustainable alternative for improving growth, immunity, and pond health, but their efficacy and safety are highly strain specific. This study aimed to evaluate the probiotic potential of Bacillus pseudomycoides, Bacillus safensis, and Bacillus infantis isolated from prawn farms in Kalasin Province, Thailand, focusing on digestive enzyme activities, hemolytic safety, and antibiotic susceptibility. Three Bacillus strains previously identified by 16S ribosomal RNA sequencing were characterized using standard in vitro assays. Amylase, protease, and lipase activities were assessed using the halo/colony (H/C) ratio and enzyme unit measurements. Hemolytic patterns were examined on Columbia agar supplemented with 5% sheep blood, and antibiotic susceptibility was tested by the disk diffusion method against seven antibiotics following Clinical and Laboratory Standards Institute (2024) guidelines. All assays were performed in triplicate, and data were analyzed using one-way analysis of variance (p < 0.05). All strains exhibited strong amylolytic (H/C ratio 2.05–2.27) and proteolytic (H/C ratio 1.75–1.96) activities, while lipase activity was undetectable. Hemolysis testing revealed γ-hemolysis for all strains, confirming non-hemolytic and non-pathogenic properties. Antibiotic susceptibility profiles indicated broad sensitivity to penicillin G, tetracycline, streptomycin, erythromycin, and chloramphenicol. Moderate susceptibility to vancomycin was observed in B. pseudomycoides and B. safensis, while B. infantis remained fully susceptible. The results suggest a strong digestive enzyme potential and an acceptable safety profile among all isolates. The evaluated Bacillus strains exhibit favorable probiotic attributes, including high amylase and protease activity, non-hemolytic safety, and broad antibiotic susceptibility, which support their suitability for probiotic application in M. rosenbergii culture. These native isolates may serve as sustainable, locally adapted alternatives to antibiotics, contributing to improved feed efficiency, growth, and disease resistance in freshwater prawn aquaculture. Further in vivo validation and genomic safety analyses are recommended to confirm their efficacy under commercial conditions.

Sperm cryopreservation is a cornerstone technology for genetic resource conservation and artificial insemination. However, tropical climatic conditions often compromise post-thaw sperm quality due to enhanced oxidative and thermal stress. This study aimed to optimize glycerol and egg yolk concentrations in a Tris–citrate–glucose (TCG) extender to improve post-thaw sperm quality of Saanen bucks raised in the Mekong Delta, Vietnam. The experiment was conducted in two sequential phases. In Phase 1, five glycerol concentrations (4%, 6%, 8%, 10%, and 12%) were tested to identify the optimal permeating cryoprotectant level. In Phase 2, four egg yolk concentrations (5%, 10%, 15%, and 20%) were assessed in combination with the optimal glycerol concentration. Semen from four healthy Saanen bucks was collected using an artificial vagina, diluted in the respective extenders, equilibrated, and frozen in liquid nitrogen. Post-thaw evaluations included overall and progressive motility (phase-contrast microscopy), viability (eosin–nigrosin staining), membrane integrity (hypo-osmotic swelling test), acrosome integrity (Giemsa staining), and DNA fragmentation (sperm chromatin dispersion test). Data were analyzed using one-way analysis of variance with Tukey’s post hoc test at p < 0.05. Glycerol concentration had a significant effect on post-thaw sperm quality (eta squared = 0.93–0.97), with 10% providing the best balance between cryoprotection and cytotoxicity. Incorporating 15% egg yolk with 10% glycerol significantly improved sperm viability (65.5%), membrane integrity (72.3%), and reduced DNA fragmentation (12.4%) compared with other treatments (p < 0.05). Higher glycerol or egg yolk concentrations adversely affected motility due to increased osmotic stress and viscosity. The combination of 10% glycerol and 15% egg yolk in TCG extender provides optimal cryoprotection for Saanen buck semen under tropical conditions. The resulting post-thaw sperm exhibited high motility, viability, and DNA integrity. This protocol can serve as a region-specific standard for buck semen cryobanking and artificial insemination in tropical climates, supporting genetic improvement and conservation initiatives in Vietnam and other developing regions.

The emergence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in food sources poses a growing threat to public health. Poultry eggs may act as vehicles for these resistant bacteria, facilitating their transmission through the food chain. This study aimed to determine the prevalence, antimicrobial resistance (AMR) patterns, and ESBL-encoding genes among E. coli isolates recovered from chicken, duck, and quail eggs sold in Can Tho City, Vietnam. A total of 900 eggs (300 each from chicken, duck, and quail) were collected from local markets, retail stores, and supermarkets between June and December 2024. E. coli isolates were identified through biochemical tests and polymerase chain reaction (PCR) targeting the uidA gene. ESBL production was determined using the disk diffusion method, and the presence of blaCTX-M, blaTEM, blaNDM, and blaSHV genes was confirmed by PCR. Antimicrobial susceptibility was tested against 10 antibiotics representing eight classes following Clinical and Laboratory Standards Institute (2021) guidelines. Out of 179 pooled samples positive for E. coli, 52 isolates (29.1%) were confirmed as ESBL producers. The highest prevalence was observed in chicken (32.4%) and duck (32.8%) eggs, while quail eggs showed a prevalence of 20%. ESBL-producing isolates most frequently carried blaCTX-M (65.4%) and blaTEM (44.2%) genes, whereas blaNDM was detected in 1.9% of isolates and blaSHV was absent. All isolates were multidrug-resistant, exhibiting resistance to 4–9 antibiotic classes. High resistance was observed to amoxicillin-clavulanate (69.2%), ceftriaxone (69.2%), tetracycline (75%), and trimethoprim-sulfamethoxazole (61.5%). No E. coli was detected in supermarket eggs, suggesting improved hygiene practices reduce contamination. The detection of ESBL-producing and multidrug-resistant E. coli on poultry eggshells underscores a significant public health concern. The predominance of blaCTX-M and blaTEM genes highlights the risk of resistance gene dissemination through the egg supply chain. Enhanced surveillance, responsible antibiotic use, and strict hygiene control in small-scale poultry production systems are urgently needed to mitigate the spread of AMR under the One Health framework.

Hepatocystis, a malaria-like hemoparasite closely related to Plasmodium, infects non-human primates (NHPs), bats, and other mammals, yet remains understudied in Indonesia. Although Plasmodium detection in primates has been extensively reported, molecular confirmation of Hepatocystis in Indonesian wildlife is lacking. This study aimed to screen NHP fecal samples for Plasmodium infection and to identify any malaria-like parasites using molecular methods. A total of 227 fecal samples from captive and rescued NHPs, representing multiple Macaca species and other primates, were collected from Tasikoki Wildlife Rescue Center, Manado, Indonesia, in 2019 and 2021. Genomic DNA was extracted using a QIAamp Fast DNA Stool Mini Kit (Qiagen, Germany) and screened for Plasmodium using polymerase chain reaction (PCR) targeting the mitochondrial small subunit ribosomal RNA gene. Positive amplicons were purified, sequenced, and analyzed using the basic local alignment search tool and phylogenetic reconstruction with MEGA X. Eight (3.5%) of 227 samples yielded positive PCR bands of approximately 600 bp, differing from the expected 467 bp for Plasmodium. Sequencing of four representative samples (MNig-01, MNig-17, MNig-18, and HM-160) revealed >99.7% identity with Hepatocystis spp. (GenBank: KY653782.1). Multiple sequence alignment confirmed complete nucleotide conservation among the four isolates, and phylogenetic analysis clustered them within the Hepatocystis clade, closely related to Hepatocystis spp. ex Pteropus hypomelanus from Malaysia. All positive detections were from 2019 samples, suggesting temporal variation in infection or vector activity. This study provides the first molecular evidence of Hepatocystis infection in Indonesian NHPs using fecal DNA, expanding current knowledge of parasite distribution and host range. The successful detection of Hepatocystis through non-invasive sampling underscores the potential of fecal-based PCR for wildlife disease surveillance. These findings highlight the importance of integrating molecular diagnostics into conservation and One Health frameworks to monitor zoonotic parasites and understand host–vector–pathogen interactions in natural ecosystems.