Search Results (11,036)

African swine fever virus (ASFV) antibody detection is an important component of serological surveillance and retrospective investigations. DP238L was recently identified by proteome-wide serum-antibody profiling as a frequently recognized ASFV antigen, and recombinant DP238L was further shown to react with ASFV-positive sera and to elicit antigen-specific antibody responses in immunized pigs. Here, we extend those observations toward diagnostic assay development and preliminary validation. Full-length DP238L from ASFV strain SY-1 (GenBank OM161110) was evaluated as a single recombinant coating antigen for an indirect ELISA (DP238L-iELISA). Independent sequence analysis showed that SY-1 DP238L contains 238 amino acids, with a theoretical molecular mass of 27.13 kDa, a theoretical pI of 4.92, and a GRAVY value of −0.97. In a quality-restricted subset of complete, unambiguous 238-aa homologs (n = 7), amino acid identity to SY-1 ranged from 97.48% to 100% (median, 98.74%). Recombinant DP238L was expressed and purified from Escherichia coli and specifically recognized by ASFV-positive swine serum. ELISA optimization identified a coating antigen concentration of 0.5 μg/mL, a secondary-antibody working dilution of 1:2500 (0.4 μg/mL), a serum dilution of 1:100, a serum incubation time of 45 min, and a substrate development time of 15 min. Using 518 clinical swine serum samples categorized according to the commercial INgezim PPA COMPAC K3 blocking ELISA, ROC analysis yielded an AUC of 0.9989 (bootstrap 95% CI, 0.9973–0.9999). At the selected S/P cutoff of 0.30, positive percent agreement and negative percent agreement relative to INgezim were 97.2% and 98.7%, respectively; overall agreement was 98.3% and Cohen’s $\kappa$ was 0.956. A limited six-sample heterologous panel provided preliminary cross-reactivity information, with all tested samples remaining below the cutoff. In a longitudinal panel from five experimentally infected pigs, three animals exceeded the cutoff at 7 days post-infection (dpi) and all five exceeded it by 10 dpi. These findings support DP238L as a promising recombinant antigen for ASFV serological assay development while emphasizing that the present results represent an internal comparator-based preliminary evaluation requiring broader independent validation. Full article

Brucellosis is an important zoonotic disease worldwide, and Brucella canis (B. canis) is an emerging concern in companion animal health. Rapid and reliable detection of B. canis in dogs is essential for disease control. Herein, we developed a one-pot MIRA CRISPR/Cas12b fluorescence biosensing platform targeting the conserved omp31 gene. The platform integrates MIRA mediated isothermal amplification with CRISPR/Cas12b-based target recognition and collateral cleavage. After optimization, the assay achieved a detection limit of 5 copies/reaction within 20 min, comparable to qPCR. No cross reactivity was observed with seven common canine pathogens. Among 128 clinical blood samples, the biosensor showed complete agreement with qPCR, achieving 100% sensitivity and 100% specificity. These findings demonstrate that the biosensor provides a rapid, sensitive, and specific approach for B. canis detection, with potential for decentralized veterinary testing through its endpoint visual fluorescence readout and applications in One Health surveillance. Full article

Background: The gut–vascular axis linking gut microbiota, intestinal barrier, and vascular endothelium is a key pathophysiological interface in chronic metabolic diseases. Dysbiosis and “leaky gut” enable bacterial products and gut-derived metabolites to enter the circulation, driving systemic inflammation and endothelial dysfunction. Single-target interventions show limited efficacy; combined gut-directed and endothelium-directed strategies remain underexplored. Methods: PubMed was searched from database inception to 31 August 2026 using a predefined search strategy combining terms for the gut–vascular axis, intestinal barrier, microbiota/metabolites, endothelial dysfunction, and combined interventions. Preclinical and clinical studies evaluating gut barrier integrity, microbiota modulation, or endothelial protection, alone or in combination, were included. No formal risk-of-bias scoring was performed; study design, sample size, endpoint type, follow-up, and limitations were appraised. Results: (1) The axis comprises three core components: gut barrier, endothelium, and microbial metabolites. (2) Pathological activation involves leaky-gut-driven endotoxemia, dysbiosis-induced metabolic inflammation, and immune crosstalk sustaining vascular inflammation. (3) Animal models demonstrate that combined interventions can reduce atherosclerotic plaque, improve endothelium-dependent relaxation, and lower systemic inflammation to a greater extent than some monotherapies; however, most studies have not directly compared combination therapy with both corresponding monotherapies or performed formal interaction testing, so true synergy remains unproven. (4) Limited clinical studies in metabolic syndrome and heart failure suggest improvements in surrogate endpoints, but large-scale randomised trials with hard cardiovascular endpoints are lacking. Conclusions: The gut–vascular axis is a promising therapeutic framework for cardiometabolic diseases, with a strong biological rationale. Combined gut- and endothelium-directed interventions offer a rational multi-target approach, but direct evidence that they provide superior clinical outcomes remains limited. Future research must prioritise hard-endpoint trials, standardised interventions and endpoints, long-term safety, reproducibility, multi-omics-based patient stratification, and next-generation microbiome therapies. Full article

Against the dual goals of agricultural and rural carbon emission reduction, carbon sequestration and food security guarantee, this paper uses municipal panel data of 21 prefectures in Sichuan Province from 2015 to 2024 to calculate agricultural carbon emissions from multiple dimensions including farmland utilization, paddy field methane, crop cultivation, straw burning and livestock breeding. The standard deviational ellipse model (SDE) is adopted to reveal the spatiotemporal pattern of agricultural carbon emissions, and the Tapio decoupling model is applied to identify the decoupling relationship between agricultural carbon emissions and the agricultural economy. A panel model is constructed. The Breusch–Pagan Lagrange Multipkier (LM) test and Hausman test are performed to select the random-effects model to examine the impact of agricultural technological progress on agricultural carbon emissions. Robustness tests and grouped heterogeneity analysis are carried out. Furthermore, mediation and moderation models are introduced to identify the internal transmission paths and boundary conditions. The results show that: (1) From 2015 to 2024, the total agricultural carbon emissions in Sichuan Province generally declined. Animal husbandry was the primary emission source while carbon emissions from crop cultivation showed an upward trend, and carbon emission intensity kept decreasing. The spatial pattern of agricultural carbon emissions presented strong path dependence; the emission centroid remained stable in the central basin with a northeast–southwest spatial distribution. (2) The relationship between agricultural carbon emissions and the agricultural economy in Sichuan Province fluctuated alternately between strong decoupling and weak decoupling. Sustained and stable strong decoupling has not yet been achieved, and the incremental emissions from crop cultivation constitute a key factor disturbing the decoupling state. (3) Benchmark regression results indicated that agricultural technological progress significantly curbed regional agricultural carbon emissions, and urbanization also exerted emission-reduction effects. The proportion of primary industry, land scale operation and grain planting ratio significantly increased agricultural carbon emissions, whereas rural residents’ income and agricultural industrial structure showed no significant impacts. The carbon emission reduction effect of agricultural technological progress, constrained by regional endowments, presented notable regional heterogeneity. The emission reduction effect of technological progress was significant in plain and non-ethnic farming areas, while it was difficult to achieve emission reduction merely by relying on mechanization in ethnic regions and non-plain mountainous areas. (4) Mechanism tests demonstrated that fertilizer application intensity played a partial mediating role between agricultural technological progress and agricultural carbon emissions. Agricultural technological progress can indirectly reduce carbon emissions by lowering fertilizer application intensity. The multiple cropping index exerted a positive moderating effect. The higher the multiple cropping index, the stronger the inhibitory effect of agricultural technological progress on agricultural carbon emissions. This paper identified key regions, core emission reduction targets and directions for technological optimization for differentiated low-carbon agricultural governance in Sichuan Province, and provided empirical evidence for developing agriculture-specific carbon emission reduction plans tailored to local conditions. It also clarified the limits of the emission reduction effectiveness of agricultural technological advancements across different regions. The research findings can provide precise guidance for the green and low-carbon transformation of regional agriculture, assist local governments in addressing weaknesses in agricultural emission reduction efforts and mitigate risks such as rising emissions from crop cultivation and rebound effects from decoupling. Full article

Emerging influenza viruses and mosquito-borne flaviviruses continue to challenge animal and public health, highlighting the need for antivirals that remain effective after infection. This study evaluated the in vitro activity of a 70% ethanolic whole-specimen extract of the marine bivalve Entodesma navicula (ENE) against influenza A/Puerto Rico/8/1934 (H1N1; A/PR8), A/Wisconsin/15/2009 (H3N2), influenza B/Florida/78/2015, Zika virus, and dengue virus serotype 2. Antiviral activity was assessed using time-of-addition analysis, viability-based cytopathic-effect assays, reverse-transcription quantitative PCR, infectious virus titration, immunofluorescence, Western blotting, and flow cytometry. In the A/PR8 model, pre- and co-treatment produced no consistent inhibition. By contrast, post-infection ENE suppressed all tested viruses. For A/PR8, Zika virus, and dengue virus, this activity was supported by reduced viral genome abundance, infectious progeny, and viral antigen expression. In A/PR8-infected A549 cells, ENE reduced phosphorylation of receptor-interacting serine/threonine-protein kinase 3 (RIPK3) and mixed lineage kinase domain-like pseudokinase (MLKL). ENE also reduced the proportion of cells positive for phosphorylated MLKL within Zika- and dengue-virus-infected Vero E6 populations. These findings identify E. navicula as an underexplored source of post-infection antiviral bioactivity associated with attenuated necroptosis-related signaling. Isolation of active constituents and causal validation in primary cells and animal models are needed. Full article

The present article provides genotype and phenotype data of 3279 Kazakh local horses from various regions of Kazakhstan. Genotypic data are 81,974 single nucleotide polymorphisms (SNPs) from genome-wide genotyping of all individuals sampled. All horses were phenotyped and their height at withers, oblique body length, chest circumference, cannon bone circumference and body mass were recorded. Field sampling of data took place at various study sites and biological samples and morphometric measurements were taken from each animal at the same time. The dataset comprises both raw and quality-controlled genotypic data, as well as phenotypic data in structured format. All animals were identified and cataloged, with metadata such as sampling location, sex and age if applicable. Following QC filtering, 2704 individuals and 73,377 SNPs were left. The dataset serves as a resource for population genetic studies, genome-wide association studies and genomic selection research, and for studies on genetic architecture of morphological traits. It can also be used for comparative genomics among breeds of horses, genetic diversity studies of central Asian horses and the creation and/or testing of genomic prediction models for body measurement traits. The dataset is freely available and formatted for easy integration with other equine genomic databases. Full article

The Chinhai spiny newt (Echinotriton chinhaiensis) is a rare amphibian species endemic to China. Its specialized habitat and small size make conventional field surveys and population monitoring not only time-consuming and labor-intensive, but also challenging. To address this challenge, we developed a visual detection method for habitat monitoring by integrating environmental DNA (eDNA) analysis with multi-enzyme isothermal rapid amplification (MIRA) and lateral flow dipstick (LFD) technologies. A specific mitochondrial gene fragment was selected as a target. MIRA primers and probes were designed and optimized through comparative sequence analysis with sympatric species, including crucian carp, bullfrogs, and polar fire shrimp. The assay conditions were subsequently refined by optimizing reaction temperature and test strip development time. Results indicate that using aquatic eDNA as a template, the optimized MIRA system incubated at 37 °C for 30 min, followed by a 20 min LFD development, produces clear and interpretable results. The assay demonstrated a DNA detection limit of 5.1 × 10⁻⁴ ng/μL, corresponding to a sensitivity higher than that of conventional PCR. The complete MIRA-LFD procedure can be performed within 50 min, and its identification results are consistent with those from high-throughput sequencing. This established method enables specific, efficient, and rapid detection of the Chinhai spiny newt in aquatic environments. It provides a practical technical tool for habitat surveys of this species, improving efficiency while reducing labor and resource demands. Furthermore, this approach offers a potential reference for habitat studies of other rare animal species. Full article

Antimicrobial resistance (AMR) is a critical health issue affected by human, animal, and environmental factors within a One Health framework. Livestock production, particularly pig farming, contributes to the emergence and spread of resistant bacteria through antimicrobial use and selective pressure. This study characterized the antimicrobial resistance profiles of Escherichia coli and enterococci isolated from piglet faeces in Portugal, with emphasis on antimicrobials classified by the World Health Organization (WHO) according to their importance for human medicine. A total of 340 E. coli and 130 Enterococcus isolates were tested using disk diffusion, and the resistance frequencies were calculated at isolate level. E. coli showed high resistance to commonly used antimicrobials, including amoxicillin–clavulanic acid (67.1%) and ciprofloxacin (43.2%), a highest priority critically important antimicrobial (HPCIA). Resistance was also found to human-only drugs such as piperacillin (67.4%) and, at a lower frequency, imipenem (2.6%), a last-resort carbapenem. Multidrug resistance was detected in 61.2% of isolates. Among enterococci, E. faecium exhibited high resistance to imipenem (64.4%), while E. faecalis and other Enterococcus species showed notable resistance to ciprofloxacin. Overall, resistance crossed all WHO antimicrobial categories. This finding highlights the potential role of pig production in the dissemination of AMR across ecosystems and suggests the need for strengthened One Health surveillance and antimicrobial stewardship efforts. Full article

Background/Objectives: Rapid antimicrobial decision-making in small-animal practice is limited by the turnaround time (≥2 days) of conventional laboratory-based culture and antimicrobial susceptibility testing (AST). Existing point-of-care and in-clinic tests can provide faster results, but their performance often depends on user experience. This study evaluated VetBac, a hybrid workflow in which the test is conducted in the clinic, while experts centrally interpret bacterial growth on plates. Methods: We analyzed 60 interpretable plates from canine and feline skin, ear, and urine infections. In the clinic, samples were inoculated onto chromogenic Mueller–Hinton agar plates, after which antibiotic discs were applied. Plates were incubated for 16–20 h, photographed, and interpreted centrally by an expert. For comparison, samples were submitted in parallel to standard microbiology laboratories. Results: Identification of primary bacterial populations in clinical samples showed high species-level agreement with laboratory results (83.9%), increasing to 100.0% when staphylococci species were grouped. Secondary bacteria agreement (16.7%) was lower due to unrecognizability and overgrowth by dominant bacteria. Overall AST agreement was 93.6% (262/280), with 91.9% sensitivity for detecting resistance and 94.0% specificity for detecting susceptibility. VetBac-susceptible discrepancies occurred in 5/62 laboratory-resistant comparisons (8.1%), and VetBac-resistant discrepancies occurred in 13/218 laboratory-susceptible comparisons (6.0%). Conclusions: In interpretable skin, ear, and urine samples, the VetBac workflow showed high agreement with laboratory results for primary bacterial identification and AST. These findings support the hybrid in-clinic/centralized model as an adjunctive source of earlier antimicrobial decision support, pending laboratory confirmation. However, the study did not estimate the proportion of all clinical samples suitable for direct culture and AST. Full article

Real-time quantitative PCR (qPCR) is a key technique for elucidating the mechanisms underlying environmental stress responses in aquatic animals, and its accuracy relies heavily on stable reference genes for data normalization. In this study, histopathological examination revealed alterations in the gill, hepatopancreas, and muscle tissues of Procambarus clarkii following NH₃ stress. Based on transcriptomic screening and reference genes reported in the literature, 13 candidate reference genes were selected. Their expression levels in 8 different tissues before and after NH₃ stress were detected using qPCR, and their stability was systematically evaluated using multiple algorithms, including geNorm, NormFinder, BestKeeper, Delta CT and RefFinder. The results identified 60S ribosomal protein L13a (RPL13a), ribosomal protein S18 (RPS18) and eukaryotic translation initiation factor 4A (EIF4A) as the three most stable candidates under NH₃ stress, whereas the traditionally used reference genes beta-actin (β-actin) and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) showed relatively low stability under these conditions. geNorm pairwise variation analysis indicated that the inclusion of a third reference gene could further improve normalization. Validation using four NH₃-responsive genes showed that (RPL13a + RPS18) and (RPL13a + RPS18 + EIF4A) yielded similar log₂ fold-change estimates, supporting their use as normalization options under the tested conditions. It is worth noting that RPL13a remains highly stable across four conditions: different embryonic developmental stages, different ovarian developmental stages, tissues from both males and females, and following infection with white spot syndrome virus (WSSV). These findings contribute to the selecting and validating reference genes for qPCR analyses in Procambarus clarkii under the specific experimental conditions evaluated in this study. Full article

Cyanobacteria are microorganisms that can produce natural toxins with significant impacts on human, animal and ecosystem health. Portishead Marine Lake is a small freshwater lake in southwest England frequented by thousands of people each year for recreational activities but was associated periodically with prolific cyanobacterial blooms with potential risk to visitor health. Water samples were tested weekly between January and December 2016 for cyanobacteria and cyanotoxins. Citizen scientists undertook testing for nutrients, dissolved oxygen, pH, temperature and meteorological parameters. Four major blooms of Microcystis spp. were observed in the warmer months between May and September and were associated with high concentrations of microcystins. Complex toxin congener profiles were dominated by MC-LR and MC-RR, with lower contributions from MC-WR and MC-YR. Cyanobacterial cell densities and summed microcystin concentrations ranged from 1600 to 5.6 × 10⁶ cells/mL and 0.1 to 18,069 µg/L, respectively, and correlated significantly. Toxin concentrations were positively correlated with nitrate levels and inversely correlated with pH, although mean pH levels did not fall below 7.0. There was little statistical evidence for influence of weather conditions overall, although significant intra-lake differences in cell densities and toxin concentrations were noted. Intervention measures employed by local authorities included the use of barley straw, hydrological manipulation, debris removal and ultrasound treatment. Whilst no statistical assessment of intervention effectiveness was possible without effective control samples, there was some limited evidence for the temporary reduction in toxin levels after hydrological intervention, but none following installation of ultrasound. Overall, the study utilized LC-MS/MS and microscopy to quantify microcystins and Microcystis spp., respectively, and evidenced risks to lake visitors and the importance of toxin testing in recreational waters and the use of appropriate risk mitigation approaches. Further work incorporating appropriate experimental controls would be needed to determine the effectiveness of hazard mitigation measures for reducing or eliminating human health threats. Full article

Saline attached-growth biofilters serving live crustacean cultures are seldom characterised with the quantitative tools of biological nitrogen-removal engineering. Two saline moving bed biofilm reactors (MBBRs) serving live mud crab (Scylla paramamosain) cultures ran for 65 days at nominal hydraulic retention times of 60 and 120 min, as independent case trajectories, not a replicated comparison. Through the 35 days of live-animal loading, both held total ammoniacal nitrogen removal above 85%, including across reduced-feeding, molting, and post-molt phases, with effluent near 0.07 mg L⁻¹. Batch assays at ammonia saturation gave zero-order kinetics, apparent activation energies of 40.2 (95% CI 33.5–46.9) and 54.1 (46.6–61.6) kJ mol⁻¹, and temperature coefficients θ = 1.055 and 1.075. Tracer testing confirmed near-completely-mixed hydraulics, so the biofilm experienced the effluent concentration: the reactor oxygen-to-nitrogen ratio exceeded 60 g O₂ (g N)⁻¹, the ammonia-limited regime, in which tertiary MBBRs show first-order, transport-controlled behaviour and a near-unity apparent temperature coefficient. The batch coefficients therefore describe the biofilm at saturation, not the operational rate-limiting step. Dual controls attributed essentially all nitrification to the carrier biofilm. Saline MBBRs and crustacean recirculating aquaculture systems have each been studied separately; their combination supplies design parameters for a regime absent from freshwater and synthetic-feed datasets. Full article

Mycobacterium avium subspecies paratuberculosis (MAP) causes paratuberculosis (PTB), a chronic wasting enteritis of domestic and wild ruminants. Although the disease and/or its aetiology have been reported in livestock in the Sudan, they have not been investigated in wild animals. This study investigated MAP DNA prevalence in the Dinder National Park (DNP) to provide baseline data on PTB in wildlife. Fresh faecal samples (n = 274) of wild ruminants were collected from the ground around the six major water ponds of the DNP (44–48 samples/pond). Faecal DNA was extracted using a rapid extraction protocol and tested for MAP DNA using a real-time recombinase-aided amplification (RAA) assay. The vegetation structure around the ponds was observed and described. MAP DNA detection rates around the ponds ranged between 64.4% and 22.9%, with an overall apparent prevalence of 37.9% and a sensitivity/specificity-adjusted sample positivity of 42.4%. The faecal samples belonged to Reedbuck (Redunca redunca), Warthog (Phacochoerus aethiopicus), Waterbuck (Kobus defasa), African Buffalo (Syncerus caffer), Oribi (Ourebia ourebia), and among which MAP DNA detection rates were 39.4% (28/71), 39.1% (18/46), 38.0% (27/71), 37.5% (12/32) and 35.2% (19/54), respectively. The vegetation pattern around the ponds showed a notable difference that was consistent with MAP DNA detection. These results are consistent with the potential role of wildlife in paratuberculosis maintenance and transmission. Future investigations, including genotyping, would identify possible cross-transmission of the pathogen at the wildlife-domestic animal interface. Full article

Knowledge of farmed animals’ behavior and productive traits is essential to ensure the economic, environmental, and social sustainability of pastoral activities in a climate change scenario. However, limited information is available on breed differences under semi-extensive farming conditions. This study aimed to assess differences between Comisana and Sopravissana sheep in animal movement, body condition, and milk production. From June to October 2023, 40 female sheep were equipped with GPS-dataloggers to monitor movement patterns during free grazing. During the same season, Body Condition Score (BCS) was repeatedly assessed, while milk production was monitored at the breed-group level in 60 lactating ewes. The two groups showed similar movement patterns during free-grazing times. Milk production differed between the two groups and changed over time, with different temporal trajectories between groups; the Sopravissana group showed lower milk production than the Comisana group but a smaller overall reduction over time. Differences in body condition also emerged, with Sopravissana sheep maintaining a significantly higher BCS than Comisana sheep (p < 0.001). Post hoc tests confirmed significant breed differences in both physiological states (lactation/dry) and showed that lactating Sopravissana ewes maintained a higher BCS than dry Comisana ewes (p = 0.0002). These findings suggest that the differences observed in body condition and milk-production dynamics may be relevant when considering breed choice in Mediterranean pastoral systems and could contribute to improving sustainable farm management under increasing environmental challenges. Full article

Background: Caprine tuberculosis (cTB), caused by members of the Mycobacterium tuberculosis complex (MTBC) represents a significant concern for animal and public health. Antibody-based diagnostic tests have emerged as promising tools to complement ante-mortem official diagnostic techniques in ruminants, which mainly use serum, although alternative, easier-to-collect samples, such as milk, oral fluid, or feces, could also be used. The objective of this exploratory study was to evaluate the reactivity of an in-house P22 enzyme-linked immunosorbent assay (ELISA) in fecal samples from goats belonging to cTB-infected herds, using serum P22 ELISA and official diagnostic tests as comparators. Methods: Serum and fecal samples were collected from 51 and 42 adult female goats from a high-prevalence herd (n = 149) and a low-prevalence herd (n = 220), respectively, which defecated during sampling. These animals were subjected to official diagnostic techniques: single and comparative intradermal tuberculin tests and the interferon-gamma release assay. As no TB-free herd was available, a preliminary fecal cut-off was established by receiver operating characteristic analysis using serum P22 ELISA as the reference classification. Results: The percentage of positive animals in fecal samples from the high-prevalence herd was 68.6%, while no positive results were observed in the low-prevalence herd. Serum samples showed a higher number of positives in both herds compared to fecal samples (p < 0.05). The agreement between official diagnostic techniques and P22 ELISA (using serum or feces) was slight in general terms. When a combined classification based on all official tests was used as the reference, fecal P22 ELISA detected 58.0% of test-positive goats, none of them in the low-prevalence herd. Conclusions: Under the conditions of this field study (no TB-free controls or post-mortem confirmation of infection were available), fecal P22 ELISA showed limited reactivity compared with serum, particularly in the low-prevalence herd, suggesting limited usefulness for antibody detection in TB infected goats. Full article