Electrochemotherapy (ECT) is a local ablative treatment and in veterinary oncology, evidence supporting ECT efficacy is mainly derived from case reports and small case series, underscoring the need for standardized, multicentric data collection comparable to human oncology registries. The primary aim of this study was to establish VetInspECT, a veterinary clinical registry adapted from the human InspECT platform, to enable standardized reporting and multicentre data collection for ECT. The secondary aim was to evaluate antitumor effectiveness using data from the single centre that initiated the registry. VetInspECT was designed to systematically record patient characteristics, tumor features, treatment parameters, and follow-up outcomes in animals treated with ECT. Retrospective data spanning from 2012 to 2024 from feline patients treated at a Slovenian center were entered into the database. Squamous cell carcinoma (SCC) was the most frequently treated tumor (66%), followed by mast cell tumors (MCT; 12%) and other carcinomas and sarcomas. Complete response rates were highest in MCT (95%), followed by non-SCC carcinomas (90%), SCC (74%), and sarcomas (33%). Tumor size, clinical stage, anatomical location, and mucosal involvement significantly affected treatment response and progression-free survival. In conclusion, VetInspECT represents an important step toward standardized ECT reporting in veterinary oncology and supports the use of shared registries to enable robust multicentric analyses and broader clinical integration of ECT.
Stress is an umbrella term indicating increased allostatic load with the potential to reach a pathological state of the organism. In this context, cortisol concentration and neutrophil granulocytes-to-lymphocyte ratio (NEU: LYM) are expected to increase. On the functional pathway, stress reduces the oxidative burst capacity of neutrophils and therefore, the leucocyte coping capacity (LCC) quantifying this oxygen radical generation of neutrophils. Therefore, we hypothesized that in ponies subjected to repetitive handling, LCC will decrease and will be negatively correlated with cortisol concentration and NEU: LYM ratio. Thus, LCC was measured over 90 min by a portable chemiluminometer in nine university owned ponies involved in a vaccination study that included handling and sampling over 42 days. The area under the curve for LCC (AUCLCC) and LCC: NEU (AUCLCC: NEU), cortisol concentration measured by a validated EIA and NEU: LYM ratio were compared before and at days 1, 3, 28, 31 and 42 after immunization by linear mixed models. Pearson correlation coefficients were calculated between AUCLCC, AUCLCC: NEU, cortisol concentration and NEU: LYM ratio. AUCLCC, AUCLCC: NEU and cortisol concentration showed a significant decrease. AUCLCC was moderately positively correlated with NEU: LYM and weakly positively correlated with cortisol concentration, but no correlations were found for AUCLCC: NEU. The progressive decrease of AUCLCC, AUCLCC: NEU, and cortisol concentration might indicate an increasing stress load. However, this was not supported by the time course of NEU: LYM ratio or by the correlations between these parameters. While the slight decrease in LCC over time partially supported our hypothesis, we found no evidence of correlations among AUCLCC: NEU and the other measured stress parameters. This may indicate that different stress markers reflect distinct endpoints within the physiological pathways of the stress response. Future studies should confirm these preliminary results and also incorporate ethograms to obtain a more comprehensive understanding of the stress response and to further evaluate the value of LCC as a stress marker in horses.
Dermacentor reticulatus is a key tick species across Europe and an established vector of multiple pathogens affecting both human and animal health. Despite its significant role in disease transmission and its expanding distribution, genomic data for this species remain limited. Here, we present the first chromosome-scale genome assembly of D. reticulatus, constructed using Oxford Nanopore long-read sequencing. This chromosome-scale genome assembly revealed a repeat-rich genome, with approximately 63.6% of the total sequence consisting of repetitive elements. BUSCO analysis demonstrated strong genome completeness, with guided assembly achieving a score of 97.9%, comparable to related Dermacentor species. Gene annotation predicted 21,592 genes with a BUSCO completeness score of 98%. Functional characterisation, including Pfam domain assignment and gene ontology analysis, highlighted enrichment of molecular functions associated with detoxification, protease regulation, antimicrobial defence, and cholesterol metabolism. Additionally, the mitochondrial genome (15,103 bp), comprising 38 genes, was assembled, providing further insight into D. reticulatus phylogenetic placement in the Dermacentor genus. This genomic resource establishes a foundation for studying tick biology, evolution, and host-pathogen interactions.
Antimicrobials are currently an indispensable pillar of the animal production industry, relied upon to treat diseases that could otherwise severely impact animal welfare and productivity. However, overuse of antimicrobials is driving the selection of drug-resistant microbes. Coupled with the rising demands placed on animal production systems by the growing human population, we now face an increasingly urgent need for new strategies to address animal diseases and their impacts on productivity, efficiency, and welfare. The identification and development of alternatives to antimicrobials is therefore a critical and fast-growing field of research. The STAR IDAZ International Research Consortium recently conducted workshops that recruited international experts to develop research roadmaps aiming to stratify, organise, and clarify the ongoing research and outstanding questions that currently define the field of alternatives to antimicrobials. This review will explore the roadmap that deals with immunomodulators, a broad class of technologies that aim to beneficially alter host immunity in ways that promote animal health and growth, mitigate disease, and improve welfare at the human-animal interface. We will discuss the STAR IDAZ immunomodulators roadmap in detail, addressing the foundational biology underlying immunomodulation; the fields of research that link livestock immunity and metabolism with response to infections; and the immunomodulator technologies that are currently under development.
Residual feed intake (RFI) is an important indicator for measuring feed efficiency and can be used to screen sheep populations with low feed consumption and excellent growth performance. Its mechanism is closely related to the hypothalamus, which regulates animal feeding behavior, as well as the rumen and duodenum, which are responsible for the main digestive and absorptive functions. In theory, the hypothalamus-gastrointestinal axis is one of the key pathways affecting feed efficiency in ruminants. However, existing research has focused on the gastrointestinal microbiota and lacks systematic research on the interaction mechanism and functional expression differences between the hypothalamus and the gastrointestinal axis. The results indicate that Residual Feed Intake (RFI) significantly influences livestock quality in sheep. Regarding meat characteristics, the low-RFI (L-RFI) group exhibited a significantly higher cooking percentage in the longissimus dorsi muscle compared to the high-RFI (H-RFI) group. In terms of wool quality, L-RFI sheep were characterized by finer fiber diameters, shorter staple lengths, and an increased straight fiber rate. Mechanistically, RFI affected glutamatergic synapses, calcium signaling, and neurotransmitter receptor activity in the hypothalamus. Transcriptomic profiling further revealed that high-intake sheep showed upregulated lysosomal function, ATP metabolism, and cell cycle pathways in the rumen. Similarly, the duodenum of H-RFI sheep displayed marked activation of DNA replication and B-cell receptor signaling, alongside enhanced cholesterol transport and epithelial cell proliferation. In summary, this study, through a multi-dimensional analysis from the hypothalamus to the gastrointestinal tract, systematically reveals the physiological and molecular basis of RFI's influence on sheep feed efficiency, providing a new research paradigm for further exploring the role of the brain-gut axis in the regulation of animal feed efficiency.
In Central Uganda, smallholder farmers rely on indigenous ethnoveterinary botanicals as phytogenic feed additives (PFAs) for indigenous chickens (Gallus gallus domesticus). However, the balance between their nutritional benefits and toxicological risks remains poorly characterized. Following an ethnoveterinary survey in Najjembe sub-county where Capsicum frutescens (ranked 2nd ), Cannabis sativa (3rd ), and Nicotiana tabacum (4th) were prioritized by farmers, this study investigated their nutritional and safety profiles. Aloe vera (1st ) was excluded from analysis due to extensive prior documentation. Samples were analyzed for proximate, mineral, and antioxidant profiles using AOAC Official Methods. Antinutrients (tannins, oxalates, phytates) and heavy metals (Pb, Cd) were quantified using vanillin-HCl spectrophotometry and Atomic Absorption Spectrophotometry, respectively. Results showed significant nutritional variation among species (p < 0.05). C. sativa contained the highest crude protein (22.20 ± 1.1%) and was exceptionally rich in calcium and potassium. C. frutescens exhibited the highest caloric density (452.9 kcal/100 g) and antioxidant activity (IC50 = 11.78 mg/mL). Conversely, N. tabacum had the highest iron and magnesium but also the highest antinutrient concentrations. Critically, lead (Pb) and cadmium (Cd) levels in N. tabacum and C. sativa significantly exceeded FAO/WHO safety limits (Pb > 0.3 mg/kg), resulting in hazard indices (HI > 1.0), signaling potential health risks for chickens and consumers. While these plants offer high nutraceutical value, the heavy metal bioaccumulation in N. tabacum and C. sativa necessitates strict inclusion limits and processing to ensure food safety. C. frutescens is recommended as the most viable PFA for feed integration.
An emerging disease known as mule deerpox virus (Cervidpoxvirus muledeerpox) (DPV) has spread across North America in both wild and captive cervid populations. While little is known about its prevalence in wild cervids, research in domestic populations indicates a high mortality associated with this pathogen, particularly in fawns. Most recently, a new Cervidpoxvirus has been reported in semi-domestic populations of reindeer in Sweden and Norway, expanding its geographical range internationally. Currently, there is no standard protocol to detect this virus, and the increasing range of this pathogen highlights the importance of a standard validated diagnostic test. In this study, we developed and partially-validated a TaqMan quantitative PCR assay to detect Cervidpoxvirus based on the major capsid protein using the World Organization for Animal Health's diagnostic assay guidelines. Aligning the three available whole genomes of DPV from GenBank W-848-83 (GenBank: NC_006966.1); W-1170-84 (GenBank: AY689437.1); MDPV-F (GenBank: MF966153), showed that the major capsid protein gene was highly conserved (> 99%) and primers and probe were designed to amplify a 83 bp region of this gene with 100% agreement. Analytical performance estimated that the assay was exclusive for DPV and highly sensitive (limit of detection ≥ 10 DNA copies), while diagnostic performance found that the assay was 100 times more sensitive than a standard conventional PCR assay. With this assay, detection of Cervidpoxvirus will be faster and more efficient than current methods, allowing both researchers and veterinarians to diagnose and prevent the further spread of this disease.
Monkeypox (Mpox) is a zoonotic disease that threatens global public health. Different clades of monkeypox virus (MPXV) vary in transmissibility and pathogenicity. In 2023, a Clade Ib MPXV variant emerged in the Democratic Republic of the Congo, continued to spread in parts of Africa, and subsequently spilled over to other regions, posing new challenges for outbreak prevention and control. We established stable mouse models of MPXV Clade Ib infection by intranasally infecting C57BL/6/STAT1-/-, AGB6 (C57BL/6-Ifngr1-/-Ifnar1-/-), C57BL/6, and BALB/c mice. Susceptible strains showed marked body weight loss, high viral loads in tissues, and severe histopathological lesions. RNA-seq analysis of spleens at the early stage of infection showed that differentially expressed genes were mainly enriched in interferon-mediated antiviral pathways and inflammatory cytokine-related pathways, whereas genes associated with adaptive immune responses were downregulated. Comparative analysis showed that MPXV Clade Ib caused more severe disease phenotypes than Clade IIb under the same experimental conditions, which is consistent with reported differences in clinical severity. We established reproducible mouse models for MPXV Clade Ib infection and demonstrated that Clade Ib showed greater replication capacity and pathogenicity than Clade IIb in these models. This study also provides a foundation for subsequent research on the pathogenesis of MPXV and the evaluation of antiviral efficacy.
Rift Valley fever virus (RVFV) is an emerging zoonotic virus of major public health and veterinary concern across Africa. Although past genomic studies have focused on outbreak response and lineage classification, the molecular mechanisms driving viral persistence and adaptation remain poorly understood. This study aimed to identify RVFV protein-coding mutations and adaptive signatures across multiple epidemics and epizootics in Africa. This retrospective genomic analysis examined 596 RVFV segment sequences retrieved from the NCBI Virus database, including L (n = 173), M (n = 196) and S (n = 227) sequences from 13 African countries across human and non-human hosts between 1944 and 2022. Following the identification of protein-coding mutations via Genome Detective and custom scripts, we performed phylogenetic reconstruction using IQ-TREE and host-state reconstruction to map cross-species transmission patterns and selection pressure analyses were conducted using codon-based models implemented in the Datamonkey platform. All data analyses and visualizations were performed using R software. A total of 7,339 protein-coding mutations were identified, ranging from 2-20 per isolate. RVFV isolates collected in South Africa, Kenya and Madagascar exhibited the highest genomic diversity. Comparative analysis revealed higher mutation burdens in the L and S segments than in the M segment, with broader diversity among non-human hosts. Phylogenetic reconstruction showed that human-derived sequences clustered within livestock and vector lineages, a pattern consistent with significant genetic bottlenecks during spillovers. Notably, host-state reconstruction identified livestock lineages as the primary source of human outbreaks. We identified seven recurrent amino acid mutations across the genome: N277S, N277D and S278N in the polymerase (L); I442S, I442V, V659A in the glycoproteins (M); and N133S in the NSs protein (S). FUBAR-supported signals consistent with diversifying selection were identified at corresponding codon sites, particularly within the polymerase and glycoprotein regions, highlighting candidate residues potentially associated with adaptive processes affecting replication efficiency and immune evasion. Our findings demonstrate that RVFV evolution across Africa is geographically and temporally heterogeneous, with livestock infections identified as the primary driver of human outbreaks. While RVFV evolution is largely shaped by purifying selection, FUBAR analysis revealed a limited number of candidate codon sites under diversifying selection that may facilitate host-specific adaptation. These host-specific pressures likely contribute to adaptive substitutions that fine-tune polymerase function, alter glycoprotein antigenicity and enhance immune escape. Collectively, these results reveal the molecular mechanisms underpinning viral persistence and provide information to support the design of cross-protective vaccines.
Staphylococcus hyicus is recognized as one of causative agents of porcine exudative epidermitis in piglets. However, research on clinical pathogenic S. hyicus remains limited. In this study, multidrug-resistant S. hyicus was isolated from a large-scale pig farm with cases of fatal exudative epidermitis in piglets. By characterizing the phenotypes and genotypes of S. hyicus isolates, we provide insights for clinical management of exudative epidermitis. Two 21-day-old piglets with clinical signs underwent necropsy and histopathological examination. 48 samples from lactating sows and suckling piglets were collected for pathogen identification. Antimicrobial and disinfectant susceptibility of the isolates was determined using broth microdilution, as well as whole-genome sequencing (WGS) was used to identify antimicrobial resistance genes. Moreover, comparative genomic analysis with public genomes in the Genbank database was performed. The virulence of mecA-carrying S. hyicus was evaluated using the Galleria mellonella infection model. Exudative epidermitis was first observed in 3-day-old piglets. Through necropsy, pathogen isolation, and other diagnostic approaches, S. hyicus was preliminarily identified as the primary causative agent of this episode, and 23 S. hyicus were isolated from samples. Antimicrobial susceptibility testing indicated most of isolated S. hyicus were susceptible to doxycycline, vancomycin and linezolid, while resistant to florfenicol, erythromycin, spectinomycin, amoxicillin, ceftiofur, enrofloxacin that are commonly used for pigs. Twelve resistance genes were identified by WGS, including aadD, ant(6)-Ia, aph(2'')-Ia, blaZ, erm(B), erm(C), fexA, lnu(B), lsa(E), mecA, tet(L), and tet(M). Compared with 39 GenBank genomes, mecA-carrying S. hyicus in our study carried more resistance genes and exhC islands closely related to previous Chinese strains. Based on our findings, we implemented a targeted treatment protocol that brought the outbreak under control and reduced piglet mortality to below 5%. This study successfully isolated pathogenic mecA-carrying S. hyicus responsible for high mortality in piglets, characterized its drug resistance phenotypes and genotypes, offering valuable insights for the prevention and control of this pathogen in piglets.
Ticks are major ectoparasites that significantly impact camels in Ethiopia. The local climate facilitates widespread infestation, particularly among camels, which are vital resources in East Africa. This study investigated pooled infestations of ixodid ticks in Ethiopian camels. In accordance with the PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses)-2020 guidelines, a comprehensive search was performed across multiple electronic databases, including Scopus, ScienceDirect, African Journal Online, PubMed, Web of Science and the Wiley Online Library. The Joanna Briggs Institute (JBI) checklist for prevalence data was used to assess the quality of included studies. Statistical examination was executed via Stata v.14. To calculate the overall infestation rate, a random-effects model was applied and established a 95% confidence interval for the data. To assess the consistency of the data, heterogeneity was measured using the I2 statistic, with further exploration conducted via Galbraith plots and subgroup and sensitivity analyses. To evaluate the presence of potential publication bias, the study utilized visual inspections of funnel plots complemented by the statistical rigor of Egger's test. Ultimately, 11 cross-sectional studies met the inclusion criteria, totaling 4,895 animals (camels) across six regions of Ethiopia. The meta-analysis in this review revealed an overall pooled infestation rate of ixodid tick infestations of 85.68% (95% CI: 78.29-93.06). Regional analysis revealed that the greatest percentage of infestations was in Tigray (96.6%), Dire Dawa (94.0%) and Afar (90.2%), whereas the lowest percentage of infestations was recorded in the Amhara region (55.2%). The taxonomic identification revealed the presence of three tick genera, specifically Rhipicephalus (Boophilus), Amblyomma, and Hyalomma. Rhipicephalus (Boophilus) was the most dominant genus, with an infestation rate of 56.44%, whereas Amblyomma was the least common (14.65%). The current study revealed that infestations decreased from 90.18% from 2013 to 2019 to 82.03% from 2020 to 2025. In Ethiopia, ixodid ticks represent a major category of ectoparasites that persistently compromise the health and economic productivity of camel populations. Tick control actions must target the most common genera and the most affected geographic regions. Future research should shift toward investigating breed-specific susceptibility and the development of sustainable, integrated control strategies.
Canine lymphoma is a common malignancy with an annual incidence rate of 160 cases per 100,000 dogs. The primary therapeutic protocol in clinical use is treatment with cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP); however, relapses remain frequent, and early detection of recurrence is challenging. IDEXX Cancer Dx™ testing is a commercially available, blood-based test developed for the detection of canine lymphoma and may also have utility as a minimally invasive tool for longitudinal monitoring during treatment. Ten client-owned dogs with cytologic confirmation of lymphoma, immunophenotyping by flow cytometry and/or genotypic clonality testing by PCR for antigen receptor rearrangement (PARR), and an initial response to CHOP therapy were enrolled in this prospective longitudinal study. Whole blood was collected serially during chemotherapy and analyzed using Cancer Dx testing, with results reported as positive or negative. Clinical response was evaluated using the Veterinary Cooperative Oncology Group (VCOG) response evaluation criteria for peripheral nodal lymphoma in dogs. Cancer Dx test results showed concordance with clinical disease status (Somers' D = 0.70, 95% CI [0.59, 0.82]; Kendall's τ-b = 0.67, 95% CI [0.56, 0.78]). Among the ten cases, test results for eight (80%) dogs became negative, corresponding with clinical remission. Seven of ten (70%) dogs maintained negative test results during complete remission; five of ten (50%) dogs became positive at clinical relapse. Discordance between Cancer Dx testing results and clinical status at the time of initial clinical remission occurred in 2 out of 10 (20%) of the enrolled dogs. Two dogs (Dogs D and I) remained positive despite achieving clinical remission during the initial weeks of CHOP therapy, suggesting delayed or incomplete biomarker conversion. Serial Cancer Dx testing results were concordant with VCOG Response Evaluation Criteria-defined clinical status. While these results support further investigation of Cancer Dx testing as a monitoring tool during chemotherapy for lymphoma, the current performance of the assay is insufficient for reliable, stand-alone monitoring without further refinement.
Understanding the genetic architecture of economically important traits in poultry is critical for improving breeding strategies. In this study, we investigated a backcrossing scheme between a White Layer line and Araucana chickens. Genome-wide association studies (GWAS) were performed using array-genotyped and imputed data. We also explored the use of correlated traits as covariates, which helps to distinguish between pleiotropic and trait-specific associations. We applied two Bayesian fine-mapping methods to refine GWAS-identified QTLs and pinpoint candidate variants and genes associated with egg number (EN, from 20 to 71 weeks), egg weight (EW, from 30 to 70 weeks), and body weight (BW, at 32 weeks): the forward selection approach and functional annotation enrichment implemented in BFMAP, and the shotgun stochastic search algorithm implemented in FINEMAP. GWAS identified multiple significant loci associated with BW and EW, with high heritability estimated for these traits. EN showed a more polygenic architecture with lower heritability across most periods. Including correlated traits as covariates in GWAS revealed pleiotropic loci, particularly on chromosomes 1 and 4, that influenced both BW and EW, as well as loci specific to individual traits. Both fine-mapping methods successfully pinpointed candidate genes such as NCAPG, LCORL, and IGF2BP1, which are well known for their roles in growth and body size across species. Several novel candidate genes were also highlighted for EN. Notably, some fine-mapped results reflected patterns consistent with the covariate-adjusted GWAS results. This study demonstrates the power of combining GWAS with imputation and fine-mapping methods in chickens to uncover the genetic basis of economically important traits. Furthermore, incorporating correlated traits as covariates in GWAS provided valuable insights, enabling the distinction between pleiotropic and trait-specific loci. Together, these approaches refine GWAS signals and deepened our understanding of the genetic architecture underlying complex traits.
Myofascial pain syndrome (MPS) is considered highly prevalent in horses, yet it is frequently underdiagnosed due to limited understanding of its clinical presentation and functional consequences, as well as the inherent challenges associated with its clinical assessment and objective measurement. In equines, the relationship between palpation-based MPS indicators, gait asymmetries, and pain-related behaviours during ridden work remains unclear. The primary aim of this study was to evaluate the association between thoracolumbar palpation pain scores and gait asymmetries measured in-hand and during ridden exercise, as well as behavioural indicators of pain assessed using the ridden horse pain ethogram (RHpE). The secondary aim was to explore the effect of a rider on gait asymmetry using a within-horse comparison between in-hand and ridden conditions. Clinical and behavioural signs and demographic variables were also described. Twenty police working horses from the Special Mounted Unit of the Spanish National Police in Madrid were included. At least one myofascial trigger point, identified by a palpable taut band and a hypersensitive spot, was identified in all horses (100%) in the thoracolumbar region, followed by the jump sign (85%) and restricted range of motion (80%). Local twitch responses were not observed in any horse (0%). Clinical indicators of MPS were not associated with objective gait asymmetries measures in either condition. Forelimb push-off asymmetry (HDmax) increased under ridden conditions (β = 0.196, 95% CI 0.028-0.034, p = 0.022). Training level modified this effect, with higher-level horses exhibiting greater increases in HDmax when ridden. The mean RHpE score was 7.2, with 40% of horses scoring ≥ 8 behaviours. The most frequent behaviours were ears rotated back (95%), mouth opening (95%), and intense stare (80%). No significant associations were found between RHpE scores and either MPS clinical signs or ridden gait asymmetries. This is the first study integrating palpation-based assessment of thoracolumbar MPS with objective gait analysis and the RHpE. The findings suggest the importance of palpation as the primary diagnostic approach for MPS in horses and indicate that a multimodal approach may improve the understanding and management of equine MPS.
Natural products with dual immunomodulatory and antimicrobial functions offer promising strategies to reduce antibiotic use in livestock. Glycyrrhizic acid (GA), the principal bioactive component of licorice, has demonstrated anti-inflammatory and antiviral properties, yet its translational potential in swine health remains underexplored. This study evaluated the efficacy of GA in weaned piglets under commercial nursery conditions as an antibiotic alternative. A total of 225 weaned piglets were assigned to five groups: negative control (CON, basal diet), farm routine (FA, conventional antibiotics), and three GA-supplemented groups (GLL, 0.65 g/kg; GLM, 1.3 g/kg; GLH, 2.6 g/kg). The result showed that dietary GA supplementation (2.6 g/kg) numerically improved growth performance and reduced cough scores, although not statistically significant. GA significantly decreased the diarrhea index and improved skin scores. GA also significantly increased serum IgG and IgM levels in piglets and showed a trend toward higher IgA levels. Furthermore, GA exhibited a trend toward lowering serum IL‑1β levels while upregulating IFN‑γ and IL‑10 levels. Regarding antioxidant parameters, GA significantly upregulated T‑SOD, GSH‑PX, and CAT activities and downregulated LDH activity. Metagenomic analysis revealed that high‑dose glycyrrhizic acid (GA) significantly increased the abundance of Alloprevotella, while decreasing the abundances of Moraxella pluranimalium and 11 other pathogenic species associated with respiratory diseases and lung injury, including Glaesserella parasuis, Mesomycoplasma hyorhinis, Mesomycoplasma hyopneumoniae, Streptococcus suis, among others, thereby reshaping the upper respiratory tract microbiota of pigs. Collectively, these findings support GA as a viable non-antibiotic strategy for improving immune function, antioxidant capacity, and respiratory health in weaned piglets.
Salmonella spp. is one of the main causes of food-borne zoonotic infections in the European Union with Enteritidis being the most reported serovar. Salmonella is commonly associated with domestic animals, although it has also been associated with wildlife. The domestic goat (Capra hircus) appears to have a relatively low susceptibility to this pathogen. Salmonella has been described in game meat of different wild ruminant species; however, limited information is available regarding the susceptibility of wild goats, which may represent a source of human infection.In this study, we describe the epidemiological, molecular, microbiological and pathology findings of an unusual outbreak of clinical salmonellosis in captive Iberian wild goat (Capra pyrenaica) in Spain. The outbreak, plausibly caused by S. Enteritidis (9,12:g, m:-), was characterized by abortions, neonatal mortality and sporadic cases of septicaemic salmonellosis in adult wild goats. Most isolates showed the same Minimum Inhibitory Concentration profiles using the Thermo Scientific™ Sensititre™ EU Surveillance Salmonella/E. coli EUVSEC3 AST panel. Seven isolates were sequenced and classified as ST11 showing the same set of 140 virulence genes. The genomes differed by 0-1 SNP (NC_011294.1:g.2862805G > A).Due to the unusual circumstances of the outbreak, the origin of the outbreak was also investigated through non-invasive sampling for nucleic acid detection using sponges pre-hydrated with an isotonic surfactant liquid and for bacteriology. Salmonella was exclusively detected from wild-goat enclosures, allowing to presumably rule out other potential animal-sources of infection, including peridomestic/wild animals such as pigeons, rats, rabbits and partridges present and analysed in the farm surroundings, although another untested animal source could still be involved.The unusual presentation of this Salmonella outbreak could be explained by the convergence of epidemiological and ecological factors (infection origin linked to peridomestic animals or humans, management conditions which may favour pathogen emergence, potentially increased susceptibility of wild ruminants to infection or other factors which may be involved). The non-invasive sampling approach facilitated Salmonella surveillance in this underexplored wildlife host.
In the previous single-cell transcriptome analysis of goat ovulation, we identified that CACNA2D1 may play a critical role in immune cell infiltration during the ovulation process. Therefore, this study aimed to investigate the function of CACNA2D1 in ovulation. CACNA2D1-associated signaling was pharmacologically inhibited in a mouse ovulation model to evaluate ovarian immune cell infiltration, ovulatory output, and transcriptomic changes. In vitro, CACNA2D1 knockdown and overexpression were performed in ovarian granulosa cells to assess chemokine production, intracellular calcium signaling, CaMKII activation, and cell proliferation. Granulosa cell-mediated macrophage recruitment was functionally examined using a RAW264.7 transwell migration assay. CACNA2D1 showed increased expression during the periovulatory period, and pharmacological inhibition of CACNA2D1-associated signaling in mice significantly reduced ovarian immune cell infiltration, particularly macrophages, accompanied by decreased ovulatory output. Ovarian transcriptomic analysis after inhibition revealed broad alterations in genes related to immune regulation, signal transduction, and cellular functional states. In granulosa cells, CACNA2D1 knockdown significantly reduced intracellular Ca2+ levels, decreased CaMKII phosphorylation, and suppressed the expression and secretion of immune recruitment-related factors, including IL6, CCL2, and CXCL12. Restoration of calcium signaling with A-23,187 rescued the reduced expression of these chemokines. Functionally, CACNA2D1 knockdown impaired granulosa cell-mediated RAW264.7 macrophage migration in a transwell co-culture system. CACNA2D1 also influenced granulosa cell proliferative activity, whereas its effect on macrophage-related responses appeared to be more closely associated with chemotactic signaling than with macrophage proliferation. This study identifies CACNA2D1 as a periovulatory granulosa cell-associated regulator involved in calcium-dependent chemokine production and macrophage recruitment. These findings provide new insight into ovarian immune remodeling during ovulation and offer a potential molecular basis for future studies of follicular development, ovulation efficiency, and fertility regulation in livestock species.
Duck hepatitis virus (DHV) continues to pose a substantial threat to duck production worldwide, causing acute hepatitis, neurological manifestations, and elevated mortality rates, even in vaccinated flocks. During 2022-2023, a significant outbreak involving DHV-1 and DHV-3 was reported in duck farms across five governorates in North Egypt. The outbreaks primarily affected Pekin ducklings aged 4-15 days, with mortality rates ranging from 50% to 70%. Affected ducklings exhibited characteristic pathological lesions, including hepatomegaly with haemorrhages, splenomegaly, and renal enlargement. A total of 30 liver and spleen samples collected from affected farms were analysed using reverse transcription PCR (RT-PCR) targeting the 3' untranslated region (3'UTR) and the VP1 gene. DHV was detected in 56.7% (17/30) of the samples. Among the positive cases (n = 17), DHV-1 was identified in 29.4% (5/17), whereas DHV-3 accounted for 70.6% (12/17). Phylogenetic analysis based on whole-genome sequencing revealed that DHV-1 strains clustered within sub-clade 1a, demonstrating high genetic similarity (99.2-99.8%) with previously reported Egyptian isolates and Chinese reference strains (e.g., DHAV-1-CH-2012). In contrast, DHV-3 strains grouped within sub-clade 3a and showed close genetic relatedness to Chinese strains (97.1-98.7%), while showing marked divergence (75.9-76.3% nucleotide identity) from currently used vaccine strains. Genetic analysis identified multiple mutations in both viral types. DHV-1 exhibited amino acid substitutions within VP0, VP3, and non-structural protein regions, alongside five mutations in hypervariable region 1 (HVR1) and one mutation in HVR2 of the VP1. DHV-3 demonstrated distinct mutations within the VP1, including a unique E681K substitution identified in the Egyptian strain ND3. The findings demonstrate ongoing genetic evolution of DHV circulating in Egyptian duck farms, characterized by the predominance of genetically distinct DHAV-3 strains, with significant divergence from currently used vaccine strains. This genetic divergence may contribute to altered vaccine performance and continued disease outbreaks; however, its impact on vaccine-induced protection requires further experimental validation. Continuous genomic surveillance and the development of updated, strain-matched or multivalent vaccines are therefore essential to control DHV infections and minimise economic losses in Egypt's duck industry.
Artificial insemination (AI) success in Murciano-Granadina goats is influenced by a complex interaction of male, female, management, and environmental factors. This study aimed to identify the main drivers and threshold conditions affecting fertility outcomes in commercial AI programs using long-term field data. A dataset of 3,122 inseminations performed between 2010 and 2019 was analyzed using canonical discriminant analysis and CHAID decision trees. Multicollinearity screening excluded 24 redundant predictors out of an initial set of 52 variables, retaining 28 variables with the greatest explanatory power. The canonical model showed that the first six functions explained 72.4% of total variance, with barometric pressure, temperature, and rainfall at insemination as dominant environmental loadings. Fertility increased when minimum temperatures exceeded 14.7 °C but declined below 9 °C, coinciding with reduced semen quality. Wind speeds above 2.9 m/s and humid easterly winds reduced conception probability by up to 18% due to potential stress-inducing effects at semen collection. Altitudes between 451 and 720 m were associated with the highest fertility rates. Management thresholds were also important: fertility declined when inseminated group size exceeded 63 does, whereas herds larger than 50 animals showed improved synchrony and higher conception probabilities. Female age strongly influenced outcomes, with immature does (~ 2 years) more frequently associated with low fertility, while multiparous females (> 3 parturitions) achieved fertility rates up to 15% higher. Bucks aged 3-5 years produced semen with superior fertility outcomes compared with younger (1-3 years) or older (> 7 years) males. Somatic cell counts below 3,000 × 103/mL increased the probability of high fertility outcomes, whereas counts above 4,000 × 103/mL were associated with markedly reduced success. Compensatory outcomes were also observed, where acceptable fertility occurred despite low semen quality under favorable physiological and management conditions. AI fertility in Murciano-Granadina goats results from multifactorial interactions among environmental conditions, physiological status, and herd management. Identifying key environmental, physiological, and management thresholds may contribute to improved planning of AI programs and may help guide future strategies aimed at enhancing reproductive efficiency.
Greece holds a critical geographic position in the introduction of domestic sheep into Europe during Neolithic and post-Neolithic migrations, acting as a bridge with Asia. The genomic profile of Greek sheep remains underexplored compared to global breeds, despite their importance for sustainable livestock production. In this study, we constructed an extensive 50 K SNP data collection from 1,625 Greek indigenous sheep reared in 23 flocks compared to southwest Asian, Asian, south, central and north European populations. We detected a diverse genomic profile for Greek breeds linked to both European and Asian ancestry using PCA, admixture and NeighborNet analyses. The Greek breeds were clearly differentiated at the breed level with close genetic relationships between Greek mainland populations and the Italian Zackel-type sheep. PCA clustering and network topology showed an east-to-west genetic axis extending from Asia to the Iberian Peninsula across the Mediterranean, consistent with the Mediterranean dispersal of domestic sheep, with Greece serving as an interlink between the two continents. The Greek cluster showed the highest reticulation level in the regional phylogenetic analysis, implying recent admixture events compared to other geographic clusters. The average observed heterozygosity in Greek sheep (Ho = 0.358), ranging from 0.326 to 0.376, was comparable to that of Mediterranean breeds (Ho = 0.366). At local scale, the analysis of multiple farms revealed heterogeneous management practices, enabling genetic identification in particular cases as evidenced by the admixture profiles by both between- and within-breed comparisons. Semi-fat-tailed Chios (FIS= 0.17) and Pelagonia (FIS= 0.10) sheep displayed higher inbreeding across farms compared to the other Greek sheep attributed to long-lasting selection pressure, conservation efforts and sampling scheme, respectively. Collectively, our findings can be integrated into national conservation actions, especially for endangered populations, to prevent genetic erosion and support sustainable management of local livestock genetic resources.