Veterinary Sciences doi: 10.3390/vetsci13070715

Authors: Kyosuke Takeuchi Kenji Hosoya Ryo Owaki Ryohei Kinoshita Sangho Kim Masahiro Okumura

Gastrinoma is a rare neuroendocrine tumor in dogs that arises from functional pancreatic non-β cells and secretes excessive gastrin. Gastrinoma in dogs has a high metastasis rate at diagnosis, and curative treatment is often difficult. There are few reports describing the long-term outcomes of surgical treatment in dogs with gastrinoma, and no established method for monitoring disease progression has been reported. We report here the cases of two adult dogs with pancreatic gastrinoma treated with surgical resection. Pharmacological therapy using proton pump inhibitors was administered to control gastrointestinal symptoms caused by hypergastrinemia. Multiple surgeries were performed to reduce the volume of primary and metastatic gastrinoma lesions. Serial serum gastrin concentrations were used as an adjunctive monitoring marker during follow-up. Changes in serum gastrin concentrations were used to support decisions regarding further imaging. Additional surgery was performed based on serum gastrin concentrations, clinical signs, and diagnostic imaging findings. A total of four surgeries were performed in Case 1 and three surgeries in Case 2. Both dogs maintained well-controlled gastrointestinal symptoms and showed long-term survival. The changes in serum gastrin concentrations in these cases suggest that serial serum gastrin measurement may be useful for monitoring recurrence or tumor progression in canine gastrinoma.

Veterinary Sciences doi: 10.3390/vetsci13070714

Authors: Iosif Vasiu Mădălina Florina Dragomir Ciprian Andrei Ober Cosmin Petru Peştean Romelia Pop Marian Taulescu Claudiu Gal Seppo Saari David Vicente

Four client-owned dogs with histopathologically confirmed periorbital tumors were included in this retrospective case series: one spayed female, one intact male, and two neutered males. The dogs ranged in age from 6 to 8 years. The most common clinical signs were slowly progressive exophthalmos (4/4 dogs), epiphora (3/4 dogs), and prolapse of the nictitating membrane gland (2/4 dogs). Periorbital disease was initially suspected based on the clinical examination. Computed tomography (CT) was subsequently used to characterize the presence, location, and extent of the lesion and for surgical planning, whereas histopathological examination confirmed the tumor type. The identified tumors included two multilobular tumors of bone (MTB), one inflammatory myofibroblastic tumor (IMT), and one sebaceous carcinoma (SC). Treatment for the first three dogs consisted of wide tumor excision via combined modified lateral orbitotomy and partial ostectomies of the zygomatic arch and mandibular ramus. The remaining dog received conservative therapy. For the first two surgically treated dogs, survival times were 55 months; the third dog was lost to follow-up two weeks after surgery. For the conservatively treated dog, the survival time was 4 months. Although limited by the small number of heterogeneous cases, this report suggests that the described surgical approach facilitated en bloc excision in these three surgically treated dogs. However, histologically tumor-free surgical margins were confirmed in only one of the three surgically treated dogs, and clinical outcomes varied according to tumor type, extent of disease, and duration of follow-up. Therefore, further studies are required before definitive conclusions regarding oncologic outcome or prognosis can be drawn.

Veterinary Sciences doi: 10.3390/vetsci13070713

Authors: Lauren Chambers Wiebke Jansen Dušan Palić Andrea Fabris Alain Schonbrodt Leona Nepejchalová Nancy De Briyne

Availability of veterinary medicinal products is a critical determinant of health, welfare, and economic sustainability in European aquaculture. This study assessed the changes in the authorization and reported market availability of veterinary medicines for finfish in the European Union (EU) and European Free Trade Association (EFTA) between 2018 and 2026, by comparing and analyzing data on veterinary medicinal products for aquatic species from the Heads of Medicines Agencies (HMA), the FishMedPlus Coalition, and the European Medicines Agency Union Product Database. The results revealed a wide divergence between regulatory authorization and reported availability. While the total number of authorized VMPs nominally increased from 304 in 2018 to 332 in 2025 (+9.2%), the number of reported unique active substances available declined (−16.7%). Crucially, the analysis of 2025 data indicates that only 26.5% of authorized products were reported to be actively available on the market, creating a “paper market” that masks severe therapeutic gaps, particularly in minor fish species such as seabass, seabream, carp, and trout sectors. Potential key drivers identified were limited markets, industry consolidation, increased burden related to stricter environmental requirements, as well as enlarged administrative and financial burdens. Despite EU legislative mechanisms designed to support limited markets, the sector faces stagnation in therapeutic diversity, necessitating continued reliance on the off-label use of medicines.

Veterinary Sciences doi: 10.3390/vetsci13070712

Authors: Natthrit Roekngam Sunisa Khongthong Ricadonna Raissa Bongkoch Chonglomkrod Sara Niae

Bacterial and fungal skin infections are common in companion animals and often require antimicrobial therapy, contributing to antimicrobial resistance. This study developed shampoos containing red palm oil (RPO) and extracts of Rhinacanthus nasutus (RN), Curcuma longa (CL), Zingiber montanum (ZM), and Zingiber officinale (ZO), which provide antibacterial, antifungal, antioxidant, and anti-inflammatory activities. Extracts were obtained using MAE and were evaluated for phytochemical composition, total phenolic content, antioxidant activity (DPPH assay), anti-inflammatory activity (COX-2 inhibition), and antimicrobial activity. RPO contained high levels of vitamin E and β-carotene. Rhinacanthin, curcumin, terpinen-4-ol, and 6-gingerol were the major phytochemical constituents of RN, CL, ZM, and ZO extracts, respectively. All extracts significantly reduced COX-2 expression (p < 0.05), while RN exhibited the strongest antioxidant activity (DPPH IC50 = 15.23 ± 0.53 µg/mL). RN and ZM demonstrated the strongest antibacterial activity against Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MIC = 8 µg/mL; MBC = 8–16 µg/mL), while ZO exhibited the strongest antifungal activity against Malassezia pachydermatis and Microsporum gypseum (MIC = 2, 2 µg/mL; MFC = 2, 16 µg/mL). The developed shampoos containing RPO and the four herb extracts showed suitable pH values, stable viscosity, antibacterial inhibition, and antifungal inhibition (p < 0.05).

Veterinary Sciences doi: 10.3390/vetsci13070711

Authors: Ji-Hyun Park Geon-Ung Byun Joon-Woo Lee Dae-Hyun Kim Dong-In Jung Dongbin Lee Chang-Hwan Moon

Acquired myasthenia gravis (MG) is an autoimmune disorder of the neuromuscular junction caused by antibodies against the acetylcholine receptor, leading to generalized muscle weakness and, in severe cases, respiratory muscle paralysis. While MG is well described in dogs, it is rarely reported in cats, and fulminant cases with rapid progression have a poor prognosis. An 11-year-old neutered female Persian cat presented with acute hindlimb weakness, which progressed to tetraplegia within 4 days. The clinical course initially resembled acute polyradiculoneuritis, and electromyography revealed positive sharp waves and fibrillation potentials. Diagnostic imaging identified a cystic cranial mediastinal mass without megaesophagus. Worsening respiratory muscle paralysis prompted surgical excision of the mass via left intercostal thoracotomy. Postoperatively, the cat developed persistent respiratory failure requiring re-intubation, mechanical ventilation, and temporary tracheostomy. Histopathological and immunohistochemical examination confirmed a thymoma, and serum acetylcholine receptor antibody levels were markedly elevated, establishing the diagnosis of thymoma-associated fulminant MG. Despite tumor excision and medical therapy with pyridostigmine and prednisolone, the cat remained ventilator-dependent and was euthanized on postoperative day 3. This case emphasizes early recognition of respiratory compromise, prompt diagnostic confirmation, intensive ventilatory support, and consideration of adjunctive immunomodulatory therapies.

Veterinary Sciences doi: 10.3390/vetsci13070710

Authors: Ding Li Xiaoping Wu Fengfang Zhou Jiacheng Zhang Kuncan Wei Yulin Lu Fangyu Yuan Weiqing Huang

Microbial communities associated with fish mucosal tissues play important roles in host health and environmental adaptation. However, the effects of contrasting aquaculture conditions on the microorganisms of the large yellow croaker (Larimichthys crocea, L. crocea) remain poorly understood. In this study, 16S rRNA gene sequencing was used to compare the gut and gill microbiota of L. crocea cultured in a marine system and a saline–alkaline system, together with the corresponding surrounding water samples. The results showed that both host tissue type and rearing environment significantly influenced microbial community structure. Water samples generally exhibited higher alpha diversity than host-associated samples, whereas the gut and gill communities were clearly separated from the surrounding water microbial community. The intestine showed stronger culture system differentiation than the gill, suggesting greater niche selectivity, while the gill retained a closer association with the rearing water. Across multiple taxonomic levels, microbial composition differed markedly among water, gut, and gill samples, and the divergence between the marine culture and saline–alkaline culture system became more pronounced at finer taxonomic resolution. Differentially enriched taxa further revealed clear tissue- and habitat-associated microbial patterns. In addition, several dominant taxa were significantly correlated with environmental variables, especially salinity and nutrient-related factors, indicating that physicochemical heterogeneity contributed substantially to microbial community assembly. Overall, this study shows that host mucosal niche and rearing environment are associated with distinct gut and gill microbiota patterns in L. crocea and provides preliminary ecological information for future microbiome-informed aquaculture management, functional validation of candidate taxa, and the development of land-based culture systems for marine fish.

Veterinary Sciences doi: 10.3390/vetsci13070709

Authors: Mengke Song Rongru Zhu Qian Zhang Meng Li Ming Tian Xinmiao He Xiuqin Yang

Skeletal muscle, accounting for about 40% of the body’s mass, is an important trait for farm animal production. Skeletal muscle satellite cells (MuSCs) are an excellent material for studying the regulatory mechanisms underlying muscle growth and regeneration after birth. Here, we analyzed the role and underlying mechanisms of Myc-associated zinc finger protein (MAZ) and secretory leukocyte protease inhibitor (SLPI) in porcine MuSC (pMuSC) proliferation for the first time. It was revealed that both MAZ and SLPI advanced the proliferation of pMuSCs by promoting cell cycle progression, and that a MAZ/SLPI axis regulated pMuSC proliferation. It was further shown that this MAZ/SLPI axis regulated pMuSC proliferation through the Wnt/β-catenin pathway. The proliferative role obtained above was confirmed in mouse myoblast C2C12 cells. Additionally, a single-nucleotide polymorphism (SNP), −360C>T, which frequently occurred in the promoter of the SLPI gene from Min, Yorkshire, and Longmin black pigs, was found to significantly increase the gene’s expression in vitro and in vivo. The SNP −360C>T in SLPI is a potential molecular marker predicting the proliferative ability of pMuSCs. The data provide a basis for controlling muscle growth and regeneration, which will provide benefits for improving the meat production of pigs and controlling muscle-related diseases in humans.

Veterinary Sciences doi: 10.3390/vetsci13070708

Authors: Aydin Daş Besime Doğan Daş Mücahit Kahraman Mehmet Avci Akın Yiğin Sadik Serkan Aydin Gülüzar Şengül Oğuz Ağyar Ahmet Yusuf Şengül

The aim of this study was to evaluate the effects of pomegranate syrup supplementation at levels of 0.0%, 0.1%, 0.2%, and 0.3% in the diets of quail (Coturnix coturnix japonica) on performance parameters, including body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), hot and cold carcass weights and carcass yields; meat quality characteristics, including L*, a*, b* color values, pH, and water-holding capacity (WHC) of breast and thigh meat; and oxidative stress parameters, including total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI); as well as glutathione peroxidase (GPx7) gene expression. A total of 120 newly hatched quail chicks were used in the experiment. The birds were assigned according to a completely randomized design with 15 replicates per treatment group and 2 birds per replicate and were randomly distributed into 60 cage compartments. While cumulative body weight gain (CBWG) was not significantly affected during the other experimental weeks (p > 0.05), statistically significant differences among the groups were observed only in the second week (p < 0.05). Feed intake values were not significantly different among the groups throughout all weeks of the experiment (p > 0.05). However, feed conversion ratio (FCR) was significantly different among the groups during all weeks (p < 0.05), with the pomegranate syrup-supplemented groups generally showing improved (lower) values compared with the control group. The inclusion of pomegranate syrup in the diets did not produce significant differences among the groups in breast meat quality parameters, including color values (L*, a*, b*), pH, and water-holding capacity. The obtained L*, a*, b*, and pH values were within the reference ranges reported in the literature. Evaluation of oxidative stress parameters demonstrated that pomegranate syrup supplementation increased TAS values, while significantly reducing TOS and OSI values (p < 0.05). Furthermore, GPx7 gene expression increased in the pomegranate syrup-supplemented groups compared with the control group, with the highest expression observed at the 0.1% supplementation level. In addition, toxicological evaluations have confirmed the safety of pomegranate preparations, further supporting their potential as feed additives.

Veterinary Sciences doi: 10.3390/vetsci13070707

Authors: Dany Elzahaby Bérénice Lutz Isabelle Iff

While colonoscopies are generally benign procedures, rare complications such as colonic perforation may result in life-threatening consequences. A 15-year-old neutered male British Shorthair cat underwent colonoscopy for investigation of chronic gastrointestinal signs and persistent tenesmus. During endoscopic evaluation of a distal colonic stricture, the cat developed sudden tachycardia, severe hypoxaemia, and a dramatic decline in end-tidal carbon dioxide (EtCO2), accompanied by diffuse subcutaneous emphysema. Intestinal perforation was suspected, and venous blood gas analysis revealed a markedly enlarged EtCO2–PvCO2 gradient, consistent with severe ventilation–perfusion mismatch and contributing to a presumptive diagnosis of pulmonary air embolism. Anaesthetic management focused on ventilatory adjustments, including an increased respiratory rate and initially high, then moderate peak inspiratory pressures, aimed at reducing EtCO2 and improving oxygenation. However, improvements in oxygen saturation were limited and the cat was euthanised. Contrast-enhanced post-mortem computed tomography confirmed a distal colonic perforation with extensive air dissemination, including pneumoretroperitoneum, pneumomediastinum, pneumothorax, and subcutaneous emphysema.

Veterinary Sciences doi: 10.3390/vetsci13070706

Authors: Andrei Răzvan Codea Alexandra Biriș Sidonia Gog-Bogdan Alina Diana Haşaş Daniela Neagu Cristian Popovici Mircea Mircean

Background: Feline hyperthyroidism is the most prevalent endocrine disorder of older cats; TT4 monitoring is costly and often inaccessible. Methods: Forty-three cats with primary hyperthyroidism were followed prospectively at baseline (T0) and at 1, 3 and 6 months (T1, T2, T3). Hct, Hgb, MCV, ALT and TT4 were measured at each visit. Per-timepoint correlations, repeated-measures correlation (rmcorr), linear mixed-effects models, and Friedman and ROC analyses were performed. Results: TT4 correlated with Hct (r = 0.875 at T1 to 0.943 at T3), MCV (0.907 to 0.964), Hgb (0.836 to 0.946) and ALT (0.919 to 0.981), intensifying progressively. Within individual cats, Hct (rmcorr r = 0.873) and MCV (rmcorr r = 0.881) tracked TT4, and for Hct, the within-cat coefficient exceeded the between-cat coefficient. All primary parameters and ALT changed significantly from T0 to T1 (all p < 0.001). ROC analysis identified Hct ≥ 35.2% at T2 as the optimal cut-off (AUC = 0.972, 95% CI 0.914–1.000; sensitivity 96.7%, specificity 92.3%). Conclusions: Hct and MCV tracked TT4 within individual cats during methimazole therapy and returned toward pre-treatment values after discontinuation. A rising Hct or MCV in a treated cat may signal loss of therapeutic control and prompt TT4 re-evaluation, without replacing hormonal monitoring.

Veterinary Sciences doi: 10.3390/vetsci13070705

Authors: Jorge Ramón López-Olvera

European wild ungulates are captured for different purposes, requiring the assessment of animal welfare and health. Specific guidelines to assess stress and pathophysiological compromise in captured wild ungulates are lacking. This review aims to set the bases for establishing standardised protocols allowing the evaluation of capture and handling stress in wild European ungulates. Physical and chemical capture and handling elicit stress, a physiological response activating sympathetic–adrenal medulla and hypothalamic–pituitary–adrenal cortex axes, releasing catecholamines and corticosteroids, respectively. While being an adaptive response, if prolonged over time, the effects become harmful and life-threatening, increasing body temperature and heart rate, and provoking muscular and renal ischemia. This clinical outcome is known as capture myopathy, which encompasses four sequential syndromes: Hyper acute or capture shock, Acute or ataxic-myoglobinuric, Sub-acute or ruptured muscle, and Chronic debility or delayed per-acute. Monitoring temperature, heart rate, serum muscular enzyme (CK, AST, ALT, and LDH) activities and lactate, creatinine, urea, and potassium concentrations characterises pathophysiological compromise. Wild ungulate capture and handling protocols should include not only methodology but also monitoring, data recording, sample collection, and analyses. This information should serve to improve protocols and eventually develop specific guidelines for stress and welfare assessment when capturing and handling wild ungulates.

Veterinary Sciences doi: 10.3390/vetsci13070703

Authors: Anna Antonella Spina Maria Elena Furfaro Vincenzo Chiofalo Alessandro Madeo Domenico Britti Cinzia Lucia Randazzo Georgiana Bosco Valeria Maria Morittu

Preserving the Apulo-Calabrese (AC), an endangered Italian autochthonous pig breed, requires sustainable nutritional strategies that improve pork quality without compromising productive performance. This study evaluated the effects of dietary supplementation with extruded linseed, alone or combined with grape pomace powder, rich in antioxidants, on growth performance and carcass traits in purebred Apulo-Calabrese (AC) pigs and Apulo-Calabrese × Goland (AC × G) crossbred pigs. Two independent feeding trials were conducted using three dietary treatments: a control diet (CTR), a diet containing 5% extruded linseed (SL), and a diet containing 5% extruded linseed plus 0.2% grape pomace powder rich in polyphenols (SLXI). Thirty-six purebred AC pigs reared outdoors, and 30 AC × G crossbred pigs reared indoors were included in the study. Body weight (BW) and average daily gain (ADG) were recorded throughout the trials, while hot carcass weight (HCW), carcass yield (CY), pH45, pH24, and backfat thickness (BF) were evaluated at slaughter. Data were analyzed separately for the two trials using Linear Mixed Models. Feed intake was complete in all groups, indicating good diet palatability. In purebred AC pigs, dietary treatment significantly affected overall ADG, with CTR pigs showing higher ADG than SLXI animals, while SL displayed intermediate values not significantly different from either CTR or SLXI. BW and carcass traits were not significantly influenced by diet. In addition, a significant Diet × Time interaction was detected for ADG (p = 0.010), indicating diet-dependent growth patterns over time. Sex did not significantly affect BW, ADG, or carcass traits in purebred pigs. In crossbred AC × G pigs, dietary treatments could not be evaluated inferentially due to the lack of pen-level replication; therefore, growth and carcass data are presented descriptively. However, Sex significantly affected carcass yield, with males showing higher CY than females (p = 0.004), while BW, ADG, and the remaining carcass traits were not influenced by Sex. The results suggest that extruded linseed, alone or combined with low-dose grape pomace powder, can be included in diets for purebred and crossbred pigs without compromising growth performance or basic carcass traits. These findings support the use of powdered grape pomacein feeding strategies aimed at the valorization and conservation of Apulo-Calabrese pig genetic resources. Further studies are needed to evaluate the effects of these dietary strategies on meat fatty acid composition, oxidative stability, sensory quality, and shelf life.

Veterinary Sciences doi: 10.3390/vetsci13070704

Authors: Weizhen Shen Mengran Zhang Yixiang Lian Jiahui Xu Yunye Jiang Jing Chen Jin Cui Bin Zhou

Porcine circovirus 3 (PCV3) is an emerging swine pathogen associated with reproductive disorders, multisystemic inflammation, and other clinical syndromes. The capsid protein (Cap), the principal structural and immunogenic component, is an ideal antigen for serological detection. However, its poor solubility in prokaryotic expression systems limits diagnostic assay development. In this study, the N-terminal 32 amino acids of Cap were truncated, and the protein was expressed in a soluble form using the SHuffle T7 Express strain. An indirect enzyme-linked immunosorbent assay (ELISA) using purified soluble Cap as a coating antigen was established and optimized. The assay exhibited excellent specificity, with no cross-reactivity against other common swine pathogens, high sensitivity, and good repeatability, with intra- and inter-assay coefficients of variation below 10%. The cut-off value was determined, and testing of 144 porcine serum samples showed a 78.47% concordance rate with PCR, indicating good diagnostic performance. This Cap-based indirect ELISA provides a reliable, cost-effective, and high-throughput tool for large-scale serological surveillance and epidemiological monitoring of PCV3, supporting improved disease control strategies in pig populations.

Veterinary Sciences doi: 10.3390/vetsci13070701

Authors: Xinhuai Shi Chaohao Wang Faxin Wen Kexin Zhang Shuna Liu Cong Zhu Changmin Hu Xinyue Chai

Limited data exist regarding how staphylococcal isolates from infected pets and their owners compare with domestic and international strains. In this study, swabs were collected from skin lesions of 160 pets and nasal cavities of 100 owners in Wuhan, China. Isolates were tested for antimicrobial susceptibility and the presence of resistance genes. Methicillin-resistant staphylococci (MRS, defined by the presence of mecA) were characterized using MLST, spa, and SCCmec typing. Among 268 staphylococcal isolates, 105 were coagulase-positive (69 S. pseudintermedius, 33 S. aureus, 3 S. schleiferi subsp. coagulans). S. pseudintermedius exhibited 91.3% susceptibility to oxacillin but 89.9% resistance to penicillin. Conversely, S. aureus was 97.0% susceptible to gentamicin and levofloxacin, yet 97.0% resistant to trimethoprim-sulfamethoxazole. Multidrug resistance was observed in 85.5% and 51.5% of these two species, respectively. Among the 26 mecA-positive isolates (21 MRSP, 5 MRSA), MLST analysis revealed household-level similarities in phenotypes and genotypes (e.g., ST25, ST22), suggesting potential transmission between pets and owners. Furthermore, these sequence types were closely related to globally prevalent lineages, indicating broader genetic backgrounds. Notably, immediate handwashing after pet contact reduced the likelihood of transmission (p = 0.04), especially when pets presented with dermatological disease. This study provides insights into infection control and resistance transfer.

Veterinary Sciences doi: 10.3390/vetsci13070702

Authors: Joanna Klećkowska-Nawrot Aleksandra Kroczak-Zdańkowska Adam Urantówka Grzegorz Zaniewicz Aleksander Chrószcz Dominik Poradowski

Woodpeckers (Picidae) are exposed to repeated mechanical loading and particulate material during climbing, drilling, pecking, and foraging on tree trunks, but the microscopic organization of their eyelids and nictitating membrane remains poorly known. This study asked whether European woodpeckers share a common histological pattern of ocular adnexa and whether selected structural features may be interpreted in the context of ocular surface protection. The upper eyelid, lower eyelid, and nictitating membrane were examined in 40 birds representing six European woodpecker species using routine histological and histochemical methods. In all of these species, the eyelids showed a layered organization with a keratinized anterior palpebral surface, non-keratinized conjunctival surface with goblet cells, marginal zone, musculofibrous stroma, and no tarsal glands. Species-related differences were observed in the examined material in epithelial thickness, skin fold development, melanin pigment distribution, and marginal zone organization. The lower eyelid contained a compact fibrous tarsal plate and CALT represented by lymphoid nodules and diffuse lymphocytes, with morphologically identified HEV-like venules located within or close to lymphoid tissue. The nictitating membrane showed regional variation in the marginal plait, leading edge, epithelial surfaces, stromal pigmentation, and fold organization. These findings provide the first comparative histological baseline for the eyelids and the nictitating membrane of European Picidae and support the cautious interpretation that these structures may contribute to mechanical, mucosal, and immune protection of the ocular surface in woodpeckers. Because sample sizes were unequal, these interspecific observations should be treated as descriptive, sample-level findings rather than definitive species-level traits.

Veterinary Sciences doi: 10.3390/vetsci13070700

Authors: Jinyan Meng Qinyao Wu Runlin Yu Zeyu Wen Sumeng Chen Yang Zhang Nuoyu Xu Shuyan Guo Xilu Sun Xingyuan Cao

Long-acting spot-on formulations are widely used for parasite control in cats, yet the pharmacokinetics of combined fluralaner and moxidectin remain incompletely defined beyond the recommended dose. This study evaluated the pharmacokinetics, absolute bioavailability, and dose proportionality of a novel fixed-combination spot-on formulation in healthy cats after topical administration of fluralaner/moxidectin at 40/2, 80/4, and 120/6 mg/kg BW, and compared intravenous administration of each compound alone or in combination. Forty-eight healthy cats were allocated to six groups of eight cats each. Non-compartmental analysis and a power model were used to characterize systemic exposure and dose proportionality. After intravenous co-administration, fluralaner exposure increased whereas moxidectin exposure decreased compared with single-agent administration, indicating a potential pharmacokinetic interaction. Topical administration produced slow absorption and sustained systemic persistence, with Tmax values occurring approximately 10–18 days after dosing and quantifiable concentrations maintained throughout the 160 days sampling period. Systemic exposure increased non-proportionally across the tested dose range, and absolute bioavailability was limited and dose-dependent, ranging from 9.7% to 26.9% for fluralaner and from 10.22% to 21.28% for moxidectin. No adverse reactions were observed. These findings demonstrate nonlinear topical absorption, and prolonged systemic persistence, supporting further dose optimization and formulation development of this long-acting feline antiparasitic delivery system.

Veterinary Sciences doi: 10.3390/vetsci13070699

Authors: Anyan Li Shimei Xu Ting Li Qiaoying Zeng Wei Wang

To develop an indirect ELISA kit for detecting serum antibodies against bovine herpesvirus 1 (BoHV-1), the gD protein of BoHV-1 was expressed in CHO-S cells and purified for use as the coating antigen. The optimized conditions were as follows: a coating antigen concentration of 2 µg/mL; serum dilution at 1:40 with incubation at 37 °C for 60 min; enzyme-labeled antibody dilution at 1:5000 with incubation at 37 °C for 60 min; and color development with TMB substrate at 37 °C for 15 min. The absorbance was then measured with a microplate reader. The kit could detect BoHV-1-positive serum at a dilution of up to 1:3200, showing favorable analytical detection capacity. No cross-reactivity was observed with positive sera against bovine viral diarrhea virus (BVDV), Akabane virus (AKAV), bovine parainfluenza virus type 3 (BPIV3), or lumpy skin disease virus (LSDV), reflecting ideal analytical specificity. Based on 126 field serum samples tested against a commercial IDEXX gB-based ELISA kit, we further calculated the diagnostic sensitivity (83.54%; 95% CI: 73.22–90.17%) and diagnostic specificity (100%; 95% CI: 92.43–100%) to quantitatively characterize its clinical detection performance. The ROC curve analysis based on IDEXX gB serostatus rather than true VNT-confirmed infection generated an AUC of 0.912 (95% CI: 0.856–0.951), which only reflects inter-assay concordance instead of absolute diagnostic validity. This indirect ELISA can be applied to epidemiological investigation and evaluation of immunization efficacy.

Veterinary Sciences doi: 10.3390/vetsci13070698

Authors: Timotei Pantea Sebastian Alexandru Popa Georgeta Stefan Ionica Iancu Vlad Iorgoni Alexandru Gligor Paula Nistor Janos Degi Luminita Costinar Corina Pascu Ionela Popa Alexandru Udrea David Purec Cosmin Horatiu Maris Viorel Herman

Q fever, caused by Coxiella burnetii, remains an important zoonotic disease with significant implications for animal health, livestock production, and public health. Epidemiological data regarding the circulation of C. burnetii in Romania remain limited, particularly in the western region of the country. This study aimed to assess the observed seropositivity and geographic distribution of anti-C. burnetii antibodies among sampled small ruminants from Western Romania. A cross-sectional sero-epidemiological survey was conducted using 546 serum samples collected from sheep (n = 451) and goats (n = 95) originating from five counties (Timiș, Arad, Bihor, Caraș-Severin, and Hunedoara). Sera were tested for IgG antibodies against C. burnetii using a commercial indirect enzyme-linked immunosorbent assay (ELISA). Overall, 87 animals were seropositive, corresponding to an observed seropositivity of 15.9% (87/546). Species-specific seropositivity was significantly higher in goats (25.3%) than in sheep (14.0%) (p = 0.006). Seropositive animals were detected in all surveyed counties, with prevalence ranging from 5.1% in Arad County to 21.3% in Timiș County. Significant geographic variation in seroprevalence was also observed among the investigated counties (p = 0.040). These findings indicate widespread serological evidence of exposure to Coxiella burnetii among the sampled small-ruminant population in Western Romania and reveal heterogeneous exposure patterns associated with both host species and geographic location. Continued surveillance integrating serological and molecular approaches is warranted to better characterize the epidemiology and transmission dynamics of C. burnetii within a One Health framework.

Veterinary Sciences doi: 10.3390/vetsci13070697

Authors: Jiawen Li Weidong Zhang Ximing Ren Jiarui He Leijun Wang Jujian Lv Kaihan Lin Wencai Du Rongjun Chen

This study aims to develop a lightweight, edge-deployable artificial intelligence (AI) system for real-time, non-contact recognition of cattle eating, standing, and lying behaviors in farm environments. Automated monitoring of these behaviors in cattle provides fundamental behavioral data for the future development of systems that analyze feeding duration, lying duration, and behavioral rhythms. Nevertheless, practical deployment on farms is hindered by data imbalance, dense animal groupings, scale variation, occlusion, and the need for low-cost edge computing. To address these challenges, we propose BoviFusionNet, a lightweight, edge-deployable AI system. A box balanced augmentation strategy rebalances training instances at the object level without altering the validation or test sets. Built upon YOLO11n, the model integrates three targeted enhancements: information-preserving downsampling (ADown), adaptive bidirectional feature fusion (BiFPN), and local window attention (C2CGA) to improve multi-scale representation and fine-grained behavior discrimination. Experimental results show that BoviFusionNet achieves 0.7851 recall, 0.7763 F1-score, 0.7976 mAP@0.50, and 0.6305 mAP@0.50:0.95, with only 5.4 GFLOPs and a 3.4 MB model size. Compared with the YOLO11n baseline, it improves mAP@0.50:0.95 by 9.92% and reduces the parameter count by 39.8%. After INT8 quantization and deployment on an RK3588S edge device, real-time inference reaches 28.08 frames per second (FPS). Therefore, BoviFusionNet offers an effective accuracy-complexity trade-off for on-farm edge AI applications. By enabling continuous, non-invasive monitoring of health-relevant behaviors, it provides fundamental behavioral data for the future development of veterinary health assessment tools without relying on cloud services or wearable sensors.

Veterinary Sciences doi: 10.3390/vetsci13070696

Authors: Adela Ioana Mustăţea Violeta Ștefania Rotărescu Adrian Gorbănescu Emilia Ciobotaru-Pîrvu

Animal abuse and neglect represent important challenges in companion-animal practice, with veterinarians playing a central role in their identification and management. This study investigated how Romanian companion-animal veterinarians recognise and manage suspected cases of abuse and neglect, and explored the relationships between professional training, familiarity with animal welfare legislation, and clinical practices. A cross-sectional survey was conducted using an original online questionnaire completed by 137 veterinarians. Descriptive statistics and inferential analyses were applied. Although 73% of respondents reported encountering suspected abuse cases, 93.4% said they had never received formal training in abuse identification, and 78.1% worked in practices without specific protocols for managing such cases. Reporting rates were low, with 71.5% of veterinarians never having submitted an official report. Neglect was identified as the most frequently encountered form of abuse. Significant associations were observed between familiarity with animal welfare legislation, level of training, and awareness of neglect, and veterinarians who perceived themselves as less prepared were more likely to express a need for additional education. These findings highlight critical gaps in veterinary training and institutional support, emphasising the need for structured educational programmes, lifelong learning and continuing professional development, standardised clinical procedures, and closer collaboration with authorities to improve animal welfare outcomes.

Veterinary Sciences doi: 10.3390/vetsci13070695

Authors: Karolina Śniegucka Maria Soroko-Dubrovina Paulina Zielińska Krzysztof D. Dudek

Dorsal metacarpal disease (DMD) is a common musculoskeletal injury in young racehorses, and effective non-invasive treatments remain of clinical interest. This study aimed to evaluate whether high-intensity laser therapy (HILT), which was applied as the sole intervention in horses retired from training, reduces the clinical signs of DMD compared with untreated controls. During the 2023–2024 racing seasons, 15 Arabian racehorses diagnosed with DMD were enrolled; 9 received HILT and 6 served as controls without laser therapy. The treatment protocol consisted of five daily HILT sessions followed by five sessions administered every other day. Thermographic, orthopedic, and radiographic examinations of the third metacarpal bones were performed before and after the treatment period in both groups, with additional thermographic and clinical assessments conducted throughout therapy. The results showed that horses treated with HILT did not exhibit a significant reduction in pain and lameness accompanied by thermographic changes consistent with decreased inflammation. These findings indicate that HILT did not alleviate clinical signs associated with DMD in the affected horses; however, further controlled studies are required to determine its effect on tissue healing processes and to optimize treatment duration.

Veterinary Sciences doi: 10.3390/vetsci13070694

Authors: Mohsen M. Alobre Ibrahim A. Alhidary Mohammed M. Qaid Abdulkareem M. Matar Hani H. Al-Baadani Abdulrahman S. Alharthi Ahmad A. Aboragah Ahmed A. Alghonaim Riyadh S. Aljumaah Mutassim M. Abdelrahman

(1) Background: Sunflower hulls (SFHs) are an abundant agro-industrial by-product that may serve as an alternative fiber source in sheep diets, particularly in arid production systems. This study evaluated the effects of graded levels of dietary SFH during late gestation and lactation on colostrum and milk composition, lamb growth performance, neonatal survival, and serum biochemical parameters in Naemi sheep. (2) Methods: Seventy-two primiparous Naemi ewes (initial body weight = 55.0 ± 3.8 kg) were randomly allocated to four dietary treatments containing 0, 12, 20, or 28% SFH (n = 18 ewes per treatment) from late gestation until lamb weaning at 90 days postpartum. Data were analyzed using mixed-model procedures, and treatment effects were considered significant at p ≤ 0.05. (3) Results: Dietary SFH supplementation did not affect lamb birth weight or overall survival rate (p > 0.05). However, weaning weight declined from 28.03 kg in the control group to 25.11 kg in the 20% SFH group, showing a significant linear decrease with increasing SFH inclusion (p = 0.044). Likewise, average daily gain decreased linearly by 10.4% (p = 0.044). Colostrum composition was partially affected by dietary treatment, with fat and protein concentrations at 48 h postpartum increasing quadratically by 46.6% and 33.50%, respectively, in ewes fed 20% SFH compared with the control group (p < 0.05). In contrast, milk fat and total solids contents decreased quadratically by 30.7% and 13.4%, respectively, in ewes receiving 12% SFH (p = 0.01). Most serum metabolite variables were unaffected by dietary treatment and remained within normal physiological reference ranges. However, overall serum triglyceride concentration decreased linearly with increasing SFH inclusion (p = 0.023). (4) In conclusion, SFH can be incorporated into the diets of Naemi ewes during late gestation and lactation as an alternative fiber source without adversely affecting lamb birth weight, survival, or overall metabolic health. However, increasing dietary SFH inclusion reduced pre-weaning lamb growth despite improvements in selected colostrum quality traits. Based on the productive and physiological responses observed, 12% dietary SFH appears to be the optimal inclusion level, providing the best balance between animal performance and sustainable feed utilization.

Veterinary Sciences doi: 10.3390/vetsci13070693

Authors: Tong Mu Zhixuan Qiao Honghong Hu Yun Ma Ziyan Jiang Yuxin Huang Zhihong Sun

Lactation traits are important indicators for evaluating the production performance of dairy cows and the economic efficiency of dairy production. Identifying potential regulatory genes is essential for elucidating the molecular mechanisms underlying lactation and facilitating molecular breeding. This study aimed to identify candidate genes and regulatory pathways potentially associated with lactation traits in dairy cows by integrating transcriptome expression profiles of primary bovine mammary epithelial cells (BMECs) from eight Holstein cows with lactation phenotypic data. A gene co-expression network was constructed using weighted gene co-expression network analysis (WGCNA). Co-expression modules associated with lactation traits were identified, and candidate genes were further screened by integrating gene significance, module membership, functional enrichment analysis, random forest analysis, gene-phenotype association analysis, single-gene gene set enrichment analysis (GSEA), ROC curve analysis, and tissue expression profiling. Four co-expression modules, namely MEdarkturquoise, MEsteelblue, MEbrown, and MEskyblue3, were significantly associated with lactation traits (p < 0.05). Genes in these modules were mainly enriched in biological processes and pathways related to ribosome biogenesis, protein translation, the cell cycle, oxidative phosphorylation, and the PI3K-Akt signaling pathway, which may be involved in lactation regulation. Through multi-strategy cross-screening, four candidate genes were ultimately identified. MDM2 was associated with daily milk yield (DYM), RPL37A with total milk solids (TMS), and SLC25A25 and CCNE2 with milk fat percentage (MFP) and fat-to-protein percentage ratio (FPP). Reverse transcription quantitative real-time PCR (RT-qPCR) validation showed that MDM2 and RPL37A were relatively highly expressed in mammary tissue, suggesting that they may be involved in lactation-related biological processes in dairy cows. These findings provide potential candidate regulators and theoretical support for further studies on the molecular mechanisms of lactation traits; however, their functional roles require further validation through in vitro and in vivo experiments.

Veterinary Sciences doi: 10.3390/vetsci13070691

Authors: Alessandro Spadari Federica Meistro Raimondo Tornago Cristian Facchetti Paola D’Angelo Maria Virginia Ralletti Salvatore Montemagno Riccardo Rinnovati

Despite the wide range of therapeutic options currently available for equine sarcoids, recurrence of these lesions remains a major clinical challenge. Consequently, the identification of effective strategies for the management of recurrent sarcoids remains an important objective in equine oncology. The aim of this retrospective study was to evaluate the clinical outcome of surgical cytoreduction combined with intraoperative electrochemotherapy in horses affected by recurrent equine sarcoids. Medical records of horses treated between July 2018 and August 2025 at the Veterinary Teaching Hospital of the University of Bologna were retrospectively reviewed. Data collected included signalment, lesion location, clinical subtype, lesion size, chemotherapeutic agent used, recurrence status, and follow-up duration. Fourteen horses presenting 23 recurrent sarcoid lesions met the inclusion criteria. Recurrence was evaluated at both the lesion and horse levels. Local tumour control was achieved in 16 of 23 lesions (69.6%). Recurrence was observed in 7 lesions, corresponding to a lesion-based recurrence rate of 30.4%, and in 5 of 14 horses (35.7%). Malignant and fibroblastic lesions showed higher recurrence rates than other clinical subtypes, whereas larger lesions tended to recur more frequently than smaller lesions. No significant differences were identified between cisplatin- and carboplatin-treated lesions. Surgical cytoreduction combined with intraoperative electrochemotherapy provided encouraging local tumour control in a population composed exclusively of recurrent sarcoids. This treatment protocol may represent a valuable therapeutic option for horses with local recurrence of sarcoids, although larger prospective studies are required to further define prognostic factors and long-term outcomes.

Veterinary Sciences doi: 10.3390/vetsci13070692

Authors: Mingfa Yang Wei Yu Xiaofang Chen Wei Zhang Qingwen Meng Changwen Li Jiasen Liu Changyou Xia

Duck hepatitis B virus (DHBV), Duck adenovirus 3 (DAdV-3) and Streptococcus gallolyticus subsp. pasteurianus (SGSP) are three distinct pathogens associated with infectious diseases in waterfowl. Clinical differentiation among these agents is often difficult due to overlapping signs, underscoring the need for accurate, rapid, and simultaneous diagnostic tools to support early detection and effective disease surveillance. In this study, we developed and validated a triplex TaqMan real-time quantitative PCR (qPCR) assay for the simultaneous detection and quantification of DHBV, DAdV-3, and SGSP. The assay indicated high analytical sensitivity, achieving detection limits of 10 copies/µL for each target, with all standard curves showing excellent linearity (R2 = 0.999). The evaluation of specificity showed that the triplex qPCR assay had high specificity for the three target pathogens, as it did not yield any cross-reactive signals with other pathogens examined. Intra-assay and inter-assay coefficients of variation for recombinant plasmid standards were consistently <2%, indicating high reproducibility. Meanwhile, the assay was employed to investigate the epidemiology of the three pathogens in ducks. The positive rates for DHBV, DAdV-3 and SGSP were 73.95% (159/215), 11.16% (24/215) and 37.21% (80/215), respectively. The co-infection rates were 5.58% (12/215) for DHBV and DAdV-3, 27.91% (60/215) for DHBV and SGSP, 2.33% (5/215) for DAdV-3 and SGSP, and 2.33% (5/215) for all three pathogens. In summary, this triplex qPCR assay displays excellent specificity, sensitivity and repeatability, which may serve as a reliable testing tool for routine clinical diagnosis, epidemiological monitoring and co-infection screening of the three target pathogens in locally sampled duck populations.

Veterinary Sciences doi: 10.3390/vetsci13070690

Authors: Carmen Davias Alejandro Perez-Ecija Daniela Ruiz-Lastra Adelaida De Las Heras Ramiro Toribio Francisco J. Mendoza

The Immulite 2000xpi, a chemiluminescent immunoassay (CLIA)-based analyzer, is currently recommended for measuring thyroid hormones (TH) concentrations in equids. However, scarce data are available on TH measurements in donkeys using this technique. Similarly, the effects of factors such as age and gender on TH concentrations have not been assessed with this analyzer. Serum free and total T4 and T3 concentrations were measured in 40 healthy donkeys (6.1 ± 0.8 years old; 8 intact jacks and 32 jennies) and 41 horses (10.7 ± 0.5 years old; 15 stallions and 26 geldings) using the Immulite 2000xpi. A Mann–Whitney test was used to compare two groups, and a Kruskal–Wallis test followed by Dunn’s post hoc test was used for comparisons among more than two groups. This analyzer had lower intra-assay variability and fewer measurements under the detection limit for tT4 and fT4 than for tT3 and fT3 determinations. The canine TSH kit for thyroid-stimulating hormone (TSH) showed high imprecision. Donkeys had higher serum TH concentrations than horses. No differences were observed among gender or age groups. This is the first study to report TH concentrations in donkeys using a CLIA-based analyzer and to evaluate the effects of several factors on TH concentrations. The canine TSH kit is not valid for measuring TSH in equids.

Veterinary Sciences doi: 10.3390/vetsci13070689

Authors: Mônica Duarte da Silva Christina Ramires Ferreira Hianka Jasmyne Costa de Carvalho Sandra Maria Barbalho Rodrigo da Silva Nunes Barreto Rosa Direito Maria Angélica Miglino

Canine mixed-type mammary carcinomas (CMTs) exhibit remarkable histological and structural heterogeneity, sharing some biological similarities with human breast cancer and supporting their relevance as models in comparative oncology. These tumors frequently contain regions of malignant epithelial cells, areas of mesenchymal differentiation, and ossified tissue, reflecting their complex histological organization. In this exploratory pilot study, we performed MRM-based spatial lipidomic analysis of distinct tumor regions; namely the margin, middle (epithelial/myoepithelial-rich), and central ossified areas. Our region-resolved lipidomic analysis revealed distinct lipidomic signatures across regions, with significant alterations in phosphatidylcholines, phosphatidylethanolamines, sphingomyelins, triacylglycerides, and free fatty acids distribution. The central ossified area showed enrichment in structural lipids and ceramides, while the tumor margin demonstrated enrichment of ether lipids, which may reflect differences in tissue composition and local metabolic environments. By characterizing region-specific lipid profiles, this study provides insights into metabolic heterogeneity within a canine mixed mammary carcinoma. These findings highlight the potential of spatial lipidomics for investigating intratumoral metabolic heterogeneity and provide preliminary data to support future studies in comparative oncology involving canine and human mammary tumors.

Veterinary Sciences doi: 10.3390/vetsci13070688

Authors: Yingni Sun Shuyun Xu Zhijie Luo Taishan Fan Xia Zhou

To investigate the biological characteristics of microbes in fresh milk, this study gathered fresh milk from seven large-scale dairy farms in the southern and northern regions of Xinjiang, analyzing the composition and abundance of bacterial communities in these samples through metagenomic sequencing technology. Firmicutes, Proteobacteria, and Actinobacteria were consistently identified as the dominant phyla across all samples, with stable relative abundance patterns across regions. At the genus level, the genera with the highest relative abundances were Sporosarcina, Streptococcus, and Escherichia, with relative abundances of 2.51–2.58%, 2.23–2.27%, and 1.93–1.97%, respectively. While exploring species richness, it was observed that the XN group had the most OTUs, the DR group had the fewest, and there were significant differences in community structure between the ND group and the other six groups. Further Alpha diversity analysis revealed no significant variation in Chao1 indices across the seven sample groups, highlighting a significant difference in Shannon index for ND samples, and no significant differences in Shannon indices between the CJ, JY, KT, DR, and TR samples. Shifting focus to functional potential, the top three relative abundances in the microbial metagenome KEGG functional library are biological systems, human diseases, and environmental information processing; additionally, within the CAZy (Carbohydrate-Active enZymes) database, the three most abundant categories are glycosyltransferases (GT), glycoside hydrolases (GH), and carbohydrate-binding modules (CBM). By delineating these patterns, this study demonstrates the microbial spectrum characteristics of fresh milk from southern and northern Xinjiang, China, offering a theoretical foundation for enhancing the quality of fresh milk in the area.

Veterinary Sciences doi: 10.3390/vetsci13070687

Authors: Bakhit Baimenov Kalamkas Malikzada Alma Dossova Zhanaidar Bermukhametov Pavel Shevchenko Albina Gabitova Oxana Tomaruk Aliya Yskak Gulzhan Chuzhebayeva Raushan Rychshanova

Staphylococcus aureus is a leading cause of foodborne illness in animal-derived food products, yet comprehensive data on its antimicrobial resistance and toxigenic potential from Central Asia remain scarce. We examined 711 food samples (meat, milk, and dairy products) from the Kostanay region of Kazakhstan (2024–2025). Isolates were identified by MALDI-TOF mass spectrometry; antimicrobial susceptibility was assessed by disk diffusion (EUCAST v.16.0); enterotoxin production (types A–E) by ELISA; and resistance and toxin genes by PCR. Concordance between methods was evaluated using Cohen’s kappa coefficient. S. aureus was recovered from 11.8% of samples (84/711), with raw cow’s milk carrying the highest contamination risk (OR = 2.75; 95% CI: 1.74–4.38). High resistance to ampicillin (52.4%) and benzylpenicillin (50.0%) was found; 20.2% of strains were multidrug-resistant. Among the 17 cefoxitin-resistant isolates, only 4 harbored mecA, suggesting that 13 isolates (15.5%) should be interpreted as cefoxitin-resistant, mecA-negative isolates with a putative BORSA-like phenotype rather than as confirmed MRSA. Staphylococcal enterotoxins A–E were detected in 26.2% of isolates, and egc-cluster genes seg and sei were present in 9.5% each. The strongest genotype–phenotype concordance was observed for aph(3′)/kanamycin (κ = 0.65) and blaZ/benzylpenicillin (κ = 0.64). These results underscore the need for combined molecular and phenotypic surveillance for regional food safety monitoring.

Veterinary Sciences doi: 10.3390/vetsci13070686

Authors: Maria Virginia Ralletti Andrea Beltrame Elisa Marcucci Paola D’Angelo Riccardo Rinnovati Alessandro Spadari

Tibial fractures in beef cattle are a significant clinical challenge, particularly in animals weighing over 300 kg, where surgical treatment is often discouraged due to poor prognosis. This study evaluated the use of full limb double transfixation pin casting (TPC) in field settings for treating tibial fractures in 11 beef cattle weighing between 260 and 480 kg. All animals received the same surgical approach, involving pin placement both proximal and distal to the fracture site, followed by application of a fiberglass cast encompassing the entire limb. The short-term outcome was positive in 90.9% of cases, with two animals not surviving to cast removal. A long-term positive outcome—defined as completing the production cycle and reaching slaughter weight—was achieved in 72.7% of cases. All surviving animals exhibited some residual lameness, but none experienced functional limitations or carcass devaluation. These results support the feasibility and effectiveness of a full limb cast with double transfixation pinning for tibial fracture repair in heavy cattle under field conditions, offering a cost-effective and practical alternative to internal fixation or Thomas splint casts. Further studies are encouraged to confirm these findings and better assess outcomes in open fracture cases.

Veterinary Sciences doi: 10.3390/vetsci13070685

Authors: Jianliang Zhang Penghao Wei Yidan Wang Zhiyan Sun Junjun He Feiyan Dai Jianfa Yang Fengcai Zou Luyang Wang

Migratory birds are potential carriers for the cross-regional spread of zoonotic diseases due to their high mobility and extensive geographical activity range. Blastocystis, a ubiquitous parasitic protist, can infect a broad spectrum of animal hosts including humans, constituting a potential public health hazard. Currently, data on Blastocystis infection in black-headed gulls (Chroicocephalus ridibundus) remain scarce. This study aimed to investigate the fecal sample positivity rate, epidemiological characteristics, and subtype distribution of Blastocystis in wintering black-headed gulls in Kunming, China. A total of 245 fresh fecal samples were collected from black-headed gulls in Kunming, Yunnan Province, during December 2025 and January 2026. Fecal DNA was extracted and subjected to PCR amplification targeting the small subunit ribosomal RNA (SSU rRNA) gene of Blastocystis. Results showed a 3.27% (8/245) overall Blastocystis fecal sample positivity rate in wintering black-headed gulls, with two subtypes detected: dominant zoonotic pathogenic ST7 (87.5%, 7/8) and ruminant-adapted ST26 (12.5%, 1/8). This study represents the first report of Blastocystis in black-headed gulls in China, providing data supporting the understanding of genetic subtype diversity of Blastocystis in avian hosts.

Veterinary Sciences doi: 10.3390/vetsci13070684

Authors: Shiyu Zhang Sheng Lu Jianan Liu Zhonghao Chen Caiying Li Jiale Ma Min Sun Xinming Pan

Porcine diarrheagenic Escherichia coli strains exhibit heterogeneous virulence profiles involving multiple factors, complicating broad-spectrum serological diagnosis. Here, we designed a multiepitope fusion antigen, MEAET, containing predicted B-cell epitopes from six major virulence factors, K88/F4, F18, LT, Stx2e, HlyA, and Tir, and developed an indirect ELISA for broad antibody detection. Structural modeling predicted a well-folded conformation with multiple discontinuous B-cell epitopes, and molecular docking suggested favorable interactions between the C-terminal dendritic cell-targeting peptide and the porcine immune receptor SLA1. Immunization of piglets with purified recombinant MEAET generated hyperimmune serum with a titer exceeding 1:102,400, which specifically recognized all six individual antigens by Western blot. The optimized ELISA demonstrated satisfactory repeatability (intra-assay CV ≤ 4.26%) and reproducibility (inter-assay CV ≤ 5.57%), showed no detectable cross-reactivity with antibodies against other tested porcine pathogens, and exhibited good analytical sensitivity, with positive sera detectable at dilutions up to 1:3200. The optimized ELISA showed good repeatability and reproducibility, with intra-assay and inter-assay coefficients of variation of ≤4.26% and ≤5.57%, respectively, no detectable cross-reactivity with the tested heterologous controls, and good analytical sensitivity, with positive sera remaining detectable at dilutions up to 1:3200. The S/P ratio cut-off was 0.203. In field sera, positivity rates were 74.5% (35/47) in diarrheic pigs and 34.0% (36/106) in asymptomatic pigs. These primary results indicate that the MEAET-based indirect ELISA represents a promising tool for serological detection of antibodies against multiple virulence factors of diarrheagenic E. coli in pigs and for field surveillance in swine herds.

Veterinary Sciences doi: 10.3390/vetsci13070683

Authors: Dongyu Huang Lu Zhang Chunyu Xue Leimin Zhang Mingchun Ren Xiaodi Xu Hualiang Liang Haifeng Mi

An 8-week trial evaluated dietary copper methionine (CM) on body and flesh color in yellowed channel catfish. Basic diets were supplemented with CM at 0 (control, Cu1), 7 (Cu2), 14 (Cu3), 21 (Cu4), and 27 (Cu5) mg/kg, with corresponding measured copper levels of 10.82, 12.63, 14.35, 15.58, and 17.40 mg/kg. In skin, GSH levels in Cu2-Cu5 groups and GSH-Px activities in Cu2-Cu4 groups were increased, and in muscle, CAT activities in Cu3-Cu5 groups and GSH levels in Cu4-Cu5 groups were increased. Gene expression analysis showed downregulated keap1 (Cu2–Cu4 in skin; Cu3–Cu5 in muscle) and nfκb (Cu2–Cu5 in skin), upregulated nrf2 (Cu3–Cu5 in skin; Cu4 in muscle) and il-10 (Cu4 in skin). Colorimetric analysis showed reduced the values of b* in skin (Cu3–Cu5) and muscle (Cu2–Cu5). Uranidin content decreased in skin (Cu3–Cu5) and muscle (Cu4–Cu5), while melanin and tyrosinase increased in skin (melanin: Cu2–Cu3; tyrosinase: Cu3–Cu4) and muscle (Cu3–Cu5). The expression of melanogenesis-related genes (mitf, tyr, etb, camk2, slc24a5) was increased in skin and muscle of Cu3-Cu5 groups, while the uranidin synthesis gene xdh was downregulated. Overall, optimal effects were observed at 14–21 mg/kg CM (measured copper: 14.35–15.58 mg/kg).

Veterinary Sciences doi: 10.3390/vetsci13070682

Authors: Jiankun Pang Qipeng Zhang Chen Lv Huawei Li Xuyong Zhao Ruining Wang Keshan Zhang Yaqiong Ye Mengmeng Zhao

Porcine reproductive and respiratory syndrome (PRRS) has been prevalent in Germany for over 30 years, posing a significant threat to the local swine industry. There are limited analyses to offer German PRRSV ORF5 genetic and evolutionary characteristics and GP5 structural and functional features. In this study, a total of 518 sequences of the PRRSV GP5 gene were obtained from the GenBank database, encompassing 102 sequences from Germany to investigate the genetic relationships and GP5 structural and functional features. Similarly, phylogenetic and recombination analyses were used to identify genetic relationships. Two German PRRSV-1 lineage 1 strains exhibited more than 98.0% nucleotide similarity with vaccine strains. Eighteen German PRRSV-2 lineage 5 strains exhibited more than 98.0% nucleotide similarity with a prototype vaccine strain. German PRRSV-1 lineage 1 was predominant. GP5 structural and functional features were determined by N-glycosylation sites, B-cell epitopes, and transmembrane domain predictions. The main N-glycosylation patterns and transmembrane domains of German PRRSV-1 lineage 1 were time-dependent. Mutations of German PRRSV-1 lineage 3 in the primary neutralizing epitope were detected. In conclusion, these findings reveal lineage-specific molecular variation and improve the understanding of the molecular evolution of German PRRSV.

Veterinary Sciences doi: 10.3390/vetsci13070681

Authors: Jihee Han Sijin Cha Jeonghyun Seo Kunho Song

Synchronous low-grade alimentary lymphoma and gastrointestinal mast cell tumor have recently been described in cats. This report describes synchronous low-grade alimentary lymphoma and gastrointestinal mast cell tumor involving the jejunum in a cat. A 2.3 kg, 15-year-old, spayed female Chinchilla Persian cat presented with vomiting, diarrhea, anorexia, and lethargy. Ultrasonography and computed tomography demonstrated diffuse intestinal wall thickening and a focal jejunal lesion characterized by marked transmural thickening with loss of wall layering. Exploratory laparotomy and full-thickness intestinal biopsies were conducted. Histopathologic and immunohistochemical examination revealed two morphologically distinct neoplastic populations involving different jejunal wall layers. The two populations were anatomically adjacent but remained distinct without convincing intermingling in the examined histologic sections. These findings supported the interpretation of a lesion most consistent with a collision pattern. The affected intestinal segment was surgically resected, and chlorambucil and prednisolone were administered postoperatively. Clinical signs resolved, imaging abnormalities improved, and no evidence of local gastrointestinal mast cell tumor recurrence or progression of low-grade alimentary lymphoma was detected during the 14-month follow-up period. This case expands the currently recognized morphologic spectrum of synchronous feline intestinal neoplasia and highlights the importance of comprehensive histopathologic assessment and full-thickness intestinal biopsies for accurate diagnosis.

Veterinary Sciences doi: 10.3390/vetsci13070680

Authors: Sin-Yeong Park Youjung Jung Dongbin Lee Chang-Hwan Moon

The middle hepatic vein (MHV) may lie in close proximity to the gallbladder fossa and represent a potential source of severe hemorrhage during cholecystectomy. While this anatomical relationship is well recognized in human surgery, the clinical relevance of MHV proximity during canine cholecystectomy has rarely been described. Two dogs underwent cholecystectomy for gallbladder disease. In Case 1, an 8-year-old Pomeranian with gallbladder mucocele and necrotizing cholecystitis developed sudden massive hemorrhage during dissection of the gallbladder from the hepatic fossa between the quadrate and right medial liver lobes, when a branch of the middle hepatic vein (MHV) adherent to the gallbladder wall ruptured, causing rapid hypotension (mean arterial pressure 10–20 mmHg). Hemostasis was achieved using hemoclips, and the dog recovered following transfusion. In Case 2, an 11-year-old Spitz with a gallbladder mass underwent cholecystectomy and liver biopsy. Preoperative computed tomography demonstrated MHV branches coursing within <1 mm of the gallbladder wall. Careful dissection preserved the venous structures, and surgery was completed without significant hemorrhage. MHV branches may occur immediately adjacent to the gallbladder fossa in dogs and may represent an underrecognized source of hemorrhage during cholecystectomy. Awareness of this relationship and preoperative vascular evaluation in selected cases may improve surgical safety.

Veterinary Sciences doi: 10.3390/vetsci13070679

Authors: Kehamo Abi Zihan Xia Lanmuyi Gou Wentao Zhang Kegu Ji’e Shenglin Li Taichun Gao Wangqing Banma Falong Yang

Sheep are a major livestock species in China, yet pneumonia-related mortality poses a significant obstacle to intensive farming. In this study, 115 sheep lung samples were collected and classified into different pneumonia severity groups based on lung lesion scoring. Subsequently, this study employed 16S rRNA sequencing to systematically investigate the structure and diversity of the pulmonary microbiota in sheep, including alpha diversity, beta diversity, and LEfSe analyses. Metagenomic techniques were also applied to analyze the abundance of metabolic pathways, exploring the associations between functional gene differences and pneumonia severity, as well as putative antibiotic resistance genes, virulence factors, and the species contributions of functional genes in severe pneumonia cases. Microbial richness and diversity were significantly higher in the severe pneumonia group than in the healthy/mild lesion group (p < 0.05). While the dominant microbial structures were similar across the groups, notable differences were observed in the abundance of respiratory disease-associated genera, with Pasteurella, Mannheimia, Mycoplasma, Bibersteinia, and Moraxella identified as significantly enriched in severe cases. Moreover, several genera originating from the gut and oral cavity were also associated with pneumonia, suggesting a potential gut–lung axis. Carbohydrate metabolism was the most prevalent pathway in all groups, whereas amino acid metabolism was significantly enriched in the severe pneumonia group. Putative antibiotic resistance genes were differentially enriched; the severe pneumonia group showed significant enrichment of genes conferring resistance to aminoglycosides, tetracyclines, and polymyxins. Virulence factor analysis identified nutritional/metabolic factors and adhesion as the predominant virulence mechanisms. Species contribution analysis further revealed that Mannheimia, Mycoplasma, Pasteurella, and Moraxella were the predominant species associated with functional gene enrichment. In conclusion, the current study reveals associations between changes in the pulmonary microbiota structure and function and the severity of pneumonia in sheep, aiming to provide a foundation for future hypothesis-driven research on the role of the pulmonary microbiota in pneumonia progression.

Veterinary Sciences doi: 10.3390/vetsci13070678

Authors: Guang Yang Pengyun Ji Lu Zhang Zhou Yu Guoshi Liu

Heat stress impairs male reproductive function, but whether acute heat stress alters glutamine-derived carbon labeling in Sertoli cells and whether melatonin modulates this response remain unclear. This study aimed to determine whether acute heat stress alters glutamine-derived carbon labeling in primary goat Sertoli cells and to evaluate whether melatonin attenuates these heat stress-associated metabolic changes. Primary goat Sertoli cells were assigned to control, heat-stress, or heat-stress plus melatonin groups. Cells were labeled for 24 h with [U-13C5] glutamine. Cells in the heat-stress and heat-stress plus melatonin groups were subsequently exposed to 42 °C for 0.5 h, whereas control cells were maintained at 37 °C. In the heat-stress plus melatonin group, melatonin (0.5 μM) was applied throughout labeling and heat exposure. GC–MS was used to measure mass isotopologue distributions, which were subsequently corrected for natural isotope abundance, and to calculate total 13C-labeled fractions of selected TCA cycle intermediates; extracellular acidification rate (ECAR)-derived parameters were measured as indirect indices of glycolysis-associated acidification. Heat stress increased the total 13C-labeled fraction of cis-aconitate and, as an exploratory combined readout, the unweighted average total 13C-labeled fraction of citrate and cis-aconitate; succinate and malate were unchanged, while fumarate decreased. Melatonin reduced the heat stress-associated total 13C-labeled fractions of citrate and cis-aconitate and decreased the relative abundance of cis-aconitate M + 5. Melatonin also attenuated heat stress-associated increases in ECAR-derived glycolysis, glycolytic capacity, and glycolytic reserve. These findings indicate that acute heat stress is associated with altered citrate/cis-aconitate 13C-labeling patterns that are potentially compatible with reductive carboxylation-related labeling but do not directly demonstrate altered pathway flux; these changes were attenuated by melatonin. Overall, the study objective was achieved by showing that acute heat stress altered citrate/cis-aconitate 13C-labeling patterns and increased ECAR-derived extracellular acidification in primary goat Sertoli cells, and that melatonin attenuated both responses. Further studies incorporating a melatonin-only group, dynamic isotope tracing, mitochondrial respiration measurements, redox analysis, and flux modeling are warranted to define the underlying metabolic routes more directly.

Veterinary Sciences doi: 10.3390/vetsci13070677

Authors: Tyler Niblett Kimberly Williams Marguerite Cross Alexis Clapp Mark Estienne

Previous work demonstrates that short -term increases in feed allowances (i.e., “nutritional flushing”) can improve ovulation rate. Furthermore, precision livestock farming equipment like electronic sow feeder (ESF) systems are becoming more commonplace to enhance individual sow management in group housing. The objective of this study was to determine the effects of short-term increases in feed allowances on growth and sexual development in previously feed-restricted gilts using an ESF. Prepubertal crossbred gilts were acclimated to an ESF and allocated to one of three feeding regimens, ad libitum (AD, n = 14), restricted (RS, n = 14; 2.72 kg/d), or 2.72 kg/d, until 7 days after puberty (NF, n = 14), followed by ad libitum feeding. Estrus detection and feeding regimens commenced at 160 days of age and continued for 48 days. Reproductive tracts were harvested between 8 and 11 days after second estrus. Average daily gain was improved in NF gilts compared to RS gilts (p = 0.04) Ad libitum-fed gilts were less feed efficient than the other treatment groups (p < 0.01). Improved ovulation rate was observed for NF gilts compared to RS gilts (p = 0.05). Feed disappearance decreased as estrus approached for AD and NF gilts (p < 0.01). For the regimens herein, age at puberty onset was not impacted; however, feed conversion was improved for the level of restriction implemented. Moreover, employing ESF technology may support current estrus detection methods by monitoring feeding behavior for AD-fed gilts.

Veterinary Sciences doi: 10.3390/vetsci13070676

Authors: Renato Leon Amanda Blubaugh Haley Starr Frane Banovic

Intradermal injections of anti-canine immunoglobulin E (IgE) in healthy dogs have been utilized in preclinical drug testing to evaluate the efficacy of anti-allergic drugs used to treat canine atopic dermatitis (AD). However, the molecular effects of established canine anti-allergic drugs on this acute canine IgE-mediated atopic model remain largely uninvestigated. The objective of this study was to characterize the effect of proactive oclacitinib and prednisolone treatments on the immune and skin barrier transcriptome of IgE-mediated late-phase reactions (LPRs) in an acute model of canine AD. Sixteen healthy adult research-bred beagles were randomized to receive either oclacitinib or prednisolone orally for six days, followed by an intradermal anti-canine IgE injection. Biopsies were collected 24 h post-injection for RNA isolation and sequencing; previously analyzed transcriptomes (healthy skin, saline-injected skin, IgE lesions without drug modulation) from the same colony of dogs served as controls. Administration of prednisolone and oclacitinib prior to intradermal anti-IgE injections reduced the number of differentially expressed genes (DEGs) in 24 h samples to 1251 and 1471, respectively. Both treatments resulted in a decrease in expression of several significantly upregulated T helper-(Th)1 (e.g., MX1, OAS1, STAT1), Th2 (e.g., CCL13, CCL8, IL13RA1, IL-33, IL5RA, OSM), chemokine and receptor (e.g., CCL19, CCL2, CCL3, CCR1, CCR3) genes in comparison to the untreated IgE-mediated lesions. Interestingly, only prednisolone treatment significantly reduced IL-13 upregulation, an important gene in the Th2 immune response. In conclusion, both prednisolone and oclacitinib reduced the transcriptomic changes observed in the acute lesions of the canine IgE-induced atopic dermatitis model, with prednisolone inducing a broader inhibitory immune response.

Veterinary Sciences doi: 10.3390/vetsci13070675

Authors: Nisachon Apinda Wasana Chaisri Terdsak Yano Suwit Chotinan Thanaporn Eiamsam-ang Saruda Tiwananthagorn Panuwat Yamsakul

Coccidiosis is one of the most economically important parasitic diseases in poultry, causing intestinal damage, impaired nutrient utilization, and substantial production losses. The extensive use of anticoccidial drugs has raised concerns regarding drug resistance, chemical residues in poultry products, and the environmental burden associated with veterinary drug use, highlighting the need for complementary control strategies. This study evaluated the anticoccidial efficacy of phytogenic formulations containing Curcuma longa and Piper longum extracts prepared using a solid-dispersion technique designed to enhance the dissolution characteristics of poorly water-soluble phytogenic compounds. The in vitro activity of the formulations was evaluated against mixed Eimeria spp. field-isolated oocysts using scanning electron microscopy (SEM) to examine ultrastructural alterations, whereas in vivo efficacy was investigated in broiler chickens experimentally challenged with Eimeria tenella. Cecal lesion scores, oocyst shedding, and growth performance were evaluated as indicators of infection severity and treatment response. SEM observations revealed marked structural damage to mixed Eimeria spp. oocysts following exposure to the phytogenic formulations. In the animal trial, all phytogenic treatment groups exhibited lower cecal lesion scores and reduced oocyst shedding than the infected control group. Among the phytogenic formulations evaluated, the combination containing 6 g/kg P. longum (T3) tended to produce the lowest numerical lesion score and oocyst shedding, although differences among phytogenic treatments were limited. Although some treatment groups also exhibited numerically favorable growth performance under challenge conditions, these observations should be interpreted cautiously because the study was designed primarily as a preliminary evaluation of anticoccidial efficacy, and feed intake and feed conversion ratio were assessed without independent pen-level replication. The observed biological responses may be associated, in part, with the improved dissolution characteristics afforded by the solid-dispersion formulation; however, this proposed mechanism was not directly investigated in the present study. In conclusion, solid-dispersion formulations containing C. longa and P. longum extracts demonstrated promising anticoccidial activity against experimental E. tenella infection and support further investigation as promising phytogenic candidates for complementary coccidiosis control strategies in broiler chickens.

Veterinary Sciences doi: 10.3390/vetsci13070674

Authors: Shushuai Yi Han Zhao Wanyi Li Yanmeng Liu Chao Yang Wanli Sha Jiangting Niu Baishuang Yin

Feline panleukopenia is an acute, highly contagious, and fatal infectious disease, posing a serious threat to the health and conservation of felids. It is necessary to develop diagnostic tools suitable for point-of-care testing (POCT). Herein, a colorimetric loop-mediated isothermal amplification (LAMP) assay targeting the FPV VP2 gene was developed using cresol red as a colorimetric indicator. The colorimetric LAMP enables result interpretation by visual observation of a color change from violet to yellow after amplification at 64 °C for 40 min. The limit of detection was 18.38 copies/µL for pMD-VP2 plasmid and 101.62 TCID50/mL for crudely extracted nucleic acids. This method exhibits no cross-reactivity with other common feline pathogens, and shows good repeatability. Nevertheless, this method is unable to differentiate FPV from canine parvovirus (CPV) owing to their high genetic similarity. Testing of 153 clinical samples revealed a 40.52% (62/153) positive rate, with 98.04% agreement compared to a commercial qPCR kit (42.48% positive rate). Collectively, the developed colorimetric LAMP assay offers a specific, sensitive, simple and visual approach for FPV detection, demonstrating great potential for field point-of-care applications.

Veterinary Sciences doi: 10.3390/vetsci13070673

Authors: Ning He Hongming Zhou Jiaming Yang Jiying Yin Qi Wang Zitong Jing Yang Liu Yuxin Kong Fanli Zeng Jianming Li Naichao Diao Kun Shi Rui Du

Bovine viral diarrhea virus (BVDV) infection is associated with inflammatory responses, but the mechanisms underlying inflammasome activation remain unclear. In this study, Madin–Darby bovine kidney (MDBK) cells were used to compare the inflammatory responses induced by the cytopathogenic NADL strain and the non-cytopathogenic TC strain. Both strains significantly increased IL-1β and IL-18 production and promoted NLRP3 inflammasome assembly, indicating activation of upstream inflammasome signaling. However, only the NADL strain markedly induced Gasdermin D (GSDMD) cleavage, membrane pore formation, lactate dehydrogenase release, and pyroptotic cell death. Further mechanistic analyses demonstrated that the NADL core protein C interacted with the inflammasome adaptor ASC, thereby facilitating NLRP3 inflammasome assembly and enhancing inflammatory cytokine secretion. Silencing ASC significantly impaired inflammasome activation and pyroptosis induced by protein C, confirming its essential role in this process. Collectively, these findings demonstrate that the cytopathogenic BVDV NADL strain activates ASC-dependent NLRP3 inflammasome signaling and GSDMD-mediated pyroptosis through its core protein C, providing a molecular explanation for the distinct inflammatory responses and pathogenic outcomes associated with different BVDV biotypes.

Veterinary Sciences doi: 10.3390/vetsci13070672

Authors: Lijun Shang Guanzhu Liu Shuang Wu Yan Ren Bo Liu Yunzhou Wang Tao Wang Lianyu Yang

Optimizing feeding regimes and finishing duration is essential for improving beef production efficiency and meat quality. This study evaluated the effects of two feeding regimes and three finishing durations on growth performance, nutrient digestibility, carcass traits, meat quality, blood biochemical parameters, rumen fermentation, and rumen microbiota in Black Angus bulls. A total of 552 healthy bulls with similar initial body weight were assigned to a 2 × 3 factorial design with two diets—a corn silage–rice straw–concentrate TMR (CSRC) and a rice straw–concentrate TMR (RSC)—and three finishing durations: 10, 12, and 14 months. The CSRC TMR increased average daily gain and reduced feed-to-gain ratio during early finishing (p < 0.05), whereas the RSC TMR resulted in greater final body weight and higher meat redness at 14 months (p < 0.05). Prolonged finishing decreased average daily gain, feed efficiency and nutrient digestibility (p < 0.05), but increased carcass weight, eye muscle area, backfat thickness and meat tenderness (p < 0.05). The CSRC TMR promoted higher total volatile fatty acid production (p < 0.05), an acetate-oriented fermentation pattern and greater rumen microbial alpha diversity. These findings support a stage-specific feeding regime for Black Angus bull finishing. Under the present conditions, corn silage inclusion may be used to improve early finishing efficiency, whereas rice straw–concentrate TMR may be more suitable for late finishing to balance final body weight, meat quality, and production cost.

Veterinary Sciences doi: 10.3390/vetsci13070671

Authors: Muhammad Zain Ghauri Ayesha Zafar M Khuzema Niaz Usman Nazir Asim Munir Muhammad Hamza Kiran Zahra Dejun Ji

Cashmere production declines with age in Inner Mongolian cashmere goats, but the molecular mechanisms are unknown. This study employed a transcriptome-wide RNA-seq analysis to compare age-associated and breed-related gene expression profiles in goat skin, with particular emphasis on age-dependent AKT1 expression and its associated signaling pathways. Skin tissues from cashmere goats at 12 and 15 months and age-matched non-cashmere controls were analyzed by RNA-seq (BGIDNBSEQ platform). Differential expression and functional enrichment analyses were performed on skin transcriptomes of cashmere and control goats at 12 and 15 months. Age-specific functional profiles emerged: 12-month cashmere goats showed enrichment in GTPase activity, glucose transport, and ribosomal assembly, whereas 15-month goats were enriched for autophagosome assembly and apoptosis. FoxO signaling was commonly enriched across both ages, while the Hepatitis B pathway was unique to 12 months. AKT1 was significantly upregulated at 12 months coinciding with peak cashmere production with reduced expression at 15 months. This age-dependent pattern was supported by coordinated regulation of downstream effectors, including TSC2, PIK3R2, MAPK9, and FOXO3. Collectively, these transcriptomic data provide a mechanistic framework linking age-dependent AKT1 regulation to the decline in cashmere fiber production.

Veterinary Sciences doi: 10.3390/vetsci13070670

Authors: Yueqi Wang Boyang Zhang Rui Yang Yan Zhang Daozhen Jiang Yifei Mao Bo Tang Xueming Zhang

Repeat breeding syndrome (RBS), characterized by repeated artificial insemination failure, is associated with ovarian dysfunction in ruminants and causes economic losses. Environmental endocrine-disrupting chemicals (EDCs), such as bisphenol F (BPF), may contribute to RBS, but the relationship between BPF exposure and RBS in cows remains unclear. In this study, we conducted an epidemiological investigation and clinical examinations in 111 adult Simmental cows, including 7 cows diagnosed with RBS, from two local farms, and measured the BPF levels in the drinking water, feed, and the serum samples. We further evaluated the protective effects of phillyrin, a bioactive compound derived from Forsythia suspensa, against BPF-induced ovarian toxicity using cow granulosa cells (CGCs). RBS mainly occurred in young cows with two or more parities. RBS cows showed signs of potential liver dysfunction, kidney injury, anemia, and fecal abnormalities suggestive of altered gut health, together with higher serum BPF levels and lower estradiol levels. Detectable BPF contamination in the drinking water and feed suggested environmental exposure as a possible contributor to RBS in cows. In vitro, BPF induced inflammatory and apoptotic responses in CGCs by upregulating pro-inflammatory and pro-apoptotic mediators, whereas phillyrin pretreatment markedly alleviated these effects. These findings suggest that BPF exposure might be associated with RBS in cows and phillyrin could be used as a promising natural agent for mitigating BPF-related reproductive toxicity.

Veterinary Sciences doi: 10.3390/vetsci13070669

Authors: Abd Ullah Liu Bing Dou Manna Pengshuai Li Kang Jiawei Yu Jie Muhammad Zahoor Khan Changfa Wang

Neonatal assessment in donkey foals is essential for reducing perinatal mortality and improving early management, yet breed-specific data remain scarce, particularly for indigenous Chinese breeds such as the Luxi Black donkey—an economically and genetically valuable breed of Shandong Province for which neonatal vitality has not been previously characterized. This prospective observational study aimed to describe the distribution of a modified four-parameter Apgar score and to summarize early physiological and morphometric parameters in Luxi Black donkey foals at birth. The study further explored whether these early parameters differed across Apgar score categories, with the aim of providing preliminary breed-specific reference data for neonatal assessment in this indigenous donkey breed. Fifty Jennies and the resulting 51 foals were enrolled at the National Black Donkey Breeding Center in Dong’e County, Shandong, China. Within 5 min of delivery, each foal was scored using a modified Apgar system (heart rate, respiratory rate, muscle tone, reflex response; maximum 8), and heart rate, respiratory rate, time to first suckling, rectal temperature, birth weight, body height, body length, chest girth, and abdomen girth were recorded. Foals were grouped according to their observed modified Apgar score categories (8/8, 7/8, 6/8, and 0/8). All 51 foals were included in the descriptive analysis of Apgar score distribution, while the three stillborn foals with an Apgar score of 0/8 were described separately and excluded from statistical comparisons. Physiological and morphometric parameters were compared only among live-born foals with Apgar scores of 8/8, 7/8, and 6/8. Of the 51 foals, 35 (68.63%) scored 8/8, 7 (13.73%) scored 7/8, 6 (11.76%) scored 6/8, and 3 (5.88%) were stillborn. Among live-born foals, respiratory rate (p = 0.008), time to first suckling (p < 0.001), and rectal temperature (p = 0.001) differed significantly among Apgar score categories, whereas heart rate showed no significant difference (p = 0.052). Birth weight and morphometric measurements did not differ significantly among live Apgar score categories. These preliminary findings suggest that the modified Apgar score may support early descriptive observation of neonatal status in Luxi Black donkey foals, although further studies with larger cohorts and objective outcome-based measures are needed.

Veterinary Sciences doi: 10.3390/vetsci13070668

Authors: Omar S. Saeed Sara A. Shabana Mahmoud Gamal Basem M. Ahmed Ayman H. El-Deeb Haitham M. Amer

Avian metapneumovirus (aMPV) is an important respiratory pathogen of poultry associated with respiratory disease, reproductive losses, secondary bacterial complications, and economic losses in commercial poultry systems worldwide. This narrative review summarizes current evidence regarding the epidemiology, molecular diversity, transmission ecology, diagnosis, vaccination, and wildlife–poultry interactions of aMPV across Africa and the Middle East. This literature was identified through structured searches of PubMed, Scopus, Web of Science, Google Scholar, and ScienceDirect using predefined keywords related to epidemiology, molecular characterization, surveillance, wild birds, diagnostics, and vaccination. Available evidence demonstrates widespread endemic circulation of aMPV, with subtype B predominating across most investigated poultry systems. However, major heterogeneity exists among studies regarding surveillance intensity, diagnostic methodologies, molecular characterization, and reporting standards. Current evidence suggests possible involvement of wild birds in viral dissemination, although definitive reservoir competence and transmission directionality remain insufficiently established. Important surveillance gaps persist across several African and Middle Eastern countries, emphasizing the need for harmonized surveillance, subtype-matched vaccination strategies, and integrated regional monitoring.

Veterinary Sciences doi: 10.3390/vetsci13070667

Authors: Breno Luis Nery Garcia Carlos Eduardo Fidelis Kristian da Silva Barbosa Gustavo Freu Marcos Veiga dos Santos

Antimicrobial resistance (AMR) is a global concern partly driven by excessive antimicrobial use (AMU) in animal production. Clinical mastitis (CM) is a leading cause of AMU in dairy farms, highlighting its importance from a public-health perspective. This observational study aimed to evaluate the use of antimicrobial susceptibility testing as a criterion for selecting antimicrobials for the treatment of mild and moderate CM in dairy cows. To this end, the association between in vitro antimicrobial susceptibility test results of mastitis-causing pathogens and in vivo bacteriological cure (BC) was assessed. A total of 340 cows with mild or moderate CM from 14 commercial dairy herds in Brazil were enrolled in the study. Isolates from the pretreatment milk samples were subjected to antimicrobial susceptibility testing (AST) using agar disk diffusion. Susceptibility results and cow-level variables were included in a generalized linear mixed model (GLMM) to evaluate their association with BC. No significant association was found between the antimicrobial susceptibility results and BC of the mastitis cases. In conclusion, although in vitro susceptibility testing remains an important tool for AMR surveillance, the findings of this study suggest that its utility in guiding antimicrobial treatment decisions in CM cases is limited.

Veterinary Sciences doi: 10.3390/vetsci13070666

Authors: Annika Gersemann Gerald Reiner Sonja von Berg

Extraintestinal pathogenic Escherichia coli (ExPEC) are opportunistic pathogens in pigs and can cause systemic infections with neurological signs. Cases in suckling piglets have rarely been reported. In November 2024, suckling piglets aged 1–3 weeks on a commercial pig farm developed neurological signs, including altered consciousness, vertical nystagmus, ataxic gait, and recumbency. Within two affected batches comprising 1150 and 1200 piglets, 17 and 26 animals, respectively, developed clinical signs (morbidity 1.48% and 2.17%). Initial treatment with amoxicillin and dexamethasone was ineffective. Early administration of intramuscular enrofloxacin combined with dexamethasone was associated with clinical improvement in approximately 50% of affected piglets. Pathological examination revealed atelectatic lung regions, dilated small intestines with yellowish content, and moderate mesenteric vessel congestion. Histologically, the central nervous system of both piglets exhibited hyperemia, edema, disseminated diapedesis hemorrhages, thrombi, and intravascular rod-shaped bacteria. E. coli was isolated from the CNS, lungs, pericardium, and joints. Molecular analysis confirmed the presence of virulence-associated genes fimA, fimH, and iucD. Differential diagnostic PCRs for Streptococcus suis, Glaesserella parasuis, and Mycoplasma hyorhinis were negative. This case report documents the first occurrence of an ExPEC-like E. coli infection in this herd. The documented clinical, pathological, and molecular findings support the importance of considering ExPEC-like E. coli as a differential diagnosis in therapy-resistant neurological diseases in piglets.

Veterinary Sciences doi: 10.3390/vetsci13070665

Authors: Ciprian Ober Vasile Coza Ballai Szidónia Bogdan Tancău Christos Yiapanis William McCartney Denis Gaceu Ștefana Mureșan Maria Moraru Bianca Boldiș

Grade IV lateral patellar luxation is an advanced orthopedic condition that may be associated with severe skeletal deformities, chronic soft-tissue contracture, and irreversible loss of normal stifle function. Although reconstructive procedures are considered the treatment of choice in most dogs, some end-stage cases may not be amenable to conventional correction. This report describes the clinical presentation, surgical management, and long-term outcome of a young dog with severe bilateral grade IV lateral patellar luxation treated with single-stage bilateral stifle arthrodesis. A 1-year-old, 42-kg intact female Romanian Shepherd Dog was presented with chronic severe hindlimb dysfunction, crouched posture, and permanent bilateral patellar luxation. Orthopedic and radiographic examination revealed severe femoral valgus, proximal tibial varus, external tibial torsion, absence of a functional trochlear groove, marked caudal thigh muscle contracture, and severely restricted stifle extension. Computed tomography was considered but not performed because of financial limitations. Based on the severity of the deformities and the limited preoperative range of motion, conventional reconstructive procedures were considered unlikely to restore functional joint motion. Bilateral stifle arthrodesis was therefore performed during a single anesthetic event using tibial tuberosity osteotomy, temporary Kirschner-wire fixation, and bilateral 3.5-mm locking compression plates. Due to severe chronic soft-tissue contracture, final arthrodesis angles of 95° and 99° were achieved. Progressive clinical improvement and radiographic bone union were observed throughout follow-up. At 12 months, complete bilateral arthrodesis and implant stability were confirmed radiographically. At 24 months, the dog remained independently ambulatory with a markedly improved quality of life despite persistent mechanical gait abnormalities, including circumduction and intermittent bunny-hopping. This case suggests that bilateral stifle arthrodesis may represent a salvage option in carefully selected dogs with end-stage bilateral grade IV lateral patellar luxation when conventional reconstructive procedures are unlikely to restore useful limb function.

Veterinary Sciences doi: 10.3390/vetsci13070664

Authors: Mercédesz Adrienn Veres Zsófia Anna Tóth Enikő Illés Patrik Mag Eszter Kaszab Enikő Fehér Ákos Jerzsele Ádám Kerek

Background: Canine otitis externa is a clinically relevant niche where Pseudomonas aeruginosa may persist under inflammatory, antimicrobial, and biofilm-favoring conditions. This study characterized the virulome and population structure of canine otitis externa-derived P. aeruginosa isolates from Hungary. Methods: Of 110 isolates previously subjected to minimum inhibitory concentration testing, 70 phenotypically selected isolates underwent long-read whole-genome sequencing and were included in virulence-factor analysis. Virulence-associated genes were detected using the Virulence Factor Database with an 80% identity and 80% coverage threshold and were interpreted by functional systems, including motility, adhesion, biofilm formation, secretion systems, siderophores, quorum sensing, secreted enzymes, and type III secretion system effectors. Results: The isolates carried a broad and highly conserved virulence-associated backbone. Genes related to alginate production, type IV pili, flagellar motility, pyochelin, phenazine biosynthesis, Xcp secretion, type VI secretion, and proteolytic enzymes were widely distributed. Type III secretion effector genes were variable: exoT was detected in 68/70 isolates, exoY in 63/70, exoS in 50/70, and exoU in 15/70. MLST indicated a genetically diverse population, and virulome size showed no strong correlation with the modified multiple antimicrobial resistance index. Conclusions: These findings support integrated genomic surveillance of virulence and antimicrobial resistance in canine otitis-associated P. aeruginosa.

Veterinary Sciences doi: 10.3390/vetsci13070663

Authors: Gang Zhou Ying Liu Xuanxuan Pu Qiugui Ning Xiaoshan Guo Liwei Wang Yuhong Zhong Guolian Wang Xuefeng Guo Mengzhi Wang

Alterations in microbiota, transcript and metabolites are critical to intestinal homeostasis and host health. This study used a combination of 16s rRNA, transcriptome sequencing and liquid chromatography–mass spectrometry to investigate intestinal microbiota, genes and metabolic profiles in the ileum of Hu sheep fed broccoli extract. Here, we randomly allocated 14 Hu sheep to two diets: a basal diet without any supplementation (NC) and a basal diet supplemented with 200 mg/kg broccoli tail (BT). After 60 days of treatment, blood and jejunal samples were collected for serum biochemical indicators and multi-omics analysis. In this study, the extract of broccoli tails had a significant effect on the serum biochemical indicators, including white blood cells, red blood cells, mean corpuscular volume, mean corpuscular hemoglobin concentration, mean platelet volume, triglycerides and total protein in Hu sheep (p < 0.05). Transcriptomic analysis showed that the 672 differentially expressed genes between the NC and BT groups were primarily enriched in linoleic acid metabolism, steroid hormone biosynthesis, and cholesterol metabolism. Metabolomics analysis using Kyoto Encyclopedia of Genes and Genomes enrichment showed that the 41 differentially abundant metabolites were mainly enriched in bile secretion, vitamin B6 metabolism, and the mTOR signaling pathway. 16S rRNA sequencing results indicated that the extract of broccoli tails increased the relative abundance of Peptostreptococcaceae and decreased the relative abundance of Lachnospiraceae, Lachnospirales, and Bacteroidaceae. Integrated transcriptome, metabolome, and microbiome analysis showed that the gut microbiota and host transcriptomic changes may participate in systemic metabolic regulation by modulating amino acid metabolism, lipid signal transduction, nucleotide metabolism, and vitamin B6-related metabolic pathways. These findings demonstrate that the extract of broccoli tails modulates intestinal gene expression, systemic metabolism, and gut microbial ecology in Hu sheep, providing new insights into the utilization of agricultural byproducts as a functional feed supplement for ruminants.

Veterinary Sciences doi: 10.3390/vetsci13070662

Authors: Mingyang Jin Jingkang Huang Chao Qin Kaixuan Yang Fuquan Xiao He Meng

Primordial germ cells (PGCs) are the earliest precursors of gametes and essential cellular materials for poultry germplasm conservation and genetic modification. In this study, PGCs isolated from 2.5-day male and female Silkie chicken embryos were defined as circulating PGCs (cPGC_M and cPGC_F), whereas PGCs isolated from 8.5-day male and female embryos were defined as gonad-derived PGCs (gPGC_M and gPGC_F). RNA sequencing with three biological replicates per group was performed to characterize developmental-stage- and sex-associated transcriptional programs. Comparative transcriptomic analyses identified stage-associated differences in pathways related to cell–matrix interaction, focal adhesion, PI3K–Akt signaling, and stem cell pluripotency, with more differentially expressed genes detected in the female than in the male stage comparison. Sex-biased expression was detectable at the circulating stage, mainly reflecting W- and Z-linked expression differences, and the number of sex-biased genes was greater after gonadal colonization, primarily because more autosomal differentially expressed genes were detected. Candidate genes and pathways, including HINTW, DMRT1, SOX2, EDNRB, LPAR4, GFRA3, ECM–receptor interaction, ribosome-related modules, and calcium signaling, were associated with these comparisons and are presented as targets for future validation rather than as confirmed regulators. Because the study used three biological replicates per group, RT-qPCR corroboration was limited to six female stage-associated genes, and no functional perturbation assays were performed, the findings are descriptive and do not establish causal mechanisms of PGC migration, gonadal colonization, or sex differentiation. Within these limitations, the dataset provides an exploratory resource for subsequent studies of chicken PGC biology and biotechnology.

Veterinary Sciences doi: 10.3390/vetsci13070661

Authors: Yanhan Lin Xiaoyang Zhu Yifan Meng Jiachun Zou Wanying Xie Shuai Yang Meng Cui Ming Qiu Xinkai Wang Qinchao Guan Hong Lin Sen Jiang Wanglong Zheng Jianzhong Zhu Kewei Fan Nanhua Chen

Feline and canine coronaviruses (FCoVs and CCoVs) are widely prevalent in companion animals. The recent emergence of a highly virulent FCoV-CCoV recombinant virus, FCoV-23, caused a rapid feline infectious peritonitis (FIP) outbreak in Cyprus, which raises a serious concern about the cross-species transmission of CoVs among companion animals. Here, we evaluated the prevalence, transmission and evolution of FCoV and CCoV in the Jiangsu and Zhejiang provinces of China in 2025. A universal RT-qPCR assay for FCoV and CCoV was established and used for the detection of 700 clinical samples, including 501 from cats and 199 from dogs. A total of 92 samples (13.14%, 92/700) were detected as positive, containing 79 positive samples (15.76%, 79/501) from cats and 13 positive samples (6.5%, 13/199) from dogs. Ten complete S genes and two complete genomes were obtained from positive samples. Fragment comparison and phylogenetic analysis identified one CCoV-I sequence from a feline sample and one FCoV-II sequence from a canine sample. Furthermore, recombination analyses not only detected intra-species recombination events but also inter-species cross-over events. Collectively, this study, for the first time, revealed that cross-species transmission and recombination events occurred between FCoV and CCoV among companion animals in the Jiangsu–Zhejiang region of China in 2025.

Veterinary Sciences doi: 10.3390/vetsci13070659

Authors: Aaron C. Ericsson

Molecular microbiology has revolutionized our understanding of the complex host-associated microbiomes required for normative development and physiology. Horses and other members of the family Equidae are particularly reliant on the early maturation and lifelong maintenance of an unusually rich hindgut microbiome for optimal digestion and overall health and performance. Research on the equine gut microbiome has accelerated in the past several years, necessitating a renewed appraisal of the field. The present work is a comprehensive and critical review of the literature regarding the bacterial gastrointestinal microbiome of horses. First, the developmental trajectory of the foal gut microbiome is discussed, followed by descriptions of the taxonomic membership of the core equine gut microbiome, its primary functions and effects on host physiology, and intrinsic and extrinsic factors that shape the equine microbiome during health, with a focus on diet and supplements. Next, evidence supporting adverse effects on the equine gut microbiome of gastrointestinal conditions including colic and colitis, extraintestinal conditions including obesity and laminitis, and pharmacological interventions including antibiotics and non-steroidal anti-inflammatory drugs is summarized. Lastly, clinical and experimental research investigating the effects of treatments targeting the gut microbiome of horses, including probiotics, prebiotics, and fecal microbiome transfer, is critically examined. Conclusions summarize the connection between natural (i.e., wild) equine behavior and the health of the equine gut microbiome and the impacts of human management.

Veterinary Sciences doi: 10.3390/vetsci13070660

Authors: Rhoda C. Apitzsch Sarah Handel Konstanze Krueger

There has been little research on the social behaviour of stallions and geldings kept together in group housing systems. The aim of this study was to investigate differences in behaviour and movement activity between stallions and geldings kept together in group housing. Behavioural observations and GPS measurements were conducted in Germany from July to September 2024 in four mixed groups of horses in Germany. The groups comprised 35 animals, 18 stallions and 17 geldings. The horses were observed for three consecutive days over eight hours per day. No injuries or stereotypic behaviours were observed throughout the observation period. Stallions displayed significantly higher levels of affiliative, agonistic, dominance, ritualised, reproductive and play behaviours than geldings (all p < 0.05). However, attack, threat, comfort, avoidance, and resting behaviours, as well as movement activity did not differ significantly between stallions and geldings. Greater space availability per horse was associated with increased comfort, resting, reproductive and play behaviours, and dominance behaviour decreased and attacks tended to decrease with increasing space availability. The horses moved an average distance of 2.36 km during the observation periods.

Veterinary Sciences doi: 10.3390/vetsci13070658

Authors: Meihe Zhao Tingyu Li Liyin Du Qinghua Deng Jingdong Mao Zhenwei Jia Yuming Zhang

Bovine Mycoplasma bovis pneumonia (MBP) is an important component of bovine respiratory disease, and its management is complicated by persistent infection and antimicrobial stewardship concerns. Houttuynia cordata Thunb. has reported anti-inflammatory and immunomodulatory activities, but its potential host-directed mechanisms in MBP remain unclear. This in silico study used network pharmacology and molecular docking to identify candidate compounds, common drug–disease targets, enriched biological functions, and predicted ligand–target interactions. A total of 145 putative targets of H. cordata and 474 MBP-associated disease targets were obtained from TCMSP, GeneCards, OMIM, and CTD, yielding 43 common drug–disease targets. Dual-confidence STRING analysis, cytoHubba ranking, and MCODE module analysis prioritized TNF, IL6, IL1B, PTGS2, PPARG, IFNG, CASP3, and MMP9 as candidate core targets. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment indicated convergence on cytokine-mediated signaling, inflammatory response, immune regulation, oxidative stress, IL-17 signaling, and TNF signaling. Molecular docking suggested favorable predicted interactions for quercitrin–PTGS2, quercetin–TNF, quercetin–IL6, and quercitrin–CASP3. These computational findings suggest that H. cordata may be associated with host inflammatory and immune-response modulation in MBP, mainly through flavonoid-related interactions with inflammation- and apoptosis-related targets. Further bovine-specific experimental validation is required before biological activity or practical application can be inferred.

Veterinary Sciences doi: 10.3390/vetsci13070657

Authors: Mingyue Wan Weizhen Shen Peng Wang Mengran Zhang Jing Chen Bin Zhou

Although commercial vaccines against porcine circovirus type 2 (PCV2) have been widely implemented globally, PCV2 remains endemic in swine populations, accompanied by ongoing genotype replacement. Meanwhile, the emergence of novel porcine circoviruses (PCVs), including PCV3 and PCV4, has further complicated the prevention and control of porcine circovirus-associated diseases (PCVAD). This study systematically characterized the epidemiological patterns and genetic diversity of PCVs circulating in Eastern China. A total of 739 clinical samples collected between 2010 and 2016 were screened for PCV2. Additionally, 653 samples obtained during 2023–2024 were analyzed using a triplex real-time quantitative PCR (qPCR) assay for the simultaneous detection of PCV2, PCV3, and PCV4. Full-genome amplification and sequencing were subsequently performed on all PCR-positive samples. Epidemiological analysis revealed an overall PCV2 positivity rate of 37.62% during 2010–2016. In the 2023–2024 cohort, the positivity rates for PCV2 and PCV3 were 35.99% and 16.39%, respectively, with a co-infection rate of 10.26%. Notably, no PCV4-positive samples were detected. Phylogenetic analysis demonstrated that PCV2d is the predominant genotype in Eastern China. Furthermore, PCV2g strains were identified in clinical samples for the first time in mainland China, while PCV3b were determined to be the dominant circulating subtype of PCV3. Multiple critical amino acid substitutions were identified within the neutralizing epitopes of the PCV2 Cap protein, and a recombination event involving a PCV2d vaccine strain and a PCV2c reference strain was detected. In contrast, the PCV3 genome exhibited a high degree of genetic conservation. Collectively, these findings expand the molecular epidemiological landscape of PCVs in Eastern China and elucidate the evolutionary dynamics of circulating PCV strains, providing important insights for the development of next-generation vaccines and region-specific PCVAD control strategies.

Veterinary Sciences doi: 10.3390/vetsci13070656

Authors: Shuang Chen Yikuan Qian Lixiang Wei Ming Wu Yu Yang Xuepeng Cai Jie Li Qingling Meng Jun Qiao

This study aimed to identify and characterize a high-efficiency chitinase gene from Brevibacillus laterosporus (B. laterosporus), characterize its enzymatic traits, and assess its degrading activity against Caenorhabditis elegans (C. elegans) and Parascaris equorum (P. equorum) eggs. A nematicidal B. laterosporus isolate was subjected to whole-genome sequencing for chitinase gene screening, cloning, and molecular identification. The target gene was heterologously expressed in Escherichia coli BL21 (DE3), and biochemical properties including temperature, pH, metal ions, and substrate conditions were investigated, its kinetic parameters were determined, and its biological effects on C. elegans and P. equorum eggs were analyzed. The protein BLChi79 (79.3 kDa) belongs to GH18 chitinases with a typical carbohydrate-binding module. Its optimum activity occurred at 60 °C and pH 6.0, with colloidal chitin as the optimal substrate. Mg2+, Fe2+, and Mn2+ boosted its activity, while K+, Cu2+, Zn2+, and Ca2+ suppressed catalysis. Its Km was 6.14 mg·mL−1, Vmax 7.78 μmol·min−1·mg−1, and kcat 10.16 min−1. The purified recombinant enzyme could degradethe C. elegans chitin layer and the P. equorum egg vitelline envelope. In summary, the BLChi79 derived from B. laterosporus targets chitin-enriched eggshells, acting as a green biocatalyst for controlling livestock gastrointestinal nematodes.

Veterinary Sciences doi: 10.3390/vetsci13070655

Authors: Shiwen Gao Guoqing Li Xiangyu Wang Weifang Gu Dingnuoya Guo Zongping Xian Xuelian Ma Jun Meng Hongqiong Zhao Lu Liu

Rhodococcus equi (R. equi) primarily induces fatal pulmonary and extrapulmonary pyogenic granulomatous infections in foals, imposing substantial economic burdens on the equine industry. The emergence and spread of multidrug-resistant (MDR) R. equi have led to a therapeutic impasse in clinical settings. Although vaccination is a proven strategy against MDR pathogens, no commercial vaccine is currently available for R. equi. In this study, we employed a bioinformatics approach to systematically identify and prioritize antigenic epitopes derived from R. equi for multi-epitope vaccine design. Using ABCPred, NetMHCpan EL, and IEDB servers, 27 MHC-I and 9 MHC-II epitopes were selected from five previously validated R. equi vaccine candidates: ABC transporter, PBD2, NlpC/P60, Esterase, and M23. These epitopes were coupled with distinct peptide linkers to construct six multi-epitope vaccine constructs, designated V1–V6. The physicochemical properties, antigenicity, immunogenicity, and toxicity of the six vaccine constructs were analyzed, and the V3 and V4 constructs were ultimately selected. Using the HDOCK and Gromacs tools, the intermolecular interactions, binding affinity, and thermal stability of the V3 and V4 constructs with the equine MHC molecules EQCA-I and EQCA-II were evaluated. The results confirm that V3 and V4 exhibit strong binding affinity to EQCA-I and EQCA-II, with stable conformations following binding, indicating theoretical potential to induce humoral and cellular immunity in foals. Recombinant plasmids for V3 and V4 were constructed, and the V3 and V4 proteins were successfully prepared, confirming the feasibility of prokaryotic expression for these vaccine constructs. Immunization assays in SPF BALB/c mice showed that the multi-epitope vaccines elicited robust antigen-specific IgG antibody responses, reflecting preliminary humoral immunogenicity. However, these murine data have translational limitations, as they cannot fully represent equine immune responses. The findings establish a crucial theoretical foundation for the advancement of vaccines targeting R. equi while offering a reference for the design of vaccines against other drug-resistant microbial pathogens.

Veterinary Sciences doi: 10.3390/vetsci13070654

Authors: Shiyu Duan Zixuan Wang Yuchen Jia Xin’er Lan Cong Peng Xiyue Deng Hui Jiang Wei Wang Guangzhi Zhong Yiping Zhu Jing Li

Equine osteoarthritis (OA) is a major cause of lameness and economic loss in horses. While platelet-rich plasma (PRP) has clinical potential, the biological effects of fresh PRP (F-PRP) and freeze-dried PRP (FD-PRP) remain insufficiently defined. This pilot study compared 25% and 50% F-PRP and FD-PRP in an interleukin-1β-induced equine cartilage-synovium explant co-culture model. PRP treatments reduced inflammatory responses, with significant downregulation of COX-2 and PGE2 expression, and 25% F-PRP showed the most consistent inhibition of nitric oxide production. PRP also significantly reduced glycosaminoglycan release and altered matrix-related gene expression; however, FD-PRP significantly upregulated MMP13, indicating a potential pro-catabolic response. Untargeted LC/MS metabolomics showed that F-PRP and FD-PRP were associated with changes in glucose, purine, amino acid, lipid, and nucleotide metabolism. Growth factor analysis further showed lower PDGF and TGF-β1 concentrations in FD-PRP than in F-PRP. Overall, F-PRP showed more consistent anti-inflammatory and matrix-protective effects, whereas FD-PRP requires further optimization and safety validation before clinical application.

Veterinary Sciences doi: 10.3390/vetsci13070653

Authors: Nadia Stoppani Federica Raspa Edoardo Fiorilla Sandra Maione Achille Schiavone Cecilia Mugnai Dominga Soglia

In this study, an exploratory transcriptomic investigation was conducted to evaluate the feasibility of using feather transcriptomics to detect sex differences and gene responses to physiological changes in chickens. Feathers represent a promising non-invasive biological source of RNA, as the feather pulp of growing feathers contains living cells capable of active transcription. Growing feathers were collected from 150-day-old male and female chickens (Bionda Piemontese, a slow-growing breed) raised under a free-range system and fed two finisher diets differing in lipid content: low-lipid (LL, ether extract 3.6%) and high-lipid (HL, ether extract 9.3%) diets. RNA was extracted from feather pulp, and 12 pools were subjected to whole RNA-Seq analysis. The study was designed as 2 × 2 factorial experiments investigating the effects of diet and sex on gene expression. A total of 17,360 transcripts were detected and used for downstream analyses. Differential gene expression and functional enrichment analyses were performed. The main effects of diet and sex were estimated with an additive design using the DEseq2 package, while for the sex-specific diet analyses, subgroup comparisons were conducted on the RaNA-Seq platform. The analysis of the main effect of diet reveals that three genes associated with ether lipid metabolism (PLA2G10, PLA2G4F, and ENPP6) were upregulated in chickens fed the HL diet. In roosters, HL feeding significantly altered the expression of APOA1 and SLC27A4, suggesting an effect on lipid transport and metabolic regulation within the PPAR signaling pathway. In contrast, hens showed differential expression primarily in pathways related to apelin signaling, extracellular matrix remodeling, and cardiovascular function, rather than classical lipid metabolism pathways; additionally, gene set enrichment analysis indicated a limited enrichment of linoleic acid metabolism, suggesting secondary involvement of lipid metabolic processes. These findings are consistent with those in the literature reporting sex-related differences between males and females. The results further suggest that transcriptomic responses to dietary lipid supplementation can be investigated through the expression of selected candidate genes in feather pulp. Among the genes identified, PLA2G10, PLA2G4F, ENPP6, APOA1, and SLC27A4 emerged as potential molecular markers associated with dietary treatment, and the importance of sex-dependent transcriptional responses was highlighted. In conclusion, this study demonstrates the potential of feather pulp as a viable source of RNA for transcriptomic analyses in live chickens, providing a minimally invasive alternative to conventional tissue sampling. These preliminary results also support the hypothesis that feathers represent a practical and ethically favorable tissue for future nutrigenomic and genetic improvement studies, ultimately supporting more sustainable poultry production.

Veterinary Sciences doi: 10.3390/vetsci13070652

Authors: Jianzhong Wang Hang Yan Jing Liu Rui Zhou Wei Yin Jia Zhong Panpan Sun Na Sun Zhenbiao Zhang Yaogui Sun Huizhen Yang Kuohai Fan Hongquan Li

The search for effective alternatives or adjuncts to conventional anti-infective strategies in food animals has increased interest in plant-derived bioactive compounds. Matrine, a quinolizidine alkaloid derived from Sophora flavescens, has attracted considerable attention because of its broad bioactivities and potential veterinary relevance. Recent oral pharmacokinetic and intestinal-lumen PBPK studies in pigs have begun to emerge; however, currently available swine data remain largely confined to oral exposure-oriented experiments and do not permit direct route comparison or estimation of absolute oral bioavailability. In the present study, the plasma pharmacokinetics of matrine in pigs were evaluated after single-dose intravenous and oral administration. Twelve healthy piglets received matrine at 8 mg/kg body weight via either the intravenous or oral route, and plasma concentrations were quantified using a validated UPLC–MS/MS method. Pharmacokinetic parameters were estimated by non-compartmental analysis. Compared with intravenous administration, oral dosing produced lower observed exposure based on AUC0−t and a lower peak plasma concentration, with AUC0−t and Cmax values of 418.94 ± 75.52 h·ng/mL and 66.24 ± 8.44 ng/mL, respectively, versus 558.01 ± 59.57 h·ng/mL and 224.64 ± 20.94 ng/mL after intravenous administration. Oral administration was associated with a Tmax of 2.49 ± 0.02 h and a longer apparent terminal half-life and mean residence time than intravenous dosing. These findings provide pharmacokinetic evidence to support future route selection, dosing-regimen design, and PK/PD-based dose optimization of matrine in pigs.

Veterinary Sciences doi: 10.3390/vetsci13070651

Authors: Lingling Chen Wanying Gao Yang Song Zihao Huang Jialing Long Jiaqi Nie Zengliang Wang Shulei Jia

Background: This study aims to delineate the present and projected suitable habitats for four dominant tick species in the Beijing–Tianjin–Hebei (BTH) region, providing a spatial basis for targeted tick-borne disease surveillance. Methods: We systematically reviewed the published literature and the Global Biodiversity Information Facility (GBIF) to compile tick occurrence records in the BTH region. A total of 167 geo-referenced occurrence records with verified coordinates were obtained for four dominant species: Haemaphysalis longicornis, Haemaphysalis concinna, Dermacentor silvarum, and Ixodes persulcatus. The MaxEnt model was applied with bioclimatic variables (WorldClim, 2.5 arc-min), elevation, slope, aspect, and NDVI. Model performance was evaluated using the area under the receiver operating characteristic curve (AUC) for within-species comparisons, complemented by the True Skill Statistic (TSS), Cohen’s kappa, and omission rate. Future projections (2021–2040, 2041–2060, 2061–2080, 2081–2100) were made under the SSP245 scenario using only climate variables, as the NDVI and topographic variables cannot be reliably forecast. Results: The four dominant tick species showed distinct distribution patterns: Hae. longicornis was widely distributed across the BTH region, whereas Hae. concinna, D. silvarum, and I. persulcatus were mainly found in the northern and northwestern mountainous areas. The primary environmental drivers were temperature, elevation, and the NDVI. MaxEnt models achieved good predictive performance (test AUC: 0.86–0.91; TSS: 0.72–0.88). Under future climate scenarios, suitable habitat centroids were projected to shift northwestward for Hae. longicornis (~57.6 km), D. silvarum (~71.1 km), and I. persulcatus (~50.0 km), and northeastward for Hae. concinna (~63.0 km) by 2081–2100. Conclusions: In this study, we identified current and future high-risk areas for four dominant tick species in the BTH region, providing a reproducible foundation for surveillance. Future projections should be interpreted with caution as they only account for climatic changes.

Veterinary Sciences doi: 10.3390/vetsci13070650

Authors: Lin Cheng Di Wang Jia-Rui Zhang Yi-Han Zhang Xin-Rui Wu Ya-Mei Huang Min Li Fu-Yan Wang Yang Zhao Xin-Feng Han Min Cui Yong Huang Jing Xia

Outbreaks of avian infectious bronchitis virus (IBV) often occur in vaccinated flocks. The antibody-dependent enhancement (ADE) has been proposed as a potential mechanism underlying coronavirus vaccine failure. However, this hypothesis has yet to be substantiated in flocks. This study demonstrates ADE occurrence in IBV (gamacoronavirus) in vitro and in vivo. Using the SPF chicken host model, primary infection with an O-glycosylation-modified attenuated strain enhanced pathogenesis upon secondary homologous/heterologous virulent challenge, increasing morbidity/mortality (≥30%), pathological lesions, and viral loads. Notably, sequentially attenuated infections also induced ADE, suggesting live attenuated vaccine risks. The immune serum raised against the O-glycosylation-modified attenuated strain was also pre-mixed with the challenge strain, and the mixtures were then inoculated into target cells, non-susceptible macrophage cells, or a co-culture of both cell types. The serum-virus complexes replicated poorly in macrophages, yet immune cells amplified the expression of inflammatory factors and ADE-mediated viral replication in target cells, indicating a significant promoting role of immune cells in this process. The concentrations of complement component C3 and neutralizing antibodies in the immune serum were also measured, and results showed that the induction of this ADE is associated with high complement component C3 and low neutralizing antibody titers. These findings highlight risks for vaccines and antibody-based therapeutic strategies of coronavirus infection.

Veterinary Sciences doi: 10.3390/vetsci13070649

Authors: Ling Zhang Zichang Liu Junjun Du Xuesong Zhang Fangzhe Ren Yuanyue Tang Xinan Jiao Jinlin Huang

Campylobacter jejuni (C. jejuni) is a prevalent zoonotic pathogen that poses a significant threat to global public health, causing gastroenteritis with symptoms ranging from mild diarrhea to severe complications, and is widely distributed among various animal hosts, including poultry, livestock, and wild birds. Among these, wild birds, characterized by their exceptional species diversity, high mobility, and long-distance migratory behavior, are capable of disseminating infectious agents across geographic regions and ecological settings. However, wild birds are increasingly recognized as potential carriers, research on C. jejuni in these hosts remains nascent and fragmented, and a unified understanding has yet to emerge. Therefore, this review synthesizes current knowledge on the prevalence patterns, key determinants, and genetic and transmission characteristics of wild bird origin C. jejuni, aiming to provide a foundation for subsequent in-depth evaluation of the public health significance of C. jejuni derived from wild birds.

Veterinary Sciences doi: 10.3390/vetsci13070648

Authors: Kyuman Cho

A comparative study on hemicircular tibial plateau leveling osteotomy (hTPLO) and TPLO for correcting cranial cruciate ligament (CCL) ruptures at excessive tibial plateau angles (eTPAs) was performed with a specially fabricated hTPLO saw blade (hemicircular) and TPLO saw blade (quartercircular), respectively. Ten 3D-printed bone models with eTPAs (TPA > 40°), were resized to accommodate the respective saw blade shape for each method. Cranial closing wedge osteotomy (CCWO) combined with TPLO was also performed. Distances of segment rotation below the patellar tendon insertion (PTI) and differences between anatomical and mechanical axes after correction were measured. The TPAs of the bone models were 46.06 ± 4.40° (40.0–50.7°). Postoperative TPAs of hTPLO, TPLO, and TPLO combined with CCWO were 4.93 ± 1.025°, 4.52 ± 0.085°, and 4.17 ± 1.128°, respectively. Rotation distances from the PTI were 0.46 ± 2.153 mm, 8.67 ± 2.318 mm, and 1.13 ± 1.796 mm, respectively. hTPLO showed significantly less movement of the segment below the PTI than TPLO with or without CCWO (p < 0.01). Differences between anatomical and mechanical axes after hTPLO and TPLO with or without CCWO were 5.74 ± 2.231°, 6.43 ± 2.105°, and 5.16 ± 2.465°, respectively. Within the limitations of this model-based study, hTPLO demonstrated geometric feasibility for correction of excessive tibial plateau angles and was associated with less rotational displacement of the proximal tibial segment relative to the patellar tendon insertion than conventional TPLO and TPLO combined with CCWO. Further cadaveric, biomechanical, and clinical studies are required to evaluate its safety and clinical applicability.

Veterinary Sciences doi: 10.3390/vetsci13070647

Authors: Qiao-Qiao Zhang Hao-Jie Zhang Lan-Lan Zheng Yue Zhang Hong-Ying Chen Shi-Jie Ma

Porcine epidemic diarrhea virus (PEDV) remains one of the most critical enteric coronaviruses affecting the global swine industry. PEDV causes severe diarrhea, dehydration, and high mortality in neonatal piglets. Despite the widespread use of commercial vaccines, persistent PEDV outbreaks worldwide indicate that current vaccination strategies provide suboptimal protection. Increasing evidence suggests that the limited effectiveness of current control strategies is primarily attributable to two interrelated factors: continuous antigenic variation among circulating PEDV strains and inadequate induction of maternal mucosal immunity. Unlike systemic viral infections, effective protection against PEDV in neonatal piglets predominantly depends on lactational immunity mediated by the gut–mammary gland–secretory IgA (sIgA) axis. However, most currently available vaccines predominantly induce systemic IgG responses and fail to effectively stimulate intestinal immune imprinting or sustain sIgA secretion in colostrum and milk. In this review, we summarize the mechanisms underlying PEDV evolution and maternal mucosal immunity, with particular emphasis on the gut–mammary gland–sIgA axis. We further discuss the recent advances in and limitations of current vaccine platforms and propose an integrated framework for broadly protective PEDV vaccines based on structural antigen optimization, mucosal-targeted immunization, and sIgA-oriented evaluation systems. This framework may provide new insights into the rational design of more effective maternal vaccines against PEDV and other enteric coronaviruses affecting pigs.

Veterinary Sciences doi: 10.3390/vetsci13070646

Authors: Ericka L. Carroll Nouran Nawar Melissa Bourgeois

This online survey of 1330 veterinarians and 5071 cat owners across 14 countries aimed to identify drivers, barriers and unmet needs regarding feline vaccination compliance. Veterinarians and cat owners differed on the perceived value of vaccination; 16% of cat owners cited vaccination as a key contributor to feline health compared with 69% of veterinarians. Cat owners reported that 71% of their cats had seen a veterinarian within the previous year and 65% were previously vaccinated, aligning with veterinarian-reported compliance rates of 77% for core vaccines. Primary vaccination drivers for cat owners were the desire to maintain their cat’s health (62%) and a veterinarian’s recommendation (57%); veterinarians perceived vaccine reminders (71%) as the top driver. Owners who elected not to vaccinate their cats believed vaccines were unnecessary for healthy (27%) or indoor cats (27%). This aligned with veterinarian-reported barriers: lack of client understanding (53%), vaccine misinformation (51%) and antivaccine attitudes (51%). Most cat owners (58%) cited their veterinarian as their primary vaccine information source. However, only 23% of veterinarians report discussing the importance of vaccination with cat owners. Cat owners prioritized low side effects (88%), multi-antigen protection (86%) and >1 year of protection (86%; 74% want 3 years). Concerning price and compliance, 87% (61% yes, 26% maybe) would pay more for long-lasting protection, and 91% (66% yes, 25% maybe) of vaccinating owners would attend annual wellness visits even when vaccination is not due. By addressing educational gaps, communicating the role vaccines play in feline health, and understanding cat owner concerns and preferences, veterinarians have an opportunity to improve feline vaccination compliance.

Veterinary Sciences doi: 10.3390/vetsci13070645

Authors: Andrés Montesinos Barceló Alicia Tortosa García María Ardiaca García

Background: Hyperuricemia is a common problem in captive reptiles and is associated with gout and renal disease. Rasburicase is a recombinant urate oxidase that converts uric acid into allantoin, a more water-soluble metabolite. This study evaluated the safety and efficacy of rasburicase in clinically healthy ball pythons (Python regius). Methods: Eight adult ball pythons were included in a prospective crossover study. Six experimental conditions were evaluated: fasting with and without rasburicase (0.2 mg/kg IM), feeding with a day-old chick with and without rasburicase, and feeding with a mouse with and without rasburicase. Plasma uric acid concentrations were measured at 0, 24, 48, 72, and 96 h. Results: Rasburicase did not significantly affect fasting uric acid concentrations. Feeding increased plasma uric acid concentrations, with peak values occurring between 24 and 72 h. Following chick consumption, rasburicase promoted a faster decline in UA concentrations, although differences were not significant (p = 0.907). Following mouse consumption, rasburicase significantly reduced UA concentrations (p = 0.002), with a mean reduction of 37.85%. No changes were observed in behavior, appetite or defecation. Conclusions: Rasburicase was well tolerated and reduced postprandial uric acid concentrations in ball pythons.

Veterinary Sciences doi: 10.3390/vetsci13070644

Authors: Anna Didkowska Victor Martín Santander Daniel Klich Michal Skibniewski Katarzyna Matusik Marlena Wojciechowska Wanda Olech Krysztof Anusz Diana Marteles-Aragüés Sergio Villanueva-Saz Antonio Fernández

Knowledge of the reference intervals for serum proteins of the European bison (Bison bonasus) can be of great help in the diagnosis of infectious and inflammatory diseases, which are very common in these animals; however, reference intervals for this species are unknown. A total of 131 serum samples were obtained from the European bison from different locations in Poland. Agarose gel electrophoresis (AGE) and capillary zone electrophoresis (CZE) were used to separate and identify protein fractions. In all sera, six fractions were identified: albumin, α1, α2, β1-, β2-, and γ-globulins by both analytical methods. Reference intervals (90% CI) were determined. Comparisons were made to find significant differences based on the sex, age or lifestyle of the animals. Concordance between the methods was assessed using Passing–Bablok regression, Bland–Altman analysis, and Lin’s concordance correlation. A higher percentage of α1-globulins was found in the females than in the males. Captive bison had significantly lower γ-globulin levels compared to free-living bison. The γ-globulin levels increased with the age of the animals. The Bland–Altman analysis demonstrated a systematic bias indicating no good agreement between methods, and only the values for albumin, β1-, β2-, γ-globulins, and total globulins were considered acceptable according to Lin’s concordance correlation. Both methods are adequate for identifying serum proteins, but they are not interchangeable, necessitating the use of separate reference intervals for each method.

Veterinary Sciences doi: 10.3390/vetsci13070643

Authors: Adrian Stancu Janos Degi Iasmina Luca Diana Maria Degi Simona Marc Sorin Aurelian Pașca Sorin Octavian Voia Adela Marcu

Canine parvovirus type 2 (CPV-2) remains an important cause of morbidity and mortality in young dogs worldwide. Although the enteric form of the disease is well characterized, myocardial involvement associated with CPV-2 infection remains incompletely understood. The present retrospective study aimed to investigate myocardial lesions in young dogs and to evaluate their association with CPV-2 DNA detection using histopathological examination and PCR-based molecular analysis of formalin-fixed paraffin-embedded (FFPE) cardiac tissue. A total of 27 dogs aged between 3 and 11 months, submitted for diagnostic necropsy between 2019 and 2021, were included in the study. Histopathological evaluation was performed on myocardial tissue sections stained with hematoxylin and eosin, while conventional PCR targeting the VP1/VP2 region was used for the detection of CPV-2 DNA. Associations between PCR positivity and histopathological findings were assessed using Fisher’s exact test. Myocardial abnormalities were identified in all examined cases (27/27, 100%), with myocarditis consistently observed throughout the study population. Cardiomyocyte necrosis and myocardial fibrosis were identified in 9/27 (33.3%) and 14/27 (51.9%) cases, respectively. CPV-2 DNA was detected in 9/27 myocardial tissue samples (33.3%). Cardiomyocyte necrosis was significantly more frequent in CPV-2-positive dogs than in PCR-negative animals (77.8% vs. 11.1%; p ≈ 0.001). Similarly, myocardial fibrosis was identified more frequently in CPV-2-positive cases (100.0% vs. 27.8%; p ≈ 0.001). Myocarditis was present in all examined animals and therefore could not be evaluated statistically according to PCR status. The detection of CPV-2 DNA in myocardial tissues exhibiting histopathological lesions supports a possible association between CPV-2 infection and myocardial injury in young dogs. However, the retrospective design, limited sample size, and the use of conventional PCR do not allow conclusions regarding causality or direct viral involvement in lesion development. Further studies incorporating larger case series and complementary diagnostic techniques are needed to clarify the pathogenesis and clinical significance of CPV-2-associated myocardial lesions.

Veterinary Sciences doi: 10.3390/vetsci13070642

Authors: Sandra V. Kielmann Roswitha Merle Jörg R. Aschenbach Annika Fels Katharina Charlotte Jensen

Extramural clinical rotations (ECR) are an integral part of veterinary education in Germany and give students the opportunity to practice and deepen their knowledge and skills. In this study, students from all five veterinary schools in Germany who completed their practical year between the years of 2022 and 2025 were surveyed online regarding their perceived preparedness, their evaluation of teaching during ECR, and what aspects were relevant to them when choosing ECR. Data records of 386 students were analyzed. Being assigned a direct supervisor and feedback meetings were among the five most important aspects for students when choosing their ECR, underlining the importance of a clear structure and constructive feedback for successful learning. More than half of the students reported feeling insufficiently prepared, both theoretically and practically, for surgery. Although the students already passed their propaedeutics exams years before, only 46% of respondents stated having felt (very) well prepared in terms of propaedeutics knowledge going into their ECR, 59% felt (very) well prepared in terms of performing general examinations (routine clinical examinations) and 20% in terms of specific examinations (e.g., neurological or rectal examinations). Meanwhile, 75% of students rated the quality of practical teaching during their ECR positively. However, in some areas—such as specific examinations and diagnostic imaging—there was still room for improvement. Overall, the results suggest that ECR are generally perceived positively. Concerning the feeling of mostly insufficient preparation, further studies are needed to elaborate whether teaching should focus more on practical skills.

Veterinary Sciences doi: 10.3390/vetsci13070641

Authors: Ali Mujtaba Shah Dongxu Xia Wence Wang Yuan Yuan Ali Raza Shah Ali Mustafa Shah Nazir Ahmed Khan Weijie Pan Wei Shi Guoqiang Chen Fu Yang Hongxia Zhao Qingyun Cao

Palm fruits are produced extensively in tropical and subtropical regions and consumed worldwide. However, over 20% of the total yield is discarded due to inferior quality, resulting in significant agricultural waste and economic loss. To mitigate this challenge and enable the safe valorization of discarded dates (DD) in animal feeding systems, this study employed solid-state fermentation (SSF) to upgrade the nutritional quality of DD and evaluated its potential as a functional feed ingredient for goats. Twenty-four male goats (6 months old; initial body weight 25.86 ± 0.25 kg) were randomly assigned to one of three dietary treatments: a basal diet (control), a diet containing 10% raw DD (D1), and a diet containing 10% solid-state fermented DD (D2). Inclusion of DD in the diet significantly increased average daily gain (ADG), final body weight (BW), and feed efficiency, with the highest values recorded for D2 (p < 0.05). Feeding of DD altered (p < 0.001) all measured rumen fermentation parameters, except pH, with higher levels (p < 0.05) of total volatile fatty acids, propionate, microbial crude protein, and ammonia nitrogen recorded for D1 and D2, as compared to control. Similarly, blood biochemistry revealed elevated total protein, albumin, and globulin in both supplemented groups (p < 0.05), whereas higher glucose and cholesterol levels were recorded for the D1 group (p < 0.05). Notably, systemic antioxidant status improved with the inclusion of SSF fermented DD, as evidenced by increased superoxide dismutase, glutathione peroxidase, and catalase activities, alongside reduced malondialdehyde levels (p < 0.05). The inclusion of DD in the diet decreased cooking and drip losses, and decreased shear force (indicating enhanced tenderness) and water-holding capacity (p < 0.05), with better values recorded for D2. Carcass protein and fat contents increased with the inclusion of DD in the diet, with higher values recorded for D2 (p < 0.05). Fatty acid analysis revealed higher (p < 0.05) contents of rumenic acid and octadecenoic acid in D2, as compared to D1 and control. The concentrations of lysine, methionine, threonine, leucine, and valine in meat were also higher in D2-fed goats (p < 0.05). In conclusion, incorporating solid-state-fermented discarded dates into goat diets represents a promising and sustainable strategy to valorize agricultural waste while concurrently improving growth performance, antioxidant status, meat quality, and selected nutrient profiles of goat meat. These preliminary findings warrant validation in larger-scale production.

Veterinary Sciences doi: 10.3390/vetsci13070640

Authors: Aya S. Elmasry Eman A. Elwakeel Ali M. Allam Marwa F. A. Attia Alaa. T. Elmaria Elsayed. E. M. Badr Sobhy M. A. Sallam

Mastitis is a major and costly dairy disease that reduces milk yield and quality and harms animal welfare. This study evaluated infrared thermography (IRT) combined with machine learning (ML) for non-invasive mastitis screening in dairy cows and explored links with biological and feeding-system variables in Egyptian farms. A total of 976 thermal udder images obtained from 488 Holstein cows were used, including 708 healthy and 268 mastitic images. Images were captured before milking, processed with CLAHE, resized to 224 × 224 pixels, and split using cow-level grouping before augmentation to prevent animal-level data leakage. The training set contained 780 original images and was augmented to a balanced 4708-image set (2354 per class), while the held-out test set remained unaugmented, with 196 original images (142 healthy and 54 mastitic). EfficientNetB3 with global average and max pooling extracted 3072 thermal features, and ten ML classifiers were evaluated. In the image-level hold-out evaluation, MLP achieved the best performance (accuracy = 86.22%, AUC = 0.9184, sensitivity = 74.07%, specificity = 90.85%), followed by SVM (accuracy = 83.67%, AUC = 0.8963). A separate group-based five-fold cross-validation yielded a more conservative AUC of 0.6812 ± 0.1323 and accuracy of 0.6244 ± 0.0642. Logistic regression analyses did not identify statistically significant associations between model predictions and somatic cell count (SCC), California Mastitis Test (CMT), blood biomarkers, or nutritional variables at p < 0.05. Ration A (Delta Misr) showed a higher observed mastitis incidence (20/40; 50.0%) than Ration B (Copenhagen; 16/45; 35.6%), but nutritional predictors were not statistically significant, indicating that farm-level confounding should be considered. Overall, IRT with ML remains a promising non-invasive screening approach, but broader multicenter datasets and independent external validation are needed before routine farm deployment.

Veterinary Sciences doi: 10.3390/vetsci13070639

Authors: Ming Liu Wenhui Zhang Wanyu Zhang Patrick Butaye Zhiqiang Wang Ruichao Li

Vibrio fluvialis is an important zoonotic pathogen that poses a growing threat to both public health and aquaculture, particularly to shrimp and fish farming. In this study, we report that a seafood-associated V. fluvialis strain, 10M-VF, possesses two genomic islands, SGI1-VfNDM1 and GIflu-1, in addition to hemolysins and type VI secretion systems (T6SSs). SGI1-VfNDM1 is a new member of the Salmonella genomic island 1 (SGI1) and harbors several antibiotic resistance genes. A potential transferable region, ISCR1-trpF-bleMBL-blaNDM-1, was observed in SGI1-VfNDM1, suggesting that ISCR1 may have mediated the integration of blaNDM-1 into this island. Additionally, GIflu-1 represents a novel resistance island. Other GIflu variants carrying resistance genes in Vibrio all possess ISCR2, but their resistance regions vary. This indicates that ISCR2 may contribute to the integration of antibiotic resistance genes into GIflu islands. Notably, not every GIflu variant carries drug resistance genes. To the best of our knowledge, GIflu-1 is a previously unreported resistance island, and no previous report has documented the co-existence of two distinct resistance genomic islands in a single Vibrio isolate. The emergence of such resistance islands in a seafood-borne pathogen raises concerns for the maintenance and potential dissemination of clinically important resistance genes in aquatic food systems and underscores the need for ongoing surveillance of resistance determinants in seafood-borne Vibrio populations from a One Health perspective.

Veterinary Sciences doi: 10.3390/vetsci13070638

Authors: Weldeab Solomon Ghebrezgabher Jiazhen Ge Guodong Song Fuying Zheng Yuefeng Chu

Brucellosis remains a critical zoonotic threat to public health and livestock economies globally, particularly constraining small ruminant productivity in Africa, yet continental data remain scarce. The objectives of this review are to provide apparent small ruminant brucellosis prevalence/positivity estimates across Africa by country, region, continent, and diagnostic modality. A systematic search was conducted on PubMed, ScienceDirect, and Google Scholar for articles published from 2000 to 2025. Data from 151 eligible studies representing 28 countries were extracted and meta-analyzed. Due to extreme heterogeneity (I2 > 97%), a single continental pooled estimate is not epidemiologically meaningful. For descriptive reference, Africa’s apparent pooled seroprevalence was 4.9% (95% CI: 3.0–7.5%; prediction interval: 0.1–32.1%), and pooled PCR positivity was 12.7% (95% CI: 6.0–21.9%; prediction interval: 0.8–38.9%). Descriptive regional seroprevalence ranged from 3.3% (95% CI: 1.2–7.4%) (West Africa) to 6.2% (95% CI: 3.8–9.4%) (East Africa). Country-level crude seroprevalence ranged from 0.18% (Morocco) to 19.53% (Libya), with diagnostic method prevalence ranging from 3.37% (2-Mercaptoethanol (2-ME)) to 21.54% (Whole Genome Sequencing (WGS)). Both B. abortus and B. melitensis are reported with significant variations. Data absence from several African countries highlights a critical knowledge gap. We propose multi-level prevention and control strategies.

Veterinary Sciences doi: 10.3390/vetsci13070637

Authors: Yunfei Zhai Jiaxuan Song Hantong Weng Haihui Wang Tianqin Hu Zhaoyu Han

Nitrogen use efficiency (NUE) is commonly calculated as the ratio of milk nitrogen output to nitrogen intake in dairy cows. However, because milk nitrogen output is intrinsically determined by milk production and nitrogen intake is largely driven by dry matter intake, conventional NUE is mathematically dependent on production level and feed intake. This dependency makes it difficult to distinguish apparent efficiency caused by higher milk yield from intrinsic biological efficiency in nitrogen utilization. Therefore, this study aimed to evaluate NUE in dairy cows using a combined mathematical and metabolic framework based on residual NUE (rNUE) analysis. A total of 126 early-lactation Chinese Holstein cows were screened, and 16 high-NUE and 16 low-NUE cows were selected after matching for parity, days in milk, and dry matter intake. High-NUE cows had greater milk yield, milk nitrogen output, and NUE than low-NUE cows, despite similar nitrogen intake. They also exhibited higher ruminal microbial crude protein and ammonia nitrogen concentrations, a lower acetate-to-propionate ratio, and reduced circulating essential and total amino acid concentrations. Multiple regression analysis showed that energy-corrected milk and dry matter intake explained 71.4% of the variation in NUE. The residuals from this model were defined as rNUE, which was independent of milk production. After removing production-related effects, rNUE remained positively associated with ruminal microbial crude protein and ammonia nitrogen concentrations, and negatively associated with the acetate-to-propionate ratio and circulating amino acid pools. These findings indicate that conventional NUE in dairy cows is largely driven by production level, whereas residual-based modeling can identify a production-independent component associated with rumen nitrogen metabolism and amino acid utilization. The residual NUE approach provides a useful mathematical and metabolic framework for evaluating intrinsic NUE in dairy cows beyond milk production level.

Veterinary Sciences doi: 10.3390/vetsci13070636

Authors: Zhihua Wang Te Pi Yanling Ren Cuiping Wang Yishan Li Feng Li Shouqing Yan

The Wadi sheep (WAD) is a unique Chinese indigenous breed previously reported to show adaptation to humid and saline–alkali lowlands, disease resistance, high fecundity, and favorable meat quality. However, the recent widespread introduction of commercial sheep breeds has led to a sharp decline in the purebred WAD population, posing serious threats to the preservation of its unique genetic resources. Therefore, there is an urgent need to assess the current population structure and genetic diversity of WAD to support its conservation and sustainable utilization. In this study, we systematically analyzed genomic variation, population structure, and selection signatures using whole-genome resequencing data from 30 WAD sheep and 80 publicly available genomes representing five other breeds obtained from the NCBI database. Population genomic analyses revealed that WAD retains substantial genetic diversity and exhibits a distinct population structure, shaped by its unique breeding history. Selection signature analyses using FST, π ratio, and XP-EHH identified 457 candidate genes under positive selection, which are associated with key biological processes including environmental adaptation, immune defense, muscle traits, growth, and reproduction. These findings identify genomic regions potentially related to reported WAD traits and provide a theoretical basis for its conservation, molecular breeding, and sustainable utilization in lowland environments.

Veterinary Sciences doi: 10.3390/vetsci13070635

Authors: Abdul Qadeer Mohamed Tharwat Ibrahim F. Halawani Fuad M. Alzahrani Khalid J. Alzahrani Fahad A. Alshanbari Muhammad Zahoor Khan

Endometritis—inflammation of the endometrial lining—imposes a substantial reproductive and economic burden in both human gynecology and livestock production, where it is a leading cause of recurrent implantation failure in humans and the costliest reproductive disorder in cattle. Conventional management is overwhelmingly antibiotic-based, yet escalating antimicrobial resistance, tissue and milk residues, microbiota disruption and high relapse rates have eroded its efficacy and acceptability, creating an urgent need for mechanism-based, host-directed alternatives. Here we synthesize the expanding evidence positioning dietary flavonoids and traditional Chinese medicine (TCM) formulations as such interventions. Across diverse compounds and preparations, anti-endometriotic activity converges on a tractable set of molecular nodes: TLR4/NF-κB signaling, the NLRP3 inflammasome–pyroptosis axis, the Keap1/Nrf2/HO-1 antioxidant program, PI3K/AKT and PPAR-γ signaling, ferroptosis, and the gut–uterus microbial–metabolite axis. Veterinary field studies report cure rates and fertility outcomes rivaling first-line antibiotics, while integrative case reports show benefit in antibiotic-refractory human chronic endometritis. Translation remains constrained by poor bioavailability, formulation heterogeneity, over-reliance on lipopolysaccharide-only models and a scarcity of randomized trials—barriers now addressable through nanocarrier delivery, network-pharmacology-guided standardization and biomarker-stratified designs. Flavonoids and TCM are best viewed not as substitutes for antibiotics but as a mechanistically rational, multi-target strategy aligned with One Health antimicrobial stewardship.

Veterinary Sciences doi: 10.3390/vetsci13070634

Authors: Carmen G. Pérez-Santana Sara Cazorla-Rivero Enrique Rodríguez Grau-Bassas Bernardino Clavo Francisco Rodríguez-Esparragón

Primary hepatic masses in dogs represent a heterogeneous group of lesions with variable biological behavior and challenging preoperative characterization. The objective of this retrospective study was to describe the clinical presentation, diagnostic findings, surgical management, and outcome of dogs with primary hepatic lesions treated surgically. Ten dogs with resectable hepatic lesions and no evidence of extrahepatic metastasis were included. Clinical records, imaging findings, histopathological diagnoses, treatment, and follow-up data were reviewed. Histopathological diagnoses included hepatocellular carcinoma (n = 3), nodular hyperplasia (n = 2), lobular hyperplasia (n = 1), hepatocellular adenoma (n = 1), undifferentiated sarcoma (n = 1), osteosarcoma (n = 1), and one case without a definitive histological diagnosis. Tumor size ranged from 3.3 to 18 cm. Both benign and malignant lesions were represented across this size range. Cytological findings were discordant with the final histopathological diagnosis in three dogs, whereas one sample was non-diagnostic due to blood contamination. Abdominal ultrasound identified solitary lesions in all evaluated dogs, although surgery revealed previously undetected multifocal disease in two cases. Six of the ten lesions involved the right medial hepatic lobe, although the small sample size precludes conclusions regarding lesion distribution. All dogs underwent surgical resection. Two perioperative deaths occurred secondary to postoperative renal failure. Among the surviving dogs, clinical status during follow-up was generally reported as improved compared with preoperative presentation, and survival times ranged from 4 to more than 730 days, including dogs with malignant neoplasms. Four dogs remained alive and disease-free at the end of the follow-up period (>730 days). This retrospective case series illustrates the clinical and pathological heterogeneity of primary hepatic masses in dogs. Discrepancies between preoperative assessment and intraoperative or histopathological findings were observed in some cases. Given the small sample size, heterogeneous diagnoses and non-standardized diagnostic workup, these observations should be interpreted cautiously and considered hypothesis-generating.

Veterinary Sciences doi: 10.3390/vetsci13070633

Authors: Nourhan Nassar Mohamed Tharwat Aya Tayel Muhammad Tariq Yasir Muhammad Khan Fahad A. Alshanbari Ibrar Muhammad Khan

The gut microbiome plays a central role in regulating nutrient utilization, immune function, and disease resistance, thereby directly influencing growth performance and feed efficiency. Existing microbiome modulation strategies, including probiotics, prebiotics, dietary interventions, and antibiotic alternatives, are critically evaluated. Despite their reported benefits, the effectiveness of these approaches often remains inconsistent across production systems. Evidence suggests that this variability is largely driven by complex interactions among microbial communities, host factors, and environmental and management conditions, which are frequently overlooked in conventional intervention-based approaches. To address this gap, this review proposes an integrated microbiome–host–environment framework that links microbial ecology with host physiology and production conditions. The framework provides a systems-level perspective for understanding the factors governing microbiome stability and production responses, offering a basis for more targeted and reliable microbiome management strategies. Finally, current challenges and future research priorities are discussed, including the integration of multi-omics technologies, precision nutrition, and data-driven approaches to support next-generation poultry production systems. By emphasizing the interconnected nature of microbiome regulation, this review contributes a conceptual foundation for improving broiler productivity and sustainability through more consistent and effective microbiome optimization.

Veterinary Sciences doi: 10.3390/vetsci13070632

Authors: Li Li Lu Zhang Chunyu Xue Leimin Zhang Dongyu Huang Mingchun Ren Haifeng Mi Hualiang Liang

Malondialdehyde (MDA) is a common oxidation product in deteriorated aquatic feed, which easily induces oxidative damage and quality deterioration in farmed fish, yet its systemic effects on the physiological function and body color of channel catfish (Ictalurus punctatus) remain poorly understood. This study aimed to investigate the influences of dietary exogenous MDA on antioxidant capacity, immune function and body coloration of channel catfish. A 30-day feeding trial was carried out with four dietary MDA levels of 0, 22.3, 44.6 and 66.8 mg/kg. Relevant physiological and pigment indices of skin and muscle were determined. Exogenous MDA significantly increased plasma transaminase levels and tissue MDA content, decreased antioxidant enzyme activities, and altered the transcription of antioxidant and immune-related genes in skin and muscle, causing oxidative stress and immune dysfunction. It also suppressed tyrosinase activity, downregulated melanin-synthesis-related genes, reduced melanin deposition and promoted uranidin accumulation in the two tissues. Collectively, dietary exogenous MDA impairs antioxidant and immune performance, disrupts pigment metabolism, and ultimately leads to body yellowing in channel catfish.

Veterinary Sciences doi: 10.3390/vetsci13070631

Authors: Shuo Zhao Xianhua Wu Ying He Jinmu Lin Xinlin Zhong Baojiang Lin Wen Zhao Xinting Xu Qunpeng Duan Xunye Yang Han Shao Ying Peng Yilan Xu Tingting Chen Chenyu Quan Bingxia Lu Wenfeng Wang Yang Qin Zhongwei Chen Yangqing Lu Yibin Qin

Porcine rotavirus (PoRV) has emerged as a primary pathogen causing viral diarrhea in pigs, resulting in significant economic losses. This study was conducted to systematically characterize the epidemiology and genotypic characteristics of PoRV in Guangxi, China. A total of 870 diarrheic pig samples were collected from Guangxi during 2020–2025. The qRT-PCR results indicated an overall PoRV-positive rate of 41.38% (360/870), and the annual positivity rate showed an overall upward trend. The genetic evolutionary analysis of the VP4, VP6, and VP7 genes indicated that PoRV predominantly belonged to the A group and the predominant P genotype observed was P[13] (76.83%), while the G genotypes were G5 (36.56%) and G9 (33.33%). The most prevalent genotype combinations were G9P[13]I5 and G5P[13]I5. CH-GXGL-PoRV-3151-2021, a PoRV strain isolated from positive samples, was identified via RT-PCR, qRT-PCR, whole-genome sequencing, and IFA. This strain was assigned the 11-segment genotype constellation G9-P[23]-I5-R1-C1-M1-A8-N1-T1-E1-H1 based on whole-genome sequencing. NSP1 and NSP2 showed high similarity to human rotavirus strains, whereas VP1–VP4, VP6, VP7, and NSP3–NSP5 showed high similarity to porcine rotavirus strains. This study indicates the widespread circulation of PoRV in Guangxi, with multiple G genotypes, including G9, G5, G4, G3, G2, and G26, being detected. The isolated G9P[23]I5 strain exhibits the same genotype as the strains that have become increasingly prevalent in recent years. This strain may represent a possible reassortant between porcine and human rotaviruses. This study offers significant insights into the epidemiology of PoRV and the prevalent genotypes in Guangxi, thereby supporting the development of targeted prevention strategies and novel vaccines.

Veterinary Sciences doi: 10.3390/vetsci13070630

Authors: Zarroug Hassan Ibrahim

The second stomach compartment (C2) of the dromedary camel (Camelus dromedarius) plays an important role in digestion. However, detailed morphological and histochemical data remain limited. This study aimed to investigate the gross anatomy, histological organization, histometric features, and histochemical distribution of muco-substances in C2. The study was conducted on twenty dromedary camels, including fetuses and adults. Gross anatomical observations were performed on eight fresh and fixed specimens, while histological, histometric, and histochemical analyses were carried out on samples from twelve adult camels using routine and special staining techniques to identify neutral and acidic mucins. C2 was the smallest gastric compartment, located on the right side of the abdominal cavity and partially continuous with C1. Its mucosa formed chambered zones supported by prominent longitudinal muscular bands. Histologically, C2 comprised glandular and non-glandular regions. The glandular mucosa contained gastric pits and branched tubular glands with mucous, chief, and parietal cells, whereas the non-glandular region was lined by keratinized stratified squamous epithelium. Submucosal lymphoid aggregations were observed near the C2–C3 junction. Histometric analysis revealed a markedly developed tunica muscularis. Strong PAS and Alcian blue reactions indicated abundant neutral and acidic mucins. These findings demonstrate that C2 is a structurally specialized compartment supporting digestion, mucosal immune defense, and adaptation to arid environments, clearly distinguishing it from the reticulum of true ruminants.

Veterinary Sciences doi: 10.3390/vetsci13070629

Authors: Rasha S. Sakla Aida A. Abd El-Wahed Wael Mahmoud Aboulthana Sobhia S. Sayed

Nutrition is one of the main challenges facing honey bees, especially during autumn and winter. This work aimed to evaluate the effects of feeding Apis mellifera with a protein-enriched diet (YCPC) compared to a control diet. Development of the hypopharyngeal glands (HPGs), dimensions of the second wax mirror, and the stinging apparatus were measured in the two groups. Additionally, key biochemical parameters were assessed in homogenized worker honey bees. No significant differences were observed in hypopharyngeal gland (HPG), acini diameters or surface area, or in the measurement of the stinging apparatus and poison sac, except for a slight decrease in the newly emerged bees. YCPC supplementation significantly elevated total soluble protein content in newly emerged and nurse bees. Lower lipid levels were observed in newly emerged and nurse bees from the supplemented group and higher lipid reserves in were observed in supplemented foragers. No significant variation was observed in total carbohydrate content, except for a significant reduction in supplemented newly emerged bees. Acetyl cholinesterase activity was higher in younger bees and lower in foragers. A slight elevation in α and β-esterase expression was detected in the supplemented group, supporting enhanced detoxification capacity. YCPC enhances the biochemical capacity of honey bees, particularly in the early adult stages. However, glandular and morphological development were not affected.

Veterinary Sciences doi: 10.3390/vetsci13070627

Authors: Indra Sara Klumb Axel Wehrend Abbas Farshad

Background: Accurate assessment of sperm morphology is essential for evaluating the quality of cryopreserved canine semen used in artificial insemination and for improving cryopreservation protocols. This study compared six staining techniques, Eosin, Eosin–Nigrosin, Diff-Quick®, Hemacolor®, Spermac®, and Formol-citrate Bengal Rose, for light-microscopic evaluation of frozen–thawed canine spermatozoa. Methods: Semen from ten dogs was thawed, divided into four aliquots, and either left untreated or exposed to thermal stress at 6 °C, 18 °C, or 37 °C for two hours to induce morphological variation. A total of 360 slides and 960 evaluations were performed immediately after staining and again after 24 h, 7 days, and 3 months to assess staining quality and stability over time. Results: Eosin produced stable staining for up to three months and was the most economical method, though its initial detail recognition was lower than that of Spermac® and Formol-citrate Bengal Rose. Eosin–Nigrosin showed reduced contrast and detail. Diff-Quick® provided better contrast than Eosin–Nigrosin, while Hemacolor® maintained consistent quality regardless of stress treatment or storage duration. Spermac® yielded the highest initial morphological detail but deteriorated during storage. Formol-citrate Bengal Rose combined high detail recognition with stable staining throughout the study. Cryopreservation increased looped tails, and incubation at 37 °C markedly elevated pathological sperm and rudimentary tails. Conclusions: All six staining methods were suitable for evaluating and archiving frozen–thawed canine semen. Formol-citrate Bengal Rose and Spermac® offered the best detail, while Eosin provided a cost-effective option with excellent long-term stability.

Veterinary Sciences doi: 10.3390/vetsci13070628

Authors: Shang Jiang Haihua Feng Chao Wang Xiliang Du Lin Lei Wenwen Gao Guowen Liu Xinwei Li Yuxiang Song

Heat stress severely impairs immune function and threatens dairy cow health and productivity. Although heat stress suppresses bovine neutrophil (PMN) phagocytosis, its effects governing PMN survival remain unclear. L-arginine (L-Arg) exerts immunomodulatory and cytoprotective effects, yet its role in repairing heat-damaged PMN has not been defined. In this study, an in vitro heat stress model (42 °C for 2.5 h) was established. Heat stress significantly reduced cell viability, induced abnormal nuclear morphology, and triggered apoptotic signaling, accompanied by severe oxidative stress (elevated reactive oxygen species (ROS)/Malondialdehyde (MDA), decreased SOD/glutathione peroxidase (GSH-Px)). The ROS scavenger N-Acetylcysteine (NAC) mitigated both oxidative stress and apoptosis, confirming oxidative stress as a core mediator of PMN apoptosis induced by heat stress. Pretreatment with 4 mmol/L L-Arg significantly alleviated heat-induced oxidative damage and apoptosis. Mechanistically, in bovine PMN, we first demonstrated that heat stress blocks nuclear factor erythroid 2-related factor 2 (NFE2L2) nuclear translocation; inhibition of NFE2L2 (ML385) abolished L-Arg’s protection, verifying the NFE2L2-dependent ROS scavenging pathway. In conclusion, this study reveals that heat stress induces bovine PMN oxidative injury and apoptosis by suppressing NFE2L2-mediated ROS scavenging, and L-Arg restores PMN viability and immune resilience by reactivating the NFE2L2 antioxidant pathway. These findings provide a targeted L-Arg supplement strategy to improve PMN survival and disease resistance in heat-stressed dairy cows.

Veterinary Sciences doi: 10.3390/vetsci13070626

Authors: Xianfeng Hui Shihuan Ding Shuoxiang Gao Shuochen Xu Tiesuo Zhao Xiaowei Tian Hui Wang

Influenza viruses continue to pose a significant threat to both animal and public health due to their rapid evolution and the frequent emergence of antiviral resistance. Host-directed antiviral (HDA) strategies, which target host factors essential for viral replication, may represent an alternative to conventional virus-targeting approaches. However, the identification of reliable and therapeutically actionable host targets remains a major challenge, primarily due to the complexity and context dependency of host–virus interactions. Recent advancements in multi-omics technologies, including functional genomics, transcriptomics, and proteomics, have facilitated the systematic characterization of host factors involved in influenza virus infection. These methodologies have unveiled intricate regulatory networks that govern viral replication and host immune responses. Nonetheless, translating large-scale datasets into biologically meaningful targets necessitates robust integrative frameworks. In this context, artificial intelligence (AI) and machine learning methods offer powerful tools for data integration, target prioritization, and predictive modeling. In this Review, we summarize current insights into host factors that regulate influenza virus infection and discuss how multi-omics and AI-driven approaches are expediting host target discovery. Furthermore, we highlight the potential of these strategies to enhance antiviral development while addressing key challenges related to specificity, safety, and translational application. Collectively, these advancements lay a foundation that may support the rational design of next-generation host-directed antivirals.

Veterinary Sciences doi: 10.3390/vetsci13070624

Authors: Cristian-Olimpiu Martonos Cristian Constantin Dezdrobitu Little William Brady Laszlo-Andras Nagy David Hilchie Pompei Bolfa Alexandru Ion Gudea

This study examines the auditory ossicles of 55–56-day gestational age dog fetuses and shows that the basic three-bone middle ear pattern is already present, but the bones remain partly cartilaginous and are actively ossifying. The tympanic cavity contained a gelatinous, mesenchyme-like material, making dissection difficult. The malleus was the largest and most laterally located ossicle, with an oval head, short neck, curved manubrium, and notable variability in the anterior process. The incus had a rectangular body with short and long processes of similar length, and the lenticular process plus a synovial incudo-malleal joint were already present. The stapes was the smallest ossicle, triangular in shape, with a shorter posterior crus and early ossification in the distal crura and footplate. Histology showed active bone formation with medullary cavities, trabeculae, and osteoclast activity in specific regions, especially the malleal neck, the incudal long process, and the distal stapes, as the metrical data may reflect early mechanical specialization. Overall, the paper concludes that fetal dogs already possess the mammalian ossicular plan, but the structures remain immature, reflecting a clear histological evolution from cartilage-dominant tissue toward organized bone.

Veterinary Sciences doi: 10.3390/vetsci13070625

Authors: Yuqing He Monan Chen Chaohao Yu Xin Wang Xinyang Li Wei Guo

The epidemiology of the nematode Eustrongylides sp. remains poorly understood in natural fish populations, especially under alpine conditions. This study investigated its infection dynamics in Triplophysa strauchii, an endemic fish in an alpine wetland in northwestern China. A total of 720 fish were sampled monthly from April to November 2025. Parasitic larvae were identified using ribosomal ITS sequences. Hurdle models were employed to assess infection risk and intensity, incorporating host length, sex, and month as predictors. The overall prevalence was 21.8%, with a mean intensity of 4.36 ± 6.83 worms per infected fish. Infection dynamics showed two seasonal peaks: a spring peak with high intensity and a late-summer peak with moderate prevalence but low intensity. Host length was the most stable predictor for both infection risk (OR = 1.916 per cm, 95% CI: 1.232–2.977) and intensity, with significant length × month interactions (length × May: IRR = 4.847, p = 0.005; length × November: OR = 0.418, p = 0.005), indicating seasonal modulation of the length effect. These findings provide foundational insights into parasite transmission in alpine wetlands and highlight public health concerns regarding fish consumption.

Veterinary Sciences doi: 10.3390/vetsci13070623

Authors: Mario Baratta Paolo Accornero Silvia Miretti Eugenio Martignani

This review examines the relationship between climate-driven heat stress (HS) and bovine mastitis in the Po Valley, a key European dairy region characterized by intensive production systems and increasing climatic vulnerability. It aims to contextualize how rising temperature–humidity index (THI) levels influence animal health and productivity. This study synthesizes the current literature on biometeorological conditions, epidemiological trends, and physiological mechanisms linking HS to mastitis. Evidence indicates that prolonged exposure to elevated THI impairs thermoregulation, disrupts endocrine and metabolic balance, and weakens immune function, thereby increasing susceptibility to intramammary infections. Epidemiological data reveal a clear seasonal pattern, with mastitis incidence peaking during summer months and a growing predominance of environmental pathogens. Additionally, HS negatively affects milk yield and quality, amplifying economic losses in dairy systems. The findings highlight that mastitis in this context is not merely an infectious disease but a multifactorial condition shaped by environmental, physiological, and management factors. Overall, this review underscores the need for integrated mitigation strategies, including improved housing, nutrition, genetic selection, and precision monitoring, to enhance resilience. In the face of ongoing climate change, adapting dairy production systems will be essential to safeguard animal welfare, maintain productivity, and ensure the long-term sustainability of the Po Valley dairy sector.

Veterinary Sciences doi: 10.3390/vetsci13070622

Authors: Pedro Antonio Robles-Trillo Christopher D. Lu Luis Jesús Barrera-Flores Rafael Rodríguez-Venegas Martín Alfredo Legarreta-González Rafael Rodríguez-Martínez

Attempting to detect and improve the management of Ketosis, the objective of this study was to determine and confirm the relationship between hours of activity, rumination time, conductivity, and milk production with the presence of ketosis in cows during the transition period in dairy cows in the Comarca Lagunera region, the heart of the dairy cattle production in Mexico. Data were collected in a large scale dairy cattle study. High-precision electronic collar sensors, high-precision electronic scales, and online electronic weighing sensors were employed to determine activity and ruminating time, milk electrical conductivity, and milk yield, respectively. All data were collected and integrated using an electronic peripheral management and control software. Using urinary ketone bodies measured by qualitative strips as the biomarker for ketosis, 10.50% of the cows were found to be positive for ketosis, while the remaining 89.50% were negative. The mean and standard error for activity time (AT), ruminating time (RT), milk electrical conductivity (CE) and milk yield (MY) in normal (N) vs ketotic (P) cows were: AT N 61.38, ± 0.39, AT P 39.08 ± 0.49; RT N 530.85 ± 2.94, RT P 295.24 ± 10.69; CE N 5.68 ± 0.03, CE P 9.13 ± 0.11; and MY N 38.87 ± 0.29, MY P 20.34 ± 0.54. Exploratory Factor Analysis (EFA) was conducted for the purpose of uncovering the underlying structure of the data by identifying latent constructs that influence the observed variables. The EFA estimated two factors which explained 62% of the variation observed. The Factor 1 (MR1) comprising the variables MY and EC, and Factor 2 (MR2), which consists the variables AT and RT. High-precision measurement sensors along multivariable analyses could facilitate the establishment of a correlation between ketosis and variables associated with the physiology, well-being, and productivity of bovines in the transition period. It further open the possibility of early detection of metabolic diseases such as ketosis.

Veterinary Sciences doi: 10.3390/vetsci13070621

Authors: Liyu Yang Jiaxin Liu Xuanxuan Zhang Yake Wang Shufan Liu Chenxi Ling Xinfeng Li Kun Liu Yong Huo Guangwei Zhao Qiuliang Xu Hongyu Deng Congcong Li

Aflatoxin B1 (AFB1) causes acute liver injury in livestock. This study evaluated whether Vitamin U could alleviate AFB1-induced hepatotoxicity in mice. AFB1 (3 mg/kg) reduced PLT, PCT, and EOS counts, caused hepatic vascular congestion, and decreased GSH-Px activity. Vitamin U (50 mg/kg) significantly improved these hematological parameters, alleviated central venous and sinusoidal congestion, increased T-SOD activity and upregulated IL-10 mRNA expression. However, Vitamin U did not significantly reverse AFB1-induced elevation of ALP or reduction in GSH-Px, nor did it affect ALT, AST, or protein levels of the Nrf2/Keap1 pathway. Transcriptomic analysis revealed enrichment of DEGs in immune- and cell cycle-related pathways, with no direct enrichment observed in the Nrf2/Keap1 pathway. Vitamin U upregulated Keap1 mRNA expression but did not alter KEAP1 protein levels. In conclusion, Vitamin U partially protects against AFB1-induced acute liver injury by ameliorating thrombocytopenia, vascular congestion, enhancing T-SOD activity, and upregulating IL-10 expression, providing preliminary experimental evidence for further investigation.

Veterinary Sciences doi: 10.3390/vetsci13070620

Authors: Tarek A. Ebeid Mohamed Tharwat Sohail Ahmad Ahmed O. Abbas Abdullah N. Alkhalaf Fahad A. Alshanbari

Sustainable rabbit production requires effective nutritional strategies to enhance productivity, health status, and immune competence. Following the restriction of antibiotic growth promoters, organic acids (OAs) have gotten increasing attention as promising functional feed additives due to their multiple biological roles. This review aims to offer a comprehensive overview of the functional roles of OAs in rabbit nutrition, with a focus on their effects on gut morphology, nutrient digestibility, intestinal microbiota, antioxidative status, immunity, and growth performance in growing rabbits. The OAs may modulate gut microbiota balance through inhibition of pathogenic bacteria and promotion of beneficial microbial populations, thereby contributing to the establishment of a balanced intestinal ecosystem. This effect is particularly important during the post-weaning period, a critical stage characterized by increased susceptibility to enteric disorders and associated economic losses. The OAs may also enhance digestive enzyme activities, leading to improving nutrient digestibility, feed efficiency, and reducing feed wastage. In addition, OAs have been shown to improve intestinal histomorphology through coordinated effects on epithelial proliferation, mucosal renewal, and tight junction integrity. Furthermore, OAs have been shown to modulate antioxidative status and immune responses, which are essential for maintaining intestinal health and overall production sustainability. Collectively, OAs represent a promising and viable nutritional strategy to enhance the sustainability and efficiency of rabbit production systems through their beneficial effects on gut health, nutrient utilization, immune competence, and antioxidative status.

Veterinary Sciences doi: 10.3390/vetsci13070619

Authors: Mª Carmen Aranda Paloma Jimena de Andrés Sergio Villanueva-Saz Mª de los Ángeles Jiménez

The European mink is a critically endangered mustelid. Species-specific hematological and biochemical reference intervals are lacking, complicating accurate health assessments in conservation programs. This study aimed to determine these reference intervals for European mink, following international guidelines, with 110 blood samples collected under isoflurane anesthesia, outside the breeding season, from 75 captive individuals (37 females, 38 males). Results were broadly comparable to the American mink and domestic ferret. Several parameters varied significantly according to sex, age, and body condition. Males showed higher hemoglobin (p = 0.031), whereas females, higher platelet (p = 0.014) and leukocyte counts, cholesterol, urea, alkaline phosphatase (ALP) (p ≤ 0.002), and globulin concentrations (p = 0.005). Males also had higher creatinine and alanine aminotransferase (ALT) (p < 0.001). Juveniles had higher leukocyte counts (p = 0.047) and ALP and calcium concentrations (p ≤ 0.016), while adults showed higher globulin and creatinine values (p ≤ 0.014). Overweight individuals exhibited increased hemoglobin and hematocrit (p ≤ 0.018) and mild electrolyte differences (p ≤ 0.045). Separate intervals for sex and age were not warranted for most analytes. These results provided specific reference intervals useful for essential health monitoring and clinical decision-making in conservation breeding, translocation, and reintroduction programs of European mink.

Veterinary Sciences doi: 10.3390/vetsci13070618

Authors: Nerea Cambeiro-Camarero Silvia Fernández-Martín Antonio González-Cantalapiedra

Preoperative anxiety may compromise perioperative welfare and anesthetic management in dogs. This randomized, blind clinical study evaluated the effects of oral tasipimidine on anxiety-related behaviors, handling quality, cardiovascular variables, anesthetic requirements and recovery in dogs undergoing elective ovariectomy. Thirty ASA I dogs were randomly allocated into three groups (n = 10). Treatment groups received tasipimidine (30 µg/kg PO) 45 min before admission, followed by morphine (0.3 mg/kg IM) combined with dexmedetomidine at either 2.5 µg/kg (GTa) or 5 µg/kg IM (GTb). Control dogs (GC) received an oral placebo followed by dexmedetomidine (5 µg/kg IM) and morphine (0.3 mg/kg IM). Anxiety was assessed using the Clinic Dog Stress Scale (CDSS), Visual Analog Scale (VAS), and Reactivity Evaluation Form (REF). Handling quality was assessed using a Numerical Rating Scale (NRS), and recovery quality using the Simple Descriptive Scale (SDS). Tasipimidine significantly reduced anxiety-related behaviors compared with controls, with lower CDSS (5 [0–10] vs. 13 [3–19]), VAS (4 [2–7] vs. 6.5 [2–9]), and REF scores (1 [1–3] vs. 2 [1–4]) (p < 0.05). Handling quality improved significantly in treated dogs with lower NRS (1 [1–2] vs. 2 [1–4]; p = 0.003). Preoperative heart rate was lower in tasipimidine-treated dogs (85 ± 19 bpm) than in controls (127 ± 23 bpm; p < 0.001). Recovery agitation requiring dexmedetomidine rescue sedation occurred in 30% of treated dogs versus 60% of controls. Propofol and fentanyl requirements did not differ significantly among groups (p > 0.05). Tasipimidine improved anxiety-related behavioral scores and handling quality in dogs; however, its cardiovascular effects and perioperative role warrant further investigation.

Veterinary Sciences doi: 10.3390/vetsci13070617

Authors: Haifeng Sun Qingqing Liu Shuyan Zhai Biyue Wu Zicheng Ma Yangyang Sun Kaiyuan Ye Haoyuan Wang Yanni Gao Xianwei Wang Juan Bai Ping Jiang

Porcine circovirus type 2 (PCV2) is the primary causative agent of a spectrum of porcine circovirus-associated diseases (PCVDs) and remains a major threat to the global swine industry. In this study, ten monoclonal antibodies (mAbs) targeting the Cap protein of PCV2 were generated and characterized. One mAb, designated 4C4, which exhibited high reactivity, strong neutralizing activity, and superior blocking efficacy, was selected for horseradish peroxidase (HRP) labeling. After optimizing the reaction parameters, a blocking ELISA was developed for the detection of the anti-PCV2 antibody. Using receiver operating characteristic (ROC) curve analysis, a cutoff value of 40% was established to distinguish positive from negative serum samples. The sensitivity and specificity of this blocking ELISA method were 98.66% and 100%, respectively. No cross-reactivity was observed with serum antibodies against classical swine fever virus (CSFV), porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), porcine reproductive and respiratory syndrome virus (PRRSV), or pseudorabies virus (PRV). Intra-assay and inter-assay repeatability tests yielded coefficients of variation (CVs) all below 10%, confirming the assay’s excellent reproducibility. Simultaneous testing of 312 clinical porcine serum samples using the developed bELISA and a commercial indirect ELISA kit revealed an overall coincidence rate of 99.04%. In addition, the percentage inhibition (PI) in the bELISA was strongly correlated with serum anti-PCV2 neutralizing antibody titers. In conclusion, the blocking ELISA developed herein demonstrates high sensitivity, strong specificity, and good reproducibility, serving as a potentially effective tool for the detection of the anti-PCV2 antibody and epidemiological investigation.

Veterinary Sciences doi: 10.3390/vetsci13070616

Authors: Wei Han Zhao Si Yeong Choi In Ho Kim

This study evaluated the effects of reducing dietary crude protein (CP) levels and protease supplementation on growth performance, nutrient digestibility, and fecal score in weaned piglets. A total of 200 crossbred weaned piglets (Duroc × [Landrace × Yorkshire]), with an initial body weight (BW) of 6.01 ± 1.14 kg, were used in a 31-day feeding trial. Piglets were assigned to four dietary treatments in a randomized complete block design with 10 replicates per treatment and five piglets per pen. The treatments were as follows: CON, basal diet; TRT1, low-protein diet with CP reduced by 1%; TRT2, low-protein diet with CP reduced by 2%; and TRT3, TRT2 supplemented with 0.1 g/kg protease. Piglets fed TRT2 had lower BW on days 7, 19, and 31 (p < 0.05) and lower average daily gain (ADG) during each growth phase and the overall period compared with CON (p < 0.05). Protease supplementation partially restored BW and ADG. However, average daily feed intake (ADFI), feed conversion ratio (FCR), apparent digestibility of dry matter (DM), nitrogen (N), and energy (E), and fecal score were not affected by dietary treatments (p > 0.05). In conclusion, reducing dietary CP by 2% impaired growth performance in weaned piglets, whereas protease supplementation partially alleviated this negative effect without significantly altering nutrient digestibility or fecal score.