Veterinary Sciences doi: 10.3390/vetsci13080833

Authors: Yongchang Li Xianyue Fu Zihan Wang Iqra Zafar Zeyun Cui Shiyun Li Xiujuan Feng Sen Yang Ercha Hu Chahan Bayin

From September 2024 to September 2025, 310 blood samples from cattle (n = 151), camels (n = 44), horses (n = 71), and wild mice (n = 44) across six Xinjiang counties/cities were screened by microscopy, genus-specific PCR, sequencing, and phylogenetic analysis. Overall, 46.77% (145/310) of the samples were positive for haemoprotozoan infection. Trypanosoma spp. showed the highest prevalence (25.48%, 79/310), followed by Theileria (16.13%, 50/310) and Babesia (5.16%, 16/310). Notable host-specific patterns emerged: camels had the highest T. evansi occurrence (86.36%), while wild mice had the highest Theileria prevalence (75%). Dual infections (mostly Trypanosoma + Theileria) occurred in 8.39% (26/310) of animals, and a triple infection was found in 0.32% (1/310). Phylogenetic analysis identified T. evansi as predominant trypanosome, clustering with isolates from India and Saudi Arabia; the major piroplasms identified were T. annulata, T. equi, and an unidentified Babesia sp. This study represents the first multi-host molecular survey in Xinjiang, revealing a high infection burden, diverse parasite species, and frequent co-infections.

Veterinary Sciences doi: 10.3390/vetsci13080832

Authors: Roberto Bava Stefano Ruga Giovanna Liguori Antonio Giordano Giancarlo Statti Mariangela Marrelli Vincenzo Musella Ernesto Palma Domenico Britti Carmine Lupia Fabio Castagna

Punica granatum L. (pomegranate) is one of the oldest medicinal plants known to humankind, valued across ancient civilisations for treating parasitic, microbial, and metabolic diseases. Its exceptionally rich phytochemical composition, dominated by punicalagins, ellagic acid, anthocyanins, flavonoids, piperidine alkaloids, and the unique conjugated fatty acid punicic acid, confers a remarkably broad spectrum of biological activities of direct relevance to contemporary veterinary medicine. While human-health applications have been extensively reviewed, a comprehensive synthesis of veterinary evidence across multiple species remains lacking. This review consolidates current preclinical and field knowledge on the pharmacological effects of pomegranate preparations in poultry, ruminants, swine, fish, companion animals, and laboratory models. In poultry—the most extensively studied taxon—dietary inclusion of pomegranate peel powder or extract consistently enhances growth performance, antioxidant status, and humoral immunity while exerting meaningful anticoccidial activity. The antiparasitic properties are compellingly supported by evidence against gastrointestinal nematodes of ruminants, tapeworms, schistosomes, and protozoa including Giardia, Cryptosporidium, and Leishmania spp., as well as monogenean fish parasites. Broad-spectrum antimicrobial activity extends to major veterinary pathogens such as Salmonella, Escherichia coli, Staphylococcus aureus (including MRSA), and Clostridium perfringens. Rodent models have validated antidiabetic, hepatoprotective, nephroprotective, and reproductive benefits, including improved post-thaw sperm quality and enhanced litter size. The safety profile is generally favourable at conventional doses, although high dietary inclusion elicits anti-nutritional effects from condensed tannins, and potential drug interactions via cytochrome P450 inhibition warrant clinical caution. Despite this substantial evidence, significant translational barriers persist, including extract heterogeneity, absence of pharmacokinetic data in target species, and scarcity of controlled clinical trials. By providing a species- and pathology-driven synthesis, this review identifies critical research priorities and highlights the immense potential of this ancient, accessible, and economically viable phytobiotic as a natural alternative to antibiotic growth promoters and synthetic antiparasitics in veterinary practice.

Veterinary Sciences doi: 10.3390/vetsci13080831

Authors: Zhe Jia Yanfei He Haijun Xu Cai Zhang Shunan Cuan

Global poultry consumption continues to rise, while worldwide bans on in-feed antibiotic growth promoters raise an urgent requirement for eco-friendly alternatives to guarantee production efficiency and food safety. Probiotics, live beneficial microorganisms that improve host intestinal health, are reviewed. We elaborate four core functional pathways of probiotics: competitive exclusion of pathogens, enhancement of intestinal barrier integrity, immune modulation and regulation of microbial metabolites such as short-chain fatty acids. Their mitigating effects against heat stress, suboptimal rearing environments, mycotoxin contamination, heavy metal exposure and immune stress are analyzed. We further evaluate the capacity of single and compound probiotics to control major poultry diseases. Early-life intervention strategies and innovative preparations (multistrain probiotics, synbiotics, postbiotics) are systematically summarized. Critical bottlenecks restricting industrial translation are highlighted, including empirical strain combination, non-standardized administration protocols, divergent evaluation indicators and single-factor laboratory challenge models inconsistent with actual farm conditions. Finally, we propose future research directions covering multi-omics-assisted strain screening, optimized delivery technology, unified industrial quality control standards and field verification under compound stress. This review offers integrated references for mechanistic research, strain development and precise industrial application of probiotics in sustainable antibiotic-free poultry breeding.

Veterinary Sciences doi: 10.3390/vetsci13080830

Authors: Sheng Yang Yikun Zhu Xin Wang Yufei Huang

Cellular inhibitor of apoptosis protein 1 (cIAP1) plays a crucial role in the apoptotic process. This study involved gene cloning, protein expression, bioinformatics analysis, and the preparation of a specific polyclonal antibody. First, the coding sequence of the duck cIAP1 was amplified by PCR and sequenced. Bioinformatics prediction indicated that this protein is a stable hydrophilic protein with no signal peptides and transmembrane domains. It contains characteristic BIR (bacterial IAP repeat) domains. The secondary structure consists of 43% random coils, 28% α-helix, and 8% β-strand, and its 3D structure is highly conserved compared with human and mouse cIAP1. Subsequently, the cIAP1 was cloned into the pET-28a vector. The recombinant His-tagged cIAP1 protein was successfully expressed in Escherichia coli BL21(DE3) upon induction and subsequently purified. Immunization of BALB/c mice with the purified protein emulsified in Freund’s adjuvant generated polyclonal antibodies. Indirect ELISA confirmed that the antibody titer exceeded 1:2,048,000. Additionally, Western blotting and immunofluorescence staining demonstrated that the antibody specifically recognized native duck cIAP1 protein without non-specific reactions with negative serum. This study successfully achieved prokaryotic expression of duck cIAP1 and produced a high-titer, highly specific mouse polyclonal antibody. This antibody is suitable for experiments such as Western blotting and immunofluorescence, providing important biological materials and a theoretical basis for further investigation of the anti-apoptotic mechanism of cIAP1 in major diseases of ducks.

Veterinary Sciences doi: 10.3390/vetsci13080827

Authors: Zhaoruiyi Li Xuan Zhou Yuxin Qin Lidan Wang Xinhui Zhang Sara Guesmia Yue Xie

Ascaridia galli is a major gastrointestinal nematode of poultry that causes significant economic losses worldwide. In this study, we characterized a new isolate, A. galli AgSC_2025, obtained from Xichuan black-bone chickens in China, using an integrative approach by combining morphology, scanning electron microscopy (SEM) and mitogenomics and then its complete developmental changes in eggs—the primary source of infection—was also documented using a high-resolution auto-microscopic imaging. The adult worms exhibited diagnostic characteristics of A. galli, including three distinct lips and a specific arrangement of male caudal papillae. Molecular analysis showed a 99.84% mitogenome identity with previously reported A. galli and positioned the isolate and A. galli in one branch which was sister to other congeneric species within the Ascaridia clade. Furthermore, the high-resolution auto-microscopic imaging system yielded a detailed and complete in vitro egg developmental dynamics with thirteen distinct stages, ranging from the initial 1-cell stage to the infective third-stage larva (L3); and further developmental timeline emphasized that the infective L3 larvae predominated by day 10, with 100% reaching this stage on day 13. Viability assays demonstrated a 79.8% ± 0.46% larval hatching rate and the high vitality of the liberated larvae among these infective eggs. These findings provide a comprehensive morphological and molecular reference for this A. galli isolate and also establish precise developmental biological parameters for egg embryonation, which should offer valuable insights for species-specific identification and the optimization of sanitation protocols to control avian ascaridiasis.

Veterinary Sciences doi: 10.3390/vetsci13080829

Authors: Akvilė Girdauskaitė Samanta Grigė Inga Sabeckienė Karina Džermeikaitė Justina Krištolaitytė Sigitas Japertas Rokas Macijauskas Lina Anskienė Bernadeta Škėlaitė Ramūnas Antanaitis

The transition period is a physiologically demanding stage in primiparous dairy cows, as the onset of lactation occurs simultaneously with continued growth. This longitudinal study initially enrolled 22 primiparous Holstein dairy cows. Eight cows that developed transition disorders during the monitoring period were excluded according to the predefined study protocol. Therefore, the final analysis included 14 clinically healthy primiparous Holstein dairy cows monitored from four weeks before calving to five weeks postpartum to characterize normal physiological adaptation during the transition period. Cows were monitored using continuous intraruminal sensors and automated milk analysis systems, while blood samples were collected weekly and analysed using linear mixed models with repeated measurements. Milk yield increased from 17.9 kg/day in week +1 to 26.3 kg/day in week +5 (a relative increase of 47%). Significant temporal changes were observed in iron (Fe), phosphorus (P), glucose (GLUC), triglycerides (TRIG), urea (UREA), albumin (ALB), and alanine aminotransferase (ALT). Serum Fe decreased from 30.9 µmol/L at week −2 to 14.5 µmol/L at week +2 (−53%), while P declined from 2.95 mmol/L at week +1 to 1.41 mmol/L at week +2 (−52%). Sensor data showed reduced physical activity (−59%) and increased water intake from 60.6 L/day in week −4 to 99.4 L/day in week +1, whereas rumination time and rumen pH remained relatively stable. Correlation analysis identified significant associations among milk yield, metabolic variables, and behavioural indicators. These findings suggest coordinated physiological adaptation during the transition period and support the use of combined blood and sensor-based monitoring to better understand early-lactation dynamics in primiparous dairy cows.

Veterinary Sciences doi: 10.3390/vetsci13080828

Authors: Valentina Gazzano Virginia Bellini Francesca Cecchi Maria Claudia Curadi Paolo Baragli Angelo Gazzano Asahi Ogi

Judgment bias tasks are widely used to assess affective state in adult dogs but have rarely been applied before adoption age. To test whether a spatial version is viable, 82 puppies from 17 litters were tested at 60 days of age; 80 provided valid observations for analysis. After discrimination training, positive (P), negative (N) and ambiguous (A) locations were presented in up to three trials; latency to the bowl was recorded (30 s cut-off). Latencies from the first two trials (476 observations, 85 right-censored) were analyzed with mixed-effects Cox proportional hazards models including puppy and litter as random intercepts. Median recorded times were 2.54 s (P), 13.25 s (N) and 3.86 s (A), with timeouts in 0%, 38.4% and 15.2%, respectively. Cue location affected approach rate (χ2 = 207.03, df = 2, p < 0.001); hazard ratios were N versus P 0.11 (95% CI 0.09–0.15), A versus P 0.32 (0.25–0.41) and N versus A 0.36 (0.27–0.47), all significant (Holm-adjusted). Trial number (p = 0.076) and sex (p = 0.483) were not significant. Random-effect variances were 0.180 (puppy) and 0.261 (litter). Puppies discriminated between outcome-associated locations and approached the ambiguous location at an intermediate rate, indicating that the task is feasible at 60 days of age.

Veterinary Sciences doi: 10.3390/vetsci13080826

Authors: Zhaolong Li Hongyang Zhao Peimin Li Song Peng Junru Mo Guiheng Mei Meiqin Li Fengqiang Lin Quanwang Wu Dexicuomu Danzengzhuoga Sangjiebazhu Pubuzhaxi

The ban on antibiotic growth promoters has created an urgent need for effective alternatives to maintain intestinal health and production performance. This exploratory study evaluated the effects of a novel antibiotic alternative additive at graded inclusion levels (control, 1%, 3%, 5%) on growth performance, intestinal histomorphology, gut microbiome ecology, and host metabolome in Tibetan pigs (n = 40). Forty Tibetan pigs were randomly assigned to four dietary treatment groups for a 21-day feeding trial. Growth performance, intestinal histopathology, metatranscriptomic sequencing (bacteria, viruses, fungi), and untargeted metabolomics were analyzed. Tibetan pigs fed the 3% additive diet exhibited the highest numerical average daily gain (0.197 vs. 0.148 kg/d, +33.1%) and lowest feed conversion ratio (2.61 vs. 3.59, −27.3%), with overall treatment effects reaching statistical significance (ADG: p = 0.027; FCR: p = 0.023). Metatranscriptomic profiling revealed fundamentally distinct microbial communities between the duodenum and colon. Multi-omics integration identified 79 significant microbiome-pathology associations in the duodenum and 38 in the colon, with SCFA-producing bacteria positively correlated with mucosal architectural parameters. The multi-omics network revealed a core-periphery architecture with Eubacterium, Lactobacillus, and Escherichia as keystone hubs. Dietary supplementation at 3% showed the most favorable effects on growth performance and metabolic modulation without compromising intestinal structural integrity, supporting its potential as a viable antibiotic alternative.

Veterinary Sciences doi: 10.3390/vetsci13080825

Authors: Haoyu Wang Yong Zhou Peng Chen Liping Zhang Wen Zhu Mingyang Xue Yan Meng Zhenyu Huang Chen Xu Yuding Fan Chao Pei Nan Jiang

The high mortality associated with largemouth bass ranavirus (LMBRaV) disease requires rapid and sensitive diagnosis methods to prevent virus spread and subsequent outbreak. The recombinase polymerase amplification combined with the CRISPR/Cas12a system (RPA-CRISPR/Cas12a) assay and RPA-CRISPR/Cas12a, coupled with lateral flow dipstick (RPA-CRISPR/Cas12a-LFD) assay for LMBRaV, were established in this study for low viral load surveillance. The major capsid protein (mcp) gene is highly conserved and is widely used as the target sequence in LMBRaV detection. First, based on the mcp gene sequence of LMBRaV, three candidate crRNAs were designed. Among them, the crRNA-2, which exhibited the highest cleavage activity, was selected through dual evaluation of fluorescence signal intensity and LFD color development. Then, the key reaction conditions for both assays were optimized as follows: LbCas12a protein concentration of 150 nM, crRNA-2 concentration of 200 nM, and ssDNA reporters’ concentration of 200 nM. Moreover, specificity evaluation showed that both combined assays specifically recognized LMBRaV, with no cross-reactivity detected against other aquatic pathogens, such as CyHV-2, GSIV, ISKNV, WSSV, GCRV II or CrERV. The detection limit was 1 copy/μL of DNA sample for both RPA-CRISPR/Cas12a and RPA-CRISPR/Cas12a-LFD assays. Finally, validation using 24 clinical samples (16 positive, eight negative) showed that both LMBRaV RPA-CRISPR/Cas12a assays achieved 100% detection rate, whereas conventional PCR detected 13 positive samples (detection rate 81.26%). ddPCR served as the reference method for clinical validation. Therefore, the LMBRaV RPA-CRISPR/Cas12a assay and the RPA-CRISPR/Cas12a-LFD assay provide sensitive, specific, and easy-to-operate methods for the rapid detection of LMBRaV.

Veterinary Sciences doi: 10.3390/vetsci13080824

Authors: Jean-François Da-Costa Arnaud Lebret Gwenaël Boulbria Valérie Normand Justine Favrel Mathieu Brissonnier Nadia Amenna-Bernard Sophie Labrut Marion Mosca Didier Pin

This case report describes the investigation of two cases of cutaneous vasculitis in sows following vaccination. The herd was porcine reproductive and respiratory syndrome virus (PRRSV)-stable, with routine PRRSV-1 and porcine circovirus type 2 (PCV2) sows’ vaccination. Two sows developed red-to-purple macules on the udder, flanks, and hind limbs, occurring two and three days after vaccination, respectively. Diagnostic investigations included biochemistry, complete blood count, PCR, immunohistochemistry (IHC), serology, histopathology, Gram staining, and immunofluorescence (IF) to assess the involvement of type III hypersensitivity. Clinical examination and herd evaluation revealed no significant abnormalities. Both sows showed biochemistry results within the reference intervals. One sow had lymphocytosis. All PCR PCV2, PCV3 and PRRSV-1/2 on serum and IHC PCV2 conducted on skin biopsies were negative, although both animals were seropositive for PCV2. Histopathology revealed necrotizing leukocytoclastic vasculitis, and Gram staining detected no bacteria. IF failed to identify immune complex or complement deposition. However, type III hypersensitivity could not be excluded, as immune complexes are transient and biopsies were collected about 48 h after lesion onset. Overall, the findings support a vaccine-related adverse reaction. Vaccine adverse effects should therefore be considered in the differential diagnosis of cutaneous vasculitis in swine.

Veterinary Sciences doi: 10.3390/vetsci13080823

Authors: Yuhong Wu Yuhui Ma Yiyi Zan Xu Bai Jing Li Hai Li

The equine gastrointestinal microbiome plays an essential role in digestion, energy metabolism, immune regulation, and maintenance of intestinal homeostasis. Increasing evidence suggests that microbial-derived metabolites provide a functional link between diet, the microbiome, and host physiology. This narrative review summarizes current knowledge on how dietary factors, including structural carbohydrates, non-structural carbohydrates, protein, and lipids, influence microbial function and metabolite production in horses. Relevant publications available up to June 2026 were identified through searches of PubMed, Web of Science, Scopus, and Google Scholar and were narratively synthesized according to dietary factors, microbial metabolites, associated diseases, and nutritional interventions. Emphasis is placed on major microbial metabolites, including short-chain fatty acids, endotoxins, bile acid derivatives, tryptophan metabolites, and nitrogenous fermentation products, and their potential roles in health and disease. Current evidence supports a central role for the nutrition–microbiota–metabolite axis in gastrointestinal disorders such as colic, colitis, equine gastric ulcer syndrome, and carbohydrate-associated laminitis, whereas its involvement in equine metabolic syndrome and the gut–lung axis remains less well defined. The review also discusses microbiome-targeted nutritional interventions and emerging multi-omics approaches that are improving our understanding of microbial function. Collectively, these findings highlight the potential of microbiome-informed nutritional strategies to support equine health, welfare, disease prevention, and athletic performance.

Veterinary Sciences doi: 10.3390/vetsci13080822

Authors: Hyeong-Wook Moon Chi-Youn Song Jong-Myung Lee Kwang-Rae Jo Geun-Bo Park Hye-Soo Shim Hwi-Yool Kim Jung-Moon Kim

Primary hepatic neoplasms in dogs, while uncommon, are occasionally associated with severe hypoglycemia, although the underlying mechanism may be difficult to establish. A 7-year-old castrated male Maltese dog weighing 3.6 kg was referred for recurrent bilateral pelvic limb weakness, episodic collapse, and abdominal distension. Severe hypoglycemia was documented, with a blood glucose concentration of 26 mg/dL. Computed tomography revealed a large mass arising from the right lateral liver lobe, with no identifiable pancreatic mass or distant metastasis. The serum insulin concentration measured during hypoglycemia was low at 1.0 μU/mL, making insulinoma less likely. Glycemic control temporarily or partially improved during prednisolone treatment; however, severe hypoglycemia recurred after prednisolone discontinuation before surgery. The hepatic mass was surgically excised. Dextrose-containing intravenous fluids were maintained for 6 h postoperatively, and no recurrent clinically significant hypoglycemia was documented after their discontinuation. Histopathologic examination revealed a poorly differentiated neoplasm with mesenchymal morphology, occasional cytoplasmic vacuolation, and no unequivocal lipoblasts. The neoplastic cells showed diffuse vimentin immunoreactivity, weak cytoplasmic S-100 immunoreactivity in a subset of cells, and no immunoreactivity for smooth-muscle actin or hepatocyte paraffin 1. These findings raised the possibility of adipocytic differentiation but were insufficient to establish a specific tumor lineage; therefore, the lesion was classified as a primary hepatic neoplasm of uncertain histogenesis. At 247 days after surgery, the dog remained normoglycemic without supplemental glucose or glucocorticoid treatment, and no imaging evidence of local recurrence or distant metastasis was identified. The postoperative course supported an association between the hepatic neoplasm and hypoinsulinemic hypoglycemia. Non-islet-cell tumor hypoglycemia was suspected but remained unconfirmed because an insulin-like growth factor-mediated mechanism was not demonstrated.

Veterinary Sciences doi: 10.3390/vetsci13080821

Authors: Mudathir Y. Abdulrahman Nasir A. Ibrahim Mohamed Osman Abdalrahem Essa Saber Y. Adam Raza Mohai Ud Din Abdelkareem A. Ahmed Rifat Ullah Jan Hamdi Bendif Nosiba S. Basher Ahmed A. Saleh Hosameldeen Mohamed Husien Mengzhi Wang

Moringa oleifera leaf extract (MOLE), including polysaccharides, polyphenols, amino acids and other extracts, are increasingly being used as feed additives in ruminant nutrition due to their high profiles of bioactive compounds. This comprehensive narrative review aims to review current research on their use in ruminant production regarding their productive performance, immune status, rumen microbiota and fermentation. Dietary supplementation with MOLE enhances growth rates and feed conversion efficiency, as well as immune status, milk yield and rumen microbiology, in ruminants by improving nutrient digestibility and metabolic efficiency. Additionally, MOLE exerts significant immunomodulatory effects, and studies indicate that immunoglobulins, antioxidants, cytokines, and enzymes reduce oxidative stress markers and incidence of subclinical diseases in dairy animals. Furthermore, supplementation with MOLE often increases milk fat and protein content while reducing somatic cell counts (SCCs) in milk. MOLE enhance the function of the rumen fermentation profile by increasing volatile fatty acid (VFA) production, especially propionate, and also regulating the stability of rumen pH. Additionally, it also selectively promotes microbial community diversity, enhances the population of cellulolytic bacteria and has a suppression function in protozoa and methanogens. Moreover, MOLE also contributes to improving fiber degradation and reducing the emissions of methane. In conclusion, MOLE is considered as a viable natural strategy to promote productivity, immune function, health, and environmental sustainability while decreasing methane production in ruminant production.

Veterinary Sciences doi: 10.3390/vetsci13080820

Authors: Shuairan Zhang Yu Zhang Cai Yin Xiaona Zang Minjing Jin Yawen Wu Taotao Bai Long Ma Yong Shi Zhixin Li Xiaoliang Wang

To establish a rapid nucleic acid detection method for field screening of Brucella spp., a visual assay based on recombinase polymerase amplification (RPA) combined with CRISPR/Cas12a was developed. Four pairs of RPA primers and two crRNAs were designed according to the conserved Brucella bcsp31 sequence. The optimal RPA primer–crRNA combination was screened using agarose gel electrophoresis and fluorescence readout, and the concentrations of Cas12a protein, crRNA, and ssDNA fluorescent reporter were optimized. RPA primer pair 2 showed the best amplification performance, and RPA2-crRNA1 was selected as the optimal combination for the assay. The final reaction conditions were 200 nM Cas12a protein, 200 nM crRNA, and 250 nM ssDNA fluorescent reporter. The limit of detection of the established method was 2.47 × 100 copies/μL. No cross-reactivity was observed with Mycoplasma capricolum subsp. capripneumoniae, Chlamydia abortus, Chlamydia psittaci, or Salmonella enterica serovar Typhimurium. Among 56 clinical sheep nasal swab samples collected during routine surveillance, qPCR identified 22 positive and 34 negative samples, while the RPA-CRISPR/Cas12a assay identified 20 positive and 36 negative samples. The positive agreement, negative agreement, overall agreement, positive predictive value, negative predictive value, and Cohen’s kappa value were 90.91%, 100.00%, 96.43%, 100.00%, 94.44%, and 0.924, respectively. These findings provide preliminary evidence that the developed bcsp31-targeted RPA-CRISPR/Cas12a assay is rapid and visually interpretable and may be useful for preliminary Brucella spp. screening in routine veterinary surveillance settings. Larger-scale validation using different sample types and samples from different regions is still required.

Veterinary Sciences doi: 10.3390/vetsci13080818

Authors: Tariq Sohail Mohamed Tharwat Aftab Shaukat Nourhan Nassar Fuhao Chen Xiaomei Sun Fahad A. Alshanbari Yongjun Li

The successful use of Assisted Reproductive Technologies (ARTs) such as artificial insemination (AI) in small ruminants and other mammals depends on many factors, the most important of which is the quality of the semen used. The liquid storage and cryopreservation of spermatozoa at lower temperatures are associated with artificial insemination and rapid genetic improvement programs in the sheep production industry. Several studies have reported increased lipid peroxidation (LPO) and highly reactive oxygen species (ROS) production during liquid storage or cryopreservation of ram semen, leading to oxidative stress (OS), decreased antioxidant defense, and changes in sperm quality parameters like biokinetic and biochemical characteristics, viability, functional membrane and DNA integrity, along with mitochondrial activity. Therefore, supplementing ram semen extenders with exogenous antioxidants before preservation could mitigate this harmful effect. Various in vitro studies have reported improvements in ram sperm quality parameters like motility indexes, vitality, functional membrane/DNA integrity, antioxidant enzyme activity, total antioxidant content, mitochondrial activity, in vivo/in vitro fertility with significant decline in sperm abnormality, free radical production, LPO, ROS, apoptosis rate, and cytochrome C release from the mitochondrial matrix after the addition of various natural and synthetic antioxidant (vitamins, glutathione, taurine, pyruvate, melatonin, cysteine, selenium, zinc, plant extracts, sugars, amino acids, polyphenols) substances during preservation. Therefore, this review summarizes recent findings on oxidative stress-induced damage to sperm quality parameters in various ram breeds during chilling storage and cryopreservation. Moreover, supplementing basic semen extenders with different non-enzymatic antioxidant substances as a method to maintain sperm quality—along with their efficacy in reducing or preventing sperm damage during preservation—was discussed in detail.

Veterinary Sciences doi: 10.3390/vetsci13080819

Authors: Nestor Acosta Teresa Guarda

National evidence on cattle vaccination, disease-control practices, and producer-reported disease remains limited in Ecuador. This study assessed holding-level participation in preventive cattle-health activities and factors associated with self-reported disease, focusing on Manabí. We analysed anonymised public-use microdata from the 2025 Continuous Agricultural Production and Area Survey (ESPAC); all four outcomes were based on producer reports and were not independently verified against veterinary or administrative records. The principal design-adjusted analysis included 10,588 positive-weight holdings across continental Ecuador. Survey-weighted proportions and multivariable logistic regression models incorporated expansion weights, strata, and primary sampling units. Estimated participation in the official foot-and-mouth disease vaccination campaign, other cattle vaccination, and disease-control activities was 86.38%, 55.96%, and 64.74%, respectively; 11.22% of holdings reported a principal cattle disease. Larger herd size was positively associated with all four outcomes (all p < 0.001). Compared with the Coast, holdings in the Amazon and Highlands had lower adjusted odds of other vaccination and disease-control activities and higher odds of self-reported disease (all p ≤ 0.002). Manabí holdings had higher adjusted odds of all three preventive outcomes than holdings in the other Coast provinces (all p < 0.001), but not of self-reported disease (p = 0.493). Preventive engagement was strongest for the centrally organised foot-and-mouth disease campaign. These findings identify territorial and farm-level inequalities relevant to veterinary-service delivery and surveillance, without establishing animal-level coverage, confirmed prevalence, or causal effects.

Veterinary Sciences doi: 10.3390/vetsci13080817

Authors: Kaixin Zhang Chang Wang Richard Laven Baiyu Li Meng Li D. Aaron Yang

Population-level information on feline morbidity in China remains limited despite the rapid growth of the pet cat population. This study described morbidity patterns among insured cats in China using insurance claims from a major pet insurance provider, with diagnoses derived from veterinary records. Disease events were classified into hierarchical diagnostic categories and subcategories using disease names and clinical descriptions. The study population included 163,890 insured cats contributing 194,284 cat-years at risk. A total of 42,867 disease events were identified in 31,870 cats, and ~ 19.5% of insured cats had at least one disease event during follow-up. The overall incidence rate was ~22 events per 100 cat-years. Digestive disorders were the most commonly claimed disorders, followed by urinary disorders, infectious diseases, respiratory disorders, and dermatological disorders. Gastroenteritis and lower urinary tract disease were the most frequent diagnostic subcategories. Cats aged less than one year had the highest overall incidence rate. Male cats had a higher incidence rate than female cats, mainly reflecting the higher burden of lower urinary tract disorders. Descriptive differences were also observed among British Shorthair, Maine Coon, and Devon Rex cats. This study provides the first broad insurance-based description of feline morbidity across multiple diagnostic categories in China. The findings offer baseline evidence for future disease-specific epidemiological studies.

Veterinary Sciences doi: 10.3390/vetsci13080816

Authors: Xiujun Duan Rui Zhu Youqing Bian Xiaoming Li Guobo Sun Yue Wang Qingyi Zhou Lei Zhang

As a high-value meat duck breed, Muscovy ducks exhibit marked individual differences in egg production that limit the economic efficiency of commercial farming. To elucidate the underlying mechanisms, we integrated phenotypic observation, ovarian transcriptome sequencing, and cecal 16S rRNA gene amplicon sequencing to compare high- and low-laying black-feathered Muscovy duck populations. We recorded 40-day egg production, assessed ovarian morphology, quantified gene expression profiles, and characterized microbial community structure. High-laying ducks produced significantly more eggs (24.60 ± 9.22) than low-laying ducks (15.80 ± 6.64, p < 0.01), with larger ovaries and a greater number of follicles at all developmental stages. Transcriptomic analysis revealed 823 DEGs, with KEGG enrichment implicating pathways governing ovarian physiology, including MAPK, Calcium, Notch, and Wnt signaling. Microbial profiling demonstrated significant differences in α/β diversity between groups, with high-laying ducks exhibiting an elevated Firmicutes/Bacteroidota ratio and decreased abundance of Ligilactobacillus. Integrative correlation analysis identified significant associations between Ligilactobacillus abundance and the expression of Arhgap27 and HOXA10, two genes linked to reproductive function, and highlighted the glycolysis/gluconeogenesis pathway as a convergently enriched signaling cascade shared between gut microbiome and ovarian transcriptome. This multi-omics study uncovers the molecular and microbial basis of egg production variation in Muscovy ducks, offering potential targets for improving breeding efficiency through microbiota-directed or gene-based interventions. These findings deepen our understanding of host–microbe interactions in poultry reproduction and identify candidate genes and microbial taxa that may serve as potential targets for future nutritional or genetic interventions to enhance laying performance in commercial duck production.

Veterinary Sciences doi: 10.3390/vetsci13080815

Authors: Qiangqiang Wang Xiaoxuan Feng Yurun Su Naiwen Zhang Yevheniia Dudnyk Hongxuan He

Cancer is increasingly recognized as an emerging concern in wildlife health and biodiversity conservation in the Anthropocene. Although traditionally viewed as an individual disease associated primarily with aging, wildlife cancer is shaped by complex interactions among environmental changes, species-specific evolutionary adaptations, and ecological processes. This review synthesizes current knowledge on the ecological and evolutionary drivers of wildlife cancer by integrating evidence from comparative oncology, environmental toxicology, wildlife pathology, and conservation biology. We examine how anthropogenic stressors, including pollution, habitat degradation, climate change, and infectious agents, influence cancer susceptibility in wild populations, and summarize intrinsic mechanisms underlying interspecific variation in cancer vulnerability, including Peto’s paradox, enhanced tumor suppression, and adaptive immune surveillance. We further highlight major methodological challenges, including limited surveillance capacity, fragmented datasets, taxonomic biases, and insufficient integration between cancer biology and conservation science. Finally, we discuss emerging interdisciplinary approaches, such as standardized monitoring frameworks, multi-omics technologies, artificial intelligence-assisted diagnosis, and One Health-based strategies, to advance wildlife cancer research and management. Collectively, this review positions wildlife cancer as an important ecological and evolutionary phenomenon and provides perspectives for incorporating cancer surveillance into biodiversity conservation and ecosystem health assessment.

Veterinary Sciences doi: 10.3390/vetsci13080814

Authors: Jalil Ghassemi Nejad Farzad Ghanbari Si-Hyeong Sun

Hair-based biomarkers offer a non-invasive window into chronic stress physiology, yet the influence of sampling body site on hair cortisol, dehydroepiandrosterone (DHEA), and their molar ratio has not been systematically characterized in domestic dogs. This study compared these three biomarkers across three anatomical sites—shoulder (mid-scapula), neck (dorsal midline), and rump (sacroiliac region)—in 30 neutered Anatolian shelter dogs (2–3 years; 15 males, 15 females; 30 dogs total, with each dog sampled at all three body sites) housed in individual kennels over eight weeks. Hair samples were collected at week 0 and week 8, extracted by methanol pulverization, and quantified by validated ELISA protocols. Hair cortisol differed significantly by site (p < 0.001), as did DHEA (p < 0.01) and cortisol-to-DHEA ratios (p < 0.001). Cortisol decreased modestly over eight weeks (p < 0.05). DHEA was inversely correlated with cortisol (r = −0.62, p < 0.01). The shoulder site yielded the highest absolute concentrations and the greatest cortisol-to-DHEA ratio, combined with the lowest within-site coefficient of variation (CV = 11.2% vs. 13.8% at neck and 15.6% at rump), jointly supporting its designation as the preferred anatomical sampling location. The cortisol-to-DHEA ratio is identified as a nuanced welfare indicator that integrates both allostatic load and adaptive resilience capacity, with implications for standardized non-invasive stress assessment protocols in shelter medicine.

Veterinary Sciences doi: 10.3390/vetsci13080813

Authors: Shaukat Ali Shaukat Jaspal Hanan Al-Khalaifah Muhammad Mubashar Shaukat Robina Shaukat Tahmina Shaukat Shabana Naz Rifat Ullah Khan Vincenzo Tufarelli Caterina Losacco Luciana Rossi Mutassim M. Abdelrahman

This study investigated age- and sex-related variations in pituitary gonadotroph morphometry and serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) concentrations in 90 clinically healthy dromedary camels (Camelus dromedarius). Animals were grouped into three age categories (2–4, 5–10, and ≥11 years), with equal representation of males and females. The precise reproductive status of the female camels (e.g., estrous stage or pregnancy status) was not determined and should be considered when interpreting the endocrine findings. Immunocytochemical analysis revealed significant age- and sex-related differences in gonadotroph number and morphometric characteristics (p < 0.05). Gonadotroph counts were 8.3 ± 0.4, 9.4 ± 0.5, and 8.1 ± 0.4 cells in the 2–4, 5–10, and ≥11-year age groups, respectively, with males showing a higher overall count than females (9.1 ± 0.4 vs. 8.2 ± 0.3; p < 0.05). Overall cell size, cell area, cell volume, nucleus area, and nucleus volume were also greater in males than females (p < 0.05). The largest nucleus size, nucleus area, and nucleus volume were observed in camels ≥11 years of age, with values of 4.7 ± 0.1 µm, 18.5 ± 0.8 µm2, and 60.3 ± 4.1 µm3, respectively. Ultrastructural analysis further showed a significant sex difference in granule size (58.0 ± 0.7 vs. 52.2 ± 0.8 nm) and granule percentage (56.2 ± 3.6 vs. 43.5 ± 3.3% in males and females, respectively; p < 0.05). Serum FSH concentrations were higher in males than females (4.8 ± 0.1 vs. 4.2 ± 0.1 µIU/mL; p < 0.05), whereas LH concentrations were 1.2 ± 0.1 and 1.2 ± 0.1 µIU/mL in males and females, respectively. LH concentration was highest in 5–10-year-old males (1.3 ± 0.2 µIU/mL), while FSH concentration was highest in ≥11-year-old males (4.8 ± 0.2 µIU/mL). Pearson correlation analysis demonstrated significant positive associations between gonadotroph morphometric parameters and circulating gonadotropins, with correlations ranging from 0.52 to 0.74 for cell/nuclear measurements with LH and FSH. Overall, the findings demonstrate marked age- and sex-associated differences in gonadotroph morphology, ultrastructure, and circulating gonadotropin concentrations in dromedary camels, with males generally exhibiting greater gonadotroph dimensions and secretory granule characteristics. These structural and endocrine differences provide evidence of age- and sex-related variation in pituitary gonadotrophic activity, although the findings should be interpreted in light of the cross-sectional design and the undetermined reproductive status of female camels. Overall, these findings demonstrate pronounced age- and sex-associated differences in pituitary gonadotroph morphology, ultrastructure, and endocrine activity, with males generally exhibiting greater cellular and secretory characteristics. The findings provide novel morphological and endocrine evidence of reproductive aging and sexual dimorphism in the dromedary camel pituitary and may contribute to improved understanding of reproductive function across different life stages. However, interpretation should consider the cross-sectional design and the inability to determine the precise reproductive stage of female camels before slaughter.

Veterinary Sciences doi: 10.3390/vetsci13080812

Authors: Ömer Barış İnce Ahmet Sait Alparslan Ayaz

Pestivirus infections, including those caused by Border disease virus (BDV), are associated with reproductive disorders, neonatal losses, and persistent infections in sheep, potentially resulting in substantial economic losses. This study aimed to determine the animal- and herd-level pestivirus seroprevalence among sheep in Konya Province, Türkiye, to investigate evidence of current infection, and to identify factors associated with seropositivity. Serum and whole-blood samples were collected from 280 sheep in 16 herds. Serum samples were tested for pestivirus antibodies by competitive ELISA, while whole-blood samples were examined for pestiviral antigen and RNA by antigen ELISA and real-time RT-PCR, respectively. Epidemiological associations were evaluated using generalized estimating equations (GEE), with herd included as the clustering variable. Pestivirus antibodies were detected in 168 of 280 sheep, corresponding to an animal-level pestivirus seroprevalence of 60.00% (95% confidence interval (CI): 54.16–65.57). At the herd level, 14 of 16 herds (87.5%) had at least one seropositive animal, while herd-specific antibody seropositivity ranged from 0% to 82.76%. Four animals were antigen-reactive at the initial sampling, and one remained so at the 15-day retest; however, all samples were negative by real-time RT-PCR. Therefore, persistent infection could not be confirmed. In the multivariable GEE model, animal age, herd disease history, and the introduction of new animals into the herd were significantly associated with pestivirus seropositivity. Together with previous studies from Türkiye, our findings indicate that pestivirus exposure remains common in sheep populations and warrants continued regional surveillance. Serological screening should therefore be complemented by direct detection methods, including antigen ELISA and real-time RT-PCR, when investigating current or persistent pestivirus infection. Informing herd owners about pestivirus infections and appropriate biosecurity measures may also contribute to reducing their potential impact on animal health and production.

Veterinary Sciences doi: 10.3390/vetsci13080811

Authors: Pei Wen Haiyang Wu Feitao Dang Jinbo Sun Hongwei Shi Yi Yan Jiayin Lu Xiaomao Luo Yanjun Dong Haidong Wang

Irisin is a myokine derived from fibronectin type III domain-containing protein 5 (FNDC5) and is involved in energy metabolism, muscle development, and oxidative regulation. Although irisin has been extensively studied in mammals, its potential application in poultry production remains unclear. Enhancing growth performance and physiological resilience is critical for male white-feathered broiler chickens under intensive production systems. This study investigated the effects of dietary irisin supplementation on growth performance and physiological responses in male white-feathered broiler chickens. A total of 180 broilers were randomly allocated to three treatments, with six replicate pens of ten birds per treatment: basal diet and basal diet supplemented with 5 and 10 mg/kg irisin for 42 days. At 42 days, one bird from each replicate pen was sampled for carcass, meat-quality, serum, immune, and microbiota analyses. Dietary irisin supplementation significantly increased final body weight and average daily gain compared with the control group. Feed conversion ratio decreased from 1.51 in the control group to 1.46 in the 5 and 10 mg/kg groups. Carcass evaluation showed higher slaughter yield and breast muscle yield in irisin-supplemented birds, and cooking loss was significantly reduced in both breast and thigh muscles. Serum superoxide dismutase (SOD) activity was elevated in irisin-treated groups. In addition, immunoglobulin Y (IgY) and immunoglobulin M (IgM) concentrations were significantly increased. No significant treatment effects were observed for total antioxidant capacity (T-AOC), catalase (CAT), malondialdehyde (MDA), inflammatory cytokines (IL-6, TNF-α), or immune organ indices (thymus, spleen and bursa of Fabricius). Gut microbiota analysis showed numerical differences in the relative abundance of several major bacterial taxa among treatments. In conclusion, dietary irisin supplementation was associated with improved growth performance and selected carcass, meat-quality, antioxidant, immunoglobulin, and cecal microbiota outcomes in male white-feathered broiler chickens, supporting further investigation of its potential as a functional feed additive in poultry nutrition.

Veterinary Sciences doi: 10.3390/vetsci13080810

Authors: Rebecca Leandri Giorgia Rosato Teresa Chianese Evaristo Di Napoli Manuela Martano Karen Power

Feline mammary tumors are the third most common neoplasms in female cats and they are characterized by high aggressiveness and high metastatic rates. Given the pivotal role of iron in human breast cancer development, in this study, we preliminarily explore the immunohistochemical expression of proteins involved in iron uptake, storage and efflux [Transferrin Receptor 1 (TfR1), Transferrin Receptor 2 (TfR2), ferritin (FTH1), and ferroportin (SLC40A1)] in non-metastatic and metastatic feline mammary carcinomas and their tributary lymph node and the possible correlation with hypoxia-inducible factor-1 (HIF-1). Our results showed an increased expression of TfR1 and TfR2 in relation to tumor progression. FTH1 immunolabeling was mainly observed in cancer cells delimiting necrotic areas and in lymph node metastasis, indicating greater iron storage possibly associated with hypoxic environments, as suggested by increased expression of HIF-1α. Also, an increase in SLC40A1 labeling in tumoral cells suggested greater iron efflux. Although preliminary, our results underline interesting differences between feline normal and tumoral mammary tissues, which could pave the way to further in vitro studies to better understand the role of iron in the progression of feline mammary tumors.

Veterinary Sciences doi: 10.3390/vetsci13080809

Authors: Serpil Mişe Yonar Mehmet Nuri Çakmak Duygu Tanrıkulu Mücahit Eroğlu Muhammet Enis Yonar

Copper sulfate (CuSO4) is one of the most widely used chemotherapeutic agents in aquaculture and aquatic veterinary medicine because of its broad-spectrum antiparasitic, antifungal, and antimicrobial activities. However, repeated exposure may induce oxidative stress and impair hematological and immune functions, thereby compromising fish health. This study evaluated the effects of pre-exposure and post-exposure dietary ellagic acid (EA) supplementation on CuSO4-induced hematological, immunological, and oxidative stress alterations in common carp (Cyprinus carpio). A total of 150 fish were randomly allocated to five experimental groups: Control, EA, CuSO4, pre-exposure EA + CuSO4, and post-exposure CuSO4 + EA. Fish were fed a diet supplemented with 100 mg EA kg−1 feed for 14 days either before or after repeated CuSO4 exposure (1 mg L−1, 30 min day−1 for seven consecutive days). Hematological variables, innate immune responses, and oxidative stress biomarkers were subsequently evaluated. CuSO4 exposure significantly decreased red blood cell (RBC) count, hemoglobin (Hb), hematocrit (Ht), phagocytic activity (PA), phagocytic index (PI), total plasma protein (TP), immunoglobulin M (IgM), bactericidal activity (BA), lysozyme (LYZ), superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione S-transferase (GST), and reduced glutathione (GSH), while significantly increasing mean corpuscular hemoglobin concentration (MCHC), white blood cell (WBC) count, respiratory burst activity (NBT), myeloperoxidase (MPO), and malondialdehyde (MDA) (p < 0.05). Dietary EA supplementation markedly alleviated these alterations by restoring hematological homeostasis, improving innate immune responses, reducing lipid peroxidation, and reinforcing antioxidant defense systems in the liver, kidney, spleen, and gill. Both pre-exposure and post-exposure EA supplementation were associated with improvements in the evaluated biomarkers under the experimental conditions of the present study. However, because the two supplementation strategies were not evaluated against a time-matched CuSO4 recovery control group, the present study does not permit a direct comparison of their relative efficacy. These findings demonstrate that dietary EA was associated with improvements in hematological, immunological, and oxidative stress biomarkers following experimental CuSO4 exposure, potentially through antioxidant and immunomodulatory mechanisms. Therefore, dietary EA may serve as a promising functional feed additive to support antioxidant and immune status during experimental CuSO4 exposure in common carp. Further studies including pathogen challenge models are required before its use as an adjunct to CuSO4 treatments can be recommended.

Veterinary Sciences doi: 10.3390/vetsci13080808

Authors: Zhi-Jiang Dong Ying-Ying Ren Wen-Hua Qi

The FMD is an endangered species under first-class national protection in China. Comparative genomic investigation of microsatellite (SSR) in CDS may provide insights into adaptive evolutionary mechanisms and may inform conservation management strategies for captive populations. Here, we analyzed the FMD genome alongside five closely related ruminants: cattle (Bos taurus), red deer (Cervus elaphus), white-tailed deer (Odocoileus virginianus), sheep (Ovis aries), and goat (Capra hircus). Through genome-wide bioinformatic identification, we systematically compared the abundance, density, structural categories, repeat motifs, chromosomal distribution, and pathway enrichment analysis of SSR-containing genes in CDS. Furthermore, we performed synteny analysis and evaluated population genetic diversity. A total of 2509 SSRs in CDS were identified in the FMD, with a relative density of 62.61 loci/Mb. Trinucleotide SSRs were overwhelmingly dominant (88.46%) in the FMD. Notably, the FMD exhibited the highest relative abundances of both tetranucleotide and pentanucleotide repeats among the six species (2.37 and 2.18 loci/Mb, respectively), with pentanucleotide abundance approximately 5.6- to 9.1-fold higher than that of the other species. Chromosomal mapping revealed the highest SSR density in CDS regions on chromosome 27, while SSR-containing genes exhibited a heterogeneous pattern characterized by localized clustering. Synteny analysis demonstrated relatively conserved syntenic relationships between the FMD and goat, sheep, and cattle, with moderate conservation also observed with red deer and white-tailed deer, suggesting that SSR-containing genes in ruminants may remain highly conserved during chromosomal rearrangements. GO and KEGG analyses indicated that SSR-containing genes across all species were predominantly enriched in transcriptional regulation, RNA processing, and signal transduction pathways. Specifically, the FMD showed enrichment patterns associated with hypoxia response, mRNA processing, and epigenetic regulation, which may reflect lineage-specific transcriptional patterns, though the functional involvement of these SSRs remains to be experimentally validated. In addition, the five primer pairs screened in this study exhibited high polymorphism, with a mean polymorphism information content (PIC) of 0.93. The observed heterozygosity (Ho) was significantly lower than the expected heterozygosity (He), and the mean inbreeding coefficient (FIS) was 0.57, indicating heterozygote deficiency and an elevated risk of inbreeding in this captive FMD population. Collectively, our findings provide preliminary insights into the conserved patterns of microsatellite evolution and lineage-specific divergence in ruminants, offering a reference framework for comparative genomics and adaptive evolution research, as well as practical molecular markers for genetic management of captive populations.

Veterinary Sciences doi: 10.3390/vetsci13080807

Authors: Vicky Fachinger Isabel Steiner Scott A. Callison Hubert Gantelet Kathrin Lillie-Jaschniski Thomas Lewiner Giovani Trevisan

The classification systems currently in use for Streptococcus suis are primarily based on capsular polysaccharide (CPS) serotyping or multi-locus sequence typing (MLST). A genetic typing system based on the conserved Mac-1 domain of the IdeSsuis gene, which encodes an IgM-cleaving protease, could complement these approaches. Specifically, in this study, a genotyping system based on Mac-1 sequence homology was established, qualitative SYBR Green real-time PCR (qPCR) assays for standardized genotype classification were developed, and the global and European distribution of genotypes were analyzed. The Mac-1 classification system identified “two genotypes”, one of which has “two subgenotypes”, based on 427 unique sequences obtained from 4161 global sequences. GT1.1 was the most prevalent, having an amino acid similarity of 97.1%, followed by GT1.2 98.1%, and GT2 91.4%. The qPCR approach enabled reproducible, cross-laboratory-accessible typing. Among 5479 Streptococcus suis European isolates, 64% were GT1.1, 4.5% were GT1.2, and 29.6% were GT2, with a trend toward increasing GT1.1 detection rates between 2018 and 2023. Genotyping classification could complement current Streptococcus suis classification approaches to better support vaccine design and disease control strategies.

Veterinary Sciences doi: 10.3390/vetsci13080805

Authors: Min Liu Zeming Song Shenghong Zhang Hehe Liu Fushi Li Renxia Yin Xiaoe Xiang Lian Li

This study evaluated the effects of dietary supplementation with a commercial lactoferrin–osteopontin complexes preparation (LF-OPN; LF:OPN mass ratio 8:1; 240 mg/kg body weight) on growth, immune indices, antioxidant status, intestinal barrier-related markers, fecal microbiota, short-chain fatty acids (SCFAs), peripheral lymphocyte subsets, and the whole-blood transcriptome of growing British Shorthair kittens. Sixteen kittens were randomly assigned to either a basal diet group (CON) or a group supplemented with LF-OPN complexes (LFT) for 28 days. LF-OPN supplementation did not significantly affect body weight or average daily gain. On D28, the LFT group had a higher serum immunoglobulin M concentration and a lower tumor necrosis factor-α concentration (p < 0.05). Serum superoxide dismutase and glutathione peroxidase activities and total antioxidant capacity were higher, whereas malondialdehyde was lower (p < 0.05) in the LFT group (p < 0.01). Fecal 16S rRNA gene sequencing revealed no significant difference in alpha diversity; howerer, LEfSe identified selective taxonomic difference, including higher relative abundances of Holdemanella and Erysipelotrichaceae and lower relative abundances of Enterobacteriaceae and Escherichia-Shigella in the LFT group. Fecal short-chain fatty acid concentrations were unchanged. Flow cytometry revealed a higher proportions of peripheral CD4+ lymphocytes and a higher CD4+/CD8+ ratio in the LFT group (p < 0.01), whereas the CD8+ proportion was unchanged. Whole-blood RNA sequencing identified 283 differentially expressed genes (199 upregulated and 84 downregulated), with enrichment of immune-related pathways including hematopoietic cell lineage, antigen processing and presentation, cytokine–cytokine receptor interaction, NF-κB signaling, and IL-17 signaling, but these findings were not independently validated. The tested LF-OPN preparation was well tolerated and was associated with elevated antioxidant status, altered immune indices, and selective modulation of the fecal microbiota in growing kittens. The study does not establish synergy or identify the individual contributions of either protein.

Veterinary Sciences doi: 10.3390/vetsci13080806

Authors: Mengting Zuo Pei Zhu Xi Yang Shuangshuang Zhai Peilin Yang Yi Zhou Lei Tan Jun Yao

Vector-borne pathogens, predominantly transmitted through arthropod vectors, present a major threat to both livestock production and public health. Melophagus ovinus is a significant disease vector widely distributed across various pastoral regions in China. However, there has been limited research on the prevalence of vector-borne pathogens within M. ovinus populations in Qinghai Province, China. In this study, a total of 433 M. ovinus specimens were collected from Haixi and Hainan prefectures in Qinghai Province between 2024 and 2025. After identifying the parasites via morphological and molecular approaches, the specimens were grouped into 84 pools and tested for the presence of three vector-borne pathogens using PCR techniques. The findings indicated that all collected specimens exhibited morphological features consistent with M. ovinus, and the 18S rRNA gene sequences of the selected parasites (n = 7) demonstrated sequence similarities ranging from 99.1% to 100% relative to reference M. ovinus strains. Among the 84 M. ovinus pools analyzed, 68 tested positive for Bartonella spp. DNA; subsequent genetic characterization identified these isolates (n = 15) as Bartonella melophagi. Additionally, Rickettsia spp. DNA was detected in 18 pools (21.43%), genetic characterization based on the OmpA gene (n = 6) identified all as Rickettsia conorii. No DNA from Babesia spp. or Theileria spp. was detected in any of the pool. Collectively, this investigation elucidates the epidemiological characteristics of three vector-borne pathogens among M. ovinus populations in Qinghai, China, and provides the first evidence of a high prevalence of both B. melophagi and R. conorii, emphasizing the necessity for targeted prevention and control measures.

Veterinary Sciences doi: 10.3390/vetsci13080804

Authors: Weldeab Solomon Ghebrezgabher Guodong Song Jiazhen Ge Yijian Liu Pengcheng Gao Renge Li Fuying Zheng Yuefeng Chu

Brucellosis is a zoonotic disease causing significant economic losses, and necessitating standardized diagnostic kits for effective surveillance. The objectives of this study are to investigate brucellosis serosurveillance in ruminants, and to evaluate three serological tests—Rose Bengal Plate Tests (RBPTs), Serum Tube Agglutination Tests (STATs), and Enzyme-Linked Immunosorbent Assays (ELISAs)—each from three manufacturers. A total of 5797 serum samples (5260 sheep, 83 bovines, and 454 spotted deer) were initially screened with the RBPT, and 54 RBPT-positive selected samples were subsequently analyzed by the STAT and ELISA. RBPT screening showed significant variability: kit A (0.86%) showed significantly lower positivity than kits B and C (1.45% and 1.43%, p < 0.05). Among 54 RBPT-positive samples, STAT positivity from manufacturers A, B, and C was 61.11%, 70.37%, and 66.67%, respectively, with no significant differences (p > 0.017 in pairwise comparisons). All ELISAs consistently outperformed STATs. Kit B (competitive ELISA, 90.74%) had significantly higher positivity than kits A (indirect ELISA, 77.78%) and C (competitive ELISA, 74.07%) (p < 0.017 in pairwise comparisons). Key implications: Results from different manufacturers are not directly comparable, and concurrent use of different techniques on the same sample can yield discrepant outcomes. This underscores the urgent need for diagnostic kit standardization in brucellosis prevalence studies.

Veterinary Sciences doi: 10.3390/vetsci13080803

Authors: Dan Anghelinici Mihai Musteata

Quantitative electroencephalography (qEEG) converts the raw EEG signal into reproducible numerical descriptors (spectral power, hemispheric symmetry, coherence and signal complexity) and has emerged as a candidate translational biomarker linking human and canine neurology. This integrative review examined the diagnostic, prognostic, pharmacological and translational value of qEEG, with emphasis on functional connectivity, and assessed the comparability of the dog as a natural model of human disease. Seventy-five studies were included, spanning epilepsy, acute brain injury, neurodegeneration, rehabilitation and paroxysmal disorders. In both species, epilepsy was consistently associated with altered spectral power and with reduced or reorganized coherence, and interictal abnormalities were demonstrable even in the absence of visible epileptiform discharges. Dogs reproduced the human patterns closely: phenobarbital induced the spectral redistribution predicted by human pharmaco-EEG data, canine cognitive dysfunction reproduced the slowing and the sleep-architecture changes described in Alzheimer’s disease, and a single machine-learning pipeline classified human and canine recordings with comparable accuracy. Conversely, acute brain injury remains virtually unexplored in the dog, and no canine normative database comparable to the human ones is yet available. The evidence was limited by heterogeneous acquisition protocols, small samples and scarce longitudinal veterinary data. qEEG, and coherence in particular, appears to be a promising cross-species biomarker of network dysfunction and supports the dog as a translational platform, although standardized validation remains necessary.

Veterinary Sciences doi: 10.3390/vetsci13080802

Authors: Baichuan Ma Jiaxue Wang Hui Tao Bing Han Zhenlong Wang Yongli Zhang Jinquan Wang Xiumin Wang

Wle vanillin is widely used as a flavoring agent in food products, its potential as a non-antibiotic feed additive for broilers remains unexplored. This study primarily investigated the effects of dietary supplementation of vanillin on antioxidant activity, anti-inflammatory response, immune function, and gut microbiota composition in broilers. One-day-old male broilers were randomly assigned to five groups: a basal diet group, a positive control group (basal diet + 100 mg/kg chlortetracycline (CTC)), and three vanillin-supplemented groups (0.1%, 0.2%, or 0.4%) for 42 days. Results showed that vanillin improved antioxidant capacity, with elevated serum superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity and reduced malondialdehyde (MDA) levels. Broilers receiving vanillin supplementation showed lower levels of pro-inflammatory cytokines (interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α)) and higher interleukin-10 (IL-10) expression compared with controls. Additionally, serum immunoglobulin A (IgA) and immunoglobulin M (IgM) levels were also greater in the vanillin group. It also enhanced intestinal health by increasing the duodenal villus height-to-crypt depth ratio, improving intestinal morphology, raising the Firmicutes-to-Bacteroidetes ratio, and promoting Lactobacillus proliferation compared with CTC. Notably, growth performance data were exploratory in nature; vanillin was associated with numerical improvements in average daily gain (ADG) and feed conversion (F/G) ratio. These preliminary findings suggest that vanillin requires further investigation as a potential feed additive.

Veterinary Sciences doi: 10.3390/vetsci13080801

Authors: Mengshi Chen Rongchang Liu Xin Yang Chuchu Duan Xiaozhen Yu Liyun Zhuang Tingting Dai Haiyu Chen Zehua Jin Zuchen Song Xintian Zheng

Gyrovirus homsa1 (GyH1) causes immunosuppression and multi-organ damage in young poultry. The VP1 capsid protein (VP1) is a key target for subunit vaccine development; however, recombinant VP1 alone elicits limited immunogenicity and requires an efficient delivery system. Here, we developed a liposomal formulation associated with the GyH1 VP1 protein (LNP-VP1) and evaluated its immunogenicity and preliminary safety in specific-pathogen-free chickens. The prepared LNP-VP1 had a mean particle size of 230.11 ± 5.95 nm, a polydispersity index of 0.204 ± 0.023, a zeta potential of −36.55 ± 0.99 mV, and an apparent encapsulation efficiency of 84.03% ± 2.10%. In addition, immunization of chicks with LNP-VP1 induced robust antigen-specific IgG responses and significantly enhanced the infiltration of CD4+ and CD8+ T cells in the spleen. Furthermore, cytokine analysis revealed upregulation of IFN-γ, IL-4, and IL-17 levels following vaccination. Importantly, no vaccine-associated histopathological changes were observed in major immune or metabolic organs after immunization. Overall, LNP-VP1 effectively elicited both humoral and cellular immune responses with a favorable safety profile, indicating its potential as a candidate vaccine against GyH1 and providing experimental evidence supporting lipid nanoparticles as a delivery platform for avian subunit vaccines.

Veterinary Sciences doi: 10.3390/vetsci13080800

Authors: Jennifer Hiekel Christoph Kühnle Klaudia Zofia Blaszczyk

A 13-year-old German Riding Pony mare was referred for evaluation of progressive respiratory noise and mixed dyspnoea, which rapidly deteriorated to respiratory distress. Clinical examination revealed marked rattling inspiratory and expiratory tracheal sounds and severe, rattling, vesicular lung sounds. Arterial blood gas analysis demonstrated moderate hypoxaemia (partial pressure of arterial oxygen 76 mmHg at rest). Upper airway endoscopy identified a pink, cauliflower-like pedunculated mass originating from the dorsal tracheal wall approximately 15 cm cranial to the tracheal bifurcation, causing an estimated 60% luminal obstruction. The mass was successfully removed via transendoscopic electrosurgical excision. However, complete tumour-free margins were not achieved. Histopathological examination revealed the diagnosis of leiomyosarcoma. Following surgery, the mare exhibited rapid clinical improvement. Peri-operative treatment included antimicrobial, anti-inflammatory, and bronchodilatory therapy. A follow-up endoscopy one month after surgery showed adequate healing of the surgical site. A small, vascularized tissue structure within the scar tissue, considered suspicious for residual tumour growth, was identified. However, no significant progression was observed during a follow-up examination seven months postoperatively. This case highlights tracheal leiomyosarcoma as a rare differential diagnosis in horses with respiratory distress and describes transendoscopic electrosurgical excision as a feasible therapeutic option.

Veterinary Sciences doi: 10.3390/vetsci13080799

Authors: Shuaishuai Wu Mohamed Tharwat Abd Ullah Nourhan Nassar Abdulrahman A. Alkheraif Muhammad Zahoor Khan

Dairy goats sustain milk production across marginal environments worldwide, yet genetic improvement of milk yield and composition remains slower than in cattle, constrained by smaller reference populations and limited functional validation. This review consolidates current evidence on the genes and polymorphisms governing milk production traits in dairy goats. Whole-genome resequencing and selection-signature scans repeatedly recover a lipogenic–somatotropic core (ACSS2, DGAT2, GHR, VPS13C, PRPF6), whereas genome-wide association studies (GWASs) confirm the casein cluster (CSN1S1, CSN1S2, CSN2, CSN3) as the most reproducible determinant of protein content and DGAT1 as the principal fat-content gene. Transcriptomic, single-cell, and metabolomic analyses, together with functional dissection in goat mammary epithelial cells, resolve the regulatory networks—SREBP1, PPARG, ELF5, and non-coding RNAs—that translate genotype into phenotype. Integrating multi-omic evidence with expanded, well-phenotyped populations is essential to convert this expanding catalog into precise marker-assisted and genomic selection for milk yield and quality.

Veterinary Sciences doi: 10.3390/vetsci13080798

Authors: Qiuyan Huang Qiqi Ma Shiyu Yang Yanling Wang Bin Zheng Weixia Ji Xingxing Song Xin Zhang Xun Li Xiaoye Wang Sutian Wang Chuanhuo Hu

Heat stress (HS) impairs boar reproductive capacity via damaging Sertoli cells (SCs) and disrupting the integrity of the blood–testis barrier (BTB). This present study investigated the protective effects and underlying molecular mechanisms of PTD-FNK against HS-induced injury in porcine SCs. A HS model (43 °C, 1 h) was established, and 0.1 nM was determined as the optimal working concentration of PTD-FNK. Results showed that PTD-FNK effectively reversed HS-induced BTB disruption by restoring the expression levels of tight junction proteins (Claudin-1, Occludin, ZO-1, and Cx43) to baseline levels (p < 0.05). Concurrently, PTD-FNK alleviated HS-induced oxidative stress by enhancing total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) activity, while reducing malondialdehyde (MDA) content (p < 0.05). It also suppressed HS-triggered apoptosis by downregulating the expression of pro-apoptotic proteins (Caspase-3/8/9 and Bax) and upregulating the anti-apoptotic protein Bcl-2 (p < 0.05). Furthermore, PTD-FNK maintained cellular homeostasis by regulating mitochondrial dynamics—restoring mitochondrial membrane potential (MMP) and balancing the expression of fusion-related protein Mfn1 and fission-related protein Drp1.Transcriptomic and LY294002 experiments confirmed PTD-FNK exerted effects via the PI3K/AKT pathway, modulating ID3, H2AX, DDIT3, CDKN1C, RAD51, and TP53. Thus, PTD-FNK protects SCs from HS through multi-target regulation dependent on PI3K/AKT, providing a novel strategy for boar reproductive health under HS.

Veterinary Sciences doi: 10.3390/vetsci13080797

Authors: Wantanee Tommeurd Vijittra Naithong Jullada Chootip Kingkarn Boonsuya Seeyo Porntippa Lekcharoensuk

Foot-and-mouth disease (FMD) is one of the most infectious viral diseases of cloven-hoofed animals. The crucial strategy for FMD control is the combination of animal quarantine and a rapid onsite diagnostic assay capable of differentiation of infected from vaccinated animals (DIVA). Herein, we have developed a lateral flow immunoassay to detect specific antibodies against FMDV non-structural protein (NSP), based on recombinant mu3ABC produced by E. coli as an antibody detector, the so-called mu3ABC strip test. The performance of this test was examined using 366 field serum samples from cattle, pigs, and goats and compared with the commercial ELISA kit, resulting in a diagnostic sensitivity (DSn) and specificity (DSp) of 98.05% and 93.16%, respectively. Additionally, efficacies of this test and commercial ELISA were determined using 200 reference bovine serum samples (100 positives, 100 negatives) classified by the Regional Reference Laboratory for Foot-and-Mouth Disease in South East Asia (RRL). The result reveals that the DSn and DSp of the test were 78% and 91%, respectively, while those for the commercial ELISA kit were 69% and 100%, respectively. The developed mu3ABC strip tests are effective and can be used to differentiate the infected animals in parallel with the sero-surveillance ELISA test.

Veterinary Sciences doi: 10.3390/vetsci13080796

Authors: Jianqing Zhao Wei Wang Yuzhu Guo Shuyu Shi Yuanpan Mu Jiayidaer Kamalibieke Chenbo Shi Lei Shi Jun Luo

Genetic background can shape metabolic phenotypes and may contribute to differences in health robustness among livestock breeds. This study investigated Awassi-associated serum metabolic characteristics in dairy sheep by integrating untargeted metabolomic profiling with farm-level morbidity information. Serum samples were collected from 36 adult sheep representing six genetic groups raised under common feeding and management conditions, Awassi, East Friesian, Hu, and three Awassi–East Friesian–Hu crossbred groups, with six animals per group and an equal number of males and females. Following data preprocessing and normalization, 2284 metabolic features were retained for comparative analysis. Principal component analysis revealed marked differences in serum metabolic profiles among the genetic groups, with Awassi sheep displaying a distinct metabolic pattern relative to the purebred and crossbred populations. Across five comparisons between Awassi and the other genetic groups, 17 metabolic features were consistently more abundant in Awassi sheep. These features were mainly associated with lipid mediator metabolism, glycerophospholipid metabolism, aromatic and indole-related metabolism, and host–microbial co-metabolism. A metabolic signature derived from the Awassi–East Friesian comparison differed significantly among the six groups and was positively associated with Awassi ancestry at the individual level. However, the two crossbred groups with 50% Awassi ancestry did not show a simple intermediate metabolic pattern, suggesting that crossbred metabolic phenotypes may involve non-additive genetic effects. Farm records showed lower total morbidity in Awassi and the three crossbred populations than in the East Friesian and Hu populations. Because the morbidity data were summarized at the population level and were not individually matched to the metabolomic samples, they were interpreted as supporting production context rather than direct metabolite–disease associations. Collectively, these results reveal a distinctive Awassi-associated serum metabolic signature and provide new insights into breed- and ancestry-related metabolic variation in dairy sheep.

Veterinary Sciences doi: 10.3390/vetsci13080795

Authors: Alejandro Fenollar María A. Ferrús Pablo Catalá-Gregori Vicente Tallá-Ferrer Miguel García-Ferrús Ana Isabel Jiménez-Belenguer

Enterococcus faecalis and Enterococcus faecium are commensal bacteria of poultry but also important opportunistic pathogens and key indicators for antimicrobial resistance (AMR) surveillance within a One Health framework. However, data on the occurrence and antimicrobial resistance of enterococci in breeding hens remain limited despite their critical role in the poultry production pyramid. This study evaluated the prevalence, antimicrobial resistance profiles, and resistance genes of Enterococcus faecalis and Enterococcus faecium isolated from two commercial breeding hen farms in eastern Spain. A total of 330 isolates were obtained from different production stages and sample types. Species identification was performed by multiplex PCR, antimicrobial susceptibility was assessed using disk diffusion according to Clinical and Laboratory Standards Institute (CLSI) guidelines, and resistance genes (erm and tet) were detected by PCR. A significant age-related shift from E. faecalis in early production stages to E. faecium in adult hens was observed. High resistance rates were detected for tetracycline (up to 95.2%) and erythromycin, whereas resistance to critically important antimicrobials such as vancomycin, aminoglycosides, and ampicillin was rare or absent. Significant differences between farms were identified in both antimicrobial resistance levels and the prevalence of multidrug resistance. The most prevalent resistance genes were ermB, tetL, and tetM, frequently co-occurring. This study shows that breeding hens constitute a relevant reservoir of AMR within poultry production systems. Species dynamics and resistance patterns appear to be influenced by age and farm-related factors. These findings underscore the need for continuous AMR surveillance integrating phenotypic and genotypic approaches from a One Health perspective.

Veterinary Sciences doi: 10.3390/vetsci13080794

Authors: Tayyaba Ashiq Kexin Yu Xiaoxiang Pan Babar Maqbool Shuying Zhong Zhaofeng Hou Yanhong Wang Penggang Liu

Toxoplasmosis is a zoonotic disease caused by the intracellular protozoan parasite Toxoplasma gondii, with marsupials recognized as particularly vulnerable hosts. This is a case report of the integrated investigation of fatal illness in a captive red kangaroo (Osphranter rufus). Multisystemic lesions with hemorrhages and congestion of the lungs, liver, spleen, kidneys, heart, and intestines were detected during gross necropsy. Hematological analysis demonstrated leukopenia (2.90 × 109/L), neutropenia (1.19 × 109/L), lymphopenia (1.53 × 109/L), and increased hematocrit (61.2%). The serum alanine aminotransferase level was very high (216 U/L), and urea nitrogen and creatinine were elevated, indicating involvement of the liver and kidneys. A bacteriological investigation based on the sequences of 16S rDNA revealed the presence of Neisseria weaveri from the heart and Staphylococcus arlettae from the kidney; no Salmonella spp. or Clostridium perfringens were isolated. The DNA of T. gondii was detected by PCR in various tissues, including spleen (low-level) and lung (high-level). Pathohistological examination of the spleen demonstrated lymphoid depletion and inflammatory infiltration without tissue cysts, tachyzoites, granulomatous inflammation, or multifocal necrosis. Characteristic lesions are absent, and there is no way to definitively attribute mortality to fatal toxoplasmosis, as the lung had the strongest PCR signal. Collectively, these results support PCR-based molecular detection of disseminated T. gondii DNA and emphasize the necessity of comprehensive histopathological correlation with all PCR-positive organs in future investigations.

Veterinary Sciences doi: 10.3390/vetsci13080793

Authors: Gan Li Qipeng Zhang Xiaojing Chen Huawei Li Ruining Wang Keshan Zhang Yaqiong Ye Mengmeng Zhao

As a highly contagious infectious disease in the swine industry, porcine reproductive and respiratory syndrome (PRRS) can cause reproductive disorders in sows and respiratory diseases in piglets, resulting in sustained economic losses. Nonstructural protein 10 (NSP 10), as one of the core components of the replication and transcription complex of PRRS virus (PRRSV), has both helicase and adenosine triphosphatase activities, and has the potential as a diagnostic marker. In this study, NSP10 recombinant protein was used as coating antigen, and an indirect ELISA method for PRRSV NSP10 detection was established by square matrix titration. After optimizing the assay parameters, high specificity (no reactions with other pig viruses), high sensitivity (able to detect antibodies even when the positive blood sample was diluted 1280 times), and excellent reproducibility (less than 10% variation) were shown. Therefore, the NSP10 indirect enzyme-linked immunosorbent assay (ELISA) developed in this study provides an effective technical means for enhancing PRRSV surveillance in swine herds, thereby establishing a solid foundation for the clinical diagnosis and comprehensive prevention and control of PRRSV.

Veterinary Sciences doi: 10.3390/vetsci13080791

Authors: Luciano Bianco do Amaral Eder Batalha Araujo Pedro Nacib Jorge-Neto Dominike Prediger Delazeri Janine de Camargo Mariana Groke Marques Ricardo Zanella

Semen quality is a critical factor for the success of artificial insemination programs in swine production, and is influenced by collection method, genetic lineage, collection interval and pharmacological interventions. This study evaluated the effects of Dinoprost-tromethamine (DP; Lutalyse®, Zoetis, Parsippany, NJ, USA), a prostaglandin F2α analog, on boar semen quality in a commercial stud, considering manual versus semi-automatic collection systems, four genetic lines and resting time between collections. A total of 653 boars (146 Large White, 210 Landrace, 282 Duroc and 15 Hybrid [Duroc × Pietrain]) were monitored over 12 months, resulting in 22,977 semen collections. DP was administered intramuscularly as a single dose of 1 mL per boar (6.71 mg of active compound; approximately 0.028 mg/kg body weight), 20 min before semen collection, exclusively to animals classified with low libido (scores 1–3), whereas boars with normal libido (scores 4–6) remained untreated. Semen parameters (ejaculate volume, sperm concentration, total motility and kinematic traits assessed by CASA) were analyzed using mixed-effects models including collection method, DP treatment, genetic line and their interactions. Semi-automatic collection increased ejaculate volume and total motility but resulted in lower sperm concentration compared with manual collection. DP administration increased sperm motility and sperm concentration while reducing ejaculate volume, with the greatest improvement in motility observed in Hybrid boars and more modest or nonsignificant responses in the other genetic lines. Longer collection intervals were associated with higher sperm concentration, whereas ejaculate volume and motility were not affected by resting time. These findings indicate that the impact of DP, collection method and collection interval on semen quality is breed-dependent and highlight the need to tailor reproductive management protocols according to boar genetics, libido and collection technology.

Veterinary Sciences doi: 10.3390/vetsci13080792

Authors: Maged A. Al-Garadi Rashed A. Alhotan Elsayed O. Hussein Mohammed M. Qaid Gamaleldin M. Suliman Esam H. Fazea Mohammed A. Al-Badwi Isiaka O. Olarinre

This study evaluated the effects of Fibrafid, a plant-based functional feed additive, on growth performance, carcass traits, meat quality, and intestinal histomorphometry of broilers fed standard or moderately nutrient-reduced diets. A total of 432 one-day-old Ross 308 chicks (initial body weight, 44.9 ± 0.30 g) were randomly allocated to a 2 × 3 factorial arrangement comprising two dietary nutrient densities (standard or diets with 1.5% lower metabolizable energy and 5% lower digestible amino acid density) and three Fibrafid inclusion levels (0, 1.5, or 2.5 kg/ton). Each treatment comprised six replicates of 12 birds (six males and six females) and was evaluated from day 1 to 24. Dietary nutrient reduction affected growth performance and breast meat quality pH and ultimate lightness values, whereas Fibrafid supplementation, particularly at 2.5 kg/ton, improved overall gain by 5.3%, grower-phase feed conversion ratio by 4.0%, EPEF by 8.08%, relative thymus weight by 64.8%, while reducing abdominal fat deposition by 27.1% relative to the unsupplemented control (p < 0.05). Several breast meat quality traits were also improved, including greater water-holding capacity and myofibril fragmentation index and lower drip and cooking losses (p < 0.05). Jejunal morphology was enhanced, as indicated by increased villus dimensions and goblet cell density (p < 0.05). Significant Diet × Fibrafid interactions were observed for grower performance, EPEF, overall feed intake, and several intestinal morphological variables (p < 0.05). In conclusion, Fibrafid supplementation, particularly at 2.5 kg/ton, was associated with improved growth performance, EPEF, selected carcass and breast meat quality traits, and jejunal morphology during the first 24 days of age. These findings suggest that Fibrafid may be a promising natural feed additive for broiler diets formulated at standard or moderately reduced nutrient densities. However, further studies extending to market age and evaluating nutrient digestibility, intestinal microbiota, blood biochemical indices, and economic returns are warranted to clarify its mechanisms and practical value under commercial production conditions.

Veterinary Sciences doi: 10.3390/vetsci13080790

Authors: Lucía Vázquez-Salgado Carmen López-Vázquez Isabel Bandín Sandra Souto

Quantifying the expression of immune-related genes is essential for evaluating vaccine efficacy. However, obtaining reliable RT-qPCR results requires the use of stable reference genes. In this study, the expression stability of eight candidate reference genes (actb1, actb2, eef1a, gapdh2, hprt1, ubq, 18S, and rps4) was evaluated in brain, kidney, and gut tissues from naïve and virus-infected juvenile sole. Gene stability was assessed through two statistical algorithms (geNorm and NormFinder) and the integrative tool RefFinder, and the results were integrated through RankAggreg for a robust consensus ranking. Among the tested genes, rps4 and ubq showed the highest stability across tissues and experimental conditions and were sufficient for accurate normalisation. The suitability of these genes was validated in vaccinated sole for normalisation of the expression of proinflammatory cytokines (il1β, il6, tnfα), supporting their recommendation as optimal reference genes. Conversely, actb1 and hprt1 were the least stable genes and compromised the reliable quantification of proinflammatory cytokines in vaccinated fish.

Veterinary Sciences doi: 10.3390/vetsci13080789

Authors: Dexin Zhao Haoliang Chai Linfeng Zhou Ruicheng Han Weiqi Peng Hongzhi Wu

Poultry nutrition simultaneously affects animal health, productive efficiency, product quality and the environmental burdens associated with feed production and manure management. Nutritional interventions may generate synergies among these outcomes, but they can also shift costs or environmental impacts across biological and supply-chain boundaries. This review critically evaluates low-protein diets, functional additives, feed enzymes, alternative protein sources, precision-feeding technologies and nutritional approaches to manure-emission mitigation. The evidence indicates that improved intestinal function and nutrient retention can reduce feed requirements and nutrient excretion, whereas excessive protein restriction, unstable additive responses, limited alternative-ingredient supply and poorly validated digital systems may compromise performance, product quality or economic feasibility. Environmental benefits are also sensitive to ingredient origin, processing, transport, crystalline amino acid and additive manufacture, coproduct allocation and life cycle assessment assumptions. To integrate these considerations, we propose a four-level conceptual decision framework comprising physiological adequacy, precision nutrient supply, constraint-specific functional intervention and whole-chain verification. The framework treats health, productive performance, product quality, economic feasibility and environmental impact as simultaneous decision criteria, while genotype, production stage, disease pressure, climate, ingredient availability and market objectives define the conditions of application. Standardized life cycle assessment and commercial validation are required to distinguish reproducible net benefits from improvements confined to individual indicators. This approach provides a structured basis for selecting poultry nutritional strategies that remain biologically effective, economically accessible and environmentally beneficial under practical production conditions.

Veterinary Sciences doi: 10.3390/vetsci13080788

Authors: Li Wang Yaqian Jin Guoli Zhang Xue Chen Hongrui Ren Lanjie Li Jingjing Tian Bayier Mengke Peiming Li Yongjia Ma Caolemeng Qiqige Wa Gao Guiqin Liu Ruiwen Fan

(1) Background: Anti-Müllerian hormone (AMH) is a key indicator for predicting ovarian reserve and fertility. Currently, B-mode ultrasound examination is the primary method for evaluating follicular development in Bactrian camels, although it has several limitations. This study aims to develop a colloidal gold strip based on AMH nanobody preparation, establishing an efficient, simple, and rapid detection method suitable for evaluating follicular development. (2) Methods: Two nanobodies specifically binding to AMH were screened and identified from an alpaca naive nanobody library using phage display technology. The nanobodies were expressed in the yeast expression vector (pMCO-AOXα) and purified by affinity chromatography on a nickel column. Based on the purified nanobodies, a colloidal gold immunochromatographic strip was constructed for detecting serum AMH levels. (3) Results: In this study, anti-AMH nanobodies (AMH-VHH-A1 and AMH-VHH-A10) were expressed and purified; these were successfully utilized to prepare colloidal gold immunochromatographic strips for detecting AMH in serum. The T-line intensity of the colloidal gold detection was positively correlated with the number of growing follicles obtained by B-mode ultrasound imaging. (4) Conclusions: The colloidal gold strips based on nanobodies against AMH established in this research provided an effective, simple, rapid, and on-site method for evaluating follicular development, thus supplying a reference for reproductive management in Bactrian camels.

Veterinary Sciences doi: 10.3390/vetsci13080787

Authors: Victoria Luño Clara Zabalo-Palomo Maite Olaciregui Beatriz de la Barreda Lydia Gil

This study was conducted to study the quality and DNA integrity of freeze-dried stallion sperm, focusing on the influence of sperm origin (ejaculate vs. epididymis) and seasonality (breeding vs. non-breeding season). Ejaculates were collected from six healthy stallions and epididymal sperm were obtained from 12 stallions via cauda epididymis perfusion during the reproductive season and non-reproductive season in both cases. Sperm samples were diluted in an extender composed of 0.1 M trehalose and BSA 12.5% in PBS, freeze-dried, and stored at 4 °C for at least 3 months. After rehydration, sperm plasma membrane integrity and DNA integrity were analysed using eosin–nigrosin staining and a chromatin dispersion test (SCDt Halomax®), respectively. Differences in DNA damage were found to be a function of sperm origin. The percentage of DNA fragmentation in ejaculated sperm was significantly higher than in epididymal sperm. However, when evaluating the effect of the season (breeding vs. non-breeding time), DNA integrity did not differ between seasonal groups. In relation to sperm plasma membrane integrity, no differences were found between the experimental groups. Changes in sperm DNA integrity were observed, independently of season, between epididymal and ejaculated stallion sperm after freeze-drying. This study identifies different factors that lead to improved sperm DNA quality after lyophilisation.

Veterinary Sciences doi: 10.3390/vetsci13080786

Authors: Antonella Di Paolo Rosario Liberti Lucia Anzalone Claudia Colabella Andrea Felici Anita Emilia Colombo Patrizia Bonfanti Pamela Floris Martina Pellegrini Giulia Vita Monica Cagiola

Background: Veterinary autogenous vaccines (AVs) are emergency tools produced in a short timeframe from bacterial/viral/fungal isolates when commercial vaccines are not available and undergo strict quality assurance and safety tests before release. In accordance with the European Directive 2010/63/EU and the 3R principles, we evaluated for the first time the possible substitution of the final toxicity in vivo test (abnormal toxicity test) with an alternative cell-based and embryo-based assay. Methods: The five most requested different autogenous matrices (adopted to produce a customizable number of lots requested from customers) were tested in vitro in parallel for residual toxicity by in vitro fish embryo acute toxicity (FET) test and cytotoxicity assay (MTS) using serial dilutions, together with the authorized in vivo test. Results: Statistical analysis revealed different outcomes between the two in vitro methods, highlighting the greater discriminatory power of the MTS assay compared to the other, with results more like the observed in vivo ATT outcome (pass/negative). Conclusions: In this preliminary first dataset, FET appeared poorly compatible with several complex autogenous vaccine matrices, while the L929 MTS assay yielded interpretable viability thresholds for all tested matrices. These findings support further qualification of MTS as part of an integrated non-animal batch characterization strategy, but they do not yet establish replacement of the in vivo test for veterinary autogenous vaccine quality control.

Veterinary Sciences doi: 10.3390/vetsci13080785

Authors: Ahmad A. Aboragah Abdulrahman S. Alharthi Hani H. Al-Baadani

(1) Background: The global transition away from antibiotic growth promoters (AGPs) requires effective, sustainable alternatives to maintain broiler performance and intestinal homeostasis. This study evaluated the effects of dietary wild thyme powder, administered alone or combined with a prebiotic or probiotic, as potential AGP alternatives on growth performance, serum oxidative status, immune response, cecal microbiota composition, short-chain fatty acid (SCFA) production, ileal histomorphometry, and intestinal barrier integrity in broilers. (2) Methods: Five hundred forty-six male Ross 308 chicks were assigned to seven treatments (n = 13 replicates/group) for 28 d: T1 (basal diet); T2 (0.5 g/kg antibiotic Neoxyval); T3 (1.0 g/kg thyme); T4 (0.75 g/kg prebiotic); T5 (0.50 g/kg probiotic); T6 (prebiotic + thyme); and T7 (probiotic + thyme). Growth performance was measured, and one bird from each replicate was randomly sampled at 28 d for multi-endpoint biological analyses. (3) Results: Birds that received T3 to T7 had increased final body weight and weight gain. Cumulative feed conversion ratio was improved in the T3, T5, and T7 groups, with T7 displaying numerical superiority over individual additive groups without reaching statistical synergy. All-natural supplements (T3, T5, T6, T7) lowered serum malondialdehyde and enhanced interferon-γ. Birds receiving T6 showed increased tumor necrosis factor-α, whereas birds receiving T3 and T4 showed reduced levels. Probiotic-inclusive diets (T5 and T7) significantly increased cecal propionic acid, total SCFAs, and ileal villus surface area. Natural additive treatments broadly promoted cecal eubiosis by increasing Lactobacillus spp. while decreasing Clostridium spp. Crucially, combining thyme with prebiotics (T6) or probiotics (T7) synergistically upregulated ileal tissue occludin concentrations. (4) In conclusion, under the conditions of this trial, dietary supplementation with wild thyme powder, a MOS/β-glucan prebiotic, and a Bacillus licheniformis probiotic, particularly the thyme + probiotic combination, was an effective regimen for supporting growth performance, cecal eubiosis, and ileal barrier integrity in healthy broilers. However, these findings are limited to a single dose level (1.0 g/kg thyme), a single plant source, and non-challenged conditions; further research incorporating dose–response evaluations, disease-challenge models, and diverse phytogenic origins is necessary to confirm broader field applicability.

Veterinary Sciences doi: 10.3390/vetsci13080784

Authors: Annamária Kiss Gábor Lorászkó Gabriella Korsós Dorottya Bozsó Zsombor Wagenhoffer Kinga Fodor

Animal abuse and substandard animal care may be difficult to report when members of the public are uncertain about the seriousness of their observations, the competent authority, or the information needed for further assessment. This study aimed to develop and preliminarily evaluate a structured online reporting interface for suspected animal abuse and animal welfare concerns in Hungary. Two public online questionnaires assessed reporting-related knowledge, experiences, and preferences, and a separate professional questionnaire examined the experiences of actors involved in animal protection procedures. Based on these findings, the Hungarian legal context, and international reporting models, a browser-based prototype was developed and professionally tested by staff members of the National Food Chain Safety Office. Public respondents significantly differentiated between suspected criminal animal cruelty and substandard animal keeping: the police were selected more often for animal cruelty cases, whereas the notary was selected more often for substandard keeping cases (p < 0.001). Substandard keeping conditions were observed significantly more frequently than direct animal abuse (p < 0.001). Professional testing indicated that the prototype was understandable, transparent, and potentially useful, although further refinement, institutional integration, and field validation are required.

Veterinary Sciences doi: 10.3390/vetsci13080783

Authors: Jingyi Gao Yuhan Ding Aoxiang He Wanyan Zhang Huaqun Chen

Bovine mastitis, caused by both contagious and environmental pathogens, represents a major infectious disease burden in the global dairy industry. Antibiotics remain the primary treatment option, but their effectiveness is limited by the blood–milk barrier, drug residues, and the growing threat of multidrug-resistant bacteria. Among the various alternatives, phage therapy has drawn particular attention due to its specificity, ability to disrupt biofilms, low impact on commensal flora, and self-replication at infection sites. In this review, we summarize recent advances in the isolation of lytic phages targeting major mastitis-causing pathogens, their bactericidal mechanisms, and their performance in vitro and in vivo. We also discuss key obstacles to clinical translation including formulation stability, narrow host range, and safety concerns. Moreover, the potential strategies to overcome these issues are explored. This review provides a useful reference for research on phage therapy against bovine mastitis.

Veterinary Sciences doi: 10.3390/vetsci13080782

Authors: Ahmad Ali Mohammad Adil Bilal Ahmad Muhammad Ilyas Rakhshanda Rani Uzair Alam He Hongsu Zhang Hui Sun Zhihua

Antimicrobial resistance (AMR) is a major One Health challenge driven by antimicrobial misuse in human medicine, veterinary practice, animal production, and polluted environments. Migratory birds move among wetlands, farms, wastewater-affected habitats, landfills, and coastal ecosystems and may acquire and redistribute antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) across ecological and political boundaries. This perspective synthesizes evidence on exposure sources, bacterial reservoirs, resistance determinants, cross-species interfaces, and surveillance priorities while explicitly distinguishing four claims: detection or carriage, persistence in individual birds, redistribution along migratory routes, and onward transmission to recipient hosts or environments. Published studies report multidrug-resistant Escherichia coli, Klebsiella pneumoniae, Salmonella spp., Enterococcus spp., and Campylobacter spp., with determinants including blaCTX-M, blaTEM, blaNDM, mcr, tet, sul, and qnr genes. The eight evidence groups summarized here constitute an illustrative, non-comprehensive selection; they are predominantly observational surveys or screenings, and none reconstructs a complete source–bird–destination–recipient transmission chain. Taxon-specific ecology modifies exposure: gulls and storks frequently exploit refuse and wastewater, waterfowl and shorebirds connect aquatic habitats, whereas passerines often reflect more local point-source contamination. Current evidence therefore supports migratory birds primarily as mobile sentinels and opportunistic carriers of anthropogenic AMR, while acknowledging possible natural or ancestral resistance in avian-associated microbiota. Future surveillance should combine longitudinal sampling, baseline cohorts such as pre-migratory nestlings, paired bird–water–soil–sediment sampling, whole-genome sequencing, plasmid profiling, telemetry, environmental DNA, wastewater-based epidemiology, and interoperable veterinary reporting. Practical mitigation requires antimicrobial stewardship, wastewater and landfill control, farm biosecurity, and coordinated veterinary, environmental, and public-health action.

Veterinary Sciences doi: 10.3390/vetsci13080781

Authors: Akihiro Ohnishi Mio Takeuchi Ryo Sakaguchi Runa Iwata Taketoshi Asanuma Yoshiki Itoh

Noncontact camera-based detection of respiratory motion may support low-burden surveillance of hospitalized dogs, particularly at night. This pilot study evaluated the feasibility of the InsectEye image-analysis application for detecting visually identifiable respiratory motion in selected infrared nighttime video segments. An initial parameter optimization was performed using 13 one-minute video segments from nine dogs. The selected analysis conditions were subsequently evaluated in Experiment 1 using five 5 min video recordings from four dogs. Experiment 2 further assessed respiratory rate estimation using the same validation dataset. InsectEye-detected events were compared with visually confirmed respiratory motion using time-synchronized review. Two criteria were assessed: Both-only, requiring detection of both inspiratory- and expiratory-related motion, and Both+Either, accepting detection of either or both phases. Detection performance improved when the threshold was reduced from 10 to 8, particularly with the 256 × 120 mesh. The 256-120_15_8 condition yielded the highest mean detection rate (87.93 ± 1.87%), fewer false-negative events, and shorter continuous non-detection duration, although false-positive detections increased. The Both+Either criterion performed better than the Both-only criterion. These findings support the feasibility of camera-based respiratory motion detection under selected favorable conditions, but larger studies using physiological respiratory reference methods are required before clinical monitoring or alerting applications can be established.

Veterinary Sciences doi: 10.3390/vetsci13080780

Authors: Haifa Ali Alqhtani Tahani M. I. Al-Hazani Ahmed El Sayed Ahmed Ateya Ahmed H. Ghonaim Rowa K. Zarah Fatmah A. Safhi Adel Almubarak Hussein Babiker Rasha yassin Elkhidr Wael M. El-Deeb Ahmed Magzoub Khalid Mayyadah Abdullah Alkuwayti Mohamed Marzok

Identifying physiological changes during the transition period is essential for improving the health and productivity of dairy goats. This study evaluated hematological, biochemical, hormonal, oxidative stress, and molecular alterations in Shami goats during the pre-pregnancy, late pregnancy, and early lactation periods. Eighty clinically healthy goats were examined, and blood samples were analyzed for hematological indices, metabolic and hormonal profiles, oxidative stress biomarkers, and relative expression of genes associated with energy metabolism, antioxidant defense, inflammation, and autophagy. Late pregnancy was characterized by significant (p < 0.05) increases in red blood cell count (RBCs), hemoglobin concentration (Hb), neutrophils, albumin, globulin, urea, insulin-like growth factor-1 (IGF-I), and malondialdehyde (MDA), accompanied by decreased glucose, cholesterol, total protein (TP), antioxidant markers, total leukocyte count, packed cell volume, and monocytes. Early lactation was associated with higher non-esterified fatty acid, triiodothyronine (T3), and thyroxine (T4) levels. Genes involved in lipid mobilization and oxidation, ketogenesis, inflammation, cellular stress, and autophagy; sirtuin 1 (SIRT1), peroxisome proliferator-activated receptor alpha (PPARA), carnitine palmitoyltransferase 1a (CPT1A), 3-hydroxy-3-methylglutaryl-coenzyme a synthase 2 (HMGCS2), cluster of differentiation 36 (CD36), lipase E (LIPE), protein kinase amp-activated catalytic subunit alpha 1 (PRKAA1), solute carrier family 2 member 1 (SLC2A1), haptoglobin (HP), interleukin 6 (IL6), heat shock protein 70 (HSP70), heme oxygenase 1 (HMOX1), beclin 1 (BECN1), and autophagy-related protein 5 (ATG5) were significantly upregulated, whereas antioxidant- and glucose transport-related genes nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione peroxidase 1 (GPX1), catalase (CAT), thioredoxin (TXN), and solute carrier family 2 member 4 (SLC2A4) were downregulated during the transition period. The results indicate well-orchestrated metabolic and molecular adaptations that can be employed as biological indicators to track the physiological status of Shami goats. These findings fulfilled the study objective and identified potential biomarkers of the transition period in Shami goats.

Veterinary Sciences doi: 10.3390/vetsci13080779

Authors: Ayesha Sarwar Bilal Aslam Sulaiman F. Aljasir

Shiga toxin-producing Escherichia coli (STEC) contamination and proliferation in aquaculture and aquatic systems is worrisome for global food safety, as well as veterinary and public health. Traditionally linked with terrestrial ruminant reservoirs, aquaculture matrices, including farmed finfish, shellfish, culture water, and benthic organisms, are increasingly acknowledged as potential conduits for the dissemination of STEC. Herein, the review documented data concerning the prevalence, genomic composition, and ecological dynamics of STEC across various aquaculture environments across different regions of the globe. As a result, the persistence of virulence-associated genes (VAGs) and antibiotic-resistant genes (ARGs) in STEC within the aquaculture supply chain presents a considerable risk to global food security and public health, highlighting the urgent necessity for comprehensive “One Health” surveillance frameworks aimed at alleviating aquatic biosecurity challenges.

Veterinary Sciences doi: 10.3390/vetsci13080778

Authors: Ying Fu Zhiling Guo Huiyan Wang

Monoclonal antibodies are emerging as advanced therapies in companion-animal medicine, with approved or clinically evaluated applications in canine and feline osteoarthritis pain, canine atopic dermatitis, and selected infectious-disease and oncological settings. This critical narrative review integrates peer-reviewed studies, regulatory records, product information, and publicly available therapeutic-antibody sequence resources to examine target biology, species adaptation, Fc function, developability, and clinical implementation. Anti-NGF and anti-IL-31 products demonstrate the clinical value of long dosing intervals, whereas oncology and infectious-disease programmes remain subject to indication-specific efficacy and safety requirements. Canine and feline IgG subclasses, Fc-receptor and FcRn biology, incomplete feline framework resources, manufacturing economics, and owner-funded care all influence candidate design and deployment. Emerging bispecific antibodies, antibody–drug conjugates, VHHs, mRNA-based approaches, and AI-assisted strategies are most relevant when they address a defined veterinary use case. We propose a translational framework linking target validation, species-adapted engineering, species-matched assays, manufacturability, regulatory evidence, and real-world deployment.

Veterinary Sciences doi: 10.3390/vetsci13080777

Authors: Xianglong Zhang Xitong Guan Jiahui Cao Yuxuan Yan Yueyang Zhao Hongxiang Mao Lizhou Ma Lingling Huang Xiangfang Tang Shunjin Jiang Yang Li

This study evaluated the impact of fermented palm kernel meal (FPKM) on the lactation performance, blood biochemical indices, rumen microbiota, and metabolic functions of Holstein dairy cows, aiming to enhance the nutritional value of palm kernel meal (PKM) through solid-state fermentation. A 3 × 3 Latin square design was used, involving 12 multiparous Holstein cows (parity = 3; body weight = 625 ± 25.8 kg; days in milk = 103 ± 19.6 day(s); milk yield = 32.6 ± 1.58 kg/d) over three 28-day periods. Cows were randomly assigned to three isocaloric and isonitrogenous diets: a basal diet with wheat bran (WB group), a diet with wheat bran replaced by PKM (PKM group), and a diet with wheat bran replaced by FPKM (FPKM group). Solid-state fermentation improved PKM’s nutritional profile by reducing fiber and β-mannan content while increasing protein availability and ruminal degradability. Compared to the WB group, the PKM group showed lower dry matter intake, milk yield, and nutrient digestibility. In contrast, the FPKM group had higher DMI and milk yield than the PKM group, improved nutrient digestibility, and the highest energy-corrected milk yield due to increased milk protein and lactose production. The FPKM group also had higher concentrations of total volatile fatty acids, propionate, acetate, and microbial protein synthesis than the PKM group. Pro-inflammatory cytokines (tumor necrosis factor-α, interleukin-8) were elevated in the PKM group but were reduced to levels similar to the WB group in the FPKM group. Plasma immunoglobulin G levels were higher in both the FPKM and WB groups compared to the PKM group. The FPKM group also showed increased relative abundances of Prevotella, Fibrobacterota, and Verrucomicrobiota, while Bacillota and Ruminococcus were reduced compared to the WB group. Metabolomic profiling revealed that FPKM upregulated energy metabolism and inflammation-related pathways, increasing metabolites such as riboflavin and adenine and decreasing succinic acid and guanine compared to the PKM group. In conclusion, FPKM improved the feeding value of PKM-based material and showed more favorable responses than PKM, with generally comparable responses to WB, supporting its potential as an alternative feed ingredient for lactating dairy cows.

Veterinary Sciences doi: 10.3390/vetsci13080776

Authors: Yu He Fengyi Dong Long Jin Jiayi Ning Wuyang Liu Chengyue Feng Zhikai Zhu Han Sun Xiaoran Zhang Changyi Chen Luyao Bie Boxing Sun Hao Sun Chunyan Bai

The Jishen Black pig is a synthetic breed incorporating the genetic backgrounds of Chinese indigenous pigs and Western lean-type pigs. The objective of this study was to characterize phenotypic variation, estimate genetic parameters, and identify genomic loci associated with rib number (NR) and the total number of thoracolumbar vertebrae (NTLV) in Jishen Black pigs. NR, NTLV, and lumbar vertebra number (NLV) were measured in 389 pigs, and genotyping was performed using a 70K SNP chip. Genetic parameters were estimated using HIBLUP, and genome-wide association studies (GWAS) were conducted using the MLM and BLINK models in GAPIT v3.0. The mean values of NR, NTLV, and NLV were 14.88, 20.94, and 6.05, respectively, and their heritability estimates were 0.568, 0.493, and 0.140, respectively. GWAS identified a shared major association peak for NR and NTLV on SSC7 at 91.19–98.15 Mb, centered on VRTN and the adjacent ABCD4-LTBP2-AREL1-PGF linkage region, among which the loci at 96.12 Mb, 96.56 Mb and 97.79 Mb are NR-specific loci. Additional signals included MIB1 on SSC6 and MMRN2 on SSC14 for NTLV, together with NR-specific loci on SSC7 at 111.59 Mb and on SSC8. Database annotation indicated that several significant SNPs overlapped with records for rib number, carcass length, loin muscle area, backfat thickness, or teat number. This study identifies major genomic regions associated with vertebral-number traits in Jishen Black pigs and expands current knowledge of their genetic basis.

Veterinary Sciences doi: 10.3390/vetsci13080775

Authors: Hanan S. Al-Khalaifah Asmaa T. Y. Kishawy Rania M. S. El-Malt Wessam Youssef Walaa A. Habib Dalia W. A. H. Elged Wafaa M. Gad Eman A. Elalfy Mohammed E. E. Sayed Ahmed Hebatullah M. Abouelfadl Nanies S. E. Salim Marwa M. Fathi Doaa Ibrahim

Multidrug-resistant (MDR) Salmonella Typhimurium, largely spread through poultry products, increasingly resists antibiotic treatment. We evaluated dietary pomegranate peel extract-loaded nano-emulsions (PomNEs) on performance, immunity, antioxidant capacity, and S. Typhimurium resistance in laying hens. A total of 250 15-week-old Hy-Line Brown hens received a basal diet or diets supplemented with PomNEs at 0.3, 0.6, or 1.2 g/kg, and then were challenged with S. Typhimurium at 34 weeks. Hens fed PomNEs, especially PomNEs1.2, showed improved egg production and feed efficiency before challenge and restored normal output afterward. Supplementation reduced Salmonella colonization in the ovaries, liver, and eggs, with considerable reduction in eggs and ovaries at the highest dose by ten weeks, and down-regulated bacterial virulence genes (hilA, invA). Immune function improved through higher phagocytic activity, intracellular killing, and lysozyme levels, lower nitric oxide, up-regulated IgA and β-defensins (AvBD6, AvBD12), and suppressed pro-inflammatory cytokines and chemokines. Intestinal and ovarian redox balance improved via reduced COX2 and elevated GPX-1, HO-1, NQO1, SOD-1, and CAT expression. These benefits support PomNEs as promising natural feed additives to enhance laying performance, antioxidant defense, and immunity against salmonellosis, offering a sustainable strategy for supporting disease control in poultry production.

Veterinary Sciences doi: 10.3390/vetsci13080774

Authors: Shi’ao Bian Zongzhe Yue Long Zhou Jun Ai Diangang Han Munib Ullah Zhidong Zhang Yanmin Li

Timely detection of specific antibodies against foot-and-mouth disease virus (FMDV) is critical to effective prevention and control of the disease. In this study, we developed a competitive chemiluminescent enzyme immunoassay (cCLEIA) based on recombinant FMDV VP2 protein (FMDV-VP2 cCLEIA) for simultaneous detection of antibodies against FMDV serotypes O and A in serum samples. Using a cut-off of 35% percentage inhibition (PI), the FMDV-VP2 cCLEIA achieved a relative sensitivity of 101.5% and a relative specificity of 102% compared with commercial FMDV serotypes O and A liquid-phase blocking ELISA (LPBE) kits. The assay showed high specificity for detection of antibodies against FMDV serotypes A and O, with no cross-reactivity observed against positive sera for classical swine fever virus, porcine reproductive and respiratory syndrome virus, and Senecavirus A. Both intra-batch and inter-batch coefficients of variation were below 15%, confirming excellent assay reproducibility. The testing of 40 field serum samples yielded a positive detection rate of 95% using the FMDV-VP2 cCLEIA, compared with 92.5% using the FMDV serotypes O and A LPBE kits. In conclusion, the FMDV-VP2 cCLEIA developed is a rapid, sensitive assay for simultaneous detection of antibodies against FMDV serotypes O and A, which may offer a valuable tool for foot-and-mouth disease (FMD) surveillance and control programs.

Veterinary Sciences doi: 10.3390/vetsci13080773

Authors: Chiara Ortello Viviana Manzulli Lorenzo Pace Antonella Cristina Romano Mariateresa Toce Lucia Palazzo Donatella Farina Laura Del Sambro Valeria Rondinone Luigina Serrecchia Domenico Galante

In recent years, the wild boar population has increased in Italy. Wild boar is one of the most problematic species from a sanitary point of view because it represents a dangerous reservoir of pathogens and viruses. The aim of this study was to investigate the circulation of coronaviruses in wild boars, a group of viruses known for their ability to spill over between species. As part of the wild boar population control plan established by the Basilicata region relating to this species, 625 animals were analyzed from May 2024 to January 2026. Nasal swabs were collected at the slaughterhouse in the province of Potenza. Screening was conducted using pan-coronavirus PCR through a nested PCR assay targeting the conserved RNA-dependent RNA polymerase (RdRp) region. Out of all samples analyzed, 1.9% were positive for Porcine Hemagglutinating Encephalomyelitis Virus (PHEV), marking the first identification of this neurotropic coronavirus in wild boars in Italy. The detection of PHEV in free-ranging populations highlights the potential for viral circulation at the wildlife–livestock interface and underlines the importance of integrated surveillance systems to prevent possible interspecies transmission and minimize the risk of spillover to domestic pig herds.

Veterinary Sciences doi: 10.3390/vetsci13080772

Authors: Adrián Alejandro Corrales-Hernández Patricia Roldán-Santiago Héctor Orozco-Gregorio Luis A. de la Cruz-Cruz Ofelia Limón-Morales Juan Manuel Vázquez García Herlinda Bonilla-Jaime

Optimizing farrowing management is crucial in swine production, yet the combined effects of induction protocols and stimulants on the sow remain poorly understood. This study evaluated the effects of farrowing induction using cloprostenol as a single dose (full dose) or split dose, with or without caffeine administration, on the performance, behavior, and hormonal concentrations of primiparous (n = 25) and multiparous (n = 25) sows. Sows were assigned to five treatments: Control (n = 10), PGF2α Full Dose (175 μg) (n = 10), PGF2α Split Dose (87.5 + 87.5 μg) (n = 10), PGF2α Full Dose + Caffeine (420 mg) (n = 10), and PGF2α Split Dose + Caffeine (420 mg) (n = 10). The PGF2α Split Dose and PGF2α Split Dose + Caffeine treatments significantly reduced farrowing duration by up to 100 min in primiparous sows (122.40 ± 82.24 and 136.00 ± 38.33 min, respectively; p = 0.002), and in multiparous sows by up to 76 min (134.80 ± 73.84 and 158.00 ± 44.82 min, respectively; p < 0.05), compared to the Control and PGF2α Full Dose groups. Furthermore, the PGF2α Split Dose induction method lowered the frequency of stress-related behaviors (0.01 and 0.08 for PGF2α SD, and 0.03 and 0.08 for PGF2α SD + Caffeine, for gilts and sows respectively), with sows showing fewer posture changes and maintaining more stable resting positions, particularly the lying lateral posture (0.86 for PGF2α SD and 0.96 for PGF2α SD + Caffeine). Cortisol levels varied by parity, with multiparous sows showing higher prepartum and intrapartum levels, while primiparous sows peaked twelve hours postpartum; estradiol levels remained unaffected by treatments. In conclusion, inducing farrowing with a split-dose prostaglandin protocol is highly efficient, and the addition of caffeine further enhances farrowing performance while significantly improving behavioral welfare by reducing standing postures associated with discomfort in both primiparous and multiparous sows.

Veterinary Sciences doi: 10.3390/vetsci13080771

Authors: Sophia Shanlly Jordan Slessor Wenting Yan Eva D. C. Rosaasen-Uwiera Jessica J. D. Thorlakson Heather L. Bruce Richard R. E. Uwiera

Cushing’s syndrome is a common endocrine disorder in dogs managed with both medical and non-medical treatments; however, the effects of non-medical interventions on survival, perioperative, and clinical outcomes remain uncertain. A comprehensive search was conducted across MEDLINE, Embase, Web of Science, Academic Search Complete, and the Cochrane Library databases. Five studies (n = 248 dogs) met the inclusion criteria. Three studies evaluated laparoscopic and open adrenalectomy in dogs with adrenal-dependent hypercortisolism, whereas two studies evaluated radiotherapy in dogs with pituitary-dependent hypercortisolism. Meta-analysis demonstrated no significant differences in the survival rates between laparoscopic and open adrenalectomy at 12, 24, or 36 months. Fractionated radiotherapy improved survival rates at 12 months, while both fractionated and stereotactic radiotherapy improved survival rates at 24 months. This benefit, however, was not maintained at 36 months. Radiotherapy was also associated with improved neurologic outcomes (OR = 109.17, 95% CI 3.99–2985.49) and longer survival time (MD = 624.00 days, 95% CI 227.93–1020.07). Among dogs undergoing adrenalectomies; laparoscopic adrenalectomy was associated with shorter surgical times (MD = −28.38 min, 95% CI −41.39 to −15.38) and hospitalization (MD = −0.66 days, 95% CI −1.00 to −0.32) compared to open adrenalectomy.

Veterinary Sciences doi: 10.3390/vetsci13080770

Authors: Elshymaa A. Abdelnaby Abdulrhman K. Alhaider Ibrahim A. Emam Hassan S. Al Qarnin

This study aimed to determine the action of a single dose of human chorionic gonadotropin (hCG) on follicular dynamics, CL functionality, ovarian (OV.A) and uterine arteries (MUA) blood flow, hormones (estradiol [E2] and luteinizing hormone [LH]), nitric oxide (NO), and vascular endothelial growth factor (VEGF) levels in Thoroughbred mares. Animals were subjected to ultrasonography scanning, when the dominant follicle (F1) reached 35 mm in diameter, mares were injected with hCG (hCG group; n = 7) at hour 0, while others were injected with saline (control; n = 7). The examination was performed a those points at hours −24 h, −12 h, 0 h, 12 h, 24 h, 36 h, 48 h, 72 h, and 96 h. Blood sampling, ultrasound scanning, hormone assaying, and statistical analysis were performed. F1 was ruptured at 12 h after injection compared to 24 h in the control, with an elevation in its diameter (p ≤ 0.05) and a colored area surrounding F1. The amount of color Doppler area around CL and its diameter were increased (p ≤ 0.05) at 24 and 48 h after injection with an increase (p ≤ 0.05) in the ipsilateral uterine horn colored area from 6 to 36 h after injection. OV.A and MUA Doppler velocity parameters were elevated (p ≤ 0.05) at 24 h. Both NO and VEGF levels were elevated at 24 h after injection, with the elevation (p ≤ 0.05) of E2 occurring only before injection and then declining. hCG and LH levels were increased after injection at 12 and 24 h, respectively, with an interaction (p ≤ 0.05) effect. hCG may speed up and hasten the ovulation process by elevating follicular color amount and CL morphometry via elevating ovarian flow and uterine body total coloration via elevating uterine blood flow (declining Doppler indices) in mares.

Veterinary Sciences doi: 10.3390/vetsci13080769

Authors: Valentina Chisu Rosanna Zobba Laura Giua Cipriano Foxi Piera Bianco Giovanna Chessa Giovanna Masala Alberto Alberti Pierangela Cabras

Tick-borne diseases are an increasing concern in Mediterranean livestock systems, where environmental conditions and extensive grazing favour interactions among hosts, vectors, and pathogens. This study investigated the occurrence and diversity of tick-borne pathogens in small ruminants and their associated ticks in Sardinia (Italy), with the aim of characterizing the occurrence and diversity of tick-borne pathogens in small ruminants and their associated ticks in Sardinia. A cross-sectional survey was conducted on 17 farms in central-eastern Sardinia. Blood samples from 75 animals (57 goats and 18 sheep) and 364 ticks collected from infested hosts were analyzed. Ticks were morphologically identified, and both ticks and blood samples were screened for major bacterial and protozoan pathogens using rt-PCR and conventional PCR assays, followed by sequencing. Among examined hosts, Anaplasma spp. DNA was detected in 28% of goats and 39% of sheep, whereas Theileria ovis was identified in 4% of goats and 78% of sheep. No animals tested positive for Rickettsia spp. or Coxiella spp. Tick analysis revealed a predominance of Rhipicephalus sanguineus and Rhipicephalus bursa, with several tick-borne microorganisms detected, including Rickettsia spp. (26.6%), Coxiella spp. (22.5%), Anaplasma spp. (17.6%), and piroplasms (11.8%). Most Coxiella-positive ticks carried DNA of the Coxiella-like endosymbiont Candidatus Coxiella mudrowiae, while only one tick was positive for Coxiella burnetii. Sequencing confirmed the presence of zoonotic species such as Rickettsia massiliae, Rickettsia sibirica/barbarica group, and Rickettsia hoogstraalii. These findings reveal a complex epidemiological scenario characterized by frequent detection of tick-borne microorganisms in ticks but limited detection in hosts. The presence of zoonotic agents highlights potential public health risks and supports the need for integrated surveillance and control strategies in Mediterranean pastoral systems.

Veterinary Sciences doi: 10.3390/vetsci13080768

Authors: Marko Legler Kristin Heenemann Ingo Gerhauser

Members of the Genus Avipoxvirus have been reported in more than 374 avian species worldwide. Generally, poxviruses are considered to have a limited host range; however, canarypox-like viruses appear to infect a broader host range. In 2024, four juvenile common swifts in an age of 30 to 35 days from three different breeding sites of Lower Saxony, Germany, showed nodular, verrucous and crusted skin lesions of the wings, especially of the dorsal surface of the elbow and forearm region and the skin between the outer primary flight feathers. The skin on the head or the feet as well as the mucous membrane of the oral cavity was not affected. Two swifts of these cases had to be euthanized due to pox-related alterations of feather follicles of the primary flight feathers and associated feather growth disorders and loos. The other two swifts were released into the wild after recovery and successful hand-rearing. The diagnosis of a poxvirus infection is based in our cases on a combination of clinical examination, histology, transmission electron microscopy, and molecular biological investigations. The histological examination of skin lesions displayed severe heterophilic and histiocytic dermatitis with severe epithelial hyperplasia, mild orthokeratotic hyperkeratosis and cytoplasmic eosinophilic inclusion bodies (Bollinger’s inclusion bodies) within frequently swollen keratinocytes (ballooning degeneration). Using transmission electron microscopy, virions showing typical brick-shaped poxvirus morphology were visualized. Avipoxvirus-specific DNA was detected by PCR. The sequence analysis of the 176 bp amplicon revealed 100% identity to a canarypox virus and a shearwaterpox virus 2 strain. Herpesvirus- or papillomavirus-specific DNA was not detected by PCR. To our knowledge, this is the first report of an avipoxvirus infection in a swift species. Notably, in all swift cases, the disease was limited to the skin of the wings. The origin of the virus and its transmission to swifts are unknown. However, transmission from other wild birds via biting insects appears to be possible, as has been described for canarypox-like viruses.

Veterinary Sciences doi: 10.3390/vetsci13080767

Authors: Camilla Schiesari Luigi Tucci Arianna Costa Laura Ventura Giovanna Bertolini Elisa Bortolami

This retrospective case series aims to assess the effects of an anaesthetic protocol including low-dose dexmedetomidine and butorphanol in dogs referred for cardiac tamponade undergoing ECG-gated CCT with contrast medium administration. Fifteen dogs (7 males, 8 females; ASA III/IV, body weight 25 ± 17.5 kg and age 8.6 ± 3.2 years) of different breeds with mild (n = 10/15) and moderate (n = 5/15) pericardial effusion were included. Pericardiocentesis was performed before anaesthesia. Dogs were premedicated with intravenous (IV) dexmedetomidine (0.5–1 μg kg−1) and butorphanol (0.1–0.3 mg kg−1). General anaesthesia was induced with propofol to effect (2–4.5 mg kg−1) IV and maintained with isoflurane in medical air and oxygen. During anaesthesia, heart rate and mean arterial pressure ranged from 37–141 beats min−1 and 58–117 mmHg respectively. No arrhythmias were recorded; mild hypotension occurred in two patients under general anaesthesia. No mortality was recorded within 48 h. Image quality was rated as excellent or good in most examinations, with moderate quality reported in two cases. A protocol including low-dose dexmedetomidine and butorphanol was feasible and was associated with acceptable recorded cardiovascular variables in this case series.

Veterinary Sciences doi: 10.3390/vetsci13080766

Authors: Muhammad Shahzaib Ashfaq Mohamed Tharwat Sohail Ahmed Aftab Shaukat Nourhan Nassar Samavia Abid Muhammad Ahmad Abid Fahad A. Alshanbari

Dairy production currently faces three converging challenges: the escalation of antimicrobial resistance (AMR), rising global food demand, and stricter regulatory requirements for reducing enteric methane emissions. This review evaluates probiotics, prebiotics, fecal microbiota transplantation (FMT), metagenomic tools, and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based synthetic biology for dairy cow microbiome engineering, applying a Technology Readiness Level (TRL) 1–9 framework to assess the translational maturity of each strategy. The quantitative ranges below are from individual controlled or field studies unless indicated otherwise, and they represent the variation from study to study in different breeds, feeds, and stages of lactation, as well as in management systems. A systematic literature search was conducted across five major databases for the period 2020–2026. Applying the TRL framework revealed that conventional probiotics have reached field-ready maturity (TRL 7–8), boosting milk yield by 0.5–1.5 kg/d and lowering somatic cell counts by 20–40%. Calf gut maturation was found to be two to three weeks faster when FMT was used (TRL 5–6). Controlled conditions (TRL 2–3) showed a 10–20% reduction in methane emissions using engineered rumen bacteria (CRISPR). Intervention failures primarily stem from host-microbiome misalignment rather than microbial product design. The key translational gap is shifting from uniform herd-level to precision-guided individualized dosing. Standardized data infrastructure, regulatory frameworks for engineered biologics, and integration with precision livestock farming platforms are required to reduce antibiotic use and lower methane emissions within a One Health framework.

Veterinary Sciences doi: 10.3390/vetsci13080765

Authors: Xiaoyue Yuan Xizhu Zhu Chuyang Zhu Yunhan Wang Chenhui Li Minghua Yu Fanfan Meng Ping Hu Haoyu Liu Qu Chen Demin Cai

Fueled by the rapid growth of the global companion animal industry, the development of nutritional additives for improving canine intestinal health has emerged as a prominent research area in pet nutrition. Excessive reactive oxygen species accumulation triggers severe oxidative stress, inducing intestinal inflammation, abnormal epithelial senescence, and tight junction destruction to damage intestinal mucosal integrity. Omega-3 polyunsaturated fatty acids (ω-3 PUFAs) have proven antioxidative and anti-inflammatory potency, yet divergent functions of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) on canine ileal epithelial cells (cIECs) remain unclear. Here, an H2O2-stimulated oxidative injury model was built to compare their protective capacity. Under all equivalent gradient concentrations ranging from 2.5 to 10 μM in this experiment, Equal-dose DHA failed to alleviate cell damage, while EPA effectively rescued impaired cIEC viability. Biochemical tests showed EPA lowered malondialdehyde and boosted key antioxidant enzyme activities. Transcriptomics confirmed EPA suppressed overactivated TNF-α/IFN-γ inflammatory and senescence pathways. Western blot verified EPA downregulated p-Chk1, p53, p57, and p73 and restored telomere-protective and tight junction proteins. In summary, EPA protects cIECs via antioxidation, anti-inflammation and anti-senescence, supporting its development as a promising functional feed additive for canine intestinal care.

Veterinary Sciences doi: 10.3390/vetsci13080764

Authors: Huaming Xi Jiacun Liu Jing Wang Li Zhong Yigang Xu Yuan Li

The intestinal barrier is a key interface maintaining host–microbial segregation and systemic homeostasis. A broad range of viruses, including enteric, respiratory, and systemic pathogens, can disrupt this barrier through effects on epithelial integrity, vesicular transport, immune responses, and microbial ecology. Whether these diverse insults converge on shared regulatory nodes or act through distinct virus-specific pathways that ultimately result in barrier failure remains unclear. Building on this premise, this review systematically delineates the molecular and cellular mechanisms underlying virus-induced disruption of the intestinal barrier. Viral infection disrupts epithelial integrity through multiple converging processes, including disassembly of tight junction architecture, activation of programmed cell death pathways, degradation of the mucus layer, impaired regeneration driven by intestinal stem cells, and dysregulation of transcellular transport. These processes are interconnected and collectively drive epithelial dysfunction and barrier breakdown. Beyond epithelial damage, we further highlight the pivotal contribution of host immune responses to barrier breakdown. Viral infection induces dysregulated cytokine production and aberrant immune activation, which amplify epithelial damage and further increase barrier permeability. In parallel, increasing evidence supports a bidirectional interaction between viral infection and gut microbiota dysbiosis, in which each process reinforces the other to accelerate barrier disruption and disease progression. We also discuss emerging therapeutic strategies aimed at restoring intestinal homeostasis, including antiviral therapies, host-targeted interventions, and microbiota modulation. Despite recent progress, key questions remain, particularly regarding mechanisms of failed barrier repair after viral clearance and the multilayered regulatory networks linking viruses, immunity, and the microbiota. Together, this review provides a framework for understanding virus-induced intestinal barrier dysfunction and identifies potential therapeutic nodes for intervention.

Veterinary Sciences doi: 10.3390/vetsci13080762

Authors: Md Hashibur Rahman Hyuncheol Jeon Haham Kim Seunghyung Lee

Fishmeal reduction is central to sustainable aquafeed development, but low-fishmeal diets may not reproduce all the nutritional and functional properties of fishmeal after lysine and methionine correction. This PRISMA 2020-based systematic review synthesized peer-reviewed feeding trials evaluating nutritional-balancing strategies beyond these two amino acids in low-fishmeal, fishmeal-free, or high-alternative-protein diets. Alternative protein ingredients were treated as dietary matrices rather than interventions, and studies evaluating ingredient replacement alone were excluded. Twenty-three studies were included. Study-level effect sizes were estimable for 21 studies, providing 24 representative performance-related comparisons. Eight comparisons had 95% confidence intervals excluding zero in favor of the intervention, whereas 16 crossed the null; estimates were not pooled because of substantial heterogeneity in species, diet formulation, intervention, dose, and outcome definition. The evidence covered essential and functional amino acids, taurine and related nitrogenous compounds, palatability enhancers, enzymes, phosphorus and mineral balance, antioxidant support, lipid-associated nutrients, and gut-targeted strategies. Phytase provides the clearest mechanism through which phosphorus utilization can be improved in phytate-rich or phosphorus-limited diets. Taurine responses were generally favorable but imprecise in the four marine finfish species. Protein hydrolysates, fermented ingredients, yeast products, probiotics, hydroxyproline, and other additives produce context-dependent benefits for performance, nutrient utilization, intestinal conditions, immunity, stress resistance, or product quality. Across strategies, efficacy depended mainly on matching the intervention to the nutritional or functional limitation created by the complete diet. Nutritional equivalence should therefore be evaluated via integrated production, mechanism-specific, health, product-quality, environmental, and economic outcomes rather than the fishmeal-replacement percentage alone.

Veterinary Sciences doi: 10.3390/vetsci13080763

Authors: Rui Tang Jianfei Zhao Yong Yang Jun Yan Shanchuan Cao Zhengqun Liu Jingbo Liu

This study evaluated the effects of low-level replacement of corn with dried bamboo shoot powder (DBSP) on growth performance, nutrient digestibility, serum biochemical, immune, inflammatory, and antioxidant indices, colonic fermentation, microbiota, carcass traits, and meat quality in growing–finishing pigs. A total of 128 healthy pigs (Landrace × Yorkshire × Duroc; initial body weight, 82.90 ± 1.71 kg) were randomly assigned to four dietary treatments: a basal diet (CON) and diets in which 1% (DBS1), 2% (DBS2), or 3% (DBS3) of corn was replaced with DBSP. The experiment included a 5-day adaptation period and a 35-day experimental period. DBSP replacement did not affect growth performance, serum biochemical, immune, inflammatory, or antioxidant indices, or meat quality (p > 0.05). Crude protein apparent total tract digestibility increased linearly with DBSP level and was higher in DBS3 than in CON (p < 0.05). Colonic acetate concentration increased linearly, and total short-chain fatty acid concentration was affected by dietary treatment. Carcass weight and dressing percentage showed linear responses to increasing DBSP replacement levels. Overall, replacing corn with up to 3% DBSP did not significantly impair growth performance or meat quality and may influence crude protein apparent total tract digestibility, colonic fermentation, and selected carcass traits.

Veterinary Sciences doi: 10.3390/vetsci13080761

Authors: Alma García-Ramos Omar García-Pérez Angélica Domínguez-de-Barros Malena Gajate-Arenas Candela Sirvent-Blanco José E. Piñero Jacob Lorenzo-Morales Elizabeth Córdoba-Lanús

Rickettsia is an obligate intracellular genus of bacteria responsible for rickettsioses, a group of emerging vector-borne diseases. Their diagnosis remains challenging because of non-specific clinical manifestations, serological cross-reactivity, and low bacterial loads in clinical samples. In this study, a novel droplet digital polymerase chain reaction (ddPCR) assay was developed for the molecular detection of Rickettsia spp., and its performance was evaluated using clinical blood samples collected from dogs and cats in the Canary Islands, Spain. Primers and probes targeting the 16S ribosomal RNA (rRNA) were designed and optimized for ddPCR amplification. Analytical performance was assessed by optimising primer and probe concentrations, annealing temperature, specificity, and the limit of detection (LOD). Comparative analysis between quantitative PCR (qPCR) and ddPCR was also performed. The optimized assay employed primer and probe concentrations of 900 nM and 250 nM, respectively, with an annealing temperature of 56 °C. The assay achieved an analytical detection limit of 0.064 ng of clinical sample DNA, although sporadic positive droplets were still detectable at lower concentrations. Strong inverse correlations were observed between qPCR cycle threshold (CT) values and ddPCR absolute quantification for both plasmid DNA (r = −0.997) and clinical sample DNA (r = −0.983). A total of 170 blood samples collected from domestic animals in the Canary Islands, with clinical suspicion of rickettsiosis, were analysed. Six canine samples (3.5%) tested positive for Rickettsia spp. The ddPCR assay developed in this study showed promising analytical performance for the molecular detection of Rickettsia spp. DNA and has the potential to become a valuable molecular tool for veterinary diagnostics and epidemiological investigations within a One Health framework. Further analytical and clinical validation studies are required before its implementation as a routine diagnostic assay.

Veterinary Sciences doi: 10.3390/vetsci13080760

Authors: Agnieszka Noszczyk-Nowak Piotr Frydrychowski Justyn Gach Zuzanna Wojtczak Alicja Cepiel-Kośmieja Artur Fuglewicz Krzysztof Nowak

Radiofrequency catheter ablation is a minimally invasive electrophysiological procedure that utilizes radiofrequency energy to induce targeted thermal injury within arrhythmogenic myocardial tissue, thereby eliminating the substrate responsible for rapid and irregular cardiac rhythm disturbances. Radiofrequency ablation is a method widely used in human medicine for the permanent treatment of arrhythmias. In veterinary medicine, there are only a few centers worldwide that use this treatment method in dogs. This article presents the experiences of a Polish center in treating supraventricular and ventricular arrhythmias using this method. The first electrophysiological study and ablation were performed using an analog electrophysiological system; subsequent procedures were performed with the aid of a computerized system, and the last three procedures were performed using a 3D electroanatomical system. A total of 14 radiofrequency ablation procedures were performed on dogs weighing 10 to 33 kg, aged 9 months to 9 years, of various sexes and breeds. Fifty percent of the dogs that underwent ablation had both clinical and echocardiographic signs of tachycardia-induced cardiomyopathy. Three dogs required a re-ablation procedure. Complete elimination of clinically significant arrhythmias was achieved in 13 of 14 dogs, while one dog showed partial procedural success. No significant or permanent complications were observed in any of the dogs. Radiofrequency ablation procedures for arrhythmias are effective and safe.

Veterinary Sciences doi: 10.3390/vetsci13080759

Authors: Li-Lian Chao Chien-Ming Shih

Anaplasma platys was recognized as a zoonotic pathogen for canine and human infections. However, there is no confirmed evidence demonstrating the existence of A. platys within hard ticks (Rhipicephalus sanguineus sensu lato) infesting dogs in Southern Taiwan. We conducted a field survey for the detection of Anaplasma infection by screening the R. sanguineus s. l. ticks collected from dogs. Molecular detection was performed via a nested-PCR assay targeting the 16S rRNA gene of Anaplasma. Phylogenetic analyses were performed via neighbor-joining against maximum parsimony methods. The genetic relatedness was analyzed via pairwise distance calculation, using the Kimura 2-parameter model. The A. platys infection was detected in various life stages of R. sanguineus s. l. ticks, with an infection rate of 0.8%, 1.0% and 1.4% in nymphs, males and females, respectively. Monthly prevalence was observed from July to October. The genetic identity of detected Anaplasma strains from Taiwan was phylogenetically affiliated to the A. platys species with a 100% sequence homology verified from different countries. This study reveals the first molecular evidence of a zoonotic pathogen, A. platys, detected in R. sanguineus s. l. ticks from Southern Taiwan. Further investigation focused on the vectorial capacity of R. sanguineus s. l. ticks for transmitting A. platys will be beneficial to reveal the public health importance regarding the potential risk for canine and human infection in Southern Taiwan.

Veterinary Sciences doi: 10.3390/vetsci13080758

Authors: Ya Wang Zhengyi Shen Ling Lv Zhiguo Li Chen Yu Renwu Zhou Yunhao Su Youtian Deng Junliang Deng Huidan Deng

This study aimed to explore the protective effects of different concentration combinations of four traditional Chinese medicine (TCM) monomers (Eleutheroside E, Anemoside B4, Forsythoside A, and Esculin) on porcine intestinal epithelial cell (IPEC-J2) damage induced by soybean 7S globulin, providing preliminary in vitro evidence for future studies on dietary-allergen-induced intestinal injury in piglets. An L9 (34) orthogonal experimental design was implemented to identify the optimized ratio of these monomers. IPEC-J2 cells were co-cultured with 5 mg/mL 7S globulin and the selected combinations for 24 h. Cell viability was determined via Cell Counting Kit-8 (CCK-8) assay, while cytokine secretion, oxidative status, and mechanical barrier markers were assessed using enzyme-linked immunosorbent assay (ELISA) and RT-qPCR. The optimal combination was determined to consist of Eleutheroside E at 50 mg/L, Anemoside B4 at 25 mg/L, Forsythoside A at 80 mg/L, and Esculin at 20 mg/L. This formulation significantly alleviated inflammatory damage, upregulated tight-junction-related mRNA expression, and reversed the decrease in cellular viability caused by 7S globulin. Furthermore, Western blot and transcript analyses suggested that this combination exerted its protective effects by modulating the nuclear factor-kappa B/mitogen-activated protein kinase (NF-κB/MAPK) pathway-related markers, reducing the expression of p38 MAPK, c-Jun N-terminal kinase (JNK), NF-κB p65, and inducible nitric oxide synthase (iNOS). In conclusion, this optimized combination of TCM monomers successfully alleviates both inflammatory injury and oxidative stress in IPEC-J2 cells in vitro and holds potential as a promising dietary additive candidate for mitigating dietary-allergen-induced enteropathy in piglets.

Veterinary Sciences doi: 10.3390/vetsci13080757

Authors: Baorui Liu Guoqiang Cui Haiyan Xiao Zhiqiang Peng Jianguo Tan Xianren Jiang Kun Meng

This study aimed to assess the effects of replacing 20% of the standard diet with fermented forage-based mixed feed (FF) on finishing pigs. The Pennisetum No. 1 forage powder, feed ingredients, and composite microbial inoculum were mixed, and the mixture was incubated for 7 days at room temperature, maintained above 25 °C. A total of 176 finishing pigs were randomly assigned to two dietary treatments: a basal diet (CON) and an experimental diet with 20% of the basal diet substituted by FF (20% FF). Each treatment consisted of four replicates, with each replicate including 22 pigs. Throughout the 56-day trial, the findings showed that there were no notable differences (p > 0.05) in growth performance, apparent nutrient digestibility, carcass characteristics, and muscle fatty acid profiles between the control group and the 20% FF group. However, the dietary inclusion of 20% FF significantly enhanced glutathione peroxidase (GSH-Px) activity by 46.59% and decreased the malondialdehyde (MDA) concentration by 22.79% in the Longissimus thoracis et lumborum muscle (p < 0.05), suggesting enhanced muscular antioxidant capacity. In summary, substituting 20% of the basal diet with FF represents a feasible approach that may improve the antioxidant capacity of pork while sustaining both growth performance and carcass characteristics in pigs. This research establishes a scientific foundation for utilizing FF as a practical alternative feed resource in finishing pig production.

Veterinary Sciences doi: 10.3390/vetsci13080756

Authors: Md Hashibur Rahman Hyuncheol Jeon Haham Kim Seunghyung Lee

Thermal variability associated with climate change is an increasing constraint for finfish aquaculture because water temperature directly affects feed intake, nutrient utilization, endocrine stress responses, oxidative balance, immunity, intestinal integrity and survival. The literature search was conducted without a lower publication-year restriction through 5 July 2026. Database and citation-linked searches identified 386 candidate records, of which 274 remained after duplicate removal; 71 full-text articles were assessed; and 28 peer-reviewed primary feeding studies with publicly available final versions of record were retained for qualitative evidence mapping. The eligible studies were published between 2012 and 2026, with 2012 representing the earliest eligible publication rather than a prespecified starting year. Functional aquafeeds have therefore emerged as promising nutritional tools for supporting physiological resilience under heat, cold and sub-optimal temperature conditions. This review developed a PRISMA-guided systematic evidence map of dietary functional-aquafeed strategies evaluated for thermal-stress tolerance, immune regulation, antioxidant defense and metabolic homeostasis in cultured finfish. The main synthesis emphasizes studies with direct dietary thermal-stress interventions and extractable mechanistic outcomes, including sodium butyrate, organic selenium, selenium nanoparticles with riboflavin, astaxanthin, Bacillus spp., propolis, bay laurel essential oil, Spirulina-based strategies and essential oils. Overall, the available evidence suggests that selected functional aquafeeds may support thermal-stress resilience through coordinated improvements in antioxidant capacity, reduced oxidative damage, modulation of cortisol/glucose and heat-shock responses, enhancement of innate and mucosal immunity, improved intestinal or tissue condition and stabilization of metabolic indicators. The most biologically coherent translational patterns were observed when dietary interventions improved multiple response domains simultaneously rather than a single biomarker. However, such cross-domain improvement was interpreted as mechanistic and translational support rather than as an independent indicator of evidence confidence, which depended primarily on independent replication, consistency of response direction and methodological quality. Nevertheless, the current evidence remains uneven across species, stress models, life stages and outcome domains, with limited data for marine carnivores, repeated temperature fluctuation, post-stress recovery, disease challenge and multiomics integration. Therefore, the findings should be interpreted as a structured evidence map rather than universal formulation guidance. Future studies should use standardized thermal-challenge protocols, dose–response designs, clear tank-level experimental-unit reporting and integrated biomarker panels to support practical development of climate-resilient functional aquafeeds.

Veterinary Sciences doi: 10.3390/vetsci13080755

Authors: Alexandr Ostrovskii Anara Ryskeldina Viktoriya Lucay Ayan Dauletov Rashid Karmaliyev Kassym Mukanov Alexandr Shevtsov Assel Akhmetova

Bovine babesiosis is a tick-borne disease of major veterinary and economic importance, yet molecular epidemiological data from Kazakhstan remain limited. This study investigated the prevalence, distribution, and species composition of Babesia spp. in cattle across all 17 administrative regions of Kazakhstan. A total of 6455 cattle blood samples from 171 villages were screened by nested PCR targeting the cytochrome b (cytb) gene, and positive samples were characterized by sequencing and phylogenetic analysis. The overall positive rate for Babesia spp. were 3.87% (95% CI: 3.41–4.37%). Of 250 PCR-positive samples, 215 yielded high-quality sequences and were assigned to four species: Babesia major (31.2%), B. occultans (29.6%), B. bigemina (18.0%), and B. bovis (7.2%). Marked regional heterogeneity was observed, with B. major and B. occultans predominating across most regions, whereas B. bigemina was dominant in Atyrau. Turkistan was the only region where all four species co-circulated. First molecular evidence of B. bovis and B. bigemina was obtained for Atyrau, Akmola, and West Kazakhstan regions. These findings expand current knowledge of bovine babesiosis in Kazakhstan and reveal a species composition dominated by B. major and B. occultans, contrasting with the B. bovis/B. bigemina-dominated patterns commonly reported elsewhere. The detection of pathogenic Babesia species in previously unsurveyed regions highlights the need for continued molecular surveillance.

Veterinary Sciences doi: 10.3390/vetsci13080754

Authors: Stefano Sartore Riccardo Moretti Stefania Chessa Paola Sacchi

Aggression is a behavioral phenomenon observed not only in wild animals but also in companion animals, including the domestic cat. Although feline aggression represents a common welfare and public health concern, its genetic and environmental causes remain not fully understood. The aim of this systematic review was to identify and synthesize the current evidence regarding the genetic and environmental factors associated with aggressive behavior in domestic cats. Original articles published in English between 2005 and 2025 were retrieved from Scopus and PubMed using predefined search strategies. After a two-stage screening and eligibility assessment, 25 studies retrieved though the database searches met the inclusion criteria. One additional eligible study was identified through reference screening, resulting in a total of 26 included studies. No formal risk-of-bias assessment was performed. Findings were synthesized narratively because of the substantial heterogeneity in study design, behavioral assessment methods, and outcome measures. The available evidence indicates that feline aggression is influenced by a complex interaction between genetic predisposition and environmental factors. Associations have been reported with polymorphisms in genes such as OXTR and AR, chromosomal abnormalities, breed, sex, coat color, early-life experiences, owner management, and environmental conditions. Considerable variability in behavioral assessment methods and the absence of a universally accepted classification of feline aggression currently limit direct comparison across studies. Overall, feline aggression should be regarded as a multifactorial trait whose accurate interpretation requires consideration of both inherited and environmental influences.

Veterinary Sciences doi: 10.3390/vetsci13080753

Authors: Qiuyang Sun Yufang Feng Fangwei Dai Qiang Gao Xueqing Zhang Shanshan Yao Rui Fu Chunnan Liang Jin Xing

A multiplex liquid array assay based on Luminex xTAG technology was developed for the first time for the simultaneous detection of five respiratory bacterial pathogens in Specific Pathogen-Free (SPF) chickens—Mycoplasma gallisepticum (MG), Mycoplasma synoviae (MS), Pasteurella multocida (Pm), Avibacterium paragallinarum (Apg), and Mycobacterium avium (Mav)—filling the gap that these five pathogens could not be simultaneously detected previously. Specific primers were designed targeting conserved genes of each pathogen, with a TAG sequence attached to the 5′ end of each forward primer and biotin to the 5′ end of each reverse primer. After multiplex PCR amplification, the products were hybridized with MagPlex-TAG microspheres and analyzed using the Luminex system. The method was validated for sensitivity, specificity, and repeatability, and tested with simulated positive samples (a mixture of genomic DNA from the five target pathogens) and SPF chicken samples. The LOD was 10 copies/μL for Pm and 100 copies/μL for the other four pathogens. No cross-reactivity was observed with 18 non-target bacterial species. Intra- and Inter-assay coefficients of variation (CV) ranged from 0.8% to 12.2% and 2.3% to 14.4%, respectively, both below 15%. All simulated mixed positive samples yielded results consistent with expectations, and none of the 114 nasopharyngeal swab samples from SPF chickens tested positive for any of the five targets. The developed assay offers high throughput, high sensitivity, high specificity, and good repeatability, providing a robust tool for routine quality control in SPF chickens.

Veterinary Sciences doi: 10.3390/vetsci13080752

Authors: Joanna Klećkowska-Nawrot Aleksander Chrószcz Abit Aktaş Karolina Barszcz Wojciech Paszta Karolina Goździewska-Harłajczuk Dominik Poradowski

The orbital glands are part of the ocular surface protective system, but their morphology and secretory profiles remain poorly documented in small wild bovids. This preliminary descriptive study examined nine young animals: four Kirk’s dik-diks (Madoqua kirkii) and five red forest duikers (Cephalophorus natalensis). All animals underwent gross anatomical examination, and bilateral Harderian glands (HGs), lacrimal glands (LGs), and superficial glands of the third eyelid (SGTE) were examined histologically, histometrically, and histochemically. No inferential statistical comparisons between species were performed. In both groups, the HG was located medioventrally, the LG dorsolaterally, and the SGTE medially near the third eyelid. The glands showed compound tubuloacinar or predominantly acinar organization. Descriptive variation involved capsule thickness, septal development, lobular arrangement, secretory epithelial appearance, and plasma cell and lymphocyte distribution. Histochemical reactions demonstrated mixed secretory profiles containing neutral glycoconjugates and acidic mucosubstances consistent with sulfated and/or carboxylated components. Their relative proportions varied among glands and between groups. These findings provide preliminary anatomical and microscopic reference data for rarely examined wild bovids. Because of the small sample sizes, the observations should not be interpreted as definitive species-level differences.

Veterinary Sciences doi: 10.3390/vetsci13080751

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

Brucellosis remains among the most widespread global zoonoses, yet its genetic architecture—spanning host susceptibility and pathogen virulence—is increasingly being investigated. Advances in whole-genome sequencing, transposon mutagenesis, CRISPR screening, and machine learning-driven transcriptomics have accelerated marker discovery on both sides of the host–pathogen interface, although most signatures remain at discovery or early-validation stages. In hosts, polymorphisms in innate immune receptors (TLR2, TLR5, VDR), cytokine regulators (TNF, CTLA4), and novel candidates such as ASAP1 are linked to differential susceptibility in humans and livestock. In Brucella, functional genomics shows that virulence relies on interconnected regulatory circuits that control the type IV secretion system, lipopolysaccharide biosynthesis, and stress responses, with gene essentiality shifting according to the host immune context. Antimicrobial resistance may involve unconventional mechanisms, including metabolic and regulatory adaptation, that are not fully explained by classical resistance-gene paradigms and may implicate metabolic reprogramming and ribosomal modulation. This review synthesizes these converging lines of evidence, highlights key gaps—chiefly the scarcity of functional validation for host associations and limited multi-omics integration—and proposes a roadmap for translating genetic markers toward future diagnostic refinement and therapeutic target discovery, urging a shift from descriptive cataloging toward mechanistic and clinical validation.

Veterinary Sciences doi: 10.3390/vetsci13080750

Authors: Silva Costarelli Mario D’Incau Deborah Cruciani Carmen Panzieri Alessandro Fiorucci Sergio Scorcelletti Alessandro Nunzi Maria Grazia Brancaleoni Luca Latini Antonella Catalano

Leptospirosis is a widespread zoonotic disease of major concern for both veterinary and public health. In horses, it is associated with equine recurrent uveitis, a painful condition that may lead to blindness and affects approximately 10% of the global horse population. In May 2025, following the occurrence of three uveitis cases in a riding stable in Central Italy, 94 equine blood samples were collected and tested using the microscopic agglutination test (MAT) for Leptospira spp. Antibody titres against serogroup Icterohaemorrhagiae were detected in 68 horses (72.4%). An on-site investigation was conducted, and control measures were implemented, including movement restrictions, clinical monitoring of horses, and antibiotic treatment. Follow-up serological testing showed no significant variation in antibody titres, no additional clinical cases were observed in animals, and no symptoms were reported among individuals exposed to the facility. These findings suggest widespread exposure to Leptospira spp. without evidence of ongoing transmission or public health impact. This study highlights the diagnostic and management challenges of equine leptospirosis, together with the importance of surveillance within a One Health framework.

Veterinary Sciences doi: 10.3390/vetsci13080748

Authors: Yuxue Yang Tong Guo Haoran Deng Xiaoyi Li Fuhao Chen Liyuan Huang Shan Chen Jiarui He Jiangwei Xiang Haocheng Huang Hongchuan Deng Kun Zhang Zhijun Zhong Ziyao Zhou Guangneng Peng Dechun Chen Xu Song Haifeng Liu

Background: Microplastics (MPs) and cadmium (Cd) are widespread environmental pollutants posing significant health risks, but their combined hepatotoxic effects and underlying mechanisms remain poorly understood. Methods: Using in vivo and in vitro co-exposure models, we systematically investigated the impact of MPs on Cd-induced hepatotoxicity. The study assessed hepatic injury, oxidative stress, and inflammatory responses through transcriptomic analysis, histopathological examination, immunofluorescence, and quantitative real-time PCR. Pharmacological inhibition of endoplasmic reticulum stress with 4-phenylbutyric acid and selective blockade of IRE1α with MKC3946 were employed to dissect the signaling pathway. Results: Co-exposure to MPs and Cd significantly aggravated Cd-induced hepatic pathological damage, inflammation, and oxidative stress. Transcriptomic profiling revealed marked activation of the endoplasmic reticulum stress pathway and upregulation of pyroptosis-associated genes. Mechanistically, endoplasmic reticulum stress triggered pyroptosis via the IRE1α/TXNIP/NLRP3 signaling axis. Notably, inhibition of endoplasmic reticulum stress with 4-phenylbutyric acid, or selective blockade of IRE1α with MKC3946, effectively attenuated TXNIP/NLRP3 activation and the downstream pyroptotic response. Conclusions: MPs intensify Cd-induced hepatotoxicity by activating the IRE1α/TXNIP/NLRP3 pathway, leading to endoplasmic reticulum stress-driven pyroptosis. These findings provide a mechanistic framework for understanding the combined toxicity of microplastics and heavy metals, with important implications for environmental health risk assessment in animals and humans.

Veterinary Sciences doi: 10.3390/vetsci13080749

Authors: Hyungsan Seo Beom-Joon Park Yookyeong Lee Hyung-Seok Seo Hwi-Yool Kim

Functional pheochromocytoma is an uncommon canine adrenal tumor associated with substantial surgical and anesthetic challenges. This report describes two dogs with suspected functional pheochromocytoma that exhibited distinct causes of surgical complexity. Both underwent endocrine evaluation, contrast-enhanced CT, adrenalectomy, and histopathology. In Case 1, severe perivascular adhesion resulted in an iatrogenic caudal vena cava laceration despite the absence of definitive vascular invasion, requiring primary venorrhaphy. In Case 2, marked tumor-induced vascular distortion obscured critical anatomical landmarks, necessitating staged surgical management. Three-dimensional vascular reconstruction facilitated refinement of the surgical approach, while individualized anesthetic management enabled successful surgery despite marked intraoperative hemodynamic fluctuations. Histopathology confirmed pheochromocytoma in both dogs, including malignant pheochromocytoma in Case 2. Case 1 developed chronic kidney disease during long-term follow-up, whereas no clinical evidence of local recurrence or distant metastasis was observed during the available follow-up. These cases demonstrate that surgical complexity may arise from severe perivascular adhesion and tumor-induced vascular distortion, even without planned vascular lumen entry or advanced vascular procedures. Patient-specific anatomical assessment, individualized perioperative planning, and selective 3D vascular reconstruction may improve surgical safety in complex adrenal tumors.

Veterinary Sciences doi: 10.3390/vetsci13080746

Authors: Khondoker Mohammad Iftekhar Zahan Wissanuwat Chimnoi Roshni Jahan Nur Antara Chanya Kengradomkij Narisorn Pilean Ketsarin Kamyingkird Tawin Inpankaew

Gastrointestinal (GI) parasitic infections are still a major concern for cattle health and productivity in tropical and subtropical regions. However, epidemiological data on GI parasitism in cattle from western Thailand remain limited, particularly in border areas with frequent animal movement. This study focuses on establishing the prevalence of gastrointestinal parasitic infections and identifies its related risk factors among the cattle in Kanchanaburi province, western Thailand. Cross-sectional study was performed from December 2022 to January 2023 involving 153 cattle from the following three study populations: local cattle from Thong Pha Phum district, Sangkhlaburi district, and cross-border cattle imported from Myanmar. Fecal samples were examined using both simple flotation and formalin-ethyl acetate sedimentation techniques, and data on animal and management factors were collected through questionnaire surveys. Overall, 35.9% (55/153) of the cattle had at least one GI parasite species positive. Six parasite taxa were identified, with Strongyle-type nematodes being the most frequently detected (26.1%), followed by other helminth parasites. The prevalence of infection and parasite diversity increased progressively in cattle populations located closer to the Thai–Myanmar border, suggesting that this region may represent a potential hotspot for GI parasite transmission. From a One Health perspective, these findings highlight the potential for some helminths to circulate across animal, human, and environmental interfaces. The presence of 1.3% for both Capillaria spp., Trichuris spp. and 0.7% of Fasciola spp. suggests possible zoonotic transmission pathways, particularly via the fecal-oral route. Although 93.5% of cattle had reportedly received anthelmintic treatment, no apparent protective effect was observed. These findings highlight that the Thai–Myanmar border acts as a hotspot for parasite transmission and the need for improved parasite control strategies in this region, alongside integrated One Health surveillance linking animal, human, and environmental health in this cross-border region.

Veterinary Sciences doi: 10.3390/vetsci13080747

Authors: Krisztián Németh Borbála Mózes Tibor Bartha Boglárka Mária Schilling-Tóth Gergely Jócsák Dávid Sándor Kiss Ágnes Sterczer Marianna Kis István Tóth

Bioactive collagen peptides and collagen hydrolysates are dietary proteins whose degradation products also carry signaling activity. This second part of a two-part narrative review addresses the translational dimensions of their bioactivity, integrating veterinary clinical trials, controlled animal-model studies, and human biomedical data, with evidence strictly stratified by type. The gastrointestinal tract acts not only as the absorption site but as a target organ. In cell culture and rodent models, luminal collagen fragments restore tight-junction integrity, alter the microbiome, and shift the enterohepatic bile acid pool; increased secondary bile acid synthesis is proposed to engage the farnesoid X receptor (FXR) and Takeda G-protein-coupled receptor 5 (TGR5). None of these steps have been demonstrated in veterinary clinical patients. The accompanying GLP-1 and PYY response may reflect direct amino acid stimulation of enteroendocrine cells as much as a bile acid-dependent route, and current data do not separate the two. Veterinary trials show objective kinetic improvement in osteoarthritic dogs and horses. Taurine-responsive dilated cardiomyopathy is a reversible, diet-amenable condition; as the principal bile acid conjugation substrate in carnivores, taurine acts on the same bile acid pool as a mechanistically separate input. Recent data implicate prolyl-hydroxyproline in brown adipogenesis, collagen peptides in hippocampal neurogenesis, and taurine in platelet normalisation. The gut–collagen peptide axis is an evolving model linking dietary collagen to systemic metabolic, endocrine, and immune signaling, with implications for companion-animal, equine, and livestock practice.

Veterinary Sciences doi: 10.3390/vetsci13080745

Authors: Tong Guo Yuxue Yang Fuhao Chen Haoran Deng Hongchuan Deng Xiaoyi Li Zhuohang Wu Aoxuan Jiang Haocheng Huang Guangneng Peng Zhijun Zhong Ziyao Zhou Kun Zhang Dechun Chen Haifeng Liu

Microplastics (MPs) can adsorb and transport heavy metals, but their influence on cadmium (Cd)-induced hepatotoxicity in mammals remains unclear. Forty-eight male Kunming mice were assigned to control, Cd, MP, and Cd + MP groups and exposed by oral gavage for 42 days. Growth performance, liver injury, oxidative stress, and inflammation were assessed, and transcriptomic and metabolomic analyses were integrated with qPCR, mitochondrial DNA (mtDNA) copy number, and ATP measurements. Compared with Cd alone, combined exposure resulted in greater reductions in body weight gain, the liver index, and antioxidant enzyme activities, together with more severe hepatic lesions and higher levels of liver injury markers and inflammatory cytokines. Co-exposure also induced broader transcriptional and metabolic disturbances than Cd alone. Integrated omics analyses converged on the dysregulation of AMPK–FOXO signaling and related energy metabolic processes. qPCR confirmed more pronounced alterations in pathway-related genes after co-exposure, while reductions in mtDNA copy number and ATP content indicated aggravated mitochondrial dysfunction and impaired energy metabolism. Collectively, these results demonstrate that MPs exacerbate Cd-induced liver injury and suggest that disruption of AMPK–FOXO-associated energy metabolism is a key molecular feature underlying the enhanced hepatotoxicity observed under combined exposure.

Veterinary Sciences doi: 10.3390/vetsci13080744

Authors: Zhen Liu Ying Bao Tiantian Gu Li Chen Tao Zeng Lizhi Lu Zhizhong Lin

The current study aims to assess the effects of coated sodium butyrate (CSB) on the weaning transition of White King pigeons. A total of 336 1-month-old White King pigeons were assigned randomly to four groups, including the Control group fed basal health sandand without CSB and test groups fed the basal health sand with 0.1%, 0.2%, and 0.4% CSB, respectively. The results showed that, compared with the Control group, the average daily feed intake (ADFI) was significantly increased in the 0.1% and 0.2% CSB groups (p < 0.05). The 0.1% CSB group exhibited a higher immune organ index (IOI) than the Control group (p < 0.05). The total protein (TP) and albumin (ALB) levels in the 0.1% CSB and 0.4% CSB groups significantly increased compared with the Control group (p < 0.05). In contrast, the 0.2% CSB group exhibited lower serum ALB, TP, and total cholesterol (TC) levels; TNF-α and IL-6 concentrations were reduced compared with the other groups (p < 0.05), and the 0.4% CSB group showed significantly increased serum TNF-α, IL-6, and D-lactic acid levels (p < 0.05). The ratio of villus height to crypt depth (VH/CD) of the jejunum (p < 0.05) was significantly increased in the 0.2% CSB group. In addition, the CD and VH/CD of the ileum were significantly increased in the 0.1% CSB group (p < 0.05). Firmicutes was the predominant phylum across all treatment groups, while Bacteroidota showed a more sensitive response to CSB supplementation. The 0.1% CSB group showed a significant enrichment of Oscillospiraceae and Muribaculaceae, fiber-fermenting, SCFA-producing taxa, Enterobacterales and Klebsiella, suggesting CSB-mediated alterations in luminal oxygen tension that favor the coexistence of facultative and obligate anaerobic taxa. Collectively, these data revealed that CSB supplementation efficiently improves growth performance and intestinal health of weaning pigeons by ameliorating the intestinal environment.

Veterinary Sciences doi: 10.3390/vetsci13080743

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

Mammary gland inflammation, clinically expressed as mastitis in dairy ruminants and lactating women, arises from a self-amplifying loop of pathogen sensing, oxidative stress and innate immune activation, and remains a leading driver of antibiotic use, milk-quality loss and morbidity. Mounting pressure to reduce antimicrobial dependence has refocused attention on flavonoids—structurally diverse plant polyphenols with multi-target bioactivity, derived mainly from in vitro and rodent mastitis models. This review integrates contemporary evidence on the six principal flavonoid subclasses within a unifying molecular framework. Across subclasses, flavonoids converge on shared targets: the TLR4–MyD88–NF-κB axis, MAPK cascades, the Keap1–Nrf2–ARE antioxidant pathway, the NLRP3 inflammasome and tight-junction proteins of the blood–milk barrier. Less canonical mechanisms—m6A epitranscriptomic regulation, ferroptosis suppression, AhR signalling, anti-virulence binding to bacterial enzymes such as IGPD, and gut-microbiota-driven remodelling of the gut–mammary axis—expand the pharmacological landscape. We additionally appraise the subclasses comparatively, identifying flavanones and the flavone baicalin as carrying the strongest translational evidence, and examine the conflicting findings, model limitations, and delivery, residue and regulatory barriers that currently separate mechanistic promise from on-farm application. We outline structure–activity considerations and translational priorities, and position flavonoids as mechanism-rich, antibiotic-sparing candidates for the prevention and adjunctive management of mammary gland inflammation in dairy ruminants.

Veterinary Sciences doi: 10.3390/vetsci13080742

Authors: Xiaoyu Hou Yuefei Yang Ruiping Wei Xiaochen Cui Huanyu Jiang Huiming Ju

To preliminarily elucidate the bidirectional metabolic effects caused by changes in PFKM expression, provide research clues for further investigation of the molecular mechanisms through which PFKM regulates porcine skeletal muscle metabolism and myogenic differentiation, and offer a reference for identifying candidate genes associated with meat production traits and studying gene regulation in pig breeding, this study compared skeletal muscle protein expression profiles between Large White and Bama pigs. Candidate regulatory factors related to muscle growth and energy metabolism were screened, and the effects of altered muscle-type phosphofructokinase (PFKM) expression on metabolic homeostasis and myogenic differentiation in porcine skeletal muscle satellite cells (SMSCs) were preliminarily evaluated. Longissimus dorsi muscle tissues from Large White and Bama pigs were analyzed using iTRAQ-based proteomics. A total of 2040 reliably quantified proteins were identified, of which 51 were relatively upregulated in Large White pigs and 73 were relatively upregulated in Bama pigs. Functional enrichment analysis showed that the differentially expressed proteins were mainly involved in glycolysis, mitochondrial energy metabolism, protein synthesis, and the regulation of muscle fiber structure and function. PFKM was therefore selected as a key candidate differentially expressed protein. Porcine SMSC models comprising a PFKM knockdown group (PFKM-KD), a PFKM overexpression group (PFKM-OE), and a normal control group (PFKM-CON) were subsequently established. Glucose consumption and lactate accumulation in the culture medium, ATP levels, reactive oxygen species (ROS), mitochondrial membrane potential, apoptosis, mitochondrial dynamics-related proteins, and myogenic differentiation markers were then examined. Compared with the PFKM-CON group, the PFKM-OE group showed significantly increased glucose consumption and lactate accumulation, together with significant increases in ROS levels, mitochondrial membrane potential, and apoptosis, whereas ATP levels were significantly reduced. In the PFKM-KD group, glucose consumption and lactate accumulation were significantly decreased, accompanied by reductions in mitochondrial membrane potential, ROS, ATP levels, and apoptosis. PFKM overexpression mainly induced oxidative stress, ATP depletion, and increased apoptosis, whereas PFKM knockdown primarily reduced mitochondrial membrane potential, ROS, and ATP levels, indicating a relatively low-metabolic state. Both treatments were accompanied by dysregulated expression of the mitochondrial dynamics-related proteins DRP1, MFN2, and OPA1, although their patterns of change were not identical. Western blotting and immunofluorescence consistently showed that the expression levels of the myogenic differentiation markers MyoD and MYH were significantly lower in both the PFKM-KD and PFKM-OE groups than in the control group, suggesting that either excessive or insufficient PFKM expression may impair the myogenic differentiation potential of SMSCs. In conclusion, changes in PFKM expression are closely associated with glycolysis-related metabolism, energy and redox homeostasis, mitochondrial function-related indicators, and myogenic differentiation capacity in porcine SMSCs. The normal biological function of PFKM may therefore depend on its expression being maintained within an appropriate range.

Veterinary Sciences doi: 10.3390/vetsci13080741

Authors: Wentao Fei Li Yang Yuhao Zhao Zhijie Xiang Chengwei Fang Yingyu Chen Changmin Hu Aizhen Guo

Mycoplasma bovis (M. bovis) is a major pathogen in cattle, primarily responsible for bovine respiratory disease complex (BRDC). Characterized by high infectivity and morbidity, M. bovis spreads rapidly within herds and is challenging to control and eradicate, underscoring the need for early and accurate diagnosis. To address this, we developed a competitive enzyme-linked immunosorbent assay (cELISA) for detecting M. bovis-specific antibodies in bovine sera. The assay uses recombinant MbovP579 protein (1 μg/mL) as the coating antigen and an HRP-conjugated 1A2 monoclonal antibody against MbovP579 (mAb, 0.35 μg/mL) as the competitor. With a cut-off value of 40.69% (percentage inhibition, PI), the cELISA demonstrated apparent sensitivity (90%; 95% CI: 78.64–95.65%) and specificity (96.15%; 95% CI: 90.53–98.49%) using the known background positive and negative sera. In addition, it yielded 100% positivity (9/9) against sera collected from 21 days post-immunization. The assay showed no cross-reactivity with other Mycoplasma species or two common bovine bacterial pathogens, confirming its high specificity. The intra- and inter-assay coefficients of variation (CVs) were below 10%. In clinical evaluations, this cELISA generated a 55.57% positivity rate for field samples (n = 1069) and exhibited 90.46% overall agreement (967/1069; 95% CI: 88.54–92.15%) with the commercial indirect ELISA kit, demonstrating strong diagnostic consistency. Discrepant specimens were further analyzed by a metabolic inhibition test (MIT), which suggested improved specificity, although further validation is required of this cELISA.

Veterinary Sciences doi: 10.3390/vetsci13080740

Authors: Muhammet Can Cihangir Işık Cengiz Gökbulut Şehver Ege Hïsmïoğullari Gülay Turan Akın Usta Özgür Bulmuş Mustafa Hilmi Yaranoğlu

Glyphosate, the world’s most widely used herbicide, is of growing concern in both human and veterinary medicine because livestock, companion, and working animals may be chronically exposed through feed, water, and grazing on treated land. This study evaluates the protective effects of vitamin E (α-tocopherol acetate) against chronic glyphosate-induced hepatotoxicity, multi-organ histopathological damage, and disturbances of systemic redox balance in male Wistar rats. Fifty-six rats were allocated to six groups: control, low-dose glyphosate (LG, 560 mg/kg/day), high-dose glyphosate (HG, 1120 mg/kg/day), LG + vitamin E, HG + vitamin E, and vitamin E alone. Treatments were administered by oral gavage for 30 days at equalised dosing volumes. Serum oxidant/antioxidant markers—total oxidant status (TOS), total antioxidant status (TAS) and the oxidative stress index (OSI)—hepatic enzymes (AST, ALT, ALP, LDH), renal markers (urea, creatinine) and cardiac marker creatine kinase-MB (CK-MB) were measured; liver, lung, kidney, testis and heart were examined histopathologically. Glyphosate produced dose-dependent hepatocellular enzyme elevations (AST, ALT) and progressive multi-organ histopathological injury. Contrary to expectation, serum TOS and OSI did not increase in the glyphosate-only groups but were instead lower than in controls, accompanied by a compensatory rise in TAS—indicating that circulating oxidant markers may underestimate tissue-level oxidative injury at this exposure duration. Vitamin E co-administration markedly enhanced TAS, lowered TOS relative to the corresponding glyphosate group (HG + VitE vs. HG, p = 0.007), reduced OSI and substantially attenuated histopathological damage across all examined organs. Vitamin E thus mitigates glyphosate-induced tissue injury and improves systemic antioxidant capacity, supporting its potential as a protective dietary antioxidant in animals subject to chronic glyphosate exposure.

Veterinary Sciences doi: 10.3390/vetsci13080739

Authors: Rosa López-Gigosos Eduardo Martínez-Manzanares Eloísa Mariscal-López Fernando Fariñas-Guerrero Juan A. de Luque-Ibañez Jose L. Peñate-García Antonio J. Villatoro

Veterinarians are routinely exposed to occupational infectious hazards at the human–animal interface, making vaccination an important preventive measure within a One Health framework. However, evidence on vaccination coverage and immunization practices in this professional group remains limited. This study assessed self-reported vaccination coverage, attitudes toward vaccination, perceived barriers, and training needs among registered veterinarians in Málaga, Spain. A cross-sectional anonymous survey was distributed to all actively practicing veterinarians registered with the Official Veterinary Association of Málaga. The questionnaire collected sociodemographic and professional information, self-reported vaccination status, attitudes toward vaccination, and perceived training needs. A total of 164 veterinarians participated (response rate: 15.2%). Given the voluntary nature of participation, findings should be interpreted as descriptive. Vaccination coverage was low for several vaccines relevant to occupational or adult immunization, including seasonal influenza (21.3%), rabies (23.2%), and tetanus–diphtheria–pertussis boosters (47.3%). Nearly half of respondents (49.1%) were uncertain about their hepatitis B vaccination status. In contrast, COVID-19 vaccination uptake was high (96.9% received at least one dose). Although most participants considered vaccination highly important, self-reported coverage remained limited for several recommended vaccines. Qualitative responses identified barriers related to access to vaccination, information, and the absence of structured occupational vaccination programmes. Among the surveyed veterinarians in Málaga, these findings support the need for clearer occupational vaccination recommendations, improved access to immunization services, and enhanced vaccinology training. Such measures could strengthen occupational health protection and support One Health prevention strategies at the human–animal interface.

Veterinary Sciences doi: 10.3390/vetsci13080738

Authors: Muhammed Yasul Milad Afşar Ali Bilgin Yilmaz Muhammed Ahmed Selcuk Mahsa Torkamanian-Afshar Hasan Yilmaz

This study aimed to determine the prevalence of the liver trematodes Fasciola spp. and Dicrocoelium dendriticum in cattle and sheep slaughtered in Siirt Province using fecal sedimentation and postmortem liver examination to compare their detection performance, and to estimate the sensitivity of fecal sedimentation relative to postmortem examination. A total of 2211 animals, comprising 374 cattle and 1837 sheep, slaughtered during the study period, were included in the study. Fecal samples were collected rectally before slaughter, and after slaughter, the liver, gallbladder, and bile ducts were examined macroscopically. Fecal samples were analyzed using sedimentation; postmortem examination was the reference method. Based on postmortem examination, Fasciola spp. and D. dendriticum were detected in 26.2% (98/374) and 7.2% (27/374) of cattle, and in 14.8% (271/1837) and 5.9% (108/1837) of sheep, respectively. Fecal examination detected significantly fewer infections for both parasites in both hosts (McNemar test, p < 0.001 for all comparisons). The sensitivity of fecal sedimentation was 80.6% for Fasciola spp. and 29.6% for D. dendriticum in cattle, and 86.7% and 73.1%, respectively, in sheep. The prevalence of Fasciola spp. was significantly higher in cattle than in sheep (OR = 2.05, 95% CI: 1.58–2.67; p < 0.001), whereas the difference in D. dendriticum prevalence between cattle and sheep was not statistically significant (OR = 1.24, 95% CI: 0.80–1.92; p = 0.274). In cattle, age older than two years was significantly associated with Fasciola spp. (adjusted OR = 15.82, 95% CI: 8.12–30.84; p < 0.001) and D. dendriticum infection (adjusted OR = 3.29, 95% CI: 1.36–7.96; p = 0.008). In sheep, the Morkaraman breed had lower odds of Fasciola spp. infection (adjusted OR = 0.42, 95% CI: 0.32–0.56; p < 0.001) and D. dendriticum infection (adjusted OR = 0.23, 95% CI: 0.14–0.38; p < 0.001) than the Akkaraman breed, while age was significantly associated only with D. dendriticum infection (adjusted OR = 1.84, 95% CI: 1.25–2.71; p = 0.002). These findings demonstrate that liver trematode infections were common among slaughtered animals in Siirt. Reliance on fecal sedimentation may considerably underestimate prevalence, particularly for D. dendriticum in cattle. Age- and breed-related associations should be considered in surveillance and control programs. From a public health perspective, awareness of zoonotic fasciolosis and risks associated with consuming raw aquatic plants from potentially contaminated areas should be increased.

Veterinary Sciences doi: 10.3390/vetsci13080737

Authors: Michelyne Haroun Christophe Tratrat Roshmon Thomas Mathew Muhammad Munir Mohamed Shawky Ouda Nasser Aldakhilallah Mohamed Ashour Sahar Mohamed Ibrahim Athina Geronikaki

The dramatic rise in global aquaculture has brought renewed focus on filamentous fungi impacting fish health and aquafeeds. The fungal genus Aspergillus has a Janus-like role in aquaculture. On the pathogenic side, opportunistic pathogens like A. flavus, A. niger and A. fumigatus cause outbreaks of aspergillosis, with granulomatous lesions and significant mortality, especially in fish with impaired immunity and/or poor environmental conditions. Toxigenic strains of A. flavus and A. parasiticus also contaminate aquafeeds with aflatoxins, some of the most toxic natural hepatotoxins and carcinogens, which have been shown to affect growth, immunity and aflatoxin residues in edible fish fillets. On the positive side, non-toxigenic strains of A. niger, A. oryzae and A. awamori are increasingly used as fungal probiotics, solid-state fermenters to upgrade plant protein feed supplements, and as sources of industrially useful hydrolytic enzymes, secondary metabolites and antifungal compounds. Synthesizing evidence from over 90 peer-reviewed studies, this narrative review elucidates how species, strain, and rearing context jointly determine whether Aspergillus behaves as a pathogen or as a functional additive. We highlight knowledge gaps, offer an interpretative model and suggest practical strategies to limit risks and leverage the biotechnological uses of Aspergillus for sustainable aquaculture. The novelty of this review lies in integrating, within a single finfish-focused framework, the three usually separate faces of Aspergillus—pathogen, aflatoxin producer, and functional feed additive—and in translating this dual nature into practical, strain-level guidance for aquaculture.

Veterinary Sciences doi: 10.3390/vetsci13080736

Authors: Sung Su Park

A 2-year-old spayed female Bichon Frise presented with progressive neurologic deterioration despite 12 months of medical management for caudal occipital malformation syndrome (CM)-associated syringomyelia (SM) and ventriculomegaly. Repeat magnetic resonance imaging (MRI) performed prior to surgical intervention demonstrated persistent CM-associated structural abnormalities together with multifocal intra-axial lesions involving the bilateral thalami, frontal white matter, and medulla that were considered most compatible with a presumptive inflammatory meningoencephalitic process, with MUO regarded as the leading differential diagnosis. Combined surgical decompression and immunosuppressive therapy using leflunomide and mycophenolate mofetil were subsequently initiated. Progressive clinical improvement followed by long-term neurological stability was observed during the approximately 2-year follow-up period. However, because both interventions were initiated concurrently, the relative contribution of each treatment to the observed clinical outcome could not be determined. No recurrence or progression of neurological signs was observed during the approximately 2-year follow-up period. This case highlights the importance of repeat MRI evaluation in neurologic patients exhibiting progressive deterioration despite previous treatment, even when an established structural diagnosis is present. Recognition of MRI findings suggestive of concurrent inflammatory disease may substantially influence diagnostic interpretation and therapeutic decision-making in selected patients.

Veterinary Sciences doi: 10.3390/vetsci13080735

Authors: Paloma Nicolás-Barceló Jesús Fortes-Díaz Rafael Barrera Pablo Cardenal-Morales José Ignacio Cristóbal Ángela Durán Inmaculada Sevidane Francisco Javier Duque

Doxorubicin-induced cardiotoxicity is an important concern in veterinary oncology, and early detection of myocardial injury remains clinically challenging. This exploratory pilot study described changes in cardiac fatty acid-binding protein (h-FABP), cardiac troponin I (cTnI), NT-proBNP, conventional echocardiographic variables, and 24-h Holter-derived heart rate variability parameters in dogs with hemangiosarcoma treated with doxorubicin. Fourteen dogs were prospectively evaluated before treatment initiation and one week after the third doxorubicin dose, corresponding to a cumulative dose of 90 mg/m2. The study did not include a control group, a predefined cardiotoxicity endpoint, or diagnostic-performance analysis. Plasma biomarkers, echocardiographic parameters, ventricular arrhythmic events, and Holter-derived time- and frequency-domain heart rate variability variables were compared between both time points. Cardiac troponin I increased significantly after three doxorubicin doses, from a median of 0.05 ng/mL to 0.15 ng/mL (p = 0.002). In contrast, h-FABP did not change significantly, with median values of 2.43 ng/mL at baseline and 2.62 ng/mL after treatment (p = 0.27). NT-proBNP also showed no significant change (p = 0.87). Among echocardiographic variables, E-point septal separation, fractional shortening, ejection fraction, LA/Ao, and LVDd index did not change significantly. Ventricular arrhythmic events were infrequent and were described descriptively, whereas Holter-derived HRV variables did not show statistically significant differences between time points. In this exploratory cohort, cTnI increased after three doxorubicin doses, whereas h-FABP and NT-proBNP did not change significantly at the evaluated time point. Because h-FABP was measured one week after treatment, its potential short-term post-infusion release cannot be assessed. Larger studies with serial early sampling, predefined cardiotoxicity endpoints, appropriate control groups, and adjusted statistical analyses are required.

Veterinary Sciences doi: 10.3390/vetsci13080734

Authors: Li Zhang Hongxia Li Anqi Dou Changyao Fu Suyi Sun Shinuo Cao Qingkang Zhou Wei Miao Mo Zhou Wenhao Wang Jialong Xu Shanyuan Zhu

Skeletal muscle satellite cells (MuSCs) are essential for muscle growth and development, but their regulatory mechanisms remain unclear. This study aimed to characterize the expression pattern of pleomorphic adenoma gene 1 (PLAG1), determine its effects on MuSC proliferation and differentiation, and explore its potential regulatory mechanisms through integrated transcriptomic and ceRNA analyses in Sujiang pig MuSCs. Results showed that PLAG1 was widely expressed in multiple porcine tissues and was significantly upregulated during MuSC differentiation. PLAG1 overexpression promoted MuSC proliferation and differentiation, whereas PLAG1 knockdown produced the opposite effects. Whole-transcriptome sequencing following PLAG1 knockdown identified 432 differentially expressed mRNAs, 70 miRNAs, 163 lncRNAs, and 150 circRNAs. Enrichment analysis revealed that differentially expressed mRNAs were mainly associated with cell cycle regulation, DNA replication, and the p53 signaling pathway. Western blot analysis further showed that PLAG1 knockdown reduced CDK1 and PCNA protein levels. ceRNA network analysis revealed potential regulatory associations among differentially expressed lncRNAs, circRNAs, miRNAs, and mRNAs, and the expression changes in selected differentially expressed RNAs were further validated by qRT-PCR. Collectively, these findings indicate that PLAG1 promotes MuSC proliferation and differentiation and may be associated with cell cycle-related gene expression changes and potential non-coding RNA regulatory networks in porcine MuSCs.