Veterinary Sciences doi: 10.3390/vetsci13070682

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

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

Veterinary Sciences doi: 10.3390/vetsci13070681

Authors: Jihee Han Sijin Cha Jeonghyun Seo Kunho Song

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

Veterinary Sciences doi: 10.3390/vetsci13070680

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

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

Veterinary Sciences doi: 10.3390/vetsci13070679

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

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

Veterinary Sciences doi: 10.3390/vetsci13070678

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

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

Veterinary Sciences doi: 10.3390/vetsci13070677

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

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

Veterinary Sciences doi: 10.3390/vetsci13070676

Authors: Renato Leon Amanda Blubaugh Haley Starr Frane Banovic

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

Veterinary Sciences doi: 10.3390/vetsci13070675

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

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

Veterinary Sciences doi: 10.3390/vetsci13070674

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

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

Veterinary Sciences doi: 10.3390/vetsci13070673

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

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

Veterinary Sciences doi: 10.3390/vetsci13070672

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

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

Veterinary Sciences doi: 10.3390/vetsci13070671

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

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

Veterinary Sciences doi: 10.3390/vetsci13070670

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

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

Veterinary Sciences doi: 10.3390/vetsci13070669

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

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

Veterinary Sciences doi: 10.3390/vetsci13070668

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

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

Veterinary Sciences doi: 10.3390/vetsci13070667

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

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

Veterinary Sciences doi: 10.3390/vetsci13070666

Authors: Annika Gersemann Gerald Reiner Sonja von Berg

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

Veterinary Sciences doi: 10.3390/vetsci13070665

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

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

Veterinary Sciences doi: 10.3390/vetsci13070664

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

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

Veterinary Sciences doi: 10.3390/vetsci13070663

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

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

Veterinary Sciences doi: 10.3390/vetsci13070662

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

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

Veterinary Sciences doi: 10.3390/vetsci13070661

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

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

Veterinary Sciences doi: 10.3390/vetsci13070659

Authors: Aaron C. Ericsson

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

Veterinary Sciences doi: 10.3390/vetsci13070660

Authors: Rhoda C. Apitzsch Sarah Handel Konstanze Krueger

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

Veterinary Sciences doi: 10.3390/vetsci13070658

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

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

Veterinary Sciences doi: 10.3390/vetsci13070657

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

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

Veterinary Sciences doi: 10.3390/vetsci13070656

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

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

Veterinary Sciences doi: 10.3390/vetsci13070655

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

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

Veterinary Sciences doi: 10.3390/vetsci13070654

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

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

Veterinary Sciences doi: 10.3390/vetsci13070653

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

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

Veterinary Sciences doi: 10.3390/vetsci13070652

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

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

Veterinary Sciences doi: 10.3390/vetsci13070651

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

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

Veterinary Sciences doi: 10.3390/vetsci13070650

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

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

Veterinary Sciences doi: 10.3390/vetsci13070649

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

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

Veterinary Sciences doi: 10.3390/vetsci13070648

Authors: Kyuman Cho

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

Veterinary Sciences doi: 10.3390/vetsci13070647

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

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

Veterinary Sciences doi: 10.3390/vetsci13070646

Authors: Ericka L. Carroll Nouran Nawar Melissa Bourgeois

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

Veterinary Sciences doi: 10.3390/vetsci13070645

Authors: Andrés Montesinos Barceló Alicia Tortosa García María Ardiaca García

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

Veterinary Sciences doi: 10.3390/vetsci13070644

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

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

Veterinary Sciences doi: 10.3390/vetsci13070643

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

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

Veterinary Sciences doi: 10.3390/vetsci13070642

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

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

Veterinary Sciences doi: 10.3390/vetsci13070641

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

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

Veterinary Sciences doi: 10.3390/vetsci13070640

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

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

Veterinary Sciences doi: 10.3390/vetsci13070639

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

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

Veterinary Sciences doi: 10.3390/vetsci13070638

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

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

Veterinary Sciences doi: 10.3390/vetsci13070637

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

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

Veterinary Sciences doi: 10.3390/vetsci13070636

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

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

Veterinary Sciences doi: 10.3390/vetsci13070635

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

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

Veterinary Sciences doi: 10.3390/vetsci13070634

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

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

Veterinary Sciences doi: 10.3390/vetsci13070633

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

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

Veterinary Sciences doi: 10.3390/vetsci13070632

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

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

Veterinary Sciences doi: 10.3390/vetsci13070631

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

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

Veterinary Sciences doi: 10.3390/vetsci13070630

Authors: Zarroug Hassan Ibrahim

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

Veterinary Sciences doi: 10.3390/vetsci13070629

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

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

Veterinary Sciences doi: 10.3390/vetsci13070627

Authors: Indra Sara Klumb Axel Wehrend Abbas Farshad

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

Veterinary Sciences doi: 10.3390/vetsci13070628

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

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

Veterinary Sciences doi: 10.3390/vetsci13070626

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

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

Veterinary Sciences doi: 10.3390/vetsci13070624

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

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

Veterinary Sciences doi: 10.3390/vetsci13070625

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

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

Veterinary Sciences doi: 10.3390/vetsci13070623

Authors: Mario Baratta Paolo Accornero Silvia Miretti Eugenio Martignani

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

Veterinary Sciences doi: 10.3390/vetsci13070622

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

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

Veterinary Sciences doi: 10.3390/vetsci13070621

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

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

Veterinary Sciences doi: 10.3390/vetsci13070620

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

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

Veterinary Sciences doi: 10.3390/vetsci13070619

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

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

Veterinary Sciences doi: 10.3390/vetsci13070618

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

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

Veterinary Sciences doi: 10.3390/vetsci13070617

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

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

Veterinary Sciences doi: 10.3390/vetsci13070616

Authors: Wei Han Zhao Si Yeong Choi In Ho Kim

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

Veterinary Sciences doi: 10.3390/vetsci13070615

Authors: Begoña Acosta-Hernández Nicolás Cabrera Guerle Pablo Lorenzo García Olga Armas Carballo María del Mar Ojeda-Vargas Victor Garcia-Bustos Fernando Real Valcárcel Soraya Déniz Suárez Esther Licia Díaz Rodríguez Inmaculada Rosario Medina

Wild and synanthropic birds harbour a diverse range of yeasts, including species of recognised clinical relevance. Given their close interaction with human activities, these birds represent a valuable source for investigating environmental yeasts and assessing the performance of identification methods. We investigated yeasts recovered from cloacal and crop samples of birds from Gran Canaria and compared routine identification methods with molecular sequencing. Twenty-four isolates were examined by biochemical profiling (API ID32C) and MALDI-TOF MS. Molecular identification based on ITS sequencing was carried out only for the sixteen isolates for which the two routine methods yielded discordant results, allowing resolution of their taxonomic identification. Phenotypic and proteomic methods showed limited agreement at the species level (8/24; 33.3%), with 16 discordant identifications resolved by sequencing. Pigeon isolates were dominated by members of the Kazachstania telluris complex, chiefly K. bovina (11/24; 45.8%), while partridges yielded taxa of clinical importance, including Candida parapsilosis and Pichia kudriavzevii (formerly Candida krusei). Rhodotorula mucilaginosa, Debaryomyces spp., and Saccharomyces cerevisiae were also detected. Comparative tests confirmed significant host-associated differences in species distribution (p < 0.05), and Cohen’s kappa indicated substantial agreement between API and MALDI-TOF at the genus level when benchmarked against ITS (κ = 0.71), although concordance was lower at the species level. In conclusion, these findings strengthen the case for integrating sequencing into diagnostic workflows, highlight the potential One Health relevance of yeast carriage by wild birds, and underscore the need for targeted surveillance at urban and game-handling interfaces where human exposure is likely.

Veterinary Sciences doi: 10.3390/vetsci13070614

Authors: Jia-Qi Zhang Jia-Xin Li Hui-Lin Qu Yu-Jie Miao Xi-Meng Chen Lan-Lan Zheng Yi-Lei Li Hong-Ying Chen Shi-Jie Ma

Porcine circovirus type 5 (PCV5) is a recently reported porcine-associated CRESS DNA virus, but information regarding its occurrence, genomic characteristics, and evolutionary relationship remains limited. In this study, a total of 100 diagnostic samples collected from clinically diseased pigs from 27 commercial swine farms in 16 cities across seven provinces of China during 2025 were screened for PCV5 using quantitative PCR. PCV5 was detected in 22% (22/100) of the tested samples, with positive samples identified in Henan and Fujian provinces among the sampled regions. PCV5-positive samples were mainly fecal samples and were obtained from pigs showing diarrhea, respiratory signs, wasting, or systemic disease. Co-detection analysis showed that most PCV5-positive samples were also positive for other swine viral pathogens, particularly PCV3, PCV2, and PEDV, indicating that the clinical significance of PCV5 should be interpreted cautiously. Complete genome amplification and sequencing yielded two identical PCV5 genomes from Henan and Fujian provinces. A representative strain, designated PCV5-Henan2025-ZJQ01, was further characterized and deposited in GenBank under accession number PZ496079. The complete genome was 2903 nt in length and contained a positive-sense ORF encoding Cap and a negative-sense ORF encoding Rep, showing a distinct genomic organization compared with classical porcine circoviruses. Phylogenetic analysis based on Rep and Cap amino acid sequences showed that PCV5-Henan2025-ZJQ01 was closely related to previously reported PCV5-related sequences but distinct from classical PCV1–PCV4. These findings provide additional molecular and genomic evidence for PCV5 in Chinese swine diagnostic samples and support the need for continued surveillance and further studies on its epidemiological and pathogenic significance.

Veterinary Sciences doi: 10.3390/vetsci13070613

Authors: Qian Ma Jiaqi Dong Rong Yang Yongli Hua Fanlin Wu Yanming Wei Peng Ji

This study investigated the protective effects of DBD against Staphylococcus aureus (S. aureus)-induced mastitis in mice and explored whether these effects were associated with gut microbiota alterations, blood–milk barrier integrity, and inflammatory signaling. A lactating mouse model of mastitis was established, and the effects of DBD were evaluated using HPLC, histopathological analysis, ELISA, qRT-PCR, Western blotting, immunofluorescence, and 16S rRNA sequencing. The results showed that DBD significantly reduced bacterial loads in mammary tissues, decreased the expression of pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6, and alleviated inflammatory cell infiltration and tissue damage. Moreover, DBD upregulated the expression of tight junction proteins and improved the integrity of the blood–milk barrier. DBD treatment was also associated with alterations in gut microbiota composition, as reflected by changes in the relative abundance of several bacterial taxa. In addition, DBD inhibited the activation of the NF-κB/NLRP3 and MAPK inflammatory signaling pathways. Collectively, these findings indicate that DBD alleviates S. aureus-induced mastitis accompanied by alterations in gut microbiota composition, suppressing inflammatory responses, and repairing the blood–milk barrier, suggesting its potential as a therapeutic agent for mastitis.

Veterinary Sciences doi: 10.3390/vetsci13070612

Authors: Nadia Piscopo Esterina De Carlo Anna Cerrone Domenico Vecchio Michele Napoletano Agata Campione Chiara Denise Ambra Michael J. D’Occhio Marco Esposito Alessio Cotticelli Tanja Peric Giuseppe Campanile

Brucellosis negatively impacts animal and human health and causes major economic losses. The present study examined the relationship of husbandry system (insufficient, acceptable, optimal) and cortisol status on the response of buffalo calves to vaccination with the brucellosis vaccine RB51. Primary vaccination (1st) was at 6 months and secondary vaccination (2nd) at 12 months. Cortisol was ascertained in samples of hair and the antibody response to vaccination was measured in blood. Excretion of the bacterium Brucella abortus was determined in urine and feces. Buffalo calves in the insufficient system had greater (p < 0.05) cortisol (4.7 ± 0.5 pg/mg) at 1st vaccination than calves in the optimal system (2.8 ± 0.3 pg/mg). Cortisol was lower at the 2nd vaccination for all three systems which did not differ. Anti-RB51 antibodies were present in 28/30 buffalo after 2nd vaccination. The conditional R2 of 0.689 indicated that both husbandry system and cortisol status strongly influenced the response to vaccination with RB51. No shedding of Brucella spp. was observed. The study has shown that husbandry system and cortisol status influence the response of buffalo calves to vaccination with RB51.

Veterinary Sciences doi: 10.3390/vetsci13070611

Authors: Ibtissem Boulbina Mohammed El-Amine Bekara Hacina AinBaziz Asma Kassoul Cesare Castellini

This study aimed to characterize the reproductive performance of the local Algerian population (LAP) compared with the New Zealand White (NZW) rabbits, by evaluating sexual behavior, semen characteristics, and their modulation by environmental factors, namely photoperiod and temperature-humidity index (THI). Mature bucks (n = 14/breed) were monitored from January to April, with two successive ejaculates collected weekly. Sexual behavior, macroscopic and microscopic semen parameters, and testosterone concentrations were assessed. The effects of breed, ejaculate order, environmental factors, and their interactions were analyzed using Generalized Linear Mixed models. LAP and NZW bucks exhibited similar sexual behavior and blood testosterone levels (p > 0.05). Collection failures and ejaculate rejection causes were mainly clustered within specific individuals rather than being breed-dependent. However, LAP bucks showed higher sperm concentration (p = 0.01), viability (p = 0.02), and membrane integrity (p = 0.04) than NZW bucks, whereas most motility and quantitative semen traits remained comparable between breeds. Increasing photoperiod significantly improved reproductive performance (p < 0.05). Conversely, within the investigated range, THI mainly affected semen collection efficiency through increased urine contamination (p < 0.001), with limited effects on intrinsic sperm quality. Significant breed × environment interactions for sperm concentration (p = 0.03) suggested differential responsiveness between breeds, with LAP bucks showing a stronger positive response to increasing photoperiod and less pronounced variation under THI fluctuations. Overall, LAP bucks exhibited a more favorable seminal profile under the conditions of the present study, supporting the valorization of this local genetic resource for artificial insemination programs under Algerian conditions. Further studies are required to confirm these patterns under summer heat-stress conditions and evaluate their impact on fertility outcomes.

Veterinary Sciences doi: 10.3390/vetsci13070610

Authors: Yuan Chai Luxin Lou

Traumatic fractures are common in small animal emergency care, yet subtle fracture lines may be difficult to identify accurately using routine three-dimensional reconstruction workflows, particularly when access to specialized software is limited. This study describes the use of the open-source platform Three-Dimensional Slicer for computed tomography-based reconstruction and three-dimensional printing in a small dog with cranial trauma, with emphasis on documenting a practical and reproducible workflow through voxel resampling. Imaging data were imported into the software, bone structures were segmented using a rapid workflow, voxel spacing was resampled for smoother surface visualization by volume resampling, and the reconstructed model was processed for physical printing. Digital models of different resolutions were generated within minutes, and a life-size skull model was successfully fabricated using fused deposition modeling in less than three hours at a material cost of under one United States dollar. The enhanced model provided an intuitive representation of fracture morphology and spatial relationships compared with routine reconstruction alone. These findings demonstrate that open-source software combined with low-cost printing can provide a rapid, affordable, and user-friendly approach for practical skeletal reconstruction in small animals, with practical value for fracture assessment, preoperative planning, and broader use in resource-limited veterinary settings.

Veterinary Sciences doi: 10.3390/vetsci13070609

Authors: Shuaishuai Wu Mohamed Tharwat Ibrahim F. Halawani Fuad M. Alzahrani Khalid J. Alzahrani Muhammad Zahoor Khan

Mastitis remains the most economically damaging disease of dairy production, and recent molecular work has converged on the NLRP3 inflammasome as a key integrative node of its pathogenesis. This narrative review integrates evidence published largely between 2015 and 2026 to show how diverse triggers—Staphylococcus aureus and Escherichia coli, lipopolysaccharide (LPS) and lipoteichoic acid (LTA), non-esterified fatty acids (NEFA), heat stress, environmental xenobiotics including nanoplastics, and microbiota-derived signals—may funnel into a common NLRP3–ASC–caspase-1–GSDMD axis that drives pyroptosis, blood–milk barrier disruption, and clinical disease. The review examines the potential obligatory role of reactive oxygen species (ROS), mitochondrial dysfunction, and selenoprotein-mediated redox control in licensing inflammasome assembly. It further evaluates the emerging gut–mammary and rumen–mammary axes that operate upstream of local epithelial activation. We survey a structurally diverse therapeutic landscape encompassing dietary selenium, probiotics, microbial metabolites, plant-derived nanovesicles, polyphenols, ginsenosides, and small-molecule NLRP3 antagonists, identifying recurring mechanistic motifs that suggest combinatorial regimens may yield additive benefit. Importantly, much of the evidence derives from in vitro and murine models, and we highlight the translational gaps that must be bridged before clinical application in dairy cattle. Finally, we map outstanding research gaps and propose priorities for translational work aimed at sustainable, antibiotic-sparing management of bovine mastitis.

Veterinary Sciences doi: 10.3390/vetsci13070608

Authors: Leyao Xu Jianfa Wang Rui Wu

In dairy cows, teat keratinization occurs where keratinized tissue rings form around teat tips (opening). Manifesting as skin damage in mechanically milked dairy cows, keratinization presents as dry, rough, pale, or milky-white keratinous protrusions around teat orifices and progresses via the combined effects of mechanical stress, management practices, host genetics, environmental influences, and nutritional metabolism. The teat hole is the first physiological barrier protecting the mammary glands from external pathogen invasion. Most mastitis cases are caused by pathogens invading the mammary tissues from the teat end, thereby significantly impacting mammary gland health. Thus, there are no safe, effective, economical, and standardized treatment protocols for terminal teat hyperkeratosis in dairy cows. To address this, we treated keratinization lesions using pharmacological interventions and evaluated their efficacy in 91 cows at a large commercial dairy farm in Heilongjiang Province. Urea ointment, salicylic acid ointment, and 5% azelaic acid acne cream exerted therapeutic or alleviating effects toward teat keratosis, whereas retinoic acid ointment demonstrated poor therapeutic efficacy. In a three-daily application regimen, the optimal treating dose was 0.3 g. These findings provide a scientific basis for the clinical management of hyperkeratosis in large-scale dairy farms and the development of related veterinary therapeutics.

Veterinary Sciences doi: 10.3390/vetsci13070607

Authors: Isabela Cristina Colaço Bez Ana Julia Carrasco Buzatto Claudia Sevillano Marcos Lopes Saulo Henrique Weber Leandro Batista Costa

Seasonal infertility remains a major challenge in pig production and is influenced by environmental conditions and sow parity. This study aimed to identify in-barn heat load (HL), light variability (LV), and parity as farm-specific predictors of farrowing success (FS) and litter size (LS) in purebred Large White sows raised under subtropical and tropical conditions in Brazil. Reproductive records from 2021 were obtained from two commercial farms, and in-barn temperature and illuminance were recorded using data loggers installed in gestation units. Services between March and August 2021 were analyzed using logistic regression for FS and linear regression for LS, resulting in 732 service records from Farm 1 and 233 from Farm 2. In the combined analysis, parity was associated with higher odds of FS (OR = 1.20; p = 0.003) and greater LS (β = 0.47; p < 0.001). HL was not associated with FS but showed a small positive association with LS (β = 0.28; p = 0.031). In Farm 2, LV was negatively associated with FS (OR = 0.72; p = 0.028) and LS (β = −0.59; p = 0.020). Overall, parity structure strongly shaped reproductive performance, and in-barn environmental effects were context-dependent.

Veterinary Sciences doi: 10.3390/vetsci13070606

Authors: Yongwon Park Ajin Lee Jeonghoon Jang Hwi-Yool Kim

Apocrine sweat gland adenocarcinoma is an uncommon feline adnexal neoplasm, and lower-lip involvement is rarely documented. A 14-year-old spayed female Siamese cat was evaluated for a firm 1.3 cm right lower-lip mass at a documented hemorrhagic lesion site. Approximately 1 year earlier, the lesion was fluctuant and superficially erosive to ulcerative; fine-needle aspiration yielded bloody fluid with cytologically nondiagnostic material. The lesion reportedly subsided after empirical treatment, but reassessment, tissue diagnosis, and objective confirmation of resolution were not performed. Cytology of the later same-site mass supported malignant epithelial neoplasia. Contrast-enhanced computed tomography (CT) identified a 12.5 × 13.2 × 16.6 mm rim-enhancing mass without mandibular invasion or distant metastasis; the ipsilateral mandibular lymph node was mildly thickened. Histopathology supported a diagnosis of lower-lip apocrine sweat gland adenocarcinoma with lymphovascular tumor emboli, lymphatic invasion, and nodal metastasis. The diagnosis was supported by superficial dermal/subcutaneous location, adjacent adnexal structures, skeletal-muscle separation from nearby minor salivary glands, epithelial immunoreactivity, periodic acid-chiff (PAS) and Alcian blue findings without distinct intratumoral mucin/mucopolysaccharide-rich material. The cat died without necropsy on postoperative day 87. This case supports tissue-based diagnosis and regional lymph node assessment for suspicious feline lower-lip glandular epithelial lesions.

Veterinary Sciences doi: 10.3390/vetsci13070605

Authors: José Luis Granados-Soler Michelle Majella Story Rachel Allavena

Ultraviolet (UV) light exposure is a recognised risk factor for dermal haemangiosarcoma (HSA) in dogs and dermal squamous cell carcinoma (SCC) in dogs and cats. These tumours cause substantial local disease and often require repeated surgery due to recurrence or de novo lesions, creating a notable welfare and financial burden. Research on preventing harmful UV exposure in pets is still in its infancy, particularly in relation to the safety of UV filters used in sunscreens. This review summarises the current evidence on UV-induced carcinogenesis, strategies to reduce UV exposure, and the safety of sunscreen ingredients in dogs and cats. UV light is strongly implicated in a range of dermatoses, from actinic keratosis to dermal HSA and SCC in dogs and cats, and the risk is likely higher in Australian pets. Indoor confinement during peak UV periods, shade, and sun-protective clothing can reduce exposure, with sunscreen an additional strategy. Sunscreen is relevant because UV-associated cancers typically develop in sparsely haired or hairless regions such as the nose and ventrum, making these areas suitable for targeted sunscreen application. Sunscreens containing non-nanoparticle zinc oxide appear safe for dogs and cats when ingestion is prevented or minimised, whereas the safety of organic UV filters remains unclear due to limited safety data in both humans and animals. Non-nanoparticle titanium dioxide is a possible alternative to zinc oxide and organic filters, but there is currently little information on its safety when ingested by dogs and cats. Overall, the available evidence supports sunscreen as a necessary component of UV-reduction strategies in pets, but substantial research is needed to determine the safety profiles of different UV filters and to establish evidence-based guidelines for their safe use.

Veterinary Sciences doi: 10.3390/vetsci13060604

Authors: Cleverson de Souza

Chronic kidney disease (CKD) is common in older cats, but cortical transcript-level relationships among hypoxia response, iron handling, and lipid oxygenation are poorly defined. This analysis-plan-guided, hypothesis-generating secondary analysis used public feline renal RNA-seq data (GSE303653). The internal plan fixed the gene panel, composite construction, primary inferential test, and quality-control thresholds before the review of the present expression results, but was not publicly registered. After technical quality control, 21 renal cortex samples from control, CKD 1/2, and CKD 3/4 cats were analyzed. A 23-gene panel and whole-transcriptome differential expression were evaluated using likelihood ratio testing as the primary panel-level screen, with pairwise DESeq2 contrasts, Spearman summaries, enrichment, medulla, composite, and marker-set analyses as secondary or exploratory context. VEGFA, FTL, and NCOA4 decreased with ordinal disease group, whereas ALOX5 and HIF1A increased; eight panel genes were stage-associated by likelihood ratio testing. The equal-weight composite was nonmonotonic. Advanced CKD enrichment was dominated by immune and inflammatory terms, while GPX4 and ferroptosis-pathway enrichment were not stage-significant. The findings support heterogeneous transcript-level remodeling, including ALOX5-associated inflammatory/lipid-oxygenation signal and HIF1A–VEGFA divergence, rather than evidence of ferroptotic cell death, pathway activation, or cell-specific mechanism.

Veterinary Sciences doi: 10.3390/vetsci13060603

Authors: Waresi Tuersong Lianxi Xin Abudusaimaiti Tuoheti Ailixire Maimaiti Dilare Xuekelaiti Reyilanmu Tuerhong Wei Zhang Bayinchahan Gailike Qingyong Guo Saifuding Abula

Background: Benzimidazole (BZ) resistance in the gastrointestinal nematode Haemonchus contortus is a major constraint to sheep production worldwide. However, data on the prevalence and molecular mechanisms of resistance in Yili Prefecture, Xinjiang—a key livestock region in Northwestern China—remain limited. This study aimed to determine the frequency of BZ resistance-associated single nucleotide polymorphisms (SNPs) in H. contortus populations from Zhaosu and Tekesi counties. Methods: Adult male worms (n = 150) were collected from naturally infected sheep at local abattoirs. Species identity was confirmed morphologically by sequencing the internal transcribed spacer 2 (ITS-2) region. A 385 bp fragment of the isotype-1 β-tubulin gene was amplified and sequenced to detect SNPs at codons 167 (F167Y), 198 (E198A), and 200 (F200Y). Results: The F167Y mutation was absent in all individuals. In contrast, the E198A mutation occurred at exceptionally high frequencies, with resistant allele frequencies (RAF) of 64.7% in Zhaosu and 52.7% in Tekesi. The F200Y mutation showed clear geographical variation: it remained low in Zhaosu (RAF = 9.3%) but was substantially higher in Tekesi (RAF = 33.3%). Haplotype analysis revealed that resistance in Zhaosu was driven primarily by the E198A mutation, whereas the Tekesi population exhibited complex patterns of co-selection of both E198A and F200Y, with a high proportion of double-heterozygous individuals (29.3%). Conclusions: This study provides comprehensive molecular evidence of severe BZ resistance in H. contortus populations from Zhaosu and Tekesi counties, Yili Prefecture. The marked predominance of the E198A mutation, together with the emergence of multi-locus resistance in Tekesi, indicates a rapid escalation of resistance beyond historical levels. These findings suggest that benzimidazoles are likely ineffective in this region and highlight the urgent need to revise local parasite control strategies.

Veterinary Sciences doi: 10.3390/vetsci13060602

Authors: Dequan Guo Xin Fan Zijie Lan Chengli Zheng Dapeng Zhang Zhenyu Wang Minyao Tan

In artificial breeding of forest musk deer (Moschus berezovskii), common diseases such as abscess, enteritis, pneumonia, and parasitic infections exhibit persistently high morbidity rates. The early symptoms of certain diseases are often insidious and difficult to discern. Conventional manual inspection routines not only fail to achieve accurate diagnosis but also frequently disturb the animals, induce stress responses, and consequently delay optimal treatment windows. To address this practical challenge, this study employs an improved BRW-GPLinker joint entity-relationship extraction approach to perform integrated extraction and structural organization of disease entities, symptom manifestations, etiological associations, and preventive and therapeutic measures from farming literature and clinical records, thereby constructing a disease knowledge graph for forest musk deer. Through the introduction of a Boundary-Aware Module for refined entity boundary detection, a Relative Distance Bias Module to mitigate pairing errors in dense contexts, and a Weighted Sparse Multi-label Cross-Entropy loss function to enhance recall for infrequent relations, the proposed model achieves an F1 score of 0.887 on a self-constructed dataset and demonstrates favorable generalization capability on medical-domain datasets. By transforming fragmented clinical logs and manuals into structured medical associations, this knowledge graph facilitates rapid retrieval of forest musk deer disease information, thereby enhancing veterinary decision-making efficiency and assisting forest musk deer health management.

Veterinary Sciences doi: 10.3390/vetsci13060601

Authors: Akihiro Ohnishi Yuko Mizutani Saki Kageyama Shinya Mizutani Taketoshi Asanuma

We described tumor response and survival in dogs with modified Adams stage 4 intranasal tumors treated with once-weekly hypofractionated radiation therapy (RT) combined with the radiosensitizer α-sulfoquinovosyl-acylpropanediol (SQAP), and compared linear and volumetric response assessments. Twenty dogs treated with intensity-modulated RT (8 Gy per fraction, once weekly) and concurrent SQAP were included in this retrospective case series. Tumor response was assessed using RECIST-like linear measurements and volumetric analysis on contrast-enhanced computed tomography. Overall survival (OS) was estimated using Kaplan–Meier analysis. Of the 20 dogs, 4 were classified as stage 4a and 16 as stage 4b. The best RECIST-like responses were complete response (CR) in 5 dogs, partial response (PR) in 12, and stable disease (SD) in 4. Volumetric responses were CR in 5 dogs, PR in 11, and SD in 5. No cases demonstrated progressive disease as the best response. The median OS for all dogs was 342 days (95% confidence interval [CI], 206–419 days). Censoring one non-tumor-related death yielded a median OS of 356 days (95% CI, 231–419 days). Exploratory analysis revealed median OS of 393 and 297 days for stage 4a and 4b dogs, respectively. Volumetric assessment appeared more sensitive for detecting tumor regrowth in selected cases. Dermatologic adverse events were limited to alopecia within the radiation field, and no complete vision loss was observed. Seizure activity was documented in eight dogs. In conclusion, once-weekly hypofractionated intensity-modulated RT combined with concurrent SQAP was associated with clinically meaningful survival outcomes in dogs with advanced intranasal tumors. However, because no radiotherapy-alone control group was available, the independent contribution of SQAP to these outcomes could not be determined.

Veterinary Sciences doi: 10.3390/vetsci13060600

Authors: Andrea García-Mendoza Milagros González-Hernández Delia X. Vega-Manriquez Erika Félix-Santiago María del Refugio Pérez-Barba César A. Rosales-Nieto

This study investigated the associations among dam lactation number, passive immunity transfer, internal parasite burden, and weaning performance in female calves. Early consumption of high-quality colostrum is critical for effective passive immunity transfer, calf health, and long-term productivity. Colostrum composition may vary with management practices and dam lactation number, potentially influencing immunity and disease susceptibility. Forty-five cows and their female offspring (n = 45) were allocated to three groups by lactation number: first lactation (LAC1), second lactation (LAC2), and third lactation (LAC3) (15 cows and 15 calves per group). Calf birth weight was recorded. Colostrum immunoglobulin G (IgG) concentration was measured using a colostrometer and refractometer, and chemical composition (fat, protein, and non-fat solids) was analyzed. Fecal samples collected at 30, 60, and 90 days of age were examined for Eimeria spp. Colostrum IgG concentration, °Brix percentage, specific gravity, and non-fat solids did not differ among lactation groups (p > 0.05). LAC3 cows had higher colostrum protein content, while LAC1 cows had greater fat concentration (p < 0.05). Calves from LAC1 dams were lighter at birth than those from LAC2 and LAC3 dams (p < 0.001). The prevalence of Eimeria spp. was not influenced by lactation number, birth weight, or colostrum quality (p > 0.05), but treatment costs were higher in calves from LAC3 dams (p < 0.01). In conclusion, lactation number affected colostrum composition and calf growth but did not alter IgG concentration, underscoring the importance of effective colostrum management to improve calf performance and dairy system sustainability.

Veterinary Sciences doi: 10.3390/vetsci13060599

Authors: Yangyan Yin Changting Li Yongcui Feng Huili Bai Zhe Pei Zhongwei Chen Yanwen Zhang Caifeng Li Chunxia Ma Dongyan Deng Leping Wang Ling Teng Hailan Chen Hao Peng

This study investigated the effects of composite postbiotic preparation on the growth performance, immune function, and microbiota composition of Nubian black goats. Thirty healthy Nubian black goats with similar body weights were randomly assigned to two groups (n = 15 per group): a control group fed a basal diet and a treatment group fed the basal diet supplemented with 0.5% composite postbiotic preparation (equal-ratio co-fermentation of Bacillus subtilis GX15 and Lentilactobacillus buchneri GX0328-6). The results indicated that while compound postbiotic supplementation did not significantly alter the average daily gain (ADG) and the serum biochemical indices (p > 0.05), it significantly increased the concentrations of immunoglobulins (IgG and IgA) compared to the control group (p < 0.05). Despite comparable α- and β-diversity, CPP supplementation selectively enriched Bacteroides, UCG-005, and Ruminococcaceae while reducing Turicibacter (LEfSe LDA > 2.0; STAMP p < 0.05), suggesting targeted modulation of gut microbiota. In conclusion, dietary supplementation with 0.5% composite postbiotic preparation improves immune function and modulates intestinal microbiota composition without significantly affecting growth performance in black goats while improving intestinal microbial composition and promoting overall gut health.

Veterinary Sciences doi: 10.3390/vetsci13060598

Authors: Tahani M. I. Al-Hazani Amirah S. Alahmari Manal A. Babaker Ahmed M. Elbaz Hagar E. Mohammed Hany A. Thabet Eman Kamel M. Khalfallah Ahmed Ateya Rowa K. Zarah Khairiah Mubarak Alwutayd Assem Abdou

This study evaluated the effects of adding zinc oxide nanoparticles (ZnNP), L-arginine (L-Arg), or a combination of both to the diets of growing rabbits to mitigate the physiological and productive consequences of heat stress. Two hundred and eighty 35-day-old New Zealand White rabbits were randomly assigned to four experimental treatments, with 70 rabbits per treatment and seven replicates (10 rabbits/replicate). The control group (Ctr) received the base diet without additives, while the diets of the other groups were fortified with arginine (L-Arg; 3 g/kg), zinc oxide nanoparticles (ZnNP; 40 mg/kg), or a combination of both (Arg-Zn). The results showed that the combined Arg-Zn significantly improved weight gain rate, feed conversion rate, carcass weight, and nutrient digestibility compared to the control group (p < 0.05). At the physiological level, we observed increased serum levels of total antioxidant capacity (T-AOC), glutathione peroxidase (GPx), superoxide dismutase (SOD), immunoglobulin G (IgG), immunoglobulin A (IgA), and triiodothyronine (T3), along with decreased levels of malondialdehyde (MDA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST, p < 0.05) in Arg-Zn-fed rabbits. However, adding the Arg-Zn mixture contributed to a reduction in pathogenic bacteria counts and increased the volatile fatty acid (VFA) levels. At the molecular level, the gene expression of the inflammatory cytokines IL-6 and tumor necrosis factor alpha (TNF-α) decreased; however, the gene expression of claudins-1 (CLDN-1), cationic amino acid transporter-1 (CAT-1), mucin-2 (MUC-2), sodium-glucose co-transporter-1 (SGLT-1), and interferon gamma (IFNγ) increased (p < 0.05) in Arg-Zn-fed rabbits. These results suggest that dietary supplementation with ZnNP and L-Arg may serve as an effective nutritional strategy for improving growth performance, antioxidant status, immune function, and intestinal integrity in rabbits exposed to high ambient temperatures.

Veterinary Sciences doi: 10.3390/vetsci13060597

Authors: Soontaree Petchdee Xufeng Ying Suchada Huttayananont Kotchapol Jaturanratsamee Chattida Panprom Wannisa Meepoo Ratikorn Bootcha

Transcatheter edge-to-edge repair (TEER) is a recent minimally invasive method of managing mitral regurgitation (MR) in dogs with myxomatous mitral valve disease (MMVD). As the goal of intervention is to minimize MR severity, this study aimed to determine the association between reduced MR and changes in myocardial work indices after TEER in dogs. Ten client-owned dogs with moderate-to-severe MR were enrolled in the study, and all underwent TEER with multimodal imaging guidance. Myocardial work was analyzed before and after the procedure, and the MR severity, transmitral pressure gradients, left atrial and ventricular measurements, and index of myocardial work (GWI: the total myocardial work during systole; GCW: work contributing to LV ejection; GWW: ineffective work that contributes to no forward displacement; and GWE: ratio of constructive work to total work) were calculated. TEER significantly reduced MR severity in the majority of dogs, and this MR decrease was associated with a greater efficiency of myocardial work, more constructive work, and less wasted energy. No significant negative associations of moderate post-procedure gradients with short-term clinical outcomes emerged. TEER-mediated reduction in MR improves myocardial function in dogs. However, long-term studies are also needed to examine the effects of residual MR and transmitral gradients on cardiac function and clinical outcome.

Veterinary Sciences doi: 10.3390/vetsci13060596

Authors: Asumi Muramatsu Tomokazu Nagashima Kazuhiko Ochiai Amo Ohnuma Honoka Kawamura Yukino Machida Daigo Azakami Makoto Bonkobara Toshiyuki Ishiwata Masaki Michishita

Primary pulmonary adenocarcinoma in dogs is rare, and effective systemic therapies remain limited. To evaluate the molecular basis of potential precision oncology approaches, hotspot mutations in HER2, BRAF, KRAS, and PIK3CA were analyzed in 20 surgically resected canine pulmonary adenocarcinomas and three canine pulmonary adenocarcinoma cell lines. HER2 V659E and BRAF V595E mutations were each detected in 3/20 cases (15%), while KRAS G12V was detected in 1/20 cases (5%). No PIK3CA hotspot mutations were identified. The BRAF V595E mutation was additionally detected in the AZACL2 cell line. Functional analysis demonstrated increased sensitivity of AZACL2 cells to the BRAF inhibitor dabrafenib and MEK inhibitors including trametinib, compared with BRAF wild-type cell lines. These findings support MAPK pathway dependency in BRAF-mutant canine pulmonary adenocarcinoma. The mutation spectrum was broadly consistent with previous reports, suggesting a conserved molecular landscape across geographic regions. Collectively, these data identify BRAF and HER2 alterations as clinically relevant candidates for molecular diagnostics and targeted therapy in canine pulmonary adenocarcinoma.

Veterinary Sciences doi: 10.3390/vetsci13060595

Authors: Eduardo Rosales Barahona Andre Luiz Garcia Dias Ashley Egyedy Burim N. Ametaj

Uterine infections (metritis and endometritis) are a leading cause of culling and reproductive failure in transition dairy cows, and antibiotic-resistant Gram-negative pathogens limit conventional therapy. This randomized, controlled, multi-farm field trial evaluated whether four intravaginal infusions of a host-adapted lactic acid bacteria (LAB) cocktail (Lactobacillus sakei FUA3089, Pediococcus acidilactici FUA3138, P. acidilactici FUA3140; 108–109 cfu/dose) at −3, −2, +3, and +4 weeks relative to calving reduce periparturient disease and improve milk production. A total of 526 pregnant cows (426 Holstein, 100 Jersey) from four commercial Alberta farms (automatic-milking, parlor, and certified-organic systems) were block-randomized within farm and parity to TRT1 (saline; n = 175), TRT2 (saline + skim milk; n = 176), or TRT3 (LAB cocktail in saline + skim milk; n = 175). Uterine infection incidence was assessed by Metricheck™ mucus scoring and transrectal ultrasonography at +3 and +4 weeks postpartum. Across the principal peripartum infectious outcomes, TRT3 showed a consistent protective effect: uterine infection incidence was lowest in TRT3 (18.8% vs. 25.1% in pooled controls; OR = 0.69; 95% CI, 0.44–1.09; an approximately 25% relative reduction; exact p = 0.12), and this metritis signal was additionally supported by a repeated-measures mixed model accounting for farm, parity, and week (p = 0.0175), although the Bonferroni-adjusted pairwise contrasts were tendencies (adjusted p ≈ 0.12), and the effect did not differ by parity (treatment × lactation interaction, p = 0.97). Subclinical mastitis was numerically lower in TRT3 than in pooled controls (5.3% vs. 8.9%; OR = 0.57; 95% CI, 0.27–1.24; exact p = 0.16), whereas retained placenta, milk fever, displaced abomasum, and lameness showed no clear cow-level treatment effect in the cow-level exact analyses. Milk yield increased significantly in multiparous cows, which produced 4.6 L/day more milk than TRT1 and 3.22 L/day more than TRT2 over the first 50 days in milk (p < 0.01 for both contrasts; treatment × parity interaction, p = 0.01). No effect was seen on milk composition, uterine involution, or reproductive performance. The trial supports intravaginal LAB as a candidate antibiotic-free prophylactic whose response depends on farm- and cow-level contexts and whose mechanisms require confirmation through microbiological and metabolic measurements.

Veterinary Sciences doi: 10.3390/vetsci13060594

Authors: Zengwen Huang Jing Wang Chaoyun Yang Heng Yang Zhiqiang Hu Gang Shu Zengpeng Lv Dayong Si

Liangshan Yanying chicken is a valuable plateau-adapted indigenous poultry breed in China. The poultry cecum modulates nutrient metabolism, gut microbial colonization and intestinal immune barrier establishment, while the molecular mechanisms driving its age-dependent development during the brooding stage remain unclear. Here, integrated transcriptomic and metabolomic profiling coupled with bioinformatics correlation analysis were conducted on cecal samples collected from chickens at post-hatching days 1, 14 and 28. Significant temporal changes were observed in cecal gene expression and metabolite abundance, and day 14 was identified as a critical window for cecal functional maturation and microbial colonization. In total, 2424 metabolites were annotated, including 600 differentially accumulated metabolites. The cecum exhibited phase-specific metabolic patterns: endogenous energy metabolism dominated at 1–14 d, while lipid biosynthesis prevailed at 14–28 d. The intestinal IgA immune network was verified as the core pathway maintaining cecal immune homeostasis in young chicks. Multi-omics conjoint analysis yielded 53 overlapping KEGG pathways, 14 core pathways, 3 pivotal metabolites and 5 hub genes, based on which three interactive regulatory networks were constructed. Transcriptomic data were validated via qRT-PCR. This study reveals cecal metabolic remodeling and regulatory characteristics during the brooding period, supplementing gut developmental research on plateau indigenous chickens. Notably, these results reflect age-related cecal developmental changes rather than breed-specific high-altitude adaptation mechanisms. Further independent verification is required for metabolomic data and predicted regulatory networks. This finding provides a theoretical basis for scientific breeding and feeding management of Liangshan Yanying chickens.

Veterinary Sciences doi: 10.3390/vetsci13060593

Authors: Yufei Yang Liwei Zeng Ruizi Ren Dongqiang Zheng Yipeng Jin Hao Shi

Dental calculus is a highly prevalent oral condition in dogs and is widely recognized as an important risk factor for gingival inflammation and periodontal disease. Effective strategies for its prevention and treatment remain limited, highlighting the significance of exploring novel mechanisms underlying its formation. Neutrophil extracellular traps (NETs), a key component of innate immunity, have been found in various diseases. To investigate the relationship between NETs and canine dental calculus formation, NET-associated markers were assessed in the oral cavities of dogs with dental calculus and healthy controls. Based on previously published full-length 16S rRNA amplicon sequencing data of canine dental calculus, Porphyromonas gulae was selected as a candidate NET-inducing bacterium for subsequent validation experiments. Subsequent neutrophil stimulation experiments were conducted to explore the effects of NETs and related factors on dental calculus formation. Collectively, our findings demonstrate the presence of NETs within canine dental calculus and reveal that P. gulae present in canine dental calculus is capable of inducing NET formation. The level of myeloperoxidase–DNA complex in gingival crevicular fluid was significantly elevated in dogs with dental calculus. NETs promoted aggregation and microcrystal formation from calcium and phosphate ions under both physiological and supersaturated concentrations. By adhering to the surface of dental calculus, NETs facilitated calculus accumulation. This effect showed positive correlation with neutrophil counts and administration frequency, but was independent of the concentration of administered calcium and phosphate solutions. IL-1β promoted the formation of aggregated NETs but did not enhance calculus accumulation. DNase I inhibited this process by degrading NET-DNA. In conclusion, dental calculus and the calculus-inhabiting P. gulae could stimulate oral neutrophils to release NETs, which participate in and facilitate the initial formation, aggregation, and subsequent accumulation of canine dental calculus.

Veterinary Sciences doi: 10.3390/vetsci13060592

Authors: Qin Gao Chong Yuan Shanglai Li Hua Yang Zongyou Wei Yanli Zhang

Leucine, an essential branched-chain amino acid, serves not only as a substrate for protein synthesis but also as a key regulator of placental function and fetal development. This study investigated the effects of dietary supplementation with RP-Leu during late gestation on placental development and offspring performance in Hu sheep. Sixty twin-pregnant ewes at day 80 of pregnancy were randomly assigned to either a control group (fed a basal diet) or an RP-Leu group (fed a basal diet supplemented with 19 g/day RP-Leu). The feeding trial lasted for 60 d. The ewes were slaughtered at day 140 of gestation. Maternal slaughter traits and fetal organ weights were recorded. Blood and milk samples were collected for milk composition analysis and targeted metabolomic profiling. Leucine supplementation significantly increased the percentage of milk fat content, total solid content, and the birth weight of lambs (p < 0.05). Improvements in placental morphology and antioxidant capacity were observed, including a significant increase in cotyledon density and a significant enhancement of catalase (CAT) activity (p < 0.05). Gene expression analysis indicated that the NOS3, SLC38A1 and FABP4 genes in the placental cotyledons (p < 0.05), and the VEGFA, NOS3, SLC27A1 and FABP4 genes were significantly upregulated in the maternal caruncles (p < 0.05). Plasma metabolomic profiling revealed increased L-glutamic acid levels and alterations in several amino acids, with pathway enrichment indicating involvement in amino acid metabolism and membrane transport processes. Transcriptomic analysis identified 739 differentially expressed genes, which were mainly enriched in the PI3K/Akt signaling pathway, ECM–receptor interaction pathway, and cytokine–cytokine receptor interaction pathway. Collectively, these findings suggest that RP-Leu supplementation during late gestation may enhance offspring growth by modulating amino acid metabolism, promoting placental development, and improving placental nutrient transport capacity, thereby supporting fetal growth and development.

Veterinary Sciences doi: 10.3390/vetsci13060591

Authors: Maureen Kelleher Jacqueline Marr Brittney Graham Thomas Cecere Brett Klamer Sergey Anishchenko David Beylin

Injuries of the metacarpophalangeal joint are a major cause of morbidity and catastrophic fracture in racing horses, yet early osseous pathology is often difficult to detect using conventional imaging. This pilot study aimed to correlate sodium fluoride Positron Emission Tomography (18F-NaF PET) radiopharmaceutical uptake with gross and histopathologic changes in the distal third metacarpal bone (MC3) and proximal first phalanx (P1). Five horses (three racing Thoroughbreds with fetlock injury and two non-racing controls) underwent ante-mortem 18F-NaF PET and cone-beam CT imaging (CBCT), followed by post-mortem gross and histologic examination of predefined anatomic sites. Quantitative PET measures, including maximum standardized uptake value (SUVmax), SUVratio, and PET grade, were compared with gross pathology and histopathologic scores for cartilage and subchondral bone. While there were significant regional correlations between PET metrics and gross or histologic scores at select sites, our results need to be considered in light of the small number of horses evaluated. Correlations between PET metrics and gross pathology score were identified on the distal metacarpus on the lateral dorsal condyle and on proximal P1 for lateral dorsal and mid-P1. Correlation of PET metrics and hyaline cartilage histopathology scores were found for dorsal medial and lateral P1, parasagittal dorsolateral P1 and the medial parasagittal groove of MC3. Correlation of PET metrics and histologic subchondral bone scores were significant for medial palmar condyle, medial parasagittal groove, and parasagittal palmar lateral of MC3. For P1, PET metrics and histologic subchondral bone scores were significantly correlated for parasagittal mid-lateral and medial dorsal regions. Overall, these findings demonstrate that 18F-NaF PET identifies localized bone remodeling that corresponds to histologic and gross pathology at specific fetlock regions, supporting its utility for detecting osseous injury, although relationships varied by anatomic location. Further work with larger numbers of horses is needed.

Veterinary Sciences doi: 10.3390/vetsci13060590

Authors: Lauren R. Finka Ana M. Barcelos James Waterman Avni Bhatia Jenni L. McDonald Rae Foreman-Worsley Beth Skillings

Shelter organisations take responsibility for the care, assessment and homing of large numbers of domestic cats from diverse backgrounds. However, not all cats that come under shelter care are suited to close human-cohabitation or to certain types of human-domestic lifestyles. Shelter stakeholders may undertake decision-making processes to determine how each cat should be managed and where they should go next. These processes may lead to different cat welfare experiences and long-term outcomes depending on how they occur, yet little is known about current approaches. The aim of this study was to characterise current approaches to cat and adopter assessments, behaviour management and homing decisions across the British Isles shelter sector, considering reported practices against sector minimum standards where applicable. A total of 393 quantitative and qualitative responses from employees and volunteers were received. Responses indicated that overall, stakeholders were consistently undertaking cat and prospective adopter assessments, with subsequent information used to support cat management, decision-making and homing. However, the degree of standardisation and objectivity associated with these processes was unclear, with considerable variation in approaches, including certain practices potentially associated with poor cat welfare outcomes identified. Examples include exposure to stressful handling and behavioural interventions and assessment ‘tests’, prolonged stays for harder-to-home cats and potentially suboptimal homing decisions for cats not suited to domestic ‘pet’ lifestyles. Identified opportunities to support welfare-friendly processes at the individual level include more consistent use of cats’ within-shelter behavioural presentations as grounds for ‘pet’-suitability assessments, and careful and consistent application of cat-labelling and terminology. It is also recommended that care is taken to ensure methods of cat assessments, behavioural interventions and homing decisions are pragmatic and optimised to positive cat welfare outcomes.

Veterinary Sciences doi: 10.3390/vetsci13060589

Authors: Katarina Nenadović Dejan Bugarski Tamara Ilić

Background: While traditional pasture-based systems offer sheep natural living conditions and freedom, barn farming provides greater environmental control and protection. The choice of farming system is a crucial factor directly affecting the physical and mental health of the animals. Aim: This study evaluated welfare indicators and parasite prevalence in different sheep management systems, examining the impact of parasitic infections on ewe welfare. Methods: Conducted from November 2025 to April 2026 across 26 farms (13 confined and 13 pasture), the study individually assessed a sample of 1192 ewes aged 2–7 years. Animal-based welfare indicators were assessed using the AWIN protocol for sheep, while parasitic infections were determined from fecal sampling. Results: The results revealed significant differences (p < 0.001) between the systems. Pasture farms showed a higher prevalence of soiled fleece (60.14%), skin lesions, ocular and nasal discharge (6.35%; 31.22%), respiratory issues (16.75%), fecal soiling (16.23%), borderline anemia (16.23%), and excessive itching (16.23%). In contrast, confined systems exhibited higher prevalence of fleece loss (36.00%), hoof overgrowth (20.96%), udder asymmetry (1.44%), and wool pulling (8.32%). Identified parasites included Eimeria spp., gastrointestinal strongyles, Trichuris ovis, Dictyocaulus filarial, Muellerius capillaris, Protostrongylus rufescens, Moniezia spp., Dicrocoelium dendriticum, Fasciola hepatica, Paramphistomum spp., and Giardia intestinalis. Significant correlations (p < 0.001) were found between certain welfare indicators and parasite infections in confined systems, such as between thin body condition score and Eimeria spp., and between fleece quality and fecal soiling with gastrointestinal strongyles, while in pasture systems, thin BCS, nasal discharge, and respiration quality correlated with Muellerius capillaris, borderline anemia with Trichuris ovis and Dictyocaulus filaria, and fecal soiling with Eimeria spp. and Dicrocoelium dendriticum. Conclusions: This data underscores the critical need to improve ewe welfare and implement targeted parasite control strategies in both farming systems.

Veterinary Sciences doi: 10.3390/vetsci13060588

Authors: Yuyang Fan Ge Gao Xinyue Jiang Dongxu Ming Yanpin Li Wenjuan Sun Xilong Li Yu Pi

The objective of this study was to evaluate the effects of partially replacing wheat bran with poplar wood composite fiber (PWCF) on growth performance, immune status, apparent total tract digestibility (ATTD), and gut microbial composition in growing pigs. A total of 140 healthy crossbred (Duroc × Landrace × Yorkshire) growing pigs with an initial body weight of 47.25 ± 0.49 kg were randomly assigned to two dietary treatments, with five replicates per treatment and fourteen pigs per replicate. The control (CT) group was fed a corn–soybean meal-based diet containing wheat bran and rice bran meal, whereas the experimental group received the same diet in which 2% wheat bran was replaced by PWCF. The experiment lasted for 60 days. Compared with the CT group, replacing wheat bran with PWCF did not affect body weight, average daily feed intake, feed conversion ratio, or average daily gain on days 30 or 60 (p > 0.05). In addition, no negative effects were observed on ATTD of nutrients and serum immunoglobulin A (IgA), IgG, and IgM levels at either time point, indicating that PWCF can serve as a suitable partial substitute for wheat bran in growing pig diets. However, it could regulate nitrogen metabolism by reducing blood urea nitrogen (BUN) concentration and the BUN/creatinine ratio, as well as decreasing total free amino acids in serum (p < 0.05). In addition, the antioxidant capacity can be transiently improved by increasing catalase activity. Gut microbiota analysis showed that the replacement significantly increased the relative abundances of Treponema, the Lachnospiraceae_XPB1014_group and Prevotellaceae_UCG-001 (p < 0.05). These changes suggest that PWCF modulates gut microbiota and enriches fiber-degrading bacterial populations. Overall, substituting wheat bran with PWCF did not impair growth performance, immunity, or digestibility, while altering microbial community composition. These findings support the potential application of PWCF as an alternative fiber source, contributing to greater diversity in feed formulation.

Veterinary Sciences doi: 10.3390/vetsci13060587

Authors: Lauren R. Finka Ana M. Barcelos James Waterman Avni Bhatia Jenni L. McDonald Rae Foreman-Worsley Beth Skillings

Meeting the physiological and psychological needs of shelter cats through appropriate care is critical to reducing stress and disease risk, as well as enabling positive homing outcomes. Shelter organisations across the British Isles provide care for many cats; however, little is known about the types of housing and husbandry approaches applied. This study, therefore, aimed to quantify current approaches to cat housing, husbandry, and general care practices, in addition to providing information relevant to local site capacity, considering reported practices against sector minimum standards where applicable. Nine hundred and sixty-one shelter organisations and/or sites caring for cats were identified and invited to complete an online survey including predominantly multiple-choice questions. A total of 393 unique responses were collected from employees and volunteers, and quantitative data were summarised descriptively. In most cases, the results provided evidence of majority alignment with sector standards, although substantial variations in reported practices were also consistently captured. While most responses described approaches supportive of meeting cats’ basic physiological needs (e.g., access to veterinary care and basic resources), psychological needs were addressed less consistently (e.g., general housing and husbandry approaches), potentially leading to poor welfare outcomes. Identified opportunities to better meet cats’ needs include more cat-friendly, low-stress approaches to pen cleaning and cat handling; greater and more consistent provisioning of within-pen resources; and improved approaches to multi-cat housing and associated decision-making. Additional opportunities to enhance both cat and human wellbeing include more structured intake and assessment processes and capacity management to support optimal cat-to-staff ratios, staff working hours, cat lengths of stay and more consistent access to isolation and emergency intake facilities.

Veterinary Sciences doi: 10.3390/vetsci13060586

Authors: Iván Gómez Álvarez José Manuel Verdes García Luciano Espino López

Intervertebral disc disease remains the most common spinal pathology in dogs and is characterized by degeneration of the intervertebral disc, primarily through chondroid metaplasia of the nucleus pulposus. Current histological grading systems for disc degeneration do not include inflammation, despite its potential relevance in clinical presentation through extradural swelling and compression. In this retrospective study, surgically removed disc extrusion material from 74 dogs was histologically processed and evaluated using a semi-quantitative grading system for disc degeneration and inflammation. A high prevalence of chondroid metaplasia was observed, frequently accompanied by inflammatory infiltrates, hemorrhage, and necrosis. A significant positive correlation (correlation coefficient = 0.636, p < 0.001) was found between the degree of degeneration and the intensity of inflammation. Notably, thoracolumbar extrusions exhibited significantly higher inflammatory scores than cervical cases. In Dachshunds, more severe neurological deficits were associated with lower histological degeneration scores, indicating that neurological severity depends on multiple interacting factors that may differ between breeds. No significant associations were identified with age, sex, breed, body weight, neurological localization, or lesion severity in other groups. These findings suggest that inflammation is an active component in the pathophysiology of intervertebral disc disease. Incorporating inflammatory grading into histological grading systems could refine functional outcome predictions and guide therapeutic decisions.

Veterinary Sciences doi: 10.3390/vetsci13060585

Authors: Robert L. Gillette Vijaya V. Indukuri Jeannie Willems Denise Passmore

Rocky Mountain spotted fever (RMSF), caused by Rickettsia rickettsii, is a potentially severe tick-borne disease affecting both dogs and humans. While commercial in-clinic diagnostic tests have improved the detection of several common canine tick-borne diseases, RMSF is not routinely included in many point-of-care screening panels. In addition, infected dogs may present with subtle or nonspecific clinical signs, making diagnosis challenging. This retrospective observational study evaluated the occurrence of PCR-confirmed Rickettsia rickettsii infection in dogs presented to a veterinary lameness and performance referral center. From March 2022 through February 2024, a convenience sample of 51 dogs underwent RMSF testing as part of their diagnostic evaluation. Blood samples were analyzed using a commercial vector-borne disease PCR panel. Overall, 13 of 51 dogs (25.5%) were PCR-positive for Rickettsia rickettsii. Thirty-six dogs originated from North Carolina and South Carolina, of which 13 (36.1%) were PCR-positive. These findings suggest that RMSF should be considered among the differential diagnoses in dogs presenting with unexplained lameness, reduced athletic performance, or other nonspecific clinical concerns in regions where tick-borne diseases are common. The identification of PCR-confirmed Rickettsia rickettsii infection in this referral population highlights the potential value of expanded vector-borne disease testing.

Veterinary Sciences doi: 10.3390/vetsci13060584

Authors: Justyna Kłopotowska Eva Maria Kalbhenn Babette Klein Sabita Diana Stöckle Roswitha Merle Elisabeth Müller Heidrun Gehlen

Bacterial infections in equine wounds are common and can complicate treatment and healing. This five-year retrospective study examined 2844 samples from wounds and abscesses from horses in Germany (2019–2023), analyzing bacterial prevalence by care setting and geographical region. A total of 4464 bacterial isolates were identified, with β-hemolytic streptococci (20.6%), Escherichia coli (15.6%), and Staphylococcus aureus (13.0%) being the most common. Methicillin-resistant Staphylococcus aureus (MRSA) accounted for 35.9% of Staphylococcus aureus isolates. Resistance to third- and fourth-generation cephalosporins was detected in 15.2% of Escherichia coli isolates. Samples from clinics showed a significantly higher MRSA rate, while primary care samples more often contained environmental bacteria. Regional differences in pathogen prevalence and statistically significant temporal changes were observed. The results highlight the importance of continuous microbiological surveillance and underscore the need for targeted antimicrobial stewardship in veterinary facilities. These findings have implications for infection control, empirical treatment strategies, and zoonotic risk mitigation in equine medicine.

Veterinary Sciences doi: 10.3390/vetsci13060583

Authors: Marietta Máté Claire Helen Várnai László Ózsvári

Work-related stress is an important concern in veterinary medicine because it may affect veterinarians’ work, personal life, and well-being. This study described self-reported work-related stressors and their perceived effects on professional and personal life among veterinarians from selected European countries. Between July 2021 and February 2022, an online questionnaire was completed by 724 veterinarians from Hungary, Finland, Sweden, Germany, Denmark, Estonia, and Norway. Participants were recruited through convenience sampling via online channels and professional veterinary networks. The questionnaire assessed 16 stressors, including fatigue, emotional exhaustion, burnout-related symptoms, fear of making mistakes, client expectations, and negative online comments. Mean Likert-scale scores were analyzed using one-way ANOVA and Pearson’s χ2 tests. Fatigue and emotional exhaustion were among the most burdensome internal stressors, with the highest mean score in the Hungarian sample (mean: 4.15 ± 1.05) and the lowest in the Finnish sample (mean: 3.68 ± 1.06; ANOVA: p < 0.0001). Euthanasia-related stress was rated lower in Finland (mean: 1.68 ± 0.83) and Sweden (mean: 1.88 ± 0.95) than in Germany (mean: 2.41 ± 1.17) and Hungary (mean: 2.64 ± 1.27; ANOVA: p < 0.0001). In Hungary, younger and female veterinarians reported greater sensitivity to several stressors. The findings are descriptive and exploratory rather than representative cross-country comparisons.