Veterinary Sciences

10 pages, 793 KB

Open AccessArticle

Development and Validation of a Droplet Digital PCR Assay for Detection of Feline Herpesvirus Type-1

by Yaxi Zhou, Danni Wu, Mengle Tang, Zihan Ye, Erkai Feng, Haili Zhang, Guoliang Luo, Zhenjun Wang, Chunxia Wang, Lina Liu and Yuening Cheng

Vet. Sci. 2025, 12(11), 1107; https://doi.org/10.3390/vetsci12111107 - 20 Nov 2025

Abstract

Feline herpesvirus type-1 (FHV-1), a double-stranded DNA virus, which is a highly infectious upper respiratory tract infection of felids, particularly in kittens. Droplet digital PCR (ddPCR) provides an absolute quantification method with high sensitivity and accuracy. This study aimed to develop a highly [...] Read more.

Feline herpesvirus type-1 (FHV-1), a double-stranded DNA virus, which is a highly infectious upper respiratory tract infection of felids, particularly in kittens. Droplet digital PCR (ddPCR) provides an absolute quantification method with high sensitivity and accuracy. This study aimed to develop a highly sensitive and accurate ddPCR assay for the detection of FHV-1. We designed primers and a probe targeting the FHV-1 glycoprotein D (gD) gene and evaluated the assay’s limit of detection (LOD), sensitivity, repeatability, and specificity in comparison to quantitative real-time PCR (qPCR). The developed ddPCR assay demonstrated a strong linear dynamic range (R² ≥ 0.99) and an exceptionally low LOD of 0.18 copies/μL, which was significantly more sensitive than the method qPCR (LOD ~10 copies/μL). Additionally, the assay exhibited high specificity with no cross-reactivity against other common feline pathogens (feline calicivirus, FCV; feline panleukopenia virus, FPV; feline infectious peritonitis virus, FIPV; Bordetella bronchiseptica and Chlamydia felis) and displayed outstanding repeatability (inter-run CV < 1.35). When applied to 118 clinical samples, the ddPCR assay achieved a significantly higher positive detection rate (27.4%) compared to qPCR (14.8%). In conclusion, we have successfully established a reliable ddPCR assay for the absolute quantification of FHV-1, providing a superior tool for laboratory diagnosis and research. Full article

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18 pages, 278 KB

Open AccessReview

Role of Diet as a Predisposing Factor for Dilated Cardiomyopathy in Dogs: A Narrative Review

by Léa Mornard, Anna Carolina Massara Brasileiro and Mário Marcondes-Santos

Vet. Sci. 2025, 12(11), 1106; https://doi.org/10.3390/vetsci12111106 (registering DOI) - 20 Nov 2025

Abstract

In 2018, the Food and Drug Administration (FDA) reported cases of dilated cardiomyopathy (DCM) in dogs, including breeds without known genetic predisposition, fed diets containing a high proportion of legumes or potatoes, many labeled grain-free. Despite concerns, grain-free diets remain increasingly popular. This [...] Read more.

In 2018, the Food and Drug Administration (FDA) reported cases of dilated cardiomyopathy (DCM) in dogs, including breeds without known genetic predisposition, fed diets containing a high proportion of legumes or potatoes, many labeled grain-free. Despite concerns, grain-free diets remain increasingly popular. This narrative review focuses on the peer-reviewed literature and summarizes recent studies evaluating the effects of grain-free diets on canine cardiac health, with emphasis on taurine deficiency, metabolic alterations, and emerging hypotheses extending beyond nutritional inadequacy. These findings suggest a strong link between diet and DCM, particularly regarding legumes. Dogs of various breeds showed larger left ventricular diameters, reduced systolic function, and increased premature ventricular complexes when fed non-traditional, grain-free, legume-rich diets compared to those on traditional, low-legume diets. Many affected dogs improved clinically and functionally with dietary changes and treatment, suggesting nutritional DCM could be reversible. Though mechanisms remain unclear, the research highlights the potential roles of legumes, particularly peas, in the gut microbiota and fiber-related bile acid metabolism. This review also distinguishes taurine-deficiency-related DCM as a separate form, with certain breeds, notably Golden Retrievers, being more susceptible. Overall, further studies are required to better understand the role of nutrition in canine cardiac health. Full article

16 pages, 630 KB

Open AccessArticle

Genetic Analysis of Stayability and its Relationships with Production, Conformation, Fertility and Health Traits in Holstein Cattle

by Honghong Hu, Zhaodi Xu, Liyun Han, Zhixuan Qiao, Yi Wang, Yikun Jia, Tong Mu and Yun Ma

Vet. Sci. 2025, 12(11), 1105; https://doi.org/10.3390/vetsci12111105 - 19 Nov 2025

Abstract

The objectives of this study were to estimate genetic parameters for stayability in Chinese Holstein cattle and to estimate its genetic correlations with routinely collected traits. Although knowledge and genetic breeding technologies for dairy cattle have advanced, the lifespan of dairy cows has [...] Read more.

The objectives of this study were to estimate genetic parameters for stayability in Chinese Holstein cattle and to estimate its genetic correlations with routinely collected traits. Although knowledge and genetic breeding technologies for dairy cattle have advanced, the lifespan of dairy cows has been declining, mainly due to increased production stress and associated health problems. This makes the genetic improvement of Holstein cattle crucial for sustainable production. Stayability is an indicator trait reflecting longevity, measured as the survival probability of dairy cows at specific stages. To evaluate temporal differences in herd retention, we defined 7 stayability traits as the ability of cows to remain in the herd for 36 (S36), 42 (S42), 48 (S48), 54 (S54), 60 (S60), 72 (S72), and 84 (S84) months after first calving. Data from 56,630 cows between 2011 and 2020 across 14 dairy farms in Ningxia were considered for stayability analyses. Survival analysis using non-parametric methods showed that cattle with sterility had the longest survival time, while those with abomasal displacement had the shortest survival time. The heritability estimates (±standard error) for S36, S42, S48, S54, S60, S72 and S84 were 0.048 ± 0.006, 0.063 ± 0.006, 0.074 ± 0.007, 0.099 ± 0.007, 0.115 ± 0.007, 0.088 ± 0.007, 0.118 ± 0.008, respectively. The genetic and phenotypic correlations among stayability traits ranged from 0.382 to 0.975 and from 0.090 to 0.799, respectively. Furthermore, the approximate genetic correlations between stayability and routinely collected traits (8 production, 20 conformation, 13 fertility, 13 health traits) were assessed. Stayability showed low to moderate genetic correlations with most of these traits. In summary, developing a selection index that incorporates stayability is expected to improve the longevity of dairy cows. All unfavorable genetic relationships observed between stayability and other routinely collected traits should be accounted for in a selection index, thereby enhancing the lifespan of Holstein cattle while maintaining or improving productive performance. Full article

(This article belongs to the Special Issue Advances in Animal Genetics and Sustainable Husbandry)

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13 pages, 4878 KB

Open AccessArticle

SideCow-VSS: A Video Semantic Segmentation Dataset and Benchmark for Intelligent Monitoring of Dairy Cows Health in Smart Ranch Environments

by Lei Yao, Jin Liu, Weinan Hong, Fanrong Kong, Zipei Fan, Lin Lei and Xinwei Li

Vet. Sci. 2025, 12(11), 1104; https://doi.org/10.3390/vetsci12111104 - 19 Nov 2025

Abstract

Accurate and non-invasive monitoring of dairy cows is a cornerstone of precision livestock farming, paving the way for proactive health management and earlier disease detection. The development of robust, AI-driven diagnostic tools, however, is hindered by a dual challenge: scarce realistic video datasets [...] Read more.

Accurate and non-invasive monitoring of dairy cows is a cornerstone of precision livestock farming, paving the way for proactive health management and earlier disease detection. The development of robust, AI-driven diagnostic tools, however, is hindered by a dual challenge: scarce realistic video datasets and a lack of standardized benchmarks for deep learning models. To confront these issues, this study puts forward SideCow-VSS, a video semantic segmentation dataset comprising 921 side-view clips with dense, pixel-level annotations of dairy cows under variable on-farm conditions. We systematically evaluated eight deep learning architectures, from classic convolutional neural networks to state-of-the-art Transformers. The evaluation highlighted a clear performance trade-off: the Mask2Former model with a Swin-L backbone yielded the highest mIoU at 97.32%, making it well-suited for detailed morphological analysis. In contrast, the lightweight PIDNet-s model achieved the fastest inference speed of 59.5 FPS, demonstrating its potential for real-time behavioral alerting systems. This work delivers a foundational resource and quantitative framework to inform model selection, accelerating the creation of computer vision systems for automated health monitoring and adopting preventive strategies against key metabolic and immunological disorders in dairy production. Full article

(This article belongs to the Special Issue Recent Advances in the Diagnosis, Pathogenesis, and Control of Metabolic and Immunological Diseases in Cattle)

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14 pages, 1150 KB

Open AccessArticle

Hematology Reference Values for the Iberian Ribbed Newt (Pleurodeles waltl) Under Human Care

by Carmen Peñas Rodríguez, Manuel Fuertes-Recuero, Teresa Encinas Cerezo, Manuel de la Riva-Fraga, Andrés Montesinos Barceló and Pablo Morón-Elorza

Vet. Sci. 2025, 12(11), 1103; https://doi.org/10.3390/vetsci12111103 - 19 Nov 2025

Abstract

The Iberian ribbed newt (Pleurodeles waltl) is a salamander in the Salamandridae family. Endemic to the Iberian Peninsula and North Africa, it is not commonly found in zoological institutions or wildlife rescue centers. As in other species, routine blood analysis of [...] Read more.

The Iberian ribbed newt (Pleurodeles waltl) is a salamander in the Salamandridae family. Endemic to the Iberian Peninsula and North Africa, it is not commonly found in zoological institutions or wildlife rescue centers. As in other species, routine blood analysis of amphibians under human care is highly recommended, forming an essential component of preventive medicine and effective clinical management. However, despite the great utility of hematological parameters for the diagnosis and prevention of diseases in amphibians, the lack of reliable reference values for many species severely limits their clinical use. The aim of this study is to establish preliminary reference values (RV) for the main hematological parameters in the Iberian ribbed newt. Blood samples were taken from healthy adult individuals (n = 30), females (n = 9) and males (n = 21) maintained under controlled conditions in two zoological institutions. A complete hematological analysis was conducted, which included measurements of hematocrit, total erythrocyte and leukocyte counts, as well as a leukocyte differential. The reference intervals were established according to the guidelines provided by the American Society for Veterinary Clinical Pathology (ASVCP) for sample sizes between 20 and 40 individuals. No significant sex-related differences were detected in the hematological parameters analyzed. Despite the broad reference ranges obtained, these preliminary data provide an essential foundation for the clinical assessment and preventive medical management of P. waltl under human care. Expanding the dataset through collaboration with additional institutions will further refine and improve the accuracy and clinical utility of these reference values. Full article

(This article belongs to the Special Issue Advances in Zoo, Aquatic, and Wild Animal Medicine)

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13 pages, 1807 KB

Open AccessArticle

Quantitative Conspicuity of Pancreatic Canine Insulinoma: A Comparison of Dynamic 4D CT and Dual-Source, Dual-Energy Bolus-Triggered Multiphase CT Imaging

by Veronica Camosci, Claudia Canton, Laura Ventura and Giovanna Bertolini

Vet. Sci. 2025, 12(11), 1102; https://doi.org/10.3390/vetsci12111102 - 19 Nov 2025

Abstract

The aim of this retrospective study was to compare lesion conspicuity—the visibility of a lesion relative to surrounding tissue—and CT characteristics of pancreatic insulinomas in dogs using multiphase dual-energy CT (DECT) and dynamic 4D perfusion CT. Seventy dogs with insulinomas, confirmed either cytologically [...] Read more.

The aim of this retrospective study was to compare lesion conspicuity—the visibility of a lesion relative to surrounding tissue—and CT characteristics of pancreatic insulinomas in dogs using multiphase dual-energy CT (DECT) and dynamic 4D perfusion CT. Seventy dogs with insulinomas, confirmed either cytologically or histologically, or with clinical and imaging findings consistent with the diagnosis, were included. Forty dogs underwent perfusion CT and 30 underwent multiphase DECT on a dual-source, dual-energy CT scanner (192 × 2 detector configuration). Imaging evaluations focused on arterial and portal phases for DECT, and early arterial, late arterial, pancreatic, and portal venous phases for perfusion CT. Tumor conspicuity was quantified using the tumor-to-pancreas ratio (TPR) and contrast-to-noise ratio (CNR), while time-to-peak (TTP) enhancement was recorded for both tumors and pancreatic parenchyma. Perfusion CT demonstrated significantly higher TPR and CNR values compared to DECT (p < 0.001), indicating improved tumor visibility. The late arterial phase of perfusion CT, although not statistically significant, showed the highest median TPR and CNR. Mean TTP for tumors was 38.8 s, slightly earlier than the pancreatic parenchyma (41.25 s). In conclusion, perfusion CT appears to enhance visualization of insulinomas in dogs, particularly between 34 and 44 s after contrast injection, aligning with the late arterial phase of perfusion CT. Full article

(This article belongs to the Special Issue Advances in Small Animal Imaging, Interventional Radiology and Surgical Research)

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15 pages, 1043 KB

Open AccessArticle

Exposure of Wild Ruminants to Toxoplasma gondii in Alpine Ecosystems, NE Spain

by Alejandra Escudero, Maria Puig Ribas, Sonia Almería, Hojjat Gholipour, Lola Pailler-García, Natalia Sastre, Jordi Ruiz-Olmo, Santiago Palazón, Ferran Sayol, Johan Espunyes, Xavier Fernández Aguilar and Oscar Cabezón

Vet. Sci. 2025, 12(11), 1101; https://doi.org/10.3390/vetsci12111101 - 18 Nov 2025

Abstract

Toxoplasma gondii is a protozoan parasite with a complex life cycle that involves warm-blooded animals as intermediate hosts and felids as definitive hosts. Its epidemiology in alpine ecosystems remains poorly understood. This study presents the first long-term investigation of T. gondii exposure in [...] Read more.

Toxoplasma gondii is a protozoan parasite with a complex life cycle that involves warm-blooded animals as intermediate hosts and felids as definitive hosts. Its epidemiology in alpine ecosystems remains poorly understood. This study presents the first long-term investigation of T. gondii exposure in wild ruminants in the Pyrenees (NE Spain), where definitive hosts are scarce. Pyrenean chamois (Rupicapra pyrenaica; n = 1045) and mouflon (Ovis aries musimon; n = 115) sera collected between 2001 and 2024 were tested (Modified Agglutination Test, ELISA-IDvet, ELISA-IDEXX) for the presence of T. gondii antibodies. Sera from 53 chamois and 27 mouflon foetuses and hearts and brains from 38 chamois and 35 mouflon foetuses were analysed for the presence of antibodies and parasite’s DNA, respectively. Moreover, heart and brain (n = 3) and faeces (n = 91) from Pyrenean wildcats (Felis silvestris) were analysed for parasite’s DNA. Seroprevalence was overall low (chamois: 5.24%; mouflon: 1.74%). In multivariate analyses performed in chamois, seroprevalence variation was mainly associated with geographic origin and the diagnostic method used, with little influence of the individual traits of sex or age. No antibodies or T. gondii DNA were detected in chamois or mouflon foetuses. However, chamois did not show a clear age-related increase in exposure to T. gondii, as would be expected if indirect horizontal transmission were the predominant route. In contrast, T. gondii DNA was detected in brain and faecal samples from wildcats, confirming their role as definitive hosts in this ecosystem. Overall, our results reinforce the hypothesis that alpine ecosystems are environments with low T. gondii oocyst contamination and that the parasite most probably relies on sylvatic cycles. These results suggest that wild ruminants, such as chamois and mouflon, could serve as effective sentinels of changes in the extent of this parasite under ongoing environmental and ecological changes. Full article

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20 pages, 1558 KB

Open AccessReview

Swine-Derived Probiotics and Their Metabolites as an Alternative to Veterinary Antibiotics

by Mengshi Zhao, Bihong Chen, Song Peng, Guiheng Mei, Meiqin Li, Fengqiang Lin, Tiecheng Sun and Zhaolong Li

Vet. Sci. 2025, 12(11), 1100; https://doi.org/10.3390/vetsci12111100 - 18 Nov 2025

Abstract

The intensive development of livestock and poultry farming has heavily relied on antibiotics, leading to widespread antimicrobial resistance and posing serious threats to food safety and public health. As the industry transitions towards reduced antibiotic use and sustainable animal production, probiotics and their [...] Read more.

The intensive development of livestock and poultry farming has heavily relied on antibiotics, leading to widespread antimicrobial resistance and posing serious threats to food safety and public health. As the industry transitions towards reduced antibiotic use and sustainable animal production, probiotics and their metabolites have garnered attention as functional alternatives. Probiotics are typically administered in the form of microecological preparations by mixing them into feed or water, offering advantages in cost-effectiveness and ease of use, with demonstrated efficacy in promoting animal health. Swine-derived probiotics, in particular, demonstrate host-specific advantages due to their natural adaptation to the porcine gastrointestinal environment, which improves intestinal colonization, pathogen inhibition, and immune modulation. Their metabolites, including short-chain fatty acids, bacteriocins, and exopolysaccharides, further contribute to these benefits through antimicrobial, anti-inflammatory, and barrier-strengthening effects. Recent studies have demonstrated improvements in average daily gain (18–22%) and feed conversion ratio (12–15%), along with a reduction in diarrhea incidence (up to 40–45%) in weaned piglets supplemented with certain probiotic consortia. It should be noted, however, that part of the supporting evidence is derived from in vitro or non-porcine models, and practical outcomes in swine may vary depending on husbandry conditions, probiotic strain, and husbandry conditions. This review systematically summarizes the isolation and identification of swine-derived probiotics, the active components and functions of their metabolites, and the mechanisms of action and application effects of these metabolites as antibiotic-alternative feed additives. It primarily focuses on innovative research advances in probiotic metabolites for enhancing antibacterial activity and improving pig growth performance. Furthermore, the review discusses the prospects for commercial applications and future research directions, aiming to provide theoretical foundations and technical references for green and healthy farming practices. Full article

(This article belongs to the Special Issue Evaluation of Feeds and Additives on Metabolic or Metabolomic Research on Animal Production)

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18 pages, 993 KB

Open AccessArticle

Comparative Prediction of Methane Production In Vitro Using Multiple Regression Model and Backpropagation Neural Network Based on Cornell Net Carbohydrate and Protein System

by Guanghui Yu, Zenghui Li and Ruilan Dong

Vet. Sci. 2025, 12(11), 1099; https://doi.org/10.3390/vetsci12111099 - 18 Nov 2025

Abstract

Methane (CH₄) produced by methanogenic archaea during the rumen fermentation of feed carbohydrates leads to global warming and total energy loss. This study aims to compare the accuracy of multiple linear regression (MLR) models and backpropagation neural network (BPNN) in predicting [...] Read more.

Methane (CH₄) produced by methanogenic archaea during the rumen fermentation of feed carbohydrates leads to global warming and total energy loss. This study aims to compare the accuracy of multiple linear regression (MLR) models and backpropagation neural network (BPNN) in predicting ruminal CH₄ production of the carbohydrate (Carbs) components of the Cornell Net Carbohydrate and Protein System (CNCPS) in mixed rations of beef cattle with different concentrate-to-forage (C/F) ratios. Two datasets were established using the in vitro fermentation method of Menke and Steingass. One of the datasets contained 60 mixed rations with C/F ratios of 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, and 90:10, respectively, which were used to develop CH₄ prediction models. Another dataset included 10 mixed rations with the same C/F ratios, which were used to validate and compare the accuracy of the prediction models. Results indicated that there was a significant multiple regression relationship between CH₄ production and the Carbs-components (CA (sugars), CB₁ (starch and pectin), CB₂ (available cell wall), CC (unavailable cell wall)) of CNCPS (r² = 0.91, p < 0.0001). An optimal BPNN model with 2 hidden-layer neuron nodes was established with the same variables (r² = 0.93, p < 0.0001). The findings demonstrated that both MLR and BPNN models (p < 0.0001) were suitable for predicting CH₄ production using the Carbs components (CA, CB₁, CB₂, CC) of CNCPS. However, compared with the MLR model, the BPNN model has a greater coefficient of determination (r²) value and concordance correlation coefficient (CCC), and a lower root mean square prediction error (RMSPE), demonstrating better prediction performance. Full article

(This article belongs to the Special Issue Comparative and Functional Anatomy in Veterinary and Animal Sciences)

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14 pages, 738 KB

Open AccessArticle

Evaluating the Role of Social Media in Veterinary Anatomy and Clinical Education: A Student-Based Study

by Ebru Eravci Yalin, Simge Özüner, Zeynep Nilüfer Akçasız, Sevim Güllü and Ozan Gündemir

Vet. Sci. 2025, 12(11), 1098; https://doi.org/10.3390/vetsci12111098 - 18 Nov 2025

Abstract

Social media is increasingly present in higher education, particularly in fields that rely on visual learning such as veterinary anatomy and clinical sciences. This study investigated how veterinary students perceived the use of social media for educational and professional purposes, with specific attention [...] Read more.

Social media is increasingly present in higher education, particularly in fields that rely on visual learning such as veterinary anatomy and clinical sciences. This study investigated how veterinary students perceived the use of social media for educational and professional purposes, with specific attention to its role in learning anatomy and observing clinical procedures. An online cross-sectional survey was administered to students across different academic levels at a veterinary faculty. The questionnaire included demographic questions and eight Likert-scale items addressing the educational value of social media, its role in accessing learning materials, engagement with professional visuals, and ethical concerns regarding the sharing of clinical or surgical content. Data were analyzed using one-way ANOVA and post hoc Tukey tests to assess differences by academic year, age group, and daily social media usage. Analysis showed that students in earlier academic years generally held more favorable views on the use of social media for accessing learning materials and understanding complex subjects. Students in advanced years expressed greater ethical concern, particularly about sharing surgical or clinical videos online. Daily social media usage showed limited influence on general perceptions, though some variation appeared in topic-specific responses. The results suggest that veterinary programs may benefit from structured educational components on digital professionalism and responsible media use to better align student engagement with ethical standards and educational objectives. Full article

(This article belongs to the Special Issue Advances in Veterinary Anatomy: Connecting Basic Research with Clinical, Biotechnological, and Educational Innovations)

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11 pages, 320 KB

Open AccessArticle

Assessment of Bicarbonate Deficiency in Feline Acute and Chronic Kidney Disease

by Francesca Perondi, Matilde Vernaccini, Silvia Morelli, Veronica Marchetti and Ilaria Lippi

Vet. Sci. 2025, 12(11), 1097; https://doi.org/10.3390/vetsci12111097 - 18 Nov 2025

Abstract

Bicarbonate deficiency is a common complication of acute kidney injury (AKI) and chronic kidney disease (CKD), resulting from impaired renal bicarbonate handling. It promotes bone demineralization and calcium–phosphate (CaxP) imbalance. In dogs, elevated CaxP has been linked to increased frequency and severity of [...] Read more.

Bicarbonate deficiency is a common complication of acute kidney injury (AKI) and chronic kidney disease (CKD), resulting from impaired renal bicarbonate handling. It promotes bone demineralization and calcium–phosphate (CaxP) imbalance. In dogs, elevated CaxP has been linked to increased frequency and severity of bicarbonate deficiency. This 10-year study evaluated the prevalence and degree of bicarbonate deficiency in 618 cats diagnosed with AKI, acute-on-chronic kidney disease (ACKD), and CKD, and its association with azotemia and mineral imbalance. A retrospective analysis was conducted on serum biochemical profiles, including creatinine, urea, ionized and total calcium, phosphate, CaxP, and bicarbonate. Cats with incomplete datasets or receiving sodium bicarbonate therapy were excluded. According to IRIS guidelines, bicarbonate deficiency was defined as <16 mmol/L and categorized as moderate (12–16 mmol/L) or severe (<12 mmol/L). Deficiency was found in 45% of cats, of which 63% were moderate and 37% severe. Prevalence was higher in AKI (58%) and ACKD (60%) compared to CKD (38%) (p = 0.002). Negative correlations between bicarbonate and creatinine, phosphate, and CaxP were detected in AKI and CKD, while in ACKD, only creatinine and phosphate correlated. Deficiency increased with disease severity in AKI (p = 0.0145) and CKD (p < 0.0001). Cats with CaxP > 70 mg²/dL² had higher deficiency rates (p < 0.0001). However, 21% with elevated CaxP had normal bicarbonate, suggesting mineral imbalance may occur independently of acidosis. Full article

(This article belongs to the Special Issue Advances in Veterinary Nephrology and Urology of Small Animals)

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15 pages, 675 KB

Open AccessArticle

Radiofrequency Wave Sensing for Rapid Animal Health Monitoring: A Proof-of-Concept Study

by Aftab Siddique, Ramya Kota, Goutham Kumar Isai, Davia Brown, Oreta Samples, Niki Whitley, Phaneendra Batchu, Thomas H. Terrill and Jan van Wyk

Vet. Sci. 2025, 12(11), 1096; https://doi.org/10.3390/vetsci12111096 - 18 Nov 2025

Abstract

Anemia caused by gastrointestinal parasitism is a major constraint to small ruminant productivity, particularly in low-resource production systems where diagnostic tools and veterinary access are limited, with use of FAMACHA as a biological reference This study evaluated the potential of radio-frequency non-destructive technique [...] Read more.

Anemia caused by gastrointestinal parasitism is a major constraint to small ruminant productivity, particularly in low-resource production systems where diagnostic tools and veterinary access are limited, with use of FAMACHA as a biological reference This study evaluated the potential of radio-frequency non-destructive technique (RF-NDT) wave-derived features as non-invasive biomarkers for anemia detection in goats, using FAMACHA© scores as a biological reference. Variable clustering of the top ten frequencies revealed distinct patterns across health states. Healthy (FAMACHA© 1) animals were characterized by a single frequency cluster centered at 8.43 GHz, which explained 93.7% of variation, whereas moderately affected animals (FAMACHA© 2) shifted to 9.33 GHz with reduced uniformity (88.7%). Borderline animals (FAMACHA© 3) required two clusters (9.89 and 8.23 GHz), explaining 91.0% of variation, indicating increasing tissue heterogeneity with anemia progression. Regression analysis demonstrated strong predictive power, with Linear Regression achieving R² = 1.00 and Random Forest R² = 0.79 (RMSE = 0.07), Support Vector Regression underperformed (R² = 0.31). Classification models confirmed the feasibility of categorical anemia detection. The Multilayer Perceptron achieved the highest accuracy (0.84), F1-score (0.83), and ROC-AUC (0.94), outperforming Support Vector Machine (accuracy 0.67, F1 = 0.67) and K-Nearest Neighbors (accuracy 0.60, F1 = 0.61). These findings establish proof-of-concept that RF waves capture physiologically meaningful dielectric signatures linked to anemia, reflecting hemoglobin concentration, hydration, and microcirculatory function. The integration of RF sensing with machine learning offers a rapid, and non-invasive scalable diagnostic approach. Future work should expand validation across breeds and environments, optimize sensor design, and embed neural classifiers for field-ready deployment. Full article

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23 pages, 519 KB

Open AccessReview

A Comprehensive Review: Molecular Diagnostics and Multi-Omics Approaches to Understanding Bovine Respiratory Disease

by Stephanie O’Donoghue, Sinéad M. Waters, Derek W. Morris and Bernadette Earley

Vet. Sci. 2025, 12(11), 1095; https://doi.org/10.3390/vetsci12111095 - 17 Nov 2025

Abstract

Bovine respiratory disease (BRD) is a multifactorial syndrome and a leading cause of morbidity and economic loss in global cattle production. Next-generation sequencing (NGS) platforms, including Illumina and Oxford Nanopore Technologies (ONT), have enabled high-resolution profiling of the bovine respiratory microbiome and virome, [...] Read more.

Bovine respiratory disease (BRD) is a multifactorial syndrome and a leading cause of morbidity and economic loss in global cattle production. Next-generation sequencing (NGS) platforms, including Illumina and Oxford Nanopore Technologies (ONT), have enabled high-resolution profiling of the bovine respiratory microbiome and virome, revealing novel viral contributors such as bovine rhinitis A virus (BRAV) and influenza D virus (IDV). Transcriptomic approaches, including RNA sequencing (RNA-Seq) and microRNA (miRNA) profiling, provide insights into host immune responses and identify potential biomarkers for disease prediction. Traditional diagnostic methods—culture, ELISA, and immunohistochemistry—are increasingly complemented by PCR-based and metagenomic techniques, improving sensitivity and specificity. Despite technological progress, gaps remain in virome characterization, miRNA function, and the integration of multi-omics data. Standardized protocols and longitudinal studies are needed to validate microbial signatures and support field-deployable diagnostics. Advances in bioinformatics, particularly network-based integrative pipelines, are becoming essential for harmonizing multi-omics datasets and revealing complex host–pathogen interactions. The objective of this comprehensive review was to synthesize current understanding of the bovine transcriptomic response to BRD as well as the respiratory microbiome and virome, emphasizing how advanced sequencing technologies have transformed microbial profiling and molecular diagnostics in BRD. Full article

(This article belongs to the Section Anatomy, Histology and Pathology)

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14 pages, 2829 KB

Open AccessArticle

A Hydroxynaphthol Blue-Based Loop-Mediated Isothermal Amplification Assay for Closed-Tube Detection of the Streptomycin Resistance Gene aadA1 in Salmonella

by Yuxiang Shen, Yeqing Zheng, Meiquan Li, Yanli Du, Heng Yang, Fangjie Li, Bin Wang and Xiao Wang

Vet. Sci. 2025, 12(11), 1094; https://doi.org/10.3390/vetsci12111094 - 17 Nov 2025

Abstract

The aadA1 gene, which confers resistance to streptomycin, is typically located within class Ⅰ integrons. This genetic context enables its dissemination among diverse Gram-negative bacteria, such as Salmonella, and facilitates its potential transfer to humans through the food chain or into agricultural [...] Read more.

The aadA1 gene, which confers resistance to streptomycin, is typically located within class Ⅰ integrons. This genetic context enables its dissemination among diverse Gram-negative bacteria, such as Salmonella, and facilitates its potential transfer to humans through the food chain or into agricultural environments via manure. Hence, the detection of aadA1 genes is crucial for surveillance, understanding transmission dynamics, and informing strategies to mitigate the spread of resistant bacteria. Conventional aadA1 detection relies on time-consuming or equipment-intensive molecular assays like PCR or qPCR. In this study, we developed and optimized a closed-tube, hydroxynaphthol blue (HNB) -based loop-mediated isothermal amplification (LAMP) assay to detect Salmonella aadA1 gene and performed evaluation and validation against conventional PCR. The LAMP assay demonstrated high specificity and sensitivity, with a detection limit of 1 pg (190 copies) of genomic DNA per reaction, which is tenfold higher than that of conventional PCR. In parallel testing of 40 Salmonella DNA samples, the optimized LAMP assay achieved a detection rate of 100.0% for the aadA1 gene in streptomycin-resistant isolates, compared to 96.3% by conventional PCR. Among the streptomycin-susceptible isolates, the LAMP assay also showed a higher detection rate (38.5%) for the aadA1 gene, compared to 23.1% by conventional PCR. Consequently, the LAMP assay developed in this study for detecting the aadA1 gene offers a combination of simplicity, speed, visual readout, high specificity, and sensitivity, making it particularly suitable for rapid field detection in antimicrobial resistance surveillance programs. Full article

(This article belongs to the Section Veterinary Microbiology, Parasitology and Immunology)

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25 pages, 3465 KB

Open AccessArticle

Effects of Bacteroides fragilis and Enterococcus faecium Administration as Probiotic Candidates: Impact on Growth Performance, Organ Indices, and Gut Microbiota Balance in Mice

by Mohamed Osman Abdalrahem Essa, Cheng Cheng, Liang Chen, Geng-Yu Chi, Layla Ahmed Mohammed Abdelhadi, Huda Ahmed Hassan, Saniya Yaqoob, Saber Y. Adam, Hosameldeen Mohamed Husien, Ahmed A. Saleh and Darong Cheng

Vet. Sci. 2025, 12(11), 1093; https://doi.org/10.3390/vetsci12111093 - 17 Nov 2025

Abstract

This research was conducted to evaluate the safety and effects of two probiotic candidate strains, Enterococcus faecium (E. faecium) and B. fragilis (BF), which were isolated from healthy sheep, on various factors such as intestinal structure, growth performance, organ [...] Read more.

This research was conducted to evaluate the safety and effects of two probiotic candidate strains, Enterococcus faecium (E. faecium) and B. fragilis (BF), which were isolated from healthy sheep, on various factors such as intestinal structure, growth performance, organ indices, and gut microbiota in female C57BL/6J mice. A total of fifty mice were categorized into five groups: BF108, BF109, EF108, EF109, and a control group. The results indicated that mice in the EF108 and BF109 groups experienced a significant increase in body weight by day 28 compared to the control group (p < 0.05). Additionally, the liver index in the BF108 and EF108 groups was significantly lower (p < 0.05) than that of the control group. The length of the colon was also significantly greater (p < 0.05) in the EF109 and BF109 groups relative to the control group. According to the linear discriminant analysis effect size (LEfSe), the presence of probiotics led to an increased relative abundance of Pediococcus acidilactici in the EF109 and BF109 groups, while Lactiplantibacillus plantarum was the most prevalent strain in the EF108 and BF108 groups. Finally, Spearman correlation analysis demonstrated a positive association between Bacillota and Bacteroidota as well as Verrucomicrobiota. Additionally, Bacteroidota displayed positive relationships with Verrucomicrobiota and Planctomycetota, while showing a negative correlation with Actinomycetota and Gemmatimonadota, supporting its potential use as next-generation probiotics. In summary, our results indicate probiotics may have a role in promoting intestinal health by enhancing intestinal shape and histological characteristics. Full article

(This article belongs to the Special Issue Intestinal Homeostasis and Its Regulatory Role in Animal Growth and Development)

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12 pages, 543 KB

Open AccessArticle

Pregnancy-Associated Glycoproteins Identification in Skopelos Goat Milk by Means of Mass Spectrometry

by Efterpi Bouroutzika, Ekaterini K. Theodosiadou, Stavros Proikakis, Irene Valasi and George Th. Tsangaris

Vet. Sci. 2025, 12(11), 1092; https://doi.org/10.3390/vetsci12111092 - 17 Nov 2025

Abstract

The proteins most frequently associated with pregnancy are the pregnancy-associated glycoproteins (PAGs), which are abundantly expressed placental products in species belonging to the order Cetartiodactyla. Multiple PAG isoforms are expressed across different species and stages of gestation, with their expression influenced by factors, [...] Read more.

The proteins most frequently associated with pregnancy are the pregnancy-associated glycoproteins (PAGs), which are abundantly expressed placental products in species belonging to the order Cetartiodactyla. Multiple PAG isoforms are expressed across different species and stages of gestation, with their expression influenced by factors, such as breed and bodyweight. These proteins can be detected in both blood and milk samples using ELISA or RIA assays, serving as early indicators of pregnancy. The present study aimed to detect PAGs in caprine milk through a non-invasive, high-throughput, mass-spectrometry-based proteomic approach. Milk samples were collected from 20 Skopelos breed goats at 20 and 45 days post-mating. Following appropriate processing for whey protein extraction, the samples underwent tryptic digestion to generate peptides for LC-MS/MS analysis. The proteomic investigation identified five distinct PAGs (caPAG₂ on day 20 and caPAG₃, caPAG₅, caPAG₆, and caPAG₁₂,on day 45 post-mating) annotated to Capra hircus, along with 22 additional proteins associated with the fetal–maternal interface, pregnancy progression, and immune-related pathways. These findings demonstrate that LC-MS/MS provides a non-invasive, sensitive, and reliable method for detecting PAGs in caprine milk during the early stages of pregnancy (day 20, as the presence of caPAG₂ revealed), applicable to both individual animals and flock-level monitoring. Full article

(This article belongs to the Section Veterinary Reproduction and Obstetrics)

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17 pages, 1831 KB

Open AccessArticle

Evaluation of a Multimodal Anesthetic Protocol for Immobilization in Black Vultures (Coragyps atratus) and Turkey Vultures (Cathartes aura)

by Alejandro Vargas Araya, Jeff C. Ko, Tomohito Inoue, Shane Guenin, Tyler C. Hunt, Patrice E. Baumhardt and Esteban Fernández-Juricic

Vet. Sci. 2025, 12(11), 1091; https://doi.org/10.3390/vetsci12111091 - 16 Nov 2025

Abstract

Anesthetic protocols for non-invasive immobilization of Black Vultures (Coragyps atratus) and Turkey Vultures (Cathartes aura) for procedures lasting up to two hours are lacking. This study directly evaluated the safety and efficacy of a multimodal anesthetic protocol in 11 [...] Read more.

Anesthetic protocols for non-invasive immobilization of Black Vultures (Coragyps atratus) and Turkey Vultures (Cathartes aura) for procedures lasting up to two hours are lacking. This study directly evaluated the safety and efficacy of a multimodal anesthetic protocol in 11 Black Vultures and 4 Turkey Vultures undergoing electroretinography (ERG). Vultures were anesthetized with intramuscular dexmedetomidine (5 μg/kg), midazolam (0.2 mg/kg), butorphanol (0.2 mg/kg), and ketamine (5 mg/kg) (DMBK), followed by isoflurane induction and maintenance. All vultures were mechanically ventilated to maintain consistent end-tidal CO₂. Monitored parameters included sedation and recovery quality, heart and respiratory rates, hemoglobin oxygen saturation, non-invasive blood pressure, body temperature, and end-tidal concentrations of CO₂, isoflurane, and oxygen. All vultures achieved profound sedation with smooth induction and a median isoflurane maintenance concentration of 1.4% for approximately two hours. Recovery was rapid and uneventful. Heart rates ranged from 60 to 119 beats/min. Mean arterial blood pressure averaged 149 mmHg in Black Vultures and 158 mmHg in Turkey Vultures, with Turkey Vultures showing significantly higher diastolic pressure. A second-degree heart block was detected in one Black Vulture but required no treatment. All ERG procedures were completed successfully. The DMBK protocol provided profound sedation in both species, maintained key cardiorespiratory parameters, including heart rate and arterial blood pressure, within a clinically acceptable range throughout isoflurane maintenance, and enabled uneventful rapid recovery. These findings support DMBK with the cardiorespiratory monitoring system as a safe and effective regimen for anesthetizing vultures and likely other similarly sized raptors requiring non-invasive immobilization. Full article

(This article belongs to the Section Veterinary Surgery)

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9 pages, 743 KB

Open AccessBrief Report

The Concentration and Duration of Lipopolysaccharide Stimulation Produce Different Cytokine Responses in an Ex Vivo Whole Blood Model in Horses

by Natalie Mitlyng, Kallie J. Hobbs, Bethanie L. Cooper and M. Katie Sheats

Vet. Sci. 2025, 12(11), 1090; https://doi.org/10.3390/vetsci12111090 - 16 Nov 2025

Abstract

Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited [...] Read more.

Lipopolysaccharide (LPS) is frequently used in equine research to model clinical endotoxemia; however, there is no standardized protocol for inducing cytokine production in equine whole blood. To address this knowledge gap, the goal of this study was to compare the cytokine responses elicited by three different LPS stimulation protocols. Whole blood was collected from six healthy horses (aged 5–30 years; mixed breeds and genders) from the North Carolina State University teaching herd (IACUC #23-412). Sixty milliliters of heparinized blood were aseptically drawn and divided into 15 mL aliquots. Samples were stimulated with LPS at concentrations of 100 ng/mL, 1000 ng/mL, or using a two-hit model (500 ng/mL initially and again at 1.5 h). Incubation occurred at 37 °C on an orbital shaker for time points ranging from 1.5 h to 24 h. Cytokine concentrations were measured using the Cornell Equine Cytokine and Chemokine Panel and were compared to non-stimulated controls. LPS stimulation induced the production of TNF-α, IL-1β, IL-10, CCL5, and CCL11 in a time- and concentration-dependent manner. Notably, reliable and robust cytokine responses were observed only after 12 h of stimulation with either 1000 ng/mL or the two-hit 500/500 ng/mL protocol. These findings suggest that both the concentration and duration of LPS exposure significantly influence cytokine expression in equine whole blood. Therefore, the optimal stimulation protocol may vary depending on the specific cytokine of interest, and careful consideration of these variables is essential for designing reproducible and physiologically relevant ex vivo models of equine endotoxemia. Full article

(This article belongs to the Topic Application of Animal Models: From Physiology to Pathology)

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15 pages, 435 KB

Open AccessArticle

Effects of Lidocaine Alone Versus Lidocaine–Dexmedetomidine Infusion on Pulmonary Gas Exchange and Respiratory Mechanics During Isoflurane Anesthesia in Horses

by Ludovica Chiavaccini, Raiane A. Moura, Tatiana Moreira Batista P. R. Azevedo, Chiara De Gennaro, Enzo Vettorato, Marta Romano and Diego A. Portela

Vet. Sci. 2025, 12(11), 1089; https://doi.org/10.3390/vetsci12111089 - 16 Nov 2025

Abstract

Dexmedetomidine improves pulmonary function in dogs and humans, but evidence in horses is scarce. This study evaluated dexmedetomidine infusion on oxygenation and respiratory mechanics in anesthetized horses. Twenty horses undergoing elective surgery were included in a prospective, non-randomized, observational study. Horses received either [...] Read more.

Dexmedetomidine improves pulmonary function in dogs and humans, but evidence in horses is scarce. This study evaluated dexmedetomidine infusion on oxygenation and respiratory mechanics in anesthetized horses. Twenty horses undergoing elective surgery were included in a prospective, non-randomized, observational study. Horses received either lidocaine alone (1.3 mg/kg over 15 min, then 3 mg/kg/hour; LIDO) or combined with dexmedetomidine (1.75 μg/kg over 15 min, then 1.75 μg/kg/hour; DL). Respiratory mechanics, gas exchange, and cardiovascular variables were recorded at baseline, post-loading, and after 30, 60, and 90 min. Data were analyzed using mixed-effects linear models with horse as a random effect and time and treatment and their interaction as fixed effects (p ≤ 0.05). Peak inspiratory pressure increased over time with both treatments but was lower with DL at 90 min (−1.26 mmHg, p = 0.046). There was no evidence that arterial oxygen pressure or oxygenation ratio improved over time with DL (p > 0.75). Shunt fraction did not significantly change over time or between treatments (Wald χ² = 4.77, p = 0.85). Heart rate with DL decreased from baseline (p ≤ 0.001) but remained higher than LIDO overall (p = 0.001). Dexmedetomidine infusion showed no benefit on oxygenation or respiratory mechanics in anesthetized horses. Full article

(This article belongs to the Special Issue Emerging Trends in Veterinary Anesthesia and Analgesia)

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