Animals

13 pages, 3008 KB

Open AccessArticle

Prevalence and Genomic Features of Aichivirus D in Yaks in Western China

by Yang Su, Musha Jili, Guangfu Zhao, Qibing Gu, Falong Yang, Hui Zhang, Huanrong Zhang, Dechun Chen, Yutao Shi, Mingjing Gu, Kebu Munai, Wangqing Banma and Kehamo Abi

Animals 2026, 16(18), 2902; https://doi.org/10.3390/ani16182902 (registering DOI) - 15 Sep 2026

Abstract

Aichivirus D (AiV-D), a member of the genus Kobuvirus within the family Picornaviridae, is an emerging pathogen associated with diarrhea in multiple livestock species, including cattle, dairy cattle, Tibetan sheep, and yaks. In this study, a total of 280 fecal samples were collected [...] Read more.

Aichivirus D (AiV-D), a member of the genus Kobuvirus within the family Picornaviridae, is an emerging pathogen associated with diarrhea in multiple livestock species, including cattle, dairy cattle, Tibetan sheep, and yaks. In this study, a total of 280 fecal samples were collected from yaks with diarrhea in Sichuan, Gansu, and Qinghai Provinces, western China, between June 2025 and February 2026. The overall positive rate of AiV-D was 30.00% (84/280), and the farm-level positive rate was 87.5% (7/8). Four complete yak AiV-D genomes (GenBank: PZ424445–PZ424448) were obtained from the positive samples, with genome sizes ranging from 8369 to 8475 nt and GC contents ranging from 55.66 to 55.85%. Each genome contained a single open reading frame (ORF) of 7509–7518 bp, encoding a typical picornaviral polyprotein consisting of a leader protein (L), structural proteins P1 (VP0, VP3, and VP1), and nonstructural proteins P2 (2A, 2B, and 2C) and P3 (3A, 3B, 3C, and 3D). Pairwise comparisons of the ORF revealed nucleotide and amino acid identities of 82.07–99.63% and 81.96–99.56%, respectively, among the four strains. Phylogenetic analysis based on the complete genomes revealed that strain BKV/YAK/FX1/2025/CHN clustered with the bovine reference strain BKV5 (84.70% nt/88.35% aa identity), whereas the other three strains grouped with the yak reference strain BKV/Yak/SCDF77/2021/CHN (GenBank: OP776109.1) (89.80–93.07% nt 95.53–96.84% aa identity). Recombination analysis predicted a recombination event in strain BKV/YAK/SF1JK6/2025/CHN, with breakpoints located in the nonstructural protein coding region (2C–3D). Additionally, eight full-length VP0, VP3, and VP1 gene sequences were amplified from the AiV-D-positive samples, and a few co-mutational amino acid substitutions were identified in these structural proteins. Phylogenetic analysis based on the VP1 gene further divided the eight strains into two distinct clades: six strains formed an independent clade and were designated as a novel sub-genotype within AiV-D4, tentatively named AiV-D4-L1, closely related to but clearly distinguishable from the AiV-D4 reference strains reported in China, while the remaining two strains (BKV/YAK/FX1/2025/CHN and BKV/YAK/FX77/2025/CHN) clustered with the bovine reference strain and were designated as a novel genotype, AiV-D5. These results reveal genetic diversity among yak-derived AiV-D strains and suggest possible host adaptability or cross-host transmission, warranting further investigation in future studies. However, as all the samples were collected from diarrheic yaks without a healthy control group, the potential association between AiV-D and diarrhea requires further validation. Full article

(This article belongs to the Section Small Ruminants)

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42 pages, 2059 KB

Open AccessReview

Preparation of Targeted Delivery Materials and Their Application in Animal Production

by Bingfeng Zheng, Yingcheng Gao, Kaisi Hu, Wei Zhang, Wenjie Zhang and Jian Ma

Animals 2026, 16(18), 2901; https://doi.org/10.3390/ani16182901 (registering DOI) - 15 Sep 2026

Abstract

In livestock production, conventional nutrients and bioactive substances are susceptible to degradation during feed processing, storage, and gastrointestinal transit, resulting in low bioavailability and feed conversion efficiency. Targeted delivery technology, which utilizes carriers such as nanoparticles, microcapsules, and hydrogels, can protect these compounds [...] Read more.

In livestock production, conventional nutrients and bioactive substances are susceptible to degradation during feed processing, storage, and gastrointestinal transit, resulting in low bioavailability and feed conversion efficiency. Targeted delivery technology, which utilizes carriers such as nanoparticles, microcapsules, and hydrogels, can protect these compounds from premature degradation, regulate their release kinetics, and achieve site-specific accumulation via various administration routes, including oral and injectable delivery. This paper introduces the main types of targeted delivery materials and their preparation methods: ionotropic gelation, emulsion solvent evaporation, and nanoprecipitation for nanoparticle fabrication; spray drying, orifice coagulation bath, ionic crosslinking, complex coacervation, and electrospinning for microcapsule production; and ionic crosslinking, gas shear-assisted ionic crosslinking, oil-in-water emulsion combined with calcium ion crosslinking, and enzymatic crosslinking for gel-based system synthesis. Evaluation methods are also described in detail, covering particle size and polydispersity index analysis, morphological characterization, in vitro simulated digestion, and in vivo fluorescence imaging. This paper reviews the applications of this technology in poultry, pigs, and ruminants, with emphasis on its roles in enhancing intestinal health, regulating immune responses, inhibiting pathogenic bacteria, and improving growth performance and product quality, aiming to provide theoretical references for the rational design and application of targeted delivery systems in precision animal nutrition and sustainable livestock production. Full article

(This article belongs to the Special Issue Innovations in Sustainable Animal Nutrition: From Feed Evaluation to Precision Feeding)

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19 pages, 2071 KB

Open AccessArticle

Biological Load and Tray Position Shape Larval Yield and Frass Production in a Stacked Hermetia illucens Rearing System

by Alondra Guerrero-Hernández, Aurelio Guevara-Escobar, Juan Fernando García-Trejo, Ximena Galilea Ruiz-Maldonado and Ana Angelica Feregrino-Pérez

Animals 2026, 16(18), 2900; https://doi.org/10.3390/ani16182900 (registering DOI) - 15 Sep 2026

Abstract

Industrial black soldier fly (Hermetia illucens) production commonly relies on stacked tray systems, yet little is known about how tray position and biological load influence production heterogeneity. This study evaluated whether different initial biological loads, imposed by varying the initial larval [...] Read more.

Industrial black soldier fly (Hermetia illucens) production commonly relies on stacked tray systems, yet little is known about how tray position and biological load influence production heterogeneity. This study evaluated whether different initial biological loads, imposed by varying the initial larval mass while maintaining a constant numerical density, together with tray position, influence larval biomass and dry frass production. Same-age third-instar larvae were seeded at a constant density of 9000 larvae per tray under five biological-load treatments (29, 54, 64, 97 and 200 g tray⁻¹, fresh weight basis). Initial biological load affected wet larval biomass, dry larval biomass, and dry frass production. However, higher initial biological load did not produce proportional gains in final biomass. The lowest initial biological load resulted in the greatest final wet larval biomass (3.54 kg tray⁻¹) whereas the highest initial biological load resulted in only 1.75 kg tray⁻¹ (

p = 0.025

). Intermediate tray positions produced significantly greater wet larval biomass than the bottom tray. Mean substrate temperature did not differ among tray positions, whereas moisture declined over time and varied by position. These findings demonstrate that production consistency in stacked H. illucens rearing systems depends not only on biological load but also on tray-level microenvironmental heterogeneity, providing practical guidance for the design and management of industrial stacked-rearing systems. Full article

(This article belongs to the Section Animal System and Management)

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15 pages, 764 KB

Open AccessArticle

Feeding Pigs Without Inorganic Phosphate Supplementation from Weaning to Slaughter: Contribution to More Sustainable Swine Production

by Jakob Leskovec, Joan Tarradas, Núria Tous, Beatriz Jimenez-Moya, Marc Bagaria, Peter Ader and David Torrallardona

Animals 2026, 16(18), 2899; https://doi.org/10.3390/ani16182899 - 15 Sep 2026

Abstract

The viability of raising pigs from weaning to slaughter with inorganic phosphorus (iP)-free diets using a hybrid 6-phytase was tested in two trials. Three experimental diets were formulated: T1, a control diet with monocalcium phosphate (MCP); T2, a diet without MCP + phytase [...] Read more.

The viability of raising pigs from weaning to slaughter with inorganic phosphorus (iP)-free diets using a hybrid 6-phytase was tested in two trials. Three experimental diets were formulated: T1, a control diet with monocalcium phosphate (MCP); T2, a diet without MCP + phytase (P and Ca phytase matrix formulation); and T3, a diet without MCP + phytase (P, Ca + extra nutrients in a phytase matrix formulation). Rapeseed and sunflower meal were used in addition to soybean meal as protein sources, in order to increase the phytate-P content. Performance was monitored from weaning to slaughter, and bones were sampled at the slaughterhouse to test for bone mineralization. In the second trial, gait scores were also assessed at 75, 110, and 134 days. Weight gain was not significantly affected by treatment in either trial. However, in Trial 1, males receiving T2 had a reduced overall feed efficiency than males receiving T1, whereas in Trial 2, a reduced overall feed efficiency was observed for T3 relative to T2 (p < 0.05), probably due to the extra nutrient reduction. No differences in bone mineralization or gait score were observed. In conclusion, it is possible to raise pigs without iP by using phytase in feeds containing phytate-P. This may contribute to more sustainable pig production with reduced P excretion and lower usage of rock phosphate, a limited resource. Full article

(This article belongs to the Special Issue Modification of Pig Feeding as a Need for Sustainable Agriculture: Strengths and Weaknesses of Such Activities)

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12 pages, 233 KB

Open AccessArticle

Enteric Methane Emission and Milk Component Profiles of Holstein and Jersey Cows: A Respiration Chamber Study

by Jeongkuk Park, Jun Sik Eom, Dong Hyeon Kim, Seong Min Park, Su Bin Han, Younglae Kim, Chang-Gwon Dang and Dong-Hyun Lim

Animals 2026, 16(18), 2898; https://doi.org/10.3390/ani16182898 - 15 Sep 2026

Abstract

Holstein and Jersey dairy cows differ in body size, feed intake, milk production, and milk composition, which may influence the interpretation of enteric methane (CH₄) emissions. This study compared production traits, milk composition, and respiration chamber-measured CH₄ emissions between 10 [...] Read more.

Holstein and Jersey dairy cows differ in body size, feed intake, milk production, and milk composition, which may influence the interpretation of enteric methane (CH₄) emissions. This study compared production traits, milk composition, and respiration chamber-measured CH₄ emissions between 10 Holstein and 10 Jersey cows. Repeated measurements were averaged within cow, and breed differences were evaluated using independent two-sample t-tests. Sensitivity analyses additionally adjusted for age and days in milk. Holstein cows had greater body weight, dry matter intake, milk yield, fat- and protein-corrected milk, and daily CH₄ production than Jersey cows. Jersey cows had higher milk fat and protein concentrations, whereas daily milk fat yield did not differ significantly between breeds. No significant breed differences were detected in CH₄ production per unit of dry matter intake, milk yield, or fat- and protein-corrected milk. In the unadjusted analysis, Jersey cows had lower CH₄ production per unit of milk fat yield, but the breed difference was inconclusive after adjustment for age and days in milk. Methane production per unit of milk protein or lactose yield did not differ significantly between breeds. These findings indicate that breed comparisons of enteric CH₄ depend on the basis used to express emissions, while the component-specific ratios should be interpreted as exploratory animal-level measures. Full article

(This article belongs to the Special Issue Advances in Measuring and Mitigating Methane Emissions from Ruminants)

29 pages, 2156 KB

Open AccessReview

A Narrative Review of Early Pregnancy Diagnosis Technologies for Livestock: From Conventional to Intelligent Systems

by Yang Shen, Yujie Zhang, Junyi Meng, Yutong Han, Jitong Xu, Hongying Wang and Liangju Wang

Animals 2026, 16(18), 2897; https://doi.org/10.3390/ani16182897 - 15 Sep 2026

Abstract

Accurate and efficient early pregnancy diagnosis (EPD) in livestock is crucial for optimizing breeding management and enhancing productivity in modern animal husbandry. Over the past century, EPD technology has evolved from empirical methods to sophisticated techniques, encompassing biochemical marker detection, ultrasonic imaging, and [...] Read more.

Accurate and efficient early pregnancy diagnosis (EPD) in livestock is crucial for optimizing breeding management and enhancing productivity in modern animal husbandry. Over the past century, EPD technology has evolved from empirical methods to sophisticated techniques, encompassing biochemical marker detection, ultrasonic imaging, and further extending to emerging non-invasive approaches such as infrared thermography (IRT) and spectroscopic analysis. These advancements have not only improved diagnostic accuracy but also broadened the research scope to include small livestock and multiple species. This review critically examines the historical evolution, current methodologies, and applications of EPD technology, with a focus on analyzing the advantages and limitations of both traditional and emerging techniques. Additionally, it explores the potential of multimodal fusion strategies and artificial intelligence (AI) in EPD. At present, machine vision, wearable monitoring, and several AI applications remain prospective approaches rather than validated tools for routine EPD. The conclusion highlights that, despite significant progress, current technologies still face limitations in achieving in situ, non-contact, and high-throughput detection. Looking ahead, the integration of cutting-edge technologies, such as AI, small wearable sensors, and physiological time-series data analysis, holds promise for overcoming these bottlenecks, enabling more intelligent and efficient pregnancy diagnosis, and providing scientific support for modern animal husbandry. Full article

(This article belongs to the Section Animal System and Management)

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14 pages, 5827 KB

Open AccessArticle

Evaluation of Two Commercial DNA-Based Breed Identification Tests for Pit Bull-Type Dogs in Taiwan: A Pilot Study

by Yung-Yun Yang, Ann Nee Lee, Tzu-Chieh Chen and Wei-Hsiang Huang

Animals 2026, 16(18), 2896; https://doi.org/10.3390/ani16182896 - 15 Sep 2026

Abstract

The “Declaration of Specified Breed Names of Animals Banned from Being Owned, Imported, or Exported” was promulgated in Taiwan in response to aggression involving pit bull-type dogs. To support enforcement, we developed standards for visual breed identification based on international breed standards and [...] Read more.

The “Declaration of Specified Breed Names of Animals Banned from Being Owned, Imported, or Exported” was promulgated in Taiwan in response to aggression involving pit bull-type dogs. To support enforcement, we developed standards for visual breed identification based on international breed standards and UK guidance. Given the subjectivity of visual identification, samples from 21 healthy dogs, including 12 pit bull-type dogs, were submitted to Embark^® and Orivet Genetic Pet Care for SNP-based breed identification. Across pit bull-type dogs, American Pit Bull Terrier signatures reported by Embark^® were reported as American Staffordshire Terrier by Orivet, while Formosan Mountain Dog signatures were reported as mixed breeds. Embark^® covered all four pit bull-type breeds and the Formosan Mountain Dog and produced fewer failed tests, providing broader coverage of breeds relevant to Taiwan. However, substantial discordance was observed between the two services in the assignment of the American Pit Bull Terrier and American Staffordshire Terrier, the two breeds specified in the Taiwanese declaration. This findings suggest that commercial DNA breed identification requires further validation before it can support regulatory enforcement. Full article

(This article belongs to the Section Public Policy, Politics and Law)

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24 pages, 10705 KB

Open AccessArticle

Integrated Biological Responses Define the Thermoneutral Zone of Murrah Buffalo Calves in a Subtropical Climate

by Reetu Kumari, Priyambada Kumari, Brijesh Yadav, Shanker Kumar Singh, Vansh Sharma, Manish Tiwari and Arun Kumar Madan

Animals 2026, 16(18), 2895; https://doi.org/10.3390/ani16182895 - 14 Sep 2026

Abstract

The study was conducted to establish the thermoneutral zone (TNZ) of Murrah buffalo calves under precisely regulated environmental conditions. Six healthy calves (8–10 months) were exposed to progressively decreasing (24–15 °C; Temperature-Humidity Index (THI): 72.45–58.98) and increasing (25–40 °C; THI: 73.49–90.32) ambient temperatures [...] Read more.

The study was conducted to establish the thermoneutral zone (TNZ) of Murrah buffalo calves under precisely regulated environmental conditions. Six healthy calves (8–10 months) were exposed to progressively decreasing (24–15 °C; Temperature-Humidity Index (THI): 72.45–58.98) and increasing (25–40 °C; THI: 73.49–90.32) ambient temperatures with 3 °C intervals in a psychrometric chamber. Each exposure was maintained for 10 consecutive days under cyclic conditions (12 h exposure and 12 h thermoneutral recovery). Physiological responses and body surface temperature (BST) were recorded, and blood samples were collected on day 10 at 1500 h. Segmented regression analysis was performed using the SegReg (Oosterbaan, 2017) software program to identify lower and upper critical temperatures (LCT, UCT) and corresponding THI thresholds. LCTs ranged from 18.06 to 19.14 °C across different responsive parameters, with breakpoints (BPs) at 18.60 °C (SE_BP = 0.634) for pulse rate (PR), 18.15 °C (SE_BP = 0.945) for haemoglobin, 18.15 °C (SE_BP = 0.189) for lymphocytes, 18.24 °C (SE_BP = 0.404) for granulocytes, 19.14 °C (SE_BP = 1.09) for reactive oxygen species (ROS), 18.15 °C (SE_BP = 23.9) for superoxide dismutase (SOD), and 18.06 °C (SE_BP = 4.2) for cortisol; no distinct LCT was detected for respiratory rate (RR), rectal temperature (RT), total leukocyte count (TLC), or heat shock protein (HSP) 70/HSP90 expression. BST at different locations exhibited an LCT between ~20 and 21 °C. UCTs for PR, RR and RT were observed at 28.9 °C (SE_BP = 0.892), 28.6 °C (SE_BP = 0.843) and 29.2 °C (SE_BP = 1.16), respectively. Neither BST nor erythrocytic parameters exhibited a distinct UCT; however, BST increased progressively with increasing exposure temperature. TLC, lymphocyte and granulocyte % showed UCTs of 28.15 °C (SE_BP = 0.753), 28.15 °C (SE_BP = 0.497) and 32.65 °C (SE_BP = 14.20), respectively. The UCT for cortisol was observed at 28.15 °C (SE_BP = 5.52), whereas ROS and SOD showed UCTs of 28.75 °C (SE_BP = 4.67) and 34.15 °C (SE_BP = 6.55), respectively. HSP70 and HSP90 exhibited breakpoints at 35.35 °C (SE_BP = 1.32) and 34.30 °C (SE_BP = 2.11), respectively. Integration of key physiological and systemic responses (excluding BST) indicated a TNZ ranging from 19.14 °C to 28.15 °C. These findings provide a multi-level characterization of thermophysiological responses in buffalo calves and support the use of integrated biological indicators for defining thermoneutral conditions under controlled environments. The estimated TNZ of 19.14–28.15 °C provides a practical reference for managing Murrah buffalo calves. Producers should monitor housing temperature and consider ventilation, shade and air movement when temperatures approach or exceed ~28 °C while providing protection from cold when temperatures fall below ~19 °C. This may help reduce thermal strain and maintain normal physiological function. Full article

(This article belongs to the Section Animal Physiology)

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12 pages, 495 KB

Open AccessArticle

Distilled Water Dilution Increases Measured Fecal pH in Fresh Canine Feces

by Pedro Henrique Marchi, Carolina Dantas Micheletti, Jennifer Machado de Souza, Gabriela Luiza Fagundes Finardi, Eduarda Lorena Fernandes, Nicholas Sartini Santiago, Luana Dias dos Santos, Matheus Henrique dos Santos, Júlio Cesar de Carvalho Balieiro and Thiago Henrique Annibale Vendramini

Animals 2026, 16(18), 2894; https://doi.org/10.3390/ani16182894 - 14 Sep 2026

Abstract

Fecal pH is used in canine nutrition studies as an indirect indicator of intestinal fermentation, but sample preparation may affect the measured value. This study evaluated the effect of different feces-to-water dilution ratios on pH determination in canine feces. Thirty-seven samples from 20 [...] Read more.

Fecal pH is used in canine nutrition studies as an indirect indicator of intestinal fermentation, but sample preparation may affect the measured value. This study evaluated the effect of different feces-to-water dilution ratios on pH determination in canine feces. Thirty-seven samples from 20 healthy dogs were evaluated using five preparation protocols: direct measurement without dilution and dilution with distilled water at 1:1, 1:2, 1:5, and 1:10 feces-to-water ratios, on a weight-to-volume basis. Sample preparation affected fecal pH (p < 0.0001). The least-squares mean pH was 6.37 for direct measurement, 6.54 for 1:1, 6.71 for 1:2, 6.88 for 1:5, and 7.04 for 1:10. Fecal pH increased progressively with increasing water addition, with significant differences among all preparation protocols. Regression analysis confirmed that water addition increased measured fecal pH, and the response was better described by a quadratic model than by a linear model. These findings show that the amount of water added is a relevant methodological factor in fecal pH determination. Among the protocols evaluated, the 1:1 feces-to-water ratio appears to be the most suitable standardized method for routine analysis because it used the lowest amount of water while providing adequate conditions for homogenization and electrode contact. Full article

(This article belongs to the Special Issue Small Animal Nutrition and Health)

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23 pages, 2517 KB

Open AccessArticle

Integrated Bulk and Single-Nucleus Transcriptomics Characterizes the Developmental and Regional Specialization of Chicken Abdominal Fat-Associated Tissue

by Siyu Zhang, Qihong Zhang, Jiali Wang and Zhenhui Li

Animals 2026, 16(18), 2893; https://doi.org/10.3390/ani16182893 - 14 Sep 2026

Abstract

Excessive abdominal fat deposition reduces feed efficiency and edible carcass yield in chickens, yet its developmental and regional specialization remains poorly understood. We generated 93 bulk RNA sequencing (RNA-seq) libraries from 15 fat-associated anatomical regions at embryonic day 16 (E16) and 16 regions [...] Read more.

Excessive abdominal fat deposition reduces feed efficiency and edible carcass yield in chickens, yet its developmental and regional specialization remains poorly understood. We generated 93 bulk RNA sequencing (RNA-seq) libraries from 15 fat-associated anatomical regions at embryonic day 16 (E16) and 16 regions at 2 weeks post-hatch (2w) in six Xinghua chickens. Exploratory single-nucleus RNA sequencing was additionally performed on abdominal region (AR) tissue from one individual at each stage. Repeated-measures-aware analysis identified developmental stage as the major source of transcriptomic variation, with 4556 genes expressed at higher levels at E16 and 4528 at 2w. E16-associated programs were enriched for proliferation, morphogenesis, and structural development, whereas postnatal programs were dominated by immune and stimulus-response processes. Stage-specific weighted gene co-expression network analysis (WGCNA) identified substantial regional heterogeneity. AR showed stage-dependent signatures, including higher developmental and morphogenetic programs and lower lipid-metabolic programs at E16, and higher cell-cycle-related and lower developmental and fatty-acid-metabolic programs at 2w. Single-nucleus analysis identified adipose-lineage and multiple non-adipocyte populations, and cross-omics mapping showed that AR-associated signatures were distributed across multiple cellular compartments. These results suggest stage-dependent, multicellular transcriptional specialization of chicken AR tissue. Full article

(This article belongs to the Section Animal Genetics and Genomics)

21 pages, 3356 KB

Open AccessArticle

Identifying Key Factors Associated with the Spatiotemporal Dynamics of Fish Assemblages in Subtropical River-Type Reservoir Ecosystems: Implications for Fishery Management and Conservation

by Chengjie Yin, Rongzhen Zhang, Zewen Feng, Xiaoning Shi, Renhui Li, Miao Xiang, He Zhang and Longgen Guo

Animals 2026, 16(18), 2892; https://doi.org/10.3390/ani16182892 - 14 Sep 2026

Abstract

Characterizing the spatiotemporal patterns of fish communities in subtropical river-type reservoirs provides useful baseline information for fisheries stewardship and aquatic ecosystem monitoring. This work investigated fish assemblages across 18 sampling sites within Shanxi Reservoir, Zhejiang Province, using an integrative approach combining gillnet surveys, [...] Read more.

Characterizing the spatiotemporal patterns of fish communities in subtropical river-type reservoirs provides useful baseline information for fisheries stewardship and aquatic ecosystem monitoring. This work investigated fish assemblages across 18 sampling sites within Shanxi Reservoir, Zhejiang Province, using an integrative approach combining gillnet surveys, hydroacoustic detection, and habitat parameter monitoring. Sampling was conducted during two periods: October 2022 (autumn, post-flood, fattening phase) and April 2023 (spring, normal water level, spawning phase). A total of 22 species were documented in spring and 21 in autumn, with Cyprinidae species predominating. Dominant species shifted seasonally, led by Hypophthalmichthys nobilis, Culter dabryi and Hemiculter leucisculus. Planktivores and carnivores were more abundant in spring, while omnivores increased in autumn. Spatially, fish populations were mainly concentrated in the shallow riverine zone (SRZ), predominantly comprising small- to medium-sized individuals. Mantel tests and Pearson correlation analyses identified associations between fish assemblage structure in the deep lake zone (DLZ) and environmental variables including dissolved oxygen, conductivity, chlorophyll a, and phytoplankton density and biomass. This study provides preliminary baseline data on fish community dynamics in Shanxi Reservoir and offers tentative observations that may help inform future zoned monitoring and ecological restoration planning. Full article

(This article belongs to the Special Issue Fish and Fisheries Under Ecosystem Changes)

18 pages, 5770 KB

Open AccessArticle

Milking-Order Stability and Stabilization After Herd Entry Under Heat Stress: An Observational Study in a Large Dairy Herd

by Yifeng Song, Zhalaga, Muqier, Xiaoping An, Na Liu and Jingwei Qi

Animals 2026, 16(18), 2891; https://doi.org/10.3390/ani16182891 - 14 Sep 2026

Abstract

Heat stress is known to affect dairy cow behavior, but its effects on milking-order patterns and stabilization after herd entry in large commercial herds are not well described. We analyzed 70,491 parlor records from 484 Holstein cows milked three times daily in a [...] Read more.

Heat stress is known to affect dairy cow behavior, but its effects on milking-order patterns and stabilization after herd entry in large commercial herds are not well described. We analyzed 70,491 parlor records from 484 Holstein cows milked three times daily in a free-stall herd to examine how thermal conditions shape milking-order dynamics. Daily intraday milking-order displacement was quantified based on changes in queue position between successive milkings. Cow-specific regressions of milking order on ambient temperature were used to identify clusters of thermal response, and time to reach a stable queue position was evaluated for newly introduced cows according to thermal conditions at herd entry. Results show a modest increase in intraday physical displacement from thermally comfortable to severely hot days, whereas relative displacement did not differ significantly across the four thermal categories. Cows showed consistent individual differences in how their milking position shifted with temperature that were only weakly associated with parity, days in milk, milk yield, or panting score. In an exploratory severe versus non-severe comparison, cows entering the herd under severe thermal conditions had a lower adjusted hazard of reaching the predefined stabilization criterion (HR = 0.62, 95% CI: 0.43–0.90). Collectively, these findings indicate that milking-order patterns extracted from routine parlor records are associated with behavioral responses to thermal conditions. Their management implications, including decisions related to herd entry and cooling, require further experimental evaluation. Full article

(This article belongs to the Section Cattle)

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20 pages, 266 KB

Open AccessArticle

“Learning to Live Without Him in My Home When a Piece of My Heart Went with Him”: End-of-Life Decision-Making Experiences of Companion Animal Caregivers

by Lori R. Kogan, Jennifer Currin-McCulloch, Sohaila Jafarian, Emma Klein and Kathleen Cooney

Animals 2026, 16(18), 2890; https://doi.org/10.3390/ani16182890 - 14 Sep 2026

Abstract

As companion animals approach end of life, their caregivers often face complex decisions that require balancing their pet’s quality of life, relief from suffering, potential medical interventions, financial and resource barriers, and the difficult question of whether and when to pursue euthanasia. The [...] Read more.

As companion animals approach end of life, their caregivers often face complex decisions that require balancing their pet’s quality of life, relief from suffering, potential medical interventions, financial and resource barriers, and the difficult question of whether and when to pursue euthanasia. The purpose of this qualitative study was to explore how companion animal caregivers experience end-of-life decision-making for their pet and the care they received from their veterinary team. The study examined the experiences of 143 pet caregivers who responded to an online survey between September and December 2025. Data collection included caregiver’s responses to four open-ended prompts inquiring about their experiences with their pet’s end-of-life. Questions explored what they regarded as the most challenging and meaningful aspects, regrets they held about their pet’s death, and advice for veterinarians about how best to support pet owners in their pet’s final moments. Data were analyzed using Braun and Clarke’s reflexive thematic analysis. Results highlighted caregivers’ distress surrounding decisions about whether and when they should pursue euthanasia. Caregivers described balancing multiple considerations in their decision-making, including their pet’s overall welfare, the needs and perspectives of family members, resource constraints, access to specialized veterinary care, and their own caregiving fatigue. Their responses demonstrated grief, regret, guilt, and meaning making. Veterinarians’ compassion, transparency, and forward-thinking enabled caregivers to plan for rituals and meaningful death experiences for their pets and their families. Suggestions for veterinary care emphasized the importance of relationship-centered care in addition to technical expertise. Veterinary end-of-life care should prioritize empathy, early and transparent communication, and opportunities for cherished time during the pet’s final moments. Full article

(This article belongs to the Special Issue The Human–Animal Bond in Clinical Practice)

17 pages, 8204 KB

Open AccessArticle

Impacts of Sprouted Barley on Performance and Carcass Ultrasound Measurements in Angus-Influenced Steers During Backgrounding

by Zachary C. Crump, Anthony F. Alberto, Bailee L. Brown, Bryce W. Roholt, Lillian L. Okamoto, Ryan A. Spurling, Robert E. Ward, Brady M. Blackett, Robert L. Harding, Sanjeewa D. Ranathunga and Kara J. Thornton

Animals 2026, 16(18), 2889; https://doi.org/10.3390/ani16182889 - 14 Sep 2026

Abstract

The objective of this research was to determine the impacts of including sprouted barley (SB) in the diets of backgrounding beef steers. Angus-influenced steers (328 ± 6.59 kg) were assigned to one of four diets (n = 15/diet): a control (CON) diet, or [...] Read more.

The objective of this research was to determine the impacts of including sprouted barley (SB) in the diets of backgrounding beef steers. Angus-influenced steers (328 ± 6.59 kg) were assigned to one of four diets (n = 15/diet): a control (CON) diet, or a diet containing 5%, 10%, or 20% SB (DM basis)—replacing corn silage and some rolled barley. Individual intake and feeding behavior were monitored using Vytelle bunks. Body weight (BW) and ruminal fluid samples were collected on d 0, 30, and 60, while dry matter intake (DMI), feeding behavior, average daily gain (ADG), and gain-to-feed ratio (G:F) were monitored for 60 d. In addition, total amount of nutrients consumed (crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), net energy for maintenance (NEm), and net energy for gain (NEg)) were also assessed. Calf BW, ADG, and G:F were not impacted by diet (p ≤ 0.43). DMI and consumption of CP, NDF, ADF, NEm and NEg were all impacted by diet × day (p ≤ 0.01). The acetate:propionate, percentage of propionate and concentration of butyrate were each impacted by diet × day (p ≤ 0.007). Collectively, these findings suggest including SB in the diet modifies nutrient intake and ruminal fermentation in backgrounding steers. Full article

(This article belongs to the Section Animal Nutrition)

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19 pages, 1570 KB

Open AccessArticle

Analysis of Zona-Associated Sperm as a Potential Source of Exogenous Y-DNA Interference in Sex-Typing of In Vitro-Fertilized Ovine Embryos

by Qiangqiang Ma, Ying Chen, Hong Dong, Liqin Wang, Xuefeng Fu, Gulimire Abudureyimu, Jiaxin Zhang, Wei Zhang, Jinbo Tian, Yangsheng Wu and Jiapeng Lin

Animals 2026, 16(18), 2888; https://doi.org/10.3390/ani16182888 - 14 Sep 2026

Abstract

This study aimed to explore whether zona-associated residual sperm may contribute to exogenous Y-DNAY-DNA contamination and cause sex bias in PCR-based sex classification of ovine early-stage embryos, based on the amplification of the AMEL gene. Primers targeting the conserved regions of the ovine [...] Read more.

This study aimed to explore whether zona-associated residual sperm may contribute to exogenous Y-DNAY-DNA contamination and cause sex bias in PCR-based sex classification of ovine early-stage embryos, based on the amplification of the AMEL gene. Primers targeting the conserved regions of the ovine AMELX and AMELY genes were designed, and a nested PCR system using AMEL and YWHAZ genes was established. Group A comprised 17 female and 33 male embryos, corresponding to a female-to-male ratio of 0.52:1, whereas Group B comprised 23 female and 27 male embryos, corresponding to a female-to-male ratio of 0.85:1. DAPI staining revealed sperm-morphology structures or structures containing sperm-derived DNA adjacent to the zona pellucida of ovine early-stage embryos. Sequence alignment of PCR amplicons showed consistency with the AMELX and AMELY reference genes. In conclusion, these results support our research hypothesis that zona-associated residual sperm may serve as a potential source of Y-DNAY-DNA contamination and interfere with PCR-based embryonic sex determination. Full article

(This article belongs to the Topic Advances in Animal Reproductive Physiology and Technologies)

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15 pages, 1094 KB

Open AccessArticle

Fasting Serum Bile Acid Profiles in Dogs with Food-Responsive and Immunosuppressant-Responsive Chronic Inflammatory Enteropathy: An Exploratory LC–MS/MS Study

by Gergely Kiss, Borbála Mózes, Ágnes Sterczer, Katalin Lányi, Krisztián Németh and Roland Psáder

Animals 2026, 16(18), 2887; https://doi.org/10.3390/ani16182887 - 14 Sep 2026

Abstract

Bile acid (BA) dysmetabolism is implicated in canine chronic inflammatory enteropathy (CIE), but fasting serum profiles across treatment-response groups remain poorly characterized. This exploratory observational study analyzed pretreatment serum from dogs subsequently classified as food-responsive enteropathy (FRE; n = 8) or immunosuppressant-responsive enteropathy [...] Read more.

Bile acid (BA) dysmetabolism is implicated in canine chronic inflammatory enteropathy (CIE), but fasting serum profiles across treatment-response groups remain poorly characterized. This exploratory observational study analyzed pretreatment serum from dogs subsequently classified as food-responsive enteropathy (FRE; n = 8) or immunosuppressant-responsive enteropathy (IRE; n = 5), with four unmatched purpose-bred Beagle controls. Targeted LC–MS/MS, performed using a published canine serum method, provided results for 15 BA species. All samples were analyzed in one batch by an analyst blinded to subsequent group assignment. Eight species quantifiable in every dog and ten derived variables were compared using Kruskal–Wallis tests with Benjamini–Hochberg correction across 18 omnibus tests. Cholic acid and unconjugated primary BA concentrations were higher in IRE than in controls and FRE. The total primary-to-secondary BA ratio showed a monotonic group ordering (control < FRE < IRE; H = 13.76, p = 0.0010, q = 0.0092; epsilon-squared = 0.840), with all Bonferroni-corrected pairwise comparisons significant. A post hoc four-scenario sensitivity analysis comprising the submitted zero-substitution analysis and three LOD/LOQ-based alternatives reproduced the same 18-test FDR-significant set and the same ratio inference. This analysis tests robustness to censored-value handling but is not independent validation. The findings suggest group-associated serum BA patterns, but the small sample, clinical heterogeneity, and unmatched controls preclude diagnostic or treatment-predictive interpretation. Full article

(This article belongs to the Section Companion Animals)

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15 pages, 979 KB

Open AccessArticle

Effects of SIRT7 Inhibition on Milk Fat Synthesis and Fatty Acid Composition in Dairy Goat Mammary Epithelial Cells

by Jun Li, Zhongying Li, Menghan Han, Mengfan Zhao, Zhuoying Li, Huibin Shi, Haoyuan Han, Kun Liu, Xing Liu and Kai Quan

Animals 2026, 16(18), 2886; https://doi.org/10.3390/ani16182886 - 14 Sep 2026

Abstract

Milk fat, a key nutritional component of milk, not only supplies energy and immune protection for offspring but also serves as a critical quality indicator for goat milk. Sirt7, one of the least studied members of the Sirtuins family, has been shown to [...] Read more.

Milk fat, a key nutritional component of milk, not only supplies energy and immune protection for offspring but also serves as a critical quality indicator for goat milk. Sirt7, one of the least studied members of the Sirtuins family, has been shown to play a regulatory role in lipid metabolism. This study investigated the regulatory role of Sirt7 in milk fat synthesis and fatty acid composition in dairy goat mammary epithelial cells (GMECs) using a Sirt7 inhibitor (ID: 97491). GMECs were treated with a Sirt7 inhibitor (ID: 97491), and cell viability was assessed via the CCK-8 assay, which confirmed that 15 μmol/L inhibitor had no cytotoxic effects. Functional analyses using RT-qPCR, Western blot, and GC-MS revealed that Sirt7 inhibition upregulated the expression of SREBP1, CD36, ACC, SCD1, and ELOVL6 while downregulating FOXO1 and FABP1. At the protein level, the inhibitor reduced Sirt7, FOXO1. Additionally, Sirt7 inhibition increased intracellular lipid droplet and triglyceride content. GC-MS analysis showed that Sirt7 inhibition decreased C11:0; increased C16:0, C15:0, C20:0, C24:0, C16:1, and C18:1n9c; and decreased C20:5n3 (EPA). Collectively, these findings suggest that pharmacological inhibition of Sirt7 is associated with upregulation of SREBP1, disruption of lipid homeostasis, triglyceride accumulation, and altered fatty acid composition in GMECs. These results provide preliminary evidence for a potential regulatory role of Sirt7 in milk fat synthesis and lay a foundation for future mechanistic studies. Full article

(This article belongs to the Section Animal Genetics and Genomics)

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16 pages, 598 KB

Open AccessArticle

University-Based Animal Welfare Organizations in Ecuador: A National One Health Perspective

by Ángel Sebastián Rodríguez-Pazmiño, Sarita Doménica Bautista-Arcentales, Miguel Angel Garcia-Bereguiain and Solon Alberto Orlando

Animals 2026, 16(18), 2885; https://doi.org/10.3390/ani16182885 - 14 Sep 2026

Abstract

University-based animal welfare organizations are grassroots actors involved in managing free-roaming dog and cat populations, yet their role remains poorly characterized, particularly in low- and middle-income countries. This study aimed to identify and describe animal welfare organizations operating within universities in Ecuador and [...] Read more.

University-based animal welfare organizations are grassroots actors involved in managing free-roaming dog and cat populations, yet their role remains poorly characterized, particularly in low- and middle-income countries. This study aimed to identify and describe animal welfare organizations operating within universities in Ecuador and to analyze their potential contributions within a One Health framework. A cross-sectional survey of 63 universities was conducted using publicly available information from institutional websites and social media platforms, complemented by semi-structured interviews with representatives of identified organizations. Organizations with evidence of activity through 2026 and a focus on dogs or cats were included. A total of 14 animal welfare organizations from 12 universities were identified, representing 19.0% of the universities analyzed. Most organizations were located in public universities (75.0%), and 28.6% were associated with veterinary-related programs. Thematic analysis of interview data (12 out of 14 organizations interviewed) revealed four key dimensions of organizational functioning: capacity and governance, animal care and population management, public health and community engagement, and institutional integration. Reported activities included animal rescue, sterilization campaigns, adoption initiatives, and community outreach, aligning with internationally recommended strategies for humane population management. Several organizations have extended their reach beyond campus boundaries through free veterinary intervention campaigns in rural communities, partnerships with municipal authorities, and collaboration with civil society networks, reflecting a broader public health role consistent with One Health principles. Despite their potential, these organizations face structural challenges, including high volunteer turnover, the absence of direct institutional funding, and limited formal integration into governmental frameworks. Institutional support, where present, is predominantly material rather than financial, placing the burden of sustainability on self-management mechanisms. Nonetheless, their embeddedness within academic environments positions them as strategic platforms for interdisciplinary collaboration, experiential training, and community engagement. This study highlights the need to recognize and integrate university-based animal welfare organizations into broader animal population management and public health strategies. Strengthening partnerships between academia, government, and civil society could enhance both animal welfare outcomes and zoonotic disease surveillance efforts in Ecuador and other low- and middle-income countries. Full article

(This article belongs to the Special Issue One Health and Public Health Implications of Sterilization-Based Management in Free-Roaming Dogs and Cats)

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31 pages, 40449 KB

Open AccessArticle

A Multi-Object Tracking Method for Dairy Cows in Intensive Farming Scenarios

by Zhihua Diao, Zhichao Huang, Jiangbo Li, Jinpeng Cheng, Suna Zhao and Baohua Zhang

Animals 2026, 16(18), 2884; https://doi.org/10.3390/ani16182884 - 13 Sep 2026

Abstract

To better analyze the health and welfare of individual dairy cows, this study proposes BR-Tracker, a multi-object tracking method designed for dense monitoring environments to address missed detections, tracking failures, and frequent identity switches. In the object detection stage, a Receptive Field Attention [...] Read more.

To better analyze the health and welfare of individual dairy cows, this study proposes BR-Tracker, a multi-object tracking method designed for dense monitoring environments to address missed detections, tracking failures, and frequent identity switches. In the object detection stage, a Receptive Field Attention Downsampling (RFADown) module is introduced into the neck network of YOLOv10s to effectively process the locally visible regions of occluded cows by dynamically adjusting the receptive field. An improved Partial Bi-Level Routing Attention (PBRA) module is incorporated into the backbone network to simultaneously extract global and local features, while a Spatial Pyramid Pooling with Efficient Layer Aggregation Network (SPPELAN) module is adopted to enhance multi-scale feature aggregation. In the object tracking stage, an MPDIoU-based matching algorithm is designed to improve matching accuracy and the reliability of trajectory association. The dataset contains 15,420 images for object detection and 130 independent videos for multi-object tracking, of which 40 videos are selected for the final tracking evaluation. Experimental results show that BR-YOLOv10s achieves a precision, recall, and mean average precision (mAP) of 95.7%, 90.3%, and 95.2%, respectively. Compared with the YOLOv10s-ByteTrack baseline, the proposed method improves HOTA, MOTA, MOTP, and IDF1 by 4.4, 5.7, 3.0, and 6.1 percentage points, respectively, while reducing identity switches by 23.19%. In addition, the tracker achieves a processing speed of 43.2 FPS. These results demonstrate that the proposed method can effectively perform real-time detection and tracking of densely distributed dairy cows in complex intensive farming environments. Full article

(This article belongs to the Collection Monitoring of Cows: Management and Sustainability)

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