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Search Results (3,623)

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14 pages, 2574 KiB  
Article
Protective Effects of Chitosan Oligosaccharide Against Lipopolysaccharide-Induced Inflammatory Response and Oxidative Stress in Bovine Mammary Epithelial Cells
by Ziwei Lin, Yanlong Zhou, Ruiwen Chen, Qiuyan Tao, Qiwen Lu, Qianchao Xu, Haibin Yu, Ping Jiang and Zhihui Zhao
Mar. Drugs 2025, 23(1), 31; https://doi.org/10.3390/md23010031 - 9 Jan 2025
Viewed by 197
Abstract
Chitosan oligosaccharide (COS) is receiving increasing attention as a feed additive in animal production. COS has a variety of biological functions, including anti-inflammatory and antioxidant activities. Mastitis is a major disease in dairy cows that has a significant impact on animal welfare and [...] Read more.
Chitosan oligosaccharide (COS) is receiving increasing attention as a feed additive in animal production. COS has a variety of biological functions, including anti-inflammatory and antioxidant activities. Mastitis is a major disease in dairy cows that has a significant impact on animal welfare and production. Hence, this research aimed to investigate the mechanism of COS on the lipopolysaccharide (LPS)-stimulated inflammatory response and oxidative stress in bovine mammary epithelial cells (BMECs). In this study, the results demonstrated that COS protected BMECs from the inflammatory response induced by LPS by restraining the excessive production of toll-like receptor 4 (TLR4), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). COS treatment also suppressed excessive reactive oxygen species (ROS) production and restored antioxidant enzyme activity under LPS-induced oxidative stress conditions. Furthermore, the results also demonstrated that COS promote nuclear factor erythroid 2-related factor 2 (Nrf2) expression and inhibit TLR4 levels and p65 and IκBα phosphorylation in BMECs exposed to LPS. In summary, the results demonstrate that the protective mechanism of COS on the LPS-induced inflammatory response and oxidative stress depend on the TLR4/nuclear factor-κB (NF-κB) and Nrf2 signaling pathways, indicating that COS could serve as natural protective agents for alleviating BMECs in mastitis. Full article
(This article belongs to the Special Issue Marine Natural Products and Signaling Pathways, 2nd Edition)
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Figure 1
<p>Effect of COS and LPS on BMEC viability. (<b>A</b>) Viability of BMECs treated with 150 μg/mL COS for 36 h. (<b>B</b>) Viability of BMECs pretreated with 150 μg/mL COS for 12 h and co-treated with 10 μg/mL LPS for an additional 24 h. Data are presented as the mean ± standard deviation (SD) (<span class="html-italic">n</span> = 3). ** <span class="html-italic">p</span> &lt; 0.01 indicates a statistically significant difference.</p>
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<p>COS alleviated the LPS-induced inflammatory response in BMECs. The mRNA levels of (<b>A</b>) <span class="html-italic">TLR4</span>, (<b>B</b>) <span class="html-italic">TNF-α</span>, (<b>C</b>) <span class="html-italic">IL-1β</span>, and (<b>D</b>) <span class="html-italic">IL-6</span> were assessed using real-time quantitative PCR (RT-qPCR). The protein expression of (<b>E</b>) TLR4, (<b>F</b>) TNF-α, (<b>G</b>) IL-1β, and (<b>H</b>) IL-6 was tested using ELISAs. Data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Groups with different superscript letters were considered statistically different (<span class="html-italic">p</span> &lt; 0.05)).</p>
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<p>COS mitigated LPS-induced oxidative stress in BMECs. (<b>A</b>) Effects of COS on ROS fluorescence intensities in LPS-induced BMECs. Fluorescence microscopic images showing ROS production. (<b>B</b>) The fluorescence intensities were analyzed using Image J. (<b>C</b>,<b>D</b>) SOD and CAT activities. (<b>E</b>) <span class="html-italic">Nrf2</span> mRNA expression as assessed with RT-qPCR. Scale bar = 100 μm. Data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Groups with different superscript letters are statistically different (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>COS alleviated oxidative stress and inflammatory response induced by LPS by regulating Nrf2 and the TLR4/NF-κB signaling pathway. (<b>A</b>) Western blot (WB) images of Nrf2. (<b>B</b>) Quantification analysis of Nrf2 expression. (<b>C</b>) WB images of TLR4, p-IκBα, IκBα, p-p65, and p65. The quantification analysis of (<b>D</b>) TLR4, (<b>E</b>) p-IκBα/IκBα, and (<b>F</b>) p-p65/p65 expression. Data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Groups with different superscript letters are considered statistically different (<span class="html-italic">p</span> &lt; 0.05).</p>
Full article ">Figure 4 Cont.
<p>COS alleviated oxidative stress and inflammatory response induced by LPS by regulating Nrf2 and the TLR4/NF-κB signaling pathway. (<b>A</b>) Western blot (WB) images of Nrf2. (<b>B</b>) Quantification analysis of Nrf2 expression. (<b>C</b>) WB images of TLR4, p-IκBα, IκBα, p-p65, and p65. The quantification analysis of (<b>D</b>) TLR4, (<b>E</b>) p-IκBα/IκBα, and (<b>F</b>) p-p65/p65 expression. Data are presented as the mean ± SD (<span class="html-italic">n</span> = 3). Groups with different superscript letters are considered statistically different (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Protective mechanisms of COS in LPS-induced BMECs (image generated using Figdraw).</p>
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9 pages, 872 KiB  
Article
Rearing in a Physically Enriched Environment Affects Shoaling and Stress Responses of Zebrafish (Danio rerio) Exposed to Novel Conditions
by Valentina Gazzano, Martina Di Filippo, Rosario Licitra, Asahi Ogi, Baldassare Fronte, Maria Claudia Curadi and Angelo Gazzano
Vet. Sci. 2025, 12(1), 38; https://doi.org/10.3390/vetsci12010038 - 9 Jan 2025
Viewed by 257
Abstract
The impact of enrichment on stress reduction in zebrafish (Danio rerio) exposed to a novel environment was assessed. Four control shoals (CTRL) and five treated shoals (TRT), each with eight fish, were observed; in TRT tanks, a PVC pipe was included [...] Read more.
The impact of enrichment on stress reduction in zebrafish (Danio rerio) exposed to a novel environment was assessed. Four control shoals (CTRL) and five treated shoals (TRT), each with eight fish, were observed; in TRT tanks, a PVC pipe was included (three-way tube, 11.7 × 4 cm) as enrichment for 90 days. Subsequently, fish were moved to a new tank for a shoaling test, and behavior was evaluated over periods of 0′–5′ and 5′–10′. Cortisol dissolved in water was measured before and after the test. No differences were found between the two groups in distance moved, swimming speed, or shoal acceleration. Both groups reduced interindividual distance in the second phase of the test (CTRL: t = 8.977, p ≤ 0.0001; TRT: t = 8.247, p ≤ 0.0001), though TRT fish maintained greater spacing (t = 2.292, p ≤ 0.05). TRT fish spent more time without contact during both phases (first: t = 2.645, p ≤ 0.05; second: t = 3.134, p ≤ 0.01), while CTRL fish reduced this time in the second phase (t = 2.991, p ≤ 0.05). Cortisol rose significantly in CTRL after the test (t = 2.452, p ≤ 0.05) but not in TRT fish. These results suggest that environmental enrichment mitigates stress, as seen by reduced cohesiveness and cortisol in TRT fish. Full article
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<p>Enrichment employed: 3-way orange PVC pipe modified (Tecnomat<sup>®</sup>).</p>
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<p>Shoaling behavior (<b>A</b>–<b>E</b>) and water cortisol concentrations (<b>F</b>) in the control (CTRL; <span class="html-italic">n</span> = 4 tanks of 8 fish) and treated (TRT; <span class="html-italic">n</span> = 5 tanks of 8 fish) groups across the two time intervals tested (from 0 to 5 and from 5 to 10 min for the shoaling test, and before and after the shoaling test for the cortisol concentration). Data are presented using box-and-whisker plots, indicating the range of the central 50% of the data, with a central line marking the median value. Lines extend from each box to capture the range of the remaining data (* <span class="html-italic">p</span> ≤ 0.05; ** <span class="html-italic">p</span> ≤ 0.01; **** <span class="html-italic">p</span> ≤ 0.0001).</p>
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18 pages, 1482 KiB  
Article
Contextual Factors Associated with Fecal Glucocorticoid Metabolites in Juvenile Polar Bears (Ursus maritimus) and a Cohabitating Juvenile Grizzly Bear (Ursus arctos horribilis) at the Detroit Zoo
by Emily Bovee, Tevon Madry, Kylen N. Gartland and Grace Fuller
J. Zool. Bot. Gard. 2025, 6(1), 1; https://doi.org/10.3390/jzbg6010001 - 9 Jan 2025
Viewed by 257
Abstract
Fecal glucocorticoid metabolites have been used to evaluate responses to stressors in captive adult polar (Ursus maritimus) and grizzly (Ursus arctos horribilis) bears. However, there is a lack of physiological information on juvenile bears in captivity that could help [...] Read more.
Fecal glucocorticoid metabolites have been used to evaluate responses to stressors in captive adult polar (Ursus maritimus) and grizzly (Ursus arctos horribilis) bears. However, there is a lack of physiological information on juvenile bears in captivity that could help expand the current understanding of their development and welfare. To address these questions, we tracked fecal glucocorticoid metabolites (FGMs) and behavior for 15 months in two polar bear cubs born at the Detroit Zoo, one who was mother-reared (Astra) and one who was hand-reared (Laerke), and one rescued grizzly bear cub (Jeb) reared at the Zoo. To allow access to a social partner during key developmental stages, Laerke and Jeb were housed together for eight months. Daily opportunistic samples were analyzed for fecal cortisol metabolites using an enzyme immunoassay and compared against behavior, social proximity, and environmental data gathered from 15 min focal observations. Based on a combination of generalized linear mixed models and Wilcoxon and Kruskal–Wallis tests, we found no significant variation in mean FGMs between Astra and Laerke, but both had significantly different mean FGMs compared to Jeb. We found that Laerke had higher FGM concentrations when she spent more time engaged in all-occurrence social negative behaviors and lower FGMs when engaged in social positive behaviors. For Jeb, FGMs were lower when in social proximity and higher following separation from Laerke. These data provide novel insights into the physiological states of juvenile bears during key stages and contribute to the growing body of information on polar and grizzly bear development. Full article
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<p>Social conditions for all three bear cubs.</p>
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<p>FGM concentrations in ng/g before and after a social separation in polar bears.</p>
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<p>FGM concentrations in ng/g for Jeb before and after the separation from Laerke.</p>
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<p>Longitudinal fecal glucocorticoid metabolite profile for Laerke. The top graph represents the combined Full Dosage conditions. The bottom graph represents the combined Decreasing Dosages conditions. FGM concentrations are expressed in ng/g of dry feces.</p>
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<p>FGM concentration (Mean ± SE) for Laerke and Jeb between housing conditions.</p>
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<p>Longitudinal fecal glucocorticoid metabolite profile for Jeb. FGM concentrations are expressed in ng/g of dry feces.</p>
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<p>Longitudinal fecal glucocorticoid metabolite profile for Astra. FGM concentrations are expressed in ng/g of dry feces.</p>
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13 pages, 1361 KiB  
Article
Analysis of the Reliability of Feather Sections for Corticosterone Measurement in Pekin Ducks
by Se-Jin Lim, Chan Ho Kim, Ka Young Yang, Woo Do Lee, Su Mi Kim, Yang-Ho Choi and Jung Hwan Jeon
Animals 2025, 15(2), 138; https://doi.org/10.3390/ani15020138 - 8 Jan 2025
Viewed by 331
Abstract
The aim of this study was to identify the feather section, among the whole feather, rachis, and vane, with the highest reliability for corticosterone measurement in 10 Pekin ducks aged 42 days. In total, 60 samples (i.e., 20 per section) were collected and [...] Read more.
The aim of this study was to identify the feather section, among the whole feather, rachis, and vane, with the highest reliability for corticosterone measurement in 10 Pekin ducks aged 42 days. In total, 60 samples (i.e., 20 per section) were collected and each section was analyzed in duplicate. Corticosterone levels were measured using ELISA and statistical analyses were performed using ANOVA in SAS 9.2, and the intra-class correlation coefficient (ICC) was tested using IBM SPSS. The level in the whole feather (12.55 ± 4.41 pg/mg) was significantly lower (p < 0.001) than that in the rachis (18.12 ± 5.70 pg/mg). No significant differences were observed between the rachis and the vane (20.40 ± 3.04 pg/mg). ANOVA results confirmed substantial hormonal variability depending on the feather part analyzed. The ICCs for the whole feather, rachis, and vane were 0.923, 0.876, and 0.004, respectively. The vane section exhibited the highest concentration and lowest variance, whereas the whole feather exhibited the highest ICC. Although the whole feather had the lowest concentration, it exhibited greater consistency. Further research is necessary to improve the feather corticosterone analysis method for more accurate analysis. In conclusion, the whole feather provides the most reliable measure of corticosterone concentration among the three parts. Full article
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Figure 1
<p>Photograph of feather samples showing the processing steps. (<b>a</b>) Feather sampling site (scapular region), (<b>b</b>) before mincing the whole feather, (<b>c</b>) after mincing the whole feather, (<b>d</b>) before mincing the rachis section, (<b>e</b>) after mincing the rachis section, (<b>f</b>) before mincing the vane section, (<b>g</b>) after mincing the vane section, (<b>h</b>) before pulverization, and (<b>i</b>) after pulverization.</p>
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<p>Bland–altman plot of corticosterone in the whole feather part. Red points represent the differences between the two measurements taken from two feathers per individual, plotted against their mean (<span class="html-italic">n</span> = 10). The solid horizontal line indicates the mean difference between the two measurements. The dashed horizontal lines represent the 95% limits of agreement, calculated as the mean difference ± 1.96 times the standard deviation of the differences.</p>
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<p>Corticosterone concentration (pg/mg) of the whole feather, rachis, and vane. Each point (circle, square, or triangle) represents an individual measurement of the respective feather parts. The horizontal line within each box indicates the median concentration. Boxes represent the interquartile range (IQR), and whiskers denote the data range, excluding outliers. a,b values above the box plot indicate a statistically significant difference (Tukey’s HSD test, p &lt; 0.05).</p>
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<p>Bland–altman plot of corticosterone in rachis part. Red points represent the differences between the two measurements taken from two feathers per individual, plotted against their mean (<span class="html-italic">n</span> = 10). The solid horizontal line indicates the mean difference between two measurements. The dashed horizontal lines represent the 95% limits of agreement, calculated as the mean difference ± 1.96 times the standard deviation of the differences.</p>
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<p>Bland–altman plot of corticosterone in vane part. Red points represent the differences between the two measurements taken from two feathers per individual, plotted against their mean (<span class="html-italic">n</span> = 10). The solid horizontal line indicates the mean difference between two measurements. The dashed horizontal lines represent the 95% limits of agreement, calculated as the mean difference ± 1.96 times the standard deviation of the differences.</p>
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16 pages, 2642 KiB  
Article
Enhancing Environmental Control in Broiler Production: Retrieval-Augmented Generation for Improved Decision-Making with Large Language Models
by Marcus Vinicius Leite, Jair Minoro Abe, Marcos Leandro Hoffmann Souza and Irenilza de Alencar Nääs
AgriEngineering 2025, 7(1), 12; https://doi.org/10.3390/agriengineering7010012 - 8 Jan 2025
Viewed by 226
Abstract
The growing global demand for animal protein, particularly chicken meat, challenges poultry farming to adapt production systems through the adoption of digital technologies. Among the promising advances in artificial intelligence (AI), large language models (LLMs) hold potential to enhance decision-making in broiler production [...] Read more.
The growing global demand for animal protein, particularly chicken meat, challenges poultry farming to adapt production systems through the adoption of digital technologies. Among the promising advances in artificial intelligence (AI), large language models (LLMs) hold potential to enhance decision-making in broiler production by supporting environmental control through the interpretation of climatic data, the generation of reports to optimize conditions, guidance on ventilation adjustments, recommendations for thermal management, assistance in air quality monitoring, and the translation of simulation results into actionable suggestions to improve bird welfare. For this purpose, the key limitations of LLMs in terms of transparency, accuracy, precision, and relevance must be effectively addressed. This study investigates the impact of retrieval-augmented generation (RAG) on improving LLM precision and relevance for environmental control in broiler production. Experiments with the OpenAI GPT-4o model and semantic similarity analysis were used to evaluate response quality with and without RAG. The results confirmed the approach’s effectiveness while identifying areas for improvement. A paired t-test revealed significantly higher similarity scores with RAG, demonstrating its impact on response quality. This study contributes to the field by advancing RAG-enhanced LLMs for environmental control, addressing market demands by demonstrating how AI improves decision-making for productivity and animal welfare, and benefits society by providing small-scale producers with cost-effective and accessible solutions for actionable insights. Full article
(This article belongs to the Special Issue The Future of Artificial Intelligence in Agriculture)
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Graphical abstract
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<p>Schematic of the RAG Process Flow. Source: the authors.</p>
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<p>Schematic flow of the research approach. Source: the authors.</p>
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<p>GPT-4o and RAG implementation user print screen interface. Source: the authors.</p>
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<p>Similarity index comparison, without RAG vs. with RAG. Source: the authors.</p>
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<p>Differences between the similarity rate with RAG and without RAG. Source: the authors.</p>
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<p>Differences between the frequencies of similarity indices with RAG and without RAG. Source: the authors.</p>
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<p>Boxplot comparison of the distributions considering the similarity indices with and without RAG. Source: the authors.</p>
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12 pages, 237 KiB  
Article
Beliefs About Cats and Dogs Among Pet Owners and Former Owners
by Tiffani J. Howell, Silvana Diverio and David J. Menor-Campos
Pets 2025, 2(1), 2; https://doi.org/10.3390/pets2010002 - 8 Jan 2025
Viewed by 294
Abstract
Despite an increase in research into cat and dog cognition, behavior, and welfare in recent decades, it is unclear whether pet owners are aware of recent advances in our understanding of cats and dogs. Misunderstandings about the reasons for animal behavior can lead [...] Read more.
Despite an increase in research into cat and dog cognition, behavior, and welfare in recent decades, it is unclear whether pet owners are aware of recent advances in our understanding of cats and dogs. Misunderstandings about the reasons for animal behavior can lead to negative welfare outcomes for the animal, and potentially distress for the owner, so it is important for owners to understand the latest science on animal behavior. Current and former pet owners (N = 224) completed an online survey rating their agreement with a series of statements about cat and dog behavior. After completing the survey, participants were provided with a document describing the latest scientific knowledge about each of the statements in the survey. For both species, participant beliefs generally accord with the latest scientific knowledge, but there is evidence of remaining beliefs about aspects of dominance training theory in dogs, and the belief that cats are low-maintenance pets. These findings can be used by trainers and behaviorists to help educate owners about pet needs, working from the owner’s baseline knowledge and debunking persistent myths. Full article
11 pages, 1612 KiB  
Article
Wild Boar Attacks on Hunting Dogs in Czechia: The Length of the Hunting Season Matters
by Jana Adámková, Karolína Lazárková, Jan Cukor, Hana Brinkeová, Jitka Bartošová, Luděk Bartoš and Kateřina Benediktová
Animals 2025, 15(2), 130; https://doi.org/10.3390/ani15020130 - 8 Jan 2025
Viewed by 244
Abstract
Hunting dogs are exposed to the risk of injury in driven hunts, an often-used method for managing growing wild boar numbers. This study investigated the impact of increased hunting pressure—both across the hunting season and within individual hunting events—on the risk of wild [...] Read more.
Hunting dogs are exposed to the risk of injury in driven hunts, an often-used method for managing growing wild boar numbers. This study investigated the impact of increased hunting pressure—both across the hunting season and within individual hunting events—on the risk of wild boar attacks on hunting dogs, i.e., the length of the hunting season (2.68 ± 0.76 months, mean ± standard deviation), the number of driven hunts per season (3.99 ± 0.43), the intervals between hunts (17.85 ± 4.83 days), the number of wild boars harvested per season (14.46 ± 13.10), and the number of participants (23.8 ± 10.69) and dogs (4.56 ± 2.66) involved per hunt. The data were collected via a retrospective questionnaire survey. The information-theoretic approach (IT-AIC) and GLMM were employed to estimate the factors’ effects on the number of wild boar attacks on dogs reported in 40 hunting grounds in five consecutive hunting seasons (2.60 ± 5.07 attacks per hunting season in a hunting ground). The number of attacks only increased with the length of the hunting season. The best model did not include other factors, such as shorter intervals between hunts, a higher number of driven hunts, wild boars harvested, or participants. The respondents reported 150 injuries by wild boars during 797 driven hunts. Most injuries were mild (73.8%), with fewer severe (18.8%) and fatal (7.4%) cases. Further investigation into wild boar and hunting dog interactions is necessary for constructing strategies to improve hunting practices and reduce dog injury risks. Full article
(This article belongs to the Section Wildlife)
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<p>The study area map shows 40 hunting grounds (game management areas) with valid responses and the distribution of the forested regions and agricultural fields. EN: © EuroGeographics for the administrative boundaries. OpenStreetMap. Source service: © CENIA, česká informační agentura životního prostředí, Source data: © Agentura ochrany přírody a krajiny, Available online: Národní geoportál INSPIRE <a href="http://geoportal.gov.cz" target="_blank">http://geoportal.gov.cz</a> (accessed on 27 December 2024).</p>
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<p>Predicted values of the number of wild boar attacks reported during a hunting season on dogs (log-transformed) with 95% confidence intervals according to the hunting season length (<span class="html-italic">x</span>-axis). The bubble size refers to the number of hunting seasons the data were obtained from (n = 195).</p>
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22 pages, 2144 KiB  
Article
Heart Rate Monitoring During Behavioral Stress Tests in Bold and Shy Rainbow Trout (Oncorhynchus mykiss)
by Eleftherios Kasiouras, Gautier Riberolles, Albin Gräns, Andreas Ekström, Johan Höjesjö, Jonathan A. C. Roques, Erik Sandblom and Lynne U. Sneddon
Fishes 2025, 10(1), 23; https://doi.org/10.3390/fishes10010023 - 7 Jan 2025
Viewed by 469
Abstract
Monitoring stress in captive fish is crucial for their welfare, but continuous physiological measures in unrestrained animals are challenging. Rainbow trout (Oncorhynchus mykiss) exhibit divergent personalities, ranging from bold to shy, which correlate with cortisol-mediated stress responses. To determine whether personality [...] Read more.
Monitoring stress in captive fish is crucial for their welfare, but continuous physiological measures in unrestrained animals are challenging. Rainbow trout (Oncorhynchus mykiss) exhibit divergent personalities, ranging from bold to shy, which correlate with cortisol-mediated stress responses. To determine whether personality affects the sympathetic nervous system, heart rate was measured during three potentially stressful events as a proxy for sympathetic nervous system responses. Firstly, trout were classified as bold or shy, using a novel object test. Subsequently, trout were implanted with biologgers to record heart rate in vivo at rest during and after the behavioral tests. Following recovery, the fish underwent a second novel object test, a confinement test, a pair-wise contest, and a final novel object test to explore the degree of boldness over the experimental period, which remained consistent. Heart rate was relatively higher in both bold and shy animals during the confinement test and the pair-wise contest compared with the novel object test, which indicated that heart rate monitoring was a valid gauge of the valence of the experience. Heart rate responses did not differ between bold and shy trout, indicating that behavioral phenotype did not influence the autonomic stress response. Thus, heart rate is a reliable indicator of stress without the need to account for intra-specific behavioral variations. Full article
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Graphical abstract
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<p>(<b>a</b>) Mean (±SE) time that fish spent in the 15,10, 5 cm zone (s), between bold and shy rainbow trout across the three novel object tests. (<b>b</b>) Mean (±SE) of the frequency that fish entered the 15, 10, 5 cm zone, between bold and shy rainbow trout across the three novel object tests. (<b>c</b>) Mean (±SE) time that fish spent in greater distance than 15 cm and inactivity time (s), between bold and shy rainbow trout across the three novel object tests (n = 16, bold; n = 16, shy; * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.001).</p>
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<p>Mean (±SE) of the dominance index during pair-wise contests in rainbow trout (n = 16, bold; n = 16, shy; * <span class="html-italic">p</span> &lt; 0.05).</p>
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<p>(<b>a</b>) Mean (±SE) of the minimum value of the heart rate (HR, in beats per min, bpm) during the novel object test, confinement test and pair-wise contest in rainbow trout (n = 16, bold; n = 16, shy). (<b>b</b>) Mean (±SE) of the maximum value of the heart rate during the novel object test, confinement test and pair-wise contest (n = 16, bold; n = 16, shy; * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01). No significant differences were found between bold and shy animals.</p>
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<p>(<b>a</b>) Mean (±SE) of the minimum value of the heart rate (HR, in beats per min, bpm) during the novel object test, confinement test and pair-wise contest in rainbow trout (n = 16, bold; n = 16, shy). (<b>b</b>) Mean (±SE) of the maximum value of the heart rate during the novel object test, confinement test and pair-wise contest (n = 16, bold; n = 16, shy; * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01). No significant differences were found between bold and shy animals.</p>
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<p>(<b>a</b>) Mean (±SE) of the average resting heart rate (HR, in beats per min, bpm) before the three tests (novel object test, confinement test, and pair-wise contest) between bold and shy rainbow trout (n = 16, bold; n = 16, shy). (<b>b</b>) Mean (±SE) of the average heart rate during the three tests (novel object test, confinement test, and pair-wise contest) between bold and shy (n = 16, bold; n = 16, shy). (<b>c</b>) Mean (±SE) of the average resting heart rate after the three tests (novel object test, confinement test, and pair-wise contest) between bold and shy (n = 16, bold; n = 16, shy; *** <span class="html-italic">p</span> &lt; 0.001). No significant differences were found between bold and shy animals.</p>
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<p>Mean (±SE) of the 20th percentile of recovery rate between the three competitions (novel object test, confinement test, and pair-wise contest between bold and shy rainbow trout (n = 16, bold; n = 16, shy; * <span class="html-italic">p</span> &lt; 0.05). The 20th percentile of the recovery rate is defined as the 20th percentile of all measurements during the test divided by the recovery time. No significant differences were found between bold and shy animals.</p>
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<p>Mean (±SE) of the recovery time between the three competitions (novel object test, confinement test, and pair-wise contest) between bold and shy rainbow trout (n = 16, bold; n = 16, shy; *** <span class="html-italic">p</span> &lt; 0.001). No significant differences were found between bold and shy animals.</p>
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<p>Pearson correlation between the absolute values of latency to approach a novel object to within 5 cm and the absolute values of the average heart rate (HR) before the 2nd novel object test in rainbow trout.</p>
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<p>Spearman correlation between the absolute values of dominance index and the absolute values of average heart rate (HR) before the pair-wise contest in rainbow trout.</p>
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12 pages, 249 KiB  
Article
Behavioral and Physiological Responses of Therapy Dogs to Animal-Assisted Treatment in an Inpatient Stroke Rehabilitation Program
by Hao-Yu Shih, François Martin, Debra Ness, Whitney Romine, Taylor L. Peck, Tricia Turpin, Rachael Horoschak, Cindy Steeby, Hannah Phillips, Mary Claypool, Amanda Theuer, Grace M. Herbeck, Jasmine Sexton, Erin Pittman, Erica Bellamkonda, Nikita Maria Ligutam Mohabbat, Sandra A. Lyn, Brent A. Bauer and Arya B. Mohabbat
Animals 2025, 15(2), 121; https://doi.org/10.3390/ani15020121 - 7 Jan 2025
Viewed by 618
Abstract
Therapy dogs have been increasingly incorporated into a variety of medical treatment programs to improve patients’ treatment outcomes and wellbeing. However, research investigating the stress level of therapy dogs in this setting is limited. This is the first randomized–controlled and prospective study that [...] Read more.
Therapy dogs have been increasingly incorporated into a variety of medical treatment programs to improve patients’ treatment outcomes and wellbeing. However, research investigating the stress level of therapy dogs in this setting is limited. This is the first randomized–controlled and prospective study that investigated the wellbeing of therapy dogs in an inpatient stroke rehabilitation program. In this study, 14 therapy dog–handler pairs were embedded in an inpatient stroke rehabilitation program to provide animal-assisted treatment (AAT). These therapy dog–handler pairs actively participated in stroke rehabilitation by walking with the patient, playing fetch with the patient, and being petted/brushed by the patient, amongst various other AAT activities. To measure canine stress responses during the rehabilitation sessions, salivary cortisol and oxytocin concentrations, heart rate and heart rate variability, tympanic membrane temperature, and a behavioral evaluation were recorded before and after interactions with the patient. The results demonstrated that therapy dogs had significantly decreased heart rate and increased heart rate variability after the AAT session. Right tympanic temperature significantly increased after the session, but there was no significant difference in terms of salivary cortisol or oxytocin levels, nor in stress-related behavioral evaluations after the AAT session. Taken together, the results suggest that incorporating AAT into an inpatient stroke rehabilitation program did not induce stress in the therapy dogs, and that the therapy dogs may have been more relaxed after the session. Full article
(This article belongs to the Special Issue The Complexity of the Human–Companion Animal Bond)
16 pages, 5868 KiB  
Article
A Deep Learning-Based Approach for Precise Emotion Recognition in Domestic Animals Using EfficientNetB5 Architecture
by Rashadul Islam Sumon, Haider Ali, Salma Akter, Shah Muhammad Imtiyaj Uddin, Md Ariful Islam Mozumder and Hee-Cheol Kim
Eng 2025, 6(1), 9; https://doi.org/10.3390/eng6010009 - 3 Jan 2025
Viewed by 430
Abstract
The perception of animal emotions is key to enhancing veterinary practice, human–animal interactions, and protecting domesticated species’ welfare. This study presents a unique emotion classification deep learning-based approach for pet animals. The actual and emotional status of dogs and cats have been classified [...] Read more.
The perception of animal emotions is key to enhancing veterinary practice, human–animal interactions, and protecting domesticated species’ welfare. This study presents a unique emotion classification deep learning-based approach for pet animals. The actual and emotional status of dogs and cats have been classified using a modified EfficientNetB5 model. Utilizing a dataset of images classified into four different emotion categories—angry, sad, happy, and neutral—the model incorporates sophisticated feature extraction methods, such as Dense Residual Blocks and Squeeze-and-Excitation (SE) blocks, to improve the focus on important emotional indicators. The basis of the second strategy is EfficientNetB5, which is known for providing an optimal balance in terms of accuracy and processing capabilities. The model exhibited robust generalization abilities for the subtle identification of emotional states, achieving 98.2% accuracy in training and 91.24% during validation on a separate dataset. These encouraging outcomes support the model’s promise for real-time emotion detection applications and demonstrate its adaptability for wider application in ongoing pet monitoring systems. The dataset will be enlarged, model performance will be enhanced for more species, and real-time capabilities will be developed for real-world implementation. Full article
(This article belongs to the Special Issue Artificial Intelligence for Engineering Applications)
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<p>Flow diagram of pet animal face emotion detection.</p>
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<p>Categorical images that show the four emotional states of domestic animals. Images in the first row are categorized as Angry, in the second row as Neutral/Other, in the third row as Sad, and in the fourth row as Happy.</p>
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<p>The left panel displays the original images, while the right panel displays the corresponding pre-processed images after applying the noise and blur reduction algorithms.</p>
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<p>Backbone architecture of EfficientNetB5 model for pet emotion classification.</p>
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<p>Squeeze-and-Excitation module architecture.</p>
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<p>Dense Residual Block architecture.</p>
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<p>Comparison training and testing accuracy of (<b>a</b>) Mobile Net, (<b>b</b>) VGG-16, (<b>c</b>) Inception V3, (<b>d</b>) Alex Net, (<b>e</b>) Exception, (<b>f</b>) Dense Net, (<b>g</b>) Res Net-50, and (<b>h</b>) Proposed Modified EfficientNetB5.</p>
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<p>Comparison confusion matrices of (<b>a</b>) Mobile Net, (<b>b</b>) VGG-16, (<b>c</b>) Inception V3, (<b>d</b>) Alex Net, (<b>e</b>) Exception, (<b>f</b>) Dense Net, (<b>g</b>) Res Net-50, and (<b>h</b>) Proposed Modified EfficientNetB5.</p>
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<p>Prediction results of proposed Modified EfficientNetB5 model.</p>
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9 pages, 1466 KiB  
Brief Report
Behavioral Signature of Equine Gastric Discomfort? Preliminary Retrospective Clinical Observations
by Catherine Torcivia and Sue M. McDonnell
Animals 2025, 15(1), 88; https://doi.org/10.3390/ani15010088 - 3 Jan 2025
Viewed by 1062
Abstract
Gastric ulcer disease and other potentially painful gastric conditions are among the most common afflictions adversely affecting the welfare of domestic equids. A large percentage of affected animals may not display the classic signs of gastric disease, such as unexplained weight loss, poor [...] Read more.
Gastric ulcer disease and other potentially painful gastric conditions are among the most common afflictions adversely affecting the welfare of domestic equids. A large percentage of affected animals may not display the classic signs of gastric disease, such as unexplained weight loss, poor hair coat, and inappetence until the disease becomes severe. As a clinical service within our equine referral hospital, we routinely evaluate 24-h video recorded samples of horses to assist clinicians in identifying subtle discomfort and potential sources or to scan for infrequent neurologic or cardiac-related behavioral events. Empirically, we have recognized discomfort behaviors that appear to be uniquely associated with gastric disease. These include frequent attention to the cranial abdomen (nuzzling, swatting, nipping, and/or caudal gaze focused on the abdomen caudal to the elbow) and/or deep abdominal stretching, often within the context of eating, drinking, and/or anticipating feeding. To systematically evaluate the reliability of these purported gastric discomfort behaviors, we reviewed 30 recent 24-h video behavior evaluation cases for which (1) the clinical video behavior evaluation had been carried out without knowledge of the history and presenting complaint and (2) direct gastric examination had confirmed gastric disease status at the time. Twenty-four of the thirty cases showed gastric discomfort behavior, and all twenty-four had either gastric ulcers (n = 21) and/or gastric impaction (n = 3). Of the six cases not showing gastric discomfort behaviors, four were free of gastric disease, while two had mild lesions. Comparing horses with and without gastric disease, gastric discomfort behaviors were reported in 24 of the 26 (92%) with gastric ulcers or gastric impaction, compared to none of the four gastric disease-free horses. Although a larger prospectively designed study is needed to confidently estimate the sensitivity and specificity or the associations of behavior with the type or severity of gastric disease, these results confirm our long-held clinical impression of a behavioral signature for gastric discomfort in the horse. Full article
(This article belongs to the Special Issue Focus on Gut Health in Horses: Current Research and Approaches)
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<p>Gastric discomfort behaviors.</p>
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14 pages, 1011 KiB  
Article
First Report of SNPs Detection in TMEM154 Gene in Sheep from Poland and Their Association with SRLV Infection Status
by Magdalena Materniak-Kornas, Katarzyna Piórkowska, Katarzyna Ropka-Molik, Adrianna Dominika Musiał, Joanna Kowalik, Anna Kycko and Jacek Kuźmak
Pathogens 2025, 14(1), 16; https://doi.org/10.3390/pathogens14010016 - 30 Dec 2024
Viewed by 426
Abstract
Small ruminant lentiviruses (SRLVs) infect sheep, causing a multiorganic disease called maedi-visna or ovine progressive pneumonia, which significantly affects the production and welfare of sheep, generating serious economic losses. Although not all infected animals develop fully symptomatic disease, they constantly spread the virus [...] Read more.
Small ruminant lentiviruses (SRLVs) infect sheep, causing a multiorganic disease called maedi-visna or ovine progressive pneumonia, which significantly affects the production and welfare of sheep, generating serious economic losses. Although not all infected animals develop fully symptomatic disease, they constantly spread the virus in the flock. Since the infection is incurable and no vaccine is available, another approach is necessary to control SRLV infections. In recent years, an alternative for culling infected animals has become the approach based on identifying genetic markers for selecting SRLV-resistant individuals. Recent reports revealed several candidates, including gene encoding transmembrane protein 154 (TMEM154). Several single nucleotide polymorphisms (SNPs) are found within this gene in sheep of different breeds, but only some can be considered as resistant markers. This study aimed to investigate the presence of single polymorphic sites in TMEM154 gene in sheep of selected Polish flocks and assess their association with the infection and proviral load in the context of susceptibility to SRLV infection. In total 107 sheep, representing three breeds, were screened for their SRLV infection status by serological and PCR testing. All these animals were also genotyped to characterize the presence of SNPs in TMEM154 gene and estimate their potential of being the SRLV-resistance marker. The frequency of identified alleles differed among breeds. Moreover, the positive association between TMEM154 genotype and SRLV status was found for E35K polymorphism and two polymorphic sites in 5′UTR in one of analyzed breed. However, when the relationship between SNPs and SRLV proviral load was analyzed, five had a strong association, considering the whole population of tested sheep. Presented data, for the first time, identified the presence of SNPs in TMEM154 gene in sheep housed in Polish flocks and suggested that selecting SRLV-resistant animals based on this analysis might be possible, but further validation in a larger group of sheep is required. Full article
(This article belongs to the Section Viral Pathogens)
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<p>Distribution of the SRLV copy numbers in sheep from three breeds. The grey dots present the provirus copy number of each sheep, the black dashed line represents the mean of the SRLV copy number in each breed, and the orange error bars represent the 95% CI for the mean.</p>
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<p>The association of TMEM154 genotypes with SRLV proviral load.</p>
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<p>The association of selected TMEM154 diplotypes (according to <a href="#app1-pathogens-14-00016" class="html-app">Table S3</a>) with SRLV proviral load.</p>
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13 pages, 186 KiB  
Commentary
An Overview of Greece’s Newly Established Progressive Stray Dog Management Laws
by Christie Siettou, Eleni Theodoropoulou and Anna Stefani Siettou
Pets 2025, 2(1), 1; https://doi.org/10.3390/pets2010001 - 29 Dec 2024
Viewed by 470
Abstract
This study provides an overview of the new Greek legislative framework and highlights its progressive nature in tackling one of the most populous stray animal populations in the world. We present the newly established law and discuss its provisions, aims, and challenges. We [...] Read more.
This study provides an overview of the new Greek legislative framework and highlights its progressive nature in tackling one of the most populous stray animal populations in the world. We present the newly established law and discuss its provisions, aims, and challenges. We also examine the reception of the law by key stakeholders such as veterinary practitioners, animal welfare organisations, and the Hellenic Kennel Club. With the post-implementation review scheduled for 2026, its evaluation has yet to be conducted. Full article
30 pages, 3408 KiB  
Article
Social Relationships of Captive Bachelor Przewalski’s Horses and Their Effect on Daily Activity and Space Use
by Anastasiia Nykonenko, Yevhen Moturnak and Philip Dunstan McLoughlin
Animals 2025, 15(1), 53; https://doi.org/10.3390/ani15010053 - 28 Dec 2024
Viewed by 654
Abstract
Understanding social relationships in at-risk species held in captivity is vital for their welfare and potential reintroduction. In social species like the Przewalski’s horse (Equus ferus przewalskii), daily time allocation and space use may be influenced by social structure and, in [...] Read more.
Understanding social relationships in at-risk species held in captivity is vital for their welfare and potential reintroduction. In social species like the Przewalski’s horse (Equus ferus przewalskii), daily time allocation and space use may be influenced by social structure and, in turn, reflect welfare. Here, we identify social relationships, time budgets, and spatial distribution of a group of nine older (aged 6–21 years) male Przewalski’s horses living in a non-breeding (bachelor) group. We conducted our work at the Askania-Nova Biosphere Reserve, over 65 h of observation in summer, 2015. Horses formed stronger social bonds with individuals of similar gregariousness and dominance levels. Social-network analysis identified three distinct subgroups with significant differences in locomotion, social behaviour, and foraging. However, resting and vigilance behaviour remained similar across the subgroups. Behavioural synchrony across all activities was higher within subgroups than between. Space use was also affected by subgroup membership, with some horses overusing or underusing critical resource areas like hay and water. These findings suggest opportunities to improve welfare by adjusting space and resource distribution, particularly for more submissive individuals. Our approach may also aid in selecting reintroduction candidates by considering social characteristics alongside health, genetic, and other factors. Full article
(This article belongs to the Special Issue The Relevance of Companionship and Social Behaviour for Horses)
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<p>Enclosure map (I, II) for adult male Przewalski’s horses, Askania-Nova Biosphere Reserve (21 July to 14 August 2015).</p>
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<p>The social network of association indices in the bachelor group of Przewalski’s horses (Social cluster = communities detected by Louvain method).</p>
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<p>The network of rates of affiliative interactions in a bachelor group of Przewalski’s horses (arrow direction reflects initiator and a recipient of affiliative behaviour; Social cluster = communities detected by Infomap method).</p>
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<p>The relationships between weighted degree and strength derived from association network (<b>left</b>) and network of affiliative interaction rates (<b>right</b>) of Przewalski’s horses males.</p>
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<p>The network of rates of agonistic interactions in a bachelor group of Przewalski’s horses (arrow direction reflects initiator and a recipient of either offensive or defensive behaviour; weight = rates of interactions, clusters = communities detected by Infomap method).</p>
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<p>Daily time budget of Przewalski’s bachelor males by time of day (mean percentage of behaviour ± s.d.).</p>
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<p>Behavioural synchrony in the bachelor group of Przewalski’s horses by activities by social clusters (see <a href="#animals-15-00053-t008" class="html-table">Table 8</a>).</p>
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<p>Visualization of daily time budget of a bachelor group of Przewalski’s horses with different enclosure availability (values for all activities for all study period).</p>
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<p>Electivity indices by zone with different resource availability for horses in periods with two and one enclosure available.</p>
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<p>Activities performed by Przewalski’s horses in different zones of the enclosures.</p>
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16 pages, 3816 KiB  
Article
Automated Dead Chicken Detection in Poultry Farms Using Knowledge Distillation and Vision Transformers
by Ridip Khanal, Wenqin Wu and Joonwhoan Lee
Appl. Sci. 2025, 15(1), 136; https://doi.org/10.3390/app15010136 - 27 Dec 2024
Viewed by 433
Abstract
Detecting dead chickens in broiler farms is critical for maintaining animal welfare and preventing disease outbreaks. This study presents an automated system that leverages CCTV footage to detect dead chickens, utilizing a two-step approach to improve detection accuracy and efficiency. First, stationary regions [...] Read more.
Detecting dead chickens in broiler farms is critical for maintaining animal welfare and preventing disease outbreaks. This study presents an automated system that leverages CCTV footage to detect dead chickens, utilizing a two-step approach to improve detection accuracy and efficiency. First, stationary regions in the footage—likely representing dead chickens—are identified. Then, a deep learning classifier, enhanced through knowledge distillation, confirms whether the detected stationary object is indeed a chicken. EfficientNet-B0 is employed as the teacher model, while DeiT-Tiny functions as the student model, balancing high accuracy and computational efficiency. A dynamic frame selection strategy optimizes resource usage by adjusting monitoring intervals based on the chickens’ age, ensuring real-time performance in resource-constrained environments. This method addresses key challenges such as the lack of explicit annotations for dead chickens, along with common farm issues like lighting variations, occlusions, cluttered backgrounds, chicken growth, and camera distortions. The experimental results demonstrate validation accuracies of 99.3% for the teacher model and 98.7% for the student model, with significant reductions in computational demands. The system’s robustness and scalability make it suitable for large-scale farm deployment, minimizing the need for labor-intensive manual inspections. Future work will explore integrating deep learning methods that incorporate temporal attention mechanisms and automated removal processes. Full article
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<p>The pipeline for the two-stage dead chicken detection.</p>
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<p>Stationary object detection process.</p>
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<p>Chart and block diagram showing the frame selection rate across the age of the chicken (<b>a</b>,<b>b</b>). In (<b>b</b>), frames from the CCTV footage are represented in blue and orange colors. The orange color represents the captured frames for evaluation.</p>
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<p>Training and validation loss and accuracy curves: (<b>a</b>,<b>b</b>) are the training and validation loss curves for teacher and student models, respectively. (<b>c</b>,<b>d</b>) are the training and validation accuracy curves for both models.</p>
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<p>Confusion matrix for teacher and student model on the validation dataset.</p>
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<p>Bar chart comparing inference time and model size.</p>
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<p>Examples of correct detections in (<b>a</b>,<b>b</b>) and common failure cases in (<b>c</b>). The first column contains the images of stationary object detection (within green box), and the second column contains the images of the confirmed dead chickens (within red box).</p>
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