Animals

16 pages, 2194 KB

Open AccessArticle

Metal Levels in Crab Sentinel Species from the Mediterranean Sea: Safety and Toxicological Risk Assessment

by Clara Naccari, Gaetano Cammilleri, Davide Alfonso Cammarata, Roberta Cicala, Antonio Procopio, Vincenzo Ferrantelli and Ernesto Palma

Animals 2026, 16(5), 857; https://doi.org/10.3390/ani16050857 - 9 Mar 2026

Viewed by 628

Abstract

The assessment of metal contamination in aquatic species represents an important tool to monitor the risk of exposure for these species in relation to marine pollution. Crabs are the sentinel species of the aquatic environment and are very sensitive to metal exposure because [...] Read more.

The assessment of metal contamination in aquatic species represents an important tool to monitor the risk of exposure for these species in relation to marine pollution. Crabs are the sentinel species of the aquatic environment and are very sensitive to metal exposure because these inorganic contaminants can be easily absorbed into their exoskeleton and migrate into their inner tissues. The aim of this study was to assess the content of metals and metalloids (Hg, Pb, Cd, As, Cr, Mn and Ni) in species of warty crab (Eriphia verrucosa), brown crab (Cancer pagurus), and marbled crab (Pachygrapsus marmoratus), mainly distributed along the Mediterranean coastlines. The investigations, carried out on several parts of crabs (carapace and the related pulp, gills, claws and bronchial muscle) showed the presence of residual levels of metals and metalloids (Mn > Pb > Hg > As > Ni > Cr > Cd), correlated to the distribution of these elements in the marine environment and to the specific biological features of crabs. However, the values of Metal Pollution Index (MPI) suggested a low level of contamination of the marine ecosystem where these species were found, and the coefficient of correlation (K), a specific parameters of toxicological risk assessment, was indicative of crab health status. However, the residual levels of Hg, Pb and As above the MRL in all analyzed crab species underlined the importance of continuous monitoring studies on the safety of these sea-foods. Full article

(This article belongs to the Special Issue Sentinel Species and Environmental Pollutants Exposure in Animal Health)

► Show Figures

Figure 1

22 pages, 3974 KB

Open AccessArticle

Climate-Driven Variation in Yellowfin Tuna Productivity in the Western and Central Pacific Ocean Inferred from a State-Space Model

by Xiaodong Li, Zhe Geng, Jie Cao, Jizhang Zhu and Jiangfeng Zhu

Animals 2026, 16(5), 856; https://doi.org/10.3390/ani16050856 - 9 Mar 2026

Viewed by 651

Abstract

Understanding temporal variation in population productivity is critical for effective assessment and management of pelagic fish stocks under a changing climate. In this study, we applied a stochastic surplus production model in continuous time (SPiCT) with time-varying parameters to evaluate the productivity dynamics [...] Read more.

Understanding temporal variation in population productivity is critical for effective assessment and management of pelagic fish stocks under a changing climate. In this study, we applied a stochastic surplus production model in continuous time (SPiCT) with time-varying parameters to evaluate the productivity dynamics of yellowfin tuna (Thunnus albacares) in the western and central Pacific Ocean and to examine the influence of environmental variability on productivity. Multiple time-varying parameterization scenarios were explored to characterize uncertainties in productivity estimates and associated biological reference points. Generalized additive models were subsequently used to quantify the relationships between environmental variables and time-varying productivity. Results indicate that productivity estimates exhibit consistent temporal patterns across alternative modeling scenarios, while their magnitude and associated uncertainty are sensitive to model structure. Among the environmental factors examined, the Pacific Decadal Oscillation (PDO) and mixed layer thickness (MLT) showed consistent and statistically significant associations with maximum net productivity. Higher PDO values and greater MLT were both positively associated with population productivity. Overall, the results highlight the importance of environmental variability in shaping time-varying productivity of yellowfin tuna and demonstrate the feasibility of incorporating key environmental indicators into a state-space model. This approach provides a complementary framework for interpreting stock dynamics and supports the development of ecosystem-based fisheries management strategies in the western and central Pacific. Full article

(This article belongs to the Special Issue Research on Fish Population Dynamics)

► Show Figures

Figure 1

18 pages, 8878 KB

Open AccessArticle

Canine Adipose MSC-Derived Exosomes Ameliorate Skeletal Muscle Injury in Mice

by Jiaxuan Gao, Yujue Li and Yougang Zhong

Animals 2026, 16(5), 855; https://doi.org/10.3390/ani16050855 - 9 Mar 2026

Viewed by 538

Abstract

Severe skeletal muscle injury in dogs can result in muscle atrophy, fibrotic remodeling, and fat accumulation, leading to skeletal muscle dysfunction and impaired quality of life. However, there is currently no effective treatment available. This study aims to investigate the potential of canine [...] Read more.

Severe skeletal muscle injury in dogs can result in muscle atrophy, fibrotic remodeling, and fat accumulation, leading to skeletal muscle dysfunction and impaired quality of life. However, there is currently no effective treatment available. This study aims to investigate the potential of canine adipose mesenchymal stem cell-derived exosomes (cADMSC-Exos) as a novel acellular therapy for the repair of muscle atrophy and injury. cADMSCs and their derived exosomes were isolated and characterized. A dexamethasone-induced C2C12 myotube atrophy model was established to evaluate the effects of cADMSC-Exos on muscle atrophy by assessing myotube morphology and the expression of atrophy-related factors. Subsequently, a glycerol-induced mouse muscle injury model was constructed. Through histological analysis and Western blot, the efficacy and safety of cADMSC-Exos in vivo were systematically evaluated. Results indicated that cADMSC-Exos demonstrated significant anti-atrophic activity in both two models, ameliorating skeletal muscle atrophy and the upregulation of muscle RING finger 1 (MuRF1) and muscle atrophy F-box (Atrogin-1) (p < 0.05), consistent with morphological alterations. Moreover, cADMSC-Exos markedly alleviated fibrosis and fatty infiltration in injured muscle tissue (p < 0.0001). Overall, these findings indicate that cADMSC-Exos promote muscle repair and attenuate pathological remodeling by modulating the local microenvironment and protein expression, highlighting their potential as a therapeutic strategy for muscular disorders. Full article

(This article belongs to the Special Issue Causes, Symptoms and Treatments of Musculoskeletal Diseases in Dogs and Cats)

► Show Figures

Figure 1

34 pages, 3096 KB

Open AccessReview

A Comparative Review of Fertility and Semen Assessment Techniques in Farm Animals

by Nada N. A. M. Hassanine, Nasir A. Ibrahim, Nosiba S. Basher, Ahmed A. Saleh, Shaaban S. Elnesr, Mohamed Osman Abdalrahem Essa, Hosameldeen Mohamed Husien and Mengzhi Wang

Animals 2026, 16(5), 854; https://doi.org/10.3390/ani16050854 - 9 Mar 2026

Cited by 1 | Viewed by 2374

Abstract

This review provides a comparative examination of current methods for evaluating fertility and semen quality in mammals, with emphasis on livestock species. It is based on a structured search and analysis of 316 academic publications retrieved from major scientific databases, including Elsevier, Springer, [...] Read more.

This review provides a comparative examination of current methods for evaluating fertility and semen quality in mammals, with emphasis on livestock species. It is based on a structured search and analysis of 316 academic publications retrieved from major scientific databases, including Elsevier, Springer, Taylor & Francis, and MDPI, as well as other repositories, covering literature published between 1938 and 2025. This review examines advanced semen analysis techniques, including computer-assisted semen analysis (CASA) systems and assays for assessing chromatin integrity, DNA damage, seminal plasma composition, sperm membrane proteins, and functional parameters such as migration capacity, membrane integrity, acrosomal status, capacitation, morphology, viability, concentration, motility, and volume. Methods for semen collection are also outlined. The synthesis focuses on the comparative assessment of techniques and recent findings for major ruminant and monogastric livestock species: cattle (Bos taurus and Bos indicus), buffalo (Bubalus bubalis), sheep (Ovis aries), goats (Capra hircus), and pigs (Sus domesticus). A brief historical overview of assisted insemination is presented, incorporating relevant procedures and managerial factors that affect fertility outcomes. Advances in genetics related to reproductive health are also discussed. This synthesis seeks to integrate existing knowledge and guide future research in reproductive science for both animal production and human medicine. Full article

(This article belongs to the Special Issue Livestock Fertility and Artificial Insemination)

► Show Figures

Figure 1

19 pages, 1572 KB

Open AccessArticle

Feeding an Essential Oil Blend to Growing Crossbred Lambs Mitigates Heat Stress to Improve Growth Performance via Enhanced Antioxidant Capacity

by Yannan Ma, Lei Yang, Fan Wu, Jiao Luo, Zhixian Liu, Wen Chen, Zhaomin Lei, Pengjia He, Ting Liu, Shuzhen Song, Shuai Wang, Jianping Wu and David P. Casper

Animals 2026, 16(5), 853; https://doi.org/10.3390/ani16050853 - 9 Mar 2026

Viewed by 544

Abstract

Heat stress (HS) poses a significant stress challenge to growing lambs, impacting growth performance, health, and physiological responses. The study evaluated feeding an essential oil blend (EOB) on growth performance, physiological and blood parameters, oxidative, and immune responses. Thirty-two 3 mo old female [...] Read more.

Heat stress (HS) poses a significant stress challenge to growing lambs, impacting growth performance, health, and physiological responses. The study evaluated feeding an essential oil blend (EOB) on growth performance, physiological and blood parameters, oxidative, and immune responses. Thirty-two 3 mo old female (BW 18.6 ± 2.43 kg) crossbred (Mongolian × Thin-tailed Han F1) lambs were randomly assigned to either: (1) control (CON)—grain mix without EOB; or (2) EOB—an EOB blend supplemented at 4 g/kg grain mix. The EOB blend was comprised of 4.34% Zanthoxylum, 1% capsicum, and 1.06% cinnamon oils with 93.6% attapulgite carrier containing linalool, sabinene, limonene, capsaicin, cinnamaldehyde and eugenol. Extreme heat stress occurred for 39 out of the 45 d experiment. Feeding HS lambs EOB increased (p < 0.05) ADG (107.4 and 162.0 g/d for CON and EOB, respectively), forage intake (239.2 and 287.0 g/d), DMI (863.1 and 930.2 g/d), and feed efficiency (0.123 and 0.181 ADG, g/DMI, g) compared with lambs fed CON. Feeding EOB reduced (p < 0.01) respiration rates (RR) and rectal (RT) and skin temperatures (ST) compared to CON-fed lambs. Lambs fed EOB had a higher T-SOD activity and IgM concentration (0.05 < p < 0.10) than CON-fed lambs. Growing heat-stressed lambs fed EOB demonstrated a lower (p < 0.05) eosinophil percentage. Feeding EOB ameliorated HS conditions for growing crossbred lambs, which demonstrate improved growth performance, enhanced physiological responses, and overall health status. Full article

(This article belongs to the Section Animal Nutrition)

► Show Figures

Figure 1

18 pages, 7115 KB

Open AccessArticle

Unveiling Embryonic Development of the Threatened Neotropical Fish Prochilodus vimboides (Characiformes: Prochilodontidae)

by Renato Massaaki Honji, Amanda da Silveira Guerreiro, Bruno Cavalheiro Araújo, Danilo Caneppele, Sergio Ricardo Batlouni and Renata Guimarães Moreira

Animals 2026, 16(5), 852; https://doi.org/10.3390/ani16050852 - 9 Mar 2026

Viewed by 1252

Abstract

Understanding embryonic development is fundamental to improving captive breeding protocols and supporting conservation strategies for threatened fish species. Prochilodus vimboides is a Neotropical freshwater fish for which detailed information on early ontogeny remains scarce. This study aimed to characterize the embryonic and early [...] Read more.

Understanding embryonic development is fundamental to improving captive breeding protocols and supporting conservation strategies for threatened fish species. Prochilodus vimboides is a Neotropical freshwater fish for which detailed information on early ontogeny remains scarce. This study aimed to characterize the embryonic and early larval development of P. vimboides under captive conditions. Broodstock were hormonally induced to reproduce, and extrusion occurred between 209 and 230 degree-hours after induction at 21.49 ± 0.15 °C. Embryonic development was monitored at regular intervals after fertilization (AF) using freshly collected eggs examined under a stereomicroscope. The principal developmental stages were identified, namely zygote, cleavage, including morula and blastula, gastrula, organogenesis, and hatching. Fertilized oocytes exhibited marked hydration and formation of a large perivitelline space at 15 min AF. More than 50% of embryos reached the two-blastomere stage by 20 min AF, and cleavage continued until 2 h 14 min AF. The gastrula stage was observed at 3 h 23 min AF, blastopore closure occurred at 11 h 47 min AF, and organogenesis began at 12 h 55 min AF. Complete hatching occurred at 22 h 04 min AF, and larvae subsequently initiated yolk sac absorption without cannibalistic behavior. These findings provide a species-specific developmental framework that supports captive production and conservation efforts for P. vimboides in the Paraíba do Sul River Basin. Full article

(This article belongs to the Special Issue Fish Reproductive Biology and Embryogenesis)

► Show Figures

Figure 1

19 pages, 3878 KB

Open AccessCase Report

Raising and Releasing a Single Orphaned Harbour Seal Pup—A Case Report

by Guido Dehnhardt, Barbara Fölting and Yvonne Krüger

Animals 2026, 16(5), 851; https://doi.org/10.3390/ani16050851 - 9 Mar 2026

Cited by 1 | Viewed by 567

Abstract

Well-founded animal models of maternal separation show that early postnatal separation from the mother represents a massive stress factor for the offspring, with far-reaching negative effects on their physical and behavioural development. However, studies also show that the effects of maternal separation can [...] Read more.

Well-founded animal models of maternal separation show that early postnatal separation from the mother represents a massive stress factor for the offspring, with far-reaching negative effects on their physical and behavioural development. However, studies also show that the effects of maternal separation can be at least mitigated by appropriate physical attention. In this case report, we document the successful rehabilitation of a single orphaned male harbour seal pup by attempting to substitute natural maternal attention with intensive human attention. The pup underwent excellent physical development and displayed a close bond with its carers, characterised by intensive physical contact and suckling on a carer’s hand. However, during the final rehabilitation phase in a large outdoor enclosure, the pup increasingly reduced its affiliation to its carers. After its release, long-term observations over four years documented its hunting success, corresponding optimal physical development, and successful integration into existing wild seal groups. At the age of four, the now sub-adult seal displayed socio-sexual behaviour that is typical for this age and maintained a close bond with a wild female seal for months. The report shows that the rehabilitation of orphaned seals is possible by substituting maternal care with human care, resulting in neither a fixation on humans nor modifying species-specific behaviour. Thus, the pup’s chances of survival and potential reproduction in the wild were not jeopardised by this method of rehabilitation. Full article

(This article belongs to the Special Issue The Behaviour, Needs and Welfare of Pinnipeds in Human Care)

► Show Figures

Figure 1

13 pages, 2240 KB

Open AccessArticle

Pigs with CD163 Mutation Conferred PRRSV Resistance

by Changbao Wu, Heyao Wang, Wei Zhang, Miaomiao Cheng, Yang Wang, Lian Chen, Chao Tang, Yanfeng Dai and Liping Zhang

Animals 2026, 16(5), 850; https://doi.org/10.3390/ani16050850 - 9 Mar 2026

Cited by 1 | Viewed by 745

Abstract

Porcine reproductive and respiratory syndrome (PRRS), which is caused by the porcine reproductive and respiratory syndrome virus (PRRSV), results in substantial economic losses for the global pig farming industry. A critical step in the infection process is the binding of PRRSV to the [...] Read more.

Porcine reproductive and respiratory syndrome (PRRS), which is caused by the porcine reproductive and respiratory syndrome virus (PRRSV), results in substantial economic losses for the global pig farming industry. A critical step in the infection process is the binding of PRRSV to the CD163 receptor on the surface of porcine alveolar macrophages. This study successfully generated CD163−/− Landrace pigs using CRISPR/Cas9 gene editing technology. Following an experimental challenge with two distinct Type II PRRSV strains, the edited pigs exhibited complete resistance to infection. Virological and pathological examinations confirmed the absence of viral replication and the presence of characteristic pulmonary lesions and other organ damage in CD163−/− pigs. In contrast, wild-type control pigs exhibited high viral loads and severe pulmonary lesions, as well as damage to other organs. Our findings provide direct evidence that CD163 is an essential receptor for PRRSV infection in vivo. The CD163−/− pig model offers an effective genetic strategy for breeding pigs with an inherent resistance to PRRSV. Full article

(This article belongs to the Special Issue Animal Transgene and Gene Editing Technologies: Applications and Future Perspectives)

► Show Figures

Figure 1

16 pages, 286 KB

Open AccessArticle

Effects of Variable Dietary Sorghum Proportions on the In Vitro Digestibility of Nutrients for Ruminants

by Narimane Kellali, Iván Mateos, Cristina Saro, Nedjoua Lakhdara, Mustapha Kerrour and María José Ranilla

Animals 2026, 16(5), 849; https://doi.org/10.3390/ani16050849 - 8 Mar 2026

Cited by 1 | Viewed by 570

Abstract

This study evaluated the effects of increasing proportions of sudangrass sorghum forage in ruminant diets, with or without polyethylene glycol (PEG), on rumen fermentation, gas and methane production, nutrient digestibility, and protein fermentation metabolites. Three experimental diets containing 20%, 40%, or 60% sorghum [...] Read more.

This study evaluated the effects of increasing proportions of sudangrass sorghum forage in ruminant diets, with or without polyethylene glycol (PEG), on rumen fermentation, gas and methane production, nutrient digestibility, and protein fermentation metabolites. Three experimental diets containing 20%, 40%, or 60% sorghum forage (S20, S40, and S60) were incubated in vitro with cattle rumen fluid. Incubations were performed with or without PEG used as a tannin-binding agent. After 24 h of incubation, gas and methane production, in vitro dry matter digestibility (DMD), neutral detergent fiber digestibility (NDFD), ammonia nitrogen concentration (N-NH₃), and volatile fatty acid (VFA) production and profiles were measured. Increasing sorghum inclusion resulted in a significant reduction in DMD (p = 0.0012). In contrast, NDFD increased (p = 0.0005), likely due to differences in lignin content among diets. Methane production was unaffected by the proportion of sorghum, despite the increasing tannin content. PEG supplementation significantly increased N-NH₃ concentration (p = 0.042) and isobutyric molar proportion (p < 0.0001), indicating enhanced rumen protein degradation following tannin neutralization. The total VFA concentration was not influenced by either sorghum level or PEG treatment. However, higher sorghum inclusion was associated with shifts in the VFA profiles toward higher acetate (p = 0.0023) and lower butyrate proportions (p = 0.0114). Overall, the results suggest that moderate levels of condensed tannins (CTs) in sorghum forage may alter rumen fermentation patterns without markedly reducing methane production. PEG supplementation further confirmed the biological activity of tannins, especially regarding protein metabolism. Therefore, sudangrass sorghum may be considered a viable forage option for ruminant diets, provided its inclusion level and tannin effects are carefully managed. Full article

(This article belongs to the Section Animal Nutrition)

18 pages, 1290 KB

Open AccessArticle

Phytochemical Profiling, Antioxidant and Antibacterial Activities of Persicaria odorata Ethanolic Leaf Extract and Evaluation of Growth Performance, Disease Resistance, and Gene Expression in Labeo chrysophekadion (Bleeker, 1849)

by Sontaya Sookying, Dutrudi Panprommin, Grissana Pook-in, Pinhatai Pardubyew, Thanatip Kaeothep, Tunyatorn Nakwong and Paiboon Panase

Animals 2026, 16(5), 848; https://doi.org/10.3390/ani16050848 - 8 Mar 2026

Viewed by 592

Abstract

Persicaria odorata is a medicinal plant recognized for its diverse biological activities. This study investigated the phytochemical profile, antioxidant and antibacterial activities of P. odorata ethanolic leaf extract (POE), as well as its effects on growth performance, disease resistance, and gene expression in [...] Read more.

Persicaria odorata is a medicinal plant recognized for its diverse biological activities. This study investigated the phytochemical profile, antioxidant and antibacterial activities of P. odorata ethanolic leaf extract (POE), as well as its effects on growth performance, disease resistance, and gene expression in Labeo chrysophekadion (Bleeker, 1849) over 150 days. Qualitative screening of POE confirmed the presence of phenolics and flavonoids associated with antioxidant and antibacterial properties. Quantitative analysis determined total phenolic and flavonoid contents in POE, and HPLC confirmed the presence of quercetin and catechin. The extract exhibited notable antioxidant capacity and antibacterial activity against selected pathogens. Dietary supplementation with POE at different inclusion levels resulted in numerically higher growth indices than the control, although differences were not statistically significant (p > 0.05). Fish fed 0.50 and 0.75 g/kg POE showed reduced cumulative mortality compared with the control and 0.25 g/kg group (p < 0.05). POE supplementation also upregulated IL-1β and IGF-1 expression, suggesting enhanced immune responses and growth regulation. Overall, POE shows potential as a natural feed additive to support fish health and disease resistance in aquaculture. Full article

(This article belongs to the Section Animal Nutrition)

► Show Figures

Figure 1

16 pages, 1706 KB

Open AccessSystematic Review

Anuran amphibian Hemoparasites over the Last Century: Advances, Challenges, and Future Prospects: A Systematic Review

by Paula Andrea Yepes, Lucas S. Barrientos and Adriana Pulido-Villamarín

Animals 2026, 16(5), 847; https://doi.org/10.3390/ani16050847 - 8 Mar 2026

Viewed by 967

Abstract

Hemoparasites are intracellular organisms that affect groups of vertebrates, including amphibians, yet their diversity, distribution, and ecological effects remain partially understood. This systematic review analyzed advances over the past one hundred years in the study of hemoparasites in anurans, emphasizing: (1) the geographical [...] Read more.

Hemoparasites are intracellular organisms that affect groups of vertebrates, including amphibians, yet their diversity, distribution, and ecological effects remain partially understood. This systematic review analyzed advances over the past one hundred years in the study of hemoparasites in anurans, emphasizing: (1) the geographical distribution of studies, (2) the diagnostic techniques employed, and (3) the diversity of parasites and hosts involved. To this end, 83 scientific articles published between 1924 and 2024 were reviewed, following PRISMA guidelines and within an adapted PICO framework. The results reveal a notable increase in publications since the 2000s, with a concentration in North and South America. Morphological studies predominated (69.9%), while only 22.9% combined morphological and molecular techniques and 9.6% used only molecular techniques. Protozoan genera such as Hepatozoon and Trypanosoma, as well as nematodes (microfilariae) of the family Filaroidea, were frequently reported (69.7%); on the host side, the most studied anuran families were Ranidae (34.6%), Hylidae (6.9%), and Bufonidae (21.5%). Important methodological gaps were evident, particularly biodiverse tropical regions such as Central Africa, Southeast Asia, the Andean-Amazon, and Central America. These findings underscore the need to strengthen local research capacities, expand sampling efforts, and standardize diagnostic protocols to advance knowledge of hemoparasite diversity, ecology, and their role in ecosystem health. Full article

(This article belongs to the Special Issue Wild and Domestic Animal Hemoparasites)

► Show Figures

Figure 1

19 pages, 460 KB

Open AccessReview

Effect of Omega-3 Fatty Acid Supplementation on Broilers’ Health and Meat Quality—Systematic Review

by Peter Ayodeji Idowu, Tshilidzi Cynthia Negogogo and Takalani J. Mpofu

Animals 2026, 16(5), 846; https://doi.org/10.3390/ani16050846 - 8 Mar 2026

Cited by 1 | Viewed by 1151

Abstract

Omega-3 polyunsaturated fatty acids are increasingly applied as functional nutrients to improve broiler health, metabolic resilience, and meat quality. However, studies remain inconsistent regarding optimal inclusion levels and their efficacy. This systematic review was conducted following PRISMA 2020 guidelines. This study included 69 [...] Read more.

Omega-3 polyunsaturated fatty acids are increasingly applied as functional nutrients to improve broiler health, metabolic resilience, and meat quality. However, studies remain inconsistent regarding optimal inclusion levels and their efficacy. This systematic review was conducted following PRISMA 2020 guidelines. This study included 69 experimental studies that met predefined eligibility criteria, requiring broiler-specific trials with clear reporting of omega-3 source, inclusion level, and measurable outcomes related to growth, immune or antioxidant function, gut morphology, fatty acid composition, or meat quality. Searches were performed in Scopus and Web of Science for peer-reviewed articles published between 2020 and 2025. Across studies, supplementation with fish oil, flaxseed, or algal oil consistently increased the deposition of long-chain omega-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in broiler tissues. Omega-3 inclusion improved immune modulation, gut morphology, and antioxidant enzyme activity while reducing inflammatory markers and lipid peroxidation. Meat quality benefits included lower n-6:n-3 ratios by improving tenderness and juiciness and enhanced oxidative stability with no detrimental effects on growth performance. Nevertheless, responses varied with broiler strain, dietary formulation, feeding phase, antioxidant protection, and environmental conditions. As high levels of marine oils remained susceptible to oxidative degradation. Overall, omega-3 supplementation represents a viable and scalable precision-nutrition strategy for enriching poultry meat and supporting broiler health. Future research should prioritize cost-effective algal sources, establish phase-specific inclusion thresholds across genetic lines, and evaluate synergy with natural antioxidants and bioactive compounds to optimize oxidative stability, shelf life, and consumer acceptance. Full article

(This article belongs to the Special Issue Strategies to Improve Gut Health and Immunity in Monogastric Animals)

► Show Figures

Figure 1

20 pages, 800 KB

Open AccessArticle

Phytosynbiotic Containing Double-Layer Microencapsulated Pediococcus acidilactici V202 and Tiliacora triandra Leaf Extract Improve Growth Performance and Gut Health in Broiler Chickens

by Manatsanun Nopparatmaitree, Juan J. Loor, Chaiwat Arjin, Noraphat Hwanhlem, Pranpriya Sudchamrong, Buachompooputr Buapa, Payungsuk Intawicha and Tossaporn Incharoen

Animals 2026, 16(5), 845; https://doi.org/10.3390/ani16050845 - 7 Mar 2026

Viewed by 666

Abstract

This study investigated the effects of a novel phytosynbiotic feed supplement derived from double-layer microencapsulated Pediococcus acidilactici V202 and Tiliacora triandra leaf extract (DMP) on the growth performance, nutrient utilization, gut fermentation, intestinal morphology, and cecal microbiota of broiler chickens. A total of [...] Read more.

This study investigated the effects of a novel phytosynbiotic feed supplement derived from double-layer microencapsulated Pediococcus acidilactici V202 and Tiliacora triandra leaf extract (DMP) on the growth performance, nutrient utilization, gut fermentation, intestinal morphology, and cecal microbiota of broiler chickens. A total of 250 one-day-old male Ross 308 broilers were randomly assigned to five dietary treatments: basal control diet, antibiotic growth promoter (AGP) with chlortetracycline at 0.07%, and DMP supplementation at 0.25, 0.50, or 1.00% (w/w) for 42 days. Compared with the control diet, feeding the DMP led to linear or quadratic responses (p < 0.05) on average daily gain, feed efficiency, productive index, and economic returns. Apparent digestibility of dry matter, crude protein, and apparent metabolizable energy was enhanced in DMP-fed broilers, indicating improved nutrient utilization efficiency. These performance responses were accompanied by pronounced alterations in cecal fermentation, characterized by increased lactic acid, total volatile fatty acids, and particularly acetic and butyric acid levels (p < 0.01). Microbiome analysis revealed that the DMP selectively enriched fermentative SCFA-producing bacterial orders, including Lachnospirales, Oscillospirales, and Lactobacillales. It also reduced the relative abundance of less desirable taxa. As evidenced by an increased villus height and surface area in the duodenum and jejunum, along with a higher villus height-to-crypt depth ratio in the ileum, feeding the DMP also enhanced small intestinal morphology. These coordinated morphological adaptations are indicative of enhanced epithelial maturation and reduced crypt hyperplasia, likely mediated by elevated microbial SCFA production in the gut. In conclusion, the DMP improved broiler growth performance by coordinating the modulation of the gut microbiota, SCFA levels, and intestinal morphology, resulting in enhanced nutrient digestibility and productivity. This phytosynbiotic strategy represents a sustainable plant-based alternative to antibiotic growth promoters for environmentally responsible poultry production. Full article

(This article belongs to the Special Issue Biological Activities of Natural Compounds and Their Application in Animal Health)

► Show Figures

Figure 1

22 pages, 3376 KB

Open AccessArticle

Effects of Dietary Non-Fibrous Carbohydrate to Neutral Detergent Fiber Ratio on Apparent Digestibility, Fecal Microbiota, and Plasma Metabolomics in Yili Horses

by Mengfei Li, Zihao Xu, Long Sun, Zhiqiang Cheng, Yingying Yu, Yong Chen, Fengming Li and Changjiang Zang

Animals 2026, 16(5), 844; https://doi.org/10.3390/ani16050844 - 7 Mar 2026

Viewed by 596

Abstract

This study aimed to investigate the effects of dietary NFC/NDF ratio on nutrient apparent digestibility, fecal fermentation parameters, microbial diversity, and plasma metabolomics in Yili horses. Twenty-four healthy Yili horses with similar body weights (406 ± 22.73 kg) were divided into four groups, [...] Read more.

This study aimed to investigate the effects of dietary NFC/NDF ratio on nutrient apparent digestibility, fecal fermentation parameters, microbial diversity, and plasma metabolomics in Yili horses. Twenty-four healthy Yili horses with similar body weights (406 ± 22.73 kg) were divided into four groups, each with six replicates: the Control Group (CG), Low-NFC Group (LG), Medium-NFC Group (MG), and High-NFC Group (HG). The experiment lasted 52 d, comprising a 7-day adaptation period and a 45-day experimental period. Total fecal collection was conducted from days 41 to 45 to calculate nutrient apparent digestibility. On the final day, rectal fecal samples and blood samples were collected for full-length 16S rRNA gene sequencing and plasma metabolomics analysis. The results revealed the following findings: (1) The apparent digestibility of crude protein (CP) in the MG and HG groups was significantly higher than in the CG (p < 0.01), and significantly higher in the LG group compared to the CG (p < 0.05). (2) Significant differences were observed in fecal pH, propionate concentration, and the acetate-to-propionate ratio between the CG and the experimental groups (p < 0.05). (3) At the phylum level, Firmicutes, Bacteroidota, and Verrucomicrobiota were dominant in the fecal microbiota of all groups. PICRUSt2 prediction indicated that the MG and HG groups primarily enhanced energy conversion efficiency through amino acid metabolism and pantothenate and CoA biosynthesis metabolic pathways. (4) A total of 204 differential metabolites were identified between the CG and MG groups, with 98 upregulated and 106 downregulated in the MG group compared to the CG. These metabolites were mainly enriched in pantothenate and CoA biosynthesis, fructose and mannose metabolism, pyruvate metabolism, and starch and sucrose metabolism pathways. In summary, appropriately increasing NFC/NDF content influences the gut microbiota composition and energy metabolism of Yili horses, thereby effectively improving their digestion and absorption of dietary nutrients. Full article

(This article belongs to the Special Issue Dietary Regulation of the Rumen Microbiome and Fermentation)

► Show Figures

Figure 1

13 pages, 1357 KB

Open AccessArticle

ELAVL1 Promotes Proliferation and Inhibits Apoptosis of the Marek’s Disease Virus (MDV)-Transformed Cell Line MSB1 via the COX-2/PGE2 Pathway

by Lei He, Dong-Mei Zhan, Hui Peng, Meng-Ru Gao, Jian Chen, Yan-Yan Jia, Cheng-Shui Liao, Song-Biao Chen, Ke Ding and Zu-Hua Yu

Animals 2026, 16(5), 843; https://doi.org/10.3390/ani16050843 - 7 Mar 2026

Viewed by 593

Abstract

Marek’s disease (MD), caused by the oncogenic Marek’s disease virus (MDV), is a highly contagious avian infection that induces lymphoproliferative tumors. The RNA-binding protein ELAVL1 is known to regulate tumor cell proliferation and apoptosis, but its role in MDV-induced oncogenesis remains unclear. This [...] Read more.

Marek’s disease (MD), caused by the oncogenic Marek’s disease virus (MDV), is a highly contagious avian infection that induces lymphoproliferative tumors. The RNA-binding protein ELAVL1 is known to regulate tumor cell proliferation and apoptosis, but its role in MDV-induced oncogenesis remains unclear. This study investigated whether ELAVL1 modulates proliferation and apoptosis in the MDV-transformed MSB1 cell line and whether its effects involve the cyclooxygenase-2 (COX-2)/prostaglandin E2 (PGE2) pathway. MSB1 cells were transiently transfected with ELAVL1-overexpressing plasmids (pEGFP-C-ELAVL1) or ELAVL1-specific siRNA, with expression confirmed by real-time PCR (qRT-PCR). Cell proliferation was assessed using the CCK-8 assay, while cell cycle distribution and apoptosis rates were analyzed by flow cytometry. COX-2 and PGE2 expression levels were determined by qRT-PCR, Western blotting, and ELISA. Overexpression of ELAVL1 significantly promoted the proliferation of MSB1 cells, decreased transition into the G1 phase, increased the proportions of S and G2 phase cells, and suppressed apoptosis. Correspondingly, both mRNA and protein levels of COX-2 and PGE2 were significantly elevated. Conversely, ELAVL1 knockdown significantly inhibited proliferation, induced G1 phase arrest, decreased S phase cells, and significantly decreased COX-2 and PGE2 expression. These findings indicate that ELAVL1 promotes proliferation and inhibits apoptosis in MDV-transformed MSB1 cells, potentially via the COX-2/PGE2 signaling pathway. Full article

(This article belongs to the Special Issue Pathogenesis, Immunology and Epidemiology of Veterinary Viruses—2nd Edition)

► Show Figures

Figure 1

27 pages, 3406 KB

Open AccessArticle

Animal Welfare Monitor: Raising the Bar for Species-Specific Welfare Evaluation Using Welfare Quality^® Principles

by Amélie Romain, Léa Briard, Gwenaël Leroutier, Marine Parker, Baptiste Chenet, Constance Wagner, Alexandre Petry and Benoît Quintard

Animals 2026, 16(5), 842; https://doi.org/10.3390/ani16050842 - 7 Mar 2026

Viewed by 879

Abstract

Assessing zoo animal welfare can involve generic evaluations or targeted, species-specific protocols. While the latter offer greater precision, their development is often limited by species diversity and the lack of validated indicators. The Animal Welfare Monitor^® (AWM) protocol addresses these challenges by [...] Read more.

Assessing zoo animal welfare can involve generic evaluations or targeted, species-specific protocols. While the latter offer greater precision, their development is often limited by species diversity and the lack of validated indicators. The Animal Welfare Monitor^® (AWM) protocol addresses these challenges by adapting Welfare Quality^® principles to zoological contexts. Its core innovation is a four-level hierarchical structure (base, order, family, species) linking broad taxonomic knowledge to species-level protocols. This enables tailored questionnaires for each species, including data-deficient taxa, by leveraging information from related groups. Questionnaires, covering housing, nutrition, health, and behaviour, are complemented by behavioural observations. AWM currently covers 87 species (69 mammals, 15 birds, 2 amphibians, 1 reptile) and constitutes a substantial database of species-specific welfare assessment protocols embedded within a single, standardised methodological framework. Between 2021 and 2025, 14 zoos conducted over 1000 assessments and 15,000 behavioural observations, demonstrating the protocol’s feasibility in routine operations. AWM integrates data entry with visual documentation, such as photographs of enclosures or enrichment, which add context, enhance decision-making, and strengthen long-term records. While refinements such as group-level assessment remain, AWM offers a scalable, flexible tool combining scientific rigour with operational applicability, supporting positive welfare outcomes across diverse zoological institutions. Full article

(This article belongs to the Special Issue Best Practices for Zoo Animal Welfare Management)

► Show Figures

Figure 1

14 pages, 15162 KB

Open AccessArticle

Periostracum Formation in Sepia officinalis and Loligo vulgaris and Homology with Other Molluscs

by Ernesto Ruiz-Villaespesa, Antonio G. Checa, Cristina Lucena-Serrano and Carmen Salas

Animals 2026, 16(5), 841; https://doi.org/10.3390/ani16050841 - 7 Mar 2026

Viewed by 485

Abstract

The periostracum is the outermost shell layer and the first produced during shell formation in molluscs. This organic layer isolates the extrapallial space from the external environment and provides a scaffold for subsequent calcification. In cephalopods with an internal shell, some organic shell [...] Read more.

The periostracum is the outermost shell layer and the first produced during shell formation in molluscs. This organic layer isolates the extrapallial space from the external environment and provides a scaffold for subsequent calcification. In cephalopods with an internal shell, some organic shell structures are putatively homologous to the periostracum of other molluscan groups. However, neither their detailed structure nor their mode of formation has been described, leaving the extent of this homology unresolved. To address this issue, we investigated the morphology and formation of the organic layer of the dorsal shield and the gladius in embryos of the cuttlefish Sepia officinalis Linnaeus, 1758, and the squid Loligo vulgaris Lamarck, 1798, respectively, using light microscopy and transmission electron microscopy. In both species, the periostracum forms within a periostracal groove located along the lateral and anterior margins of the shell sac. As in other molluscs, secretions from columnar cells at the bottom of the groove produce a dense layer, while a translucent layer is subsequently added beneath it through secretions from cuboidal cells. The main difference is the absence of both a pellicle and of the specialized glandular cells that typically secrete it at the bottom of the periostracal groove. Full article

(This article belongs to the Special Issue Recent Advances in Cephalopod Biology Research)

► Show Figures

Figure 1

19 pages, 7656 KB

Open AccessArticle

Gut Microbiome Signatures Across Migratory, Sedentary, and Aquaculture Ecotypes of Coilia nasus

by Xue Liu, Congping Ying, Fengjiao Ma, Yanping Yang and Kai Liu

Animals 2026, 16(5), 840; https://doi.org/10.3390/ani16050840 - 7 Mar 2026

Cited by 1 | Viewed by 581

Abstract

Coilia nasus, a typical species with migratory–sedentary polymorphism, shows different intestinal microbiota characteristics among its different ecotypes. This is attributed to differences in feeding habits and habitat environments (such as water temperature, salinity, etc.). This study constructed a database of intestinal microbiota [...] Read more.

Coilia nasus, a typical species with migratory–sedentary polymorphism, shows different intestinal microbiota characteristics among its different ecotypes. This is attributed to differences in feeding habits and habitat environments (such as water temperature, salinity, etc.). This study constructed a database of intestinal microbiota for three ecological types of C. nasus, namely migratory type (comprising marine populations and freshwater populations), sedentary type and aquaculture-reared type, through 16S rRNA amplicon sequencing technology. This study investigates the ecological mechanisms underlying microbiota differentiation, focusing on three key drivers: environmental selection, host nutritional metabolism requirements, and host life history strategies. The results showed that the core flora of C. nasus consisted of Firmicutes, Proteobacteria, and Actinobacteria. Both the depletion of microbial taxa and the enrichment of marine-adapted bacterial lineages—including Proteobacteria and Psychrobacter—are associated with elevated salinity in the migratory marine population of C. nasus. In contrast, the elevated relative abundance of Actinobacteria in aquaculture-reared C. nasus is likely attributable to dietary supplementation with protein- and lipid-rich artificial feed. Functional correlation analysis holds promise for partially predicting the microbiota’s metabolic functional succession patterns. The dominance of Pseudomonas_E in the migratory freshwater population is consistent with its well-documented physiological versatility and adaptive capacity in dynamically fluctuating aquatic habitats. The elevated abundance of Cyanobacteria in the sedentary population C. nasus coincides with the water bloom in their habitat, suggesting that the structure of the microbiota may serve as a novel biomarker for indicating the ecosystem. In conclusion, this study identifies potential molecular markers for tracing genetic resources and distinguishing ecological types of C. nasus, while establishing a theoretical foundation for elucidating the co-evolutionary dynamics between fish hosts and their associated microbiota—and thereby informing both conservation strategies for wild populations and microbiota-informed aquaculture practices. Full article

(This article belongs to the Special Issue Gut Microbiota in Aquatic Animals)

► Show Figures

Figure 1

11 pages, 4147 KB

Open AccessCase Report

Treatment of Feline Lung–Digit Syndrome with Toceranib Phosphate: Prolonged Survival and Novel Metastatic Findings

by Inês Cabral, Gustavo Matos, Gabriela Fernandes Silva, Fátima Carvalho and Irina Amorim

Animals 2026, 16(5), 839; https://doi.org/10.3390/ani16050839 - 7 Mar 2026

Viewed by 1426

Abstract

Feline pulmonary carcinomas are rare and often carry a poor prognosis, particularly when associated with feline lung–digit syndrome. We report a cat with primary pulmonary carcinoma and extensive metastases—including digits, pleura, mandible, scapula, spleen, skeletal muscle, and distant lymph nodes—supporting the broader “MODAL [...] Read more.

Feline pulmonary carcinomas are rare and often carry a poor prognosis, particularly when associated with feline lung–digit syndrome. We report a cat with primary pulmonary carcinoma and extensive metastases—including digits, pleura, mandible, scapula, spleen, skeletal muscle, and distant lymph nodes—supporting the broader “MODAL syndrome” concept. Palliative therapy with toceranib phosphate and meloxicam achieved prolonged survival and excellent quality of life, with no adverse effects despite dose escalation. Rapid progression after discontinuation suggests a role for toceranib in delaying tumour growth. Immunohistochemistry revealed c-kit expression in one metastatic lesion but not in the primary tumour or most metastases, highlighting intratumoral heterogeneity and the complexity of targeted therapy. The observed benefit likely reflects toceranib’s multi-target activity (VEGFR2, PDGFR), impacting angiogenesis and tumour progression. This case represents the first report of toceranib phosphate use in feline pulmonary carcinoma and underscores its potential as a palliative option. Full article

(This article belongs to the Special Issue Comparative Oncology in Animals: Advancements in Diagnostics and Treatment)

► Show Figures

Figure 1

30 pages, 410 KB

Open AccessReview

The Influence of Environmental Conditions and Husbandry Practices on Goat Welfare

by Renata Pilarczyk, Małgorzata Bąkowska, Agnieszka Tomza-Marciniak, Jan Udała, Beata Seremak, Ewa Kwita, Piotr Sablik and Bogumiła Pilarczyk

Animals 2026, 16(5), 838; https://doi.org/10.3390/ani16050838 - 7 Mar 2026

Viewed by 643

Abstract

Goat (Capra hircus) welfare is an important issue in any farming system. The aim of the study was a comprehensive analysis of the impact of environmental factors and farming practices on the welfare of goats, with particular attention to physical, behavioural, [...] Read more.

Goat (Capra hircus) welfare is an important issue in any farming system. The aim of the study was a comprehensive analysis of the impact of environmental factors and farming practices on the welfare of goats, with particular attention to physical, behavioural, and emotional aspects. It includes a review of the up-to-date literature on the effects of environmental conditions including air temperature, air humidity, space, feeding systems, social relationships (mother–offspring, human–animal, animal–animal), zootechnical procedures (dehorning, castration, hoof trimming) and welfare assessment methods. It compares the AWIN, Anzuino, Muri and Leite protocols for assessing goat welfare and their application in the Five Domain Model. Goat welfare is strongly influenced by their environment, nutrition and socialisation: heat stress and confined space cause physiological disorders, decreased immunity and increased aggressive behaviour and a monotonous diet leads to frustration and reduced cognitive activity, whereas positive early contact with humans reduces anxiety and maintaining the mother–kid bond supports the social development of young goats. Furthermore, significant improvements in welfare and stress reduction can be achieved by providing anaesthesia and painkillers where necessary to minimise pain and enriching the environment with items that support natural behaviour, such as platforms, brushes and items for cognitive tasks. In general, the keeper should take a holistic approach, combining environmental optimisation, humane husbandry practices and regular monitoring using validated assessment protocols to improve welfare. These measures are both an ethical obligation and a prerequisite for animal health and production efficiency. Nevertheless, there is a need for further research focussing on the development of non-invasive assessment methods and innovative forms of environmental enrichment. Full article

(This article belongs to the Section Animal Welfare)

14 pages, 758 KB

Open AccessArticle

Tree Composition, Niche Characteristics, and Mammal Habitat Use Across Different Types of Forests in Wanglang Nature Reserve

by Chenhui Qu, Chenggong Song, Dongwei Kang and Yanhong Liu

Animals 2026, 16(5), 837; https://doi.org/10.3390/ani16050837 - 7 Mar 2026

Viewed by 441

Abstract

Effectiveness of forest restoration efforts depends on the methods employed. Here, we compared tree species composition, niche characteristics, and mammal habitat use in primary, secondary, and artificial forests in Wanglang Nature Reserve. Results showed that primary forests were mainly indicated by Abies fargesii [...] Read more.

Effectiveness of forest restoration efforts depends on the methods employed. Here, we compared tree species composition, niche characteristics, and mammal habitat use in primary, secondary, and artificial forests in Wanglang Nature Reserve. Results showed that primary forests were mainly indicated by Abies fargesii var. faxoniana (Af), Picea purpurea (Pp), and Juniperus saltuaria (Js); secondary forests were mainly indicated by Af and Betula albosinensis (Ba); and artificial forests were mainly indicated by Picea asperata (Pa) and Acer caesium (Ac). Af had the broadest niche breadth in natural forests, and Pa had the broadest niche breadth in artificial forests. Low niche overlap among common species was observed in natural forests, whereas high niche overlap between Pa and Ba occurred in artificial forests. Interspecific correlations showed that Af was negatively correlated with Pp in primary forests and Populus szechuanica (Ps) in secondary forests. In artificial forests, Af and Ac were positively correlated. Furthermore, no traces of the three National Class I protected species were found in artificial forests, while traces of two representative mammals were associated with Af. These findings highlight the differences among the three types of forests. Full article

(This article belongs to the Special Issue Recent Advances in Giant Panda and Umbrella Species Conservation: Management and Strategies)

► Show Figures

Figure 1

14 pages, 3279 KB

Open AccessArticle

Functional Characterization of IGF2BP1, CDC25A, and RXFP2 Genes: Implications for Ovarian Function and Reproductive Regulation in Goats

by Haiyan Yang, Qiancheng Ma, Zhiying Wang, Shan Zhang, Luqi Wang, Haijing Zhu, Xianyong Lan, Ke Wang and Chuanying Pan

Animals 2026, 16(5), 836; https://doi.org/10.3390/ani16050836 - 7 Mar 2026

Viewed by 427

Abstract

Goat reproductive performance is a key determinant of the productivity and economic value of goat farming, especially in meat and milk production. In a previous study, to investigate the genetic basis of prolificacy, we divided goats into groups according to their consistent reproductive [...] Read more.

Goat reproductive performance is a key determinant of the productivity and economic value of goat farming, especially in meat and milk production. In a previous study, to investigate the genetic basis of prolificacy, we divided goats into groups according to their consistent reproductive performance (producing either single kids or twins) over five consecutive kidding cycles, and performed whole-genome resequencing and RNA-seq analysis on their ovarian tissues. Through integrated analysis, we identified three candidate genes—IGF2BP1 (insulin-like growth factor 2 mRNA-binding protein 1), CDC25A (cell division cycle 25A), and RXFP2 (relaxin family peptide receptor 2)—as potential key regulators of reproductive capacity. Using goat ovarian granulosa cells, we systematically assessed the impact of each gene through gain- and loss-of-function experiments. Overexpression of IGF2BP1 promoted cell proliferation and suppressed apoptosis, underscoring its role in maintaining cellular viability. Conversely, its knockdown significantly impeded growth and induced cell death. Similarly, CDC25A enhanced granulosa cell proliferation, whereas its knockdown led to marked growth impairment and increased apoptosis. Proliferation was also enhanced by RXFP2 overexpression but impaired upon its knockdown, suggesting that RXFP2 is functionally important for follicular development. Collectively, these findings establish IGF2BP1, CDC25A, and RXFP2 as fundamental regulators of granulosa cell dynamics and ovarian follicular development, providing crucial functional insights and promising targets for genetic selection to enhance reproductive efficiency in goats. Full article

(This article belongs to the Special Issue Genetics and Breeding for Enhancing Production Traits in Ruminants)

► Show Figures

Figure 1

16 pages, 2460 KB

Open AccessArticle

Tactile Preferences in Goats: Implications for Animal-Assisted Interventions

by Patrycja Magdalena Masier, Agnieszka Ziemiańska, Kamila Janicka, Wiktoria Janicka, Marta Wójcik and Iwona Rozempolska-Rucińska

Animals 2026, 16(5), 835; https://doi.org/10.3390/ani16050835 - 7 Mar 2026

Viewed by 544

Abstract

Tactile contact is a key element of animal-assisted interventions (AAI), yet its effects on the animals involved remain poorly understood. Seven adult goats were included in the experiment, which examined three factors influencing tactile contact: the body region (head/neck, trunk, hindquarters; R1–R3), the [...] Read more.

Tactile contact is a key element of animal-assisted interventions (AAI), yet its effects on the animals involved remain poorly understood. Seven adult goats were included in the experiment, which examined three factors influencing tactile contact: the body region (head/neck, trunk, hindquarters; R1–R3), the ordinal position in the stroking sequence (first, second, third; O1–O3) and the location context (slightly enriched goat house and a highly enriched pasture; L1, L2). It was found that when stroking occurred first in the position of the sequence (O1), the duration of touch was significantly shorter (p < 0.0001). In addition, it was observed that when the session started with the trunk (R2), the total contact duration (T) was significantly longer (p < 0.039). No differences were found between the duration of contact in different locations (p = 0.179). These results may indicate that the initial phase was critical for the overall contact. Furthermore, the longer duration of contact, when initiated on the trunk, may indicate a relative preference for this body region. These findings suggest that human contact remains equally attractive to goats regardless of the enrichment of their location. Full article

(This article belongs to the Section Animal Welfare)

► Show Figures

Figure 1

9 pages, 436 KB

Open AccessArticle

Assessment of Compliance with Animal Welfare Requirements Across Poultry Species and Production Categories

by Eva Justova, Vladimir Vecerek, Zbynek Semerad, Marijana Vucinic and Eva Voslarova

Animals 2026, 16(5), 834; https://doi.org/10.3390/ani16050834 - 7 Mar 2026

Viewed by 532

Abstract

Animal welfare is a key component of sustainable poultry production and is routinely monitored through official veterinary inspections. The aim of this study was to determine the level of welfare compliance among different poultry species and production categories, to compare compliance levels across [...] Read more.

Animal welfare is a key component of sustainable poultry production and is routinely monitored through official veterinary inspections. The aim of this study was to determine the level of welfare compliance among different poultry species and production categories, to compare compliance levels across these groups, and to assess long-term trends using official inspection data. The study was based on the results of supervisory inspections conducted by veterinary inspectors in poultry farms in the Czech Republic between 2016 and 2024. Welfare compliance was evaluated in laying hens, broiler chickens, turkeys, ducks, and geese using a standardized system of welfare assessment checkpoints applied during official controls. Inspections were classified as compliant or non-compliant based on the presence or absence of deficiencies, and overall compliance levels were calculated as the proportion of animals kept in farms with compliant inspections. Across the entire study period, the proportion of poultry kept in farms with compliant inspections ranged from 82.8% to 98.4%, with the highest compliance level observed in turkeys, followed by ducks and broiler chickens, while the lowest compliance level was recorded in geese. Differences among poultry species and categories were statistically significant (p < 0.05). Comparison of two time periods (2016–2018 and 2022–2024) revealed significant improvements in compliance for broiler chickens, ducks, and geese, whereas significant declines were observed for laying hens and turkeys (p < 0.001). These results demonstrate clear differences in welfare compliance among poultry species and categories and indicate that compliance trends over time are not uniform across the poultry sector. Official veterinary inspection data provide a valuable tool for large-scale assessment of welfare compliance and for identifying poultry sectors that may benefit from targeted welfare improvement measures. Full article

(This article belongs to the Section Poultry)

► Show Figures

Figure 1

16 pages, 1484 KB

Open AccessArticle

Primary Culture and Characterization of a Crucian Carp (Carassius carassius) Osteoblast Cell Line (COBC) and the Effects of Hypoxia on Its Differentiation

by Zaozao Guo, Jiamin Liu, Songlin Chen, Guodong Zheng and Shuming Zou

Animals 2026, 16(5), 833; https://doi.org/10.3390/ani16050833 - 6 Mar 2026

Viewed by 525

Abstract

In the present study, vertebral bone tissues derived from Chongming crucian carp (Carassius carassius), a dominant species during the summer and autumn seasons on Chongming Island in the lower Yangtze River, were used to establish and characterize a Carassius carassius osteoblast [...] Read more.

In the present study, vertebral bone tissues derived from Chongming crucian carp (Carassius carassius), a dominant species during the summer and autumn seasons on Chongming Island in the lower Yangtze River, were used to establish and characterize a Carassius carassius osteoblast cell line (COBC). The established COBCs were assessed using chromosome analysis, osteocalcin enzyme-linked immunosorbent assay (ELISA), and osteogenesis-related gene expression analysis. Additionally, cellular responses to environmental stress were assessed. The results showed that COBC exhibited optimal proliferation in L-15 medium supplemented with 20% fetal bovine serum at 28 °C. The histochemical staining assay results were all positive, thereby confirming that the isolated cells display typical osteoblast characteristics. Quantitative PCR analysis revealed that osteogenic marker genes, including runx2a and runx2b, were expressed at significantly higher levels in COBCs than in fish tissues. Under hypoxic stress, COBCs exhibited morphological changes, an increase in cell death, significant alterations in gene expression, and variations in antioxidant enzyme activity. These responses facilitate adaptation to hypoxic stress. This study established the first osteoblast cell line of the Chongming crucian carp and characterized its biological properties and response to hypoxic stress. These findings offer a valuable in vitro cell model and technical support for research on fish bone tissue biology and the assessment of environmental stress effects. Full article

(This article belongs to the Special Issue Genetic Selection and Breeding for Improved Traits in Aquaculture Animals)

► Show Figures

Figure 1

22 pages, 5347 KB

Open AccessArticle

Boron Triggers Hepatic Ferroptosis: Unveiling the Dual-Pathogenic Nexus of Oxidative Stress and SLC7A11/GPX4 Dysregulation

by Ting He, Yumeng Li, Jiangli Huang, Weiqian Su, Siying Liu, Jinwen Quan, Gaolong Zhong, Zhonghua Liu, Dayou Shi and Wenlan Yu

Animals 2026, 16(5), 832; https://doi.org/10.3390/ani16050832 - 6 Mar 2026

Viewed by 724

Abstract

Boron compounds, classified as prohibited food additives due to their high toxicity, persist in pesticides and fertilisers, industrial processes, food supply chains, and consumer goods, perpetuating multisource exposure risks. Chronic ingestion may induce fatal hepatorenal injury; however, mechanistic insights and epidemiological surveillance remain [...] Read more.

Boron compounds, classified as prohibited food additives due to their high toxicity, persist in pesticides and fertilisers, industrial processes, food supply chains, and consumer goods, perpetuating multisource exposure risks. Chronic ingestion may induce fatal hepatorenal injury; however, mechanistic insights and epidemiological surveillance remain critically lacking amidst sector-wide regulatory gaps. This study employed integrated cellular and organismal models to elucidate the relationship between boron-induced hepatotoxicity and ferroptosis. We demonstrate that dietary boron accumulation in chicken livers is associated with histopathological damage, mitochondrial cristae dissolution and atrophy (a hallmark of ferroptosis), and elevated serum biomarkers AST and ALT. Boron exacerbates oxidative damage in hepatocytes by elevating malondialdehyde (MDA) production while modulating the Nrf2/ARE antioxidant signaling pathway—specifically downregulating key genes (Nrf2, HO-1, GCLM, CAT). Concurrently, it inhibits critical antioxidant enzymes (SOD, T-AOC), thereby depleting cellular antioxidant defenses. Crucially, boron disrupts iron homeostasis and induces ferroptosis by dysregulating the SLC7A11-GPX4 pathway: upregulating pro-ferroptotic genes (ACSL4, TF, TFR) and downregulating cytoprotective genes (SLC7A11, GPX4, FTH1). Co-treatment with the ferroptosis inhibitor ferrostatin-1 (Fer-1) attenuated boron-induced oxidative damage, whereas the ferroptosis inducer Erastin potentiated toxicity. Collectively, we pioneer the dual-pathogenic mechanism of boron hepatotoxicity—oxidative stress and ferroptotic cell death—establishing the SLC7A11/GPX4 axis as a novel therapeutic target against boron toxicity. Full article

(This article belongs to the Section Poultry)

► Show Figures

Graphical abstract

19 pages, 6543 KB

Open AccessArticle

Intratumoral Staphylococcus pseudintermedius Promotes Proliferation and Migration of CMT-U27 Cells Through the TLR2/PI3K/Akt Signaling Pathway

by Luting Luo, Jin Li and Meng Li

Animals 2026, 16(5), 831; https://doi.org/10.3390/ani16050831 - 6 Mar 2026

Viewed by 474

Abstract

Increasing evidence suggests that intratumoral microorganisms and their metabolites can modulate cancer initiation and progression. However, the composition and functional role of intratumoral bacteria in canine mammary tumors (CMTs) remain unclear. In this study, we investigated the functional significance of tumor-derived Staphylococcus in [...] Read more.

Increasing evidence suggests that intratumoral microorganisms and their metabolites can modulate cancer initiation and progression. However, the composition and functional role of intratumoral bacteria in canine mammary tumors (CMTs) remain unclear. In this study, we investigated the functional significance of tumor-derived Staphylococcus in CMTs, focusing on its effects on the proliferation and migration of CMT-U27 cells. 16S rRNA sequencing revealed reduced alpha diversity in CMT tissues, with Staphylococcus pseudintermedius identified as the most frequently isolated species. Functional assays, including CCK-8, wound healing, RT-qPCR, and Western blot analyses, demonstrated that intratumoral Staphylococcus pseudintermedius significantly enhanced cellular proliferation and migration. Mechanistically, Staphylococcus pseudintermedius significantly upregulated the expression of TLR2, as well as the phosphorylation levels of PI3K, Akt and P70S6K. The inhibition of TLR2 using C29 suppressed the mRNA expression of VEGF, MMP9, MMP2, and EGFR. Collectively, these findings indicate that intratumoral Staphylococcus pseudintermedius promotes the proliferation and migration of CMT-U27 cells through activation of the TLR2/PI3K/Akt pathway, highlighting a functional link between tumor-associated bacteria and cancer progression. Full article

(This article belongs to the Section Veterinary Clinical Studies)

► Show Figures

Figure 1

17 pages, 4288 KB

Open AccessArticle

Comparative Safety and Efficacy of Commercial Porcine Vaccines Against Mycoplasma hyopneumoniae and Porcine Circovirus Type 2 (PCV2)

by Meritxell Simon-Grifé, Alexandra Moros, Cecilia Pedernera, Ester Puigvert, Lucía Acal, Elena Plantalech, Mercè Roca, Jordi Montané, Ricard March and Marta Sitjà

Animals 2026, 16(5), 830; https://doi.org/10.3390/ani16050830 - 6 Mar 2026

Viewed by 679

Abstract

Mycoplasma hyopneumoniae (Mhyo) and porcine circovirus type 2 (PCV2) infections cause enormous economic losses in the swine production sector. Vaccination remains the most effective strategy against the complex clinical entities caused by these pathogens. While effective vaccines against PCV2 are available, immunisation against [...] Read more.

Mycoplasma hyopneumoniae (Mhyo) and porcine circovirus type 2 (PCV2) infections cause enormous economic losses in the swine production sector. Vaccination remains the most effective strategy against the complex clinical entities caused by these pathogens. While effective vaccines against PCV2 are available, immunisation against Mhyo continues to be challenging. In parallel, intradermal (ID) vaccination can optimise time and costs while providing faster and user-safe administration. This study compares the safety and efficacy of commercial porcine vaccines against Mhyo and PCV2, including intramuscular (IM) and ID ones: (1) ID recombinant Mhyo-PCV2, (2) ID recombinant PCV2 + ID inactivated Mhyo, (3) IM bivalent inactivated Mhyo and recombinant PCV2, (4) IM bivalent inactivated Mhyo and inactivated chimeric PCV1-PCV2 and (5) IM inactivated Mhyo + IM inactivated PCV2. Safety, evaluated through clinical signs, adverse effects, local reactions, and average daily weight gain, has been confirmed for all vaccines. Following challenge with both pathogens, all products reduced PCV2 viremia and faecal shedding, proving efficacy against PCV2 infection. In contrast, only the ID recombinant Mhyo-PCV2 and the IM inactivated Mhyo + IM inactivated PCV2 vaccines, significantly reduced lung lesions related to Mhyo infection, indicating a better protection compared to the other vaccines. Full article

(This article belongs to the Section Pigs)

► Show Figures

Figure 1

20 pages, 1324 KB

Open AccessReview

Clinical and Epidemiological Features of Calicivirus Infections in Cattle

by Krisztián Bányai, Valantine Ngum Ndze, Ágnes Bogdán, Attila Kiss, Tamás Tóth, Zsófia Lanszki, Gianvito Lanave, Francesco Pellegrini, Barbara Di Martino and Vito Martella

Animals 2026, 16(5), 829; https://doi.org/10.3390/ani16050829 - 6 Mar 2026

Viewed by 620

Abstract

The family Caliciviridae encompasses a diverse group of non-enveloped, positive-sense RNA viruses that are significant pathogens in veterinary medicine. This narrative review summarizes the current state of knowledge regarding the clinical, molecular, and epidemiological features of the three calicivirus genera identified in bovine [...] Read more.

The family Caliciviridae encompasses a diverse group of non-enveloped, positive-sense RNA viruses that are significant pathogens in veterinary medicine. This narrative review summarizes the current state of knowledge regarding the clinical, molecular, and epidemiological features of the three calicivirus genera identified in bovine hosts: Norovirus, Nebovirus, and Vesivirus. Bovine noroviruses and neboviruses are neglected enteric pathogens, frequently detected in association with neonatal calf diarrhea and often present in co-infections with other enteric agents. Clinical presentations for these enteric viruses range from severe, watery diarrhea to asymptomatic shedding, with distinct pathogenic profiles observed between norovirus genotypes GIII.1 and GIII.2. In contrast, the genus Vesivirus exhibits a broad host range, and bovine vesivirus strains are phylogenetically linked to vesiviruses identified in pigs and marine animals. Bovine vesivirus infections are associated with a broader spectrum of clinical manifestations, including respiratory disease, vesicular lesions, and abortion. Serological and virological surveys indicate that exposure to these viruses is ubiquitous in cattle populations globally. While direct evidence of human infection by bovine noroviruses and neboviruses remains limited, vesiviruses possess a confirmed capacity for cross-species transmission to humans. Significant knowledge gaps remain, particularly regarding in vitro culture systems, necessitating further research to facilitate vaccine development and clarify transmission dynamics. Full article

(This article belongs to the Section Cattle)

► Show Figures

Figure 1

15 pages, 1216 KB

Open AccessArticle

Olfactory Cues in the Odour Plume of Predatory Fish Reduce Foraging and Elicit Anti-Predator Behaviour in the European Green Crab Carcinus maenas

by Jonathan W. Burnett, Hannah Ohnstad, Amber M. Jones, Jörg D. Hardege and Helga D. Bartels-Hardege

Animals 2026, 16(5), 828; https://doi.org/10.3390/ani16050828 - 6 Mar 2026

Viewed by 917

Abstract

The European green crab (Carcinus maenas) is an intertidal crustacean that has extended its invasive range globally and is regarded as a major pest species for bivalves, impacting coastal food chains and aquacultures. Crabs primarily use chemosensory cues to sense their [...] Read more.

The European green crab (Carcinus maenas) is an intertidal crustacean that has extended its invasive range globally and is regarded as a major pest species for bivalves, impacting coastal food chains and aquacultures. Crabs primarily use chemosensory cues to sense their environment for feeding to avoid predation and to locate mates and induce mating. Consequently, known attractants including food baits are frequently employed in trapping and control efforts. Here, we investigate the effects of introducing a predatory fish odour to the environment and show that it elicits anti-predator behaviour in C. maenas. A 45% reduction in crab foraging and feeding behaviour was observed when predator odour was introduced compared with food control experiments. A further 23% reduction (68% reduction overall) in feeding behaviour was observed after crabs were exposed to (though housed separately from) the same seawater as the live predator for several days. This highlights the increased awareness or continuous stress of these habituated crabs towards predator odour. This study also presents potential for application as a deterrent in integrated pest management strategies for this global invader. Full article

(This article belongs to the Section Aquatic Animals)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Animals, Volume 16, Issue 5 (March-1 2026) – 164 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI