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Volume 16
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Animals, Volume 16, Issue 3 (February-1 2026) – 172 articles

Cover Story (view full-size image): Porcine species are widely used as biomedical models and represent important genetic resources for conservation, emphasizing the need for reliable in vitro follicle culture systems. Conventional two-dimensional methods often fail to maintain follicular architecture and coordinated development. Three-dimensional alginate-based systems provide structural support, though optimal conditions for porcine preantral follicles remain unclear. This study evaluated different alginate concentrations and individual versus group culture. Group-cultured follicles encapsulated in 0.5% alginate showed improved maintenance of 3D architecture, enhanced growth, and increased steroid hormone production, demonstrating an optimal balance between structural support and follicular function. View this paper
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20 pages, 4001 KB  
Article
GWAS-Identified SNPs and Candidate Genes Influencing Sex in Loach (Misgurnus anguillicaudatus)
by Junxiao Su, Han Zheng, Yumei Xiang, Yu Zeng, Can Chen, Xiaoyun Zhou and Kaijian Wei
Animals 2026, 16(3), 524; https://doi.org/10.3390/ani16030524 - 6 Feb 2026
Viewed by 967
Abstract
Loach (Misgurnus anguillicaudatus) is a small benthic fish favored by consumers in East Asia. Female loaches exhibit greater production value due to their larger size and higher meat yield. Therefore, controlling the sex of loach should be studied to implement an [...] Read more.
Loach (Misgurnus anguillicaudatus) is a small benthic fish favored by consumers in East Asia. Female loaches exhibit greater production value due to their larger size and higher meat yield. Therefore, controlling the sex of loach should be studied to implement an all-female breeding strategy. This study identified sex-related SNP markers and candidate genes in loach through GWAS. Genotyping male and female populations with WGRS revealed 84 SNPs labeled as associated with sex. Distinct SNP peaks appeared on Chr6 and Chr3, with over half of the sex-associated SNPs located on Chr6. Within these sex-associated regions, 15 key candidate genes related to sex and reproduction were identified, including hemgn and foxl2a. Among these genes, pik3cb on Chr3 and hhip, cntln, and pard3ba on Chr6 have multiple sex-associated haplotypes that can be utilized for subsequent monosex breeding. Notably, a highly male-linked SNP marker was identified on pard3ba, whose genotype is consistent with the male heterogamete (XX/XY). Through gene enrichment analysis and expression validation, signaling pathways such as TGF-β, FoxO, and mTOR were identified as being involved in sex regulation. This study provides molecular markers for sex-controlled breeding in loach, facilitating in-depth research into the mechanisms of sex regulation in loach. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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23 pages, 629 KB  
Article
The Dog–Guardian Relationship and Its Meanings: Perceptions, Expectations, and Impacts on Guardians’ Lives
by Tatiane Aparecida de Castro, Carlos Alberto Pegolo da Gama, Denise Alves Guimarães, Igor Tadeu Assis, Marco Aurélio Pereira Horta, Paulo Henrique Araújo Soares and Vinícius Silva Belo
Animals 2026, 16(3), 523; https://doi.org/10.3390/ani16030523 - 6 Feb 2026
Viewed by 1219
Abstract
Understanding the relationship between dogs and their guardians provides insight into the role that animals occupy within households and families and, by extension, within society. This perspective offers essential elements for informing policies and practices that affect outcomes in Public Health and animal [...] Read more.
Understanding the relationship between dogs and their guardians provides insight into the role that animals occupy within households and families and, by extension, within society. This perspective offers essential elements for informing policies and practices that affect outcomes in Public Health and animal welfare. This study aimed to comprehensively examine guardians’ perceptions, including expectations, meanings, and social roles attributed to dogs, in a municipality in Brazil. We conducted a qualitative and descriptive investigation grounded in Social Constructionism and Laurence Bardin’s Content Analysis, based on 40 semi-structured interviews. The findings indicate that guardians frequently describe dogs as “family members,” “sources of joy,” “companions,” or even a form of “therapy,” underscoring strong emotional bonds and noteworthy psychosocial benefits. At the same time, the interviews revealed neutral or negative perceptions of this relationship, as well as expressions of grief following the loss of an animal and indignation toward situations of mistreatment. Taken together, the results suggest that although dogs are widely viewed in a positive light and often regarded as family members, divergent understandings of their value and roles persist, reflecting cultural tensions that influence both human and animal well-being. Full article
(This article belongs to the Section Human-Animal Interactions, Animal Behaviour and Emotion)
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21 pages, 2072 KB  
Article
Analysis of Transcriptome and Differentially Expressed Genes in Chicken Primordial Germ Cells
by Anastasiia I. Azovtseva, Anna E. Ryabova, Artem P. Dysin, Grigoriy K. Peglivanyan, Natalia R. Reinbach, Alina V. Gabova, Olga Y. Barkova, Ekaterina A. Polteva and Tatiana A. Larkina
Animals 2026, 16(3), 522; https://doi.org/10.3390/ani16030522 - 6 Feb 2026
Viewed by 1543
Abstract
Achieving successful primordial germ cell (PGC)-based genome editing requires a deep understanding of their molecular identity. For the first time, a comparative transcriptomic analysis of chicken PGCs and adult liver cells to define their specific gene expression signature was performed. PGCs were isolated [...] Read more.
Achieving successful primordial germ cell (PGC)-based genome editing requires a deep understanding of their molecular identity. For the first time, a comparative transcriptomic analysis of chicken PGCs and adult liver cells to define their specific gene expression signature was performed. PGCs were isolated from Rhode Island Red chicken embryos, cultured, and subjected to RNA sequencing alongside liver tissue. Differential expression analysis with Benjamini–Hochberg correction identified 1909 differentially expressed genes (DEGs). Functional annotation revealed that PGCs possess a unique transcriptional landscape, characterized not only by enhanced proliferation and metabolic activity but also by a profound molecular convergence with neural crest cells. This is evidenced by the upregulation of gene modules governing long-range migration, neuronal signaling, and specialized “neuro-lipid” metabolism (e.g., sphingolipid and plasmalogen pathways). Additionally, we identified unannotated transcripts linked to immune pathways and ciliary signaling. Our study expands the functional annotation of avian PGCs and reveals an unexpected evolutionary recruitment of conserved morphogenetic programs, providing a refined molecular foundation for advanced germline editing technologies. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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21 pages, 15611 KB  
Article
Seasonal Change in Glucose Metabolism and Steroidogenesis in the Ovaries of Wild Ground Squirrels (Spermophilus dauricus)
by Zhaomei Dong, Qingjing Gao, Pengyu Chen, Xi Wang, Yuning Liu, Haolin Zhang, Zhengrong Yuan, Yingying Han and Qiang Weng
Animals 2026, 16(3), 521; https://doi.org/10.3390/ani16030521 - 6 Feb 2026
Viewed by 982
Abstract
Precise regulation of ovarian glucose metabolism and steroidogenesis is pivotal for the reproductive success of seasonal breeders. This study investigated seasonal variations in ovarian glucose metabolism and its association with follicular development and steroidogenesis in wild ground squirrels (Spermophilus dauricus). Results [...] Read more.
Precise regulation of ovarian glucose metabolism and steroidogenesis is pivotal for the reproductive success of seasonal breeders. This study investigated seasonal variations in ovarian glucose metabolism and its association with follicular development and steroidogenesis in wild ground squirrels (Spermophilus dauricus). Results showed that ovarian weight and volume, and serum levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17β-estradiol, and progesterone were significantly higher in the breeding season (BS) than in the non-breeding season (NBS). Ovarian glycogen content was elevated in the BS, whereas circulating glucose levels decreased remarkably. Histologically, the BS ovaries contained follicles at all developmental stages (primary to Graafian) and corpora lutea, while corpora lutea were absent in the NBS. Integrated transcriptomic and metabolomic analyses revealed upregulated glycogen synthesis, glycolytic pathways, and altered steroidogenic metabolites in the BS. Quantitative real-time PCR (qRT-PCR) confirmed higher expression of glucose transporter 1 (GLUT1), glucose metabolism-related enzymes (G6PD, PFKFB3, PFKM), glycogen synthase (GYS), and steroidogenic factors (StAR, P450scc, P450c17, 3β-HSD, P450arom) in the BS, whereas glycogen synthase kinase-3β (Gsk-3β) was upregulated in the NBS. Immunohistochemistry demonstrated colocalization of steroidogenic and glucose-metabolizing enzymes in granulosa and theca cells, and correlation analyses confirmed positive associations between glucose metabolism, steroidogenesis, and ovarian weight. Collectively, these findings indicate that enhanced ovarian glucose metabolism in the BS is tightly coupled with elevated steroidogenesis, synergistically promoting seasonal follicular development and ovulatory competence in this species. Full article
(This article belongs to the Special Issue Advances in the Reproduction of Wild and Exotic Animals)
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19 pages, 630 KB  
Review
Nitrogen Excretion, Ammonia, and Greenhouse Gases Emission in Italian Heavy Pigs: The Role of Feed in Environmental Impact Mitigation
by Raffaella Rossi, Eleonora Buoio, Edda Mainardi and Annamaria Costa
Animals 2026, 16(3), 520; https://doi.org/10.3390/ani16030520 - 6 Feb 2026
Viewed by 1267
Abstract
This review examined the environmental impact of fattening pigs, with particular focus on the Italian heavy fattening pig, affected by dietary interventions aimed at mitigating nitrogen excretion and gases emission into the atmosphere, maintaining product quality and complying with the regulation frameworks. In [...] Read more.
This review examined the environmental impact of fattening pigs, with particular focus on the Italian heavy fattening pig, affected by dietary interventions aimed at mitigating nitrogen excretion and gases emission into the atmosphere, maintaining product quality and complying with the regulation frameworks. In the Italian heavy pig supply chain, environmental regulations are often difficult to comply with due to the constraints imposed by PDO. The pig sector is increasingly committed to developing strategies that can effectively mitigate its environmental impacts. In intensive pig farming, emissions of ammonia (NH3) and greenhouse gases (GHGs) can occur in every production phase, from animal housing to manure treatment, management, storage, and up to in-field application. All these phases present key actions to limit emissions of NH3 and GHG, according to Best Available Techniques (BAT) for housing and Best Practices (BP) for manure treatments and management. Nitrogen excretion in manure is another important aspect to consider for its negative effects when applied in Vulnerable Zones with considerable additions of animal waste and synthetic fertilisers. Nutritional strategies can contribute a priori to mitigate nitrogen excretion, NH3 emissions, and overall GHG output in heavy pigs, particularly in Italian PDO ham systems. While these interventions effectively mitigate NH3 and influence manure-related emissions, their impact on total GHG varies depending on diet composition, fibre type, additive combination, and post-excretion treatment. Full article
(This article belongs to the Section Animal Nutrition)
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18 pages, 1309 KB  
Article
Comparing Macroscopic and Quantitative Histological Methods to Determine Sexual Maturity in the Female European Plaice, Pleuronectes platessa Linnaeus, 1758
by Carine Sauger, Jérôme Quinquis, Kristell Kellner, Clothilde Berthelin, Mélanie Lepoittevin, Nicolas Elie and Laurent Dubroca
Animals 2026, 16(3), 519; https://doi.org/10.3390/ani16030519 - 6 Feb 2026
Cited by 2 | Viewed by 983
Abstract
Two methods (macroscopic and quantitative histology) to determine sexual maturity are described in this paper, with an emphasis on the methods used in stereology. Based on a point counting grid (Glagolev’s method), ovarian cellular structures were quantified throughout the whole cross section of [...] Read more.
Two methods (macroscopic and quantitative histology) to determine sexual maturity are described in this paper, with an emphasis on the methods used in stereology. Based on a point counting grid (Glagolev’s method), ovarian cellular structures were quantified throughout the whole cross section of 151 Pleuronectes platessa individuals. The novelty of this paper is checking the cellular homogeneity in the anterior, median and posterior sections of both ovaries from 15 fish, accompanied by a reading protocol to limit subjectivity during stereology readings. Once the ovarian structures were quantified for 151 individuals, a model was set up following the standardized staging grid of the International Council for the Exploration of the Sea (ICES), classifying each sampled individual into a maturity phase. These results showed that 23.4% of the time immature (A) individuals were correctly identified through the macroscopic method, but 47.9% of developing (B) individuals were classified under A, while individuals that had finished spawning were correctly identified as such 80.4% of the time. Finally, the maturity ogive and the length at which 50% of the population is sexually mature (L50) were calculated, with an L50 of 28.6 cm when using the macroscopic method and an L50 of 20.6 cm with the stereological method. Full article
(This article belongs to the Section Animal System and Management)
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14 pages, 2360 KB  
Article
Stress Responses in Dressage Horses: Insights from FEI Noseband Measurements Across National Competition Levels
by Simona Fialová, Dana Kuřitková and Eva Sobotková
Animals 2026, 16(3), 518; https://doi.org/10.3390/ani16030518 - 6 Feb 2026
Cited by 1 | Viewed by 2271
Abstract
This pilot study investigated stress-related behaviors in 238 dressage horse–rider combinations competing at national levels from Elementary (A) to Grand Prix (GP). Noseband tightness was assessed on-site using the FEI noseband measuring device (FNMD), and all but two horses complied with FEI regulations. [...] Read more.
This pilot study investigated stress-related behaviors in 238 dressage horse–rider combinations competing at national levels from Elementary (A) to Grand Prix (GP). Noseband tightness was assessed on-site using the FEI noseband measuring device (FNMD), and all but two horses complied with FEI regulations. Video-based analysis quantified conflict behaviors including mouth opening, tail swishing, and changes in head–neck position. Because noseband tightness showed minimal variation, no meaningful association with behavioral expression could be determined. In contrast, competition level had a clear influence on both the frequency and type of stress-related behaviors. Horses in lower-level classes displayed a wide range of behaviors at relatively low frequency, while horses in higher-level tests showed fewer behavior types but exhibited them more often, with mouth opening becoming the predominant indicator. Horses ridden in a double bridle generally demonstrated higher proportions of conflict behavior than those ridden in a snaffle. A strong negative correlation between conflict behavior and performance scores was observed only at the lowest levels (A, L). From Medium level upward, judging scores no longer reflected the amount of stress behavior displayed. Overall, the findings indicate that increasing test difficulty is associated with a rise in stress-related behaviors and a narrowing of behavioral expression. These results highlight the importance of considering equipment choice and competition demands when evaluating stress in dressage horses. Full article
(This article belongs to the Section Animal Ethics)
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16 pages, 4395 KB  
Article
Phylogeographic Structure Reveals Hidden Diversity Patterns in Tor tambra (Cyprinidae) Across Thai River Systems
by Vatthanachai Phanklam, Sommai Janekitkarn, Kathathep Seesan, Narongrit Muangmai, Cong Zeng, Prapansak Srisapoome, Kornsorn Srikulnath, Ling Wu, Xiao-Yong Chen, Uthairat Na-Nakorn and Ishwar S. Parhar
Animals 2026, 16(3), 517; https://doi.org/10.3390/ani16030517 - 6 Feb 2026
Cited by 1 | Viewed by 1641
Abstract
Understanding patterns of genetic diversity and population structure in freshwater fishes is crucial for evolutionary studies and conservation planning. Tor tambra is a common freshwater fish species in Thai waters, yet its genetic diversity and population connectivity remain largely unexplored. This study investigated [...] Read more.
Understanding patterns of genetic diversity and population structure in freshwater fishes is crucial for evolutionary studies and conservation planning. Tor tambra is a common freshwater fish species in Thai waters, yet its genetic diversity and population connectivity remain largely unexplored. This study investigated the phylogeographic structure and genetic diversity of Thai T. tambra through phylogenetic analyses and molecular approaches using mitochondrial DNA markers (COI and Cytb). Our analyses revealed four distinct genetic lineages (clades A-D) within Thai T. tambra. Lineage C comprised specimens from southern Thailand, Malaysia, Indonesia, and southern China, while lineages A, B, and D were distributed exclusively within Thailand. These Thai-endemic lineages exhibited pronounced geographic structuring across northern and western regions, with lineage A widespread, lineage B confined to northern areas, and lineage D restricted to western localities. The genetic differentiation and distribution patterns of these lineages suggest influence from the limited north–south river system connections and the biogeographic barrier posed by the Isthmus of Kra. Multiple lineages co-occurred at some locations, indicating a complex evolutionary history. These findings provide important implications for understanding the evolutionary processes and biogeographic factors shaping genetic diversity within T. tambra across the riverine systems of Thailand. Full article
(This article belongs to the Special Issue Phylogeography, Speciation, and Molecular Evolution in Freshwater Aquatic Fauna)
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22 pages, 2311 KB  
Article
Multi-Organ Transcriptomic Analysis of Greater Amberjack (Seriola dumerili) with Different Growth Rates
by Xiaoying Ru, Xiaojing Li, Yang Huang, Peipei Chen, Qiuxia Deng, Hang Li, Qibing Zhou, Haoyi Lin, Ruijuan Hao, Yongguan Liao, Jinhui Wu, Yanfei Zhao and Chunhua Zhu
Animals 2026, 16(3), 516; https://doi.org/10.3390/ani16030516 - 6 Feb 2026
Cited by 1 | Viewed by 1456
Abstract
In order to explore the main regulatory genes and related pathways of growth traits, transcriptome sequencing was performed on the hypothalamus, pituitary, and liver tissues of 12-month-old greater amberjack (Seriola dumerili) with different growth rates. In total, 504 (118 up- and [...] Read more.
In order to explore the main regulatory genes and related pathways of growth traits, transcriptome sequencing was performed on the hypothalamus, pituitary, and liver tissues of 12-month-old greater amberjack (Seriola dumerili) with different growth rates. In total, 504 (118 up- and 386 down-regulated), 556 (283 up- and 273 down-regulated), and 699 (224 up- and 475 down-regulated) differentially expressed genes (DEGs) were identified in the hypothalamus, pituitary, and liver tissues, respectively. GO and KEGG pathway analyses revealed significant differences in the expression of several genes involved in growth, metabolism, and immune-related pathways. The mRNA expression levels of genes related to growth (ghrh, ghra, igf1), cell proliferation (fgf19, fgfr4, mapk8b, map2k4b, and map4k3), and lipid metabolism (acsl5, dgat2, lipeb, cyp7a1, and fabp10a) were up-regulated in the fast-growing (FG) group, while the cartl and sst1.1 were down-regulated. Conversely, genes associated with glycolysis (fbp1a, pklr, pgm2), citrate cycle (aclya, idh1), and immune-related pathways (irf1b, cxcl10, tnfb, lysg, ifi44, mapk11, and mapk12b) were up-regulated in the slow-growing (SG) group. These findings indicate that the FG exhibited greater lipid metabolism capacity and cell proliferation ability, while the SG expended additional energy to cope with environmental stress, with hindered growth during immune response. This study enhances our understanding of the genetic mechanisms underlying differences in growth rates and provides essential gene resources for future growth-related molecular breeding programs in greater amberjack. Full article
(This article belongs to the Special Issue Advances in Research on Functional Genes and Economic Traits in Fish)
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12 pages, 2332 KB  
Article
Hepatic Expression of Growth Hormone Receptor (GHrec) and Insulin-like Growth Factor-I (IGF-I) Genes and Cellular Location of IGF-I mRNA in Diploid and Triploid Atlantic Salmon (Salmo salar) Undergoing Parr–Smolt Transformation
by Martina Bortoletti, Elisa Fonsatti, Lisa Maccatrozzo, Stefano Peruzzi, Malcolm Jobling, Marta Vascellari, Giuseppe Radaelli and Daniela Bertotto
Animals 2026, 16(3), 515; https://doi.org/10.3390/ani16030515 - 6 Feb 2026
Viewed by 1283
Abstract
The induction of triploidy, a strategy to mitigate unwanted pre-harvest sexual maturation and a genetic containment measure for escaped farmed Atlantic salmon (Salmo salar), may give rise to challenges because of the distinct environmental and dietary requirements of sterile triploid fish. [...] Read more.
The induction of triploidy, a strategy to mitigate unwanted pre-harvest sexual maturation and a genetic containment measure for escaped farmed Atlantic salmon (Salmo salar), may give rise to challenges because of the distinct environmental and dietary requirements of sterile triploid fish. Smoltification is a critical phase in the life cycle of Atlantic salmon, so knowledge about parr–smolt transformation in triploids is important for the salmon farming industry. This study covered an investigation of hepatic expression of growth hormone receptor (GHrec) and insulin-like growth factor-I (IGF-I) genes, both of which are intimately involved in the regulation of osmoregulation and growth. Additionally, hepatic presence and location of IGF-I mRNA were examined using RNAscope®, an advanced in situ hybridization technique. Triplicate groups of juvenile diploid and triploid salmon were reared at low temperature (10 °C) and fed either a standard diet or one enriched with hydrolyzed fish proteins from the start of feeding onwards. Liver samples were collected from three fish per tank each month from October to December (2454–3044 degree-days post-start feeding), the period encompassing smoltification, and hepatic expression of IGF-I and GHrec genes was quantified by real-time PCR. The results indicated that neither ploidy nor diet significantly influenced IGF-I or GHrec gene expression, suggesting that, under our conditions, triploidy and diet did not adversely affect this molecular pathway linked to growth and osmoregulation. IGF-I gene expression exhibited significant temporal variation, correlating with the progression of smoltification, while GHrec gene expression showed a similar, albeit non-significant, trend. Triploids exhibited IGF-I and GHrec gene expression patterns comparable to diploids, and both the temporal changes and lack of difference between triploids and diploids were mirrored in the quantification of IGF-I mRNA within the liver cells. The potential applicability to a commercial aquaculture setting requires further investigation. Full article
(This article belongs to the Special Issue Advances in Veterinary Anatomy: Connecting Basic Research with Clinical, Biotechnological, and Educational Innovations)
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10 pages, 3031 KB  
Article
Assessment of Femoral Trochlear Groove Depth in Canine Cadavers with Normal Stifle Joints Using Ultrasonography with the Joint Positioned at 45°, 90°, and Hyperextension, and Radiography in the Skyline Projection
by Amanda Junqueira, Maria Paula Luchi da Silva Mattos, Francine Hergemoller, Thayse Meyer, Caroline Bernardo Gusmão, Rafael Kretzer Carneiro and Márcio Poletto Ferreira
Animals 2026, 16(3), 514; https://doi.org/10.3390/ani16030514 - 6 Feb 2026
Viewed by 1292
Abstract
This prospective, analytical study aimed to measure and compare trochlear groove depth using skyline radiographic projection and ultrasonography at three stifle joint angles and four distinct anatomical regions, as well as to compare these findings with ex vivo measurements obtained using a caliper [...] Read more.
This prospective, analytical study aimed to measure and compare trochlear groove depth using skyline radiographic projection and ultrasonography at three stifle joint angles and four distinct anatomical regions, as well as to compare these findings with ex vivo measurements obtained using a caliper under the same conditions in canine cadavers without patellar luxation. A total of 67 limbs were evaluated. Ultrasonography demonstrated greater agreement with ex vivo measurements than radiography. Ultrasonographic assessment was considered appropriate for measuring trochlear groove depth with the stifle joint in maximal extension, with the transducer positioned distal to the patella, and at 90° of flexion, with the transducer positioned proximal to the patella. Full article
(This article belongs to the Special Issue Diagnostic Imaging for Companion Animals: Advances in Radiology and Ultrasound)
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17 pages, 2474 KB  
Article
The Period (per) Gene Plays an Important Role in Regulating Circadian Oscillation and Ovarian Development in the Ridgetail White Prawn, Exopalaemon carinicauda
by Caijuan Tian, Yunhan Feng, Jixuan Zhu, Xuanjian Chen, Wanxin Ma, Panpan Niu, Hao Zhong, Huan Gao and Yuquan Li
Animals 2026, 16(3), 513; https://doi.org/10.3390/ani16030513 - 6 Feb 2026
Viewed by 1054
Abstract
To explore the function of the period gene (Ec-per) in Exopalaemon carinicauda, we cloned the gene of 4611 bp with a 5′UTR of 201 bp, a 3′UTR of 813 bp, and an ORF of 3597 bp encoding 1198 [...] Read more.
To explore the function of the period gene (Ec-per) in Exopalaemon carinicauda, we cloned the gene of 4611 bp with a 5′UTR of 201 bp, a 3′UTR of 813 bp, and an ORF of 3597 bp encoding 1198 amino acids. The predicted protein includes two PAS and one PERIOD domain. qPCR analysis revealed that Ec-per was expressed across all tissues tested at different developmental stages and during both embryonic and larval stages. Moreover, Ec-per oscillated rhythmically under different conditions of light-to-dark (L:D) ratios, including continuous darkness (0 L:24 D), where changes in the photoperiod influenced amplitude and phase shifts. The knockdown of Ec-per mRNA significantly reduced the expression of the circadian-related genes timeless (tim) and cryptochrome 1 (cry1) (p < 0.05). This suggests that Ec-per is an endogenous clock gene that may participate in molecular feedback loops and synergistically regulate the circadian rhythms through interacting with tim and cry1. RNA interference of Ec-per also markedly downregulated ecdysone receptor mRNA (p < 0.05), suggesting a positive role in the ovarian development of E. carinicauda. In situ hybridization further demonstrated that Ec-per is involved in oocyte proliferation and the accumulation of exogenous nutrients. This study provides new insights for promoting ovarian development and artificial breeding in crustaceans through optimized light-cycle management. Full article
(This article belongs to the Section Aquatic Animals)
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15 pages, 14068 KB  
Article
Collagen-Based 3D Scaffolds from Sea Urchin Food Waste for Skeletal Muscle Tissue Engineering
by Eylem Emek Akyürek, Luca Melotti, Martina Erba, Anna Carolo, Giordana Martinelli, Margherita Roncoroni, Stefania Marzorati, Marco Patruno, Michela Sugni and Roberta Sacchetto
Animals 2026, 16(3), 512; https://doi.org/10.3390/ani16030512 - 5 Feb 2026
Viewed by 994
Abstract
Preclinical investigations involving in vivo animal studies, are considered a crucial and mandatory step in pharmacological industry regulations. In recent years, as the ethical concerns associated with animal experimentation are becoming more relevant, substantial research efforts have been directed towards the development of [...] Read more.
Preclinical investigations involving in vivo animal studies, are considered a crucial and mandatory step in pharmacological industry regulations. In recent years, as the ethical concerns associated with animal experimentation are becoming more relevant, substantial research efforts have been directed towards the development of novel in vitro methodological approaches aimed at minimizing the utilization of animals. Among those, three-dimensional (3D) cell culture systems have become one of the most promising substitutes for animal models. Skeletal muscle can be affected by genetic disorders and injuries. In this study, two types of 3D scaffolds (Coll and CollMA), prepared starting from a fibrillar collagen suspension extracted from sea urchin food waste, were used as platforms for the generation of 3D skeletal muscle tissue models. The scaffolds were cellularized with C2C12 myoblasts. As the culture period progressed, C2C12 cells gradually infiltrated and were viable throughout both scaffold types, establishing a multilayered cellular population. In the CollMA scaffold, the reciprocal Pax7/MyoD expression pattern suggests a progression of myoblasts towards myogenic commitment. Taken together, our results, although preliminary, suggest that sea urchin–derived collagen matrix represents a promising scaffold for skeletal muscle tissue engineering. Full article
(This article belongs to the Special Issue Feature Papers in Veterinary Clinical Studies: From Current to Emerging Technologies and Innovative Approaches for the Future)
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15 pages, 6502 KB  
Article
Molecular Cloning and Expression Responses to Streptococcus agalactiae and Aeromonas veronii of TLR19, TLR20, and TLR21 in Schizothorax prenanti
by Qiyu Luo, Jie Zhang, Yao Shi, Yanjing Zhao, Yuanchao Zou and Xianghui Kong
Animals 2026, 16(3), 511; https://doi.org/10.3390/ani16030511 - 5 Feb 2026
Viewed by 1065
Abstract
Toll-like receptors (TLRs) are essential pattern recognition receptors of the innate immune system and play critical roles in pathogen invasion in teleosts. In this study, we identified and characterized full-length open reading frames of three TLRs belonging to the TLR11 subfamily from Schizothorax [...] Read more.
Toll-like receptors (TLRs) are essential pattern recognition receptors of the innate immune system and play critical roles in pathogen invasion in teleosts. In this study, we identified and characterized full-length open reading frames of three TLRs belonging to the TLR11 subfamily from Schizothorax prenanti, termed spTLR19 (2868 bp), spTLR20 (2835 bp), and spTLR21 (2946 bp), encoding 955, 944, and 981 amino acids, respectively. All three proteins exhibited the conserved domain architecture typical of TLRs, comprising a leucine-rich repeat (LRR) domain, a transmembrane region, and a Toll/IL-1 receptor (TIR) domain. Phylogenetic and homology analyses revealed that spTLR19 and spTLR20 clustered most closely with their homologues from Cyprinus carpio, while spTLR21 showed the highest similarity to Onychostoma macrolepis TLR21. Expression profiling showed that these TLRs were ubiquitously expressed across examined tissues, with relatively higher expression in immune-related tissues such as spleen and gills. Furthermore, challenge with Streptococcus agalactiae and Aeromonas veronii significantly up-regulated the expression of spTLR19, spTLR20, and spTLR21 in spleen, liver, and gills, suggesting their involvement in antibacterial immune responses. These findings enhance the functional understanding of the teleost TLR11 subfamily and provide a foundation for elucidating disease resistance and immune regulation in S. prenanti. Full article
(This article belongs to the Section Aquatic Animals)
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17 pages, 2441 KB  
Article
Rumen Microbial Composition and Fermentation Variables Associated with Methane Production in Italian Simmental Dairy Cows
by Cristina Pavanello, Marcello Franchini, Alberto Romanzin, Lara Tat, Stefano Bovolenta and Mirco Corazzin
Animals 2026, 16(3), 510; https://doi.org/10.3390/ani16030510 - 5 Feb 2026
Cited by 1 | Viewed by 1168
Abstract
The study investigated differences in ruminal and fecal microbiota composition, fermentation traits, and volatile organic compounds (VOC) in Simmental dairy cows classified as high (HME) or low (LME) methane emitters. Methane emissions from 48 cows were quantified using the Laser Methane Smart portable [...] Read more.
The study investigated differences in ruminal and fecal microbiota composition, fermentation traits, and volatile organic compounds (VOC) in Simmental dairy cows classified as high (HME) or low (LME) methane emitters. Methane emissions from 48 cows were quantified using the Laser Methane Smart portable gas detector. The 12 animals with the highest and lowest emissions were selected and assigned to the HME and LME groups, respectively, balanced for body weight, days in milk, and body condition score. Rumen fluid and fecal samples were analyzed for pH, ammonia, volatile fatty acids (VFA), VOC, and microbiota composition. As expected, CH4 emissions were significantly higher in HME than in LME cows (22.5 vs. 13.2 g/kg DMI; 16.9 vs. 8.4 g/kg FCM). The neutral detergent fiber digestibility was higher in HME cows (51.4% vs. 47.9%). The valeric acid concentration and the acetate-to-propionate ratio were significantly higher in HME cows (3.53 vs. 3.31). The VOC profiles significantly differed between groups in both feces and rumen fluid. The microbiota analysis revealed a significant difference between groups at the order and genus levels (Bray–Curtis dissimilarity). The Shannon index was higher in LME cows (2.08 vs. 1.95). HME cows exhibited a higher abundance of Methanosphaera and Methanobacteriales. Overall, the results indicate that re-shaping the rumen microbial community can play a key role in reducing methane emissions, strengthening the case for microbiome-driven approaches and offering insights that can support mitigation strategies across dairy production systems. Full article
(This article belongs to the Section Cattle)
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19 pages, 2740 KB  
Article
Genomic Signatures Underlying Environmental Adaptation and Reproductive Traits in the Tibetan Pig
by Mengqi Duan, Songyuan Zhang, Hang Jiao, Peng Shang, Chunli Li and Kejun Wang
Animals 2026, 16(3), 509; https://doi.org/10.3390/ani16030509 - 5 Feb 2026
Viewed by 1273
Abstract
Background: The Tibetan pig, a highland breed with exceptional adaptability to harsh environments (cold, hypoxia, coarse feed) but poor growth/reproductive traits, was studied to uncover genetic mechanisms and support breeding improvements. Methods: We conducted de novo genome assembly of a male Tibetan pig [...] Read more.
Background: The Tibetan pig, a highland breed with exceptional adaptability to harsh environments (cold, hypoxia, coarse feed) but poor growth/reproductive traits, was studied to uncover genetic mechanisms and support breeding improvements. Methods: We conducted de novo genome assembly of a male Tibetan pig using stLFR sequencing, supplemented with ONT data, and compared the assembly to the Duroc pig genome (v11.1). Results: The assembled genome (2.25 Gb, contig N50 = 136.5 Mb, GC content = 41.74%, 94.16× coverage) showed 96.9% BUSCO completeness. Structural variant (SV) analysis identified 22,008 insertions and 27,639 deletions, with an SV genotyping accuracy of 0.9735. Selective sweep analysis highlighted adaptive genes: XIRP2 (cardiac function), KSR2/CACNA1A (fat metabolism), COL11A1 (cartilage), and ADORA2A (vascular regulation). Tibetan pigs exhibited the fewest and shortest runs of homozygosity (ROHs) among four breeds, with ROH-linked SNPs implicating lipid catabolism genes (LIPE, PNPLA2, MGLL, DGAT1). An SNP-based GWAS revealed reproductive trait associations: immune gene IL2RB, energy metabolism genes PRKAG2, ADGRA1, and PTPRN2, and growth genes SLIT2 and BMP6. SV analysis identified additional candidates: energy metabolism genes HAO2 and NRG4, growth genes MTUS2 and FGF12, and immune genes SCGB1A1 and C8A. Conclusions: This study provides a chromosome-level genome assembly of a male Tibetan pig (generated from stLFR and ONT data), and, through whole-genome resequencing of 124 Tibetan sows, identifies key genetic factors underlying Tibetan pigs’ environmental adaptability and reproductive limitations, enabling genomic strategies to enhance breeding efficiency while preserving adaptive traits. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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18 pages, 3964 KB  
Article
Dynamic Metabolic Changes During Postmortem Aging of Yili Horsemeat Revealed by Untargeted Metabolomics
by Xixi Yang, Tongliang Wang, Jingtao Gan, Chen Meng, Jingxuan Shen, Zexu Li, Xueyan Li, Yaqi Zeng, Wanlu Ren, Xinkui Yao and Jun Meng
Animals 2026, 16(3), 508; https://doi.org/10.3390/ani16030508 - 5 Feb 2026
Viewed by 1140
Abstract
Postmortem aging is a crucial process for improving meat quality; however, the metabolic mechanisms underlying the postmortem maturation of Yili horsemeat remain unclear. In this study, untargeted metabolomics was employed to investigate the dynamic changes in metabolites during postmortem aging of Yili horse [...] Read more.
Postmortem aging is a crucial process for improving meat quality; however, the metabolic mechanisms underlying the postmortem maturation of Yili horsemeat remain unclear. In this study, untargeted metabolomics was employed to investigate the dynamic changes in metabolites during postmortem aging of Yili horse longissimus dorsi muscle. Samples from six Yili horses were subjected to aging at 4 °C for 0, 14, and 28 days. Intramuscular Environment and metabolic profiles were analyzed using UHPLC-QTOF-MS. Results showed that meat pH initially decreased and then increased over the aging period (p < 0.05), whereas muscle fiber diameter and cross-sectional area gradually declined (p < 0.05). Metabolomic analysis revealed significant differences in metabolic profiles among different aging stages. KEGG pathway enrichment analysis indicated that the differentially expressed metabolites (DEMs) were predominantly involved in purine metabolism, nucleotide metabolism, and amino acid biosynthesis pathways. Notably, the degradation of AMP and IMP into inosine and hypoxanthine and the accumulation of free amino acids such as tryptophan and phenylalanine were closely associated with the development of umami taste and overall flavor of horsemeat. These findings elucidate the key metabolic pathways regulating postmortem meat quality in Yili horses and provide a theoretical basis for its production and application. Full article
(This article belongs to the Section Equids)
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21 pages, 4085 KB  
Article
Molecular Prevalence of Hemotropic Mycoplasmosis and Associated Risk Factors for Co-Infection with Gastrointestinal Nematode in Anemic Meat Goats of Northeast Thailand
by Sarinya Rerkyusuke, Chariya Promphak, Pattiya Wongpattaraworakul, Pimchanok Taikitsayakun, Warisa Chuduang, Thakorn Thanaakkarasophon, Worakamol Chonsirikraisri, Julamanee Suriyapoom, Suthida Chanlun, Prapan Kaenjampa, Sawarin Lerk-u-suke, Peerapol Sukon and Patchara Phuektes
Animals 2026, 16(3), 507; https://doi.org/10.3390/ani16030507 - 5 Feb 2026
Viewed by 1125
Abstract
Hemotropic mycoplasmosis is an emerging vector-borne infection in goats caused by hemotropic mycoplasmas, which leads to anemia, poor growth, abortion, and reduced productivity. In Northeastern Thailand, smallholder meat goat systems expose animals to blood-feeding vectors and gastrointestinal parasites; however, epidemiological data on these [...] Read more.
Hemotropic mycoplasmosis is an emerging vector-borne infection in goats caused by hemotropic mycoplasmas, which leads to anemia, poor growth, abortion, and reduced productivity. In Northeastern Thailand, smallholder meat goat systems expose animals to blood-feeding vectors and gastrointestinal parasites; however, epidemiological data on these issues are limited. This study examined the prevalence of hemotropic mycoplasmosis, its co-infection with gastrointestinal parasites, and the associated risk factors in anemic goats. PCR and sequence analyses confirmed the presence of Mycoplasma ovis, Candidatus Mycoplasma haematobovis and Candidatus Mycoplasma haematovis, with herd-level and individual-level prevalences of all hemotropic mycoplasmas (HMs) at 93.3% (14/15; 95% CI: 68.1–99.8%) and 59.8% (52/87; 95% CI: 48.7–70.1%), respectively. Ca. M. haematobovis was identified as the predominant species. Multivariable analysis indicated that age ≥ 1 year is a significant risk factor for HM infection (adjusted OR: 9.88; 95% CI: 1.73–56.48; p = 0.01). Co-infection between HM and gastrointestinal nematodes was found to be associated with age (p < 0.05). Effective management requires targeted selective treatment, post-treatment monitoring of infection status, vector control, and farmer education on zoonotic risks. These findings provide critical insights for designing evidence-based surveillance, control, and prevention strategies to improve goat health and productivity in smallholder tropical systems. Full article
(This article belongs to the Section Small Ruminants)
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15 pages, 1336 KB  
Article
Dietary Cerium (Ammonium Ceric Nitrate) Promoted the Growth, Intestinal Digestive Enzyme Activity, and Positive Modulation of Intestinal Microbiota of Largemouth Bass (Micropterus salmoides)
by Yugui Zhang, Yunfeng Chen, Kaihui Xu, Xiaoqin Li and Xiangjun Leng
Animals 2026, 16(3), 506; https://doi.org/10.3390/ani16030506 - 5 Feb 2026
Viewed by 906
Abstract
This study investigated the dietary effects of cerium (ammonium ceric nitrate, Ce (NH4)2(NO3)6) on the growth, serum antioxidant, intestinal digestive enzyme activity, tissue morphology and microbiota of Micropterus salmoides. Seven diets were designed with [...] Read more.
This study investigated the dietary effects of cerium (ammonium ceric nitrate, Ce (NH4)2(NO3)6) on the growth, serum antioxidant, intestinal digestive enzyme activity, tissue morphology and microbiota of Micropterus salmoides. Seven diets were designed with cerium supplementation of 0 (CON), 10 (Ce10), 20 (Ce20), 40 (Ce40), 60 (Ce60), 80 (Ce80) and 120 mg/kg (Ce120), respectively. Largemouth bass juveniles (initial weight of 16.89 ± 0.04 g) were fed with the above diets for 56 days. Compared with the control group, the weight gain of the Ce40 group increased by 14.4% and the feed conversion ratio decreased by 0.13 (p < 0.05). The Ce60, Ce80 and Ce120 groups showed significantly higher superoxide dismutase activity and lower malondialdehyde concentration compared with the control group (p < 0.05). Protease activity in the Ce20 and Ce40 groups and amylase activity in the Ce40 group were markedly elevated relative to the control group (p < 0.05). The proportion of Firmicutes was increased and the proportion of Proteobacteria was decreased by the addition of 10 mg/kg and 40 mg/kg Ce (Ce10 and Ce40 groups). In summary, dietary cerium supplementation can promote the growth, intestinal digestive enzyme activity, and positive modulation of the intestinal microbial flora of juvenile Micropterus salmoides. Based on the second-order polynomial regression analysis of WG or the FCR, the appropriate inclusion level of dietary cerium for juvenile largemouth bass was estimated to be 57.9 and 60.0 mg/kg, respectively. Full article
(This article belongs to the Section Animal Nutrition)
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7 pages, 276 KB  
Opinion
When Pain Outlasts Our Drugs: The Case for Sustained-Release Analgesia in Sheep
by Mahla Nateghi Baygi, Lee Narelle Metcalf, Benjamin Kimble and Sabrina Lomax
Animals 2026, 16(3), 505; https://doi.org/10.3390/ani16030505 - 5 Feb 2026
Viewed by 1402
Abstract
Pain associated with routine husbandry procedures in sheep can persist for several days or even weeks, yet current analgesic options, such as the non-steroidal anti-inflammatory drug (NSAID) meloxicam, typically provide only 24–36 h of analgesia. This mismatch between pain duration and analgesic coverage [...] Read more.
Pain associated with routine husbandry procedures in sheep can persist for several days or even weeks, yet current analgesic options, such as the non-steroidal anti-inflammatory drug (NSAID) meloxicam, typically provide only 24–36 h of analgesia. This mismatch between pain duration and analgesic coverage represents a fundamental limitation of current pain-management strategies in sheep. Sustained-release (SR) formulations are emerging as a promising approach to deliver longer-lasting pain relief from a single dose, reducing the need for repeated handling, and improving both animal welfare and farm efficiency. Emerging evidence highlights both the promise and limitations of different strategies to extend therapeutic coverage beyond 72 h. While preliminary results are encouraging, challenges remain in achieving consistent pharmacokinetic profiles and optimal peak concentrations. Advancing SR meloxicam formulations could support widespread uptake of welfare-focused innovations in the sheep industry. Although not yet widely available, sustained-release meloxicam represent a promising step towards making routine husbandry procedures more humane and efficient. Full article
(This article belongs to the Section Veterinary Clinical Studies)
20 pages, 2393 KB  
Article
Anti-Inflammatory Effects of Lipoxin A4 in Salmonella Typhimurium-Induced Enteritis in Wenchang Chickens
by Xiaoxiao Li, Hesi Ma, Jiankun Huang, Xuhua Ran and Xiaobo Wen
Animals 2026, 16(3), 504; https://doi.org/10.3390/ani16030504 - 5 Feb 2026
Viewed by 1065
Abstract
S. Typhimurium infection has the capacity to elicit enteric inflammation and metabolic dysfunction among poultry. Prior research conducted by our laboratory observed an increase in LXA4 titers within the gut of Wenchang chickens following infection with S. Typhimurium. Based on this observation, [...] Read more.
S. Typhimurium infection has the capacity to elicit enteric inflammation and metabolic dysfunction among poultry. Prior research conducted by our laboratory observed an increase in LXA4 titers within the gut of Wenchang chickens following infection with S. Typhimurium. Based on this observation, the present study analyzed the changes in body weight, immune organ indices, the levels of intestinal inflammatory cytokines, as well as cyclooxygenase-2 (COX-2) expression in Wenchang chickens before and after infection. The findings indicated that S. Typhimurium infection led to reduced body weight and significantly decreased thymus and bursa indices. Furthermore, a significant elevation was observed in the transcript levels of pro-inflammatory mediators, including IL-1β, along with IL-6, and TNF-α, concurrently with an increase in the mRNA transcript levels of the enzyme COX-2. Treatment with LXA4 attenuated these alterations and effectively alleviated the inflammatory response. Additionally, an in vitro system was employed to validate the anti-inflammatory properties of LXA4 against S. Typhimurium-induced inflammation in chicken HD11 macrophages. The results demonstrated that LXA4 attenuated the transcript levels of IL-1β, as well as IL-6, TNF-α, and COX-2, at various intervals (2, 12, and 24 h), thereby alleviating inflammation elicited by S. Typhimurium challenge. We employed the LXA4 receptor antagonist Boc-2 to explore the ALX/FPR2 signaling axis and noted the successful neutralization of LXA4-mediated anti-inflammatory properties by this antagonist in S. Typhimurium–challenged HD11 macrophages. Collectively, these findings indicate that S. Typhimurium triggers pro-inflammatory reactions across both in vivo chicken models and in vitro HD11 macrophage systems, whereas LXA4 effectively mitigates this inflammatory process. This research establishes the conceptual underpinnings necessary to advance the design of therapeutic modalities aimed at counteracting S. Typhimurium challenges within poultry populations. Full article
(This article belongs to the Section Animal Physiology)
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16 pages, 264 KB  
Article
Full Replacement of Soybean Meal with Soybean Press Cake in Diets of Pigs Intended for Long-Cured Dry Ham Production
by Luca Sardi, Simona Belperio, Giovanna Martelli and Eleonora Nannoni
Animals 2026, 16(3), 503; https://doi.org/10.3390/ani16030503 - 5 Feb 2026
Viewed by 1036
Abstract
The study investigated whether the full replacement of chemically solvent-extracted soybean meal (SM) with mechanically extracted soybean press cake (SC) could be carried out in pigs intended for dry-cured ham production. Eighty Italian heavy pigs were fattened and finished up to a body [...] Read more.
The study investigated whether the full replacement of chemically solvent-extracted soybean meal (SM) with mechanically extracted soybean press cake (SC) could be carried out in pigs intended for dry-cured ham production. Eighty Italian heavy pigs were fattened and finished up to a body weight of 165 kg, with half being fed a commercial diet containing SM and the other half fed an isoenergetic and isonitrogenous diet in which SC fully replaced SM. Growth parameters, carcass traits, fresh meat and cured ham quality, and fatty acid composition were analyzed. Thighs were processed according to the Parma ham production rules, which require a long curing period. The only noteworthy differences observed concerned an increase in polyunsaturated fatty acid levels in the subcutaneous fat of the SC group compared with the SM group (17.26 vs. 14.58%, p < 0.05, in fresh thighs and 15.83 vs. 12.14%, p < 0.01, in cured hams), and particularly in linoleic acid (14.85 vs. 13.01%, p < 0.01 in fresh thighs and 13.72 vs. 10.64%, p < 0.01 in cured hams), which is consistent with general nutritional recommendations favoring unsaturated over saturated fatty acids. These modifications did not affect the final quality, oxidative stability, or visual appearance of the long-cured hams. In conclusion, soybean cake–based diets can be applied throughout the production cycle of heavy pigs; however, higher inclusion levels should be carefully managed, as they may affect parameters related to suitability for dry curing. Full article
(This article belongs to the Special Issue Featured Papers in the 'Animal Products' Section)
21 pages, 840 KB  
Article
Use of Animal-Derived Products for Medicinal and Belief-Based Purposes in Urban Cities of Southwestern Nigeria: A One Health Perspective
by Samuel N. Akpan, Ralph Buij, Frank van Langevelde, Lian F. Thomas, Ayotunde E. Sijuwola, Olusola A. Ogunsanya, Pim van Hooft, Oluwatobi A. Adedokun, Abraham A. Adeyemo, Akeemat O. Ayinla, Dawn M. Zimmerman, Elizabeth A. J. Cook, Sherril P. Masudi, James M. Hassell, Christian T. Happi and Anise N. Happi
Animals 2026, 16(3), 502; https://doi.org/10.3390/ani16030502 - 5 Feb 2026
Viewed by 1680
Abstract
Zootherapy is a significant component of traditional medicine in many parts of the world. This study investigated the therapeutic and belief-based use of wildlife in urban settings in Southwest Nigeria, assessing its potential implications for One Health. We used semi-structured questionnaires, focus group [...] Read more.
Zootherapy is a significant component of traditional medicine in many parts of the world. This study investigated the therapeutic and belief-based use of wildlife in urban settings in Southwest Nigeria, assessing its potential implications for One Health. We used semi-structured questionnaires, focus group discussions, and participant observations to gain insights into the practices, practitioners’ knowledge, motivations, and animal species used. Also, we searched the published literature and the International Union for Conservation of Nature Red List of Threatened Species for zoonotic pathogens and the conservation statuses of the reported animal species. Descriptive statistics and thematic analysis were used to analyze the data. Results revealed 53 practices involving 41 species, with occasional importation of animal parts to meet demand. Practitioners’ limited awareness of zoonotic risks, coupled with income generation and deeply rooted traditional beliefs, sustained these practices, further fueled by the dynamics of urban wild meat trade. The majority (95%) of species reported were reservoirs of zoonotic pathogens, with 44% in the vulnerable, near-threatened, endangered, or critically endangered class. The use of animal-derived products for medicinal and spiritual purposes may constitute a plausible pathway for potential zoonotic spillovers, transboundary animal diseases (TADs), decline in wildlife resources, and ecological imbalance. We recommend intensified monitoring, risk communication, awareness programs, and the adoption of sustainable alternatives to mitigate the negative impacts of these practices in Nigeria and beyond. Full article
(This article belongs to the Section Animal Products)
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20 pages, 1310 KB  
Article
Antimicrobial Resistant Isolates in Surgical and Bite Wounds in Dogs and Cats: A 12-Year Retrospective Analysis
by Davide Danieli, Michela Amadori, Sara Crimi, Federica Pregnolato, Chiara Caruso, Graziana Gambino, Giovanni Re and Cristina Vercelli
Animals 2026, 16(3), 501; https://doi.org/10.3390/ani16030501 - 5 Feb 2026
Viewed by 1166
Abstract
Antimicrobial resistance (AMR) is an increasingly concern in both human and veterinary medicine, complicating the management of common clinical infections. Surgical site infections (SSIs) and bite wounds in pets are especially challenging due to their polymicrobial nature and multidrug-resistant strains. Laboratory records from [...] Read more.
Antimicrobial resistance (AMR) is an increasingly concern in both human and veterinary medicine, complicating the management of common clinical infections. Surgical site infections (SSIs) and bite wounds in pets are especially challenging due to their polymicrobial nature and multidrug-resistant strains. Laboratory records from a referral veterinary hospital in northwestern Italy were retrospectively analyzed. Bacterial cultures and antimicrobial susceptibility testing results from canine and feline surgical and bite wounds collected between 2013 and 2024 were reviewed. Data were analyzed descriptively to identify bacterial distribution and resistance trends. A total of 35 isolates (23 dogs; 12 cats) were obtained from 26 surgical and 9 bite wounds. In dogs, Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, and Staphylococcus pseudintermedius were most frequently identified. In cats, Enterobacter cloacae, Escherichia coli, and Pasteurella multocida predominated. High resistance rates were recorded for amoxicillin–clavulanate, cephalexin, enrofloxacin, and marbofloxacin. Methicillin resistance emerged in most Staphylococcus aureus and several S. pseudintermedius isolates, while subsets of E. coli and K. pneumoniae were ESBL. Resistance to carbapenems remained low. Although samples were limited, these findings underscore the ongoing impact of AMR in surgical and bite wounds and highlight the importance of implementing rigorous antimicrobial stewardship practices in veterinary medicine. Full article
(This article belongs to the Special Issue Antimicrobial Resistance and Innovative Strategies in Veterinary Medicine)
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24 pages, 394 KB  
Article
Milk Yield and Quality, Metabolic Profile and Oxidative Status in Lactating Goats, as Affected by Silage Based on Agro-Industrial By-Products
by Angela Gabriella D’Alessandro, Abdelfattah Z. M. Salem and Giovanni Martemucci
Animals 2026, 16(3), 500; https://doi.org/10.3390/ani16030500 - 5 Feb 2026
Cited by 2 | Viewed by 1249
Abstract
Large amounts of agro-industrial residues—such as grape pomace, olive mill wastewater, wheat straw, and cheese whey—pose disposal challenges and generate substantial environmental and economic burdens. Incorporating these by-products into ruminant diets may reduce feeding costs while supporting sustainability within a circular bioeconomy. Two [...] Read more.
Large amounts of agro-industrial residues—such as grape pomace, olive mill wastewater, wheat straw, and cheese whey—pose disposal challenges and generate substantial environmental and economic burdens. Incorporating these by-products into ruminant diets may reduce feeding costs while supporting sustainability within a circular bioeconomy. Two experiments were conducted to evaluate: (i) the characteristics of a mixed raw agro-industrial by-product silage (BPS) in cylindrical bale silos (50 kg) and (ii) its effects as dietary supplementation on metabolic profile, oxidative status, milk yield, and milk quality in lactating goats. The BPS was formulated from raw wheat straw, grape pomace, olive mill wastewater, and cheese whey and tested at a 30% inclusion level (dry-matter basis). The combined by-products produced in 50 kg cylindrical bale silos were positively characterized and exhibited a nutritionally suitable silage with relevant antioxidant potential. Dietary BPS improved oxidative status, as evidenced by increased TAS and vitamin E and reduced ROMs, and positively influenced lipid and immunological blood markers of goats. Milk yield was higher in goats receiving the BPS. Moreover, milk quality was improved through a more favorable fatty acid profile, higher vitamin E content, reduced cholesterol levels, and enhanced oxidative stability, as evidenced by lower TBARS values. Overall, BPS supplementation enhanced animal health indicators and milk nutritional properties, demonstrating that this silage can contribute to more sustainable dairy goat production while offering potential benefits for human nutrition. Full article
(This article belongs to the Section Small Ruminants)
21 pages, 3177 KB  
Article
Effects of Different Sources and Dietary Inclusion Levels of Astaxanthin on Growth Performance, Skin Pigmentation, and Physiological Parameters of Red Sea Bream (Pagrus major) Juveniles
by Arkadios Dimitroglou, Stephanie Carvajal Acevedo, Konstantina Evangelia Gleni, Athanasios Samaras, Dimitrios Barkas, Leonidas Papaharisis and Michael Pavlidis
Animals 2026, 16(3), 499; https://doi.org/10.3390/ani16030499 - 5 Feb 2026
Viewed by 1166
Abstract
Red seabream (Pagrus major) reared under intensive rearing conditions faces hypermelanosis and dyspigmentation, resulting in darker and less pink/red skin color, losing the natural appearance of the species. This has a great negative impact on consumers’ acceptance and market appeal. Two [...] Read more.
Red seabream (Pagrus major) reared under intensive rearing conditions faces hypermelanosis and dyspigmentation, resulting in darker and less pink/red skin color, losing the natural appearance of the species. This has a great negative impact on consumers’ acceptance and market appeal. Two experiments were performed to test the effect of the source, level, and feeding duration of astaxanthin supplementation in the feed on red seabream skin color reversing the depigmentation problem. Firstly, three different sources of astaxanthin (artificially synthesized, algal-extracted, and yeast-extracted astaxanthin) at the same inclusion level (100 mg kg−1 feed) were tested. Then, the best performing source was tested using five inclusion levels (0, 20, 60, 80, and 100 mg kg−1) in the feed for 60 days. At the same time, growth performance, blood biochemical parameters, antioxidant capacity and cortisol levels were also assessed. The results showed that red seabream appearance can be significantly improved with the inclusion of astaxanthin from the algae Haematococcus pluvialis in the feed even at lower levels compared to the artificially synthesized astaxanthin for a 60-day period. Fish growth performance and blood parameters were not affected by any of the experimental treatments. Full article
(This article belongs to the Section Aquatic Animals)
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9 pages, 372 KB  
Communication
Why Camera-Based and Scale-Based Measurements Differ: A Physiological Model of Diurnal Weight Variation in Finishing Pigs
by Kikuhito Kawasue, Khin Dagon Win and Tadaaki Tokunaga
Animals 2026, 16(3), 498; https://doi.org/10.3390/ani16030498 - 5 Feb 2026
Viewed by 550
Abstract
Live weight is widely used as a reference indicator for growth performance and for evaluating the accuracy of weight measurement technologies in pig production. However, live weight is not a fixed physiological quantity, and finishing pigs naturally experience substantial short-term mass fluctuations due [...] Read more.
Live weight is widely used as a reference indicator for growth performance and for evaluating the accuracy of weight measurement technologies in pig production. However, live weight is not a fixed physiological quantity, and finishing pigs naturally experience substantial short-term mass fluctuations due to normal behaviors such as drinking, feeding, urination, and defecation. In this study, we integrated published physiological and behavioral parameters into a stochastic simulation model to quantify within-day live-weight dynamics in finishing pigs weighing approximately 100 kg. The simulation was conducted with 1-min temporal resolution over a 24-h period. The model demonstrated that short-term weight fluctuations of approximately ±3–5 kg can occur within a single day, even when measurement error is minimal. Across 1000 simulated pigs, the mean daily fluctuation range was 4.2 kg, confirming that kilogram-scale variation is physiologically expected under normal conditions. These results provide a plausible physiological basis for understanding the frequently reported discrepancies between camera-based weight estimates and instantaneous floor-scale measurements. Camera systems primarily reflect body mass derived from external morphology, whereas floor scales measure instantaneous total mass that includes transient contributions from gastrointestinal contents, ingested water, and retained waste. Consequently, direct comparisons based on instantaneous scale readings can be misleading when used as ground truth. Our findings indicate that commonly cited accuracy claims of ±2–3 kg for camera weighing systems should be interpreted with caution, as normal physiological weight variation often exceeds this range. Recognizing live weight as a dynamic physiological variable is essential for developing biologically meaningful evaluation frameworks and for the appropriate interpretation and comparison of weight measurement technologies in precision livestock farming. Full article
(This article belongs to the Section Pigs)
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18 pages, 9366 KB  
Article
Gastric and Small-Intestinal Morphological Remodeling After Intragastric Apelin-13 Administration in Unweaned Rats
by Sylwia Szymańczyk, Cezary Osiak-Wicha, Katarzyna Kras, Małgorzata Kapica, Iwona Puzio, Hanna Antushevich, Atsukazu Kuwahara, Ikuo Kato, Iwona Łuszczewska-Sierakowska and Marcin B. Arciszewski
Animals 2026, 16(3), 497; https://doi.org/10.3390/ani16030497 - 5 Feb 2026
Viewed by 623
Abstract
Apelin is a postnatal peptide implicated in gastrointestinal maturation, yet its combined effects on mucosa, enteric plexuses, and gut-derived appetite signals are not well defined. We investigated the impact of chronic intragastric apelin-13 on the stomach and small intestine of unweaned rats. Twelve [...] Read more.
Apelin is a postnatal peptide implicated in gastrointestinal maturation, yet its combined effects on mucosa, enteric plexuses, and gut-derived appetite signals are not well defined. We investigated the impact of chronic intragastric apelin-13 on the stomach and small intestine of unweaned rats. Twelve Wistar pups of both sexes received apelin-13 (100 nmol/kg body weight, twice daily) or saline from postnatal day 10 for 14 days. After euthanasia, gastric and small-intestinal samples were processed for histomorphometry, neurofilament immunohistochemistry of myenteric and submucosal plexuses, and quantitative staining for ghrelin and leptin. Apelin-13 increased gastric mucosal thickness and pit and gland height, enlarged zymogen cells, and reduced muscularis propria thickness, while leaving submucosa and parietal cell area unchanged. In the small intestine, apelin produced a clear proximal-distal gradient, with enhanced villus-mucosa indices proximally and reduced indices in mid-to-distal jejunum, alongside broader crypt remodeling. Enterocyte and goblet cell dimensions changed in parallel with these regional shifts. Myenteric and submucosal ganglia were also remodeled in a segment-dependent manner. Ghrelin immunoreactivity increased in most regions, whereas leptin showed opposite proximal and distal responses. Overall, early-life luminal apelin-13 reshapes gastric and intestinal architecture and local hormone expression. Full article
(This article belongs to the Special Issue Advances in Veterinary Anatomy: Connecting Basic Research with Clinical, Biotechnological, and Educational Innovations)
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11 pages, 242 KB  
Article
In-Depth Analysis of the Prognostic Factors Associated with Short-Term Outcome in Equine Colic Patients: Multicentric Retrospective Study
by Irene Nocera, Dania Cingottini, Chiara Di Franco, Giulia Sala, Francesca Bindi, Alessandro Spadari, Riccardo Rinnovati, Valentina Vitale, Eduard Jose-Cunilleras and Micaela Sgorbini
Animals 2026, 16(3), 496; https://doi.org/10.3390/ani16030496 - 5 Feb 2026
Viewed by 1571
Abstract
Several studies investigated risk and prognostic parameters for horses with colic; however, the consensus is still debated. The present work aimed to investigate colic outcomes and to identify risk factors in horses referred for colic. In this multicenter retrospective study, 236 clinical records [...] Read more.
Several studies investigated risk and prognostic parameters for horses with colic; however, the consensus is still debated. The present work aimed to investigate colic outcomes and to identify risk factors in horses referred for colic. In this multicenter retrospective study, 236 clinical records of equids referred for colic at three different equine centers were reviewed. The following data were collected: history, signalment, physical examination at the time of admission, hematological and biochemical analysis, diagnosis, SIRS status and 0–6 point-scale SIRS score, colic type, treatment attempted, and outcome. Descriptive statistics were performed, and distribution of continuous variables was reported as median and percentile. A multivariable logistic regression model was applied to assess parameters associated with colic outcomes in horses (p < 0.05). A total of 138/236 horses were included in the study. The univariate analysis identified as potentially associated with the outcome: sex (p = 0.046), colic type (p < 0.001), treatment type (p < 0.001), SIRS score (p = 0.049), age (p-value = 0.057), heart rate (p = 0.013), and respiratory rate (p = 0.017). The logistic regression model indicated that colic type (p < 0.001) and age (p = 0.004) were significantly associated with a negative outcome. Equine colic risks are multifactorial; prognosis declines with age and strangulating obstructive non-strangulating colic. Poor outcomes link to cardiovascular signs like elevated heart rate, SIRS status and score, and blood lactate. Heterogeneity from diverse sites limits generalizability, but standardized protocols, binarized data, and a multicenter approach enhance robustness and representativeness while reducing local biases. Full article
(This article belongs to the Special Issue Advances in Diagnostic and Treatment Methods for Equine Gastrointestinal Diseases)
28 pages, 2298 KB  
Review
Effects of Thermal Stress on Growth and Reproduction of Procambarus clarkii and Aquaculture Best Practices
by Peipei Wang, Jackson Samwel Bakari, Yanqiu Han, Honghui Hu, Zhilong Liu, Yewei Zhang, Zigui Chen, Chungui Huang, Miaomiao Wang, Huangen Chen, Xiaojun Jing and Shengyan Su
Animals 2026, 16(3), 495; https://doi.org/10.3390/ani16030495 - 5 Feb 2026
Viewed by 1440
Abstract
Temperature is a critical abiotic factor regulating the physiology, growth, and reproduction of ectothermic aquatic animals. In China, the rapid expansion of the red swamp crayfish Procambarus clarkii) industry faces significant challenges due to seasonal temperature fluctuations (optimal growth at 20–25 °C [...] Read more.
Temperature is a critical abiotic factor regulating the physiology, growth, and reproduction of ectothermic aquatic animals. In China, the rapid expansion of the red swamp crayfish Procambarus clarkii) industry faces significant challenges due to seasonal temperature fluctuations (optimal growth at 20–25 °C and reproduction favored at 18–22 °C). This review focuses specifically on TRP channels, particularly TRPA1 as a key thermosensor in crayfish, and on downstream signaling pathways involving heat shock proteins (HSPs) and antioxidant defenses. We further link these biological mechanisms to aquaculture applications by evaluating best management practices for mitigating thermal stress, including integrated rice–crayfish co-culture, recirculating aquaculture systems (RAS), molecular marker-assisted breeding for thermal tolerance, and nutritional modulation (e.g., probiotics and immunostimulants). By maintaining thermal stability within the optimal range and directly enhancing physiological resilience through genetic and nutritional interventions, these practices provide a foundation for more sustainable and climate-resilient crayfish aquaculture. Full article
(This article belongs to the Section Aquatic Animals)
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