Animals最新文献

{"title":"Dolphin Pituitary Gland: Immunohistochemistry and Ultrastructural Cell Characterization Following a Novel Anatomical Dissection Protocol and Non-Invasive Imaging (MRI).","authors":"Paula Alonso-Almorox, Alfonso Blanco, Carla Fiorito, Eva Sierra, Cristian Suárez-Santana, Francesco Consolli, Manuel Arbelo, Raiden Grandía Guzmán, Ignacio Molpeceres-Diego, Antonio Fernández Gómez, Javier Almunia, Ayoze Castro-Alonso, Antonio Fernández","doi":"10.3390/ani15050735","DOIUrl":"10.3390/ani15050735","url":null,"abstract":"<p><p>The pituitary gland regulates essential physiological processes in mammals. Despite its importance, research on its anatomy and ultrastructure in dolphins remains scarce. Using non-invasive imaging technology (MRI) and a novel skull-opening and dissection protocol, this study characterizes the dolphin pituitary through immunohistochemistry (IHC) and transmission electron microscopy (TEM). A total of 47 pituitaries were collected from stranded common bottlenose dolphins (<i>Tursiops truncatus</i>). common dolphins (<i>Delphinus delphis</i>), and Atlantic spotted dolphins (<i>Stenella frontalis</i>). as well as from captive common bottlenose dolphins. MRI allowed visualization of the gland's anatomy and its spatial relationship with the hypothalamus and surrounding structures. A modified skull-opening and pituitary extraction protocol ensured the preservation of the adenohypophysis and neurohypophysis for detailed analysis. Histological, immunohistochemical, and ultrastructural studies confirmed the gland's structural organization, identifying eight distinct adenohypophyseal cell types: corticotrophs (ACTH), somatotrophs (GH), gonadotrophs (FSH and LH), lactotrophs (LTH), melanotrophs (MSH), thyrotrophs (TSH), follicular cells, and capsular cells. This study presents the first immunolabelling of thyrotrophs in cetacean adenohypophysis and the first detailed ultrastructural characterization of adenohypophyseal cells in cetaceans, providing baseline data for future research. By integrating multidisciplinary techniques, it advances the understanding of dolphin neuroendocrinology and highlights broader implications for cetacean health, welfare, and conservation.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11898967/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A New Species of the Genus <i>Acrossocheilus</i> Oshima, 1919 (Cypriniformes: Cyprinidae) from the Dabie Mountains.","authors":"Tian-En Chen, Jia-Xin Xu, Peng-Ju Li, Huan-Fu Hu, Kai Gao, Hai-Peng Zhao","doi":"10.3390/ani15050734","DOIUrl":"10.3390/ani15050734","url":null,"abstract":"<p><p><i>Acrossocheilus</i> Oshima, 1919, a cyprinid genus of Cyprinidae in southern China and currently comprises 26 valid species. In this study, we describe <i>Acrossocheilus dabieensis</i><b>sp. nov.</b> from the Dabie Mountains, China. This new species differs from its congeners by the following combination of characters: The second primary vertical bar (PB2) is situated beneath the anterior origin of the dorsal fin in females and subadult males. Vertical bars extend to the end of the ventral abdomen in juveniles, and they gradually recede above the lateral line in adult females, whereas they are absent in adult males. The last unbranched dorsal-fin ray is slender with a smooth posterior margin. Phylogenetic analyses based on the mitochondrial DNA sequences indicate that <i>A. dabieensis</i><b>sp. nov.</b> is a monophyletic group, and it forms a sister group with <i>A. kreyenbergii</i>, reinforcing the status of the new species. A key to the barred species of <i>Acrossocheilus</i> is also provided.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899002/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612959","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study on the Regulatory Mechanisms of Carapace Marking Formation in <i>Marsupenaeus japonicus</i>.","authors":"Panpan Wang, Jiawei Zhu, Huanyu Chen, Qingyuan Hu, Zhenxiang Chen, Wenjia Li, Ting Yang, Jin Zhu, Binlun Yan, Huan Gao, Chaofan Xing","doi":"10.3390/ani15050727","DOIUrl":"10.3390/ani15050727","url":null,"abstract":"<p><p>There are two phenotypes in the natural populations of <i>Marsupenaeus japonicus</i>, which is an ideal model for studying the formation of markings and body color in crustaceans. In a previous study, we used comparative transcriptome technology to screen some functional genes related to body color regulation. Here, high-throughput sequencing technology was used to perform microRNA (miRNA) sequencing analysis on the exoskeleton of <i>M. japonicus</i> with two types of carapace markings, and functional studies of related genes were performed. A total of 687 mature miRNAs belonging to 135 miRNA families were identified in this study, and 111 novel miRNAs were found. Through stringent screening conditions, a total of 18 differentially expressed miRNAs were identified, including 14 with upregulated expression and 4 with downregulated expression. Multiple target genes were predicted for almost all of the differentially expressed miRNAs. The expression levels of several target genes, such as those related to cytoplasmic microtubule organization, transmembrane transportation, and signal transduction, were confirmed using qRT-PCR. This study revealed that both the <i>CRCN A2</i> and <i>CRCN C1</i> genes were highly expressed in type I individuals, while the expression levels of their related miRNAs in type I individuals were lower than those in type II individuals, which is consistent with the mechanism of miRNAs negatively regulating mRNA expression. Through interference with the <i>CRCN A2</i> and <i>CRCN C1</i> genes, a clear regulatory relationship was found between the two genes, and the dendritic xanthophores in the carapace of <i>M. japonicus</i> gradually changed from bright yellow to dark black, with obvious shrinkage. In summary, our studies provide references for the regulatory mechanisms of marking formation in <i>M. japonicus</i>.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899420/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Why Are Some Snakes More Terrifying and What Is Behind the Fear?","authors":"Daniel Frynta, Markéta Janovcová, Hassan Sh Abdirahman Elmi, Iveta Štolhoferová, Veronika Rudolfová, Kateřina Rexová, David Sommer, David Král, Daniel Alex Berti, Eva Landová, Petra Frýdlová","doi":"10.3390/ani15050731","DOIUrl":"10.3390/ani15050731","url":null,"abstract":"<p><p>Snakes are stimuli inducing an ancestral fear response in humans and other primates. Certain snakes evoke more subjective fear than others. True vipers are high-fear-eliciting snakes for both African and European respondents. This can be explained by the evolutionary experience of human ancestors in Africa. The question arises as to how snakes living in the Americas and Australia, with which humans have no evolutionary experience, will be evaluated. While these snakes belong to broader taxonomic groups that have distant relatives in the Old World, they have evolved independently for tens of millions of years. We prepared a set of 32 pictures depicting eight American pit vipers, eight Australian elapids, eight constrictors, and eight colubrids and asked the respondents to rank these stimuli according to the fear these snakes evoke. Here, we show a high cross-cultural agreement between evaluations by African and European respondents. Snakes characterized by a robust body shape, such as American pit vipers, Australian death adders, pythons, and boas, were the most fear-evoking. The body width was the strongest predictor of evoked fear. The contribution of coloration and pattern of the stimulus to the fear response was not proved. This supports the view that the patterns of fear are not dependent on direct experience, but its underlying mechanisms are shared cross-culturally.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11898634/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613159","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insight into the Potential of Somatostatin Vaccination with Goats as a Model: From a Perspective of the Gastrointestinal Microbiota.","authors":"Xiaoli Zhang, Juncai Chen, Siqi Zhang, Bingni Wei, Yanguo Han, Zhongquan Zhao","doi":"10.3390/ani15050728","DOIUrl":"10.3390/ani15050728","url":null,"abstract":"<p><p>Deciphering the gastrointestinal microbial response to oral SS DNA vaccines with different doses is helpful for identifying the mechanism for effective utilization of the vaccine for improving animal production. Here, we conduct a comparative study with different doses of vaccine (control: empty plasmid; low dose: 1 × 10⁷ CFU vaccine; high dose: 1 × 10¹² CFU vaccine) using goat as a case to investigate the potential of somatostatin vaccination from the entire gastrointestinal microbiota perspective. Our results show that body weight gain and slaughter rate are greater in the L_SS group than in the C_SS group. Compared with the C_SS group, the GH concentration is reduced, while the SS concentration is elevated in the cecum of L_SS goats. Moreover, the SCFAs concentration is elevated in the L_SS goats, the acetate molar proportion is lower in the rumen, the proportion of the acetate is decreased, and propionate is increased in the cecum of L_SS goats. Our data indicate that the low-dose somatostatin vaccine possesses a more efficient improvement in the productivity of goats, emphasizing that the dosage should be considered to reach its optimal effect on the host. Moreover, we find that different doses of the SS vaccination select distinct microbial communities in the gastrointestinal tract. Beta diversity analysis shows a significant interaction. Microorganisms capable of converting nutrients, including <i>Ruminococcacease</i>, <i>Butyrivibrio</i>, <i>Akkermansia</i>, and <i>Lachnospiraceae</i> are enriched, altering the gastrointestinal fermentation response to SS DNA vaccination of ruminants. Moreover, the correlation analysis results revealing these biomarkers have a close association with the phenotypes of productivity. These results imply that somatostatin immunoneutralization might directly alter the gastrointestinal tract commensal bacterial structure, improving gastrointestinal homeostasis, and, thus, modifying the fermentability and effected hormone level to improve the productivity of goats. Our study extends the understanding of the somatostatin vaccine regulation of ruminants' growth through the entire gastrointestinal microbial perspective.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899232/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613116","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Meningeal Lymphatic and Glymphatic Structures in a Pelagic Delphinid (<i>Delphinus delphis</i>).","authors":"Olivia N Jackson, Tiffany F Keenan, Nathan P Nelson-Maney, Sentiel A Rommel, William A McLellan, D Ann Pabst, Alexander M Costidis, Kathleen M Caron, Dawn N Kernagis, David S Rotstein, Victoria G Thayer, Craig A Harms, Marina A Piscitelli-Doshkov, Paul Doshkov, Lorian E Schweikert, Kara E Yopak, Molly Braun, Michael S Tift","doi":"10.3390/ani15050729","DOIUrl":"10.3390/ani15050729","url":null,"abstract":"<p><p>The glymphatic system, an analog of the peripheral lymphatic system in the brain, and the meningeal lymphatic system are critical to central nervous system health. The glymphatic system functions to distribute cerebrospinal fluid and important compounds throughout the brain and to remove metabolic waste. The flow of cerebrospinal fluid through this system is affected by changes in cerebral blood flow, intracranial pressure, and vascular tone. Cetaceans experience profound cardiorespiratory alterations while diving that can directly affect cerebrospinal fluid and blood flow and, thus, glymphatic function. Our goal was to investigate glymphatic and lymphatic system structures, including perivascular spaces, aquaporin-4 water channels, meningeal lymphatic, and dural venous sinus vessels in the common dolphin (<i>Delphinus delphis</i>), using immunofluorescent labeling, histochemical staining, and postmortem computed tomography (CT) angiography. We highlight perivascular spaces and aquaporin-4 water channels surrounding blood vessels in the parenchyma and demonstrate evidence of meningeal lymphatic vessels and associated dural venous sinuses. These results demonstrate that common dolphins possess the key anatomical structures required for functional glymphatic and meningeal lymphatic systems. Future studies can build upon these anatomical discoveries to study the function and role of these systems in brain health in this species.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899484/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613139","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of Fermented Liquid Feed with Compound Probiotics on Growth Performance, Meat Quality, and Fecal Microbiota of Growing Pigs.","authors":"Mengting Ji, Xiaoyin Rong, Yifan Wu, Haonan Li, Xiaolei Zhao, Yan Zhao, Xiaohong Guo, Guoqing Cao, Yang Yang, Bugao Li","doi":"10.3390/ani15050733","DOIUrl":"10.3390/ani15050733","url":null,"abstract":"<p><p>Feed fermentation enhances feed nutrition and animal health, but its impact on the gut microbiota of young pigs remains unclear. This study aimed to evaluate the effects of a probiotic fermented feed, which includes <i>Lactobacillus plantarum</i>, <i>Pediococcus pentosaceus</i>, <i>Bacillus subtilis</i>, and <i>Bacillus coagulans</i>, on the growth performance, meat quality, and intestinal microbiota of growing pigs. We randomly assigned 24 Duroc × Landrace × Landrace pigs to two groups: a control (Ctrl) group and the fermented liquid feed (FLF) group, with three replicate pens per group and four pigs per pen. Results indicated that the FLF group experienced a significant decrease in anti-nutritional factors like α-conglycinin and β-conglycinin. In addition, the average daily gain of pigs in the FLF group increased significantly, while the feed conversion ratio and shear force decreased. HE staining showed that the FLF group had notably enhanced villus height in the jejunum and ileum. 16S rRNA sequencing revealed a marked increase in the relative abundance of certain microbes in the FLF group, which were predominantly associated with carbohydrate and amino acid metabolism. These results indicated that compound probiotic FLF can elevate feed quality, enhance the growth performance of growing pigs, and ameliorate the structure of the gut microbiota.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899608/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Use of Cognition by Amphibians Confronting Environmental Change: Examples from the Behavioral Ecology of Crawfish Frogs (<i>Rana areolata</i>).","authors":"Michael J Lannoo, Rochelle M Stiles","doi":"10.3390/ani15050736","DOIUrl":"10.3390/ani15050736","url":null,"abstract":"<p><p>Amphibian conservation concerns frequently center on the idea of 'saving' them, with the underlying assumption they are the passive victims of anthropogenic environmental change. But this approach ignores the physiological, biochemical, and behavioral flexibility amphibians have employed since they first evolved ~365 million years ago. One overlooked advantage amphibians possess in the struggle for survival, and one humans might use in their efforts to conserve them, is their brains share the same blueprint as human brains, which allows them to acquire knowledge and understanding through experiences-in other words, amphibians have cognitive capabilities that assist them in their effort to survive. Here, we use four examples from our work on the behavioral ecology of Crawfish Frogs (<i>Rana areolata</i>) to form hypotheses about how cognition affects amphibian reaction to environmental and social change. The first two examples describe Crawfish Frog responses to seasonality and reproductive status, the third details their reaction to ecological disturbance, and the fourth describes how their response to the same stimulus changes with growth/age. In each example, we detail the neuronal circuitry thought to be involved and hypothesize the role of cognition. We propose that as one component of our fight to conserve amphibians, researchers should consider the full range of anatomical, physiological, biochemical, and behavioral features amphibians themselves employ in their defense, which are features responsible for their historical evolutionary success up until the Anthropocene. Further, we submit that acknowledging amphibians possess cognitive abilities can enrich interpretations of not only behavioral and ecological observations but also of neuroanatomical and neurophysiological results.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11898707/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143613171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Dispersal Limitation Dominates Riverine Fish Communities in the Areas of the Water Diversion Project in the Western Sichuan Plateau, China.","authors":"Tao Chang, Zheng Gong, Kunyu Shang, Piao Hu","doi":"10.3390/ani15050730","DOIUrl":"10.3390/ani15050730","url":null,"abstract":"<p><p>The riverine fish species are highly vulnerable and responsive to large-scale water diversion projects. These adverse impacts are more pronounced in the plateau river ecosystems, which may change the environmental conditions of fish habitats and community structure. We investigated the effects of various environmental factors on fish diversity in seven rivers of the Western Sichuan Plateau, which is the planned area of China's South-to-North Water Diversion Project. Twenty-two fish species, including eight exotic species, were collected during September 2023 (Autumn) and May 2024 (Spring). The fish communities exhibited no significant difference between seasons but had prominent variations among different rivers. The heterogeneity of fish communities was significantly and positively correlated with the geographical distance between the sampling sites (based on a projected coordinate system). Furthermore, the canonical correspondence analysis (CCA) illustrated that altitude contributed more to the distribution of fish species than other physicochemical factors, such as channel width, conductivity, and water temperature. Rivers at low altitudes are likely to be vulnerable to invasion of exotic fish. Our results demonstrated that the dispersal limitation by geographical distance and altitudinal gradient were the primary regulatory factors on the spatial differentiation of fish communities in the rivers of the study area, which reflected a high dependence of fish species on local habitats. As the water diversion project is implemented, more attention is expected to be paid to protecting fish habitats and regime shifts in fish communities. Additionally, the risk assessment of biological invasion under inter-basin water transfers and human activities should be carried out as soon as possible.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11898623/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612970","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Afternoon Calcium and Vitamin D Supplementation in Water: A Targeted Approach to Improve Laying Hen Nutrition.","authors":"Nasima Akter, Thi Hiep Dao, Tamsyn M Crowley, Sukirno, Aamir Nawab, Amy F Moss","doi":"10.3390/ani15050720","DOIUrl":"10.3390/ani15050720","url":null,"abstract":"<p><p>A modified AM/PM strategy was evaluated in this study by supplementing calcium (Ca) in afternoon/evening (PM) drinking water instead of feed, offering an alternative to supplying two diets. A 10-week trial was conducted with 288 Hy-Line Brown hens (56 weeks old) housed on the floor in 18 pens (16 hens per pen; 9 replicate pens per treatment). The control diet contained 4.5% Ca, while the AM/PM treatment had 4.1% in feed with additional Ca and vitamin D supplemented through PM drinking water, with other nutrients kept constant in both diets. Daily egg production and weekly feed and water consumption were recorded. Hen weight, welfare scores, and egg quality were assessed at weeks 5 and 10. Blood, ileal digesta, and tibia samples were collected (from 4 hens/pen) at week 10 for serum Ca and vitamin D, digestibility, and bone analyses. Liver and kidney weights, as well as keel bone scores, were also recorded at week 10. Hens on the AM/PM diet consumed significantly less water (171.23 vs. 196.85 mL/bird/day; <i>p</i> < 0.001) and had a significantly higher serum vitamin D (27.59 vs. 22.64 ng/mL; <i>p</i> = 0.014), with a tendency towards elevated serum calcium levels (31.99 vs. 29.74 mg/dL; <i>p</i> = 0.089) compared to the control group. No differences in laying performance, co-efficient of variance (CV%) in individual hen and egg weight, liver and kidney weight, or keel and tibia characteristics were observed between treatments (<i>p</i> > 0.05). Similarly, there was no treatment effect on egg quality, and welfare parameters at week 5 and 10, except for smaller eggs (57.13 vs. 57.96; <i>p</i> = 0.015), and reduced chest (0.96 vs. 1.16; <i>p</i> = 0.028) and belly feather damage (1.34 vs. 1.55; <i>p</i> = 0.021), respectively, in hens of the AM/PM treatment compared to the control at week 10. The AM/PM treatment improved ileal digestible energy as is (9.00 vs. 8.22; <i>p</i> = 0.022) and as dry matter (10.00 vs. 9.06; <i>p</i> = 0.014) compared to the control one. Overall, it demonstrated comparable performance to the control, with added benefits of reduced water intake, better feather condition and vitamin D level and energy digestibility.</p>","PeriodicalId":7955,"journal":{"name":"Animals","volume":"15 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11899453/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}