{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T05:00:09Z","timestamp":1781586009207,"version":"3.54.5"},"reference-count":25,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T00:00:00Z","timestamp":1771977600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Open Recruitment for Key Scientific and Technological Projects","award":["2023ZXJ02B03"],"award-info":[{"award-number":["2023ZXJ02B03"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>This study aimed to scientifically assess cold stress in dairy calves and optimize winter rearing protocols. A combined approach of feeding trials, expert surveys, and multidimensional data analysis was used to evaluate the effects of outdoor (\u22125~\u221228 \u00b0C) and indoor (5 \u00b0C) environments on Holstein dairy calves. A 60-day controlled trial was conducted with 20 healthy 5-day-old calves. In parallel, an interdisciplinary panel of 20 experts and 8 farmers established a cold stress evaluation system via the analytic hierarchy process (AHP), with cold stress levels quantified through fuzzy comprehensive evaluation. Environmental (weight = 0.62), physiological (weight = 0.22), and behavioral (weight = 0.16) factors contributed differentially to cold stress assessment, with data showing that outdoor calves were under mild cold stress (maximum membership degree = 0.64). The temperature\u2013humidity index (THI) showed significant correlations with multiple physiological and biochemical parameters. Generalized linear mixed model (GLMM) analysis confirmed that THI variation significantly influenced calf standing time, respiratory rate (RR), malondialdehyde (MDA) content, and total antioxidant capacity (T-AOC). In feeding trials, indoor calves exhibited marginally higher average daily gain and body weight in early stages, whereas outdoor calves demonstrated significantly better growth performance by day 60. The outdoor group displayed increased lying and defecation behaviors, along with reduced locomotor\/standing time and respiratory frequency. No significant intergroup differences were observed in serum immune or antioxidant indicators. Metabolomic analysis identified 20 differentially expressed metabolites, indicating an enhancement in the activity of energy metabolism pathways in calves. This study establishes a quantitative methodology for cold stress evaluation, clarifies environment\u2013physiology\u2013behavior interactions, and provides a theoretical basis for winter calf management. The results confirm that outdoor cold exposure did not hinder calf growth without compromising health, offering scientific support for optimizing outdoor rearing strategies in cold regions.<\/jats:p>","DOI":"10.3390\/ani16050716","type":"journal-article","created":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T13:37:37Z","timestamp":1772026657000},"page":"716","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Analysis of Behavioral, Growth and Metabolic Indicators in Suckling Calves Under Outdoor Winter Rearing Conditions Using Fuzzy Comprehensive Evaluation"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7950-8875","authenticated-orcid":false,"given":"Jiachen","family":"Qu","sequence":"first","affiliation":[{"name":"Department of Animal Science, Nanjing Agricultural University, Nanjing 210018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1124-5831","authenticated-orcid":false,"given":"Xiaojing","family":"Zhou","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Green and Low-Carbon Agriculture in the Northeast Plain, Department of Information and Computing Science, Heilongjiang Bayi Agricultural University, Ministry of Agriculture and Rural Affairs, No. 5 Xinyang Road, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jintao","family":"Liu","sequence":"additional","affiliation":[{"name":"Lindian Youran Animal Husbandry Co., Ltd., Daqing 166300, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaoyu","family":"Han","sequence":"additional","affiliation":[{"name":"Department of Animal Science, Nanjing Agricultural University, Nanjing 210018, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yongli","family":"Qu","sequence":"additional","affiliation":[{"name":"The Key Laboratory of Green and Low-Carbon Agriculture in the Northeast Plain, Department of Information and Computing Science, Heilongjiang Bayi Agricultural University, Ministry of Agriculture and Rural Affairs, No. 5 Xinyang Road, Daqing 163319, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,2,25]]},"reference":[{"key":"ref_1","unstructured":"Sun, L. (2020). Effect of Cold Stress on Intestinal Microbiota and Body Metabolism in Piglets, South China Agricultural University."},{"key":"ref_2","unstructured":"Su, Y.Y. (2021). Influence of Long Term Cold Stimulation on the Environment Adaprability of Broiler Chickens, Northeast Agricultural University. Available online: https:\/\/kns.cnki.net\/kcms2\/article\/abstract?v=hmNf6PfbaBtLQmGmy6mGMif2UEkfhm5h3jYWXhgUWtqWoez3uTFQHXb3HI2CMMfYBMUp8I9ULohOUWoipBkIbl9ewxLd6CUU8GZ3XYoRVKydVJCY0ZrXDXGK2PBCY9v9Z5H1_xi7wBWTC0f4v1M0wdiLnwxtaf35PWe-4Vk-CJ2Ufbpr7AhnenoVPJxApOf5&uniplatform=NZKPT&language=CHS."},{"key":"ref_3","unstructured":"Gao, J.W. (2020). Effects of Severe Cold Stress on the mRNA Expression Levels of Lipid Metabolism-Related Genes and Fat Deposition in Altay and Hybrid Lambs, Shihezi University."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Mortazavi, M.S., Hajmohammadi, M., Buonaiuto, G., Colleluori, R., Lamanna, M., Cavallini, D., Valizadeh, R., Ebrahimi, S.H., and Oliveira, C.A.F. (2025). The Effect of a Pre-Mix of Essential Organic Minerals on Growth, Antioxidant Indices, and the Diarrhea Incidence in Dairy Calves Breed in Arid Climates. Ruminants, 5.","DOI":"10.3390\/ruminants5020022"},{"key":"ref_5","unstructured":"Bai, D.D. (2017). Effects of Different Seasons on Blood Biochemical Indexes and Expression of Genes Related to Cold and Heat Stress in Sanhe Cattle, Agricultural University of Inner Mongolia. Available online: https:\/\/kns.cnki.net\/kcms2\/article\/abstract?v=HHZEz91P5iI_sTQKhmROkzdm2eQ_YAqOGNGwU6sf6E7w97pguZoeAW2ZW_9BM5wMlsgNcwHJU_YiD0kEa6YB3damSoynylsGRtIoPQlxWI3Ihih8uAm743q2vC91vDLDjpaQKuthzJYwWbM-xmvYdbw-xb5t4alHJ0_0-826QDWTdAw34jv-og==&uniplatform=NZKPT&language=CHS."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S0749-0720(15)30590-9","article-title":"Cold Thermoregulation in the Newborn Calf","volume":"10","author":"Olson","year":"1994","journal-title":"Vet. Clin. North Am. Food Anim. Pract."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1007\/s11250-025-04363-1","article-title":"Impact of urea-molasses treated wheat straw levels in total mixed rations on growth and nutrient digestibility in Azikheli buffalo calves","volume":"57","author":"Rashid","year":"2025","journal-title":"Trop. Anim. Health Prod."},{"key":"ref_8","first-page":"54","article-title":"Cold or heat stress determination of barn cows on temperature and humidity index in Hohhot","volume":"36","author":"Xu","year":"2005","journal-title":"J. Domest. Anim. Ecol."},{"key":"ref_9","unstructured":"National Academies of Sciences, Engineering, and Medicine (NASEM) (2021). Nutrient Requirements of Dairy Cattle, National Academies Press."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"9137","DOI":"10.3168\/jds.2022-21828","article-title":"Research and application of a new multilevel fuzzy comprehensive evaluation method for cold stress in dairy cows","volume":"105","author":"Fu","year":"2022","journal-title":"J. Dairy Sci."},{"key":"ref_11","unstructured":"Jing, X. (2006). Study on Effects of Ecological Chronic Cold and Heat Stress on Maintenance Behavior and Immune Function of Holstein Dairy Cows, Inner Mongolia Agricultural University. Available online: https:\/\/kns.cnki.net\/kcms2\/article\/abstract?v=HHZEz91P5iJMbK8OgwuiJScguwlGHMBGWtArmQ617S4X23xxkS4LpM_e9xPc9kKXh-0D5jEpXg40yHIxUlMWGCnqlQV-NBisrUhUZBjutS62BS1Eg85Y0tmnbxpIEXe53RjBSjBLTl2WPJsYEsRpQOQmWxnhCZ1rjEAiEjK0LU2oRrtneqvrSU5Yu4GqMSIO&uniplatform=NZKPT&language=CHS."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"898","DOI":"10.3382\/ps.2013-03438","article-title":"Effects of low ambient temperatures and dietary vitamin C supplement on growth performance, blood parameters, and antioxidant capacity of 21-day-old broilers","volume":"93","author":"Yang","year":"2014","journal-title":"Poult. Sci."},{"key":"ref_13","first-page":"29","article-title":"Effects of different feeding patterns on digestion, metabolism and Growth Performance of pre-weaning calves","volume":"13","author":"Ren","year":"2012","journal-title":"Chin. Dairy Cows"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2438","DOI":"10.3168\/jds.2015-9901","article-title":"Invited review: Influence of climatic conditions on development, performance, and health of calves","volume":"99","author":"Roland","year":"2016","journal-title":"J. Dairy Sci."},{"key":"ref_15","first-page":"22","article-title":"Effect of chronic hot and cold stress on serum indexes of breeding cows","volume":"1","author":"Shi","year":"2023","journal-title":"China Feed"},{"key":"ref_16","unstructured":"Zhou, G.C. (2019). Effect of Wind Speed on Physiological Indexes, Antioxidant and Immune Levels of Sheep Under Low Temperature Conditions, Northwest A&F University. Available online: https:\/\/kns.cnki.net\/kcms2\/article\/abstract?v=hmNf6PfbaBvrtjRvC9_AYrye0unAPtAMNf0doU_LTmKUUq9vewdYlhQqpnOV6cfkp5xTEE3gPSmytil2lowUAYGKyZMIzG63NMPIqAxWLY1EZ2vVcffvfVXkeG7gH-tC9xbAEKkt2TKZRagUwNzcpt0m9uuqAaUsy-jePLjDEg45IYqsVjhtUg==&uniplatform=NZKPT&language=CHS."},{"key":"ref_17","first-page":"60","article-title":"Effects of temperature and humidity index on production performance and physiological and biochemical indexes of dairy cows in different seasons","volume":"45","author":"Wang","year":"2009","journal-title":"Chin. J. Anim. Husb."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1360","DOI":"10.1016\/j.cell.2015.11.004","article-title":"Gut Microbiota Orchestrates Energy Homeostasis during Cold","volume":"163","author":"Chevalier","year":"2015","journal-title":"Cell"},{"key":"ref_19","first-page":"16","article-title":"Research on physiological indices and disease resistance of Freiherr and Holstein F1cattle and Holstein cows under high temperature and cold conditions","volume":"8","author":"Lin","year":"2023","journal-title":"China Dairy Cattle"},{"key":"ref_20","first-page":"65","article-title":"Influence of chronic cold stress on immune-related indexes of Simmental crossbred calves","volume":"15","author":"Meng","year":"2010","journal-title":"J. China Agric. Univ."},{"key":"ref_21","unstructured":"Yao, R. (2021). Influence of Temperature-Humidity Index (THI) on Growth Performance, Physiological Parameters and Blood Indexes of Weaned Lambs, Hebei Agricultural University."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1038\/nature02488","article-title":"Citric acid cycle intermediates as ligands for orphan G-protein-coupled receptors","volume":"429","author":"He","year":"2004","journal-title":"Nature"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1111\/j.1365-2672.2004.02282.x","article-title":"PCR detection and 16S rRNA sequence-based phylogeny of a novel Propionibacterium acidipropionici applicable for enhanced fermentation of high moisture corn","volume":"97","author":"Romanov","year":"2004","journal-title":"J. Appl. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"42","DOI":"10.2527\/jas.2005-651","article-title":"Genetic parameters for body weight, hip height, and the ratio of weight to hip height from random regression analyses of Brahman feedlot cattle","volume":"85","author":"Riley","year":"2007","journal-title":"J. Anim. Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"4578","DOI":"10.1073\/pnas.1000081107","article-title":"Succession of microbial consortia in the developing infant gut microbiome","volume":"108","author":"Spor","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/5\/716\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,25]],"date-time":"2026-02-25T13:44:40Z","timestamp":1772027080000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/5\/716"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,2,25]]},"references-count":25,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,3]]}},"alternative-id":["ani16050716"],"URL":"https:\/\/doi.org\/10.3390\/ani16050716","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,2,25]]}}}