{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T06:46:40Z","timestamp":1783666000242,"version":"3.55.0"},"reference-count":43,"publisher":"Public Library of Science (PLoS)","issue":"10","license":[{"start":{"date-parts":[[2023,10,5]],"date-time":"2023-10-05T00:00:00Z","timestamp":1696464000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Higher Education Comission of Pakistan"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Objectives were to determine the effects of supplementing increasing amounts of choline ion on hepatic composition and mRNA abundance in pregnant dry cows subjected to a fatty liver induction protocol. Holstein cows (35 primiparous and 41 multiparous) at mean (\u00b1 standard deviation) of 211 \u00b1 9.9 days of gestation were blocked by body condition (3.59 \u00b1 0.33) and assigned to receive 0, 6.45, 12.90, 19.35, and 25.80 g\/day of choline ion as rumen-protected choline (RPC) as a top-dress for 14 days. Cows were fed for ad libitum intake on days 1 to 5 and restricted to 30% of the required net energy for lactation from days 6 to 14 of the experiment. Hepatic tissue was sampled on days 5 and 14 and analyzed for concentrations of triacylglycerol and glycogen, and mRNA abundance was investigated. Orthogonal contrasts evaluated the effects of supplementing RPC (0 g\/day vs. rest), and the linear, quadratic, and cubic effects of increasing intake of choline ion from 6.45 to 25.80 g\/day. Results are depicted in sequence of treatments from 0 to 25.8. During feed restriction, RPC reduced the concentration of hepatic triacylglycerol by 28.5% and increased that of glycogen by 26.1%, and the effect of increasing RPC intake on triacylglycerol was linear (6.67 vs. 5.45 vs. 4.68 vs. 5.13 vs. 3.81 \u00b1 0.92% wet-basis). Feeding RPC during feed restriction increased abundance of transcripts involved in choline metabolism (<jats:italic>CHKA<\/jats:italic>,<jats:italic>PLD1<\/jats:italic>), synthesis of apolipoprotein-B100 (<jats:italic>APOB100<\/jats:italic>), and antioxidant activity (<jats:italic>GPX3<\/jats:italic>), and decreased the abundance of transcripts involved in hepatic lipogenesis (<jats:italic>DGAT2<\/jats:italic>,<jats:italic>SREBF1<\/jats:italic>) and acute phase response (<jats:italic>SAA3<\/jats:italic>). Most effects were linear with amount of choline fed. Changes in hepatic mRNA abundance followed a pattern of reduced lipogenesis and enhanced lipids export, which help explain the reduced hepatic triacylglycerol content in cows fed RPC. Choline exerts lipotropic effects in dairy cows by altering transcript pathways linked to hepatic lipids metabolism.<\/jats:p>","DOI":"10.1371\/journal.pone.0290562","type":"journal-article","created":{"date-parts":[[2023,10,5]],"date-time":"2023-10-05T17:46:57Z","timestamp":1696528017000},"page":"e0290562","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Dose-dependent effects of rumen-protected choline on hepatic metabolism during induction of fatty liver in dry pregnant dairy cows"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5312-4523","authenticated-orcid":true,"given":"Usman","family":"Arshad","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcos G.","family":"Zenobi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paula","family":"Tribulo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charles R.","family":"Staples","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3403-1465","authenticated-orcid":true,"given":"Jos\u00e9 E. 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Dairy Sci"},{"key":"pone.0290562.ref003","first-page":"e814808","article-title":"Qualitative and quantitative changes in total lipid concentration and lipid fractions in liver tissue of periparturient German Holstein dairy cows of two age groups","volume":"9","author":"KB Theinert","year":"2022","journal-title":"PLoS One"},{"key":"pone.0290562.ref004","doi-asserted-by":"crossref","first-page":"5393","DOI":"10.3168\/jds.2021-21031","article-title":"Hepatic triacylglycerol associations with production and health in dairy cows","volume":"105","author":"U Arshad","year":"2022","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref005","doi-asserted-by":"crossref","first-page":"2772","DOI":"10.3168\/jds.S0022-0302(89)79421-2","article-title":"In vitro degradation of choline from selected feedstuffs and choline supplements","volume":"72","author":"BK Sharma","year":"1989","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref006","doi-asserted-by":"crossref","first-page":"2413","DOI":"10.3168\/jds.2006-028","article-title":"Supplemental choline for prevention and alleviation of fatty liver in dairy cattle","volume":"90","author":"RF Cooke","year":"2007","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref007","doi-asserted-by":"crossref","first-page":"5902","DOI":"10.3168\/jds.2017-13973","article-title":"Feeding increasing amounts of ruminally protected choline decreased fatty liver in nonlactating, pregnant Holstein cows in negative energy status","volume":"101","author":"MG Zenobi","year":"2018","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref008","doi-asserted-by":"crossref","first-page":"282","DOI":"10.3168\/jds.2019-16842","article-title":"Meta-analysis of the effects of supplemental rumen-protected choline during the transition period on performance and health of parous dairy cows","volume":"103","author":"U Arshad","year":"2020","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref009","doi-asserted-by":"crossref","first-page":"1102","DOI":"10.3168\/jds.2012-5396","article-title":"Effect of rumen-protected choline supplementation on liver and adipose gene expression during the transition period in dairy cattle","volume":"96","author":"RMA Goselink","year":"2013","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref010","doi-asserted-by":"crossref","first-page":"3864","DOI":"10.3168\/jds.S0022-0302(93)77728-0","article-title":"Lipid absorption and transport in ruminants","volume":"76","author":"D. Bauchart","year":"1993","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref011","doi-asserted-by":"crossref","first-page":"2998","DOI":"10.1016\/S0021-9258(18)69166-5","article-title":"The active synthesis of phosphatidylcholine is required for very low-density lipoprotein secretion from rat hepatocytes","volume":"263","author":"ZM Yao","year":"1988","journal-title":"J. Biol. Chem"},{"issue":"8","key":"pone.0290562.ref012","doi-asserted-by":"crossref","first-page":"e103412","DOI":"10.1371\/journal.pone.0103412","article-title":"Choline and choline metabolite patterns and associations in blood and milk during lactation in dairy cows","volume":"9","author":"VM Artegoitia","year":"2014","journal-title":"PLoS One"},{"key":"pone.0290562.ref013","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1186\/1746-6148-10-122","article-title":"Metabolomic biomarkers correlating with hepatic lipidosis in dairy cows","volume":"10","author":"S Imhasly","year":"2014","journal-title":"BMC Vet. Res"},{"key":"pone.0290562.ref014","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.anireprosci.2013.07.014","article-title":"Lipid mobilization and oxidative stress as metabolic adaptation processes in dairy heifers during transition period","volume":"141","author":"R Turk","year":"2013","journal-title":"Anim. Reprod. Sci"},{"key":"pone.0290562.ref015","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1292\/jvms.64.293","article-title":"Relevance of apolipoproteins in the development of fatty liver and fatty liver- related peripartum diseases in dairy cows","volume":"64","author":"N. Katoh","year":"2002","journal-title":"J. Vet. Med. Sci"},{"key":"pone.0290562.ref016","doi-asserted-by":"crossref","first-page":"1451","DOI":"10.3945\/jn.109.108233","article-title":"Daily injection of tumor necrosis factor-\u03b1 increases hepatic triglycerides and alters transcript abundance of metabolic genes in lactating dairy cattle","volume":"139","author":"BJ Bradford","year":"2009","journal-title":"J. Nutr"},{"issue":"282","key":"pone.0290562.ref017","doi-asserted-by":"crossref","first-page":"27155","DOI":"10.1074\/jbc.M704408200","article-title":"Anti-inflammatory effects of phosphatidylcholine","volume":"14","author":"I Treede","year":"2007","journal-title":"J. Biol. Chem"},{"key":"pone.0290562.ref018","volume-title":"Nutrient Requirements of Dairy Cattle.","author":"NRC","year":"2001","edition":"7"},{"key":"pone.0290562.ref019","volume-title":"Nutrient Requirements of Dairy Cattle","author":"NASEM (National Academies of Sciences, Engineering, and Medicine)","year":"2021","edition":"8"},{"key":"pone.0290562.ref020","doi-asserted-by":"crossref","first-page":"146","DOI":"10.1016\/j.ygeno.2009.04.008","article-title":"A powerful and flexible linear mixed model framework for the analysis of relative quantification RT-PCR data","volume":"94","author":"JP Steibel","year":"2009","journal-title":"Genomics"},{"key":"pone.0290562.ref021","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1186\/1471-2105-7-85","article-title":"Statistical analysis of real-time PCR data","volume":"7","author":"JS Yuan","year":"2006","journal-title":"BMC Bioinformatics"},{"key":"pone.0290562.ref022","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1093\/nar\/gkw419","article-title":"Heatmapper: Web-enabled heat mapping for all","volume":"44","author":"S Babicki","year":"2016","journal-title":"Nucleic Acids Res"},{"key":"pone.0290562.ref023","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1111\/j.2517-6161.1964.tb00553.x","article-title":"An analysis of transformations","volume":"26","author":"G Box","year":"1964","journal-title":"J. R. Stat. Soc. Series B Stat. Methodol"},{"key":"pone.0290562.ref024","doi-asserted-by":"crossref","first-page":"865","DOI":"10.2134\/agronj2008.0226x","article-title":"Data transformation in statistical analysis of field trials with changing treatment variance","volume":"101","author":"HP Piepho","year":"2009","journal-title":"Agron. J"},{"key":"pone.0290562.ref025","unstructured":"J\u00f8rgensen E, Pedersen AR. How to obtain those nasty standard errors from transformed data\u2013and why they should not be used. Page 20 in Biometry Research Unit\u2014Internal Report 7. 1998. Danish Institute of Agricultural Sciences."},{"key":"pone.0290562.ref026","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/S0021-9258(19)50785-2","article-title":"The function of cytidine coenzymes in the biosynthesis of phospholipides","volume":"222","author":"EP Kennedy","year":"1956","journal-title":"J. Biol. Chem"},{"key":"pone.0290562.ref027","doi-asserted-by":"crossref","first-page":"9732","DOI":"10.3168\/jds.2016-11382","article-title":"Choline absorption and evaluation of bioavailability markers when supplementing choline to lactating dairy cows","volume":"99","author":"MJ de Veth","year":"2016","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref028","doi-asserted-by":"crossref","first-page":"1813","DOI":"10.3168\/jds.S0022-0302(88)79750-7","article-title":"Triglyceride accumulation and very low-density lipoprotein secretion by rat and goat hepatocytes in vitro","volume":"71","author":"BB Kleppe","year":"1988","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref029","doi-asserted-by":"crossref","first-page":"338","DOI":"10.1016\/S1050-1738(01)00071-8","article-title":"Intracellular assembly of VLDL: two major steps in separate cell compartments","volume":"10","author":"SO Olofsson","year":"2000","journal-title":"Trends Cardio. Med"},{"key":"pone.0290562.ref030","doi-asserted-by":"crossref","first-page":"833","DOI":"10.3168\/jds.2018-15208","article-title":"Perilipin 5 promotes hepatic steatosis in dairy cows through increasing lipid synthesis and decreasing very low-density lipoprotein assembly","volume":"102","author":"H Jia","year":"2019","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref031","doi-asserted-by":"crossref","first-page":"e0171080","DOI":"10.1371\/journal.pone.0171080","article-title":"Choline and methionine differentially alter methyl carbon metabolism in bovine neonatal hepatocytes","volume":"12","author":"TL Chandler","year":"2017","journal-title":"PLoS One"},{"key":"pone.0290562.ref032","first-page":"106","article-title":"Effect of source and amount of rumen-protected choline on hepatic metabolism during induction of fatty liver in dairy cows","author":"U Arshad","year":"2023","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref033","first-page":"106","article-title":"Rumen-protected choline reduces hepatic lipidosis by increasing hepatic triacylglycerol-rich lipoprotein secretion in dairy cows","author":"U Arshad","year":"2023","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref034","doi-asserted-by":"crossref","first-page":"e0160659","DOI":"10.1371\/journal.pone.0160659","article-title":"Regulation of nutritional metabolism in transition dairy cows: Energy homeostasis and health in response to post-ruminal choline and methionine","volume":"11","author":"F Sun","year":"2016","journal-title":"PLoS One"},{"key":"pone.0290562.ref035","doi-asserted-by":"crossref","first-page":"3958","DOI":"10.3168\/jds.2016-11812","article-title":"Supplementation with rumen-protected methionine or choline during the transition period influences whole-blood immune response in periparturient dairy cows","volume":"100","author":"M Vailati-Riboni","year":"2017","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref036","doi-asserted-by":"crossref","first-page":"2200","DOI":"10.3168\/jds.2019-17378","article-title":"Effect of prepartum energy intake and supplementation with ruminally protected choline on innate and adaptive immunity of multiparous Holstein cows","volume":"103","author":"MG Zenobi","year":"2020","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref037","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1111\/j.1530-0277.1997.tb04250.x","article-title":"Choline deficiency augments and antibody to tumor necrosis factor attenuates endotoxin-induced hepatic injury","volume":"21","author":"CE Eastin","year":"1997","journal-title":"Alcohol. Clin. Exp. Res"},{"key":"pone.0290562.ref038","doi-asserted-by":"crossref","first-page":"9896","DOI":"10.3168\/jds.2021-21689","article-title":"Effects of maternal choline supplementation on performance and immunity of progeny from birth to weaning","volume":"105","author":"MG Zenobi","year":"2022","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref039","doi-asserted-by":"crossref","first-page":"1612","DOI":"10.2353\/ajpath.2006.060555","article-title":"Heme oxygenase-2 is a critical determinant for execution of an acute inflammatory and reparative response","volume":"169","author":"F Seta","year":"2006","journal-title":"American J. Path"},{"key":"pone.0290562.ref040","doi-asserted-by":"crossref","first-page":"3422","DOI":"10.3168\/jds.2006-811","article-title":"Dietary L-carnitine affects periparturient nutrient metabolism and lactation in multiparous cows","volume":"90","author":"DB Carlson","year":"2007","journal-title":"J. Dairy Sci"},{"key":"pone.0290562.ref041","doi-asserted-by":"crossref","first-page":"1722","DOI":"10.3168\/jds.S0022-0302(03)73758-8","article-title":"Liver metabolism and production of cows fed increasing amounts of rumen-protected choline during the periparturient period","volume":"86","author":"MS Piepenbrink","year":"2003","journal-title":"J. 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