Plasma biochemical indexes and metabolomics profile changes of dairy cows with subacute ruminal acidosis

doi:10.11686/cyxb2020323

Abstract

Abstract:

This research explored the relationship between subacute ruminal acidosis （SARA） and blood metabolic changes in dairy cows. Eight ruminally cannulated Holstein cows in mid-lactation were randomly assigned to either a low-concentrate （40%） diet （CON） or a high-concentrate （60%） diet （SARA） for 3 weeks. On the last day of each week of the experimental period， ruminal pH was measured at 0， 2， 4， 6， 8 and 12 h after morning feeding， and blood samples for metabolomics analysis and determination of the biochemical indexes were obtained from the jugular vein at 6 h after morning feeding. It was found that， compared with the control group， ruminal pH was significantly reduced in the SARA group （P=0.002）. No difference was observed in dry matter intake （DMI） between the two groups （P=0.524）. The results for plasma biochemical indexes showed that cows fed the SARA diet had a lower plasma concentration of β-hydroxybutyrate （P=0.007）， and a higher concentration of triglycerides （P=0.014）. Plasma metabolomics profile was performed using liquid chromatography/mass spectrometry （LC/MS）. Both principal component analysis （PCA） and partial least squares discriminant analysis （PLS-DA） showed a clear separation between the CON and SARA groups. By multivariate analysis， 26 differential plasma metabolites were identified ［variable importance in the projection （VIP） >1 and false discovery rate （FDR）<0.05］. Compared with the CON group， in the SARA group， 7-ketodeoxycholic acid， deoxycholic acid， cholic acid， 12-ketodeoxycholic acid， 12（13）Ep-9-KODE， 12，13-DHOME and L-asparagine metabolites were significantly increased （FDR<0.05）， while undecanedioic acid， hexadecanedioic acid， 9-HODE， thromboxane B3， PGE2， L-arginine， L-tryptophan， L-threonine， isobutyrylglycine， isovalerylglycine， hippuric acid， 4-hydroxyhippuric acid and 6-phospho-2-dehydro-D-gluconate were significantly reduced （FDR<0.05）. Overall， compared with the CON group， SARA affected the lipid metabolism， amino acid metabolism and glucose metabolism of dairy cows. The identified differential metabolites may serve as potential biomarkers for SARA.

Key words: subacute ruminal acidosis, dairy cows, plasma, LC/MS, metabolomics

Wang-pan QI, Ying-yu MU, Tao ZHANG, Ji-you ZHANG, Sheng-yong MAO. Plasma biochemical indexes and metabolomics profile changes of dairy cows with subacute ruminal acidosis[J]. Acta Prataculturae Sinica, 2021, 30(6): 141-150.

Figures/Tables 9

References 29

1	Zebeli Q， Dijkstra J， Tafaj M， et al. Modeling the adequacy of dietary fiber in dairy cows based on the responses of ruminal pH and milk fat production to composition of the diet. Journal of Dairy Science， 2008， 91（5）： 2046-2066.
2	Penner G B， Steele M A， Aschenbach J R， et al. Ruminant nutrition symposium： Molecular adaptation of ruminal epithelia to highly fermentable diets. Journal of Animal Science， 2011， 89（4）： 1108-1119.
3	Humer E， Petri R M， Aschenbach J R， et al. Invited review： Practical feeding management recommendations to mitigate the risk of subacute ruminal acidosis in dairy cattle. Journal of Dairy Science， 2018， 101（2）： 872-888.
4	Mao S Y， Huo W J， Zhu W Y. Microbiome-metabolome analysis reveals unhealthy alterations in the composition and metabolism of ruminal microbiota with increasing dietary grain in a goat model. Environmental Microbiology， 2016， 18（2）： 525-541.
5	Liu J H. The effect of subacute ruminal acidosis on ruminal epithelial barrier function and its underlying mechanism in goats. Nanjing： Nanjing Agricultural University， 2014.
	刘军花. 亚急性瘤胃酸中毒对山羊瘤胃上皮屏障功能的影响及其机制. 南京：南京农业大学， 2014.
6	Zebeli Q， Dunn S M， Ametaj B N. Perturbations of plasma metabolites correlated with the rise of rumen endotoxin in dairy cows fed diets rich in easily degradable carbohydrates. Journal of Dairy Science， 2011， 94（5）： 2374-2382.
7	Guo Y， Xu X， Zou Y， et al. Changes in feed intake， nutrient digestion， plasma metabolites， and oxidative stress parameters in dairy cows with subacute ruminal acidosis and its regulation with pelleted beet pulp. Journal of Animal Science and Biotechnology， 2013， 4（1）： 31.
8	Ametaj B N， Zebeli Q， Saleem F， et al. Metabolomics reveals unhealthy alterations in rumen metabolism with increased proportion of cereal grain in the diet of dairy cows. Metabolomics， 2010， 6（4）： 583-594.
9	Saleem F， Ametaj B N， Bouatra S， et al. A metabolomics approach to uncover the effects of grain diets on rumen health in dairy cows. Journal of Dairy Science， 2012， 95（11）： 6606-6623.
10	Zhang R， Zhu W， Jiang L， et al. Comparative metabolome analysis of ruminal changes in Holstein dairy cows fed low- or high-concentrate diets. Metabolomics， 2017， 13（6）： 1-15.
11	Kenez A， Danicke S， Rollekampczyk U， et al. A metabolomics approach to characterize phenotypes of metabolic transition from late pregnancy to early lactation in dairy cows. Metabolomics， 2016， 12（11）： 165.
12	Zhang H Y， Wu L， Xu C， et al. Plasma metabolomic profiling of dairy cows affected with ketosis using gas chromatography/mass spectrometry. BMC Veterinary Research， 2013， 9（1）： 186.
13	Sun H Z， Wang D M， Wang B， et al. Metabolomics of four biofluids from dairy cows： Potential biomarkers for milk production and quality. Journal of Proteome Research， 2015， 14（2）： 1287-1298.
14	Ametaj B N， Emmanuel D G， Zebeli Q， et al. Feeding high proportions of barley grain in a total mixed ration perturbs diurnal patterns of plasma metabolites in lactating dairy cows. Journal of Dairy Science， 2009， 92（3）： 1084-1091.
15	Sun P F. Effect of forage combination on ruminal fermentation and milk fatty acids composition. Hangzhou： Zhejiang University， 2007.
	孙攀峰. 粗饲料组合对瘤胃发酵及乳脂肪酸组成的影响及其机理研究. 杭州：浙江大学， 2007.
16	Bauman D E， Mather I H， Wall R， et al. Major advances associated with the biosynthesis of milk. Journal of Dairy Science， 2006， 89（4）： 1235-1243.
17	Sun Y Y， Hu H L， Gao M， et al. Effects of subacute ruminal acidosis on plasma abnormal metabolites and biochemical indexes of dairy goats. Chinese Journal of Animal Nutrition， 2017， 29（3）： 1046-1055.
	孙燕勇，胡红莲，高民，等. 亚急性瘤胃酸中毒对奶山羊血浆异常代谢产物及生化指标的影响. 动物营养学报， 2017， 29（3）： 1046-1055.
18	Li X S， Yang F L， Du Y L， et al. Progress on dairy cow ketosis and preventive measures. Progress in Veterinary Medicine， 2010， 31（1）： 108-111.
	李小杉，杨丰利，杜玉兰，等. 奶牛酮病及其预防措施. 动物医学进展， 2010， 31（1）： 108-111.
19	Sun L W， Zhang H Y， Xia C， et al. Serum metabonomic studies of ketosis in dairy cows. Acta Veterinaria et Zootechnica Sinica， 2013， 44（10）： 1667-1674.
	孙玲伟，张洪友，夏成，等. 奶牛酮病的血浆代谢组学分析. 畜牧兽医学报， 2013， 44（10）： 1667-1674.
20	Guo J F. Inflammatory injury in the liver of ruminants by subacute ruminal acidosis （SARA） and manipulation of SARA. Nanjing： Nanjing Agricultural University， 2017.
	郭峻菲. 亚急性瘤胃酸中毒（SARA）对反刍动物肝脏的炎性损伤与SARA的调控研究. 南京：南京农业大学， 2017.
21	Alvarez C， Nelms C， Daddio V， et al. The pancreatic duct epithelium in vitro： Bile acid injury and the effect of epidermal growth factor. Surgery， 1997， 122（2）： 476-484.
22	Li T， Apte U. Bile acid metabolism and signaling in cholestasis， inflammation， and cancer. Advances in Pharmacology， 2015， 74： 263-302.
23	Angelin B， Einarsson K， Hellstrom K， et al. Bile acid kinetics in relation to endogenous tryglyceride metabolism in various types of hyperlipoproteinemia. Journal of Lipid Research， 1978， 19（8）： 1004-1016.
24	Spencer T E， Burghardt R C， Johnson G A， et al. Conceptus signals for establishment and maintenance of pregnancy. Reproductive Biology and Endocrinology， 2004， 2（1）： 49.
25	Guo C， Xue Y， Seddik H E， et al. Dynamic changes of plasma metabolome in response to severe feed restriction in pregnant ewes. Metabolites， 2019， 9（6）： 112.
26	Mccue M D. Starvation physiology： Reviewing the different strategies animals use to survive a common challenge. Comparative Biochemistry and Physiology A-molecular & Integrative Physiology， 2010， 156（1）： 1-18.
27	Toromanovic J， Kovacbesovic E， Sapcanin A， et al. Urinary hippuric acid after ingestion of edible fruits. Bosnian Journal of Basic Medical Sciences， 2008， 8（1）： 38-43.
28	Liu J， Xu T， Liu Y， et al. A high-grain diet causes massive disruption of ruminal epithelial tight junctions in goats. American Journal of Physiology-regulatory Integrative and Comparative Physiology， 2013， 305（3）： 232-241.
29	Malekkhahi M， Tahmasbi A M， Naserian A A， et al. Effects of supplementation of active dried yeast and malate during sub-acute ruminal acidosis on rumen fermentation， microbial population， selected blood metabolites， and milk production in dairy cows. Animal Feed Science and Technology， 2016， 213： 29-43.

项目 Item	对照组 CON	处理组 SARA
日粮组成 Diet ingredient
苜蓿草 Alfalfa (%DM)	24.00	17.00
燕麦草 Oat grass (%DM)	24.00	17.00
青贮玉米 Silage (%DM)	12.00	6.00
玉米 Corn (%DM)	19.40	24.92
豆粕 Soybean meal (%DM)	13.50	13.48
大麦 Barley (%DM)	0.00	12.00
玉米酒精糟 Corn distiller’s grains (%DM)	3.80	5.91
石粉 Limestone (%DM)	0.80	1.48
磷酸氢钙 Calcium monohydrogen phosphate (%DM)	1.10	0.92
食盐 Salt (%DM)	0.40	0.37
预混料 Premix¹ (%DM)	1.00	0.92
营养水平 Nutritional level
粗蛋白 Crude protein (%DM)	16.16	16.12
钙 Ca (%DM)	1.14	1.18
磷 P (%DM)	0.52	0.51
中性洗涤纤维 Neutral detergent fiber (%DM)	36.14	29.92
非纤维性碳水化合物 Non-fibrous carbohydrate (%DM)	35.39	42.34
粗灰分 Crude ash (%DM)	5.97	4.87
泌乳净能 Net energy (Mcal·kg^-1)	1.57	1.64
粗脂肪 Ether extracts (%DM)	3.05	3.05
淀粉 Starch (%DM)	17.96	27.82

项目 Item	对照组 CON	处理组 SARA
日粮组成 Diet ingredient
苜蓿草 Alfalfa (%DM)	24.00	17.00
燕麦草 Oat grass (%DM)	24.00	17.00
青贮玉米 Silage (%DM)	12.00	6.00
玉米 Corn (%DM)	19.40	24.92
豆粕 Soybean meal (%DM)	13.50	13.48
大麦 Barley (%DM)	0.00	12.00
玉米酒精糟 Corn distiller’s grains (%DM)	3.80	5.91
石粉 Limestone (%DM)	0.80	1.48
磷酸氢钙 Calcium monohydrogen phosphate (%DM)	1.10	0.92
食盐 Salt (%DM)	0.40	0.37
预混料 Premix¹ (%DM)	1.00	0.92
营养水平 Nutritional level
粗蛋白 Crude protein (%DM)	16.16	16.12
钙 Ca (%DM)	1.14	1.18
磷 P (%DM)	0.52	0.51
中性洗涤纤维 Neutral detergent fiber (%DM)	36.14	29.92
非纤维性碳水化合物 Non-fibrous carbohydrate (%DM)	35.39	42.34
粗灰分 Crude ash (%DM)	5.97	4.87
泌乳净能 Net energy (Mcal·kg^-1)	1.57	1.64
粗脂肪 Ether extracts (%DM)	3.05	3.05
淀粉 Starch (%DM)	17.96	27.82

时间 Time (min)	流速 Flow rate (mL·min^-1)	流动相A Mobile phase A (%)	流动相B Mobile phase B (%)
0	0.3	95	5
1	0.3	95	5
2	0.3	60	40
7	0.3	20	80
11	0.3	5	95
15.5	0.3	95	5
19.5	0.3	95	5

时间 Time (min)	流速 Flow rate (mL·min^-1)	流动相A Mobile phase A (%)	流动相B Mobile phase B (%)
0	0.3	95	5
1	0.3	95	5
2	0.3	60	40
7	0.3	20	80
11	0.3	5	95
15.5	0.3	95	5
19.5	0.3	95	5

项目 Item	对照组 CON (mmol·L^-1)	试验组 SARA (mmol·L^-1)	标准误 SEM	P值 P-value
项目 Item	对照组 CON (mmol·L^-1)	试验组 SARA (mmol·L^-1)	标准误 SEM	饲喂天数 Feeding days	日粮 Diet	饲喂天数日粮 Daydiet
β-羟丁酸 β-hydroxybutyric acid (BHBA)	0.449	0.393	0.013	0.200	0.007	0.002
葡萄糖 Glucose (Glu)	6.166	6.894	0.224	0.003	0.057	0.867
胆固醇 Cholesterol (TC)	5.323	5.151	0.171	0.691	0.645	0.783
甘油三酯 Triglyceride (TG)	1.228	1.508	0.061	0.064	0.014	0.499
游离脂肪酸Nonestesterified fatty acid (NEFA)	0.153	0.148	0.004	0.284	0.584	0.865