亚急性瘤胃酸中毒耐受性不同的奶牛血浆和乳中脂肪酸及代谢物组成分析

doi:10.11686/cyxb2020482

摘要/Abstract

摘要：

为比较研究对亚急性瘤胃酸中毒（SARA）耐受性不同的奶牛血浆和乳中代谢组的差异，试验选用12头装有永久性瘤胃瘘管的泌乳中期荷斯坦奶牛［泌乳天数=（114±22） d］，饲喂精粗比为4∶6的全混合日粮。在试验期第20和21天晨饲后0、2、4、6、8和12 h监测瘤胃pH值，在早晚饲喂前收集奶样，晨饲6 h后采集颈静脉血，用于测定脂肪酸及代谢物组成。基于奶牛瘤胃平均pH值高低，将奶牛分为SARA易感组（SUS，瘤胃pH=5.76，n=4）和SARA耐受组（TOL，瘤胃pH=6.10，n=4）。脂肪酸测定结果分析显示，与TOL组相比，SUS组奶牛血液及乳中的碳链长度≤C16的脂肪酸含量显著增高（P<0.05），而>C16的脂肪酸的含量显著降低（P<0.05），乳中饱和脂肪酸及单不饱和脂肪酸含量均显著降低（P<0.05）。血浆代谢组结果显示， SUS组奶牛血浆中L-苯丙酮酸水平显著降低（P<0.05），而MG（18：0/0：0/0：0）、9-HODE、12（13） Ep-9-KODE、烟酰胺、异戊基肉碱、磷酸肌酸和L-谷氨酸水平显著升高（P<0.05）。乳代谢组分析结果显示，与TOL组比较，SUS组奶牛乳中的1-硬脂酰磷酸甘油丝氨酸和鞘氨醇水平显著升高（P<0.05），而甘油磷酰基乙醇胺、3-磷酸甘油和乳清酸等其他14个差异代谢物水平显著降低（P<0.05），这些差异代谢物主要涉及甘油磷脂代谢、甘油脂代谢及泛酸与辅酶A代谢等代谢途径。综上所述，SARA耐受性不同的奶牛的血浆和乳中脂肪酸及代谢物组成存在较大差异，SUS组奶牛血浆中氨基酸代谢活动增加，乳腺的乳脂从头合成能力较强，而从血液中摄取长链脂肪酸能力减弱，与此同时，易感奶牛乳中磷脂、乳清酸等营养物质水平下降，乳品质下降。

关键词: 亚急性瘤胃酸中毒, 耐受性, 奶牛, 脂肪酸, 代谢组

Abstract:

The present study aimed to explore the differences in plasma and milk metabolism of dairy cows with different subacute rumen acidosis （SARA） tolerance. Twelve fistulated Holstein dairy cows in mid lactation ［days in milk=（114±22） d］ were selected and fed the same diet consisting of 60% forage and 40% concentrate mixture. On days 20 and 21 of the trial period， the rumen pH was monitored at 0， 2， 4， 6， 8， and 12 h after morning feeding. Milk samples were collected before feeding in the morning and evening. Blood samples were collected from the jugular vein 6 h after morning feeding. All the milk and blood samples were stored in liquid nitrogen for subsequent fatty acid composition and metabolome analysis. Cows with the lowest and highest ruminal pH were assigned into the susceptible （SUS， n=4） and tolerant （TOL， n=4） groups， respectively. The fatty acid composition results showed that the proportions of ≤C16 fatty acids in the blood and milk were significantly increased in the SUS group （P<0.05）， while the proportions of >C16 fatty acids were significantly decreased （P<0.05）. The levels of the saturated fatty acids （SFA） and monounsaturated fatty acids （MUFA） in the milk were significantly decreased in the SUS group （P<0.05）. The LC-MS metabolomics results revealed that the L-phenylpyruvate was significantly reduced， while the levels of remaining MG （18：0/0：0/0：0）， 9-HODE， niacinamide， isovalerylcarnitine， phosphocreatine， L-glutamic acid， and 12（13） Ep-9-KODE were substantially increased in the SUS group. The results of milk metabolome analysis indicated that the levels of 1-stearoylphosphoglycerolserine and sphingosine were significantly increased， while the levels of 14 different metabolites were significantly decreased in the SUS group （P<0.05）， including glyceryl-phosphoryl-ethanolamine， glycerol 3-phosphate， and orotic acid. These differential metabolites were mainly involved in glycerophospholipid metabolism （P<0.05）， glycerolipid metabolism， and pantothenate and CoA biosynthesis. In conclusion， the fatty acids and metabolic profiles in the plasma and milk of cows with different tolerance to SARA were different. For the SUS group， the amino acid metabolism and the de novo synthesis ability of milk fat was enhanced in the plasma， while the ability to absorb long chain fatty acids from blood was weakened. At the same time， the levels of phospholipid， orotic acid and other nutrients were decreased in the milk， which indicated that the quality of milk was reduced for the SUS cows.

Key words: subacute ruminal acidosis, tolerance, cows, fatty acid, metabolome

张涛, 牟英玉, 亓王盼, 郭长征, 张继友, 毛胜勇. 亚急性瘤胃酸中毒耐受性不同的奶牛血浆和乳中脂肪酸及代谢物组成分析[J]. 草业学报, 2021, 30(7): 101-110.

Tao ZHANG, Ying-yu MU, Wang-pan QI, Chang-zheng GUO, Ji-you ZHANG, Sheng-yong MAO. Analysis of plasma and milk fatty acid and metabolite composition in lactating dairy cows with differing tolerance to subacute ruminal acidosis[J]. Acta Prataculturae Sinica, 2021, 30(7): 101-110.

图/表 7

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营养成分Nutrient compsition	含量Content (%)	营养水平Nutrient levels	含量Content
苜蓿草Alfalfa	24.00	粗蛋白质Crude protein (CP, %)	16.16
燕麦Oat	24.00	中性洗涤纤维Neutral detergent fiber (NDF, %)	36.14
玉米青贮Corn silage	12.00	非纤维性碳水化合物Non-fibrous carbohydrate^2） (NFC, %)	38.68
玉米Corn	19.40	粗灰分Ash (%)	5.97
豆粕Soybean meal	13.50	钙Ca (%)	1.14
玉米酒精糟DDGS	3.80	磷P (%)	0.52
石粉Limestone	0.80	粗脂肪Ether extract (EE, %)	3.05
磷酸氢钙CaHPO₄	1.10	淀粉Starch (%)	17.96
食盐NaCl	0.40	泌乳净能Net energy^3） (MJ·kg^-1)	1.57
预混料Premix^1）	1.00	NFC/NDF	1.07
总量Total	100.00

营养成分Nutrient compsition	含量Content (%)	营养水平Nutrient levels	含量Content
苜蓿草Alfalfa	24.00	粗蛋白质Crude protein (CP, %)	16.16
燕麦Oat	24.00	中性洗涤纤维Neutral detergent fiber (NDF, %)	36.14
玉米青贮Corn silage	12.00	非纤维性碳水化合物Non-fibrous carbohydrate^2） (NFC, %)	38.68
玉米Corn	19.40	粗灰分Ash (%)	5.97
豆粕Soybean meal	13.50	钙Ca (%)	1.14
玉米酒精糟DDGS	3.80	磷P (%)	0.52
石粉Limestone	0.80	粗脂肪Ether extract (EE, %)	3.05
磷酸氢钙CaHPO₄	1.10	淀粉Starch (%)	17.96
食盐NaCl	0.40	泌乳净能Net energy^3） (MJ·kg^-1)	1.57
预混料Premix^1）	1.00	NFC/NDF	1.07
总量Total	100.00

时间 Time (min)	流速 Flow rate (mL·min^-1)	A (%)	B (%)
0.0	0.3	95	5
1.0	0.3	95	5
2.0	0.3	60	40
7.0	0.3	20	80
11.0	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)	A (%)	B (%)
0.0	0.3	95	5
1.0	0.3	95	5
2.0	0.3	60	40
7.0	0.3	20	80
11.0	0.3	5	95
15.5	0.3	95	5
19.5	0.3	95	5

脂肪酸 Fatty acid (FA)	组别 Groups		标准误差 SEM	P值 P-value	脂肪酸 Fatty acid (FA)	组别 Groups		标准误差 SEM	P值 P-value
脂肪酸 Fatty acid (FA)	SUS	TOL	标准误差 SEM	P值 P-value	脂肪酸 Fatty acid (FA)	SUS	TOL	标准误差 SEM	P值 P-value
C4:0	3.67	3.62	0.32	0.891	C20:0	20.84	24.96	0.90	0.008
C6:0	3.19	4.23	0.47	0.032	C18:3n6	0.29	0.38	0.09	0.252
C8:0	1.17	1.43	0.13	0.149	C20:1	0.33	1.13	0.06	<0.001
C10:0	0.39	0.29	0.09	0.241	C18:3n3	0.35	0.09	0.02	<0.001
C11:0	0.43	0.31	0.13	0.299	C21:0	0.99	1.56	0.06	<0.001
C13:0	0.70	0.97	0.12	0.112	C20:2	0.14	0.18	0.05	0.282
C14:0	2.57	1.94	0.36	0.718	C22:0	0.24	0.21	0.08	0.522
C15:0	0.22	0.36	0.06	0.220	C22:1n9	0.40	0.28	0.05	0.114
C15:1	0.52	0.60	0.11	0.104	C20:3n3	0.33	0.22	0.04	0.138
C16:0	32.04	29.27	0.65	0.002	C20:4n6	0.60	0.74	0.06	0.015
C16:1	0.35	0.41	0.10	0.646	C24:1	2.53	1.45	0.26	0.004
C17:0	0.31	0.31	0.08	0.498	C22:6n	0.03	1.10	0.31	0.057
C17:1	0.88	0.04	0.05	<0.001	SFA	88.00	88.77	0.75	0.371
C18:0	21.31	19.32	0.58	0.046	MUFA	7.84	6.52	0.78	0.188
C18:1n9t	0.45	0.35	0.10	0.915	PUFA	3.67	3.94	0.26	0.330
C18:1n9c	2.79	2.67	0.72	0.820	≤C16:0	45.04	43.42	0.55	0.025
C18:2n6t	1.79	1.06	0.16	0.008	>C16:0	54.83	56.22	0.49	0.048
C18:2n6c	0.12	0.16	0.04	0.458