胍基乙酸和甜菜碱对羔羊生长性能、瘤胃发酵和血液代谢的影响

doi:10.3864/j.issn.0578-1752.2023.04.014

摘要/Abstract

摘要：

【目的】研究饲粮中添加胍基乙酸和甜菜碱对羔羊生长性能、瘤胃发酵及血清生化指标的影响，为胍基乙酸和甜菜碱在反刍动物生产中的应用提供理论依据。【方法】试验选用48只体重相近（22.03±1.30）kg、3月龄左右的杜泊×小尾寒羊杂种公羔，采用2×2因子完全随机试验设计，试验因素为胍基乙酸（guanidineacetic acid，GAA）（0或900 mg·kg^-1）和甜菜碱（betaine，BT）（0或5 g·d^-1）。试验羔羊随机分为4组，每组12只。分别为对照组、GAA组、BT组和GAA+BT组。试验期共75 d，其中预试期15 d，正试期60 d。正试期第1天晨饲前称量体重作为初始体重（IBW），正试期内记录每只羊的日采食量、剩料量；正试期结束当日晨饲前先采血后空腹称重，作为终末体重（finwal body eight，FBW），计算平均日增重（ADG）、平均日采食量（ADFI）和饲料效率（FE）；血清用于肌酸代谢指标、同型半胱氨酸（Hcy）和乳酸含量测定。试验结束当天正常饲喂试验羊，3h后采集瘤胃液，测定瘤胃发酵参数、消化酶活性以及功能微生物菌群定量。【结果】（1）饲粮中添加GAA日增重（ADG）较对照组有所提高，但无显著差异（P>0.05），添加BT则显著提高了ADG（P<0.05）。饲粮中添加GAA或BT显著提高了饲料效率（FE）（P<0.05），且饲粮中添加GAA和BT对ADG和FE有显著互作效应（P<0.05）。（2）饲粮中添加GAA或BT显著降低了瘤胃液pH（P<0.05），显著提高羔羊采食后3 h的丙酸和总挥发性脂肪酸浓度（P<0.05），GAA和BT对羔羊瘤胃发酵无显著互作效应（P>0.05）。（3）饲粮中添加GAA或BT显著提高羔羊瘤胃液中黄色瘤胃球菌、产琥珀酸丝状杆菌、溶纤维丁酸弧菌和栖瘤胃普雷沃氏菌的数量（P<0.05），显著降低了瘤胃原虫的数量（P<0.05）。饲粮中添加BT显著降低了产甲烷菌的数量（P<0.05），并有提高嗜淀粉瘤胃杆菌数量的趋势（P=0.098），添加GAA则有降低瘤胃产甲烷菌数量的趋势（P=0.085）。GAA和BT对羔羊瘤胃主要功能微生物菌群无显著互作效应（P>0.05）。（4）饲粮中添加GAA或BT显著提高瘤胃蛋白酶活性（P<0.05），添加BT则显著提高羧甲基纤维素酶、木聚糖酶和α-淀粉酶活性（P<0.05），且有提高纤维二糖酶活性的趋势（P=0.075）。饲粮中添加GAA显著提高瘤胃果胶酶活性（P<0.05），且有提高α-淀粉酶活性的趋势（P=0.056）。添加GAA和BT对果胶酶活性有显著互作效应（P<0.05）。（5）饲粮中添加GAA或BT显著提高血清中肌酸含量（P<0.05）。添加GAA有提高血清中同型半胱氨酸的趋势（P=0.053），有降低L-精氨酸-甘氨酸脒基转移酶（AGAT）的趋势（P=0.056）。添加BT可显著降低血清中同型半胱氨酸浓度（P<0.05），并有提高肌酸激酶浓度的趋势（P=0.063）。饲粮中添加GAA和BT对羔羊血清生化指标无显著互作效应（P>0.05）。【结论】饲粮中添加GAA或BT可以促进瘤胃发酵，增加瘤胃功能微生物菌群数量，提高瘤胃微生物酶活性，提高血清中肌酸含量，并降低同型半胱氨酸的浓度。但与单独添加GAA或BT相比，GAA和BT组合添加对羔羊的生长性能没有进一步的提高。

关键词: 胍基乙酸, 甜菜碱, 羔羊, 生长性能, 瘤胃发酵

Abstract:

【Objective】This study was conducted to investigate the effects of dietary guanidinoacetic acid (GAA) and betaine (BT) on growth performance, rumen fermentation and blood metabolites in lambs, so as to provide a support for applying GAA and BT on ruminants. 【Method】Forty-eight 3-month-old Dorper×Thin-tailed Han F₁ hybrid male lambs with similar body weight (22.03±1.3 kg; mean±SD) were randomly divided into 4 groups with a 2×2 factorial arrangement, and the experimental factors were GAA (0 or 900 mg/kg) and BT (0 or 5 g/day). Lambs in four groups were control group, GAA group, BT group and GAA+BT group, respectively. The feeding experiment lasted for 75 days with 15 days of adaptation. On the first day of the formal experiment period, the body weight (BW) before morning feeding was weighed as the initial BW, and each lamb were recorded accurately the amount of feed and residual of weight every day; at the end of the trial period, the blood samples were collected from the jugular vein, and final body weight (FBW) was weighed before morning feeding to calculate average daily gain (ADG), average daily feed intake (ADFI) and feed efficiency (FE). The blood serum was used for the determination of creatine metabolism indexes, homocysteine and lactate content. The rumen fluid was collected from all lambs at 3 hours after feeding at the end of the experiment, which were used to measured rumen fermentation parameters, digestive enzyme activity, and functional microorganisms. 【Result】(1) Compared to the control group, the addition of GAA in the diet did not significantly increase ADG (P>0.05), while the addition of BT increased ADG significantly (P<0.05). The addition of GAA or BT in the diet increased FE significantly (P<0.05), and GAA and BT had significant interaction on ADG and FE (P<0.05). (2) The rumen pH was decreased by addition of GAA or BT (P<0.05), and the concentration of propionic acid and total volatile fatty acid were increased by GAA or BT (P<0.05). There was no significant interaction between GAA and BT on rumen fermentation in lambs (P>0.05). (3) The addition of GAA or BT increased the populations of R. flavefaciens, F. succinogenes, B. fibrisolvens, and P. ruminicola (P<0.05), and decreased the populations of total protozoa (P<0.05). The addition of BT decreased the populations of total methanogens and had a tendency to increase the populations of Rb.Amylophilus (P<0.05, P=0.098). GAA had a tendency to decrease the populations of total methanogens (P=0.088). The addition of GAA and BT had no significant interaction effect on ruminal main functional microflora in lambs (P>0.05). (4) The activity of protease was increased by addition of GAA and BT (P<0.05). The addition of BT increased carboxymethyl-cellulase, xylanase and α-amylase activities (P<0.05), and had a tendency to increase cellobiase activity (P=0.075). GAA increased pectase activity (P<0.05), and had a tendency to increase α-amylase activitiy (P=0.056). The significant interaction effect between GAA and BT was observed on pectase (P<0.05). (5) The addition of GAA or BT increased the content of creatine in blood serum (P<0.05). The addition of GAA had a tendency to increase the concentration of homocysteine (Hcy) and to decrease L-arginine-glycine amidinotransferase (AGAT) (P=0.053, P=0.056). BT decreased the content of Hcy in blood serum, and had a tendency to increase creatine kinase (P=0.063). GAA and BT had no interactive significant effects on blood metabolites (P>0.05). 【Conclusion】In conclusion, the addition of GAA or BT promoted rumen fermentation, increased the populations of functional rumen microflora and microbial enzyme activity, elevated serum creatine levels, and decreased the content of homocysteine. However, the combination addition of GAA and BT did not further increase the performance of lambs when compared with the addition of GAA or BT alone.

Key words: guanidinoacetic acid, betaine, lambs, growth performance, rumen fermentation

任国栋, 郝小燕, 张暄梓, 刘森, 张宏祥, 田光元, 张建新. 胍基乙酸和甜菜碱对羔羊生长性能、瘤胃发酵和血液代谢的影响[J]. 中国农业科学, 2023, 56(4): 766-778.

REN GuoDong, HAO XiaoYan, ZHANG XuanZi, LIU Sen, ZHANG HongXiang, TIAN GuangYuan, ZHANG JianXin. Effects of Guanidinoacetic Acid and Betaine Supplementation on Growth Performance, Rumen Fermentation and Blood Metabolites in Lambs[J]. Scientia Agricultura Sinica, 2023, 56(4): 766-778.

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原料 Ingredient	含量Content	营养水平 Nutrient level ²⁾
豆粕 Soybean meal	6.00	干物质 DM	93.41
棉籽粕 Cottonseed meal	4.00	粗蛋白质 CP	15.00
玉米胚芽粕 Corn germ meal	15.00	粗脂肪 EE	2.01
喷浆玉米皮 Sprayed corn husk	7.00	粗灰分 Ash	9.13
玉米 Corn	33.00	中性洗涤纤维 NDF	31.68
花生壳粉 Peanut shell powder	12.00	酸性洗涤纤维 ADF	18.03
葵花皮粉 Sunflower peel powder	12.00	钙 Ca	1.19
玉米秸秆 Corn straw	6.00	磷 P	0.48
预混料 Premix¹⁾	5.00	代谢能 ME/(MJ·kg^-1·kg^-1)	9.98
总计Total	100.00

基因 Gene name	引物序列Primer sequence(5'→3')	产物大小 Product (bp)	参考文献 Reference
总菌 Total bacterial	F:CGGTGAATACGTTCYCGG R:GGWTACCTTGTTACGACTT	123	DENMAN et al.^[28]
总厌氧真菌 Total anaerobic fungi	F:GAGGAAGTAAAAGTCGTAACAAGGTTTC R:CAAATTCACAAAGGGTAGGATGATT	120	LI et al.^[10]
黄色瘤胃球菌 Flavefaciens	F:ATTGTCCCAGTTCAGATTGC R:GGCGTCCTCATTGCTGTTAG	173	DENMAN et al.^[28]
白色瘤胃球菌 Albus	F:CCCTAAAAGCAGTCTTAGTTCG R:CCTCCTTGCGGTTAGAACA	176	KOIKE et al.^[29]
产琥珀酸丝状杆菌 Succinogens	F:GGCGGGATTGAATGTACCTTGAGA R:TCCGCCTGCCCCTGAACTATC	204	DENMAN et al.^[28]
溶纤维丁酸弧菌 Fibrisolvens	F:TAACATGAGAGTTTGATCCTGGCTC R:CGTTACTCACCCGTCCGC	136	FORSTER et al.^[30]
嗜淀粉瘤胃杆菌 R. amylophilus	F:CTGGGGAGCTGCCTGAATG R:GCATCTGAATGCGACTGGTTG	100	JAMI et al.^[31]
栖瘤胃普雷沃氏菌 P. ruminicola	F:GAAAGTCGGATTAATGCTCTATGTTG R:CATCCTATAGCGGTAAACCTTTGG	74	JAMI et al.^[31]
产甲烷菌 Methanogens	F:TTCGGTGGATCDCARAGRGC R:GBARGTCGWAWCCGTAGAATCC	140	ZHANG et al.^[32]
原虫 Protozoan	F:GCTTTCGWTGGTAGTGTATT R:CTTGCCCTCYAATCGTWCT	223	ZHANG et al.^[32]

项目 Item	BT-		BT+		SEM	P值 P value
项目 Item	GAA-	GAA+	GAA-	GAA+	SEM	BT	GAA	BT×GAA
初始体重 IBW (kg)	26.56	25.94	25.74	25.02	0.26	0.829	0.677	0.178
终末体重 FBW (kg)	44.80	46.21	46.11	45.10	0.39	0.890	0.786	0.106
平均日增重 ADG (g·d^-1)	305.93Bb	337.11a	349.37Aa	337.15a	6.65	0.004	0.166	0.004
平均日采食量 ADFI (g·d^-1)	1792.18	1672.30	1754.93	1688.42	24.87	0.831	0.072	0.590
饲料效率 FE	0.17b	0.20a	0.19a	0.20a	0.004	0.023	0.011	0.023

项目 Item	BT-		BT+		SEM	P 值 P value
项目 Item	GAA-	GAA+	GAA-	GAA+	SEM	BT	GAA	BT×GAA
pH	6.16	6.00	5.98	5.66	0.062	0.021	0.028	0.411
总挥发酸 Total VFA (mmol·L^-1)	96.52	109.88	101.76	122.35	2.660	0.012	<0.001	0.267
挥发酸组成（%，mol/100mol）
乙酸 Acetate (A)	48.29	50.03	49.73	53.47	1.000	0.234	0.183	0.619
丙酸 Propionate (P)	43.52	50.21	47.91	56.40	1.550	0.047	<0.001	0.719
丁酸 Butyrate	5.96	8.49	8.60	8.77	0.494	0.128	0.157	0.211
戊酸 Valerate	2.80	3.95	1.91	2.40	0.215	<0.001	0.010	0.249
异丁酸 Isobutyrate	0.32	0.29	0.29	0.22	0.019	0.202	0.185	0.507
异戊酸 Isovalerate	0.37	0.35	0.27	0.26	0.015	<0.001	0.630	0.570
乙丙比 A:P	1.00	0.98	0.97	0.97	0.014	0.539	0.871	0.820
氨态氮 NH₃-N (mg/100 mL)	10.50	10.34	10.23	13.10	0.489	0.151	0.186	0.111

项目 Items	BT-		BT+		SEM	P值 P value
项目 Items	GAA-	GAA+	GAA-	GAA+	SEM	BT	GAA	BT×GAA
羧甲基纤维素酶 Carboxymethyl-cellulase	0.36	0.36	0.40	0.38	0.013	0.023	0.609	0.701
纤维二糖酶Cellobiase	0.53	0.59	0.63	0.60	0.016	0.075	0.611	0.176
木聚糖酶 Xylanase	0.67	0.70	0.80	0.86	0.025	0.002	0.274	0.681
果胶酶 Pectinase	2.99B	3.53A	3.79A	3.54A	0.071	<0.001	0.091	<0.001
α-淀粉酶 α-amylase	1.31	1.34	1.54	1.74	0.047	<0.001	0.056	0.158
蛋白酶 Protease	6.26	7.13	7.83	10.70	0.428	<0.001	0.005	0.112