饲粮添加酿酒酵母和地衣芽孢杆菌对绵羊生长性能与瘤胃发酵的影响

doi:10.3864/j.issn.0578-1752.2020.16.015

摘要/Abstract

摘要：

【目的】研究饲粮中添加酿酒酵母和地衣芽孢杆菌对绵羊生长性能和瘤胃发酵的影响,为酿酒酵母和地衣芽孢杆菌在肉羊饲粮中的合理应用提供理论依据。【方法】试验选用48只4月龄、体重相近（22.96±2.00）kg、健康的杜泊×小尾寒羊杂交F₁代公羔,根据饲喂添加剂不同随机分为4组：D（对照组,两种菌均不添加）;D1（酿酒酵母,6×10¹⁰CFU/kg）;D2（地衣芽孢杆菌,2×10¹⁰CFU/kg）;D3（酿酒酵母6×10¹⁰CFU/kg+地衣芽孢杆菌2×10¹⁰CFU/kg）,每组12只羊,试验期共75 d,前15 d为适应期,正饲期60d。试验羊每天分别于08:00和18:00进行饲喂,自由采食和饮水。正饲期内每天准确称量记录每只试验羊的喂料量和剩料量,并在第1、30、60 天晨饲前对试验羊进行称重,计算平均日采食量、平均日增重（ADG）及料重比（F/G）。试验结束当天08：00正常饲喂试验羊,3 h后采集瘤胃液,测定发酵参数、消化酶活性以及功能微生物。【结果】1）饲粮中添加酿酒酵母和地衣芽孢杆菌对试验羊的初始体重和终末体重及平均采食量的影响均不显著（P>0.05）,D3组的ADG显著高于D组（P<0.05）,D3组的F/G显著低于D组（P<0.05）,D1和D2组的F/G差异不显著（P>0.05）;2）饲粮中添加酿酒酵母和地衣芽孢杆菌对瘤胃液pH、丁酸浓度及乙丙比的影响均不显著（P>0.05）,D3组的NH₃-N浓度显著低于D组（P<0.05）,TVFA和丙酸浓度显著高于D组（P<0.05）,且D3组与D1和D2组均差异不显著（P>0.05）,D3和D2组的乙酸浓度显著高于D1与D组（P<0.05）;3）D3组的β-葡萄糖苷酶、羟甲基纤维素酶、木聚糖酶及淀粉酶活性均显著高于其他3组（P<0.05）。D1和D2组的β-葡萄糖苷酶、果胶酶、羟甲基纤维素酶及淀粉酶活性与D组无显著差异（P>0.05）。D3组蛋白酶活性显著高于D和D2组（P<0.05）,与D1组差异不显著（P>0.05）,D1和D2组的β-葡萄糖苷酶、果胶酶、羟甲基纤维素酶、木聚糖酶、蛋白酶及淀粉酶活性均差异不显著（P>0.05）;4）添加酿酒酵母和地衣芽孢杆菌对黄色瘤胃球菌、栖瘤胃普雷沃氏菌、嗜淀粉瘤胃杆菌和原虫数量的影响不显著（P>0.05）,D3组的溶纤维丁酸弧菌数显著高于其他3组（P<0.05）,白色瘤胃球菌和产琥珀酸丝状杆菌显著高于D组（P<0.05）,但与D1和D2组差异不显著（P>0.05）。与对照组相比,试验组产甲烷菌数量显著降低（P<0.05）,其中D3组最低。【结论】饲粮中添加6×10¹⁰CFU/kg酿酒酵母和2×10¹⁰CFU/kg地衣芽孢杆菌均会对绵羊瘤胃发酵产生积极影响,提高了瘤胃消化酶的活性,增加了瘤胃有益菌的数量,且二者组合饲喂效果更加显著。

关键词: 酿酒酵母, 地衣芽孢杆菌, 瘤胃发酵, 酶活性, 瘤胃微生物区系, 绵羊

Abstract:

【Objective】The objective of this experiment was to study the effects of Saccharomyces cerevisiae and Bacillus licheniformis on growth performance and rumen fermentation in sheep. 【Method】A total of 48 four-month-old healthy Dorper×Small Tail Han sheep ram lambs with similar body weight (22.96±2.00 kg) were randomly selected into 4 groups: D (the control group, without both bacteria); D1 (only adding Saccharomyces cerevisiae, 6×10¹⁰CFU/kg), D2 (only adding Bacillus licheniformis, 2×10¹⁰CFU/kg), and D3 (Saccharomyces cerevisiae 6×10¹⁰CFU/kg + Bacillus licheniformis 2×10¹⁰CFU/kg), with 12 sheep of each group. The experiment was conducted over 75d, with a 15d adaptation and a 60d the formal period. The experimental sheep were fed 2 times per day (08:00 a.m., 18:00 p.m.), free ingestion and free drinking water. During the normal feeding period, each experimental sheep were recorded accurately the amount of feed and residual of weight every day. The experimental sheep were weighed before the morning feeding on the 1st, 30th and 60th day, and the average daily intake, average daily gain (ADG) and Feed/Gain (F/G) value were calculated. At 08:00 on the day of the end of the experiment, the experimental sheep were fed normally. After 3 hours, the rumen fluid was collected and the rumen fermentation, digestive enzyme activities, and functional microorganisms were determined. 【Result】The results showed as follows: (1) the addition of Saccharomyces cerevisiae and Bacillus licheniformis in the diet had no significant effect on the final body weight and average feed intake of the test sheep (P>0.05). The ADG of D3 group was significantly higher than that of D group (P<0.05), and F/G value of D3 group was significantly lower than that of D group (P<0.05). There was no significant difference between D1 and D2 groups in F/G value (P>0.05): (2) the addition of Saccharomyces cerevisiae and Bacillus licheniformis in the diet had no significant effect on the rumen pH value, the concentration of butyric acid, and the value of Acetate/Propionate (A/P). The concentration of NH₃-N for D3 group was significantly lower than D group, but the concentration of TVFA and propionic acid was significantly higher than D group (P<0.05), with no difference among three experimental groups (P>0.05). The concentration of acetic acid in D3 and D2 group was significantly higher than that in D1 and D groups (P<0.05).3)The activity of β-glucosidase, carboxymethyl cellulose, xylanase, and amylase for D3 group was significantly higher than the other groups (P<0.05). There was no significant effect in the activities of β - glucosidase, pectase, carboxymethyl cellulose and amylase among D , D1 and D2 groups (P>0.05). The activity of protease in D3 group was significantly higher than D and D2 groups (P<0.05), with no significant difference between D3 and D1 groups(P>0.05). There was no significant difference in β-glucosidase, pectase, carboxymathy cellulose, xylanase, protease and amylase activities between D1 and D2 groups (P>0.05); (4) the addition of Saccharomyces cerevisiae and Bacillus licheniformis in the diet had no significant effect on the amount of R. flavefaciens, P. ruminicola, R. amylophilus, and Protozoan(P>0.05). The amount of B.fibrisolvens for D3 group was significantly higher than the other 3 groups (P<0.05). The amount of R. albus and F. succinogensin D3 group were significantly higher than D group (P<0.05), with no significant difference among D1, D2 and D3 groups (P>0.05). Compared with the D group, the amount of Methanogensin experimental groups were significantly decreased (P<0.05), and the D3 group was lowest. 【Conclusion】In conclusion, the dietary supplementation of Saccharomyces cerevisiae (6×10¹⁰CFU/kg) and Bacillus licheniformis (2×10¹⁰CFU/kg) had positive effects on rumen fermentation, which improved rumen digestive enzyme activity and increased the number of beneficial bacteria in rumen. Moreover, the combination of Saccharomyces cerevisiae and Bacillus licheniformis achieved better effect.

Key words: Saccharomyces cerevisiae, Bacillus licheniformis, rumen fermentation, enzyme activity, rumen microflora, sheep

郑玮才,郝小燕,张宏祥,项斌伟,张文佳,张春香,张建新. 饲粮添加酿酒酵母和地衣芽孢杆菌对绵羊生长性能与瘤胃发酵的影响[J]. 中国农业科学, 2020, 53(16): 3385-3393.

ZHENG WeiCai,HAO XiaoYan,ZHANG HongXiang,XIANG BinWei,ZHANG WenJia,ZHANG ChunXiang,ZHANG JianXin. Effects of Saccharomyces Cerevisiae and Bacillus Licheniformis on Growth Performance and Rumen Fermentation in Sheep[J]. Scientia Agricultura Sinica, 2020, 53(16): 3385-3393.

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项目 Item		营养水平 Nutrient level
原料 Ingredients		干物质 DM	94.99
玉米 Corn	30.00	粗蛋白质 CP	13.14
米糠 Rice bran	7.00	粗灰分 Ash	8.27
豆粕 Soybean meal	18.00	粗脂肪 EE	2.23
预混料 Premix¹	5.00	中性洗涤纤维 NDF	41.84
玉米秸秆 Corn straw	20.00	酸性洗涤纤维 ADF	22.29
谷草 Millet straw	20.00	钙 Ca	0.35
合计 Total	100.00	磷 P	0.29

基因 Gene name	引物序列（5'→3'）Primer sequence(5'→3')	产物大小 Product/bp	参考文献 References
总菌 Total bacterial	F:CGGTGAATACGTTCYCGG	123	DENMAN等^[14]
总菌 Total bacterial	R:GGWTACCTTGTTACGACTT	123	DENMAN等^[14]
黄色瘤胃球菌 flavefaciens	F:ATTGTCCCAGTTCAGATTGC	173	DENMAN等^[14]
黄色瘤胃球菌 flavefaciens	R:GGCGTCCTCATTGCTGTTAG	173	DENMAN等^[14]
白色瘤胃球菌 albus	F:CCCTAAAAGCAGTCTTAGTTCG	176	KOIKE等^[15]
白色瘤胃球菌 albus	R:CCTCCTTGCGGTTAGAACA	176	KOIKE等^[15]
产琥珀酸丝状杆菌 succinogens	F:GGCGGGATTGAATGTACCTTGAGA	204	DENMAN等^[14]
产琥珀酸丝状杆菌 succinogens	R:TCCGCCTGCCCCTGAACTATC	204	DENMAN等^[14]
溶纤维丁酸弧菌 fibrisolvens	F:TAACATGAGAGTTTGATCCTGGCTC	136	FORSTER等^[16]
溶纤维丁酸弧菌 fibrisolvens	R:CGTTACTCACCCGTCCGC	136	FORSTER等^[16]
嗜淀粉瘤胃杆菌 R. amylophilus	F:CTGGGGAGCTGCCTGAATG	100	JAMI等^[17]
嗜淀粉瘤胃杆菌 R. amylophilus	R:GCATCTGAATGCGACTGGTTG	100	JAMI等^[17]
栖瘤胃普雷沃氏菌 P. ruminicola	F:GAAAGTCGGATTAATGCTCTATGTTG	74	JAMI等^[17]
栖瘤胃普雷沃氏菌 P. ruminicola	R:CATCCTATAGCGGTAAACCTTTGG	74	JAMI等^[17]
产甲烷菌 Methanogens	F:TTCGGTGGATCDCARAGRGC	140	ZHANGI等^[18]
产甲烷菌 Methanogens	R:GBARGTCGWAWCCGTAGAATCC	140	ZHANGI等^[18]
原虫 Protozoan	F:GCTTTCGWTGGTAGTGTATT	223	ZHANGI等^[18]
原虫 Protozoan	R:CTTGCCCTCYAATCGTWCT	223	ZHANGI等^[18]

项目 Items	组别Groups				SEM	P值 P value
项目 Items	D	D1	D2	D3	SEM	P值 P value
初始体重IBW (kg)	23.08	22.22	23.58	22.97	1.20	0.73
终末体重FBW (kg)	39.91	40.04	40.92	43.23	2.77	0.61
平均日增重ADG (g·d^-1)	280.42d	297.38b	289.34c	337.40a	35.22	0.01
平均日采食量ADFI (g·d^-1)	1827.11	1768.90	1759.31	1705.04	165.49	0.91
料重比F/G	6.57a	5.98ab	6.02ab	5.01b	0.27	0.02

项目 Items	组别Groups				SEM	P值 P value
项目 Items	D	D1	D2	D3	SEM	P值 P value
pH	6.76	6.58	6.52	6.74	0.14	0.11
氨态氮NH₃-N (mg·dL^-1)	25.09a	22.94ab	23.02ab	19.87b	1.60	0.03
总挥发性脂肪酸TVFA (mmol·L^-1)	48.03b	51.81ab	54.37a	54.86a	2.22	0.02
乙酸Acetate (mmol·L^-1)	35.25c	38.01b	39.69a	40.09a	0.58	<0.001
丙酸Propionate (mmol·L^-1)	11.44b	12.07ab	12.99a	13.39a	0.51	0.004
丁酸Butyrate (mmol·L^-1)	6.67	6.94	6.78	6.92	0.66	0.97
乙丙比Acetate/Propionate	3.08	3.15	3.07	3.01	0.13	0.69

项目 Items	组别Groups				SEM	P值 P value
项目 Items	D	D1	D2	D3	SEM	P值 P value
β-葡萄糖苷酶β-glucosidase	0.08b	0.09b	0.09b	0.17a	0.01	<0.001
果胶酶Pectase	0.62b	0.73ab	0.88b	0.93a	0.08	0.002
羟甲基纤维素酶Carboxymethyl cellulose	0.09b	0.15b	0.16b	0.25a	0.03	0.001
木聚糖酶 Xylanase	0.23c	0.26bc	0.29b	0.46a	0.01	<0.001
蛋白酶 Protease	15.77c	19.54ab	17.43bc	21.21a	0.11	<0.001
淀粉酶Amylase	0.24b	0.25b	0.26b	0.30a	0.01	0.002