醋酸棉酚对绵羊瘤胃微生物数量及消化代谢的影响
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摘要
本试验旨在研究添加醋酸棉酚后绵羊瘤胃微生物数量、瘤胃液代谢指标、水解酶活性的变化,探究醋酸棉酚对绵羊瘤胃微生物数量及消化代谢的影响。试验选用8只3岁左右、平均体重为(49.13±4.70) kg的健康哈萨克羊,安装永久性瘤胃瘘管后随机分为2组(每组4只),分别为对照组和试验组,试验分为2期,每期25 d。所有羊只饲喂同一营养水平粉状精料,每只羊每天粉状精料饲喂量为体重的1.50%,小麦秸秆及苜蓿为体重的1.00%,玉米青贮为体重的0.05%(干物质基础),混合饲喂,自由饮水;在此基础上,试验Ⅰ期试验组饲粮每天每只添加600 mg醋酸棉酚,试验Ⅱ期饲粮每天每只添加1 200 mg醋酸棉酚。结果表明:添加醋酸棉酚对绵羊瘤胃液pH及细菌总数量无显著影响(P>0.05),饲喂前(即0 h)试验组绵羊瘤胃液氨态氮浓度显著高于对照组(P<0.05),饲喂后6.0、9.0、12.0 h,试验组绵羊瘤胃液挥发性脂肪酸浓度显著低于对照组(P<0.05),试验Ⅰ期各时间点试验组绵羊瘤胃原虫总数量极显著低于对照组(P<0.01),饲喂后3.0 h,试验组绵羊瘤胃液纤维素酶活性显著高于对照组(P <0. 05),饲喂后9. 0、12.0 h,试验组绵羊瘤胃液蛋白酶活性极显著低于对照组(P <0.01);试验Ⅱ期,饲喂后6.0 h,试验组绵羊瘤胃液蛋白酶活性显著低于对照组(P<0.05),饲喂后12.0 h极显著低于对照组(P<0.01);添加醋酸棉酚后瘤胃液中棉酚的浓度在饲喂后1. 5、3. 0 h最高。综上,醋酸棉酚在初期摄入后显著降低绵羊瘤胃原虫数量;醋酸棉酚通过抑制绵羊瘤胃液蛋白酶活性,降低瘤胃液中戊酸、异戊酸及异丁酸的浓度而对瘤胃内蛋白质消化代谢产生影响,这种作用主要发生在摄入醋酸棉酚6.0 h后;醋酸棉酚对绵羊瘤胃液中的纤维素酶、淀粉酶的活性整体没有显著影响。
This experiment aimed to explore the effects of gossypol acetate on rumen microbial population,digestion and metabolism in sheep by analyzing microbial population,rumen fluid metabolism indexes,and activity of some hydrolases in rumen after adding gossypol acetate. Eight Kazakh sheep aged about 3-years-old,with permanent rumen fistula,and an average body weight of( 49.13±4.70) kg,were randomly divided into two groups( control group and trial group,four animals in each group). The experiment was divided into two phases,with 25 d in each phase. All sheep were fed the same powder concentrate,the feeding account of pow-der concentrate was 1.50% of body weight per sheep per day,wheat strawand alfalfa were 1. 00% of body weight,corn silage was 0.05% of body weight( dry matter basis),and mixed feeding and free drinking water.On the basis of this diet,trial group in period Ⅰ and period Ⅱ added gossypol acetate( 600 mg per sheep per day and 1 200 mg per sheep per day,respectively). The results showed that there was no significant difference in the rumen pH after adding gossypol acetate( P > 0. 05). The rumen ammonia nitrogen concentration in the trial group was significantly higher than that in the control group before feeding gossypol acetate( 0 h)( P <0.05). The concentration of volatile fatty acid in the trial group was significantly lower than that in the control group at 6.0,9.0 and 12.0 h after feeding( P<0.05). During the period Ⅰ,the total population of rumen fluid protozoa in the trial group was extremely significantly lower than that in the control group( P < 0.01). The rumen cellulase activity in the trial group was significantly higher than that in the control group at 3. 0 h after feeding( P<0.05). The protease activity was extremely significantly lower than that in the control group at 9.0 and 12.0 h after feeding( P<0.01). During the period Ⅱ,the protease activity in the trial group was significantly lower than that in the control group at 6.0 h after feeding( P<0.05),and extremely significantly lower than that in the control group at 12.0 h after feeding( P < 0.01). After adding gossypol acetate,the gossypol concentration in rumen was the highest at 1.5 and 3.0 h after feeding. Therefore,the rumen protozoa population of sheep significantly reduces after initial intake of gossypol acetate. Gossypol acetate affects protein digestion and metabolism in rumen by inhibiting the protease activity,and reducing the concentration of valeric acid,isovaleric acid and isobutyric acid in rumen fluid. This effect mainly occurs at 6.0 h after intaking gossypol acetate. Gossypol acetate does not have significant effects on rumen fluid cellulase and amylase activities in sheep as a whole.[Chinese Journal of Animal Nutrition,2019,31(7):3421-3430]
This experiment aimed to explore the effects of gossypol acetate on rumen microbial population,digestion and metabolism in sheep by analyzing microbial population,rumen fluid metabolism indexes,and activity of some hydrolases in rumen after adding gossypol acetate. Eight Kazakh sheep aged about 3-years-old,with permanent rumen fistula,and an average body weight of( 49.13±4.70) kg,were randomly divided into two groups( control group and trial group,four animals in each group). The experiment was divided into two phases,with 25 d in each phase. All sheep were fed the same powder concentrate,the feeding account of pow-der concentrate was 1.50% of body weight per sheep per day,wheat strawand alfalfa were 1. 00% of body weight,corn silage was 0.05% of body weight( dry matter basis),and mixed feeding and free drinking water.On the basis of this diet,trial group in period Ⅰ and period Ⅱ added gossypol acetate( 600 mg per sheep per day and 1 200 mg per sheep per day,respectively). The results showed that there was no significant difference in the rumen pH after adding gossypol acetate( P > 0. 05). The rumen ammonia nitrogen concentration in the trial group was significantly higher than that in the control group before feeding gossypol acetate( 0 h)( P <0.05). The concentration of volatile fatty acid in the trial group was significantly lower than that in the control group at 6.0,9.0 and 12.0 h after feeding( P<0.05). During the period Ⅰ,the total population of rumen fluid protozoa in the trial group was extremely significantly lower than that in the control group( P < 0.01). The rumen cellulase activity in the trial group was significantly higher than that in the control group at 3. 0 h after feeding( P<0.05). The protease activity was extremely significantly lower than that in the control group at 9.0 and 12.0 h after feeding( P<0.01). During the period Ⅱ,the protease activity in the trial group was significantly lower than that in the control group at 6.0 h after feeding( P<0.05),and extremely significantly lower than that in the control group at 12.0 h after feeding( P < 0.01). After adding gossypol acetate,the gossypol concentration in rumen was the highest at 1.5 and 3.0 h after feeding. Therefore,the rumen protozoa population of sheep significantly reduces after initial intake of gossypol acetate. Gossypol acetate affects protein digestion and metabolism in rumen by inhibiting the protease activity,and reducing the concentration of valeric acid,isovaleric acid and isobutyric acid in rumen fluid. This effect mainly occurs at 6.0 h after intaking gossypol acetate. Gossypol acetate does not have significant effects on rumen fluid cellulase and amylase activities in sheep as a whole.[Chinese Journal of Animal Nutrition,2019,31(7):3421-3430]
引文
[1]元秀晔,谢全喜,陈振,等.高效降解游离棉酚并改善棉籽粕营养品质的菌株筛选[J].动物营养学报,2017,29(9):3258-3266.
[2]金红春,兰时乐,胡毅,等.棉粕发酵前后营养成分变化研究[J].饲料工业,2011,32(13):19-23.
[3] DODOU K,ANDERSON R J,SMALL D A,et al.Investigations on gossypol:past and present developments[J]. Expert Opinion on Investigational Drugs,2005,14(11):1419-1434.
[4]方琴音.饲粮不同棉籽饼用量对商品蛋鸡生产性能、蛋品质、血液生化指标的影响研究[D].硕士学位论文.雅安:四川农业大学,2004.
[5]吕云峰,王修启,赵青余,等.棉酚在饲料中安全限量及畜产品中残留研究进展[J].中国农学通报,2010,26(24):1-5.
[6] EISELE G R. A perspective on gossypol ingestion in swine[J]. Veterinary&Human Toxicology,1986,28(2):118-120.
[7] ABOU-DONIA MB.Physiological effects and metabolism of gossypol[J]. Residue Reviews,1976,61:125-160.
[8]石彩霞,侯彩云,王海荣,等.游离棉酚对绵羊瘤胃发酵和微生物区系的影响[J].中国饲料,2013(5):16-19.
[9]候彩云.棉酚对绵羊瘤胃微生物种群及纤维物质降解的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学,2012.
[10] KNUTSEN H K,BARREGRD L,BIGNAMI M,et al.Presence of free gossypol in whole cottonseed[J].Efsa Journal,2017,15(7):e04850.
[11]屯妮萨·麦提赛伊迪.不同精粗比日粮、粗料型日粮添喂烟酰胺对绵羊瘤胃微生物代谢的影响[D].硕士学位论文.乌鲁木齐:新疆农业大学,2012.
[12]邓凯平,王锋,马铁伟,等.日粮中添加不同水平紫苏籽对湖羊生长性能、瘤胃发酵及养分表观消化率的影响[J].草业学报,2017,26(5):205-212.
[13]杨舒黎,王加启,胡志勇,等.日粮添加豆油和胡麻油对肉牛瘤胃发酵及主要微生物数量的影响[J].中国农业科学,2007,40(10):2316-2322.
[14]陈程.棉铃虫P450基因CPR-CYP6AE14/CYP9A12的异源共表达及其对棉酚降解作用研究[D].博士学位论文.石河子:石河子大学,2018.
[15]张龙翔,张庭芳,李玲媛.生化实验方法和技术[M].2版.北京:高等教育出版社,1997.
[16] HRISTOV A N,ROPP J K,HUNT C W. Effect of barley and its amylopectin content on ruminal fermentation and bacterial utilization of ammonia-N in vitro[J]. Animal Feed Science and Technology,2002,99(1/2/3/4):25-36.
[17] The PLOS ONE Staff. Correction:predicting in vitro rumen VFA production using CNCPS carbohydrate fractions with multiple linear models and artificial neural networks[J].PLoS One,2015,10(3):e0119740.
[18] SCHNEIDER I C,AMES ML,RASMUSSEN MA,et al.Fermentation of cottonseed and other feedstuffs in cattle rumen fluid[J].Journal of Agricultural and Food Chemistry,2002,50(8):2267-2273.
[19] FRENCH E A,BERTICS S J,ARMENTANO L E.Rumen and milk odd-and branched-chain fatty acid proportions are minimally influenced by ruminal volatile fatty acid infusions[J]. Journal of Dairy Science,2012,95(4):2015-2026.
[20]孙万成,罗毅皓,刘祥军.不同泌乳期牦牛乳中奇数与支链脂肪酸的分布[J].食品科学,2015,36(6):199-201.
[21] VLAEMINCK B,FIEVEZ V,VAN LAAR H,et al.Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates[J].Journal of Animal Physiology and Animal Nutrition,2010,88(11/12):401-411.
[22] NSEREKO V L,MORGAVI D P,RODE L M,et al.Effects of fungal enzyme preparations on hydrolysis and subsequent degradation of alfalfa hay fiber by mixed rumen microorganisms in vitro[J]. Animal Feed Science and Technology,2000,88(3/4):156-170.
[23] FESTENSTEIN G N. Cellulolytic enzymes from sheep-rumen liquor micro-organisms[J]. Biochemical Journal,1958,69(4):562-567.
[24] SUHARTATI F M,SURYAPRATAMA W.Increasing the in vitro enzymatic activity of cellulase and amylase from beef cattle rumen fluid supplemented with Moringa oleifera leaves and sulfur[J]. Pakistan Journal of Nutrition,2017,16(8):571-576.
[25]刘文杰,雒秋江,钟涛,等.添加赖氨酸对小尾寒羊瘤胃消化代谢的影响[J].中国畜牧兽医,2015,42(10):2658-2668.
[26] TRIPATHI MK,RAGHUVANSI K S K,MONDAL D,et al. Effect of Bt-cottonseed meal feeding on performance,fermentation,ciliates population and microbial hydrolytic enzymes in lamb[J].African Journal of Biotechnology,2014,13(3):510-522.
[27] LI F,ZHANG P Y,ZHANG G M,et al.Enhancement of corn stover hydrolysis with rumen fluid pretreatment at different solid contents:effect,structural changes and enzymes participation[J].International Biodeterioration&Biodegradation,2016,119:405-412.
[28] MORGAVI D P,BEAUCHEMIN K A,NSEREKO V L,et al.Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases[J]. Journal of Animal Science,2001,79(6):1621-1630.
[29] REISER R,FU H C.The mechanism of gossypol detoxification by ruminant animals[J]. The Journal of Nutrition,1962,76(3):215-218.
[30] CATER C M,LYMAN C M.Effect of bound gossypol in cottonseed meal on enzymic degradation[J]. Lipids,1970,5(9):765-769.
[31] WONG R C,NAKAGAWA Y,PERLMANN G E.Studies on the nature of the inhibition by gossypol of the transformation of pepsinogen to pepsin[J].Journal of Biological Chemistry,1972,247(5):1625-1631.
[32] FORSBERG C W,LOVELOCK L K A,KRUMHOLZ L,et al. Protease activities of rumen protozoa[J].Applied and Environmental Microbiology,1984,47(1):101-110.
[2]金红春,兰时乐,胡毅,等.棉粕发酵前后营养成分变化研究[J].饲料工业,2011,32(13):19-23.
[3] DODOU K,ANDERSON R J,SMALL D A,et al.Investigations on gossypol:past and present developments[J]. Expert Opinion on Investigational Drugs,2005,14(11):1419-1434.
[4]方琴音.饲粮不同棉籽饼用量对商品蛋鸡生产性能、蛋品质、血液生化指标的影响研究[D].硕士学位论文.雅安:四川农业大学,2004.
[5]吕云峰,王修启,赵青余,等.棉酚在饲料中安全限量及畜产品中残留研究进展[J].中国农学通报,2010,26(24):1-5.
[6] EISELE G R. A perspective on gossypol ingestion in swine[J]. Veterinary&Human Toxicology,1986,28(2):118-120.
[7] ABOU-DONIA MB.Physiological effects and metabolism of gossypol[J]. Residue Reviews,1976,61:125-160.
[8]石彩霞,侯彩云,王海荣,等.游离棉酚对绵羊瘤胃发酵和微生物区系的影响[J].中国饲料,2013(5):16-19.
[9]候彩云.棉酚对绵羊瘤胃微生物种群及纤维物质降解的影响[D].硕士学位论文.呼和浩特:内蒙古农业大学,2012.
[10] KNUTSEN H K,BARREGRD L,BIGNAMI M,et al.Presence of free gossypol in whole cottonseed[J].Efsa Journal,2017,15(7):e04850.
[11]屯妮萨·麦提赛伊迪.不同精粗比日粮、粗料型日粮添喂烟酰胺对绵羊瘤胃微生物代谢的影响[D].硕士学位论文.乌鲁木齐:新疆农业大学,2012.
[12]邓凯平,王锋,马铁伟,等.日粮中添加不同水平紫苏籽对湖羊生长性能、瘤胃发酵及养分表观消化率的影响[J].草业学报,2017,26(5):205-212.
[13]杨舒黎,王加启,胡志勇,等.日粮添加豆油和胡麻油对肉牛瘤胃发酵及主要微生物数量的影响[J].中国农业科学,2007,40(10):2316-2322.
[14]陈程.棉铃虫P450基因CPR-CYP6AE14/CYP9A12的异源共表达及其对棉酚降解作用研究[D].博士学位论文.石河子:石河子大学,2018.
[15]张龙翔,张庭芳,李玲媛.生化实验方法和技术[M].2版.北京:高等教育出版社,1997.
[16] HRISTOV A N,ROPP J K,HUNT C W. Effect of barley and its amylopectin content on ruminal fermentation and bacterial utilization of ammonia-N in vitro[J]. Animal Feed Science and Technology,2002,99(1/2/3/4):25-36.
[17] The PLOS ONE Staff. Correction:predicting in vitro rumen VFA production using CNCPS carbohydrate fractions with multiple linear models and artificial neural networks[J].PLoS One,2015,10(3):e0119740.
[18] SCHNEIDER I C,AMES ML,RASMUSSEN MA,et al.Fermentation of cottonseed and other feedstuffs in cattle rumen fluid[J].Journal of Agricultural and Food Chemistry,2002,50(8):2267-2273.
[19] FRENCH E A,BERTICS S J,ARMENTANO L E.Rumen and milk odd-and branched-chain fatty acid proportions are minimally influenced by ruminal volatile fatty acid infusions[J]. Journal of Dairy Science,2012,95(4):2015-2026.
[20]孙万成,罗毅皓,刘祥军.不同泌乳期牦牛乳中奇数与支链脂肪酸的分布[J].食品科学,2015,36(6):199-201.
[21] VLAEMINCK B,FIEVEZ V,VAN LAAR H,et al.Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates[J].Journal of Animal Physiology and Animal Nutrition,2010,88(11/12):401-411.
[22] NSEREKO V L,MORGAVI D P,RODE L M,et al.Effects of fungal enzyme preparations on hydrolysis and subsequent degradation of alfalfa hay fiber by mixed rumen microorganisms in vitro[J]. Animal Feed Science and Technology,2000,88(3/4):156-170.
[23] FESTENSTEIN G N. Cellulolytic enzymes from sheep-rumen liquor micro-organisms[J]. Biochemical Journal,1958,69(4):562-567.
[24] SUHARTATI F M,SURYAPRATAMA W.Increasing the in vitro enzymatic activity of cellulase and amylase from beef cattle rumen fluid supplemented with Moringa oleifera leaves and sulfur[J]. Pakistan Journal of Nutrition,2017,16(8):571-576.
[25]刘文杰,雒秋江,钟涛,等.添加赖氨酸对小尾寒羊瘤胃消化代谢的影响[J].中国畜牧兽医,2015,42(10):2658-2668.
[26] TRIPATHI MK,RAGHUVANSI K S K,MONDAL D,et al. Effect of Bt-cottonseed meal feeding on performance,fermentation,ciliates population and microbial hydrolytic enzymes in lamb[J].African Journal of Biotechnology,2014,13(3):510-522.
[27] LI F,ZHANG P Y,ZHANG G M,et al.Enhancement of corn stover hydrolysis with rumen fluid pretreatment at different solid contents:effect,structural changes and enzymes participation[J].International Biodeterioration&Biodegradation,2016,119:405-412.
[28] MORGAVI D P,BEAUCHEMIN K A,NSEREKO V L,et al.Resistance of feed enzymes to proteolytic inactivation by rumen microorganisms and gastrointestinal proteases[J]. Journal of Animal Science,2001,79(6):1621-1630.
[29] REISER R,FU H C.The mechanism of gossypol detoxification by ruminant animals[J]. The Journal of Nutrition,1962,76(3):215-218.
[30] CATER C M,LYMAN C M.Effect of bound gossypol in cottonseed meal on enzymic degradation[J]. Lipids,1970,5(9):765-769.
[31] WONG R C,NAKAGAWA Y,PERLMANN G E.Studies on the nature of the inhibition by gossypol of the transformation of pepsinogen to pepsin[J].Journal of Biological Chemistry,1972,247(5):1625-1631.
[32] FORSBERG C W,LOVELOCK L K A,KRUMHOLZ L,et al. Protease activities of rumen protozoa[J].Applied and Environmental Microbiology,1984,47(1):101-110.