黑曲霉固体发酵物的应用效果研究

摘要

为了评价由动科院畜牧微生物实验室制作的黑曲霉固体发酵物（Aspergillus niger solid culture）的使用效果，本试验采用单因子完全随机试验，设0.1%,0.2%,0.3%三个添加水平,用硫酸粘杆菌素+黄霉素和复合酶（只进行生产性能比较）作对比,选用1龄健康艾维茵肉仔鸡720只,分成6组,每组3个重复,每重复40只鸡，试验期为49天。试验结果表明：①0.1%,0.2%,0.3%A.niger SC均不同程度提高肉仔鸡0～35龄体增重,与空白对照组差异显著（P<0.05），与复合酶组和抗生素组差异不显著（P>0.05）。35～49龄各组体增重差异不显著(P>0.05)。0.2%A.niger SC显著提高了全期增重，与空白对照组差异显著（P<0.05），但与其它各处理组差异不显著（P＞0.05）。0.1%和0.3%A.niger SC提高了全期成活率，0.2%A.niger SC降低了肉仔鸡21龄成活率，但所有各组全期成活率差异不显著（P＞0.05）。

关键词：黑曲霉固体发酵物 肉仔鸡生产性能

ABSTRACT

A study was conducted to determined the effect of Aspergillus niger solid culture (A.niger SC)on growth performance, serum physiological index, intestinal microbial population, and immunological competence and digestibility of metabolizable energy and nitrogen retention of broiler..Six dietary treatments,consisting of the basal diet(control),basal diet+0.1%,0.2%or0.3% A.niger SC, basal diet+enzyme and basal +antibriotic,were fed to 600 Arbor Acres broilerchicks from 1 to 49 day of age.(enzyme is added only to compare the growth performance with the control group).The chicks were randomly assigned to 18 ages(40 chicks per cages,3cages per diet).The experimental period was 49d.Body weights,Daily feed intake , feed to gain ratio at 21,35,and 49d , serum physiological index and weight index of immune organ at 22,36,49The intestinal microbial populations at14,35,49d, proliferation of T,Blymphocyte at 28d,and digestibility at 14,20,34d were measured and determined.

The results showed that body weight gain of treatment group were improved significantly(P<0.05) at 21and 35d when compared to control broiler ,but only 0.2%A.nigerSC was significantly different from control group (P<0.05)during the 0～49d.other treatment group were not significantly different from the control group and 0.2%A.niger Sc group.All treatment group improved feed to again ratios ,decreased dead prcentage ,but not significantly different from those of control group(P>0.05). The serum physiological index were not significantly different from those of the control group(P>0.05).0.1%A.niger SC significantly improved proliferation of B lymphocyte,significantly different from that of antibrotic group(P<0.05),but not significantly different from that of the control group.0.1% A.niger SC improved weight index of thymus at 22d,spleen at 49 and bursal at 21d(P<0.05). 0.1%A.nigerSC improved crude fat digestibility at 13～１5 and 34～36,improvedAME at13～15d and 0.2%improved AME at 34～36,0.3%A.nigerSCimproved N retention at34～36d(P<0.05).Three diets containing 0.1%A.niger significantly improved Aspergillius niger ccount of rectum(P<0.05) compared with that of antibrotic group and control group.0.1%A.niger SC significantly lower the E.coli countof rectum at 14d,49d and 0.3%A.niger SC significantly lower E.coli count of rectum at 35d,49d.

As a results, 0.1%A.nigerSC was similar with those of enzyme in improved growth performance.and 0.2% A.niger SC improved the weight gain was significantly different from that of control group.A.nigerSC can regulate immunology competence ,and improved digestilibity.

Key words:Broiler Aspergillus niger solid culture production performance immunology competence digestilibity

前言

酶是由生物活性细胞产生的具有特异性催化活性的蛋白质，在生物体内具有重要的生理功能，家禽、家畜对饲料的利用,是在各种消化酶作用下将各种大分子物质降解为易被吸收、利用的小分子。但研究发现饲料中存在着一些动物难以利用的抗营养物质，原因是动物机体内缺乏消化这些营养物质的酶类。单胃动物消化道内没有分解植酸盐、纤维素、半纤维素、果胶、及其它非淀粉多糖的酶(Peeler,1972; Noy和Sklant等,1995 ;Cristofaro 等,1974 ;Hesselman 和Aman,1986)，在断奶后期的幼畜或消化道功能障碍家畜，其内源消化酶分泌不足（Jesen 和1986;计成等,1997）。同时，家畜添食的有机物中有相当一部分不能被消化，因此许多学者进行外源酶添加到饲料中以辅助动物消化的研究。研究表明，外源酶在配合饲料中的添加在提高动物的消化能力，改善饲料的利用率，消除抗营养因子，扩大可利用饲料资源，改善养殖业的生态环境方面显示出其巨大的作用。

1．酶制剂的生产方式

饲料中添加的酶制剂，绝大多数都是由微生物如酵母、曲霉菌和其它细菌来生产的。目前饲料酶制剂通过微生物发酵法生产的大体工艺是：通过选择、诱变、纯化具有分泌所需酶的微生物菌株（主要是真菌、细菌等），或通过基因重组技术（如现在的重组酶recombinant　enzyme），将产酶基因转移到成本低、能大规模发酵生产，同时又能高水平表达重组酶(recombinant　enzyme)的微生物内，进行菌种扩大培养，接种于无菌原料中，液体深层发酵或固体表层发酵，提取，浓缩，包被，载体吸附，干燥，粉碎，制成粗或浓酶制剂。

用来生产酶制剂的微生物主要采用一般公认为安全的微生物来生产。美国食品和药物管理局（FDA）规定枯草杆菌（Bacillus Subtilis），米曲霉菌（Asp.oryzae），黑曲霉菌（Asp.niger），啤酒酵母（Sacchlerevisiae）和脆壁酵母（Sacch.fnagilis）等微生物无需经鉴定可直接生产。目前饲料工业中所用微生物酶制剂很多是由曲霉菌经固体发酵生产的，固体发酵生产的酶，酶系复杂，酶不经浓缩，将发酵产品烘干后，粉碎，然后测定其活力单位，再添加填充剂，包装后成成品。这样的复合酶比单一酶更受欢迎。而液体发酵的产品一般是其中某一种酶的酶活极高，而其它酶的酶活极低。

曲霉菌是生产微生物酶制剂的主要高产菌种，其所产酶酶活性高，酶谱广，耐高温和耐较宽的pH值如植酸酶（Pallauf 等,1994;Simons 等,1990）。菌株培养基质要求不严格，易培养。曲霉属经固体发酵能产生以一种酶为主的多种酶系，在生产中有些将其固体发酵物直接低温干燥，添加一些载体就可作复合酶制剂使用，有报道表明此种复合酶制剂添加于饲料中可取得较好的效果（卢兴民,1996;刘亚力,1996;焦平林,1997）。饲料中所用酸性蛋白酶，糖化酶，纤维素酶，果胶酶及β-葡聚糖酶均可由黑曲霉菌（A.niger）发酵生产。

。。。。。。。。。

参考文献

1. 安永义.1997.肉仔鸡消化酶活发育规律机理及外源酶应用的研究.中国农业大学博士论文

2. 陈华葵樊庆笙主编.1979.微生物学.农业出版社

3. 陈陶声王大琛等主编.1987.微生物工程化学工业出版社.183-205

4. 陈远华摘译促生长抗生素与猪肠道微生物中国畜牧兽医 2002（2）：22-26

5. 冯定远王征刘玉兰.1998.β-葡聚糖酶及戊聚糖酶在猪粮中的应用.中国饲料.23:17-19

6. 冯定远张莹余石英.1997.含有木聚糖酶和β-葡聚糖酶制剂对猪粮消化性能的影响.饲料毒物与抗营养因子研究进展.汪儆主编.西安:西北大学出版社.176-180

7. 高宁国韩正康. 1997.米糠粮添加粗酶制剂对肉雏鸭生长性能、消化和代谢机能的影响.中国畜牧杂志.3:6-7

8. 管敏强毛华明.2001.提高反刍动物的饲料转化效率.兽药与饲料添加剂.3:36-38

9. 郭利川等.1994.复合酶制剂在蛋鸡配合饲料中的应用.中国饲料.3:32-33

10. 郭维烈.1996.研究微生物饲料制剂加快“白色农业”的进展.中国饲料.22:13-15

11. 郭维烈.1996.研究微生物饲料制剂加快“白色农业”的进展.中国饲料.23:13-24

12. 侯炳炎.1994 饲料用酶制剂饲料工业.15（6）：22-25

13. 计成周庆田河山.1997.断乳前（胰、小肠内容物）几种消化酶活性变化的研究.动物营养学报，9（3）：7-12

14. 蒋宗勇.1992.饲用酶制剂的生产及在动物饲养中的应用.饲料工业.13（9）：7-9

15. 焦平林等.1997.纤维素酶对肉牛及奶牛生产性能的影响.中国畜牧杂志.33(2)：43-44

16. 矍明仁主编.1999.饲料添加剂应用手册.江西科学技术出版社.133-144

17. 李新胜张春喜. 2001.加酶高梁青贮对奶牛产奶量的影响.中国奶牛.1:26-27

18. 梁运祥刘亚力.1995.固体发酵复合酶在肉鸡生产中的应用.中国饲料.16:29-30

19. 刘强汪儆等.1999.小麦粮黏滞性及小麦黏度的研究.动物营养学报.11(4):44-50

20. 刘亚力等.1996.微生物固体发酵物作复合酶饲料添加剂的研究.饲料研究.1：6-8

21. 卢兴民李广等.1996.米曲霉制剂及其不同处理方法对仔猪生产性能影响.中国饲料.10:23-24

22. 玉英.应用酶技术提高绿色饲料的营养价值.粮食与饲料工业,1995(12):22-23

23. 嘉渊译.1992.酶的测定方法.中国轻工业出版.

24. 荣俊等.1994.酶制剂在肉仔鸡饲养中的应用及某些生理生化指标的分析研究.饲料工业.15（9）：30-33

25. 沈萍主编.2000.微生物学.高等教育出版社

26. 佟建明.1999.饲用金霉素对肉仔鸡生产性能、肠道微生物和免疫机能影响研究.中国农业科学院博士学位论文

27. 王道若主编.1979.微生物学.高等医药院校教材

28. 王清吉陈伟华.大麦饲粮添加粗酶制剂对雏鸡增重、甲状腺激素水平及免疫力的影响.畜牧与兽医.1997（6）：243-245

29. 王清吉.1999.添加粗酶制剂对罗曼蛋鸡生产性能的影响.中国饲料.24-26

30. 王修启张兆敏等. 2001.加酶小麦粮对猪生产性能的影响.中国畜牧兽医.2：12-14

31. 许梓荣卢建军.2001.酶制剂GXC对生长猪稻谷型饲粮消化率的影响.浙江大学学报.27（5）：559-564

32. 杨诗兴主编.1982.饲料营养价值评定方法.甘肃人民出版社.

33. 张桂兴王云龙. 2001.饲用酶制剂的应用研究简介.国外畜牧科技.5:11-15

34. 张文举王加启等. 2001.秸杆饲料资源开发利用的研究进展.国外畜牧科技.3：15-18

35. 张先勤. 2001.寡聚糖在动物营养中的应用.兽药与饲料添加剂.5:34-36

36. 赵德英张景宏.1994.提取条件对复合酶制剂中糖化酶和纤维素酶活力测定的影响.饲料工业.15（8）：36-37

37. 周中凯.1996.饲料复合酶的研究与开发.饲料研究.7：2-3

38. 朱建津等.1994.酶制剂对仔猪消化和生长的影响.饲料工业.15（1）：28-30

39. Annison G,Choct M..1994.in:Biotechnology in the feed industry,51

40. Baidoo S K,Liu Y G,Yungbiut D.1998.Ettect of microbial enzyme supplementation on energy,amino acid digestibility and performance of pigs fed hulless barley diets.Can.J Anim.Sci.78:625-631

41. Bedford M.R.1995 Mechanism of action and poten tial environmental benefits fron the use of feed enzymes Anim.Feed.Sci.Technol.53:145-155

42. Berford,M.R.,and H.L.Classen.,1992,J.Nutr.122:560

43. Boros D,et al.1998.Site of exo-enzyme action in gastrointestinal tract of broiler chicks.Can.J Anim.Sci.78:799-602

44. Caine W R,et al.1998Effect of protease treated soybean meal on content of total soluble matter and crude protein and level of soybean trypsin inhibitors.Anim.Feed Sci.Technol.71:177-183

45. Choct M, et al.1995.Non-starch polysaccharide-degrading enzymes increase the performance for broiler chickens fed wheat and lowAPparent metabolizable energy.J Nutr.125:485-492

46. Cleophas G M L,Hartingsveldt Van.1995.Enzymes can play an important role in poultry nutrition.World Poult.11:12-13

47. Cromwell G L,Stahly T S,Coffey R D,et al.,1993 Efficacy of phytase in improving the bioavailability of phorsphous in soybean meal and corn-soybean meal diets for pigs J.Anim.Sci.71:1831-1840

48. Cromwell,G L,1980.Biological availability of phosphoruus in feedstuffs for swine.Feedstuffs 52(9):38-42

49. Eid J S G.1985.Cell wall storage carbohydrate in seeds-biochemistry of the seed “gum”and “hefmicelluloses”Adv.Botan.Res.2:125-155

50. Eming S E.1981.A study of relationships between flatus potential and carbohydrate distribution in legume seeds.J.Food Sci.46:794-803

51. Farrell D K,Martin E A.1998.Strategies to improve the nutritive value of rice bran in poultry diets.I.The addition of food enzyme to target the non-starch polysaccharide fraction in diets of xhixks and ducks gave no response.Br.Poult.Sci.39:549-554

52. Feng,P,Hunt W,Pritchard G.T,and Julien W.E. 1996 Effect of enzyme preparation on in situ and in vitro degradation and in vivo digestive characteristics of mature cool-season grass forage in beef steers J.Anim.Sci.74:1349-1357

53. Friesen K G,Nelssen J L,Goodband R D et al,1993.The effect of moist extrusion of soy products on growth performance and nutrient utilization in the early-weaned pig.J Anim.Sci.71:2099-2109

54. Gordon.R.W.et al.,1998 Influence of supplemental phytase on calcium and phosphorus utiliazation in laying hens.Poul.Sci.77:290-294

55. Hesselman,K.,and P.Aman;1986,The effect of β-glucanase on the utilization of starch and nitrogen by broiler chickens fed on barley of low and high viscosity.Anim.Feed Sci.Technol.15;83-93

56. Jackson M.1998.Comparation of 5 commercial enzymes in corn-soybean meal on broiler chick performance.Poult.Sci.77(Suppl.1):55

57. Jensen M S,Bach K E,Knudsen J et al.1998.Effect of β-glucanase supplementation on pancreatic enzyme activity and nutrient digestibility in piglets fed diets based on hulled and hulless barley varieties.Anim.Feed Sci.Technol.72:329-345

58. Johansen H N,Bach Knudsen,Wood K E et al.1997.Physico-chemical properties and the degradation of oat bran polysaccharides in the gut of pigs.J Sci.Food Agric.73:8192

59. Knudsen K E,Li B W.1991.Determination of oligosaccharides in protein-rich feedstuffs by gas-liquid chromatography. J.Agric.Food Chem.39:689-694

60. Krinke A L,.Jamroa D.1996.Effects of feed antibiotic avopramycin on organ morphology in broiler-chickens.Poult.Sci.75:705-710

61. Kuo T L,Van Middlesworth J F,Wolf W J.1988.Content of raffinose oligosaccharides and sucrose in various plant seeds.J.Agric.Food Chem.36:32-36

62. Leske K L,Jevne C J,Coon C N.1993.Effect of oligosaccharide additions on nitrogen-corrected true metabolizable energy of soy protein concentrate.Poult.Sci.72:664-668

63. Li S,Sauer W C,et al,1996a. Effect ofβ-glucanase supplementation to cereal-based diets for starter pigs on the apparent disgestibilities of dry mater ,crude protein and energy.Anim.Feed Sci.Technol.59:223-227

64. Li S,Sauer W C,et al,1996b. Effect ofβ-glucanase supplementation to hulless barley or wheat-soybean meal diets on the digestibilities of energy,protein, β-glucans and amino acids in young pig.J Anim.Sci.74:1649-1656

65. Marsmann

66. Michael R Bedford et.al ,2000,Anim Feed Sci Technol.86:1~13

67. Officer D.I.Effect of multi-enzyme supplements on the growth performance of piglets during the pre-and post-weaning periods Anim.Feed Sci.Technol.56:55-65

68. Patal M B,McGinnis J.1985.The effect of autoclaving and enzyme supplementation of guar meal on the performance of chicks and laying hens.Poult.Sci.64:1148-1156

69. Perney K M,et al,The effect of dietary phytase on growth performance and phosphorus utilization of broiler chickens.Poul Sci,72:2106-2114

70. Ronald R.Marquardt,et al.1996.Use of enzyme to improve nutrition availability in poultry feedstuffs Anim.Feed.Sci.Technol.60:321-330

71. Sebastian S et al.1996.The effects of supplemental microbial phytase on the performance and utilization of dietary calcium,phosphorus,copper,and zinc in broiler chickens fed corn-soybean diets.Poult.Sci.

72. Sebastian S,Touchburn H M G,et al.1997.Apparent digestibility of protein and amino acids in broiler chickens fed a corn-soybean diet supplemented with microbial phatase.Poult.Sci..76:1760-1769

73. Simoms P C M et al,1990.Improvement of phosphorus availability by microbial phytase in broiler and pigs.Br.J.Nutr.64:525-540

74. Smits ,C .H. M .Veldman,A.et al.1998 The inhibitiory effect of carboxymethylcellulose with high viscosity on lipid absorption in broiler chickens coincides with reduced bile salt concentration and rasised microbial numbers in the small intestine.Poult.Sci.77:1534-1539

75. Steenfeldt S,et al,1998a.Enzyme supplementation of wheat-based diet for broiler.I.Effect on growth performance and intestinal viscosity.Anim.Feed Sci.Technol.75:27-43

76. Um J S,Paik I K.1999.Effect of microbial phytase supplementation on egg production,egg shell quality,and mineral retention of laying hens fed different levels of phosphorus.Poult.Sci.78:75-79

77. Verma S V S,McNab J M.1982.Guar meal in diets for broiler chickens.Br.Poult.Sci.23:95-105

78. Vttp.and Cioccia A.M.,1993,J.Sci.Food Agri.62(4):369-374

79. White ,W.B.,et al.,1983 Viscosity of β-glucan as a factor in the enzymatic improvement of barley for chicks.Poult.Sci.62:853-862

80. White,W.B.,et al 1983,Viscosity of beta-D-glucan as a factor in the enzymatic improvement of barley for chicks.Poultry Sci.62:853-862

81. Young L G,et al.1993.Addition of microbial phytase to diets of young pigs.J Anim Sci.71:2147-2150

82. Zanella,I Sakomura, N.K et al.,1999 Effect of enzymesupplementation of diets based on corn and soybeans Poult.Sci.78:561-568

83. Zyla K and Koreleski J.1993.In-vitro and in-vivo dephosphorylation of rapesees meal by means of phytate-degrading enzymes derived from Aspergillus niger.J.Sci.Food Agric.61:1-6

84. Zyla K .David R et al.,1996The efficacy of enzymic cocktail and a fungal mycelium in dephosphorylating corn-soybean meal-based feeds fed to growing turkeys.Poult.Sci.75:381-387

85. Zyla K.Wikiera J.et al.2000.Comparison of the efficacies of a novel Aspergillus niger Mycelium with separate and combined effectiveness of phytase,acid phosphatase,and pectinase in dephosphorylation of wheat-based feeds fed to growing broilers.Poult.Sci.79:1434-1443

黑曲霉固体发酵物的应用效果研究

ABSTRACT

热门搜索

推荐论文更多

黑曲霉固体发酵物的应用效果研究

ABSTRACT

热门搜索

推荐论文 更多

推荐论文更多