生物活性垫料中有效菌种筛选及其在应用中影响因素研究
摘要
随着我国养殖业的飞速发展,畜禽养殖污染已经严重的破坏了自然界的生态平衡,影响了很多养殖集中区域人民的正常生活。出现农业生态环境不断恶化、动物源性食品的药物残留问题日益突出,严重的抑制了我国养殖业的可持续发展,为肉类食品埋下了安全隐患。本论文研究表明利用生物活性垫料可以有效出除生猪排泄物的异味,同时减少甚至不使用抗生素,提高了肉类的品质。
论文首先依据生猪排泄物不同成份降解需要的酶类筛选了不同微生物添加到生物活性垫料中。分别筛选了产蛋白酶菌种、产纤维素酶菌种以及耐高温菌种,然后在进行菌种的优化培养和优化组合,最后筛选到添加到生物活性垫料的菌种分别为枯草芽孢杆菌,巨大芽孢杆菌,纳豆芽孢杆菌,米曲霉,酵母,添加比例为为枯草芽孢杆菌：纳豆芽孢杆菌：巨大芽孢杆菌：米曲霉：酿酒酵母为3：1：2：1：2,总菌数为2亿cfu/g,而添加酿酒酵母可以产生酒香味,从而彻底去除生猪排泄物的异味。
在以上实验的基础上,论文还研究了不同菌种添加量对生猪排泄物降解的最佳效果,发现将枯草芽孢杆菌,纳豆芽孢杆菌,巨大芽孢杆菌,米曲霉和酿酒酵母调配到2亿cfu/g之后,按每立方米添加1 Kg的菌种到吸附剂中做成生物活性垫料,可以有效降解生猪排泄物,达到环保的要求。在南方,考虑到原材料成本问题,吸附剂选用谷壳和锯木屑,体积比为1：1的配比时,可以有效延长生物活性垫料的使用时间,达到15个月的使用时间。在猪舍改造上,并不要对传统猪舍进行推翻,而是指需要对传统猪舍进行适当改造,即可以完全满足生物活性垫料养殖的要求。论文同时探讨了消毒液对生物活性垫料是否有影响,研究发现复方戊二醛溶液和聚维酮碘溶液对生物活性垫料无明显影响,而对病原微生物有较好的抑制效果。
在小试试验的基础上,论文选择了某大型猪场进行推广试验,比较了使用生物活性垫料养殖和不使用生物活性垫料养殖。研究发现使用生物活性垫料对仔猪生长平均日增重分别提高6.93%,料肉比降低6.36%,每千克增重饲料成本降低0.29元；使用生物活性垫料对70日龄生猪生长料重比降低3.50%；每千克增重饲料成本降低0.21元。而且出栏的生猪重金属和病原微生物等都大大低于国家要求最低标准。
With the rapid development of husbandry industry in China, livestock and poultry pollution has seriously damaged the ecological balance and affected the daily life of people located around the husbandry area. Agro-ecological environment is gradually deteriorated, the issue of drug residues in food of animal origin has become increasingly prominent, which shows potential risk in meat food, all that seriously inhibiting the sustainable development of husbandry industry in China. This paper shows that the use of biological active padding can effectively remove the odor of swine excretion, even while reducing the use of antibiotics which in turn improved meat quality.
In this paper, different microorganisms, which were selected by the specific enzymes according to the components of swine excretion, were added in the biological active padding. Through screening the protease and cellulase produced bacteria and high-temperature resistant bacteria, optimized strains were cultured and combined, finally the strains which including Bacillus subtilis, Bacillus megaterium, Bacillus natto, Aspergillus oryzae, yeast were added in the padding, the ratio of added bacteria colonies of Bacillus subtilis, Bacillus subtilis natto and Bacillus megaterium is 3:1:2. Adding the appropriate amount of Aspergillus oryzae is beneficial to degrade the swine excretion, and adding the yeast can produce wine flavor which help to completely remove the odor of swine excretion.
In addition, this paper studied the different amount of bacteria, which were added to the pig excrement to get the best degradation results, and found that the Bacillus subtilis, Bacillus natto, Bacillus megaterium, Aspergillus oryzae and Saccharomyces cerevisiae were deployed to 200 million cfu /g, adding 1 Kg per cubic bacteria to adsorbent made padding with biological activity, could effectively degrade pig excrement and meet the environmental requirements. In the South of China, considering the cost of raw materials, the use of chaff and sawdust adsorbent with volume ratio of 1:1 can effectively extend the use time of the biological active padding, reaching 15months. Transformation of the piggery did not need to overthrow the transitional one, but rather referred to make appropriate transformation, which could fully meet the breeding requirements of using the biological active padding.
This paper also discussed whether the disinfectant have any impact on the biological active padding, the study found compound glutaraldehyde and povidone-iodine solution had no significant effect on the biological active padding, while they had good inhibition results on pathogenic microorganisms. Based on the result of a small pilot test, the authors selected a large farm trial for the extension test and compared the breeding results of using the biological active padding or without using the biological active padding. The study showed that the use of biological active padding could increase the average daily gain of piglets by 6.93%, decrease the feed conversion ratio by 6.36%, and decrease the feed cost of per kg weight gain by 0.29 Yuan; Besides the use of biological active padding on 70-day-old pigs could reduce the feed conversion ratio by 3.50% and the feed cost of per kilogram weight gain per kilogram by 0.21 Yuan. The amount of heavy metals and pathogenic microorganisms on slaughter pigs were much lower than the national minimum standards.
论文首先依据生猪排泄物不同成份降解需要的酶类筛选了不同微生物添加到生物活性垫料中。分别筛选了产蛋白酶菌种、产纤维素酶菌种以及耐高温菌种,然后在进行菌种的优化培养和优化组合,最后筛选到添加到生物活性垫料的菌种分别为枯草芽孢杆菌,巨大芽孢杆菌,纳豆芽孢杆菌,米曲霉,酵母,添加比例为为枯草芽孢杆菌：纳豆芽孢杆菌：巨大芽孢杆菌：米曲霉：酿酒酵母为3：1：2：1：2,总菌数为2亿cfu/g,而添加酿酒酵母可以产生酒香味,从而彻底去除生猪排泄物的异味。
在以上实验的基础上,论文还研究了不同菌种添加量对生猪排泄物降解的最佳效果,发现将枯草芽孢杆菌,纳豆芽孢杆菌,巨大芽孢杆菌,米曲霉和酿酒酵母调配到2亿cfu/g之后,按每立方米添加1 Kg的菌种到吸附剂中做成生物活性垫料,可以有效降解生猪排泄物,达到环保的要求。在南方,考虑到原材料成本问题,吸附剂选用谷壳和锯木屑,体积比为1：1的配比时,可以有效延长生物活性垫料的使用时间,达到15个月的使用时间。在猪舍改造上,并不要对传统猪舍进行推翻,而是指需要对传统猪舍进行适当改造,即可以完全满足生物活性垫料养殖的要求。论文同时探讨了消毒液对生物活性垫料是否有影响,研究发现复方戊二醛溶液和聚维酮碘溶液对生物活性垫料无明显影响,而对病原微生物有较好的抑制效果。
在小试试验的基础上,论文选择了某大型猪场进行推广试验,比较了使用生物活性垫料养殖和不使用生物活性垫料养殖。研究发现使用生物活性垫料对仔猪生长平均日增重分别提高6.93%,料肉比降低6.36%,每千克增重饲料成本降低0.29元；使用生物活性垫料对70日龄生猪生长料重比降低3.50%；每千克增重饲料成本降低0.21元。而且出栏的生猪重金属和病原微生物等都大大低于国家要求最低标准。
With the rapid development of husbandry industry in China, livestock and poultry pollution has seriously damaged the ecological balance and affected the daily life of people located around the husbandry area. Agro-ecological environment is gradually deteriorated, the issue of drug residues in food of animal origin has become increasingly prominent, which shows potential risk in meat food, all that seriously inhibiting the sustainable development of husbandry industry in China. This paper shows that the use of biological active padding can effectively remove the odor of swine excretion, even while reducing the use of antibiotics which in turn improved meat quality.
In this paper, different microorganisms, which were selected by the specific enzymes according to the components of swine excretion, were added in the biological active padding. Through screening the protease and cellulase produced bacteria and high-temperature resistant bacteria, optimized strains were cultured and combined, finally the strains which including Bacillus subtilis, Bacillus megaterium, Bacillus natto, Aspergillus oryzae, yeast were added in the padding, the ratio of added bacteria colonies of Bacillus subtilis, Bacillus subtilis natto and Bacillus megaterium is 3:1:2. Adding the appropriate amount of Aspergillus oryzae is beneficial to degrade the swine excretion, and adding the yeast can produce wine flavor which help to completely remove the odor of swine excretion.
In addition, this paper studied the different amount of bacteria, which were added to the pig excrement to get the best degradation results, and found that the Bacillus subtilis, Bacillus natto, Bacillus megaterium, Aspergillus oryzae and Saccharomyces cerevisiae were deployed to 200 million cfu /g, adding 1 Kg per cubic bacteria to adsorbent made padding with biological activity, could effectively degrade pig excrement and meet the environmental requirements. In the South of China, considering the cost of raw materials, the use of chaff and sawdust adsorbent with volume ratio of 1:1 can effectively extend the use time of the biological active padding, reaching 15months. Transformation of the piggery did not need to overthrow the transitional one, but rather referred to make appropriate transformation, which could fully meet the breeding requirements of using the biological active padding.
This paper also discussed whether the disinfectant have any impact on the biological active padding, the study found compound glutaraldehyde and povidone-iodine solution had no significant effect on the biological active padding, while they had good inhibition results on pathogenic microorganisms. Based on the result of a small pilot test, the authors selected a large farm trial for the extension test and compared the breeding results of using the biological active padding or without using the biological active padding. The study showed that the use of biological active padding could increase the average daily gain of piglets by 6.93%, decrease the feed conversion ratio by 6.36%, and decrease the feed cost of per kg weight gain by 0.29 Yuan; Besides the use of biological active padding on 70-day-old pigs could reduce the feed conversion ratio by 3.50% and the feed cost of per kilogram weight gain per kilogram by 0.21 Yuan. The amount of heavy metals and pathogenic microorganisms on slaughter pigs were much lower than the national minimum standards.
引文
[1]吉林省惠农生物科技开发有限责任公司.微生态发酵床养猪技术[J].今日畜牧兽医,2008(6)：26-28.
[2]王建华,彭君.新型发酵床养猪技术介绍[J].畜牧业,2005,34(8)：21.
[3]施光发,甘友保.土壤生物发酵床养猪技术[J].畜牧与兽医,2006,38(3)：5-39.
[4]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40-42.
[5]郑耀通,林国徐.发酵床技术在猪场清洁生产生中的应用研究[J].武夷科学,2006,22(1)：199-203.
[6]骆小俊.生物发酵床的无害化养猪技术[J].科学养殖,2009(6)：19.
[7]王远孝,李雁.猪用发酵床的研究与应用[J].家畜生态学报,2007,28(6)：139-142.
[8]王露,郭宗义.生物发酵床养猪的利与弊[J].畜禽业,2009(3)：8-10.
[9]区伟年,谭明芳.生物发酵床健康养猪技术应用与推广[J].广西畜牧兽医,2009,25(3)：162-164.
[10]倪俊娟.生物发酵舍零排放养猪技术的发展优势[J].河南畜牧兽医,2008,29(8)：13-14.
[11]毕峰.发酵床养猪法的优缺点[J].猪业科技,2008(7)：20-21.
[12]东秀珠,蔡娥英.常见细菌系统鉴定手册[M].第一版.科学出版社,北京,2001.
[13]Buchanan R. E., Giboos N. E. Bergery's manual of Determinative Bacteriology[M].8thed. Wilakins Co Baltimore.1974.
[14]赵斌,何绍江.微生物学实验[M].第一版.科学出版社,北京,2002.
[15]盘赛昆,吴文礼.一株产多糖固氮细菌的鉴定[J].淮海工学院学报,2001,10(1)：56-59.
[16]杨小蓉,宗浩.一株降解氧乐果的高效菌的分离和鉴定[J].四川师范大学学报,2001,24(4)：392-394.
[17]Sutton A. L., Kephart K. B., Verstegen M. W. A., et al. Potential for reduction of odorous compounds in swinemanure through diet modification[J]. J. Anim. Sci., 1999,77:430-439.
[18]Mitloehner F. M., Schenker M. B. Environmental exposure nd health effects from concentrated animal feeding operations[J]. Epidemiology,2007,18:309-311.
[19]柳田宏一.十着菌它利用L,亡高床式凳酵床豚含.展技街大系(畜卷环境对策)[M].震文协(E1),2002：516-522
[20]陈旭东,马秋刚,计成.芽孢杆菌制剂对仔猪生产性能的影响[J].中国饲料,2003,16：12-13.
[21]陈桂平,牛鹏霞.发酵床养猪技术及存在问题[J].现代畜牧兽医,2009(4)：10-12.
[22]王远孝.自然发酵床的研制[J].2008,40(3)：31-34.
[23]盛清凯.发酵床养猪技术的优势与推广中存在的问题[J].猪业科学,2008,(3)：80-81.
[24]国家环境保护总局自然生态保护司.全国规模化畜禽饲养业污染情况调查及防治对策[M].北京：中国环境科学出版社,2002：1-3.
[25]杜晓光.“发酵床养猪+一池三改”模式及效益分析[J].江西畜牧兽医杂志,2008,3：24-25.
[26]赵书广.对“生物发酵床”养猪的了解[J].今日养猪业,2008,(3).
[27]Gadd J. Tunnel housing of pigs in Livestock Environment Ⅳ. Fourt h International Symposium. Michigan[J]. American society of Agricultural Enginneers,1993,5:1040—1048.
[28]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40-42.
[29]毕锋.发酵床养猪的优缺点[J].科学养殖,2008(7)：20-21.
[30]王立贤.正确看待发酵床养猪[J].猪业科学,2008(9)：48.
[31]夏飚.发酵床养猪的四大好处与操作要领[J].农村养殖技术,2007(2)：17.
[32]朱永红.发酵床养猪八注意[J].畜禽养殖,2006(5)：32.
[33]张汉宝.如何科学利用发酵床养殖[J].家畜养殖,2008(18)：7.
[34]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40—42.
[35]施光发,甘友保.土壤生物发酵床养猪技术[J].畜牧兽医,2006,38(3)：59.
[36]陈合,舒国伟.产纤维素酶真菌的筛选及其产酶条件的研究[J].中国酿造,2008,180(3)：14-17.
[37]刘德海,杨玉华.纤维素酶酶活的测定方法[J].中国饲料,2002,(17)：27-28.
[38]夏服宝,邱雁临.纤维素酶活力测定条件研究[J].饲料工业,2005,16：41-44.
[39]H L Brockman. Triglyceride lipase from pOrcine pancreas.Methods Enzyme[J].1981.71:619-627.
[40]李雯,邵远志.淀粉酶活性测定方法的改进[J].植物生理学通讯,2005,45(5)：655-656.
[41]高贵,韩四平.脂肪酶活力检测方法的比较药物生物技术[J].2002.9(5)：281-284.
[42]聂国兴,王俊丽等.两种粗脂肪测定方法的比较研究[J].河南畜牧善区2001,22(7)：7-8.
[43]回瑞华,侯冬岩.荞麦中蛋白质含量的分析[J].食品科学,2004,25(10)：230-232.
[44]王十芬,周群英.环境工程微生物学[M].第3版,北京高等教育出版社,2008.
[45]和文祥,洪竖平.环境微生物学[M].北京：中国农业大学出版社,2007.
[46]刘振钦,李宏伟.发酵床养猪与水泥圈舍养猪效果比较[J].养殖技术顾问,2007,(3)：10.
[47]Kapuinen P. Deep litter system for beef cat tie housed in uninsulated barns temperatures and nutrients[J]. AgricEngng Res,2001,80(1):87-97.
[48]Deininger A, Tamrn M, Krause R, et al. Penetration resistance and waterholding capacity of dierently conditionedst raw for deep litter housing systems[J]. Agric Engng Res,2000,77 (3):335-342.
[49]陈杖榴.兽医药理学[M].北京：中国农业出版社,2001.
[50]崔中林,张彦明.现代实用动物疾病防治大全[M].北京：中国农业出版社,2001.
[51]胡元亮.兽医处方手册[M].北京：中国农业出版社,1999.
[52]费恩阁.动物传染病学[M].长春：吉林科技出版社,1995.
[2]王建华,彭君.新型发酵床养猪技术介绍[J].畜牧业,2005,34(8)：21.
[3]施光发,甘友保.土壤生物发酵床养猪技术[J].畜牧与兽医,2006,38(3)：5-39.
[4]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40-42.
[5]郑耀通,林国徐.发酵床技术在猪场清洁生产生中的应用研究[J].武夷科学,2006,22(1)：199-203.
[6]骆小俊.生物发酵床的无害化养猪技术[J].科学养殖,2009(6)：19.
[7]王远孝,李雁.猪用发酵床的研究与应用[J].家畜生态学报,2007,28(6)：139-142.
[8]王露,郭宗义.生物发酵床养猪的利与弊[J].畜禽业,2009(3)：8-10.
[9]区伟年,谭明芳.生物发酵床健康养猪技术应用与推广[J].广西畜牧兽医,2009,25(3)：162-164.
[10]倪俊娟.生物发酵舍零排放养猪技术的发展优势[J].河南畜牧兽医,2008,29(8)：13-14.
[11]毕峰.发酵床养猪法的优缺点[J].猪业科技,2008(7)：20-21.
[12]东秀珠,蔡娥英.常见细菌系统鉴定手册[M].第一版.科学出版社,北京,2001.
[13]Buchanan R. E., Giboos N. E. Bergery's manual of Determinative Bacteriology[M].8thed. Wilakins Co Baltimore.1974.
[14]赵斌,何绍江.微生物学实验[M].第一版.科学出版社,北京,2002.
[15]盘赛昆,吴文礼.一株产多糖固氮细菌的鉴定[J].淮海工学院学报,2001,10(1)：56-59.
[16]杨小蓉,宗浩.一株降解氧乐果的高效菌的分离和鉴定[J].四川师范大学学报,2001,24(4)：392-394.
[17]Sutton A. L., Kephart K. B., Verstegen M. W. A., et al. Potential for reduction of odorous compounds in swinemanure through diet modification[J]. J. Anim. Sci., 1999,77:430-439.
[18]Mitloehner F. M., Schenker M. B. Environmental exposure nd health effects from concentrated animal feeding operations[J]. Epidemiology,2007,18:309-311.
[19]柳田宏一.十着菌它利用L,亡高床式凳酵床豚含.展技街大系(畜卷环境对策)[M].震文协(E1),2002：516-522
[20]陈旭东,马秋刚,计成.芽孢杆菌制剂对仔猪生产性能的影响[J].中国饲料,2003,16：12-13.
[21]陈桂平,牛鹏霞.发酵床养猪技术及存在问题[J].现代畜牧兽医,2009(4)：10-12.
[22]王远孝.自然发酵床的研制[J].2008,40(3)：31-34.
[23]盛清凯.发酵床养猪技术的优势与推广中存在的问题[J].猪业科学,2008,(3)：80-81.
[24]国家环境保护总局自然生态保护司.全国规模化畜禽饲养业污染情况调查及防治对策[M].北京：中国环境科学出版社,2002：1-3.
[25]杜晓光.“发酵床养猪+一池三改”模式及效益分析[J].江西畜牧兽医杂志,2008,3：24-25.
[26]赵书广.对“生物发酵床”养猪的了解[J].今日养猪业,2008,(3).
[27]Gadd J. Tunnel housing of pigs in Livestock Environment Ⅳ. Fourt h International Symposium. Michigan[J]. American society of Agricultural Enginneers,1993,5:1040—1048.
[28]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40-42.
[29]毕锋.发酵床养猪的优缺点[J].科学养殖,2008(7)：20-21.
[30]王立贤.正确看待发酵床养猪[J].猪业科学,2008(9)：48.
[31]夏飚.发酵床养猪的四大好处与操作要领[J].农村养殖技术,2007(2)：17.
[32]朱永红.发酵床养猪八注意[J].畜禽养殖,2006(5)：32.
[33]张汉宝.如何科学利用发酵床养殖[J].家畜养殖,2008(18)：7.
[34]孔凡真.日本发酵床养猪技术简介[J].国际信息,2005,(2)：40—42.
[35]施光发,甘友保.土壤生物发酵床养猪技术[J].畜牧兽医,2006,38(3)：59.
[36]陈合,舒国伟.产纤维素酶真菌的筛选及其产酶条件的研究[J].中国酿造,2008,180(3)：14-17.
[37]刘德海,杨玉华.纤维素酶酶活的测定方法[J].中国饲料,2002,(17)：27-28.
[38]夏服宝,邱雁临.纤维素酶活力测定条件研究[J].饲料工业,2005,16：41-44.
[39]H L Brockman. Triglyceride lipase from pOrcine pancreas.Methods Enzyme[J].1981.71:619-627.
[40]李雯,邵远志.淀粉酶活性测定方法的改进[J].植物生理学通讯,2005,45(5)：655-656.
[41]高贵,韩四平.脂肪酶活力检测方法的比较药物生物技术[J].2002.9(5)：281-284.
[42]聂国兴,王俊丽等.两种粗脂肪测定方法的比较研究[J].河南畜牧善区2001,22(7)：7-8.
[43]回瑞华,侯冬岩.荞麦中蛋白质含量的分析[J].食品科学,2004,25(10)：230-232.
[44]王十芬,周群英.环境工程微生物学[M].第3版,北京高等教育出版社,2008.
[45]和文祥,洪竖平.环境微生物学[M].北京：中国农业大学出版社,2007.
[46]刘振钦,李宏伟.发酵床养猪与水泥圈舍养猪效果比较[J].养殖技术顾问,2007,(3)：10.
[47]Kapuinen P. Deep litter system for beef cat tie housed in uninsulated barns temperatures and nutrients[J]. AgricEngng Res,2001,80(1):87-97.
[48]Deininger A, Tamrn M, Krause R, et al. Penetration resistance and waterholding capacity of dierently conditionedst raw for deep litter housing systems[J]. Agric Engng Res,2000,77 (3):335-342.
[49]陈杖榴.兽医药理学[M].北京：中国农业出版社,2001.
[50]崔中林,张彦明.现代实用动物疾病防治大全[M].北京：中国农业出版社,2001.
[51]胡元亮.兽医处方手册[M].北京：中国农业出版社,1999.
[52]费恩阁.动物传染病学[M].长春：吉林科技出版社,1995.