高效降解棉酚菌种筛选、降解机理及固体发酵工艺研究
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摘要
棉籽粕具有较高的蛋白含量,是重要的植物源蛋白饲料原料,但其中含有棉酚及其他一些抗营养因子,这些抗营养因子对动物具有一定的毒害作用,从而极大地限制了其在畜牧生产中的饲用价值和使用量。为了提高棉籽粕的营养价值,早期研究提出了硫酸亚铁处理、氢氧化钙处理和有机溶剂浸提等一些脱毒方法,对棉籽粕的脱毒起到了重要作用,但却也存在着较多弊端。微生物发酵法是一种新型的脱毒方法,它不仅可以很好地达到脱毒的效果,同时还可以提高棉籽粕的营养价值,具有广阔的发展前景,近年来受到越来越多的重视。本课题通过高效降解棉酚菌种的选育,发酵工艺参数的优化,对微生物发酵法脱毒效果及其影响因素进行了考察分析,并采用分子生物学方法对微生物发酵脱毒的机理进行了研究。主要研究成果如下:
1、棉酚降解菌的筛选。通过采样、初筛和复筛等系统方法对棉酚降解菌株进行了筛选,得到四株棉酚降解率较高的菌株,然后采用形态学观察和分子生物学方法进行了菌种鉴定,最终确定四株菌分别为粪肠球菌(Enterococcus faecalis ),原玻璃蝇节杆菌( Arthrobacter protophormiae ),干酪乳杆菌(Lactobacillus casei),近平滑假丝酵母菌(Candida parapsilosis),并选用近平滑假丝酵母菌(C. parapsilosis KDN0118)作为主要生产菌株。
2、利用单因素法和统计学方法对C. parapsilosis KDN0118种子液体发酵的培养基成分和发酵条件进行了优化。首先利用单因素法确定了最佳碳源、最佳氮源以及初始pH、温度、转速、装液量、发酵时间的最适范围,然后采用Plackett-Burman(PB)实验设计筛选出对发酵菌量具有显著影响的关键因素酵母膏、K_2HPO_4、MgSO_4和发酵时间,最后通过Box-Behnken实验设计和SAS软件回归分析建立发酵菌量关于四个关键因素的二次多项式回归模型,并通过响应面分析法和典型性分析得出培养基最佳成分和最优发酵条件:酵母膏44.3 g/L、K_2HPO_4 7.3 g/L、MgSO_4 0.96 g/L,发酵时间为31.1 h,预测最大发酵菌量Ymax为6.55×109 cfu/mL。
3、利用单因素法和统计学方法对C. parapsilosis KDN0118固态发酵的培养基成分和发酵条件进行了优化。首先对碳源及其浓度、水分含量、初始pH、接种量、温度、翻曲次数和发酵时间进行了优化,然后利用Plackett-Burman(PB)实验设计筛选对棉酚降解率具有显著影响的关键因素Na2CO_3、FeSO_4、MgSO_4、水分含量和发酵时间,最后通过CCD实验设计和SAS软件回归分析建立棉酚降解率与关键因素之间的的二次多项式回归模型,并通过模型求解确定培养基最佳成分和最优发酵条件:Na_2CO_3 0.56 g/L,FeSO_4 72.7 mg/L,MgSO_4 0.56 g/L,水分含量58.1%和发酵时间31.6 h,预测最大棉酚降解率Ymax为60.94%。
4、以固态发酵工艺参数为基础,对微生物发酵代谢棉酚的降解机理进行了研究。运用LC-MS/MS对不同时间发酵液中的棉酚降解产物进行检测,根据MassLynx V4.1软件分析获得18种棉酚降解产物,用“元素组成”工具进行分析,推导出相应分子结构,并用Massfragment软件进行碎片分析和验证匹配,提出了C. parapsilosis KDN0118降解棉酚的代谢途径,包括与赖氨酸的结合途径及苯环的开环降解途径。
5、进行棉酚降解菌与功能菌相结合的混菌发酵工艺进行了研究。通过四株棉酚降解菌和六株功能菌的组合实验,考察了各菌株之间的相互关系,确定了棉酚降解菌与功能菌的最佳组合,并综合评价了混合菌发酵的效果以及各菌株在发酵中的作用,并通过实验验证了该混合菌组合发酵的稳定性。
Cottonseed meal is an important protein material compared with soybean meal, but the use of cottonseed meal in feed industry has been limited by the gossypol and some other anti-nutritional factors. Gossypol is a toxic polyphenolic binaphthyl dialdehyde, it can depress the growth and fertility of animal. There were several methods to remove the gossypol in cottonseed meal, including solvent extraction of free gossypol, chemical treatment with calcium hydroxide or ferrous sulfate. These methods play an important role in detoxification of cottonseed meal but many drawbacks still exist. Microbial fermentation as a new detoxification method get more and more attention, because the way is not only get the safe criteria, but also it can highly enhanced the untilization of nutrient substance. In this study, four strains with high efficiency of gossypol detoxification were screened out, the process parameters of fermentation detoxification were optimized, the factors of affecting gossypol detoxification were researched, and the mechanism of fermentation detoxification was also investigated in molecular level. The main results are as follows:
1. The screen of strains for gossypol degradation. The PDA medium containing different concentration of acetic gossypol and MRS medium with acetic gossypol were used for preliminary screening and rescreening. Cottonseed meal was then applied for practically microbial fermentation. By this means, four strains with high efficiency of gossypol detoxification were screened out from the samples of soil, rotten cottonseed meal and probiotics, then were identified as Enterococcus faecalis, Arthrobacter protophormiae, Lactobacillus casei and Candida parapsilosis by morphology and molecular biology methods. C. parapsilosis KDN0118 was then used as the main production strain.
2. The composition of Culture medium and fermentation conditions of C. parapsilosis KDN0118 seed under liquid fermentation were optimized by single-factor experiment and statistical method. In the first step, the optimal of carbon source, the nitrogen source and the range of initial pH, temperature, rotating speed, volumes of culture medium, and fermentation time were determined by single-factor experiment. As key factors of microbial growth, yeast extract, K_2HPO_4, MgSO_4 and fermentation time were then determined by Plackett-Burman design experiment. Finally, Box-Behnken design and SAS software were applied in the establishment of quadratic polynomial regression model between microbial growth and the four key factors. The optimized fermentation conditions were: yeast extract 44.3 g/L, K2HPO_4 7.3 g/L, MgSO_4 0.96 g/L, after culturing of 31.1 h, the microbial growth reached the maximum of 6.55×109cfu/mL.
3. C. parapsilosis KDN0118 was then cultured for the biodegradation of free gossypol under solidstate fermentation. Single-factor experiment and statistical methods were used in the optimization of culture medium and fermentation conditions. Carbon source and its concentration, water content, initial pH, inoculum size, temperature, stirring times and fermentation time were optimized separately. Plackett-Burman design experiment was then applied in the screening of key factors that affect the degradation of gossypol. As a result, Na_2CO_3, FeSO_4, MgSO_4, water content and fermentation time infulenced fermentation detoxification significantly. Finally, CCD design and SAS software were applied in the establishment of quadratic polynomial regression model between gossypol degradation and these key factors. Optimized culture medium and fermentation conditions obtained from model calculation were as follows: Na2CO_3 0.56 g/L, FeSO_4 72.7 mg/L, MgSO_4 0.56 g/L, moisture content 58.1%, after culturing of 31.6 h, the gossypol degradation reached the maximum of 60.94%.
4. Based on parameters of solidstate fermentation, mechanism of degradation of free gossypol in microorganism was investigated. LC-MS/MS was used in the monitoring of gossypol metabolites at intervals during fermentation. MassLynx V4.1 software analysis revealed the appearance of new products during the biodegradation process. The previously research of microbe-induced gossypol degradation was very limited. The elemental composition tool and massfragment software were used for the first time to identify the biodegradation products, as a result 18 metabolites were identified, and two degradation pathways of gossypol were proposed.
5. Mixed fermentation technology was researched by means of the combination of functional bacteria and degrading bacteria. Four strains capable of degrading gossypol and six functional strains that screened out were fermented in various combinations. The optimum combination of strains for fermentation was determined by the consideration of detoxification and nutritional value of cottonseed meal. The role of each strain in fermentation was also evaluated. Finally, stability trial of the mixed fermentation process was tested. The results showed that both the iteration and the stability were very excellent.
1、棉酚降解菌的筛选。通过采样、初筛和复筛等系统方法对棉酚降解菌株进行了筛选,得到四株棉酚降解率较高的菌株,然后采用形态学观察和分子生物学方法进行了菌种鉴定,最终确定四株菌分别为粪肠球菌(Enterococcus faecalis ),原玻璃蝇节杆菌( Arthrobacter protophormiae ),干酪乳杆菌(Lactobacillus casei),近平滑假丝酵母菌(Candida parapsilosis),并选用近平滑假丝酵母菌(C. parapsilosis KDN0118)作为主要生产菌株。
2、利用单因素法和统计学方法对C. parapsilosis KDN0118种子液体发酵的培养基成分和发酵条件进行了优化。首先利用单因素法确定了最佳碳源、最佳氮源以及初始pH、温度、转速、装液量、发酵时间的最适范围,然后采用Plackett-Burman(PB)实验设计筛选出对发酵菌量具有显著影响的关键因素酵母膏、K_2HPO_4、MgSO_4和发酵时间,最后通过Box-Behnken实验设计和SAS软件回归分析建立发酵菌量关于四个关键因素的二次多项式回归模型,并通过响应面分析法和典型性分析得出培养基最佳成分和最优发酵条件:酵母膏44.3 g/L、K_2HPO_4 7.3 g/L、MgSO_4 0.96 g/L,发酵时间为31.1 h,预测最大发酵菌量Ymax为6.55×109 cfu/mL。
3、利用单因素法和统计学方法对C. parapsilosis KDN0118固态发酵的培养基成分和发酵条件进行了优化。首先对碳源及其浓度、水分含量、初始pH、接种量、温度、翻曲次数和发酵时间进行了优化,然后利用Plackett-Burman(PB)实验设计筛选对棉酚降解率具有显著影响的关键因素Na2CO_3、FeSO_4、MgSO_4、水分含量和发酵时间,最后通过CCD实验设计和SAS软件回归分析建立棉酚降解率与关键因素之间的的二次多项式回归模型,并通过模型求解确定培养基最佳成分和最优发酵条件:Na_2CO_3 0.56 g/L,FeSO_4 72.7 mg/L,MgSO_4 0.56 g/L,水分含量58.1%和发酵时间31.6 h,预测最大棉酚降解率Ymax为60.94%。
4、以固态发酵工艺参数为基础,对微生物发酵代谢棉酚的降解机理进行了研究。运用LC-MS/MS对不同时间发酵液中的棉酚降解产物进行检测,根据MassLynx V4.1软件分析获得18种棉酚降解产物,用“元素组成”工具进行分析,推导出相应分子结构,并用Massfragment软件进行碎片分析和验证匹配,提出了C. parapsilosis KDN0118降解棉酚的代谢途径,包括与赖氨酸的结合途径及苯环的开环降解途径。
5、进行棉酚降解菌与功能菌相结合的混菌发酵工艺进行了研究。通过四株棉酚降解菌和六株功能菌的组合实验,考察了各菌株之间的相互关系,确定了棉酚降解菌与功能菌的最佳组合,并综合评价了混合菌发酵的效果以及各菌株在发酵中的作用,并通过实验验证了该混合菌组合发酵的稳定性。
Cottonseed meal is an important protein material compared with soybean meal, but the use of cottonseed meal in feed industry has been limited by the gossypol and some other anti-nutritional factors. Gossypol is a toxic polyphenolic binaphthyl dialdehyde, it can depress the growth and fertility of animal. There were several methods to remove the gossypol in cottonseed meal, including solvent extraction of free gossypol, chemical treatment with calcium hydroxide or ferrous sulfate. These methods play an important role in detoxification of cottonseed meal but many drawbacks still exist. Microbial fermentation as a new detoxification method get more and more attention, because the way is not only get the safe criteria, but also it can highly enhanced the untilization of nutrient substance. In this study, four strains with high efficiency of gossypol detoxification were screened out, the process parameters of fermentation detoxification were optimized, the factors of affecting gossypol detoxification were researched, and the mechanism of fermentation detoxification was also investigated in molecular level. The main results are as follows:
1. The screen of strains for gossypol degradation. The PDA medium containing different concentration of acetic gossypol and MRS medium with acetic gossypol were used for preliminary screening and rescreening. Cottonseed meal was then applied for practically microbial fermentation. By this means, four strains with high efficiency of gossypol detoxification were screened out from the samples of soil, rotten cottonseed meal and probiotics, then were identified as Enterococcus faecalis, Arthrobacter protophormiae, Lactobacillus casei and Candida parapsilosis by morphology and molecular biology methods. C. parapsilosis KDN0118 was then used as the main production strain.
2. The composition of Culture medium and fermentation conditions of C. parapsilosis KDN0118 seed under liquid fermentation were optimized by single-factor experiment and statistical method. In the first step, the optimal of carbon source, the nitrogen source and the range of initial pH, temperature, rotating speed, volumes of culture medium, and fermentation time were determined by single-factor experiment. As key factors of microbial growth, yeast extract, K_2HPO_4, MgSO_4 and fermentation time were then determined by Plackett-Burman design experiment. Finally, Box-Behnken design and SAS software were applied in the establishment of quadratic polynomial regression model between microbial growth and the four key factors. The optimized fermentation conditions were: yeast extract 44.3 g/L, K2HPO_4 7.3 g/L, MgSO_4 0.96 g/L, after culturing of 31.1 h, the microbial growth reached the maximum of 6.55×109cfu/mL.
3. C. parapsilosis KDN0118 was then cultured for the biodegradation of free gossypol under solidstate fermentation. Single-factor experiment and statistical methods were used in the optimization of culture medium and fermentation conditions. Carbon source and its concentration, water content, initial pH, inoculum size, temperature, stirring times and fermentation time were optimized separately. Plackett-Burman design experiment was then applied in the screening of key factors that affect the degradation of gossypol. As a result, Na_2CO_3, FeSO_4, MgSO_4, water content and fermentation time infulenced fermentation detoxification significantly. Finally, CCD design and SAS software were applied in the establishment of quadratic polynomial regression model between gossypol degradation and these key factors. Optimized culture medium and fermentation conditions obtained from model calculation were as follows: Na2CO_3 0.56 g/L, FeSO_4 72.7 mg/L, MgSO_4 0.56 g/L, moisture content 58.1%, after culturing of 31.6 h, the gossypol degradation reached the maximum of 60.94%.
4. Based on parameters of solidstate fermentation, mechanism of degradation of free gossypol in microorganism was investigated. LC-MS/MS was used in the monitoring of gossypol metabolites at intervals during fermentation. MassLynx V4.1 software analysis revealed the appearance of new products during the biodegradation process. The previously research of microbe-induced gossypol degradation was very limited. The elemental composition tool and massfragment software were used for the first time to identify the biodegradation products, as a result 18 metabolites were identified, and two degradation pathways of gossypol were proposed.
5. Mixed fermentation technology was researched by means of the combination of functional bacteria and degrading bacteria. Four strains capable of degrading gossypol and six functional strains that screened out were fermented in various combinations. The optimum combination of strains for fermentation was determined by the consideration of detoxification and nutritional value of cottonseed meal. The role of each strain in fermentation was also evaluated. Finally, stability trial of the mixed fermentation process was tested. The results showed that both the iteration and the stability were very excellent.
引文
[1]郭维烈,郭庆华.新型发酵蛋白饲料[M].科学技术文献出版社, 2005.
[2]林文辉.开源节流是解决饲料蛋白源短缺的根本途径[J].北方牧业. 2007, (022): 8-8.
[3]施安辉,单宝龙,贾朋辉,等.国内蛋白质饲料资源开发利用的现状及前景[J].饲料博览. 2006, 6: 40-43.
[4]沙玉圣.当前饲料工业和养殖业生产形势分析[J].中国饲料. 2007, (020): 2-4.
[5]石学刚,王斯佳,李发弟,等.动物性蛋白饲料原料开发及应用现状[J].中国畜牧杂志. 2007, 43(020): 46-50.
[6]张日俊.体外消化――提高蛋白利用率和降低饲料成本的必然选择[J].北方牧业. 2007, (022): 7-7.
[7]周围.蛋白原料价格上涨企业如何降低饲料成本[J].北方牧业. 2007, (022): 7-7.
[8]李爱科,郝淑红,伍松陵.植物蛋白质饲料资源开发利用新技术研究进展[J].饲料与畜牧. 2006, 10: 5-9.
[9]王树栋.棉籽饼粕综述[J].中国奶牛. 1999, (002): 22-23.
[10]沈慧乐等.实用家禽营养[J]. 2002: 186-187.
[10]张继东,王志祥,丁景华,等.棉籽饼粕中天然抗营养因子的危害机理及消除措施[J].畜牧与饲料科学. 2006, 27(003): 53-55.
[11]郭香墨,范术丽,王红梅,等.我国棉花育种技术的创新与成就[J].棉花学报. 2007, 19(5): 323-330.
[12] Pandey S, Thejappa N. Study on relationship between oil, protein, and gossypol in cottonseed kernels[J]. Journal of the American Oil Chemists' Society. 1975, 52(8): 312-315.
[13]中国农业科学院畜牧研究所.棉籽饼(粕)营养成分及有毒有害物质含量[M].北京:中国农业科学院畜牧研究所,1990:1-40.
[14]贾晓红, Wedegaertner T.充分利用棉籽粕[J].中国畜牧兽医. 1982, 6: 11-16.
[15]周伟良,尚建波.棉籽饼粕的价值及游离棉酚检测方法[J].江西饲料. 1996, (003): 14-15.
[16]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.
[17]沈维华,马承融.棉籽制油工艺对棉籽蛋白品质的影响[J].中国油脂. 1988, 6: 27-31.
[18] Waldroup P, H. KJ. Nutrient composition of cottonseed meal[J]. Feedstuffs. 2002, 74(45): 11-12.
[19]张贤亮,邹焰峰,孙秋亮.奎屯,石河子垦区部分团场棉籽饼棉酚含量的测定[J].新疆农垦科技. 1990, 4(7).
[20]马春晖,万英.南疆地区部分县,团场棉籽饼棉酚含量的测定[J].塔里木农垦大学学报. 1994, 6(001): 30-32.
[21]李德发.中国饲料大全[M].北京:中国农业出版社, 2001.
[22]王安平.棉粕和棉籽营养成分与棉酚含量测定及其对奶牛产奶量和乳品质的影响[J].湖南农业大学, 2009.
[23] Yang W, Xiang S. Separation of Gossypol Enantiomers byμBondapak C18 Column[M]. Sepu,1995.
[24] Brocas C, Rivera R, Paula-Lopes F, et al. Deleterious actions of gossypol on bovine spermatozoa, oocytes, and embryos[J]. Biology of Reproduction. 1997, 57(4): 901.
[25] Chenoweth P, Chase C. Characterization of gossypol-induced sperm abnormalities in bulls[J]. Theriogenology. 2000, 53(5): 1193-1203.
[26] Yildirim-Aksoy M, Lim C, Wan P, et al. Effect of natural free gossypol and gossypol-acetic acid on growth performance and resistance of channel catfish (Ictalurus punctatus) to Edwardsiella ictaluri challenge[J]. Aquaculture Nutrition. 2004, 10(3): 153-165.
[27] Randel R, Chase Jr C, Wyse S. Effects of gossypol and cottonseed products on reproduction of mammals[J]. Journal of Animal Science. 1992, 70(5): 1628.
[28] Risco CA, Holmberg CA, Kutches A. Effect of Graded Concentrations of Gossypol on Calf Performance: Toxicological and Pathological Considerations1[J]. Journal of dairy science. 1992, 75(10): 2787-2798.
[29] Lordelo M, Davis A, Calhoun M, et al. Relative toxicity of gossypol enantiomers in broilers[J]. Poultry science. 2005, 84(9): 1376.
[30] Blom J, Lee K, Rinchard J, et al. Reproductive efficiency and maternal-offspring transfer of gossypol in rainbow trout (Oncorhynchus mykiss) fed diets containing cottonseed meal[J]. Journal of Animal Science. 2001, 79(6): 1533.
[31] Bender HS, Saunders GK, Misra HP. A histopathologic study of the effects of gossypol on the female rat[J]. Contraception. 1988, 38(5): 585-592.
[32] Bozek SA, Jensen D, Tone J. Scanning electron microscopic study of spermatozoa from gossypol-treated rats[J]. Cell and Tissue Research. 1981, 219(3): 659-663.
[33] Oko R, Hrudka F. Segmental aplasia of the mitochondrial sheath and sequelae induced by gossypol in rat spermatozoa[J]. Biology of Reproduction. 1982, 26(1): 183.
[34] Hoffer AP. Ultrastructural, biochemical and endocrine studies on the effects of gossypol and its isomeric derivatives on the male reproductive tract[J]. Gossypol, a potential contraceptive for men, Plenum Press, New York. 1985: 143?186.
[35] Panigrahi S, Plumb V, Machin D. Effects of dietary cottonseed meal, with and without iron treatment, on laying hens[J]. British poultry science. 1989, 30(3): 641.
[36] Panigrahi S, Hammonds T. Egg discolouration effects of including screw-press cottonseed meal in laying hen diets and their prevention[J]. British poultry science. 1990, 31(1): 107-120.
[37] Ciereszko A, Dabrowski K. In vitro effect of gossypol acetate on yellow perch (Perca flavescens) spermatozoa[J]. Aquatic toxicology. 2000, 49(3): 181-187.
[38] Dabrowski K, Lee KJ, Rinchard J, et al. Gossypol isomers bind specifically to blood plasmaproteins and spermatozoa of rainbow trout fed diets containing cottonseed meal[J]. Biochimica et Biophysica Acta (BBA)-General Subjects. 2001, 1525(1-2): 37-42.
[39] Rinchard J, Lee K, Czesny S, et al. Effect of feeding cottonseed meal-containing diets to broodstock rainbow trout and their impact on the growth of their progenies[J]. Aquaculture. 2003, 227(1-4): 77-87.
[40] Lee KJ, Rinchard J, Dabrowski K, et al. Long-term effects of dietary cottonseed meal on growth and reproductive performance of rainbow trout: Three-year study[J]. Animal feed science and technology. 2006, 126(1-2): 93-106.
[41]张庆朝,王化春.棉酚对畜禽的毒害作用[J].中国兽医科技. 1994, 24(010): 17-18.
[42]卢智文.日粮中棉酚含量计算方法及安全限量[J].中国饲料. 1996, 24: 33-34.
[43] Longmore J. Cottonseed oil: Its coloring matter mucilage, and description of a new method of recovering the loss occurring in the refining process[J]. J. Soc. Chem. Ind. 1886, 5: 200.
[44] Berardi L, Goldblatt C, Liener I. Gossypol[J]. Toxic constituents of plant foodstuffs. 1980: 183-237.
[45] Campbell K, Morris R, Adams R. The Structure of Gossypol. I[J]. Journal of the American Chemical Society. 1937, 59(9): 1723-1728.
[46] Hron R, Koltun S, Pominski J, et al. The potential commercial aspects of gossypol[J]. Journal of the American Oil Chemists' Society. 1987, 64(9): 1315-1319.
[47] Marin A, Shlyapnikov Y, Mametov E, et al. Retardation of polymer oxidation by natural antioxidant gossypol. Part 1: kinetics of oxygen consumption[J]. Polymer degradation and stability. 1992, 35(2): 141-146.
[48]王飞,米海峰.棉籽饼粕中棉酚对动物机体的毒害作用及其预防措施[J].粮食与饲料工业. 2003, (008): 27-28.
[49]白玉萍,于树森,都徐亮.挤压膨化机在棉籽,米糠油料加工中的应用[J].中国油脂. 2004, 29(012): 36-37.
[50] Rhee K, Ziprin Y, Calhoun M. Antioxidative effects of cottonseed meals as evaluated in cooked meat[J]. Meat science. 2001, 58(2): 117-123.
[51] Lusas E, Jividen G. Glandless cottonseed: A review of the first 25 years of processing and utilization research[J]. Journal of the American Oil Chemists' Society. 1987, 64(6): 839-854.
[52]黄祖德.混合溶剂一次浸出棉仁脱酚[J].中国油脂. 1994, 19(002): 25-27.
[53]刘大川,阎杰.溶剂法浸出棉籽油和脱除棉酚的工艺研究[J].中国油脂. 1999, 24(004): 6-8.
[54]戴卫东,李泉,钱礼华,等丙酮,己烷与水混合溶剂脱除棉酚的实验研究[J].化学与生物工程. 2004, 21(005): 32-33.
[55]祝强,刘进才,唐金泉.一种新的棉籽脱酚生产工艺[J].中国油脂. 2004, 29(001): 68-70.
[56] Rahma E, Raoms N. Gossypol removal and functional properties of protein produced byextraction of glanded cottonseed with different solvents[J]. Journal of Food Science. 1984, 49(4): 1057-1060.
[57] Johnson L, Farnsworth J, Sadek N, et al. Pilot plant studies on extracting cottonseed with methylene chloride[J]. Journal of the American Oil Chemists' Society. 1986, 63(5): 647-652.
[58] Abraham G, Decossas K, Hron R, et al. Process engineering economic evaluation of the ethanol extraction of cottonseed: preliminary analysis[J]. Journal of the American Oil Chemists' Society. 1991, 68(6): 418-421.
[59] Lusas EW, Watkins LR, Koseoglu S. Isopropyl alcohol to be tested as solvent[J]. International news on fats, oils and related materials (USA). 1991.
[60] Cherry JP, Gray MS. Methylene chloride extraction of gossypol from cottonseed products[J]. Journal of Food Science. 1981, 46(6): 1726-1733.
[61]乔国平,王立.棉籽饼粕脱毒方法研究进展[J].粮食与油脂. 2002, (003): 27-29.
[62] Kamga R, Kayem GJ, Rouxhet PG. Adsorption of gossypol from cottonseed oil on oxides[J]. Journal of colloid and interface science. 2000, 232(1): 198-206.
[63]石秀侠,程茂基,蔡克周,等.棉籽饼有毒物质及其脱毒方法研究进展[J].饲料博览. 2005, (006): 8-10.
[64]阎轶沽,宋维平,张建云.棉籽饼粕在畜禽中的应用及棉酚的脱毒方法研究[J].饲料工业. 2005, 26(003): 47-50.
[65] Nagalakshmi D, Sastry V, Agrawal D. Detoxification of undecorticated cottonseed meal by various physical and chemical methods[J]. Animal Nutrition and Feed Technology. 2002, 2(2): 117-126.
[66] Nagalakshmi D, Sastry V, Pawde A. Rumen fermentation patterns and nutrient digestion in lambs fed cottonseed meal supplemental diets[J]. Animal feed science and technology. 2003, 103(1-4): 1-14.
[67] Tabatabai F, Golian A, Salarmoeini M. Determination and detoxification methods of cottonseed meal gossypol for broiler chicken rations[J]. Agricultural Sciences and Technology. 2002.
[68]李延云,聂宇燕.棉籽饼粕工业化脱毒生产技术[J].饲料工业. 1999, 20(002): 11-12.
[69]张跃顺.棉籽饼的脱毒方法及其效果[J].山东畜牧兽医. 2004, (001): 13-13.
[70] Barraza M, Coppock C, Brooks K, et al. Iron Sulfate and Feed Pelleting to Detoxify Free Gossypol in Cottonseed Diets for Dairy Cattle1[J]. Journal of dairy science. 1991, 74(10): 3457-3467.
[71]仲明,高庆华.棉籽粕几种脱毒方法效果的比较[J].中国饲料. 1998, (006): 31-32.
[72]张嗣炯.棉籽饼粕脱毒工艺的研究[J].中国粮油学报. 1995, 10(004): 55-58.
[73]高金宝.棉籽料坯高碱水蒸炒[J].中国油脂. 1998, 23(001): 55-56.
[74]戴卫东,卢伯南,钱礼华,等.热碱法脱除游离棉酚的实验研究[J].应用化工. 2004, 33(006): 57-59.
[75]赵兴华,何欣,董强,等.植物毒素的微生物降解与利用研究进展[J].广西农业生物科学. 2006, 25(B09): 172-177.
[76] El-Saidy DMSD, Gaber MM. Use of cottonseed meal supplemented with iron for detoxification of gossypol as a total replacement of fish meal in Nile tilapia, Oreochromis niloticus (L.) diets[J]. Aquaculture Research. 2004, 35(9): 859-865.
[77]杨晓斌,林庆生.微生物饲料添加剂的研究进展[J].氨基酸和生物资源. 2003, 25(002): 10-13.
[78]徐姗楠,邱宏端.微生物发酵生产蛋白饲料的研究进展[J].福州大学学报:自然科学版. 2002, (0z1): 709-713.
[79] Weng XY, Sun JY. Biodegradation of free gossypol by a new strain of Candida tropicalis under solid state fermentation: Effects of fermentation parameters[J]. Process Biochemistry. 2006, 41(7): 1663-1668.
[80] Weng XY, Sun JY. Kinetics of biodegradation of free gossypol by Candida tropicalis in solid-state fermentation[J]. Biochemical Engineering Journal. 2006, 32(3): 226-232.
[81] Reiser R, Fu HC. The mechanism of gossypol detoxification by ruminant animals[J]. The Journal of Nutrition. 1962, 76(2): 215.
[82]钟英长,吴玲娟.棉籽饼的微生物脱毒法研究[J].饲料工业. 1989, (008): 4-8.
[83] Rajarathnam S, Shashirekha M, Bano Z. Biodegradation of gossypol by the white oyster mushroom, Pleurotus florida, during culturing on rice straw growth substrate, supplemented with cottonseed powder[J]. World Journal of Microbiology and Biotechnology. 2001, 17(3): 221-227.
[84]钟英长,吴玲娟.利用微生物将棉籽中游离棉酚脱毒的研究[J].中山大学学报(自然科学版). 1989, 3.
[85]施安辉,黄玉梅.微生物脱毒强化棉籽饼的研究与应用[J].粮食与饲料工业. 1997, (007): 22-23.
[86]孙建义,许梓荣.利用假丝酵母进行棉籽饼脱毒的研究[J].中国粮油学报. 1995, 10(001): 61-64.
[87]顾赛红,孙建义,李卫芬.黑曲霉PES固体发酵对棉籽粕营养价值的影响[J]. JOURNAL OF THE CHINESE CEREALS AND OILS ASSOCIATION. 2003, 18(1).
[88]张文举,许梓荣,孙建义,等.假丝酵母ZD-3与黑曲霉ZD-8复合固体发酵对棉籽饼脱毒及营养价值的影响研究[J].中国粮油学报. 2006, 21(006): 129-135.
[89]院江,孙新文,丁宁,等.微生物发酵对棉籽壳营养成分及游离棉酚的影响[J].石河子大学学报. 2006, 24(003): 299-301.
[90]张庆华,赵新海,钟丽娟,等.三菌株协同固态发酵对棉粕脱毒效果及其生物活性的影响[J].饲料工业. 2007, 28(018): 37-38.
[91] Yude H. Effect of degossypolized cottonseed meal by microbial method on the performance of layers[J]. Chinese Journal of Animal Science (China). 1994.
[92] Marquie C, Aymard C, Cuq JL, et al. Biodegradable packaging made from cottonseed flour:formation and improvement by chemical treatments with gossypol, formaldehyde, and glutaraldehyde[J]. Journal of Agricultural and Food Chemistry. 1995, 43(10): 2762-2767.
[93] Shashirekha M, Rajarathnam S, Bano Z. Enhancement of bioconversion efficiency and chemistry of the mushroom, Pleurotus sajor-caju (Berk and Br.) Sacc. produced on spent rice straw substrate, supplemented with oil seed cakes[J]. Food chemistry. 2002, 76(1): 27-31.
[94] Zhang WJ, Xu ZR, Zhao SH, et al. Optimization of process parameters for reduction of gossypol levels in cottonseed meal by Candida tropicalis ZD-3 during solid substrate fermentation[J]. Toxicon. 2006, 48(2): 221-226.
[95] Zhang WJ, Xu ZR, Zhao SH, et al. Development of a microbial fermentation process for detoxification of gossypol in cottonseed meal[J]. Animal feed science and technology. 2007, 135(1-2): 176-186.
[96]吴小月,陈金湘.利用微生物降解棉仁饼粕中游离棉酚的研究[J].中国农业科学. 1989, 22(2): 82-86.
[97]杨景芝,孙衍华.棉酚脱毒微生物的筛选及其脱毒效果的研究[J].山东农业大学学报:自然科学版. 1999, 30(001): 26-30.
[98]何涛,滕达,张海军,等.微生物降解棉酚的响应面分析优化[J].食品与发酵工业. 2008, 34(004): 71-74.
[99]吴伟伟,许赣荣.复合微生物固态发酵对棉籽饼粕脱毒及营养的影响[J]. INDUSTRIAL MICROBIOLOGY. 2010, 40(3).
[100] Smith F. Determination of gossypol in leaves and flower buds of Gossypium[J]. Journal of the American Oil Chemists' Society. 1967, 44(4): 267-269.
[101]宁开桂.实用饲料分析手册[M]..中国农业科技出版社, 1993.
[102] Lee KJ, Dabrowski K. High-performance liquid chromatographic determination of gossypol and gossypolone enantiomers in fish tissues using simultaneous electrochemical and ultraviolet detectors[J]. Journal of Chromatography B. 2002, 779(2): 313-319.
[103]李信,刘云,计林贞.侧孢霉利用玉米秸秆固体发酵产生木质纤维素酶的研究[J].核农学报. 2000, 14(2): 99-103.
[104]张文举.高效降解棉酚菌种的选育及棉籽饼粕生物发酵的研究[J].浙江大学, 2006.
[105]院江,孙新文,蔡铁查.棉籽壳发酵前后游离棉酚及营养成分变化的实验[J].饲料研究. 2006, (004): 3-4.
[106]孙中涛.苹果渣与棉粕固态发酵生产NSP酶的研究[D].山东农业大学, 2008.
[107]布坎南RE.伯杰氏细菌鉴定手册[M].北京:科学出版社, 1988.
[108]贾晓锋.固态发酵对棉籽粕棉酚脱毒及蛋白质降解的影响[J].西北农林科技大学, 2008.
[109] Jia Xiaofeng NA. Effect of solid-state fermentation on gossypol detoxification and protein degradation in cottonseed meal[J]. Journal of Northwest Agriculture and Forestry University. 2009, 37.
[110]汤江武,孙宏,姚晓红,等.响应面法与基于神经网络的遗传算法在优化棉粕固态发酵中外源添加物用量上的应用[J].浙江大学学报(农业与生命科学版). 2011, 37(1): 97-102.
[111] Murray BE, Singh KV, Markowitz SM, et al. Evidence for clonal spread of a single strain ofβ-lactamase-producing Enterococcus (Streptococcus) faecalis to six hospitals in five states[J]. Journal of Infectious Diseases. 1991, 163(4): 780.
[112]刘凤强,姬生瑞.检出160株肠球菌的分布及耐药性分析[J].中国热带医学. 2005, 5(009): 1879-1879.
[113] Murray BE. The life and times of the Enterococcus[J]. Clinical Microbiology Reviews. 1990, 3(1): 46.
[114]段永兰.益生菌与动物饲料[J].郑州牧业工程高等专科学校学报. 2007, 27(002): 31-33.
[115] Dimov SG. A novel bacteriocin-like substance produced by Enterococcus faecium 3587[J]. Current Microbiology. 2007, 55(4): 323-327.
[116] Strompfova V, Laukova A, Ouwehand AC. Selection of enterococci for potential canine probiotic additives[J]. Veterinary microbiology. 2004, 100(1-2): 107-114.
[117] Settanni L, Valmorri S, Suzzi G, et al. The role of environmental factors and medium composition on bacteriocin-like inhibitory substances (BLIS) production by Enterococcus mundtii strains[J]. Food microbiology. 2008, 25(5): 722-728.
[118] Callewaert R, Hugas M, Vuyst LD. Competitiveness and bacteriocin production of Enterococci in the production of Spanish-style dry fermented sausages[J]. International Journal of Food Microbiology. 2000, 57(1-2): 33-42.
[119] Moon GS, Kang CH, Pyun YR, et al. Isolation, identification, and characterization of a bacteriocin-producing Enterococcus sp. from kimchi and its application to kimchi fermentation[J]. Journal of microbiology and biotechnology. 2004, 14(5): 924-931.
[120] Kim J, Oh M, Rhee Y, et al. Selection and physico-chemical characteristics of lactic acid bacteria which had cholesterol lowering activities[J]. The Korean Society of Agricultural Chemistry and Biotechnology. 1999.
[121] Hlivak P, Odraska J, Ferencik M, et al. One-year application of probiotic strain Enterococcus faecium M-74 decreases serum cholesterol levels[J]. Bratisl Lek Listy. 2005, 106(2): 67-72.
[122] Martínez?Bueno M, Valdivia E, Gálvez A, et al. Analysis of the gene cluster involved in production and immunity of the peptide antibiotic AS?48 in Enterococcus faecalis[J]. Molecular microbiology. 1998, 27(2): 347-358.
[123] Benyacoub J, Czarnecki-Maulden GL, Cavadini C, et al. Supplementation of food with Enterococcus faecium (SF68) stimulates immune functions in young dogs[J]. The Journal of Nutrition. 2003, 133(4): 1158.
[124] Hollis A, Wilkins P, Palmer J, et al. Bacteremia in equine neonatal diarrhea: a retrospectivestudy [J]. Journal of Veterinary Internal Medicine. 2008, 22(5): 1203-1209.
[125] Knorr R, Cavadini C, Benyacoub J, et al. Probiotic Enterococci For Improved Immunity [M]. EP Patent, 2010.
[126] Tanaka NG, Sakamoto N, Inoue K , et al. Antitumor effects of an antiangiogenic polysaccharide from an Arthrobacter species with or without a steroid[J]. Cancer research. 1989, 49(23): 6727.
[127] Xiong G, Maser E. Regulation of the steroid-inducible 3α-hydroxysteroid dehydrogenase /carbonyl reductase gene in Comamonas testosteroni[J]. Journal of Biological Chemistry. 2001, 276(13): 9961.
[128] Metchnikoff E, Metchnikoff II, Mitchell SPC. The prolongation of life: optimistic studies[m]. Springer Publishing Company, 2004.
[129]孔祥武,钮跃贞,陈圣洁,等.微生态制剂的临床应用[J].中华实用医药杂志. 2003, 3(6).
[130] Fioramonti J, Theodorou V, Bueno L. Probiotics: what are they? What are their effects on gut physiology?[J]. Best Practice & Research Clinical Gastroenterology. 2003, 17(5): 711-724.
[131] Alonso L, Cuesta E, Gilliland S. Production of Free Conjugated Linoleic Acid by Lactobacillus acidophilus and Lactobacillus casei of Human Intestinal Origin1[J]. Journal of dairy science. 2003, 86(6): 1941-1946.
[132] Furushiro M, Sawada H, Hirai K, et al. Blood Pressure-lowering Effect of Extract from Lactobacillus casei in Spontaneously Hypertensive Rats (SHR)(Microbiology & Fermentation Industry)[J]. Agricultural and biological chemistry. 1990, 54(9): 2193-2198.
[133] Liong M, Shah N. Acid and bile tolerance and cholesterol removal ability of lactobacilli strains[J]. Journal of dairy science. 2005, 88(1): 55-66.
[134] Aso Y, Akaza H, Kotake T , et al. Preventive effect of a Lactobacillus casei preparation on the recurrence of superficial bladder cancer in a double-blind trial[J]. European urology. 1995, 27(2): 104-109.
[135] Reid G. The scientific basis for probiotic strains of Lactobacillus[J]. Applied and environmental Microbiology. 1999, 65(9): 3763.
[136] Zhang R, Xu Y, Sun Y , et al. Construction of an enzyme-coupled system consisting of (R)-and (S)-specific carbonyl reductases for one-step preparation of (S)-1-phenyl-1, 2-ethanediol[J]. Wei sheng wu xue bao= Acta microbiologica Sinica. 2009, 49(2): 204.
[137]褚以文.微生物培养基优化方法及其OPTI优化软件[J].国外医药:抗生素分册. 1999, 20(002): 58-60.
[138] Ahuja S, Ferreira G, Moreira A. Application of Plackett?Burman design and response surface methodology to achieve exponential growth for aggregated shipworm bacterium[J]. Biotechnology and bioengineering. 2004, 85(6): 666-675.
[139] Loukas YL. A Plackett-Burnam screening design directs the efficient formulation ofmulticomponent DRV liposomes[J]. Journal of pharmaceutical and biomedical analysis. 2001, 26(2): 255-263.
[140] Hanrahan G, Lu K. Application of factorial and response surface methodology in modern experimental design and optimization[J]. Critical Reviews in Analytical Chemistry. 2006, 36(3-4): 141-151.
[141]杨承剑,黄兴国,李伟,等. Plackett-Burman设计在益生菌生长主要影响因子筛选中的应用[J].饲料工业. 2007, 28(016): 31-33.
[142] Vindevogel J, Sandra P. Resolution optimization in micellar electrokinetic chromatography: use of Plackett-Burman statistical design for the analysis of testosterone esters[J]. Analytical Chemistry. 1991, 63(15): 1530-1536.
[143] Ghanem NB, Yusef HH, Mahrouse HK. Production of Aspergillus terreus xylanase in solid-state cultures: application of the Plackett-Burman experimental design to evaluate nutritional requirements[J]. Bioresource technology. 2000, 73(2): 113-121.
[144] Rao P, Divakar S. Lipase catalyzed esterification of [alpha]-terpineol with various organic acids: application of the Plackett-Burman design[J]. Process Biochemistry. 2001, 36(11): 1125-1128.
[145] Khandeparkar R, Bhosle N. Isolation, purification and characterization of the xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation[J]. Enzyme and microbial technology. 2006, 39(4): 732-742.
[146] Walker GM. Metals in yeast fermentation processes[J]. Advances in applied microbiology. 2004, 54: 197-229.
[147] Puri M, Banerjee A, Banerjee U. Optimization of process parameters for the production of naringinase by Aspergillus niger MTCC 1344[J]. Process Biochemistry. 2005, 40(1): 195-201.
[148] Prawirodigdo S, Gannon N, Van Barneveld R , et al. Assessment of apparent ileal digestibility of amino acids and nitrogen in cottonseed and soyabean meals fed to pigs determined using ileal dissection under halothane anaesthesia or following carbon dioxide-stunning[J]. British Journal of Nutrition. 1998, 80(02): 183-191.
[149] Knabe D, LaRue D, Gregg E , et al. Apparent digestibility of nitrogen and amino acids in protein feedstuffs by growing pigs[J]. Journal of Animal Science. 1989, 67(2): 441.
[150]杨继良,周大云,杨伟华,等.高效降解棉酚菌种的筛选及棉饼脱毒参数的研究[J].棉花学报. 2002, 12(5): 225-229.
[151]肖永友,姜玉,王沦娟,蔡发国.高效降解棉酚菌株的筛选及其发酵技术研究[J].广东饲料. 2008, 3:34-36.
[152]朱德伟,刘志鹏,陆建,等.高效降解棉酚菌种的筛选及棉粕发酵脱毒工艺研究[J].中国油脂. 2010 ,35 (2): 24-28.
[153]聂蓬勃,汤江武,梁运祥.棉粕脱毒菌株的筛选及发酵条件的研究[J].浙江农业科学. 2009, 1:120-122.
[154] Pettit R K. Mixed fermentation for natural product drug discovery[J]. Applied microbiology and biotechnology. 2009, 83(1): 19-25.
[155] Schneider IC, 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: 2267-2273.
[156] Abou-Donia MB, Lyman CM, Dieckert JW. Metabolic fate of gossypol: the metabolism of 14C-gossypol in rats[J]. Lipids. 1970,5: 938-946.
[157] Abou-Donia MB, Dieckert JW. Metabolic fate of gossypol: the metabolism of [14C]gossypol in swine[J]. Toxicology and Applied Pharmacology. 1975,31: 32-46.
[158] Bajpai M, Gupta G, Setty BS. Changes in carbohydrate metabolism of testicular germ cells during meiosis in the rat[J]. European Journal of Endocrinology. 1998, 138: 322-327.
[2]林文辉.开源节流是解决饲料蛋白源短缺的根本途径[J].北方牧业. 2007, (022): 8-8.
[3]施安辉,单宝龙,贾朋辉,等.国内蛋白质饲料资源开发利用的现状及前景[J].饲料博览. 2006, 6: 40-43.
[4]沙玉圣.当前饲料工业和养殖业生产形势分析[J].中国饲料. 2007, (020): 2-4.
[5]石学刚,王斯佳,李发弟,等.动物性蛋白饲料原料开发及应用现状[J].中国畜牧杂志. 2007, 43(020): 46-50.
[6]张日俊.体外消化――提高蛋白利用率和降低饲料成本的必然选择[J].北方牧业. 2007, (022): 7-7.
[7]周围.蛋白原料价格上涨企业如何降低饲料成本[J].北方牧业. 2007, (022): 7-7.
[8]李爱科,郝淑红,伍松陵.植物蛋白质饲料资源开发利用新技术研究进展[J].饲料与畜牧. 2006, 10: 5-9.
[9]王树栋.棉籽饼粕综述[J].中国奶牛. 1999, (002): 22-23.
[10]沈慧乐等.实用家禽营养[J]. 2002: 186-187.
[10]张继东,王志祥,丁景华,等.棉籽饼粕中天然抗营养因子的危害机理及消除措施[J].畜牧与饲料科学. 2006, 27(003): 53-55.
[11]郭香墨,范术丽,王红梅,等.我国棉花育种技术的创新与成就[J].棉花学报. 2007, 19(5): 323-330.
[12] Pandey S, Thejappa N. Study on relationship between oil, protein, and gossypol in cottonseed kernels[J]. Journal of the American Oil Chemists' Society. 1975, 52(8): 312-315.
[13]中国农业科学院畜牧研究所.棉籽饼(粕)营养成分及有毒有害物质含量[M].北京:中国农业科学院畜牧研究所,1990:1-40.
[14]贾晓红, Wedegaertner T.充分利用棉籽粕[J].中国畜牧兽医. 1982, 6: 11-16.
[15]周伟良,尚建波.棉籽饼粕的价值及游离棉酚检测方法[J].江西饲料. 1996, (003): 14-15.
[16]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.
[17]沈维华,马承融.棉籽制油工艺对棉籽蛋白品质的影响[J].中国油脂. 1988, 6: 27-31.
[18] Waldroup P, H. KJ. Nutrient composition of cottonseed meal[J]. Feedstuffs. 2002, 74(45): 11-12.
[19]张贤亮,邹焰峰,孙秋亮.奎屯,石河子垦区部分团场棉籽饼棉酚含量的测定[J].新疆农垦科技. 1990, 4(7).
[20]马春晖,万英.南疆地区部分县,团场棉籽饼棉酚含量的测定[J].塔里木农垦大学学报. 1994, 6(001): 30-32.
[21]李德发.中国饲料大全[M].北京:中国农业出版社, 2001.
[22]王安平.棉粕和棉籽营养成分与棉酚含量测定及其对奶牛产奶量和乳品质的影响[J].湖南农业大学, 2009.
[23] Yang W, Xiang S. Separation of Gossypol Enantiomers byμBondapak C18 Column[M]. Sepu,1995.
[24] Brocas C, Rivera R, Paula-Lopes F, et al. Deleterious actions of gossypol on bovine spermatozoa, oocytes, and embryos[J]. Biology of Reproduction. 1997, 57(4): 901.
[25] Chenoweth P, Chase C. Characterization of gossypol-induced sperm abnormalities in bulls[J]. Theriogenology. 2000, 53(5): 1193-1203.
[26] Yildirim-Aksoy M, Lim C, Wan P, et al. Effect of natural free gossypol and gossypol-acetic acid on growth performance and resistance of channel catfish (Ictalurus punctatus) to Edwardsiella ictaluri challenge[J]. Aquaculture Nutrition. 2004, 10(3): 153-165.
[27] Randel R, Chase Jr C, Wyse S. Effects of gossypol and cottonseed products on reproduction of mammals[J]. Journal of Animal Science. 1992, 70(5): 1628.
[28] Risco CA, Holmberg CA, Kutches A. Effect of Graded Concentrations of Gossypol on Calf Performance: Toxicological and Pathological Considerations1[J]. Journal of dairy science. 1992, 75(10): 2787-2798.
[29] Lordelo M, Davis A, Calhoun M, et al. Relative toxicity of gossypol enantiomers in broilers[J]. Poultry science. 2005, 84(9): 1376.
[30] Blom J, Lee K, Rinchard J, et al. Reproductive efficiency and maternal-offspring transfer of gossypol in rainbow trout (Oncorhynchus mykiss) fed diets containing cottonseed meal[J]. Journal of Animal Science. 2001, 79(6): 1533.
[31] Bender HS, Saunders GK, Misra HP. A histopathologic study of the effects of gossypol on the female rat[J]. Contraception. 1988, 38(5): 585-592.
[32] Bozek SA, Jensen D, Tone J. Scanning electron microscopic study of spermatozoa from gossypol-treated rats[J]. Cell and Tissue Research. 1981, 219(3): 659-663.
[33] Oko R, Hrudka F. Segmental aplasia of the mitochondrial sheath and sequelae induced by gossypol in rat spermatozoa[J]. Biology of Reproduction. 1982, 26(1): 183.
[34] Hoffer AP. Ultrastructural, biochemical and endocrine studies on the effects of gossypol and its isomeric derivatives on the male reproductive tract[J]. Gossypol, a potential contraceptive for men, Plenum Press, New York. 1985: 143?186.
[35] Panigrahi S, Plumb V, Machin D. Effects of dietary cottonseed meal, with and without iron treatment, on laying hens[J]. British poultry science. 1989, 30(3): 641.
[36] Panigrahi S, Hammonds T. Egg discolouration effects of including screw-press cottonseed meal in laying hen diets and their prevention[J]. British poultry science. 1990, 31(1): 107-120.
[37] Ciereszko A, Dabrowski K. In vitro effect of gossypol acetate on yellow perch (Perca flavescens) spermatozoa[J]. Aquatic toxicology. 2000, 49(3): 181-187.
[38] Dabrowski K, Lee KJ, Rinchard J, et al. Gossypol isomers bind specifically to blood plasmaproteins and spermatozoa of rainbow trout fed diets containing cottonseed meal[J]. Biochimica et Biophysica Acta (BBA)-General Subjects. 2001, 1525(1-2): 37-42.
[39] Rinchard J, Lee K, Czesny S, et al. Effect of feeding cottonseed meal-containing diets to broodstock rainbow trout and their impact on the growth of their progenies[J]. Aquaculture. 2003, 227(1-4): 77-87.
[40] Lee KJ, Rinchard J, Dabrowski K, et al. Long-term effects of dietary cottonseed meal on growth and reproductive performance of rainbow trout: Three-year study[J]. Animal feed science and technology. 2006, 126(1-2): 93-106.
[41]张庆朝,王化春.棉酚对畜禽的毒害作用[J].中国兽医科技. 1994, 24(010): 17-18.
[42]卢智文.日粮中棉酚含量计算方法及安全限量[J].中国饲料. 1996, 24: 33-34.
[43] Longmore J. Cottonseed oil: Its coloring matter mucilage, and description of a new method of recovering the loss occurring in the refining process[J]. J. Soc. Chem. Ind. 1886, 5: 200.
[44] Berardi L, Goldblatt C, Liener I. Gossypol[J]. Toxic constituents of plant foodstuffs. 1980: 183-237.
[45] Campbell K, Morris R, Adams R. The Structure of Gossypol. I[J]. Journal of the American Chemical Society. 1937, 59(9): 1723-1728.
[46] Hron R, Koltun S, Pominski J, et al. The potential commercial aspects of gossypol[J]. Journal of the American Oil Chemists' Society. 1987, 64(9): 1315-1319.
[47] Marin A, Shlyapnikov Y, Mametov E, et al. Retardation of polymer oxidation by natural antioxidant gossypol. Part 1: kinetics of oxygen consumption[J]. Polymer degradation and stability. 1992, 35(2): 141-146.
[48]王飞,米海峰.棉籽饼粕中棉酚对动物机体的毒害作用及其预防措施[J].粮食与饲料工业. 2003, (008): 27-28.
[49]白玉萍,于树森,都徐亮.挤压膨化机在棉籽,米糠油料加工中的应用[J].中国油脂. 2004, 29(012): 36-37.
[50] Rhee K, Ziprin Y, Calhoun M. Antioxidative effects of cottonseed meals as evaluated in cooked meat[J]. Meat science. 2001, 58(2): 117-123.
[51] Lusas E, Jividen G. Glandless cottonseed: A review of the first 25 years of processing and utilization research[J]. Journal of the American Oil Chemists' Society. 1987, 64(6): 839-854.
[52]黄祖德.混合溶剂一次浸出棉仁脱酚[J].中国油脂. 1994, 19(002): 25-27.
[53]刘大川,阎杰.溶剂法浸出棉籽油和脱除棉酚的工艺研究[J].中国油脂. 1999, 24(004): 6-8.
[54]戴卫东,李泉,钱礼华,等丙酮,己烷与水混合溶剂脱除棉酚的实验研究[J].化学与生物工程. 2004, 21(005): 32-33.
[55]祝强,刘进才,唐金泉.一种新的棉籽脱酚生产工艺[J].中国油脂. 2004, 29(001): 68-70.
[56] Rahma E, Raoms N. Gossypol removal and functional properties of protein produced byextraction of glanded cottonseed with different solvents[J]. Journal of Food Science. 1984, 49(4): 1057-1060.
[57] Johnson L, Farnsworth J, Sadek N, et al. Pilot plant studies on extracting cottonseed with methylene chloride[J]. Journal of the American Oil Chemists' Society. 1986, 63(5): 647-652.
[58] Abraham G, Decossas K, Hron R, et al. Process engineering economic evaluation of the ethanol extraction of cottonseed: preliminary analysis[J]. Journal of the American Oil Chemists' Society. 1991, 68(6): 418-421.
[59] Lusas EW, Watkins LR, Koseoglu S. Isopropyl alcohol to be tested as solvent[J]. International news on fats, oils and related materials (USA). 1991.
[60] Cherry JP, Gray MS. Methylene chloride extraction of gossypol from cottonseed products[J]. Journal of Food Science. 1981, 46(6): 1726-1733.
[61]乔国平,王立.棉籽饼粕脱毒方法研究进展[J].粮食与油脂. 2002, (003): 27-29.
[62] Kamga R, Kayem GJ, Rouxhet PG. Adsorption of gossypol from cottonseed oil on oxides[J]. Journal of colloid and interface science. 2000, 232(1): 198-206.
[63]石秀侠,程茂基,蔡克周,等.棉籽饼有毒物质及其脱毒方法研究进展[J].饲料博览. 2005, (006): 8-10.
[64]阎轶沽,宋维平,张建云.棉籽饼粕在畜禽中的应用及棉酚的脱毒方法研究[J].饲料工业. 2005, 26(003): 47-50.
[65] Nagalakshmi D, Sastry V, Agrawal D. Detoxification of undecorticated cottonseed meal by various physical and chemical methods[J]. Animal Nutrition and Feed Technology. 2002, 2(2): 117-126.
[66] Nagalakshmi D, Sastry V, Pawde A. Rumen fermentation patterns and nutrient digestion in lambs fed cottonseed meal supplemental diets[J]. Animal feed science and technology. 2003, 103(1-4): 1-14.
[67] Tabatabai F, Golian A, Salarmoeini M. Determination and detoxification methods of cottonseed meal gossypol for broiler chicken rations[J]. Agricultural Sciences and Technology. 2002.
[68]李延云,聂宇燕.棉籽饼粕工业化脱毒生产技术[J].饲料工业. 1999, 20(002): 11-12.
[69]张跃顺.棉籽饼的脱毒方法及其效果[J].山东畜牧兽医. 2004, (001): 13-13.
[70] Barraza M, Coppock C, Brooks K, et al. Iron Sulfate and Feed Pelleting to Detoxify Free Gossypol in Cottonseed Diets for Dairy Cattle1[J]. Journal of dairy science. 1991, 74(10): 3457-3467.
[71]仲明,高庆华.棉籽粕几种脱毒方法效果的比较[J].中国饲料. 1998, (006): 31-32.
[72]张嗣炯.棉籽饼粕脱毒工艺的研究[J].中国粮油学报. 1995, 10(004): 55-58.
[73]高金宝.棉籽料坯高碱水蒸炒[J].中国油脂. 1998, 23(001): 55-56.
[74]戴卫东,卢伯南,钱礼华,等.热碱法脱除游离棉酚的实验研究[J].应用化工. 2004, 33(006): 57-59.
[75]赵兴华,何欣,董强,等.植物毒素的微生物降解与利用研究进展[J].广西农业生物科学. 2006, 25(B09): 172-177.
[76] El-Saidy DMSD, Gaber MM. Use of cottonseed meal supplemented with iron for detoxification of gossypol as a total replacement of fish meal in Nile tilapia, Oreochromis niloticus (L.) diets[J]. Aquaculture Research. 2004, 35(9): 859-865.
[77]杨晓斌,林庆生.微生物饲料添加剂的研究进展[J].氨基酸和生物资源. 2003, 25(002): 10-13.
[78]徐姗楠,邱宏端.微生物发酵生产蛋白饲料的研究进展[J].福州大学学报:自然科学版. 2002, (0z1): 709-713.
[79] Weng XY, Sun JY. Biodegradation of free gossypol by a new strain of Candida tropicalis under solid state fermentation: Effects of fermentation parameters[J]. Process Biochemistry. 2006, 41(7): 1663-1668.
[80] Weng XY, Sun JY. Kinetics of biodegradation of free gossypol by Candida tropicalis in solid-state fermentation[J]. Biochemical Engineering Journal. 2006, 32(3): 226-232.
[81] Reiser R, Fu HC. The mechanism of gossypol detoxification by ruminant animals[J]. The Journal of Nutrition. 1962, 76(2): 215.
[82]钟英长,吴玲娟.棉籽饼的微生物脱毒法研究[J].饲料工业. 1989, (008): 4-8.
[83] Rajarathnam S, Shashirekha M, Bano Z. Biodegradation of gossypol by the white oyster mushroom, Pleurotus florida, during culturing on rice straw growth substrate, supplemented with cottonseed powder[J]. World Journal of Microbiology and Biotechnology. 2001, 17(3): 221-227.
[84]钟英长,吴玲娟.利用微生物将棉籽中游离棉酚脱毒的研究[J].中山大学学报(自然科学版). 1989, 3.
[85]施安辉,黄玉梅.微生物脱毒强化棉籽饼的研究与应用[J].粮食与饲料工业. 1997, (007): 22-23.
[86]孙建义,许梓荣.利用假丝酵母进行棉籽饼脱毒的研究[J].中国粮油学报. 1995, 10(001): 61-64.
[87]顾赛红,孙建义,李卫芬.黑曲霉PES固体发酵对棉籽粕营养价值的影响[J]. JOURNAL OF THE CHINESE CEREALS AND OILS ASSOCIATION. 2003, 18(1).
[88]张文举,许梓荣,孙建义,等.假丝酵母ZD-3与黑曲霉ZD-8复合固体发酵对棉籽饼脱毒及营养价值的影响研究[J].中国粮油学报. 2006, 21(006): 129-135.
[89]院江,孙新文,丁宁,等.微生物发酵对棉籽壳营养成分及游离棉酚的影响[J].石河子大学学报. 2006, 24(003): 299-301.
[90]张庆华,赵新海,钟丽娟,等.三菌株协同固态发酵对棉粕脱毒效果及其生物活性的影响[J].饲料工业. 2007, 28(018): 37-38.
[91] Yude H. Effect of degossypolized cottonseed meal by microbial method on the performance of layers[J]. Chinese Journal of Animal Science (China). 1994.
[92] Marquie C, Aymard C, Cuq JL, et al. Biodegradable packaging made from cottonseed flour:formation and improvement by chemical treatments with gossypol, formaldehyde, and glutaraldehyde[J]. Journal of Agricultural and Food Chemistry. 1995, 43(10): 2762-2767.
[93] Shashirekha M, Rajarathnam S, Bano Z. Enhancement of bioconversion efficiency and chemistry of the mushroom, Pleurotus sajor-caju (Berk and Br.) Sacc. produced on spent rice straw substrate, supplemented with oil seed cakes[J]. Food chemistry. 2002, 76(1): 27-31.
[94] Zhang WJ, Xu ZR, Zhao SH, et al. Optimization of process parameters for reduction of gossypol levels in cottonseed meal by Candida tropicalis ZD-3 during solid substrate fermentation[J]. Toxicon. 2006, 48(2): 221-226.
[95] Zhang WJ, Xu ZR, Zhao SH, et al. Development of a microbial fermentation process for detoxification of gossypol in cottonseed meal[J]. Animal feed science and technology. 2007, 135(1-2): 176-186.
[96]吴小月,陈金湘.利用微生物降解棉仁饼粕中游离棉酚的研究[J].中国农业科学. 1989, 22(2): 82-86.
[97]杨景芝,孙衍华.棉酚脱毒微生物的筛选及其脱毒效果的研究[J].山东农业大学学报:自然科学版. 1999, 30(001): 26-30.
[98]何涛,滕达,张海军,等.微生物降解棉酚的响应面分析优化[J].食品与发酵工业. 2008, 34(004): 71-74.
[99]吴伟伟,许赣荣.复合微生物固态发酵对棉籽饼粕脱毒及营养的影响[J]. INDUSTRIAL MICROBIOLOGY. 2010, 40(3).
[100] Smith F. Determination of gossypol in leaves and flower buds of Gossypium[J]. Journal of the American Oil Chemists' Society. 1967, 44(4): 267-269.
[101]宁开桂.实用饲料分析手册[M]..中国农业科技出版社, 1993.
[102] Lee KJ, Dabrowski K. High-performance liquid chromatographic determination of gossypol and gossypolone enantiomers in fish tissues using simultaneous electrochemical and ultraviolet detectors[J]. Journal of Chromatography B. 2002, 779(2): 313-319.
[103]李信,刘云,计林贞.侧孢霉利用玉米秸秆固体发酵产生木质纤维素酶的研究[J].核农学报. 2000, 14(2): 99-103.
[104]张文举.高效降解棉酚菌种的选育及棉籽饼粕生物发酵的研究[J].浙江大学, 2006.
[105]院江,孙新文,蔡铁查.棉籽壳发酵前后游离棉酚及营养成分变化的实验[J].饲料研究. 2006, (004): 3-4.
[106]孙中涛.苹果渣与棉粕固态发酵生产NSP酶的研究[D].山东农业大学, 2008.
[107]布坎南RE.伯杰氏细菌鉴定手册[M].北京:科学出版社, 1988.
[108]贾晓锋.固态发酵对棉籽粕棉酚脱毒及蛋白质降解的影响[J].西北农林科技大学, 2008.
[109] Jia Xiaofeng NA. Effect of solid-state fermentation on gossypol detoxification and protein degradation in cottonseed meal[J]. Journal of Northwest Agriculture and Forestry University. 2009, 37.
[110]汤江武,孙宏,姚晓红,等.响应面法与基于神经网络的遗传算法在优化棉粕固态发酵中外源添加物用量上的应用[J].浙江大学学报(农业与生命科学版). 2011, 37(1): 97-102.
[111] Murray BE, Singh KV, Markowitz SM, et al. Evidence for clonal spread of a single strain ofβ-lactamase-producing Enterococcus (Streptococcus) faecalis to six hospitals in five states[J]. Journal of Infectious Diseases. 1991, 163(4): 780.
[112]刘凤强,姬生瑞.检出160株肠球菌的分布及耐药性分析[J].中国热带医学. 2005, 5(009): 1879-1879.
[113] Murray BE. The life and times of the Enterococcus[J]. Clinical Microbiology Reviews. 1990, 3(1): 46.
[114]段永兰.益生菌与动物饲料[J].郑州牧业工程高等专科学校学报. 2007, 27(002): 31-33.
[115] Dimov SG. A novel bacteriocin-like substance produced by Enterococcus faecium 3587[J]. Current Microbiology. 2007, 55(4): 323-327.
[116] Strompfova V, Laukova A, Ouwehand AC. Selection of enterococci for potential canine probiotic additives[J]. Veterinary microbiology. 2004, 100(1-2): 107-114.
[117] Settanni L, Valmorri S, Suzzi G, et al. The role of environmental factors and medium composition on bacteriocin-like inhibitory substances (BLIS) production by Enterococcus mundtii strains[J]. Food microbiology. 2008, 25(5): 722-728.
[118] Callewaert R, Hugas M, Vuyst LD. Competitiveness and bacteriocin production of Enterococci in the production of Spanish-style dry fermented sausages[J]. International Journal of Food Microbiology. 2000, 57(1-2): 33-42.
[119] Moon GS, Kang CH, Pyun YR, et al. Isolation, identification, and characterization of a bacteriocin-producing Enterococcus sp. from kimchi and its application to kimchi fermentation[J]. Journal of microbiology and biotechnology. 2004, 14(5): 924-931.
[120] Kim J, Oh M, Rhee Y, et al. Selection and physico-chemical characteristics of lactic acid bacteria which had cholesterol lowering activities[J]. The Korean Society of Agricultural Chemistry and Biotechnology. 1999.
[121] Hlivak P, Odraska J, Ferencik M, et al. One-year application of probiotic strain Enterococcus faecium M-74 decreases serum cholesterol levels[J]. Bratisl Lek Listy. 2005, 106(2): 67-72.
[122] Martínez?Bueno M, Valdivia E, Gálvez A, et al. Analysis of the gene cluster involved in production and immunity of the peptide antibiotic AS?48 in Enterococcus faecalis[J]. Molecular microbiology. 1998, 27(2): 347-358.
[123] Benyacoub J, Czarnecki-Maulden GL, Cavadini C, et al. Supplementation of food with Enterococcus faecium (SF68) stimulates immune functions in young dogs[J]. The Journal of Nutrition. 2003, 133(4): 1158.
[124] Hollis A, Wilkins P, Palmer J, et al. Bacteremia in equine neonatal diarrhea: a retrospectivestudy [J]. Journal of Veterinary Internal Medicine. 2008, 22(5): 1203-1209.
[125] Knorr R, Cavadini C, Benyacoub J, et al. Probiotic Enterococci For Improved Immunity [M]. EP Patent, 2010.
[126] Tanaka NG, Sakamoto N, Inoue K , et al. Antitumor effects of an antiangiogenic polysaccharide from an Arthrobacter species with or without a steroid[J]. Cancer research. 1989, 49(23): 6727.
[127] Xiong G, Maser E. Regulation of the steroid-inducible 3α-hydroxysteroid dehydrogenase /carbonyl reductase gene in Comamonas testosteroni[J]. Journal of Biological Chemistry. 2001, 276(13): 9961.
[128] Metchnikoff E, Metchnikoff II, Mitchell SPC. The prolongation of life: optimistic studies[m]. Springer Publishing Company, 2004.
[129]孔祥武,钮跃贞,陈圣洁,等.微生态制剂的临床应用[J].中华实用医药杂志. 2003, 3(6).
[130] Fioramonti J, Theodorou V, Bueno L. Probiotics: what are they? What are their effects on gut physiology?[J]. Best Practice & Research Clinical Gastroenterology. 2003, 17(5): 711-724.
[131] Alonso L, Cuesta E, Gilliland S. Production of Free Conjugated Linoleic Acid by Lactobacillus acidophilus and Lactobacillus casei of Human Intestinal Origin1[J]. Journal of dairy science. 2003, 86(6): 1941-1946.
[132] Furushiro M, Sawada H, Hirai K, et al. Blood Pressure-lowering Effect of Extract from Lactobacillus casei in Spontaneously Hypertensive Rats (SHR)(Microbiology & Fermentation Industry)[J]. Agricultural and biological chemistry. 1990, 54(9): 2193-2198.
[133] Liong M, Shah N. Acid and bile tolerance and cholesterol removal ability of lactobacilli strains[J]. Journal of dairy science. 2005, 88(1): 55-66.
[134] Aso Y, Akaza H, Kotake T , et al. Preventive effect of a Lactobacillus casei preparation on the recurrence of superficial bladder cancer in a double-blind trial[J]. European urology. 1995, 27(2): 104-109.
[135] Reid G. The scientific basis for probiotic strains of Lactobacillus[J]. Applied and environmental Microbiology. 1999, 65(9): 3763.
[136] Zhang R, Xu Y, Sun Y , et al. Construction of an enzyme-coupled system consisting of (R)-and (S)-specific carbonyl reductases for one-step preparation of (S)-1-phenyl-1, 2-ethanediol[J]. Wei sheng wu xue bao= Acta microbiologica Sinica. 2009, 49(2): 204.
[137]褚以文.微生物培养基优化方法及其OPTI优化软件[J].国外医药:抗生素分册. 1999, 20(002): 58-60.
[138] Ahuja S, Ferreira G, Moreira A. Application of Plackett?Burman design and response surface methodology to achieve exponential growth for aggregated shipworm bacterium[J]. Biotechnology and bioengineering. 2004, 85(6): 666-675.
[139] Loukas YL. A Plackett-Burnam screening design directs the efficient formulation ofmulticomponent DRV liposomes[J]. Journal of pharmaceutical and biomedical analysis. 2001, 26(2): 255-263.
[140] Hanrahan G, Lu K. Application of factorial and response surface methodology in modern experimental design and optimization[J]. Critical Reviews in Analytical Chemistry. 2006, 36(3-4): 141-151.
[141]杨承剑,黄兴国,李伟,等. Plackett-Burman设计在益生菌生长主要影响因子筛选中的应用[J].饲料工业. 2007, 28(016): 31-33.
[142] Vindevogel J, Sandra P. Resolution optimization in micellar electrokinetic chromatography: use of Plackett-Burman statistical design for the analysis of testosterone esters[J]. Analytical Chemistry. 1991, 63(15): 1530-1536.
[143] Ghanem NB, Yusef HH, Mahrouse HK. Production of Aspergillus terreus xylanase in solid-state cultures: application of the Plackett-Burman experimental design to evaluate nutritional requirements[J]. Bioresource technology. 2000, 73(2): 113-121.
[144] Rao P, Divakar S. Lipase catalyzed esterification of [alpha]-terpineol with various organic acids: application of the Plackett-Burman design[J]. Process Biochemistry. 2001, 36(11): 1125-1128.
[145] Khandeparkar R, Bhosle N. Isolation, purification and characterization of the xylanase produced by Arthrobacter sp. MTCC 5214 when grown in solid-state fermentation[J]. Enzyme and microbial technology. 2006, 39(4): 732-742.
[146] Walker GM. Metals in yeast fermentation processes[J]. Advances in applied microbiology. 2004, 54: 197-229.
[147] Puri M, Banerjee A, Banerjee U. Optimization of process parameters for the production of naringinase by Aspergillus niger MTCC 1344[J]. Process Biochemistry. 2005, 40(1): 195-201.
[148] Prawirodigdo S, Gannon N, Van Barneveld R , et al. Assessment of apparent ileal digestibility of amino acids and nitrogen in cottonseed and soyabean meals fed to pigs determined using ileal dissection under halothane anaesthesia or following carbon dioxide-stunning[J]. British Journal of Nutrition. 1998, 80(02): 183-191.
[149] Knabe D, LaRue D, Gregg E , et al. Apparent digestibility of nitrogen and amino acids in protein feedstuffs by growing pigs[J]. Journal of Animal Science. 1989, 67(2): 441.
[150]杨继良,周大云,杨伟华,等.高效降解棉酚菌种的筛选及棉饼脱毒参数的研究[J].棉花学报. 2002, 12(5): 225-229.
[151]肖永友,姜玉,王沦娟,蔡发国.高效降解棉酚菌株的筛选及其发酵技术研究[J].广东饲料. 2008, 3:34-36.
[152]朱德伟,刘志鹏,陆建,等.高效降解棉酚菌种的筛选及棉粕发酵脱毒工艺研究[J].中国油脂. 2010 ,35 (2): 24-28.
[153]聂蓬勃,汤江武,梁运祥.棉粕脱毒菌株的筛选及发酵条件的研究[J].浙江农业科学. 2009, 1:120-122.
[154] Pettit R K. Mixed fermentation for natural product drug discovery[J]. Applied microbiology and biotechnology. 2009, 83(1): 19-25.
[155] Schneider IC, 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: 2267-2273.
[156] Abou-Donia MB, Lyman CM, Dieckert JW. Metabolic fate of gossypol: the metabolism of 14C-gossypol in rats[J]. Lipids. 1970,5: 938-946.
[157] Abou-Donia MB, Dieckert JW. Metabolic fate of gossypol: the metabolism of [14C]gossypol in swine[J]. Toxicology and Applied Pharmacology. 1975,31: 32-46.
[158] Bajpai M, Gupta G, Setty BS. Changes in carbohydrate metabolism of testicular germ cells during meiosis in the rat[J]. European Journal of Endocrinology. 1998, 138: 322-327.