酸马奶酒中抗菌物质的分离及抗菌作用的研究
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摘要
研究了内蒙古自治区的传统民族乳制品——酸马奶酒的抗菌性，并对抗菌因子进行了分离。参照抗生素效价生物测定法，用牛津杯法测定了其抗菌性。探讨了影响抗菌作用检测效果的因素，研究表明：指示菌及其浓度、琼脂量、培养时间、培养基厚度等因素均对抑菌结果有影响。选择最佳的实验条件，从而提高了抑菌实验的灵敏度与准确性。测定了酸马奶酒中有机酸和酒精的组成，据此配制了人工模拟酸马奶酒，评价了酸马奶酒中有机酸、酒精对细菌的作用。结果表明：有机酸对枯草芽胞杆菌有抑菌作用，其中乳酸起主要作用，醋酸和酒精起增效作用。在抑菌作用中实际起作用的是有机酸的未解离部分。当乳酸和醋酸其未解离部分的浓度分别大于0.683％和0.256％时对枯草芽胞杆菌有抑菌作用；酸马奶酒中含有多种抗菌物质，将酸马奶酒脂肪分为中性与极性区分物，其中中性区分物可明显抑制结核菌。经气相色谱仪测定，中性和极性脂肪的脂肪酸组成上没有明显的差别。因此，不能确定是何种组分在起作用；经超滤分离后，酸马奶酒乳清中分子量在10k以上的组分对枯草芽胞杆菌具有明显的抑制作用，分子量在50k以上的组分最为明显；经层析分离后，分离出一种抗菌组分，电泳检测呈一条带，分子量大约在15，400。其特性还有待于进一步研究。
koumiss, a traditional and national dairy product in the Inner Mongolia, was researched.The antibiotic factors were preliminarily isolated from koumiss. Its antibiotic activity was detected by cylinder-plate method,which is similar to the determination of antibiotic liter. In order to get the best inhibitory effect on bacteria, Some factors that affected determination were analyzed. Many factors including density of the target strains, concentration of agar, culture time, thickness of the medium have effect to computation.The sensitivity and accuracy of the inhibitory test were improved. This study have also evaluated the inhibitory power of koumiss and analyzed the role of organic acids in bacterial inhibition. The results showed that organic acids in koumiss had apparent inhibitory effect to bacillus subtilis.Lactic acid was important for inhibition, the presence of acetic acid and alcohol improved the inhibitory effect. This inhibitory effect could be assigned to the undissociated form of organic acids. L
actic acid and acetic acid showed an inhibitory effect toward bacillus subtilis if the undissociated form was more than 0.683% and 0.256%, respectively. There are more than one antibacterial substances in the koumiss. If the fat of koumiss was separated into the neutral fat and the polary, we found that the neutral fat had a very effective inhibition to tubercle bacillus. By chromatography analyzing, there was no significant difference in fatty acid composition between the neutral fat and the polary. After ultra-filtrated, some substances in the whey of koumiss, which molecular weight was more than 10KD, had a powerful inhibitory effect to bacillus subtilis. By chromatography, an antibacterial substance was obtained, which molecular weight was about 15,400 . A further research were still needed.
koumiss, a traditional and national dairy product in the Inner Mongolia, was researched.The antibiotic factors were preliminarily isolated from koumiss. Its antibiotic activity was detected by cylinder-plate method,which is similar to the determination of antibiotic liter. In order to get the best inhibitory effect on bacteria, Some factors that affected determination were analyzed. Many factors including density of the target strains, concentration of agar, culture time, thickness of the medium have effect to computation.The sensitivity and accuracy of the inhibitory test were improved. This study have also evaluated the inhibitory power of koumiss and analyzed the role of organic acids in bacterial inhibition. The results showed that organic acids in koumiss had apparent inhibitory effect to bacillus subtilis.Lactic acid was important for inhibition, the presence of acetic acid and alcohol improved the inhibitory effect. This inhibitory effect could be assigned to the undissociated form of organic acids. L
actic acid and acetic acid showed an inhibitory effect toward bacillus subtilis if the undissociated form was more than 0.683% and 0.256%, respectively. There are more than one antibacterial substances in the koumiss. If the fat of koumiss was separated into the neutral fat and the polary, we found that the neutral fat had a very effective inhibition to tubercle bacillus. By chromatography analyzing, there was no significant difference in fatty acid composition between the neutral fat and the polary. After ultra-filtrated, some substances in the whey of koumiss, which molecular weight was more than 10KD, had a powerful inhibitory effect to bacillus subtilis. By chromatography, an antibacterial substance was obtained, which molecular weight was about 15,400 . A further research were still needed.
引文
1．乌尼，敖敦格日勒等．内蒙古牧区民族乳制品的种类及制造工艺．内蒙古农牧学院学报，1996，17(1)：63—68
2．秦立虎．我国少数民族的马奶酒初探．中国酿造，1998，(3)：1—6
3．杨晓春．蒙、元时期的马奶酒考．西北民族研究．1999，(1)：15
4．郭福德．肃北玉浆-马奶酒，中国土特产．1995，(3)：21
5．郭本恒著．乳品微生物学．北京：中国轻工业出版社，2001，7．285
6．杨茉莉．马奶成分分析及开发利用现状．陕西农业科学，2001，(1)：13—15
7．汪长明，吕军．初露端倪的马奶加工业．新疆畜牧业，1999，2：23～24
8．候文通．马奶脂肪抗结核杆菌试验．畜牧兽医杂志，1989，(2)：1—4
9．候文通等．关中马奶营养成分分析．西北农业大学学报，1988，16(3)75—79
10．郭本恒著．乳品化学．北京：中国轻工业出版社，2001，7．329—331
11．嘎日迪．马奶酒中含的有益人体的金属元素．内蒙古石油化工．1995，2：6—7
12．雷天富等．马奶啤酒的试验研究．中国乳品工业，1995，23(3)：108—110
13．金花，代利君等．酸马奶．中国乳品工业．1998，26(4)：16—17
14．呼春，达古拉．蒙古族的酸马奶酒疗法．中国民族医药杂志，1996，2(1)：32-33
15．关加怀．马奶及酸马奶酒的医疗作用．中国畜产与食品，1998，(2)：87-88
16．扎木苏，刘月英等．酸马奶降脂及抗凝作用50例临床观察．内蒙古中医药，1995，1：16
17．刘忠贤，马丽亚汉．马奶酒治疗便秘．中国民间疗法．2000，8(6)：27—28
18．查哈达．酸马奶治疗肺结核56例临床观察．中国民族医药杂志，1999，36(4)，13—14
19．王泽之．酸马奶的食疗价值．当代畜禽养殖，1999，(7)：28
20．包德毕力格等．策格(酸马奶)霜倒模面模治疗黄褐斑44例疗效观察．中国皮肤性病学杂志．1998，2
21．骆承庠主编．乳与乳制品工艺学．第二版．北京：中国农业出版社，2001，2．158
22．谢继志主编．液态乳制品科学与技术．北京：中国轻工业出版社，1999，8．244
23．张列兵，吕加平．新版乳制品配方。北京：中国轻工业出版社，2002，1．118
24．郭本恒著．功能性乳制品．北京：中国轻工业出版社，2001，2．109—121
25．张和平等．利用开菲尔发酵剂制备牛乳酸马奶酒的研究．中国乳品工业．1996，24(4)：16—20
26．贺银凤，李少英．酸马奶酒中微生物的分离鉴定及抗菌特性的研究．农业工程学报，2002，18(2)91—95
27．F．M．德赖森，Z．普汉．发酵乳科学与技术．顾瑞霞等译．广州：东南大学出版社，1991
28．顾瑞霞．乳与乳制品的生理功能特性．中国轻工业出版社，2000，1．311
29．汪玉松，邹思湘．乳生物化学．吉林大学出版社．1995，12
30．hson Wan，Karl Mccann．牛奶中的抗菌及抗病毒生物活性物质．中国乳制品工业协会第七次年会论文集，1999，157—164
31．张和平。免疫乳及其制品的研究。中国乳品工业，1997，25(6)：15-25
32．金世琳。古老而新型的酒精性发酵乳饮料—开菲尔．中国乳品工业，1999，27(2)：18—23
33．姜成林，徐丽华．微生物资源的开发利用．中国轻工业出版社，2001，6
34．张列兵．Kefir的营养特性及保键作用．中国乳品工业，1987，25(2)：36—38
35．李平兰．产细菌素植物乳杆菌菌株的筛选及细菌素生物学特性研究．食品与发酵工业，1999，25(1)：1—4
36．温旺荣，陈红等．酵母菌杀菌系统的杀菌活性．中国微生态学杂志，1999，11(4)：217—219
37．秦玉静，高东．嗜杀酿酒酵母毒素蛋白及其杀伤质粒的研究，微生物学报，2000，40(1)：1051-107
38．秦玉静，刘巍峰等．酿酒酵母两种不同嗜杀菌株的比较研究．微生物学报，1998，38(2)：92～97
39．巴图等．巯基羊毛吸附分离-分光光度法测定马奶酒中的微量锌．内蒙古师范大学学报(自然科学汉文版)．1997，3
40．中华人民共和国药典．一九八五年版二部，人民卫生出版社，1985，2．附录95-98
41．陈天寿．培养基的制作与应用．北京：中国农业出版社，1995，6．269—293
42．刘稳，朱文森等．一株产乳酸链球菌素菌株SM526的分离鉴定及生理特性研究．山东大学学报(自然科学版)，1997，32(1)：113—121
43．贵州农学院主编．生物统计附试验设计．第二版．北京：中国农业出版社 1999，7．262—268
44．李建武等编．生物化学实验原理和方法．北京：北京大学出版社，1994，9．36—56
45．张龙翔编．生物化学实验方法和技术．北京：人民教育出版社，1981．206
46．沈同，王镜岩主编．生物化学．北京：高等教育出版社，1991．209—211，347—348
47．郭尧君著．蛋白质电泳实验技术．北京：科学出版社，1999，9．123—157
48．何忠效，张树政主编．电泳．第二版．北京：科学出版社，1999，2．127—139
49．A．Lomascolo．对乳制品中脂肪酸组成的观察结果．赵敏．中国乳品工业，1995，23(6)：297-301
50．杨桂梧主编。普通化学，沈阳：辽宁民族出版社，1996，85
51. Gladkova,E.sources of health drinks. Feeding of mares and preparation of koumiss on farms. Konevodstvo I konnyisport, 1992,No. 1:4-5
52. Koroleva,N.S.Technology of kefir and kumiss. Bull.inter. dairy fed. 1988,227:96-100
53. Chandan,R.C.;Shahani,K.M.Other fermented dairy products. Biotechnology. Volume 9:enzymes, biomass, food and feed., 1995:385-418
54. Tamime, A. Y.;Muir, D. D.; Wasolek,M. kefir, koumiss and kishk. Dairy Industries international, 1999, Vol64,No.5:32-33
55. Habe,F. Yield and possibilities for the use of mares' milk. Stocarstvo,2000,Vol.54:295-304
56. Doreau,M. mare milk. Equ'idee-Bulletin d'information sur les equities, 1992,No.6:55-63
57. Doreau,M. mare milk.productions animals, 1991,Vol.4,No.4:297-302
58. Emanuele M.,Gianfranco p. chemical composition and nutritional properties of commercial products of mare powder. Journal of food composition and analysis. 1998,11:178-187
59. Valieva,T.A.;Valiev,A.G. Lipid and fatty acid contents, and lipid peroxidation in freeze-dries mare milk during prolonged storage with antioxidants. Voprosy Pitaniya, 1993,No.5:61-65
60. Valiev, A.G.;Valieva,T.A.etc.the influence of essential fatty acids of mare's milk on the immune system and nonspecific resistance of rats. Voprosy Pitaniya, 1999,Vol.68,No.3:3-6
61. Prateek jain; Gupta, S.K.nutritional potentiality of mares'milk. Indian dairyman, 1998,Vol.50,No.8:39-42
62. Fantova,M.; kukanova,J. Possibilities of utilization of mares' milk. Agriculture tropica et subtropica, 1998,No.31:177-181
63. FrancescoB.StefaniaL.E.Thermal sensitivity of mares'milk proteins. Journal of dairy research. 1994,61:419-422
64. Lozovich,S. Medical uses of whole and fermented mare milk in Russia. Cultured Dairy Products Journal, 1995,Vol.30,No. 1:18-19
65. Szakaly, S.Role of fermented milk products in protection of human health. Tejgazdasag, 1999, Vol59,No.2:15-18
66. Y. Tamime. special feature-kefir, koumiss and kishk. Dairy industries international. 1999, Vol. 64,No. 5:32-33
67. Mistry, V.V. Fermented milks and cream. Applied dairy microliology, 2001,No.Ed.2:301-325
68. Prehrambene, J.L.Samarzija,D. Nutritional, health and technological properties of cultured products.Mljekarstvo, 1996,Vol.46,No.4:265-273
69. Storch G, Composition and properties of mares' milk and koumiss,with particular regard to dietetic aspects. Thesis,Justus-Liebig-Universitat Giessen,German,Federal Republic 1985,98
70. Omel'chenko,N.P. Nutrition of breast-feeding women in Yakut conditions. Voprosy Pitaniya, 1993,No. 1:23-27
71. Montanari,G.Zambonelli,C.Koumiss,a cultured milk drink.lndustrie Alimentari, 1997,Vol.36,No.355:5-9
72. Molochnikov, V.V.;Anisimov, S.V. Mixture for koumiss and manufacturing method. USSR,Patent, 1991,No. SU 1 664 238
73. Pastukhova Z M, Gerbeda V V. Conparative composition of lipids of mares'milk and of a koumiss mixture based on cows' milk. FSTA, 1982
74. Robinson, R.K.; Tamime,A.Y. Microbiology of fermented milks. Dairy microbiology handbook:the microbiology of milk and milk products,2002,No.Ed:367-430
75. Ishii,S. Survey of the koumiss of Sakha. Milk Science, 1999, Vol48,No.2:193-195
76. Gregurek,L.Dairy cultures of microorganisms in the manufacture of cultured milks. Mljekarstvo, 1997,Vol.47,No.2:103-113
77. Ishii, S.; Kikuchi, M.; Takao, S. Isolation and identification of lactic acid bacteria and yeasts from "Chigo" in Inner Mongolia, China. Animal Science and Technology, 1997,Vol. 68,No. 3:325-329
78. Montanari ,G.; Zambonelli, C.; Grazia, L. Saccharomyces unisporus as the principal alcohol fermentation microorganism of traditional koumiss, etal. J. of Dairy Research, 1996, Vol.63,No. 2:327-331
79. Ham, J. S.; Jeong, J. Y. etal. Effects of feeding with Lactobacillus plantarum and Candida Kefyr isolated from Mongoiian koumiss on the growth and fecal microflora of broilers. Korean Journal of Dairy Science ,1999,Vol. 21,No. 3:241-246
80. Ham,J.S.etl.fermentation of goat milk using Lactobacillus plantarurn and Candida Kefyr isolated from Mongolian koumiss. Korean Journal of Dairy Science, 1999,Vol. 21,No. 3:247-254
81. Ishii,S. Kikuchi, M.; Takao, S.Identification of compounds causing symbiotic growth of Lactobacillus paracasei subsp. Tolerans and kluyveromyces marxianus var. lactis in chigo, inner Mongolia, china. Animal science Journal, 1999,Vol.70,No.2:81-89
82. Hiroshi shimizu,taiji mizuguchi. Nisin production by mixed-culture systerm consisting of Lactococcus lactis and Kluyveromyces marxianus. Applied and environmental microbiology, 1999,4:3134-3141
83. Jain, A.k.; Prajapati,j.B. Associative growth in dairy starter cultures, jounal of dairying,foods and home sciences, 1998,Vol. 17,No. 1:1-11
84. Lukerchenko,V.N. A new strain of yeast for lactose fermentation. Pishchevaya Promyshlennost, 2001,No.2:58
85. Libudzisz, Z.intestinal microflora and new generations of fermented milk beverages.
Zywienie Czlowieka I Metabolizm, 1999,Vol.26,No. Suppl:5-13
86. Gladkova,E. How to organize koumiss production on the farm. Konevodstvo I Konnyisport, 1997,No.4:8-9
87. Marshall,V.M. starter cultures for milk fermentation and their characteristics. Journal of the society of dairy technology, 1993,Vol.46,No.2:49-56
88. Fleet, G.H.Yeasts in dairy products.The Journal of Applied Bacteriology, 1990,68:199-121
89. Graciela L. Garrote,Analia G. Inhibitory power of kefir: the role of organic acid. J. of food protection. 2000, Vol 63,No.3:364-369
90. Helander, I.M.,A.Von Wright, and T.M. Potential of lactic acid bacteria and novel antimicrobials against gram-negative bacteria Mattila-Sandholm. Trends Food Sci. Technol. 1997,8:146-150
91. Presser. K.A.,D.A. Ratkowsky. and T. Ross. Modlling the growth rate of Escherichia coli as a function of pH and lactic acid concentration. Appl. Environ. Microbiol, 1997,63: 2355-2360
2．秦立虎．我国少数民族的马奶酒初探．中国酿造，1998，(3)：1—6
3．杨晓春．蒙、元时期的马奶酒考．西北民族研究．1999，(1)：15
4．郭福德．肃北玉浆-马奶酒，中国土特产．1995，(3)：21
5．郭本恒著．乳品微生物学．北京：中国轻工业出版社，2001，7．285
6．杨茉莉．马奶成分分析及开发利用现状．陕西农业科学，2001，(1)：13—15
7．汪长明，吕军．初露端倪的马奶加工业．新疆畜牧业，1999，2：23～24
8．候文通．马奶脂肪抗结核杆菌试验．畜牧兽医杂志，1989，(2)：1—4
9．候文通等．关中马奶营养成分分析．西北农业大学学报，1988，16(3)75—79
10．郭本恒著．乳品化学．北京：中国轻工业出版社，2001，7．329—331
11．嘎日迪．马奶酒中含的有益人体的金属元素．内蒙古石油化工．1995，2：6—7
12．雷天富等．马奶啤酒的试验研究．中国乳品工业，1995，23(3)：108—110
13．金花，代利君等．酸马奶．中国乳品工业．1998，26(4)：16—17
14．呼春，达古拉．蒙古族的酸马奶酒疗法．中国民族医药杂志，1996，2(1)：32-33
15．关加怀．马奶及酸马奶酒的医疗作用．中国畜产与食品，1998，(2)：87-88
16．扎木苏，刘月英等．酸马奶降脂及抗凝作用50例临床观察．内蒙古中医药，1995，1：16
17．刘忠贤，马丽亚汉．马奶酒治疗便秘．中国民间疗法．2000，8(6)：27—28
18．查哈达．酸马奶治疗肺结核56例临床观察．中国民族医药杂志，1999，36(4)，13—14
19．王泽之．酸马奶的食疗价值．当代畜禽养殖，1999，(7)：28
20．包德毕力格等．策格(酸马奶)霜倒模面模治疗黄褐斑44例疗效观察．中国皮肤性病学杂志．1998，2
21．骆承庠主编．乳与乳制品工艺学．第二版．北京：中国农业出版社，2001，2．158
22．谢继志主编．液态乳制品科学与技术．北京：中国轻工业出版社，1999，8．244
23．张列兵，吕加平．新版乳制品配方。北京：中国轻工业出版社，2002，1．118
24．郭本恒著．功能性乳制品．北京：中国轻工业出版社，2001，2．109—121
25．张和平等．利用开菲尔发酵剂制备牛乳酸马奶酒的研究．中国乳品工业．1996，24(4)：16—20
26．贺银凤，李少英．酸马奶酒中微生物的分离鉴定及抗菌特性的研究．农业工程学报，2002，18(2)91—95
27．F．M．德赖森，Z．普汉．发酵乳科学与技术．顾瑞霞等译．广州：东南大学出版社，1991
28．顾瑞霞．乳与乳制品的生理功能特性．中国轻工业出版社，2000，1．311
29．汪玉松，邹思湘．乳生物化学．吉林大学出版社．1995，12
30．hson Wan，Karl Mccann．牛奶中的抗菌及抗病毒生物活性物质．中国乳制品工业协会第七次年会论文集，1999，157—164
31．张和平。免疫乳及其制品的研究。中国乳品工业，1997，25(6)：15-25
32．金世琳。古老而新型的酒精性发酵乳饮料—开菲尔．中国乳品工业，1999，27(2)：18—23
33．姜成林，徐丽华．微生物资源的开发利用．中国轻工业出版社，2001，6
34．张列兵．Kefir的营养特性及保键作用．中国乳品工业，1987，25(2)：36—38
35．李平兰．产细菌素植物乳杆菌菌株的筛选及细菌素生物学特性研究．食品与发酵工业，1999，25(1)：1—4
36．温旺荣，陈红等．酵母菌杀菌系统的杀菌活性．中国微生态学杂志，1999，11(4)：217—219
37．秦玉静，高东．嗜杀酿酒酵母毒素蛋白及其杀伤质粒的研究，微生物学报，2000，40(1)：1051-107
38．秦玉静，刘巍峰等．酿酒酵母两种不同嗜杀菌株的比较研究．微生物学报，1998，38(2)：92～97
39．巴图等．巯基羊毛吸附分离-分光光度法测定马奶酒中的微量锌．内蒙古师范大学学报(自然科学汉文版)．1997，3
40．中华人民共和国药典．一九八五年版二部，人民卫生出版社，1985，2．附录95-98
41．陈天寿．培养基的制作与应用．北京：中国农业出版社，1995，6．269—293
42．刘稳，朱文森等．一株产乳酸链球菌素菌株SM526的分离鉴定及生理特性研究．山东大学学报(自然科学版)，1997，32(1)：113—121
43．贵州农学院主编．生物统计附试验设计．第二版．北京：中国农业出版社 1999，7．262—268
44．李建武等编．生物化学实验原理和方法．北京：北京大学出版社，1994，9．36—56
45．张龙翔编．生物化学实验方法和技术．北京：人民教育出版社，1981．206
46．沈同，王镜岩主编．生物化学．北京：高等教育出版社，1991．209—211，347—348
47．郭尧君著．蛋白质电泳实验技术．北京：科学出版社，1999，9．123—157
48．何忠效，张树政主编．电泳．第二版．北京：科学出版社，1999，2．127—139
49．A．Lomascolo．对乳制品中脂肪酸组成的观察结果．赵敏．中国乳品工业，1995，23(6)：297-301
50．杨桂梧主编。普通化学，沈阳：辽宁民族出版社，1996，85
51. Gladkova,E.sources of health drinks. Feeding of mares and preparation of koumiss on farms. Konevodstvo I konnyisport, 1992,No. 1:4-5
52. Koroleva,N.S.Technology of kefir and kumiss. Bull.inter. dairy fed. 1988,227:96-100
53. Chandan,R.C.;Shahani,K.M.Other fermented dairy products. Biotechnology. Volume 9:enzymes, biomass, food and feed., 1995:385-418
54. Tamime, A. Y.;Muir, D. D.; Wasolek,M. kefir, koumiss and kishk. Dairy Industries international, 1999, Vol64,No.5:32-33
55. Habe,F. Yield and possibilities for the use of mares' milk. Stocarstvo,2000,Vol.54:295-304
56. Doreau,M. mare milk. Equ'idee-Bulletin d'information sur les equities, 1992,No.6:55-63
57. Doreau,M. mare milk.productions animals, 1991,Vol.4,No.4:297-302
58. Emanuele M.,Gianfranco p. chemical composition and nutritional properties of commercial products of mare powder. Journal of food composition and analysis. 1998,11:178-187
59. Valieva,T.A.;Valiev,A.G. Lipid and fatty acid contents, and lipid peroxidation in freeze-dries mare milk during prolonged storage with antioxidants. Voprosy Pitaniya, 1993,No.5:61-65
60. Valiev, A.G.;Valieva,T.A.etc.the influence of essential fatty acids of mare's milk on the immune system and nonspecific resistance of rats. Voprosy Pitaniya, 1999,Vol.68,No.3:3-6
61. Prateek jain; Gupta, S.K.nutritional potentiality of mares'milk. Indian dairyman, 1998,Vol.50,No.8:39-42
62. Fantova,M.; kukanova,J. Possibilities of utilization of mares' milk. Agriculture tropica et subtropica, 1998,No.31:177-181
63. FrancescoB.StefaniaL.E.Thermal sensitivity of mares'milk proteins. Journal of dairy research. 1994,61:419-422
64. Lozovich,S. Medical uses of whole and fermented mare milk in Russia. Cultured Dairy Products Journal, 1995,Vol.30,No. 1:18-19
65. Szakaly, S.Role of fermented milk products in protection of human health. Tejgazdasag, 1999, Vol59,No.2:15-18
66. Y. Tamime. special feature-kefir, koumiss and kishk. Dairy industries international. 1999, Vol. 64,No. 5:32-33
67. Mistry, V.V. Fermented milks and cream. Applied dairy microliology, 2001,No.Ed.2:301-325
68. Prehrambene, J.L.Samarzija,D. Nutritional, health and technological properties of cultured products.Mljekarstvo, 1996,Vol.46,No.4:265-273
69. Storch G, Composition and properties of mares' milk and koumiss,with particular regard to dietetic aspects. Thesis,Justus-Liebig-Universitat Giessen,German,Federal Republic 1985,98
70. Omel'chenko,N.P. Nutrition of breast-feeding women in Yakut conditions. Voprosy Pitaniya, 1993,No. 1:23-27
71. Montanari,G.Zambonelli,C.Koumiss,a cultured milk drink.lndustrie Alimentari, 1997,Vol.36,No.355:5-9
72. Molochnikov, V.V.;Anisimov, S.V. Mixture for koumiss and manufacturing method. USSR,Patent, 1991,No. SU 1 664 238
73. Pastukhova Z M, Gerbeda V V. Conparative composition of lipids of mares'milk and of a koumiss mixture based on cows' milk. FSTA, 1982
74. Robinson, R.K.; Tamime,A.Y. Microbiology of fermented milks. Dairy microbiology handbook:the microbiology of milk and milk products,2002,No.Ed:367-430
75. Ishii,S. Survey of the koumiss of Sakha. Milk Science, 1999, Vol48,No.2:193-195
76. Gregurek,L.Dairy cultures of microorganisms in the manufacture of cultured milks. Mljekarstvo, 1997,Vol.47,No.2:103-113
77. Ishii, S.; Kikuchi, M.; Takao, S. Isolation and identification of lactic acid bacteria and yeasts from "Chigo" in Inner Mongolia, China. Animal Science and Technology, 1997,Vol. 68,No. 3:325-329
78. Montanari ,G.; Zambonelli, C.; Grazia, L. Saccharomyces unisporus as the principal alcohol fermentation microorganism of traditional koumiss, etal. J. of Dairy Research, 1996, Vol.63,No. 2:327-331
79. Ham, J. S.; Jeong, J. Y. etal. Effects of feeding with Lactobacillus plantarum and Candida Kefyr isolated from Mongoiian koumiss on the growth and fecal microflora of broilers. Korean Journal of Dairy Science ,1999,Vol. 21,No. 3:241-246
80. Ham,J.S.etl.fermentation of goat milk using Lactobacillus plantarurn and Candida Kefyr isolated from Mongolian koumiss. Korean Journal of Dairy Science, 1999,Vol. 21,No. 3:247-254
81. Ishii,S. Kikuchi, M.; Takao, S.Identification of compounds causing symbiotic growth of Lactobacillus paracasei subsp. Tolerans and kluyveromyces marxianus var. lactis in chigo, inner Mongolia, china. Animal science Journal, 1999,Vol.70,No.2:81-89
82. Hiroshi shimizu,taiji mizuguchi. Nisin production by mixed-culture systerm consisting of Lactococcus lactis and Kluyveromyces marxianus. Applied and environmental microbiology, 1999,4:3134-3141
83. Jain, A.k.; Prajapati,j.B. Associative growth in dairy starter cultures, jounal of dairying,foods and home sciences, 1998,Vol. 17,No. 1:1-11
84. Lukerchenko,V.N. A new strain of yeast for lactose fermentation. Pishchevaya Promyshlennost, 2001,No.2:58
85. Libudzisz, Z.intestinal microflora and new generations of fermented milk beverages.
Zywienie Czlowieka I Metabolizm, 1999,Vol.26,No. Suppl:5-13
86. Gladkova,E. How to organize koumiss production on the farm. Konevodstvo I Konnyisport, 1997,No.4:8-9
87. Marshall,V.M. starter cultures for milk fermentation and their characteristics. Journal of the society of dairy technology, 1993,Vol.46,No.2:49-56
88. Fleet, G.H.Yeasts in dairy products.The Journal of Applied Bacteriology, 1990,68:199-121
89. Graciela L. Garrote,Analia G. Inhibitory power of kefir: the role of organic acid. J. of food protection. 2000, Vol 63,No.3:364-369
90. Helander, I.M.,A.Von Wright, and T.M. Potential of lactic acid bacteria and novel antimicrobials against gram-negative bacteria Mattila-Sandholm. Trends Food Sci. Technol. 1997,8:146-150
91. Presser. K.A.,D.A. Ratkowsky. and T. Ross. Modlling the growth rate of Escherichia coli as a function of pH and lactic acid concentration. Appl. Environ. Microbiol, 1997,63: 2355-2360