草鱼铜绿假单胞菌灭活疫苗发酵培养基的优化及其在发酵罐的生长试验
|
摘要
为培养优质的铜绿假单胞菌Pseudomonas aeruginosa菌液,通过单因素试验对草鱼铜绿假单胞菌灭活疫苗发酵培养基的氮源、碳源和磷酸盐成分进行了筛选,采用正交试验法对培养基各主要成分的用量进行了优化组合,并经过验证试验绘制出了铜绿假单胞菌JP802在优化培养基条件下的5 L发酵罐生长曲线。结果表明:草鱼赤皮病铜绿假单胞菌JP802发酵培养基中最佳氮源为蛋白胨+牛肉膏+酵母膏,最佳碳源为葡萄糖,最佳磷酸盐为磷酸氢二钾;确立了培养基的优化配方为蛋白胨10 g/L、牛肉膏5.0 g/L、酵母膏2.5 g/L、葡萄糖5.0 g/L、磷酸氢二钾0.75 g/L、氯化钠5.0 g/L,JP802菌株在此培养基中发酵14 h菌体浓度达到最大(OD_(600 nm)值为6.44)。研究表明,通过对发酵培养基的优化,可以获得更高产量的铜绿假单胞菌JP802发酵菌液。
The nitrogen source, carbon source and phosphate were screened in culture medium of Pseudomonas aeruginosa inactivated vaccine in a single-factor test, and different ingredients of the medium were optimized in an orthogonal experiment, with a verification test of growth of the bacterium in a 5 L fermentor. It was found that the best fermentation of Pseudomonas aeruginosa in grass carp Ctenopharyngodon idellus with red skin disease was observed under the conditions of peptone+beef extract+yeast extract as nitrogen source, glucose as carbon source, K_2PHO_4 as phosphate source, at a dose of peptone 10 g/L, beef extract 5.0 g/L, yeast extract 2.5 g/L, glucose 5.0 g/L, K_2HPO_4 0.75 g/L, and NaCl 5.0 g/L. The fermentation kinetics analysis revealed that the stain had the maximal cell density at 6.44 of OD_(600 nm) in 14 h culture, indicating that a high yield of Pseudomonas aeruginosa JP802 fermentation broth is obtained by the optimized fermentation medium.
The nitrogen source, carbon source and phosphate were screened in culture medium of Pseudomonas aeruginosa inactivated vaccine in a single-factor test, and different ingredients of the medium were optimized in an orthogonal experiment, with a verification test of growth of the bacterium in a 5 L fermentor. It was found that the best fermentation of Pseudomonas aeruginosa in grass carp Ctenopharyngodon idellus with red skin disease was observed under the conditions of peptone+beef extract+yeast extract as nitrogen source, glucose as carbon source, K_2PHO_4 as phosphate source, at a dose of peptone 10 g/L, beef extract 5.0 g/L, yeast extract 2.5 g/L, glucose 5.0 g/L, K_2HPO_4 0.75 g/L, and NaCl 5.0 g/L. The fermentation kinetics analysis revealed that the stain had the maximal cell density at 6.44 of OD_(600 nm) in 14 h culture, indicating that a high yield of Pseudomonas aeruginosa JP802 fermentation broth is obtained by the optimized fermentation medium.
引文
[1] 霍瑞清,贾海琴,唐丽.2013年至2015年铜绿假单胞菌医院感染分布及耐药性分析[J].内蒙古医学杂志,2017,49(1):23-25.
[2] 陶永红,杨茂生.养殖场铜绿假单胞菌污染的检测报告[J].贵州畜牧兽医,2013,37(3):6-8.
[3] 牛钟相,李雅林.动物绿脓杆菌病研究进展[J].动物医学进展,2003,24(1):16-18.
[4] 郭政,赵庆中,张先锋.长江江豚感染铜绿假单胞菌肺炎的诊治[J].兽类学报,2007,27(1):86-91.
[5] 于道平,糜励,蒋文华,等.长江江豚慢性皮肤炎症的初报[J].浙江海洋学院学报:自然科学版,2005,24(4):335-338.
[6] 刘振国,董金海,李鲁,等.宽吻海豚急性铜绿假单胞菌感染的治疗研究[J].中国水产科学,2000,7(2):123-125.
[7] 崔来宾,叶星,邓国成,等.草鱼铜绿假单胞菌的鉴定及药物敏感性分析[J].大连海洋大学学报,2010,25(6):488-494.
[8] 高岩,裴超,张超,等.草鱼呼肠孤病毒疫苗的研发与应用[J].水产科学,2017,36(2):237-242.
[9] 吴淑勤,陶家发,巩华,等.渔用疫苗发展现状及趋势[J].中国渔业质量与标准,2014,4(1):1-13.
[10] Sommerset I,Kross?y B,Biering E,et al.Vaccines for fish in aquaculture[J].Expert Review of Vaccines,2005,4(1):89-101.
[11] Str?m-Bestor M,Mustam?ki N,Heinikainen S,et al.Introduction of Yersinia ruckeri biotype 2 into Finnish fish farms[J].Aquaculture,2010,308(1-2):1-5.
[12] Brudeseth B E,Wiulsr?d R,Fredriksen B N,et al.Status and future perspectives of vaccines for industrialised fin—fish farming[J].Fish & Shellfish Immunology,2013,35(6):1759-1768.
[13] 李钊,蒋捍东.培养基对不同产酶铜绿假单胞生物膜菌敏感性的影响[J].山东医药,2008,48(13):4-6.
[14] 张秀明,张文生,李慧,等.培养条件对乳酸乳球菌N302合成乳酸链球菌素的影响[J].南开大学学报:自然科学版,2005,38(3):88-92.
[15] 卢金珍,赵为,任俊.乳链菌肽(Nisin)发酵培养基的优化[J].食品研究与开发,2010,31(4):122-126.
[16] 黄秀敏,李学优,叶俊豪,等.1株益生蜡样芽孢杆菌发酵培养基优化研究[J].安徽农业科学,2016,44(30):7-8,15.
[17] 夏立群,王蓓,夏洪丽,等.鰤鱼诺卡氏菌培养条件及培养基的优化[J].南方水产科学,2013,9(3):51-56.
[18] 黄秀敏,张宜靖,李学优,等.植物乳杆菌发酵培养基的优化[J].安徽农业科学,2016,44(24):6-7,22.
[19] 周德庆.微生物学教程[M].3版.北京:高等教育出版社,2011:87-91.
[20] 阙斐.枯草芽孢杆菌液体发酵培养基成分优化研究[J].广州化工,2017,45(18):68-70.
[2] 陶永红,杨茂生.养殖场铜绿假单胞菌污染的检测报告[J].贵州畜牧兽医,2013,37(3):6-8.
[3] 牛钟相,李雅林.动物绿脓杆菌病研究进展[J].动物医学进展,2003,24(1):16-18.
[4] 郭政,赵庆中,张先锋.长江江豚感染铜绿假单胞菌肺炎的诊治[J].兽类学报,2007,27(1):86-91.
[5] 于道平,糜励,蒋文华,等.长江江豚慢性皮肤炎症的初报[J].浙江海洋学院学报:自然科学版,2005,24(4):335-338.
[6] 刘振国,董金海,李鲁,等.宽吻海豚急性铜绿假单胞菌感染的治疗研究[J].中国水产科学,2000,7(2):123-125.
[7] 崔来宾,叶星,邓国成,等.草鱼铜绿假单胞菌的鉴定及药物敏感性分析[J].大连海洋大学学报,2010,25(6):488-494.
[8] 高岩,裴超,张超,等.草鱼呼肠孤病毒疫苗的研发与应用[J].水产科学,2017,36(2):237-242.
[9] 吴淑勤,陶家发,巩华,等.渔用疫苗发展现状及趋势[J].中国渔业质量与标准,2014,4(1):1-13.
[10] Sommerset I,Kross?y B,Biering E,et al.Vaccines for fish in aquaculture[J].Expert Review of Vaccines,2005,4(1):89-101.
[11] Str?m-Bestor M,Mustam?ki N,Heinikainen S,et al.Introduction of Yersinia ruckeri biotype 2 into Finnish fish farms[J].Aquaculture,2010,308(1-2):1-5.
[12] Brudeseth B E,Wiulsr?d R,Fredriksen B N,et al.Status and future perspectives of vaccines for industrialised fin—fish farming[J].Fish & Shellfish Immunology,2013,35(6):1759-1768.
[13] 李钊,蒋捍东.培养基对不同产酶铜绿假单胞生物膜菌敏感性的影响[J].山东医药,2008,48(13):4-6.
[14] 张秀明,张文生,李慧,等.培养条件对乳酸乳球菌N302合成乳酸链球菌素的影响[J].南开大学学报:自然科学版,2005,38(3):88-92.
[15] 卢金珍,赵为,任俊.乳链菌肽(Nisin)发酵培养基的优化[J].食品研究与开发,2010,31(4):122-126.
[16] 黄秀敏,李学优,叶俊豪,等.1株益生蜡样芽孢杆菌发酵培养基优化研究[J].安徽农业科学,2016,44(30):7-8,15.
[17] 夏立群,王蓓,夏洪丽,等.鰤鱼诺卡氏菌培养条件及培养基的优化[J].南方水产科学,2013,9(3):51-56.
[18] 黄秀敏,张宜靖,李学优,等.植物乳杆菌发酵培养基的优化[J].安徽农业科学,2016,44(24):6-7,22.
[19] 周德庆.微生物学教程[M].3版.北京:高等教育出版社,2011:87-91.
[20] 阙斐.枯草芽孢杆菌液体发酵培养基成分优化研究[J].广州化工,2017,45(18):68-70.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |