Production method of bacillus amyloliquefaciens antimicrobial lipopeptide and application thereof in prawn feed

Amyloliquefaciens technology and a production method are applied in the production of Bacillus amyloliquefaciens antibacterial lipopeptide and its application in shrimp feed, can solve the problems of restricting the large-scale use of feed, narrow antibacterial spectrum, etc., and improve the utilization of feed rate, growth promotion and immune regulation, the effect of reducing production costs

Inactive Publication Date: 2014-01-01
NANJING AGRICULTURAL UNIVERSITY +2
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Problems solved by technology

However, animal-derived antimicrobial substances have a narrow antibacterial spectrum an

Method used

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  • Production method of bacillus amyloliquefaciens antimicrobial lipopeptide and application thereof in prawn feed

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Embodiment 1

[0028] Bacillus amyloliquefaciens ( Bacillus amyloliquefaciens ) fmb-50 is Bacillus amyloliquefaciens ES-2-4 via N + The high-yielding mutant strains bred by ion mutagenesis, the high-yield mutant strains bred by nitrosoguanidine mutagenesis, and other high-yielding mutant strains obtained by protoplast fusion were named fmb-50. B. amyloliquefaciens fmb-50 can obtain the highest antibacterial lipopeptide production with corn flour as the carbon source, and its production is 3.4 times that of the parent strain ES-2-4. [Yao Shulin, Lu Zhaoxin et al., Taguchi method optimization B. amyloliquefaciens fmb-50 surfactin-producing industrial fermentation medium, Journal of Beijing University of Chemical Technology, 2012,39(4):77-83]. Original strain of fmb-50 B. amyloliquefaciens ES-2-4, B. subtilis 168 and B. subtilis fmbJ (the strains are all known and public, see Huang Xianqing, etc., the effect of Bacillus subtilis fmbJ lipopeptide inhibition point Penicillium and ...

Embodiment 2

[0036] The activated pathogenic bacteria Vibrio parahaemolyticus, Vibrio harveyi, Vibrio alginolyticus, Streptococcus agalactiae (the strains are all known and public, see Chen Jixiang et al., Pathogenic Vibrio harveyi hemolytic Gene cloning and detection, 2005, Chinese Fisheries Science; Zhai Xiumei etc. Effects of Vibrio parahaemolyticus on physiological and biochemical indicators of Penaeus vannamei, 2007, Journal of Shanghai Fisheries University; Yan Qingpi et al., Effects of different environmental conditions on the adhesion of Vibrio alginolyticus to intestinal mucus of large yellow croaker, 2006, Acta Aquatica Sinica; Wu Yingrui et al., Research on the inhibitory activity of 153 kinds of Chinese herbal medicines against Streptococcus agalactiae and Streptococcus iniae in tilapia, Journal of Northwest A&F University, 2013) respectively received in TSB (tryptone soybean liquid medium), cultured at 36±1℃ for 4 ~6 h, adjust bacterial OD with TSB medium 600 value, to a ...

Embodiment 3

[0046] Litopenaeus vannamei with normal appearance, healthy body and average tail weight of 1.02 ± 0.10 g were selected as experimental shrimps. Fish meal, soybean meal, peanut meal and brewer's yeast were used as protein sources, fish oil and soybean oil (1:1) as fat source, flour as carbon source, and appropriate minerals and vitamins were added as compound feed for Litopenaeus vannamei (Table 2 ). According to adding 6g (A group), 8g (B group), 10g (C group) described antimicrobial substance feed additive (preparation in the first method in embodiment 1, potency 40000 IU / g) in every kilogram of compound feed, CK group without antibacterial substances (Supplementary Table 2). Three repetitions were set up for each treatment, and 30 Litopenaeus vannamei were stocked in each glass steel bucket, and the water capacity of the glass steel bucket was 0.3 m 3 . The water temperature was (27. 5 ± 1. 5) ℃, the seawater specific gravity was 1. 012 ± 0. 002, and the breeding experi...

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Abstract

The invention relates to a production method of bacillus amyloliquefaciens antimicrobial lipopeptide and application thereof in prawn feeds and belongs to the technical fields of aquiculture and biology in the field of agriculture. The production method comprises the following steps that bacillus amyloliquefaciens fmb-50 with the collection number of CGMCC No.6249 is used as a strain and is sequentially subjected to seed culture, amplification culture and fermentation production to obtain fermentation liquor; the thalli of the fermentation liquor are removed through microfiltration or membrane filtration and nanofiltration; corn starch or porous starch is added for absorption and spray drying is carried out, thus obtaining an antibacterial substance feed additive with the antibacterial valence of 40000-50000 IU/g or 250000 IU/g. The antibacterial substance which is produced from the bacillus amyloliquefaciens fmb-50 through fermentation can be industrially produced in a large scale; the production period is short and production cost is greatly reduced; when used in the prawn feeds, the antibacterial substance can reduce the mortality rate, improve the utilization ratio of the prawn feeds, promote the nonspecific immunoreaction of prawns and enhance the energy metabolism level.

Description

[0001] technical field [0002] The invention belongs to the technical field of aquaculture and the field of biotechnology in the agricultural field, and relates to a production method of bacillus amyloliquefaciens antibacterial lipopeptide and its application in prawn feed. Background technique [0003] Antibiotics are used in the aquaculture industry during production and processing, mainly to prevent (preventatives) and treat (therapeutics) bacterial diseases. Adding antibiotics to feed can prevent aquatic diseases, increase aquatic production and save feed. However, the frequent use of antibiotics will lead to the formation of drug resistance in the bacterial flora in the breeding environment. Once the highly pathogenic bacteria acquire drug resistance to the antibiotics shared by humans and animals, the efficacy of the effective antibiotics may be reduced after human infection. Little or even ineffective. In addition, antibiotics left in aquatic products enter the hum...

Claims

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Application Information

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IPC IPC(8): A23K1/18A23K1/17A23K20/195
Inventor 陆兆新赵威钢施雨含吕凤霞别小妹张充
Owner NANJING AGRICULTURAL UNIVERSITY
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