Prawn intestinal enterococcus strain as well as screening method and application thereof

A technology of enterococci and strains, which is applied in the field of Enterococcus strains and their screening in shrimp, can solve the problems of complicated types of lactic acid bacteria preparations and difficult verification of active ingredients, and achieve the effect of improving growth and disease resistance

Active Publication Date: 2019-04-26
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many types of lactic acid bacteria preparations for aquatic products on the market, and the active

Method used

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  • Prawn intestinal enterococcus strain as well as screening method and application thereof
  • Prawn intestinal enterococcus strain as well as screening method and application thereof
  • Prawn intestinal enterococcus strain as well as screening method and application thereof

Examples

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Embodiment 1

[0024] 1. Isolation and purification of bacterial strain of the present invention

[0025] On a sterile ultra-clean bench, use a sterilized toothpick to pick out the intestines of live shrimp and put them into a sterilized 2 ml EP tube, add 1.5 ml of autoclaved PBS buffer solution (pH 7.0), vibrate vigorously for 30 s, and place Incubate on a shaker at 28°C and 180 rpm for 30 min; take an appropriate amount of culture medium for serial dilution (10 -2 、10 -3 、10 -4 、10 -5 ), spread evenly on the MRS solid medium, and culture it upside down at 28°C for 2-3 days; pick a single colony with a sterile inoculation loop, continue to purify and culture it on the MRS plate for 2 times, and then add 20% sterile glycerol to preserve Seed at -80°C.

[0026] 2. the bacteriostasis of bacterial strain of the present invention

[0027] After activating the isolated strain, use MRS to culture overnight at 28°C, then measure OD 600 And adjust its OD value to 0.6-0.8 for use. The indicato...

Embodiment 2

[0035] This experiment was carried out at the Yongxing base of the Wenzhou Marine Aquaculture Research Institute in Zhejiang Province. A total of 10 600L (diameter 1m, height 0.8m) Litopenaeus vannamei breeding ecological experiment barrels were set up, of which 5 barrels were used as controls and 5 barrels were used as Enterococcus NBU1 addition group. Shrimp seedlings were placed uniformly, and 210 shrimps were placed in each experimental bucket (average body length was 7.08±0.33 cm, average body weight was 4.34±0.64 g), and the stocking density was 500 shrimps / m 3 , The experiment was officially started 2 days after the seedlings were released. Bacterial agent activation and addition: Take the preserved strains and activate them on the MRS solid medium. After a single colony grows, pick a single colony and culture it in 10 mL sterile MRS broth medium overnight, and then add 2% (V / V) was transferred to fresh MRS broth medium for 24 h, and the bacterial concentration reache...

Embodiment 3

[0037] This experiment was completed in Xianxiang Aquaculture Base, using a standardized breeding tank (600L) system, which is equipped with consistent oxygenation, water replacement and other devices. Select sick shrimp seedlings with a size of 2-3 cm, about 200 tails per barrel (to ensure that the total weight added is consistent). The control group and the NBU1 addition group were respectively set up, and the addition method was as follows: Enterococcus NBU1 was cultured with MRS medium to the OD of the bacterial concentration 600 When the absorbance value is about 1.0, take 20g of sucrose and add 200 ml of bacterial liquid, then take 5% of the total bacterial liquid to stir the feed, ferment for 1 hour and then feed, and the remaining 95% of the bacterial liquid is activated and sprinkled into the breeding tank. The solution was replaced with sterile medium. Set 5 repetitions for each group. Feed 3 times a day (7:00, 12:00, 17:00), and the initial daily feeding amount ac...

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Abstract

The invention discloses a prawn intestinal enterococcus strain as well as a screening method and application thereof. The prawn intestinal enterococcus strain is characterized in that the strain is NBU1 strain which is classified and named as Enterococcus sp., and was preserved in China General Microbiological Culture Collection Center on November 12, 2018, with a preservation number of CGMCC No.16735; the application of the strain in preparing a staphylococcus aureus inhibitor and/or a vibrio anguillarum inhibitor as well as preparing a probiotic additive for prawn culture has the advantagesthat better inhibitory effects on staphylococcus aureus and vibrio anguillarum are realized; the yield and the survival rate of prawns and the conversion rate of feed can be improved after the strainis fermented with the feed; in addition, the yield and the survival rate of sick prawns can be improved; the prawn intestinal enterococcus strain can be used as a high-efficiency green feed additive and an antibiotic substitute for aquatic animal cultivation.

Description

technical field [0001] The invention relates to a strain of enterococcus and application thereof, in particular to a strain of enterococcus prawn enterococcus and a screening method and application thereof. Background technique [0002] Shrimp farming has always been one of the important supporting industries of aquaculture in my country. However, with the improvement of the intensive degree of prawn farming, the frequent occurrence of prawn diseases has greatly restricted the development of the prawn farming industry in our city. At present, there is no effective and rapid method to prevent and treat shrimp diseases. People usually use antibiotics and chemically synthesized drugs to prevent and control aquatic animal diseases. However, the long-term and large-scale use of antibiotics will easily lead to the generation of drug resistance of Penaeus vannamei, the decline of its own immune function, and the variation of pathogenic bacteria, resulting in many drug-resistant st...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A61K35/744A61P31/04C12R1/01
CPCA61K2035/115A61P31/04C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 郭海朋张德民陈伟鲍方剑杜世聪
Owner NINGBO UNIV
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