Rainbow trout-derived bacillus subtilis strain as well as screening method and application thereof

A technology derived from Bacillus subtilis and rainbow trout, applied in the field of microorganisms, can solve problems such as inability to obtain better effects and promotion, and achieve the improvement of immunity and digestion-related enzyme activities in rainbow trout-related tissues, improvement of anti-infection ability, and promotion of body length. Effect

Active Publication Date: 2020-09-01
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Due to the differences in germplasm, habitat and feeding conditions, the practical application of foreign research and development and commercia

Method used

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  • Rainbow trout-derived bacillus subtilis strain as well as screening method and application thereof
  • Rainbow trout-derived bacillus subtilis strain as well as screening method and application thereof
  • Rainbow trout-derived bacillus subtilis strain as well as screening method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Screening and identification of Bacillus subtilis strain RT-BS07

[0043] 1. Isolation and purification of RT-BS07 strain

[0044]Representative rainbow trout farms in Qinghai, Liaoning and other cold-water fish main breeding areas were selected as sampling sites, and the probiotics in the intestinal tract of rainbow trout were initially screened and isolated. Under aseptic conditions, the body surface of rainbow trout was disinfected, the intestinal tissue of rainbow trout was dissected, washed three times with sterile phosphate buffered saline (PBS), the intestinal tissue was broken with a tissue homogenizer to obtain a tissue homogenate, and the tissue was homogenized. The slurry was collected in 1 mL of sterile saline, and after serial dilution, 100 μL was spread on the surface of the selective medium, and placed in a constant temperature incubator at 28°C for upside-down culture. Pick a single colony on the plate and perform pure culture by streak cultur...

Embodiment 2

[0063] Example 2 Analysis of biological characteristics of Bacillus subtilis strain RT-BS07

[0064] 1. Analysis of antibacterial properties of RT-BS07 strain in vitro

[0065] Polymyxin B was used as positive control, PBS was used as negative control, and common disease microorganisms in the breeding process of rainbow trout were selected, and the effects of RT-BS07 strain on Aeromonas salmonicida (Aeromonas salmonicida), Pseudomonas fluorescens (Pseudomonas fluorescens), A.sobria, A.veronii, Yersinia ruckeri, and A.hydrophila (For specific information on the strains used, see: Zhang Xiao, Li Shaowu, Wang Di, Cao Yongsheng, Lu Tongyan. Establishment and application of qPCR rapid detection method for Yersinia ruckerii [J]. Chinese Fisheries Science, 2017, 24(6) :1254-1260.) Antagonistic antibacterial effect.

[0066] The specific measurement method is: add the selected representative pathogenic microorganisms to the solid medium and pour the plate, punch holes in the Oxford ...

Embodiment 3

[0099] Example 3 Evaluation of the safety and application effect of RT-BS07 strain on rainbow trout

[0100] 1. Preparation of bait containing probiotic RT-BS07 strain and immune feeding of experimental fish

[0101] Take out the RT-BS07 strain frozen at -80°C, and inoculate 5 μL of the frozen bacteria into 5 mL of LB liquid medium (yeast extract 5 g; tryptone 10 g; sodium chloride 10 g; adjust the pH to 7.0 with NaOH, Add water to make up to 1L, keep at 121°C for 15min (autoclave), shake the bacteria at 28°C at 120 rpm for 12h to obtain a culture solution, then take 2mL of the culture solution and add it to 2L liquid LB medium, continue for 120 rpm Shake the bacteria per minute for 18 hours to prepare the bacterial suspension.

[0102] Aile silver fish feed was selected as the basic feed, and a feed with a diameter of 2 mm was selected according to the size of the experimental fish. The active ingredients of the feed included: 45% crude protein, 20% crude fat, 17.9% nitrogen...

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Abstract

The invention relates to a rainbow trout-derived bacillus subtilis strain and a screening method and application thereof. The screening method comprises following steps: at first, a bacillus subtilisRT-BS07 strain is screened and identified from a healthy rainbow trout intestine, the biological characteristics of the strain are analyzed, and the probiotic functions of the strain in the aspects ofgrowth promotion, infection resistance, and the like of rainbow trout are evaluated through in-vivo tests. The obtained RT-BS07 strain has the characteristics of safety, acid resistance, salt resistance, capability of well adhering to the intestinal tract of a host and capability of inhibiting the growth of pathogenic bacteria; the increase of the body length, the body mass, the spleen weight andthe intestinal tract length of rainbow trout can be effectively promoted; the activity of immune and digestion related enzymes in rainbow trout related tissues can be effectively improved; the up-regulated expression of growth and immunity related genes in rainbow trout tissues is promoted; the anti-infection performance of rainbow trout is improved; and after rainbow trout is attacked and infected by aeromonas hydrophila, the death rate of an experimental group is lower than that of a control group by 30%.

Description

technical field [0001] The invention relates to a bacillus subtilis strain derived from rainbow trout and a screening method and application thereof, belonging to the technical field of microorganisms. Background technique [0002] Probiotics are derived from the Greek word Probiotics, which means "symbiosis", which means "playing a complementary role in the life maintenance of animals", which is opposite to Antibiotics (antibiotics). Specifically, probiotics are a type of active microorganisms that are beneficial to the host by colonizing in the organism and changing the composition of the flora in a certain part of the host. By regulating the immune function of the host mucosa and system or by regulating the balance of intestinal flora, it promotes nutrient absorption and maintains intestinal health, thereby producing single microorganisms or mixed microorganisms with a clear composition that are beneficial to the health of the host. [0003] Aquatic probiotics can enhanc...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/02A23K50/80A23K10/18A61K35/742A61P31/04C12R1/125
CPCC12N1/02A23K50/80A23K10/18A61K35/742A61P31/04C12R2001/125C12N1/205
Inventor 王荻李绍戊曹永生卢彤岩刘红柏
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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