A kind of composite bacterial agent based on bacillus subtilis bs1 and its application

A technology of Bacillus subtilis and compound inoculum, which is applied to the compound inoculum based on Bacillus subtilis BS1 and its application field, and achieves the effects of obvious reduction, increasing compost temperature and shortening time.

Active Publication Date: 2021-02-09
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacillus is the dominant flora in the compost. The Bacillus subtilis isolated from the compost in the present invention has strong protease, amylase and cellulase activities, is sensitive to various antibiotics, and has a strong effect on ammonium nitrogen. The ability to use it, screen it for the development of composting compound bacteria agent, and obtain good application effect, especially have a certain reduction effect on antibiotic resistance genes and pathogenic bacteria in composting. Currently, there is no report on this aspect of research.

Method used

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  • A kind of composite bacterial agent based on bacillus subtilis bs1 and its application
  • A kind of composite bacterial agent based on bacillus subtilis bs1 and its application
  • A kind of composite bacterial agent based on bacillus subtilis bs1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Obtaining of Bacillus subtilis BS1:

[0041] It is isolated and purified from pig manure compost, and identified as Bacillus subtilis according to its physiological and biochemical indicators and sequence analysis. This strain has amylase, protease and cellulase activities; it is sensitive to many different types of antibiotics, sensitive antibiotics include: tetracycline , chloramphenicol, erythromycin, norfloxacin, kanamycin, co-trimoxazole; and can use a variety of nitrogen sources, including (NH 4 ) 2 SO 4 , CO(NH 2 ) 2 , NH 4 Cl, KNO 3 、NaNO 2 , NH 3 ·H 2 O.

[0042] (1) Colony morphological characteristics:

[0043] Round or irregular shape, rough surface, opaque, dirty white ( figure 1 ).

[0044] (2) Culture characteristics

[0045] Bacillus subtilis can survive in basal medium LB (sodium chloride, tryptone, yeast extract, adjust pH to 7.0 with 5mol / L NaOH, 121°C, sterilize for 20min), and its optimum growth temperature is 30°C- 37°C. Colony growt...

Embodiment 2

[0050] Biological characteristics of Bacillus subtilis BS1:

[0051] Using three methods (disc, single colony spot, punching) to explore the antibacterial test of BS1 against Salmonella, Pasteurella, and Streptococcus in vitro, no antibacterial zone was found, BS1 has a fast propagation speed and large colonies, and it may pass Competing for nutrients and ecological sites and other means to win development space for itself, squeeze the living environment of pathogenic microorganisms, so as to achieve the purpose of reducing pathogenic bacteria. The sensitivity test of the BS1 strain to antibiotics showed that the diameter of the inhibition zone was greater than 15mm, and it was sensitive to multiple antibiotics, indicating that the strain did not carry antibiotic resistance genes and would not increase the horizontal transfer of resistance genes.

[0052] Through enzyme activity test, it was found that BS1 had higher protease activity, amylase activity and cellulase activity. ...

Embodiment 3

[0055] A compound microbial agent, after mixing Bacillus subtilis, Bacillus licheniformis, Bacillus megaterium and Saccharomyces cerevisiae at a volume ratio of 1L:1L:1L:2L, centrifuge and resuspend in 0.5L sterilized LB liquid medium Prepared; Used in the following examples;

[0056] Before mixing, the effective bacterium concentration of described Bacillus subtilis is 8x10 8 CFU / mL, the effective bacterial concentration of Bacillus licheniformis is 1.7x10 8 CFU / mL; effective bacterial concentration of Bacillus megaterium 1.18x10 8 CFU / mL; the effective bacterial concentration of Saccharomyces cerevisiae is 6.2x10 7 CFU / mL;

[0057] The Bacillus subtilis is Bacillus subtilis BS1, and the preservation number is: CCTCC NO: M 2019185; the Bacillus licheniformis is Bacillus licheniformis L3; the Bacillus megaterium is Bacillus megaterium C2; Yeast Y2.

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Abstract

The invention belongs to the field of microbes and their applications, and specifically discloses a composite bacterial agent based on Bacillus subtilis BS1 and its application. The Bacillus subtilis has been preserved in the China Center for Type Culture Collection, and the preservation number is: CCTCC NO: M 2019185. The compound microbial agent prepared by the bacterial strain and Saccharomyces cerevisiae, Bacillus megaterium and Bacillus licheniformis can be used for composting livestock and poultry manure. The composite microbial agent of the invention can increase the composting temperature in composting, shorten the time to reach high temperature, deodorize and fix nitrogen, and has a significant reduction effect on various antibiotic resistance genes and pathogenic bacteria in composting.

Description

technical field [0001] The invention belongs to the field of microbes and their applications, and in particular relates to a composite bacterial agent based on bacillus subtilis BS1 and its application; the composite bacterial agent can be used for livestock and poultry manure composting. Background technique [0002] With the rapid development of my country's breeding industry, livestock and poultry manure pollution has become one of the sources of agricultural non-point source pollution. The large amount of livestock and poultry manure discharged and the difficulty of treatment not only seriously affected the health and production performance of livestock and poultry, but also caused huge pressure on the ecological environment such as the surrounding water body and soil. At the same time, antibiotic resistance genes (ARGs) carried by livestock and poultry manure, as a new type of pollutant, can spread in the environment and are difficult to control and eliminate. These fa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/18C05F17/20C05F17/50C05G3/00C05G3/60C05F15/00C05F11/08C12R1/125C12R1/10C12R1/11C12R1/865
CPCC05F3/00C05F11/08C05F17/00C05G3/00C12N1/18C12N1/20Y02W30/40
Inventor 吴健曹蕊李珂莫尚昆任竹青
Owner HUAZHONG AGRI UNIV
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