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Preparation method and application of Brevibacterium cold-resistant zl-2 and its composting low-temperature starter bacterial agent

A cold-resistant Brevibacterium, ZL-2 technology, applied in the biological field, can solve the problems of difficult rapid heating and start-up of piles, and ineffective fermentation of manure.

Active Publication Date: 2022-03-01
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, compost is easily affected by the ambient temperature. When the ambient temperature is low, it is difficult for the compost to heat up quickly and start, resulting in the ineffective fermentation of manure.

Method used

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  • Preparation method and application of Brevibacterium cold-resistant zl-2 and its composting low-temperature starter bacterial agent
  • Preparation method and application of Brevibacterium cold-resistant zl-2 and its composting low-temperature starter bacterial agent
  • Preparation method and application of Brevibacterium cold-resistant zl-2 and its composting low-temperature starter bacterial agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Isolation, screening and identification of Brevibacterium cold-resistant ZL-2

[0031] The bacterial strain of the present invention is isolated from the winter permafrost on the campus of Dalian University of Technology.

[0032] 1. Isolation and screening of Brevibacterium cold-resistant ZL-2

[0033] (1) Weigh 10 g of collected permafrost soil samples and add them to a conical flask containing 90 mL of PBS buffer solution, and add sterile glass beads (which help the sample to disperse evenly in PBS buffer solution). , 180rpm shaking table vibration 20min, leave standstill 10min, prepare permafrost soil bacteria suspension (being 10 -1 permafrost soil dilution); then carry out 10-fold gradient dilution with PBS buffer solution to obtain 10 -2 、10 -3 , 10 -4 、10 -5 、10 -6 、10 -7 、10 -8 Permafrost soil dilution suspension, choose 10 -4 、10 -5 、10 -6 、10 -7 , 10 -8 100 μL each of the diluted suspension was evenly spread on the beef extract peptone...

Embodiment 2

[0047] The preparation method of Brevibacterium cold-resistant ZL-2 composting low-temperature starting bacterial agent, the specific steps are: the number of effective viable bacteria>1×10 8 Mix CFU / mL Brevibacterium cold-tolerant ZL-2 bacterial liquid with high-temperature sterilized corncob powder passed through a 40-mesh sieve at a volume-to-mass ratio of 2:1 (mL / g); dry at room temperature for 48 hours until the moisture content is ≤25% , to obtain Brevibacterium cold-tolerant ZL-2 compost bacterial agent.

[0048] In a preferred manner, the preparation method of the Brevibacterium cold-resistant ZL-2 bacterial liquid includes the steps: S1, transferring the Brevibacterium cold-resistant ZL-2 bacterial strain stored on the inclined solid medium to the solid medium plate by streaking, Placed in a constant temperature incubator at 10°C for 72 hours for rejuvenation. Take a single colony of Brevibacterium cold-resistant ZL-2 after rejuvenation, inoculate it into beef extrac...

Embodiment 3

[0052] Embodiment 3: Effect evaluation of application of composting low-temperature start-up bacterial agent

[0053] 1. At an ambient temperature of 5-10°C, establish an evaluation experiment for the effect of composting bacterial agents. Fully mix fresh cow dung and straw at a mass ratio of 4:1, adjust the water content to about 60%, and obtain a cow dung composting system. The Brevibacterium cold-resistant ZL-2 composting agent was inoculated at 0.5% by weight in a cow dung composting system with a length of 1m, a width of 0.8m, and a height of 1.2m, and a blank control group without adding the agent was set.

[0054] 2. Record the ambient temperature and the temperature of the pile every day. The temperature of the pile is measured by a piercing thermometer. The measurement position is 30cm below the height of the center of the pile. The temperature of the pile is measured once a day at 9:00 and 17:00 and the average value is taken.

[0055] 3. The experiment did not carr...

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Abstract

The invention discloses a strain of Brevibacterium cold-resistant ZL-2 and a preparation method of a low-temperature start-up agent for composting Brevibacterium cold-resistant ZL-2, comprising the steps of making the number of effective viable bacteria > 1×10 8 Mix the CFU / mL Brevibacterium cold-resistant ZL-2 bacterial liquid with the corncob powder at a volume-to-mass ratio of 2:1 (mL / g); dry it until the moisture content is ≤25%, and then you can get Brevibacterium cold-tolerant ZL-2 compost low temperature Start the inoculant. The Brevibacterium cold-resistant ZL‑2 composting low-temperature starter agent prepared by the invention can be applied to the composting of fresh feces in autumn and winter, and solves the problem of environmental pollution caused by the difficulty in starting fermentation of compost in a cold environment and the accumulation of a large amount of feces. In a low temperature environment of about 5-10°C, the compost heap with the addition of bacterial agents heats up quickly and has a high temperature. It enters a high-temperature period on the 5th to 6th day and lasts for 7-8 days. The highest temperature is 64.5°C, which has reached the level of composting treatment in my country. Harmless Standard (NY / T 1168‑2006).

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the preparation and application of a cold-resistant Brevibacterium and a composting low-temperature starter bacterial agent. Background technique [0002] Low-temperature microorganisms refer to microorganisms that have the ability to grow under low temperature conditions. They are widely distributed in various cold environments and occupy a unique and important position in the ecosystem. Low-temperature microorganisms can generally be divided into psychrophilic and cold-tolerant types. The optimum growth temperature of psychrophilic bacteria is usually ≤15°C, while psychrophilic bacteria refer to microorganisms that can grow well at around 0°C, but the optimum growth temperature is >20°C. The study of low-temperature microorganisms can not only deeply understand the mechanism of microbial cold adaptation, but also its special products such as antifreeze proteins, low-temperature...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C05F15/00C05F17/20C12R1/13
CPCC12N1/20C05F3/00C05F17/00C12R2001/13C12N1/205C05F11/00C05F11/08Y02W30/40
Inventor 李晓宇张喆徐永平陈岩王丽丽侯扶琴陈红莉吴妍妍王思佳
Owner DALIAN UNIV OF TECH
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