WO2017012573A1

WO2017012573A1 - Bacillus pumilus strain and microbial ecological combined agent thereof for weaned piglet

Info

Publication number: WO2017012573A1
Application number: PCT/CN2016/090899
Authority: WO
Inventors: 张梁
Original assignee: 江南大学
Priority date: 2015-07-22
Filing date: 2016-07-21
Publication date: 2017-01-26
Also published as: AU2016297405B2; AU2016297405A1; CN104974967B; CN104974967A

Abstract

Provided is a Bacillus pumilus strain (serial number: ZS008) deposited in the China Center for Type Culture Collection (CCTCC) on July 13, 2015 and having a deposition number of CCTCC No. M2015451. Also provided is a microbial ecological Bacillus combined agent for a weaned piglet, comprising the following Bacillus species: a Bacillus pumilus (CCTCC No. M2015451), a Bacillus coagulans (CCTCC No. M2015273), a Bacillus subtilis (CCTCC No. M2011443) and a Bacillus licheniformis (CCTCC No. M2012458).

Description

Composite microecological preparation for a strain of Bacillus pumilus and its nursery piglets

Technical field

The invention relates to the field of microbial technology, in particular to a strain of Bacillus brevis and its use in the conservation of piglets.

Background technique

Conservation piglets, referred to as weaned piglets, pigs at this stage are quite fragile, maternal antibodies disappear, new immune system has not yet been established, coupled with rapid growth, stress factors, etc., it is prone to various diseases and even large The area is dead. Although antibiotics and chemicals can solve the problems in a short period of time, the resulting drug resistance and drug residue problems may affect the growth of piglets in the later period. How to improve the health status, survival rate, uniformity and full effect of growth of piglets is a common concern of every farmer.

The microecological preparation has the functions of regulating intestinal microflora balance, proliferating beneficial bacteria, inhibiting harmful bacteria, lowering intestinal pH value, increasing digestive enzyme activity, nutrient digestibility and immunity, and microecological preparation in piglet breeding. The application reports are increasing. CN 101703136B reports the application of the Lactobacillus plantarum preparation in the prevention and treatment of piglet diarrhea, the efficiency of piglet diarrhea is more than 90%, and the cure rate is over 80%. CN 103255080B reports the use of Bacillus coagulans HM-09 in the preparation of feed for improving the performance of piglets and diarrhea, which can reduce the incidence of diarrhea in weaned piglets, thereby reducing the incidence of intestinal diseases and improving feed digestion. Rate to improve feed utilization of piglets. CN 103205373B reported a Clostridium butyricum strain for suckling pigs, which can antagonize animal pathogens in the intestinal tract of suckling pigs, maintain and regulate micro-ecological balance, enhance immune function, produce starch plum and vitamins, and promote animal feed. Digestive absorption and nutritional effects.

In general, microecological preparations have a slow onset of action, and they need to be used consistently in livestock and poultry farming to receive good results. Therefore, the development of a microecological preparation that can not only long-term health care, but also quickly solve the problems of piglet diarrhea, diarrhea, etc. is particularly necessary for the stable production of nursery piglets.

Summary of the invention

In view of the above problems existing in the prior art, the applicant provides a composite microecological preparation for Bacillus pumilus and its nursery piglets. The invention can quickly solve the problems of piglet diarrhea, squatting, etc. Improve the health of nursery pigs during long-term use.

The technical solution of the present invention is as follows:

A strain of Bacillus pumilus ZS008 was deposited with the China Center for Type Culture Collection under the accession number CCTCC No. M2015451 and the date of deposit was July 13, 2015.

A compound Bacillus subtilis preparation for nursery pigs is prepared by combining the following strains into a composite fungus: Bacillus pumilus CCTCC No. M2015451, Bacillus coagulans CCTCC No. M2015273, Bacillus subtilis CCTCC No. M 2011443, lichens spore Bacillus CCTCC No. M 2012458.

The effective viable cell count of the composite Bacillus subtilis preparation is 1.4×10 ⁸ -7.0×10 ¹⁰ cfu/g, and the composition of each strain is as follows:

The Bacillus coagulans is Bacillus coagulans ZS007 and is deposited with the China Center for Type Culture Collection under the accession number CCTCC NO: M 2015273.

The application of the composite Bacillus subtilis preparation for the nursery piglets is to improve the performance of the nursery piglets. The amount used is 1.0 to 5.0 g/head·day.

The single bacterial powder of Bacillus subtilis CCTCC No.M 2011443 and Bacillus licheniformis CCTCC No.M 2012458 was purchased from Taizhou Green Shengyuan Biotechnology Co., Ltd., and its product specifications were all 5.0×10 ¹⁰ cfu/g.

The beneficial technical effects of the present invention are:

(1) It can quickly solve problems such as diarrhea and squatting of piglets;

(2) After long-term use, the health level of the nursery pigs can be improved, so as to improve the production capacity of the nursery pigs.

detailed description

1. Screening of Bacillus brevis

2 g of soil samples from Fangding Ding pig farm in Wuxi, Jiangsu Province were weighed, 98 ml of sterile physiological saline was added, heat treated at 80 ° C for 10 min, shaken at 120 rpm on a 37 ° C shaker for 20 min, and 1 ml of the supernatant was aspirated into LB medium. The strain was separated by dilution coating. The species of the strain were effectively and accurately determined by means of morphological observation, physiological and biochemical experiments, and 16S rDNA analysis.

The strain numbered ZS008 was finally obtained and identified as Bacillus pumilus. The strain is deposited in the China Center for Type Culture Collection under the accession number CCTCC No.M2015451 and the deposit date is July 13, 2015.

2. Screening of Bacillus coagulans

Weigh 2g from the soil sample near a dairy farm in Mashan, Wuxi, Jiangsu, add 98ml of sterile physiological saline, heat treatment at 80 °C for 10min, shake at 120rpm on a 37 °C shaker for 20min, and absorb 1ml of supernatant to add LB medium. The strain was isolated by medium dilution coating. The species of the strain were effectively and accurately determined by means of morphological observation, physiological and biochemical experiments, and 16S rDNA analysis.

The strain numbered ZS-007 was finally obtained and identified as Bacillus coagulans.

ZS-007 was deposited with the China Type Culture Collection on May 3, 2015 under the accession number CCTCC M 2015273.

3. Preparation of composite microecological preparations

The composite microecological preparation is compounded by Bacillus pumilus CCTCC No. M2015451, Bacillus coagulans CCTCC No. M 2015273, Bacillus subtilis CCTCC No. M 2011443, and Bacillus licheniformis CCTCC No. M 2012458.

The single bacterial powder of Bacillus subtilis CCTCC No. M 2011443 and Bacillus licheniformis CCTCC No. M 2012458 was purchased from Taizhou Green Shengyuan Biotechnology Co., Ltd., and its product specifications were all 5.0×10 ¹⁰ cfu/g.

The microecological preparation products prepared in Examples 1 to 3, the contents of each microorganism are as follows:

Table 1

After the single bacteria powder is uniformly mixed according to the predetermined ratio, anhydrous glucose is added as a carrier to supplement 100% of the total weight (Bacillus bacillus is used as an active ingredient, and the total weight of the product is satisfied to reach the final viable cell count of 4.0×10 ⁸ cfu. /g, mix well and set aside.

The processing should ensure the cleanliness of the operating environment and avoid contamination. Control the ambient humidity below 70%, the moisture content of each raw material is less than 13%, the product is stored at room temperature for 24 months after being made, the shelf life at -18 °C can reach more than 60 months, and the number of viable bacteria in the shelf life is stable.

4. Application effect of compound probiotics on nursery piglets

Choose from the same batch of self-property and the same batch of Du × Chang × Da Sanyuan hybrid weaned piglets 90. According to the principle of parity, body weight, male or female ratio, the piglets were divided into three groups: A, B and C. Group A was the control group, B test group 1 and C group were the test group 2, each group Set 2 repetitions, 15 repetitions each, randomize and divide the number; the basic diet of each group is the same (Zhengda full price), and the nutritional level is consistent. Group A did not add any enzyme preparation, and the group B basic diet was added with a composite microecological preparation containing Bacillus pumilus (Example 1), Group C was the same as Group B, but did not contain Bacillus pumilus; the amount of microecological preparation was The amount used is 1.0 g / head · day. The piglets were 21 days old, weaned at an average weight of 6.5 kg, pre-tested for 7 days, and fed for 30 days. A breeding cooperative in Nanjing.

The test results are shown in Tables 2 to 4.

Table 2 Weight gain of test pigs and average daily gain during test period

Table 3 Feed remuneration of each group fed with enzyme-probiotics

Table 4 Frequency of piglet diarrhea in each group fed with enzyme-probiotics

After the piglets have diarrhea, they are treated with microecological preparations at a dose of 5g per pig. After the product was stirred with 10 mL of cold boiled water, the syringe was drawn into the syringe and filled with a syringe along the mouth of the piglet. When the microecological preparation product contains Bacillus brevis, the piglets improved after 4-6 hrs, and the diarrhea was terminated within 24 hr, and the appetite was restored. When the microecological preparation product does not contain Bacillus brevis, the piglets turn better after 12 hours; the treatment is consolidated once, and the diarrhea is terminated in about 2 days to restore appetite.

Claims

A strain of Bacillus pumilus ZS008 is deposited with the China Center for Type Culture Collection under the accession number CCTCC NO.M2015451 and the date of deposit is July 13, 2015.
The invention relates to a compound Bacillus subtilis preparation for protecting piglets, which is characterized in that the combination of the following strains is compounded into a composite fungus: Bacillus pumilus CTCCC NO.M2015451, Bacillus coagulans CTCCC NO.M2015273, Bacillus subtilis CCTCC NO.M2011443, Bacillus licheniformis CCTCC NO. M2012458.
The composite Bacillus subtilis preparation for nursery piglets according to claim 2, wherein the effective viable cell count of the Bacillus subtilis preparation is 1.4×10 8 -7.0×10 10 cfu/g, each strain The composition is as follows:
The composite Bacillus subtilis preparation for nursery piglets according to claim 2, wherein the Bacillus coagulans is Bacillus coagulans ZS007, and is deposited in the China Center for Type Culture Collection under the accession number CCTCC NO: M2015273.
The use of the composite Bacillus subtilis formulation for a nursery pig according to claim 2, characterized in that the performance of the nursery pig is improved.
The use of the composite Bacillus subtilis preparation for nursery piglets according to claim 5, wherein the Bacillus subtilis preparation is used in an amount of 1.0 to 5.0 g per head per day.