WO2017012571A1 - 一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用 - Google Patents

一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用 Download PDF

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WO2017012571A1
WO2017012571A1 PCT/CN2016/090897 CN2016090897W WO2017012571A1 WO 2017012571 A1 WO2017012571 A1 WO 2017012571A1 CN 2016090897 W CN2016090897 W CN 2016090897W WO 2017012571 A1 WO2017012571 A1 WO 2017012571A1
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bacillus coagulans
cctcc
bacillus
laying
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张梁
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Jiangnan University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/07—Bacillus
    • C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus

Definitions

  • the invention relates to the technical field of microorganisms, in particular to the application of a strain of Bacillus coagulans in improving the laying amount of laying hens in the late laying period.
  • microecological preparation plays a role by regulating the growth and reproduction of the normal microbial population and inhibiting the growth and reproduction of the pathogenic bacteria. Due to the outstanding prevention and health effects, micro-ecological preparations can replace antibiotics and chemicals in livestock and poultry farming to achieve true “green, non-polluting”.
  • Microecological preparations are used in laying hens to improve the health of laying hens and promote egg production.
  • CN 103518980B reports a microecological preparation obtained by mixing Lactobacillus plantarum and Bacillus subtilis, which can improve the health of the body after use.
  • CN 101715879B reports a feed additive prepared from an enzyme preparation and Enterococcus faecium which improves the health of the chicken and increases the level of egg production.
  • CN 103202400B reports a microecological preparation compounded by Lactobacillus casei, Lactobacillus plantarum and Bacillus licheniformis, which can improve the performance of broiler chickens and laying hens. These reports prove that microecological preparations are promising in green farming.
  • the applicant provides an application of a strain of Bacillus coagulans to increase the egg production of laying hens.
  • the invention is used in laying hens to significantly improve the laying performance of laying hens in the late laying stage: increasing egg production, increasing egg production rate, and reducing egg-to-egg ratio.
  • the rest is a filler
  • the Bacillus coagulans is Bacillus coagulans ZS-007, the culture collection number is CCTCC No.M 2015273, and the deposit date is May 3, 2015. The deposit is the China Center for Type Culture Collection.
  • the effective viable preparation of the composite microecological preparation is 1.0 ⁇ 10 8 -5.0 ⁇ 10 10 cfu/g, and the amount used is 0.1%-1% of the daily feed intake of the laying hen.
  • 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.
  • the Bacillus coagulans strain was inoculated onto the nutrient agar slant medium, and cultured at 25-37 ° C for 24-48 hr; then, the stalk was transferred to the slant of the nutrient agar eggplant bottle, and cultured at 25-37 ° C for 24-48 hr.
  • composition of nutrient agar slant medium and nutrient agar eggplant bottle are: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, agar 15-20g / L, pH 7.2-7.4.
  • the activated strain was inoculated into a shake flask and shake cultured for 18-30 hr.
  • Shake flask medium formula: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.2-7.4. 250mL shake flask volume is 20mL, 500mL shake flask volume is 50mL.
  • Seed tank medium composition peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.2-7.4.
  • the seed tank has a volume of 10-1000L and a liquid loading of 50-80%.
  • Culture conditions stirring speed 180-250r/min, aeration rate 0.5-1.0vvm, temperature 25-37 ° C, culture time 18-30 hr.
  • the seed liquid is connected to the fermenter with an inoculum of 0.1%-10%.
  • the fermentation medium consists of glucose 25g/L, peptone 15g/L, yeast powder 10g/L, potassium dihydrogen phosphate 1g/L, magnesium sulfate 0.5g. /L, initial pH 7.0.
  • the fermenter has a volume of 1-50 m 3 and a liquid loading of 50-80%.
  • Culture conditions stirring speed 100-200r/min, aeration rate 0.5-0.8vvm, temperature 25-37 ° C, culture time 16-48 hr.
  • the fermentation liquid is taken for microscopic examination, when more than 90% of the cells form spores, the fermentation end point is obtained, and the final fermentation liquid is obtained.
  • the viable cell count and the number of spores of the final fermentation broth were measured by a plate coating method, and the spore rate was calculated.
  • the spore rate is the ratio of the number of spores to the number of viable cells.
  • the medium used was a nutrient agar medium: peptone 10 g/L, beef extract 3 g/L, sodium chloride 5 g/L, agar 15-20 g/L, pH 7.2-7.4.
  • the mixture was determined by gradient dilution; when the number of spores was measured, the final fermentation broth was heat-treated at 80 ° C for 10 min, and the mixture was diluted by a gradient.
  • the number of viable cells in the final fermentation broth was 1.5 ⁇ 10 10 cfu/mL or more, the number of spores was 1.4 ⁇ 10 10 cfu/mL or more, and the spore rate was 90.8% or more.
  • the dry bacterial powder is obtained by a spray drying process.
  • the operating conditions are: inlet air temperature 210 ° C, outlet air temperature 100 ° C.
  • the number of viable bacteria in the dry powder is 8.0 ⁇ 10 11 cfu/g or more.
  • the beneficial technical effect of the invention is that: in the laying hen culture, the laying performance of the laying hen in the late laying period can be significantly improved: the egg production is increased by more than 11%, the egg production rate is increased by 9.2%, and the egg-to-egg ratio is reduced by 10%. %the above.
  • 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.
  • the obtained strain was inoculated on a nutrient agar slant medium, cultured at 30 ° C for 24 hr, and then streaked to the slant of the nutrient agar eggplant bottle, and cultured at 30 ° C for 24 hr.
  • the composition of nutrient agar slant medium and nutrient agar eggplant bottle are: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, agar 15-20g / L, pH 7.2-7.4.
  • Shake flask culture The activated strain was inoculated into a shake flask and shake cultured for 20 hr.
  • Shake flask medium formula: peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.2-7.4, shake flask volume of 50mL / 500mL triangle bottle.
  • Culture conditions shake flask speed 200r/min, temperature 30 °C.
  • Seed tank expansion Collect 20 bottles of 500 mL shake flask seed solution for seed tank inoculation.
  • Seed tank medium composition peptone 10g / L, beef extract 3g / L, sodium chloride 5g / L, pH 7.2-7.4.
  • the seed tank has a volume of 500L and a liquid loading of 75%.
  • Culture conditions stirring speed 200 r / min, aeration rate 0.8 vvm, temperature 30 ° C, incubation time 20 hr.
  • Fermentor Fermentation The liquid volume of the 30m 3 fermenter is 70%.
  • the composition of the medium was glucose 25 g/L, peptone 15 g/L, yeast powder 10 g/L, potassium dihydrogen phosphate 1 g/L, magnesium sulfate 0.5 g/L, initial pH 7.0.
  • the cultured seeds are connected to a fermentor for fermentation.
  • Culture conditions stirring speed 100 r / min, aeration amount 0.6 vvm, temperature 35 ° C, culture time 40-45 hr.
  • the number of viable cells was 1.99 ⁇ 10 10 cfu/g
  • the number of spores was 1.92 ⁇ 10 10 cfu/g
  • the spore rate was 96.5%.
  • the wet bacterial sludge was collected by centrifugation, and a dry powder was obtained by a spray drying process.
  • the operating conditions are: inlet air temperature 210 ° C, outlet air temperature 100 ° C.
  • the number of viable bacteria in the dried powder was 8.32 ⁇ 10 11 cfu / g.
  • Bacillus coagulans prepared in Example 1 were combined with Bacillus subtilis CCTCC No. M 2011443 and Bacillus licheniformis CCTCC No. M 2012458 to prepare a composite microecological preparation for improving laying capacity of laying hens; Bacillus subtilis CCTCC No. M 2011443, Bacillus licheniformis CCTCC No.M 2012458 single powder was purchased from Taizhou Green Shengyuan Biotechnology Co., Ltd., its product specifications are 5.0 ⁇ 10 10 cfu / g.
  • Strain Per gram of product contains viable cell count (cfu) Bacillus coagulans CCTCC No.M 2015273 0.4 ⁇ 10 8 -2.0 ⁇ 10 10 Bacillus subtilis CCTCC No.M 2011443 0.4 ⁇ 10 8 -2.0 ⁇ 10 10 Bacillus licheniformis CCTCC No.M 2012458 0.2 ⁇ 10 8 -1.0 ⁇ 10 10
  • the Bacillus coagulans prepared in Example 1 were combined with Bacillus subtilis CCTCC No. M 2011443 and Bacillus licheniformis CCTCC No. M 2012458 to prepare a composite microecological preparation for improving the laying amount of laying hens, and the content of each strain was as shown in Table 3.
  • the remaining material per gram of product is (filler (glucose, stone powder, etc.)
  • Strain Per gram of product contains viable cell count (cfu) Bacillus coagulans CCTCC No.M 2015273 0.4 ⁇ 10 8 Bacillus subtilis CCTCC No.M 2011443 0.4 ⁇ 10 8 Bacillus licheniformis CCTCC No.M 2012458 0.2 ⁇ 10 8
  • the Bacillus coagulans prepared in Example 1 were compounded with Bacillus subtilis CCTCC No. M 2011443 and Bacillus licheniformis CCTCC No. M 2012458 to obtain the egg production of the hens.
  • the composite microecological preparations, the contents of each strain are shown in Table 4.
  • the remaining material per gram of product is (filler (glucose, stone powder, etc.)
  • Strain Per gram of product contains viable cell count (cfu) Bacillus coagulans CCTCC No.M 2015273 2.0 ⁇ 10 10 Bacillus subtilis CCTCC No.M 2011443 2.0 ⁇ 10 10 Bacillus licheniformis CCTCC No.M 2012458 1.0 ⁇ 10 10
  • the Bacillus coagulans prepared in Example 1 were combined with Bacillus subtilis CCTCC No. M 2011443 and Bacillus licheniformis CCTCC No. M 2012458 to prepare a composite microecological preparation for improving the laying amount of laying hens, and the content of each strain was As shown in Table 5.
  • the remaining material per gram of product is (filler (glucose, stone powder, etc.)
  • Strain Per gram of product contains viable cell count (cfu) Bacillus coagulans CCTCC No.M 2015273 1.0 ⁇ 10 9 Bacillus subtilis CCTCC No.M 2011443 1.0 ⁇ 10 9 Bacillus licheniformis CCTCC No.M 2012458 1.0 ⁇ 10 9
  • the composite microecological preparation prepared in Example 3 was used for feeding the laying hens.
  • the test site is located in an ecological pasture laying hen site in Qidong, Nantong, Jiangsu.
  • the test chickens were kept in large sheds, and the eggs were laid for 500 days.
  • the peak rate of high egg production was 90%. 46 per shed, free to eat and drink, according to the routine feeding management.
  • each group used the same feeding management method.
  • the pre-test period was 7 days, and the trial period was 60 days.
  • the hen ecological farming method the four areas with the same area and ecological environment were divided into sections and the same area.
  • the laying rate of laying hens increased by 9.2% in the late egg laying period, and the total egg production increased by more than 11%.
  • the egg ratio has dropped by more than 10%.

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Abstract

提供了一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用，所述凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备得到提升蛋鸡产蛋量的复合微生态制剂；所述凝结芽孢杆菌为凝结芽孢杆菌(Bacillus coagulans)ZS-007，菌种保藏编号为CCTCC No.M 2015273，保藏日期为2015年5月3日，保藏地点为中国典型培养物保藏中心。

Description

一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用 技术领域
本发明涉及微生物技术领域，尤其是涉及一株凝结芽孢杆菌在提高蛋鸡在产蛋后期产蛋量上的应用。
背景技术
随着生活水平日益提高，人们越来越关注抗生素、化学药品残留所引发的食品安全和生活健康问题，对无药物残留的绿色畜禽产品的需求越来越多。微生态制剂通过调节促进正常微生物群生长繁殖、抑制致病菌生长繁殖而发挥作用。因突出的预防和保健效果，微生态制剂在畜禽养殖中可替代抗生素、化学药品，实现真正的“绿色、无污染”。
鸡蛋早已成为人们日常生活的必需品。微生态制剂用在蛋鸡养殖中，可以提高蛋鸡健康状态、促进产蛋量。CN 103518980B报道了一种由植物乳杆菌和枯草芽孢杆菌混合所得到的微生态制剂，使用后，可以提高机体健康程度。CN 101715879B报道了一种由酶制剂和屎肠球菌制备的饲料添加剂，能改善鸡只健康状态，提高产蛋水平。CN 103202400B报道了由干酪乳杆菌、植物乳杆菌、地衣芽孢杆菌复配的微生态制剂，能改善肉鸡、蛋种鸡生产性能。这些报道，证明微生态制剂在绿色养殖中大有可为。
一般来说，鸡群在开产300天后淘汰低产个体，开产500天后产蛋显著下降，必要时需要更新鸡群。如何提升蛋鸡在产蛋后期的产蛋性能，最大化利用饲料，相关研究很少。
发明内容
针对现有技术存在的上述问题，本申请人提供了一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用。本发明用于蛋鸡养殖中，可显著提升蛋鸡在产蛋后期的产蛋性能：提高产蛋量、增加产蛋率，降低料蛋比。
本发明的技术方案如下：
一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用，所述凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备 得到提升蛋鸡产蛋量的复合微生态制剂；所述三种菌种的用量分别为：
Figure PCTCN2016090897-appb-000001
其余为填充物；
所述凝结芽孢杆菌为凝结芽孢杆菌(Bacillus coagulans)ZS-007，菌种保藏编号为CCTCC No.M 2015273，保藏日期为2015年5月3日，保藏地点为中国典型培养物保藏中心。
所述复合微生态制剂有效活菌数为1.0×108-5.0×1010cfu/g，使用量为蛋鸡日采食量的0.1％-1％。
1、凝结芽孢杆菌的筛选
从江苏无锡马山某奶牛养殖场附近的土壤样品称取2g，加入98ml无菌生理盐水，80℃热处理10min后，在37℃摇床上120rpm条件下振荡20min，吸取上清液1ml加入LB培养基中稀释涂布进行菌株分离。通过对菌株进行形态学观察、生理生化实验、16S rDNA分析等手段有效而准确的确定菌种的属种。
最终得到编号为ZS-007的菌种，鉴定为凝结芽孢杆菌(Bacillus coagulans)。
ZS-007于2015年5月3日保藏于中国典型培养物保藏中心，保藏编号为CCTCC M 2015273。
2、凝结芽孢杆菌高密度发酵
(1)菌种活化
将凝结芽孢杆菌菌种接种至营养琼脂斜面培养基上，25-37℃下培养24-48hr；然后划线转接于营养琼脂茄子瓶斜面，25-37℃下培养24-48hr。
营养琼脂斜面培养基及营养琼脂茄子瓶的组成均为：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，琼脂15-20g/L，pH7.2-7.4。
(2)摇瓶培养
将活化好的菌种接种至摇瓶中，振荡培养18-30hr。
摇瓶培养基配方：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，pH7.2-7.4。250mL摇瓶装液量为20mL，500mL摇瓶装液量为50mL。
培养条件：摇瓶转速200r/min，温度25-37℃。
(3)种子罐扩培
扩培菌种以用于后期发酵。种子罐培养基组成：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，pH7.2-7.4。种子罐体积10-1000L，装液量50-80％。
培养条件：搅拌转速180-250r/min，通气量0.5-1.0vvm，温度25-37℃，培养时间18-30hr。
(4)发酵罐发酵
以0.1％-10％的接种量将种子液接入发酵罐，发酵培养基组成：葡萄糖25g/L，蛋白胨15g/L，酵母粉10g/L，磷酸二氢钾1g/L，硫酸镁0.5g/L，初始pH7.0。发酵罐体积1-50m3，装液量50-80％。
培养条件：搅拌转速100-200r/min，通气量0.5-0.8vvm，温度25-37℃，培养时间16-48hr。
取发酵液镜检时，当90％以上的菌体都形成芽孢时，即到发酵终点，得到最终发酵液。
采用平板涂布法测定最终发酵液活菌数和芽孢数，并计算芽孢率。其中，芽孢率为芽孢数与活菌数的比值。所用培养基为营养琼脂培养基：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，琼脂15-20g/L，pH7.2-7.4。测定最终发酵液活菌数时，梯度稀释后测定；测定芽孢数时，取最终发酵液于80℃热处理10min，梯度稀释后测定。最终发酵液中活菌数1.5×1010cfu/mL以上、芽孢数1.4×1010cfu/mL以上、芽孢率90.8％以上。
3、凝结芽孢杆菌干菌粉制备
最终发酵液离心后收集湿菌泥，采用喷雾干燥工艺得到干菌粉。操作条件为：进风温度210℃，出风温度100℃。干菌粉中活菌数8.0×1011cfu/g以上。
本发明有益的技术效果在于：用于蛋鸡养殖中，可显著提升蛋鸡在产蛋后期的产蛋性能：产蛋量增加11％以上、产蛋率增加9.2％以上，料蛋比降低10％以上。
具体实施方式
下面结合实施例，对本发明进行具体描述。
实施例1
1、凝结芽孢杆菌的筛选
从江苏无锡马山某奶牛养殖场附近的土壤样品称取2g，加入98ml无菌生理 盐水，80℃热处理10min后，在37℃摇床上120rpm条件下振荡20min，吸取上清液1ml加入LB培养基中稀释涂布进行菌株分离。通过对菌株进行形态学观察、生理生化实验、16S rDNA分析等手段有效而准确的确定菌种的属种。
最终得到编号为ZS-007的菌种，鉴定为凝结芽孢杆菌(Bacillus coagulans)。
ZS-007于2015年5月3日保藏于中国典型培养物保藏中心，保藏编号为CCTCC M 2015273。
2、30m3发酵罐制备
菌种活化：将得到的菌种接种至营养琼脂斜面培养基上，30℃下培养24hr后划线转接于营养琼脂茄子瓶斜面，30℃下培养24hr。营养琼脂斜面培养基及营养琼脂茄子瓶的组成均为：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，琼脂15-20g/L，pH7.2-7.4。
摇瓶培养：将活化好的菌种接种至摇瓶中，振荡培养20hr。摇瓶培养基配方：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，pH7.2-7.4，摇瓶装液量50mL/500mL三角瓶。培养条件：摇瓶转速200r/min，温度30℃。
种子罐扩培：收集20瓶500mL摇瓶种子液用于种子罐接种。种子罐培养基组成：蛋白胨10g/L，牛肉膏3g/L，氯化钠5g/L，pH7.2-7.4。种子罐体积500L，装液量75％。培养条件：搅拌转速200r/min，通气量0.8vvm，温度30℃，培养时间20hr。
发酵罐发酵：30m3发酵罐装液量70％。培养基组成：葡萄糖25g/L，蛋白胨15g/L，酵母粉10g/L，磷酸二氢钾1g/L，硫酸镁0.5g/L，初始pH7.0。将培养好的种子接入发酵罐进行发酵。培养条件：搅拌转速100r/min，通气量0.6vvm，温度35℃，培养时间40-45hr。
发酵液中，活菌数1.99×1010cfu/g，芽孢数1.92×1010cfu/g，芽孢率96.5％。
3、干菌粉制备
离心收集湿菌泥，采用喷雾干燥工艺得到干菌粉。操作条件为：进风温度210℃，出风温度100℃。干菌粉中活菌数8.32×1011cfu/g。
由实施例1制备得到的凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备得到提升蛋鸡产蛋量的复合微生态制剂；枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC  No.M 2012458的单菌粉采购自泰州绿圣元生物科技有限公司，其产品规格均为5.0×1010cfu/g。
每500g复合微生态制剂中，各物质含量如表1所示：
表1
名称 重量
凝结芽孢杆菌CCTCC No.M 2015273 0.4g-200g
枯草芽孢杆菌CCTCC No.M 2011443 0.4g-200g
地衣芽孢杆菌CCTCC No.M 2012458 0.2g-100g
填充物(葡萄糖、石粉等) 499g-0g
折算后，复合微生态制剂中各微生物含量如表2所示：
表2
菌种 每克产品含有菌种活菌数(cfu)
凝结芽孢杆菌CCTCC No.M 2015273 0.4×108-2.0×1010
枯草芽孢杆菌CCTCC No.M 2011443 0.4×108-2.0×1010
地衣芽孢杆菌CCTCC No.M 2012458 0.2×108-1.0×1010
实施例2
由实施例1制备得到的凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备得到提升蛋鸡产蛋量的复合微生态制剂，每种菌种的含量如表3所示。每克产品中剩余的为填充物(葡萄糖、石粉等)
表3
菌种 每克产品含有菌种活菌数(cfu)
凝结芽孢杆菌CCTCC No.M 2015273 0.4×108
枯草芽孢杆菌CCTCC No.M 2011443 0.4×108
地衣芽孢杆菌CCTCC No.M 2012458 0.2×108
实施例3
由实施例1制备得到的凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备得到提升蛋鸡产蛋量的 复合微生态制剂，每种菌种的含量如表4所示。每克产品中剩余的为填充物(葡萄糖、石粉等)
表4
菌种 每克产品含有菌种活菌数(cfu)
凝结芽孢杆菌CCTCC No.M 2015273 2.0×1010
枯草芽孢杆菌CCTCC No.M 2011443 2.0×1010
地衣芽孢杆菌CCTCC No.M 2012458 1.0×1010
实施例4
由实施例1制备得到的凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M 2011443、地衣芽孢杆菌CCTCC No.M 2012458复配制备得到提升蛋鸡产蛋量的复合微生态制剂，每种菌种的含量如表5所示。每克产品中剩余的为填充物(葡萄糖、石粉等)
表5
菌种 每克产品含有菌种活菌数(cfu)
凝结芽孢杆菌CCTCC No.M 2015273 1.0×109
枯草芽孢杆菌CCTCC No.M 2011443 1.0×109
地衣芽孢杆菌CCTCC No.M 2012458 1.0×109
测试例：
用实施例3制备得到的复合微生态制剂进行蛋鸡喂养。试验地点位于江苏南通启东某生态牧场蛋鸡场。试验鸡为大棚散养，产蛋500天，产蛋率较高峰期90％的水平已显著下降。每棚46只，自由采食和饮水，按常规进行饲养管理。试验期间各组采用相同的饲养管理方式，预试期7d，正试期为60d，按母鸡生态养殖方法在圈一块地并划分围成面积、生态环境相同的四片区，每片区均设相同采食、采水槽、产蛋箱、鸡舍，每组在各片区轮换饲养，每组在每片区饲养20d，各片区轮换时彻底消毒清理，以消除位置效应的影响，试验期间早晚补饲日粮，自由采食与饮水。正式试验期从2015年1月20日至2015年3月21日。试验结果如表6所示：
表6
Figure PCTCN2016090897-appb-000002
从上表可以看到，使用包括凝结芽孢杆菌CCTCC No.M 2015273在内的复合微生态制剂，蛋鸡在产蛋后期的产蛋率增加9.2％以上、产蛋总量增加11％以上、料蛋比下降10％以上。

Claims (2)

  1. 一株凝结芽孢杆菌在提升蛋鸡产蛋量上的应用，其特征在于所述凝结芽孢杆菌与枯草芽孢杆菌CCTCC No.M2011443、地衣芽孢杆菌CCTCC No.M2012458复配制备得到提升蛋鸡产蛋量的复合微生态制剂；所述三种菌种的用量分别为：
    Figure PCTCN2016090897-appb-100001
    所述凝结芽孢杆菌为凝结芽孢杆菌(Bacillus coagulans)ZS-007，菌种保藏编号为CCTCC No.M2015273，保藏日期为2015年5月3日，保藏地点为中国典型培养物保藏中心。
  2. 根据权利要求1所述的凝结芽孢杆菌在提升蛋鸡产蛋量上的应用，其特征在于所述复合微生态制剂有效活菌数为1.0×108-5.0×1010cfu/g，使用量为蛋鸡日采食量的0.1％-1％。
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