CN105624071A - Lactobacillus salivarius XJP2 and application thereof - Google Patents
Lactobacillus salivarius XJP2 and application thereof Download PDFInfo
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- CN105624071A CN105624071A CN201610152385.2A CN201610152385A CN105624071A CN 105624071 A CN105624071 A CN 105624071A CN 201610152385 A CN201610152385 A CN 201610152385A CN 105624071 A CN105624071 A CN 105624071A
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Abstract
The invention provides lactobacillus salivarius XJP2 and application thereof. The lactobacillus salivarius XJP2 is separated from the cecum mucosa of a healthy broiler chicken and preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No.11386. The lactobacillus salivarius XJP2 has the better tolerance to acids and cholate and can be colonized in the animal intestinal tract. The lactobacillus salivarius XJP2 is high in in-vitro culture reproduction speed and L-lactic acid yield, has the functions of preventing bacterial diarrhea of the broiler chickens, improving the intestinal flora and the like and can be applied to development of microecological preparations.
Description
Technical field
The present invention relates generally to microbial technology field, is specifically related to strain lactobacillus salivarius XJP2 and an application thereof.
Background technology
Under normal circumstances, intestinal microflora and animal body remain equilibrium relationship, maintain the state of health of body: acted on the one hand the pathogenic bacteria contention mucous membrane attachment site entering body by mycoderm barrier or competition exclusion, make the pathogenic bacteria cannot field planting, enhancing body immunizing power, reduces sickness rate; Bacterium group participates in the substance metabolism of body on the other hand, nutrition conversion and synthesis etc., promotes animal health growth. Milk-acid bacteria is one of profitable strain in animal body, is the normal Macroflora colonization of animal intestinal, has biological antagonist, improves animal intestinal ecotope, improves the multiple effects such as animal immunizing power, has been widely used in animal cultivation.
At present, the research and apply of milk-acid bacteria is more, and market associated product category is various, but common problem to be milk-acid bacteria poor to the tolerance of acid, cholate, intestinal mucosa can be colonizated in, survive for a long time in enteron aisle and play a role. Milk-acid bacteria only adheres at alimentary canal mucous membrane after taking in, field planting, just suppress pathogenic bacteria breeding by competitive exclusion, produce meta-bolites or physiologically active substance kill harmful pathogenic micro-organism, and then reach recovery host's resistibility, repair intestinal microflora barrier, cure the effect of intestinal tract disease. So screening can on animal digestion mucous membrane the bacterial classification of field planting most important.
Summary of the invention
The present invention provides strain lactobacillus salivarius XJP2 and an application thereof, and described lactobacillus salivarius XJP2 has preferably acid tolerance and Bile salt resistance, can in enteron aisle Colonization, also can prevent broiler chicken bacterial diarrhea simultaneously, there is the effect of regulating intestinal canal bacterium group.
For achieving the above object, the present invention is achieved by following technical proposals:
The present invention provides a strain lactobacillus salivarius XJP2, and its called after lactobacillus salivarius Lactobacillussalivarius that classifies, its deposit number is CGMCCNo.11386.
Further: described lactobacillus salivarius XJP2 is Gram-positive, and bacterium colony is circular, and neat in edge, oyster white, bacterium colony is smooth.
Further: after described lactobacillus salivarius XJP2 cultivates 20h, Pfansteihl output is greater than 18.2g/L.
Further: acid, cholate are had good tolerance by described lactobacillus salivarius XJP2.
Present invention also offers described lactobacillus salivarius XJP2 for the preparation of the application in the micro-ecological additive of broiler breeding.
Further: described lactobacillus salivarius XJP2 can significantly improve lactic acid bacterium number in caecum.
Further: described lactobacillus salivarius XJP2 can prevent broiler chicken bacterial diarrhea.
Compared with prior art, the present invention has the following advantages and technique effect: the present invention screens a strain lactobacillus salivarius XJP2 from healthy broiler chicken caecum mucous membrane, can at animal intestinal Colonization; Acid and cholate had good tolerance; And the breeding of lactobacillus salivarius XJP2 vitro culture is fast, can be used for the exploitation of probiotics, is applied in broiler breeding, there is prevention bacterial diarrhea, regulate the function of microbial population of animal intestinal tract.
Accompanying drawing explanation
Fig. 1 is the colonial morphology of described lactobacillus salivarius XJP2 on MRS flat board;
Fig. 2 is described lactobacillus salivarius XJP2 growth curve;
Fig. 3 is described caecum Digesta samples DGGE experimental result.
Embodiment
In conjunction with following specific examples, the technical scheme of the present invention is described in further detail.
The screening of embodiment 1, lactobacillus salivarius
1, the separation screening of lactobacillus salivarius XJP2
Taking healthy broiler chicken as sample, scraping caecum mucous membrane in sterilized water with sterilized slide glass, vibration is mixed even carries out gradient dilution, and MRS substratum pours into cultivates screening " molten calcium circle " significantly bacterium colony, repeatedly obtains single bacterium colony after purifying, called after XJP2, preserves. Described MRS substratum is the collective media cultivating milk-acid bacteria, and its formula is: peptone 10g/L, extractum carnis 10g/L, yeast powder 5g/L, glucose 20g/L, tween 80 1ml/L, dipotassium hydrogen phosphate 2g/L, sodium acetate 5g/L, Triammonium citrate 2g/L, magnesium sulfate 0.58g/L, manganous sulfate 0.25g/L, calcium carbonate 5g/L, pH6.2.
Described strain X JP2 MRS flat board colonial morphology as shown in Figure 1, its biological characteristics: the inspection of Gram-positive, mirror is for rod-short; Bacterium colony is circular, smooth, and neat in edge is more moistening, oyster white. Physiological and biochemical index display strain X JP2 can not reduce nitrate, not liquefy gelatin, does not produce indoles and hydrogen sulfide, and catalase test is negative.
2, the 16SrRNA sequencing of lactobacillus salivarius XJP2
Taking the DNA of described strain X JP2 as template, the general primer of 16SrRNA increases, and amplified fragments carries out sequencing; The 16srRNA sequence homology degree that Blast comparison shows Lactobacillussalivarius in this bacterial strain sequence and GenBank gene pool is the highest, and homology is 99%. By DNAMAN6.0, the 16SrRNA sequence of lactobacillus existing in Genbank being carried out genetic evolution analysis, result shows, and strain X JP2 and Lactobacillussalivarius homology are the highest, judge that this strain X JP2 is as lactobacillus salivarius.
The strain X JP2 screened is carried out culture presevation, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC); Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; Preservation date: on September 16th, 2015; The deposit number of lactobacillus salivarius Lactobacillussalivarius is CGMCCNo.11386.
The growth measurement of embodiment 2, lactobacillus salivarius XJP2
By the XJP2 bacterium liquid of activation with 1% (about 107Cfu/mL) inoculum size is inoculated in described MRS liquid nutrient medium, 37 DEG C, and 160rpm cultivates 24h, every 3h sampling, measures light absorption value under 600nm, draws growth curve chart. As shown in Figure 2, experimental result shows, XJP2 cultivates 3h and namely enters logarithmic phase, and 12h enters stationary phase. When cultivating 20h, Pfansteihl output is 18.2g/L.
The tolerance of acid, cholate is measured by embodiment 3, lactobacillus salivarius XJP2
By the XJP2 bacterium liquid of activation with 1% (about 107Cfu/mL) inoculum size is inoculated in the MRS substratum that pH is 2.5,3.5,4.5 respectively, 37 DEG C of quiescent culture. In 0h, 2h, 4h sampling, MRS pours into counting. Process bacterial strain by same method, bacterium liquid is inoculated in the MRS substratum that pig gallbladder salinity is 0.1%, 0.2%, 0.3%, 37 DEG C of quiescent culture. In 0h, 2h, 4h sampling, MRS pours into counting.
From table 1, table 2 result it may be seen that acid, cholate are had good tolerance by strain X JP2, after cultivating 2h in the MRS substratum of pH2.5 or 0.3% cholate, Viable detection is all more than 70%, has good resistance.
Table 1 strain X JP2 is to acid tolerability results (cfu/mL)
Table 2 strain X JP2 is to Bile salt resistance result (cfu/mL)
Embodiment 4, lactobacillus salivarius XJP2 prevent the test of broiler chicken bacterial diarrhea
Select broiler chicken 100 age on the 1st, it is divided into 2 groups at random, often organize 100. Test group adds lactobacillus salivarius bacterium liquid in drinking water, and addition is 106Cfu/L, control group does not add bacterium liquid, all feeds by normal procedure for two groups, observes, adds up diarrhea rate, surviving rate. Between trial period, test group surviving rate is obviously higher than control group, and rate of death caused by diarrhoea reduces 85% than control group, and test shows that described lactobacillus salivarius XJP2 is to prevention broiler chicken bacterial diarrhea, reduces mortality ratio and has unusual effect.
Prebiotic effect in broiler chicken body is tested by embodiment 5, lactobacillus salivarius XJP2
Test adopts single factor test completely randomized design, and 135 health AA broiler chicken on the 1st is divided into 3 groups at random, often organizes 3 repetitions, and each repeats 15 chickens. Control group daily ration and drinking-water do not add lactobacillus salivarius, test group daily ration adds 10 respectively6cfu/g��107Cfu/g lactobacillus salivarius, age on the 42nd gets caecum Digesta samples, and real-time fluorescence quantitative PCR measures the total bacterium of sample, lactobacillus, intestinal bacteria bacterium amount; DGGE analyzing total bacterium diversity. Table 3 and the display of table 4 experimental result, add 10 in daily ration7Cfu/g lactobacillus salivarius XJP2, can significantly improve lactic acid bacterium number in caecum (P < 0.05); Improve intestinal microflora diversity (as shown in Figure 3), intestinal microflora is had certain regulating effect.
Table 3 cecal content bacterium number result (Logcfu/g)
The total bacterium diversity index of table 4 caecum
Above embodiment is only in order to illustrate the technical scheme of the present invention, but not is limited; Although with reference to previous embodiment to invention has been detailed description, for the person of ordinary skill of the art, still the technical scheme described in previous embodiment can being modified, or wherein part technology feature is carried out equivalent replacement; And these amendments or replacement, the spirit and scope of the technical scheme not making essence disengaging the present invention of appropriate technical solution claimed.
Claims (7)
1. a strain lactobacillus salivarius XJP2, it is characterised in that: its called after lactobacillus salivarius of classifyingLactobacillussalivarius, its deposit number is CGMCCNo.11386.
2. lactobacillus salivarius XJP2 according to claim 1, it is characterised in that: described lactobacillus salivarius XJP2 is Gram-positive, and bacterium colony is circular, and neat in edge, oyster white, bacterium colony is smooth.
3. lactobacillus salivarius XJP2 according to claim 1, it is characterised in that: after described lactobacillus salivarius XJP2 cultivates 20h, Pfansteihl output is greater than 18.2g/L.
4. lactobacillus salivarius XJP2 according to claim 1, it is characterised in that: acid, cholate are had good tolerance by described lactobacillus salivarius XJP2.
5. lactobacillus salivarius XJP2 according to claim 1 is for the preparation of the application in the micro-ecological additive of broiler breeding.
6. lactobacillus salivarius XJP2 according to claim 5 is for the preparation of the application in the micro-ecological additive of broiler breeding, it is characterised in that: described lactobacillus salivarius XJP2 can significantly improve lactic acid bacterium number in caecum.
7. lactobacillus salivarius XJP2 according to claim 5 is for the preparation of the application in the micro-ecological additive of broiler breeding, it is characterised in that: described lactobacillus salivarius XJP2 can prevent broiler chicken bacterial diarrhea.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610152385.2A CN105624071A (en) | 2016-03-17 | 2016-03-17 | Lactobacillus salivarius XJP2 and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106399197A (en) * | 2016-10-28 | 2017-02-15 | 中国人民解放军疾病预防控制所 | Lactobacillus salivarius and application thereof |
| CN110157645A (en) * | 2019-05-29 | 2019-08-23 | 中国科学技术大学先进技术研究院 | A kind of Lactobacillus salivarius Y4 and its application |
| CN115261287A (en) * | 2022-09-13 | 2022-11-01 | 青岛根源生物技术集团有限公司 | Composite lactobacillus microbial inoculum and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618452A (en) * | 2011-02-01 | 2012-08-01 | 任发政 | Preparation method, composition and application of lactobacillus salivarius and its metabolites |
| CN103289926A (en) * | 2013-05-30 | 2013-09-11 | 中国农业科学院哈尔滨兽医研究所 | Lactobacillus salivarius strain and application thereof |
| CN104195093A (en) * | 2014-09-24 | 2014-12-10 | 中国人民解放军第三军医大学 | Lactobacillus salivarius and pharmaceutical application thereof |
-
2016
- 2016-03-17 CN CN201610152385.2A patent/CN105624071A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102618452A (en) * | 2011-02-01 | 2012-08-01 | 任发政 | Preparation method, composition and application of lactobacillus salivarius and its metabolites |
| CN103289926A (en) * | 2013-05-30 | 2013-09-11 | 中国农业科学院哈尔滨兽医研究所 | Lactobacillus salivarius strain and application thereof |
| CN104195093A (en) * | 2014-09-24 | 2014-12-10 | 中国人民解放军第三军医大学 | Lactobacillus salivarius and pharmaceutical application thereof |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106399197A (en) * | 2016-10-28 | 2017-02-15 | 中国人民解放军疾病预防控制所 | Lactobacillus salivarius and application thereof |
| CN110157645A (en) * | 2019-05-29 | 2019-08-23 | 中国科学技术大学先进技术研究院 | A kind of Lactobacillus salivarius Y4 and its application |
| CN110157645B (en) * | 2019-05-29 | 2021-03-02 | 中国科学技术大学先进技术研究院 | Lactobacillus salivarius Y4 and application thereof |
| CN115261287A (en) * | 2022-09-13 | 2022-11-01 | 青岛根源生物技术集团有限公司 | Composite lactobacillus microbial inoculum and application thereof |
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