CN103181460A

CN103181460A - Preparation method and application of lactobacillus plantarum for feed

Info

Publication number: CN103181460A
Application number: CN2011104479153A
Authority: CN
Inventors: 莫少华; 张江; 王晓娟
Original assignee: BEIJING RESOURCE ASIA-PACIFIC FEED TECHNOLOGY Co Ltd
Current assignee: BEIJING RESOURCE ASIA-PACIFIC FEED TECHNOLOGY Co Ltd
Priority date: 2011-12-27
Filing date: 2011-12-27
Publication date: 2013-07-03

Abstract

The invention provides a preparation method for lactobacillus plantarum used for a feed. To overcome the problems of abuse of antibiotics in farming, generation of drug resistance and the like at present, application of a novel antibiotic substitute product, i.e., probiotics, is a trend of modern farming. The probiotics is prepared through deep liquid state fermentation, has the advantages of low production cost and a high viable count and is a stable, highly-efficient and green feed additive.

Description

A kind of preparation method of forage plant lactobacillus and application thereof

Technical field

The present invention relates to a kind of preparation method and application of green feed additive probiotics Lactobacillus plantarum.

Technical background

Antibioticly be applied in animal husbandry and produced huge economic benefit.Investigation shows that the application of antibiotic in animal husbandry can make pork output improve 4%.But the negative effect that antibiotic a large amount of use is followed also becomes increasingly conspicuous, and it is residual in animal body mainly to show as the chemical sproof generation of pathogenic microorganism and medicine, thereby human beings'health is produced direct or potential harm.The application of antibiotic in feed runs into stern challenge.Therefore the research and development of antibiotic substitute (Lactobacillus plantarum and acid supplement etc.) become the focus of green feed additive in recent years, and wherein the application of Lactobacillus plantarum receives much concern.Lactobacillus plantarum is the bacterial classification that Ministry of Agriculture's approval can be used for feed addictive, the effect that it is characterized in that producing acid, produces enzyme and inhibition enteric pathogenic bacteria.Lactobacillus plantarum improves the production performance of animal by the micro-ecological environment that improves animal intestinal and the immunity that improves animal, and noresidue in animal product.But the method that adopts traditional carbon nitrogen source optimization can make Lactobacillus plantarum reach 10 ⁹Behind the CFU/ml, improve again and be difficult to behind the carbon nitrogen source content further improve, illustrate that the conventional carbon nitrogenous source has satisfied its growth.Here add prebiotics 0.1% wood oligose and 0.1% few fructose that can promote the Lactobacillus plantarum growth in zymotic fluid after, its zymotic fluid viable bacteria density of fermenting brings up to 10 ¹⁰CFU/ml, zoopery simultaneously shows that the forage plant lactobacillus can well improve growth performance.

Summary of the invention

The object of the invention is the antibiotic substitute invented at current antibiotic abuse, the noresidue of this product, and nonhazardous is the novel fodder additive of green health.

The invention provides a kind of preparation method for probiotics, it comprises the steps:

1) preparation seed culture medium 500ml sterilized 15 minutes for 121 ℃;

2) with corresponding frozen bacterial classification inoculation and seed culture medium, static cultivation 18 hours.

3) preparation fermentation medium 500L sterilized 15 minutes for 121 ℃;

4) with 2) cultivate in the seed adding fermentation tank that obtains, fermented 24 hours;

5) after the fermentation ends, collect zymotic fluid, add 50% zeolite powder, 50% powdered rice hulls low temperature drying, the compound micro-ecological preparation of namely making after each single bacterium mixes.

Below technology contents of the present invention is described further.

1) lactic acid bacteria seed culture medium is MRS meat soup.

2) the lactobacillus-fermented medium component is as follows

Lactic acid bacteria is mended ammoniacal liquor and makes the pH value keep about 7.0 during the fermentation.

Raw material	Ratio	500L consumption (kg)
			Brown sugar	3％	15
Few wood sugar	0.1％	0.5
			Few fructose	0.1％	0.5
Dregs of beans	1％	5
			Dusty yeast	1％	5
Magnesium sulfate	0.05％	2.5
			Manganese sulfate	0.05％	2.5

Gemma bacteria fermentation culture medium composition is as follows:

After fermentation stopped, the Lactobacillus plantarum viable count was 10 ¹⁰CFU/m improves an order of magnitude than not adding few fructose and few wood sugar.

Fermentation costs of the present invention is cheap, and raw material is low-cost raw material and industrial wastes such as molasses, brown sugar, corn flour, dregs of beans and dusty yeast.Fermentation period is 36-42 hour, and the cycle is short, and it is low to consume energy.

This probiotics can use in piglet, growing-finishing pig, bird and other cultivated animals, and addition is as shown in the table:

Type of animal	Addition %	The g/ ton
			Piglet	0.05	500
Growing-finishing pig	0.02-0.05	200-500
			Poultry such as chicken and duck	0.02-0.05	200-500
Milk beef etc.	0.05-0.1	500-1000
			Special animalses such as quail	0.01-0.03	100-500

By the feed addictive of the safety of a kind of green in compound micro-ecological preparation chamber of the present invention preparation, be that mode with bacterium bacterium processed solves cultivated animals diarrhoea, death and growth performance decline and causes degradation problem under the culture benefit.

Further specify the present invention below in an embodiment.

The viable count (figure sees Figure of description) of embodiment 1 Lactobacillus plantarum sweat

Lactobacillus plantarum zymotic fluid viable count (figure sees Figure of description)

Embodiment 2 Lactobacillus plantarums are in the application of weanling pig

1. material and method

1.1 animal used as test and grouping: tri-crossbreeding (male parent: Du Luoke, female parent: grow up), 40 of 28 age in days weanling pigs are divided into two groups at random, every group of 2 repetitions, each repeats 10, male and female half and half.Add the 1000ppm Lactobacillus plantarum in the test group baby pig feedstuff respectively; Control group is blank daily ration.Feed forms for premix (the not containing antibiotic) autogamy of employing 4%, and prescription adopts resource prescription teacher software development

1.2 test products Lactobacillus plantarum

1.3 test period and place

Test period: on July 9th, 1 2010 on June 9th, 2010

Test site: resource Asia-Pacific, Beijing boar Co., Ltd

1.4 test method

Feeding and management: colony house is raised, and free choice feeding freely drink water, and ventilation is good, the program inoculation of preventing epidemic routinely, and day encloses 2 times clearly.

Test data sheet: be responsible for by the special messenger in experimental period, conscientiously record test pig food consumption situation, body weight etc.

1.5 data are handled and are analyzed

This test for data result carries out statistical analysis with the one-way analysis of variance method in the SPSS12.0 statistical software, if use duncan's method (Ducan) to carry out multiple ratio again through F check significant difference person.

2, result and discussion

2.1 result of the test sees the following form

Annotate: duration of test does not have major disease to be taken place

The identical person of shoulder mark lowercase represents difference not remarkable (P＞0.05) in the same row; The complete different persons of lowercase represent significant difference (P＜0.05).

2.2 interpretation of result

2.2.1 average feed intake aspect, test group improves 1.8% than control group, and difference is remarkable (P＞0.05) not.

2.2.2 the average weight gain aspect, test group has improved 6.4% than control group, significant difference (p＜0.05);

2.2.3 the feedstuff-meat ratio aspect, test group has reduced by 4.3% than control group, significant difference (p＜0.05).

Presentation of results, Lactobacillus plantarum can better promote piglet growth.

The application of embodiment 3 Lactobacillus plantarums in poultry

1. material and method

1.1 experimental animal and group experiment animal and daily ration test chicken with 1 age in days AA commodity generations meat male and female mix chick ' 1000; The test daily ration is with reference to national research council standard (NRC, 1994) and AA fowl raising standard, autogamy feed.

1.2 test products Lactobacillus plantarum

1000 tests are divided into 5 groups at random with AA broiler chicken chicken, 200 every group, 5 repetitions, each repeats 40, freely drinks water and searches for food.Control group: blank group, the basal diet of feeding; Antibiotic group: basal diet+antibiotic (10mg/kg): add 0,0.01%, 0.05%, 0.1% Lactobacillus plantarum group in the basal diet respectively.

2, result and discussion

2.1 result of the test sees the following form

Annotate: duration of test does not have major disease to be taken place

2.2 interpretation of result

The production performance of livestock and poultry comprises weightening finish, feed efficiency and survival rate, and this is the important indicator of reflection feed addictive validity, security and economy.Last watch test shows, the survival rate of blank group chicken significantly is lower than other each groups (p＜0.05), and 0.05% group and 0.1% group is significantly higher than the antibiotic group, blank group and 0.01% group (p＜0.05), the average live body of 0.05% group and 0.1% group 42 age in days weighs and weightening finish all is significantly higher than other each groups (p＜0.05), average total augment weight increases by 6.6% (119.1g/ only) and 7.0% (127.4g/ only), significant difference (p＜0.05) respectively than blank group.Lactobacillus plantarum is bigger to the improvement amplitude of feed efficiency, wherein, adds 0.01,0.05 0.1% than blank group group raising 8.8%, 10.5% and 11.6%, difference is (p＜0.05) extremely significantly, and wherein 0.05% group and 0.1% group is significantly higher than the antibiotic group.From every index, Lactobacillus plantarum adds 0.1% group＞0.05% group＞0.01 group＞antibiotic group＞blank group, illustrate that Lactobacillus plantarum can significantly improve efficiency of feed utilization (p＜0.05), weightening finish and survival rate (p＜0.05) by the utmost point, the effect of adding Lactobacillus plantarum is better than antibiotic.

Description of drawings

Fig. 1 is lactobacillus plantarum, microscopically Lactobacillus plantarum form.

Fig. 2 is: the viable count of embodiment 1 Lactobacillus plantarum sweat

Fig. 3 is: Lactobacillus plantarum zymotic fluid viable count.

Claims

1. the invention provides a kind of method of the preparation for the forage plant lactobacillus, it comprises the steps:

1) preparation seed culture medium 500ml sterilized 15 minutes for 121 ℃;

2) with corresponding frozen bacterial classification inoculation and seed culture medium, the static cultivation of Lactobacillus plantarum 18 hours,

3) preparation fermentation medium 500L, add 0.5% increase the bacteria factor FOS, 121 ℃ of sterilizations 15 minutes;

4) with 2) cultivate in the seed adding fermentation tank that obtains, fermented about 24 hours;

2. according to the preparation method of claim 1, wherein said lactobacillus-fermented culture medium is brown sugar 3%, dregs of beans 1%, dusty yeast 1%, FOS 0.5%, magnesium sulfate 0.05%, manganese sulfate 0.05%.

3. preparation method according to claim 1, wherein said lactic acid bacteria seed culture medium is MRS meat soup.

4. preparation method according to claim 1, wherein said complex microorganism preparations comprises Lactobacillus plantarum and metabolite thereof.

5. preparation method according to claim 1 is characterized in that step 4) is judged that the lactobacillus-fermented terminal point no longer descends 2 for pH after mending alkali lye) adding of FOS can obviously improve the viable count of Lactobacillus plantarum zymotic fluid.

6. the preparation method of a feeding micro-ecological preparation, its method according to claim 1-5 is prepared from.