CN101380335A - Compound microbial preparation applied for cultivating baby pig in replacement of medicine additive - Google Patents
Compound microbial preparation applied for cultivating baby pig in replacement of medicine additive Download PDFInfo
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- CN101380335A CN101380335A CNA2008101554625A CN200810155462A CN101380335A CN 101380335 A CN101380335 A CN 101380335A CN A2008101554625 A CNA2008101554625 A CN A2008101554625A CN 200810155462 A CN200810155462 A CN 200810155462A CN 101380335 A CN101380335 A CN 101380335A
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Abstract
The invention discloses a compound microbial blend which is applied in the piglet cultivation to replace a drug additive, the compound microbial blend is prepared according to the following steps: a section of the small intestine of a healthy pig which is just slaughtered is taken by sterile operation, the intestinal mucosa is scrapped off by using a sterile blade, sterile water is added for diluting by 10 times, the mixture is stirred for 2min and placed still for 10min, 1ml of supernatant liquid is taken and inoculated into preenrichment medium culture liquid, the constant temperature culture is carried out for 24h at 30 DEG C; 5 percent by weight of inoculation target strains are fermented for 8 hours at 37 DEG C on an MRS culture medium; the target strains after the culture are mixed with probiotic bacteria Xg02 which are preserved in a laboratory to obtain mixed bacteria liquid; the mixed bacteria liquid is added into a liquid seed culture medium according to 5 percent by weight to obtain liquid seeds; the liquid seeds are added in a solid fermentation culture medium according to 12 percent by weight, and the solid powder microbial blend is obtained by drying and smashing at the temperature which is lower than 65 DEG C when the fermentation is finished; the microbial blend is utilized to replace the drug additive in the piglet cultivation, so as to improve the animal resistance, protect the animals from being affected by external harmful bacterial, promote the growth and prevent the diseases.
Description
Technical field
The present invention relates to microbiology, animal husbandry, biotechnology field, be specifically related to a kind of complex micro organism fungicide that is applied to replace in the piglet breed medicated premix.
Background technology
In the gastrointestinal tract of animal, have relatively stable microbiologic population of growing the residence, and quantity is quite big, is about 10
14Individual microorganism is respectively from 400 different bacteria types.Some microorganisms are wherein arranged to keeping the relatively stable of gastrointestinal tract environment and promoting that nutrient substance plays considerable effect in the digestion and the absorption of intestinal.Fuller discovers, the formation of metastable microbiologic population in the intestinal, help animal to improve resistance, avoid the infection of external noxious bacteria, particularly the gastrointestinal of normal healthy animal is infected, germfree animal is than the easy infection disease more of the animal with complete gastrointestinal tract microbiologic population.Because micropopulation is dropped on the special efficacy in the animal gastrointestinal tract, the notion of animal microecology and theory are suggested and are perfect.In animal farming industry, utilize the animal microecology theory, according to the objective law of animal gastrointestinal tract microecosystem, giving treatment in accordance with the tendency of pathological change improves the gastrointestinal microecological environment, sets up the higher microecological balance of Biomass, thereby plays growth promotion, the effect of anti-disease.Its successful Application shows that microbial ecological agent (Probiotics) uses, and has extremely good feeding effect in animal and fowl fodder.The mechanism of action of microbial ecological agent: 1. the antagonism of pair pathogen, probiotic bacteria stops adhering to of pathogen by the sticking site that occupies on the intestinal mucosa, and in intestinal and pathogen competition nutrient, produces organic acid, H
2O
2And other antibiotic substance etc., suppress the growth of pathogenic bacteria breeding, thereby reach antibiotic effect of curing the disease; 2. nutrient is provided, and facilitating digestion absorbs, and improves food conversion ratio; 3. stimulation body's immunological function, enhancing human body immunity power; 4. purification intestinal environment, animal self and many pathogenic bacterium all can produce various noxious substances, some profitable probliotics can stop the synthetic of toxicity amine and ammonia, in and bacteriotoxin.Lactobacillus can produce organic amine and antibiotic substance, reduces pH value and oxidation-reduction potential in the intestinal, helps host's growth promoter and keeps health; 5. alleviate the stress of body, keep the intestinal microecology balance
Summary of the invention
The objective of the invention is to: a kind of complex micro organism fungicide that is applied to replace in the piglet breed medicated premix is provided, utilizes this microbial bacterial agent to replace medicated premix, improve the animal resistance, avoid the infection of external noxious bacteria, growth promotion, anti-disease.
Technical solution of the present invention is that this complex micro organism fungicide prepares according to following steps:
1. from the small intestinal of health pig, separate, the purification strain; The sterile working obtains just slaughters one section about 4cm of health pig small intestinal, with sterile razor blade intestinal mucosa is scraped, and pours 10 times of sterilized water dilutions into, stirs 2min, leaves standstill 10min, gets supernatant 1mL, is inoculated in the pre-enrichment culture medium of 50mL 30 ℃ of constant temperature culture 24h; Wherein, pre-enrichment culture medium is the fresh milk of filling in the glass stoppered bottle;
2. get pre-enrichment liquid 0.1mL, be applied on the 9cm diameter MRS culture medium, the MRS of falling one deck culture medium again in culture dish is under anaerobic cultivated 36h for 30 ℃ then; MRS culture medium wherein: Carnis Bovis seu Bubali cream 10g, peptone 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, Triammonium citrate 2g, dipotassium hydrogen phosphate 2g, tween 80 1mL, magnesium sulfate 0.58g, manganese sulfate 0.25g transfers pH to 6.2～6.4, aquae destillata standardize solution 1000mL, 121 ℃ of sterilization 15min;
3. above-mentioned dilution coating anaerobism is cultivated gained list bacterium colony and select, its streptococcus intermedius, lactobacillus are the purpose strain;
4. on the MRS culture medium, inoculate the purpose strain, 37 ℃ of fermentations of temperature 8 hours with quality 5%;
5. the streptococcus in the 4th step, lactobacillus mix with the probiotic bacteria Xg02 that laboratory is preserved, and obtain mixed bacteria liquid, and three kinds of bacterium mass ratioes are bacillus: coccus: Xg02=1:1:3;
6. mixed bacteria liquid is according to getting liquid seeds in the quality 5% adding liquid seed culture medium, and wherein, each component percentage ratio is in the liquid seed culture medium: peptone 0.85%, Carnis Bovis seu Bubali cream 2.0%, glucose 2.0%, yeast extract 0.7%, liver infusion 5% (v/v); The wherein preparation of liver infusion: the 200g pig liver adds 500g water, boils, and filtered through gauze is got filtrate;
7. liquid seeds adds in the solid fermentation culture medium by quality 12%, and 37 ℃ fermented 20 hours, and fermentation ends is lower than 65 ℃ of drying and crushing and gets the solid powder microbial bacterial agent; Wherein, the solid fermentation culture medium is: Testa oryzae 47.0%, wheat bran 42.5%, glucose 10.5%, initial pH6.5, moisture content 68%.
The present invention has the following advantages: the probiotic bacteria of 1. forming the microorganism fodder microbial inoculum does not produce any interior extracellular toxin, promptly nontoxic, harmless, safely, have no side effect, help promoting colony balance in the body, have and stick performance preferably, with acidproof, the bile tolerance of probiotic bacteria that viable bacteria bodily form formula plays a role, probiotic strain fast growth, long, the easily preservation of survival period; 2. this microbial bacterial agent contains lactobacillus, streptococcus etc., and viable count is greater than 10
9Individual/g, addition is few in piglet is cultured, and builds up resistance, and feeding cost is low.
Description of drawings
Fig. 1 is the piglet growth curve chart.
The specific embodiment
This complex micro organism fungicide prepares according to following concrete steps:
1. from the small intestinal of health pig, separate, the purification strain; The sterile working obtains just slaughters one section about 4cm of health pig small intestinal, with sterile razor blade intestinal mucosa is scraped, and pours 10 times of sterilized water dilutions into, stirs 2min, leaves standstill 10min, gets supernatant 1mL, is inoculated in the pre-enrichment culture medium of 50ml 30 ℃ of constant temperature culture 24h; Wherein, pre-enrichment culture medium is the fresh milk of filling in the glass stoppered bottle;
2. get pre-enrichment liquid 0.1mL, dilution is applied on the 9cm diameter MRS culture medium, then the MRS of falling one deck culture medium again in culture dish, under anaerobic 30 ℃ of cultivation 36h; MRS culture medium wherein: Carnis Bovis seu Bubali cream 10g, peptone 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, Triammonium citrate 2g, dipotassium hydrogen phosphate 2g, tween 80 1mL, magnesium sulfate 0.58g, manganese sulfate 0.25g transfers pH to 6.2～6.4, aquae destillata standardize solution 1000mL, 121 ℃ of sterilization 15min;
3. above-mentioned dilution coating anaerobism is cultivated gained list bacterium colony and select, its streptococcus intermedius, lactobacillus are the purpose strain;
4. on the MRS culture medium, inoculate the purpose strain, 37 ℃ of fermentations of temperature 8 hours with quality 5%;
5. the streptococcus in the 4th step, lactobacillus mix with the probiotic bacteria Xg02 that laboratory is preserved, and obtain mixed bacteria liquid, and three kinds of bacterium mass ratioes are bacillus: coccus: Xg02=1:1:3;
6. mixed bacteria liquid is according to getting liquid seeds in the quality 5% adding liquid seed culture medium, and wherein, each component percentage ratio is in the liquid seed culture medium: peptone 0.85%, Carnis Bovis seu Bubali cream 2.0%, glucose 2.0%, yeast extract 0.7%, liver infusion 5% (v/v); The wherein preparation of liver infusion: the 200g pig liver adds 500g water, boils, and filtered through gauze is got filtrate;
7. liquid seeds adds in the solid fermentation culture medium by quality 12%, and 37 ℃ fermented 20 hours, and fermentation ends is lower than 65 ℃ of drying and crushing and gets the solid powder microbial bacterial agent; Wherein, the solid fermentation culture medium is: Testa oryzae 47.0%, wheat bran 42.5%, glucose 10.5%, initial pH6.5, moisture content 68%.
The selection of experiment piglet: 25 of the healthy ablactational baby pig of the lean meat species that kind is identical, about 8 kilograms of body weight, sex ratio is suitable, and test is carried out in a week after wean, in the test stage of preparation all test pig is carried out conventional immunity and anthelmintic.
The grouping of experiment piglet: all test pig are numbered, are divided into five groups at random, 5 every group, five groups to be numbered 1,2,3,4,5, the 1 groups be matched group, 2,3,4,5 groups is test group.
The concentration proportioning of microbial bacterial agent in daily ration: by 0.5 ‰, 1 ‰, 2 ‰, 4 ‰ ratio is spiked into respectively in the daily ration of 2,3,4,5 groups of piglets by mass ratio (being microbial bacterial agent and the ratio of the quality of daily ration) with microbial bacterial agent.
Feeding method: the basal diet of convention amount antibiotic (use amount of bambermycin in piglet diet is 10mg/kg) is added in the 1st group of piglet supply, does matched group when experiment finishes; Experimental group is fed the daily ration that has added microbial bacterial agent; Every day, feeding twice, respectively at the morning 7:30-8:00 and afternoon 5:00-5:30 feed the equal free choice feeding of all experiment pig, and supply with clean drinking-water at any time.
The relevant information of experiment piglet is collected as table 1,2,3 and shown in Figure 1, and Fig. 1 is a piglet body weight change curve, and table 1 is the weight table at the whole story of piglet, and table 2 is five groups of weight gain of piglets weight factor analysiss of variance table, and table 3 is a price of deed contrast table.
Heavily show the whole story of table 1 piglet:
Table 1 piglet weight at the whole story
Five groups of weight gain of piglets weight list of table 2 factor variance analytical table:
By harmony in the exterior figure as can be known: in piglet diet, add microbial bacterial agent according to different proportion, growth in piglets is had obvious facilitation, and wherein 2 ‰ addition groups are compared with matched group, and average daily gain improves 31.8%, feedstuff-meat ratio has reduced by 15%, and diarrhea rate has reduced by 91.4%.
Claims (1)
1. one kind is applied to the complex micro organism fungicide that piglet replaces medicated premix in culturing, and it is characterized in that this complex micro organism fungicide prepares according to following steps:
(1) separation, purification strain from the small intestinal of health pig; The sterile working obtains just slaughters one section 4cm of health pig small intestinal, with sterile razor blade intestinal mucosa is scraped, and pours 10 times of sterilized water dilutions into, stirs 2min, leaves standstill 10min, gets supernatant 1mL, is inoculated in the pre-enrichment culture medium of 50ml 30 ℃ of constant temperature culture 24h; Wherein, pre-enrichment culture medium is the fresh milk of filling in the glass stoppered bottle;
(2) get pre-enrichment liquid 0.1mL, dilution is applied on the 9cm diameter MRS culture medium, then the MRS of falling one deck culture medium again in culture dish, under anaerobic 30 ℃ of cultivation 36h; MRS culture medium wherein: Carnis Bovis seu Bubali cream 10g, peptone 10g, yeast extract 5g, glucose 20g, sodium acetate 5g, Triammonium citrate 2g, dipotassium hydrogen phosphate 2g, tween 80 1mL, magnesium sulfate 0.58g, manganese sulfate 0.25g transfers pH to 6.2～6.4, aquae destillata standardize solution 1000mL, 121 ℃ of sterilization 15min;
(3) above-mentioned dilution coating anaerobism is cultivated gained list bacterium colony and select, its streptococcus intermedius, lactobacillus are the purpose strain;
(4) on the MRS culture medium, inoculate the purpose strain, 37 ℃ of fermentations of temperature 8 hours with quality 5%;
The streptococcus in (5) the 4th steps, lactobacillus mix with the probiotic bacteria Xg02 that laboratory is preserved, and obtain mixed bacteria liquid, and three kinds of bacterium mass ratioes are bacillus: coccus: Xg02=1:1:3;
(6) mixed bacteria liquid is according to getting liquid seeds in the quality 5% adding liquid seed culture medium, and wherein, each component percentage ratio is in the liquid seed culture medium: peptone 0.85%, Carnis Bovis seu Bubali cream 2.0%, glucose 2.0%, yeast extract 0.7%, liver infusion 5% (v/v); The wherein preparation of liver infusion: the 200g pig liver adds 500g water, boils, and filtered through gauze is got filtrate;
(7) liquid seeds adds in the solid fermentation culture medium by quality 12%, and 37 ℃ fermented 20 hours, and fermentation ends is lower than 65 ℃ of drying and crushing and gets the solid powder microbial bacterial agent; Wherein, the solid fermentation culture medium is: Testa oryzae 47.0%, wheat bran 42.5%, glucose 10.5%, initial pH6.5, moisture content 68%.
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| CNA2008101554625A CN101380335A (en) | 2008-10-06 | 2008-10-06 | Compound microbial preparation applied for cultivating baby pig in replacement of medicine additive |
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| CNA2008101554625A CN101380335A (en) | 2008-10-06 | 2008-10-06 | Compound microbial preparation applied for cultivating baby pig in replacement of medicine additive |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329750A (en) * | 2011-09-21 | 2012-01-25 | 中国科学院亚热带农业生态研究所 | Method for separating pediococcus pentosaceus |
| CN106387317A (en) * | 2016-08-31 | 2017-02-15 | 上海绿博生物科技发展有限公司 | Microorganism feed additive capable of protecting pig livers and preparation method of microorganism feed additive |
-
2008
- 2008-10-06 CN CNA2008101554625A patent/CN101380335A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329750A (en) * | 2011-09-21 | 2012-01-25 | 中国科学院亚热带农业生态研究所 | Method for separating pediococcus pentosaceus |
| CN106387317A (en) * | 2016-08-31 | 2017-02-15 | 上海绿博生物科技发展有限公司 | Microorganism feed additive capable of protecting pig livers and preparation method of microorganism feed additive |
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Open date: 20090311 |