CN104212735A - Poultry composite probiotic preparation and preparation method thereof - Google Patents
Poultry composite probiotic preparation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a poultry composite probiotic preparation and a preparation method thereof. The preparation method comprises the following steps: mixing bacillus subtilis liquid, bacillus licheniformis liquid, enterococcus faecalis liquid, lactobacillus acidophilus liquid, and clostridium butyricum liquid according to a volume ratio of 20-40:5-20:10-30:12-35:15-30, adding carboxymethyl cellulose to prepare suspension, adding the suspension into an aluminum sulfate solution to form agglomerated capsules, filtering to obtain the agglomerated capsulated, washing the capsules with water, and finally drying so as to obtain the poultry composite probiotic preparation; wherein the number of live bacterium in each bacterium liquid is not less than 109 CFU/mL. The composite probiotic preparation with multiple physiological functions is obtained by reasonably compounding the aerobic bacteria and anaerobic bacteria (bacillus subtilis, bacillus licheniformis, enterococcus faecalis, lactobacillus acidophilus, and clostridium butyricum). The provided poultry composite probiotic preparation can modulate the microecological environment in intestines, inhibit the growth of pathogenic bacteria in intestines, strengthen the organism immunity, and increase the feed utilization rate.
Description
Technical field
The present invention relates to a kind of fowl composite probiotics preparations, also relate to the preparation method of said preparation simultaneously, belong to microbial preparation technical field.
Background technology
Along with the development of intensive culture industry, the various diseases of bird constantly occur, and day by day serious.The Main Means of current control poultry disease uses microbiotic, but antibiotic use easily causes the generation of bacterial resistance and the residual of bird product Chinese traditional medicine.Therefore, the disease of prevention bird occurs, and solving the residual of its byproduct Chinese traditional medicine is problem demanding prompt solution.
Probiotic bacterium is the living microorganism that a class is useful to host, can improve the microecological balance in host, plays beneficial effect.The action character of probiotic bacterium forms biological protection film to prevent pathogenic bacterium from invading; Produce the material of lactic acid or acetic acid, hydrogen peroxide, bacteriocin and similar antibiotic, and suppress the growth of pathogenic bacteria with this, adjustment host immune response, improve the activity that digestive tube bacterial metabolism disease reduces harmful enzyme in excrement; Reduce the incidence etc. of bacterial translocation, therefore probiotic bacterium has the barrier function or enhancement non-specific immune function of strengthening enteric microorganism region, regulates specific immune function, strengthens disease resistance and physique, control courses of infection, improves the function of efficiency of feed utilization and growth of animal rate.Therefore arrange in pairs or groups to different probiotic bacterium, the effect improving them becomes study hotspot.
Summary of the invention
The object of this invention is to provide a kind of fowl composite probiotics preparations, can the micro-ecological environment of regulating intestinal canal, suppress the growth of enteric pathogenic bacteria, enhancing body immunizing power, improve efficiency of feed utilization.
The present invention also aims to the preparation method that a kind of fowl composite probiotics preparations is provided.
In order to realize above object, the technical solution adopted in the present invention is to provide a kind of fowl composite probiotics preparations, by the 20-40:5-20:10-30:12-35:15-30 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filter and obtain condensing capsule, be drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml.
In described fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
The technical solution adopted in the present invention is also the preparation method providing a kind of fowl composite probiotics preparations, comprise the following steps: subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid are mixed by volume, then the carboxymethyl cellulose adding mixed bacteria liquid quality 0.5-1% makes suspension, again suspension is added in the alum liquor of suspension volume 1-2 massfraction 3-5% doubly, form cohesion capsule, filtration obtains condensing capsule, washing, drying obtains fowl composite probiotics preparations; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml.
The preparation method of described subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid comprises the following steps:
(1) activation of bacterial classification: subtilis, Bacillus licheniformis, clostridium butyricum are inoculated in nutrient broth medium respectively, 37 DEG C, cultivate 12-14h under 180-200r/min condition;
(2) with 3% inoculum size, the subtilis of activation, Bacillus licheniformis, clostridium butyricum are accessed liquid nutrient medium respectively, under 37 DEG C of conditions, logical aerobe fermentation cultivates 36-48h, filter then by centrifugal for fermented liquid 3000-4000r/m 5-10min, remove supernatant liquor, add sterilized water and obtain subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
Described liquid nutrient medium comprises the component of following massfraction: glucose 0.1%, starch 0.2%, soybean meal hydrolysate 2%, potassium primary phosphate 0.3%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.05%, iron(ic) chloride 0.01%, calcium carbonate 0.01%.
The preparation method of described enterococcus faecalis bacterium liquid comprises the following steps:
(1) activation of bacterial classification: enterococcus faecalis is inoculated in nutrient broth medium, static gas wave refrigerator 12-14h under 37 DEG C of conditions;
(2) with 2% inoculum size, the enterococcus faecalis of activation is accessed MRS substratum, under 37 DEG C of conditions, standing for fermentation cultivates 24-36h, then by centrifugal for fermented liquid 3000-4000r/m 5-10min, removes supernatant liquor, add sterilized water and obtain enterococcus faecalis bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
Described MRS substratum comprises the component of following massfraction: the leaching of Tryptones 1%, yeast powder 0.5%, glucose 1%, sucrose 0.5%, sodium-acetate 1.5%, ammonium citrate 0.2%, potassium primary phosphate 0.4%, manganous sulfate 0.05%, magnesium sulfate 0.02%, ferric sulfate 0.003%, tween 80 0.1%.
The preparation method of described Lactobacterium acidophilum bacterium liquid comprises the following steps:
(1) activation of bacterial classification: Lactobacterium acidophilum is inoculated in nutrient broth medium, static gas wave refrigerator 12-14h under 37 DEG C of conditions;
(2) with 2% inoculum size, the Lactobacterium acidophilum of activation is accessed fermention medium, under 37 DEG C of conditions, standing for fermentation cultivates 24-36h, then by centrifugal for fermented liquid 3000-4000r/m 5-10min, remove supernatant liquor, add sterilized water and obtain Lactobacterium acidophilum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
Described fermention medium comprises the component of following massfraction: the leaching of glucose 3%, lactose 1%, skimmed milk powder 5%, yeast powder 0.8%, potassium primary phosphate 0.3%, Sodium phosphate dibasic 0.4%, magnesium sulfate 0.02%, manganous sulfate 0.008%.
Described nutrient broth medium comprises the component of following massfraction: peptone 1%, extractum carnis 0.3%, sodium-chlor 0.5%.
The present invention by by the aerobic bacterias such as subtilis, Bacillus licheniformis, enterococcus faecalis, Lactobacterium acidophilum, clostridium butyricum and anerobe reasonably combined, obtain the composite probiotics preparations with different physiological roles.Fowl of the present invention can the micro-ecological environment of regulating intestinal canal with composite probiotics preparations, suppresses the growth of enteric pathogenic bacteria, enhancing body immunizing power, improves efficiency of feed utilization.
Fowl of the present invention composite probiotics preparations can preserve 1 year at normal temperatures.During use, by fowl with composite probiotics preparations with 1% weight fraction add to feed or drinking-water in.
Embodiment
The preparation of substratum
Nutrient broth medium comprises the component of following massfraction: peptone 1%, extractum carnis 0.3%, sodium-chlor 0.5%; PH7-7.2.
Liquid nutrient medium comprises the component of following massfraction: glucose 0.1%, starch 0.2%, soybean meal hydrolysate 2%, potassium primary phosphate 0.3%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.05%, iron(ic) chloride 0.01%, calcium carbonate 0.01%; PH7-7.2.
MRS substratum comprises the component of following massfraction: the leaching of Tryptones 1%, yeast powder 0.5%, glucose 1%, sucrose 0.5%, sodium-acetate 1.5%, ammonium citrate 0.2%, potassium primary phosphate 0.4%, manganous sulfate 0.05%, magnesium sulfate 0.02%, ferric sulfate 0.003%, tween 80 0.1%; PH6.5-7.
Fermention medium comprises the component of following massfraction: the leaching of glucose 3%, lactose 1%, skimmed milk powder 5%, yeast powder 0.8%, potassium primary phosphate 0.3%, Sodium phosphate dibasic 0.4%, magnesium sulfate 0.02%, manganous sulfate 0.008%; PH5.7-6.
Embodiment 1
The fowl composite probiotics preparations of the present embodiment, by the 20:10:20:12:30 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filtration obtains condensing capsule, is drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml; In fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
The preparation method of the fowl composite probiotics preparations of the present embodiment, comprises the following steps:
1, the preparation of bacterium liquid
(1) preparation of subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid
The activation of (a) bacterial classification: subtilis, Bacillus licheniformis, clostridium butyricum are inoculated in nutrient broth medium respectively, 37 DEG C, cultivate 12h under 200r/min condition;
B the subtilis of activation, Bacillus licheniformis, clostridium butyricum are accessed liquid nutrient medium with 3% inoculum size by () respectively, under 37 DEG C of conditions, logical aerobe fermentation cultivates 36h, then by centrifugal for fermented liquid 4000r/m 5min after filtration, remove supernatant liquor, add sterilized water and obtain subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(2) preparation of enterococcus faecalis bacterium liquid
The activation of (a) bacterial classification: enterococcus faecalis is inoculated in nutrient broth medium, static gas wave refrigerator 12h under 37 DEG C of conditions;
B the enterococcus faecalis of activation is accessed MRS substratum with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 24h, then by centrifugal for fermented liquid 4000r/m 5min, removes supernatant liquor, add sterilized water and obtain enterococcus faecalis bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(3) preparation of Lactobacterium acidophilum bacterium liquid
The activation of (a) bacterial classification: Lactobacterium acidophilum is inoculated in nutrient broth medium, static gas wave refrigerator 12h under 37 DEG C of conditions;
B the Lactobacterium acidophilum of activation is accessed fermention medium with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 24h, then by centrifugal for fermented liquid 4000r/m 5min, removes supernatant liquor, add sterilized water and obtain Lactobacterium acidophilum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
2, subtilis, Bacillus licheniformis, enterococcus faecalis, Lactobacterium acidophilum, clostridium butyricum are mixed in proportion, then the carboxymethyl cellulose adding mixed bacteria liquid quality 1% makes suspension, again suspension is added in the alum liquor of massfraction 5% of suspension volume 1 times, form cohesion capsule, filtration obtains condensing capsule, washing, 42 DEG C of dryings obtain fowl composite probiotics preparations.
Embodiment 2
The fowl composite probiotics preparations of the present embodiment, by the 30:20:10:20:15 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filtration obtains condensing capsule, is drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml; In fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
The preparation method of the fowl composite probiotics preparations of the present embodiment, comprises the following steps:
1, the preparation of bacterium liquid
(1) preparation of subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid
The activation of (a) bacterial classification: subtilis, Bacillus licheniformis, clostridium butyricum are inoculated in nutrient broth medium respectively, 37 DEG C, cultivate 14h under 180r/min condition;
B the subtilis of activation, Bacillus licheniformis, clostridium butyricum are accessed liquid nutrient medium with 3% inoculum size by () respectively, under 37 DEG C of conditions, logical aerobe fermentation cultivates 48h, then by centrifugal for fermented liquid 3000r/m 10min after filtration, remove supernatant liquor, add sterilized water and obtain subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(2) preparation of enterococcus faecalis bacterium liquid
The activation of (a) bacterial classification: enterococcus faecalis is inoculated in nutrient broth medium, static gas wave refrigerator 14h under 37 DEG C of conditions;
B the enterococcus faecalis of activation is accessed MRS substratum with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 36h, then by centrifugal for fermented liquid 3000r/m 10min, removes supernatant liquor, add sterilized water and obtain enterococcus faecalis bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(3) preparation of Lactobacterium acidophilum bacterium liquid
The activation of (a) bacterial classification: Lactobacterium acidophilum is inoculated in nutrient broth medium, static gas wave refrigerator 14h under 37 DEG C of conditions;
B the Lactobacterium acidophilum of activation is accessed fermention medium with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 36h, then by centrifugal for fermented liquid 3000r/m 10min, removes supernatant liquor, add sterilized water and obtain Lactobacterium acidophilum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
2, subtilis, Bacillus licheniformis, enterococcus faecalis, Lactobacterium acidophilum, clostridium butyricum are mixed in proportion, then the carboxymethyl cellulose adding mixed bacteria liquid quality 0.5% makes suspension, again suspension is added in the alum liquor of massfraction 4% of suspension volume 1.5 times, form cohesion capsule, filtration obtains condensing capsule, washing, 60 DEG C of dryings obtain fowl composite probiotics preparations.
Embodiment 3
The fowl composite probiotics preparations of the present embodiment, by the 40:5:30:35:25 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filtration obtains condensing capsule, is drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml; In fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
The preparation method of the fowl composite probiotics preparations of the present embodiment, comprises the following steps:
1, the preparation of bacterium liquid
(1) preparation of subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid
The activation of (a) bacterial classification: subtilis, Bacillus licheniformis, clostridium butyricum are inoculated in nutrient broth medium respectively, 37 DEG C, cultivate 13h under 190r/min condition;
B the subtilis of activation, Bacillus licheniformis, clostridium butyricum are accessed liquid nutrient medium with 3% inoculum size by () respectively, under 37 DEG C of conditions, logical aerobe fermentation cultivates 42h, then by centrifugal for fermented liquid 3500r/m 8min after filtration, remove supernatant liquor, add sterilized water and obtain subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(2) preparation of enterococcus faecalis bacterium liquid
The activation of (a) bacterial classification: enterococcus faecalis is inoculated in nutrient broth medium, static gas wave refrigerator 13h under 37 DEG C of conditions;
B the enterococcus faecalis of activation is accessed MRS substratum with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 30h, then by centrifugal for fermented liquid 3500r/m 8min, removes supernatant liquor, add sterilized water and obtain enterococcus faecalis bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
(3) preparation of Lactobacterium acidophilum bacterium liquid
The activation of (a) bacterial classification: Lactobacterium acidophilum is inoculated in nutrient broth medium, static gas wave refrigerator 13h under 37 DEG C of conditions;
B the Lactobacterium acidophilum of activation is accessed fermention medium with 2% inoculum size by (), under 37 DEG C of conditions, standing for fermentation cultivates 30h, then by centrifugal for fermented liquid 3500r/m 8min, removes supernatant liquor, add sterilized water and obtain Lactobacterium acidophilum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml;
2, subtilis, Bacillus licheniformis, enterococcus faecalis, Lactobacterium acidophilum, clostridium butyricum are mixed in proportion, then the carboxymethyl cellulose adding mixed bacteria liquid quality 0.8% makes suspension, again suspension is added in the alum liquor of massfraction 3% of suspension volume 2 times, form cohesion capsule, filtration obtains condensing capsule, washing, 50 DEG C of dryings obtain fowl composite probiotics preparations.
Comparative example
The fowl composite probiotics preparations of this comparative example, by the 40:5:30:35 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filtration obtains condensing capsule, is drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml; In fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
Experimental example 1
The preservation period test of fowl composite probiotics preparations of the present invention
Placed 12 months under 25 DEG C of conditions by fowl of the present invention composite probiotics preparations, and observed outward appearance respectively and measured viable count 0,1,2,4,8,12 month time, result is as shown in table 1.
The preservation period test of table 1 fowl composite probiotics preparations of the present invention
Test-results shows, composite probiotics preparations of the present invention is active basicly stable at normal temperatures.
Experimental example 2
The effect analysis of fowl composite probiotics preparations of the present invention
This experimental example chicken used basal diet is commercially available conventional feed.
Experimental animal: choose the chick 200 be in a good state of health, be divided into 5 groups at random, often organizes 40.
Test group: test group 1-3 group addition of feeding respectively is the chicken basal diet of the composite probiotics preparations of 1% embodiment 1-3, comparative group addition of feeding is the chicken basal diet of the composite probiotics preparations of 1% comparative example, blank group of chicken basal diet of feeding.
Test method: take to raise in cages in house, free choice feeding, drinking-water, each amount of feeding have enough with chicken after in hopper slightly clout be degree, feed continuously 30 days.Duration of test, the feeding coal of heavy, the feed of accurate recording chicken every day and spreading amount, and the 30th day morning of test, each selection 10 healthy chickens that repeat carry out the collection of excrement sample, gather fresh pollution-free ight soil with 50ml sterile centrifugation tube, put into-20 DEG C of Refrigerator stores to be measured.
Observe the impact of livestock and poultry of the present invention composite probiotics preparations on broiler growth performance and Immune Organs Index, test-results refers to following table 2, table 3, table 4.
The each treatment group of table 2 is on the impact of chicken growth performance
| Group | Day weight gain (g/d) | Daily ingestion amount (g/d) | Feed-weight ratio |
| Test group 1 | 39.45±0.68 | 83.26±3.48 | 2.11±0.02 |
| Test group 2 | 39.17±0.82 | 82.67±10.39 | 2.12±0.03 |
| Test group 3 | 40.54±0.51 | 83.09±2.27 | 2.04±0.07 |
| Comparative group | 36.12±0.86 | 81.95±2.29 | 2.26±0.05 |
| Blank group | 35.85±0.76 | 81.15±5.36 | 2.30±0.08 |
The each treatment group of table 3 is on the impact of Immune Organs of Chicken
| Group | Thymus index (mg/g) | Spleen index (mg/g) | Bursal index (mg/g) |
| Test group 1 | 6.98±1.45 | 2.19±0.56 | 5.98±1.75 |
| Test group 2 | 7.12±1.63 | 2.21±0.69 | 5.89±1.58 |
| Test group 3 | 7.34±2.00 | 2.31±0.76 | 6.12±1.34 |
| Comparative group | 6.81±1.66 | 1.98±0.36 | 5.58±1.27 |
| Blank group | 6.40±1.50 | 1.84±0.27 | 4.76±1.20 |
The each treatment group of table 4 on chicken manure just in the impact (the fresh content of log10CFU/g) of microorganism
| Project | Test group 1 | Test group 2 | Test group 3 | Comparative group | Blank group |
| Intestinal bacteria | 3.45±1.65 | 4.12±0.98 | 3.24±0.76 | 4.51±1.23 | 6.73±0.85 |
| Salmonellas | 0.75±2.01 | 0.63±1.68 | 0.58±1.29 | 0.88±1.56 | 1.72±2.00 |
Found out by table 2,3 and 4, the composite probiotics preparations of embodiment 1-3 is regulating chicken growth performance, is strengthening the composite probiotics preparations being all better than comparative group in immunologic function and resistance against diseases.
Claims (10)
1. a fowl composite probiotics preparations, it is characterized in that, by the 20-40:5-20:10-30:12-35:15-30 mixing by volume of subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid, then add carboxymethyl cellulose and make suspension, again suspension is added in alum liquor, form cohesion capsule, filter and obtain condensing capsule, be drying to obtain after washing; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml.
2. fowl composite probiotics preparations according to claim 1, is characterized in that, in described fowl composite probiotics preparations, total viable count is not less than 5 × 10
9cFU/g.
3. the preparation method of a fowl composite probiotics preparations as claimed in claim 1, it is characterized in that, comprise the following steps: subtilis bacterium liquid, Bacillus licheniformis liquid, enterococcus faecalis bacterium liquid, Lactobacterium acidophilum bacterium liquid, clostridium butyricum bacterium liquid are mixed by volume, then the carboxymethyl cellulose adding mixed bacteria liquid quality 0.5-1% makes suspension, again suspension is added in the alum liquor of suspension volume 1-2 massfraction 3-5% doubly, form cohesion capsule, filtration obtains condensing capsule, washing, drying obtains fowl composite probiotics preparations; Viable count in above-mentioned bacterial classification bacterium liquid is all not less than 10
9cFU/ml.
4. the preparation method of fowl composite probiotics preparations according to claim 3, is characterized in that, the preparation method of described subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid comprises the following steps:
(1) activation of bacterial classification: subtilis, Bacillus licheniformis, clostridium butyricum are inoculated in nutrient broth medium respectively, 37 DEG C, cultivate 12-14h under 180-200r/min condition;
(2) with 3% inoculum size, the subtilis of activation, Bacillus licheniformis, clostridium butyricum are accessed liquid nutrient medium respectively, under 37 DEG C of conditions, logical aerobe fermentation cultivates 36-48h, filter then by centrifugal for fermented liquid 3000-4000r/m 5-10min, remove supernatant liquor, add sterilized water and obtain subtilis, Bacillus licheniformis, clostridium butyricum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
5. the preparation method of fowl composite probiotics preparations according to claim 4, it is characterized in that, described liquid nutrient medium comprises the component of following massfraction: glucose 0.1%, starch 0.2%, soybean meal hydrolysate 2%, potassium primary phosphate 0.3%, dipotassium hydrogen phosphate 0.2%, magnesium sulfate 0.05%, iron(ic) chloride 0.01%, calcium carbonate 0.01%.
6. the preparation method of fowl composite probiotics preparations according to claim 3, is characterized in that, the preparation method of described enterococcus faecalis bacterium liquid comprises the following steps:
(1) activation of bacterial classification: enterococcus faecalis is inoculated in nutrient broth medium, static gas wave refrigerator 12-14h under 37 DEG C of conditions;
(2) with 2% inoculum size, the enterococcus faecalis of activation is accessed MRS substratum, under 37 DEG C of conditions, standing for fermentation cultivates 24-36h, then by centrifugal for fermented liquid 3000-4000r/m 5-10min, removes supernatant liquor, add sterilized water and obtain enterococcus faecalis bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
7. the preparation method of fowl composite probiotics preparations according to claim 6, it is characterized in that, described MRS substratum comprises the component of following massfraction: the leaching of Tryptones 1%, yeast powder 0.5%, glucose 1%, sucrose 0.5%, sodium-acetate 1.5%, ammonium citrate 0.2%, potassium primary phosphate 0.4%, manganous sulfate 0.05%, magnesium sulfate 0.02%, ferric sulfate 0.003%, tween 80 0.1%.
8. the preparation method of fowl composite probiotics preparations according to claim 3, is characterized in that, the preparation method of described Lactobacterium acidophilum bacterium liquid comprises the following steps:
(1) activation of bacterial classification: Lactobacterium acidophilum is inoculated in nutrient broth medium, static gas wave refrigerator 12-14h under 37 DEG C of conditions;
(2) with 2% inoculum size, the Lactobacterium acidophilum of activation is accessed fermention medium, under 37 DEG C of conditions, standing for fermentation cultivates 24-36h, then by centrifugal for fermented liquid 3000-4000r/m 5-10min, remove supernatant liquor, add sterilized water and obtain Lactobacterium acidophilum bacterium liquid, in gained bacterium liquid, viable count is all not less than 10
9cFU/ml.
9. the preparation method of fowl composite probiotics preparations according to claim 8, it is characterized in that, described fermention medium comprises the component of following massfraction: the leaching of glucose 3%, lactose 1%, skimmed milk powder 5%, yeast powder 0.8%, potassium primary phosphate 0.3%, Sodium phosphate dibasic 0.4%, magnesium sulfate 0.02%, manganous sulfate 0.008%.
10. the preparation method of the fowl composite probiotics preparations according to claim 4,6 or 8, is characterized in that, described nutrient broth medium comprises the component of following massfraction: peptone 1%, extractum carnis 0.3%, sodium-chlor 0.5%.
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| CN105707478A (en) * | 2016-03-11 | 2016-06-29 | 山东鹤来生物科技有限公司 | Composite probiotic culture and preparation method thereof |
| CN106615626A (en) * | 2017-01-10 | 2017-05-10 | 武汉新联大生物有限公司 | Probiotic dry suspension for feed and preparation method thereof |
| CN106801013A (en) * | 2015-11-26 | 2017-06-06 | 瑞普（天津）生物药业有限公司 | One kind mixing probiotics dried object and preparation method |
| CN107254424A (en) * | 2017-07-11 | 2017-10-17 | 福建贝迪药业有限公司 | A kind of livestock and poultry new liquid compound micro-ecological preparation and preparation method thereof |
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| CN106801013A (en) * | 2015-11-26 | 2017-06-06 | 瑞普（天津）生物药业有限公司 | One kind mixing probiotics dried object and preparation method |
| CN105707478A (en) * | 2016-03-11 | 2016-06-29 | 山东鹤来生物科技有限公司 | Composite probiotic culture and preparation method thereof |
| CN106615626A (en) * | 2017-01-10 | 2017-05-10 | 武汉新联大生物有限公司 | Probiotic dry suspension for feed and preparation method thereof |
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| CN107254424B (en) * | 2017-07-11 | 2023-02-28 | 福建贝迪药业有限公司 | Novel liquid composite microecological preparation for livestock and poultry and preparation method thereof |
| CN108782945A (en) * | 2018-04-19 | 2018-11-13 | 长沙道勤生物科技有限公司 | A kind of feed addictive of layer chicken preparation method that laying rate can be improved |
| CN112210505A (en) * | 2019-07-12 | 2021-01-12 | 陆良佰佳益康生物科技有限公司 | Preparation method of efficient composite probiotics for poultry intestinal tracts |
| CN113575758A (en) * | 2021-08-04 | 2021-11-02 | 广西大学 | Composite probiotics and preparation method thereof |
| CN114272357B (en) * | 2021-12-30 | 2022-11-01 | 江苏三仪生物工程有限公司 | Biological compound preparation for waterfowl for preventing and treating diarrhea and reducing feed conversion ratio |
| CN114272357A (en) * | 2021-12-30 | 2022-04-05 | 江苏三仪生物工程有限公司 | Biological compound preparation for waterfowl for preventing and treating diarrhea and reducing feed conversion ratio |
| CN114854651B (en) * | 2022-07-06 | 2022-09-30 | 广东顺德鼎一生物科技有限公司 | Micrococcus compositus preparation and preparation method and application thereof |
| CN114854651A (en) * | 2022-07-06 | 2022-08-05 | 广东顺德鼎一生物科技有限公司 | Micrococcus compositus preparation and preparation method and application thereof |
| CN115851543A (en) * | 2022-12-19 | 2023-03-28 | 青岛慧和生物科技有限公司 | Biological activator for promoting development of avian embryo |
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