CN104489268A - Micro-ecological preparation and preparation method thereof - Google Patents
Micro-ecological preparation and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a micro-ecological preparation. The micro-ecological preparation is a beta-cyclodextrin clathrate compound and provides a preparation method of the micro-ecological preparation. The micro-ecological preparation has the beneficial effects that beta-cyclodextrin has a unique internally-lyophobic and externally-hydrophilic structure and can form a clathrate compound with a plurality of moderate-sized guest molecules; the beta-cyclodextrin is capable of covering probiotics to form a relatively-stable clathrate compound; the clathrate compound can be used for effectively protecting the compound from oxidation reaction or hydrolysis reaction; meanwhile, the stability of the medicine is also improved; the water solubility is improved; the release of the micro-ecological preparation is controlled; the bioavailability is improved.
Description
Technical field
The present invention relates to field of veterinary, be specifically related to a kind of probiotics and preparation method thereof.
Background technology
The development of antibiotic use to animal husbandry serves great impetus, along with science and growth in the living standard, people recognize gradually, antibiotic makes an addition in feed for a long time, animal pathogen is developed immunity to drugs, causes antibiotic dosage to increase, cause livestock and poultry autogenous infection and suprainfection, cause the bad consequence such as imbalance of the decline of animal body immunity, animal gastrointestinal tract flora, also worked the mischief by the health of food chain to the mankind.At present, the U.S. and some countries of West Europe in succession forbid or limit feeding antibiotic, and therefore, developing antibiotic substitute has become in animal farming industry one problem anxious to be resolved.
Probiotics as a kind of nuisanceless, noresidue green feed additive and there is the growth promoting animal, improve the immunity of animal, reduce animal productiong becomes investigation and application focus to advantages such as the pollution levels of environment, the research of China's probiotics starts from the seventies in last century, and the later stage eighties just starts the research and development paying attention to additive for microbe feedstuff.Because cyclodextrin has hydrophobic outer hydrophilic structure in uniqueness, the guest molecule suitable with many sizes forms inclusion compound, can form liquid inclusion complex by cyclodextrin to Ceftiofur inclusion.The formation of inclusion compound can avoid the reactions such as oxidation or hydrolysis by available protecting compound, and also increase the water-soluble of medicine simultaneously, strengthen stability, reduce toxic and side effect, Drug controlled release improves bioavailability.Recently, the research application of the cyclodextrin physical chemistry and biological property aspect that are used for improving medicine more and more receives the concern of people.Particularly be used for improving the stability of dewatering medicament and bioavilability, have the report of clathration in some solution, and the preparation of cyclodextrin encapsulated probiotics and property Quality Research thereof have no report.
Summary of the invention
Object of the present invention is exactly for above-mentioned defect of the prior art, provides a kind of probiotics and preparation method thereof.
To achieve these goals, technical scheme provided by the invention is: a kind of probiotics, and described probiotics is Benexate Hydrochloride.
Further, above-mentioned a kind of probiotics, described Benexate Hydrochloride is Mixed culture again after lactic acid bacteria, bacillus and saccharomycete being cultivated respectively, finally adds that beta-cyclodextrin inclusion compound obtains.
Second object of the present invention there is provided the preparation method of above-mentioned a kind of probiotics, comprises the following steps:
1) first three kinds of bacterium powder being made into quality percent by volume (W/V) is the liquid of 5%, 0.1ml lactic acid bacteria, 0.1ml bacillus and 0.1ml saccharomycete are inoculated in respectively in MRS fluid nutrient medium, meat soup fluid nutrient medium and potato dextrose broth, be placed in 37 DEG C of incubators and cultivate taking-up after 24 hours, obtain lactic acid bacteria culture solution, bacillus nutrient solution and saccharomycete nutrient solution;
2) lactic acid bacteria culture solution step 1) obtained, bacillus nutrient solution and saccharomycete nutrient solution mix with the vinasse after sterilizing, wheat bran according to the ratio of volume ratio 2:1:2, be placed in 31.8 DEG C of incubators again to cultivate after 24 hours and obtain microorganism nutrient chemical, it is 200 times amount of three kinds of bacteria culture fluid gross masses that the addition of vinasse and wheat bran is;
3) by step 2) add beta-schardinger dextrin-in the microorganism nutrient chemical that obtains and carry out inclusion, inclusion temperature is-20 DEG C-65 DEG C, mixing time is 2-5 hour, the additional proportion of beta-schardinger dextrin-is according to mass ratio range microorganism nutrient chemical: beta-schardinger dextrin-is 10:(1-3), be drying to obtain probiotics.
Further, the preparation method of above-mentioned a kind of probiotics, in described step 3), inclusion temperature is 20 DEG C, and mixing time is 4 hours.
Beneficial effect of the present invention is: a kind of probiotics provided by the invention and preparation method thereof; because beta-schardinger dextrin-has hydrophobic outer hydrophilic structure in uniqueness, the guest molecule suitable with many sizes forms inclusion compound; comparatively stable inclusion compound can be formed to Ceftiofur inclusion by beta-schardinger dextrin-; the formation of inclusion compound can avoid the reactions such as oxidation or hydrolysis by available protecting compound; also increase the stability of medicine simultaneously; strengthen water-soluble; reduce toxic and side effect, control probiotics release and improve bioavailability.
Detailed description of the invention
reagent source used in technical solution of the present invention:
Lactic acid bacteria: Cangzhou Co., Ltd of prosperous generation thing technical research institute (RS20140122-3);
Bacillus (separating shallow lake bacillus): Rui Gu bio tech ltd, Baoding (20140308YB);
Saccharomycete: Cangzhou Co., Ltd of prosperous generation thing technical research institute (XM20140220-7);
MRS fluid nutrient medium: Shanghai Lu Feng Chemical Co., Ltd. (60541609-2216e);
Meat soup fluid nutrient medium: Shanghai Lu Feng Chemical Co., Ltd. (60541609-2200a);
Potato dextrose broth: Shanghai Gu Chen Bioisystech Co., Ltd (P0185);
Beta-schardinger dextrin-: the biological Co., Ltd (20131222) of Mengzhou City of Henan Province foundation for a great undertaking.
embodiment 1:
A preparation method for probiotics, comprises the following steps:
1) first three kinds of bacterium powder are made into 5%(W/V) liquid, 0.1ml lactic acid bacteria, 0.1ml bacillus and 0.1ml saccharomycete are inoculated in respectively in MRS fluid nutrient medium, meat soup fluid nutrient medium and potato dextrose broth, be placed in 37 DEG C of incubators and cultivate taking-up after 24 hours, obtain lactic acid bacteria culture solution, bacillus nutrient solution and saccharomycete nutrient solution;
2) lactic acid bacteria culture solution step 1) obtained, bacillus nutrient solution and saccharomycete nutrient solution mix with the vinasse after sterilizing, wheat bran according to the ratio of volume ratio 2:1:2, be placed in 31.8 DEG C of incubators again to cultivate after 24 hours and obtain microorganism nutrient chemical, it is 200 times amount of three kinds of bacteria culture fluid gross masses that the addition of vinasse and wheat bran is;
3) by step 2) add beta-schardinger dextrin-in the microorganism nutrient chemical that obtains and carry out inclusion, inclusion temperature is-20 DEG C DEG C, mixing time is 2 hours, and the additional proportion of beta-schardinger dextrin-is according to mass ratio range microorganism nutrient chemical: beta-schardinger dextrin-is 10:1, is drying to obtain probiotics.
embodiment 2:
A preparation method for probiotics, comprises the following steps:
1) first three kinds of bacterium powder are made into 5%(W/V) liquid, 0.1ml lactic acid bacteria, 0.1ml bacillus and 0.1ml saccharomycete are inoculated in respectively in MRS fluid nutrient medium, meat soup fluid nutrient medium and potato dextrose broth, be placed in 37 DEG C of incubators and cultivate taking-up after 24 hours, obtain lactic acid bacteria culture solution, bacillus nutrient solution and saccharomycete nutrient solution;
2) lactic acid bacteria culture solution step 1) obtained, bacillus nutrient solution and saccharomycete nutrient solution mix with the vinasse after sterilizing, wheat bran according to the ratio of volume ratio 2:1:2, be placed in 31.8 DEG C of incubators again to cultivate after 24 hours and obtain microorganism nutrient chemical, it is 200 times amount of three kinds of bacteria culture fluid gross masses that the addition of vinasse and wheat bran is;
3) by step 2) add beta-schardinger dextrin-in the microorganism nutrient chemical that obtains and carry out inclusion, inclusion temperature is 65 DEG C, mixing time is 5 hours, and the additional proportion of beta-schardinger dextrin-is according to mass ratio range microorganism nutrient chemical: beta-schardinger dextrin-is 10:3, is drying to obtain probiotics.
embodiment 3:
A preparation method for probiotics, comprises the following steps:
1) first three kinds of bacterium powder are made into 5%(W/V) liquid, 0.1ml lactic acid bacteria, 0.1ml bacillus and 0.1ml saccharomycete are inoculated in respectively in MRS fluid nutrient medium, meat soup fluid nutrient medium and potato dextrose broth, be placed in 37 DEG C of incubators and cultivate taking-up after 24 hours, obtain lactic acid bacteria culture solution, bacillus nutrient solution and saccharomycete nutrient solution;
2) lactic acid bacteria culture solution step 1) obtained, bacillus nutrient solution and saccharomycete nutrient solution mix with the vinasse after sterilizing, wheat bran according to the ratio of volume ratio 2:1:2, be placed in 31.8 DEG C of incubators again to cultivate after 24 hours and obtain microorganism nutrient chemical, it is 200 times amount of three kinds of bacteria culture fluid gross masses that the addition of vinasse and wheat bran is;
3) by step 2) add beta-schardinger dextrin-in the microorganism nutrient chemical that obtains and carry out inclusion, inclusion temperature is 20 DEG C, mixing time is 4 hours, and the additional proportion of beta-schardinger dextrin-is according to mass ratio range microorganism nutrient chemical: beta-schardinger dextrin-is 10:2, is drying to obtain probiotics.
embodiment 4:
One, the mensuration of number of viable in probiotics:
1, material prepare: first three kinds of bacterium powder are made into 5%(W/V) liquid, with micro liquid sampler draw 10
-6-10
-8times liquid each 0.1ml coating is inoculated in (each dilution factor establishes two repetition) in MRS solid medium plate, is placed in after 24h cultivated by 37 DEG C of incubators and takes out, carry out lactic acid bacteria colony counting, with micro liquid sampler absorption 10
-6-10
-8times liquid each 0.1ml coating is inoculated in (each dilution factor establishes two repetition) in potato glucose solid medium plate, is placed in after 24h cultivated by 37 DEG C of incubators and takes out, carry out saccharomycete colony counting, with micro liquid sampler absorption 10
-6-10
-8times liquid each 0.lml coating is inoculated in (each dilution factor establishes two repetitions) in meat soup solid medium plate, is placed in after 24h cultivated by 37 DEG C of incubators and takes out, carry out bacillus bacterium colony counting.
2, the drying of probiotics: first record probiotics in freeze drying by pilot study, shady and cool place is air-dry, 25 DEG C of oven for drying, 45 DEG C of oven for drying, the time that after 65 DEG C of oven for drying dryings, moisture is used between 12%-13% is respectively 8h, 12h, 10h, 8h and 6h, probiotics is divided into five processed group, often organize three repetitions, each repetition 500g adopts freeze drying respectively, shady and cool place is air-dry, 25 DEG C of oven dry, dry and 65 DEG C of oven dry, five kinds of different drying modes for 45 DEG C, drying is carried out to the probiotics of five groups, detect the lactic acid bacteria in dry latter each group, the content of saccharomycete and bacillus.
3, result:
Different drying mode is on the impact (1g of number of viable in probiotics
cFU/g), result is as shown in table 1.
Table 1
。
As can be seen from Table 1, in freeze drying group, air-dry group, 25 DEG C oven dry groups, 45 DEG C of oven dry groups, 65 DEG C of oven dry groups, significantly (P<0.01), contained by freeze drying group, lactic acid bacterium number is maximum for contained lactic acid bacterium number and inclusion and non-inclusion inequality heteropole.Along with the rising of bake out temperature, the quantity of lactic acid bacteria extremely significantly reduces, and by being higher than the preparation of not inclusion after inclusion to the stability of heat.Contained saccharomycete quantity and inclusion and non-inclusion inequality heteropole remarkable (P <0.01) in freeze drying group, air-dry group, 25 DEG C oven dry groups, 45 DEG C of oven dry groups, 65 DEG C of oven dry groups, in freeze drying group, contained saccharomycete quantity is maximum, along with the rising of bake out temperature, saccharomycetic quantity extremely significantly reduces (P <0.01), and by being higher than the preparation of not inclusion after inclusion to the stability of heat.In freeze drying group, air-dry group, 25 DEG C oven dry groups, 45 DEG C of oven dry groups, 65 DEG C of oven dry groups, inclusion compound preparation is not containing bud pole bacterium quantity variance significantly (P>0.05), bud pole bacterial content in wherein freeze drying group is the highest, but along with the rising of bake out temperature, the quantity of bud pole bacterium presents the trend reduced gradually.In non-inclusion group freeze drying group, air-dry group, 25 DEG C oven dry groups, 45 DEG C of oven dry groups, 65 DEG C of oven dry groups, contained saccharomycete quantity and difference are extremely significantly (P <0.01).
Two, the assay method of fodder-grade micro-ecological preparation granulating process and viable bacteria content:
1, the composition of feed in granulating process:
Feed is grower pigs powder, and its formula is 22% dregs of beans, 54% corn, 40% premix, 200kg oil chaff.
2, granulating process:
According to the addition of 1g/kg, probiotics after inclusion is added in grower pigs mash feed, in mixer, mixes 1Omin, these mash feeds mixed with the probiotics after inclusion are divided into five groups, often organize three repetitions, each weight 5kg.The group of four is wherein granulated, in granulating process process, selects 15 seconds, 30 seconds, 45 seconds, 60 seconds conditioning period to carry out modifier treatment respectively, produce the particulate material of four groups of different conditioning period.
3, the mensuration of viable bacteria content in feed granulating:
According to the particulate material producing four groups of different conditioning period, adopt the method identical with micro-flora content in detection probiotics to count the viable bacteria in mash feed and four groups of particulate material after sampling, different conditioning period is on the impact (lg of number of viable in probiotics
cFU/g), result is as shown in table 2.
Table 2
。
As can be seen from Table 2, in preparation 0 second, 15 seconds, 30 seconds, 45 seconds, the 60 seconds conditioning period groups of non-inclusion, contained lactic acid bacterium number difference extremely significantly (P <0.01), along with the increase of conditioning period, the quantity of lactic acid bacteria extremely significantly reduces (P <0.01), and the difference of inclusion is not remarkable.Contained saccharomycete in preparation 0 second, 15 seconds, 30 seconds, 45 seconds, the 60 seconds conditioning period groups of non-inclusion, bud pole bacterium quantity variance are remarkable.And the difference of inclusion is not remarkable.In the present invention; under cyclodextrin safeguard measure is taked to said preparation; after probiotics is granulated under 60 seconds conditioning period by the mash feed of the addition preparation of 0.1%; the logarithm value of contained lactic acid bacteria, saccharomycete, bacillus quantity is respectively 7.04,7.15,7.46 compared with the protozoa preparation not carrying out granulating, and three kinds of bacterium quantity all do not reduce significantly.Test proves, non-inclusion probiotics is not suitable for granulating under these conditions, but by betadex, can prepare particle by this technique.
Three, probiotics is on the impact of microorganism in grower pigs production performance and excrement:
1, test material and method:
Fodder-grade micro-ecological preparation is granulated: self-control (see test one).
2, experimental animal grouping and Diet Formula "
Test the DLY three way cross grower pigs 150 selecting about body weight 25kg, health status basically identical.Be divided into five processed group at random, i.e. 1 blank group, 1 control group, 3 test group, each process 3 repetition, each repetition 10 pigs, male and female half and half.Blank group is fed basal diet; The daily ration (l00ml/kg Kitasamycin and 300mg/mg terramycin) of control group fed basal diet added with antibiotic; Test group I basal diet of feeding adds the daily ration of 500mg/kg probiotics (in daily ration, content is 0.05%); Test group II basal diet of feeding adds the daily ration of 1000m/kg probiotics (in daily ration, content is 0.10%); Test group III basal diet of feeding adds the daily ration of 1500mg/kg probiotics (in daily ration, content is 0.15%).Concrete experimental design is as shown in table 3 with grouping.
Table 3
。
3, results and analysis:
Probiotics is on the impact of grower pigs production performance, and result is as shown in table 4.
Table 4
。
As can be seen from Table 4, there is not significant difference in each group feed intake, the average daily gain of test group I test group II test group III, compared with blank group, improves 7.14%, 9.04%, 7.62% respectively; Close with control group.Compared with blank group, the diarrhea rate of test group I test group II test group III reduces, close with control group.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (4)
1. a probiotics, is characterized in that, described probiotics is Benexate Hydrochloride.
2. a kind of probiotics according to claim 1, is characterized in that, described Benexate Hydrochloride is Mixed culture again after lactic acid bacteria, bacillus and saccharomycete being cultivated respectively, finally adds that beta-cyclodextrin inclusion compound obtains.
3. the preparation method of a kind of probiotics as claimed in claim 1 or 2, is characterized in that, comprise the following steps:
1) three kinds of bacterium powder are made into the liquid that quality percent by volume is 5%, again 0.1ml lactic acid bacteria, 0.1ml bacillus and 0.1ml saccharomycete are inoculated in MRS fluid nutrient medium, meat soup fluid nutrient medium and potato dextrose broth respectively, be placed in 37 DEG C of incubators and cultivate taking-up after 24 hours, obtain lactic acid bacteria culture solution, bacillus nutrient solution and saccharomycete nutrient solution;
2) lactic acid bacteria culture solution step 1) obtained, bacillus nutrient solution and saccharomycete nutrient solution mix with the vinasse after sterilizing, wheat bran according to the ratio of volume ratio 2:1:2, be placed in 31.8 DEG C of incubators again to cultivate after 24 hours and obtain microorganism nutrient chemical, it is 200 times amount of three kinds of bacteria culture fluid gross masses that the addition of vinasse and wheat bran is;
3) by step 2) add beta-schardinger dextrin-in the microorganism nutrient chemical that obtains and carry out inclusion, inclusion temperature is-20 DEG C-65 DEG C, mixing time is 2-5 hour, the additional proportion of beta-schardinger dextrin-is according to mass ratio range microorganism nutrient chemical: beta-schardinger dextrin-is 10:(1-3), be drying to obtain probiotics.
4. the preparation method of a kind of probiotics according to claim 3, is characterized in that, in described step 3), inclusion temperature is 20 DEG C, and mixing time is 4 hours.
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| CN105412156A (en) * | 2015-11-11 | 2016-03-23 | 郑州后羿制药有限公司 | Preparation method of microecological preparation for poultry |
| CN106561976A (en) * | 2016-11-07 | 2017-04-19 | 广西来宾八百里农业有限公司 | Preparing method of microecological preparation |
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Application publication date: 20150408 |