CN105154345A - Preparation method of composite probiotic preparation added into ruminant feed - Google Patents
Preparation method of composite probiotic preparation added into ruminant feed Download PDFInfo
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Abstract
The invention discloses a preparation method of a composite probiotic preparation added into ruminant feed, belonging to the technical field of animal feed processing. The preparation method comprises the following steps: carrying out activation culture on Candida krusei, Candida utilis and Streptococcus thermophilus used as strains, carrying out strain fermentation by using glucose and the like as raw materials and adding sodium selenite to convert the sodium selenite into organic selenium, wherein glycerol and other substances are generated in the fermentation process; and finally, adding corn flour as an adsorption carrier, and concentrating to obtain the composite probiotic preparation added into ruminant feed. The example proves that the probiotic prepared by the method has the advantages of high combination rate with rumination feed and no toxicity or harm, contains abundant glycerol and organic selenium, and enhances the absorptivity of the ruminant.
Description
Technical field
The invention discloses a kind of preparation method adding composite probiotics preparations in ruminant feed to, belong to animal-feed processing technique field.
Background technology
Probiotic bacterium is the living microorganism that a class is useful to host, is to be colonizated in human intestinal, reproductive system, can produce definite health efficacy thus improve host's microecological balance, plays the active beneficial microorganism general name of beneficial effect.Bacterium useful in human body, animal body or fungi mainly contain: clostridium butyricum, milk-acid bacteria, bifidus bacillus, Lactobacterium acidophilum, actinomycetes, yeast etc.The composite reactive probiotic bacterium of the most powerful product of the function studied in the world at present mainly above each quasi-microorganism composition, it is widely used in biotechnology, industrial or agricultural, food safety and life and health field.
Probiotic bacterium is very high with using value in the closely-related key areas of human lives at industry, agricultural, medicine, food, feed etc., improving micro-ecological environment in animal body, increases immunizing power, preventing disease, growth promoting effects and improves in breeding environment etc. and produce huge economic benefits and social effect.
The ability that ruminating animal directly absorbs glucose is very limited, and dam experiences a series of violent physiological change such as gestation, childbirth, beginning lactation in perinatal period, very easily causes energy metabolism impairment.Glycerine can add in the daily ration of ruminating animal as anti-insulin principle, can regulate plasma metabolism, improves milk performance, increases blood sugar, reduces the content of BHBA and NEFA in blood, reduces the ketoboidies content in urine, and then adjusting energy balance.Selenium is the necessary trace element of a kind of animal, and its topmost function is antioxygenation.Perinatal period, the generation of ruminating animal activity in vivo oxygen greatly exceed normal level, and selenium can participate in the Antioxidative Defense System of body.Widely used benefit selenium method adds Sodium Selenite in daily ration then, but it is lower but to there is combination rate, and toxicity is large, pollutes large, therefore and by limitation use.
Summary of the invention
The technical problem that the present invention mainly solves: the ability directly absorbing glucose for current ruminating animal self is very limited, and the selenium necessary trace element that is animal, but mending selenium method is at present add Sodium Selenite in daily ration, cause combination rate low, toxicity is large, with serious pollution problem, provide a kind of preparation method adding composite probiotics preparations in ruminant feed to, the method is with candida krusei bacterium, product Ruan's candidiasis and thermophilus streptococcus are that bacterial classification carries out activation culture, with glucose etc. for raw material, add Sodium Selenite, organoselenium is translated into by strain fermentation, incidentally the materials such as glycerine are produced in the process of fermentation, finally add Semen Maydis powder as absorption carrier, namely obtain after concentrated and a kind ofly add composite probiotics preparations in ruminant feed to.Probiotic bacterium obtained by the present invention not only with to ruminate feed combination rate high, nontoxic, and is rich in a large amount of glycerine and organoselenium, and ruminating animal receptivity is strengthened.
In order to solve the problems of the technologies described above, the technical solution adopted in the present invention is:
(1) the candida krusei strain at 4 ~ 6 DEG C will be kept at, be inoculated in solid activation medium, 24 ~ 30h is cultivated at 30 ~ 40 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 24 ~ 30h with 100 ~ 150r/min at 30 ~ 35 DEG C obtains seed liquor 1;
(2) strain of Ruan's candiyeast will be produced, be inoculated in solid activation medium, 30 ~ 36h is cultivated at 28 ~ 30 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 28 ~ 38 DEG C, constant temperature culture 18 ~ 24h obtains seed liquor 2;
(3) by strains of streptococcus thermophilus, be inoculated in solid activation medium, 36 ~ 48h is cultivated at 38 ~ 42 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 35 ~ 40 DEG C, constant temperature culture 24 ~ 30h obtains seed liquor 3;
(4) in fermentor tank, add 400 ~ 500g glucose, 300 ~ 400g yeast extract, 60 ~ 70g peptone, 30 ~ 40g ammonium sulfate and 30 ~ 40g hydrogen sulfate dipotassium, add 20 ~ 30L distilled water again, 10 ~ 15min is stirred at the rotating speed of 100 ~ 200r/min with magnetic stirrer, high pressure steam sterilization 5 ~ 10min at 100 ~ 115 DEG C, is cooled to room temperature afterwards;
(5) in above-mentioned cooled fermentor tank, sodium selenite solution, 100 ~ 150mL seed liquor 1,200 ~ 300mL seed liquor 2 and the 700 ~ 800mL seed liquor 3 that 400 ~ 500mL selenium content is 0.5 ~ 0.6mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 100 ~ 200r/min stirring 15 ~ 20min again, be decompressed to 40 ~ 50kPa in tank, at 30 ~ 40 DEG C, obtain fermented liquid after ferment at constant temperature 30 ~ 36h;
(6) get the above-mentioned fermented liquid of 300 ~ 400mL and put into 1000mL beaker, add 200 ~ 300g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, 900 ~ 1000Pa is decompressed in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
Described solid activation medium is made up of 20 ~ 30g glucose, 2 ~ 4g urea, 4 ~ 5g yeast extract, 20 ~ 30g agar and 900 ~ 1000mL distilled water.
Described seed culture medium is made up of 200 ~ 250g glucose, 2 ~ 5g urea, 4 ~ 6g yeast extract, 20 ~ 30g agar and 1000 ~ 1200mL distilled water.
Described candida krusei strain code name is CICC1722, and thermophilus streptococcus code name is AS1.2471, and producing Ruan's candida bacterial strain code name is AS2.1180.
Application method of the present invention is: the ruminant feed compound probiotic concentrated solution obtained by the present invention is added water and dilutes 100 times, then in every 100 ~ 300g ruminant feed, add the probiotic bacterium liquid after 5 ~ 15mL dilution, stir, put into animal foodstuff basin, 10 ~ 20 days, obviously can feel that ruminating animal immunizing power improves, body weight growth is rapidly, vivaciously healthy.
The invention has the beneficial effects as follows:
(1) probiotic bacterium obtained by the present invention is added in the edible feed of ruminating animal, be rich in glycerine and organoselenium, ruminating animal receptivity is strengthened;
(2) probiotic bacterium is combined with ruminant feed, causes combination rate to improve, and without any toxicity, for environment without any pollution problem.
Embodiment
First the candida krusei strain at 4 ~ 6 DEG C will be kept at, be inoculated in solid activation medium, 24 ~ 30h is cultivated at 30 ~ 40 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 24 ~ 30h with 100 ~ 150r/min at 30 ~ 35 DEG C obtains seed liquor 1; Then the strain of Ruan's candiyeast will be produced, be inoculated in solid activation medium, 30 ~ 36h is cultivated at 28 ~ 30 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 28 ~ 38 DEG C, constant temperature culture 18 ~ 24h obtains seed liquor 2; Subsequently by strains of streptococcus thermophilus, be inoculated in solid activation medium, 36 ~ 48h is cultivated at 38 ~ 42 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 35 ~ 40 DEG C, constant temperature culture 24 ~ 30h obtains seed liquor 3; Backward fermentor tank in add 400 ~ 500g glucose, 300 ~ 400g yeast extract, 60 ~ 70g peptone, 30 ~ 40g ammonium sulfate and 30 ~ 40g hydrogen sulfate dipotassium, add 20 ~ 30L distilled water again, 10 ~ 15min is stirred at the rotating speed of 100 ~ 200r/min with magnetic stirrer, high pressure steam sterilization 5 ~ 10min at 100 ~ 115 DEG C, is cooled to room temperature afterwards; Next in above-mentioned cooled fermentor tank, sodium selenite solution, 100 ~ 150mL seed liquor 1,200 ~ 300mL seed liquor 2 and the 700 ~ 800mL seed liquor 3 that 400 ~ 500mL selenium content is 0.5 ~ 0.6mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 100 ~ 200r/min stirring 15 ~ 20min again, be decompressed to 40 ~ 50kPa in tank, at 30 ~ 40 DEG C, obtain fermented liquid after ferment at constant temperature 30 ~ 36h;
Finally get the above-mentioned fermented liquid of 300 ~ 400mL and put into 1000mL beaker, add 200 ~ 300g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, 900 ~ 1000Pa is decompressed in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
Described solid activation medium is made up of 20 ~ 30g glucose, 2 ~ 4g urea, 4 ~ 5g yeast extract, 20 ~ 30g agar and 900 ~ 1000mL distilled water.
Described seed culture medium is made up of 200 ~ 250g glucose, 2 ~ 5g urea, 4 ~ 6g yeast extract, 20 ~ 30g agar and 1000 ~ 1200mL distilled water.
Described candida krusei strain code name is CICC1722, and thermophilus streptococcus code name is AS1.2471, and producing Ruan's candida bacterial strain code name is AS2.1180.
Example 1
First the candida krusei strain at 4 DEG C will be kept at, be inoculated in solid activation medium, 24h is cultivated at 30 DEG C, repeat switching afterwards 3 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 24h with 100r/min at 30 DEG C obtains seed liquor 1; Then will produce the strain of Ruan's candiyeast, and be inoculated in solid activation medium, at 28 DEG C, cultivate 30h, repeat switching afterwards 3 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 28 DEG C, constant temperature culture 18h obtains seed liquor 2; Subsequently by strains of streptococcus thermophilus, be inoculated in solid activation medium, at 38 DEG C, cultivate 36h, repeat switching afterwards 3 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 35 DEG C, constant temperature culture 24h obtains seed liquor 3; Backward fermentor tank in add 400g glucose, 300g yeast extract, 60g peptone, 30g ammonium sulfate and 30g hydrogen sulfate dipotassium, add 20L distilled water again, stir 10min with magnetic stirrer at the rotating speed of 100r/min, high pressure steam sterilization 5min at 100 DEG C, is cooled to room temperature afterwards; Next in above-mentioned cooled fermentor tank, sodium selenite solution, 100mL seed liquor 1,200mL seed liquor 2 and the 700mL seed liquor 3 that 400mL selenium content is 0.5mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 100r/min stirring 15min again, be decompressed to 40kPa in tank, at 30 DEG C, obtain fermented liquid after ferment at constant temperature 30h; Finally get the above-mentioned fermented liquid of 300mL and put into 1000mL beaker, add 200g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, be decompressed to 900Pa in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
This case method is unique novel, during use, the ruminant feed compound probiotic concentrated solution obtained by the present invention is added water and dilute 100 times, then in every 100g ruminant feed, add the probiotic bacterium liquid after 5mL dilution, stir, put into animal foodstuff basin, obviously can feel after 10 days that ruminating animal immunizing power improves, body weight growth is rapidly, vivaciously healthy.
Example 2
First the candida krusei strain at 5 DEG C will be kept at, be inoculated in solid activation medium, 26h is cultivated at 35 DEG C, repeat switching afterwards 4 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 26h with 130r/min at 33 DEG C obtains seed liquor 1; Then will produce the strain of Ruan's candiyeast, and be inoculated in solid activation medium, at 29 DEG C, cultivate 33h, repeat switching afterwards 4 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 35 DEG C, constant temperature culture 22h obtains seed liquor 2; Subsequently by strains of streptococcus thermophilus, be inoculated in solid activation medium, at 40 DEG C, cultivate 42h, repeat switching afterwards 4 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 38 DEG C, constant temperature culture 26h obtains seed liquor 3; Backward fermentor tank in add 450g glucose, 350g yeast extract, 65g peptone, 35g ammonium sulfate and 35g hydrogen sulfate dipotassium, add 25L distilled water again, stir 13min with magnetic stirrer at the rotating speed of 150r/min, high pressure steam sterilization 7min at 105 DEG C, is cooled to room temperature afterwards; Next in above-mentioned cooled fermentor tank, sodium selenite solution, 130mL seed liquor 1,250mL seed liquor 2 and the 750mL seed liquor 3 that 450mL selenium content is 0.55mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 150r/min stirring 18min again, be decompressed to 45kPa in tank, at 35 DEG C, obtain fermented liquid after ferment at constant temperature 33h; Finally get the above-mentioned fermented liquid of 350mL and put into 1000mL beaker, add 250g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, be decompressed to 950Pa in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
This case method is unique novel, during use, the ruminant feed compound probiotic concentrated solution obtained by the present invention is added water and dilute 80 times, then in every 200g ruminant feed, add the probiotic bacterium liquid after 10mL dilution, stir, put into animal foodstuff basin, obviously can feel after 15 days that ruminating animal immunizing power improves, body weight growth is rapidly, vivaciously healthy.
Example 3
First the candida krusei strain at 6 DEG C will be kept at, be inoculated in solid activation medium, 30h is cultivated at 40 DEG C, repeat switching afterwards 5 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 30h with 150r/min at 35 DEG C obtains seed liquor 1; Then will produce the strain of Ruan's candiyeast, and be inoculated in solid activation medium, at 30 DEG C, cultivate 36h, repeat switching afterwards 5 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 38 DEG C, constant temperature culture 24h obtains seed liquor 2; Subsequently by strains of streptococcus thermophilus, be inoculated in solid activation medium, at 42 DEG C, cultivate 48h, repeat switching afterwards 5 times, so that bacterial strain vigor is restored, after having activated, to produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 40 DEG C, constant temperature culture 30h obtains seed liquor 3; Backward fermentor tank in add 500g glucose, 400g yeast extract, 70g peptone, 40g ammonium sulfate and 40g hydrogen sulfate dipotassium, add 30L distilled water again, stir 15min with magnetic stirrer at the rotating speed of 200r/min, high pressure steam sterilization 10min at 115 DEG C, is cooled to room temperature afterwards; Next in above-mentioned cooled fermentor tank, sodium selenite solution, 150mL seed liquor 1,300mL seed liquor 2 and the 800mL seed liquor 3 that 500mL selenium content is 0.6mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 200r/min stirring 20min again, be decompressed to 50kPa in tank, at 40 DEG C, obtain fermented liquid after ferment at constant temperature 36h; Finally get the above-mentioned fermented liquid of 400mL and put into 1000mL beaker, add 300g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, be decompressed to 1000Pa in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
This case method is unique novel, during use, the ruminant feed compound probiotic concentrated solution obtained by the present invention is added water and dilute 60 times, then in every 300g ruminant feed, add the probiotic bacterium liquid after 15mL dilution, stir, put into animal foodstuff basin, obviously can feel after 20 days that ruminating animal immunizing power improves, body weight growth is rapidly, vivaciously healthy.
Claims (4)
1. add a preparation method for composite probiotics preparations in ruminant feed to, it is characterized in that concrete preparation process is:
(1) the candida krusei strain at 4 ~ 6 DEG C will be kept at, be inoculated in solid activation medium, 24 ~ 30h is cultivated at 30 ~ 40 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, by candida krusei bacterium inclined plane inoculating in seed culture medium, put into constant-temperature table incubator, the rotating speed constant temperature culture 24 ~ 30h with 100 ~ 150r/min at 30 ~ 35 DEG C obtains seed liquor 1;
(2) strain of Ruan's candiyeast will be produced, be inoculated in solid activation medium, 30 ~ 36h is cultivated at 28 ~ 30 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 28 ~ 38 DEG C, constant temperature culture 18 ~ 24h obtains seed liquor 2;
(3) by strains of streptococcus thermophilus, be inoculated in solid activation medium, 36 ~ 48h is cultivated at 38 ~ 42 DEG C, repeat switching afterwards 3 ~ 5 times, so that bacterial strain vigor is restored, after having activated, will produce Ruan's candidiasis inclined plane inoculating in seed culture medium, at 35 ~ 40 DEG C, constant temperature culture 24 ~ 30h obtains seed liquor 3;
(4) in fermentor tank, add 400 ~ 500g glucose, 300 ~ 400g yeast extract, 60 ~ 70g peptone, 30 ~ 40g ammonium sulfate and 30 ~ 40g hydrogen sulfate dipotassium, add 20 ~ 30L distilled water again, 10 ~ 15min is stirred at the rotating speed of 100 ~ 200r/min with magnetic stirrer, high pressure steam sterilization 5 ~ 10min at 100 ~ 115 DEG C, is cooled to room temperature afterwards;
(5) in above-mentioned cooled fermentor tank, sodium selenite solution, 100 ~ 150mL seed liquor 1,200 ~ 300mL seed liquor 2 and the 700 ~ 800mL seed liquor 3 that 400 ~ 500mL selenium content is 0.5 ~ 0.6mg/mL is added, use magnetic stirrer with sealed can mouth after the rotating speed of 100 ~ 200r/min stirring 15 ~ 20min again, be decompressed to 40 ~ 50kPa in tank, at 30 ~ 40 DEG C, obtain fermented liquid after ferment at constant temperature 30 ~ 36h;
(6) get the above-mentioned fermented liquid of 300 ~ 400mL and put into 1000mL beaker, add 200 ~ 300g Semen Maydis powder, be placed on shaken well on ultrasonator, put into concentration tank again, 900 ~ 1000Pa is decompressed in tank, carry out concentration, the concentrated solution obtained and one add composite probiotics preparations in ruminant feed to.
2. a kind of preparation method adding composite probiotics preparations in ruminant feed to according to claim 1, is characterized in that: described solid activation medium is made up of 20 ~ 30g glucose, 2 ~ 4g urea, 4 ~ 5g yeast extract, 20 ~ 30g agar and 900 ~ 1000mL distilled water.
3. a kind of preparation method adding composite probiotics preparations in ruminant feed to according to claim 1, is characterized in that: described seed culture medium is made up of 200 ~ 250g glucose, 2 ~ 5g urea, 4 ~ 6g yeast extract, 20 ~ 30g agar and 1000 ~ 1200mL distilled water.
4. a kind of preparation method adding composite probiotics preparations in ruminant feed to according to claim 1, it is characterized in that: described candida krusei strain code name is CICC1722, thermophilus streptococcus code name is AS1.2471, and producing Ruan's candida bacterial strain code name is AS2.1180.
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Cited By (2)
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| CN109852661A (en) * | 2019-02-23 | 2019-06-07 | 湖北和诺生物工程股份有限公司 | A method of organic selenium extractant is prepared using microorganism |
| CN114854611A (en) * | 2022-02-16 | 2022-08-05 | 中国农业科学院油料作物研究所 | Selenium-enriched yeast and probiotic compound bacterium liquid and fermented soybean meal feed |
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Application publication date: 20151216 |