CN101658243A - Microbiological feed additive for adjusting energy metabolism impairment of flocks and herds during perinatal period - Google Patents
Microbiological feed additive for adjusting energy metabolism impairment of flocks and herds during perinatal period Download PDFInfo
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Abstract
The invention relates to microbiological feed additive for adjusting energy metabolism impairment of flocks and herds during perinatal period, which belongs to the production field of feed additives.A preparation method of the microbiological feed additive comprises the following steps: adding inorganic selenium source into a microbial inoculum culture medium to ensure that the concentration of selenium in the culture medium is 2 to 10 mg/kg; inoculating Candida krusei, Candida valida and Streptococcus thermophilus seed liquor in the culture medium according to the ratio of 13:1:2; mixing theculture solution with the a proper amount of corn flour, and drying at 35 to 40 DEG C to obtain the feed additive. The invention adopts the principle that probiotics can convert sugar into glycerin and convert inorganic selenium into organic selenium to produce the complex probiotic product which is rich in glycerin and organic selenium, can exert the efficacies of glycerin, organic selenium andprobiotics simultaneously, can supply energy to ruminant, regulate and control the energy metabolism, adjust intestinal flora, resist pathogenic microorganism, increase appetite, enhance body immunityand disease resistance, improve the production performance, and is a safe and novel feed additive with has high value and no residual.
Description
Technical field
The present invention relates to a kind of additive for microbe feedstuff that is used for adjusting energy metabolism impairment of flocks and herds during perinatal period, belong to the feed addictive production field.
Background technology
Be meant that latter half of gestation (antenatal 2～3 weeks) is to the lactation initial stage (2～3 weeks of postpartum) perinatal period of ox, sheep.Because most of sugar forms volatile fatty acid in the ruminant in the lumen fermentation process, have only a small amount of glucose in alimentary canal, to be absorbed, and perinatal period, dam experienced a series of violent physiological change such as gestation, childbirth, very easily cause energy insufficiency of intake and consumption to increase, blood sugar concentration descends, thereby causes energy metabolism impairment.As milk cow milk fever, ketoacidosis and fatty liver take place, eclampsia, ketoacidosis etc. take place in milch goat.With the ketosis of dairy cows is example, and document announcement is arranged, and the incidence of disease of China's ketosis of dairy cows accounts for 15.0%～30.0% of cow in milk, and the U.S. is 5.0%, and Britain is 12.0%, and India is 14.69%, and Japan is up to 43.1%.Therefore, perinatal period, energy metabolism impairment had a strong impact on ox, sheep aquaculture.
Gluconeogenesis is meant that nonsugars such as glycerine, lactic acid and some amino acid change the process of glycogen or glucose into.Perinatal gluconeogenesis has important meaning to cattle and sheep, is the important channel of regulating energy metabolism impairment.As far back as the seventies in 20th century just the someone propose glycerine and can be used as anti-insulin principle and add in the daily ration of ruminant, current searching new forms of energy press for the energy that also makes glycerine become a kind of practicality.Discover, can improve blood sugar for the perinatal period milk cow glycerine of feeding, regulate the blood metabolism, improve milk performance, reduce ketoboidies content, regulate energy balance.The production method of glycerine mainly contains three kinds: the one, from the byproduct of natural oil saponification or hydrolysis, obtain, i.e. the fat saponification Hydrolyze method; The 2nd, be that raw material is crossed aldehydic acid method synthetic glycerine, i.e. chemical synthesis by epoxychloropropane method, methacrylaldehyde method, expoxy propane with petrochemicals; The 3rd, be that raw material is through microbial action fermenting and producing glycerine, i.e. fermentation method with starch, carbohydrate or agricultural byproducts.In recent years, preceding two kinds of methods are because the grease rise in price, the minimizing that soap is produced, and the cost height of chemical synthesis, reasons such as glycerol extraction complexity cause the glycerine demand under-supply, and price rises.The toxicity of industry glycerol is bigger in addition, contains arsenic and heavy metal substance, causes glycerine not to be widely used in the feed additive for ruminant field.And fermentation method is produced the shortcoming that glycerine has exactly overcome industrial production glycerine.In recent years, along with biotechnology, microorganism fungus kind, zymotechnique and each side Progress in technique such as extraction separates, fermentation glycerol production has good development prospect.Choose specific probio as bacterial classification, with starch, carbohydrate or agricultural byproducts is that raw material carries out fermenting and producing glycerine, be used for the feed additive for ruminant field then, its advantage is: raw material sources are abundant, equipment requires simple, safety non-pollution need not extract glycerine after the fermentation, greatly reduces production cost.At present, there is not fermentation method to produce the report that glycerine is directly used in feed additive field as yet.
Selenium is the necessary trace element of a kind of animal, is a kind of multi-functional life nutrient.Studies show that selenium has anti-oxidant to animal, improve immunity of organisms, improve resistance against diseases, regulate organism metabolism, improve reproductive capability, antitumor, multiple functions such as the poisonous element of antagonism.Selenium generally enters in the animal body by diet, and micro-selenium deficiency or unreasonable structure can cause that all body lacks selenium, obtain from soil and the selenium in China's feed is many, so Se content is closely related in body selenium situation and the soil in the feed.According to statistics, the whole world has more than 40 countries and regions to lack selenium, and China is scarce selenium big country, has 72% area to exist selenium deficiency.Perinatal milk cow is in stress situation, and immunosupress appears in body, and the neurological susceptibility of disease increases.Research thinks that the incidental mammitis of cow in perinatal period, hysteritis, retained afterbirth, ketoacidosis, fatty liver etc. are all relevant with the oxidative stress due to body latter half of gestation and the childbirth stimulation etc.Sordillo report, milk cow third trimester of pregnancy, childbirth and the early stage metabolism of lactation need all can make activity in vivo oxygen, and (reactiveoxygenspecies, generation ROS) increases, and has substantially exceeded normal level, and has made animal be in oxidative stress status.Under the normal condition, ROS constantly produces in the body, is also constantly removed by antioxidant system, thereby remains a kind of dynamic balance state.ROS has strong oxidizing property to many biomolecule, except that can causing protein, DNA and nucleotides damage, mainly causes biomembranous lipid peroxidation.The topmost function of selenium is a non-oxidizability, and it participates in the anti-oxidative defense system of body, lacks selenium perinatal period and can reduce body's immunological function.Glutathione peroxidase (GSH-Px) is first putative selenoenzyme in the mammalian body; has extremely strong antioxidation; catalytic reduction type glutathione (GSH) is gone back peroxide (POOH) harmful in the substance; comprise hydrogen peroxide (HOOH); make it to change into harmless hydroxy compounds, thereby the protection biomembrane is avoided the oxidative damage of peroxide.
In order to strengthen selenium in feed, control lacks the selenium symptom and takes place, and people have developed multiple selemium nutrition replenishers.The method of mending selenium at present mainly is to add inorganic selenium in feed, but studies show that in a large number the utilization rate of inorganic seleniums such as sodium selenite is lower, and the selenium that animal drains causes the danger of soil and environmental pollution bigger, and these have caused that people are to using the doubt of inorganic selenium; And organic selenium toxicity is little, the absorptivity height, and the bioavailability height does not have to pollute to environment in the scope that satisfies the required addition of animal growth substantially.Bio-organic is the easy and economic approach that obtains the organic nutrition element, helps widely applying in the actual production.The bio-organic of selenium mainly carries out in plant He in the microbial body.Studies show that in the humans and animals body, most selenium is to play a role with the biologically active form that contains selenoprotein and selenium enzyme, selenium replaces the position of sulphur in the amino acid of these protein and selenium enzyme, forms selenomethionine, selenium cystine, selenocystein etc.The organic selenium that obtains by bio-transformation is because of identical or close with its form that exists in the humans and animals body, so absorption rate is higher.
Probio (probtotlcs) has another name called regulator, bacteria-promoting agent, probiotics, microorganism formulation, active bacteria formulation, is meant " can be to host's healthy effective microorganism live body when taking in when a certain amount of ".Probio mainly comprises lactic acid bacteria class, Bifidobacterium class, bacillus category, photosynthetic bacteria class and yeast class.Perinatal period, dam experienced a series of violent physiological change such as gestation, childbirth, metatrophia, tend to cause beneficial bacterium quantity decline in the animal body enteron aisle, cause the enteric microorganism ecological disturbance, cause the flora transposition, bring out endogenous infection, cause body disease,, can keep the gut flora balance by the external source probiotic supplemented, improve immunity of organisms, reduce the neurological susceptibility of disease.
In sum, with specific probio is bacterial classification, with starch, carbohydrate or agricultural byproducts are raw material, is glycerine by fermentation method with feedstock conversion, and the selenium of inorganic states is transformed into the selenium of organic, with suitable adsorbents adsorb, obtain a kind of additive for microbe feedstuff that is used for adjusting energy metabolism impairment of flocks and herds during perinatal period that is rich in glycerine and organic selenium, can bring into play glycerine simultaneously, the triple function of organic selenium and probio, this obviously can improve the endogenous cycle border of body effectively, makeup energy, regulate energy metabolism impairment, strengthen the immunity of body, the opposing tumour, the antagonism pathogenic microorganism, thus reach prophylactic effect.Therefore the composite probiotics preparations that is rich in glycerine and organic selenium will be played a great role in medicine, feed additive field as safety, multi-functional feed addictive.
Summary of the invention
Technical problem
The purpose of this invention is to provide a kind of additive for microbe feedstuff that is used to regulate and control ox, sheep energy metabolism impairment perinatal period of simple, low-cost production, this product is brought into play glycerine, organic selenium and probio triple function simultaneously.
Technical scheme
Be used to regulate and control the additive for microbe feedstuff of ox, sheep energy metabolism impairment perinatal period, concrete production method may further comprise the steps:
1) bacterial classification candida krusei (Candida krusei), candida utili (Candida utilis), streptococcus thermophilus (Streptococcus Thermophilus); Be the probio bacterial classification that the Ministry of Public Health allows use.
2) microbial inoculum is produced culture medium quality volume ratio
15%～25% glucose, 0.5%～2% yeast extract, 0.1%～0.3% urea, 0.1%～0.3% Dried Corn Steep Liquor Powder, 0.3%～0.4% potassium dihydrogen phosphate, 4%～6% sodium chloride, all the other are water.
3) slant activation culture medium
Candida krusei activation medium (CM0357): 40g glucose, 2g urea, 4g yeast extract, 20g agar, 1000ml water, pH4.5,115 ℃ of high pressure steam sterilization 15min, standby.
Candida utili, streptococcus thermophilus activation medium (YPD): 20g glucose, 5g yeast extract, 10g peptone, 20g agar, 1000ml water, initial 6.7,115 ℃ of high pressure steam sterilization 15min of pH, standby.
4) do not add agar in the liquid activation medium slant activation culture medium.
5) sodium selenite solution of inorganic selenium source configuration selenium content 0.2～2mg/ml, 100～125 ℃ of high pressure steam sterilization 20～30min, standby.
6) adsorbent corn flour.
7) acquisition of the activation of bacterial classification and seed liquor
The candida krusei that is kept at 4 ℃ is inoculated on the CM0357 solid medium, cultivates 30～40h in 20～30 ℃; Switching is 3～5 times so continuously, be inoculated in the CM0357 fluid nutrient medium, and in 20～30 ℃, 150r/min, shaking table is cultivated 24～36h, gets the candida krusei seed liquor, and its cfu reaches 10
9More than/the ml; The candida utili that is kept at 4 ℃ is inoculated on the YPD solid medium, cultivate 30～40h in 20～30 ℃, switching is 3～5 times so continuously, be inoculated in the YPD fluid nutrient medium, in 20～30 ℃, 150r/min, shaking table is cultivated 24～36h, get the candida utili seed liquor, its cfu reaches 10
9More than/the ml; The streptococcus thermophilus that is kept at 4 ℃ is inoculated on the YPD solid medium, cultivates 36～48h in 36～45 ℃; Switching is 3～5 times so continuously, is inoculated in the YPD fluid nutrient medium, in 36～45 ℃, cultivates 24～36h, gets the streptococcus thermophilus seed liquor, and its cfu reaches 10
10More than/the ml.
8) produce
Get and produce culture medium batching glucose 15～25kg, yeast extract 0.1～1.0g, urea 100～300g, Dried Corn Steep Liquor Powder 100～300g, potassium dihydrogen phosphate 300～400g and sodium chloride 4～6kg, it is dissolved in the water, make cumulative volume reach 100L, join then in the 150L fermentation tank, 115～120 ℃ of high pressure steam sterilization 15min, be cooled to 40～50 ℃, inorganic selenium source joined microbial inoculum produce in the culture medium, make that selenium concentration is 2～10mg/kg in the culture medium; Candida krusei, candida utili and streptococcus thermophilus seed liquor were inoculated in microbial inoculum in 13: 1: 2 by volume again and produce in the culture medium, total inoculum concentration volume ratio is 6%～11%, and cultivation temperature is 20～40 ℃, and throughput is 0.05m
3/ h, shaking speed 70r/min, tank pressure are 0.04Mpa, incubation time is 96～132h;
9) product obtains
After cultivate finishing, nutrient solution mixes with an amount of corn flour, and 35～40 ℃ of oven dry promptly obtain a kind of compound probiotic product that is rich in glycerine and organic selenium.
Beneficial effect:
Test shows that the various dose of the present invention of feeding perinatal period is rich in the compound probiotic product of glycerine and organic selenium, and product treatment group plasma glucose concentration is higher than control group, and blood plasma free fatty acid and beta-hydroxybutyric acid concentration and urine ketone concentration are lower than control group.Along with the increase of daily ration interpolation product, middle dosage group plasma glucose concentration and H group are significantly higher than control group (P＜0.05); Middle dosage group and high dose group blood plasma free fatty acid concentration significantly are lower than control group (P＜0.05), and high dose group blood plasma beta-hydroxybutyric acid concentration significantly is lower than control group (P＜0.05); Middle dosage and high dose group urine ketone concentration significantly are lower than control group (P＜0.05).
The present invention utilizes probio sugar can be converted into glycerine, inorganic states selenium is converted into the principle of organic selenium, produces the compound probiotic product that is rich in glycerine and organic selenium, can bring into play glycerine, organic selenium and probio triple function simultaneously, can be the ruminant makeup energy, the regulation and control energy metabolism impairment is regulated gut flora, the antagonism pathogenic microorganism, improve a poor appetite, improve immunity of organisms and premunition, the raising production performance is a kind of safety, noresidue, the high novel fodder additive of tiring.
The specific embodiment
Embodiment 1
(1) bacterial classification
Candida krusei (Candida krusei): available from Chinese industrial microorganism fungus kind preservation administrative center, the bacterial strain code name is CICC1722.
Candida utili (Candida utilis): available from Institute of Microorganism, Academia Sinica microorganism fungus kind preservation center, the bacterial strain code name is AS2.1180.
Streptococcus thermophilus (Streptococcus Thermophilus): available from Institute of Microorganism, Academia Sinica microorganism fungus kind preservation center, the bacterial strain code name is AS1.2471.
(2) acquisition of actication of culture and seed liquor
Slant activation culture medium quality ratio: candida krusei activation medium (CM0357): 40g glucose, 2g urea, 4g yeast extract, 20g agar, 1000ml water, pH4.5,115 ℃ of high pressure steam sterilization 15min, standby; Candida utili, streptococcus thermophilus activation medium (YPD): 20g glucose, 5g yeast extract, 10g peptone, 20g agar, 1000ml water, pH6.7,115 ℃ of high pressure steam sterilization 15min, standby.
The candida krusei that is kept at 4 ℃ is inoculated on the CM0357 solid medium,, cultivates 30h in 30 ℃; Switching is 3～5 times so continuously, be inoculated in the CM0357 fluid nutrient medium, and in 30 ℃, 150r/min, shaking table is cultivated 24h, gets the candida krusei seed liquor, and its cfu reaches 3.24 * 10
9/ ml (strain liquid A).
The candida utili that is kept at 4 ℃ is inoculated on the YPD solid medium,, cultivates 32h in 28 ℃; Switching is 3～5 times so continuously, be inoculated in the YPD fluid nutrient medium, and in 28 ℃, 150r/min, shaking table is cultivated 24h, gets the candida utili seed liquor, and its cfu reaches 4.46 * 10
9/ ml (strain liquid B).
The streptococcus thermophilus that is kept at 4 ℃ is inoculated on the slant activation culture medium,, cultivates 48h in 42 ℃; Switching is 3～5 times so continuously, is inoculated in the liquid activation medium, in 42 ℃, cultivates 24h, gets the streptococcus thermophilus seed liquor, and its cfu reaches 8.25 * 10
10/ ml (strain liquid C).
(3) produce the culture medium batching
Culture medium batching: glucose 20kg, yeast extract 200g, urea 200g, Dried Corn Steep Liquor Powder 200g, potassium dihydrogen phosphate 350g, sodium chloride 4kg.
Prepare inorganic selenium source: preparation Se concentration is the sodium selenite solution 0.5L of 1.0mg/ml, and 121 ℃ of high pressure steam sterilization 30min are standby.
Adsorbent: 50kg corn flour
(4) produce
Above-mentioned microbial inoculum is produced the culture medium batching to be dissolved in the water, make its cumulative volume reach 100L, join then in BIOTECH-BS/JS (150L) fermentation tank, 115 ℃ of high pressure steam sterilization 15min, be cooled to 40～50 ℃, adding selenium content is the sodium selenite solution 0.5L of 1.0mg/ml, inserts strain liquid A6.5L, strain liquid B0.5L, strain liquid C1L, temperature is 33 ℃, and throughput is 0.05m
3/ h, revolution are 70r/min, and tank pressure is 0.04Mpa, and incubation time is 120h, obtains zymotic fluid.
With above-mentioned zymotic fluid and 50kg corn flour mixing, dry about 38 ℃, obtain being rich in the composite probiotics preparations of glycerine and organic selenium.
The amount of the composite probiotics preparations that is rich in glycerine and organic selenium of above-mentioned acquisition is 61.95kg, and the quantity of candida krusei, candida utili, streptococcus thermophilus reaches 7.75 * 10 respectively
8Cfu/g, 2.69 * 10
8Cfu/g, 6.45 * 10
9Cfu/g, glycerol content are 108.55g/kg, and the content of organic selenium is 6.79mg/kg, and the total content of selenium is 8.06mg/kg, and organic selenium conversion ratio is about 84.2%.
Embodiment 2
(1) bacterial classification
Candida krusei (Candida krusei): available from Chinese industrial microorganism fungus kind preservation administrative center, the bacterial strain code name is CICC1722.
Candida utili (Candida utilis): available from Institute of Microorganism, Academia Sinica microorganism fungus kind preservation center, the bacterial strain code name is AS2.1180.
Streptococcus thermophilus (Streptococcus Thermophilus): available from Institute of Microorganism, Academia Sinica microorganism fungus kind preservation center, the bacterial strain code name is AS1.2471.
(2) acquisition of actication of culture and seed liquor
Slant activation culture medium quality ratio: candida krusei activation medium (CM0357): 40g glucose, 2g urea, 4g yeast extract, 20g agar, 1000ml water, pH4.5,115 ℃ of high pressure steam sterilization 15min, standby; Candida utili, streptococcus thermophilus activation medium (YPD): 20g glucose, 5g yeast extract, 10g peptone, 20g agar, 1000ml water, pH6.7,115 ℃ of high pressure steam sterilization 15min, standby.
The candida krusei that is kept at 4 ℃ is inoculated on the CM0357 solid medium,, cultivates 30h in 30 ℃; Switching is 3～5 times so continuously, be inoculated in the CM0357 fluid nutrient medium, and in 30 ℃, 150r/min, shaking table is cultivated 24h, gets the candida krusei seed liquor, and its cfu reaches 3.24 * 10
9/ ml (strain liquid A).
The candida utili that is kept at 4 ℃ is inoculated on the YPD solid medium,, cultivates 32h in 28 ℃; Switching is 3～5 times so continuously, be inoculated in the YPD fluid nutrient medium, and in 28 ℃, 150r/min, shaking table is cultivated 24h, gets the candida utili seed liquor, and its cfu reaches 4.46 * 10
9/ ml (strain liquid B).
The streptococcus thermophilus that is kept at 4 ℃ is inoculated on the slant activation culture medium,, cultivates 48h in 42 ℃; Switching is 3～5 times so continuously, is inoculated in the liquid activation medium, in 42 ℃, cultivates 24h, gets the streptococcus thermophilus seed liquor, and its cfu reaches 8.25 * 10
10/ ml (strain liquid C).
(3) the required batching of fermenting
The culture medium batching: glucose 20kg, yeast extract 200g, urea 200g, Dried Corn Steep Liquor Powder 200g, potassium dihydrogen phosphate 350g, sodium chloride 4kg, all the other are water.
Prepare inorganic selenium source: preparation Se concentration is the sodium selenite solution 0.5L of 0.8mg/ml, and 121 ℃ of high pressure steam sterilization 30min are standby.
Adsorbent: 50kg corn flour
(4) produce
Microbial inoculum is produced the culture medium batching to be dissolved in the water, make its volume reach 100L, join then in BIOTECH-BS/JS (150L) fermentation tank, 115 ℃ of high pressure steam sterilization 15min, be cooled to 40～50 ℃, adding selenium content is the sodium selenite solution 0.5L of 0.8mg/ml, inserts strain liquid A6.5L, strain liquid B0.5L, strain liquid C1L, temperature is 33 ℃, and throughput is 0.05m
3/ h, revolution are 70r/min, and tank pressure is 0.04Mpa, and incubation time is 120h, obtains zymotic fluid.
With above-mentioned zymotic fluid and 50kg corn flour mixing, dry about 38 ℃, obtain being rich in the composite probiotics preparations of glycerine and organic selenium.
The amount of the composite probiotics preparations that is rich in glycerine and organic selenium of above-mentioned acquisition is 61.98kg, and the quantity of candida krusei, candida utili, streptococcus thermophilus reaches 7.77 * 10 respectively
8Cfu/g, 2.72 * 10
8Cfu/g, 6.53 * 10
9Cfu/g, glycerol content are 115.77g/kg, and the content of organic selenium is 5.70mg/kg, and the total content of selenium is 6.45mg/kg, and organic selenium conversion ratio is about 88.4%.
Experimental animal when having added the above-mentioned feed product that is rich in the composite probiotics preparations of glycerine and organic selenium and testing is chosen, processing and result of the test, comprising:
1. experimental animal and grouping
12 multiparity holstein cows, body condition, age, parity, the expected date of childbirth, the output of milk are close, be divided into 4 groups at random, every group 3 cow head, i.e. control group (not adding the product group), L group (low dose group is added the product 276.4g/ previous day that contains glycerine 30g approximately), M (middle dosage group, add the product 552.7g/ previous day contain glycerine 60g approximately), H organizes (high dose group, add the product 829.1g/ previous day contain glycerine 90g approximately), trial test 7 days, formal test from antenatal 14 days to 21 days postpartum.
Control group is fed to basal diet; The L group is fed and (is contained glycerine 30g approximately, Se 2.6mg and probio 2.07 * 10 previous day to basal diet+product 276.4g/
9Cfu)-the 276.4g corn flour; The M group is fed and (is contained glycerine 60g approximately, Se5.1mg and probio 4.14 * 10 previous day to basal diet+product 552.7g/
9Cfu)-the 552.7g corn flour); The H group is fed and (is contained glycerine 90g approximately, Se7.6mg and 6.21 * 10 previous day to basal diet+product 829.1g/
9Cfu)-the 829.1g corn flour.When feeding ox every afternoon, product is admixed mixed concentrate, throw something and feed separately.
2. test method
Plasma glucose (Glucose) concentration adopts the method for Sigma glucose oxidase enzyme reagent kit to measure; Free fatty (Non-esterified fatty acids, NEFA) concentration adopts Nanjing to build up the method mensuration of the free fatty kit of bio-engineering research institute; Beta-hydroxybutyric acid (β-hydroxybutyrate, BHBA) concentration adopts the method for Sigma kit 310-A to measure; Urine sample employing Bayer CLIN ITEK 500 type Urine Analyzers and supporting test paper, semiquantitative method are measured urine ketone (Urine ketones) concentration.
All milk cows are gathered blood sample respectively at test the 0th, 21 (7d in postpartum), 28 (14d in postpartum), 35 (21d in postpartum) d.All milk cows are gathered urine sample respectively at postpartum the 7th, 14,21d.
3. result of the test
The various dose of feeding perinatal period as shown in Table 1 is rich in the compound probiotic product of glycerine and organic selenium, and product treatment group plasma glucose concentration is higher than control group, and blood plasma free fatty acid and beta-hydroxybutyric acid concentration and urine ketone concentration are lower than control group.Along with the increase of daily ration interpolation product, M group plasma glucose concentration and H group are significantly higher than control group (P＜0.05); M group and H group blood plasma free fatty acid concentration significantly are lower than control group (P＜0.05), and H group blood plasma beta-hydroxybutyric acid concentration significantly is lower than control group (P＜0.05); M and H group urine ketone concentration significantly are lower than control group (P＜0.05).
Test shows, perinatal period the compound probiotic product that is rich in glycerine and organic selenium of feeding can regulate and control the cow in perinatal period energy metabolism impairment effectively.The advantage of this product is: raw material sources are abundant, and equipment requires simple, than the suitability for industrialized production safety non-pollution, need not extract glycerine after the fermentation, greatly reduces production cost.This product has glycerine, organic selenium and probio triple function, can improve the endogenous cycle border of body to ruminant, regulate energetic supersession, regulate gut flora, the antagonism pathogenic microorganism, improve a poor appetite, raising immunity of organisms and premunition are a kind of safety, noresidue, the high novel fodder additive of tiring.
Table 1 is rich in the influence of the compound probiotic product of glycerine and organic selenium to perinatal period milk cow plasma metabolite and urine ketone content
The different lowercase alphabet differentials of colleague's shoulder motes different significantly (P＜0.05), the different capitalizations of same column shoulder motes are represented significant difference (P＜0.05).
Claims (1)
1, a kind of additive for microbe feedstuff that is used for adjusting energy metabolism impairment of flocks and herds during perinatal period, its production method may further comprise the steps:
1) bacterial classification candida krusei (Candida krusei), candida utili (Candida utilis), streptococcus thermophilus (Streptococcus Thermophilus);
2) microbial inoculum is produced culture medium quality volume ratio
15%～25% glucose, 0.1%～1% yeast extract, 0.1%～0.3% urea, 0.1%～0.3% Dried Corn Steep Liquor Powder, 0.3%～0.4% potassium dihydrogen phosphate, 4%～6% sodium chloride, all the other are water;
3) slant activation culture medium
Candida krusei activation CM0357 culture medium: 40g glucose, 2g urea, 4g yeast extract, 20g agar, 1000ml water, pH4.5,115 ℃ of high pressure steam sterilization 15min, standby;
Candida utili, streptococcus thermophilus activation YPD culture medium: 20g glucose, 5g yeast extract, 10g peptone, 20g agar, 1000ml water, initial 6.7,115 ℃ of high pressure steam sterilization 15min of pH, standby;
4) do not add agar in the liquid activation medium slant activation culture medium;
5) sodium selenite aqueous solution of inorganic selenium source preparation selenium content 0.2～2mg/ml, 100～125 ℃ of high pressure steam sterilization 20～30min, standby;
6) adsorbent corn flour;
7) acquisition of the activation of bacterial classification and seed liquor
The candida krusei that is kept at 4 ℃ is inoculated on the CM0357 solid medium, cultivates 30～40h in 20～30 ℃; Switching is 3～5 times so continuously, be inoculated in the CM0357 fluid nutrient medium, and in 20～30 ℃, 150r/min, shaking table is cultivated 24～36h, gets the candida krusei seed liquor, and its cfu reaches 10
9More than/the ml; The candida utili that is kept at 4 ℃ is inoculated on the YPD solid medium, cultivate 30～40h in 20～30 ℃, switching is 3～5 times so continuously, be inoculated in the YPD fluid nutrient medium, in 20～30 ℃, 150r/min, shaking table is cultivated 24～36h, get the candida utili seed liquor, its cfu reaches 10
9More than/the ml; The streptococcus thermophilus that is kept at 4 ℃ is inoculated on the YPD solid medium, cultivates 36～48h in 36～45 ℃; Switching is 3～5 times so continuously, is inoculated in the YPD fluid nutrient medium, in 36～45 ℃, cultivates 24～36h, gets the streptococcus thermophilus seed liquor, and its cfu reaches 10
10More than/the ml;
8) produce
Get and produce culture medium batching glucose 15～25kg, yeast extract 0.1～1.0g, urea 100～300g, Dried Corn Steep Liquor Powder 100～300g, potassium dihydrogen phosphate 300～400g and sodium chloride 4～6kg, it is dissolved in the water, make cumulative volume reach 100L, join then in the 150L fermentation tank, 115～120 ℃ of high pressure steam sterilization 15min, be cooled to 40～50 ℃, inorganic selenium source joined microbial inoculum produce in the culture medium, make that selenium concentration is 2～10mg/kg in the culture medium; Candida krusei, candida utili and streptococcus thermophilus seed liquor were inoculated in microbial inoculum in 13: 1: 2 by volume again and produce in the culture medium, total inoculum concentration volume ratio is 6%～11%, and cultivation temperature is 20～40 ℃, and throughput is 0.05m
3/ h, shaking speed 70r/min, tank pressure are 0.04Mpa, incubation time is 96～132h;
9) product obtains
After cultivating end, nutrient solution mixes with the corn flour of 50kg, and 35～40 ℃ of oven dry promptly obtain a kind of compound probiotic product that is rich in glycerine and organic selenium.
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| CN101869183A (en) * | 2010-05-28 | 2010-10-27 | 广州立达尔生物科技有限公司 | Selenium-enriched yeast hydrolysate and production method thereof |
| CN102174421A (en) * | 2011-03-10 | 2011-09-07 | 南京农业大学 | Glycerol-producing saccharomyces cerevisiae NAU-ZH-GY1 and application thereof |
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| CN1283171C (en) * | 2005-07-12 | 2006-11-08 | 南京农业大学 | Selenium-rich probiotics foder additive production method and its product |
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| CN101869183A (en) * | 2010-05-28 | 2010-10-27 | 广州立达尔生物科技有限公司 | Selenium-enriched yeast hydrolysate and production method thereof |
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| CN102174421A (en) * | 2011-03-10 | 2011-09-07 | 南京农业大学 | Glycerol-producing saccharomyces cerevisiae NAU-ZH-GY1 and application thereof |
| CN102174421B (en) * | 2011-03-10 | 2012-10-17 | 南京农业大学 | Glycerol-producing saccharomyces cerevisiae NAU-ZH-GY1 and application thereof |
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| CN104489305A (en) * | 2014-11-29 | 2015-04-08 | 张旭 | Selenium-rich Chinese herbal medicine type active additive for feed of sheep |
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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 |
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