CN103947904A

CN103947904A - Preparation method of glucose precursor for improving dairy cow rumen flora

Info

Publication number: CN103947904A
Application number: CN201410204205.1A
Authority: CN
Inventors: 邵素英
Original assignee: 邵素英
Current assignee: Beijing Oriental Energy Biotechnology Co., Ltd.
Priority date: 2014-05-15
Filing date: 2014-05-15
Publication date: 2014-07-30
Anticipated expiration: 2034-05-15
Also published as: CN103947904B

Abstract

The invention discloses a preparation method of a glucose precursor for improving dairy cow rumen flora, belonging to the technical field of dairy cow glucose precursors. A microbial preparation and a glucose precursor are taken as main raw materials to be scientifically compounded with Chinese herbal medicine extracts, compound enzymes and other nutrient substances, so that microbial flora needed by the rumen of a dairy cow and endogenous glucose needed for growth and physiology of the dairy cow are effectively replenished and maintained. Experiments prove that the daily milk yield of each dairy cow is increased by 16.81%, the first estrus time of the postpartum dairy cows is shortened by about 10 days, the conception rate of the postpartum dairy cows in the first estrus time is increased by 16.2%, the oestrus rate of the postpartum dairy cow groups is increased by 16%, and the occurrence rate of various diseases of the postpartum dairy cows is averagely reduced by 13.0%-30.5%. According to the preparation method of the glucose precursor disclosed by the invention, high-viscosity glycerol and propylene glycol are uniformly mixed with other materials, so the process is simple, the operation is convenient, the production efficiency is high, the material mixing effect is good and large-scale production can be realized.

Description

A kind of preparation method who improves cow rumen flora glucose precursor

Technical field

The present invention relates to milk cow glucose precursor, particularly a kind of preparation method who improves cow rumen flora glucose precursor.

Background technology

Generally, in ruminant, most of sugar forms VFA (volatile fatty acid) in lumen fermentation process, only has a small amount of glucose to be absorbed in alimentary canal.The ruminant blood sugar of growing up is lower, but and does not mean that the meaning that can ignore glucose metabolism.Glucose, as important trophism monose, is indispensable nutriment in all animal bodies, is undertaking important effect in zooblast metabolism.Glucose supplies is not enough, and ketoacidosis easily occurs ox, and toxaemia easily occurs pregnant sheep, has a strong impact on growth and the health of animal.Therefore, people have strengthened the research to glucose Nutrition and Metabolism.Along with the extensive use of isotope technology, people have had further understanding to the metabolic rule of glucose under status in different nutritional status and physiological situation.At present, international research work mainly concentrates on discussion and the regulation and control of hormone to carbohydrate metabolism to glucose Nutrition and Metabolism rule in each tissue of ruminant, and the research of the domestic Nutrition and Metabolism to ruminant glucose is still in the starting stage.

Ruminant institute energy requirement can not be provided by aliphatic acid separately, in body, some tissue and organ also need glucose especially, as: central nervous system, adipose tissue, embryo and mammary gland, musculature etc., a. embryo growth and newborn generation need a large amount of glucose, sheep latter half of gestation, embryo's glucose metabolism can account for the 40%-70% of ewe glucose total amount; Sheep Ruzhong lactose content is equivalent to 90 times in blood.According to the study, every day, high yield cow synthesized the required glucose of lactose up to 2000g; The sheep galactopoiesis of bosom twin lamb(s) needs 200g glucose every day; The glucose that the synthetic lactose of ruminant that milk yield is high consumes accounts for the 60%-85% of whole body glucose total amount.B. sheep nervous system needs glucose 17g every day, account for the 15%-20% of animal whole body glucose total amount, and human brain will account for 70%-80%, and other ruminant is between the mankind and sheep.C. known often synthetic 1g body fat acid needs 0.65g glucose.Thus, glucose is extremely important to ruminant institute energy requirement.

In ruminant body, the source of glucose is divided into Exogenous Glucose and Inner source glucose, 1. Exogenous Glucose: the solubility sugar and starch in feed through cud almost major part be degraded, directly in alimentary canal, absorbed amount of glucose is very micro-, some starch of cattle and sheep of a large amount of feed fine fodders can be avoided lumen fermentation, enter small intestine and be utilized, the palliating degradation degree of feed starch is relevant with feed kind and processing method.Under grazing condition, the glucose directly absorbing in sheep alimentary canal is almost negligible.It is estimated that, feed roughage is that main sheep only has the glucose of 5-10g feed resource to be absorbed every day; A large amount of glucose of body needs every day will lean on gluconeogenesis synthetic in vivo.2. Inner source glucose: one in maintaining the sheep of state, if can only obtain 5-10g glucose from feed every day, the quantity of the Inner source glucose being synthesized by body gluconeogenesis so can reach 80-120g.Visible, ruminant is not not need or need less glucose, but the source of the glucose obtaining is different from nonruminant.Inner source glucose is the main source of required glucose in ruminant body, and this is one of main feature of ruminant glucose metabolism and utilization.

The same with other ruminant, it is that liver gluconeogenesis increases and surrounding tissue sugar oxidation weakens that milk cow glucose metabolism changes main manifestations in the metabolism of lactation initial stage, to guarantee that glucose can directly enter breast tissue for the synthesis of lactose.Reynokls et al (2003) etc. study discovery, and the net flux of the early stage milk cow internal organs of perinatal period and lactation portal system glucose is zero, even occurs slight negative.First 9 days of the expected date of childbirth to 21 days postpartum, the general output of internal organs glucose raise 267%, and this almost all derives from the gluconeogenesis of liver.The gluconeogenetic main substrate of ruminant liver is that the propionate of lumen fermentation generation is, the glycerine (Seal and Reynokls1993) that the amino acid that lactate, protein metabolism or the internal organ portal system of tricarboxylic acid cycle generation absorb and adipose tissue degraded discharge.From the glucose of propionate, lactate and glycerine heteroplasia, account for 50%-60%, 15%-20% and the 2%-4% (Reynokls et al2003) of the clean burst size of liver glucose perinatal period.Glucose by amino acid heteroplasia at least can reach the 20%-30% of glucose total amount, and wherein the glucose by alanine heteroplasia was 2.3% at antenatal 9 days, and to postpartum, 11d rises to 5.5%.With the above results and consistent, Overton etc. (1998) have reported that the liver of 1d in postpartum is 2 times of antenatal 21d to doctor's ability of alanine.Although quantitatively see that the relative milk yield of amino acid discharging from amino acid pool is unimportant, the above results shows that amino acid can be used as a kind of adaptability substrate synthesizing for commitment glucose in postpartum.

As gluconeogenic precursor, have manyly, the most significant precursor is propionic acid, glycerine, amino acid and lactic acid; Other also has valeric acid, nucleic acid, citric acid and other various organic acids, and because several organic acids such as valeric acid content in blood is very low, therefore, raw sugar effect is limited.

Facts have proved: all high yield cows are always subject to the threat of ketoacidosis in early days to some extent in lactation.Particularly subclinical ketosis, owing to not producing typical clinical symptoms, is difficult to identify, and just shows the decline of cow feeding amount, output of milk reduction, spiritual depressed.But these phenomenons be conventionally interpreted as " newly produce that ox feed intake is low, calving stress, the composite factor such as mismanagement causes ".And the great demand that real reason is supply due to glucose in Contents in Cows when can not meet physiological status and changing fast.In brief, milk cow lacks sugar.

Although do not show what symptom on the milk cow surface of generation subclinical ketosis, the potentiality of giving milk of milk cow are greatly affected conventionally.Recently, U.S.'s Hoard ' s Dairyman magazine finds by the range survey of the U.S., the incidence of disease of various new product ox common diseases from high to low successively: ketoacidosis (clinical type and Subclinical), retained afterbirth, puerperal fever, abomasum displacement and endometritis.The investigation that Canada Onatrio economizes also finds, newly produces ox and goes wrong maximum to be ketoacidosis (clinical type and Subclinical), to be secondly mammitis (clinical type and Subclinical), be then hoof disease, retained afterbirth, milk fever and abomasum displacement.As can be seen here, ketoacidosis (clinical type and Subclinical) is newly to produce the highest disease of the ox incidence of disease in milk cattle cultivating developed country.In China, because relevant enquiry based work is not yet universal, but undeniable, ketoacidosis is one of maximum output of milk " killer " being hidden in inside, cattle farm.

Science, rationally, effectively for milk cow provides metabolizable glucose, can effectively improve the output of milk, reduce the incidence of disease of ketoacidosis and displaced abomasums, effectively improve the early stage dry matter intake of Lactation of Dairy Cow and shorten the calving interval and rutting time first.

Solving milk cow is exactly in feed, to add raw sugared body, i.e. glucose precursor at the effective ways of the early stage glucose supplies deficiency of lactation.

Cow rumen dysbacteriosis is due to irrational raising dairy cattle way to manage, the unreasonable use of antibiotic medicine, cause milk cow alimentary canal, particularly in cud, beneficial microbe reduces or disappears, microbial ecological balance is destroyed and cause that cow rumen digests and assimilates hypodynamia, and autonomic nerve disorder, further causes the gradual nutrient consumption sexual exhaustion of milk cow, finally cause milk cow to eliminate even dead, on veterinary clinic, claim milk cow intractable apositia.The same with other ruminant, one of topmost feature of milk cow can extensively be utilized high-fiber feeding stuff exactly, this feature has benefited from rumen microorganism, and the cellulose in milk cow forage mainly relies on bacterium, infusorian and the fungi in cud to decompose, in cellulosic decomposition digestion, the effect of bacterium and fungi performance 80%, infusorian plays balance control action, and under rumen microorganism synergy, cellulose is decomposed piecemeal, the final volatile fatty acid that produces, is absorbed by milk cow.

Part rumen microorganism is pernicious, and as methane backeria, its Main Function is exactly synthesizing methane; Methane is considered to be only second to the important gas that causes global warming of carbon dioxide, and in Contents in Cows, the generation of methane is also the main cause of fermentation energy loss in cud, and the fodder energy of about 6-15% is depleted with the form of methane.

Therefore, the required beneficial microbe of reasonable supplement cow rumen can reduce the consumption of energy matter, promotes absorption and the metabolism of endogenous glucose.

Chinese patent CN102247570B discloses a kind of Chinese medicine composition for the treatment of ketosis of dairy cows, use this traditional Chinese medicine composition for treating ketosis of dairy cows 58 examples, cure rate reaches 86%, but fundamentally do not solve the problem of the early stage glucose supplies deficiency of lactation, cure the symptoms, not the disease, not good to primary ketoacidosis result for the treatment of.Chinese patent CN101822317B discloses a kind of compound feed additive and preparation method who prevents and treats ketosis of dairy cows.It is by Chinese traditional medicine angelica, the radix paeoniae rubrathe, Ligusticum wallichii, cultivated land, motherwort, costus root, Radix Codonopsis, the Radix Astragali, hawthorn, the extract that dried orange peel is made and sodium propionate, cobaltous sulfate, copper sulphate, manganese sulfate, nicotinic acid, neo dohyfral D3, calcium dihydrogen phosphate, the common compound preparation forming of citric acid, can air making-up and spleen enlivening, blood circulation promoting and enriching, increase glycogen and alleviate acid poisoning, supplement the trace element lacking, mend the sugar phosphorus of replenishing the calcium, reach ketoacidosis is prevented fast and effectively and treated, wherein Chinese medical extract adopts water-boiling method to extract, effective component extraction rate is low, cost is high, glucose precursor is composite incomplete simultaneously, resultant effect is not good, can not finely maintain the microorganism species of cow rumen.Chinese patent CN101305774B discloses a kind of cow compound nutritional supplement at perinatal period, the pre-mixing agent raw material of following composition by weight, consists of: rumen bypass vitamin E:0.5-1 part, cross cud beta carotene: 0.2-0.3 part, cross cud nicotinic acid 5-6 part, cross cud biotin: 0.015-0.02 part, cross cud choline: 3-4 part, lysine-zn 0.35-0.45 part, glucose oxidase preparation: 50-60 part, brown sugar add 100 parts in right amount to.There is the milk cow of raising immunity of organisms, protection hepatic and renal function, can prevent postpartum disease of cow; There is the reproductive performance of improvement, shorten the function in calving interval; There is promotion metabolism, increase the functions such as milk yield, do not solve equally the under-supply problem of the early stage milk cow endogenous glucose of lactation, not ideal to the production performance texts of the preventing of the diseases such as ketoacidosis, treatment and milk cow.Chinese patent CN102499331B cow rumen regulation and control agent and preparation method thereof.Described rumen regulation and control agent is for to cultivate saccharomyces cerevisiae, candida tropicalis and aspergillus oryzae, then the product obtaining after mixed culture fermentation, wherein, the volume ratio of saccharomyces cerevisiae, candida tropicalis and aspergillus oryzae is 0.9～1.1: 1.8～2.2 during fermentation: 0.9～1.1.Advantage is promotion and safeguards microbiota balance in cud, stablize ruminal pH value, alleviate the milk cow acid poison that high fine fodder causes, reduce the concentration of ammonia in cud, promote its utilization to nitrogen, improve its micro-ecological environment, but be mainly by stimulating the breeding of cellulose-decomposing bacteria and lactic acid bacteria in cud to achieve the above object, supplementing targetedly the required beneficial bacterium of cud.

In sum, prepare the glucose precursor that a kind ofly effectively improves cow rumen microorganism species, significantly improves the required endogenous glucose of milk cow necessary.

Summary of the invention

Technical problem solved by the invention is the deficiency that overcomes existing dairy cow nutrition material, for the feed feature of milk cow and digestion, absorption and metabolism characteristic, provide a kind of and improve cow rumen digestion flora, effectively supplement the required endogenous glucose of milk cow growth and physiology to prevent and to treat milk cow intractable apositia and the preparation method of the glucose precursor of glucose and the various diseases that causes for want of.

In order to achieve the above object, the present invention is by the following technical solutions:

A preparation method who improves cow rumen flora glucose precursor, comprises the steps:

Described glucose precursor is prepared by the raw material of following parts by weight:

Microorganism formulation 45-55 part, propane diols 22-28 part, glycerine 22-28 part, calcium propionate 22-28 part, alumino-silicate 22-28 part, Chinese herbal medicine extract 18-22 part, niacinamide 12-18 part, nicotinic acid 12-18 part, sodium lactate 10-15 part, sodium chloride 10-15 part, complex enzyme 8-12 part, citric acid 5-10 part, rumen glucose 5-10 part, rumen-bypass amino acid 3-8 part, crosses cud choline 5-8 part, B B-complex 3-5 part, composite mineral matter 2-4 part.

Described microorganism formulation is occupy cud clostridium (Clostridium ruminocolum) and aspergillus niger (Aspergillus niger) through mixed culture fermentation, low-temperature negative-pressure Vacuum Concentration, fluidized bed drying, low-temperature grinding and make;

Described cud clostridium (Clostridium ruminocolum) bacterial strain that occupies is that the disclosed strain of Chinese patent CN101148651B occupies cud clostridium (Clostridium ruminocolum) H1, and this clostridium has the activity of degraded cellulose and zytase, wood sugar and Ester; Chinese microorganism strain administration committee common micro-organisms center, preservation address on August 6th, 2007, have been preserved in: No. 3, great Tun road, Chaoyang District, BeiJing, China city first, deposit number: CGMCC NO.2125;

Described aspergillus niger (Aspergillus niger) bacterial strain is specially bacterial strain aspergillus niger (Aspergillus niger) Li-2013-03 that produces high activity cellulase.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on July 15th, 2013 and (is called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC NO.7927.

The preparation method of described microorganism formulation comprises the steps:

(1) aspergillus niger CGMCC NO.7927, occupy cud clostridium CGMCC NO.2125 actication of culture

The slant strains of intact aspergillus niger CGMCC NO.7927 is inoculated in to slant medium, cultivates 36-48h for 28-33 ℃ and carry out actication of culture, so activate 2-4 time;

The intact slant strains that occupies cud clostridium CGMCC NO.2125 is inoculated in to slant medium, cultivates 24-36h for 35-41 ℃ and carry out actication of culture, so activate 2-3 time;

Described aspergillus niger CGMCC NO.7927 slant medium consists of: potato (liquor) 200g, glucose 20g, agar 20g, Chinese herbal medicine extract 5-20g, distilled water l000mL, 5.8,121 ℃ of sterilizing 20min of pH value;

The described cud clostridium CGMCC NO.2125 slant medium that occupies consists of: beef extract 3g, sodium chloride 5g, peptone 10g, agar 15g, Chinese herbal medicine extract 5-10g, distilled water l000mL, 6.8,121 ℃ of sterilizing 20min of pH value;

(2) aspergillus niger CGMCC NO.7927 liquid seeds expands cultivation

1. first order seed is cultivated: the aspergillus niger CGMCC NO.7927 slant strains after step (1) is activated is with spore under aseptic washing, access in 500 ml shake flasks, 100 milliliters of liquid seed culture medium loading amounts, 28-33 ℃, 80-120rpm shaking table cultivation 36-48h;

2. secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum concentration access of 10%, condition of culture is identical with first order seed;

3. three grades of seed culture: by secondary seed with in 5000 milliliters of three grades of seed shaking flasks of 8% inoculum concentration access, 1000 milliliters of fluid nutrient medium loading amounts, 28-33 ℃, 80-120rpm shaking table are cultivated 36-48h;

4. first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 8% inoculum concentration access total measurement (volume) is 150L, fermentation medium loading amount 100L, controlling pH value is 5-6, cultivation temperature 28-30 ℃, mixing speed 200-400rpm, ventilation (V/V) 1:0.8-1.2, incubation time 36-48h, dissolved oxygen 10-20%;

Described aspergillus niger CGMCC NO.7927 one-level, secondary, three grades of seed culture mediums consist of: potato (liquor) 200g, glucose 20g, Chinese herbal medicine extract 15-20g, trehalose 10-30g, distilled water l000mL, 5.5,121 ℃ of sterilizing 20min of pH value;

Described aspergillus niger CGMCC NO.7927 first class seed pot culture medium consists of: corn flour 50-60g, bean powder 15-25g, wheat bran 10-15g, Chinese herbal medicine extract 15-20g, trehalose 10-30g, ammonium sulfate 1-3g, dipotassium hydrogen phosphate 1-2g, potassium dihydrogen phosphate 1-2g, pure water l000mL, pH value 5-7,121 ℃ of sterilizing 20min;

Described aspergillus niger CGMCC NO.7927 first class seed pot zymotic fluid cell concentration is 7.0x10 ⁹-8.0x10 ⁹individual/ml;

(3) occupy cud clostridium CGMCC NO.2125 liquid seeds and expand cultivation

1. first order seed is cultivated: the cud clostridium CGMCC NO.2125 slant strains that occupies after step (1) activation is accessed in 500 ml shake flasks to 100 milliliters of culture medium loading amounts, 180 revs/min of rotary shaking tables, cultivation temperature 35-41 ℃, incubation time 10-15h;

3. three grades of seed culture: by secondary seed with in 5000 milliliters of three grades of seed shaking flasks of 10% inoculum concentration access, 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, cultivation temperature 35-41 ℃, incubation time 10-15h;

4. first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 10% inoculum concentration access total measurement (volume) is 150L, fermentation medium loading amount 100L, cultivation temperature 37-39 ℃, mixing speed 200-400rpm, ventilation (V/V) 1:1-2, tank pressure 0.05Mpa, incubation time 10-20h;

The described cud clostridium CGMCC NO.2125 one-level, secondary, three grades of seed culture medium weight of occupying consists of:

Dusty yeast 0.3-0.5%, glucose 1-1.5%, peptone 0.3-0.5%, beef extract 0.5-0.8%, dipotassium hydrogen phosphate 0.8-1.5%, Chinese herbal medicine extract 1.5-2%, trehalose 1-3%, insufficient section pure water is supplied, pH value 6.8,121-123 ℃ of sterilizing 30-40min.

The described cud clostridium CGMCC NO.2125 seed tank culture basic weight amount that occupies consists of:

Maltodextrin 5-15%, dusty yeast 0.4-0.8%, Chinese herbal medicine extract 1.5-2%, trehalose 1-3%, peptone 0.1-0.5%, corn steep liquor 0.1-0.5%, dipotassium hydrogen phosphate 0.8-1.5%, magnesium sulfate 0.05-0.1%, insufficient section pure water is supplied, pH value 6.8,121-123 ℃ of sterilizing 30-40min.

It is described that to occupy cud clostridium CGMCC NO.2125 first class seed pot zymotic fluid cell concentration be 7.0x10 ⁹-8.0x10 ⁹individual/ml;

(4) aspergillus niger CGMCC NO.7927, occupy cud clostridium CGMCC NO.2125 first class seed pot liquid seeds and mix

The cud clostridium CGMCC NO.2125 seeding tank liquid seeds that occupies that the aspergillus niger CGMCC NO.7927 seeding tank liquid seeds that step (2) is obtained obtains with step (3) evenly mixes by volume;

Described volume ratio 1:0.5-1;

Described mixing material seed cell concentration is: 7.0x10 ⁹-8.0x10 ⁹individual/ml;

(5) secondary seed tank alternate

Mixed uniformly first class seed pot liquid seeds in step (4) be take to the secondary seed tank that 10% inoculum concentration access total measurement (volume) is 300L, alternate culture medium loading amount 240L, secondary seed tank fermentation tank, cultivation temperature 33-39 ℃, mixing speed 200-700r/m, ventilation (V/V) 1:1-3, incubation time 15-24h;

Described alternate culture medium consists of: maltodextrin 50-150g, corn flour 50-60g, bean powder 15-25g, wheat bran 10-15g, Chinese herbal medicine extract 15-20g, trehalose 10-30g, dusty yeast 4-8g, corn steep liquor 1-5g, ammonium sulfate 1-3g, dipotassium hydrogen phosphate 1-2g, potassium dihydrogen phosphate 1-2g, pure water l000mL, pH value 5-7,121 ℃ of sterilizing 20min;

Described secondary seed tank zymotic fluid cell concentration is 7.0x10 ⁹-8.0x10 ⁹individual/ml;

(6) bacterial classification mixed culture fermentation

Secondary seed tank alternate liquid seeds in step (5) is accessed to fermentation tank, cultivation temperature 28-30 ℃, mixing speed 200-700r/m, ventilation (V/V) 1:1-3, incubation time 15-20h with 6% inoculum concentration; Then with 1-2 ℃/h rate of temperature fall slow cooling to 10-15 ℃, constant temperature culture 10-12h; Continuation to 2-5 ℃, now, is appended access fermentation tank, constant temperature culture 6-8h by secondary seed tank alternate liquid seeds in step (5) with 4% inoculum concentration with 1-2 ℃/h rate of temperature fall slow cooling; Finally with 1-2 ℃/h heating rate, be slowly warming up to 10-15 ℃, constant temperature culture 10-12h; Continuation is slowly warming up to 37-39 ℃, constant temperature culture 15-20h with 1-2 ℃/h heating rate;

Described fermentation medium consists of: maltodextrin 50-150g, corn flour 50-60g, bean powder 15-25g, wheat bran 10-15g, Chinese herbal medicine extract 30-50g, trehalose 30-40g, dusty yeast 4-8g, corn steep liquor 1-5g, ammonium sulfate 1-3g, dipotassium hydrogen phosphate 1-2g, potassium dihydrogen phosphate 1-2g, pure water l000mL, pH value 5-7,121 ℃ of sterilizing 20min;

Described ferment tank liquid cell concentration is 7.0x10 ⁹-8.0x10 ⁹individual/ml;

(7) zymotic fluid is concentrated in vacuo to 45% of original volume through low-temperature negative-pressure, obtains bacterium concentrate, and concentrate evenly mixes for 0.5:1 in mass ratio with carrier, in 45 ℃ of fluidized bed dryings, and 40 ℃ of low-temperature grinding, grinding particle size 0.5-1.5mm, obtains microorganism formulation;

Described vehicle weight umber consists of: CaCO ₃20-23 part, dextrin 10-12 part.

Described microorganism formulation viable count content is 1.5x10 ¹⁰-3x10 ¹⁰individual/g.

Described glycerine is food stage glycerine, purity 99.5%;

Described alumino-silicate is for having selective strong adsorbing alumino-silicate or alumino-silicate derivatives, composite with Chinese herbal medicine extract science, the multiple toxin such as energy adsorption of aflatoxin, vomitoxin, zearalenone, T-2 toxin, also tool strengthens immunity, protects the effects such as the strong kidney of liver, and the nutrition such as amino acid, vitamin, trace element in can adsorption feed;

Described Chinese herbal medicine extract is prepared by the raw material of following parts by weight: Radix Angelicae Sinensis 55-65 part, Ligusticum wallichii 55-65 part, fructus amomi 50-60 part, radix paeoniae rubrathe 50-60 part, cultivated land 50-60 part, Divine Comedy 45-55 part, Fructus Hordei Germinatus 45-55 part, motherwort 45-55 part, costus root 40-45 part, Radix Codonopsis 40-45 part, rhizoma atractylodis 40-45 part, Radix Astragali 40-45 part, honeysuckle 40-45 part, cordate houttuynia 35-45 part, Poria cocos 35-45 part, jujube kernel 35-45 part, fushen 35-45 part, polygala root 30-40 part, fry fennel 30-40 part, bighead atractylodes rhizome 30-40 part, bark of official magnolia 30-40 part, Fructus Aurantii 25-35 part, Radix Glycyrrhizae 25-35 part, hawthorn 25-35 part, dried orange peel 25-35 part.

The preparation method of described Chinese herbal medicine extract is: respectively said herbal medicine being crushed to particle diameter is below 2 millimeters, then in container, evenly mix and add the water of 3-6 times of weight, control temperature 70 C-90 ℃ and keep 2-4h, then be cooled to 45-60 ℃, add the mixing enzyme preparation of mixed material gross weight 5-10% to carry out enzymolysis, with newborn acid for adjusting pH value, be 5.5-6.8, enzymolysis 2-4h, finally add the mixture of mixed material 0.5-3 times of weight ethanol and propyl alcohol, control temperature to 60 ℃-78 ℃ of maintenance 3-4h, filter; It is more than 20% that filtrate decompression is concentrated into solid content, and then freeze drying, 40 ℃ of low-temperature grinding obtain Chinese herbal medicine extract, grinding particle size 0.5-2mm;

The parts by weight of described mixed enzyme consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, beta-glucosidase 10-15 part, zytase 15-20 part, pentosanase 15-20 part, Pullulanase 20-30 part, beta amylase 10-15 part, neutral proteinase 10-15 part, acid protease 10-15 part, superoxide dismutase 5-10 part, glucose oxidase 5-10 part, acid phosphatase 5-10 part;

The mass ratio that described ethanol and propyl alcohol mix is 1:1-2, described concentration of alcohol >=75%;

Described sodium lactate is 60% food stage sodium lactate;

Described complex enzyme is evenly mixed by the solid enzyme preparation of following parts by weight: neutral proteinase 22-28 part, acid protease 22-28 part, alkalescent xylanase 22-28 part, acidic xylanase 22-28 part, cellulase 22-28 part, 1,4 beta-glucanase 15-20 part, pectase 15-20 part, amylase 15-20 part, seminase 10-15 part, phytase 10-15 part.

Described citric acid is solid citric acid;

Described per kilogram B B-complex comprises dehydroretinol 800mg, orotic acid 000mg, Catergen 000mg, riboflavin 1600mg, pantothenic acid 1900mg, pyridoxamine 1800mg, cobalamin 1200mg, inositol 5000mg, vitamin E 600mg, neo dohyfral D3 5500mg, prokayvit 4000mg; Its surplus is carrier, and described carrier is rice bran.

Described per kilogram composite mineral matter comprises calcium 6000mg, phosphorus 4200mg, sodium 5100mg, copper 800mg, iron 1800mg, potassium 3800mg, manganese 1400mg, cobalt 1600mg, iodine 5000mg, magnesium 1450mg; Its surplus is carrier, and described carrier is rice bran.

Preparation method:

1) according to above-mentioned formula rate, accurately take each raw material, first add respectively its quality 0.5-1 sterilized water doubly to dilute propane diols and glycerine, then dilution is mixed, heating water bath is to 30-50 ℃ simultaneously, limit is uniformly mixed to obtain mixture A, heat preservation for standby use;

2) calcium propionate, alumino-silicate, sodium lactate, sodium chloride, citric acid are evenly mixed to get to mixture B;

3) Chinese herbal medicine extract, rumen glucose, rumen-bypass amino acid, mistake cud choline, composite mineral matter are evenly mixed to get to mixture C;

4) microorganism formulation, niacinamide, nicotinic acid, complex enzyme, B B-complex are evenly mixed to get to mixture D;

5) mixture B is added to LDH-1 type coulter type mixer, start mixer, then add mixture A evenly to mix 10-20min; Then add mixture C, evenly mix 5-10min; Finally add mixture D, evenly mix 3-5min, final mixture is sealed, packs and obtain the glucose precursor that can improve cow rumen flora.

Using method: 1. add in fine fodder, coarse fodder or premix, mix;

2. this product is added to 10-20 times of running water, oral or be sprayed on TRM after evenly mixing;

Use amount: newly produce ox: 100-120g/ head, sky;

Cow in milk: 80-100g/ head, sky;

Late drying period ox: 100-120g/ head, sky;

Beef cattle: 20-25kg/ ton daily ration;

Operational phase:

1. improve production performance: antenatal 20 days to 30 days postpartum;

2. prophylactic treatment ketoacidosis: postpartum prevention and health care: the 1st day to the 7th day postpartum;

Postpartum therapy ketoacidosis: measure blood ketone content postpartum, gavage above if content reaches 1.2mmol/L, gavage continuously 5 days;

The present invention occupies cud clostridium bacterial strain CGMCC NO.2125 can degraded cellulose and xylan, wood sugar and ester class, the complex enzyme system that this bacterium contains degraded cellulose, comprise circumscribed cellulase, endo cellulase, zytase, mannonase esterase, pectase, enzyme work is respectively 4.1 ± 1.19U/mg, 118.7 ± 4.5U/mg, 167.36 ± 22.9U/mg, 45.9 ± 3.52U/mg, 4 ± 0.5U/mg, 35.3 ± 1.77U/mg, the suitableeest cultivation temperature 35-41 ℃ of this bacterium, optimum pH 6.5-7.5.

Aspergillus niger strain CGMCC No.7927 of the present invention has following microbial characteristic: 1, morphological feature: biology morphology comprises conidium, born of the same parents' stalk, top capsule, produces several parts such as born of the same parents' structure.Conidial head is spherical to Radiation, diameter 150-450 μ m, and conidiophore betides matrix.Born of the same parents obstruct stem 1000-3000 (length) * 12-20 (diameter) μ m, yellow or yellowish-brown, and wall is level and smooth; Spherical or the almost spherical of top capsule, diameter 45-75 μ m, surface can be educated comprehensively; Produce born of the same parents' structure double-deck, metulae 10-20 (length) * 4.5-7.0 (diameter) μ m, bottle stalk 6-10 (length) * 2.5-3.5 (diameter) μ m, conidium is spherical or subsphaeroidal, diameter 3-4.5 μ m, brown, wall is coarse.2, cultivate and learn feature: bacterial strain is grown rapidly on wort agar culture medium, and 28 ℃ of 4 days spores can be paved with inclined-plane; Quality velvet shape or be slightly with cotton-shaped; Conidium structure is a large amount of, and brown-black, without diffusate; Bacterium colony reverse side is slightly yellow.3, physiological and biochemical property: Aspergillus niger strain CGMCC No.7927 can grow in the carbon sources such as maize straw, straw, wood chip, potato, corn flour, soluble starch, molasses, optimal pH scope 5-6, optimum growth temperature scope 28-33 ℃, the suitableeest product enzyme temperature range 28-30 ℃.

The triage techniques route of Aspergillus niger strain CGMCC No.7927 of the present invention is: experiment (fermenting property mensuration) is measured → expanded to the preparation → mutagenic treatment → plate isolation → primary dcreening operation of starting strain → inclined-plane cultivation → spore suspension → multiple sieve → genetic stability.

Press mutagenesis screening scheme, mutant strain step-sizing is eliminated, finally to strain excellent through fermenting property test screen, obtain a plant height and produce enzyme performance bacterial strain black-koji mould Aspergillus niger Li-2013-03, circumscribed 1,4 beta-glucanase, Endo-β-glucanase, beta-glucosidase and the filter paper enzyme activity of cellulase after 96 hours that ferment reaches respectively 620U/mL, 1289U/mL, 456U/mL and 732U/mL.

Beneficial effect:

1. the glucose precursor that prepared by the present invention is for the feed feature of milk cow and digestion, absorption and metabolism characteristic, take microorganism formulation and glucose precursor is primary raw material, the composite Chinese herbal medicine extract of science, complex enzyme and other milk cow desired nutritional material, not only supplement and maintained the required microorganism species of cow rumen, guarantee the normal function of cud, promoted and improved digestibility and the absorptivity of nutriment; And for milk cow provides the abundantest, the most comprehensive nutriment of production, growth and prevention, treatment disease, particularly effectively supplemented milk cow growth and the required endogenous glucose of physiology, effectively maintain milk cow human body energy balance.

2. the glucose precursor that prepared by the present invention can effectively improve the output of milk, and every cow head daily yielding can improve 16.81%; Effectively shorten nearly 10 days of the time of milk cow postpartum heat, improve the conception rate 16.2% of the first feelings phase of postpartum milk cow, increase the ratio of oestrusing 16% of Postpartum Cows colony, effectively improve the reproductive performance of milk cow; Can significantly reduce the incidence of the diseases such as milk cow intractable in postpartum apositia, ketoacidosis, mammitis, endometritis, cud displacement, puerperal fever and retention of afterbirth, the average 13.0-30.5% that reduces, t checks significant difference (P < 0.05).

3. Chinese herbal medicine extract of the present invention adopts enzymolysis and alcohol precipitation to extract, and has extracted to greatest extent and has planted the active ingredient in herbal medicine, has improved raw material availability, has reduced production cost.And, Chinese herbal medicine extract of the present invention is applied in the preparation method of microorganism formulation, improved the resistance of rumen microorganism to Chinese herbal medicine, for the normal operation of later stage digestive system, establish solid foundation, overcome the defect that because of application Chinese herbal medicine, cud beneficial microbe is suppressed or destroyed in prior art simultaneously; The preparation method of microorganism formulation of the present invention to microorganism have anti-stress and rejuvenation effect, microorganism in microorganism formulation has possessed the resistance to the action of a drug to Chinese herbal medicine effective ingredients, and the Chinese herbal medicine effective ingredients that the later stage feeds can not suppress and affect normal growth and the breeding of adding microorganism.

4. alumino-silicate of the present invention has selective absorption function and powerful suction-operated, only adsorb the mycotoxin producing because of mould in milk cow feed, do not adsorb microorganism and nutriment, improved the security of cow producing milk and the various diseases that effectively prevents mycotoxin to cause.

5. preparation method of the present invention, glycerine and propane diols that viscosity is higher evenly mix with other material, and technique is simple, easy to operate, production efficiency is high, mixing of materials is effective, can accomplish scale production.

The specific embodiment

Below by specific embodiment narration the present invention.Unless stated otherwise, in the present invention, technological means used is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, but not limits the scope of the invention, and the spirit and scope of the invention are only limited by claims.To those skilled in the art, do not deviating under the prerequisite of essence of the present invention and scope various changes that the material component in these embodiments and consumption are carried out or change and also belong to protection scope of the present invention.

The preparation of embodiment 1 microorganism formulation

The slant strains of intact aspergillus niger CGMCC NO.7927 is inoculated in to slant medium, cultivates 24h for 30 ℃ and carry out actication of culture, so activate 3 times;

The intact slant strains that occupies cud clostridium CGMCC NO.2125 is inoculated in to slant medium, cultivates 24-36h for 39 ℃ and carry out actication of culture, so activate 2 times;

Described aspergillus niger CGMCC NO.7927 slant medium consists of: potato (liquor) 200g, glucose 20g, agar 20g, Chinese herbal medicine extract 10g, distilled water l000mL, 5.8,121 ℃ of sterilizing 20min of pH value;

The described cud clostridium CGMCC NO.2125 slant medium that occupies consists of: beef extract 3g, sodium chloride 5g, peptone 10g, agar 15g, Chinese herbal medicine extract 8g, distilled water l000mL, 6.8,121 ℃ of sterilizing 20min of pH value;

(2) aspergillus niger CGMCC NO.7927 liquid seeds expands cultivation

1. first order seed is cultivated: the aspergillus niger CGMCC NO.7927 slant strains after step (1) activation, with spore under aseptic washing, is accessed in 500 ml shake flasks to 100 milliliters of liquid seed culture medium loading amounts, 30 ℃, 100rpm shaking table cultivation 24h;

3. three grades of seed culture: by secondary seed with in 5000 milliliters of three grades of seed shaking flasks of 8% inoculum concentration access, 1000 milliliters of fluid nutrient medium loading amounts, 30 ℃, 100rpm shaking table are cultivated 24h;

4. first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 8% inoculum concentration access total measurement (volume) is 150L, and fermentation medium loading amount 100L, controlling pH value is 5.5,30 ℃ of cultivation temperature, mixing speed 300rpm, ventilation (V/V) 1:1.2, incubation time 42h, dissolved oxygen 15%;

Described aspergillus niger CGMCC NO.7927 one-level, secondary, three grades of seed culture mediums consist of: potato (liquor) 200g, glucose 20g, Chinese herbal medicine extract 15g, trehalose 20g, distilled water l000mL, 5.5,121 ℃ of sterilizing 20min of pH value;

Described aspergillus niger CGMCC NO.7927 first class seed pot culture medium consists of: corn flour 55g, bean powder 20g, wheat bran 12g, Chinese herbal medicine extract 18g, trehalose 20g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, pure water l000mL, 5.5,121 ℃ of sterilizing 20min of pH value;

Described aspergillus niger CGMCC NO.7927 first class seed pot zymotic fluid cell concentration is 8.0x10 ⁹individual/ml;

(3) occupy cud clostridium CGMCC NO.2125 liquid seeds and expand cultivation

1. first order seed is cultivated: the cud clostridium CGMCC NO.2125 slant strains that occupies after step (1) activation is accessed in 500 ml shake flasks to 100 milliliters of culture medium loading amounts, 180 revs/min of rotary shaking tables, 39 ℃ of cultivation temperature, incubation time 12h;

3. three grades of seed culture: by secondary seed with in 5000 milliliters of three grades of seed shaking flasks of 10% inoculum concentration access, 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, 39 ℃ of cultivation temperature, incubation time 12h;

4. first class seed pot is cultivated: three grades of seeds be take to the first class seed pot that 10% inoculum concentration access total measurement (volume) is 150L, fermentation medium loading amount 100L, 39 ℃ of cultivation temperature, mixing speed 300rpm, ventilation (V/V) 1:1, tank pressure 0.05Mpa, incubation time 15h;

Dusty yeast 0.4%, glucose 1.5%, peptone 0.4%, beef extract 0.6%, dipotassium hydrogen phosphate 1.2%, Chinese herbal medicine extract 1.5%, trehalose 2%, insufficient section pure water is supplied, 6.8,121 ℃ of sterilizing 30min of pH value.

Maltodextrin 10%, dusty yeast 0.6%, Chinese herbal medicine extract 1.6%, trehalose 2%, peptone 0.3%, corn steep liquor 0.3%, dipotassium hydrogen phosphate 1.2%, magnesium sulfate 0.08%, insufficient section pure water is supplied, 6.8,121 ℃ of sterilizing 30min of pH value.

It is described that to occupy cud clostridium CGMCC NO.2125 first class seed pot zymotic fluid cell concentration be 8.0x10 ⁹individual/ml;

Described volume ratio 1:1;

Described mixing material seed cell concentration is: 8.0x10 ⁹individual/ml;

(5) secondary seed tank alternate

Mixed uniformly first class seed pot liquid seeds in step (4) be take to the secondary seed tank that 10% inoculum concentration access total measurement (volume) is 300L, alternate culture medium loading amount 240L, secondary seed tank fermentation tank, 36 ℃ of cultivation temperature, mixing speed 500r/m, ventilation (V/V) 1:1, incubation time 20h;

Described alternate culture medium consists of: maltodextrin 100g, corn flour 55g, bean powder 20g, wheat bran 12g, Chinese herbal medicine extract 18g, trehalose 20g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, pure water l000mL, 6,121 ℃ of sterilizing 20min of pH value;

Described secondary seed tank zymotic fluid cell concentration is 8.0x10 ⁹individual/ml;

(6) bacterial classification mixed culture fermentation

Secondary seed tank alternate liquid seeds in step (5) is accessed to fermentation tank, 30 ℃ of cultivation temperature, mixing speed 500r/m, ventilation (V/V) 1:1, incubation time 18h with 6% inoculum concentration; Then with 2 ℃/h rate of temperature fall slow cooling to 12 ℃, constant temperature culture 10h; Continuation, with 2 ℃/h rate of temperature fall slow cooling to 4 ℃, now, is appended access fermentation tank, constant temperature culture 6h by secondary seed tank alternate liquid seeds in step (5) with 4% inoculum concentration; Finally with 2 ℃/h heating rate, be slowly warming up to 12 ℃, constant temperature culture 10h; Continuation is slowly warming up to 38 ℃ with 2 ℃/h heating rate, constant temperature culture 20h;

Described fermentation medium consists of: maltodextrin 100g, corn flour 55g, bean powder 20g, wheat bran 12g, Chinese herbal medicine extract 40g, trehalose 35g, dusty yeast 6g, corn steep liquor 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 2g, potassium dihydrogen phosphate 2g, pure water l000mL, 6,121 ℃ of sterilizing 20min of pH value;

Described ferment tank liquid cell concentration is 8.0x10 ⁹individual/ml;

(7) zymotic fluid is concentrated in vacuo to 45% of original volume through low-temperature negative-pressure, obtains bacterium concentrate, and concentrate evenly mixes for 0.5:1 in mass ratio with carrier, in 45 ℃ of fluidized bed dryings, and 40 ℃ of low-temperature grinding, grinding particle size 1mm, obtains microorganism formulation;

Described vehicle weight umber consists of: CaCO ₃22 parts, 12 parts, dextrin.

The microorganism formulation viable count content of preparing through said method is 3x10 ¹⁰individual/g.

The preparation of embodiment 2 Chinese herbal medicine extracts

Described Chinese herbal medicine extract is prepared by the raw material of following parts by weight: 60 parts of Radix Angelicae Sinensis, 60 parts of Ligusticum wallichiis, 55 parts of fructus amomis, 55 parts of the radix paeoniae rubrathe, 55 parts, cultivated land, 50 parts of Divine Comedy, 50 parts, Fructus Hordei Germinatus, 50 parts of motherworts, 42 parts of costus root, 42 parts of Radix Codonopsis, 42 parts of rhizoma atractylodis, 42 parts of the Radixs Astragali, 42 parts of honeysuckles, 40 parts of cordate houttuynias, 40 parts, Poria cocos, 40 parts of jujube kernels, 40 parts of fushens, 35 parts of polygala roots, fry 35 parts, fennel, 35 parts of the bighead atractylodes rhizomes, 35 parts of the barks of official magnolia, 30 parts of Fructus Aurantiis, 30 parts, Radix Glycyrrhizae, 30 parts of hawthorn, 30 parts of dried orange peels.

The preparation method of described Chinese herbal medicine extract is: respectively said herbal medicine being crushed to particle diameter is below 2 millimeters, then in container, evenly mix and add the water of 5 times of weight, control 80 ℃ of temperature and keep 3h, then be cooled to 50 ℃, add the mixing enzyme preparation of mixed material gross weight 8% to carry out enzymolysis, with newborn acid for adjusting pH value, be 6.2, enzymolysis 3h, finally add the mixture of 2 times of weight ethanol of mixed material (95%) and propyl alcohol, the mass ratio that described ethanol and propyl alcohol mix is 1:1.5, control temperature to 70 ℃ and keep 4h, filter; It is 25% that filtrate decompression is concentrated into solid content, and then freeze drying, 40 ℃ of low-temperature grinding obtain Chinese herbal medicine extract;

The parts by weight of described mixed enzyme consist of: 15 parts of endo-beta-glucanases, 15 parts of outer 1,4 beta-glucanases, 12 parts of beta-glucosidases, 18 parts of zytases, 18 parts of pentosanases, 25 parts of Pullulanases, 12 parts of beta amylases, 12 parts of neutral proteinases, 12 parts of acid proteases, 8 parts of superoxide dismutases, 8 parts of glucose oxidases, 8 parts of acid phosphatases;

The Chinese herbal medicine extract granularity of preparing through above-mentioned preparation method is 1.2mm, and effective component extraction rate reaches more than 95.

Embodiment 3 complex enzymes composite

Described complex enzyme is evenly mixed by the solid enzyme preparation of following parts by weight: 25 parts of neutral proteinases, 25 parts of acid proteases, 25 parts of alkalescent xylanases, 25 parts of acidic xylanases, 25 parts of cellulases, 18 parts of 1,4 beta-glucanases, 18 parts of pectases, 18 parts of amylase, 12 parts of seminases, 12 parts of phytases.

Microorganism formulation, Chinese herbal medicine extract and complex enzyme prepared by the embodiment 1-3 of take is respectively raw material, with commercially available propane diols, glycerine, calcium propionate, alumino-silicate, niacinamide, nicotinic acid, sodium lactate, sodium chloride, citric acid, rumen glucose, rumen-bypass amino acid, cud choline, B B-complex, composite mineral matter carry out composite excessively, preparation can improve the glucose precursor of cow rumen flora, concrete composition of raw materials is in Table 1, and preparation method is shown in embodiment 4-6;

Described glycerine is food stage glycerine, purity 99.5%;

Described alumino-silicate is for having selective strong adsorbing aluminate and the derivative thereof crossed;

Described sodium lactate is 60% food stage sodium lactate;

Described citric acid is solid citric acid;

Table 1 composition of raw materials

Embodiment 4

A preparation method who improves the glucose precursor of cow rumen flora, comprises the steps:

1) according to above-mentioned formula rate, accurately take each raw material, first add respectively the sterilized water of 0.5 times of its quality to dilute propane diols and glycerine, then dilution is mixed, while heating water bath to 30 ℃, limit is uniformly mixed to obtain mixture A, heat preservation for standby use;

5) mixture B is added to LDH-1 type coulter type mixer, start mixer, then add mixture A evenly to mix 10min; Then add mixture C, evenly mix 5min; Finally add mixture D, evenly mix 3min, final mixture is sealed, packs and obtain the glucose precursor that can improve cow rumen flora.

Embodiment 5

1) according to above-mentioned formula rate, accurately take each raw material, first add respectively the sterilized water of 0.8 times of its quality to dilute propane diols and glycerine, then dilution is mixed, while heating water bath to 40 ℃, limit is uniformly mixed to obtain mixture A, heat preservation for standby use;

5) mixture B is added to LDH-1 type coulter type mixer, start mixer, then add mixture A evenly to mix 15min; Then add mixture C, evenly mix 8min; Finally add mixture D, evenly mix 4min, final mixture is sealed, packs and obtain the glucose precursor that can improve cow rumen flora.

Embodiment 6

1) according to above-mentioned formula rate, accurately take each raw material, first add respectively the sterilized water of 1 times of its quality to dilute propane diols and glycerine, then dilution is mixed, while heating water bath to 50 ℃, limit is uniformly mixed to obtain mixture A, heat preservation for standby use;

5) mixture B is added to LDH-1 type coulter type mixer, start mixer, then add mixture A evenly to mix 20min; Then add mixture C, evenly mix 10min; Finally add mixture D, evenly mix 5min, final mixture is sealed, packs and obtain the glucose precursor that can improve cow rumen flora.

The glucose precursor application test of embodiment 7 embodiment 5 preparations

1 materials and methods

1.1, test material

1.1.1, product of the present invention

1.1.2, test milk cow: test is carried out a certain cattle farm in Miyun Region of Beijing, and kind is the above Diseases of Cow of china holstein cows 2 tire, and totally 46, wherein 2 tire is 16,3 12, tires, 4 10, tires, 58, tires.Test ox, by parity, the identical or close pair principle of the output of milk in early stage, is divided into experimental group and control group, 23 every group.A upper parity milk cow annual output of milk experimental group 8454kg, control group 8533kg.

1.2, test daily ration forms: test group and control group adopt same feeding and management method.Daily ration consists of fine fodder (corn 52.7%, wheat bran 8%, dregs of beans 7%, cotton dregs 9%, peanut meal 9%, corn DDGS 8%, premix 7115A6%), and coarse fodder is corn silage, sheep's hay, the beans stalks of rice, wheat, etc., clover etc.

1.3, test method: according to milk cow breeding record, be divided at random test group and two processing of control group entering the expected date of childbirth front 15 longicorns.Control group is fed according to a conventional method, test group is except taking same method feeds with control group, and from antenatal 20 days to 30 days postpartum, every day, every cow head was taken this preparation 100g, every this preparation of 100g is a package metro-measuring, and every natural gift are fed to compound nutritional pre-mixing agent perinatal period for 2 times.All for the fine fodder 4-8kg of giving milk of autogamy in examination Cow-feeding field, adjust corn silage quantitative feeding 20kg/ days, roughage (sheep's hay, the beans stalks of rice, wheat, etc., clover) free choice feeding according to raising stage and the output of milk.

1.4, production, metabolism, reproductive performance record: the output of milk of measuring and record every cow head in during 120 days postpartum; Observe, evaluate milk cow body condition; Detect milk cow blood plasma part metabolite; Detect cow serum immunoglobulin content; Detect reproductive performance; The observed and recorded rutting situation of milk cow of participating in the experiment in during 120 days postpartum, whether gestation is as the criterion with the examination per rectum in latter 60 days of breeding.

1.5, disease surveillance and record: Dairy Cows diseases such as () mammitis, endometritis, ketoacidosis, cud displacement, intractable apositia, puerperal fever and retentions of afterbirth is detected, the same day was monitored retention of afterbirth incidence in childbirth, and definition afterbirth not discharge person in 12-24 hour is retention of afterbirth.Adopt ketone powder method to detect ketosis of dairy cows, adopt CMT-Test reagent to detect mammitis.

1.6, statistical analysis is processed initial data with EXCEL software, then uses SAS[11] method carries out variance analysis.

1.7, test period experimental period, lasts 214 days from end of day in November 10 in-2011 on April 10th, 2011.

2. result of the test

2.1 impacts of glucose precursor on milk cow body condition:

Table 2 milk cow 5 part Body Condition Score standards

Score value	Vertebra portion	Flank	Buttocks both sides	Root of the tail both sides	Hipbone, ischium joint
						1	Very outstanding	Root root is visible	Excessive settlement	Crypts is very dark	Very outstanding
2	Obviously outstanding	Most visible	Obviously sink	Crypts is obvious	Obviously outstanding
						3	Outstanding a little	Minority is visible	Sagging a little	Crypts is slightly aobvious	Outstanding a little
4	Straight	Lose completely	Straight	Crypts is not aobvious	Do not show outstanding
						5	Plentiful	Plentiful	Plentiful	Plentiful	Plentiful

Table 3 milk cow body condition visual examination appraisal result

Above result shows: in Diseases of Cow TMR daily ration, add the trend that 100g/ (d head) glucose precursor is significantly improved to milk cow Body Condition Score, can improve milk cow to the digestion of nutriment, absorption and metabolism, be conducive to promote and improve the synthetic of the framework materials such as the interior albumen of body, fat, obviously can improve the body condition of milk cow.

2.2 impacts of glucose precursor on milk cow energy balance

Table 4 Diseases of Cow blood plasma part metabolite testing result

The energy balance of milk cow can be evaluated by measuring blood plasma associated metabolic product.Result of the test shows, add glucose precursor and can obviously improve the concentration of glucose in Diseases of Cow blood plasma, reduce the concentration of free fatty and beta-hydroxybutyric acid, show that the satisfaction degree of glucose in Contents in Cows improves, the degree of decomposition of body fat reduces, and it is less that milk cow is employed body fat.The concentration of Postpartum Cows blood plasma free fatty acid and beta-hydroxybutyric acid is lower than the critical value (free fatty 500 μ Eq/L, beta-hydroxybutyric acid 1200 μ mol/mL) of clinical ketoacidosis diagnosis simultaneously.Use the nutrition regulation additive of Diseases of Cow special use can prevent the generation of milk cow negative energy balance.

2.3 impacts of glucose precursor on milk production of cow

Table 5120 day milk production of cow

?	Number (head)	Give milk number of days (my god)	Total output of milk (kg)	The average output of milk (kg/ days. head)
					Control group	23	120	105898	38.37±5.16
Test group	23	120	123703	44.82±5.92
					Process difference ^b	?	?	?	6.45

Above result shows: test group is compared with control group, and head day increases by 6.45 kilograms of the outputs of milk, improves 16.81%, t check significant difference (P < 0.05), and glucose precursor can significantly improve the milk yield of lactating cow.

2.4 impacts of glucose precursor on immunity of cow

Table 6 Diseases of Cow Immunoglobulins in Serum testing result

In cow serum, the content of Immunoglobulin IgA, IgM, IgG is the important indicator of judgement milk cow humoral immunity level.As seen from the experiment, cow birth stress cause serum immune globulin IgA, IgM, IgG content sharply to decline, and humoral immune function is suppressed.Before using glucose precursor to increase cow birth in Diseases of Cow daily ration, the content of 1 week, the childbirth same day and minute 1 week puerperium serum IgA, IgM, IgG, strengthens humoral immune function.

2.5 impacts of glucose precursor on Diseases of Cow reproductive performance

Table 7 Diseases of Cow reproductive performance testing result

Result of the test shows, Diseases of Cow is used glucose precursor can greatly shorten nearly 10 days of the time of milk cow postpartum heat, improve the conception rate 16.2% of the first feelings phase of postpartum milk cow, increase the ratio of oestrusing 16% of Postpartum Cows colony, effectively improve the reproductive performance of milk cow.

The impact that 2.6 glucose precursors occur Diseases of Cow postpartum disease

Table 8 Diseases of Cow postpartum disease testing result

Above result shows: test group is compared with control group, aspect prevention postpartum disease of cow, can significantly reduce the incidence of disease of various diseases, the average 13.0-30.5% that reduces, t checks significant difference (P < 0.05), glucose precursor can significantly reduce the incidence of the diseases such as milk cow mammitis in postpartum, endometritis, ketoacidosis, cud displacement, intractable apositia, puerperal fever and retention of afterbirth, improves the health status of milk cow.

Claims

1. a preparation method who improves cow rumen flora glucose precursor, it is characterized in that, comprise the steps: accurately to take microorganism formulation 45-55 part, propane diols 22-28 part, glycerine 22-28 part, calcium propionate 22-28 part, alumino-silicate 22-28 part, Chinese herbal medicine extract 18-22 part, niacinamide 12-18 part, nicotinic acid 12-18 part, sodium lactate 10-15 part, sodium chloride 10-15 part, complex enzyme 8-12 part, citric acid 5-10 part, rumen glucose 5-10 part, rumen-bypass amino acid 3-8 part, cross cud choline 5-8 part, B B-complex 3-5 part, composite mineral matter 2-4 part, first add respectively its quality 0.5-1 sterilized water doubly to dilute propane diols and glycerine, then dilution is mixed, heating water bath is to 30-50 ℃, and limit is uniformly mixed to obtain mixture A, heat preservation for standby use, calcium propionate, alumino-silicate, sodium lactate, sodium chloride, citric acid are evenly mixed to get to mixture B simultaneously, Chinese herbal medicine extract, rumen glucose, rumen-bypass amino acid, mistake cud choline, composite mineral matter are evenly mixed to get to mixture C, microorganism formulation, niacinamide, nicotinic acid, complex enzyme, B B-complex are evenly mixed to get to mixture D, then mixture B is added to LDH-1 type coulter type mixer, start mixer, then add mixture A evenly to mix 10-20min, then add mixture C, evenly mix 5-10min, finally add mixture D, evenly mix 3-5min, final mixture is sealed, packs and obtain the glucose precursor that can improve cow rumen flora,

Described microorganism formulation is occupy cud clostridium (Clostridium ruminocolum) CGMCC NO.2125 and aspergillus niger (Aspergillus niger) CGMCC NO.7927 through mixed culture fermentation, low-temperature negative-pressure Vacuum Concentration, fluidized bed drying, low-temperature grinding and make.

2. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, it is characterized in that, the preparation method of described microorganism formulation comprises the steps: first will occupy cud clostridium CGMCC NO.2125 and the activation of aspergillus niger CGMCC NO.7927 inclined-plane bacterial strain, expands and is cultured to first class seed pot respectively through one-level, secondary, three grades of seeds; Then by aspergillus niger CGMCCNO.7927 with occupy cud clostridium CGMCC NO.2125 first class seed pot liquid seeds by volume 1:0.5-1 evenly mix, with 10% inoculum concentration access secondary seed tank alternate, cultivate, alternate culture medium loading amount 240L, cultivation temperature 33-39 ℃, mixing speed 200-700r/m, ventilation (V/V) 1:1-3, incubation time 15-24h; Last secondary seed tank alternate liquid seeds is with 6% inoculum concentration access fermentation tank, cultivation temperature 28-30 ℃, mixing speed 200-700r/m, ventilation (V/V) 1:1-3, incubation time 15-20h; Then with 1-2 ℃/h rate of temperature fall slow cooling to 10-15 ℃, constant temperature culture 10-12h; Continuation to 2-5 ℃, now, is appended access fermentation tank, constant temperature culture 6-8h by secondary seed tank alternate liquid seeds with 4% inoculum concentration with 1-2 ℃/h rate of temperature fall slow cooling; Finally with 1-2 ℃/h heating rate, be slowly warming up to 10-15 ℃, constant temperature culture 10-12h; Continuation is slowly warming up to 37-39 ℃, constant temperature culture 15-20h with 1-2 ℃/h heating rate; Zymotic fluid is concentrated in vacuo to 45% of original volume through low-temperature negative-pressure, obtains bacterium concentrate, and concentrate evenly mixes for 0.5:1 in mass ratio with carrier, in 45 ℃ of fluidized bed dryings, and 40 ℃ of low-temperature grinding, grinding particle size 0.5-1.5mm, obtains microorganism formulation.

3. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 2, is characterized in that, described vehicle weight umber consists of: CaCO ₃20-23 part, dextrin 10-12 part.

4. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1 or 2, is characterized in that, described microorganism formulation viable count content is 1.5x10 ¹⁰-3x10 ¹⁰individual/g.

5. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, is characterized in that, described alumino-silicate is alumino-silicate or alumino-silicate derivatives.

6. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, it is characterized in that, the preparation method of described Chinese herbal medicine extract is: accurately take Radix Angelicae Sinensis 55-65 part, Ligusticum wallichii 55-65 part, fructus amomi 50-60 part, radix paeoniae rubrathe 50-60 part, cultivated land 50-60 part, Divine Comedy 45-55 part, Fructus Hordei Germinatus 45-55 part, motherwort 45-55 part, costus root 40-45 part, Radix Codonopsis 40-45 part, rhizoma atractylodis 40-45 part, Radix Astragali 40-45 part, honeysuckle 40-45 part, cordate houttuynia 35-45 part, Poria cocos 35-45 part, jujube kernel 35-45 part, fushen 35-45 part, polygala root 30-40 part, fry fennel 30-40 part, bighead atractylodes rhizome 30-40 part, bark of official magnolia 30-40 part, Fructus Aurantii 25-35 part, Radix Glycyrrhizae 25-35 part, hawthorn 25-35 part, dried orange peel 25-35 part, respectively said herbal medicine being crushed to particle diameter is below 2 millimeters, then in container, evenly mix and add the water of 3-6 times of weight, control temperature 70 C-90 ℃ and keep 2-4h, then be cooled to 45-60 ℃, add the mixing enzyme preparation of mixed material gross weight 5-10% to carry out enzymolysis, with newborn acid for adjusting pH value, be 5.5-6.8, enzymolysis 2-4h, finally add 75% ethanol of mixed material 0.5-3 times weight and the mixture of propyl alcohol, the mass ratio that described ethanol and propyl alcohol mix is 1:1-2, control temperature to 60 ℃-78 ℃ of maintenance 3-4h, filter, it is more than 20% that filtrate decompression is concentrated into solid content, and then freeze drying, 40 ℃ of low-temperature grinding obtain the Chinese herbal medicine extract that grinding particle size is 0.5-2mm.

7. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 6, it is characterized in that, the parts by weight of described mixed enzyme consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, beta-glucosidase 10-15 part, zytase 15-20 part, pentosanase 15-20 part, Pullulanase 20-30 part, beta amylase 10-15 part, neutral proteinase 10-15 part, acid protease 10-15 part, superoxide dismutase 5-10 part, glucose oxidase 5-10 part, acid phosphatase 5-10 part.

8. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, it is characterized in that, described complex enzyme is evenly mixed by the solid enzyme preparation of following parts by weight: neutral proteinase 22-28 part, acid protease 22-28 part, alkalescent xylanase 22-28 part, acidic xylanase 22-28 part, cellulase 22-28 part, 1,4 beta-glucanase 15-20 part, pectase 15-20 part, amylase 15-20 part, seminase 10-15 part, phytase 10-15 part.

9. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, is characterized in that, described per kilogram B B-complex comprises dehydroretinol 800mg, orotic acid 000mg, Catergen 000mg, riboflavin 1600mg, pantothenic acid 1900mg, pyridoxamine 1800mg, cobalamin 1200mg, inositol 5000mg, vitamin E 600mg, neo dohyfral D3 5500mg, prokayvit 4000mg; Its surplus is carrier, and described carrier is rice bran.

10. the preparation method who improves cow rumen flora glucose precursor as claimed in claim 1, it is characterized in that, described per kilogram composite mineral matter comprises calcium 6000mg, phosphorus 4200mg, sodium 5100mg, copper 800mg, iron 1800mg, potassium 3800mg, manganese 1400mg, cobalt 1600mg, iodine 5000mg, magnesium 1450mg; Its surplus is carrier, and described carrier is rice bran.