CN103689218B - A kind ofly improve the composite bacteria agent capable and application thereof that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate - Google Patents
A kind ofly improve the composite bacteria agent capable and application thereof that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate Download PDFInfo
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- CN103689218B CN103689218B CN201310653964.1A CN201310653964A CN103689218B CN 103689218 B CN103689218 B CN 103689218B CN 201310653964 A CN201310653964 A CN 201310653964A CN 103689218 B CN103689218 B CN 103689218B
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Abstract
The invention discloses belong to animal and fowl fodder technical field a kind of and improve the composite bacteria agent capable and application thereof that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate.Described composite bacteria agent capable is made up of bacillus subtilis, candida utili and VREF.Composite bacteria agent capable provided by the invention, can significantly improve rumen microorganism dietary fiber digestibility, and increase rumen microbial fermentation general volatile aliphatic acid growing amount up to 21.1%, gluconeogenesis precursor propionic acid growing amount positive regulation is up to 10.5 ~ 11.3%.Preparation method is simple, workable, is applicable to livestock and poultry breeding industry feed manufacturing, significantly can promote ruminant Energy utilization ratio.
Description
Technical field
The invention belongs to animal and fowl fodder technical field, particularly a kind ofly improve the composite bacteria agent capable and application thereof that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate.
Background technology
The carbohydrate such as fiber, starch and soluble sugar in searched for food feed can be converted into pyruvic acid through digestion and metabolism by ruminant domestic animal rumen microorganism usually, enter tricarboxylic acid cycle and generate the volatile fatty acids (VFA) such as acetic acid, propionic acid and butyric acid respectively by different metabolic pathways.Concerning lactating cow, acetic acid is important as precursors thing in mammary gland synthesis butterfat.Acetic acid also may be used for synthesis a small amount of glucose and ketoboidies in vivo, and most of acetic acid is in vivo mainly through acetyl coenzyme A compound body fat and butterfat under reducibility coenzyme II effect, and storage power in this way; Acetic acid also can enter tricarboxylic acid cycle in vivo and resolve into CO
2and H
2, discharge ATP energy supply simultaneously.The major way that ruminant digestion absorbs carbohydrate is VFA, instead of glucose, for keeping body blood sugar concentration, the gluconeogenesis precursor propionic acid that lumen fermentation generates plays very important effect, in addition, propionic acid is not only lactating cow and is improved the important as precursors thing of lactose, and provides in energy and the raising output of milk at body and have important effect.Butyric acid is first converted into beta-hydroxy-butanoic acid in cud wall absorption process and is entered liver and breast tissue, participates in the metabolism of substance in vivo, especially as the energy source of musculature.Therefore, acetic acid, propionic acid and butyric acid are indispensable metabolites to lactating cow.It is generally acknowledged that in cud chyme liquid, total VFA generation is directly related with feed digestibility, acetic acid, propionic acid and butyric acid proportion of composing are then main relevant with Non-Structural Carbohydrate ratio with structural carbohydrate in feed.
Have Research Literature to report, in feed, single interpolation yeast microorganism fodder can reduce lumen fermentation partial pressure of oxygen, prevents lactic acid from excessively generating, and improves rumen digestion; During the microorganism fodders such as single interpolation such as lactic acid bacteria, enterococcus, bacillus, Propionibacterium, Megasphaera elsdenii, contestable suppresses the growth of pathogen, immune stimulatory function, regulates microbial balance in intestines and stomach; Some lactic acid bacterias can control diarrhea disease percentage, improve body weight gains and feed conversion rate, and lactic acid utilizes bacterium then can improve calf body weight gains, have the effect improving children calf in age health and upgrowth situation.But some microorganism fodder above-mentioned during single interpolation, although can improve digestibility in various degree, cannot realize the generation ratio (NGR) of directed regulation and control lumen fermentation non-gluconeogenesis precursor acetic acid and butyric acid and gluconeogenesis precursor propionic acid in feed.
Summary of the invention
The object of the present invention is to provide and a kind ofly improve the composite bacteria agent capable and application thereof that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate, described composite bacteria agent capable can improve feed fibre degradation rate and positive regulation lumen fermentation gluconeogenesis precursor propionic acid generates.
Improve the composite bacteria agent capable that feed fibre degradation rate and positive regulation rumen fermentation propionic acid generate, this composite bacteria agent capable is made up of bacillus subtilis (Bacillussubtilis), candida utili (Candidautilis) and VREF (Enterococcusfaecium); Described bacillus subtilis (Bacillussubtilis) its viable count is not less than 2 × 10
10cfu/ gram of dry weight; Described candida utili (Candidautilis) its viable count is not less than 4 × 10
9cfu/ gram of dry weight, described VREF (Enterococcusfaecium) its viable count is not less than 1 × 10
10cfu/ gram of dry weight.
Described bacillus subtilis (Bacillussubtilis), candida utili (Candidautilis) are (1 ~ 3) with VREF (Enterococcusfaecium) bacterium powder according to weight ratio: (1 ~ 3): the proportion compatibility of (1 ~ 3) mixes.
The application of above-mentioned composite bacteria agent capable in domestic animal feed manufacturing.
The described application be applied as in raising dietary fiber degradation rate and positive regulation lumen fermentation gluconeogenesis precursor propionic acid generate.
Described application is following either method:
1) lactating cow TRM: mix by adding composite bacteria agent capable 1000g in TRM per ton;
2) cattle and sheep concentrate supplement is fattened: mix according to adding composite bacteria agent capable 1000 ~ 2000g in concentrate supplement per ton.
Beneficial effect of the present invention: and composite bacteria agent capable provided by the present invention, different roughage ratio diet dry digestibility can be significantly improved up to 6.4%, improve fiber testing 2.2 ~ 45.7%, significantly improve rumen microbial fermentation general volatile aliphatic acid growing amount 4.6 ~ 21.1%, directed regulation and control rumen microbial fermentation gluconeogenesis precursor propionic acid generates ratio and improves 10.5 ~ 11.3%, has and promote effect significantly in promotion ruminant Energy utilization ratio.
Detailed description of the invention
The present invention is set forth further below in conjunction with specific embodiment.Unless specifically stated otherwise, experimental technique used in the present invention and technology are Method and Technology known in those skilled in the art.
To those skilled in the art, under the prerequisite not deviating from the present invention's spirit and enforcement, the various change carry out the methane phase inhibitor component, content, ratio etc. in these embodiments or change also belong to protection scope of the present invention.
The carbohydrate such as fiber, starch and soluble sugar in searched for food diet can be converted into pyruvic acid through a series of digestion and metabolism approach by ruminant domestic animal rumen microorganism protozoon, bacterium, fungi and ancient bacterium usually, enter tricarboxylic acid cycle and generate the volatile fatty acids (VFA) such as acetic acid, propionic acid and butyric acid respectively by different metabolic pathways.Diet digestibility and rumen microbial fermentation VFA growing amount and ratio thereof not only by add the impact of microorganism fodder but also with roughage ratio in diet, there is substantial connection, be below that the embodiment made the effect of composite bacteria agent capable in the present invention is as follows under different diet roughage ratio condition:
Wherein bacillus subtilis used (Bacillussubtilis) (hereinafter referred to as BS) microbial inoculum and candida utili (Candidautilis) (hereinafter referred to as CU) microbial inoculum are that Cangzhou, Hebei prosperous generation thing technical research institute produces, and VREF (Enterococcusfaecium) (hereinafter referred to as EF) microbial inoculum is that Zhengzhou lilium auratum lindle bioengineering Co., Ltd produces.
Embodiment 1
The analyzing of applying effects that under high coarse fodder ratio condition, composite bacteria agent capable generates diet rumen microorganism fiber degradation and gluconeogenesis precursor
By bacillus subtilis (Bacillussubtilis) (hereinafter referred to as BS), candida utili (Candidautilis) (hereinafter referred to as CU) and VREF (Enterococcusfaecium) (hereinafter referred to as EF) three kinds of microorganism fodders, employing L
9(3
3) orthogonal arrage (table 1) carried out analytical test for these three microorganism fodder compatibility addition combined effects and compared.Bacillus subtilis used (Bacillussubtilis) microbial inoculum, be light yellow solid powder, moisture is not higher than 8%, and viable count is not less than 2 × 10
10cfu/ gram of dry weight; Candida utili (Candidautilis) microbial inoculum, be white solid powder, moisture is not higher than 10%, and viable count is not less than 4 × 10
9cfu/ gram of dry weight, VREF (Enterococcusfaecium), be white solid powder, moisture is not higher than 8%, and viable count is not less than 1 × 10
10cfu/ gram of dry weight.
Table 1 microorganism fodder addition compatibility combination orthogonal test L
9(3
3) table
In table, BS: bacillus subtilis, CU: candida utili, EF: VREF
With the donor animal that lactation period, holstein cow was rumen fluid in this test, by sheep's hay and fine fodder, (every kilogram of fine fodder contains: corn flour 404.7 grams, dregs of beans 176.6 grams, rapeseed dregs 65.5 grams, Cottonseed Meal 33.3 grams, expanded soybean 32.2 grams, megasse 82.9 grams, cottonseed 163.3 grams, stone flour 11.6 grams, salt 7.2 grams, 14.4 grams, sodium bicarbonate, vitamin and microelement pre-mix material 8.3 grams) mix in 70:30 ratio, be mixed with high coarse fodder ratio TRM as diet substrate, take 0.5 gram of diet substrate in anaerobic fermentation bottle with assay balance.In anaerobic fermentation bottle, the corresponding bacillus subtilis of each compatibility combined treatment group (Bacillussubtilis) is added according to table 1, candida utili (Candidautilis) and VREF (Enterococcusfaecium), add the liquid anaerobic fermentation medium (culture medium composition is in table 2) of 50mLpH6.85, inoculation 25mL lactating cow rumen fluid is (wherein containing cud chyme mixed microorganism, comprise endogenous bacteria, fungi, ancient bacterium and protozoon etc.), carry out anaerobic fermentation after 72 hours, measure the external dry disappearance rate (IVDMD) of diet substrate, substrate neutral detergent fiber degradation rate (NDFD), acetic acid in acid detergent fiber degradation rate (ADFD) and zymotic fluid in substrate, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid concentration, and calculate general volatile acid concentration (tVFA).Calculate the sugared volatile fatty acid of non-life and gluconeogenesis volatile fatty acid ratio: NGR=(acetic acid+2 × butyric acid+valeric acid)/(propionic acid+valeric acid).Whole test, each compatibility combination arrangement 4 fermentation flasks repeat, and repeat three batch fermentation.
Table 2, fluid nutrient medium form
| Solution component | Liquor capacity, mL |
| Solution A | 200 |
| B solution | 200 |
| C solution | 200 |
| D reducing solution | 40 |
| E resazurin solution | 1.0 |
| Distilled water | 400 |
In table, solution A is trace element solution, and often liter containing 132 grams of CaCl
22H
2o, 100 grams of MnCl
24H
2o, 10 grams of CoCl
26H
2o, 80 grams of FeCl
36H
2o; B solution is carbonate buffer solution, and often liter containing 4 grams of NH
4hCO
3, 35 grams of NaHCO
3; C solution is PBS, and often liter containing 5.7 grams of Na
2hPO
4, 6.2 grams of KH
2pO
4, 0.6 gram of MgSO
47H
2o; D reducing solution often rises containing 1.6 grams of NaOH, 6.25 grams of Na
2s9H
2o.E resazurin solution, often liter containing 1 gram of resazurin indicator.
Test result under table 3, high roughage ratio diet condition
In table, IVDMD: external dry disappearance rate; NDFD: neutral detergent fiber degradation rate; ADFD: acid detergent fiber degradation rate; TVFA: total volatile acid growing amount; ACE: acetic acid proportion; PRO: propionic acid proportion; BUT: butyric acid proportion; NGR: the ratio of the sugared volatile acid of non-life and gluconeogenesis volatile acid.
Table 3 result of the test shows, under high roughage ratio diet condition, after different microorganism fodder compatibility combined treatment group rumen microbial fermentation 72h, IVDMD all has rising in various degree, wherein, promote effect in compatibility combined treatment group 7 and 9 the most remarkable, improve 6.0% compared to control group; Different microorganism fodder compatibility combined treatment can improve the NDFD of each processed group in various degree, promote effect the most remarkable, improve 10.9% compared to control group in processed group 5; Different microorganism fodder compatibility combined treatment can improve total volatile acid growing amount of each processed group in various degree, promote effect the most remarkable, improve 21.1% compared to control group in processed group 3; After adding composite bacteria agent capable feed addictive, in total volatile acid, acetic acid proportion reduces in various degree, reduces effect the most remarkable, reduce 4.3% compared to control group in processed group 3 and 9; Propionic acid proportion then raises in various degree with the use of composite bacteria agent capable feed addictive, promotes effect the most remarkable, improve 10.5% compared to control group in processed group 3.Butyric acid proportion also promotes in various degree after interpolation composite bacteria agent capable feed addictive, wherein promotes effect in processed group 2,3 and 5 the most remarkable, improves 4.1% compared to control group.NGR level then reduces in various degree because adding composite bacteria agent capable feed addictive, when substrate is C3F7, reduces effect the most remarkable, reduce 15.3% compared to control group in processed group 3.
Embodiment 2: the analyzing of applying effects that under low coarse fodder ratio condition, composite bacteria agent capable generates diet rumen microorganism fiber degradation and gluconeogenesis precursor
With lactation period holstein cow for experimental animal, by sheep's hay and fine fodder, (every kilogram of fine fodder contains: corn flour 404.7 grams, dregs of beans 176.6 grams, rapeseed dregs 65.5 grams, Cottonseed Meal 33.3 grams, expanded soybean 32.2 grams, megasse 82.9 grams, cottonseed 163.3 grams, stone flour 11.6 grams, salt 7.2 grams, 14.4 grams, sodium bicarbonate, premix 8.3 grams) become low coarse fodder ratio TRM as diet substrate in 30:70 proportions, take 0.5 gram of diet substrate in anaerobic fermentation bottle with assay balance.According to table 1, the corresponding bacillus subtilis of each compatibility combined treatment group is added in anaerobic fermentation bottle, candida utili and VREF, add the liquid anaerobic fermentation medium (culture medium composition is see table 2 in embodiment 1 Suo Shi) of 50mLpH6.85, inoculation 25mL lactating cow cud chyme mixed microorganism, carry out anaerobic fermentation after 72 hours, measure the external dry disappearance rate (IVDMD) of feed substrate, substrate neutral detergent fiber degradation rate (NDFD), acetic acid in acid detergent fiber degradation rate (ADFD) and zymotic fluid in substrate, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid concentration, and calculate general volatile acid concentration (tVFA).Calculate the sugared volatile fatty acid of non-life and gluconeogenesis volatile fatty acid ratio: NGR=(acetic acid+2 × butyric acid+valeric acid)/(propionic acid+valeric acid).Whole test, each compatibility combination arrangement 4 fermentation flasks repeat, and repeat three batch fermentation.
Test result under table 4, low roughage ratio diet condition
In table, IVDMD: external dry disappearance rate; NDFD: neutral detergent fiber degradation rate; ADFD: acid detergent fiber degradation rate; TVFA: total volatile acid growing amount; ACE: acetic acid proportion; PRO: propionic acid proportion; BUT: butyric acid proportion; NGR: the ratio of the sugared volatile acid of non-life and gluconeogenesis volatile acid.
Table 4 result of the test shows under low roughage ratio diet condition, after different microorganism fodder compatibility combined treatment group rumen microbial fermentation 72h, IVDMD all has rising in various degree, promote effect in processed group 9 the most remarkable, improve 6.4% compared to control group.Different microorganism fodder compatibility combined treatment can improve the NDFD of each processed group in various degree, promote effect the most remarkable, improve 15.5% compared to control group in processed group 9.Add the ADFD that composite bacteria agent capable feed addictive can improve each processed group in various degree, promote effect in processed group 9 the most remarkable, improve 45.7% compared to control group.Different microorganism fodder compatibility combined treatment can improve total volatile acid growing amount of each processed group in various degree, promote effect the most remarkable, improve 20.7% compared to control group in processed group 9.After adding composite bacteria agent capable feed addictive, in total volatile acid, acetic acid proportion reduces in various degree, reduces effect the most remarkable, reduce 3.2% compared to control group in processed group 9; Propionic acid ratio then raises in various degree with the use of composite bacteria agent capable feed addictive, promotes effect the most remarkable, improve 11.3% compared to control group in processed group 9.Butyric acid ratio also promotes in various degree after interpolation composite bacteria agent capable feed addictive, wherein promotes effect in processed group 5 the most remarkable, improves 8.1% compared to control group.NGR level reduces in various degree because adding composite bacteria agent capable feed addictive, reduces effect the most remarkable, reduce 13.2% compared to control group in processed group 9.
Comprehensive above-described embodiment 1 and 2, test result shows, bacillus subtilis in composite bacteria agent capable composite bacteria agent capable of the present invention, candida utili and VREF are (1 ~ 3) at addition proportion compatibility: (1 ~ 3): time (1 ~ 3), different roughage ratio diet dry digestibility can be significantly improved up to 6.4%, improve fiber testing 2.2 ~ 45.7%, significantly improve rumen microbial fermentation general volatile aliphatic acid growing amount 4.6 ~ 21.1%, directed regulation and control rumen microbial fermentation gluconeogenesis precursor propionic acid generates ratio and improves 10.5 ~ 11.3%, have in promotion ruminant Energy utilization ratio and promote effect significantly.
Claims (4)
1. the composite bacteria agent capable improving feed fibre degradation rate and positive regulation rumen fermentation propionic acid and generate, it is characterized in that, this composite bacteria agent capable is made up of bacillus subtilis (Bacillussubtilis), candida utili (Candidautilis) and VREF (Enterococcusfaecium); Described bacillus subtilis (Bacillussubtilis) its viable count is not less than 2 × 10
10cfu/ gram of dry weight; Described candida utili (Candidautilis) its viable count is not less than 4 × 10
9cfu/ gram of dry weight, described VREF (Enterococcusfaecium) its viable count is not less than 1 × 10
10cfu/ gram of dry weight;
Wherein bacillus subtilis (Bacillussubtilis), candida utili (Candidautilis) are (1 ~ 3) with VREF (Enterococcusfaecium) bacterium powder according to weight ratio: (1 ~ 3): the proportion compatibility of (1 ~ 3) mixes.
2. the application of composite bacteria agent capable described in claim 1 in domestic animal feed manufacturing.
3. application according to claim 2, is characterized in that, described in be applied as improve dietary fiber degradation rate and positive regulation lumen fermentation gluconeogenesis precursor propionic acid generate in application.
4. application according to claim 2, is characterized in that, is following either method:
1) lactating cow TRM: mix by adding composite bacteria agent capable 1000g in TRM per ton;
2) cattle and sheep concentrate supplement is fattened: mix according to adding composite bacteria agent capable 1000 ~ 2000g in concentrate supplement per ton.
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| CN105981915A (en) * | 2015-02-03 | 2016-10-05 | 中国科学院亚热带农业生态研究所 | Application of active saccharomycetes in improving utilization efficiency of coarse feed |
| CN104862258A (en) * | 2015-06-11 | 2015-08-26 | 中国科学院成都生物研究所 | Complex microbial agent for high-organic-load and high-temperature biogas fermentation as well as preparation method and application thereof |
| CN105192250B (en) * | 2015-10-20 | 2019-07-12 | 中国农业科学院农业资源与农业区划研究所 | A kind of feeding type probiotics and its application |
| CN105647830B (en) * | 2016-02-29 | 2019-05-14 | 海南泓缘生物科技股份有限公司 | It is a kind of to enhance the composite bacteria agent of animal immunizing power, preparation method and applications |
| CN111838427A (en) * | 2020-08-11 | 2020-10-30 | 南京农业大学 | Phytosterol formula for promoting rumen fermentation and determination method thereof |
| CN113008822B (en) * | 2021-03-10 | 2022-04-22 | 江西农业大学 | Method for measuring degradation rate of pyruvic acid creatine in rumen fluid of ruminant |
| CN114271387A (en) * | 2022-01-07 | 2022-04-05 | 河南农业大学 | Additive composition for promoting sheep feed fiber digestion and rumen health and preparation method and application thereof |
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