CN106173339A - The application of glyceryl trinitrate - Google Patents
The application of glyceryl trinitrate Download PDFInfo
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- CN106173339A CN106173339A CN201610575073.2A CN201610575073A CN106173339A CN 106173339 A CN106173339 A CN 106173339A CN 201610575073 A CN201610575073 A CN 201610575073A CN 106173339 A CN106173339 A CN 106173339A
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- methane
- glyceryl trinitrate
- methane backeria
- inhibitor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/22—Methane [CH4], e.g. from rice paddies
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Abstract
The invention discloses glyceryl trinitrate application in preparing methane backeria inhibitor.The invention also discloses the glyceryl trinitrate application as animal feed additive.Compared with prior art, the present invention can effectively reduce the generation of methane, and its long-lasting, safety and biological tolerance are all better than existing product.
Description
Technical field
The invention belongs to feed additive field, be specifically related to the application of glyceryl trinitrate.
Background technology
Cud methane generates and does not only result in the ruminant feed energy loss of 5～7%, or greenhouse gases important come
Source, 10～16kg methane, cattle average out to 60～160kg are produced in the every annual of sheep.Therefore, the research to ruminant methane inhibitor
Application, can not only improve energy utilization efficiency and the production performance of animal, also have good ecological benefits.Although it is existing various
The feed additive (such as natural plant extracts, ionophore, many halogen compounds, electron acceptor, oils and fats etc.) of various kinds is ground
Study carefully exploitation, but all there is different problems, such as problems such as long-lasting, safety and microbiological resistance.These drawbacks limit
The extensive application of existing additive.Nitrate is that Recent study is more, is also the cud methane suppression having wide application prospects
Agent, but its intermediate product nitrite of metabolism in cud, it is possible to enter blood through cud wall and cause methemoglobinemia
Disease, endangers animal health.Although the research of safety applications nitrate has been carried out, but still longer stretch is had to walk.Therefore, one
Planting new, efficient ruminant methane inhibitor is that society is needed badly.
Summary of the invention
The technical problem to be solved in the present invention is to provide glyceryl trinitrate application in preparing methane backeria inhibitor,
The problems such as long-lasting, the safety of solution prior art existence and microbiological resistance deficiency.
The present invention also to solve the technical problem that the application being to provide glyceryl trinitrate as animal feed additive.
For solving above-mentioned technical problem, the technical solution used in the present invention is as follows:
Glyceryl trinitrate application in preparing methane backeria inhibitor.
The structural formula of glyceryl trinitrate is as follows:
Wherein, described methane backeria is cud methane backeria.
Wherein, described methane backeria inhibitor is ruminant methane backeria inhibitor.
Glyceryl trinitrate as the application of animal feed additive also within protection scope of the present invention.
Wherein, described animal is ruminant.
Wherein, described application refer to glyceryl trinitrate as animal feed additive suppression animal rumens methane in
Application.
Beneficial effect:
Compared with prior art, the present invention has the advantage that
Glycerin trinitrate can be by the nitrate functional group (-O-NO of self2) oxidizing of methylene fungus beetle base coenzyme M reductase
Nickel (+1) active center.Methyl coenzyme M reductase is the key enzyme in methane backeria methane constructive ways, and its inactivation will block
Methane generates.Experiment proves that glycerin trinitrate has the ability that long-term suppression cud methane generates.
Compared with the cud methane inhibitor nitrate wider with research at present, its safety is higher, because glycerin trinitrate
During ruminal metabolism, it is not easily caused nitrite excess accumulation.In the experiment in vitro implemented and animal experiment in vivo all
Do not detect nitrite ion accumulation.Additionally, glycerin trinitrate is as the disease treatment medicine of mankind's life-time service, its pharmacology
Character is the clearest and the most definite, and animals and humans health will not cause unknown harm.
Detailed description of the invention
According to following embodiment, the present invention be may be better understood.But, as it will be easily appreciated by one skilled in the art that reality
Execute the content described by example and be merely to illustrate the present invention, and should be also without limitation on basis described in detail in claims
Invention.
Embodiment 1
1 MATERIALS METHODS
1.1 experiment material
1.1.1 experimental animal and rumen fluid collection
Experimental animal is 4 sheep equipped with permanence lymphoma stomach fistulization pipe.Feed alfalfa hay, freely drink water.Every day 08:00
Feed at twice with 16:00.Testing the same day, before raising morning, half an hour takes rumen fluid, obtains fresh rumen fluids from 4 Fistulated sheep fast
Quick-mounting enters to be passed through in advance the thermos flask of carbon dioxide and takes back laboratory.
1.1.2 prepared by artificial rumen fermentation liquid
The preparation of fermentation liquid is with reference to the method for Menke and Steingass (1988).By the fresh rumen fluid of collection with 4 layers
Filtered through gauze, measures in the artificial buffer that the rumen fluid of 1250mL pours rapidly off-the-shelf 3750mL into, and rumen fluid is with slow
Rush liquid to mix in the ratio of 1:3 (v/v), make mixing artificial rumen culture fluid.Mixed fermentation liquid is dispensed into 180mL send out
In ferment bottle, the temperature of whole process is maintained at 39 DEG C, is continually fed into pure carbon dioxide to keep anaerobic environment (to become with "diazoresorcinol"
Colourless judge), by magnetic stirrer to keep rumen fluid to mix homogeneously with buffer.Fermentation flask adds 0.5g in advance
The methane backeria inhibitor pulverizing alfalfa hay and 0.5g Semen Maydis powder and various dose is placed in the constant incubator of 39 DEG C preheating
30min.Then in fermentation flask, carbon dioxide it is passed through to ensure in fermentation flask as anaerobic environment.After subpackage, shake at constant temperature
Swing device 39 DEG C, cultivate 24 hours under the conditions of 100r/min.
1.1.3 methane backeria inhibitor
Glyceryl trinitrate (Nitroglycerin, NG)
1.1.4 prepared by anaerobic fungi culture medium
Culture medium preparation is with reference to the method preparation of (1993) such as Davies: by buffer, basal medium, acellular cud
Liquid, reducing agent and oxidation-reduction indicator composition.Substrate is rice straw, and diameter is about 1mm, and addition is 1% (w/v).
1.2 test method
1.2.1 EXPERIMENTAL DESIGN
Embodiment 1: test sets 3 groups, matched group (without any medicine), nitroglycerin group 1 (6.6 μm/L) and nitric acid
Glycerol group 2 (33 μm/L).
Embodiment 2: test sets 4 groups, matched group (without any medicine), nitroglycerin group 1 (6.6 μm/L), nitric acid
Glycerol group 2 (13.2 μm/L) and nitroglycerin group 3 (19.8 μm/L).
1.2.2 chemical analysis
During fermentation ends, measure total gas production, H immediately2And CH4And pH.Take 2mL fermentation liquid, 13 000r/min and 4 DEG C of bars
Under part, centrifugal 10min takes 1.5mL supernatant, adds 0.2mL 25% Metaphosphoric acid and fixes, after standing 15min, and-20 DEG C of preservations, after
Continue the mensuration for volatile fatty acid.
The mensuration of total gas production, with reference to the method for (1994) such as Theodorou, is produced in measuring fermentation flask with air pressure conversion device
Tolerance.
H2And CH4Use Agilent 7890B gas chromatograph for determination.Temperature of vaporization chamber: 200 DEG C；Chromatographic column (Porapak Q
Packing&MolSieve 5A packing, Agilent), column temperature 80 DEG C；TCD detector: 200 DEG C；Carrier gas is N2。
Volatile fatty acid is with reference to Pontes etc.[13]Method detects.Use Agilent 7890B gas chromatograph.Vaporizer
220℃；Chromatographic column is FUSED SILICA capillary column (Supelco), column temperature 40 DEG C, 4min；40-110(40℃/min)；
110 DEG C, 2min；110-150(40℃/min)；150 DEG C, 5min gradient increased temperature.Hydrion flame detector FID:250 DEG C.Carry
Gas is nitrogen.Sample is acidified through Metaphosphoric acid, is inside designated as .beta.-methylacrylic acid.
Lactic acid measures with reference to Barker&Summerson (1941) method.
Dry disappearance rate measures with reference to (2001) methods such as Zhu Weiyun.
1.2.3 microorganism concn measures
Microorganism concn quantitative analysis usesPremix Ex TagTM (TaKaRa) reagent, Applied
Biosystems 7300Real-Time PCR System.The plasmid containing genes of interest fragment is utilized to prepare standard curve, meter
Calculate genes of interest concentration in the sample.PCR primer used herein is listed below.
PCR primer used herein
1.3 data statistics processing
One-way ANOVA (Tukey) in data acquisition SPSS 20.0 software is analyzed, and confidence interval is 95%.
From table 1, compared with matched group (being not added with group), the glycerin trinitrate adding 6.6 μm/L can reduce
The methane-generated quantity of 33.2%, the glycerin trinitrate adding 33.0 μm/L can completely inhibit methane generation.To short-chain fatty acid
Detection show (table 2), adding glycerin trinitrate and change the composition of short-chain fatty acid, acetic acid concentration reduces, propanoic acid and butanoic acid
Concentration raises, but the concentration of total short-chain fatty acid does not has significant change.
Table 1 ferment 24 hours gas generated
Table 2 ferments 24 hours short-chain fatty acid growing amounts
As shown in Table 3, the nitroglycerin adding 33.0 μm/L significantly reduces methane backeria quantity, but has no significant effect tumor
The quantity of the quantity of stomach anaerobic fungi, rumen metabolism and antibacterial.In cud, antibacterial and protozoon are the Main Bacterias of degraded feedstuff
Group, and anaerobic fungi is one of flora growing plant fragments the most surely, they rise in cud plant cell wall degradation process
Important function.Result of the test shows that nitroglycerin has specificity to the suppression of cud methane backeria.
Table 3 adds the glycerin trinitrate impact on cud methane backeria, antibacterial, fungus and protozoon
Embodiment 2 glycerin trinitrate directly acts on the checking test of methane backeria
Test uses the coculture checking of anaerobic fungi and methane backeria.From table 4, the glycerin trinitrate of 6.6 μm/L
The methane completely inhibiting coculture generates, and have accumulated substantial amounts of hydrogen simultaneously.A large amount of accumulation hint nitroglycerin of hydrogen do not have
There is the activity of suppression anaerobic fungi, but directly act on methane backeria.After methane backeria activity is suppressed, hydrogen is caused to amass in a large number
Tired, the hydrogen gas accumulation feedback suppression energy metabolism path of anaerobic fungi, cause total gas production and the reduction of disappearance of substrate rate.Nitre
Acid glyceride directly acts on methane backeria, suppresses it to generate the activity of methane, and this screens target with the design of inhibitor and is consistent.With
The increase of nitroglycerin dosage, total gas production, disappearance of substrate rate and hydrogen accumulation all significantly reduce, the nitre that hint is heavy dose of
Acid glyceride can suppress the activity of anaerobic fungi.This result requires that the use of glycerin trinitrate strictly to control dosage.
Table 4 nitroglycerin is on anaerobic fungi and the impact of methane backeria co culture system in vitro Rice Straw Fermented
Claims (6)
1. glyceryl trinitrate application in preparing methane backeria inhibitor.
Application the most according to claim 1, it is characterised in that described methane backeria is cud methane backeria.
Application the most according to claim 1, it is characterised in that described methane backeria inhibitor is that ruminant methane backeria presses down
Preparation.
4. glyceryl trinitrate is as the application of animal feed additive.
Application the most according to claim 4, it is characterised in that described animal is ruminant.
Application the most according to claim 4, it is characterised in that described application refers to that glyceryl trinitrate is as animal feed
Additive application in suppression animal rumens methane.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114052131A (en) * | 2021-12-08 | 2022-02-18 | 南京农业大学 | Application of glycerol trinitrate and disodium fumarate to preparation of rumen methane inhibitor |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102781252A (en) * | 2009-12-11 | 2012-11-14 | 帝斯曼知识产权资产管理有限公司 | Nitrooxy alkanoic acids and derivatives thereof in feed for reducing methane emission in ruminants, and/or to improve ruminant performance |
| CN103260424A (en) * | 2010-12-20 | 2013-08-21 | 帝斯曼知识产权资产管理有限公司 | Use of nitrooxy organic molecules in feed for reducing methane emission in ruminants, and/or to improve ruminant performance |
| CN103547168A (en) * | 2011-05-23 | 2014-01-29 | 格拉斯普工商业有限公司 | Use of encapsulated nitrates and sulfates to reduce methane emission derived from ruminal fermentation |
| CN103561586A (en) * | 2011-05-26 | 2014-02-05 | 帝斯曼知识产权资产管理有限公司 | Use of a feed composition for reducing methane emission in ruminants, and/or to improve ruminant performance |
-
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- 2016-07-20 CN CN201610575073.2A patent/CN106173339A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102781252A (en) * | 2009-12-11 | 2012-11-14 | 帝斯曼知识产权资产管理有限公司 | Nitrooxy alkanoic acids and derivatives thereof in feed for reducing methane emission in ruminants, and/or to improve ruminant performance |
| CN103260424A (en) * | 2010-12-20 | 2013-08-21 | 帝斯曼知识产权资产管理有限公司 | Use of nitrooxy organic molecules in feed for reducing methane emission in ruminants, and/or to improve ruminant performance |
| CN103547168A (en) * | 2011-05-23 | 2014-01-29 | 格拉斯普工商业有限公司 | Use of encapsulated nitrates and sulfates to reduce methane emission derived from ruminal fermentation |
| CN103561586A (en) * | 2011-05-26 | 2014-02-05 | 帝斯曼知识产权资产管理有限公司 | Use of a feed composition for reducing methane emission in ruminants, and/or to improve ruminant performance |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114052131A (en) * | 2021-12-08 | 2022-02-18 | 南京农业大学 | Application of glycerol trinitrate and disodium fumarate to preparation of rumen methane inhibitor |
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Application publication date: 20161207 |