CN116268190A - Application of seaweed or seaweed zymolyte in reducing methane emission of gastrointestinal tract - Google Patents
Application of seaweed or seaweed zymolyte in reducing methane emission of gastrointestinal tract Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/14—Pretreatment of feeding-stuffs with enzymes
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
- A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/10—Feeding-stuffs specially adapted for particular animals for ruminants
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- Animal Husbandry (AREA)
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- Biotechnology (AREA)
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Abstract
The invention discloses an application of seaweed or seaweed zymolyte as a feed additive in reducing methane production in the gastrointestinal tract of ruminants and in preparing feed with the effect of reducing methane production in the gastrointestinal tract of ruminants. The seaweed is selected from kelp, laver or enteromorpha. The seaweed zymolyte is prepared by the following method: adding seaweed into water, adding enzyme for enzymolysis, and then adding acid protease for enzymolysis to obtain seaweed zymolyte. The invention also discloses a feed with the effect of reducing methane production in the gastrointestinal tract of ruminants, which consists of basic feed and seaweed or seaweed zymolyte. The seaweed or seaweed zymolyte is added into basic feed, so that the seaweed feed has a good methane emission reduction effect on gastrointestinal tracts. The invention has the advantages of wide sources of raw materials, low cost and remarkable effect of reducing methane emission from gastrointestinal tracts, has important value and significance for the methane emission reduction technology of animal husbandry in China, and has great industrialized application potential.
Description
Technical Field
The invention relates to application of seaweed or seaweed zymolyte as a feed additive in reducing methane emission of gastrointestinal tracts, belonging to the technical field of methane emission reduction of ruminants.
Background
Methane is the second largest greenhouse gas next to carbon dioxide and its artificial emissions contribute about 20% of the greenhouse effect. Agricultural production is a major source of methane man-made emissions, and only cattle-based ruminant feeding processes contribute 16% of the world's man-made methane emissions. Methane emissions from animal husbandry come mainly from three aspects: firstly, in the feed stacking and fermenting process; secondly, a fecal treatment process; thirdly, methane produced by fermentation in the rumen of ruminants is discharged to the atmosphere through ruminant action. The first two sources can collect methane as clean energy by arranging a closed fermentation system, but methane discharged by ruminant behaviors is difficult to collect completely in the process of raising, so that methane emission reduction technology of the gastrointestinal tract of ruminants is an important subject of methane emission reduction in animal husbandry.
At present, three technical strategies exist for methane emission reduction of ruminant gastrointestinal tracts, namely, concentrate is added, daily ration nutrition is adjusted, and feed quality is improved; secondly, probiotic bacteria are artificially added to change the microbial community structure of the gastrointestinal tract; thirdly, methane inhibitor is specially added into the feed. The first strategy has higher cost and less than 50% methane emission reduction efficiency, and the fault-type change of the ration essence-coarse ratio can also cause a series of negative hazards such as acute or subacute acidosis of rumen, microbial flora disorder of digestive tract, hoof diseases, ketosis and the like. The second strategy technique is still immature and long-term effect is unknown. The specific addition of methane inhibitors to feeds is considered to be the most benefit-cost advantageous methane abatement strategy. Vegetable oils containing unsaturated fatty acids, vegetable extracts containing tannins and saponins, 3-nitrooxypropanol, nitrates and the like have been tried successively as ruminant feed additives, but their gastrointestinal methane emission reduction efficiency is rarely higher than 50%, and accumulation of these substances in ruminants causes food safety risks to raise concerns.
Disclosure of Invention
According to the invention, aiming at the prior art, experimental researches show that after seaweed or seaweed zymolyte after enzymolysis is added into feed, the methane content in the gastrointestinal tract can be effectively reduced after ruminant is fed, and a good methane emission reduction effect of the gastrointestinal tract is achieved.
The invention is realized by the following technical scheme:
the seaweed or seaweed zymolyte is used as a feed additive for reducing the production of methane in the gastrointestinal tract of ruminants, and is used for preparing feed with the effect of reducing the production of methane in the gastrointestinal tract of ruminants.
Further, the seaweed is one or more selected from kelp, laver and enteromorpha.
Further, the seaweed product is in the form of one or more of fresh, dried or freeze-dried powder.
Further, the seaweed zymolyte is prepared by the following method: pulverizing sea algae (fresh, dried or lyophilized powder), adding into water, and homogenizing; then adding enzyme, and carrying out enzymolysis under proper enzymolysis conditions to obtain a seaweed primary enzymolysis product; then adding acid proteinase, and performing enzymolysis under proper enzymolysis conditions to obtain seaweed zymolyte (which can be further sprayed to obtain dry powder). The enzyme is one or more than two of cellulase, pectase, neutral protease or amylase. Preferably, the enzyme is a complex enzyme, and consists of cellulase, pectase, neutral protease and amylase, wherein the enzyme activity ratio of the cellulase, pectase, neutral protease and amylase is 6:1.5:1:0.8.
The enzymes used in the invention are all commercial proteases known in the prior art, for example, the cellulase can be selected from cellulase R-10, the pectinase can be selected from pectinase Y-23, the neutral protease can be selected from microbial neutral protease, the amylase can be selected from Taka-amylase, and the acid protease can be selected from aspergillus protease I. Suitable enzymatic conditions for these commercial proteases (including enzyme addition, enzymatic hydrolysis temperature, time) can be found in the commercial specifications. In general, the enzymolysis temperature is controlled at 30-50 ℃ and the enzymolysis time is 6-72 hours.
Further, the addition amount of the seaweed or seaweed zymolyte in the feed is 0.2-10% of that of the basic feed, and the weight percentage is calculated by dry weight. Preferably 1% to 5%, more preferably 1% to 5%.
Further, the ruminant is selected from a bovine.
A feed for reducing methane production in ruminant gastrointestinal tract is prepared from basic feed and seaweed or seaweed zymolyte, wherein the addition amount of seaweed or seaweed zymolyte in feed is 0.2-10% by weight based on dry weight. Preferably 1% to 5%, more preferably 1% to 5%.
The basal feed is selected from the feeds suitable for ruminants, such as silage, which are known in the prior art.
The preparation method of the feed with the effect of reducing methane production in the gastrointestinal tract of ruminants comprises the following steps: adding the seaweed granule or seaweed zymolyte into basic feed, and mixing.
The seaweed particles can be prepared by the following method: pulverizing sea algae (fresh, dried or lyophilized powder).
According to the invention, seaweed or seaweed zymolyte is added into basic feed, and experimental researches show that the seaweed feed has a good methane emission reduction effect on gastrointestinal tracts. The seaweed used in the invention is kelp, laver or enteromorpha, wherein the kelp and the laver are rich seaweed cultivation resources in China, and the seaweed cultivation method has mature industrialized cultivation technology and is easy to obtain resources; enteromorpha prolifera is a main cause of green tide of yellow sea harmful algal bloom in China, and the method enriches the way of recycling the green tide. The invention has the advantages of wide sources of raw materials, low cost and remarkable effect of reducing methane emission from gastrointestinal tracts, has important value and significance for the methane emission reduction technology of animal husbandry in China, and has great industrialized application potential.
Detailed Description
The invention is further illustrated below with reference to examples. However, the scope of the present invention is not limited to the following examples. Those skilled in the art will appreciate that various changes and modifications can be made to the invention without departing from the spirit and scope thereof.
The instruments, reagents, materials, etc. used in the examples described below are conventional instruments, reagents, materials, etc. known in the art, and are commercially available. The experimental methods, detection methods, and the like in the examples described below are conventional experimental methods, detection methods, and the like that are known in the prior art unless otherwise specified.
Example 1 study of methane emission reduction in kelp and Porphyra
In the embodiment, the methane emission reduction effect of kelp and laver is detected by in vitro culture of rumen fluid of adult beef cattle, and the specific process is as follows:
gastric juice was obtained from rumen of 2-age cattle (body weight 517.5 kg).
Corn silage (obtained from livestock farm in Weihai city) is taken as basic feed, 5.0% of kelp or 1.0% of laver is added, dried kelp is commercial kelp (Phaeophyta kelp family, producing area: fujian), dried laver is commercial laver (red algae red makino rape family, producing area: fujian), and the dried kelp is crushed by a crusher without particle size requirement. A total of 3 experimental groups were set, each group being set with 3 parallel experiments: 5.0% of kelp group and 1.0% of laver group, and the basal feed (without seaweed added) is used as a control group.
On day 1 of the experiment, 2g of basic feed, 0.1g of kelp, 1.0% of laver and 0.02g of laver were added to 200ml of gastric juice, and sealed culture was performed. The experiment was continued for 1 month (30 days), and the total methane production of each experimental group was measured on days 7 and 30, and the methane content of the produced gas was measured by using a gas chromatograph (GC-FID, PEAK, USA) equipped with an FID detector, and the methane content in the produced gas was measured on-line.
Results: after 7 days, the methane yield of the control group was 899.0.+ -. 54.0mg, the methane yield of the 5.0% band group was 50.2.+ -. 19.4mg, the methane yield was 94.4% less, and the methane yield of the 1.0% band group was 51.0.+ -. 33.0mg, the methane yield was 94.3% less.
30, the methane yield of the control group was 794.1 + -117.3 mg, the methane yield of 5.0% of the kelp group was 388.4+ -153.4 mg, which was reduced by 51.1% (the consumption of the effective methane-reducing component of the kelp was depleted and the methane yield was gradually increased with the lapse of the culture time), and the methane yield of 1.0% of the laver group was 42.0+ -9.1 mg, which was reduced by 94.7%.
Example 2 methane emission reduction study of Enteromorpha prolifera
In this embodiment, the methane emission reduction effect of enteromorpha is detected by in vitro culture of rumen fluid of adult beef cattle, and the specific process is as follows:
gastric juice was obtained from rumen of 2-age cattle (body weight 517.5 kg).
Corn silage (obtained from livestock farm in Weihai city) is taken as basic feed, and 5.0% enteromorpha freeze-dried powder is added. A total of 2 experimental groups were set, each group being set with 3 parallel experiments: 5.0% Enteromorpha prolifera group, and basic feed (without seaweed) is used as control group.
The enteromorpha is collected in 36 degrees 23' N of Qingdao Bay in 7 and 10 days of 2022; the preparation method of the 120 DEG 47' E enteromorpha freeze-dried powder comprises the following steps:
(1) Soaking enteromorpha in clear water to reduce the salt content, fishing out, airing the enteromorpha in a shade until no water is dripped, and cutting the enteromorpha into 2-3 cm fragments by using scissors.
(2) And (3) performing vacuum freeze drying treatment on the enteromorpha sample by using a freeze dryer, wherein the drying pressure is 0Pa, and the drying temperature is-80 ℃.
(3) Processing the freeze-dried enteromorpha into enteromorpha freeze-dried powder by using a pulverizer, wherein the rotation speed of the pulverizer is 5000 rpm, and the pulverizing time is 20 minutes, so as to prepare the enteromorpha freeze-dried powder.
On the 1 st day of the experiment, 2g of basic feed and 0.1g of enteromorpha are added into 200ml of gastric juice, and the mixture is subjected to sealed culture. The experiment was continued for 1 month (30 days), and the total methane production of each experimental group was measured on days 7 and 30, and the average value was taken (methane content in the produced gas was measured using a gas chromatograph (GC-FID, PEAK, USA) equipped with an FID detector).
Results: after 7 days, the methane yield of the control group is 899.0+/-54.0 mg, and the methane yield of the 5.0% enteromorpha group is 489.0 +/-193.1 mg, which is reduced by 45.6%. After 30 days, the methane yield of the control group is 794.1 +/-117.3 mg, and the methane yield of the 5.0% enteromorpha group is 458.3+/-104.7 mg, which is reduced by 42.3%.
EXAMPLE 3 preparation of kelp enzymatic hydrolysate
The method comprises the following steps:
(1) Soaking dried herba Zosterae Marinae (dry herba Zosterae Marinae, commercial herba Zosterae Marinae, phaeophyta family, and Fujian) in clear water, removing sludge on herba Zosterae Marinae, and reducing its salt content; fishing out the kelp, crushing the kelp, and adding water with the mass of 4 times of that of the kelp to prepare the kelp homogenate. Adding complex enzyme into the kelp homogenate, wherein the adding amount of the complex enzyme is that 4g of complex enzyme is added into each 1L of kelp homogenate; the complex enzyme consists of cellulase R-10, pectase Y-23, microbial neutral protease and Taka-amylase, and the enzyme activity ratio of the cellulase R-10 to the pectase Y-23 to the microbial neutral protease to the Taka-amylase is 6:1.5:1:0.8. And (3) performing enzymolysis for 72 hours at 42 ℃ to obtain the primary kelp enzymolysis liquid.
(2) Filtering the primary enzymolysis liquid of the kelp by using a backflushing extrusion filter at the filtering temperature of 10 ℃, obtaining clear kelp liquid after filtering, concentrating the clear kelp liquid at the temperature of 55 ℃ to obtain concentrated clear liquid, wherein the content of water insoluble matters in the concentrated clear liquid is 5g/L.
(3) Adding acid protease (Aspergillus protease I) (enzyme activity 100000 U.g) -1 ) The addition amount of the acid protease was 1% by weight of the concentrated supernatant. The pH value of the concentrated clear liquid is regulated to 6.7 by hydrochloric acid, and the concentrated clear liquid is subjected to enzymolysis for 12 hours at 55 ℃ to obtain a kelp compound enzymolysis liquid which is rich in laminarin, kelp monosaccharide, kelp oligosaccharide and mannitol, small molecular organic acid mainly containing laminaric acid and active compound enzyme used for enzymolysis compared with a kelp dry product.
Meanwhile, the invention also prepares another 5 kelp enzymolysis solutions, and the preparation method is the same as the above, and the difference is that: replacing the complex enzyme in the step (1) with cellulase, pectase, protease and amylase and complex enzyme 2 (comprising cellulase, pectase, protease and amylase, wherein the enzyme activity ratio of the cellulase, pectase, protease and amylase is 1:1:1). The obtained kelp enzymatic hydrolysate is respectively called cellulase enzymatic hydrolysate, pectase enzymatic hydrolysate, protease enzymatic hydrolysate, amylase enzymatic hydrolysate and complex enzyme 2 enzymatic hydrolysate.
EXAMPLE 4 methane emission reduction study of kelp enzymatic hydrolysate
In the embodiment, the methane emission reduction effect of kelp zymolyte is detected by in-vitro culture of adult beef cattle rumen fluid, and the specific process is as follows:
gastric juice was obtained from rumen of 2-age cattle (body weight 517.5 kg).
Experiment 1: corn silage (obtained from livestock farm in wehai) is used as basic feed, 1.0%, 5.0% and 10% kelp complex enzymatic hydrolysate (based on dry weight) are added respectively, and 4 experimental groups (3 parallel experiments are arranged in each experimental group) are arranged in total: 1.0% of kelp compound enzymatic hydrolysate group, 5.0% of kelp compound enzymatic hydrolysate group and 10% of kelp compound enzymatic hydrolysate group, and taking basic feed (without kelp compound enzymatic hydrolysate) as a control group.
On the 1 st day of the experiment, 2g of basic feed, 1.0% of kelp compound enzymolysis liquid group, 5.0% of kelp compound enzymolysis liquid group and 10% of kelp compound enzymolysis liquid group are added into 200ml of gastric juice, and 0.02g, 0.1g and 0.2g of kelp compound enzymolysis liquid are respectively added, and the mixture is sealed and cultivated. The experiment was continued for 1 month (30 days), and the total methane production of each experimental group was measured on days 7 and 30 and averaged.
Results: after 7 days, the methane yield of the control group was 899.0.+ -. 54.0mg. The methane yield of the 1.0 percent kelp compound enzymolysis liquid group, the 5.0 percent kelp compound enzymolysis liquid group and the 10 percent kelp compound enzymolysis liquid group is 78.1 plus or minus 23.7mg, 44.7 plus or minus 11.3mg and 95.8 plus or minus 50.4mg respectively, and the methane emission reduction efficiency is 91.3 percent, 95.0 percent and 89.3 percent respectively.
After 30 days, the methane yield of the control group was 794.1.+ -. 117.3mg. The methane yield of the 1.0 percent kelp compound enzymolysis liquid group, the 5.0 percent kelp compound enzymolysis liquid group and the 10 percent kelp compound enzymolysis liquid group are respectively 31.3+/-10.7 mg, 5.7+/-2.5 mg and 38.1+/-15.7 mg, and the methane emission reduction efficiency is respectively 96.1 percent, 99.3 percent and 95.1 percent.
Experiment 2: taking corn silage as basic feed, adding 5.0% kelp enzymatic hydrolysate (based on dry weight) for enzymatic hydrolysis by different enzymes, and setting 7 experimental groups (3 parallel experiments are set for each experimental group): 5.0% of kelp complex enzyme hydrolysate group, 5.0% of cellulase (cellulase R-10) enzyme hydrolysate group, 5.0% of pectase (pectase Y-23) enzyme hydrolysate group, 5.0% of protease (microorganism neutral protease) enzyme hydrolysate group, 5.0% of amylase (Taka-amylase) enzyme hydrolysate group and 5.0% of complex enzyme 2 (consisting of cellulase R-10, pectase Y-23, microorganism neutral protease and Taka-amylase, and the enzyme activity ratio of cellulase R-10, pectase Y-23, microorganism neutral protease and Taka-amylase is 1:1:1:1) enzyme hydrolysate group, and the basic feed (without adding the kelp complex enzyme hydrolysate) is taken as a control group.
On the 1 st day of the experiment, 2g of basic feed, 5.0% of kelp complex enzymatic hydrolysate group, 5.0% of cellulase enzymatic hydrolysate group, 5.0% of pectase enzymatic hydrolysate group, 5.0% of protease enzymatic hydrolysate group, 5.0% of amylase enzymatic hydrolysate group and 5.0% of complex enzyme 2 enzymatic hydrolysate group are added into 200ml of gastric juice, and 0.1g of each kelp enzymatic hydrolysate is added respectively for sealing culture. The experiment was continued for 1 month (30 days), and the total methane production of each experimental group was measured on days 7 and 30 and averaged.
Results: after 7 days, the methane yield of the control group was 899.0.+ -. 54.0mg. The methane yield of the 5.0% kelp compound enzymolysis liquid group is 44.7+/-11.3 mg, and the methane emission reduction efficiency is 95.0%. The yields of the 5.0% cellulase enzymolysis liquid group, the 5.0% pectase enzymolysis liquid group, the 5.0% protease enzymolysis liquid group, the 5.0% amylase enzymolysis liquid group and the 5.0% compound enzyme 2 enzymolysis liquid group are respectively 46.8+/-17.4 mg, 48.5+/-20.6 mg, 50.3+/-15.4 mg, 50.3+/-12.9 mg and 45.0+/-8.7 mg, and the emission reduction efficiencies are respectively 94.8%, 94.6%, 94.4% and 95.0%.
After 30 days, the methane yield of the control group was 794.1.+ -. 117.3mg. The methane yield of the 5.0% kelp compound enzymolysis liquid group is 5.7+/-2.5 mg, and the methane emission reduction efficiency is 99.3%. The yields of the 5.0% cellulase enzymolysis liquid group, the 5.0% pectase enzymolysis liquid group, the 5.0% protease enzymolysis liquid group, the 5.0% amylase enzymolysis liquid group and the 5.0% compound enzyme 2 enzymolysis liquid group are respectively 176.3+/-37.0 mg, 235.8+/-43.8 mg, 288.3 +/-57.1 mg, 302.6+/-55.3 mg and 99.3+/-23.9 mg, and the methane emission reduction efficiencies are respectively 77.8%, 70.3%, 63.7%, 61.9% and 87.5%.
Conclusion: after enzymolysis, compared with the dried kelp product, the methane emission reduction effect is more obvious. As can be seen from the comparison of the effects of the enzymatic hydrolysate of different enzymes, the specific complex enzyme (comprising cellulase R-10, pectinase Y-23, microbial neutral protease and Taka-amylase, the enzyme activity ratio of the cellulase R-10, the pectinase Y-23, the microbial neutral protease and the Taka-amylase is 6:1.5:1:0.8) has the best effect (the effect after 7 days is almost the same, but the effect after 30 days is obviously different), and the specific complex enzyme (comprising cellulase R-10, pectinase Y-23, microbial neutral protease and Taka-amylase, the enzyme activity ratio of the cellulase R-10, the pectinase Y-23, the microbial neutral protease and the Taka-amylase is 1:1:1) is obviously better than that of the single enzyme.
The foregoing examples are provided to fully disclose and describe how to make and use the claimed embodiments by those skilled in the art, and are not intended to limit the scope of the disclosure herein. Modifications that are obvious to a person skilled in the art will be within the scope of the appended claims.
Claims (10)
1. The use of seaweed or seaweed hydrolysate as feed additive for reducing methane production in the gastrointestinal tract of ruminants or for preparing a feed having the effect of reducing methane production in the gastrointestinal tract of ruminants.
2. The use according to claim 1, characterized in that: the seaweed is selected from one or more of kelp, laver and enteromorpha.
3. The use according to claim 1, characterized in that: the seaweed product is in the form of one or more of fresh product, dried product or lyophilized powder.
4. Use according to claim 1 or 2 or 3, wherein the seaweed hydrolysate is prepared by: pulverizing sea algae, adding into water, and homogenizing; then adding enzyme for enzymolysis to obtain seaweed primary zymolyte; then adding acid protease for enzymolysis to obtain seaweed zymolyte; the enzyme is one or more than two of cellulase, pectase, neutral protease or amylase.
5. The use according to claim 4, characterized in that: the enzyme is a complex enzyme and consists of cellulase, pectase, neutral protease and amylase, wherein the enzyme activity ratio of the cellulase, pectase, protease and amylase is 6:1.5:1:0.8.
6. The use according to claim 1, characterized in that: the addition amount of the seaweed or seaweed zymolyte in the feed is 0.2-10 percent, and the seaweed or seaweed zymolyte accounts for the weight percentage based on dry weight.
7. A feed having the effect of reducing methane production in the gastrointestinal tract of ruminants, characterized by: the feed consists of basic feed and seaweed or seaweed zymolyte, wherein the addition amount of the seaweed or seaweed zymolyte in the feed is 0.2-10 percent, and the weight percentage is calculated by dry weight.
8. The feed with the effect of reducing methane production in the gastrointestinal tract of ruminants according to claim 7, wherein: the basal feed is selected from corn silage.
9. A method for preparing a feed having the effect of reducing methane production in the gastrointestinal tract of ruminants in accordance with claim 7, wherein: adding the seaweed granule or seaweed zymolyte into basic feed, and mixing.
10. A method for preparing a feed having the effect of reducing methane production in the gastrointestinal tract of ruminants in accordance with claim 7, wherein:
the seaweed particles are prepared by the following method: pulverizing Sargassum to obtain the final product;
the seaweed zymolyte is prepared by the following method: pulverizing sea algae, adding into water, and homogenizing; then adding enzyme for enzymolysis to obtain seaweed primary zymolyte; then adding acid protease for enzymolysis to obtain seaweed zymolyte; the enzyme is one or more than two of cellulase, pectase, neutral protease or amylase.
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