CN121369578A - Compound acidulant for improving feed intake of lactating sows and preparation method thereof - Google Patents
Compound acidulant for improving feed intake of lactating sows and preparation method thereofInfo
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- CN121369578A CN121369578A CN202511980239.4A CN202511980239A CN121369578A CN 121369578 A CN121369578 A CN 121369578A CN 202511980239 A CN202511980239 A CN 202511980239A CN 121369578 A CN121369578 A CN 121369578A
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Abstract
本发明涉及一种提高哺乳母猪采食量的复合酸化剂及其制备方法，属于饲料添加剂技术领域。该复合酸化剂包括壁材和芯材，所述壁材包括胱氨酸‑γ‑聚谷氨酸和精氨酸‑壳聚糖；所述芯材包括有机酸、有机酸盐和发酵液。本发明的复合酸化剂利用胱氨酸交联的γ‑聚谷氨酸和精氨酸接枝的壳聚糖作为壁材，通过包裹有机酸形成具有肠道靶向释放功能的微胶囊，能有效提高酸化剂的利用效率及哺乳母猪的采食量。
This invention relates to a compound acidifier for improving feed intake in lactating sows and its preparation method, belonging to the field of feed additive technology. The compound acidifier comprises a wall material and a core material. The wall material comprises cystine-γ-polyglutamic acid and arginine-chitosan; the core material comprises organic acids, organic acid salts, and fermentation broth. The compound acidifier of this invention utilizes cystine-crosslinked γ-polyglutamic acid and arginine-grafted chitosan as the wall material, forming microcapsules with intestinal-targeted release function by encapsulating organic acids, which can effectively improve the utilization efficiency of the acidifier and the feed intake of lactating sows.
Description
Technical Field
The invention belongs to the technical field of feed additives, and particularly relates to a compound acidifier for improving the feed intake of lactating sows and a preparation method thereof.
Background
The feed intake of lactating sows directly determines the lactation performance, the body condition recovery and the survival rate of piglets, and is a core factor influencing the economic benefit of a pig farm. In actual breeding, sows generally face the dilemma of insufficient feed intake after delivery, and further chain problems such as weight loss, lactation reduction and the like are caused. In order to solve the problem, the feed acidifier is widely used as a safe and green additive, and can inhibit harmful bacteria and promote digestion by reducing the pH value of the gastrointestinal tract, thereby indirectly improving the feed intake. The compound acidifier in the market at present is mostly prepared by adopting a simple physical mixing process of organic acid and carrier, but has a certain effect, is limited by the traditional technical thought, cannot break through the rough mode of 'mixing-releasing', has obvious short plates in the aspects of accuracy, stability and functionality, and is difficult to meet the requirements of modern high-efficiency cultivation.
The existing compound acidifier mainly has the key problems that firstly, targeting is extremely poor, organic acid is released prematurely in a stomach strong acid environment, active ingredients are consumed before reaching an intestinal tract, so that the antibacterial concentration at the rear end of the intestinal tract is insufficient, accurate regulation and control cannot be realized, secondly, stability is insufficient, small-molecule organic acid is easy to volatilize and high in corrosiveness, loss rate in a high-temperature and high-pressure process of feed processing and granulating is high, efficiency attenuation is obvious in long-term storage, corrosion is caused on processing equipment, thirdly, the compound acidifier has a single function, inhibits pathogenic bacteria only by relying on an acidic environment, lacks multiple synergistic mechanisms for promoting beneficial bacteria proliferation, repairing intestinal barrier and the like, and has limited effect in complex intestinal microecology. These defects severely restrict the practical effect of acidulants and become a technical pain point which is not solved for a long time in the industry.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a compound acidifier for improving the feed intake of lactating sows and a preparation method thereof, and the core innovation is to construct a microcapsule system taking cystine-crosslinked gamma-polyglutamic acid and arginine grafted chitosan as wall materials, and break through is realized by a triple response mechanism, wherein pH response is utilized to ensure that the arginine chitosan is not released in the stomach and swells in the intestinal tract, and the arginine chitosan is accurately released in a specific environment of the intestinal tract by means of the reduction response of disulfide bonds, and meanwhile, the mucous membrane adhesion and antibacterial synergistic effect of the arginine chitosan is exerted.
The technical scheme for realizing the purpose of the invention is as follows:
A compound acidifier for improving the feed intake of lactating sows comprises a wall material and a core material, wherein the wall material comprises cystine-gamma-polyglutamic acid and arginine-chitosan, the mass ratio of the cystine-gamma-polyglutamic acid to the arginine-chitosan in the wall material is 1 (0.8-1.2), and the core material comprises organic acid, organic acid salt and fermentation liquor.
The fermentation liquid comprises a composite microbial inoculum and a fermentation substrate, wherein the composite microbial inoculum comprises bacillus natto and saccharomycetes, the saccharomycetes comprise one or two of brewer's saccharomycetes and candida utilis, and the fermentation substrate comprises, by weight, 20-30 parts of soybean meal, 1-5 parts of bran, 0.5-2 parts of magnesium sulfate, 2-5 parts of dipotassium hydrogen phosphate, 2-5 parts of yeast extract powder and 60-70 parts of water.
The preparation method of the fermentation broth comprises the following steps:
Mixing the bacillus natto liquid and the saccharomycete liquid to prepare a compound microbial agent, inoculating the compound microbial agent into the fermentation substrate for aerobic fermentation, heating the fermentation liquid to terminate all microorganisms and enzyme activities after fermentation, centrifuging to obtain supernatant, and performing aseptic filtration to obtain the fermentation liquid.
The ratio of the bacillus natto liquid to the saccharomycete liquid is (1-3) 1, and the weight ratio of the composite microbial inoculum to the fermentation substrate is (1-8) 100.
A preparation method of a compound acidifier for improving the feed intake of lactating sows comprises the following steps:
S1, preparing a composite wall material solution and a core material solution, namely respectively dissolving cystine-gamma-polyglutamic acid and arginine-chitosan to prepare solutions, mixing the two solutions to obtain the composite wall material solution, weighing organic acid, organic acid salt and fermentation liquor, mixing the fermentation liquor with the organic acid salt firstly, and then slowly and uniformly mixing the fermentation liquor with the organic acid salt to obtain the core material solution;
S2, complex-phase emulsification and solidification, namely dripping core material solution into corn oil containing a surfactant, homogenizing at high speed under ice bath condition to form primary emulsion, dripping into composite wall material solution, continuously emulsifying, slowly dripping cross-linking agent into the composite emulsion, and stirring;
S3, purifying and collecting, namely centrifuging and washing the mixture obtained in the step S2, dispersing the obtained solid in a freeze-drying protective agent, and obtaining the compound acidifier after freeze-drying and sieving.
The mass ratio of the organic acid to the organic acid salt to the fermentation liquor is (2-4) 1:1.
The cross-linking agent is one or two selected from sodium tripolyphosphate and sodium citrate, and the freeze-drying protective agent is one or more selected from trehalose, mannitol and maltodextrin.
The invention also provides an application of the compound acidulant for improving the feed intake of the lactating sow, which is prepared by the preparation method of the compound acidulant for improving the feed intake of the lactating sow.
A method for improving the feed intake of lactating sows comprises the following steps of uniformly mixing a compound acidifier with a basic feed to obtain a compound acidifier feed for improving the feed intake of lactating sows, and feeding the compound acidifier feed to lactating sows.
The mass ratio of the compound acidulant to the basic diet is (0.5-2) 100.
Advantageous effects
The invention provides a composite acidulant for improving feed intake of lactating sows based on hydrogel microcapsules and a preparation method thereof. The compound acidifier disclosed by the invention uses the gamma-polyglutamic acid crosslinked by cystine and the arginine grafted chitosan as wall materials, and forms the microcapsule with the intestinal targeted release function by wrapping organic acid, so that the utilization efficiency of the acidifier and the feed intake of lactating sows can be effectively improved, and the compound acidifier comprises the following beneficial effects:
1. The microcapsule wall material prepared by the method has the triple intelligent release characteristics of pH response, reduction response and enzyme response, has a stable microcapsule structure in a stomach acidic environment, and has the advantages that disulfide bonds are broken, the wall material erodes and degrades when the concentration of glutathione is increased in a rear neutral environment in the intestinal tract, the sudden release effect of organic acid is realized, the acidification efficiency is accurately delivered to a main harmful bacteria enrichment region of the intestinal tract, and the ineffective consumption in the stomach is avoided.
2. The high-efficiency and slow-release compound acidifier has the advantages that the high-efficiency compound acidifier has high organic acid encapsulation efficiency based on a compact polyelectrolyte compound network formed by cystine-gamma-polyglutamic acid and arginine-chitosan, can effectively delay the diffusion rate of organic acid in storage and gastric juice, realizes programmed release through controllable degradation in intestinal tracts, enables the acidifier with unit dose to exert maximum efficiency, and is expected to be improved compared with the utilization rate of common acidifiers.
3. The scheme not only provides slow release acidification function of organic acid, but also generates synergistic effect with broad-spectrum antibacterial activity of arginine-chitosan, and multiplies inhibition effect on pathogenic bacteria such as escherichia coli and salmonella, and meanwhile, gamma-polyglutamic acid and other components can be used as prebiotics to promote proliferation of beneficial bacteria such as lactobacillus, optimize intestinal microecological balance and lighten intestinal immune load.
4. On one hand, the arginine modified chitosan has excellent mucous membrane adhesion, can prolong the acting time and promote the absorption of nutrient substances, and on the other hand, the healthy intestinal tract environment can promote the digestion and absorption rate of the nutrient, thereby stimulating the sow to eat the desire to meet the requirements of lactation and body condition recovery.
5. The wall material and the modifier are natural sources, have good biocompatibility and no toxic or side effect, and the in-vivo metabolites are water and carbon dioxide, so that the wall material and the modifier have no residual risk and meet the requirements of green healthy cultivation.
6. The fermented flavor substances are introduced, so that the palatability and the feeding attraction of the sow to the compound feed can be improved, the digestion and the absorption of the sow to the feed can be further improved, the feed intake can be improved, the lactation can be promoted, and the survival rate of piglets can be improved.
In conclusion, the intelligent targeting hydrogel microcapsule is constructed by the natural composite wall material, the advantages of multiple cores are integrated, the postpartum intestinal health and the feeding pain point of the lactating sow are accurately solved, the acidulant efficiency and the nutrient absorption can be improved, the intestinal microecology can be optimized, the cultivation safety is guaranteed, and the intelligent targeting hydrogel microcapsule is suitable for large-scale production.
Drawings
FIG. 1 is an infrared spectrum of chitosan and arginine-chitosan.
FIG. 2 is an infrared spectrum of gamma-polyglutamic acid and cystine-gamma-polyglutamic acid.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the examples, the experimental methods used are conventional methods unless otherwise specified, and the materials, reagents, etc. used, unless otherwise specified, are commercially available.
The raw materials and equipment used in the examples and comparative examples are now described as follows, eq representing molar equivalents:
Chitosan has a molecular weight of 50 kDa and a degree of deacetylation of 85% and is available from Shanghai Yi En chemical technology Co.
The molecular weight of the gamma-polyglutamic acid is 50000-100000, and the gamma-polyglutamic acid is purchased from Shanghai Yi En chemical technology Co.
Yeast, beer yeast, commercially available.
Arginine-chitosan self-made by the following preparation method:
Preparing 0.1 mol/L MES buffer solution with pH of 6 as a solvent, adding 0.3 eq arginine, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride with the molar weight of 1.2 times that of the arginine and N-hydroxysuccinimide with the molar weight of 1.2 times that of the arginine under stirring, activating carboxyl groups for half an hour in the dark, dissolving 1.0 eq chitosan with the molar weight of amino groups in 1% acetic acid aqueous solution in another container to prepare 1% uniform solution, slowly dripping the activated arginine solution into the chitosan solution under stirring, regulating and maintaining the pH=5.5 with 0.1 mol/L sodium hydroxide solution, stirring for reaction 6 h at room temperature in the dark, adding stirring 15min after the reaction is finished, dialyzing the reaction solution against deionized water with a dialysis bag with the molecular weight of 10 kDa, changing deionized water every 12 h, and performing vacuum freeze drying to obtain arginine-chitosan.
The preparation method of the cystine-gamma-polyglutamic acid comprises the following steps:
0.1 eq cystine was dissolved in 0.1 mol/L HEPES buffer pH 8, adding 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride in an amount which is 1.2 times the molar amount relative to cystine and N-hydroxysuccinimide in an amount which is 1.2 times the molar amount relative to cystine, and reacting at 25 ℃ for 30 min; subsequently, the first and second heat exchangers are connected, 1.1 eq glycine ethyl ester hydrochloride is added into the reaction system, the pH value is regulated to 8 by 0.1 mol/L sodium hydroxide solution, the reaction is continued for 30 min, converting activated carboxyl into a 'cystine-glycine ethyl ester' dipeptide structure through aminolysis, transferring the reaction solution into a dialysis bag with a molecular weight cut-off of 1 kDa, dialyzing 6 h with deionized water, removing small molecular byproducts to obtain purified blocked cystine solution, realizing carboxyl blocking, while ensuring that the amino group of cystine remains free; in a separate vessel, 1.0 eq. Gamma. -polyglutamic acid, calculated as the number of carboxyl groups moles, was dissolved in 0.1 mol/L MES buffer pH 6.0, activated with 1.0 eq 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 1.0 eq N-hydroxysuccinimide at 25℃for 30 min; finally, the step of obtaining the product, dripping the blocked cystine solution into an activated gamma-polyglutamic acid solution, cooling the activated gamma-polyglutamic acid solution to 0 ℃ in an ice bath, slowly dripping the blocked cystine solution into the mixture under intense stirring, wherein the dripping time is longer than 1h, and after the dripping is finished, the reaction is continued at 0 ℃ for 2h, then the reaction is transferred to room temperature for 4h, glycine is added for quenching reaction, and the cystine-gamma-polyglutamic acid is obtained after dialysis purification and freeze drying of the reaction liquid.
Fermentation broth 1: the preparation method comprises the following steps:
Weighing 25 parts of soybean meal, 3 parts of bran, 1.5 parts of magnesium sulfate, 3 parts of dipotassium hydrogen phosphate and 3.5 parts of yeast extract powder according to the proportion, uniformly mixing the materials with 65 parts of water, regulating the pH of the mixture to 7.5 by using a 1 mol/L sodium hydroxide solution, sterilizing the mixture at high pressure of 121 ℃ to 25. min, cooling the mixture to below 35 ℃ for standby, mixing the revived and expanded bacillus natto liquid with the activated saccharomycetes liquid according to the ratio of the viable bacteria number of 2:1, preparing a composite microbial agent with the total viable bacteria number of more than or equal to 1.0x10 8 CFU/mL, inoculating the composite microbial agent into the sterilized fermentation substrate with 5% of the total mass of the fermentation substrate under aseptic operation, uniformly stirring the mixture, performing aerobic fermentation at 35 ℃ for 50 h, performing 200 rpm stirring and 0.8 vvm to maintain dissolved oxygen, heating the fermentation liquid to 80 ℃ when the pH value is maintained stable by at least 2h, maintaining the fermentation liquid at 25 min to terminate all microorganisms and enzymes, and performing aseptic aeration on the fermentation liquid at 8000 x 15.45 mu.45 m, and filtering the fermentation liquid under the condition of being subjected to aseptic aeration of 62 m to filtering to obtain a filter membrane.
The fermentation liquor 2 is prepared by self-making, and the preparation method is different from the fermentation liquor 1 in that the proportion of fermentation substrates is modified into 20 parts of soybean meal, 1 part of bran, 0.5 part of magnesium sulfate, 2 parts of dipotassium hydrogen phosphate, 2 parts of yeast extract powder and 60 parts of water, and other conditions are unchanged, so that the fermentation liquor 2 is obtained.
The fermentation liquor 3 is self-made, and the preparation method is different from the fermentation liquor 1 in that the proportion of fermentation substrates is modified to 30 parts of soybean meal, 5 parts of bran, 2 parts of magnesium sulfate, 5 parts of dipotassium hydrogen phosphate, 5 parts of yeast extract powder and 70 parts of water, and other conditions are unchanged, so that the fermentation liquor 3 is obtained.
The simulated artificial gastric juice is self-made and the preparation method is as follows:
Taking 0.1 mol/L hydrochloric acid solution 16.4 mL, adding distilled water 800 mL and pepsin 10 g, and adding water to constant volume to 1000 mL for later use.
Simulating artificial intestinal juice: the preparation method comprises the following steps:
Adding 500 mL g of potassium dihydrogen phosphate into 500 mL g of distilled water, stirring to dissolve completely, adjusting pH to 6.8 with 0.4% sodium hydroxide solution, sterilizing at 121deg.C to 20. 20 min, collecting pancreatic juice 10 g, adding sterile water to dissolve, mixing the two solutions, and diluting with sterile water to 1000 mL.
Basic diet, namely corn-soybean meal type diet prepared according to NRC (1998) pig feeding standard, wherein the composition and the nutrition level of the basic diet are shown in table 1:
TABLE 1 basal diet composition and nutrient level (air-dried basis)
Wherein, the premix provides vitamin A2000 IU, vitamin D200 IU, vitamin E45 IU, vitamin K0.5 mg, vitamin B1 1 mg, vitamin B6.85 mg, vitamin B12 15 mg, pantothenic acid 12 mg, nicotinic acid 10.25 mg, folic acid 1.35 mg, biotin 0.21 mg, manganese 20 mg, iron 100 mg, copper 20 mg, iodine 0.14 mg and selenium 0.15 mg for each kilogram of feed.
Examples
Example 1
The composite acidulant 1 is prepared by self-making and the preparation method is as follows:
S1, preparing a composite wall material solution 1 and a core material solution 1, namely weighing 5g cystine-gamma-polyglutamic acid, preparing a 1% uniform solution by using a PBS buffer solution with the pH of 0.1 mol/L and 6.5, stirring to fully dissolve the solution, weighing 5g arginine-chitosan, dissolving the solution by using a 1% acetic acid aqueous solution, preparing a 1% uniform solution, stirring to fully dissolve the solution, regulating the pH of the solution to 5.5 by using a 0.1 mol/L sodium hydroxide solution, mixing the two solutions according to the volume ratio of 1:1 to obtain a composite wall material solution 1 with the total polymer concentration of 1%, regulating the pH of 5.5 for standby, weighing formic acid, ammonium formate and a fermentation solution 1 according to the mass ratio of 3:1:1, mixing the fermentation solution and the ammonium formate in an ice bath and a fume hood, slowly adding the formic acid, stirring and uniformly mixing the mixture to obtain the core material solution 1;
S2, complex phase emulsification and solidification, namely slowly dripping the core material solution 1 into corn oil containing 3% Span-80, homogenizing at a high speed by 13000 rpm under ice bath condition to form colostrum, slowly dripping the colostrum into the composite wall material solution 1 at a stirring speed of 800 rpm, after dripping, raising the stirring speed to 1500 rpm, continuously emulsifying 10min, slowly dripping an equal volume of 0.5% sodium tripolyphosphate aqueous solution into the composite emulsion under 800 rpm stirring, wherein the dripping time is not less than 20 min, and continuously stirring and crosslinking 2 h;
S3, sub-packaging the reaction solution, centrifuging 1500 rpm for 3 minutes in a fume hood, discarding the supernatant, washing 2 times with petroleum ether to remove residual corn oil, washing once with absolute ethyl alcohol, washing 2 times with deionized water, centrifuging, repeating 2 times, washing 2 times with deionized water, re-dispersing the washed microspheres in a 5% aqueous solution of trehalose with the concentration being 2 times of the microsphere volume, sub-packaging the suspension in a freeze-drying bottle, pre-freezing at-80 ℃ for 6 h, then drying 24 h in a freeze dryer under the conditions of-50 ℃ and vacuum degree of less than 10 Pa until the microspheres are completely dried, passing the dried powder through a 200-target standard sieve, collecting the undersize to obtain the compound acidifier 1, and preserving in a dark place, sealing and moisture-proof manner.
Example 2
The composite acidulant 2 is prepared by self-making, and the preparation method is compared with the composite acidulant 1, wherein the difference is that the fermentation liquor 1 in the step S1 is replaced by the fermentation liquor 2, and other conditions are unchanged, so that the composite acidulant 2 is obtained.
Example 3
The composite acidulant 3 is prepared by self-making, and the preparation method is compared with the composite acidulant 1, wherein the difference is that the fermentation liquor 1 in the step S1 is replaced by the fermentation liquor 3, and other conditions are unchanged, so that the composite acidulant 3 is obtained.
Example 4
The composite acidulant 4 is prepared by self-making, and the preparation method is compared with the composite acidulant 1, wherein the difference is that the mass ratio of formic acid to ammonium formate to the fermentation liquor 1 in the step S1 is replaced by 2:1:1, and other conditions are unchanged, so that the composite acidulant 4 is obtained.
Example 5
The composite acidulant 5 is prepared by self-making, and the preparation method is compared with the composite acidulant 1, wherein the difference is that the mass ratio of formic acid to ammonium formate to fermentation liquor 1 in the step S1 is replaced by 4:1:1, and other conditions are unchanged, so that the composite acidulant 5 is obtained.
Comparative example 1
The composite acidulant 6 is prepared by self-making, and the preparation method is different from the composite acidulant 1 in that cystine-gamma-polyglutamic acid is replaced by gamma-polyglutamic acid, and other conditions are unchanged, so that the composite acidulant 6 is obtained.
Comparative example 2
The composite acidulant 7 is prepared by self-making, and the preparation method is different from the composite acidulant 1 in that arginine-chitosan is replaced by chitosan and other conditions are unchanged, so that the composite acidulant 7 is obtained.
Comparative example 3
The compound acidulant 8 is commercially available from Weifang midday feed technology Co.
Application example
Application example 1
The composite acidulant diet 1 is self-made and is prepared by mixing the composite acidulant 1 with basic diet according to the weight ratio of 1:100 to serve as the composite acidulant diet 1.
Application example 2
The composite acidifier feed 2 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is mixed with the basic feed according to the weight ratio of 1:200, and other conditions are unchanged, so that the composite acidifier feed 2 is obtained.
Application example 3
The composite acidifier feed 3 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is mixed with the basic feed according to the weight ratio of 1:50, and other conditions are unchanged, so that the composite acidifier feed 3 is obtained.
Application example 4
The composite acidifier feed 4 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 2, and other conditions are unchanged, so that the composite acidifier feed 4 is obtained.
Application example 5
The composite acidulant feed 5 is self-made, and the preparation method is different from the composite acidulant feed 1 in that the composite acidulant 1 is replaced by the composite acidulant 3, and other conditions are unchanged, so that the composite acidulant feed 5 is obtained.
Application example 6
The composite acidifier feed 6 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 4, and other conditions are unchanged, so that the composite acidifier feed 6 is obtained.
Application example 7
The composite acidifier feed 7 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 5, and other conditions are unchanged, so that the composite acidifier feed 7 is obtained.
Comparative application example 1
The composite acidifier feed 8 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 6, and other conditions are unchanged, so that the composite acidifier feed 8 is obtained.
Comparative application example 2
The composite acidifier feed 9 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 7, and other conditions are unchanged, so that the composite acidifier feed 9 is obtained.
Comparative application example 3
The composite acidifier feed 10 is self-made, and the preparation method is different from the composite acidifier feed 1 in that the composite acidifier 1 is replaced by the composite acidifier 8, and other conditions are unchanged, so that the composite acidifier feed 10 is obtained.
The following is a test method related to performance parameters in the present invention:
1. Fourier transform infrared spectrum (FT-IR) is tested by adopting a Thermo Nicolet IS model Fourier transform infrared spectrometer, wherein FIG. 1 shows the infrared spectrograms of chitosan and arginine-chitosan, the arginine chitosan has an amide I band characteristic peak and an amide II band characteristic peak at 1650 cm -1 and 1590 cm -1, the newly generated absorption peak covers the amino characteristic peak of the original chitosan, which indicates successful preparation of the arginine-chitosan, and FIG. 2 shows the infrared spectrograms of gamma-polyglutamic acid and cystine-gamma-polyglutamic acid, the remarkable enhancement of the amide I band characteristic peak and the amide II band characteristic peak indicates successful synthesis of-CONH-crosslinking bond, which proves successful synthesis of cystine-gamma-polyglutamic acid.
2. Testing the stability of the compound acidifier, namely testing the compound acidifiers of examples 1-5 and comparative examples 1-2 by using simulated artificial gastric fluid and simulated artificial intestinal fluid according to a second method (a paddle method) of the four-part rule 0931 of Chinese pharmacopoeia 2020, placing the compound acidifiers of examples 1-5 and comparative examples 1-2 into a dissolution cup containing 200 mL and 37 ℃ constant temperature artificial gastric fluid, continuously oscillating for 2h at the rotating speed of 100 rpm, immediately taking 1 mL solution, filtering the solution by using a 0.22 mu m organic filter membrane, removing hydrochloric acid from the filtrate by nitrogen blowing, re-dissolving the filtrate by using methanol, measuring the content of the representative core material formic acid by using a high performance liquid chromatography, and calculating the gastric fluid retention rate according to the formula retention rate (%) = (Ct/C0) x 100, wherein Ct is the measurement concentration after the gastric fluid is treated by 2h, and C0 is the original content of formic acid; subsequently, the whole gastric juice-treated compound acidifier was transferred to 200 mL and 37 ℃ constant-temperature artificial intestinal juice, and the mixture was continuously oscillated at 100 rpm, and was automatically sampled at time points of 5, 15, 30, 60 and 120min for 2mL, while the isothermal equivalent volume of fresh artificial intestinal juice was supplemented, and the sample was filtered through a 0.22 μm filter membrane, and then directly subjected to hplc analysis for the content of formic acid, and the cumulative release of intestinal juice was calculated according to the formula cumulative release (%) = (cumulative release/total dose) ×100, and the results are shown in table 2 below.
3. The test of the growth performance index of the lactating sows is that the test is developed in a pig farm, 50 healthy sows with 3-5 batches and approximate expected delivery period are randomly divided into 10 groups, each group of sows is transferred into a delivery house three days before the expected delivery period, the delivery house is a high-bed and slotted floor, the ventilation of the house is good, the interior of the house is kept clean and dry, conventional feeding management is adopted, when the day of delivery is marked as experiment day 0, no feed is fed, the corresponding feed is fed according to groups after delivery, the limit feeding is adopted for 5 days before delivery, the 1 st day is fed for 1.9 kg/day, then 0.5 kg is added every day, free feeding is started on the 6 th day, and free drinking water is adopted.
(1) The production performance of the lactating sow comprises the steps of weighing the birth weight of the piglets immediately after delivery, calculating the birth average weight of the piglets, weighing the weight of each piglet at 08:00 in the morning on the 21 st day of experiment, calculating the weaning average weight of the piglets and the average daily weight gain of the piglets on the 21 st day, and the experimental results are shown in the following table 3.
(2) Daily feed intake of lactating sow daily record of weight change of each experimental sow, calculate average daily feed intake of each sow, average daily feed intake (kg) =total feed intake of group/(experimental days×number of experimental pigs of group), and experimental results are shown in table 3 below.
Table 2 test of stability of Compound acidulants of examples 1 to 5 and comparative examples 1 to 2
TABLE 3 results of experiments on the growth performance of lactating sows
As can be seen from Table 2, the retention rate of the examples in simulated gastric fluid is higher than that of the comparative examples, which shows that the synergy of the wall material cystine-gamma-polyglutamic acid and arginine-chitosan can effectively resist gastric acid, realize low leakage in stomach and accord with pH response mechanism, and the release rate of the examples 1-3 in simulated intestinal fluid is higher, which shows that the wall material can be rapidly dissolved in the neutral environment of intestinal canal and accord with reduction response mechanism. Comparative example 1 and comparative example 2 show that the lack of disulfide cross-linking or arginine modification results in poor stability in gastric juice and reduced intestinal fluid responsiveness.
As shown in Table 3, the results of the sow growth performance test of application examples 1-7 are better than those of comparative examples 1-3, and the compound acidifier provided by the invention is proved to effectively improve the utilization efficiency of the acidifier through an intelligent targeted release mechanism and improve the feeding desire of lactating sows. Meanwhile, weaning average weight of 21-day piglets and average daily weight gain of 21-day piglets of application example 1 and application example 3 are the highest level, and the values are close, so that the feed intake of lactating sows is improved, the lactation amount and the milk quality are improved, and the growth of the piglets is promoted. Meanwhile, the birth weight of piglets has no obvious difference among groups, so that the interference of initial conditions is eliminated, and the experimental result is more comparable.
The test results of comparative application example 1 and comparative application example 2 are inferior to application example 1, and it is proved that cystine-gamma-polyglutamic acid and arginine-chitosan as composite wall materials are two indispensable core elements for achieving the beneficial effects of the present invention, and the absence of either one results in the degradation of product performance. Meanwhile, the results of comparative application examples 1-2 are superior to those of comparative application example 3, and the intelligent microcapsule technology adopted by the invention proves that the traditional acidulant is improved in performance, and the problems of poor targeting property, insufficient stability, single function, loss of active ingredients in the stomach and incapability of acting on the intestinal tract efficiently are solved.
The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.
Claims (10)
1. A compound acidifier for improving the feed intake of lactating sows is characterized by comprising a wall material and a core material, wherein the wall material comprises cystine-gamma-polyglutamic acid and arginine-chitosan, the mass ratio of the cystine-gamma-polyglutamic acid to the arginine-chitosan in the wall material is 1 (0.8-1.2), and the core material comprises organic acid, organic acid salt and fermentation liquor.
2. The compound acidifier for improving the feed intake of the lactating sow according to claim 1, wherein the organic acid comprises one or more of formic acid, fumaric acid, lactic acid, malic acid and benzoic acid, the organic acid salt comprises one or more of ammonium formate, calcium formate and calcium propionate, the fermentation liquor comprises a compound microbial inoculum and a fermentation substrate, the compound microbial inoculum comprises bacillus natto and saccharomycetes, the saccharomycetes comprises one or two of brewer's yeast and candida utilis, and the fermentation substrate comprises 20-30 parts of soybean meal, 1-5 parts of bran, 0.5-2 parts of magnesium sulfate, 2-5 parts of dipotassium hydrogen phosphate, 2-5 parts of yeast extract powder and 60-70 parts of water by weight.
3. The compound acidulant for improving feed intake of lactating sows as claimed in claim 1, wherein the preparation method of the fermentation broth comprises the following steps:
Mixing the bacillus natto liquid and the saccharomycete liquid to prepare a compound microbial agent, inoculating the compound microbial agent into the fermentation substrate for aerobic fermentation, heating the fermentation liquid to terminate all microorganisms and enzyme activities after fermentation, centrifuging to obtain supernatant, and performing aseptic filtration to obtain the fermentation liquid.
4. The compound acidifier for improving the feed intake of the lactating sow according to claim 3, wherein the mixing ratio of bacillus natto liquid to saccharomycete liquid is (1-3): 1, the weight ratio of the compound acidifier to a fermentation substrate is (1-8): 100, and the aerobic fermentation method is to stir and ventilate for 48-72 hours at the temperature of 30-37 ℃.
5. The preparation method of the compound acidifier for improving the feed intake of the lactating sows is characterized by comprising the following steps:
S1, preparing a composite wall material solution and a core material solution, namely respectively dissolving cystine-gamma-polyglutamic acid and arginine-chitosan to prepare solutions, mixing the two solutions to obtain the composite wall material solution, weighing organic acid, organic acid salt and fermentation liquor, mixing the fermentation liquor with the organic acid salt firstly, and then slowly and uniformly mixing the fermentation liquor with the organic acid salt to obtain the core material solution;
S2, complex-phase emulsification and solidification, namely dripping core material solution into corn oil containing a surfactant, homogenizing at high speed under ice bath condition to form primary emulsion, dripping into composite wall material solution, continuously emulsifying, slowly dripping cross-linking agent into the composite emulsion, and stirring;
S3, purifying and collecting, namely centrifuging and washing the mixture obtained in the step S2, dispersing the obtained solid in a freeze-drying protective agent, and obtaining the compound acidifier after freeze-drying and sieving.
6. The method for preparing the compound acidifier for improving the feed intake of the lactating sow as claimed in claim 5, wherein the mass ratio of the organic acid to the organic acid salt to the fermentation liquor is (2-4): 1:1.
7. The method for preparing the compound acidifier for improving the feed intake of the lactating sow as claimed in claim 5, wherein the cross-linking agent is one or two selected from sodium tripolyphosphate and sodium citrate, and the lyoprotectant is one or more selected from trehalose, mannitol and maltodextrin.
8. The compound acidifier for improving the feed intake of the lactating sow according to any one of claims 1 to 4 or the application of the compound acidifier for improving the feed intake of the lactating sow prepared by the preparation method of the compound acidifier for improving the feed intake of the lactating sow according to any one of claims 5 to 7 in the aspect of improving the feed intake of the lactating sow.
9. A method for improving the feed intake of lactating sows is characterized by comprising the following steps of uniformly mixing a compound acidifier with a basic feed to obtain a compound acidifier feed for improving the feed intake of lactating sows, and feeding the compound acidifier feed to lactating sows.
10. The method for increasing the feed intake of a lactating sow of claim 9, wherein the mass ratio of the compound acidulant to the basic diet is (0.5-2) 100.
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