CN102894186B - Albumen powder richening in antimicrobial peptide fly maggot, preparation method for albumen powder and feed additive of albumen powder - Google Patents
Albumen powder richening in antimicrobial peptide fly maggot, preparation method for albumen powder and feed additive of albumen powder Download PDFInfo
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- CN102894186B CN102894186B CN201210385911.1A CN201210385911A CN102894186B CN 102894186 B CN102894186 B CN 102894186B CN 201210385911 A CN201210385911 A CN 201210385911A CN 102894186 B CN102894186 B CN 102894186B
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Abstract
The invention discloses albumen powder richening in antimicrobial peptide fly maggot, a preparation method for the albumen powder and a feed additive of the albumen powder. The albumen powder richening in the antimicrobial peptide fly maggot is prepared by a method comprising the following steps of: (1) feeding flies, luring the flies to lay eggs and feeding larva by using feed after incubation; (2) conducting ultrasound induction on the larva; (3) continuously feeding, collecting the larva, washing, conducting disinfection treatment, and conducting tissue homogenate smashing; and (4) filtering, treating filtrate at 0-5 DEG C, removing surface lipid compositions, drying to obtain the albumen powder; adding auxiliary materials in the albumen powder, pelletizing to obtain the feed additive. A ultrasound stimulation mode under specified conditions is adopted to induce the larva to enable an antimicrobial peptide active ingredient content in a body of a housefly to obviously improve, and the feed additive has certain growth promotion and disease-resistant strengthening effects on litopenaeus vannamei, crucian and the like.
Description
Technical field
The invention belongs to Flyblow albumen powder preparation field,
oneplant and be rich in antibacterial peptide Flyblow albumen powder and preparation method thereof and feed addictive.
Background technology
Along with modern culture fishery scale and intensive development, use green non-pollution feeding additive aquatic animal to obtain people's great attention.In the later stage eighties 20th century, some big cities, the whole nation begin one's study and develop " pollution-free food ".Governments at all levels have formulated corresponding law, rules.After China joined WTO, various places have started " nuisanceless ", " green " aquatic products upsurge.Fish meal is indispensable high-quality protein source in aquatic feeds all the time, but its use exists the problems such as price is high, use amount is large.In addition, in aquaculture, also there is many medication problems, cause the residual of antibiotic, heavy metal etc. in fish body and water body and accumulate, serious impact the development of China's Non-environmental Pollution Aquiculture.Therefore, find that a kind of dietary protein origin is abundant, quality is good, the novel green feed addictive that can reduce or replace antibiotic use has very large Practical significance simultaneously.
Fly-maggot protein is a kind of important protein resource and great exploitation potential for its.A large amount of trophic analysis results show, the rich in protein of fly maggot, and dried protein content is up to 43.62% ~ 55.26%, and various amino acid contents all will be higher than fish meal, and essential amino acids content is 2.3 times of fish meal.In addition, fly maggot has unique immunologic function, and its body contains a kind of " antibacterial active peptide " of very strong bactericidal action, and this active peptide is as long as ten thousand/concentration is just enough to kill the germ of invasion.But the content of this active peptide in fly maggot is very low, in the Flyblow albumen powder therefore preparing, the content of antibacterial active peptide is also very low.At present less to the developmental research of its product, it is only 0.26% that the people such as Lu Jie (housefly maggot antibacterial peptide Study on extraction, 2007, insect journal) utilize heating-chromatography to send out to extract antibacterial peptide extraction yield.
Summary of the invention
In order to overcome in prior art the deficiency of antibacterial peptide content in Flyblow albumen powder, the present invention will solve primary technical problem and be to provide a kind of preparation method who is rich in antibacterial peptide Flyblow albumen powder.
Another technical problem to be solved by this invention be to provide a kind of by the above-mentioned preparation method who is rich in antibacterial peptide Flyblow albumen powder, prepared be rich in antibacterial peptide Flyblow albumen powder.
A technical problem more to be solved by this invention is to provide a kind of feed addictive.
Above-mentioned technical problem to be solved by this invention is achieved by the following technical programs:
Be rich in a preparation method for antibacterial peptide Flyblow albumen powder, comprise the steps: that (1) raise into fly, lure collection to lay eggs, after hatching, with forage feed larva; (2) to larva, carry out supersonic induced; (3) continue to raise 12 ~ 48 hours, collect larva, through cleaning, after disinfecting, carry out tissue homogenate and smash to pieces; (4) filter, filtrate, through 0 ~ 5 ℃ of processing, is removed surperficial lipid component, dry after and get final product;
The described larva of step (2) is 3 instar larvaes; Described supersonic induced of step (2), its condition is: 30 ~ 40 KHzs, process 1 ~ 2 minute 5 ~ 15 minutes, interval, reprocessing 2 ~ 4 times for 40 ~ 60 watts.
Fly of the present invention is preferably housefly, and its formal name used at school is
musca domesticalinnaeus.
As a kind of preferred version: lure collection to lay eggs to be by by the one-tenth fly after mating, to put into collection ovum cup described in step (1), what lure collection to lay eggs to carry out under the effect of material; The described collection material of laying eggs that lures is: wheat bran+pig blood+0.01 % by weight ammoniacal liquor.
As a kind of preferred version: the feed described in step (1) is wheat bran+pig blood+yeast.
As a kind of preferred version: supersonic induced described in step (2), condition is: 35 KHzs, process 1 minute 10 minutes, interval, reprocessing 3 times for 50 watts.
As a kind of preferred version: the feeding time described in step (3) is 24 hours.Collecting larva can utilize negative phototaxis to collect.
As a kind of preferred version: the 80 order gauzes that are filtered into described in step (4) filter; Filtrate is through 0 ℃ of processing.
A kind of by above-mentioned preparation method, prepared be rich in antibacterial peptide Flyblow albumen powder.
A feed addictive, comprises antibacterial peptide Flyblow albumen powder and the auxiliary material of being rich in of the present invention.
Described Optimization of Adjuvant is cornstarch.
Described cornstarch, its content is 5% of additive gross weight.
The preparation method of feed addictive of the present invention, is prepared by following steps: by the antibacterial peptide Flyblow albumen powder that is rich in of the present invention, add auxiliary material, granulate and get final product.
As a kind of preferred version: a kind of preparation method of feed addictive, is prepared by following steps: by the antibacterial peptide Flyblow albumen powder that is rich in of the present invention, add the cornstarch of 5 % by weight to carry out cold granulation, be processed into granular pattern additive.
In feed addictive application aquaculture of the present invention.Can promote aquatic products growth, strengthen aquatic products resistance against diseases.
Beneficial effect of the present invention: (1) the present invention adopts the ultrasound stimulation mode under specified conditions to induce larva, significantly raises little molecule antibacterial peptide active component content in housefly body; (2) supersonic induced method used in the present invention is simple, the large-scale production that is suitable for being rich in antimicrobial peptide protein powder; (3) in feed, adding 15 % by weight feed addictives has certain growth promoting function to Environment of Litopenaeus vannamei Low, crucian etc., and its meat improves, and delicate flavour amino acid content raises, and can not cause new pollution to breeding water body; (4) experiment in vitro result shows, feed addictive of the present invention all has certain inhibitory action to Pseudomonas fluorescens, Vibrio anguillarum, Aeromonas caviae, streptococcus etc.; In body, experimental result shows, the fish that the pathogenic microorganisms such as Pseudomonas fluorescens, Vibrio anguillarum are caused and shrimp body infect and have certain preventive and therapeutic effect.
Accompanying drawing explanation
Fig. 1: the preparation method's process chart that is rich in antibacterial peptide Flyblow albumen powder and feed addictive;
Fig. 2: the affect figure of feed addictive on water body PH, salinity, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity.
The specific embodiment
Below in conjunction with specific embodiment, further explain the present invention, but embodiment does not limit in any form to inventing itself.
Embodiment 1 is rich in the preparation of antibacterial peptide Flyblow albumen powder and feed addictive
Housefly derives from disease prevention and control center, Guangdong Province, and through long-time acclimation.After becoming fly mating, put into collection ovum cup and (lure and collect the material of laying eggs: wheat bran+pig blood+0.01% ammoniacal liquor), collect fly blow, after hatching, proceed to basal feed (wheat bran+pig blood+yeast) and feed; Larvae development to 3 age, to carry out supersonic inducedly, condition is as follows: 35 KHzs, process 1 minute 10 minutes, interval, reprocessing 3 times for 50 watts; Continue to raise 24 hours, utilize negative phototaxis to collect larva; After cleaning, disinfecting, carry out tissue homogenate and smash to pieces; 80 order gauzes filter, and filtrate, through 0 degree Celsius of low temperature pretreatment 40 minutes, is removed surperficial lipid component; Spraying is so dry that to be rich in antibacterial peptide Flyblow albumen powder; Then add appropriate amount of auxiliary materials (cornstarch 5 % by weight) to carry out cold granulation, be processed into granular pattern feed addictive.The feed addictive that the present embodiment is obtained carries out the experiment shown in embodiment 3 ~ 6, the results are shown in specific embodiment.
Embodiment 2
By the present invention through specific ultrasonic (35 KHzs, process 1 minute for 50 watts, 10 minutes, interval, reprocessing 3 times) induction process obtain after common house-fly grub be rich in antibacterial peptide Flyblow albumen powder, with the Flyblow albumen powder obtaining without supersonic induced treatment step, through dextran gel filtration, with the composition of BCA method detection molecules amount < 30KD, and measure its bacteriostatic activity respectively.Result demonstration is after induction is processed, and the ratio that the composition of < 30KD accounts for Flyblow albumen powder rises to (21.33 ± 1.22) % by weight by original (Flyblow albumen powder obtaining without special ultrasonic induction larva step) (8.92 ± 0.13) % by weight.The method that adopts drug susceptability test paper to detect is tested, the composition of finding molecular weight < 30KD has obvious antibacterial action to EHEC, staphylococcus aureus, helicobacter pylori etc. when concentration is 40 μ g/ml, inhibiting rate is respectively (82.43 ± 2.53) %, (57.23 ± 3.21) %, (45.66 ± 1.87) %, experimental results show that the content of antibacterial active peptide (composition of molecular weight < 30KD) improves greatly.
The impact of embodiment 3 feed addictives on nuisanceless sea-farming Growth of Litopenaeus vannamei performance and water quality
With 5 % by weight (A group), the feed addictive of 10 % by weight (B group) and 15 % by weight (C group) substitutes the fish meal in D group respectively, and D group is basal feed control group (containing fish meal 15 % by weight).With above-mentioned 4 kinds of feeds, carried out a culture experiment of 8 weeks by a definite date.Throw something and feed every day feed 4 times of duration of test.When culture experiment finishes, calculate respectively that body is long, total length and the growth performance such as carapace is wide.The body of experimental group C is long, and the wide growth of total length and carapace is all greater than control group D, and the body of experimental group C is long, and the wide growth of total length and carapace is all greater than experimental group A and B.The survival rate that ABCD respectively organizes prawn is respectively 88.0%, 90.3%, 96.0% and 90.0%, AB group basically identical with the survival rate of control group prawn, and C group has significant difference with the survival rate of control group prawn, experimental group is significantly higher than control group (P <0.05).This can illustrate and use appropriate feed addictive Peru Fish Dietary to raise the survival rate that prawn can significantly improve prawn.Experimental group and control group prawn growing state are in Table 1.
By detecting the PH, salinity, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity etc. of breeding water body, find to use three groups of feed addictive, water quality situation not have larger difference with control group, illustrate to use the feed addictive can be to water quality increase pollution.Concrete data are shown in accompanying drawing 2.
The impact of embodiment 3 feed addictives on nuisanceless freshwater aquiculture crucian growth performance and water quality
Young crucian, purchased from Foshan aquatic farm, is chosen same batch, healthy fry (specification is 50～70g), carries out 12 weeks culture experiments.Wherein, 1 group is control group, the basal feed of throwing something and feeding (containing fish meal); All the other 3 groups is test group, contains the test feed of feed addictive 5 % by weight, 10 % by weight and 15 % by weight in the basal feed of throwing something and feeding respectively.By additive, the impact effect of growth is measured, found that 10 % by weight additive groups have the highest the relative weight gain rate and specific growth rate, there is minimum feed coefficient.The fish meal substituting in feed with additive carries out 12 weeks culture experiments, can significantly improve the relative weight gain rate and the specific growth rate of crucian, reduce feed coefficient, strengthen to a certain extent the digestive enzyme activity of intestines and liver pancreas, improve feed efficiency, and improve to a certain extent muscle containing meat rate.In addition, this additive can significantly improve neutrophil leucocyte sterilizing function, significantly promotes the generation of active oxygen to improve respiratory burst ability, and can significantly improve lysozyme activity, superoxide dismutase activity and TAC, strengthens fish body resistance against diseases.And in the flesh of fish, the content of total free amino acid and delicate flavour amino acid (mainly comprising aspartic acid, glutamic acid, glycine and alanine) raises, and mouthfeel Fresh & Tender in Texture is good.Concrete data are in Table 2～6.
By detecting the PH, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity etc. of breeding water body, find to use three groups of fly maggot powder, water quality situation not have larger difference with control group, illustrate to use the fly maggot powder additive can be to water quality increase pollution.
Embodiment 4 feed addictives infect the impact of Growth of Litopenaeus vannamei on Vibrio anguillarum
In water body, inoculate into Vibrio anguillarum, inoculum density is 120cfu/ml, the feed that control group fed contains 15 % by weight fish meal, the feed that experimental group is fed and contained 15 % by weight additives.After 3 weeks, control group prawn hypoergia, anorexia, individuality is become thin, and most prawn appendages redden, and the death rate is (56.22 ± 1.49) %, and microscopic examination finds that its hemolymph is swollen a large amount of bacteriums; The negligible amounts that additive is fed the ill shrimp of experimental group, symptom is lighter, and the death rate is (7.92 ± 0.71) %.
Embodiment 5 feed addictives infect the impact of crucian growth on Pseudomonas fluorescens
Adopt the mode of lumbar injection to infect crucian, inoculum concentration is 1.5 * 10
5cfu/ml, the feed that control group fed contains 15 % by weight fish meal, the feed that experimental group is fed and contained 15 % by weight additives.Control group is after infecting 4 days, and the minimizing of moving about, infected after 20 days, and the death rate reaches (89.35 ± 2.88) %, shows as body surface hyperemia, and hemorrhage, scale outwards opens, and becomes flexible and easily comes off, and has red ascites in abdominal cavity, visible a large amount of bacteriums under mirror.The additive experimental group of feeding, the growth performance that infects fish increases, and the death rate is reduced to (25.65 ± 1.48) %.
Embodiment 6 feed addictive water-soluble substanceses are the inhibitory action to Vibrio anguillarum, Pseudomonas fluorescens, Aeromonas caviae, streptococcus, fish lactobacillus etc. in vitro.
Appropriate additive and water mix, and centrifugal 10 minutes of 3000r/min, gets supernatant, vacuum drying.Measure its external minimal inhibitory concentration to Vibrio anguillarum, Pseudomonas fluorescens, Aeromonas caviae, streptococcus etc.With LB fluid nutrient medium, dried material doubling dilution is diluted 10 times altogether, each dilution is 1ml, adjustment cell concentration is 105CFU/ml, in 37 ℃, 150r/min, cultivate 18～20h by the colibacillary growing state of colorimetric estimation, in conjunction with probity regression analysis, obtain the repressed concentration of 80% bacterium and be minimal inhibitory concentration (minimal inhibitory concentrations, MIC).Result is as shown in table 7:
Claims (7)
1. be rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that, described in be rich in antibacterial peptide Flyblow albumen powder preparation method comprise: (1) raises into fly, lures collection to lay eggs, after hatching, with forage feed larva; (2) to larva, carry out supersonic induced; (3) continue to raise 12 ~ 48 hours, collect larva, through cleaning, after disinfecting, carry out tissue homogenate and smash to pieces; (4) filter, filtrate, through 0 ~ 5 ℃ of processing, is removed surperficial lipid component, dry after and get final product;
The described larva of step (2) is 3 instar larvaes; Described supersonic induced of step (2), its condition is: 30 ~ 40 KHzs, process 1 ~ 2 minute 5 ~ 15 minutes, interval, reprocessing 2 ~ 4 times for 40 ~ 60 watts;
Described in step (1), lure collection to lay eggs to be by by the one-tenth fly after mating, to put into collection ovum cup, what lure collection to lay eggs to carry out under the effect of material; The described collection material of laying eggs that lures is: wheat bran+pig blood+0.01 % by weight ammoniacal liquor.
Feed described in step (1) is wheat bran+pig blood+yeast.
2. be according to claim 1ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that supersonic induced described in step (2), condition is: 35 KHzs, process 1 minute 10 minutes, interval, reprocessing 3 times for 50 watts.
3. be according to claim 1ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that the feeding time described in step (3) is 24 hours.
4. be according to claim 1ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that the 80 order gauzes that are filtered into described in step (4) filter; Filtrate is through 0 ℃ of processing.
5. be according to claim 1ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that described feed addictive comprises to be rich in antibacterial peptide Flyblow albumen powder described in auxiliary material and claim 1-5 any one.
6. be according to claim 5ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that described auxiliary material is cornstarch.
7. be according to claim 6ly rich in the application of antibacterial peptide Flyblow albumen powder in feed addictive, it is characterized in that described cornstarch, its content is 5% of feed addictive gross weight.
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| CN104137943A (en) * | 2013-05-07 | 2014-11-12 | 王虎生 | A continuous production method of high-activity fly maggot protein powder |
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| CN105519823A (en) * | 2016-01-21 | 2016-04-27 | 厦门蓝鳍海洋生物科技有限公司 | Special fly maggot immune feed for aquatic products and preparation method thereof |
| CN106432404A (en) * | 2016-10-25 | 2017-02-22 | 蚌埠学院 | Method for separating antibacterial peptides |
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| CN114097711B (en) * | 2021-11-23 | 2023-04-25 | 华南理工大学 | Method for inducing yellow meal worm to produce antibacterial peptide, preparation method and application of yellow meal worm antibacterial peptide |
| CN114836497B (en) * | 2022-05-21 | 2023-04-18 | 广东日和堂医药科技有限公司 | Preparation method of maggot antibacterial peptide, composition containing maggot antibacterial peptide and preparation method of composition |
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