CN102894186A - 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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- CN102894186A CN102894186A CN2012103859111A CN201210385911A CN102894186A CN 102894186 A CN102894186 A CN 102894186A CN 2012103859111 A CN2012103859111 A CN 2012103859111A CN 201210385911 A CN201210385911 A CN 201210385911A CN 102894186 A CN102894186 A CN 102894186A
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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 the 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 the 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 the China joined WTO, various places have started " nuisanceless ", " green " aquatic products upsurge.Fish meal is indispensable high-quality protein source in the aquatic feeds all the time, but there are the problems such as price is high, use amount is large in its use.In addition, also there are many medication problems in the aquaculture, cause the residual of antibiotic, heavy metal etc. in fish body and the water body and accumulate, seriously influenced the development of China's Non-environmental Pollution Aquiculture.Therefore, seek that a kind of dietary protein origin is abundant, quality is good, the novel green feed addictive that can reduce or replace simultaneously the antibiotic use has very large Practical significance.
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 just is enough to kill the germ of invasion.But the content of this active peptide in fly maggot is very low, and the content of antibacterial active peptide is also very low in the Flyblow albumen powder that therefore prepares.At present less to the developmental research of its product, it only is 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 the prior art deficiency of antibacterial peptide content in the Flyblow albumen powder, the present invention will solve primary technical problem and be to provide a kind of preparation method who is rich in the antibacterial peptide Flyblow albumen powder.
Another technical problem to be solved by this invention is to provide a kind of and is rich in the antibacterial peptide Flyblow albumen powder by what the above-mentioned preparation method who is rich in the antibacterial peptide Flyblow albumen powder prepared.
An again technical problem 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:
A kind of preparation method who is rich in the antibacterial peptide Flyblow albumen powder comprises the steps: that (1) raise into fly, lures collection to lay eggs, after the hatching, with the forage feed larva; (2) carry out supersonic induced to larva; (3) continue to raise 12 ~ 48 hours, collect larva, through cleaning, after disinfecting, carry out tissue homogenate and smash to pieces; (4) filter, filtrate is removed surperficial lipid component through 0 ~ 5 ℃ of processing, after the drying and get final product;
The described larva of step (2) is 3 instar larvaes; Step (2) is described supersonic induced, and its condition is: 30 ~ 40 KHzs, processed interval 5 ~ 15 minutes, reprocessing 2 ~ 4 times 1 ~ 2 minute 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 with the one-tenth fly after the mating, to put into collection ovum cup described in the 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 the step (1) is wheat bran+pig blood+yeast.
As a kind of preferred version: supersonic induced described in the step (2), condition is: 35 KHzs, processed interval 10 minutes, reprocessing 3 times 1 minute for 50 watts.
As a kind of preferred version: the feeding time described in the 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 the step (4) filter; Filtrate is through 0 ℃ of processing.
A kind ofly be rich in the antibacterial peptide Flyblow albumen powder by what above-mentioned preparation method prepared.
A kind of 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 are 5% of additive gross weight.
The preparation method of feed addictive of the present invention is prepared by following steps: with 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: with the antibacterial peptide Flyblow albumen powder that is rich in of the present invention, the cornstarch that adds 5 % by weight carries out cold granulation, is processed into the granular pattern additive.
Feed addictive of the present invention is used in the aquaculture.Can promote the aquatic products growth, strengthen the aquatic products resistance against diseases.
Beneficial effect of the present invention: (1) the present invention adopts the ultrasound stimulation mode under the specified conditions that larva is induced, so that little molecule antibacterial peptide active component content significantly raises in the housefly body; (2) supersonic induced method used in the present invention is simple, the large-scale production that is suitable for being rich in the antimicrobial peptide protein powder; (3) adding 15 % by weight feed addictives in the feed has certain growth promoting function to Environment of Litopenaeus vannamei Low, crucian etc., and its meat improves, and the delicate flavour amino acid content raises, and can not cause new pollution to breeding water body; (4) the 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.; Experimental result shows in the body, and fish and the infection of shrimp body that the pathogenic microorganisms such as Pseudomonas fluorescens, Vibrio anguillarum are caused have certain preventive and therapeutic effect.
Description of drawings
Fig. 1: the preparation method's process chart that is rich in antibacterial peptide Flyblow albumen powder and feed addictive;
Fig. 2: feed addictive is on the figure that affects of water body PH, salinity, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity.
The specific embodiment
Further explain the present invention below in conjunction with specific embodiment, but embodiment does not do any type of restriction 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 the 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 the hatching, change basal feed (wheat bran+pig blood+yeast) over to and feed; Larvae development to 3 age, to carry out supersonic inducedly, condition is as follows: 35 KHzs, processed interval 10 minutes, reprocessing 3 times 1 minute 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 is removed surperficial lipid component through 0 degree centigrade of low temperature preliminary treatment 40 minutes; Spray-drying must be rich in the antibacterial peptide Flyblow albumen powder; Then add appropriate amount of auxiliary materials (cornstarch 5 % by weight) and carry out cold granulation, be processed into the granular pattern feed addictive.The feed addictive that the present embodiment is obtained carries out the experiment shown in the embodiment 3 ~ 6, the results are shown in specific embodiment.
Embodiment 2
With the present invention through specific ultrasonic (35 KHzs, processed 1 minute for 50 watts, interval 10 minutes, reprocessing 3 times) induce process obtain behind the common house-fly grub be rich in the antibacterial peptide Flyblow albumen powder, with the Flyblow albumen powder that obtains 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.The result shows after inducing processing, 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 of inducing the larva step to obtain without special ultrasonic) (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.
Embodiment 3 feed addictives are on the impact of 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 the D group respectively, and the D group is basal feed control group (containing fish meal 15 % by weight).Carried out a by a definite date culture experiment in 8 weeks with above-mentioned 4 kinds of feeds.Throw something and feed every day feed 4 times of duration of test.The growth performance such as when culture experiment finishes, calculate respectively that body is long, total length and carapace are 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 the survival rate of C group and control group prawn has significant difference, and experimental group is significantly higher than control group (P＜0.05).This can illustrate and use an amount of feed addictive Peru Fish Dietary to raise the survival rate that prawn can significantly improve prawn.Experimental group and control group prawn growing state see Table 1.
PH by detecting breeding water body, salinity, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity etc. find to use three groups of feed addictive, and the water quality situation not have larger difference with control group, illustrates to use the feed addictive can be to water quality increase pollution.Concrete data are seen accompanying drawing 2.
Embodiment 3 feed addictives are on the impact of nuisanceless freshwater aquiculture crucian growth performance and water quality
Young crucian is available from the Foshan aquatic farm, chooses same batch, (specification is 50～70g) to healthy fry, carries out 12 all 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.To the affects on the growth effect measuring, find that 10 % by weight additive groups namely have the highest the relative weight gain rate and specific growth rate by additive, have minimum feed coefficient.The fish meal that substitutes in the feed with additive carries out 12 all 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 contain the meat rate.In addition, this additive can significantly improve the neutrophil leucocyte sterilizing function, significantly promotes the generation of active oxygen with raising respiratory burst ability, and can significantly improve lysozyme activity, superoxide dismutase activity and TAC, strengthens fish body resistance against diseases.And the content of total free amino acid and delicate flavour amino acid (mainly comprising aspartic acid, glutamic acid, glycine and alanine) raises in the flesh of fish, and mouthfeel Fresh ﹠ Tender in Texture is good.Concrete data see Table 2～6.
PH by detecting breeding water body, ammonia-nitrogen content, nitrous acid nitrogen, total alkalinity etc. find to use three groups of fly maggot powder, and the water quality situation not have larger difference with control group, illustrates 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
Inoculation enters Vibrio anguillarum in the water body, and inoculum density is 120cfu/ml, and control group fed contains the feed of 15 % by weight fish meal, and experimental group is fed and contained the feed of 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 crucian affects on the growth to Pseudomonas fluorescens
Adopt the mode of lumbar injection to infect crucian, inoculum concentration is 1.5 * 10
5Cfu/ml, control group fed contain the feed of 15 % by weight fish meal, and experimental group is fed and contained the feed of 15 % by weight additives.Control group is after infecting 4 days, and the minimizing of moving about was 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 easily to come off, and red ascites is arranged in the abdominal cavity, visible a large amount of bacteriums under the 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 in external inhibitory action to Vibrio anguillarum, Pseudomonas fluorescens, Aeromonas caviae, streptococcus, fish lactobacillus etc.
An amount of additive and water mixing, 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 the LB fluid nutrient medium dried material doubling dilution is diluted 10 times altogether, each dilution is 1ml, the adjustment cell concentration is 105CFU/ml, cultivate 18～20h by the colibacillary growing state of colorimetric estimation in 37 ℃, 150r/min, in conjunction with the probity regression analysis, obtain the repressed concentration of 80% bacterium and be minimal inhibitory concentration (minimal inhibitory concentrations, MIC).The result is as shown in table 7:
Claims (10)
1. preparation method who is rich in the antibacterial peptide Flyblow albumen powder is characterized in that luring collection to lay eggs, after the hatching, with the forage feed larva by comprising the steps: that (1) raise into fly; (2) carry out supersonic induced to larva; (3) continue to raise 12 ~ 48 hours, collect larva, through cleaning, after disinfecting, carry out tissue homogenate and smash to pieces; (4) filter, filtrate is removed surperficial lipid component through 0 ~ 5 ℃ of processing, after the drying and get final product;
The described larva of step (2) is 3 instar larvaes; Step (2) is described supersonic induced, and its condition is: 30 ~ 40 KHzs, processed interval 5 ~ 15 minutes, reprocessing 2 ~ 4 times 1 ~ 2 minute for 40 ~ 60 watts.
2. the preparation method who is rich in the antibacterial peptide Flyblow albumen powder according to claim 1 is characterized in that luring collection to lay eggs to be by with the one-tenth fly after the mating, to put into collection ovum cup described in the 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.
3. the preparation method who is rich in the antibacterial peptide Flyblow albumen powder according to claim 1 is characterized in that the feed described in the step (1) is wheat bran+pig blood+yeast.
4. the preparation method who is rich in the antibacterial peptide Flyblow albumen powder according to claim 1 is characterized in that supersonic induced described in the step (2), and condition is: 35 KHzs, processed interval 10 minutes, reprocessing 3 times 1 minute for 50 watts.
5. the preparation method who is rich in the antibacterial peptide Flyblow albumen powder according to claim 1 is characterized in that the feeding time described in the step (3) is 24 hours.
6. the preparation method who is rich in the antibacterial peptide Flyblow albumen powder according to claim 1 is characterized in that the 80 order gauzes that are filtered into described in the step (4) filter; Filtrate is through 0 ℃ of processing.
7. be rich in the antibacterial peptide Flyblow albumen powder by what each described preparation method who is rich in the antibacterial peptide Flyblow albumen powder of claim 1～6 prepared.
8. a feed addictive is characterized in that comprising antibacterial peptide Flyblow albumen powder and the auxiliary material of being rich in claimed in claim 7.
9. feed addictive according to claim 8 is characterized in that described auxiliary material is cornstarch.
10. feed addictive according to claim 9 is characterized in that described cornstarch, and its content is 5% of feed addictive gross weight.
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| CN103583910A (en) * | 2013-11-19 | 2014-02-19 | 湛江恒兴珊瑚饲料有限公司 | Compound feed capable of improving flavor of trachinotus ovatus meat and preparation method of compound feed |
| CN104137943A (en) * | 2013-05-07 | 2014-11-12 | 王虎生 | A continuous production method of high-activity fly maggot protein powder |
| 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 |
| CN105614074A (en) * | 2015-12-25 | 2016-06-01 | 广东鑫肽生物科技股份有限公司 | Novel insect protein concentrated feed with additive amount being 4% |
| CN106432404A (en) * | 2016-10-25 | 2017-02-22 | 蚌埠学院 | Method for separating antibacterial peptides |
| CN107494893A (en) * | 2017-07-27 | 2017-12-22 | 珠海天凯生物科技有限公司 | A kind of feed additive and a kind of premixed feed and preparation method thereof |
| CN108041346A (en) * | 2017-12-11 | 2018-05-18 | 中山大学 | A kind of America elver fly-maggot protein fodder additive and its preparation method and application of disease-resistant growth-promoting |
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| CN114097711A (en) * | 2021-11-23 | 2022-03-01 | 华南理工大学 | Method for inducing yellow mealworms to produce antibacterial peptide, preparation method and application of yellow mealworms antibacterial peptide |
| CN114836497A (en) * | 2022-05-21 | 2022-08-02 | 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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| CN104137943A (en) * | 2013-05-07 | 2014-11-12 | 王虎生 | A continuous production method of high-activity fly maggot protein powder |
| CN103385891A (en) * | 2013-07-02 | 2013-11-13 | 金陵科技学院 | Application of maggot extract in preparing medicament that enhances immunity of inactivated vaccine for pigeon Newcastle disease |
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| CN103583910B (en) * | 2013-11-19 | 2014-12-10 | 湛江恒兴珊瑚饲料有限公司 | Compound feed capable of improving flavor of trachinotus ovatus meat and preparation method of compound feed |
| CN105614074A (en) * | 2015-12-25 | 2016-06-01 | 广东鑫肽生物科技股份有限公司 | Novel insect protein concentrated feed with additive amount being 4% |
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| CN114097711A (en) * | 2021-11-23 | 2022-03-01 | 华南理工大学 | Method for inducing yellow mealworms to produce antibacterial peptide, preparation method and application of yellow mealworms antibacterial peptide |
| 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 |
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