CN1067887C - New drug for preventing and curing prawn disease and its manufacturing method - Google Patents
New drug for preventing and curing prawn disease and its manufacturing method Download PDFInfo
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- CN1067887C CN1067887C CN97106607A CN97106607A CN1067887C CN 1067887 C CN1067887 C CN 1067887C CN 97106607 A CN97106607 A CN 97106607A CN 97106607 A CN97106607 A CN 97106607A CN 1067887 C CN1067887 C CN 1067887C
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Abstract
The present invention relates to novel method for preventing and treating prawn diseases. The present invention comprises pokeberry root, lysozyme, garlic oil, cilijing and gynostemma pentaphylla. The present invention has the broad-spectrum killing effect on various viruses and various bacteria, and simultaneously, the present invention enhances the immunity, the disease resistance and the survival rate of shrimps, and overcomes the defect of antivirus capability deficiency (the shrimp medicine in the prior art only has the bacterium resistance capability).
Description
The present invention relates to the control of shrimp disease, is a kind of novel shrimp medicine of preventing and treating epiphytotics generation of prawn and control epidemic situation.
China's prawn culturing development is swift and violent, has now become the big product of the first in the world shrimp state and maximum row shrimp exported country.But because the outburst of prawn disease and popular makes shrimp culture industry suffer huge strike and loss.Though have hundreds of preparation medicine selling use now, effect is not satisfactory, wherein some still uses antibiotic in a large number, this not only makes pathogenic bacteria produce drug resistance and Drug resistance, and residual in a large number in the shrimp body, cause the further deterioration of shrimp body and environment, produce vicious cycle.Some imported medicine also cannot be accepted by the vast shrimp farming of China owing to price factor.Find that after deliberation the epidemic diseases of prawn largely is to be caused by various antibacterials certainly, but mainly still cause by multiple virus, so, use antibiotic medicine to can't resolve problem purely.Cause that the popular main virus of prawn disease has: 1, leukasmus rhabdovirus (WSBV); 2, the tiny sample virus of hepatopancrease (HPV); 3, infectiousness is subcutaneous and hematopoietic tissue necrosis is viral (IHHNY); 4, yellow head rod shape virus (YHV); 5, MBV (MBV); 6, the downright bad baculovirus of mid intestinal gland (BMN) waits more than ten to plant virus, and wherein WSBV harm is maximum, infects all cultured prawns, causes general property epidemic diseases, and present serious " leukoderma " caused by WSBV.The explosive epidemic diseases of prawn, the reason of searching to the bottom mainly is by due to biological causing a disease (virus, antibacterial, fungus, the parasitic sexually transmitted disease (STD) of protozoacide) and pathogenic (diseases such as environment, trophism, physiological) the co-infected shrimp body initiation of inanimate.
The objective of the invention is: from biological morbific angle, providing a kind of can prevent and treat and can carry out the new drug that shrimp autoimmunity, premunition and survival rate were killed and improved to broad spectrum activity to various viruses bacillary prawn disease, thereby control for the situation that China prawn culturing production is glided, fundamentally change China the state that the control of shrimp disease lacks active drug is made contributions.
Task of the present invention is achieved in that
Test after deliberation, we select for use lysozyme, these two kinds of compositions of Radix Phytolaccae as necessary composition, be aided with again that Bulbus Allii is disorderly, thorn boule, these three kinds of compositions of Herb Gynostemmae Pentaphylli are made a kind of new drug that includes above-mentioned five kinds of compatibilities, the task that this new drug can ideally be finished the object of the invention and proposed.Why select this its pharmacology principle of five kinds of compositions as follows:
The Phytolaccaceae plant pokeweed comprises domestic and international various Radix Phytolaccaes (Phytolacca) such as pokeroot, multiple medicines Radix Phytolaccae, shows according to modern pharmacological research, and the active ingredient of Radix Phytolaccae class Chinese medicine is fixed Saponin, polysaccharide and histamine etc.Wherein the latter is considered to one of toxicity or zest composition, because the whole plant of Radix Phytolaccae is distributed with pokeweed antiviral protein, and these antiviral proteins (PAP) have very strong cytotoxicity and broad-spectrum disease resistance cytotoxic activity, it is synthetic to hinder the translation of meeting virus effectively, has the broad-spectrum antivirus action.The antiviral function of pokeweed antiviral protein is based on basic RNAN-glycosidase activity.It is generally acknowledged that after cell is by viral infection some variations can take place the plasma membrane of cell, make pokeweed antiviral protein to enter at an easy rate in the cell, kill by the cell of viral infection.Interrupt the amplification of virus in the cell, stop the further infection of virus, reach antiviral effect.Pokeweed antiviral protein has broad spectrum activity to the resistance of virus, and it not only has resistance to plant virus, and animal RNA and DNA viruses are also had resistance.
Lysozyme (Lysozyme) is not only to the G+ bacterium, and to G-such as escherichia coli, Pullorum Disease salmonella, Bacillus proteus, aerobacteria, Salmonella typhimurium etc., to fungus such as Mucor, aspergillosis, penicillium sp etc., stronger bacteriolysis arranged all.
Bulbus Allii is used as medicine with bulb, and its Main Ingredients and Appearance is a garlicin, includes the sulfide compound of multiple pi-allyl and methyl composition etc., and its main pharmacological is an antibacterial action.To various pathogens, as iron link chain coccus, vibrio cholera etc. tangible bacteriostasis and bactericidal action are arranged, still responsive to some antibiotics fastbacteria.Bulbus Allii has antifungic action simultaneously, and multiple pathomycete is comprised that Candida albicans has inhibition or killing action.
Fructus Rosae Normalis is used as medicine with fruit, contain rich in amino acid, vitamin, trace element and enzyme, wherein ascorbic content is abundant especially, and pressing filter fruit juice actual measurement Vc is 1500-2500mg/100ml, Vb about 2.4%, Vp is about 2.8%, and its pharmacological effect improves the content of SOD for improving the immunity of body, eliminate the chain reaction of free radical, stop the generation of peroxide lipid (LPO), delaying cell aging, and functions such as digestion-promoting spleen-invigorating astringing to arrest diarrhea, expelling summer-heat are arranged.
Herb Gynostemmae Pentaphylli is with stem people medicine, and it has the active ingredient of Radix Ginseng, is described as " southern Radix Ginseng ", and its Main Ingredients and Appearance of Herb Gynostemmae Pentaphylli is multiple Saponin, and pharmacology test and clinical research show to have enhance immunity, the anti-ageing function of waiting for a long time.
We use the modern biotechnology means with above Chinese medicine of the five flavours material, keep bioactive substance, and reasonable compatibility is succeeded in developing shrimp medicine of the present invention.
Below introduce the method for making of medicine of the present invention:
In five kinds of compatibilities of shrimp medicine of the present invention, lysozyme, Radix Phytolaccae powder are necessary compositions, the mixed seed shrimp medicine that can meet the object of the invention that just can be mixed with of these two kinds of compositions.The garlicin of thorn boule and Herb Gynostemmae Pentaphylli mix the autoimmunity and the resistance against diseases that can further strengthen prawn, play assosting effect, the medical material that increases some other useful and harmless compositions certainly more also is feasible scheme.Above-mentioned these five kinds of compatibilities all given make earlier Powdered, again formula proportion in accordance with regulations mix stir shrimp medicine of the present invention.Powdered shrimp medicine of the present invention cultured in the pond with the mixed graininess input that forms a grain of the further mix of shrimp bait again use.About the manufacturing of shrimp medicine, the powder that will prepare five kinds of compositions is as intermediate raw material earlier, and wherein (1) lysozyme is that commercially available commodity are sold (its commodity are called the FE lysozyme) by Fudan University's school of life and health sciences manufacturing.It is a lenticular, from albumen, extract, regulate the albumen pH value, spent ion exchange resin absorption, extract and get, have the various character of antibiotics, molecular weight 14.307, alkaline protein, isoelectric point, IP PH10.5～11.0, being soluble in saline, is a kind of off-white color crystalloid powder, odorless, optimum PH6.6, very stable in acid solution, can be heat-resisting under acetone and ethanol effect, generating precipitation, solubilized bacterial cell more than 553, particularly micrococcus is had and pretend usefulness, this is because the polysaccharide of the glycoprotein of its pair cell membranous wall has decomposition, can decompose chitin and ethylene glycol chitin, obtains carbonate.Hydrochlorate and nitrate crystallization etc.(2) preparation process of Radix Phytolaccae powder is: adopt the Phytolaccaceae plant pokeweed, comprise domestic and international various Radix Phytolaccaes (Phytolacca) Herb or parts such as pokeroot, multiple medicines Radix Phytolaccae, at first bright Radix Phytolaccae is cleaned, cutting, drying, pulverizing, sieves at last, granularity is controlled at about (80) order, concrete operations require: will pluck the bright Radix Phytolaccae of June to JIUYUE as raw material, cut into 2～3cm section material, under 50 ± 9 ℃ temperature, dry or dry, to water content be about 8%, pulverizing is crossed 50 orders～80 mesh sieves and is promptly got satisfactory Radix Phytolaccae powder.As the substituent of Radix Phytolaccae powder, also can use various " pokeweed antiviral proteins " of the method acquisition of utilization protein separation is these six kinds of PAP, PAP II, PAP-R, PAP-S, PAP-C and aPAP.(3) preparation of garlicin beta-schardinger dextrin-inclusion compound: get in the water that beta-schardinger dextrin-(general chemical reagent shop sale is all arranged) is dissolved in 12 times of weight, temperature is controlled at 20～30 ℃, drip and amplify Bulbus Allii oil, constantly stirred five hours, standing over night, to doing, under 50 ℃ temperature, dry, be crushed to 80 orders with the vacuum pump sucking filtration.(4) preparation of thorn boule: the Fructus Rosae Normalis natural juice (commercially available) of measuring by weight 40% adds 60% medical starch (commercially available) and puts into mixer and make soft material, granulates with 24 purpose screen clothes, dry below 50 ℃, gets 24 purpose powder, is crushed to 80 orders again.(5) the Herb Gynostemmae Pentaphylli powder is the (place of production: Hubei Province Shennongjia medicated tea medicated diet factory), commercial goods.
Just the ratio in following provisions mixes stirring after above five kinds of powder requirements in accordance with regulations as intermediate raw material prepare, ingredient proportion by weight percentage:
| The Radix Phytolaccae powder | 34.71～50.14％ |
| Lysozyme | 7.12～8.70％ |
| Garlicin β-cyclodextrin inclusion compound | 3.03～4.38％ |
| The thorn boule | 37.61～53.04％ |
| The Herb Gynostemmae Pentaphylli powder | 1.54～2.32％ |
After finishing prescription and stirring,, can bottle and pack even if the preparation of shrimp medicine of the present invention is finished.
The use of shrimp medicine of the present invention: the dosage of (1) shrimp medicine of the present invention is 1% of a dried bait, and as one mu of one season of shrimp pond total consumption bait 50kg (100 jin, a traditional unit of weight), then consuming the medicine total amount is 0.5kg (promptly one bottle).Under the normal condition, offer medicine every day 2 times, period of disease should increase dose, and promptly 2% of dried bait.(2) bait of the present invention and dried granular bait for fish is bonding: calculate by 5kg (10 jin, a traditional unit of weight) bait, take by weighing amylum adhesive 250g (5 liang), add warm water 1250ml (2.5 jin, a traditional unit of weight), paste is boiled in heating, cools to room temperature (must cool! ), take by weighing shrimp medicine 50g of the present invention (1 liang) again, add in the gelatinized corn starch, fully to mix thoroughly to rub with the hands lightly in the 5kg of falling people (10 jin, a traditional unit of weight) bait of back and mix evenly, airing can use.
Two kinds of compositions of Radix Phytolaccae powder and lysozyme also can prepare and meet into the shrimp medicine that the object of the invention requires, and are called " two-in-one " shrimp medicine, and at this moment the powder intermediate raw material of these two kinds of compositions feeds intake mixed by following proportioning:
Radix Phytolaccae powder: 40～90%
Lysozyme: 10～60%
Advantage of the present invention is:
(L) the various pathogenic bacterias of prawn are all had kill ability preferably, especially the antibacterial to prawn has remarkable effect.
(2) this product is a primary raw material with biochemical enzymes and plant mainly, environmentally safe, nontoxic, have no side effect, do not produce drug resistance.
(3) rely mainly on prevention, the control combination improves immunity, promotes the shrimp growth, enhance vigour, and be a kind of comprehensive shrimp medicine.
(4) good stability, it is convenient to preserve, and effect duration is long.
Below be embodiments of the invention:
Embodiment 1 is No. 1 prescription, Radix Phytolaccae powder 45%, lysozyme 8%, garlicin beta-schardinger dextrin-inclusion compound 4%, thorn boule 41%, Herb Gynostemmae Pentaphylli powder 2%.
Embodiment 2 is No. 4 prescriptions, wherein Radix Phytolaccae powder 48%, lysozyme 8%, garlicin beta-schardinger dextrin-inclusion compound 3.5%, thorn boule 38%, Herb Gynostemmae Pentaphylli powder 2.5%.
Embodiment 3 is No. 3 prescriptions, wherein Radix Phytolaccae powder 34%, lysozyme 7.2%, garlicin beta-schardinger dextrin-inclusion compound 3.5%, thorn boule 38%, Herb Gynostemmae Pentaphylli powder 2.5%
Embodiment 4 is No. 2 prescriptions, wherein Radix Phytolaccae powder 36%, lysozyme 7.5%, garlicin beta-schardinger dextrin-inclusion compound 4.3%, thorn boule 50%, Herb Gynostemmae Pentaphylli powder 2.2%
Embodiment 5 is No. 5 prescriptions: Radix Phytolaccae powder 49%, lysozyme 8.4%, garlicin beta-schardinger dextrin-inclusion compound 3.3%, thorn boule 37.7%, Herb Gynostemmae Pentaphylli powder 1.6%
Embodiment 6 is No. 6 prescriptions, wherein Radix Phytolaccae powder 46%, lysozyme 8.2%, garlicin beta-schardinger dextrin-inclusion compound 4%, thorn boule 40%, Herb Gynostemmae Pentaphylli powder 1.8%
Effect of the present invention:
Below be the laboratory results to No. 1, No. 2, No. 3, No. 4 four seed shrimp medicines are done:
1, material: available from prawn culturing field, prune town, Tongan City, Xiamen City, prawn is the long 3.9～7.1cm of body with shrimp in test, and the japonicus of body weight 0.6～3.1g has been determined band virus after testing. Trial drug adopts four kinds of pulvis No. 1 of the present invention, No. 2, No. 3, No. 4, and bait adopts common bait: the hippocampus board #2 synthetic diet A of university, B, medicinal bait: medicine of the present invention is admixed common bait by 1% amount, contains with egg white, dries for subsequent use.
2, method: the prawn of buying is supported temporarily in 26 cubic metres of Da Chi first, puts into respectively test tank after 3 days, 2 check experiment ponds common bait of only throwing something and feeding wherein, 4 test tanks pharmaceutical chemistry of throwing something and feeding No. 1, No. 2, No. 3, No. 4. Spread husky 2～3cm at the bottom of each pond, discharge water 1.5 tons. Drop into 56 tail prawns in each pond, every day, quantity of exchanged water 1/3, changes to suck residual bait at the bottom of the pond before the water, and a dead shrimp is removed, and recorded respectively water temperature variation every day, prawn death toll, prawn active situation, prawn body colour, appetite etc. after the bait throwing in.
3, result of the test:
Experimental session prawn Daily mortality amount and final survival rate statistical form
| 1＃ | 2# | 3＃ | 4＃ | Contrast A | Contrast B | |
| 17/11 | 56 | 56 | 56 | 56 | 56 | 56 |
| 18/11 | 3 | 2 | 2 | 0 | 5 | 2 |
| 19/11 | 5 | 7 | 5 | 5 | 5 | 6 |
| 20/11 | 3 | 8 | 5 | 5 | 6 | 5 |
| 21/11 | 5 | 8 | 2 | 1 | 7 | 9 |
| 22/11 | 4 | 3 | 5 | 2 | 5 | 5 |
| 23/11 | 5 | 2 | 4 | 2 | 4 | 4 |
| 24/11 | 1 | 2 | 2 | 2 | 4 | 0 |
| 25/11 | 0 | 0 | 1 | 1 | 0 | 1 |
| 26/11 | 1 | 2 | 2 | 1 | 2 | 4 |
| 27/11 | 1 | 0 | 0 | 0 | 0 | 1 |
| 28/11 | 0 | 0 | 1 | 1 | 2 | 1 |
| 29/11 | 1 | 0 | 1 | 0 | 0 | |
| 30/11 | 1 | 0 | 2 | 0 | 2 | 2 |
| 1/12 | 1 | 0 | 0 | 0 | 2 | |
| 2/12 | 0 | 0 | 1 | 1 | 0 | 1 |
| 3/12 | 1 | 0 | 1 | 0 | 1 |
| 1# | 2# | 3# | 4# | Contrast A | Contrast B | |
| 4/12 | 0 | 0 | 0 | 1 | 0 | 1 |
| 5/12 | 0 | 0 | 0 | 0 | 0 | 0 |
| 6/12 | 1 | 1 | 1 | 1 | 1 | 1 |
| Survive quantity | 18 | 7 | 6 | 31 | 10 | 4 |
| Survival rate | 39％ | 18％ | 14％ | 67％ | 18％ | 7.1％ |
Result of the test shows that No. 4 medicines are effective biologic products of preventing and treating prawn ' s virus, and the prawn survival rate reaches 67 %, exceeds the under equal conditions contrast pond 49%～59.9% of test, has the potentiality of applying.
Below be the Experiment on Function to the rate of body weight gain to Penaeus monodon that No. 4, No. 5, No. 6 three seed shrimp medicines are done, immunity, premunition, antiviral, antibacterium and survival rate aspect, existing that test situation and report the test is as follows:
One, test material
1, for the reagent thing: by Ningbo three kinds of biologic products providing of this thing biochemical product Co., Ltd, Shanghai hero research institute not: No. 4, No. 5 and No. 6. Wherein No. 4, No. 5 medicines are for slightly being yellow granular solids, and No. 6 medicines are greeny granular solids.
2, for the examination prawn: be the Penaeus monodon that the pond is propagated artificially, body weight is 4～6g, and body length is 6～8cm, and is all healthy anosis.
3, for examination virus: be white spot syndrome baculovirus (annotate: this virus is the cause of disease of national Explosive Epidemic Disease of Cultured Shrimp, but classification position and name are not yet unified).
4, for the examination bacterial classification: for Penaeus monodon causes bacterium vibrio alginolyticus (Vibrio alginolyticus) and Vibrio vulnificus (V.vulnificus).
5, for the examination feed:
1) normal diet: be commercially available eel powder-material, with the front water that suitably adds, with using after the hand-operated meat mincer extrusion process moulding.
2) pharmaceutical feed: with No. 4, No. 5, No. 6 biologic products are pressed and are used after the normal diet processing method is processed respectively by after 2% or 3% the amount elder generation and the abundant mixing of eel powder-material.
Provide by Chinese Academy of Science Nanhai Ocean Research Institute marine organisms virus research center for examination prawn, virus, bacterial classification and feed.
Two, test method
The Penaeus monodon of experiment usefulness is normally raised in outdoor large cement culturing pool (about 200 square meters of area, dark 1.5 meters of pond). Catch out during experiment, put at random in the indoor miniature water mud sump (area is 1 square meter, dark 1 meter of pond, inwall tiling, 0.6 ton in practical water body) by some and test.
(1) rate of body weight gain, survival rate, immunity and premunition test
1, rate of body weight gain and survival rate test
By the healthy Penaeus monodon that catches out the long 6～8cm of body in the outdoor large cement pit, random packet, every group of 35 tails, putting in the indoor miniature water mud sump of people hunger after weighing raises and train and begins test after 2 days, after raising 15 days under the continuous charge condition, weigh, and record the dead quantity of the accumulation of respectively organizing in 15 days, to measure biologic product to the impact (table 1) of Penaeus monodon rate of body weight gain and survival rate.
2, immunity determination test
After rate of body weight gain and the survival rate off-test, get prawn immediately and measure its immunity, get shrimp 5 tails for every group, measure its blood lymphocyte by tail and engulf luminous power.Adopt chemiluminescence (CL) method to measure, zymosan with the autoserum conditioning is an activator, luminol is the cold light agent, measures (table 2) with the SHG-1 type biochemiluminescence instrument (with 486 microcomputer couplings) that Experiment Plant of Shanghai Technical Supervision Bureau produces.
3, premunition test
After experiment 1 and 2 finishes, carry out immediately, get prawn 20 tails for every group, penetrate vibrio alginolyticus by endnote, every endnote is penetrated the bacterium liquid 0.1ml that concentration is 4 * 10/ml, observe the 96 hours dead quantity with interior prawn, duration of test is not thrown any feedstuff, to measure its premunition (table 3).
(2) antivirus test
By the healthy Penaeus monodon of catching out the long 6～8cm of body in the outdoor large cement pit, random packet, every group 35 tail, put into after indoor small-sized cement pond hunger raises and train 2 days, every group of sick shrimp (frozen in 20 ℃ of refrigerators) of Penaeus monodon of throwing something and feeding simultaneously and being infected by leukasmus rhabdovirus, throw something and feed by the amount of the sick shrimp of every tail 2.0 grams and to carry out the artificial challenge, infect back the 2nd day throw something and feed normal diet (matched group) and pharmaceutical feed (experimental group), observe incidence and the mortality rate of prawn in 7 days later on continuously, with the antivirus action (table 4) of detection of biological preparation.
(3) antibacterium test
By the healthy Penaeus monodon of catching out the long 6～8cm of body in the outdoor large cement pit, random packet, every group 35 tail put into after indoor small-sized cement pond hunger raises and train 2 days, penetrates by endnote that to contain the vibrio alginolyticus amount be 4.2 * 10
6/ ml or to contain trauma by other people vibrio amount be 2.1 * 10
6The bacterium liquid 0.1ml of/ml, and raise with normal diet (matched group) and pharmaceutical feed (experimental group), death toll and the survival rate of prawn in 7 days observed, with the antibacterium effect (table 5) of detection of biological preparation.
Three, result of the test
1, biological preparation is to the effect of Penaeus monodon weightening finish and survival rate
No. 4, biological preparation, what No. 5 and No. 6 and normal diet were raised healthy Penaeus monodon the results are shown in Table 1, as shown in Table 1, the survival rate of three kinds of drug test groups and matched group is all more than 82.8%, average every tail of Penaeus monodon increases day by day weight more than 0.1g, is not having marked difference (table 1) aspect survival rate and the rate of body weight gain between matched group and the drug test group.In addition, at No. 4, No. 5, No. 6 biological preparation add in the feedstuff with 2% and 3% amount respectively throws something and feeds, and the influence of healthy Penaeus monodon survival rate and rate of body weight gain is not had significant difference.
Table 1 biological preparation is to the influence of Penaeus monodon hypertrophy rate and survival rate
| Group | Shrimp number (tail) | Body weight (g) | Weightening finish (g) | Average daily gain (g/ tail) | Death toll (tail) | Survival rate (%) |
| No. 4 (2%) | 35 | 268 | 52.2 | 0.12 | 6 | 82.8 |
| No. 4 (3%) | 35 | 272 | 52.2 | 0.13 | 5 | 85.7 |
| No. 5 (2%) | 35 | 275 | 58.6 | 0.13 | 5 | 85.7 |
| No. 5 (3%) | 35 | 269 | 58.5 | 0.12 | 4 | 88.5 |
| No. 6 (2%) | 35 | 280 | 65.3 | 0.14 | 4 | 88.5 |
| No. 6 (3%) | 35 | 265 | 58.0 | 0.13 | 5 | 85.7 |
| Contrast | 35 | 270 | 58.1 | 0.13 | 5 | 85.7 |
2, biological preparation is to the influence of Penaeus monodon immunity
Three kinds of biological preparation see Table 2 to the testing result of Penaeus monodon hemolymph phagocyte chemiluminescence (CL), by table as seen: No. 5, No. 6 chemiluminescent background valuess of biological preparation processed group Penaeus monodon hemolymph phagocyte, all raise to some extent than matched group when the peak value of induced luminescence and peak or prolong, No. 4 the biological preparation processed group does not then have significant difference (table 2) with matched group.Table 2 biological preparation is to the influence of Penaeus monodon hemolymph chemiluminescence (CL) (X ± SD)
| Group | Shrimp number (tail) | Background (cpm/ml blood) | Peak value (cpm/ml blood) | During the peak (branch) |
| No. 4 (2%) | 5 | 76±28 | 1237±123 | 5.2±1.3 |
| No. 4 (3%) | 5 | 82±36 | 984±212 | 5.4±1.1 |
| No. 5 (2%) | 5 | 85±37 | 1388±231 | 6.3±1.2 |
| No. 5 (3%) | 5 | 96±33 | 1428±193 | 6.7±1.5 |
| No. 6 (2%) | 5 | 98±43 | 1376±213 | 6.8±1.4 |
| No. 6 (3%) | 5 | 103±57 | 1238±212 | 6.6±1.4 |
| Contrast | 5 | 78±24 | 875±141 | 5.2±1.7 |
3, biological preparation is to the effect of Penaeus monodon premunition
After raising with biological preparation continuously through 15 days, Penaeus monodon demonstrates certain resistance (table 3) to artificial attack vibrio alginolyticus, and wherein best with No. 6 biological preparation treatment effects, No. 5 biological preparation takes second place, No. 4 biological preparation is the poorest, but all slightly improves (table 3) than matched group.
Table 3 biological preparation is to the influence of Penaeus monodon premunition
| Group | Shrimp number (tail) | Attack cause of disease | Death toll (tail) | Survival rate |
| No. 4 (2%) | 20 | Vibrio alginolyticus | 19 | 5 |
| No. 4 (3%) | 20 | Vibrio alginolyticus | 19 | 5 |
| No. 5 (2%) | 20 | Vibrio alginolyticus | 16 | 20 |
| No. 5 (3%) | 20 | Vibrio alginolyticus | 14 | 30 |
| No. 6 (2%) | 20 | Vibrio alginolyticus | 13 | 35 |
| No. 6 (3%) | 20 | Vibrio alginolyticus | 12 | 40 |
| Contrast | 20 | Vibrio alginolyticus | 20 | 0 |
4, biological preparation is to the antiviral effect of Penaeus monodon
Penaeus monodon is after leukasmus rhabdovirus infects, promptly began to show manifest symptom on the 3rd day, mainly show as carapace and white macula occurs, carapace easily separates with the shrimp body rubescent with subcutaneous tissue, and beginning is dead successively, one all death of week all infected shrimps of internal reference group, but through No. 4, No. 5, after No. 6 biological preparation is handled, still have the part survival in one week, and the survival prawn there is not above-mentioned symptom appearance.In three kinds of biological preparation, best with No. 5 antiviral effects, 6 numbers it, No. 4 (table 4) the poorest.
Table 4 biological preparation is to the antiviral effect of Penaeus monodon
| Group | Shrimp number (tail) | Attack cause of disease | Death toll (tail) | Survival rate |
| No. 4 (2%) | 35 | Leukasmus rhabdovirus | 34 | 2.8 |
| No. 4 (3%) | 35 | Leukasmus rhabdovirus | 33 | 5.7 |
| No. 5 (2%) | 35 | Leukasmus rhabdovirus | 24 | 31.4 |
| No. 5 (3%) | 35 | Leukasmus rhabdovirus | 25 | 28.5 |
| No. 6 (2%) | 35 | Leukasmus rhabdovirus | 32 | 8.5 |
| No. 6 (3%) | 35 | Leukasmus rhabdovirus | 31 | 11.4 |
| Contrast | 35 | Leukasmus rhabdovirus | 34 | 2.8 |
5, biological preparation is to the effect of Penaeus monodon antibacterial
After the Penaeus monodon manual injection infected vibrio alginolyticus and wound vibrio, matched group was all dead in 48 hours, and with after the biological preparation processing survival being arranged all, what wherein survival rate was the highest is No. 6 biological preparation processed group, next is No. 5, the poorest No. 4, but all be higher than matched group (table 5).
Table 5 biological preparation is to the effect of Penaeus monodon antibacterial
| Group | Shrimp number (tail) | Attack strain | Death toll (tail) | Survival rate |
| No. 4 (2%) | 35 | Vibrio alginolyticus | 25 | 28.6 |
| No. 5 (2%) | 35 | Vibrio alginolyticus | 19 | 45.7 |
| No. 6 (2%) | 35 | Vibrio alginolyticus | 5 | 85.7 |
| No. 4 (2%) | 35 | Vibrio vulnificus | 23 | 34.3 |
| No. 5 (2%) | 35 | Vibrio vulnificus | 17 | 51.4 |
| No. 6 (2%) | 35 | Vibrio vulnificus | 6 | 82.8 |
| Contrast | 18 | Vibrio alginolyticus | 18 | 0 |
| 18 | Vibrio vulnificus | 18 | 0 |
Four, discuss
Above-mentioned result of the test shows, No. 4, " Fu Site " biological preparation all has enhancing Penaeus monodon premunition, immunity and antiviral, antibacterial effect in various degree No. 5 and No. 6.
No. 6 biological preparation has significant antibacterial action, under two kinds of morbid vibrios of Penaeus monodon (vibrio alginolyticus and Vibrio vulnificus) are attacked, still have survival rate up to 82.8～85.7%, the proof said preparation has significant curative effect to the bacterial disease of prawn, and said preparation also has effect preferably to improving prawn immunity and premunition simultaneously.Therefore, No. 6 preparation is a kind of antibiotic shrimp medicine with big potentiality, and shortcoming is that the anti-virus ability of said preparation is not strong.
No. 5 preparation has antiviral effect preferably, but can only improve 28～31% survival rate than matched group, it also has certain antibacterial action, because this test is to carry out under the laboratory experiment condition of continuous charge, used sea water also is that the superiority of its environmental condition is to be beyond one's reach under the nature cultivating condition, therefore through filtering sea water, whether said preparation has this good effect in actual applications, the further check of still needing.
Though the effect that No. 4 agent also have certain antiviral, antibacterium and improve premunition all not as No. 5 and No. 6 preparation ideals, must further improve, just might be as the medicinal application of a kind of prevention and cure of shrimp disease preferably.
Five, brief summary
1, No. 4, " Fu Site " biological preparation, No. 5, No. 6 equal tools improve Penaeus monodon immunity and premunition effect, has the effect of antiviral and antibacterium, wherein best with No. 6 preparation antibacterial actions, No. 5 preparation has antivirus action preferably, No. 4 preparations at antibiotic and anti-virus aspect all not as No. 6 and No. 5 preparations.
2,, show No. 6 and No. 5 biological preparation are respectively effective preparations of preventing and treating prawn bacterial disease and virosis to have application value by to the mensuration of Penaeus monodon immunity, premunition, antiviral, antibacterium and survival rate.
Claims (9)
1, a kind of new drug of prawn disease, it includes two kinds of necessary compositions, and a kind of is the Radix Phytolaccae powder, and another is a lysozyme.
2, the new drug of prawn disease as claimed in claim 1 is characterized in that: described Radix Phytolaccae powder can replace with the various pokeweed antiviral proteins that utilization protein separation method obtains.
3, the new drug of prawn disease as claimed in claim 1 is characterized in that: described composition includes Radix Phytolaccae powder, lysozyme, garlicin beta-schardinger dextrin-inclusion compound, thorn boule and Herb Gynostemmae Pentaphylli powder.
4, the new drug of prawn disease as claimed in claim 3 is characterized in that: the ratio of various compositions (percentage by weight) is in the described compatibility:
Radix Phytolaccae powder 34.71～50.14%
Lysozyme 7.12～8.70%
Garlicin β-
Cyclodextrin 3.03～4.38%
Inclusion compound
Thorn boule 37.61～53.04%
Herb Gynostemmae Pentaphylli powder 1.54～2.32%
5, a kind of manufacture method of prawn disease new drug, it includes makes the powder intermediate raw material earlier with the compatibility of five kinds of compositions, and then by preparing patent medicine as the formula proportion of claim 4 defined, described five kinds of compositions are: Radix Phytolaccae powder, lysozyme, garlicin beta-schardinger dextrin-inclusion compound, thorn boule, Herb Gynostemmae Pentaphylli powder.
6, the manufacture method of prawn disease new drug as claimed in claim 5 is characterized in that: the preparation of described Radix Phytolaccae powder is, at first bright Radix Phytolaccae cleaned, cutting, drying, pulverizing, sieves at last, and granularity is controlled between 50 order to 80 orders.
7, the manufacture method of prawn disease new drug as claimed in claim 5, it is characterized in that: the preparation technology of described garlicin beta-schardinger dextrin-inclusion compound intermediate raw material is: get in the water that beta-schardinger dextrin-is dissolved in 12 times of weight, temperature is controlled at 20～30 ℃, drip and amplify Bulbus Allii oil, constantly stirred five hours, standing over night is extremely done with the vacuum pump sucking filtration, dry being roughly under 50 ℃ the temperature, be crushed to and be roughly 80 purpose granularities.
8, the manufacture method of prawn disease new drug as claimed in claim 5, it is characterized in that: the preparation technology of described thorn boule is: the system of the measuring pears natural juice by weight 40%, the medical starch of adding 60%, put into mixer and make soft material, granulate with 24 purpose screen clothes, dry below 50 ℃, get 24 purpose powder, be crushed to again and be roughly 80 purpose granularities.
9, the new drug of prawn disease as claimed in claim 1 is characterized in that: the mixed proportion of described Radix Phytolaccae powder and lysozyme is:
Radix Phytolaccae powder: 40～90%
Lysozyme: 10～60%
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97106607A CN1067887C (en) | 1997-09-15 | 1997-09-15 | New drug for preventing and curing prawn disease and its manufacturing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN97106607A CN1067887C (en) | 1997-09-15 | 1997-09-15 | New drug for preventing and curing prawn disease and its manufacturing method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1211425A CN1211425A (en) | 1999-03-24 |
| CN1067887C true CN1067887C (en) | 2001-07-04 |
Family
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| Application Number | Title | Priority Date | Filing Date |
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| CN97106607A Expired - Fee Related CN1067887C (en) | 1997-09-15 | 1997-09-15 | New drug for preventing and curing prawn disease and its manufacturing method |
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| CN (1) | CN1067887C (en) |
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| CN102671190A (en) * | 2012-04-26 | 2012-09-19 | 安徽省芬格欣普蓝生物药业有限公司 | Enzymic preparations for improving human body immunity and enhancing gastrointestinal and respiratory functions |
| CN106983771B (en) * | 2017-05-27 | 2020-01-31 | 海南热带海洋学院 | Medicine and composition for preventing and treating hippocampus head ulceration |
| CN109452497A (en) * | 2018-12-29 | 2019-03-12 | 盐城工学院 | A kind of additive and the preparation method and application thereof preventing South America prawn white excrement disease |
| CN110343656A (en) * | 2019-08-15 | 2019-10-18 | 中国水产科学研究院淡水渔业研究中心 | A kind of separation method of Procambius clarkii blood lymphocyte |
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1997
- 1997-09-15 CN CN97106607A patent/CN1067887C/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| 中成药研究，1986,(12),24-25 1986.1.1 韩克惠，商陆的免疫药理和临床应用 * |
| 同济医科大学学报，1991,20(6),365-368 1991.1.1 褚嘉佑等，三种单抗偶联PAP-S制备的免毒素对人白摁病T淋巴细胞的细胞毒性作用 * |
| 同济医科大学学报，1991,20(6),365-368 1991.1.1 褚嘉佑等，三种单抗偶联PAP-S制备的免毒素对人白摁病T淋巴细胞的细胞毒性作用;中成药研究，1986,(12),24-25 1986.1.1 韩克惠，商陆的免疫药理和临床应用 * |
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