CN103583570A - Multi-effect pesticide for broad-spectrum sterilization and virus resistance in sericultural production process and application of pesticide - Google Patents
Multi-effect pesticide for broad-spectrum sterilization and virus resistance in sericultural production process and application of pesticide Download PDFInfo
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- CN103583570A CN103583570A CN201310579002.6A CN201310579002A CN103583570A CN 103583570 A CN103583570 A CN 103583570A CN 201310579002 A CN201310579002 A CN 201310579002A CN 103583570 A CN103583570 A CN 103583570A
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Abstract
The invention belongs to the technical field of sericultural production and in particular relates to a multi-effect pesticide for broad-spectrum sterilization and virus resistance in a sericultural production process and an application of the pesticide. The pesticide is 20-100 ppm of a nanosilver solution. A test on a lab plate discovers that after 20-100 ppm of the nanosilver solution is used to treat major pathogens of silkworm for 8 hours, the nanosilver solution has a strong killing effect on various pathogens in the sericultural production process and is capable of thoroughly disinfecting cocooneries, utensils and sericultural surrounding environments in one time; a use method is simple and quick and is easily accepted by masses of silkworm farmers; the purposes of achieving a disinfecting effect, preventing diseases and increasing the economic benefit of the sericultural production are achieved.
Description
Technical field
The invention belongs to silkworm production technical field, be specifically related to a kind of antiviral multiple effective medicine of broad-spectrum sterilization and application thereof on silkworm produces.
Background technology
At present, the disinfecting drug that silkworm and mulberry are used in producing, all exists fungicidal spectrum inadequate, and cannot kill virus disease, the problem of the Different Kinds of Pathogens such as fungi.Such as present widely used bleaching powder, its active ingredient is available chlorine, deposit a period of time after active ingredient easily volatilize, and fungicidal spectrum is more limited, and virus disease and other pathogens are significantly declined; And sterilization time has strong corrosion effect to cotton goods, ironware etc.On silkworm and mulberry are produced, general disinfecting drug can not reach the effect of thorough disinfection, thereby is difficult to carry out the Eight Terms Guidelines of " putting prevention first, integrated control ".Silkworm main pathogen polyhedrosis virus, soldier's gemma of falling bacterium, black chest sepsis bacterium gemma, white deadlock, green deadlock, aspergillus spore etc. have strong killing action
Nano Silver (Nanoparticle Silver) is exactly that particle diameter is accomplished to nano level argent simple substance.Nano Silver particle diameter mostly, in 25 nanometer left and right, has strong inhibition and killing action to tens of kinds of pathogenic microorganismss such as Escherichia coli, gonococcus, chlamydia trachomatises, and can not produce drug resistance.But Nano Silver as whether polyhedrosis virus, soldier's gemma of falling bacterium, black chest sepsis bacterium gemma, white deadlock, green deadlock, aspergillus spore and particulate protozoon etc. have killing action, does not have report for silkworm main pathogen so far.
Summary of the invention
The technical problem to be solved in the present invention is in order to overcome silkworm and mulberry, to produce the deficiency of existing disinfecting drug, provide a kind of on silkworm produces the antiviral multiple effective medicine of broad-spectrum sterilization, this medicine fungicidal spectrum is wide, silkworm virus disease, fungi are all had to strong killing action.
Object of the present invention is achieved by the following technical programs:
Silkworm and mulberry are produced the antiviral multiple effective medicine of broad-spectrum sterilization, the Nano silver solution that described medicine is 20～100ppm.
The preparation method of the Nano silver solution of described 20～100ppm is: with particle diameter be less than 5nm, valid density is that the commercially available Nano Silver product of 1000ppm is as mother liquor, in water: the ratio that mother liquor is 9～49:1 adds water in mother liquor, after diluting the Nano silver solution of 20～100ppm.
The present invention by experiment chamber plate test finds, with the Nano silver solution of 20～100ppm, processes silkworm main pathogen 8 hours, and the various pathogens that Nano silver solution is produced silkworm and mulberry have strong killing action.Therefore, the invention provides a kind of Nano silver solution, can be widely used in killing silkworm main diseases original medicine.Described silkworm pathogen is that black chest sepsis bacterium, soldier fall bacillus, gold-coloured staphylococci, mulberry tree red rust bacterium, silkworm blood type pus illness (NPV) or/and bombyx mori cell cytoplasmic polyhedrosis virus (CPV).
The invention has the beneficial effects as follows:
The present invention finds first, use the Nano silver solution of 20～100ppm to there is strong killing action to the various pathogens of silkworm and mulberry production, can disposablely to cocoonery, apparatus, sericulture surrounding environment, carry out thorough disinfection, using method simple and fast, be easily that numerous silkworm raisers accept, thereby reach sterilizing prevent disease, improve the object of silkworm and mulberry production economy benefit.
figure of description
Fig. 1. the Nano silver solution of variable concentrations is measured the fungicidal spectrum of Nano Silver to the fungicidal spectrum Plating of the various silkworm phytosis originals; I is the bactericidal action of variable concentrations Nano Silver to septicaemia, and II is that variable concentrations Nano Silver is to the bactericidal action of bacillus of soldier; III is the bactericidal action of variable concentrations Nano Silver to gold-coloured staphylococci; IV is the bactericidal action of variable concentrations Nano Silver to mulberry tree red rust bacterium.
Fig. 2. the bactericidal effect of Nano silver solution to septicaemia; I is the septicaemia bacterium photo that Nano Silver is processed, and in figure I, A thalline contains a large amount of nano-Ag particles (being shown as white), and B is that Nano Silver kills the bacterium remains after thalline; II is the contrast that phosphate buffer is processed.
Fig. 3. the action effect of Nano silver solution to silkworm blood type pus illness (NPV); I is the NPV photo that Nano Silver is processed; II is the contrast that phosphate buffer is processed.
Fig. 4. the action effect of Nano silver solution to bombyx mori cell cytoplasmic polyhedrosis virus (CPV); I is the CPV photo that Nano Silver is processed; II is the contrast that phosphate buffer is processed.
Fig. 5 .NPV adds food silkworm laboratory rearing situation.
Fig. 6. the biological effect that Nano silver solution processing blood type pus illness is added food silkworm detects; Wherein scheme A and confirmed that the NPV pathogenicity after Nano silver solution is processed declines greatly, and the NPV pathogenicity that figure B control group proof is processed without Nano silver solution is high; Figure C, D are blank, comprise Sumerian Silver and distilled water contrast.
Embodiment
Below in conjunction with Figure of description and specific embodiment, further describe the present invention.Unless stated otherwise, reagent, the equipment that the present invention adopts is the conventional reagent of the art and equipment.
The present invention's Nano Silver used is Ukraine commercially available Nano Silver product Sumerian Silver, and this product Nano Silver particle diameter is less than 5nm; Mother liquor valid density is 1000ppm.
1. configure the Nano silver solution of variable concentrations: the concentration of Nano silver solution is mainly 2.5ppm, 5ppm, 10ppm, 20ppm, 100ppm.
Collocation method is: with Ukraine commercially available Nano Silver product Sumerian Silver, as mother liquor, (this product cut size is less than 5nm, mother liquor valid density is the colourless transparent solution of 1000ppm), respectively in mother liquor: the ratio of water=1:399,1:199,1:99,1:49,1:9, in mother liquor, add rural well water or running water, directly after dilution, obtain the Nano silver solution of 2.5ppm, 5ppm, 10ppm, 20ppm, 100ppm.
2. measure the antimicrobial spectrum of above-mentioned Nano silver solution to silkworm main pathogen: with silkworm main pathogen as black chest sepsis bacterium, soldier fall bacillus (
bacillus thuringinsis), gold-coloured staphylococci (
streptococcus aureus), mulberry tree red rust bacterium (
aecidium mori Barclay), silkworm blood type pus illness (NPV), bombyx mori cell cytoplasmic polyhedrosis virus (CPV) is as determination object, concrete assay method is:
Bacterium and fungi sample: various pathogens are inoculated into respectively to LB liquid nutrient medium, at 28～37 ℃, cultivate 8～12 hours (fungi sample is put 28 ℃).Counting.With 1 х 10
8bacterium/mL is as standard bacterium liquid.On super-clean bench, to the solid culture medium that adds 5mL to dissolve in standard culture (diameter 12mm), when its temperature drops to 40 ℃, add 200 μ L standard bacterium liquid, do not solidify and frontly mix fast.Solidify and with card punch, punch afterwards.Then, Xiang Kongzhong adds respectively the Nano silver solution of variable concentrations, and sealing is inverted in 4 ℃ of refrigerators, spreads 1 hour.All be placed in afterwards 28～37 ℃ of incubators and cultivate 4～8 hours, observe, take pictures.
Virus N PV and CPV sample: viral polyhedron concentration is 2 х 10
8cFU/mL, gets 1mL sample and uses 100ppm Nano Silver room temperature treatment 8 hours, according to electron microscopic sample preparation method, prepares electron microscopic observation sample.
3. measurement result: 20～100ppm Nano Silver is acted on to bacterial disease of domestic silkworm 8 hours, gram-positive bacteria and negative bacterium are had to stronger bactericidal effect, the results are shown in Figure 1,2,3,4.In Fig. 1, wherein I is the bactericidal action of variable concentrations Nano Silver to septicaemia, and the Nano silver solution of 2.5ppm demonstrates good bactericidal effect; II be variable concentrations Nano Silver to soldier fall bacillus (
bacillus thuringinsis) bactericidal action, valid density is between 2.5～5ppm; III be variable concentrations Nano Silver to gold-coloured staphylococci (
streptococcus aureus) bactericidal action, valid density is between 5～10ppm; IV be variable concentrations Nano Silver to mulberry tree red rust bacterium (
aecidium moribarclay) bactericidal action, valid density is between 20～100ppm.From Fig. 3, wherein I is the NPV photo that Nano Silver is processed; II is the contrast that phosphate buffer is processed, in the NPV viral inclusion body of the figure I in Fig. 3 baculovirus particles with compare more at randomly, near inclusion body edge, and the baculovirus particles of contrast concentrates on inclusion body middle section.From the above results, can find out: for example to septicaemia (
bacillus spp.), working concentration just can reach practical bactericidal effect at 2.5ppm; The Nano silver solution of 100ppm acts on silkworm virus disease 8 hours, and variation to a certain degree occurs the polyhedrin of virus (NPV and CPV), and particularly to CPV, profile variation is remarkable; Working concentration 20～100ppm can kill mulberry tree red rust (
aecidium mori Barclay), this is a kind of fungal disease.
Use the Nano silver solution of 20～100ppm to there is strong killing action to the various pathogens of silkworm and mulberry production, can disposablely to cocoonery, apparatus, sericulture surrounding environment, carry out thorough disinfection, using method simple and fast, be easily that numerous silkworm raisers accept, thereby reach sterilizing prevent disease, improve the object of silkworm and mulberry production economy benefit.
Embodiment 2
Nano silver solution described in embodiment 1, in laboratory small-scale feeding experiment, after research Nano silver solution processing blood type pus illness (NPV) and cytoplasmic polyhedrosis virus (CPV), adds food and raises the impact on silkworm morbidity.
The enforcement time: on March 23rd, 2013 was to April 22.
Implement place: Agricultural University Of South China in the school, insect Physiology and biochemistry laboratory, building, Asian-Pacific area silkworm and mulberry training center north.
Implementing method:
Cultivated silkworm breed variety: original seed 7532; Mulberry leaf: pick up from Agricultural University Of South China mulberry field, kind is Guangdong Jing Sang.
Freshen food method: with Nano Silver, process CPV and NPV respectively, together with CPV, NPV, test, and set up blank respectively.Wherein, Nano Silver working concentration is 2ppm.The original storage liquid concentration of CPV is 1 * 10
8cFU/mL, the original storage liquid concentration of NPV is 2 * 10
8cFU/mL, dilutes 100 times as working concentration.Processed group: join in NPV and each 100mL working concentration liquid of CPV room temperature treatment 8～12h according to Nano Silver working concentration.Each 100mL working concentration of control group: NPV and CPV, with processed group room temperature 8～12hr.Blank group: use equivalent sterile water.
Prepare after various treatment fluids, with spray bottle, be evenly sprayed onto mulberry leaf surface, after drying for 4 age silkworm within the 3rd day, freshen food.
Zoning design: carry out subregion in 4 ages during silkworm, NPV and CPV are tested respectively, experimental group is respectively that NPV, CPV, Nano Silver are processed NPV, Nano Silver is processed CPV, and three repetitions are established in each experiment, and each repeats to set up blank (adding food equivalent sterile water).
Result of implementation 1: to 5 later stages in age, three experimental group that NPV freshens food find that there is a large amount of silkworm Mortalities, and the NPV experimental group incidence of disease of processing through Nano Silver roughly with blank group quite (table 1, Fig. 6).
Table 1 Nano Silver is processed NPV experiment statistics table
Result of implementation 2: the CPV virulence of processing through Nano Silver significantly declines, its premunition even surpasses blank group (table 2).During matured silkworm is placed on small straw bundles to spin cocoons, it is more that untreated CPV lands silkworm, migration silkworm, sick silkworm quantity; After Nano Silver is processed, CPV virulence obviously declines.
Table 2 Nano Silver is processed CPV experiment statistics table
Remarks: between implementation period, rainwater is more, it is how wet that feeding environment belongs to normal temperature.The pathogenetic kind of silkworm is mainly NPV, CPV virus disease and bacteriosis coinfection.Check plot morbidity is mainly bacteriosis and stiff sick.
Data statistics
tcheck:
(1) when df=4, look into critical
tvalue,
t 0.005 (4)=5.598, |
t| >t
0.005 (4), negate H
0:
1=
2, p<0.005, shows that the NPV processing with Nano Silver and the NPV not processing with Nano Silver are very remarkable to the pathogenic difference on effect of silkworm.
(2) when df=4, look into critical
tvalue,
t 0.005 (4)=5.598, |
t| >t
0.005 (4), negate H
0:
1=
2, p<0.005, shows that the CPV processing with Nano Silver and the CPV not processing with Nano Silver are very remarkable to the pathogenic difference on effect of silkworm.
Embodiment 3
Nano silver solution described in embodiment 1 is specifically applied in silkworm and produces.
Enforcement time, place: within 2013, breed silkworms season respectively three of spring, summer, autumn, choose at random 6 silkworm raisers test that carries out disinfection in Na Wu town, Huazhou City, Hejiang town, the whole year, totally 18 families participated in trying out.Wherein, each in seasons 3 people's nano-silver antiseptic solution, 3 people contrast with bleaching powder disinfectant.Cultivated silkworm breed variety is 9 cottonrose hibiscus X7 Hunan.
Sterilization method: man of preparation 100ppm Nano silver solution ，Yong peasant household well water is pressed water: dilution proportion mother liquor=9:1(v/v), is fully uniformly mixed.With sterilization, with spray gun, disinfection of silkworm rearing room is educated in grown silkworm ground, require spraying even, till moistening metope, ground.Other sericulture facilities are as the same in sterilization methods such as silkworm-storing chamber, corridor, silkworm room pad bricks.The sterilization of bleaching powder is prepared according to a conventional method, and application process is the same.
Result of implementation: rural area silk cocoon is produced the effect (as table 3～6, the date is for selling cocoon day) of nano-silver antiseptic solution on probation:
Breed silkworms and try out the Disinfection Effect 2013-6-13 of Nano Silver in table 3, rural area, Huazhou City
Breed silkworms and try out the Disinfection Effect 2013-9-9 of Nano Silver in table 4, rural area, Huazhou City
Breed silkworms and try out the Disinfection Effect 2013-10-30 of Nano Silver in table 5, rural area, Huazhou City
Table 6 Nano Silver rural area data statistic analysis on probation in 2013
From table, data are not difficult to find out, Nano Silver on probation is as disinfecting drug and traditional disinfecting drug bleaching powder comparison, and its individual to produce cocoon more lower slightly than bleaching powder with individual output value.Disinfecting drug is more importantly from the judgement of cocoon quality aspect, because morbidity in 5 latter stages in age most likely produces dead worm cocoon, directly to affect cocoon quality.Its average cocoon valency is compared with bleaching powder,
tassay difference is not remarkable.
Nano-silver antiseptic solution and bleaching powder disinfectant comparison: in the situation that average cocoon valency is suitable, that the use advantage of Nano Silver is is nontoxic, odorless, non-corrosiveness, particularly in Rural Furniture during many ironys article, long-time use can not corroded completely.But current cost ratio bleaching powder is slightly expensive.The advantage of bleaching powder is that Jiu Wei peasant household is used, cheap, but smell is pungent, corrosivity is extremely strong, and rearing instrument, cocoonery mostly are bleaching powder corrosion, and make cocoonery, doorframe etc. variegated for a long time, affect new rural area attractive in appearance.
Claims (4)
1. Nano silver solution is killed the application in silkworm main diseases original medicine in preparation.
2. application according to claim 1, is characterized in that, described silkworm pathogen is that black chest sepsis bacterium, soldier fall bacillus, gold-coloured staphylococci, mulberry tree red rust bacterium, silkworm blood type pus illness or/and bombyx mori cell cytoplasmic polyhedrosis virus.
3. the antiviral multiple effective medicine of broad-spectrum sterilization on silkworm and mulberry are produced, is characterized in that the Nano silver solution that described medicine is 20～100ppm.
4. medicine according to claim 3, it is characterized in that, the preparation method of the Nano silver solution of described 20～100ppm is: with particle diameter be less than 5nm, valid density is that the commercially available Nano Silver product of 1000ppm is as mother liquor, in water: the ratio that mother liquor is 9～49:1 adds water in mother liquor, after diluting the Nano silver solution of 20～100ppm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103947614A (en) * | 2014-04-24 | 2014-07-30 | 苏州大学 | Method for inhibiting in-vivo BmNPV viral multiplication of silkworms |
| CN106376530A (en) * | 2016-08-30 | 2017-02-08 | 盐城市大丰区蚕桑技术指导站 | Silkworm breeding method for preventing silkworm diseases |
| CN111588735A (en) * | 2020-05-29 | 2020-08-28 | 西南大学 | Application of nano silver in preparation of silkworm microparticle medicament for inhibiting main pathogen of silkworms |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103947614A (en) * | 2014-04-24 | 2014-07-30 | 苏州大学 | Method for inhibiting in-vivo BmNPV viral multiplication of silkworms |
| CN106376530A (en) * | 2016-08-30 | 2017-02-08 | 盐城市大丰区蚕桑技术指导站 | Silkworm breeding method for preventing silkworm diseases |
| CN111588735A (en) * | 2020-05-29 | 2020-08-28 | 西南大学 | Application of nano silver in preparation of silkworm microparticle medicament for inhibiting main pathogen of silkworms |
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