CN104784194A - Applications of natamycin in killing of ichthyophthirius multifiliis and cryptocaryon irritans - Google Patents
Applications of natamycin in killing of ichthyophthirius multifiliis and cryptocaryon irritans Download PDFInfo
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- CN104784194A CN104784194A CN201510082499.XA CN201510082499A CN104784194A CN 104784194 A CN104784194 A CN 104784194A CN 201510082499 A CN201510082499 A CN 201510082499A CN 104784194 A CN104784194 A CN 104784194A
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- natamycin
- control agent
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- little melonworm
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Abstract
The invention belongs to the technical field of chemical product uses, and discloses a prevention and control agent for killing ichthyophthirius multifiliis and cryptocaryon irritans outside aquatic animal bodies, wherein the prevention and control agent comprises, by weight, 5-25% of natamycin, 5-10% of a surfactant, 10-20% of a dissolution agent, and the balance of a solvent. According to the present invention, the natamycin is used, such that advantages of low toxicity, no toxicity on aquatic animals, and less residue are provided; the specific dissolution agent and the specific solvent are utilized, such that the dissolving effect is good, and the dissolution agent can help the entering of the prevention and control agent into the body so as to provide the significant ichthyophthirius multifiliis killing effect; and the prevention and control agent has characteristics of simple preparation, easy use, and low production cost.
Description
Natamycin is being killed little melonworm and is being stimulated the application in cryptonucleus insect.
Technical field
The invention belongs to chemical products purposes technical field, be specially natamycin and killing little melonworm and stimulating the application in cryptonucleus insect.
Technical background
Natamycin (Natamycin) is a kind of natural antifungal compound produced by streptomycete fermentation, belong to polyene macrolides, both various mycete, saccharomycetic growth extensively can effectively be suppressed, again can the generation of Antifungi toxin, the fresh-keeping and antifungal therapy of food antiseptic can be widely used in.Its structural formula is:
Natamycin, molecular formula is C
33h
47nO
13, faint yellow or beige powder, fusing point is 280 DEG C, is slightly soluble in water, is insoluble in majority of organic solvent.It relies on the sterol compound effect on its lactonic ring structure and fungal cell membrane, forms antibiotic-sterol compound, thus destroys the structure of the cytoplasma membrane of fungus.The hydrophilic segment (many alcohol moieties) of macrocyclic lactone forms water hole on film, damaging cells membrane permeability, and then cause aminoacid in bacterium, the materials such as electrolyte ooze out, thalline is dead, be a kind of filamentous fungi inhibitor such as yeast and mycete of natural, wide spectrum, highly effective and safe, it can not only Antifungi, can also prevent the generation of mycotoxin.Natamycin is harmless, is difficult to be absorbed by human body alimentary canal, and microorganism is difficult to produce resistance to it, simultaneously because of features such as its dissolubility are very low, is generally used for the surface anticorrosion of food.And natamycin is not also reported parasitic killing action.
Little melonworm and stimulation cryptonucleus insect are the important parasites that almost can infect all fresh water and seawater fish, cause huge harm to aquatic products industry.And to treat these two kinds of the most effective medicines of parasite be Mercury protonitrate. and malachite green oxalate, and these two kinds of medicines all have carcinogenic effect of Denging, so disabled.Along with the forbidding of these two kinds of medicines, domestic and international at present formaldehyde, copper sulfate, potassium permanganate, bronopol etc. are mainly adopted to these two kinds of parasitic controls, but these medicines are to little melonworm and stimulate cryptonucleus insect not have significant curative effect, life-time service brings the problems such as drug residue, environmental pollution, drug resistance simultaneously, is thus badly in need of these two kinds of specific medicines of parasite.
Summary of the invention
An object of the present invention is the novelty teabag providing natamycin, is namely killing little melonworm and is stimulating the application in cryptonucleus insect.
Further saying, is preparing the external little melonworm of posting of killing aquatic animal and stimulating the application in cryptonucleus insect medicine.
Further say, one of main purpose of the present invention is to provide natamycin to kill the outer little melonworm of Fish in preparation and stimulate the application in cryptonucleus insect medicine.
Two of object of the present invention there is provided the external little melonworm of a kind of killing aquatic animal and stimulates the control agent of cryptonucleus insect, it comprises the raw material of following weight percents: natamycin 5% ~ 25%, surfactant 5% ~ 10%, lytic agent 10 ~ 20%, all the other are solvent, and the summation of above-mentioned raw materials is 100%.
As preferably, described surfactant is tween.
Further, described surfactant is tween 20 or tween 80.
As preferably, described lytic agent is dimethyl sulfoxide or DMF or glacial acetic acid.
As preferably, described solvent is water.
Three of object of the present invention is to provide the preparation method of above-mentioned control agent, carries out according to the following steps:
1) natamycin, surfactant, lytic agent and solvent is taken by described weight portion, for subsequent use;
2) lytic agent is added in natamycin, and be stirred to natamycin and dissolve completely;
3) surfactant and solvent are added step 2) stir in the mixed solution of gained, obtain control agent.
Control agent using method of the present invention is simple, directly presses 15-25 mL/m
3dosage splash in pond can effectively preventing or kill the outer little melonworm of Fish and stimulate cryptonucleus insect.
The present invention adopts natamycin, and toxicity is little, to aquatic animal nonhazardous, remains less; Utilize specific lytic agent and solvent, solute effect is good, and lytic agent can assist control agent to enter in little melonworm body, remarkable to the killing effect of little melonworm; The preparation of control agent is simple, easy to use, and production cost is low.
Accompanying drawing explanation
Accompanying drawing 1 is the killing effect of natamycin to little melonworm larva;
Accompanying drawing 2 is the killing effect of natamycin to cryptocaryon irritans larva.
Detailed description of the invention
For a more clear understanding of the present invention and the drug effect of medicament of the present invention, the present invention is described in further detail for the specific embodiment completed according to technical solution of the present invention provided below by way of inventor, these are only the good embodiments of the present invention, but are not limited to these embodiments.
Embodiment 1:
Take natamycin 5g, stirring and dissolving in 10g dimethyl sulfoxide, take surface active agent tween-20 5g, aqueous solvent 80g, namely mixing and stirring obtains control agent.
Embodiment 2:
Take natamycin 10g, stirring and dissolving in 20g glacial acetic acid, take surface active agent tween-20 10g, aqueous solvent 60g, namely mixing and stirring obtains control agent.
Embodiment 3:
Take natamycin 15g, stirring and dissolving in 20g DMF, take surface active agent tween-80 10g, aqueous solvent 55g, namely mixing and stirring obtains control agent.
Embodiment 4:
Take natamycin 25g, stirring and dissolving in 20g DMF, take surface active agent tween-80 10g, aqueous solvent 45g, namely mixing and stirring obtains control agent.
Above-described embodiment can be enumerated a lot, this does not enumerate again, the digital proof that the great many of experiments done from applicant obtains, as long as select according to the parameter area of technical scheme of the present invention, can prepare the little melonworm of the aquatic animal reaching the object of the invention and stimulate the control agent of cryptonucleus insect.
Above-mentioned control agent can directly be splashed in pond, and with the pond water yield for reference, using dosage is 15-25 mL/m
3.
Be below drug effect of the present invention respectively to little melonworm with stimulate the test of cryptonucleus insect:
one, the test of pesticide effectiveness of parasitic little melonworm is killed
1 materials and methods
1.1 test fishes
The Ctenopharyngodon idellus (67.8 ± 5.6g) of the little melonworm of severe infections is from Zhejiang Institute of Fresh Water Aquatic Products seed base.Random selecting 10 tail fish, get holobranch microscopy and observe little melonworm quantity on the statistics gill, the little melonworm infection rate of test fish is 100%, and every tail fish on average infects little melonworm about 110.
Test medication is the control agent according to embodiment 1.
1.2 external insecticidal tests
The Ctenopharyngodon idellus of little for severe infections melonworm is put into beaker, and after little melonworm is swum out of, with suction pipe, plate is put in little for maturation melonworm collection, a part of polypide is used for the cultivation of little melonworm larva, and a part is used for the collection of encapsulation.
1.2.1 to the external insecticidal test of little melonworm germling
Test is carried out on 24 porocyte culture plates, adds the control agent (5.0,10.0,15.0,20.0 and 25.0mg/L of about 2ml variable concentrations in each cell hole
-1) and about 100 little melonworm larvas, after medication, 15min, 1h, 2h, 3h, 4h microscopy is observed once, adds up the little melonworm mortality rate in each hole after 4h.Test arranges dimethyl sulphoxide control group and fully sudden and violent gas Natural Water matched group, and often group test in triplicate.
The dead criterion of little melonworm: polypide cilium does not move, and kytoplasm does not flow, and after birth breaks, nuclear fragmentation.
1.2.2 to the external insecticidal test of little melonworm encapsulation
Test is carried out on 24 porocyte culture plates, adds control agent (5.0,10.0,15.0,20.0 and 25.0 mg/L of about 2ml variable concentrations in each cell hole
-1) and 30 little melonworm encapsulations, after medication, 4h microscopy is observed once, adds up the little melonworm encapsulation mortality rate in each hole after 4h.Test arranges dimethyl sulphoxide control group and fully sudden and violent gas Natural Water matched group, and often group test in triplicate.
The dead criterion of little melonworm encapsulation: encapsulation does not divide or encapsulation does not hatch larva.
The test of little melonworm is killed in 1.3 bodies
Be 1m at a volume
-3cement pit in put into 120 tail health and do not infect any parasitic Ctenopharyngodon idellus, support after one week temporarily, put into 600000 little melonworm larvas and infect, after infecting 24h, have the Ctenopharyngodon idellus of little melonworm to divide into groups infection, often organize 10 tail fishes, and put into the fish jar of 100L.The insecticidal test concentration arranging control agent according to external insecticidal test is 15.0,18.0 and 20.0 mL/m
3.The Natural Water into abundant aeration is contained, pH 7.0 ~ 7.5, water temperature 25 ± 1 DEG C in the glass jar that volume is 80cm × 60cm × 40cm.In water body, add the control agent of variable concentrations according to the concentration set, and stir, and at the 3rd day and the 5th day, each test group added the control agent renewed again.
When testing the 5th day, each concentration group takes out 3 tail Ctenopharyngodon idellus, and puts into beaker, collects encapsulation, and observe death and the division situation of encapsulation according to the method for external insecticidal test.
Test the 10th day, by all groups Ctenopharyngodon idellus take out, holobranch film-making, microscopy, add up every tail Ctenopharyngodon idellus in gill portion little melonworm amount of survival, calculate average killing rate.
Little melonworm Average Survival quantity × 100% of killing rate (%)=(the little melonworm Average Survival quantity of the little melonworm Average Survival quantity-test group of matched group)/matched group
Fish quantity × 100% of fish average mortality (%)=(fish amount of survival during fish quantity-microscopy of input)/throw in
1.4. natamycin is to the acute toxicity test of Ctenopharyngodon idellus
Test is carry out in the glass jar of 80L at volume, places healthy Ctenopharyngodon idellus 10 tail in each cylinder, and to add concentration be 50.0,60.0,70.0,80.0,90.0 and 100.0 mg L
-1control agent, adds up the dead quantity of Ctenopharyngodon idellus, and calculates the half lethal concentration of natamycin to Ctenopharyngodon idellus after 48h.Each concentration group does 3 repetitions, and test arranges a matched group.
2 results
Test data SPSS16.0 statistics software processes, and all parameters all represent by mean ± standard deviation (± SD).
2.1 external kill little melonworm test
Accompanying drawing 1 is shown in the killing in vitro effect of natamycin to little melonworm larva.From accompanying drawing 1: natamycin has stronger killing action to little melonworm, and strengthen gradually along with its killing action of increase of concentration, he is to little melonworm larva 10 min-, 1 h-, 2 h-, 3 h and 4 h-median lethal valid density (EC
50) concentration of (95% CI) is respectively 17.8 (17.5-18.1), 13.4 (13.2-13.6), 12.8 (12.5-13.1), 12.8 (12.4-13.2) and 10.9 (10.7-11.1) mg L
-1, its concentration is 25.0 mg L
-1time, be 100% to the killing rate of little melonworm.
Natamycin to the killing in vitro effect of little melonworm encapsulation in table 1.As shown in Table 1: natamycin is 25.0 mg L in concentration
-1encapsulation does not divide, and does not hatch larva yet, and it is 100% to the killing rate of encapsulation.Its concentration is 20.0,15.0 and 10.0 mg L
-1time 83.3%, 56.7% and 26.7% is respectively to the killing rate of encapsulation, and 5.0 mg L
-1concentration group does not act on substantially to encapsulation, and whole encapsulation all hatches out.Its 4 h-EC
50(95% CI) is 13.8 (13.5-14.1) mg L
-1.
Insecticidal test in the body of 2.2 natamycins
Natamycin to insecticidal test result in the body parasitized on Ctenopharyngodon idellus as table 2.As shown in Table 2: natamycin has stronger killing action to little melonworm, use after medicine, the little melonworm quantity on Ctenopharyngodon idellus body surface and the gill obviously reduces, and its drop after the incubation rate of the larvaes of encapsulation obviously reduce.When its concentration is 20.0 mg L
-1time, the mortality rate of Ctenopharyngodon idellus is 36.6%, on its gill and the little melonworm number of fin ray be 65.5 ± 14.0.And the Ctenopharyngodon idellus mortality rate of matched group is 86.7%, on the gill and fin ray, the quantity of little melonworm is 208.5 ± 35.0.Result shows that natamycin has the test fishing gear of little melonworm to have stronger protective effect to infection.
2.3 natamycins are to the acute toxicity test of Ctenopharyngodon idellus
Concentration is 50.0 mg L
-1after natamycin medication 48h, Ctenopharyngodon idellus mortality rate is 0, and Ctenopharyngodon idellus is active normal, also no abnormality seen reaction.Concentration is 100.0 mg L
-1namely after natamycin medication 4h there is rapid breathing in Ctenopharyngodon idellus, and the abnormal responses such as jump, after 48h, Ctenopharyngodon idellus is all dead.Natamycin is to the 48h LD of Ctenopharyngodon idellus
50be 72.8 mg L
-1(its 95% confidence interval is 70.1-75.5 mg L
-1).
Table 1 natamycin is to killing in vitro effect (the mg L of little melonworm encapsulation
-1)
Note: different letter representation significant difference (p < 0.05)
Table 2 natamycin is to insecticidal effect in the body of little melonworm
Note: different letter representation significant difference (p < 0.05)
two, natamycin kills the pharmacy in vitro test that parasitic Fish stimulate cryptonucleus insect
1 materials and methods
1.1 test fishes
Severe infections stimulates the Carnis Pseudosciaenae of cryptonucleus insect from Fuding, Fujian Province seed base.
Test medication is the control agent according to embodiment 1.
1.2 external insecticidal tests
Being stimulated by severe infections the Carnis Pseudosciaenae of cryptonucleus insect to put into volume is 1m
3in drum, discharge water with brush brush drum head gently in the next morning, collect the stimulation cryptonucleus insect of bottom.And with suction pipe, beaker is put in ripe polypide collection, a part of polypide is used for the cultivation of larva, and a part is used for encapsulation test.
1.2.1 natamycin is tested the killing in vitro of cryptocaryon irritans larva
Test is carried out on 24 porocyte culture plates, adds control agent (10,15,20, the 25 and 30 mg L of about 2ml variable concentrations in each cell hole
-1) and about 100 larvas, after medication, 15min, 1h, 2h, 3h, 4h microscopy is observed once, adds up the larval mortality in each hole after 4h.Test arranges dimethyl sulphoxide control group and fully sudden and violent gas Natural Water matched group, and often group test in triplicate.
Dead larvae criterion: polypide cilium does not move, and kytoplasm does not flow, and after birth breaks, nuclear fragmentation.
1.2.2 natamycin is tested the killing in vitro of encapsulation
Test is carried out on 24 porocyte culture plates, adds medicinal liquid (10,15,20, the 25 and 30 mg L of about 2ml variable concentrations in each cell hole
-1) and 30 encapsulations, after medication, 4h microscopy is observed once, adds up the stimulation cryptonucleus insect encapsulation mortality rate in each hole after 4h.Test arranges dimethyl sulphoxide control group and fully sudden and violent gas Natural Water matched group, and often group test in triplicate.
The dead criterion of encapsulation: encapsulation does not divide or encapsulation does not hatch larva.
2 results
Test data SPSS16.0 statistics software processes, and all parameters all represent by mean ± standard deviation (± SD).
The external insecticidal test of 2.1 natamycins
Accompanying drawing 2 is shown in the killing in vitro effect of natamycin to larva.From accompanying drawing 2: natamycin has stronger killing action to cryptocaryon irritans larva, along with its killing action of increase of concentration strengthens gradually, he is to larva 2 h-, 3 h-and 4h-median lethal valid density (EC
50) concentration of (95% CI) is respectively 13.2 (13.0-13.4), 12.5 (12.4-12.6) and 10.6 (10.3-10.9) mg L
-1.
Natamycin to the killing in vitro effect of little melonworm encapsulation in table 3.As shown in Table 3: natamycin is 30.0 mg L in concentration
-1encapsulation does not divide, and does not hatch larva yet, and it is 100% to the killing rate of encapsulation.Its concentration is 25.0,20.0,15.0 and 10.0 mg L
-1time 94.4%, 62.2%, 16.7% and 5.5% is respectively to the killing rate of encapsulation.Its 4 h-EC
50(95% CI) is 18.8 (18.4-19.2) mg L
-1.
Table 1 natamycin is to killing in vitro effect (the mg L stimulating cryptonucleus insect encapsulation
-1)
Note: different letter representation significant difference (p < 0.05)
Claims (9)
1. natamycin is being killed little melonworm and is being stimulated the application in cryptonucleus insect.
2. natamycin is being prepared the external little melonworm of killing aquatic animal and is being stimulated the application in cryptonucleus insect medicine.
3. a control agent for the external little melonworm of killing aquatic animal and stimulation cryptonucleus insect, it is characterized in that: the raw material comprising following weight percents: natamycin 5% ~ 25%, surfactant 5% ~ 10%, lytic agent 10 ~ 20%, all the other are solvent.
4. control agent according to claim 3, is characterized in that: described lytic agent is dimethyl sulfoxide or DMF or glacial acetic acid.
5. control agent according to claim 3, is characterized in that: described surfactant is tween.
6. control agent according to claim 5, is characterized in that: described surfactant is tween 20 or tween 80.
7. control agent according to claim 5, is characterized in that: described solvent is water.
8. the external little melonworm of killing aquatic animal and stimulate the preparation method of cryptonucleus insect control agent as claimed in claim 3, is characterized in that, carry out according to the following steps:
1) natamycin, surfactant, lytic agent and solvent is taken by described weight portion, for subsequent use;
2) lytic agent is added in natamycin, and be stirred to natamycin and dissolve completely;
3) surfactant and solvent are added step 2) stir in the mixed solution of gained, obtain control agent.
9. control agent as claimed in claim 3, its using method and consumption are: splash after dilute with water in fish pond, and using dosage is 15-25 mL/m
3.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108653261A (en) * | 2018-05-04 | 2018-10-16 | 浙江省淡水水产研究所 | Application of the 6- n-pentyloxies Rhein in prevention saprolegniasis and ich |
| CN110772510A (en) * | 2019-12-13 | 2020-02-11 | 浙江省淡水水产研究所 | Novel application of ligustilide |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101194614A (en) * | 2007-12-21 | 2008-06-11 | 华南农业大学 | Mixed pesticide preparation of tubatoxin and antibiotic fungicide |
| CN102293745A (en) * | 2011-08-25 | 2011-12-28 | 西北农林科技大学 | Natamycin nano-emulsion antifungal medicament and preparation method thereof |
-
2015
- 2015-02-16 CN CN201510082499.XA patent/CN104784194B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101194614A (en) * | 2007-12-21 | 2008-06-11 | 华南农业大学 | Mixed pesticide preparation of tubatoxin and antibiotic fungicide |
| CN102293745A (en) * | 2011-08-25 | 2011-12-28 | 西北农林科技大学 | Natamycin nano-emulsion antifungal medicament and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108653261A (en) * | 2018-05-04 | 2018-10-16 | 浙江省淡水水产研究所 | Application of the 6- n-pentyloxies Rhein in prevention saprolegniasis and ich |
| CN110772510A (en) * | 2019-12-13 | 2020-02-11 | 浙江省淡水水产研究所 | Novel application of ligustilide |
| CN110772510B (en) * | 2019-12-13 | 2022-07-19 | 浙江省淡水水产研究所 | Application of ligustilide |
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