CN104285949B - The agent for killing of violent water flea - Google Patents
The agent for killing of violent water flea Download PDFInfo
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- CN104285949B CN104285949B CN201410450302.9A CN201410450302A CN104285949B CN 104285949 B CN104285949 B CN 104285949B CN 201410450302 A CN201410450302 A CN 201410450302A CN 104285949 B CN104285949 B CN 104285949B
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- presenegenin
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Abstract
The present invention discloses the agent for killing of a kind of violent water flea, it is characterized in that: described agent for killing is made up of the raw material of following weight percents: the presenegenin 30% of milkwort extract active constituent content >=90%, dispersant 16%, wetting agent 24%, carrier 30 as one kind %, does is described dispersant Morwet? D-400, does is described wetting agent Morwet? EFW, described carrier is diatomite.This is a kind of simple, convenient, low to cultivated animals toxicity, and can the disposable agent for killing thoroughly killing violent water flea.
Description
Technical field
The present invention relates to a kind of agent for killing of flea class, particularly the agent for killing of a kind of violent water flea.
Background technology
Violent water flea is the general designation of Crustachia Harpacticoida (Harpacticoida) some kinds, is the dominant groups in the mini-Frac test of ocean.In abalone, sea cucumber, the nursery of shrimp crab are produced, some kind amount reproduction, brings serious harmful animal to seed production in early stage.One is that violent water flea excessively multiplies in water body, not only consumes a large amount of oxygen, causes water hypoxia, and competes bait with cultivation Bao Miao, seedling, and simultaneously due to the shortage of Micro Algae, ammonia nitrogen cannot be utilized, and causes cultivation seed poisoning; Two is violent water flea amount reproductions, all stings food injury, cause the mortality of seed rearing process to Bao Miao, seedling, the shrimp juvenile crab that casts off a skin.Therefore, aborning, be one of key link in seed rearing to killing of violent water flea.At present, in production, one is that not only production cost is high by blocking the filtration of breeding water body, nor the extensive cultivation of adaptation seed is to the requirement of big yield; Two is that the domestic medicine that mainly adopts is killed.Many employings agricultural chemicals such as chlorophos, chrysanthemum ester class carry out killing of violent water flea in seawater offspring pond, and these agricultural chemicals not only cause murder by poisoning to cultivation Bao, sea cucumber, shrimp, crab etc., and key also causes severe contamination to water environment, also threatens human food's safety.In current fish production, the impact of drug abuse on healthy fish cultivation, ecotope and human health receives great concern.Therefore aquatic products disease control not only will focus on the hidden danger eliminating fish quality, but also medicine will be avoided the pollution factor of water environment, the research emphasis that the generation that application green non-pollution fishing medicine controls disease will be the great Disease management of fishery from now on.
Summary of the invention
The present invention is the above-mentioned deficiency in order to solve existing for prior art, proposes a kind of simple, convenient, low to cultivated animals toxicity, and can the disposable agent for killing thoroughly killing violent water flea.
Technical solution of the present invention is: the agent for killing of a kind of violent water flea, is characterized in that: described agent for killing is made up of the raw material of following weight percents:
The presenegenin 30% of milkwort extract active constituent content >=90%,
Dispersant 16%,
Wetting agent 24%,
Carrier 30 as one kind %,
Described dispersant is MorwetD-400,
Described wetting agent is MorwetEFW,
Described carrier is diatomite.
Compared with the existing technology, tool has the following advantages in the present invention:
Chinese herbal medicine is easily degraded at occurring in nature mostly as natural products, low to non-target organism toxicity, not easily produce drug resistance, therefore, use herbal control aquatic products disease, to protection water environment, produce green aquatic product and have great importance.Medicinal plant polygala root (PolygalatenuifoliaWilld.) is subordinate to Polygalaceae (Polygalaceae) rogation flower (Polygala).Presenegenin (tenuigenin) is the main active in polygala root, from payta skin by extraction and isolation and hydrolysis obtain.The present invention is then for presenegenin has excavated new purposes, open up new application, utilize the violent water flea agent for killing that presenegenin is prepared from for principle active component, for material culture and the equal nonhazardous effect of breeding water body, it is a kind of nuisanceless green fishing medicine, and it disposablely thoroughly can kill the violent water flea in breeding water body, and effect is very remarkable; Simultaneously its operation and preparation also all very easy, be particularly suitable for applying in the art, its market prospects are very wide.
Embodiment
The specific embodiment of the present invention will be described below.
Presenegenin is to sea cucumber acute toxicity experience
1 materials and methods
1.1 material
1.1.1 test sample
Test presenegenin purity used to provide for 90%(is prepared by Animal Science And Technology aquatic products system of Xibei Univ. of Agricultural & Forest Science & Technology laboratory).Accurately take 1.0g presenegenin monomer, fully dissolve by deionized water, constant volume is in 100mL volumetric flask, and being made into concentration is 10.0mg/mL, and 4 DEG C of Refrigerator stores are for subsequent use.
1.1.2 experimental animal
Testing sea cucumber used chooses in Dalian HaiBao Fishery Company Ltd. cultivation base, and physique is healthy and strong, and specification is even, average weight 1.47g.In water tank, support about 7d temporarily, throw something and feed every day 1 time.Toxicity test is carried out again after the condition of conforming.
1.1.3 test apparatus
One of ALC-1100.2 percentage electronic balance, Beijing Sai Duolisi instrument system Co., Ltd; KQ-500E type ultrasonic cleaner; 5 ~ 10 μ L, 20 ~ 200 μ L pipettor: VoluMate.MadeinGermany etc.
1.2 test method
1.2.1 experimental condition
Adopt hydrostatic formula test method(s), each plastic cistern (specification 60 × 60 × 80cm) adds filtered seawater 100L, pH7.9, and controlling water temperature is 16 ± 1 DEG C.First add after test liquid fully mixes and add test use sea cucumber again, every water tank puts 100 in a suitable place to breed.After putting sea cucumber in a suitable place to breed, require to keep dissolved oxygen in water at more than 5mg/L.In order to keep constant drug concentration, change water dosing again every 24h.
1.2.2 the setting of trial drug concentration
First, repeatedly carry out on trial test basis, determine the approximate range of formal test liquor strength, namely test the liquor strength (0.05mg/L) that death does not occur in the sea cucumber liquor strength (0.2mg/L) of being all poisoned to death in 24h and 96h.Within the scope of liquor strength 0.2 ~ 0.05mg/L, insert seven concentration tests groups by log concentration equal difference (tolerance d=0.02mg/L) and set up a blank group, carrying out formal test, under observing each concentration, test the situation of being poisoned to death of sea cucumber.Test repetition 3 times.
Duration of test statistics tests the death condition of sea cucumber in 12h, 24h, 48h and 72h under each drug concentration.Duration of test should be kept quite, and avoids any interference to test sea cucumber as far as possible.When the lethality of control group is less than 5%, in each time period, the lethality of test sea cucumber can correct; If the lethality of control group is greater than 5%, then should correct, updating formula adopts Abbott formula; If control group lethality reaches more than 20%, then, after searching reason, re-start toxicity test.
Abbott formula: p=p '-C/1-C
P: corrected mortality, i.e. sheerly drug-induced lethality
P ': experimental group lethality, i.e. natural cause and drug-induced lethality
C: control group lethality, the i.e. lethality that causes of natural cause
1.2.3 presenegenin minimum lethal concentration scope, median lethal concentration (LC
50)
The death condition of observed and recorded test sea cucumber in 12h, 24h, 48h and 72h, test sea cucumber each time period starts to occur that dead least concentration and next concentration are the minimum lethal concentration scope of this time period, and calculates lethality rate.Utilize return law of the straight line, use linear regression equation
(
the log concentration of-drug concentration;
the probability unit that-lethality converts to;
,
be respectively intercept and the slope of straight line.), obtain
with
,
with
computing formula be:
: the group number of drug concentration (lethality be 0 and 100% concentration do not count)
After obtaining a, b, just can determine regression equation, and then by the probability unit of 50% lethality
bring equation into, obtain
, get its antilogarithm, just draw the LC in this time
50, and calculate its fiducial limit of 95%, formula is:
LC
5095% fiducial limit
In formula:
=
N: for examination animal number (lethality be 0 and 100% concentration do not count)
X:LC
50the logarithm of value
1.2.4 the calculating of presenegenin safe concentration
According to Turbell formulae discovery safe concentration: safe concentration=
2 results and analysis
2.1 presenegenin median lethal concentration (LC
50)
2.1.1 each experimental concentration of presenegenin in different time to sea cucumber intoxicating situation
The sea cucumber death condition of each trial drug concentration in 12h, 24h, 48h, 72h is in table 1.In the high concentration experimental group that drug concentration is 0.18mg/L, clearly, after medication there is Novel presentation in about 3h in sea cucumber toxic reaction, sea cucumber climbs out of the water surface along tank wall, is shed in water body subsequently, and adhesive ability reduces even to be lost, part tells intestines, last dead, change skin.Be in the low concentration experimental group of 0.06mg/L in concentration, sea cucumber performance is quieter, activity without exception.
Table 1 presenegenin is to the result of being poisoned to death of sea cucumber
2.1.2 minimum lethal concentration scope, median lethal concentration (LC
50)
The death condition of statistics variable concentrations test liquid in 12h, 24h, 48h and 72h, the results are shown in Table 2.
Each experimental concentration is got its logarithm, obtain its log concentration, and its probable value that the percentage that each time period test sea cucumber is poisoned to death is tabled look-up to obtain, the probable value of the drug concentration logarithm of each time period and the lethality of correspondence thereof is substituted into formula, obtain regression equation, the median lethal concentration (LC of each time period can be calculated
50), and its LC
5095% fiducial limit.
Table 2 presenegenin is to the acute toxicity result of sea cucumber
The LC of table 3 each time period
50and the fiducial limit of 95%
As shown in Table 3, in 24h, the regression equation of gained is: y=-3.35+22.49x, LC
50=0.15mg/L, 95% be crediblely limited to 0.14 ~ 0.17mg/L; In 48h, the regression equation of gained is:
Y=-3.79+33.86x, LC
50=0.11mg/L, 95% be crediblely limited to 0.10 ~ 0.12mg/L; In 72h, the regression equation of gained is: y=-5.05+54.29x, LC
50=0.09mg/L, 95% be crediblely limited to 0.08 ~ 0.10mg/L.
2.1.3 safe concentration result of calculation
The safe concentration of presenegenin to sea cucumber is gone out according to Turbell formulae discovery.
Safe concentration=
=(0.11 × 0.3) ÷ (0.15 ÷ 0.11)
2=0.02mg/L
From result display, presenegenin is not very high to the safe concentration of sea cucumber, but in reality, concentration is when 0.06mg/L, and the sea cucumber phenomena of mortality do not occur 48h.Consider, presenegenin still has good promotional value as violent water flea agent for killing.
Presenegenin is to violent water flea killing effect
1 materials and methods
1.1 experimental animal
Test violent water flea used and be collected in waters, Dalian HaiBao Fishery Company Ltd. cultivation base.
1.2 trial drug
Testing presenegenin purity used is 90%.(presenegenin is that payta skin is separated obtains through extraction, multistage silica gel column chromatography.Prepared by Xibei Univ. of Agricultural & Forest Science & Technology's Animal Science And Technology aquatic products system fish disease laboratory and provide.）
1.3 test method
Use the conical flask that volume is 300mL, contain into abundant aeration seawater 100mL, trace inflation, keeps water temperature 16 ± 1 DEG C, pH7.8 ~ 7.9.During on-test, determine the approximate range of formal test liquor strength, namely test in liquor strength (0.032mg/L) and 48h that violent water flea is all poisoned to death in 12h and dead liquor strength (0.0005mg/L) does not occur.Within the scope of liquor strength 0.2 ~ 0.05mg/L, insert six concentration tests groups by log concentration geometric ratio and set up a blank group, carrying out formal test.Join respectively in water body according to the concentration that above-mentioned test is set up, stir.Often group puts into the violent water flea 300 collected subsequently.Then respectively 6,12,24,48,72h time, at the bottom of sucking-off conical flask, dead violent water flea, puts into culture dish and observes counting.Violent water flea sinks to bottom culture dish does not have movable death of thinking, sucking-off dead individuals, movable violent water flea is put back in former conical flask.Test repetition 3 times, and establish control group.
The violent water flea quantity of death at the bottom of setting-up time statistics bottle, calculating mean value, and contrast with control group, calculate killing rate, evaluate killing effect with this.
Result
Presenegenin kills violent water flea Be very effective, and as shown in Table 4, when concentration is from 0.001mg/L, through 72h, the killing rate of violent water flea reaches 100%; When the concentration of presenegenin is from 0.004mg/L, the killing rate of 48h to violent water flea reaches 100%, and in control group, the 48h lethality of violent water flea is lower than 5%.
Table 4 presenegenin 48h kills activity to violent water flea
The effect test of the violent water flea agent for killing of presenegenin
1 materials and methods
1.1 material
1.1.1 experimental animal
Test is chosen in Hypon cultivation base, Dalian with sea cucumber, and physique is healthy and strong, and specification is homogeneous, with above-mentioned test same size, and average weight 1. [a1] g.In water tank, support about 7d temporarily, throw something and feed every day 1 time.
Test violent water flea used and be collected in waters, Hypon cultivation base, Dalian.
1.1.2 for reagent product
Containing the violent water flea agent for killing of 30% presenegenin, prepared by Xibei Univ. of Agricultural & Forest Science & Technology's Animal Science And Technology aquatic products system fish disease laboratory and provide.
1.1.3 instrument and equipment
XH-B vortex mixer: healthy medical apparatus Co., Ltd of Jiangyan City; BX41 light microscope: OLYMPUS.MadeinJapan; Satorius ten thousand/electronic balance; Aquarium and temperature regulating device.
Test method
Adopt hydrostatic formula test method(s), each plastic cistern (specification 30 × 30 × 40cm) adds filtering sea 20L, pH7.9, and controlling water temperature is 16 ± 1 DEG C.During on-test, first according to test set up concentration (0,0.007,0.008,0.009,0.01,0.011,0.012,0.013g/m
3) join respectively in water body, stir.Often group adds test with sea cucumber 20 and violent water flea about 4000, and process of the test requires that maintenance dissolved oxygen in water is at more than 5mg/L.Add up violent water flea killing rate and sea cucumber lethality after 48h, sea cucumber statistics lethality method as previously mentioned.Violent water flea statistics lethality method is: during sampling, after stirring abundant mixed water body, get 1L water immediately at random in beaker, and the dead violent water flea quantity of statistics, approximately calculates the violent water flea lethality often organizing test.Test repetition 3 times, and establish control group.
2 results
The violent water flea agent for killing of 30% presenegenin, that kills violent water flea at 48h the results are shown in Table 5.As can be seen from Table 5, when concentration is 0.007g/m
3time, killing rate is 90.3%; When concentration is greater than 0.01g/m
3time, its killing rate can reach 100%; (0.007-0.013g/m within the scope of experimental concentration
3), all do not occur intoxicating phenomenon in test sea cucumber 48h, in control group there is death in violent water flea from 6h, and in 48h, lethality is less than 5%.Above result shows that said preparation is the good and biological fishing medicine with applications well prospect to sea cucumber safety of a kind of violent water flea killing effect.
Table 5 contains 30% presenegenin violent water flea agent for killing at 48h to violent water flea killing effect
Essence for a better understanding of the present invention.The specific embodiment provided below in conjunction with inventor is to describe the violent water flea agent for killing of presenegenin disclosed by the invention in detail, and these are only the good embodiments of the present invention, but are not limited to these embodiments.
Embodiment 1
Take presenegenin 28g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 14g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 20g, diatomite 38g, fully mix, obtain the violent water flea agent for killing of presenegenin.
Embodiment 2
The violent water flea agent for killing of presenegenin, mass percent is: presenegenin 26g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 12g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 24g, diatomite 38g.
Embodiment 3:
The violent water flea agent for killing of presenegenin, mass percent is: presenegenin 32g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 18g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 26g, diatomite 24g.
Embodiment 4:
The violent water flea agent for killing of presenegenin, mass percent is: presenegenin 34g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 20g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 25g, diatomite 21g.
Embodiment 5:
The violent water flea agent for killing of presenegenin, mass percent is: presenegenin 40g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 20g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 30g, diatomite 10g.
Embodiment 6:
The violent water flea agent for killing of presenegenin, mass percent is: presenegenin 42g, add dispersant (MorwetD-400, main component are alkylnaphthalene sulfonate formaldehyde condensation products) 25g, wetting agent (MorwetEFW, main component is the mixture of alkylnaphthalene sulfonate and anion wetting agent) 23g, diatomite 10g.
Claims (1)
1. an agent for killing for violent water flea, is characterized in that: described agent for killing is made up of the raw material of following weight percents:
The presenegenin 26-42% of milkwort extract active constituent content >=90%,
Dispersant 12-25%,
Wetting agent 20-30%,
Carrier 10-42%,
Described dispersant is MorwetD-400,
Described wetting agent is MorwetEFW,
Described carrier is diatomite.
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Citations (5)
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|---|---|---|---|---|
| CN1123176A (en) * | 1994-07-14 | 1996-05-29 | 埃米尔·弗拉克斯曼公开股份有限公司 | A process for the preparation of pesticide-poor concentrates of active components of plants |
| CN1623379A (en) * | 2004-12-14 | 2005-06-08 | 南京大学 | Wild bait with medicinal bait for eel and feeding method |
| CN1823916A (en) * | 2005-02-22 | 2006-08-30 | 徐东铭 | Extraction method of polygala refined total saponin |
| CN101828577A (en) * | 2010-03-16 | 2010-09-15 | 成都市翻鑫家科技有限公司 | Traditional Chinese medicine mosquito-repellent incense and preparation method thereof |
| CN102626114A (en) * | 2012-03-20 | 2012-08-08 | 杨俊奇 | Disease-curing insecticidal plant nutrient solution with pure traditional Chinese medicinal formula, and making method thereof |
-
2014
- 2014-09-05 CN CN201410450302.9A patent/CN104285949B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1123176A (en) * | 1994-07-14 | 1996-05-29 | 埃米尔·弗拉克斯曼公开股份有限公司 | A process for the preparation of pesticide-poor concentrates of active components of plants |
| CN1623379A (en) * | 2004-12-14 | 2005-06-08 | 南京大学 | Wild bait with medicinal bait for eel and feeding method |
| CN1823916A (en) * | 2005-02-22 | 2006-08-30 | 徐东铭 | Extraction method of polygala refined total saponin |
| CN101828577A (en) * | 2010-03-16 | 2010-09-15 | 成都市翻鑫家科技有限公司 | Traditional Chinese medicine mosquito-repellent incense and preparation method thereof |
| CN102626114A (en) * | 2012-03-20 | 2012-08-08 | 杨俊奇 | Disease-curing insecticidal plant nutrient solution with pure traditional Chinese medicinal formula, and making method thereof |
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