CN103109810B - Rotifer killing medicine, and preparation method and application thereof - Google Patents
Rotifer killing medicine, and preparation method and application thereof Download PDFInfo
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- CN103109810B CN103109810B CN201110363769.6A CN201110363769A CN103109810B CN 103109810 B CN103109810 B CN 103109810B CN 201110363769 A CN201110363769 A CN 201110363769A CN 103109810 B CN103109810 B CN 103109810B
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to the biotechnical field, and concretely relates to a rotifer selective-killing medicine, and a preparation method and application thereof. The medicine is a polyol ester compound extracted from Celastraceae plants and having a structure represented by formula I shown in the specification. In the invention, the basic biological differences of animals and plants are compared, tissue organs and submicroscopic organelles which only exist in the animals and do not exist in microalgae are selected as main foci, the botanical medicine which can be degraded and has a small toxicity is screened, and the rotifer growth inhibition and spawning rate and hatching rate reduction are realized through selectively generating anorexia response, destroying the cell membranes and the organelle membranes of a digestion organ system and inhibiting the nerve-muscle excitatory junction potential. The rotifer killing medicine has the advantages of safe, rapidness and effective technology, and specificity of the rotifer killing and inhibition.
Description
Technical field
The invention belongs to biological technical field, one is killed wheel animalcule medicament and its preparation method and application specifically.
Background technology
Micro-algae (comprising the micro-algae of freshwater microalgae and seawater), as the important component part of primary productive force in biosphere, is unique bait of the whole process of growth of aquatic livestock (or specific developmental stage) in aquatic ecosystem or the bait of its food organisms.Meanwhile, it is floristic 50% that micro-algae approaches, and has the diversity of going out, complexity and particularity at aspects such as germplasm, ecological distribution, hereditary information, biochemical composition, metabolic pathways, and then determined the value such as its potential edible, nutrition, health care and pharmacy.Likely therefrom develop the special high added value bioactive ingredients of mankind's wilderness demand, structure (as carotenoid and derivative, AA, EPA and DHA equal altitudes unsaturated fatty acids such as beta carotene, astaxanthin, xanthophyll, and vitamin, polyol ester, amino-heterocycles, macrolide, polysaccharide and derivative thereof, nucleotide, differential protein and polypeptide etc.).In addition, the micro-algae of part can accumulate a large amount of greases under given conditions, and its oil content even exceedes oil crop seeds by using oil content.In addition micro-algae has the features such as high, the solid carbon ability of photosynthetic efficiency is strong, growth cycle is short, micro-algae production process and carbon dioxide discharge-reduction is coupled, its development of resources can be avoided striving and striving the advantages such as water with crops.Therefore, micro-algae is considered to the most desirable approach of mankind's regeneration biological diesel oil etc.
For these reasons, micro-algae has become the mankind continue the another important regeneration biological resources bank of successfully developing after microorganism, has been subject in recent years showing great attention to of domestic and international governments at all levels and scientific and technological circle.
But also should admit, in micro-algae industrialization production process, along with the progressively expansion of the scale of cultivation, often occur seriously restricting the particularly wheel animalcule pollution problem of harmful organisms of micro-algae living resources exploitation.Wheel animalcule has powerful fertility and ingestion ability, once there is wheel pest pollution in micro-algae cultivating system, several days time just, by micro-algae food light of cultivating, was caused heavy losses.
The harm of wheel animalcule in micro-algae incubation is mainly reflected in the following aspects: 1) breeding water body is endangered: wheel animalcule amount reproduction cause water body in dissolved oxygen sharply decline, the accumulation of its metabolite ammonia and toxin, causes water quality severe exacerbation; 2) to the harm of micro-algae: the wheel animalcule algae that eats in a large number, each wheel animalcule thousands of even up to ten thousand microalgae cells of ingesting every day.That is to say in micro-algae engineering culture, once pollute wheel animalcule in culture systems, will cause microalgae cell density to cause soon decline, and finally cause making whole cultivation unsuccessfully; 3) wheel animalcule breeding produces a large amount of ovum, remains in cultivating system, is difficult to thoroughly remove, and as pollution source to next time micro-algae incubation pollute, produce vicious circle, causing micro-algae to cultivate scale can not expand well, seriously limits micro-algae Biological Energy Industry development.
The technical method of in the past killing wheel animalcule is more, is mainly broad spectrum activity method, and selectivity is inadequate, mainly contains:
1) micro-algae is cultivated pretreatment technology method, such as ozone, ultraviolet ray, clorox, physical filtering etc., mainly micro-algae cultivated to material (water, fertilizer, gas) and cultivate facility and carry out disinfecting early stage, this is at present domestic and international current sterilization, purification method, mainly to reduce importing into and diffusion path of wheel animalcule and ovum thereof, this technology does not have selectivity, equally also micro-algae is produced to injury, be only applicable to cultivate pre-treatment, find that in incubation the micro-frond that infects wheel animalcule is and just can not be used for killing.
2) physical method in micro-algae incubation; as adopt the means such as micro porous filtration and nylon wire isolation to the control of algae kind purity; this technology can effectively be removed large-scale weed eater; but be difficult to eradicate for the very little food algae property wheel animalcule of individuality and ovum thereof; the rate of filtration is slower simultaneously, and the application in large-scale production is restricted.
3) chemical method in incubation, wheel animalcule and algae belong to eucaryote together, this characteristic makes still drug application (formaldehyde, ammoniacal liquor, hydrogen peroxide and clorox etc.) and reduce the technology such as pH simply of people, be that these methods all do not have selectivity, wheel animalcule and ovum thereof are not only controlled and removed to its processing procedure, can also cause serious harm to micro-algae simultaneously.
4) utilize heavy metal to control wheel animalcule, poisoning research to wheel animalcule mainly concentrates on Cu, Zn, Cd, Pb etc., find that this metalloid ion pair wheel animalcule has high toxicity, 24h half lethal concentration is in 1mg/L left and right, the method is just considered separately the impact on wheel animalcule, does not consider the impact on micro algae growth breeding and product quality.
5) organic phosphates, the agricultural chemicals such as carbamic acid class, the kill valid density that its effect depend on poisonous substance of these agricultural chemicals to wheel animalcule, does not consider the impact on micro algae growth breeding and products thereof quality equally.
In addition, the nineties in 20th century, single-minded optionally virus and fungi technical method are there is having, be Comps etc. is separated to virus and fungi (ocean double RNA virus and chain chytrid) in dead wheel polypide in plant, indoor infection wheel animalcule by experiment, proves that this viroids and fungi can cause wheel animalcule population quantity to reduce and bosom ovum rate reduces.Expect that following the method has selectivity, in microalgae mass is cultivated, may there is good application prospect, but at present still in laboratory development.
In sum; how to find that a kind of cost is strong compared with low, selectivity, easily degraded, little and be convenient to the medicine of large-scale promotion application to algae toxicity; kill wheel animalcule or suppress its growth; be the technology of controlling the required urgent solution of wheel animalcule harm in micro-algae incubation, also will determining the success or failure of micro-algae (comprising freshwater microalgae and seawater) industrialization.
Summary of the invention
The object of the invention is to provide one to kill wheel animalcule medicament and its preparation method and application.
For achieving the above object, the technical solution used in the present invention is: one is killed wheel animalcule medicament, and described medicament is polyol ester compound, shown in one,
Formula one.
Described medicament is the polyol ester compound of Celastraceae plant extract.
Kill the preparation method of wheel animalcule medicament: the polyol ester compound by Celastraceae plant extract is dissolved in completely in distilled water and is mixed with liquid mother liquor, then the mother liquor of configuration is added to and cultivate that in algae liquid, to make algae liquid herb liquid final dose be 0.1-0.32ppm.
Kill the application of wheel animalcule medicament: the polyol ester compound by Celastraceae plant extract is dissolved in distilled water, is mixed with liquid mother liquor completely, then the mother liquor of configuration is added to and cultivate in algae liquid, making algae liquid herb liquid final dose is 0.1-0.32ppm, after medication 4 hours start to play a role, 24 hour detection time drug effect maximum, wherein the medial lethal dose of 24 hours is 0.175ppm, and full lethal concentration is 0.32ppm.
Described liquid mother liquor is positioned over sealing in 4 DEG C of low temperature environments and preserves, and to prevent that drug volatilization from reducing concentration, and affects its drug effect, and liquid mother liquor was generally kept in 7 days.
The drug mechanism that kills wheel animalcule mainly comprises: 1) can reduce on mouthpart receptor the susceptibility of the neuroceptor to taking food generation effect, suppress the granting of the neuron signal that causes appetite, produce apocleisis reaction; 2) destroy digestive organs system cells film and organelle film, its structure is changed; 3) nerve-excitation of muscle junctional potential is produced to inhibitory action, and finally block current potential conduction, thereby make to be affected by the motion (breathing etc.) of Neuro mascular, axoneure sodium channel gate process is exerted an influence simultaneously; 4) suppress wheel animalcule and grow, reduce its spawning rate and incubation rate.
The present invention has advantages of:
Medicine of the present invention can directly use with aqua form in micro-algae incubation, optionally kills wheel animalcule.Through experiment repeatedly, the economy of this medicine is used less (concentration can drop to as 0.11ppm), and under this concentration, wheel animalcule individual reproduction power is significantly inhibited, and population proliferative speed significantly declines.Simultaneously, even according to using this medicine in the using dosages of 3 times (0.32ppm) situation, although can cause micro-algae chlorophyll content slightly to decline, the indexs such as cell proliferation speed, final micro algae biomass and photosynthetic performance are not exerted an influence substantially.Show the selectivity of wheel animalcule effect byer force, and do not affect micro algae growth.In actual production process, for the effect that ensures that wheel animalcule is killed in medication, can select relatively high dosage (0.32ppm).Adopt in addition medicament of the present invention not affect microalgae cell growth, only to wheel animalcule generation effect.As a kind of botanical components, to mammal low toxicity and the characteristic to non-target organism safety.Rat acute LD50 of passing through mouth is 0.68mg/Kg (being 0.68ppm), acute through skin LD50 > 2000mg/Kg.Above-mentioned data are according to the acute toxicity grading criteria of national agriculture chemical registration toxicological test method (G-B15670-1995), and its toxicity all belongs to low toxicity.
The present invention is directed to micro-algae autophyting growth, and the harmful organisms existence such as wheel animalcule and growth and breeding need to rely on the characteristic of micro-algae that constantly eats from the external world, the relatively strong medicine of screening selectivity, reaches selectivity and kills wheel animalcule or suppress its breeding.
Brief description of the drawings
Fig. 1 is the inhibitory action figure of medicament (ppm) to wheel animalcule population growth.
Embodiment
Embodiment 1
(1) according to the dosage of 100mg in every premium on currency body, medicine is dissolved in completely in distilled water and is mixed with liquid mother liquor (100ppm), liquid needs now with the current, to keep optimum medicine efficacy.If cannot realize with front interim preparation, liquid mother liquor should be positioned over to sealing in 4 DEG C of low temperature environments and preserve, to prevent that drug effect volatilization from reducing effect, the liquid mother liquor holding time is generally no more than 7 days.
(2) according to the volume of the cultivation algae liquid of infection wheel animalcule, get in proportion quantitative liquid medicine mother liquor and add in cultivation algae liquid, making liquid final dose is 0.32ppm.After adding liquid mother liquor, fully vibrate and mix algae liquid, realize drug effect maximization thereby liquid is evenly distributed in water body.Liquid joins in the cultivation algae liquid that infects wheel animalcule, and along with wheel animalcule constantly filters food activity, liquid will act on wheel animalcule mouthpart, digestive system and current potential conducting system etc. gradually, thereby it is killed.
(3) to the closed cultivating system of cultivating on a small scale, need not timing to shake up algae liquid; And the algae liquid that extensive ventilation is cultivated, because liquid has certain volatility, for ensureing that drug effect should suspend ventilation between liquid action period, adopts not timing all once stirring the increase of algae liquid method.
(3) after liquid effect 24h, get algae sample and observe, if individual without wheel animalcule survival, large-scale culture system can be recovered to continue ventilation, carries out normal frustule cultivation; If wheel animalcule contamination density is higher, after liquid effect 24h, in the micro-frond of cultivation system, may observe the survival wheel animalcule individuality that minority energy obviously weakens, this wheel animalcule speed of moving about is very slow, see under the microscope in its stomach and intestine almost without any food, now can select again proper extension to stop the supple of gas or steam the time, increase insecticidal effect.
The medicine that selectivity is killed wheel animalcule is the polyol ester compound from Celastraceae plant extract, and its chemical constitution is as follows.
Formula one.
Extracting method is edited referring to Wang Junru, and 2006, natural product extraction separates and authenticate technology.First section of the 2nd conventional extraction and separating and purifying technology brief introduction of chapter, the 2nd section of the 3rd chapter Comprehensive Experiment, by vegetable material drying and crushing, soaks with organic solvents such as ethanol, acetone, ether and chloroforms, and hot reflux is extracted.
Low concentration (< is less than 0.11ppm) medication effect See Figure, although the more not contrast decline of dispenser of wheel animalcule proliferative speed still has survival individual, and can continue breeding.Drug level is obvious dosage correlation with inhibition wheel animalcule reproductive effect, dosage is higher kills and to suppress wheel animalcule reproductive effect more obvious, lower than the processed group of 0.11ppm, in several days subsequently observe, still can breed, but the wheel animalcule activity in 0.11ppm processed group declines to a great extent, most appearance are dead, and observation in several days is subsequently almost all dead, and wheel animalcule proliferative speed is suppressed (referring to Fig. 1).
Embodiment 2
Difference from Example 1 is
According to the volume of the cultivation algae liquid of infection wheel animalcule, get in proportion quantitative liquid medicine mother liquor and add in cultivation algae liquid, making liquid final dose is 0.12ppm.After adding liquid mother liquor, fully vibrate and mix algae liquid, realize drug effect maximization thereby liquid is evenly distributed in water body.Liquid joins in the cultivation algae liquid that infects wheel animalcule, and along with wheel animalcule constantly filters food activity, liquid will act on wheel animalcule mouthpart, digestive system and current potential conducting system etc. gradually, thereby it is killed.After liquid effect 24h, get algae sample and observe, the wheel animalcule overwhelming majority is dead, and the swimming speed of indivedual active individuals is very slow, within several days subsequently, observes and does not find the wheel animalcule individuality of surviving, and wheel animalcule proliferative speed suppresses.
Embodiment 3
Difference from Example 1 is
According to the volume of the cultivation algae liquid of infection wheel animalcule, get in proportion quantitative liquid medicine mother liquor and add in cultivation algae liquid, making liquid final dose is 0.20ppm.After adding liquid mother liquor, fully vibrate and mix algae liquid, realize drug effect maximization thereby liquid is evenly distributed in water body.Liquid joins in the cultivation algae liquid that infects wheel animalcule, and along with wheel animalcule constantly filters food activity, liquid will act on wheel animalcule mouthpart, digestive system and current potential conducting system etc. gradually, thereby it is killed.After liquid effect 24h, get algae sample and observe, wheel animalcule is almost all dead, within several days subsequently, observes and does not also find that survival is individual.
Embodiment 4
Difference from Example 1 is
According to the volume of the cultivation algae liquid of infection wheel animalcule, get in proportion quantitative liquid medicine mother liquor and add in cultivation algae liquid, making liquid final dose is 0.30ppm.After adding liquid mother liquor, fully vibrate and mix algae liquid, realize drug effect maximization thereby liquid is evenly distributed in water body.Liquid joins in the cultivation algae liquid that infects wheel animalcule, and along with wheel animalcule constantly filters food activity, liquid will act on wheel animalcule mouthpart, digestive system and current potential conducting system etc. gradually, thereby it is killed.After liquid effect 24h, get algae sample and observe, do not find that survival is individual.In sum, 0.11ppm is economic dosage, but kills wheel animalcule effect for ensureing, can suitably improve Drug level to 0.32ppm.
Application examples
In July, 2011-August, it is stand-by that the liquid mother liquor 200ml that configures 100ppm taking distilled water is put in volume 4 DEG C of preservations in the glass reagent bottle of 250ml.
Take from the small ooecium algae of main collection separation and purification for cultivating algae strain, Initial seeding density is every milliliter of 20,000 frustules, (cultivates light intensity approximately 25 μ mol m through static cultivation in 3 days
-2s
-1, L: D=14: 10), algae liquid enters exponential phase of growth.According to 800ml algae liquid sub-bottle, totally 3 groups, 9 bottles.Wherein, 3 bottles is blank group; 3 bottles is wheel animalcule group, according to the density inoculating rotifer of every milliliter of 25 tails; Other 3 bottles is processed group, adds the medicinal mother liquor of 2.6ml (100ppm) in inoculating rotifer, and making the whole Drug level of liquid is 0.32ppm.
Carry out static cultivation by three groups respectively, not timing shaking flask during this time, after 24h, (Continuous Observation 7 days) observed in sampling, found that wheel animalcule group wheel animalcule well-grown, and the ability of moving about is strong, and bosom ovum rate is high, and processed group wheel animalcule is individual all dead.
In continuation incubation subsequently, wheel animalcule group is due to its effect of eating to small ooecium algae, and algae cell density, chlorophyll content, unit volume frustule dry weight biomass are all low compared with other group; And processed group algae cell density, net photosynthesis put oxygen speed, respiratory rate and chlorophyll integrated performance index etc. all with control group there was no significant difference, though its chlorophyll content slightly declines, final dry weight biomass is compared control group and be there is no significant change.
Above-mentioned example shows, this medicine is stronger to the selectivity of wheel animalcule effect, under drug concentration 0.32ppm effect, in 24h, can thoroughly kill the wheel animalcule polluting in micro-algae incubation, and frustule growth is not produced to obviously impact.
Claims (3)
1. one kind is killed the application of wheel animalcule medicament, it is characterized in that: the polyol ester compound by Celastraceae plant extract is dissolved in distilled water, is mixed with liquid mother liquor completely, then the mother liquor of configuration is added to and cultivate in algae liquid, making algae liquid herb liquid final dose is 0.1-0.32ppm, can effectively kill wheel animalcule after medication;
Described medicament is polyol ester compound, shown in one,
formula one.
2. application of killing wheel animalcule medicament claimed in claim 1, is characterized in that: described in to make algae liquid herb liquid final dose be 0.1ppm, after medication, can effectively kill wheel animalcule.
3. application of killing wheel animalcule medicament claimed in claim 1, is characterized in that: described liquid mother liquor is positioned over sealing in 4 DEG C of low temperature environments and preserves.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1109270A (en) * | 1994-03-30 | 1995-10-04 | 河南省科学院化学研究所 | Emulsion oil of Angulatin |
| KR100191901B1 (en) * | 1996-04-01 | 1999-06-15 | 박원훈 | Novel sesquiterpene ester compound |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1109270A (en) * | 1994-03-30 | 1995-10-04 | 河南省科学院化学研究所 | Emulsion oil of Angulatin |
| KR100191901B1 (en) * | 1996-04-01 | 1999-06-15 | 박원훈 | Novel sesquiterpene ester compound |
Non-Patent Citations (4)
| Title |
|---|
| A New Insecticidal Sesquiterpene Ester from Celastrus Angulatus;Shao-peng Wei等;《Molecules》;20090330;第14卷(第4期);第1396-1403页，第1400页Figure3 * |
| Shao-peng Wei等.A New Insecticidal Sesquiterpene Ester from Celastrus Angulatus.《Molecules》.2009,第14卷(第4期),第1396-1403页，第1400页Figure3. |
| 复方中草药渔用杀虫剂的研究;王先科;《湖南农业大学农业推广硕士学位论文》;20080615;第19-20页3.2.1 * |
| 王先科.复方中草药渔用杀虫剂的研究.《湖南农业大学农业推广硕士学位论文》.2008,第19-20页3.2.1. |
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