CN107027667B - Method and device for killing cryptocaryon irritans cysts by hydroxyl free radicals - Google Patents

Method and device for killing cryptocaryon irritans cysts by hydroxyl free radicals Download PDF

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CN107027667B
CN107027667B CN201710208886.2A CN201710208886A CN107027667B CN 107027667 B CN107027667 B CN 107027667B CN 201710208886 A CN201710208886 A CN 201710208886A CN 107027667 B CN107027667 B CN 107027667B
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active fragment
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culture pond
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CN107027667A (en
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白敏冬
王雅婧
余雯
张芝涛
林艳强
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Xiamen University
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01KANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K61/00Culture of aquatic animals
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish

Abstract

A method and a device for killing cryptocaryon irritans cysts by hydroxyl free radicals are provided with a water storage pool, a culture pool, a high-frequency high-voltage power supply, a modular oxygen active fragment generator, an online oxygen active fragment detector, a Venturi gas-liquid mixing and dissolving device, a liquid-liquid mixing and dissolving device, a TRO detector, a motor, an automatic rotating brush, a water pump, a valve, a gas pipeline and a liquid main pipeline; an air inlet of the modularized oxygen active fragment generator is externally connected with an oxygen source, and an oxygen active fragment air outlet of the modularized oxygen active fragment generator is sequentially connected with an online oxygen active fragment detector and an air suction port of the venturi air-liquid mixing and dissolving device; the water outlet of the water storage tank is connected with the water inlet of a water pump, the water outlet of the water pump is connected with the liquid inlet of a Venturi gas-liquid mixing and dissolving device, the liquid outlet of the Venturi gas-liquid mixing and dissolving device is connected with the liquid inlet of a liquid-liquid mixing and dissolving device, and the liquid outlet of the liquid-liquid mixing and dissolving device is sequentially connected with a TRO detector and the water inlet of a culture pond; the middle of the culture pond is provided with an automatic rotating brush controlled by a motor; the water outlet at the bottom of the culture pond is externally connected with seawater.

Description

Method and device for killing cryptocaryon irritans cysts by hydroxyl free radicals
Technical Field
The invention relates to the fields of plasma chemistry, seawater aquaculture technology, marine environmental engineering and the like, in particular to a method and a device for killing cryptocaryon irritans cysts by hydroxyl radicals.
Background
Cryptocaryon irritans are ciliates, cause white spot disease of marine fishes, easily explode in water with high culture density, poor fluidity and rich organic matter content, usually parasitize skin, fins, gills and other parts of tropical and subtropical marine fishes to cause organ damage and dysfunction of the fishes, cause abnormal activity, epithelial hyperplasia, dyspnea and mechanical damage of the diseased fishes, cause massive death of the fishes and cause great harm to the marine culture industry.
The life history of cryptocaryon irritans includes 4 stages of "trophozoite", "cystogenic" and "larval". The trophozoite is formed by growth and development of larvae parasitizing in epithelial tissues of diseased fish after the diseased fish is infected by the larvae, and the growth period is 3-7 d; after maturation, the diseased fish in the morning escape from the host and develop into an encapsulating precursor; after 8-12 h, the cilia of the cyst precursor are removed, and the cyst precursor is adhered to the underwater support to form a hard shell cyst; the cysts are subjected to a series of asymmetric binary divisions for 3-20D to form offspring larvae, each cyst can release 200-400 larvae, the larvae swim in water and infect host fishes to form trophozoites to carry out the next life cycle (but the study on generation, preservation and control of Cryptocaryon irritans (Cryptocaryon irritans) is proved and stimulated [ D ] river south university, 2006).
With the development of economy in China, the demand of people on marine products is gradually increased, the market demand can not be met by only depending on the traditional marine fishing, and the marine fish culture industry is rapidly developed. The cryptocaryon irritans cause has the characteristics of high propagation speed, high death rate, difficult control and the like, thereby causing great harm to the mariculture industry. The cryptocaryon irritans in the life history stage of the cysts have a plurality of layers of cyst walls, compact substances are arranged between the layers, the outer layer of the cysts is difficult to break, common medicines are difficult to enter the inside of the cysts, and the cysts cannot be effectively killed by the traditional method. The key to preventing cryptocaryon irritans is to kill cryptocaryon irritans cysts and prevent them from developing and releasing more larvae to repeatedly infect the cultured fish. How to kill the cryptocaryon irritans cysts with high efficiency, energy conservation and environmental protection receives more and more attention. At present, some physical and chemical methods are commonly adopted in mariculture production to kill cryptocaryon irritans cysts. The physical methods mainly comprise low-pressure ultraviolet irradiation, fresh water soaking of cysts, alternate culture and the like, but the effect is not obvious and the cysts cannot be effectively killed. The chemical method comprises the steps of splashing copper sulfate, formalin, potassium permanganate solution and the like into the sick culture pond, the methods have certain killing effect on cysts but cannot achieve effective and complete killing, most cysts can still continue to develop and release larvae to infect host fishes, and the problems of drug residues, toxicity to fishes, enhanced drug resistance of cysts, environmental pollution and the like can also exist after long-term overdose. Lianxing et al used a method of laying cloth mats at the bottom of a culture pond to remove cysts to control cryptocaryon irritans disease (see Chinese patent CN 105532534A). How to kill the cryptocaryon irritans cysts with high efficiency, energy conservation and environmental protection receives more and more attention.
Disclosure of Invention
The invention aims to solve the problems of poor cyst killing effect, serious environmental pollution and the like of cryptocaryon irritans during outbreak period in the existing seawater aquaculture, and provides a method and a device for killing cryptocaryon irritans cysts by hydroxyl radicals, which can kill the cryptocaryon irritans and prevent the cultured fishes from being repeatedly infected by more larvae released by the development of the cysts.
A device for killing cryptocaryon irritans cyst by hydroxyl free radical is provided with a water storage tank, a culture tank, a high-frequency high-voltage power supply, a modular oxygen active fragment generator, an online oxygen active fragment detector, a Venturi gas-liquid mixing device, a liquid-liquid mixing device, a TRO detector, a motor, an automatic rotating brush, a water pump, a valve, a gas pipeline and a liquid main pipeline; the air inlet of the modularized oxygen active fragment generator is externally connected with an oxygen source, and the oxygen active fragment air outlet of the modularized oxygen active fragment generator is sequentially connected with the online oxygen active fragment detector and the air suction port of the Venturi air-liquid mixing and dissolving device; the water outlet of the water storage tank is connected with the water inlet of a water pump, the water outlet of the water pump is connected with the liquid inlet of a Venturi gas-liquid mixing and dissolving device, the liquid outlet of the Venturi gas-liquid mixing and dissolving device is connected with the liquid inlet of a liquid-liquid mixing and dissolving device, and the liquid outlet of the liquid-liquid mixing and dissolving device is sequentially connected with a TRO detector and the water inlet of a culture pond; an automatic rotating brush controlled by a motor is arranged in the middle of the culture pond; the water outlet at the bottom of the culture pond is externally connected with seawater.
An oxygen valve and a gas flowmeter can be sequentially arranged between the oxygen source and the modular oxygen active fragment generator; an oxygen-containing active fragment gas valve and an oxygen-containing active fragment gas flowmeter can be sequentially arranged between the online oxygen active fragment detector and the Venturi gas-liquid mixing and dissolving device; a main pipeline liquid valve can be arranged between the water storage tank and the water pump, and a liquid flow meter can be arranged between the water pump and the Venturi gas-liquid mixing and dissolving device; an outward liquid discharge valve can be arranged between the culture pond and the external seawater; a water inlet main pipeline sampling valve can be arranged between the TRO detector and the culture pond, and an external drainage sampling valve can be arranged between the culture pond and external seawater.
A method of killing cryptocaryon irritans cysts by hydroxyl radicals comprising the steps of:
1) 7-11 months in summer every year, when the water temperature is 25-30 ℃ and cryptocaryon irritans is easy to outbreak, if the feeding of the cultured fishes is reduced, the bodies rub the wall or the bottom of the pond, the mucus on the body surface is increased, obvious white spots are formed, and part of the fries die; observing white spots on the surface of the dead fish body by using a microscope, and determining that the fish body is black round or pear-shaped, namely the cryptocaryon irritans disease can be confirmed; in the morning of 10: 00 after a large number of cysts fall off from the diseased fish, fishing out all the diseased fish in the diseased culture pond to other disease-free clean culture ponds, reducing the water level in the diseased culture pond, and attaching a large number of cryptocaryon irritans cysts to the bottom of the diseased culture pond;
2) opening a switch of the modular oxygen active fragment generator, opening an oxygen valve to introduce oxygen, and starting a high-frequency high-voltage power supply to ensure that the oxygen passing through the modular oxygen active fragment generator is quickly ionized and dissociated into gaseous oxygen active fragments; opening an oxygen-containing active fragment valve, detecting the concentration of the gaseous oxygen active fragment by an online oxygen active fragment concentration detector, and then entering a Venturi gas-liquid mixing and dissolving device;
3) opening a water pump and a main pipeline liquid valve, and injecting the clean seawater stored in the water storage tank into the Venturi gas-liquid mixing and dissolving device through the water pump; the gaseous oxygen active fragments entering the Venturi gas-liquid mixing and dissolving device in the step 2) and seawater are fully mixed and dissolved in the Venturi gas-liquid mixing and dissolving device and the liquid-liquid mixing and dissolving device to generate high-concentration hydroxyl radical solution, and the high-concentration hydroxyl radical solution enters the culture pond after passing through a TRO detector through a pipeline;
4) after the hydroxyl radical solution is injected into the culture pond, the bottom of the culture pond is soaked to stimulate cryptocaryon cyst for 0.5-2.0 h, the hydroxyl radical solution oxidizes a weak area of the cyst wall to cause the cyst wall to generate holes so as to enter the cyst, substances forming a basic cell structure and functional substances generate irreversible reactions, the irreversible reactions comprise amino acid oxidative decomposition, DNA chain breakage and the like, the cyst is dead, further development and larva release cannot be realized, and the residual hydroxyl radical is decomposed into O2And H2O; the middle of the culture pond is provided with an automatic rotating brush controlled by a motor, and the automatic rotating brush automatically rotates to clean the pond capsule after being started; and after rotating for 10min, opening an outward liquid discharge valve to discharge dead cysts in the culture pond along with the seawater.
In the step 1), the culture pond is a cylinder-like open type cement pond with the radius of 2m, the depth of 1.5m and the total volume of about 20m3. A water inlet is arranged above the side surface of the culture pond, the bottom of the pond is a horizontal smooth bottom of the pond, an automatic rotating brush controlled by a motor is arranged in the center of the pond, a rotating center shaft is a stainless steel pipe, a brush plate is arranged at a position 0.13m away from the bottom of the stainless steel pipe, the brush plate is a polypropylene smooth brush plate with the length of 3.8m and the width of 0.3m, and nylon wire bristles with the length of 0.15m are distributed on the brush plate; after the motor is started, the brush can rotate along the stainless steel pipe of the center shaft at the rotating speed of 10r/min, and the bottom of the side surface of the pool wall is provided with a water outlet.
In the step 2), the power of the high-frequency high-voltage power supply is 150-300W, the oxygen gas inflow is 1.0-5.0L/min, oxygen is ionized and dissociated after being bombarded by high-energy electrons to form gaseous oxygen active fragments, and the gaseous oxygen active fragments mainly comprise O2 +、O(1D)、O2(a1g)、O2 -、O(3P) and O3The concentration of the particles can reach 50-300 mg/L; the pipeline is a polytetrafluoroethylene pipe.
In the step 3), the flow rate of the seawater is 10-50L/min, the concentration of the hydroxyl radical solution is regulated and controlled by the water temperature, the concentration of the oxygen active fragments, the equilibrium partial pressure of the oxygen active fragment-containing gas and the gas-liquid volume ratio, and the concentration of a total oxidant TRO is 2-30 mg/L; the water storage tank is a square closed cement tank with the total volume of about 20m3The stored seawater is clean seawater pretreated by sand filtration and the like.
In the step 4), the cryptocaryon irritans cysts are soaked in the hydroxyl radical solution for 0.5-2.0 h, after death of the cysts is determined by means of microscopic examination and the like, an automatic rotating brush switch is started, after rotation for 10min, the dead cysts in the culture pond are discharged along with seawater, the used liquid pipeline can be made of stainless steel pipes subjected to corrosion prevention treatment, and the treated culture pond can be used for continuously culturing fishes.
The invention ionizes and dissociates oxygen into high-concentration oxygen active fragments in a very narrow discharge gap by utilizing atmospheric pressure ionization discharge, and then generates hydroxyl radical solution efficiently and rapidly within 1s through gas-liquid mixing and solution-liquid mixing. The hydroxyl free radical solution acts on the weak area of the capsule wall to cause the weak area to generate holes so as to enter the capsule, and substances and functional substances forming the basic structure of the cell generate irreversible reactions such as amino acid oxidative decomposition, DNA chain breakage and the like, so that the capsule is dead, more larvae are released during development and the cultured fish is repeatedly infected, and the effect of treating the cryptocaryon irritans is achieved. The hydroxyl free solution can quickly kill microorganisms such as residual harmful bacteria and the like in the culture pond while killing cryptocaryon irritans cysts, thereby preventing and treating plant diseases and insect pests.
In the device for killing and stimulating the cryptocaryon irritans cyst by the hydroxyl free radicals, the concentration of the oxidant is regulated and controlled by water temperature, the concentration of oxygen active fragments, the equilibrium partial pressure of oxygen active fragment-containing gas and the gas-liquid volume ratio; the concentration and the soaking time of the hydroxyl radical solution are determined according to the disease condition and the number of the cysts attached to the bottom of the pool, and the hydroxyl radical solution with the concentration of 5.0-10.0 mg/L can efficiently kill the cryptocaryon irritans cysts in 0.5-2.0 h, so that the cryptocaryon irritans cannot continue to develop to form larvae.
The invention relates to a method for killing cryptocaryon irritans cysts by using hydroxyl free radicals (OH) generated by atmospheric pressure ionization discharge, which is mainly used for killing cryptocaryon irritans cysts in seawater aquaculture2O and O2No residual medicine, etc. The method and the device for killing cryptocaryon irritans cysts based on the hydroxyl radicals provide a new solution for preventing and treating cryptocaryon irritans in seawater aquaculture.
The invention has the technical effects and advantages that ① leads oxygen into the submicron Al with extremely narrow discharge gap of 0.1mm2O3In a miniaturized non-equilibrium plasma source which is smelted into a thin dielectric layer, the generation of high-concentration oxygen active particles by atmospheric pressure ionization discharge is realized; adopting modularized array plasma integrated source and random combination to obtain controllable and large-output oxygen active particles, and cooperating with water jet cavitation gas-liquid mixing and dissolving technology to obtain high-concentration hydroxylThe method comprises the steps of preparing a free radical solution, soaking an Cryptocaryon irritans capsule adhered to the bottom of a culture pond in a hydroxyl free radical solution with the concentration of 5.0-10.0 mg/L for 0.5-2.0 h, wherein the TRO concentration is 2.0-30.0 mg/L, ② can completely kill the Cryptocaryon irritans capsule adhered to the bottom of the culture pond, and prevent the Cryptocaryon irritans capsule from developing and releasing more larvae to repeatedly infect cultured fishes, ③ can sterilize and disinfect the culture pond while killing the Cryptocaryon irritans capsule to prevent diseases and insect pests, ④ can thoroughly remove the Cryptocaryon irritans capsule adhered to the bottom of the culture pond by using an automatic rotating hairbrush, and the dead capsule is directly discharged along with residual seawater in the culture pond, and ⑤ is simple to implement, low in operation cost, free of drug residue while efficiently killing the Cryptocaryon irritans capsule, free of toxicity to fishes, free of pollution to the environment, and provides a healthy and environment-friendly device for killing the Cryptocaryon irrita.
Drawings
FIG. 1 is a schematic structural diagram of an embodiment of the device for killing and stimulating Cryptocaryon irritans by hydroxyl radicals according to the present invention.
FIG. 2 shows the change in the morphology of the capsules before and after killing of hydroxyl radicals in an example of the present invention. In fig. 2, A, C is a control group capsule morphology photograph; B. d is a photograph of the encapsulation form when TRO is 5.27 mg/L.
Detailed Description
The invention is further explained with reference to the drawings. The scope of protection of the invention is not limited to the embodiments described.
Referring to fig. 1, the device for rapidly killing cryptocaryon irritans by hydroxyl radicals is provided with a water storage tank 1, a venturi gas-liquid mixing and dissolving device 5, a high-frequency high-voltage power supply 6, a modular oxygen active fragment generator 7, an online oxygen active fragment detector 8, a liquid-liquid mixing and dissolving device 9, a TRO detector 10, a motor 11, an automatic rotating brush 12 and a culture pond 13.
The water outlet of the water storage tank 1 is connected with the water inlet of a water pump 31, and a 1 st valve 21 is arranged between the water storage tank 1 and the water pump 31; the water outlet of the water pump 31 is connected with the liquid inlet of the Venturi gas-liquid mixing and dissolving device 5, and a 1 st flow meter 41 is arranged between the water pump 31 and the Venturi gas-liquid mixing and dissolving device 5; an air inlet of the modularized oxygen active fragment generator 7 is connected with an external oxygen source, and a 2 nd valve 22 and a 2 nd flow meter 42 are sequentially arranged between the modularized oxygen active fragment generator 7 and the oxygen source; an oxygen active fragment gas outlet of the modularized oxygen active fragment generator 7 is sequentially connected with an online oxygen active fragment detector 8 and a gas suction port of the Venturi gas-liquid mixing device 5, and a 3 rd valve 23 and a 3 rd flow meter 43 are sequentially arranged between the online oxygen active fragment detector 8 and the Venturi gas-liquid mixing device 5; the liquid outlet of the Venturi gas-liquid mixing and dissolving device 5 is connected with the liquid inlet of the liquid-liquid mixing and dissolving device 9; a liquid outlet of the liquid-liquid mixing and dissolving device 9 is sequentially connected with a TRO detector 10 and a water inlet of the culture pond 13, and a water inlet main pipeline sampling valve 24 is arranged between the TRO detector 10 and the culture pond 13; the middle of the culture pond 13 is provided with an automatic rotating brush 12 controlled by a motor 11; the bottom water outlet of the culture pond 13 is externally connected with seawater, and an external discharge 6 th valve 26 and an external discharge sampling valve 25 are arranged between the bottom water outlet of the culture pond and the seawater.
The method of hydroxyl radical killing of cryptocaryon irritans cysts is given below:
1) when cryptocaryon irritans are easy to outbreak when the water temperature is 25-30 ℃ in 7-11 months in summer every year, the cultured fishes have the conditions of reduced ingestion, friction of the pond wall or the pond bottom by the body, increase of mucus on the body surface, obvious white spots and death of part of fries; and observing white spots on the surface of the dead fish body by using a microscope, and determining that the fish body is in a black round shape or a pear-shaped solid shape, so that the cryptocaryon irritans disease can be confirmed. In the morning of 10: 00 after a large number of cysts fall off from the diseased fish, the diseased fish in the diseased fish culture pond 13 is all fished out to other disease-free clean culture ponds, the water level of the diseased fish culture pond 13 is reduced, and a large number of cryptocaryon irritans cysts are attached to the bottom of the diseased fish culture pond 13. And (3) opening the high-frequency high-voltage power supply 6 to enable excitation voltage to be generated outside the modular oxygen active fragment generator 7, simultaneously opening the valve 22, enabling high-purity oxygen to enter the modular oxygen active fragment generator 7 through the flow meter 42 to be ionized and dissociated to generate gas containing oxygen active fragments, enabling 1/15-1/10 gas containing the oxygen active fragments to enter the online oxygen active fragment detector 8 to detect the concentration of the oxygen active fragments, and enabling the rest 9/10-14/15 gas to enter the Venturi gas-liquid mixing and dissolving device 5. The power of the high-frequency high-voltage power supply is 150-300W, and the air inflow of oxygen is 1.0-5.0L/min; the gas flow of the oxygen active fragment concentration detector is 0.1-0.5 mL/min, and the oxygen active fragment concentration is 50-300 mg/L.
2) Opening the valve 21 and the water pump 31 to enable the seawater stored in the water storage tank 1 to enter the Venturi gas-liquid mixing and dissolving device 5 after passing through the flow meter 41; the water flow is determined according to the number of cryptocaryon irritans cysts to be treated and can be 10-50L/min; fully mixing seawater and the gas containing the oxygen active fragments prepared in the step 1) in a Venturi gas-liquid mixing and dissolving device 5 and a liquid-liquid mixing and dissolving device 9 to generate a high-concentration hydroxyl radical solution, wherein the concentration of the hydroxyl radical solution is regulated and controlled by water temperature, the concentration of the oxygen active fragments, the equilibrium partial pressure of the gas containing the oxygen active fragments and the gas-liquid volume ratio, and the total active oxidant concentration is detected by an online TRO detector 10 and then enters a culture pond 13, and is 2-30 mg/L.
3) Injecting the high-concentration hydroxyl radical solution generated in the step 2) into a culture pond 13 for 1h, wherein the water flow is 10-50L/min, and the water surface height is 0.1-0.5 m; soaking the cryptocaryon irritans cysts adhered to the bottom of the culture pond 13 in the hydroxyl radical solution for 0.5-2 h, inspecting the death condition of the cryptocaryon irritans cysts in a microscope microscopic examination mode, and judging the cryptocaryon irritans to be dead if the cysts are light in color and have no obvious split phase.
4) After the cryptocaryon irritans are soaked in the hydroxyl radical solution in the step 3) to be killed, the motor 11 is turned on, the automatic rotating brush 12 is rotated to work, and the brush rotates along the central axis at the rotating speed of 10 r/min; after 10min of rotation the motor 11 is switched off and the valve 26 is opened to allow the dead cysts to be discharged with the seawater directly into the offshore area.
Specific examples of killing cryptocaryon irritans cysts are given below:
experiment of killing Cryptocaryon irritans cysts by hydroxyl radical solution.
In 8 months, in some cultivation factories of Pagrus major on small hill path island in Xiamen city, the ingestion of fries is reduced, the body rubs the pool wall or pool bottom, mucus on the surface gill part is increased, obvious white spots are formed, and part of fries are dead. The water temperature of the culture pond is 27-28 ℃, the salinity is 25.6 per mill, the pH is 7.75, and the dissolved oxygen is 6.78 mg/L. Microscopic examination finds that a large number of black round or pear-shaped worm bodies are wrapped in mucus on the surfaces of diseased fishes; and the bottom of the culture pond has granular hard matters, and the cryptocaryon irritans disease is judged to appear. Collecting the cryptocaryon irritans cysts at the bottom of the pool for killing experiment, and representing the killing conditions of the cryptocaryon irritans cysts under different OH dosages. The number of cryptocaryon irritans cysts used in the experiment is 100, the discharge power is 200W, and the concentration of the needed hydroxyl radical solution is adjusted by adjusting the gas-liquid mixing ratio. Four experimental groups, namely a control group and three hydroxyl radical treatment groups with different concentrations are arranged, and the concentrations of the treatment groups are 1.32mg/L, 2.63mg/L and 5.27 mg/L. And (3) directly introducing the hydroxyl radical solutions with different concentrations into a beaker filled with the cryptocaryon irritans cysts, soaking for 2 hours, and observing the forms of the cryptocaryon irritans cysts before and after killing the hydroxyl radicals by using a microscope. As shown in FIG. 2, the capsules of the control group had complete shells and appeared as black pears or circles, and distinct cleavage phases were observed in the capsules; the capsules were placed in clean sea water at 25.2 ℃ for a period of time and found to hatch and release larvae. The cryptocaryon irritans cysts treated by the hydroxyl radical solution with the concentration of 5.27mg/L have complete cyst walls, but obviously lightened cyst color and no split phase; the treated cysts were placed in clean seawater at 25.2 ℃ and no further development of the cysts was observed to release larvae. The killing effect of the hydroxyl radical solution with different concentrations on the cryptocaryon irritans cyst is shown in table 1.
TABLE 1 Effect of hydroxyl radical solutions of different concentrations on cryptocaryon irritans cyst stimulation
Figure BDA0001260534590000061
The hydroxyl radical solution can oxidize the weak area of the capsule wall to generate holes and enter the capsule, the substances and functional substances forming the basic cell structure undergo irreversible reactions such as amino acid oxidative decomposition, DNA chain breakage and the like, so that the capsule is dead and cannot be further developed to release larvae, and the residual hydroxyl radicals are decomposed into O2And H2O, hydroxyl radical solution with concentration of 5.27mg/L can completely kill and stimulate the cryptocaryon irritans to encapsulate.

Claims (5)

1. A method for killing cryptocaryon irritans cysts by hydroxyl radicals is characterized in that a device for killing the cryptocaryon irritans cysts by the hydroxyl radicals is provided with a water storage tank, a culture tank, a high-frequency high-voltage power supply, a modular oxygen active fragment generator, an online oxygen active fragment detector, a Venturi gas-liquid mixing and dissolving device, a liquid-liquid mixing and dissolving device, a TRO detector, a motor, an automatic rotating brush, a water pump, a valve, a gas pipeline and a main liquid pipeline; the air inlet of the modularized oxygen active fragment generator is externally connected with an oxygen source, and the oxygen active fragment air outlet of the modularized oxygen active fragment generator is sequentially connected with the online oxygen active fragment detector and the air suction port of the Venturi air-liquid mixing and dissolving device; the water outlet of the water storage tank is connected with the water inlet of a water pump, the water outlet of the water pump is connected with the liquid inlet of a Venturi gas-liquid mixing and dissolving device, the liquid outlet of the Venturi gas-liquid mixing and dissolving device is connected with the liquid inlet of a liquid-liquid mixing and dissolving device, and the liquid outlet of the liquid-liquid mixing and dissolving device is sequentially connected with a TRO detector and the water inlet of a culture pond; an automatic rotating brush controlled by a motor is arranged in the middle of the culture pond; the water outlet at the bottom of the culture pond is externally connected with seawater; an oxygen valve and a gas flowmeter are sequentially arranged between the oxygen source and the modular oxygen active fragment generator; an oxygen-containing active fragment gas valve and an oxygen-containing active fragment gas flowmeter are sequentially arranged between the online oxygen active fragment detector and the Venturi gas-liquid mixing and dissolving device; a main pipeline liquid valve is arranged between the water storage tank and the water pump, and a liquid flow meter is arranged between the water pump and the Venturi gas-liquid mixing and dissolving device; an outward liquid discharge valve is arranged between the culture pond and the external seawater; a water inlet main pipeline sampling valve is arranged between the TRO detector and the culture pond, and an external drainage sampling valve is arranged between the culture pond and external seawater;
the method comprises the following steps:
1) in 7-11 months every year, when the water temperature is 25-30 ℃ and cryptocaryon irritans easily outbreaks, if the feeding of the cultured fishes is reduced, the bodies rub the wall or the bottom of the pond, the mucus on the body surface is increased, obvious white spots are formed, and part of the fry is dead; observing white spots on the surface of the dead fish body by using a microscope, and determining that the fish body is black round or pear-shaped, namely, determining that the cryptocaryon irritans disease appears; in the morning of 10: 00 after a large number of cysts fall off from the diseased fish, fishing out all the diseased fish in the diseased culture pond to other disease-free clean culture ponds, reducing the water level in the diseased culture pond, and attaching a large number of cryptocaryon irritans cysts to the bottom of the diseased culture pond;
a water inlet is arranged above the side surface of the disease breeding pond, the pond bottom is a horizontal smooth pond bottom, an automatic rotating brush controlled by a motor is arranged in the center of the pond, a rotating center shaft is a stainless steel pipe, a brush plate is arranged at a position 0.13m away from the bottom of the stainless steel pipe, the length of the brush plate is 3.8m, and the width of the brush plate is 0.3 m; the brush plate is a smooth brush plate made of polypropylene material, and nylon wire bristles with the length of 0.15m are distributed on the brush plate; after the motor is started, the automatic rotating brush rotates along the rotating central shaft stainless steel pipe at the rotating speed of 10r/min, and the bottom of the side surface of the pool wall is provided with a water outlet;
2) opening a switch of the modular oxygen active fragment generator, opening an oxygen valve to introduce oxygen, and starting a high-frequency high-voltage power supply to ensure that the oxygen passing through the modular oxygen active fragment generator is quickly ionized and dissociated into gaseous oxygen active fragments; opening an oxygen-containing active fragment valve, detecting the concentration of the gaseous oxygen active fragment by an online oxygen active fragment concentration detector, and then entering a Venturi gas-liquid mixing and dissolving device;
3) opening a water pump and a main pipeline liquid valve, and injecting the clean seawater stored in the water storage tank into the Venturi gas-liquid mixing and dissolving device through the water pump; the gaseous oxygen active fragments entering the Venturi gas-liquid mixing and dissolving device in the step 2) and seawater are fully mixed and dissolved in the Venturi gas-liquid mixing and dissolving device and the liquid-liquid mixing and dissolving device to generate high-concentration hydroxyl radical solution, and the high-concentration hydroxyl radical solution enters the morbidity culture pond after passing through a TRO detector through a pipeline; the flow rate of the seawater is 10-50L/min, and the concentration of a total oxidant TRO of the hydroxyl radical solution is 2-30 mg/L;
4) after the hydroxyl radical solution is injected into the ill culture pond, the bottom of the ill culture pond is soaked to stimulate cryptocaryon irritans to encapsulate for 0.5-2.0 h, the hydroxyl radical solution oxidizes a weak area of an encapsulating wall to enable the encapsulating wall to generate holes to enter the encapsulating wall, substances forming a basic structure of cells and functional substances generate irreversible reactions, the irreversible reactions comprise amino acid oxidative decomposition and DNA chain breakage, the encapsulating wall is dead, further development and larva release cannot be realized, and residual hydroxyl radicals are decomposed into O2And H2O;An automatic rotating brush controlled by a motor is arranged in the middle of the disease incidence culture pond, and the automatic rotating brush automatically rotates and cleans a bottom capsule after being started; rotating for 10min, and opening an outward liquid discharge valve to discharge dead cysts in the diseased culture pond along with seawater; the cryptocaryon irritans cysts are soaked in a hydroxyl radical solution for 0.5-2.0 h, after the cysts are determined to be dead through a microscope microscopic examination mode, an automatic rotating brush switch is started, and after the cysts are rotated for 10min, the dead cysts in the morbidity culture pond are discharged along with seawater.
2. The method for killing Cryptocaryon irritans cysts by hydroxyl radicals as claimed in claim 1, wherein in step 1), the pathogenic culture pond is a cylinder-like open cement pond with a radius of 2m, a depth of 1.5m and a total volume of 20m3
3. The method for killing and stimulating cryptocaryon irritans by hydroxyl radicals as claimed in claim 1, wherein in step 2), the power of the high-frequency high-voltage power supply is 150-300W, the air input of oxygen is 1.0-5.0L/min, and the oxygen is ionized and dissociated after being bombarded by high-energy electrons to form gaseous oxygen active fragments.
4. The method of claim 3 wherein said gaseous oxygen reactive fragments comprise O2 +、O(1D)、O2(a1g)、O2 -、O(3P) and O3Particles with a concentration of 50-300 mg/L; the pipeline is a polytetrafluoroethylene pipe.
5. The method for killing Cryptocaryon irritans cysts by hydroxyl radicals as claimed in claim 1, wherein in step 3), the water storage tank is a cubic closed cement tank with a total volume of 20m3The stored seawater is clean seawater pretreated by sand filtration.
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