JP6367623B2 - Minamata protozoan pesticide and method for producing the same - Google Patents

Description

  The present invention relates to a pesticide for controlling aquatic protozoa parasitizing freshwater appreciating fish such as goldfish and carp, and a method for producing the same.

  Freshwater saltwater fish such as goldfish and brocade fish are caused by parasitic parasites such as Dactylogyrus vastator, Gyrodactylus elegans, Costia necatrix, Chilodonella cyprini, and Ichityophthirius sp. It is known to cause various diseases. In order to control these harmful varicella protozoa, commercially available anthelmintic agents mainly composed of trichlorfone are used. However, these commercially available anthelmintic agents have medicinal effects on, for example, daktiro, kirodak, and kilodonella. To expect, it is necessary to continue the treatment for at least 5 days while controlling the temperature of the aquarium. Moreover, even when such measures are taken, the survival rate of Minamata protozoa is as high as 40%, and it is difficult to obtain the expected effect.

  In addition, since these commercially available anthelmintic agents have little effect on ostia protozoa, in particular, when they are targeted, it is necessary to carry out a medicine bath using formalin. The residual rate is 20% or more, and it is difficult to obtain the expected effect. Moreover, formalin is toxic, and if wastewater containing it is allowed to flow as it is, there is a risk of adverse effects on the environment.

  Instead of such a conventional anthelmintic method, a method mainly using a tea extract has been proposed. For example, Patent Document 1 below discloses that the use of an extract of green tea such as catechins can prevent or treat bacterial fish diseases such as nodular diseases, Jacolo disease, Aeromonas disease, and gypsosis. Has been. Moreover, the following patent document 2 discloses that a tea extract can control an ichthyodo parasite that parasitizes salmon and trout.

JP-A-4-103537 JP 2006-219478 A

  By the way, hot water treatment is generally used to extract catechins from tea leaves as described above, but the extracted hot water extract contains caffeine that is toxic to fish in addition to catechins having medicinal properties. Many are included. Therefore, it is necessary to completely remove toxic caffeine and extract only catechins. However, this requires a lot of labor and increases costs.

  In addition, there is a method using commercially available powdered catechins, but these are generally expensive, resulting in high costs for farmers. In addition, commercially available powdered catechins need to be added in a large amount in order to obtain a sufficient effect, and an antiparasitic agent that exhibits an excellent effect when added in a small amount is desired.

  Therefore, the present invention has been devised to solve these problems, and an object of the present invention is to provide a novel varicella protozoan pesticide capable of exhibiting excellent medicinal effects at a low production cost and a method for producing the same. Is.

  In order to solve the above-mentioned problems, a first invention is a protozoan protozoan control agent comprising catechins and tannin, wherein the tannin is a complex of catechins and caffeine. According to such a structure, the anthelmintic effect superior to the case of using a catechin simple substance can be exhibited with respect to the protozoan parasite parasitizing freshwater ornamental fish, such as a goldfish and a Nishikigoi. In addition, since caffeine that is toxic to fish is inactivated by being complexed with catechins, it does not harm fish.

  A second invention is the varicella protozoan pesticide which further comprises saponin in the first invention. Saponins have a hydrophilic group and a hydrophobic group, and are known to have a surface-active effect. Therefore, the effect that the protozoan protozoa attached to the fish body by this saponin is easily peeled off from the fish body surface can be exhibited.

A third invention is the varicella protozoan control agent characterized in that in the first or second invention, (-) epigallocatechin gallate is most contained among the catechins. According to such a structure, since the (-) epigallocatechin gallate which shows the especially outstanding medicinal effect among several catechins is contained in high concentration, the outstanding anthelmintic effect can be exhibited even with a small quantity.

  A fourth invention is the varicella protozoan pesticide according to the first to third inventions, wherein the amount of the caffeine is smaller than the total amount of catechins in the tannin. According to such a configuration, since most of the contained caffeine is deactivated as a complex with catechins, it is possible to suppress the adverse effect of caffeine on fish.

  According to a fifth invention, in any one of the first to fourth inventions, the amount of the tannin is substantially equivalent to the sum of the catechins and caffeine contained in the tannin. It is a pesticide. According to such a configuration, since most of the contained caffeine is deactivated as a complex with catechins, it is possible to suppress the adverse effect of caffeine on fish. Moreover, since it is possible to save time and effort to remove caffeine more than necessary, the manufacturing cost is also reduced.

  The sixth invention is a boiling / washing process in which tea leaves are boiled for a predetermined time to wash the tea leaves after boiling, and the tea leaves washed in the boiling / washing process are boiled again for a longer time, and then the tea leaves are separated. Tea leaves / boiling liquid separating step for recovering boiling liquid A and tea leaves separated in the tea leaf / boiling liquid separating step are pulverized while boiling again, and pulverized tea leaves are separated to recover boiling liquid B. A cream that causes a cream-down phenomenon by mixing the boiling liquid B obtained in the liquid recovery step, the crushing / boiling liquid recovery step and the boiling liquid A obtained in the tea leaf / boiling liquid separation step. A method for producing a varicella protozoan pesticide, comprising a down step. According to such a production method, the varicella protozoan pesticide as in the first to fourth inventions can be produced easily and at low cost. Each step will be described in detail later.

  According to a seventh aspect of the present invention, in the sixth aspect, the pulverization step is characterized in that the tea leaves separated in the separation step are boiled again in a pressure cooker and then rapidly decompressed to crush the tea leaves. It is a manufacturing method of an agent. According to such a production method, tea leaves can be easily pulverized and (−) epigallocatechin gallate having particularly excellent medicinal properties can be efficiently extracted therefrom. This step will also be described in detail later.

  According to the present invention, an anthelmintic effect superior to the case where catechin is used alone can be exerted against varicella protozoa parasitizing freshwater ornamental fish such as goldfish and Nishikigoi. In addition, caffeine that is toxic to fish is inactivated by forming a complex with catechins, so that it does not harm fish. Furthermore, since it is not necessary to completely remove caffeine, it can be produced at low cost.

It is a schematic diagram which shows the component structure of the varicella protozoan pesticide 100 which concerns on this invention. It is a table | surface figure which shows the component test result of the varicella protozoan pesticide 100 which concerns on this invention. It is a flowchart figure which shows the manufacturing method of the varicella protozoan pesticide 100 which concerns on this invention. It is a graph which shows the relationship between the boiling time of a tea leaf, and the extraction rate of each containing component. It is a graph which shows the relationship between the survival rate of goldfish at the time of raising only with clear water, and elapsed time. It is a graph which shows the relationship between the survival rate of goldfish at the time of adding the varicella protozoan pesticide 100 which concerns on this invention, and elapsed time. It is a graph which shows the relationship between the survival rate of goldfish at the time of adding commercially available powdery catechin, and elapsed time.

Next, embodiments of the present invention will be described with reference to the accompanying drawings. FIG. 1 shows the component constitution of the varicella protozoan pesticide 100 according to the present invention, FIG. 2 shows the test results, and FIG. 3 shows the flow of the manufacturing method. As shown in FIG. 1, aquatic protozoa pesticides 100 according to the present invention is an agent of the liquid composed mainly of tea hot water extract, other polyphenol 10 and saponin 40 of water (H 2 _O) Is included. Furthermore, the polyphenol 10 is mainly composed of catechins 20 and tannin 30.

  As shown in the figure, the catechins 20 are mainly (−) gallocatechin, (−) epigallocatechin, (+) catechin, (−) epicatechin, (−) epigallocatechin gallate, (−) gallocatechin gallate. , (−) Epicatechin gallate, (−) catechin gallate, and pravonoids.

  The tannin 30 is composed of the above-described complex of catechins, that is, a complex of catechins and caffeine, and many of them are contained as white floating matters or precipitates. Saponin 40 does not form a complex with catechins and exists alone.

  And as content of each of these components, as shown in FIG. 2, in each 100 ml, (-) gallocatechin: 31.2 mg, (-) epigallocatechin: 24.3 mg, (+) catechin: 4.5 mg , (−) Epicatechin 6.7 mg, (−) epigallocatechin gallate: 43.8 mg, (−) gallocatechin gallate: 26.2 mg, (−) epicatechin gallate: 10.1 mg, (−) catechin gallate: It is 4.2 mg (May 15, 2014, researched by EcoPro Research Co., Ltd.). In particular, the varicella protozoan control agent 100 according to the present invention is characterized in that it contains the largest amount of (−) epigallocatechin gallate having the highest anthelmintic effect among these eight types of catechins.

  In addition, as shown in the figure, the content of caffeine is 10.8 mg, but these are the amounts contained in the tannin as a complex, and exist as a simple substance (isolated). Absent. Moreover, content of the tannin which exhibits the excellent anthelmintic effect similarly to catechins is 0.16g (160mg), and is the largest among each component. Saponin is a by-product when tannin is formed, and its content is 45.0 mg.

  Next, an example of a method for producing the varicella protozoan pesticide 100 of the present invention comprising such components will be described with reference to the flowchart of FIG. In order to produce the pesticide 100 of the present invention, first, tea leaves are boiled for a short time and caffeine is extracted as the first step 1. Specifically, prepare 2 liters of hot water (boiling water) in a pot, put about 150 g of tea leaves into it, boil it for about 3 and a half minutes (about 210 seconds), then boil it as the next step 2 Drain the dried tea leaves with a colander and wash with running water (tap water) for at least 5 minutes.

  That is, caffeine contained in tea leaves is extracted in the first step 1, and the caffeine adhering to the tea leaves is removed by washing the tea leaves with running water in the next step 2. FIG. 4 shows the relationship between the boiling time (extraction time) of tea leaves and the extraction rate of each component. When tea leaves are boiled with hot water, caffeine is first extracted, and then catechins are extracted. In step 1, boiling is stopped when more than half of caffeine is extracted (about 3 and a half minutes). This makes it possible to remove more than half of caffeine from tea leaves and at the same time suppress the outflow of catechins from tea leaves.

  In the next step 3, the tea leaves washed in step 2 are again boiled with hot water for a long time. Specifically, prepare 4 liters of hot water (boiling water) in the pan, put the washed tea leaves with well drained water and boil them for 30 minutes, and then in step 4 the tea leaves and boiling liquid Separate A. The boiled liquid A thus obtained has a small amount of caffeine, and instead contains a large amount of catechins.

  In the next step 5, the tea leaves separated in step 4 are used again and boiled at a high temperature and high pressure (for example, 2 atm, 120 ° C.). Specifically, 2 liters of hot water (boiling water) is put into a pressure cooker, the tea leaves separated in step 4 are put therein, and after boiling for about 10 minutes with the lid of the pressure cooker closed, The pressure cooker is depressurized at once by opening the pressure cooker's pressure valve. By this treatment, the boiling liquid boils vigorously inside the pressure cooker, and the tea leaves are crushed apart by the momentum, and (−) epigallocatechin gallate, which is particularly difficult to extract, is extracted at once from the cells of the tea leaves. Will be.

  In the next step 6, when the boiling is finished, the contents in the pressure cooker are poured into a fine colander (filter), and the tea leaves that are crushed apart, and (-) the epigallocatechin gallate is extracted. Liquid B is separated and the tea leaves are discarded as they are. On the other hand, after the boiling liquid B is mixed with the boiling liquid A previously separated in the step 5 in the next step 7, the mixed solution C is subjected to a cream down process in the next step 8. Cream-down is a phenomenon in which caffeine and catechins react to form tannin and crystallize.

  Specifically, the container containing the mixed solution C is covered with a heat insulating agent such as a towel so that the mixed solution C at a high temperature (80 ° C. or higher) is cooled more slowly than when it is naturally cooled. Alternatively, the cream-down phenomenon can be reliably generated by immersing the container in hot water (bath) of about 40 ° C. In addition, saponin is also produced | generated simultaneously as a by-product of the tannin produced | generated by this cream down phenomenon. If a tannin suspension or precipitate is produced by this cream-down treatment, the tannin is then stored in a refrigerator to prevent decomposition.

  The thus obtained varicella protozoan pesticide 100 of the present invention is appropriately added in an amount of about 0.00001 to 0.0010 wt% (0.1 to 10 ppm) in terms of tannin in a water tank or a ginger containing a target fish. Therefore, since not only the catechins contained but also tannin exerts an anthelmintic effect against protozoan protozoa, it is possible to exert a superior medicinal effect as compared with the case where only catechins are used. That is, tannins produced for the purpose of deactivating caffeine that is toxic to fish act synergistically with catechins, and thus can exhibit excellent medicinal effects that cannot be obtained with catechin alone.

  Furthermore, the varicella protozoa control agent 100 of the present invention contains a larger amount of (−) epigallocatechin gallate, which is particularly superior in catechins, so that even catechins alone are more effective than ordinary tea extracts. Can be demonstrated. In addition, caffeine that is toxic to fish alone is inactivated by forming a complex with catechins, so it does not cause any harm to fish. Therefore, a process for completely removing caffeine is not required, and no special chemicals or equipment such as alcohol and organic solvent are required for the production of the product of the present invention.

  That is, the present invention does not completely remove caffeine that is toxic to fish, but inactivates it using the cream-down phenomenon, and its product tannin has a synergistic effect together with catechins. It is characterized by the fact that it can be demonstrated, and the fact that it has a high anthelmintic effect but is difficult to extract by simple boiling treatment (-) epigallocatechin gallate was extracted efficiently by crushing tea leaves using pressure fluctuation etc. is there. Based on these technical ideas, the protozoan protozoa repellent 100 of the present invention is low in production cost and has an excellent medicinal effect (antiparasitic effect) against varicella protozoa such as dactyro, kildaku, costair, kilodonella, and white spot protozoa. It can be demonstrated.

  In addition, the saponin produced together with tannin in this cream down treatment has a hydrophilic group and a hydrophobic group and has a surface-active effect, so that the protozoa attached to the fish surface by this saponin is easily peeled off from the fish surface. There is also. Some saponins have fish toxicity, but the saponins produced by the cream-down treatment according to the present invention have almost no fish toxicity.

  The type and form of tea leaves that can be used for the raw material of the present invention are not particularly limited, and any one of fresh leaves, gyokuro, sencha, kazuki tea, bancha, hojicha, black tea, etc. or Two or more may be mixed. In addition, since there is little relation between the anthelmintic effect and the production area or grade of tea leaves, the production cost can be reduced by using the cheapest one. In this embodiment, in order to extract and remove caffeine first, tea leaves are put into hot water and boiled and washed for a short time. For example, restaurants, hotels, soft drinks (tea It is also possible to use the used (extracted caffeine) tea leaves as waste from the factory of) in the second and subsequent steps.

Next, examples of the varicella protozoan pesticide 100 of the present invention will be described.
(Example)
Six groups of 300 goldfish artificially infected with daktiro, kildaku, costair, and kilodonella after six days (48 hours) each with six groups (capacity of 54L in each tank, average water temperature of 18 ° C, filtration is aquaculture flower) The first group is not treated at all (Shimizu only), and the second to fifth groups are 0.00001% by weight (0.1 ppm) of the varicella protozoan insecticide 100 of the present invention in terms of tannin, Added in amounts of 0.0001% (1ppm), 0.0003% (3ppm), 0.001% (10ppm) and every 24 hours (24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 148 hours, 172 hours) 196 hours), the number of goldfish moribund (survival rate) for each group was measured over a total of 8 days.

  As a result, as shown in FIG. 5, in the group raised only with fresh water, goldfish began to drown several hours after the start of the test, and the number of drowning increased after 72 hours, and more than half of them drowned after 120 hours. After 192 hours (8 days), all goldfish were drowned.

  On the other hand, as shown in FIG. 6, in the group to which the pesticide of the present invention was added at 0.00001% by weight (0.1 ppm), most goldfish survived until 96 hours, and the number of deaths gradually increased after 96 hours. However, even after 192 hours (8 days) at the end of the test, more than 50% of goldfish survived. Further, in the group to which 0.001% by weight (1 ppm) of the pesticide of the present invention was added, 80% or more of goldfish survived even after 192 hours (8 days) at the end of the test. The survival rate increased as the amount of pesticide added increased, and in the group added with 0.001% by weight (10 ppm), more than 95% of goldfish survived after 192 hours (8 days) at the end of the test. Was.

(Comparative example)
In the same manner as in the Examples, in each aquarium containing 60 goldfish artificially polymerized and infected with dactyro, kirodak, costa, and kilodonella, powdered catechins manufactured by Taiyo Kagaku Co., Ltd. (trade name: Sunphenon) DK) is added 30 times the amount of the product of the present invention, that is, 0.0003 wt% (3 ppm), 0.003% wt (30 ppm), 0.009 wt% (90 ppm), and 0.03% wt (300 ppm) for 24 hours. Each time, the number of goldfish moribund (survival rate) was measured over a total of 8 days.

  As a result, the survival rate of goldfish was higher by adding powdered catechins as shown in FIG. 7 than when not added (only with fresh water), but each was 30 times the amount of the present invention (weight%). In all cases, the survival rate was lower than that of the present invention.

(Discussion)
As shown in the Examples and Comparative Examples, by using the varicella protozoa control agent 100 according to the present invention, the survival rate of freshwater ornamental fish parasitized with varicella protozoa such as dactyro, kirodaku, costair, and kilodonella greatly increases, The effect was higher than when commercially available powdered catechin was used. In particular, when compared with the untreated group, a remarkable survival rate was observed even with the addition of only 0.00001% by weight (0.1 ppm).

  In addition, when the amount of addition was 0.00001% by weight (0.1ppm), 0.0001% by weight (1ppm), 0.0003% by weight (3ppm), and 0.001% by weight (10ppm), as shown in FIG. In the case of 0.00001% by weight (0.1ppm), the survival curve suddenly dropped, but the concentration was high when 0.0001% by weight (1ppm), 0.0003% by weight (3ppm) and 0.001% by weight (10ppm) were compared. Although the effect (survival rate) becomes higher, the difference was not great. Rather, 0.0001% by weight (1ppm) and 0.001% by weight (10ppm) cost 10 times, but the effect is not so different, so in reality it is 0.0001% by weight (1ppm) to 0.0003% by weight (3ppm). It has been found that use in the range is most cost-effective.

  On the other hand, even when powdered catechin was used, the survival rate was improved. However, even if the amount added was 30 times that of the present pesticide, the effect was lower than that of the present pesticide. It was. In this way, the pesticide of the present invention exerted an excellent anthelmintic effect in a very small amount, especially among catechins (-) epigallocatechin gallate is contained in a high ratio, In addition, tannins and saponins generated by the cream-down phenomenon for the purpose of deactivating caffeine act synergistically with catechins, and so on, which is a remarkable anthelmintic as many as 30 times more than when used with conventional catechins alone. It is presumed that the effect was demonstrated. Incidentally, when the amount of commercially available powdered catechin was reduced to the same level as that of the present invention (0.0001 wt% (1 ppm) to 0.001 wt% (10 ppm)), almost no anthelmintic effect was observed.

  Moreover, the commercially available powdered catechin has an average market price of about 2000 yen per 5 g at the time of filing of the present application. For this reason, if this commercially available powdered catechin is added to a 10 L water tank by 0.005% by weight (50 ppm), the cost is about 20,000 yen, and high cost is a problem. On the other hand, in the case of the present invention product, 0.0001% by weight (1 ppm) may be added to obtain the same effect, and the cost is calculated to be only several tens of yen to several hundreds of yen. Compared with commercially available products, significant cost reduction (1/1000 to 1/100) is possible. Further, if used tea leaves from restaurants and hotels, ie, so-called tea leaves, are used as raw materials, the manufacturing cost can be further greatly reduced.

DESCRIPTION OF SYMBOLS 100 ... Minamata protozoan pesticide 10 ... Polyphenol 20 ... Catechin 30 ... Tannin 40 ... Saponin

Claims (2)

A caffeine extraction process in which tea leaves are boiled to extract caffeine;
A washing step of removing the adhering caffeine by washing the tea leaves after extracting caffeine in the caffeine extraction step ;
A catechin extraction step of extracting the catechin by boiling again the tea leaves washed in the washing step ;
A boiling liquid A recovery step of separating the tea leaves from the boiling liquid of the catechin extraction step and recovering the boiling liquid A containing catechin ;
The tea leaves separated in the boiling liquid A recovery step are pulverized while boiling again, the boiling tea liquid B recovery step of separating the ground tea leaves and recovering the boiling liquid B ,
The boiling liquid B obtained in the boiling liquid B collecting step and the boiling liquid A obtained in the boiling liquid A collecting step are mixed, and the mixed liquid C is cooled more slowly than the natural cooling state, thereby causing a cream-down phenomenon. A method for producing a protozoan protozoan pesticide, comprising: In the method for producing a varicella protozoan pesticide according to claim 1 ,
In the boiling liquid B recovery step, the tea leaves separated in the boiling liquid A recovery step are boiled again in a pressure cooker, and then rapidly decompressed to pulverize the tea leaves.

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