KR102549525B1 - Active Treatment Agent Composition for Simultaneous Control of Olpidiopsis blight and Red-Rot Disease - Google Patents

Abstract

The present invention can effectively control oomycotic gabybyeong, which causes the greatest damage to seaweed farms, to promote the growth of seaweed while preventing the disease of seaweed, and to control oomycotic seaweed disease of seaweed, which can improve the quality of harvested seaweed. It relates to an active treatment agent, and more particularly, to an active treatment composition for simultaneous control of cystomycosis and red foot disease, characterized in that it contains a mixture of lactic acid, propionic acid and hydrochloric acid.

Description

Active Treatment Agent Composition for Simultaneous Control of Olpidiopsis blight and Red-Rot Disease}

The present invention can effectively control cysticercosis and cystitis, which cause the greatest damage to laver farms, at the same time, thereby promoting growth of laver and improving the quality of harvested laver while preventing disease of laver. It relates to an active treatment agent for simultaneous control of.

Seaweeds have been used worldwide for thousands of years as food and dietary supplements, animal feed, chemicals and biofuels. Especially in Asia, where seaweeds are mainly used as food, the aquaculture industry of seaweed, seaweed, and kelp has developed. According to statistics in 2017, among the main aquaculture species such as seaweed, seaweed, and kelp, seaweed accounts for 29.6% of total production and 71% of total production value. is showing

Types of seaweed cultivated in Korea include radiation-patterned seaweed ( Pyropia yezoensis ), it-body stone seaweed ( Pyropia dentata ), and hair-patterned seaweed ( Pyropia seriata ). is a growing trend. The production of seaweed is greatly influenced by natural factors such as seawater, climate and marine conditions, and by seaweed disease, which causes the loss of fronds in seaweed fronds and fading of seaweed fronds. In particular, in the case of seaweed bottles, the quality of seaweed is deteriorated or production is reduced, causing great damage to fishermen. The representative dog diseases in Korea are Olpidiopsis blight, Green-spot disease, and Red-rot disease. Occurs mainly in March.

Among seaweed diseases, cystitis and cystomycosis caused by Pythium porphyrae and Olpidiopsis spp. belonging to the Oomycete River cause the greatest damage to seaweed farms. This is closely related to the characteristics of the reproduction and infection mechanisms of the two disease bacteria, which can continuously form and release zoospores that directly infect seaweed. As a result, if two types of seaweed disease appear at the same time in the seaweed farm, it will cause great damage to the seaweed farm.

In laver farms in Korea, mineral acid, which is 35% hydrochloric acid, has been mainly used as a measure to control seaweed disease. In 1994, the use of acid treatment agents was enacted as a notification, and the use of inorganic acids was restricted. Since 1998, a mixture of organic acids and inorganic acids has been developed and used as an active treatment agent to replace inorganic acids. Currently, all active treatment agents that can be used in seaweed farms contain 8.0 to 9.5% of hydrochloric acid and 10% or more of organic acid. In reality, organic acids are very expensive, so they rarely use more than 10% in practice, and if hydrochloric acid is not used, deposits and residues are not removed, so fishermen do not use them at all, so it is not realistic.

Organic acid, which is the main component of the active treatment agent, is an acidic organic compound that suppresses the growth of mold and microorganisms and is widely used to prevent spoilage of food. The antibacterial effect and range of action of organic acids show differences depending on the type and biochemical characteristics of organic acids. As for the antibacterial effect of organic acids related to dog's disease, the efficacy of lactic acid, malic acid, citric acid, gluconic acid, and tartaric acid has been reported against the bacillus that causes rosacea, but the antibacterial effect of organic acids against cystomycosis has not been reported. In addition, it has been reported that there is no control effect because the host bacillus in the seaweed cells is not damaged even when hydrochloric acid is treated, which was the existing method of controlling ciliary disease.

In addition to this, since cystomycosis and red mud disease almost always occur in seaweed farms, substantial dog disease relief is not achieved unless both diseases are simultaneously caught. In the case of controlling only one disease, there is a high possibility that other diseases will spread easily. As fishermen respond only by handling high concentrations of hydrochloric acid, many illegal activities and various social problems have occurred.

In addition, the active treatment agent developed to replace hydrochloric acid has a higher treatment cost and longer treatment time than hydrochloric acid, and illegal use of hydrochloric acid, an inorganic acid, is still widespread, which has become a social problem.

Registered Patent No. 10-1752610 Patent Publication No. 10-2000-0060843 Japanese Patent Registration No. 3595004 Japanese Patent Registration No. 3631466

The present invention can effectively control cystomycosis and red seaweed at the same time, thereby promoting the growth of seaweed and improving the quality of harvested seaweed while preventing diseases of seaweed. aims to

The present invention for achieving the above object relates to an active treatment agent composition for controlling cystococci, characterized in that it comprises a mixture of propionic acid and hydrochloric acid.

As can be seen in the following examples, hydrochloric acid, which has been conventionally used for the control of oomycete disease, does not show a significant effect on the control of cystomycosis, and the antibacterial effect of organic acids against cystomycosis has not been reported either. However, the mixture of propionic acid and hydrochloric acid of the present invention can very effectively control cysticercosis.

The composition of the present invention also exhibits a synergistic effect on the control of rosacea by mixing propionic acid and hydrochloric acid, so it is effective in controlling not only cysts but also rosacea. It can be used as an active treatment composition for simultaneous control of cysts and rosacea. there is. According to the composition of the present invention, since it is possible to simultaneously control cysticercosis and rosacea, it is possible to substantially rescue oomycete disease.

It is more preferable to further contain lactic acid in order to further amplify the control effect on red mud disease using the composition of the present invention.

According to the permissible content of ingredients in the active treatment agent in Article 4 of the standards for use of active treatment agents for seaweed farms, the total content of chloride ions, sulfate ions, and nitrate ions in active agents mainly composed of organic acids is 8% or more and 9.5% or less. Therefore, the weight ratio of hydrochloric acid in the composition of the present invention is preferably 8% to 9.5%, but the above range is not excluded. As can be seen in the examples, even when containing 5% by weight of HCl, which is a concentration lower than the above range, an excellent control effect was exhibited, and the control effect does not decrease when it is out of the above range. Hydrochloric acid in the composition of the present invention is one of the main components that give the synergistic effect of the composition, and hydrochloric acid itself does not show the control effect of oomycetes at the corresponding concentration, but when mixed with propionic acid, the effect of significantly increasing the control effect is When mixed with lactic acid, it showed a synergistic effect on the control effect against red mud disease.

The concentration of propionic acid in the composition of the present invention is preferably 0.5 to 10% by weight. In addition, in the composition additionally containing lactic acid, the weight ratio of lactic acid to propionic acid is preferably 7:3 to 5:5, which shows similar control effects against cystomycosis and red foot disease. However, the ratio may be appropriately adjusted according to the occurrence pattern of cystomycosis and red foot disease. In other words, as the ratio of lactic acid increases, it is advantageous to control rosacea, and as the ratio of propionic acid increases, it is advantageous to control cystomycosis. The weight ratio of propionic acid can be adjusted appropriately.

The active agent composition of the present invention can control oomycete disease by diluting it with seawater and treating seaweed for 5 seconds to 1 minute. The dilution factor may be 10 to 100 times (v/v), but is not limited thereto and may be appropriately adjusted according to the type or frequency of oomycete disease.

As described above, according to the active treatment composition for the control of oomycete fungus of laver of the present invention, it exhibits a control effect not only against rosacea but also against cystomycosis, which has not been effective in the past, and effectively treats both erythema and cystomycosis even at low concentrations It can prevent damage to seaweed farms and improve the quality of harvested seaweed.

1 is a photomicrograph and culture photo of radiation patterned seaweed.
Figure 2 is a photomicrograph of seaweed lobes infected with rosacea and cystomycosis.
Figure 3 is a graph showing the infection inhibitory efficacy of erysipelas according to the treatment concentration of propionic acid and lactic acid.
Figure 4 is a graph showing the infection inhibitory efficacy of propionic acid or a mixed solution of lactic acid and hydrochloric acid against red dog's disease.
Figure 5 is a graph showing the infection inhibitory efficacy against red mud disease according to the dilution ratio of propionic acid or a mixed solution of lactic acid and hydrochloric acid.
Figure 6 is a graph showing the infection-inhibiting efficacy against red mantis according to the dilution ratio of 9.5% hydrochloric acid aqueous solution.
Figure 7 is a graph showing the infection inhibitory efficacy against red mud disease according to the dilution ratio of lactic acid, propionic acid and hydrochloric acid mixture.
Figure 8 is a graph showing the infection inhibitory efficacy against cystic disease according to the dilution ratio of 9.5% hydrochloric acid aqueous solution.
Figure 9 is a graph showing the infection inhibitory efficacy against cystic disease according to the dilution ratio of the mixed solution of propionic acid and hydrochloric acid.
Figure 10 is a graph showing the infection inhibitory efficacy against cystic disease according to the dilution ratio of the mixed solution of lactic acid and hydrochloric acid.
Figure 11 is a graph showing the infection inhibitory efficacy against cystic disease according to the dilution ratio of a mixture of lactic acid, propionic acid and hydrochloric acid.
Figure 12 is a graph showing the control effect of rosacea and cystic disease according to the mixing ratio of propionic acid and lactic acid.

Hereinafter, the present invention will be described in more detail with reference to the attached examples. However, these embodiments are only examples for easily explaining the content and scope of the technical idea of the present invention, and thereby the technical scope of the present invention is not limited or changed. It will be obvious to those skilled in the art that various modifications and changes are possible within the scope of the technical spirit of the present invention based on these examples.

[Example]

Cultivation of seaweed and strains

1) Radial seaweed ( Pyropia yezonesis )

Radiated seaweed was purchased from the Mokpo Seaweed Bio Research Center and cultured alone in PES (Provaoli Enriched Seawater) medium. Seawater filtered with filter paper (Whatman No. 2) was sterilized with a high-pressure steam sterilizer for 20 minutes under conditions of 121° C. and 1.2 atm. The culture conditions were 10℃, 12:12 hr L:D cycle, white light (>50 μmol/m ² ·s ¹ ), and aeration was performed. The culture medium was replaced every 7 days.

1A is a microscope image (scale bar: 20 μm) of radiation-patterned seaweed, and B is a photograph of culturing radiation-patterned seaweed halves in a PES medium.

2) Red sea blight ( Pythium porphyrae )

The rosacea fungus was isolated and cultured from seaweed lobes infected with erythema collected in Hoedong, Jindo-gun. 2A is a photomicrograph of seaweed lobes infected with R. Unlike normal cells in FIG. 1, traces of hyphae passing through Kim cells like threads can be seen in A of FIG. The infected area of the collected seaweed was cut and subcultured until separated from the cornmeal agar medium. The isolated strain was subcultured in a cornmeal agar medium at 20°C and a 12:12 hr L:D cycle for 21 days. When used in the experiment, the separated mycelia were used by suspension culture in Arasaki B medium for 7 to 10 days under the same light and temperature as the subculture conditions.

3) Mycobacterium ( Olpidiopsis pyropiae )

Cystic bacillus was isolated and cultured by transferring the disease to radiation-patterned seaweed from seaweed lobes infected with cystomycosis collected from Hoedong, Jindo-gun. Figure 2B is a photomicrograph of seaweed fronds infected with cystomycosis, showing that the seaweed cells turned yellow due to infection with cystomycosis. The culture conditions were 15 ℃, 12:12 hr L:D cycle, white light (>50 μmol / m ² · s ¹ ) was used, and new radiation was added every 5 to 7 days to maintain cyst bacteria.

Induction of cyst infection and measurement of infection rate

1) Red mud bottle

The mycelial rosacea fungus, which was cultured in suspension for 7 to 10 days in Arasaki B medium, was exposed to sterilized seawater for 24 to 30 hr to induce the release of zoospores. The cell number of released zoospores was measured using a hemocytometer after fixation in a fixative diluted with 3.7% formaldehyde at a rate of 1/500. Zoospores of 2.5 × 10 ³ cell / ml were infected by culturing in sterilized seawater with radiation. The infection was observed and photographed using an optical microscope (OLYMPUS, BX50) and cellSens program. The infection rate was measured by comparing the infected area to the total area of the photograph taken using the image program Adobe Photoshop.

In the following examples, the statistical difference in the infection rate and the number of infected cells according to the type of treatment solution and the dilution rate was confirmed through one-way ANOVA. The difference according to the type and dilution ratio of each treatment solution was confirmed through a post-hoc test.

2) Cystomycosis

After releasing zoospores from seaweed fronds infected with cystomycosis, the infected water containing zoospores was filtered through a 40 μm filter to infect radiation. Observation and filming of infection were conducted in the same way as for red mantis.

Efficacy test of active treatment agent for red mud disease

1) Infection analysis according to concentrations of propionic acid and lactic acid treatment

In order to analyze the minimum effective concentrations of propionic acid and lactic acid, which have excellent inhibitory effects on red mantis infection, samples by concentration (0.1, 0.25, 0.5, 1 wt%) were prepared, and then treated for 15 seconds on the thallus infected with red mantis. Immediately after treatment, the thallus was washed in sterilized seawater and cultured for 3 days to confirm that the infection of the control group had progressed by more than 80%, and then the infection rate of the treatment group was compared and measured.

Figure 3 shows the results. Propionic acid (PPA) was not significantly different from the infection rate of the control group until 0.25% treatment, and the infection rate (52.7%) of the 0.5% treatment group was lower than that of the control group (90.9%), The lowest infection rate (5.63%) was shown in the 1% treatment group. In addition, it can be seen from FIG. 3 that lactic acid (LA) exhibits an effect at a lower concentration than propionic acid. The infection rate of 84.5% in the 0.1% treatment group was higher than the infection rate of 4.0% in the 0.25% treatment group, 4.2% in the 0.5% treatment group, and 3.9% in the 1% treatment group, but significantly lower than the infection rate of 90.9% in the control group.

The pH of propionic acid was 2.99 in 1% solution and 3.83 in 0.1% solution, and the pH of lactic acid was 2.24 in 1% solution and 3.02 in 0.1% solution, indicating that lactic acid was more acidic than propionic acid.

2) Infection analysis according to treatment of mixed solution of organic acid and hydrochloric acid

According to the standards for use of activator for seaweed farms, the allowable content of each component in an activator containing organic acid as the main component is 10% or more for organic acid, 8% or more and 9.5% or less for the total content of chlorine ion, sulfate ion, and nitrate ion, sulfuric acid ions and nitrate ions are each less than 2%.

Accordingly, an activating agent having a composition of propionic acid (10 w%) + hydrochloric acid (9 w% or 5 w%) and lactic acid (10 w%) + hydrochloric acid (9 w% or 5 w%) was prepared. After diluting the active treatment agent with 1/30 sterilized seawater, it was treated for 15 seconds on the fronds infected with red mantis, and then immediately washed with sterilized seawater. After culturing for 3 days, the infection rate of the treatment group was compared and measured when the infection of the control group had progressed to 80% or more.

The infection rate of the control group was 83.7%, compared to the 10% propionic acid treatment group's infection rate of 61.9%, the 5% hydrochloric acid treatment group's infection rate of 69.2%, and the 9.5% hydrochloric acid treatment group's infection rate of 65.3%. The infection rate was 3.4% and 3.0%, respectively, and the effect was very remarkable when treated with a mixture of propionic acid and hydrochloric acid.

On the other hand, since the infection rate of the 10% lactic acid treatment group was already very low at 1.9%, even when additional 5% hydrochloric acid or 9.5% hydrochloric acid was mixed, there was no significant difference compared to the lactic acid only treatment.

3) Infection analysis according to the dilution ratio of the mixture of organic acid and hydrochloric acid

Infection was analyzed by the same method as in 2), except that the mixed solution of organic acid and hydrochloric acid was diluted.

The infection rate of the 10% propionic acid and 9.5% hydrochloric acid mixture increased as the dilution ratio increased, and the infection rate of the 80-fold dilution was similar to that of the control group. For reference, when only 9.5% hydrochloric acid was diluted and treated, there was no significant difference between the control group and the infection rate (see FIG. 6). The infection rate of the 50-fold diluted propionic acid treatment group was 21.3%, which was significantly reduced to 25% of the control group's infection rate of 86.1%. The concentration of propionic acid in the solution was 0.2%, suggesting that the mixed solution had a synergistic effect because neither the propionic acid solution (FIG. 3) nor the hydrochloric acid solution (FIG. 6) at that concentration showed the control effect of erythema.

The mixture of 10% lactic acid and 9.5% hydrochloric acid showed a tendency to increase the infection rate as the dilution ratio increased, but there was no significant difference. The infection rates of the 50-fold, 60-fold, and 80-fold dilutions were 3.3, 4.2, and 5.2%, respectively, which were about 3.8 to 6.0% compared to the control group's infection rate of 86.1%. Since lactic acid itself has a very large infection-inhibiting effect, the synergistic effect by mixing with hydrochloric acid was weaker than that of propionic acid, but when the concentration was converted with reference to FIGS. 3 and 6, could confirm that

4) Infection analysis according to the treatment of lactic acid, propionic acid and hydrochloric acid mixed solution

After preparing an active inhibitor by mixing lactic acid, propionic acid, and hydrochloric acid to the concentrations shown in Table 1, the degree of inhibition of rosacea infection was evaluated by the same method as in 2), except that the diluted solution was treated. The results are shown in Table 1 and FIG. 7 . In FIG. 7, A, B, and C represent experimental groups 1, 2, and 3 in Table 1, respectively.

The pH of the solution according to the dilution ratio of 9.5% hydrochloric acid and the composition of Table 1 was measured with a pH meter (NeoMet, pH/ISE Meter pH-250L) attached to a glass electrode (Hemilton, Liq-Glass BNC), and the results are shown in the table. 2.

Efficacy test of active treatment agent for cystomycosis

1) Infection analysis according to the dilution ratio of the mixture of organic acid and hydrochloric acid

A mixture of organic acid and hydrochloric acid was diluted according to the same method as for red midge disease, and treated with seaweed fronds infected with cystomycosis, and the number of infected cells was observed.

First, FIG. 8 is a graph showing the infection rate of cysts according to the dilution ratio of 9.5% hydrochloric acid, and the infection rate decreased by about 20% according to the treatment of 9.5% hydrochloric acid dilution, but it was not significant.

Figure 9 is a graph showing the infection rate of Cyst bacilli according to the dilution treatment of a mixture of 10% propionic acid and 9.5% hydrochloric acid. The number of infected cells in the mixture of propionic acid and hydrochloric acid diluted 30, 40, and 50 times was less than that of the control group (98.6). There was no significant difference between the number of infected cells in the 30-fold dilution treatment group (6.6 cells) and the 40-fold dilution treatment group (10.5 cells), and the number of infected cells in the 50-fold dilution treatment group (30.2 cells) was less.

The mixture of 10% lactic acid and 9.5% hydrochloric acid showed a tendency to increase the infection rate as the dilution ratio increased. The number of infected cells when treated with 30-fold, 40-fold, and 50-fold dilutions was 60.6, 73.8, and 89.8, respectively, which was 61-91% compared to 98.6 infected cells in the control group. The number of infected cells at 50-fold dilution was not significantly different from that of the control group. A dilution containing lactic acid showed a strong effect against red mantis, but a dilution of lactic acid was less effective than propionic acid against Cyst bacilli.

2) Infection analysis according to the treatment of lactic acid, propionic acid and hydrochloric acid mixed solution

After preparing an active inhibitor by mixing lactic acid, propionic acid, and hydrochloric acid to the concentrations shown in Table 3, the degree of inhibition of Cystomycosis infection was evaluated by the same method as in 1), except that the diluted solution was treated. The results are shown in Table 3 and FIG. 11.

Confirmation of the control effect of erythrombosis and cystomycosis according to the concentration of the composition for controlling oomycete gallbladder disease

In order to confirm the optimal composition of the composition for the simultaneous control of red mud disease and cystomycosis, 9.5% hydrochloric acid and a mixture of lactic acid and propionic acid were compared with the control effect while changing the ratio of lactic acid and propionic acid.

12 is a graph showing the treatment results using a 30-fold dilution, where LA represents lactic acid and CP represents propionic acid. Figure 12 shows that when the weight ratio of propionic acid to lactic acid is 7:5 to 3:5, it is possible to control rosacea and cystomycosis at the same time with excellent efficiency.

Claims (9)

delete delete delete delete An active treatment composition for simultaneous control of cystomycosis and red foot disease, comprising 0.5 to 10% by weight of propionic acid and 8 to 9.5% by weight of hydrochloric acid.
The method of claim 5,
An active treatment composition for simultaneous control of cystomycosis and rosacea, characterized in that it additionally contains lactic acid so that the weight ratio of lactic acid and propionic acid is 7: 3 to 5: 5.
delete According to claim 5 or claim 6,
An active treatment composition for simultaneous control of cystomycosis and red foot disease, characterized in that the active treatment composition is diluted with seawater and treated for 5 seconds to 1 minute. delete

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