JP2011084524A - Algae proliferation inhibitor containing wood component - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an algae proliferation inhibitor containing a wood component and friendly to the environment. <P>SOLUTION: Parts such as leaf, bark, sapwood and heartwood of trees such as cedar, Japanese black pine, Japanese red pine, cypress, Thujopsis dolabrata var, hondae, Cinnamomum camphora, camphor tree, bald cypress and dawn redwood, essential oil obtained by steam distillation or hot-water distillation of the tree parts, effluent containing the essential oil component and an extract of the parts extracted with a solvent such as hexane, ethyl acetate and methanol can be used as the algae proliferation inhibitor effective for preventing red tide, etc., and friendly to the environment. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to an algal growth inhibitor containing a tree component.

Trees have the largest carbon dioxide absorption by terrestrial organisms and are known as one of the renewable resources. Maintaining and developing the forestry / forestry industry by using trees and planting trees also leads to the reduction of carbon dioxide and is essential for the sustainable development of mankind.
However, the amount of bark as a forest waste generated during the lumbering of trees, waste materials, and board materials and pillar materials as building demolition waste are enormous (Non-patent Document 1 and Non-Patent Document 2). Woody biomass accounts for about 60% of the total available amount of resources that are not effectively used in Japan (available amount), and its annual generation amount is 45 million tons (Non-patent Document 3). In recent years, regulations on incineration facilities have been strengthened since December 2002, and the control of carbon dioxide emissions and the toxicity of dioxins have become problems. Yes.

On the other hand, since the mid-1960s in the coastal areas of our country, phenomena such as red tide or shellfish is now occur very ^frequently, a wide area of red tide occurrence waters, the increase in the number of occurrences, the tendency of prolonged generation period seen It has become like this. As can be seen from the occurrence of red tide in the past 10 years up to 2006, the red tide problem has occurred a lot, although it varies from year to year (Fig. 1, Non-Patent Document 4).

As countermeasures for red tide, physical methods (red tide recovery, ultrasound, etc.), chemical methods (accoril, clay injection, etc.) and biological methods (algicidal algae, algaicidal virus, etc.) have been proposed in the past. However, it has not been put into practical use due to problems such as “scale, cost, and safety”. Many damages to the fishery and aquaculture industry due to red tide and shellfish poison are still occurring, and a great deal of damage to the fishery and aquaculture industry has been reported in various sea areas in Japan (Non-patent Document 5). Therefore, the promotion of healthy aquaculture is one of the most important and specific food measures, and the promotion of effective measures against red tide and shellfish poison is an important issue.

As a countermeasure for red tide, physical methods such as collecting red tide on the surface of the sea, circulating seawater to lower the concentration of red tide, and chemical methods such as using clay and its components, spraying acrinol ⁾ , using magnesium hydroxide, etc. In addition, various control methods such as biological methods using algaecidal bacteria and algaecidal viruses that apply the relationship between microalgae and heterotrophic bacteria that affect each other in the ocean have been proposed. However, none has been put into practical use.

Patent Document 1 discloses an algae growth inhibitor containing isoflavones as active ingredients, but the raw material can be food such as soybean and is suitable for an era when the global food crisis is a concern. I can't say that. Patent Document 2 discloses a method in which a chelating agent is dispersed in water, but it is not necessarily an environmentally friendly substance, and a secondary problem may occur.

JP-A-2005-248161 JP 2009-66549 A

Bunichiro Tomita (1999) R & D Tour Seminar “Toward the Construction of Zero-Emission Wood Industry” sponsored by the 34th Nagoya International Woodworking Machinery Exhibition Executive Committee, Japan Wood Society, Nagoya, pp. 1-14. Tsutomu Takano, Katsumi Nishimura, Koji Murata (1999) Waste disposal and usage trends in sawmills, woodworking machines 183, 5-8. Shiro Saka, Katsunobu Ehara, Eiji Minami (2005) Utilization of woody biomass by supercritical fluid technology, Journal of the Wood Society 51, 207-217. Japan Coast Guard (2008) Current Status of Marine Pollution (January-December 2006), Trends in Number of Confirmed Occurrence of Substances by Marine Pollution (for the Past 10 Years) Exercise information version, 2007: www. gyosai. or. jp / topics_pdf / topics63. pdf, more than 200,000 farmed fish dead due to red tide damage in Ehime Prefecture and Uwa Sea

Development of an environmentally friendly algal growth inhibitor is desired.

In addition, there are almost no studies on red tide control methods derived from natural ingredients, and it is not known whether trees can be used as biological control methods for red tides.

In order to identify components that are specifically effective in suppressing the growth of the red tide algae Skeletonema costatum, 10 monoterpenes (10 carbon atoms), 5 sesquiterpenes (15 carbon atoms), diterpenes (20 carbon atoms) 11 types, norlignans 5 types, flavonols 3 types, lignin degradation products, lignans, quinones, tropolones, phenylpropanoids, 1 type each, 10 types and 39 types of tree component preparations and cedar, cypress, As a result of experiments on the suppression of growth of red tide algae Skeletonema costatum and Heterosigma akashiwo using essential oil collected by hot water distillation from the bark, timber and leaves of black pine, both cedar bark and black pine heartwood essential oil are strongly suppressed. Showed the effect. In addition, as a result of analyzing the components of cedar bark and black pine heartwood and examining the activity of the components alone, it was discovered that Ferguinol, a cedar bark component, and longifolene, a pine heartwood component, have particularly high growth inhibitory activity Reached.

There is no report on the red tide algae growth inhibitory activity of tree components such as cedar such as ferguinol and longifolene, cypress and black pine shown in the present invention. In addition, since the cedar bark and black pine heartwood used in the present invention are currently waste and are waste resources for which development of usage is desired, the usefulness and needs of the present invention are great.

The time-dependent change of the number of red tide occurrence in the past 10 years is shown (nonpatent literature 4). A hot water distillation apparatus is shown. The growth inhibitory effect (final average growth rate) with respect to Skeletonema costatum and Heterosigma akashiwo of a tree extract is shown. The growth inhibitory effect (final average growth rate) with respect to Skeletonema costatum of monoterpenes is shown. The growth inhibitory effect (final average growth rate) with respect to Skeletonema costatum of sesquiterpenes is shown. The growth inhibitory effect (final average growth rate) with respect to Skeletonema costatum of diterpenes is shown. The growth inhibitory effect (final average growth rate) with respect to Skeletonema costatum of norlignans, flavonols, lignin degradation products, lignans, quinones, tropolones, and phenylpropanoids is shown.

  In the present invention, examples of trees that are environmentally friendly algae growth inhibitors include, but are not limited to, cedar, black pine, red pine, Japanese cypress, Japanese cypress, Japanese cypress, camphor, Japanese pepper, metasequoia, and the like. Moreover, the whole, bark, heartwood, sapwood and leaves are used. The tree component of the present invention is an extract extracted from the whole or each part of the tree by steam distillation or hot water distillation, or an effluent containing the essential oil component and a solvent such as hexane, ethyl acetate, methanol, etc. Is used.

Monoterpenes (monoterpenes and their derivatives contained in tree extracts such as geraniol, hinokitiol, camphor, limonene, α and β pinene) and sesquiterpenes (longifolene, caryophyllene, elemol, cedrol) Sesquiterpenes and their derivatives contained in tree extracts such as Tyuopsen and Kazinen), diterpenes (feruginol, taxodion, 14-deoxycholone U, salvinolone, 5, 6-hydroxysgiol, etc.) Diterpenes and derivatives thereof), lignans, norlignans (hinokinin, secuirin-C, agasaresinol, sugiresinol, hydroxysgiresinol, hinokiresinol and other lignans contained in tree extracts , Norlignans and their derivatives), flavonols (flavonols and their derivatives in tree extracts such as catechin, epicatechin and taxifolin) lignin degradation products (lignin degradation produced by tree component extraction operations such as vanillin and eugenol) But not limited to these. These derivatives are not only those contained in natural tree extracts, but also esters of hydroxyl groups of the above compounds with any aliphatic organic acid or aromatic organic acid, or hydroxylamines or alkoxyamines on the carbonyl groups of the compounds. Also included are chemically synthesized derivatives such as oximes synthesized using the above.

The algae growth inhibitor of the present invention may be used as a mixture with other algae growth inhibitors, not only for prevention of red tide, but also for inhibition of algae growth in fish breeding tanks, pools, sewage treatment, etc. Is also useful. It is also useful for suppressing algae from adhering to aquaculture nets. It is also useful for controlling algal growth in closed waters such as dam reservoirs and lakes. Algae growth suppression is achieved by adding, spraying and / or applying a chemical containing the tree component of the present invention to the place where the target alga grows.

More specifically, the tree component of the present invention is mixed with a solid carrier, a liquid carrier, a gaseous carrier, and a surfactant, a sticking agent, a dispersing agent and / or a stabilizer are added as necessary, and an emulsion, It can be formulated into a wettable powder, suspension, granule, powder, dry flowable, flowable, aqueous liquid, oil, smoke, aerosol, microcapsule and the like. The above-mentioned preparation or the tree component of the present invention can be directly applied and impregnated on a fishing net or a closed wall in the aquaculture industry.

Hereinafter, the algal growth inhibitor according to the present invention will be described in more detail.

<Experiment method>
Two types of strains were used, the diatom Skeletonema costatum (NIES-16) and the raffido alga Heterosigma akashiwo (NIES-293). Both algae were purchased from the National Institute for Environmental Studies.

<Culture conditions>
The climatic device used for the culture is an NKsystem NK type artificial climatic device. The culture conditions are a temperature of 20 to 22 ° C., a light / dark cycle of 12 hours, and the light intensity is about 2000 lux during the light cycle and about 2 lux during the dark cycle. I went there. As the medium, f / 2 medium was used.
<Medium for fostering, preserving and culturing Skeletonema costatum and Heterosigma akashiwo (f / 2 medium)>
38 g of commercially available artificial seawater was added to and stirred with 1 l of distilled water, and the macronutrients shown in Table 1 were added to 1 l of the artificial seawater (f / 2). Further, trace metals shown in Table 2 were added to 1 liter of distilled water to prepare a mixed solution of trace metals (f / 2 metals), and 1 ml of f / 2 metals was added to this f / 2 and stirred to prepare an f / 2 medium. 10 ml of this f / 2 medium was dispensed into a test tube and sterilized with an autoclave. (121 degrees, 20 minutes) After sterilization, it was placed in a refrigerator and used for storage and proliferation.

<Proliferation test of Skeletonema costum and Heterosigma akashiwo>
3 to 4 ml of the f / 2 medium prepared in paragraph [0020] was placed in a test tube, and three test tubes were prepared for each concentration. Next, from the seed strain of Skeletonema costatum (NIES-16) or Heterosigma akashiwo (NIES-293), the initial concentration is about Skeletonema costatum: about 65000 to 170000 cells / ml, and about 6 to 170000 cells / ml. After inoculation, the volume was made up to 5 ml and the initial cell concentration was measured. The cells were cultured for 5 days or 7 days under the conditions shown in paragraph [0020]. Each test tube was measured every 24 hours. In Heterosigma akashiwo, when growing from a seed strain, natural growth of seawater was sterilized by autoclaving and used as a medium because growth did not occur in f / 2 medium.
In addition, a blood cell board was used for the measurement, the number of individuals in 576 cells was counted, and the number of individuals per 1 ml was converted by the following calculation formula.

(a formula)
Number of individuals per ml =
(Number of individuals in 576 trout / 576) × 1000 / 0.00025

(sample)
About 200 g to 250 g of cedar (heartwood, leaves, bark) were collected in a training forest attached to the Faculty of Agriculture, Yamagata University. About 200 g to 280 g of cypress (heartwood, leaves, bark) were collected in a training forest attached to the Faculty of Agriculture, Yamagata University. About 250 g of black pine leaves were collected at the Faculty of Agriculture, Yamagata University, and heartwood and bark were obtained from the Sakata Forestry Association.

(Collecting essential oil)
Essential oil was collected by a hot water distillation method using a hot water distillation apparatus (FIG. 2).
The collected samples were chopped with scissors to be about 1 to ² cm and 1 to ² cm ² , respectively, and 200 to 300 g of the sample was quantified. An amount of distilled water 4 to 5 times as much as the sample was placed in the round bottom flask together with the sample, and a cooler was connected to the mouth of the round flask (use a glass tube connecting part). From there, the mantle heater was set to around 150 ° C. and heated until it boiled in the round flask. Heating extraction was performed for 7 to 8 hours from the boiling point. The water vapor and volatile components which had been boiled to some extent and evaporated were passed through a cooler, and the liquid was recovered as a distillation part. After the distillation, the condenser was rinsed with ethyl acetate, and they were recovered in the same manner. The recovered material was separated into an aqueous layer and an organic layer with a separatory funnel, and only the organic layer was recovered and concentrated by an evaporator to obtain an extraction component (essential oil component). Table 3 shows the results.

(Preparation of sample for growth inhibition test)
30 mg of each essential oil collected above was weighed, 30 ml of acetone was added, and the concentration was adjusted to 1 mg / ml.

(Proliferation suppression experiment for Skeletonema costa and Heterosigma akashiwo)
The f / 2 medium prepared above was placed in a test tube with 3 to 4 ml of medium, and the concentration was adjusted to 1 ppm, 10 ppm, and 100 ppm after measuring up (5 ml) in the test tube. 5 μl, 50 μl and 500 μl of the sample were added. From there, it was left at room temperature in order to skip acetone for 2 hours. Three concentration test tubes were prepared for each. Thereafter, Skeletonema costatum (NIES-16) and Heterosigma akashiwo (NIES-293) were inoculated, and the initial cell concentration was measured. The cells were cultured under the conditions of 2-2-1 for 5 days (S. costatum) or 7 days (H. akashiwo). Each test tube was measured in the same manner as described above every 24 hours. The growth rate was calculated using the following formula.

(a formula)
Average growth rate (%)
= Number of proliferating cells in sample-added medium (number) (5th day) / sample in blank medium
Number of proliferating cells in the supplemented medium (number) (5th day) x 100

(result)
The results are shown in FIG. All essential oils showed inhibitory activity against both algae or one of them. S. The essential oil of cedar bark and black pine heartwood showed a strong growth inhibitory effect against costatum. H. In contrast to akashiwo, cedar bark, black pine heartwood / leaf / bark, and pine oil showed a strong growth inhibitory effect. In particular, the cedar bark and the essential oil of black pine heartwood showed a strong inhibitory effect against both algae.

In order to identify components that are specifically effective in suppressing the growth of the red tide algae Skeletonema costatum, 10 monoterpenes (10 carbon atoms), 5 sesquiterpenes (15 carbon atoms), diterpenes (20 carbon atoms) 11 kinds, norlignans 5 kinds, flavonols 3 kinds, lignin degradation products, lignans, quinones, tropolones, phenylpropanoids, one each, 10 kinds and 39 kinds of tree component preparations, An experiment for inhibiting the growth of the alga Skeletonema costum was performed.
The culture conditions and calculation method for the growth inhibition experiment were the same as in Example 1.

(result)
Fig. 4 shows monoterpenes, Fig. 5 shows sesquiterpenes, Fig. 6 shows diterpenes, and Fig. 7 shows growth of norlignans, flavonols, lignin degradation products, lignans, quinones, tropolones, and phenylpropanoids on Skeletonema costatum. The inhibitory effect (final average growth rate) is shown.
In general, it has activity against algae as compared to the blank, and can be a growth inhibitor by optimizing the concentration and the like.
Monoterpenes are measured for their inhibitory effect up to the third day, and can be expected for short-term effects. Sesquiterpenes showed a particularly strong inhibitory effect on longifolene. Among diterpenes and quinones, a particularly strong inhibitory effect was observed for those having specific abietatriene and quinone structures such as ferruginol and taxodion. In the inhibitory effect with flavonols, epicatechin and norlignans showed strong activity against secuirin C.

Claims (5)

An extract extracted from one or more whole trees, leaves, bark, sapwood and / or heartwood selected from the group consisting of cedar, Japanese black pine, Japanese cypress, Japanese cypress, Japanese cypress, Japanese cypress, Japanese cypress, camphor tree, raccoon, metasequoia Algae growth inhibitor contained. The algal growth according to claim 1, wherein the extract is an extract extracted with a effluent containing essential oil or an essential oil component obtained by steam distillation or hot water distillation and a solvent such as hexane, ethyl acetate, or methanol. Inhibitor. Monoterpenes wherein the extract includes i) geraniol, hinokitiol, camphor, limonene, α and β pinene,
ii) sesquiterpenes including longifolene, caryophyllene, elemol, cedrol, tyuopsen, kadinene and the like,
iii) diterpenes including ferruginol, taxodione, 14-deoxycholone U, salvinolone, 5,6-hydroxysugiol, etc.
iv) lignans and norlignans including hinokinin, secuirin-C, agasaresinol, sugiresinol, hydroxysgiresinol, hinokiresinol, etc.
v) Flavonols including catechin, epicatechin, taxifolin, etc.
vi) at least one component selected from the group consisting of lignin degradation products such as vanillin and eugenol, which are produced by tree component extraction operations, and derivatives thereof, and / or 2 The algae growth inhibitor according to Item. The algae growth inhibitor according to any one of claims 1, 2, and 3, comprising one or more of longifolene, ferruginol and derivatives thereof. The said algal growth inhibitor is effective in red tide, a sewage treatment, closed water areas, such as a dam reservoir and a lake, a fishing net, etc., The claim 1, Claim 2, Claim 3 or Claim 4 Algae growth inhibitor.