KR20050037229A - Products of complex materials for the prevention of cyanobacterial bloom - Google Patents

Abstract

The present invention relates to a composite material for inhibiting green algae generation, and more particularly, in order to suppress the growth of cyanobacteria occurring in a large amount in the summer in eutrophication water, to remove the phosphorus in the water and The present invention relates to a composite material in which a culture filtrate of Bacillus subtilis C1 [KCTC 10439BP], which selectively inhibits growth, is mixed.

Description

Products of complex materials for the prevention of cyanobacterial bloom

The present invention relates to a composite material for inhibiting green algae generation, and more particularly, in order to suppress the growth of cyanobacteria occurring in a large amount in the summer in eutrophication water, to remove the phosphorus in the water and The present invention relates to a composite material in which a culture filtrate of Bacillus subtilis C1 [KCTC 10439BP], which selectively inhibits growth, is mixed.

In recent decades, water pollution has accelerated due to the rapid increase in industrialization, urbanization, population growth, and agricultural activities. Water pollution refers to a condition in which the pollutant is discharged to the natural water and is no longer suitable for the purpose of use because it exceeds the natural purification capacity of the water, which can be classified into eutrophication and pollution by toxic substances. Recently, the most frequent and important problem among various environmental pollution will be water quality depletion of water supply by eutrophication. Eutrophication is a phenomenon in which nutrients are introduced into the water system, resulting in a decrease in algal species, and the mass growth of certain algae forms a mass of algae on the water surface. In eutrophic lakes, the amount of oxygen dissolved in the surface is usually supersaturated and the transparency becomes very low, below 1.5 m.

When the algae, ie, phytoplankton, grow in large quantities, the color of water changes, which is referred to by various terms such as hydration (algal bloom, water bloom) or green tide in freshwater. Doing. In most cases, the causative species of algae are Microcystis , Anabaena and Oscillatoria , which are taxonomic prokaryotes, which form clumps on the surface of the water, It causes taste and odor and causes various environmental problems, such as preventing sedimentation in the water purification plant.

Algal growth in most lakes depends on the amount of phosphorus and nitrogen in the nutrients, and phosphorus is known to be more important in freshwater lakes. In addition, environmental factors such as light quantity, water temperature, and the ratio of nitrogen and phosphorus have a great influence on the growth of cyanobacteria. Therefore, in order to prevent eutrophication, various methods such as removal of phosphorus, a change in algal growth conditions, etc. which play a decisive role in algal growth in freshwater lakes are required. Some chemicals, such as copper sulfate and chlorine, are used to suppress algae in eutrophic waters, but these chemicals are not specific to algae-causing algae. Algae affected by chemicals have the disadvantage of affecting the survival of ecosystem members, including toxic compounds, which make them less of a member of the food chain.

In regard to such algae removal, foreign patents related to foreign patents for the past 10 years show that there are many methods for controlling algae by chemicals. Examples of the above chemicals include peroxide (WO 01/50863), tannin (Japanese Patent No. 3161411), polymer fatty acids (US Patent No. 4398937), pigment material (US Patent No. 4781843), germanium compound (US Patent) 5518990). In addition, bio and chemical control methods using plant toxin (WO 95/12983), antibiotics (European Patent No. 0648075), viruses (Japanese Patent No. 11098979), and plant extracts (U.S. Patent No. 6149929) have been developed. Algae control methods using surfactants (Japanese Patent No. 6141726, Japanese Patent No. 9154466, US Patent No. 5407899) have also been studied.

In Korea, grain extract phytic acid (Korean Patent Registration No. 180468), electrolysis (Korean Patent Publication No. 1998-82153), porous ceramic powder (Korean Patent Registration No. 337393), electrical injury (Korean Patent Publication No. 2001-) 48041), ceramic (Korean Patent Publication No. 2000-19515), coagulation filtration (Korean Patent Registration No. 317537), glycolipid (Korean Patent Registration No. 384284), polyionic polymer (Korean Patent Registration No. 278220) Have been proposed.

However, there is a problem in such a single chemical treatment technology, such as the concern of secondary contamination, and the likelihood of only a short-term effect, there is a problem, such as an increase in the treatment cost.

It is understood that the prior art regarding composite materials using both chemical and biological materials has not been reported yet, and composite materials are expected to obtain synergistic effects by utilizing the advantages of each component and supplementing the disadvantages. Compared to the lower dose, the same effect can be obtained, and thus, the possibility of secondary pollution may be lowered, thereby enabling more environmentally friendly treatment.

Therefore, the present inventors as a composite material to more effectively suppress the algae generation, the algae growth in eutrophication system as a raw material of siliceous material which can reduce the concentration of phosphorus as determined by the concentration of phosphorus, the cyanobacteria especially microsis The present invention was completed by confirming that the filtrate of Bacillus subtilis C1, which was selectively controlled, was mixed to have an excellent effect on inhibition of cyanobacteria.

Therefore, an object of the present invention is to provide a composite material for effectively inhibiting algae generation.

The present invention is characterized by a composite material for inhibiting green algae generation treated by mixing a siliceous porous body and a culture filtrate of Bacillus subtilis C1 [KCTC 10439BP].

Referring to the present invention in more detail as follows.

The present invention relates to the use of the new strain of the prior application No. 2003-15820, in order to suppress the growth of cyanobacteria occurring in the summer in eutrophication water, to remove the phosphorus in the water of the chemicals and cyanobacteria The present invention relates to a composite material in which a culture filtrate of Bacillus subtilis C1 [KCTC 10439BP], which selectively inhibits growth, is mixed.

First, the main cause of green algae in eutrophic water systems is based on the use of siliceous materials that can effectively adsorb and precipitate phosphorus based on the fact that it is phosphorus in water. In particular, the effects of various additives are investigated to enhance the phosphorus removal ability of siliceous materials. That is, as a result of extensive investigation of the type, concentration, addition method, and the like of the additive, it was confirmed that the siliceous porous body had a particle size of 10 mm or less and a throughput of 1 to 10 g / L. At this time, when the particle size exceeds 10 mm, there is a problem that the phosphorus absorption capacity is lowered according to the relative decrease of the siliceous porous body surface area. In addition, when the amount of the siliceous porous body is less than 1 g / L, there is a problem that the phosphorus removal rate is reduced, and when it exceeds 10 g / L, there is a problem that the throughput is increased to increase the throughput. Meanwhile, in order to enhance the phosphorus removing effect of the siliceous porous body, calcium ions (Ca ²⁺ ), iron ions (Fe ³⁺ ), or both species may be additionally added, wherein calcium ions are 10 to 100 mg / L, It is preferable to add 1 to 10 mg / L of iron ions, and if the concentration is lower than this, there is almost no effect. If it exceeds this, there is a problem in that it is not suitable for using water by increasing the hardness of water.

In addition, Bacillus subtilis C1 [KCTC 10439BP], which is identified as the microorganism with the highest resolution in the genus microsis, is used as an additive for biological control. The present invention is not to use the strain directly, but to decompose the cyanobacteria using the culture filtrate in which the strain is cultured. The Bacillus subtilis C1 is cultured and the cells are removed through a centrifuge, and then the culture filtrate is recovered. The throughput of the culture filtrate is preferably maintained at 1 to 10 ml / L, and reacted at 15 to 30 ° C. for 2 to 48 hours. At this time, when the throughput is less than 1 ml / L there is a problem that the effect is insignificant, if it exceeds 10 ml / L there is a problem that the cost increases.

In order to manufacture the composite material according to the present invention, the two materials are administered in one place, but it is preferable to treat the two materials without mixing in advance, for the following reasons. The siliceous porous body contains a large amount of fine pores, and water enters the pores to adsorb phosphorus in the water. The larger the amount of pores, the more the surface area increases and the amount of phosphorus that can be adsorbed increases. Therefore, in order to further improve the phosphorus adsorption capacity of the siliceous porous body, it is completely dried and treated. However, when mixing with the Bacillus culture solution in advance, such a phosphorus adsorption effect is reduced, and thus the mixture is not treated. The order of treatment is a siliceous porous body, Bacillus culture medium, the time interval of the two treatments can be changed depending on the conditions of the pond or lake to be treated and the effect to be obtained. For example, in the case of high phosphorus concentration and severe green algae, the Bacillus culture medium is treated immediately after the siliceous porous body treatment, and when the green algae are not near, only the siliceous porous body is treated and the Bacillus culture liquid is imminent when the green algae occurs. Is the most efficient.

These composites have the following characteristics.

First, the chemical material is an effective way to prevent algal blooms in advance by focusing on the removal of phosphorus, which is a major element of algal growth. Second, the biological material has an excellent resolution of the genus microcistis in the microalgae, so that it can be selectively decomposed and removed, and thus it is possible to maximize the conservation of the aquatic ecosystem because it does not affect other harmless algae.

Therefore, the composite material can achieve ecological restoration of the environment by regulating cyanobacteria on the basis of physiological and ecological characteristics of green algae in eutrophic waters.

Hereinafter, the present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1 Selection of Silicate Porous Material

Siliceous porous material is a porous calcium silicate compound prepared by synthesizing raw materials such as gypsum, quicklime and steelmaking slug. The main chemical components are SiO ₂ and CaO, and some oxides of Al, Fe, Mg and K Included.

Chemical Composition and Physical Properties of Silicate Porous Body Chemical composition (%) SiO ₂ 45 to 55 CaO 25 to 35 Al ₂ O ₃ 3 to 4 Fe ₂ O ₃ 1 to 2 MgO 1 to 2 K ₂ O 0.5 to 1.0 Physical properties pH 8 to 9 importance 0.35-0.45 Specific surface area 50 m ² / g Water absorption 1.5 times of self-water

Distilled water was used to investigate the effect of phosphorus removal on various conditions of siliceous porous body. The stationary sample was prepared by dissolving KH ₂ PO ₄ in distilled water, and the change in phosphorus concentration according to the particle size and throughput of the siliceous porous body was investigated over time.

1) Phosphorus removal effect by particle size of siliceous porous body

Silicate porous bodies having a particle size of 1, 2 and 4 mm or less were added at 10 g / L, and the phosphorus concentration of all samples including the control (Control) was adjusted to 0.26 mg / L and the pH to 7. Phosphorus concentration continued to decrease with time [FIG. 1]. After 7 days, the lowest concentration was 0.05 mg / L at the particle size of 1 mm or less and the phosphorus concentration was 0.06 mg / L at the particle size of 2 mm or less and 4 mm or less. According to two-way ANOVA, the phosphorus removal effect of the siliceous porous body was not different between 2 mm and 4 mm in particle size ( F _(1,8) = 0.05, P = 0.829), but 1 mm in particle size. Less than 2 mm ( F _(1,8) = 12.4, P = 0.008), 4 mm or less ( F _(1,8) = 11.24, P = 0.010) was significantly different. Since the larger the specific surface area, the larger the contact surface reacting with phosphorus in water, the siliceous porous body having a specific surface area of 1 mm or less having the same specific weight at the same weight was evaluated as the best in the phosphorus removal effect.

2) Phosphorus removal effect according to throughput of siliceous porous body

Silicate porous bodies having a particle size of 1 mm or less were added to the Erlenmeyer flasks at 1, 2.5, 5 and 10 g / L, respectively. Phosphorus concentration was adjusted to 0.22 mg / L and pH to 7, respectively, to investigate the change over time in phosphorus concentration at room temperature for 7 days.

As a result, the larger the throughput of the siliceous porous body, the more drastically the decrease in phosphorus concentration was [Fig. 2]. Phosphorus concentration in the treatment group to which 10 g / L of siliceous porous body was added decreased to 0.04 mg / L after 7 days from the initial 0.22 mg / L. Phosphorus removal effect of the siliceous porous body was proportional to the increase of the addition amount.

Example 2: Enhancement of Phosphorus Removal by Additives

The effect of phosphorus removal by Ca and Fe compound addition with siliceous porous body was investigated over time. One control (A) and four treatments (B: siliceous porous body, C: siliceous porous body + Ca, D: siliceous porous body + Fe, E: siliceous porous body + Ca + Fe) were each 150 ml in a 250-ml Erlenmeyer flask. Distilled water and the siliceous porous body of 1 mm or less of particle size included 1 g / L. Phosphorus concentrations in the control and treatment were adjusted to 0.10 and 1.0 mg / L using K ₂ HPO ₄ , respectively, and the concentrations of Ca and Fe in the treatment were CaCl ₂ ㆍ 2H ₂ O and FeCl ₃ ㆍ 6H ₂ O. 30 mg / L and 1 mg / L, respectively, were adjusted. Initially, phosphorus concentrations were measured for a total of 8 days at 2, 4, 8, 12 hour intervals, and then at 2 day intervals.

As a result of investigating the change of phosphorus concentration after adding the siliceous porous body, Ca and Fe compounds single or in combination, the phosphorus of treatment E (silicate porous body + Ca + Fe) within 2 hours after adding 0.10, 1.0 mg / L phosphorus The concentrations decreased sharply to 0.01 and 0.54 mg / L, respectively [FIG. 3]. In addition, three different treatments B, C, D also showed a decrease in phosphorus concentration compared to the control. Afterwards, the phosphorus concentration in the treatment group tended to increase again until 12 hours, but between 2 and 8 days after treatment, the phosphorus concentration was similar to or gradually decreased after the initial 12 hours. At the end of the experiment, the phosphorus concentration of treatment group E was examined at 0.03 and 0.47 mg / L, respectively, to record the lowest phosphorus concentration. Therefore, the phosphorus removal effect was found to be enhanced by adding Ca and Fe compounds together than when only siliceous porous bodies were treated.

Example 3 Growth Inhibition and Removal of Microcistis by Bacillus Culture Filtrate

Microcystis aeruginosa was transferred to Allen liquid medium [J. Phycol., Allen, MM 1968. 4: 1-4] were incubated at about 28 ℃, 100 μmol / m ² / s, 100 rpm. After the exponential growth period, the Bacillus subtilis C1 culture filtrate was added at a concentration of 10 ml / L in a very high concentration of microsistis to observe the difference in growth. The experiment was started at the point of chlorophyll- a concentration of about 1,000 ㎍ / L and observed the change in the concentration of chlorophyll- a for two days after treatment.

As shown in FIG. 4, at least 60% of the microcistis was removed after one day of treatment, and at least 85% after 2 days. After treating the culture filtrate of Bacillus subtilis C1, the culture medium of microcistis, which had a dark green color, was discolored and turned white in about 2 to 3 hours, and the death of cyanobacteria was easily seen with the naked eye.

Example 4 Preparation of Composite

50 mg / L of CaCl ₂ ㆍ 2H ₂ O and 5 mg / L of FeCl ₃ · 6H ₂ O are sequentially mixed with 5 g / L siliceous porous body having a particle size of 10 mm or less and incubated at 25 ° C. for 3 days immediately before use in the field. 5 ml / L of the Bacillus subtilis C1 filtrate was prepared to prepare a composite material.

Example 5 Confirmation of Green Algae Inhibition Effect of Composite Material

Into a 250-mL Erlenmeyer flask, add 100 ml of Daecheong water (filtered water, filtered or sterilized) or Allen medium, inoculate Microcystis aeruginosa, and incubate for 8 days to incubate the type and density of algae over time. Investigate. There were five experimental zones: control (Daechung) without adding microcistises to Daecheongho raw water, treatment (Daechung + M. aeruginosa ) inoculating microcistisate to Daecheongho raw water, and microcistis to filtrate of Daecheongho raw water. One treatment group (Filtrated Daechung + M. aeruginosa ), one treatment group autoclaved Daechung + M. aeruginosa and autoclaved Daechung + M. aeruginosa , and the other group treatment group (Allen + M. aeruginosa) )to be. Cultivation of these five experimental zones was carried out by dividing into two environments: shaking incubator conditions with a brightness of 150 μmol / m ² / s and a temperature of 30 ° C. adjusted to outdoor natural light conditions and similar environments. The reason why the experiment was carried out under these two conditions is that the effect is usually determined by using only the experimental results performed in the laboratory. However, in actual natural environmental conditions, the results are often different. The purpose of this study was to compare the results of the two culture conditions.

Outdoor natural light cultivation was carried out on the roof of the Institute in August 2003. During this period, the average temperature was about 23 ° C, and the temperature of the culture medium in the flask was about 30 ° C at midday, similar to that of the shaker.

As shown in Figure 5, only the addition of "Daecheongho water" and "Daecheongho water + M. aeruginosa " of the composite material treatment prepared in Example 4 only increased in vivo fluorescence, other treatments There was little growth of algae in the two treatments, but few other algal species were found, with Chlamydomonas dominant. In other words, since the growth occurred only in the treatment group in which Chlamydomonas existed at the early stage of culture, the green algae Clamydomonas was not affected by the composite material, and the microalgae microsis was completely inhibited by the composite material. have. On the other hand, the addition of only Daecheongho water and the addition of Daecheongho water + M. aeruginosa in the composites showed that a large variety of algae, including microsis, proliferated.

From the above results, the composites selectively inhibit growth of cyanobacteria, especially microcistis, which cause harmful environmental problems, but do not affect algae, such as Chlamydomonas, which do not cause problems. It is expected to enable environmentally friendly algae control without destroying the ecosystem.

As described above, the composite material prepared in accordance with the present invention to suppress the development of green algae at the same time to suppress the algae generation by the removal of phosphorus in the water by the chemical material, at the same time to suppress the selective growth of algae species causing green algae caused by biological material It is environmentally friendly as it can be achieved and is expected to be very useful for ecosystem restoration.

1 is a graph showing the effect of removing phosphorous (phosphorus) by the particle size of the siliceous porous body.

Figure 2 is a graph showing the phosphorus removal effect according to the addition amount of the siliceous porous body.

3 is a graph showing the phosphorus removal effect according to the addition of cations to the siliceous porous body [a: initial phosphorus concentration 0.1 mg-P / L, b: initial phosphorus concentration 1 mg-P / L].

Figure 4 is a graph showing the growth inhibition of Microcystis sp. According to the addition of Bacillus subtilis C1 (KCTC 10439BP) culture filtrate.

Figure 5 is a graph showing the effect of inhibiting the growth of microorganism genus of the composite material in the incubator and natural light conditions.

Claims (4)

A composite material for inhibiting green algae generation, characterized by treating a mixture of a siliceous porous body and a culture filtrate of Bacillus subtilis C1 [KCTC 10439BP]. The composite material according to claim 1, wherein the siliceous porous body has a particle size of 10 mm or less and a content of 1 to 10 g / L. The composite material for inhibiting green algae generation according to claim 1, wherein calcium ions (Ca ²⁺ ) or iron ions (Fe ³⁺ ) are further added to the composite material to enhance the phosphorus removal effect of the siliceous porous body. . According to claim 1, wherein the Bacillus subtilis C1 culture filtrate content of 1 to 10 ㎖ / L composite material for inhibiting green algae generation.