KR102127466B1 - Algicidal Composition for Controlling Freshwater Algae - Google Patents
Algicidal Composition for Controlling Freshwater Algae Download PDFInfo
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- KR102127466B1 KR102127466B1 KR1020190016905A KR20190016905A KR102127466B1 KR 102127466 B1 KR102127466 B1 KR 102127466B1 KR 1020190016905 A KR1020190016905 A KR 1020190016905A KR 20190016905 A KR20190016905 A KR 20190016905A KR 102127466 B1 KR102127466 B1 KR 102127466B1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/36—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a singly bound oxygen or sulfur atom attached to the same carbon skeleton, this oxygen or sulfur atom not being a member of a carboxylic group or of a thio analogue, or of a derivative thereof, e.g. hydroxy-carboxylic acids
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
Abstract
Description
본 발명은 고가의 장비를 사용하지 않고, 미생물의 병원성이나 살조물질의 독성에 의한 환경오염이나 생태계 교란의 문제 없이 담수 녹조를 방제할 수 있는 살조 조성물에 관한 것이다. The present invention relates to an algae composition that can control freshwater green algae without the problem of environmental pollution or ecological disturbance due to pathogenicity of microorganisms or toxicity of algae substances without using expensive equipment.
지구 온난화로 인한 기온 상승과 강수량의 저하는 저수지나 호수와 같은 담수에 유해조류 대발생(Harmful aglae bloom)을 야기하고 있다. 유해조류 대발생은 수생생물의 폐사를 일으켜 생태계를 교반시키며, 음용수 및 농업용수 자원의 확보 및 관리를 어렵게 한다. 그뿐 아니라 유해조류 대발생의 주요 원인 생물인 Anabaena flos-aquae, Microcystis aeruginosa 등의 남조류가 생성하는 간독소(microcystin)는 동물 및 사람에게 치명적인 해를 주는 것으로 알려져 있다.Due to global warming, the rise in temperature and the decrease in precipitation are causing harmful aglae blooms in fresh water such as reservoirs and lakes. The generation of harmful algae causes the death of aquatic organisms, stirs the ecosystem, and makes it difficult to secure and manage drinking water and agricultural water resources. In addition , microcystin produced by blue-green algae such as Anabaena flos-aquae and Microcystis aeruginosa, which are major causes of harmful algae outbreaks , is known to cause fatal harm to animals and humans.
유해 조류 방제방법은 화학적 방제, 물리적 방제 및 생물학적 방제로 나눌 수 있다. 화학적 방제는 황산동, 이산화염소, 시마진(Simazine) 등의 약품을 살포하는 기술로 그 효과는 비교적 우수하나 경제적 비용이 크고, 비선택적이며, 특히 일부 성분의 경우 중금속이 축적되거나 유기물질과 상호작용하여 유독물질을 생성할 가능성이 있다. 물리적 방제(공개특허 제10-2011-0026204)는 정수처리에 의한 방제로 응집, 침전, 여과, 활성탄 흡착, 염소 소독 및 오존산화 등을 적용하는 기술로, 펌프 등의 순환 시스템의 설치 운용비용 및 화학 약품비 등 고가의 비용 문제로 인하여 실제로 적용하기에는 많은 어려움이 있다. 생물학적 방제는 수생식물, 미생물, 동물성 플랑크톤, 어패류와 같은 생물 조절인자들을 수계에 적용시켜 제어하는 기술로, 이 중 살조세균을 이용한 제어방법이 많이 연구되었다(등록특허 제10-1311837호, 등록특허 제10-1661543호). 그러나, 대부분의 살조세균은 병원성 세균으로서 현장적용 시 수계환경의 다른 생물에 대한 병원성 문제와 2차 교란이 발생할 수 있고, 살조세균의 적용 시기 및 농도에 따라 그 효율이 매우 다르게 나타나며, 또한 과량의 미생물 주입 시 배양에 사용한 질소·인의 함량이 높은 배지가 오히려 조류의 성장을 더욱 촉진시키는 결과를 가져올 수 있다. 이러한 문제를 해결하고 현장에서 친환경적이며 효율적으로 조류를 제어하기 위해서는 살조세균이 조류의 표면에 접촉하여 세포를 요해시키는 직접 공격형 기작이 아닌, 그들이 체외로 분비하는 물질을 적용하는 간접적 제어기작의 활용이 필요하다(등록특허 제10-0495271호). 등록특허 제10-1311837호는 특정 락토바실러스 속 또는 락토코커스 속 세균들이 녹조를 포함하는 유해조류에 대하여 우수한 살조 효과가 있음을 보고하였다. 유산균은 종래 살조 효과를 갖는 살조세균들이 병원성으로 인해 다른 생물의 생존을 위협하고, 생태계를 교란시키는 문제를 갖는 것과는 달리 안전하게 사용할 수 있다는 장점이 있다. 그러나, 병원성 살조세균들에 비해 살조활성이 낮기 때문에 살조 효과가 미흡하다는 문제가 있었다. Methods for controlling harmful algae can be divided into chemical control, physical control, and biological control. Chemical control is a technology that sprays chemicals such as copper sulfate, chlorine dioxide, and simazine, but its effect is relatively good, but it is economically expensive and non-selective. In particular, heavy metals accumulate or interact with organic substances Thereby, it is possible to produce toxic substances. Physical control (published patent No. 10-2011-0026204) is a technology that applies aggregation, sedimentation, filtration, activated carbon adsorption, chlorine disinfection and ozone oxidation as a control by water treatment, and installs and operates the circulation system of pumps, etc. Due to the high cost of chemicals and other cost problems, there are many difficulties in practical application. Biological control is a technology that controls by applying biological regulators such as aquatic plants, microorganisms, zooplankton, and fishery products to aquatic systems, and among them, a lot of methods of controlling using alginate bacteria have been studied (Registration Patent No. 10-1311837, registered patent) No. 10-1661543). However, most algae bacteria are pathogenic bacteria, and when applied in the field, pathogenic problems and secondary disturbances to other organisms in the aquatic environment may occur, and their efficiency is very different depending on the application time and concentration of the algae bacteria. When injecting microorganisms, a medium having a high content of nitrogen and phosphorus used for cultivation may result in further promoting algae growth. In order to solve these problems and to control the algae in an environment-friendly and efficient manner in the field, the use of an indirect control mechanism that applies substances that they secrete outside the body is not a direct attacking mechanism in which algae bacteria contact the surface of the algae and damage the cells. It is necessary (Registration Patent No. 10-0495271). Patent No. 10-1311837 reported that bacteria of the genus Lactobacillus or Lactococcus have an excellent algicidal effect against harmful algae including green algae. Lactic acid bacteria have the advantage that they can be safely used, unlike conventional algebra bacteria having an agicide effect threaten the survival of other organisms due to the pathogenicity and have a problem of disturbing the ecosystem. However, there is a problem that the algebraic effect is insufficient because the algebraic activity is lower than that of the pathogenic agar bacteria.
본 발명은 병원성이 없어 안전하지만 살조활성이 다소 낮은 유산균에 시너지 효과를 나타낼 수 있는 성분을 혼합하여 우수한 살조활성을 나타내도록 한 담수 녹조 방제용 살조 조성물을 제공하는 것을 목적으로 한다. An object of the present invention is to provide an algae composition for controlling freshwater green algae by mixing components capable of exhibiting a synergistic effect on lactic acid bacteria which are safe because they have no pathogenicity, but which have slightly lower algae activity.
전술한 목적을 달성하기 위한 본 발명은 유산균과 유기산 및 유지의 혼합물을 유효성분으로 함유하는 담수 녹조 방제용 살조 조성물에 관한 것이다. The present invention for achieving the above object relates to a freshwater algae control algae composition containing a mixture of lactic acid bacteria, organic acids and fats and oils as an active ingredient.
유산균과 유기산 혹은 유산균과 유지의 혼합물은 유산균 단독에 비해 특이할만한 시너지 효과를 나타내지 못하였으나, 유산균과 유기산 및 유지의 혼합물은 시너지 효과로 인해 살조활성이 크게 증가하여 대표적인 살조세균인 Chitinimonas koreensis KACC 11467보다도 우수한 살조 효과를 나타내었다. The mixture of lactic acid bacteria and organic acids or lactic acid bacteria and fats and oils did not show a specific synergistic effect compared to lactic acid bacteria alone, but the mixture of lactic acid bacteria and organic acids and fats significantly increased the algebraic activity due to the synergistic effect, than the typical algal bacteria Chitinimonas koreensis KACC 11467 It showed an excellent algal effect.
상기 유산균으로는 락토바실러스 가사리, 락토바실러스 카세이, 락토바실러스 플란타럼, 루코노스톡멘센테로이드스, 락토바실러스 부크너리, 락토바실러스 파라카세이 및 락토바실러스 하비넨시스로 이루어진 군으로부터 선택된 하나 이상인 것이 바람직하였으며, 특히 락토바실러스 부크너리인 것이 더욱 바람직하였다.The lactic acid bacteria are preferably at least one selected from the group consisting of Lactobacillus gasari, Lactobacillus casei, Lactobacillus plantarum, Lucono stock mencentroids, Lactobacillus Buchnery, Lactobacillus paracasei, and Lactobacillus habinensis. In particular, it was more preferable to be Lactobacillus Buchnery.
상기 유산균의 접종 농도는 통상 102~1010 cells/ml인 것이 바람직하며, 유해조류의 발생 정도 및 목적(예방 또는 제거), 유산균의 종류에 따라 적절한 접종농도로 투여할 수 있다.The inoculation concentration of the lactic acid bacteria is preferably 10 2 ~ 10 10 cells/ml, and can be administered at an appropriate inoculation concentration according to the degree and purpose (prevention or removal) of harmful algae, and the type of lactic acid bacteria.
본 발명에서 유기산은 산성을 나타내는 유기 화합물을 의미하는 것으로, 유산균과의 공동처리에 의한 사전 스크리닝에 의하면, 숙신산, 시트르산 및 탄닌산인 것이 바람직하였다. 유기산의 처리농도는 유기산의 종류에 따라 미차가 있기는 하였으나, 0.5~3중량%인 것이 바람직하였다. 유기산의 농도가 너무 낮으면 유기산의 첨가로 인한 효과가 충분하지 않았으며, 유기산의 농도를 더 증가시킨다고 하더라도 살조 효능의 증가는 크지 않아 경제성이 낮았다.In the present invention, the organic acid means an organic compound showing acidity, and according to pre-screening by co-treatment with lactic acid bacteria, it was preferable that it was succinic acid, citric acid and tannic acid. The treatment concentration of the organic acid was slightly different depending on the type of the organic acid, but was preferably 0.5 to 3% by weight. When the concentration of the organic acid was too low, the effect due to the addition of the organic acid was not sufficient, and even if the concentration of the organic acid was further increased, the increase in the effectiveness of the algebra was not large, resulting in low economic efficiency.
본 발명에서 유지란 동물 또는 식물에서 채취한 기름을 총칭하는 것으로, 동물성 유지는 포화지방산을 다량 함유하여 고체상태인 경우가 많기 때문에 식물성 유지를 사용하는 것이 더 바람직하였다. 식물성 유지로는 대두유, 해바라기씨유, 포도씨유, 카놀라유, 올리브유 등을 예로 들 수 있으나, 이에 한정되는 것은 아니다. 유지의 처리농도는 식물성 유지의 경우에는 유지의 종류에 거의 영향을 받지 않았으며, 0.5~3중량%인 것이 바람직하였다. 유지의 농도가 너무 낮으면 유지의 첨가로 인한 효과가 충분하지 않았으며, 유지의 농도를 더 증가시킨다고 하더라도 살조 효능의 증가는 크지 않아 경제성이 낮았다.In the present invention, the oil and fat is a generic term for oil collected from animals or plants, and it is more preferable to use vegetable oil because animal fats and oils often contain a large amount of saturated fatty acids and are in a solid state. Vegetable oils include, but are not limited to, soybean oil, sunflower seed oil, grape seed oil, canola oil, olive oil, and the like. In the case of vegetable oil and fat, the treatment concentration of the oil and fat was hardly affected by the type of oil and fat, and it was preferable that it was 0.5 to 3% by weight. If the concentration of fat and oil was too low, the effect of adding fat and oil was not sufficient, and even if the fat and oil concentration was further increased, the increase in the effectiveness of algebra was not large, and the economic efficiency was low.
본 발명의 조성물은 유해조류의 발생이 빈번하거나 발생징후가 관측된 지역에 예방적으로 처리하거나, 혹은 이미 유해조류가 발생된 지역에서 유해조류의 제거를 위하여 국지적으로 처리하는 것에 의해 담조 녹조를 방제할 수 있다. 상기 조성물은 조성물 자체로 살포하거나 혹은 별도의 담체에 고정화하여 처리할 수 있다. 본 발명의 조성물에 대해 방제의 목적이나, 유해조류의 발생정도, 유해조류의 종류, 각 조성물의 성분 등을 고려하여 최선의 효과를 낼 수 있는 농도를 설정하는 것은 당업자에게 용이할 것이다. The composition of the present invention prevents algal algae by treating prophylactically in areas where frequent occurrence of harmful algae or signs are observed, or by locally treating for the removal of harmful algae in areas where harmful algae have already occurred. can do. The composition may be treated by spraying with the composition itself or immobilized on a separate carrier. It will be easy for a person skilled in the art to set the concentration that can produce the best effect in consideration of the purpose of control, the degree of occurrence of harmful birds, the type of harmful birds, the components of each composition, etc. for the composition of the present invention.
이상과 같이 본 발명의 조성물에 의하면 조성물에 함유된 세균이 병원성을 갖지 않고, 살조물질이 독성을 나타내지 않으며 각 성분들의 혼합에 의해 시너지 효과를 나타내어 살조활성이 우수하여 담수 녹조의 방제에 유용하게 사용될 수 있다. As described above, according to the composition of the present invention, the bacteria contained in the composition do not have pathogenicity, the algae material does not exhibit toxicity, and it exhibits a synergistic effect by mixing each component, and thus has excellent algicidal activity, and thus can be usefully used for controlling freshwater green algae. Can.
도 1은 디스크법에 의한 유산균의 살조활성 분석 실험 결과를 보여주는 사진.
도 2는 액체배양법에 의한 유산균의 살조활성 분석 실험 결과를 보여주는 사진.Figure 1 is a photograph showing the results of the analysis of the analytic activity of lactic acid bacteria by the disk method.
Figure 2 is a photograph showing the experimental results of the analysis of algae activity of lactic acid bacteria by liquid culture method.
이하 첨부된 실시예를 들어 본 발명을 보다 상세히 설명한다. 그러나 이러한 실시예는 본 발명의 기술적 사상의 내용과 범위를 쉽게 설명하기 위한 예시일 뿐, 이에 의해 본 발명의 기술적 범위가 한정되거나 변경되는 것은 아니다. 이러한 예시에 기초하여 본 발명의 기술적 사상의 범위 안에서 다양한 변형과 변경이 가능함은 당업자에게는 당연할 것이다. Hereinafter, the present invention will be described in more detail with reference to the attached examples. However, these examples are merely examples for easily explaining the contents and scope of the technical spirit of the present invention, and the technical scope of the present invention is not limited or changed thereby. 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.
[실시예][Example]
실시예 1 : 남조류의 배양Example 1: Culture of blue-green algae
실험 조류인 남조세균 Anabaena flos-aquae는 한국생명공학연구원 생물자원센터(Korean Collection for Type Culture, KCTC AG10011)로부터 분양받았으며, Anabaena sp. MH-02는 국내 대청호에서 직접 분리하였다. 남조류 배양은 BG11 배지(Allen and Stainer 1968)를 이용하였다. 배양조건은 50 μmol photons/m2·s, 25℃, 12 : 12 (light : dark) cycle의 조건으로 1~2주간 정치배양한 후, 그 일부를 접종균으로 새로운 BG11 배지에 접종하여 사용하였다. Anabaena flos-aquae , a blue - green algae, was purchased from the Korean Institute of Biotechnology and Biotechnology Resource Center (Korean Collection for Type Culture, KCTC AG10011). Anabaena sp. MH-02 was directly separated from Daecheong Lake in Korea. Blue-green algae culture was performed using BG11 medium (Allen and Stainer 1968). The culture conditions were 50 μmol photons/m 2 ·s, 25° C., 12: 12 (light: dark) cycle, and then incubated for 1 to 2 weeks, a part of the culture was inoculated into a new BG11 medium as an inoculum. .
실시예 2 : 유산균의 배양Example 2: Culture of lactic acid bacteria
하기 표 1의 유산균은 KCTC로부터 분양 받거나, 또는 자체 분리한 균을 이용하였다. 유산균은 유산균 배지인 MRS broth(deMan et al., 1960)에서 36℃, 250 rpm으로 교반배양하였다. 동결보존을 위하여 배양, 농축한 세포는 25% glycerol이 포함된 MRS 배지에 옮겨 -80℃에서 냉동 보관하였다. The lactic acid bacteria in Table 1 below were pre-sold from KCTC, or used as their own isolates. Lactic acid bacteria were stirred and cultured at 36°C and 250 rpm in MRS broth (deMan et al., 1960), which is a lactic acid bacteria medium. Cells cultured and concentrated for cryopreservation were transferred to MRS medium containing 25% glycerol and stored frozen at -80°C.
실시예 3 : 유산균의 살조활성 평가Example 3: Evaluation of algicide activity of lactic acid bacteria
유산균은 동결보존한 stock으로부터 MRS 1.5% agar 평판 배지에 선조접종 (streaking)하여 얻은 단일 콜로니(single colony)를 5 mL Lactobacilli MRS broth를 이용하여 36℃, 300 rpm 24시간 액체 배양하였다. 배양액을 12,000 rpm, 10분 동안 원심분리하여 세포침전물을 회수한 후, 남조류의 성장 배지인 멸균된 BG11 배지로 2번 세척하고 세포수를 1.0 × 108 cells/mL(OD600nm = 1.2)로 조절하였다. Lactic acid bacteria were cultured at 36° C. and 300 rpm for 24 hours using 5 mL Lactobacilli MRS broth for single colonies obtained by streaking MRS 1.5% agar plate medium from freeze-preserved stock. After centrifuging the culture solution for 12,000 rpm for 10 minutes to recover the cell precipitate, the cells were washed twice with sterile BG11 medium, a growth medium of blue-green algae, and the cell number was adjusted to 1.0 × 10 8 cells/mL (OD 600nm = 1.2). Did.
실험 조류인 Anabaena flos-aquae는 BG11 배지에 접종한 후 7일간 25℃, 50 μmol photons/m2·s, 12 : 12 (light : dark) cycle의 조건으로 정치 또는 교반 배양하면서 혈구계산기(haemocytometer)로 계수하거나, 흡광도 1.0 × 104 cells/mL (OD600nm = 0.2) 수준까지 배양하였다. Anabaena flos-aquae , an experimental algae, was inoculated in BG11 medium and cultured under static or agitated conditions at 25°C, 50 μmol photons/m 2 ·s, 12: 12 (light: dark) cycle for 7 days, followed by a hemocytometer. Or cultured to absorbance of 1.0 × 10 4 cells/mL (OD 600nm = 0.2).
1) 디스크법에 의한 살조효능 평가1) Evaluation of algae efficacy by disc method
배양한 실험조류 0.2 mL를 BG11 agar 배지에 도말하고 1~2일 배양한 후 배지 위에 멸균 종이 디스크를 올려놓았다. 1.0 × 108 cells/mL로 세포수를 조절한 유산균 배양액 30 μl를 종이 디스크 위에 서서히 흡착시킨 후, 남조류 배양조건에서 7일간 배양하면서 형성된 투명환의 크기로 남조류의 생육저해율을 측정하였다. 비교를 위하여 종래기술에서 살조효능이 있는 것으로 알려진 Pichia kudriavzevii RPG-Y0002(Y2), Pseudomonas fluorescens KACC 10327, Pseudomonas fluorescens KACC 12332, Chitinimonas koreensis KACC 11467에 대해서도 동일한 방법에 의해 생육저해율을 측정하였다.0.2 mL of the cultured experimental algae was spread on BG11 agar medium, incubated for 1 to 2 days, and a sterile paper disk was placed on the medium. 30 μl of the lactic acid bacteria culture solution with cell number adjusted to 1.0×10 8 cells/mL was slowly adsorbed onto a paper disc, and the growth inhibition rate of blue-green algae was measured with the size of a transparent ring formed while culturing for 7 days in blue-green algae culture conditions. For comparison, the growth inhibition rate was measured by the same method for Pichia kudriavzevii RPG-Y0002 (Y2), Pseudomonas fluorescens KACC 10327, Pseudomonas fluorescens KACC 12332, Chitinimonas koreensis KACC 11467, which are known to have an algicide effect in the prior art.
도 1은 디스크법에 의한 유산균의 살조활성 분석 실험 결과를 보여주는 예시적인 사진으로, 유산균의 종류에 따라 남조류의 생육저해능에 차이를 보임을 알 수 있다. 이에 저해거리 3~4 mm 이상의 저해효과를 보인 유산균 12종을 2차 선발하여 표 2에 기재하였다. 표 2에서 확인할 수 있듯이 Lactobacillus buchneri RPG-L0001(L1)의 남조류 생육저해능이 가장 높아 대표적인 살조균주로 알려진 Chitinimonas koreensis KACC 11467와 유사한 수준의 활성을 보였다. 표 2에서 살조활성의 기호는 다음을 의미한다; +: 1-2mm; ++: 2-3mm; +++: 3-4mm; ++++: 4-5mm.Figure 1 is an exemplary photo showing the results of the analysis of the anaerobic activity of lactic acid bacteria by the disc method, it can be seen that the difference in growth inhibition ability of blue-green algae according to the type of lactic acid bacteria. To this end, 12 types of lactic acid bacteria that exhibited an inhibitory effect of 3-4 mm or more were secondarily selected and are listed in Table 2. As can be seen in Table 2, Lactobacillus buchneri RPG-L0001 (L1) showed the highest level of cyanobacteria growth inhibitory activity, showing a similar activity to that of C hitinimonas koreensis KACC 11467, which is known as a representative fungicide. In Table 2, the symbol of algicide activity means: +: 1-2 mm; ++: 2-3 mm; +++: 3-4mm; ++++: 4-5mm.
2) 액체배양법에 의한 살조효능 평가2) Evaluation of algae efficacy by liquid culture method
디스크법에 의해 2차 선발한 유산균의 살조효능을 액체배양법에 의해 평가하였다. 배양한 선발 유산균(1.0 × 108 cells mL-1) 0.1 mL를 9.9 mL BG11 배지에서 Anabaena flos-aquae (1.0 × 104 cells mL-1)가 전배양된 시험관에 접종 후 7 일째에 남조류의 세포수를 광학현미경 하에서 혈구계산기 또는 분광광도계를 이용하여 OD값(660nm)을 측정하였다. 유산균의 살조활성(%)은 아래 식을 사용하여 남조류 세포의 감소율을 계산하였다.The killing efficacy of the lactic acid bacteria selected second by the disk method was evaluated by the liquid culture method. 0.1 mL of cultured selected lactic acid bacteria (1.0 × 10 8 cells mL -1 ) into a test tube pre-cultured with Anabaena flos-aquae (1.0 × 10 4 cells mL -1 ) in 9.9 mL BG11 medium 7 days after inoculation The OD value (660 nm) was measured using a hemocytometer or a spectrophotometer under an optical microscope. The algebraic activity (%) of lactic acid bacteria was calculated by using the following formula to reduce the rate of cyanobacteria cells.
살조활성 (%) = (1 - Tt/Ct) × 100 Algal activity (%) = (1-T t /C t ) × 100
T: 시간 t에서 처리구의 조류세포 밀도 T: algal cell density of the treatment at time t
C: 시간 t에서 대조구의 조류세포 밀도 C: Algal cell density of the control at time t
도 2는 액체배양법의 실험 사진이며, 표 3에 접종 7일 후의 살조효능을 기재하였다. 표 3은 디스크법과 마찬가지로 액체배양법에서도 Lactobacillus buchneri RPG-L0001(L1)의 남조류 생육저해능이 가장 높아 대조군에 비해 약 53% 정도의 세포밀도 저하 효과를 나타냄을 보여준다. 대표적인 살조균주인 Chitinimonas koreensis KACC 11467은 약 60%의 억제 효능을 나타내었다.Figure 2 is an experimental picture of the liquid culture method, Table 3 describes the algebra efficacy after 7 days of inoculation. Table 3 shows that Lactobacillus buchneri RPG-L0001(L1) has the highest growth inhibitory ability of blue-green algae in the liquid culture method, as compared to the control group. It shows that it shows the effect of reducing cell density by about 53%. A representative fungal strain, C hitinimonas koreensis KACC 11467, showed an inhibitory effect of about 60%.
배양 기간별 유산균의 살조활성을 분석하기 위하여, 상기와 같은 조건에서 유산균 접종 후 1, 2, 3, 7일의 살조활성을 분석하고 그 결과를 표 4에 기재하였다. 대조구에 비해 유산균 접종구는 배양 5일 후까지 대략 20% 내외의 살조활성을 나타내었으나, 배양 7일 후에는 대부분의 접종구들은 40% 이상의 살조활성을 나타내었다. Lactobacillus buchneri RPG-L0001(L1)은 5일 후 34%, 7일 후 57%의 살조활성을 나타내어 살조활성이 가장 우수하였다.In order to analyze the algebraic activity of lactic acid bacteria by culture period, the algebraic activity of 1, 2, 3, 7 days after lactic acid bacteria inoculation under the above conditions was analyzed and the results are shown in Table 4. Compared to the control, the lactic acid bacteria inoculation showed approximately 20% of the algae activity until 5 days after culture, but after 7 days of culture, most of the inoculation showed more than 40% of the algae activity. Lactobacillus buchneri RPG-L0001 (L1) showed the highest algae activity, showing 34% after 5 days and 57% after 7 days.
실시예 4 : 유산균과 살조물질 혼합물의 살조활성 평가Example 4: Evaluation of algicide activity of a mixture of lactic acid bacteria and algicide
최종 선발된 유산균 L. buchneri L1과 유기산인 Succinic acid 및/또는 식물성 유지인 Soybean oil의 혼합물의 살조활성을 분석하여 표 5와 표 6에 그 결과를 기재하였다. Analysis of the apricot activity of the mixture of the finally selected lactic acid bacteria L. buchneri L1 and organic acid Succinic acid and/or vegetable oil Soybean oil is described in Tables 5 and 6.
하기 표 5로부터 유산균에 Succinic acid 또는 Soybean oil 첨가 시 살조활성이 각각 62%와 64%로 유산균만을 처리한 것(60%)에 비해 다소 증가함을 확인할 수 있었다.From Table 5, it was confirmed that the addition of Succinic acid or Soybean oil to the lactic acid bacteria slightly increased the algae activity to 62% and 64%, respectively (60%).
유산균에 Succinic acid와 Soybean oil을 모두 포함한 혼합물에서는 7일 후 살조활성이 현저히 증가하여 H2O2 보다 약간 낮은 76% 수준의 살조 활성을 나타내어 세가지 성분의 혼합으로 인한 시너지 효과가 있음을 시사하였다. In the mixture containing both Succinic acid and Soybean oil in lactic acid bacteria, the algal activity was significantly increased after 7 days, showing a slightly lower 76% level of algicide activity than H 2 O 2 , suggesting a synergistic effect due to mixing of the three components.
Claims (7)
Algae composition for controlling freshwater green algae containing a mixture of lactic acid bacteria, organic acids and fats and oils as an active ingredient.
상기 유산균은 락토바실러스 가사리, 락토바실러스 카세이, 락토바실러스 플란타럼, 루코노스톡멘센테로이드스, 락토바실러스 부크너리, 락토바실러스 파라카세이 및 락토바실러스 하비넨시스로 이루어진 군으로부터 선택된 하나 이상인 것을 특징으로 하는 담수 녹조 방제용 살조 조성물.
According to claim 1,
The lactic acid bacteria are characterized in that at least one selected from the group consisting of Lactobacillus gasari, Lactobacillus casei, Lactobacillus plantarum, Lucono stock mentheroids, Lactobacillus Buchnery, Lactobacillus paracasei, and Lactobacillus habinensis. Algae composition for controlling freshwater green algae.
상기 유산균은 락토바실러스 부크너리인 것을 특징으로 하는 담수 녹조 방제용 살조 조성물.
According to claim 2,
The lactic acid bacteria is a freshwater algae control algae composition, characterized in that the lactobacillus buknery.
상기 유기산은 숙신산, 시트르산 및 탄닌산으로 이루어진 군으로부터 선택된 하나 이상인 것을 특징으로 하는 담수 녹조 방제용 살조 조성물.
The method according to any one of claims 1 to 3,
The organic acid is a freshwater algae control composition for algae control, characterized in that at least one selected from the group consisting of succinic acid, citric acid and tannic acid.
상기 유기산의 농도는 0.5~3중량%인 것을 특징으로 하는 담수 녹조 방제용 살조 조성물.
The method of claim 4,
The concentration of the organic acid is 0.5 ~ 3% by weight of algae composition for controlling freshwater algae, characterized in that.
상기 유지는 식물성 유지인 것을 특징으로 하는 감수 녹조 방제용 살조 조성물.
The method according to any one of claims 1 to 3,
The fat or oil composition for controlling green algae according to the present invention, characterized in that it is a vegetable fat or oil.
상기 유지의 농도는 0.5~3중량%인 것을 특징으로 하는 담수 녹조 방제용 살조 조성물.The method of claim 6,
The concentration of the fat and oil is a freshwater algae control algae composition, characterized in that 0.5 to 3% by weight.
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