JP4041519B2 - Environmental purification method and environmental purification agent - Google Patents
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Description
本発明は、環境の浄化方法および当該方法に用いる環境浄化剤、並びにこれら浄化方法と環境浄化剤に使用できる細菌に関するものである。The present invention relates to an environmental purification method, an environmental purification agent used in the method, and bacteria that can be used in the purification method and the environmental purification agent.
近年、産業技術の発達や人工の増加に伴って、環境に対する影響が問題となっている。例えば、生活廃水や食品工場からの廃水などに含まれる窒素化合物やリン化合物は、河川や海において富栄養化の原因となり、赤潮などを引き起こす。また、屎尿や畜産汚水は、河川等を汚染するのみでなく、悪臭の原因にもなる。さらに、家庭などから出る生ゴミは、水分含量が多いことから焼却には多大なエネルギーを要するのみならず、悪臭を発するという問題がある。
上記環境問題のうち、生活廃水や工場などからの廃水の処理には、大きく分けて物理化学処理と生物処理とが用いられる。これら方法のうち、凝集,沈殿,濾過,吸着などの物理化学処理は処理スピードに優れる。その一方で、生物処理は、汚染物質を高濃度に含む廃水に対して処理効率が悪く、また、おしなべて時間がかかるといった問題はあるものの、穏和な条件で安価に実施でき、省エネルギーの観点からも優れていることから、生物処理方法が採用される場合が多い。
しかし、従来、リン化合物を分解できる細菌は見出されておらず、生物処理によっては、廃水中のリン化合物を低減することはできないと考えられていた。また、生物処理方法による廃水処理においては、汚染物質を効率的に処理できる細菌の探索が、大きなテーマの一つとなっている。
また、悪臭の発生も問題である。特に、下水設備がない地域での汲み取り式便所や、客席から近い場所に設置せざるを得ない列車内の便所では、悪臭の原因となるアミン系化合物や硫黄系化合物を速やかに低減する必要がある。
悪臭を低減する方法としては、原因物質を活性炭等の多孔体に吸着する方法や、化学的に分解する方法がある、しかし、多孔体の吸着能は飽和するため、その脱臭能力は限定的である。また、化学的な分解方法は、家庭での使用が危険な場合がある。そこで、細菌を利用して悪臭を低減する方法が、種々検討されている。
例えば特開平11−137953号公報には、微生物を担持する担体と、この担体に散水する散水設備と、散水した水を回収する貯水タンクであって硝化細菌を有するものとを備える脱臭装置が開示されている。しかし、散水設備や貯水タンクを設けるのは設備費用の点で問題があるのみでなく、列車内など限られたスペースには装置を設置することが困難である。この点に関して、特に脱臭作用等に優れる細菌を用いることによって、設備の問題の解決を図っている技術もある。
特開平5−153971号公報には、Bacillus megaterium種に属する細菌を使った悪臭発生抑制剤が記載されている。しかし、この文献の実施例に記載されている当該細菌の作用効果は、「悪臭の発生が抑えられた」といった定性的なものであり、実際にどの程度効果があるのか不明である。
一方、特開2001−70424号公報にはBacillus thuringiensis種などのBacillus属細菌を担持した生物脱臭剤の製造方法が開示されており、実施例にはアンモニア等の具体的な処理データも開示されている。しかし、添付されている図を参照すれば明らかな様に、当該技術はあくまで空気中の揮発性化合物を処理するものであって、悪臭の直接の原因である物質を処理するものではない。
さらに、特開平6−190028号公報と特開平10−248556号公報には、それぞれ、Clostriduim属とThermus属細菌を含む脱臭剤と、Pseudomonas属細菌を含む脱臭剤が開示されており、悪臭原因物質である硫化メチルと二硫化メチルを処理した実験例が記載されている。
また、身近な環境汚染例として、台所の排水口や排水溝,浴室の排水口,浴槽,便所の水洗用タンク,水槽などに発生する「ヌメリ」がある。このヌメリの正体は、主に雑菌やカビ、藻類が産生する多糖類である。従って、ヌメリの抑制には殺菌作用を有する化合物等が用いられる。しかし、殺菌剤では効果が緩慢である場合があり、また、家庭での使用は危険な場合がある。
ところで、Empedobacter brevis等のEmpedobacter属細菌は、不飽和脂肪酸に水酸基を付加する等の生物変換反応に用いられるものとして知られている(特開平15−144186号公報)。しかし、従来、Empedobacter属細菌を廃水処理などの環境浄化に用いた例はない。In recent years, the influence on the environment has become a problem with the development of industrial technology and the increase of artificial. For example, nitrogen compounds and phosphorus compounds contained in domestic wastewater and wastewater from food factories cause eutrophication in rivers and seas and cause red tides. Moreover, manure and livestock wastewater not only pollute rivers and the like, but also cause odors. Furthermore, the garbage generated from households has a problem that not only does it require a lot of energy for incineration due to its high water content, but it also produces a foul odor.
Among the environmental problems, physicochemical treatment and biological treatment are roughly divided into treatment of domestic wastewater and wastewater from factories. Of these methods, physicochemical treatments such as aggregation, precipitation, filtration and adsorption are excellent in processing speed. On the other hand, biological treatment is not efficient for wastewater containing a high concentration of pollutants, and generally takes time, but it can be carried out under mild conditions at a low cost. Because of its superiority, biological treatment methods are often adopted.
However, conventionally, no bacteria capable of decomposing phosphorus compounds have been found, and it has been thought that phosphorus compounds in wastewater cannot be reduced by biological treatment. In addition, in the wastewater treatment by the biological treatment method, searching for bacteria capable of efficiently treating pollutants is one of the major themes.
Also, the generation of malodor is a problem. In particular, it is necessary to quickly reduce amine-based compounds and sulfur-based compounds that cause foul odors in fetch-type toilets in areas where there are no sewage facilities and toilets in trains that must be installed close to passenger seats. is there.
As a method for reducing malodor, there are a method of adsorbing a causative substance to a porous body such as activated carbon and a method of chemically decomposing it. However, since the adsorption capacity of the porous body is saturated, its deodorizing ability is limited. is there. Also, chemical decomposition methods can be dangerous for home use. Accordingly, various methods for reducing malodor using bacteria have been studied.
For example, Japanese Patent Application Laid-Open No. 11-137553 discloses a deodorizing apparatus including a carrier for supporting microorganisms, a watering facility for watering the carrier, and a water storage tank for collecting the water sprayed and having nitrifying bacteria. Has been. However, providing a watering facility and a water storage tank not only has a problem in terms of facility cost, but it is difficult to install the device in a limited space such as in a train. In this regard, there is also a technology that attempts to solve the facility problem by using bacteria that are particularly excellent in deodorizing action and the like.
Japanese Patent Application Laid-Open No. 5-153971 describes a malodor generation inhibitor using bacteria belonging to the species Bacillus megaterium. However, the action and effect of the bacteria described in the examples of this document are qualitative such as “the occurrence of malodor is suppressed”, and it is unclear how much the effect is actually achieved.
On the other hand, Japanese Patent Application Laid-Open No. 2001-70424 discloses a method for producing a biodeodorant carrying Bacillus genus bacteria such as Bacillus thuringiensis species, and specific processing data such as ammonia is also disclosed in the examples. Yes. However, as is apparent with reference to the accompanying drawings, this technique only processes volatile compounds in the air, not a substance that directly causes malodor.
Furthermore, JP-A-6-190028 and JP-A-10-248556 disclose deodorizers containing Clostridium genus and Thermus genus bacteria, and deodorizers containing Pseudomonas genus bacteria, respectively, which cause malodor Experimental examples in which methyl sulfide and methyl disulfide are treated are described.
Examples of familiar environmental pollution include “numeric” that occurs in kitchen drains and drains, bathroom drains, bathtubs, toilet flush tanks, and water tanks. The true nature of this slime is a polysaccharide produced mainly by various germs, molds and algae. Therefore, a compound having a bactericidal action is used for suppressing slime. However, fungicides can be slow and can be dangerous to use at home.
By the way, Empedobacter genus bacteria such as Empedobacter brevis are known to be used for bioconversion reactions such as adding hydroxyl groups to unsaturated fatty acids (Japanese Patent Laid-Open No. 15-144186). However, there has been no example of using Empedobacter bacteria for environmental purification such as wastewater treatment.
上述したように、細菌を用いた廃水や悪臭の処理は、従来から行なわれていた。また、ヌメリを抑制する技術も知られている。しかし、有害な有機化合物や無機化合物、特に廃水CODの上昇の原因となる化合物や、悪臭の原因となるアミン系化合物や硫黄化合物、ヌメリの原因となる多糖類の生物処理については、より効率的な技術が求められている。特に、河川等の富栄養化の原因となるリン化合物を処理できる細菌は見出されていないことから、リン化合物の処理能力を有する細菌が切望されている。
そこで、本発明が解決すべき課題は、細菌により有害な有機化合物や無機化合物を処理する環境浄化方法、および当該方法に用いる環境浄化剤、並びにこれら環境浄化方法と環境浄化剤に用いることができる細菌を提供することにある。
本発明者らは、上記課題を解決すべく、環境浄化に有用な細菌を探索したところ、これまで環境浄化に用いられた例のないEmpedobacter属細菌が、廃水のCODや悪臭成分等のみならず、リン化合物濃度も低減できる能力を有することを見出して、本発明を完成した。
即ち、本発明の環境浄化方法は、Empedobacter属細菌および/またはその培養物を被処理物と接触させることを特徴とする。
また、本発明の環境浄化剤は、Empedobacter属細菌および/またはその培養物を含有することを特徴とする。
さらに、本発明に係る環境浄化方法と環境浄化剤で、特に優れた効果を発揮するEmpedobacter属細菌は、SIID2926−1b株(FERM BP−10360)である。As described above, wastewater and malodor treatment using bacteria has been conventionally performed. A technique for suppressing slime is also known. However, it is more efficient for biological treatment of harmful organic compounds and inorganic compounds, especially compounds that cause an increase in wastewater COD, amine compounds and sulfur compounds that cause malodor, and polysaccharides that cause slime. Technology is required. In particular, since no bacteria capable of treating phosphorus compounds that cause eutrophication of rivers and the like have been found, bacteria having the ability to treat phosphorus compounds are anxious.
Therefore, the problems to be solved by the present invention can be used in an environmental purification method for treating harmful organic compounds and inorganic compounds by bacteria, an environmental purification agent used in the method, and these environmental purification methods and environmental purification agents. To provide bacteria.
In order to solve the above-mentioned problems, the present inventors searched for bacteria useful for environmental purification. As a result, Empedobacter bacteria that have not been used for environmental purification so far are not only wastewater COD, malodorous components, etc. The present invention was completed by finding that it has the ability to reduce the phosphorus compound concentration.
That is, the environmental purification method of the present invention is characterized in that an Empedobacter bacterium and / or a culture thereof are brought into contact with an object to be treated.
In addition, the environmental purification agent of the present invention is characterized by containing an Empedobacter bacterium and / or a culture thereof.
Furthermore, the Empidobacter genus bacteria exhibiting particularly excellent effects with the environmental purification method and the environmental purification agent according to the present invention are SIID2926-1b strain (FERM BP-10360).
本発明に係る細菌であるEmpedobacter属細菌のうち、SIID2926−1b株は、下記の通り寄託機関に寄託されている。
(i) 寄託機関の名称およびあて名
名称: 独立行政法人産業技術総合研究所 特許生物寄託センター
あて名: 日本国 茨城県つくば市東1丁目1番地1 中央第6
(ii) 寄託日: 平成16年(2004年)6月30日
(iii) 寄託番号: FERM BP−10360
なお、SIID2926−1b株は、上記寄託機関に平成16年6月30日付で国内寄託された後(寄託番号:FERM P−20108)、本願出願前に国内寄託から国際寄託へ移管されたものである。
光学顕微鏡(オリンパス製,U−LH1000)を用いて観察した、SIID2926−1b株の細胞形態,グラム染色性,胞子の有無,鞭毛による運動性の有無、およびnutrient agar(Oxoid製)上30℃で24時間培養したコロニーの形態、更には、Barrow,G.I.とFeltham,R.K.A.,Cowan and Steel’s Manual for the Identification of Medical Bacteria. 3rd edition,Cambridge University Press.(1993年)に従ったカタラーゼ反応,オキシダーゼ反応およびブドウ糖の酸化/発酵(O/F)の試験結果は、表1に示す通りである。
SIID2926−1b株は、廃水のCOD,BOD,窒素化合物やリン化合物の低減と共に、悪臭の原因となるアミン系化合物や硫黄系化合物の発生抑制、ヌメリの原因となる多糖類の抑制について、優れた作用を有する。従って、SIID2926−1b株の他のEmpedobacter属細菌も、これら作用を有すると考えられ、SIID2926−1b株と同様に、環境浄化のために用いることができる。また、複数のEmpedobacter属細菌を併用して用いてもよい。
本発明の環境浄化方法は、Empedobacter属細菌および/またはその培養物を被処理物と接触させることを特徴とする。本発明方法によって、環境中における有害化合物、特に河川等の富栄養化の原因となる有機化合物,窒素化合物やリン化合物;悪臭の原因となるアミン系化合物や硫黄系化合物;ヌメリの原因となる多糖類;カビの発生を抑制することができる。
本発明方法で用いるEmpedobacter属細菌は、上記で説明した通りである。Empedobacter属細菌の培養物とは、Empedobacter属細菌の培養に用いた培地と、Empedobacter属細菌の生育に伴って当該培地に放出された化合物や酵素等を含むものをいう。Empedobacter属細菌自体が、有害化合物を分解できるが、細菌が産生する酵素等が同様の作用効果を有することも考えられる。
当該培地は、Empedobacter属細菌の培養に適するものであれば特に制限されない。例えば、純水,精製水,蒸留水などの水へ、糖類等の炭素栄養源、尿素やペプトン等の窒素栄養源、酵母エキス等の微量元素源などを適量添加したものを用いることができる。
本発明方法の被処理物は、有害な化合物を含むもの、或いは有害な化合物を発生させるものであれば、特に制限されない。例えば、生活廃水や食品工場廃水など河川等の富栄養化の原因となる廃水,悪臭源である屎尿や畜産汚水,生ゴミ等の食品残渣,微生物汚泥などを挙げることができる。また、悪臭が問題となるトイレ室内,便器,犬猫の糞尿場,灰皿,生ゴミ容器;ヌメリやカビが問題となる浴室の排水口や排水溝や壁,浴槽,便所の水洗用タンク,水槽,冷房機の室外機,生ゴミ容器などへ、Empedobacter属細菌および/またはその培養物を直接作用させてもよい。
本発明方法で用いる環境浄化剤は、Empedobacter属細菌および/またはその培養物を含有することを特徴とする。本発明の環境浄化剤によって、環境中における有害化合物、特に河川等の富栄養化の原因となる有機化合物,窒素化合物やリン化合物;悪臭の原因となるアミン系化合物や硫黄系化合物;ヌメリの原因となる多糖類を抑制することができる。従って、本発明の環境浄化剤は、廃水処理剤,悪臭抑制剤または脱臭剤,ヌメリ抑制剤または多糖類発生抑制剤,カビ発生抑制剤等として有用である。
本発明の環境浄化剤は、Empedobacter属細菌および/またはその培養物を被処理物へ直接投与するものであってもよいが、被処理物の種類に応じた形態としてもよい。
例えば、被処理物がトイレ室内や生ゴミ自体である場合には、Empedobacter属細菌および/またはその培養物を溶媒中に分散させ、これを被処理物へ散布してもよい。この場合に用いることができる溶媒は、水;エタノール等のアルコール;水とアルコールとの混合溶媒を挙げることができる。斯かる分散液の濃度は特に制限されないが、例えば、Empedobacter属細菌に対して溶媒を5000〜10000質量倍程度加えたものとすることができる。また、Empedobacter属細菌の作用効果を著しく減ずるものでない限り、菌床になり得るもの及びその組成物,pH調整剤,色素,消毒剤などを併用してもよい。なお、この分散液には、Empedobacter属細菌の培養液も含むものとする。
被処理物が廃水である場合には、浄化槽など従来の廃水処理設備へEmpedobacter属細菌および/またはその培養物を投入してもよい。その際には、培養物を投入する場合には特にその必要はないが、細菌自体を投与する場合には、炭素源,窒素源,微量元素源など、上記細菌の生育に適した栄養源を添加し、その他の条件も上記細菌の生育条件に即したものにする。また、栄養源として植物系の残渣を加えてもよい。植物系残渣は細菌の栄養源として優れるのみでなく、本来廃棄されるべきものの再利用を図ることができるからである。斯かる植物系残渣としては、例えば、米糠,おから,鋸くず,コーヒー粕,油粕,大豆粕,ヤシガラ,サトウキビバガス等を挙げることができ、本発明細菌に対しては、米糠とコーヒー粕が最適である。
上述した植物系残渣中で本発明細菌を培養したものは、そのまま廃水処理に用いることができるものとして有用である。その作成方法は、例えば、植物系残渣に本発明細菌と他の栄養源等を添加した後、十分に水分を与えて37〜39℃程度でインキュベートし、数日間発酵させる。これをそのまま被処理物に加えてもよいし、或いは一旦乾燥したものを被処理物に添加してもよい。
また、本発明の環境浄化剤は、Empedobacter属細菌および/またはその培養物を担体に担持したものであってもよい。使用できる担体としては、米糠,コーヒー粕,大豆粕,椰子ガラ,活性炭,シリカ多孔体,ガーゼ,不織布,布帛からなる群より選択される1または2以上を挙げることができる。この環境浄化剤は、例えば、担体にEmpedobacter属細菌の培養液を加えた上で、適度に乾燥することにより製造することができる。
本発明に係る環境浄化剤細菌の添加量は、例えば、廃水のCOD、悪臭の発生度合い、生ゴミなど固形成分の存在などに応じて適宜調整すればよい。被処理物が廃水である場合、一般的には、廃水1Lに対して菌体重量で0.3mg程度添加すればよい。被処理物がトイレ室内や生ゴミである場合には、Empedobacter属細菌および/またはその培養物の分散液である環境浄化剤を、悪臭が発生しない程度に適度散布すればよい。また、SIID2926−1b株は上述した通り30℃と37℃で生育するため、特に温度制御する必要はなく常温で処理してもよいが、寒冷地での実施など、場合によっては適温でインキュベートしてもよい。具体的な処理条件は、予備実験により決定すればよい。
以下、実施例を挙げて本発明をより具体的に説明するが、本発明はもとより下記実施例により制限を受けるものではなく、前・後記の趣旨に適合し得る範囲で適当に変更を加えて実施することも可能であり、それらはいずれも本発明の技術的範囲に含まれる。Among Empedobacter bacteria which are bacteria according to the present invention, SIID2926-1b strain is deposited with the depository as follows.
(I) Name and address of depositary institution Name: National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center Address: Japan 1-1-1 Higashi 1-chome Tsukuba City, Ibaraki Pref.
(Ii) Date of deposit: June 30, 2004 (iii) Deposit number: FERM BP-10360
The SIID2926-1b strain was deposited at the above depository in Japan on June 30, 2004 (deposit number: FERM P-20108) and was transferred from the domestic deposit to the international deposit before filing this application. is there.
Observed using an optical microscope (Olympus, U-LH1000), cell shape of SIID2926-1b strain, Gram staining, presence or absence of spores, presence or absence of motility by flagella, and on a neutral agar (Oxoid) at 30 ° C. Morphology of colonies cultured for 24 hours, as well as Barrow, G. et al. I. And Feltham, R .; K. A. , Cowan and Steel's Manual for the Identification of Medical Bacteria. 3rd edition, Cambridge University Press. The test results of catalase reaction, oxidase reaction and glucose oxidation / fermentation (O / F) according to (1993) are as shown in Table 1.
SIID2926-1b strain is excellent in reducing COD, BOD, nitrogen compounds and phosphorus compounds in wastewater, suppressing the generation of amine compounds and sulfur compounds that cause malodor, and suppressing polysaccharides causing slime. Has an effect. Accordingly, other Empedobacter bacteria belonging to the SIID2926-1b strain are also considered to have these actions, and can be used for environmental purification in the same manner as the SIID2926-1b strain. A plurality of Empedobacter bacteria may be used in combination.
The environmental purification method of the present invention is characterized by bringing an Empedobacter genus bacterium and / or a culture thereof into contact with an object to be treated. By the method of the present invention, harmful compounds in the environment, particularly organic compounds, nitrogen compounds and phosphorus compounds that cause eutrophication of rivers, etc .; amine compounds and sulfur compounds that cause malodors; Saccharides; generation of mold can be suppressed.
Empedobacter bacteria used in the method of the present invention are as described above. The culture of the genus Empedobacter refers to a culture medium used for culturing the bacterium of the genus Empedobacter and a compound, enzyme, or the like released into the medium along with the growth of the bacterium of the genus Empedobacter. Empedobacter genus bacteria themselves can decompose harmful compounds, but it is also conceivable that enzymes produced by bacteria have similar effects.
The medium is not particularly limited as long as it is suitable for culturing Empedobacter bacteria. For example, water obtained by adding an appropriate amount of a carbon nutrient source such as sugar, a nitrogen nutrient source such as urea or peptone, or a trace element source such as yeast extract to water such as pure water, purified water, or distilled water can be used.
The object to be treated of the method of the present invention is not particularly limited as long as it contains a harmful compound or generates a harmful compound. For example, wastewater that causes eutrophication of rivers and the like such as domestic wastewater and food factory wastewater, odor sources such as manure and livestock sewage, food residues such as raw garbage, microbial sludge, and the like. In addition, toilet rooms, toilets, dogs and cats' urine fields, ashtrays, garbage containers; bathroom drains, drains and walls, bathtubs, toilet flush tanks, water tanks where slime and mold are a problem , Empedobacter genus bacteria and / or cultures thereof may be directly applied to an outdoor unit of a cooling unit, a garbage container, or the like.
The environmental purification agent used in the method of the present invention is characterized by containing an Empedobacter genus bacterium and / or a culture thereof. By the environmental purifier of the present invention, harmful compounds in the environment, especially organic compounds, nitrogen compounds and phosphorus compounds which cause eutrophication of rivers, etc .; amine compounds and sulfur compounds which cause malodor; causes of slime Can be suppressed. Therefore, the environmental purification agent of the present invention is useful as a wastewater treatment agent, malodor inhibitor or deodorizer, slime inhibitor or polysaccharide occurrence inhibitor, mold occurrence inhibitor, and the like.
Although the environmental purification agent of this invention may administer the Empedobacter genus bacteria and / or its culture directly to a to-be-processed object, it is good also as a form according to the kind of to-be-processed object.
For example, when the object to be processed is a toilet room or garbage itself, Empedobacter genus bacteria and / or cultures thereof may be dispersed in a solvent and sprayed on the object to be processed. Examples of the solvent that can be used in this case include water; an alcohol such as ethanol; and a mixed solvent of water and alcohol. The concentration of such a dispersion is not particularly limited. For example, the dispersion may be about 5000 to 10000 mass times added to Empedobacter bacteria. In addition, as long as the effects of Empedobacter bacteria are not significantly reduced, those capable of forming a fungus bed, compositions thereof, pH adjusters, pigments, disinfectants, and the like may be used in combination. In addition, this dispersion liquid shall also contain the culture solution of Empedobacter genus bacteria.
When the material to be treated is wastewater, Empedobacter bacteria and / or cultures thereof may be put into a conventional wastewater treatment facility such as a septic tank. In that case, when the culture is introduced, it is not particularly necessary. However, when the bacterium itself is administered, a nutrient source suitable for the growth of the bacterium, such as a carbon source, a nitrogen source, and a trace element source. The other conditions should be adjusted to the growth conditions of the bacteria. Moreover, you may add a plant-type residue as a nutrient source. This is because plant residue is not only excellent as a nutrient source for bacteria, but can also be reused for what should be discarded. Examples of such plant residues include rice bran, okara, sawdust, coffee lees, oil lees, soybean lees, coconut husks, sugarcane bagasse and the like. Is optimal.
What culture | cultivated this invention bacteria in the plant-type residue mentioned above is useful as what can be used for wastewater treatment as it is. The preparation method includes, for example, adding the bacterium of the present invention and other nutrients to the plant residue, sufficiently injecting water, incubating at about 37 to 39 ° C., and fermenting for several days. This may be added as it is to the object to be processed, or once dried, it may be added to the object to be processed.
Moreover, the environmental purification agent of this invention may carry | support the Empedobacter genus bacteria and / or its culture | cultivation on the support | carrier. Examples of the carrier that can be used include one or more selected from the group consisting of rice bran, coffee koji, soybean koji, coconut shell, activated carbon, porous silica, gauze, non-woven fabric, and fabric. This environmental purification agent can be produced, for example, by adding a culture solution of Empedobacter bacteria to a carrier and drying it appropriately.
What is necessary is just to adjust suitably the addition amount of the environmental purification | cleaning agent bacteria which concern on this invention according to the COD of wastewater, the generation | occurrence | production degree of malodor, presence of solid components, such as garbage. When the object to be treated is waste water, generally, it is sufficient to add about 0.3 mg in terms of bacterial cell weight to 1 L of waste water. In the case where the object to be treated is a toilet room or garbage, an environmental purification agent which is a dispersion of Empedobacter bacteria and / or cultures thereof may be appropriately sprayed to such an extent that no bad odor is generated. In addition, since the SIID2926-1b strain grows at 30 ° C. and 37 ° C. as described above, it is not necessary to control the temperature in particular, and it may be treated at room temperature, but it may be incubated at an appropriate temperature in some cases such as in a cold region. May be. Specific processing conditions may be determined by preliminary experiments.
EXAMPLES Hereinafter, the present invention will be described more specifically with reference to examples. However, the present invention is not limited by the following examples, but may be appropriately modified within a range that can meet the purpose described above and below. It is also possible to implement, and they are all included in the technical scope of the present invention.
実施例1 16SrDNA塩基配列解析
SIID2926−1b株(FERM BP−10360)の菌体について、ゲノムDNAを抽出した。ゲノムDNAの抽出には、Insta Gene Matrix(BIO RAD製)を使用し、プロトコールはBIO RAD社のものに従った。抽出したゲノムDNAを鋳型として、PCRにより16SrDNAのうち5’末端側約500bpの領域を増幅した。その後、増幅された塩基配列をシーケンシングし、塩基配列を決定した。PCRとサイクルシークエンスにはMicro Seq 500 16SrDNA Bacterial Identification PCR Kit(Applied Biosystems製)を、サーマルサイクラーにはGene Amp PCR System 9600(Applied Biosystems製)を、DNAシーケンサーにはABI PRISM 3100 DNA Sequencer(Applied Biosystems製)を使用した。また、PCRからサイクロシークエンスまでの基本的操作は、Applied Biosystems社のプロトコール(P/N4346296 Rev.B)に従った。
次に、得られた塩基配列情報を用いて、属種の同定を行なった。先ず、各属種の基準株のデータベースであるMicro Seq Bacterial 500 Library v.0023(Applied Biosystems)を用い、解析ソフトウェアとしてMicro Seq Microbial Identification System Software V.1.4.1を、アルゴリズムとしてBLASTを使用して、既知細菌との相同率を求めた。相同率上位5位までの結果を表3に示す。
また、念のために国際塩基配列データベース(GeneBank/DDBJ/EMBL)による同定も行なった。相同率上位5位までの結果を表4に示す。
以上の結果から、SIID2926−1b株はEmpedobacter属とFlavobacterium属への近縁性が示唆されるものの、これら属に分類される基準株への相同性は認められなかったことから属種レベルで全く新規の細菌であり、これら属が含まれるFlavobacteriaceae科に分類されるものであると結論付けた。
実施例2 脂肪酸組成分析
SIID2926−1b株をTripticase Soy(SCD)Agar(Becton Dickinson製)平板培地に植菌し、30℃で24時間培養した。この平板4枚分の菌体をセルスクレイパーでかき取り、13mL容のガラスバイアルの底部に擦り付けた。
この菌体に鹸化液(水酸化ナトリウム45g,メタノール150mL,蒸留水150mLを均一混合したもの)1mLを加え、100℃のウォーターバスによってガラスバイアルを恒温状態で静置した。反応開始5分後に一旦ウォーターバスから取り出し、軽く攪拌してから戻し、菌体の鹸化を更に30分間行なった。鹸化した試料液中にメチル化試薬(6.0M塩酸325mLとメタノール275mLを均一混合したもの)2mLを加え、80℃に設定したウォーターバス中で10分間反応させ、鹸化により生じた脂肪酸をメチルエステル化した。
試料液中に抽出液(ヘキサン200mLとt−ブチルメチルエーテル200mLとの混合溶液)1.25mLを加え、シェーカーで10分間攪拌した後静置した。二層分離した水層をパスツールピペットで除去した。残った有機層に洗浄液(水酸化ナトリウム10.8gと蒸留水900mLを均一混合したもの)3mLを加えてシェーカーで5分間攪拌したところ、エマルジョン状態になったため、飽和食塩水を数滴滴下することによって二層分離させた。分離した有機層の上部2/3をパスツールピペットで取り出し、これをガスクロマトグラフィー用サンプルバイアルに移し、測定用試料とした。
上記試料について、ガスクロマトグラフィーシステム HP6890(Hewlett−Packard製)を用いて脂肪酸組成を測定した。結果を図1に示す。また、この結果について、Sherlock Microbial Identification Systemを用いて、既知菌種との相同性検索を行なった。結果を表5に示す。
当該結果より、SIID2926−1b株は、Flavobacteriaceae科に分類されるEmpedobacter brevisとの近縁性が示唆されるものの、既知の菌種とは一致しないものであることが明らかとなった。
実施例3 分子系統樹
上記実施例1で得られた16SrDNA塩基配列解析の結果から推定されたSIID2926−1b株の近縁菌群に含まれる基準菌株について、データベース(GenBank/DDBJ/EMBL)から16SrDNA塩基配列の上流側を取得し、分子系統樹を作成した。その際、アラインメント(塩基配列間の対応が最も良くなる様に配列データへ適当なギャップを挿入する処理)には、アラインメントソフトであるClustal X(Thompson,J.D.ら,「The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools」,Nucleic Acids Reserach,24,pp.4876−4882(1997))を用い、分子系統樹の作成には、コンピューターソフトウェアであるMEGA 2(Kumar,R.ら,「MEGA 2: Molecular Evolutionary Genetics Analysis software」,Bioinformatics,17,pp.1244−1245(2001))を使用した。なお、分子系統樹の推定には近隣結合法(Saito,N.ら,「The neighbor−joining method: a new method for reconstructing phylogenetic trees」,Mol.Biol.Evol,4,pp.406−425(1987))を用い、樹形の妥当性を示すブートストラップ(Felsenstein,J.,「Confidence limits on phylogenies:an approach using the bootstrap.」,Evolution,39,pp.783−791(1985))は1000回発生させた。また、SIID2926−1b株を含むクラスターは、ブートストラップ値99%と100%で支持された。結果を図2に示す。
当該結果より、SIID2926−1b株の16SrDNA塩基配列は、Empedobacter brevisの16SrDNA塩基配列が形成するクラスターに含まれることが分かる。従って、上記実施例1と2の結果も合わせて考察すると、SIID2926−1b株は、Empedobacter属に分類される新規な菌株であると結論付けることができる。
実施例4 脱臭能力試験
先ず、米糠にSIID2926−1b株とコーヒー粕と尿素とを加え、地下水を吹き付けたものを37〜39℃で5日間インキュベートし、次いで適度に自然乾燥することによって、SIID2926−1b株を含む植物系残渣を作成した。次に、12L容の密閉真空デシケーターに屎尿処理場原水10Lを取り、この原水1L当たり10gの割合(合計100g)で当該植物系残渣を添加した。このデシケーターを密閉し、震蕩しつつ25℃でインキュベートした。所定時間毎に、シリンジにより気相部分の空気を抜出し、JIS K0099の規定に基づいてインドフェノール吸光度法によって、気相部分のアンモニア濃度(v/v ppm)を測定した。結果を表6に示す。
実施例5 廃水処理試験
先ず、グリストラップで処理された後に排出されている食品(かまぼこ)製造工場由来の廃水(放流量:25m3/日)について、pH、化学的酸素要求量、生物化学的酸素要求量、浮遊物質量、全窒素含有量、全リン含有量およびn−ヘキサン抽出物質量(鉱油類と動植物油脂類)を測定した。
次に、当該グリストラップに、上記実施例4で用いたSIID2926−1b株を含む植物系残渣を1日当たり500g加えて攪拌する処理を20日間連続して行なった。その後における廃水について、同様の条件でpH等を測定した。結果を表7に示す。
実施例6 廃水処理試験
先ず、機械部品製造工場に設置されている浄化槽(350人槽)の流入水と放流水について、pH、化学的酸素要求量、生物化学的酸素要求量、浮遊物質量、窒素化合物含有量およびリン化合物含有量を、表7に示す方法により測定した。この浄化槽は、食堂からの生活廃水と屎尿を合わせた廃水を1日当たり46m3処理しているものであり、通常の活性汚泥処理方式を採用したものである。しかし、当該浄化槽の汚泥負荷と流量負荷を過小設定したことによると推測される、放流水中のCODとBOD等の浄化不十分などのトラブルが頻発していた。
上記測定後、当該浄化槽へ上記実施例4で用いたSIID2926−1b株を含む植物系残渣を50kg投入し、投入4日後において、同様の測定を行なった。さらに、最初の投入から1週間ごとに植物系残渣を4kgずつ計16kg投入し、最後の投入を行なった後で再度同様の測定を行なった。結果を表8に示す。
実施例7 悪臭抑制試験
水4000質量部に、SIID2926−1b株0.5質量部、グルコース10質量部、ペプトン5質量部および酵母エキス2.5質量部を添加し、30℃で5日間培養した。当該培養液5mLを水500mLに添加した。これを環境浄化剤Aとする。当該環境浄化剤Aを、猫の糞尿場所である砂場に10日間にわたり毎朝8時に約2〜3mL散布し、悪臭の発生状態を、毎朝7時に3人でチェックした上での合意により判定した。環境浄化剤Aの散布前と散布後における悪臭の発生状態を、悪臭がある場合を×、悪臭が少ない場合を△、悪臭がない場合を○として表9に示す。
実施例8 悪臭抑制試験
上記実施例7の環境浄化剤Aを、家族3人が使用する汲み取り式便所のタンクへ、20日間にわたり毎朝8時に約15〜20mL投入し、便器蓋を除いた際における悪臭の発生状態を、毎朝7時に3人でチェックした上での合意により判定した。環境浄化剤Aの散布前と散布後における悪臭の発生状態を、悪臭がある場合を×、悪臭が少ない場合を△、悪臭がない場合を○として表10に示す。
実施例9 悪臭抑制試験
上記実施例7で調製した培養液4000質量部に、米糠10000質量部、コーヒー粕1000質量部、尿素100質量部、蜂蜜2質量部、砂糖2質量部を添加した。培養に伴う発熱による昇温に注意し、55℃を超えない様に時々攪拌しつつ、5日間静置培養した。その後、培養液を自然乾燥してSIID2926−1b株を仮死状態とし、環境浄化剤Bを得た。当該環境浄化剤Bの水分を米穀水分計により測定したところ、13%だった。
この環境浄化剤Bを、上記実施例8と同様の方法により便所タンクへ約20〜25g投入し、悪臭抑制作用を調べた。環境浄化剤Bの散布前と散布後における悪臭の発生状態を、悪臭がある場合を×、悪臭が少ない場合を△、悪臭がない場合を○として結果を表11に示す。
実施例10 悪臭抑制試験
上記実施例9で得た環境浄化剤Bを、家族3人の台所生ゴミを一時保存する容量100Lの蓋付ポリバケツへ、30日間にわたり毎朝8時に約8〜10g散布して攪拌し、毎日朝、昼、晩の3食分の生ゴミをその上に蓄積した。蓋を除いた際における悪臭の発生状態を、毎朝7時に3人でチェックした上での合意により判定した。環境浄化剤Bの散布前と散布後における悪臭の発生状態を、悪臭がある場合を×、悪臭が少ない場合を△、悪臭がない場合を○として表12に示す。
実施例11 悪臭抑制試験
上記実施例7の環境浄化剤Aを、自家用車の灰皿に、20日間にわたり毎晩20時に約5〜7mL散布し、悪臭の発生状態を、毎朝7時に3人でチェックした上での合意により判定した。環境浄化剤Aの散布前と散布後における悪臭の発生状態を、悪臭がある場合を×、悪臭が少ない場合を△、悪臭がない場合を○として表13に示す。
実施例12 カビおよびヌメリの発生抑制試験
上記実施例7で調製した培養液10質量部を、水1000質量部に加えて分散混合した。これを環境浄化剤Cとした。当該環境浄化剤Cを、いったん洗浄した浴室の壁と浴槽へ、60日にわたり1日当たり3〜5mL散布した。60日後の実験後において、浴室壁にも浴槽にも、カビやヌメリは全く見られなかった。従って、本発明に係る環境浄化剤のカビおよびヌメリの発生抑制作用が実証された。
実施例13 カビおよびヌメリの発生抑制試験
上記実施例12の環境浄化剤Cを、60日にわたって3日に1回3〜5mLずつ便所の水洗用タンクへ添加した。60日後の実験後において、水洗用タンクにも便器にも、カビやヌメリは全く見られなかった。従って、本発明に係る環境浄化剤のカビおよびヌメリの発生抑制作用が証明された。
実施例14 ヌメリ抑制試験
上記実施例9で得た環境浄化剤B50gを、ポリエステル繊維不織布の袋に入れて袋口部を封じ、これを家族3人が使用する台所の排水口のプラスチック製カゴに入れた。3ヵ月後にわたって、上記プラスチック製カゴのヌメリの発生状態を、3人でチェックした上での合意により毎日判定したところ、一度もヌメリは発生しなかった。従って、本発明に係る環境浄化剤のヌメリ抑制作用が証明された。
実施例15 ヌメリ抑制試験
上記実施例7で調製した培養液10000質量部に多孔性炭(平均粒子径:2〜5mm)1000質量部を浸漬し、30℃で時々攪拌しつつ5日間静置した。その後、多孔性炭のみを取り出し、自然乾燥することによって、水分12%の環境浄化剤Dを得た。当該環境浄化剤Dをポリエステル繊維不織布の袋に150g入れて袋口部を封じ、24時間風呂のフィルター部に合わせて成形し、装着して使用した。3ヵ月後にわたって、フィルター部におけるヌメリの発生状態を、毎朝7時に3人でチェックした上での合意により毎日判定したところ、一度もヌメリは発生しなかった。従って、本発明に係る環境浄化剤のヌメリ抑制作用が証明された。
実施例16 ヌメリ抑制試験
上記実施例7で調製した培養液10000質量部に活性炭素繊維(平均繊維径:100μm,平均繊維長:50mm)300質量部を浸漬し、32℃で時々攪拌しつつ5日間静置した。その後、活性炭素繊維のみを取り出し、自然乾燥することによって、水分14%の環境浄化剤Eを得た。当該環境浄化剤Eをポリエステル繊維不織布の袋に150g入れて袋口部を封じ、上記実施例17と同様に24時間風呂のフィルター部に装着して使用した。2ヵ月後にわたって、フィルター部におけるヌメリの発生状態を、毎朝7時に3人でチェックした上での合意により毎日判定したところ、一度もヌメリは発生しなかった。従って、本発明に係る環境浄化剤のヌメリ抑制作用が証明された。
実施例17 有機化合物分解試験
上記実施例9で得た環境浄化剤Bを、ペットの犬の糞尿を蓄積している土中の容器に、1ヵ月にわたり毎日1回30g程度を投入し、同時に水を適量散布し、3〜5日に1回攪拌した。その間、容器は通常通り使用し、犬の糞尿を蓄積した。また、比較のために、別途、環境浄化剤を用いない以外は同様の試験を行なった。糞尿は逐一半分とし、実施例と比較例へそれぞれ等量ずつ蓄積した。
試験開始後1ヵ月経過後、本発明の環境浄化剤Bを用いた場合には、糞尿の蓄積増加がほとんどない上に、悪臭の発生も極めて少なかった。一方、環境浄化剤Bを用いなかった場合は、糞尿の蓄積は数倍多く、悪臭も発生した。従って、本発明に係る環境浄化剤は、有害な有機化合物を分解できることが実証された。
実施例18 有機化合物分解試験
上記実施例9で得た環境浄化剤Bを、台所の生ゴミを蓄積している土中の容器に、1ヵ月にわたり毎日1回生ゴミと共に25g程度を投入し、同時に水を適量散布し、3〜5日に1回攪拌した。また、比較のために、別途、環境浄化剤を用いない以外は同様の試験を行なった。生ゴミは1日分を蓄積しておき、それを半分にし、実施例と比較例へそれぞれ等量ずつ蓄積した。
試験開始後1ヵ月経過後、本発明の環境浄化剤Bを用いた場合には、生ゴミの蓄積増加がほとんどない上に、悪臭の発生も極めて少なかった。一方、環境浄化剤Bを用いなかった場合は、生ゴミの蓄積は数倍多く、悪臭も発生した。従って、本発明に係る環境浄化剤は、有害な有機化合物を分解できることが実証された。Example 1 16S rDNA base sequence analysis Genomic DNA was extracted from the cells of SIID2926-1b strain (FERM BP-10360). For extraction of genomic DNA, Insta Gene Matrix (manufactured by BIO RAD) was used, and the protocol followed that of BIO RAD. Using the extracted genomic DNA as a template, a region of about 500 bp on the 5 ′ end side of 16SrDNA was amplified by PCR. Thereafter, the amplified base sequence was sequenced to determine the base sequence. For PCR and cycle sequencing, Micro Seq 500 16SrDNA Bacterial Identification PCR Kit (Applied Biosystems) is used, and for thermal cyclers, Gene Amp PCR System 9100 (Applied Biosystems is available). )It was used. In addition, the basic operation from PCR to cyclosequence followed the protocol of Applied Biosystems (P / N 4346296 Rev. B).
Next, genus species were identified using the obtained base sequence information. First, Micro Seq Bacterial 500 Library v. Which is a database of reference strains of each genus species. 0023 (Applied Biosystems) using Micro Seq Microbial Identification System Software V. as analysis software. Using BLAST as the algorithm for 1.4.1, the homology rate with known bacteria was determined. Table 3 shows the results of the top 5 homology ratios.
Moreover, the identification by the international base sequence database (GeneBank / DDBJ / EMBL) was also performed just in case. Table 4 shows the results of the top 5 homology rates.
Although the above results suggest that the SIID2926-1b strain is closely related to the genus Empedobacter and the Flavobacterium genus, homology to the reference strains classified into these genera was not recognized. It was concluded that this is a new bacterium and is classified into the Flavobacteriumaceae family that includes these genera.
Example 2 Fatty Acid Composition Analysis The SIID2926-1b strain was inoculated on a Tripticase Soy (SCD) Agar (manufactured by Becton Dickinson) plate culture and cultured at 30 ° C. for 24 hours. The cells of 4 plates were scraped with a cell scraper and rubbed against the bottom of a 13 mL glass vial.
1 mL of a saponification solution (sodium hydroxide 45 g, methanol 150 mL, distilled water 150 mL uniformly mixed) was added to the cells, and the glass vial was allowed to stand at a constant temperature in a 100 ° C. water bath. After 5 minutes from the start of the reaction, it was once taken out of the water bath, gently stirred and then returned, and the bacterial cells were further saponified for 30 minutes. To the saponified sample solution, add 2 mL of a methylating reagent (uniformly mixed 325 mL of 6.0 M hydrochloric acid and 275 mL of methanol) and react for 10 minutes in a water bath set at 80 ° C. Turned into.
1.25 mL of an extract (a mixed solution of 200 mL of hexane and 200 mL of t-butyl methyl ether) was added to the sample solution, and the mixture was stirred for 10 minutes with a shaker and allowed to stand. The aqueous layer separated into two layers was removed with a Pasteur pipette. To the remaining organic layer was added 3 mL of a washing solution (uniformly mixed 10.8 g of sodium hydroxide and 900 mL of distilled water) and stirred for 5 minutes with a shaker. As a result, the emulsion became emulsion. Drop a few drops of saturated saline. Separated into two layers. The upper 2/3 of the separated organic layer was taken out with a Pasteur pipette, transferred to a gas chromatography sample vial, and used as a measurement sample.
About the said sample, the fatty acid composition was measured using gas chromatography system HP6890 (made by Hewlett-Packard). The results are shown in FIG. Moreover, the homology search with the known microbial species was performed about this result using the Sherlock Microbial Identification System. The results are shown in Table 5.
From these results, it was clarified that the strain SIID2926-1b is closely related to Empedobacter brevis classified into the Flavobacteriaceae family, but is not consistent with known bacterial species.
Example 3 Molecular Phylogenetic Tree Regarding the reference strains included in the related strains of SIID2926-1b strain estimated from the results of the 16S rDNA base sequence analysis obtained in Example 1 above, 16SrDNA from the database (GenBank / DDBJ / EMBL) The upstream side of the base sequence was obtained and a molecular phylogenetic tree was created. At that time, alignment (processing for inserting an appropriate gap into the sequence data so that the correspondence between the base sequences is the best) is Clustal X (Thompson, JD et al., “The Clustal X Windows”, which is alignment software). interface: flexible Strategies for multiple sequence alignment aided by quality analysis tools, Nucleic Acids Research, 24, pp. 4876-4882 (1997). R. et al., “MEGA 2: Molecular Evolutionary Genetics Analysis. is software ", Bioinformatics, 17, was used pp.1244-1245 (2001)). For estimation of the molecular phylogenetic tree, the neighbor joining method (Saito, N. et al., “The new method for regenerating phylogenetic trees”, Mol. Biol. Evol, 4, pp. 406-4425 (pp. 406-4425). )), And the bootstrap (Felsenstain, J., “Confidence limits on phylogenies: an approach using the bootstrap.”, Evolution, 39, pp. 783-791 (1985)) is 1000 times. Generated. In addition, the cluster containing SIID2926-1b strain was supported with bootstrap values of 99% and 100%. The results are shown in FIG.
From the results, it can be seen that the 16S rDNA base sequence of SIID2926-1b strain is included in the cluster formed by the 16S rDNA base sequence of Empedobacterium brevis. Therefore, considering the results of Examples 1 and 2 together, it can be concluded that the SIID2926-1b strain is a novel strain classified into the genus Empedobacter.
Example 4 Deodorizing ability test First, SID2926-1b strain, coffee lees and urea were added to rice bran, and groundwater sprayed was incubated at 37-39 ° C. for 5 days, and then naturally dried to an appropriate extent. A plant residue containing strain 1b was prepared. Next, 10 L of raw wastewater treatment plant raw water was taken in a 12 L sealed vacuum desiccator, and the plant residue was added at a rate of 10 g per 1 L of the raw water (total of 100 g). The desiccator was sealed and incubated at 25 ° C. with shaking. At predetermined intervals, the air in the gas phase portion was extracted with a syringe, and the ammonia concentration (v / v ppm) in the gas phase portion was measured by the indophenol absorbance method based on the provisions of JIS K0099. The results are shown in Table 6.
Example 5 Wastewater treatment test First, pH, chemical oxygen demand, biochemical quality of wastewater (discharge flow rate: 25 m 3 / day) derived from a food (kamaboko) manufacturing plant discharged after being treated with grease wrap. The oxygen demand, the amount of suspended solids, the total nitrogen content, the total phosphorus content and the amount of n-hexane extract (mineral oils and animal and vegetable fats and oils) were measured.
Next, the plant residue containing the SIID2926-1b strain used in Example 4 was added to the grease trap and stirred for 20 days. About the waste water after that, pH etc. were measured on the same conditions. The results are shown in Table 7.
Example 6 Wastewater treatment test First, regarding the influent and effluent water of the septic tank (350 person tank) installed in the machine component manufacturing factory, pH, chemical oxygen demand, biochemical oxygen demand, suspended solids, The nitrogen compound content and the phosphorus compound content were measured by the methods shown in Table 7. The septic tank is intended that wastewater per day 46m 3 processing the combined domestic wastewater and sewage from canteen, is obtained by employing the conventional activated sludge treatment method. However, troubles such as inadequate purification of COD, BOD, etc. in the discharged water, which are presumed to be caused by setting the sludge load and flow rate load of the septic tank too small, occurred frequently.
After the measurement, 50 kg of the plant residue containing the SIID2926-1b strain used in Example 4 was introduced into the septic tank, and the same measurement was performed 4 days after the introduction. Furthermore, a total of 16 kg of plant-based residue was added every 4 weeks from the first input, and the same measurement was performed again after the final input. The results are shown in Table 8.
Example 7 Malodor Control Test To 4000 parts by mass of water, 0.5 part by mass of SIID2926-1b strain, 10 parts by mass of glucose, 5 parts by mass of peptone and 2.5 parts by mass of yeast extract were added and cultured at 30 ° C. for 5 days. . 5 mL of the culture solution was added to 500 mL of water. This is designated as environmental purification agent A. About 2 to 3 mL of the environmental purification agent A was applied to a sandbox which is a cat excrement place for 10 days every morning at 8 o'clock in the morning, and the state of occurrence of bad odor was determined by an agreement after checking by 3 people every morning at 7 o'clock. Table 9 shows the state of occurrence of malodor before and after the application of the environmental purification agent A as x when there is malodor, Δ when there is little malodor, and ○ when there is no malodor.
Example 8 Malodor Control Test When the environmental purification agent A of Example 7 was put into a tank of a draw-out toilet used by three families for about 15 to 20 mL every morning for 8 days at 8 o'clock and the toilet lid was removed The state of occurrence of bad odor was judged by agreement after checking by three people at 7 am every morning. Table 10 shows the occurrence of malodor before and after the application of the environmental purification agent A as x when there is malodor, Δ when there is little malodor, and ○ when there is no malodor.
Example 9 Malodor Control Test To 4000 parts by mass of the culture solution prepared in Example 7 above, 10,000 parts by mass of rice bran, 1000 parts by mass of coffee koji, 100 parts by mass of urea, 2 parts by mass of honey, and 2 parts by mass of sugar were added. Paying attention to the temperature rise due to the heat generated by the culture, the culture was allowed to stand for 5 days with occasional agitation so as not to exceed 55 ° C. Thereafter, the culture broth was naturally dried to make the SIID2926-1b strain as a dead state, and an environmental purification agent B was obtained. It was 13% when the water | moisture content of the said environmental purification agent B was measured with the rice grain moisture meter.
About 20 to 25 g of this environmental purification agent B was put into a toilet tank in the same manner as in Example 8 above, and the malodor control action was examined. Table 11 shows the results of the occurrence of bad odor before and after the application of the environmental purification agent B, where x is when there is a bad odor, Δ when there is little bad odor, and ○ when there is no bad odor.
Example 10 Malodor Control Test About 8 to 10 g of the environmental purification agent B obtained in Example 9 above was applied to a 100-liter lidded plastic bucket for temporarily storing kitchen garbage from three family members every morning at 8 am for 30 days. Each day, three meals of morning, noon and evening were accumulated on top of it. The state of occurrence of bad odor when the lid was removed was judged by an agreement after three people checked at 7 am every morning. Table 12 shows the generation state of malodor before and after the application of the environmental purification agent B as x when there is malodor, Δ when there is little malodor, and circle when there is no malodor.
Example 11 Odor Suppression Test The environmental purification agent A of Example 7 was sprayed on an ashtray of a private car for about 5 to 7 mL every night for 20 days at 20 o'clock, and the occurrence of malodor was checked by three people at 7 o'clock every morning. Judged by agreement above. Table 13 shows the occurrence of malodor before and after spraying of the environmental purification agent A, where x is when there is malodor, Δ when there is little malodor, and ○ when there is no malodor.
Example 12 Mold and slime generation suppression test 10 parts by mass of the culture solution prepared in Example 7 above was added to 1000 parts by mass of water and dispersed and mixed. This was designated as environmental purifier C. 3 to 5 mL of the environmental purification agent C per day was sprayed over the bathroom walls and bathtubs once washed. After the
Example 13 Generation | occurrence | production suppression test of mold | fungi and slime The environmental purification agent C of the said Example 12 was added to the flush tank of the toilet once every 3 days over 60 days. After the experiment after 60 days, neither mold nor slime was found in the washing tank or the toilet bowl. Therefore, it was proved that the environmental purifier according to the present invention suppresses the occurrence of mold and slime.
Example 14 Numerous Suppression Test 50 g of the environmental purification agent B obtained in Example 9 above is placed in a polyester fiber non-woven bag, the bag mouth is sealed, and this is placed in a plastic basket at the kitchen drain used by three family members. I put it in. Three months later, the occurrence of slime in the plastic basket was checked daily by an agreement after checking by three people, and no slime occurred. Therefore, the slime suppression effect of the environmental cleaner according to the present invention was proved.
Example 15 Numerous Suppression Test 1000 parts by mass of porous charcoal (average particle size: 2 to 5 mm) was immersed in 10000 parts by mass of the culture solution prepared in Example 7 and allowed to stand for 5 days with occasional stirring at 30 ° C. . Thereafter, only the porous charcoal was taken out and naturally dried to obtain an environmental purification agent D having a moisture content of 12%. 150 g of the environmental purification agent D was put in a bag of non-woven polyester fiber, the bag mouth was sealed, molded according to the filter part of the bath for 24 hours, mounted and used. Three months later, the occurrence of slime in the filter section was judged every day by an agreement after checking by three people at 7 am every morning, and no slime occurred. Therefore, the slime suppression effect of the environmental cleaner according to the present invention was proved.
Example 16 Numerity Inhibition Test 300 parts by mass of activated carbon fiber (average fiber diameter: 100 μm, average fiber length: 50 mm) was immersed in 10000 parts by mass of the culture solution prepared in Example 7 above. Let stand for days. Thereafter, only the activated carbon fiber was taken out and naturally dried to obtain an environmental purification agent E having a moisture content of 14%. 150 g of the environmental purification agent E was put in a bag of non-woven polyester fiber, the bag mouth was sealed, and it was used by being mounted on a filter part of a bath for 24 hours in the same manner as in Example 17. Two months later, the occurrence of slime in the filter section was judged every day by an agreement after checking by three people at 7 am every morning, and no slime occurred. Therefore, the slime suppression effect of the environmental cleaner according to the present invention was proved.
Example 17 Decomposition test of organic compound About 30 g of the environmental purification agent B obtained in the above Example 9 was poured into a soil container storing pet dog urine once a day for one month, Was sprayed in an appropriate amount and stirred once every 3 to 5 days. Meanwhile, the container was used as usual and accumulated dog manure. For comparison, a similar test was conducted except that no environmental cleaner was used. Feces and urine were halved and accumulated in equal amounts in the examples and comparative examples.
One month after the start of the test, when the environmental purification agent B of the present invention was used, there was almost no increase in the accumulation of feces and urine, and the generation of malodor was extremely small. On the other hand, when the environmental purification agent B was not used, feces and urine accumulated several times more, and malodors were also generated. Therefore, it was demonstrated that the environmental purification agent according to the present invention can decompose harmful organic compounds.
Example 18 Organic Compound Decomposition Test About 25 g of the environmental cleaning agent B obtained in Example 9 above was put into a soil container storing kitchen garbage once a day for one month together with garbage. An appropriate amount of water was sprayed and stirred once every 3 to 5 days. For comparison, a similar test was conducted except that no environmental cleaner was used. One day's worth of raw garbage was accumulated, halved, and an equal amount was accumulated in each of the examples and comparative examples.
One month after the start of the test, when the environmental purification agent B of the present invention was used, there was almost no increase in the accumulation of raw garbage, and there was very little malodor generation. On the other hand, when the environmental purification agent B was not used, the accumulation of garbage was several times greater and a bad odor was also generated. Therefore, it was demonstrated that the environmental purification agent according to the present invention can decompose harmful organic compounds.
本発明の細菌は、有害な化合物に対して優れた処理能力を有する。例えば、河川や海の富栄養化の原因となる有機化合物、窒素化合物、リン化合物の低減、悪臭の抑制、およびヌメリの原因物質の抑制といった極めて優れた効果を発揮できる。よって、当該細菌を用いる本発明は、環境を浄化できるものとして、産業上極めて有用である。 The bacterium of the present invention has an excellent treatment capacity for harmful compounds. For example, extremely excellent effects such as reduction of organic compounds, nitrogen compounds, and phosphorus compounds that cause eutrophication of rivers and seas, suppression of malodor, and suppression of causative substances of slime can be exhibited. Therefore, the present invention using the bacteria is extremely useful industrially as being capable of purifying the environment.
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