JP4836206B2 - Purification method of oil-containing soil and microorganism used therefor - Google Patents
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Description
本発明は、微生物を用いて油含有土壌を浄化する方法に関する。 The present invention relates to a method for purifying oil-containing soil using microorganisms.
油含有土壌の浄化方法には、土壌洗浄、熱脱着・熱分解、バイオレメディエーションなどの技術が存在する。これらのうち、バイオレメディエーションは、微生物により有害物質を二酸化炭素や水などの無害な物質に分解する技術であり、土着微生物を栄養塩や空気（酸素）などで活性化させ有害物質を分解させる方法（バイオスティミュレーション）と有害物質の分解に有効な微生物を選定・添加する方法（バイオオーグメンテーション）の２つに大別される。 Techniques for cleaning oil-containing soil include technologies such as soil cleaning, thermal desorption / pyrolysis, and bioremediation. Of these, bioremediation is a technology that decomposes harmful substances into harmless substances such as carbon dioxide and water by microorganisms, and is a method of activating indigenous microorganisms with nutrient salts and air (oxygen) to decompose harmful substances. (Biostimulation) and a method of selecting and adding microorganisms effective for decomposing harmful substances (bioaugmentation).
バイオオーグメンテーションに適用可能な微生物としては、例えば、特許文献１や特許文献２などに種々記載されている。 Various microorganisms applicable to bioaugmentation are described in, for example, Patent Literature 1 and Patent Literature 2.
バイオオーグメンテーションの実用化にあたっては、使用する微生物が周辺環境への影響がなく安全性の高いことが要求される。 In putting bioaugmentation into practical use, it is required that the microorganisms used have high safety without affecting the surrounding environment.
本発明は、上記事情に鑑みてなされたものであり、その課題は、外来の微生物を用いて油含有土壌を浄化する新規の方法およびそれに用いる微生物を提供することにある。 The present invention has been made in view of the above circumstances, and an object thereof is to provide a novel method for purifying oil-containing soil using a foreign microorganism and a microorganism used therefor.
本発明者は、上記課題を解決するため鋭意検討した結果、Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）を単離し、これらがいずれも鉱油類を含む油含有土壌に対して分解能力に優れることを見出し、本発明を完成した。 As a result of intensive studies to solve the above problems, the present inventor has found that Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus sp. No. 10 A strain (FERM AP-21568) was isolated, and all of them were found to have excellent decomposability against oil-containing soils containing mineral oils, thereby completing the present invention.
すなわち、本発明の要旨は以下のとおりである。
〔１〕 Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）のうち、少なくとも１種の微生物を油含有土壌に添加することを特徴とする油含有土壌の浄化方法、
〔２〕 Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）を油含有土壌に添加する、前記〔１〕記載の油含有土壌の浄化方法、
〔３〕 油含有土壌が、原油、重油、軽油、灯油、ガソリン、潤滑油、切削油、作動油、圧延油、絶縁油、エンジンオイル、ギアオイル、グリースから選ばれる鉱油類を含有するものである、前記〔２〕記載の油含有土壌の浄化方法、
〔４〕 鉱油類が、炭素数Ｃ１２〜Ｃ２８の油分を含有するものである、前記〔３〕記載の油含有土壌の浄化方法、
〔５〕 前記〔１〕〜〔４〕のいずれか記載の方法に用いられる、Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）のうち、少なくとも１種の微生物。
That is, the gist of the present invention is as follows.
[1] At least one of Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus sp. No. 10 strain (FERM AP-21568) A method for purifying oil-containing soil, characterized by adding the microorganisms to oil-containing soil,
[2] Add Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus sp. No. 10 strain (FERM AP-21568) to oil-containing soil The method for purifying oil-containing soil according to [1],
[3] Oil-containing soil contains mineral oil selected from crude oil, heavy oil, light oil, kerosene, gasoline, lubricating oil, cutting oil, hydraulic oil, rolling oil, insulating oil, engine oil, gear oil, and grease. The method for purifying oil-containing soil according to [2],
[4] The method for purifying oil-containing soil according to [3] above, wherein the mineral oil contains an oil component having a carbon number of C12 to C28.
[5] Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus used in the method according to any one of [1] to [4] At least one microorganism of sp. No. 10 strain (FERM AP-21568).
本発明によれば、鉱油類を含む油含有土壌を効率的に浄化する方法およびそれに用いる微生物が提供される。 According to the present invention, a method for efficiently purifying oil-containing soil containing mineral oils and a microorganism used therefor are provided.
本発明に係る油含有土壌の浄化方法は、上述したように、Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）のうち、少なくとも１種の微生物を油含有土壌に添加することを特徴とする。 As described above, the method for purifying oil-containing soil according to the present invention includes Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus sp. Among 10 strains (FERM AP-21568), at least one microorganism is added to the oil-containing soil.
本発明において「油含有土壌」とは、原油、重油、軽油、灯油、ガソリン、潤滑油、切削油、作動油、圧延油、絶縁油、エンジンオイル、ギアオイル、グリースなどの石油または石油を原料とする有機物（以下、これらをまとめて「鉱油類」という）を含む土壌をいい、例えば、石油により汚染された海岸の砂および土壌、石油により汚染された工場やガソリンスタンドの跡地の土壌、石油により汚染された廃棄物最終処分場の土壌、及び油田等の天然の状態で石油を含有する土壌を含むものである。 In the present invention, “oil-containing soil” refers to crude oil, heavy oil, light oil, kerosene, gasoline, lubricating oil, cutting oil, hydraulic oil, rolling oil, insulating oil, engine oil, gear oil, grease, etc. This refers to soil containing organic matter (hereinafter collectively referred to as “mineral oils”). For example, coastal sand and soil contaminated with oil, soil of factory and gas station sites contaminated with oil, and oil This includes soil in the final disposal site of contaminated waste and soil containing oil in a natural state such as oil fields.
本発明においては、上記「油含有土壌」の中でも、炭素数Ｃ６〜Ｃ４４の油分（以下、「Ｃ６〜Ｃ４４成分」と略記する。他の炭素数の油分を表記するときも同様とする）を含有する油含有土壌、特にＣ１２〜Ｃ２８を主成分として含有する油含有土壌をより効率的に浄化することができる。なお、Ｃ１２〜Ｃ２８成分は軽油、重油等の主成分である。また、本発明において「浄化」とは、残留油分、油臭及び油膜の低減又は除去を意味する。 In the present invention, among the above-mentioned “oil-containing soil”, an oil component having a carbon number of C6 to C44 (hereinafter abbreviated as “C6 to C44 component”. The same applies when an oil component having another carbon number is described). The oil-containing soil to contain, especially the oil-containing soil which contains C12-C28 as a main component can be purified more efficiently. In addition, C12-C28 components are main components, such as light oil and heavy oil. In the present invention, “purification” means reduction or removal of residual oil, oily odor and oil film.
本発明に適用可能な微生物は、Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）およびRhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）のうち、少なくとも１種である。上記３種の微生物は、油田や油汚染土壌を分離源とし、重油を唯一の炭素源とした集積培養を経た後、寒天培地よりコロニーを単離することにより得られる。これら３種の単離菌の菌学的性質は表１及び表２に記載の通りである。 The microorganisms applicable to the present invention include Novosphingobium sp. No. 2 strain (FERM AP-21666), Pseudomonas sp. No. 5 strain (FERM AP-21567) and Rhodococcus sp. No. 10 strain (FERM AP-21568). Among them, at least one. The above three types of microorganisms can be obtained by isolating colonies from an agar medium after an enrichment culture using an oil field or oil-contaminated soil as a separation source and heavy oil as the sole carbon source. The bacteriological properties of these three types of isolated bacteria are as shown in Tables 1 and 2.
図１〜図３は、上記３種の単離菌とその近縁種の１６Ｓ ｒＲＮＡの遺伝子塩基配列に基づいて作成した系統樹である。Novosphingobium sp. No.2株（ＦＥＲＭ ＡＰ−２１５６６）（図１において、「No.2」と表記）はNovosphingobium属細菌との相同性が高く、最近縁種であるNovosphingobium capsulatumとの相同性が９８％であることから、Novosphingobium sp. No.2株はNovosphingobium capsulatumと同種または異種ながらも非常に近い性質をもつ菌であると考えられる。 1 to 3 are phylogenetic trees prepared based on the gene base sequences of 16S rRNA of the above three types of isolated bacteria and related species. Novosphingobium sp. No.2 strain (FERM AP-21565) (referred to as “No.2” in FIG. 1) has high homology with Novosphingobium genus bacteria and has a homology with Novosphingobium capsulatum, which is a recent relative, 98 Therefore, Novosphingobium sp. No.2 strain is considered to be a fungus having the same or different characteristics as Novosphingobium capsulatum but very close.
また、Pseudomonas sp. No.5株（ＦＥＲＭ ＡＰ−２１５６７）（図２において、「No.5」と表記）はPseudomonas属細菌との相同性が高く、最近縁種であるPseudomonas citronellolisとの相同性が９９％であることから、Pseudomonas sp. No.5株はPseudomonas citronellolisと同種または異種ながらも非常に近い性質をもつ菌であると考えられる。 In addition, Pseudomonas sp. No. 5 strain (FERM AP-21567) (indicated as “No. 5” in FIG. 2) has high homology with bacteria belonging to the genus Pseudomonas, and homology with Pseudomonas citronellolis, which is a recent relative. Is 99%, Pseudomonas sp. No.5 strain is considered to be a fungus having the same or different characteristics as Pseudomonas citronellolis but very close.
さらに、Rhodococcus sp. No.10株（ＦＥＲＭ ＡＰ−２１５６８）（図３において、「No.10」と表記）はRhodococcus属細菌との相同性が高く、最近縁種であるRhodococcus erythropolisとの相同性が９９％であることから、Rhodococcus sp. No.10株はRhodococcus erythropolisと同種または異種ながらも非常に近い性質をもつ菌であると考えられる。 Furthermore, Rhodococcus sp. No. 10 strain (FERM AP-21568) (indicated as “No. 10” in FIG. 3) has high homology with bacteria belonging to the genus Rhodococcus, and homology with Rhodococcus erythropolis which is a recent relative. Is 99%, Rhodococcus sp. No. 10 strain is considered to be a bacterium having the same or different characteristics as Rhodococcus erythropolis but having very close characteristics.
上記３種の単離菌をそれぞれ単独でＣ６〜Ｃ４４成分を含有する油含有土壌に添加すると、該土壌中の油分が効率的に分解される。また、上記３種の単離菌全てをＣ６〜Ｃ４４成分を含有する油含有土壌に添加すると、それぞれを単独で添加したときと比べて、上記土壌中の油分がさらに効率的に分解される。すなわち、上記３種の単離菌を全て油含有土壌に添加した場合、油分の分解効率について相乗効果を発揮する。 When each of the three types of isolated bacteria is added to oil-containing soil containing C6-C44 components alone, the oil in the soil is efficiently decomposed. Moreover, when all the three types of isolated bacteria are added to the oil-containing soil containing the C6 to C44 components, the oil content in the soil is more efficiently decomposed than when each of them is added alone. That is, when all the three types of isolated bacteria are added to the oil-containing soil, a synergistic effect is exhibited with respect to the decomposition efficiency of the oil.
本発明において、上記単離菌を油含有土壌に「添加する」とは、上記単離菌を油含有土壌と接触させることをいい、上記単離菌を油含有土壌に添加する手段の他、例えば、投入する、供給する、散布する、土壌中に注入する等の方法も含む概念である。 In the present invention, "adding" the isolated bacteria to the oil-containing soil means contacting the isolated bacteria with the oil-containing soil, in addition to means for adding the isolated bacteria to the oil-containing soil, For example, it is a concept including methods such as charging, supplying, spraying, and pouring into soil.
油含有土壌の浄化を効率良く進めるためには、上記単離菌の好適な増殖条件となるように、上記単離菌を油含有土壌に添加すること以外に、例えば、該土壌を撹拌したり、栄養塩類を添加したり、該土壌に水分を添加したり、土壌に注入する場合には併せて空気も注入するなどして、該土壌中の酸素供給濃度、温度、水分、塩類濃度などを適宜調整することが好ましい。 In order to efficiently purify the oil-containing soil, in addition to adding the isolated bacteria to the oil-containing soil, for example, stirring the soil so as to achieve suitable growth conditions for the isolated bacteria. When adding nutrients, adding moisture to the soil, or injecting air into the soil, injecting air together, the oxygen supply concentration, temperature, moisture, salt concentration, etc. in the soil can be adjusted. It is preferable to adjust appropriately.
上記単離菌は、油含有土壌中の油分のうち、特にＣ１２〜Ｃ２８成分をより効率的に分解することができる。したがって、軽油や重油等を主成分とする油含有土壌に対して上記単離菌を添加すれば、本発明の効果が最も発揮される。 The said isolate can decompose | disassemble especially C12-C28 component more efficiently among the oil components in oil-containing soil. Therefore, if the said isolation microbe is added with respect to the oil-containing soil which has light oil, heavy oil, etc. as a main component, the effect of this invention is exhibited most.
以下、実施例により本発明をさらに詳しく説明するが、本発明はこれらのみに限定されるものではない。なお、以下において、「％」は、特にことわりのない限り、重量％を示す。 EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited only to these. In the following, “%” indicates wt% unless otherwise specified.
１．微生物の単離
１−１． 単離菌１（Novosphingobium sp. No.2株）の単離
単離菌１は、新潟県柏崎市の油田を分離源とし、重油を唯一の炭素源とした集積培養を経た後、寒天培地よりコロニーを単離することにより得られた。単離菌１の１６Ｓ ｒＲＮＡ遺伝子塩基配列を比較した結果（図１参照）、単離菌１（図１において、「No.2」と表記）は、Novosphingobium属細菌との相同性が高く、最近縁種はNovosphingobium capsulatumであった（相同性９８％）。以上の結果より、単離菌１はNovosphingobium属に属する細菌であることを確認した。単離菌１はNovosphingobium sp. No.2株と命名され、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にＦＥＲＭ ＡＰ−２１５６６として寄託されている。
1. 1. Isolation of microorganisms 1-1. Isolation of isolated bacterium 1 (Novosphingobium sp. No.2 strain) Isolated bacterium 1 was obtained from an agar medium after an enrichment culture using an oil field in Amagasaki City, Niigata Prefecture and heavy oil as the sole carbon source. Obtained by isolating colonies. As a result of comparison of 16S rRNA gene base sequences of isolate 1 (see FIG. 1), isolate 1 (indicated as “No. 2” in FIG. 1) has high homology with Novosphingobium bacteria, The relative species was Novosphingobium capsulatum (98% homology). From the above results, it was confirmed that the isolated bacterium 1 is a bacterium belonging to the genus Novosphingobium. The isolated bacterium 1 is named Novosphingobium sp. No. 2 strain and has been deposited as FERM AP-21666 at the National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center.
１−２． 単離菌２（Pseudomonas sp. No.5株）の単離
単離菌２は、新潟県北蒲原郡の油田を分離源とし、重油を唯一の炭素源とした集積培養を経た後、寒天培地よりコロニーを単離することにより得られた。単離菌２の１６Ｓ ｒＲＮＡ遺伝子塩基配列を比較した結果（図２参照）、単離菌２（図２において「No.5」と表記）は、Pseudomonas 属細菌との相同性が高く、最近縁種はPseudomonas citronellolisであった（相同性９９％）。以上の結果より、単離菌２はPseudomonas属に属する細菌であることを確認した。単離菌２はPseudomonas sp. No.5株と命名され、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にＦＥＲＭ ＡＰ−２１５６７として寄託されている。
1-2. Isolation of isolated bacterium 2 (Pseudomonas sp. No.5 strain) Isolated bacterium 2 is obtained from an agar medium after an enrichment culture using the oil field of Kitaharabara-gun, Niigata Prefecture and heavy oil as the sole carbon source. Obtained by isolating colonies. As a result of comparison of 16S rRNA gene base sequences of isolated bacteria 2 (see FIG. 2), isolated bacteria 2 (indicated as “No. 5” in FIG. 2) has high homology with Pseudomonas bacteria, The species was Pseudomonas citronellolis (99% homology). From the above results, it was confirmed that the isolated bacterium 2 is a bacterium belonging to the genus Pseudomonas. The isolated bacterium 2 is named Pseudomonas sp. No.5 strain and is deposited as FERM AP-21567 at the National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center.
１−３． 単離菌３（Rhodococcus sp. No.10株）の単離
単離菌３は、神奈川県川崎市の土壌を分離源とし、重油を唯一の炭素源とした集積培養を経た後、寒天培地よりコロニーを単離することにより得られた。単離菌３の１６Ｓ ｒＲＮＡ遺伝子塩基配列を比較した結果（図３参照）、単離菌３（図３において「No.10」と表記）は、Rhodococcus 属細菌との相同性が高く、最近縁種はRhodococcus erythropolisであった（相同性９９％）。以上の結果より、単離菌３はRhodococcus属に属する細菌であることを確認した。単離菌３はRhodococcus sp. No.10株と命名され、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にＦＥＲＭ ＡＰ−２１５６８として寄託されている。
1-3. Isolation of isolated bacterium 3 (Rhodococcus sp. No.10 strain) Isolated bacterium 3 is obtained from an agar medium after being subjected to enrichment culture using soil in Kawasaki City, Kanagawa Prefecture as the separation source and heavy oil as the sole carbon source. Obtained by isolating colonies. As a result of comparing the base sequence of 16S rRNA gene of isolated bacterium 3 (see FIG. 3), isolated bacterium 3 (indicated as “No. 10” in FIG. 3) has high homology with Rhodococcus genus bacteria. The species was Rhodococcus erythropolis (99% homology). From the above results, it was confirmed that the isolated bacterium 3 is a bacterium belonging to the genus Rhodococcus. The isolated bacterium 3 is named Rhodococcus sp. No.10 strain, and is deposited as FERM AP-21568 at the National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center.
２．単離菌１〜３の菌学的性質
AP120NE及びATB BET（ともに bioMerieux社）を用いて単離菌１〜３の保持酵素、同化性及び抗生物質感受性を調べた。結果を表１及び表２に示す。
2. Mycological properties of isolated bacteria 1-3
AP120NE and ATB BET (both bioMerieux) were used to examine the retained enzymes, assimilation, and antibiotic susceptibility of isolates 1-3. The results are shown in Tables 1 and 2.
３．油分分解試験（実施例１〜４、比較例１）
油分濃度（Ｃ６〜Ｃ４４の全成分濃度）０．４３〜０．４９％の油含有土壌に上記で単離したNovosphingobium sp. No.2株（単離菌１）、Pseudomonas sp. No.5株（単離菌２）、Rhodococcus sp. No.10株（単離菌３）のうち、１種または３種を投入し、水分と栄養塩を供給し、上記土壌中の油分濃度の経時変化を調べた。油分濃度は、赤外分光分析法（ＩＲ法）により定量した（実施例１〜４）。
3. Oil decomposition test (Examples 1-4, Comparative Example 1)
Novosphingobium sp. No.2 strain (isolated strain 1), Pseudomonas sp. No.5 strain isolated above in oil-containing soil with oil content (concentration of all components of C6-C44) of 0.43-0.49% (Isolated Bacteria 2), Rhodococcus sp. No. 10 (Isolated Bacteria 3), 1 or 3 species are added, water and nutrients are supplied, and the oil concentration in the soil is changed over time. Examined. The oil concentration was quantified by infrared spectroscopy (IR method) (Examples 1 to 4).
下記に具体的試験方法、菌の投入量、栄養塩の種類、添加量を示す。
（１）試験方法：某地油汚染土壌０．７ｋｇ（油分濃度０．４３〜０．４９％）をアルミバット（縦４０ｃｍ×横３０ｃｍ×高さ５ｃｍ）中に四角錘台状に盛り上げ、含水比が常に１５％に保たれるよう、適宜加水した。
（２）試験温度：室内、２０℃
（３）菌株の投入量：単離菌１〜３のうち、１種のみを投入する場合は、各菌株をそれぞれ２×１０６cell／ｇ（乾土）で投入した。上記３種の単離菌を混合投入する場合は、各菌株をそれぞれ０．６７×１０６cell／ｇ（乾土）（合計２×１０６cell／ｇ（乾土））ずつ投入した。
（４）栄養塩類の種類：窒素（硫安）、リン（過リン酸石灰）
（５）栄養塩類の添加量：窒素２５０ｐｐｍ、リン５０ｐｐｍ
（６）油分濃度の測定日：試験開始時、５、８、１４日後
The specific test method, the amount of bacteria input, the type of nutrient, and the amount added are shown below.
(1) Test method: 0.7 kg (oil concentration: 0.43 to 0.49%) of dredged oil-contaminated soil was raised into a square frustum in an aluminum bat (
(2) Test temperature: Indoor, 20 ° C
(3) Amount of strain input: When only one of the isolated bacteria 1 to 3 was input, each strain was input at 2 × 10 6 cells / g (dry soil). When the above three kinds of isolated bacteria were mixed and added, each strain was added at 0.67 × 10 6 cell / g (dry soil) (total 2 × 10 6 cell / g (dry soil)).
(4) Types of nutrients: nitrogen (ammonium sulfate), phosphorus (superphosphate lime)
(5) Nutrient addition amount: nitrogen 250ppm, phosphorus 50ppm
(6) Date of oil concentration measurement: 5, 8, 14 days after the start of the test
比較例１として、上記単離菌を投入しないこと以外は、上記と同様の方法で上記土壌中の油分濃度の経時変化を調べた。結果を表３および図４に示す。 As Comparative Example 1, the change over time in the oil concentration in the soil was examined by the same method as above except that the isolated bacterium was not added. The results are shown in Table 3 and FIG.
表３及び図４の結果から、単一の単離菌を投入した実施例１〜３のいずれも、比較例１に比べて油分濃度の低減率が大きく、油分の分解が促進されることが分かった。このことから、上記土壌中に存在する土着菌のみの分解に比べて、単離菌１〜３を加えた方が油分の分解能力に優れているといえる。
また、単離菌１〜３を全て投入した実施例４は、実施例１〜３に比べて油分濃度の低減率が大きく、油分の分解がさらに促進されることが分かった。具体的には、実施例４では、油分の低減率が、８日後で約３０％であり、１４日後で約４３％であった。
From the results of Table 3 and FIG. 4, all of Examples 1 to 3 into which a single isolated bacterium was introduced had a large reduction rate of the oil concentration compared to Comparative Example 1 and promoted the decomposition of the oil. I understood. From this, it can be said that the addition of the isolated bacteria 1 to 3 is superior in the ability to decompose oil compared to the decomposition of only the indigenous bacteria present in the soil.
Moreover, it turned out that Example 4 which injected all isolated bacteria 1-3 has a large reduction rate of an oil concentration compared with Examples 1-3, and decomposition | disassembly of an oil content is further accelerated | stimulated. Specifically, in Example 4, the oil reduction rate was about 30% after 8 days and about 43% after 14 days.
表４は、本試験で使用した油含有土壌について、試験６ヶ月前に測定した全石油系炭化水素（ＴＰＨ）分析による組成分析結果である。表４によれば、本試験の６ヶ月前の油分濃度（表４の「全炭化水素量」を参照）は０．６９％であり、表３の試験開始時の油分濃度（表３の「初期値」を参照）が０．４３〜０．４９％であるから、本試験を行うまでの６ヶ月の間に油分濃度が約０．２％減少している。これは、まず炭素数が少ないＣ６〜Ｃ１２成分が分解され、続いてＣ１２〜Ｃ２８成分が一部分解されたことによるものと推定される。したがって、本試験に使用した油含有土壌中に存在する油分のうち、主成分はＣ１２〜Ｃ２８成分であり、油分全体の約９５％であると考えられる。 Table 4 shows the composition analysis result by total petroleum hydrocarbon (TPH) analysis measured 6 months before the test for the oil-containing soil used in this test. According to Table 4, the oil concentration 6 months before this test (see “Total amount of hydrocarbons” in Table 4) is 0.69%, and the oil concentration at the start of the test in Table 3 (“ Since the initial value is 0.43 to 0.49%, the oil concentration is reduced by about 0.2% during the six months until the test is conducted. This is presumably because the C6-C12 component having a small number of carbons was first decomposed and then the C12-C28 component was partially decomposed. Therefore, among the oils present in the oil-containing soil used in this test, the main component is the C12 to C28 component, which is considered to be about 95% of the total oil.
そうすると、上記の試験結果から、本試験で使用した単離菌１〜３は、いずれも油分のうち、特にＣ１２〜Ｃ２８成分の分解能力に優れた微生物（油分解菌）であるということがいえる。 Then, from the above test results, it can be said that the isolated bacteria 1 to 3 used in this test are microorganisms (oil-decomposing bacteria) that are particularly excellent in the ability to decompose C12 to C28 components in the oil. .
本発明は、鉱油類を含有する油含有土壌、特にＣ１２〜Ｃ２８成分を含有する油含有土壌をより効率的に浄化する方法およびそれに用いる微生物として広く利用することができる。 INDUSTRIAL APPLICABILITY The present invention can be widely used as a method for efficiently purifying oil-containing soil containing mineral oils, particularly oil-containing soil containing C12 to C28 components, and microorganisms used therefor.
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