Chemistry: molecular biology and microbiology – Micro-organism – per se ; compositions thereof; proces of... – Bacteria or actinomycetales; media therefor
Reexamination Certificate
2001-03-23
2002-05-07
Prats, Francisco (Department: 1651)
Chemistry: molecular biology and microbiology
Micro-organism, per se ; compositions thereof; proces of...
Bacteria or actinomycetales; media therefor
C435S262000, C435S262500, C210S605000, C210S610000, C210S611000, C210S620000, C210S630000
Reexamination Certificate
active
06383797
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a novel bacterial consortium EBC1000 and a method for remedying biologically recalcitrant toxic chemicals contained in industrial wastewater, waste materials and soils using the bacterial consortium EBC1000. More particularly, the invention relates to a novel bacterial consortium EBC1000 capable of remarkably decomposing a chlorine compound of high concentration and waste acidic or alkaline recalcitrant toxic chemicals difficult to be degraded that are isolated from waste soil and water, the recalcitrant toxic chemicals having a chemical oxygen demand (COD
Mn
) of 20,000 to 100,000 ppm and comprising chemicals, such as isopropyl alcohol (IPA), methylene chloride (MC), dimethylamine (DMA), acrylonitrile (AN), hydrosulfite (SHS), butadiene (BD), sodium alkylaryl naphthalene sulfonate (Tamol-SN), tetra sodium ethylene diamine tetra acetate dihydrate (EDTA), ferrous sulfate heptahydrate (FES), tert-dodecyl mercaptan (TDDM), paramethane hydroperoxide (PMH), N-diethyl hydroxyl amine (DEHA), methanol (MeOH), NaOH and CH
3
CN.
BACKGROUND ART
Environmental contaminations, which are caused by recalcitrant toxic substances, often lead to (i) degradation of food stuffs for humans related to the food chain in the biosphere, (ii) prevalence of infection diseases due to deteriorated immunity, and (iii) spread of chronic diseases and energy exhaustion. Once the ecosphere is radically destroyed, it may take several hundred millions of years for the environment of the ecosphere to recover. degradation of food stuffs for human related to the food chain in the biosphere, prevalence of infectious diseases due to deteriorated immunity, spread of chronic diseases and energy exhaustion. Once the ecosphere is radically destroyed, it may take several hundred millions of years for recovery of the environment of ecosphere.
There has been an attempt to remedy recalcitrant toxic substances contained in the industrial wastewater or noxious waste materials dumped into the sea normally, using physical and chemical methods such as incineration, landfill, chemicals, electrolysis, membrane separation or the like. However, these conventional methods also involve economical and environmental problems. The most efficient and safe method that meets requirements in every aspect of environmentology, sanitation, ecology and economy is the biological treatment method. Many studies have been made on the biological methods, for example, using the natural ecosystem such as silt at an estuary. However, these methods could not be a solution of treating a great mass of industrial waste materials.
DISCLOSURE OF THE INVENTION
It is, therefore, an object of the present invention to provide bioremediation to remedy biologically recalcitrant toxic chemicals contaminating industrial wastewater, waste materials and soils, etc., using a trace of useful specific bacterial consortium that exists in the ecological system.
The present invention is directed to a novel bacterial consortium EBC1000 and a method for remedying biologically recalcitrant toxic chemicals and chlorine compounds contained in industrial wastewater, waste materials, soils, or the like using the bacterial consortium EBC1000. More particularly, the invention is directed to a novel bacterial consortium EBC1000 capable of remarkably degrading chlorine compounds of high concentration and waste acidic or waste alkaline recalcitrant toxic chemicals that are isolated from soil and wastewater having COD
Mn
of 20,000 to 100,000 ppm, such as contained in the medical industrial wastewater to be dumped into the sea, e.g., isopropyl alcohol, methylene chloride, NaOH, Na
2
SO
4
, dimethylamine or methanol, or as contained in the petroleum industrial wastewater to be dumped into the sea, e.g., sodium hydrosulfite, butadiene, sodium alkylaryl naphthalene sulfonate, acrylonitrile, tetra sodium ethylene diamine tetra acetate dihydrate, ferrous sulfate heptahydrate, tert-dodecyl mercaptan, paramethane hydroperoxide, N-diethyl hydroxy amine, and CH
3
CN.
The inventor isolated a novel bacterial consortium EBC1000 (KCTC 0652 BP) having a characteristic of rapidly degrading recalcitrant waste materials of high concentration dumped into the sea as well as chlorine compounds, from the soils and wastewater collected from the Ulsan industrial complex and the petroleum chemical industrial complex in Korea. The bacterial consortium EBC1000 consists of nine strains, each having the ability of degrading recalcitrant waste materials. It was found that the bacterial consortium EBC1000 degrades waste acidic or waste alkaline recalcitrant waste materials having the COD
Mn
of 20,000 to 100,000 ppm contained in the medical and petroleum industrial wastewaters to be dumped into the sea by 80% within 7 days, 90% within 10 days and at least 80 to 90% within 240 hours in an aerobic manner. Examples of the waste acidic or alkaline recalcitrant waste materials contained in the medical industrial wastewater of e.g., pH 3 or 14 include isopropyl alcohol (IPA, 10,000 to 20,000 ppm), methylene chloride (MC, 100 to 7,000 ppm), NaOH (changeable), Na
2
SO
4
(changeable), dimethylamine (DMA, 70 to 2,500 ppm), methanol (MeOH, 27,000 to 54,000 ppm), organic matters and antibiotic residues (40,000 to 100,000 ppm) and Cl
−
(4,000 to 33,000 ppm). Examples of the waste acidic or alkaline recalcitrant waste materials contained in the petroleum industrial wastewater of e.g., pH 1.2, 3.0 or 5.0 include sodium hydrosulfite (SHS, (NaSO
2
)
2
, 30 ppm), butadiene (BD,changeable), sodium alkylaryl naphthalene sulfonate (Tamol-SN, 4,000 to 20,000 ppm), acrylonitrile (AN,changeable), tetra sodium ethylene diamine tetra acetate dihydrate (EDTA, 20 to 200 ppm), ferrous sulfate heptahydrate (FES, 60 ppm), tert-dodecyl mercaptan (TDDM, 4,000 ppm), paramethane hydroperoxide (PMH, 400 ppm), N-diethyl hydroxyl amine (DEHA, 200 ppm), and CH
3
CN (changeable).
Accordingly, the present invention is to provide such a novel bacterial consortium EBC1000 capable of degrading recalcitrant toxic chemicals and chlorine compounds, and a method using the bacterial consortium EBC1000 in remedying biologically recalcitrant toxic chemicals and chlorine compounds contaminating industrial wastewater, waste materials and soils.
BEST MODE FOR CARRYING OUT THE INVENTION
Now, a description will be made to isolation, identification and activity of the novel bacterial consortium EBC1000.
1. Isolation of Novel Bacterial Consortium EBC1000
(1) Isolation of 40 Strains from Shaking Culture of the Samples
The soil (1 g) and wastewater (10 ml) from the Ulsan industrial complex were inoculated in a waste acidic and alkaline waste liquid medium (prepared by dilution of K
2
HPO
4
(0.65 g), KH
2
PO
4
(0.17 g), MgSO
4
(0.1 g), NaNO
3
(0.5 g) and 10% waste acidic and alkaline wastewater containing recalcitrant chemicals in medical and petroleum industrial waste liquid, in 1 liter of deionized water, pH 0 to 14) and then in a chlorine compound liquid medium (prepared by dissolving of K
2
HPO
4
(0.65 g), KH
2
PO
4
(0.17 g), MgSO
4
(0.1 g), NaNO
3
(0.5 g) and pentachlorophenol (PCP, 50 ppm) in 1 liter of deionized water, pH 7.2), followed by shaking culture at 20 to 30° C. for more than 5 days.
1 ml of the shaken culture was collected and inoculated in a waste acidic or waste alkaline solid medium (prepared by adding only 1.5% of agar to the waste acidic or alkaline liquid culture, pH 7.0) and then a chlorine compound solid medium (prepared by adding 20 mg of BTB and 1.5% of agar to the chlorine compound liquid medium), followed by incubation at 20 to 30° C. for 3 to 10 days.
The individual pure colonies isolated were further inoculated in the above mentioned cultures and then cultured with shaking, thus isolating 40 useful bacterial strains which have a different colony form formed respectively and the same colony after successive transfer culture.
(2) Isolation of 9 Component Strains After the Bacteria Isolated in Step (1) are cultured in the Medium of an Increased Concentration of the Recalcitrant Wa
Browdy and Neimark , P.L.L.C.
Prats Francisco
Srivastava Kailash C.
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