JP2001238694A - Method for microbiologically identifying chemical substance - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for microbiologically identifying a sample chemical substance, because information for judging the property of toxicity or the kind of chemical substances causing the toxicity is not entirely afforded, though the bioassay utilized at present mainly measures only growth inhibition of unicell and the presence of the toxicity due to the chemical substances can be evaluated. SOLUTION: This method for microbiologically identifying chemical substances features comprising a microorganism growth process for growing previously decided several kinds of marker microorganisms in the presence of the sample chemical substance under various concentrations of the sample chemical substance, a measurement process for measuring the relationship between the concentration of the sample chemical substance and the concentration of the marker microorganisms on the microorganism growth process and a comparison process for comparing the relationship between the concentration of the marker microorganisms and the concentration of the sample chemical substance obtained by the measurement process with the data showing the relationship between the concentration of a marker microorganism previously put away into a data bank and the concentration of a marker chemical substance.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for microbiologically identifying a chemical substance.

[0002]

2. Description of the Related Art At present, a chemical substance database Chemic
About 17 million chemical substances are registered in al abstract. Among them, it is estimated that more than 10,000 kinds of synthetic chemicals are accumulated in the environment, and the number is increasing year by year. Some synthetic chemicals have a negative effect on the ecology and the human body after being transformed directly or in the environment. Therefore, it is necessary to quickly evaluate the effects of each chemical substance on ecology and the human body. Also,
Under the situation where the environmental pollution problem has become a social problem that raises public unrest, attempts have been made to identify and quantify chemical substances present in the environment with various advanced measurement systems. However, it is said that at present, only about 10% of chemical substances can be identified using any advanced technology. Therefore, it is more practical to establish an index capable of expressing the toxicity of a chemical substance existing in the environment than to identify the unknown chemical substance itself existing in the environment. Due to the above-mentioned necessity, multi-institutions are now trying to index toxicity using a simple toxicity evaluation test (bioassay). The bioassay is a method for evaluating "toxicity" by measuring "change in biological response due to a chemical substance" using animal and plant cells or microorganisms without using an individual organism. However, currently used bioassays mainly measure single cell growth inhibition. In this method, the presence or absence of toxicity due to a chemical substance can be evaluated, but no information for determining the nature of the toxicity or the type of toxicity caused by the chemical substance can be obtained at all.

[0003]

An object of the present invention is to provide a method for microbiologically identifying a sample chemical substance.

[0004]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems, and as a result, have completed the present invention. That is, according to the present invention, a microorganism growth step of growing a plurality of predetermined indicator microorganisms by changing the concentration of the sample chemical substance in the presence of the sample chemical substance,
A measurement step of measuring the relationship between the sample chemical substance concentration and the indicator microorganism concentration in the microorganism growth step, and the relationship between the indicator microorganism concentration and the sample chemical substance concentration obtained in the measurement step is stored in a data bank in advance. And a collation step of collating with data indicating a relationship between the concentration of the indicator microorganism and the concentration of the indicator chemical substance.

[0005]

DETAILED DESCRIPTION OF THE INVENTION The method of the present invention uses a data bank for microbiologically identifying the sample chemical. This data bank stores data indicating the relationship between the indicator chemical substance concentration and the indicator microorganism concentration measured in advance. It is preferable that the number of data (hereinafter simply referred to as microorganism / chemical substance concentration data) indicating the relationship between the concentration of the index chemical substance and the concentration of the index microorganism stored in this data bank be as large as possible. Is newly obtained, it is preferable to quickly store the data in the data bank. In the case of the present invention, it is preferable to generate microorganism / chemical substance concentration data using a plurality of predetermined indicator microorganisms and a plurality of predetermined indicator chemical substances, and store the data in a data bank. .

[0006] As the indicator chemical substance, a substance showing an inhibitory property against microorganisms is used. As such a substance, a substance exhibiting the following growth inhibitory properties against Escherichia coli was selected. The minimum inhibitory concentration against E. coli is shown in parentheses after the compounds described below.

[0007] No. 1 Methylmercury chloride
(0.91 μM) No. 2 Nickel chloride (1111 μM) No. 32 2-Aminoanthracene (370 μM) 4 Tributyltin chloride (0.1 μM) No. 5 Malathion (222 μM) No. 5 6 Phenol (17 mM) No. 7 Lindane (6666 μM) No. 7 8 Pentachlorophenol (74 μM) No. 9 Trp-P-2 (Acetate) (37 μM) No. 10 4-Nitroquinoline-N-oxide (567 μM) 11 Benzo (a) pyrene (111 μM) 12 Paraquat (333 μM) 13 Maneb (37 μM) No. 14 Cadmium chloride (333 μM) No. 15 Bis phenol-A (123 μM) No. 16 Di-2-ethylhexyl phthalate (92.6μ
M) No. 17 2,5-Dichlorophenol (13.7 μM) No. 18 2,4-Dichlorophenoxy acetic acid (24
7 μM). 19 Formaldehyde (555 μM) No. 20 p-Nonylphenol (666 μM) No. 21 Sodium Arsenite (111 μM) No. 22 Thiuram (12.3 μM) 23 2,4,5-Trichlorophenol (13.7 μM) No. 24 potassium dichlomate (33 μM) 25 Triphenyltin chloride (3.3 μM)

[0008] Although 25 kinds of indicator chemicals are shown in the above, it is clear that it is preferable to select as many chemicals as possible in order to perform more accurate microbiological identification of chemicals. . Therefore, it is natural that the indicator chemical is not limited to the above.

As the indicator microorganisms, it is preferable to select a large number of different types of indicator microorganisms. Specific examples of the indicator microorganisms are as follows. (1) Acetobacter pasteurianus IFO 3188 This microorganism is methylmercury chlori
It is a microorganism that is sensitive to de). The growth of this microorganism is almost completely inhibited in the presence of about 1 μM of methylmercury chloride, and even at a concentration of about 0.3 μM, obvious growth inhibition occurs. Sensitivity to other chemicals is shown in Table 1. (2) Acetobacter pasteurianus IFO 3129 This microorganism is a microorganism that is sensitive to nickel chloride. This microorganism showed only about 60% growth in the presence of about 0.17 μM nickel chloride. Sensitivity to other chemicals is shown in Table 2. (3) Glysomyces rutagersensis IFO 14488 This microorganism is a microorganism showing sensitivity to 2-aminoanthracene. This microorganism is sensitive even at a concentration of about 4.6 μM of 2-aminoanthracene. Sensitivity to other chemicals is shown in Table 3. (4) Aureobacterium esteraromatium IFO 3752 This microorganism is a microorganism that is sensitive to tributyltin chloride. This microorganism is sensitive to tributyltin chloride up to concentrations as low as 0.003 μM. The sensitivity to other chemicals is shown in Table 4. (5) Comamonas Testosteroni IAM 1048 This microorganism is a microorganism that is susceptible to malathion. This microorganism caused obvious growth inhibition in the presence of about 0.3 μM malathion. Sensitivity to other chemicals is shown in Table 5. (6) Pseudomonas synxantha IFO 3913 This microorganism is a microorganism showing sensitivity to phenol. This microorganism showed obvious growth inhibition at a phenol concentration of about 23 μM. Sensitivity to other chemicals is shown in Table 6. (7) Staphylococcus aureus IFO 3060 This microorganism is a microorganism showing sensitivity to Lindane. This microorganism showed growth inhibition even at about Lindan 0.34 μM. Sensitivity to other chemicals is shown in Table 7. The seven types of microorganisms described above have different sensitivities to chemical substances, and show strong sensitivities to different chemical substances.

The growth (cultivation) of the indicator microorganisms was carried out using an L medium (1% yeast extract, 0.5% polypeptone). The growth was performed at 25 ° C. on a 96-well microplate, and the growth (microorganism concentration) was observed at an absorbance of 650 nm. In this case, a predetermined chemical substance was added to the medium at a predetermined concentration. Late Tables 1 to 7 show the sensitivity (microorganism / chemical substance concentration data) of the indicator microorganism to the indicator chemical. In Tables 1 to 7 below, the values of the absorbance at 650 nm shown in the column of growth correspond to the values of the concentration (μM) of the chemical substance shown in the upper column. For example, in Table 1, (1) Acetobacter pasteurian
us Absorbance 102 and 0.1 in the column of growth in IFO3188
16 is the concentration of methylmercury chloride (μ
M) Corresponds to 0.000 and 0.034.

[0011]

(Table 1) (1) Chemical sensitivity of Acetobacter pasteurianus IFO3188 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.102 0.116 0.108 0.085
0.024 0.021 0.0240.025 0.024 0.024 0.018 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.109 0.113 0.112 0.119
0.114 0.114 0.1140.117 0.100 0.012 0.013 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.101 0.109 0.116 0.110
0.107 0.106 0.0900.089 0.065 -0.048 -0.091 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.104 0.049 0.048 0.049
0.046 0.040 0.0180.016 0.011 0.015 0.024Malathion
Concentration (μM) 0.000 0.034 0.102 0.305 0.914 2.741 8.
222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.111 0.111 0.127 0.120
0.126 0.120 0.1230.120 0.108 0.081 -0.003 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.107 0.112 0.124 0.108
0.107 0.099 0.0800.075 0.038 0.027 0.018 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.108 0.111 0.119 0.121 0.1
18 0.085 0.0700.094 0.100 0.094 -0.063 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.104 0.113 0.123 0.111
0.086 0.037 0.0180.018 0.018 0.017 0.014 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.098 0.120 0.121 0.110
0.111 0.102 0.0740.061 0.039 0.015 0.074 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.121 0.114 0.121 0.112
0.120 0.109 0.1040.098 0.082 0.018 0.014 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.093 0.090 0.121 0.120
0.120 0.115 0.1110.095 0.081 0.060 0.051 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.110 0.107 0.114 0.107
0.103 0.093 0.0750.059 0.024 0.017 0.015 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.111 0.116 0.127 0.123
0.125 0.120 0.1140.085 0.014 0.009 0.018 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.106 0.098 0.115 0.115
0.111 0.102 0.0900.055 0.020 0.017 0.016 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.090 0.095 0.121 0.125
0.119 0.112 0.0950.063 0.021 0.020 0.070 Di-2-ethylhexyl phthalate concentration (μM) 0.000 0.042
0.127 0.381 1.143 3.42810.284 30.852 92.556 277.66
7 833.000 Growth (Absorbance at 650nm) 0.091 0.087 0.118 0.114
0.118 0.116 0.1100.094 0.066 0.047 0.0522,5-Dichl
orophenol concentration (μM) 0.000 0.169 0.508 1.524 4.572
13.716 41.148 123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.098 0.090 0.121 0.122
0.126 0.127 0.1060.070 0.021 0.017 0.017 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.099 0.098 0.118 0.126
0.119 0.121 0.1150.107 0.081 0.018 0.021 Formaldeh
yde concentration (μM) 0.000 0.085 0.254 0.762 2.285 6.856
20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.109 0.098 0.116 0.116
0.115 0.114 0.1120.086 0.048 0.019 0.015 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.108 0.086 0.126 0.126
0.112 0.020 0.0140.011 0.012 0.024 0.099 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.111 0.113 0.121 0.122
0.125 0.121 0.1230.112 0.047 0.026 0.009Thiuram
Concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.370 4.1
11 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.112 0.107 0.113 0.106
0.105 0.080 0.0500.027 0.018 0.013 0.013 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.118 0.113 0.122 0.109
0.065 0.019 0.0180.018 0.016 0.020 0.027 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.115 0.107 0.114 0.113
0.111 0.110 0.1160.115 0.112 0.104 0.079 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.107 0.112 0.110 0.106
0.105 0.106 0.1000.108 0.104 0.091 0.083

[0012]

[Table 2] (2) Chemical sensitivity of Acetobacter pasteurianus IFO3129 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.260 0.313 0.292 0.269
0.165 0.036 0.0470.047 0.041 0.034 0.032 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.556 0.345 0.265 0.264
0.261 0.265 0.5840.272 0.480 0.114 0.026 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.272 0.296 0.284
0.269 0.267 0.264 0.2370.198 0.166 0.031 -0.040 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.278 0.047 0.047 0.047
0.046 0.046 0.0390.025 0.013 0.016 0.030 Malathion concentration (μM) 0.000 0.034 0.102 0.305 0.914
2.741 8.222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.265 0.252 0.255 0.253
0.243 0.230 0.2310.223 0.214 0.168 0.083 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.323 0.300 0.323 0.305
0.289 0.283 0.2670.224 0.178 0.071 0.032 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.234 0.234 0.235 0.235
0.221 0.168 0.1370.164 0.158 0.132 -0.114 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.271 0.228 0.213 0.199
0.155 0.073 0.0290.021 0.023 0.021 0.021 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.648 0.580 0.288 0.285
0.274 0.140 0.0700.036 0.023 0.023 0.068 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.296 0.299 0.298 0.303
0.303 0.289 0.2810.240 0.154 0.021 0.024 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.077 0.079 0.078 0.079
0.076 0.074 0.0710.054 0.047 0.047 0.048 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.349 0.311 0.314 0.276
0.236 0.193 0.1560.123 0.049 0.032 0.024 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.341 0.323 0.371 0.332
0.671 0.601 0.5970.250 0.642 0.059 0.027 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.252 0.247 0.245 0.227
0.210 0.192 0.1600.120 0.021 0.017 0.024 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.246 0.257 0.242 0.250
0.242 0.225 0.2010.090 0.030 0.024 0.178 Di-2-ethylhexyl phthalate concentration (μM) 0.000 0.042
0.127 0.381 1.143 3.42810.284 30.852 92.556 277.66
7 833.000 Growth (Absorbance at 650nm) 0.245 0.235 0.235 0.234
0.238 0.223 0.2130.188 0.159 0.119 0.055 2,5-Dichlorophenol concentration (μM) 0.000 0.169 0.508 1.
524 4.572 13.716 41.148 123.444 370.333 1111.000 3333.000 Growth (Abso
rbance at 650nm) 0.252 0.264 0.260 0.262 0.235 0.2
38 0.2300.193 0.036 0.029 0.024 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.248 0.262 0.257 0.265
0.254 0.246 0.2180.191 0.135 0.032 0.045 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.568 61.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.252 0.266 0.248 0.247
0.262 0.233 0.2270.174 0.035 0.034 0.033 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.280 0.271 0.275 0.269
0.249 0.185 0.0120.010 0.010 0.014 0.042 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.281 0.265 0.234 0.165
0.279 0.282 0.2650.259 0.166 0.036 0.013 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.286 0.294 0.331 0.383
0.279 0.361 0.2700.150 0.041 0.035 0.025 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.285 0.270 0.265 0.201
0.110 0.027 0.0260.023 0.022 0.035 0.046 Potassium
dichlomate concentration (μM) 0.000 0.002 0.005 0.015 0.0
45 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absor
bance at 650nm) 0.281 0.267 0.546 0.276 0.275 0.73
6 0.2710.264 0.259 0.473 0.218 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.323 0.454 0.311 0.303
0.476 0.288 0.2870.290 0.275 0.273 0.240

[0013]

(3) Chemical substance sensitivity of Glysomyces rutagersensis IFO14488 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.066 0.065 0.069 0.068
0.047 0.042 0.0380.032 0.026 0.021 0.018 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.041 0.038 0.039 0.042
0.042 0.044 0.0420.040 0.025 0.025 0.024 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.076 0.070 0.076 0.069
0.061 0.058 0.0470.029 0.003 -0.015 -0.157 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.058 0.053 0.052 0.051
0.050 0.043 0.0370.029 0.025 0.026 0.036 Malathion concentration (μM) 0.000 0.034 0.102 0.305 0.914
2.741 8.222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.059 0.052 0.053 0.052
0.054 0.053 0.0540.051 0.048 0.045 0.006 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.044 0.059 0.059 0.061
0.059 0.054 0.0500.047 0.038 0.032 0.025 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.049 0.046 0.046 0.050
0.049 0.048 0.0490.066 0.073 0.090 -0.082 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.052 0.049 0.049 0.046
0.044 0.038 0.0290.027 0.028 0.028 0.030 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.053 0.053 0.053 0.050
0.049 0.047 0.0420.027 0.027 0.029 0.133 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.066 0.061 0.059 0.056
0.055 0.054 0.0480.047 0.043 0.036 0.030 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.055 0.053 0.058 0.058
0.054 0.049 0.0450.042 0.035 0.026 0.019 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.053 0.051 0.054 0.055
0.054 0.054 0.0510.049 0.052 0.046 0.041 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorba
(nce at 650nm) 0.057 0.050 0.050 0.051 0.050 0.047
0.0440.042 0.032 0.025 0.030 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.045 0.043 0.042 0.037
0.033 0.033 0.0330.031 0.029 0.029 0.027 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.060 0.058 0.062 0.064
0.059 0.055 0.0520.042 0.030 0.026 0.058Di-2-ethy
lhexyl phthalate concentration (μM) 0.000 0.042 0.127 0.38
1 1.143 3.42810.284 30.852 92.556 277.667 833.000 Growth (Absorbance at 650nm) 0.052 0.052 0.053 0.051
0.053 0.051 0.0500.039 0.036 0.022 0.020 2,5-Dichlorophenol concentration (μM) 0.000 0.169 0.508 1.
524 4.572 13.716 41.148 123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.062 0.059 0.061 0.060
0.059 0.056 0.0540.044 0.030 0.024 0.0202,4-Dichl
orophenoxy acetic acid concentration (μM) 0.000 0.339 1.01
6 3.048 9.14427.432 82.296 246.889 740.667 2222.00
0 6666.000 Growth (Absorbance at 650nm) 0.062 0.058 0.063 0.061
0.057 0.055 0.0520.050 0.033 0.022 0.026 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.054 0.050 0.054 0.054
0.053 0.052 0.0510.050 0.048 0.036 0.021 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.056 0.054 0.052 0.052
0.048 0.045 0.0310.021 0.019 0.023 0.069 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.111 12.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.049 0.047 0.049 0.047
0.047 0.045 0.0440.040 0.040 0.041 0.035 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.053 0.048 0.049 0.047
0.044 0.042 0.0340.028 0.023 0.022 0.025 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.056 0.053 0.053 0.050
0.039 0.033 0.0310.029 0.031 0.030 0.046 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.136 0.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.041 0.036 0.041 0.043
0.043 0.043 0.0440.042 0.040 0.037 0.037 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.014 0.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.044 0.045 0.049 0.049
0.049 0.052 0.0500.044 0.034 0.026 0.020

[0014]

[Table 4] (4) Aureobacterium esteraromatium IFO 37
Chemical sensitivity of 52 Methylmercury chloride Concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.091 0.107 0.110 0.093
0.043 0.023 0.0220.023 0.022 0.021 0.018 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.098 0.106 0.116 0.112
0.116 0.111 0.1120.114 0.110 0.084 0.016 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.095 0.093 0.113 0.104
0.099 0.099 0.0900.091 0.069 0.008 -0.108 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.105 0.037 0.044 0.044
0.042 0.038 0.0300.018 0.014 0.023 0.032 Malathion
Concentration (μM) 0.000 0.034 0.102 0.305 0.914 2.741 8.
222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.101 0.098 0.122 0.120
0.119 0.113 0.1050.100 0.088 0.068 -0.001 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.100 0.099 0.121 0.114
0.107 0.105 0.1010.098 0.085 0.045 0.023 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.103 0.110 0.131 0.132
0.126 0.099 0.0770.093 0.103 0.111 -0.059 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.095 0.098 0.116 0.115
0.093 0.054 0.0220.021 0.019 0.017 0.021 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.089 0.098 0.107 0.107
0.103 0.089 0.0560.039 0.030 0.023 0.056 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.106 0.111 0.121 0.118
0.118 0.118 0.1010.096 0.081 0.068 0.066 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.111 12.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.096 0.097 0.118 0.117
0.113 0.110 0.1070.099 0.087 0.077 0.126 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.097 0.089 0.120 0.117
0.112 0.104 0.0890.085 0.065 0.049 0.032 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.107 0.102 0.121 0.120
0.122 0.119 0.1160.105 0.074 0.024 0.036 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.090 0.094 0.119 0.119
0.112 0.105 0.0930.085 0.046 0.020 0.020 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.088 0.092 0.123 0.123
0.121 0.111 0.0950.080 0.019 0.018 0.052 Di-2-ethylhexyl phthalate concentration (μM) 0.000 0.042
0.127 0.381 1.143 3.42810.284 30.852 92.556 277.66
7 833.000 growth (Absorbance at 650nm) 0.087 0.069 0.
122 0.119 0.121 0.117 0.1090.096 0.075 0.052 0.048 2,5-Dichlorophenol concentration (μM) 0.000 0.169 0.508 1.
524 4.572 13.71641.148 123.444 370.333 1111.000 33
33.000 Growth (Absorbance at 650nm) 0.097 0.103 0.129 0.127
0.128 0.122 0.1130.086 0.020 0.017 0.019 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.093 0.103 0.124 0.122
0.124 0.124 0.1160.100 0.075 0.018 0.027 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.097 0.107 0.122 0.123
0.119 0.121 0.1200.112 0.084 0.044 0.015 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.102 0.090 0.076 0.111
0.108 0.095 0.0150.016 0.013 0.022 0.101 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.098 0.094 0.122 0.120
0.119 0.119 0.1110.107 0.100 0.088 0.065 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.105 0.097 0.124 0.122
0.123 0.109 0.0820.040 0.020 0.017 0.016 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.098 0.095 0.115 0.110
0.081 0.021 0.0200.020 0.020 0.023 0.033 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.105 0.101 0.121 0.122
0.124 0.117 0.1180.120 0.116 0.109 0.088 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.100 0.096 0.107 0.110
0.109 0.103 0.1000.085 0.056 0.029 0.019

[0015]

(5) Chemical sensitivity of Comamonas Testosteroni IAM1048 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.420 0.419 0.417 0.381
0.080 0.059 0.0560.056 0.057 0.048 0.039 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.316 0.302 0.314 0.313
0.303 0.302 0.2930.285 0.279 0.178 -0.005 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.417 0.412 0.435
0.420 0.425 0.413 0.3900.335 0.248 0.118 -0.084 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.401 0.406 0.401 0.394
0.373 0.268 0.1550.074 0.056 0.051 0.072 Malathion concentration (μM) 0.000 0.034 0.102 0.305 0.914
2.741 8.222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.341 0.341 0.346 0.319
0.314 0.300 0.2880.270 0.234 0.141 -0.101 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.321 0.302 0.312 0.315
0.319 0.321 0.3050.302 0.269 0.156 0.000 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.323 0.312 0.316 0.313
0.302 0.267 0.2400.258 0.405 0.750 1.157 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.393 0.404 0.401 0.400
0.379 0.337 0.2820.158 0.039 0.035 0.037 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.391 0.562 0.456 0.411
0.418 0.361 0.2670.089 0.046 0.035 0.089 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.399 0.406 0.412 0.39
8 0.402 0.386 0.3660.355 0.345 0.284 0.140 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.416 0.425 0.445 0.443
0.432 0.430 0.4250.409 0.386 0.306 -0.108 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.327 0.314 0.326 0.313
0.304 0.294 0.2590.168 -0.085 -0.543 -1.517 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.316 0.304 0.314 0.311
0.304 0.303 0.2840.257 0.165 0.041 0.003 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.307 0.294 0.302 0.294
0.287 0.297 0.2860.262 0.201 0.068 -0.269 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.413 0.420 0.425 0.428
0.423 0.388 0.3530.223 0.054 0.049 0.157 Di-2-ethylhexyl phthalate concentration (μM) 0.000 0.042
0.127 0.381 1.143 3.42810.284 30.852 92.556 277.66
7 833.000 Growth (Absorbance at 650nm) 0.432 0.414 0.440 0.437
0.438 0.426 0.4360.417 0.110 0.021 -0.143 2,5-Dichlorophenol concentration (μM) 0.000 0.169 0.508 1.
524 4.572 13.71641.148 123.444 370.333 1111.000 33
33.000 Growth (Absorbance at 650nm) 0.436 0.442 0.447 0.449
0.454 0.430 0.1340.083 0.054 0.049 0.040 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.427 0.430 0.438 0.443
0.446 0.434 0.4210.390 0.099 0.044 0.052 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.437 0.428 0.444 0.446
0.442 0.427 0.4330.429 0.430 0.382 0.077 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.403 0.387 0.412 0.408
0.388 0.364 0.2930.216 0.182 0.142 0.022 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.407 0.407 0.403 0.436
0.416 0.399 0.3830.365 0.333 0.242 0.129 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.404 0.401 0.426 0.416
0.391 0.362 0.2530.191 0.083 0.037 0.047 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.411 0.398 0.397 0.390
0.313 0.063 0.0500.052 0.049 0.041 0.077 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.323 0.303 0.322 0.318
0.311 0.303 0.2960.296 0.289 0.273 0.226 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.321 0.315 0.328 0.327
0.317 0.304 0.2820.280 0.237 0.197 0.118

[0016]

(6) Chemical sensitivity of Pseudomonas synxantha IFO 3913 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.719 0.716 0.728 0.733
0.736 0.476 0.0700.060 0.056 0.057 0.046 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.667 0.686 0.673 0.673
0.673 0.666 0.6620.666 0.659 0.542 0.051 2-Aminoanthracene concentration (μM) 0.000 0.169 0.508 1.5
24 4.572 13.716 41.148123.444 370.333 1111.000 333
3.000 growth (Absorbance at 650nm) 0.784 0.785 0.804 0.794
0.805 0.803 0.7690.723 0.682 0.548 0.150 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.748 0.773 0.767 0.776
0.769 0.761 0.7610.729 0.351 0.077 0.057 Malathion concentration (μM) 0.000 0.034 0.102 0.305 0.914
2.741 8.222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.717 0.714 0.708 0.696
0.695 0.680 0.6880.706 0.778 0.751 0.180 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.624 0.649 0.646 0.615
0.594 0.541 0.4220.298 0.203 0.116 0.053 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.655 0.657 0.647 0.660
0.644 0.606 0.5930.597 0.571 0.451 -0.032 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.770 0.775 0.774 0.763
0.756 0.726 0.7110.692 0.708 0.731 0.158 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.701 0.712 0.726 0.723
0.723 0.714 0.7110.730 0.700 0.382 0.149 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.746 0.754 0.768 0.748
0.753 0.741 0.7180.725 0.747 0.845 0.630 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.782 0.802 0.814 0.819 0.8
03 0.806 0.7930.775 0.773 0.774 0.558 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (Absorbance at 650nm) 0.680 0.691 0.679 0.669
0.659 0.638 0.5550.440 0.308 0.221 0.154 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.695 0.720 0.712 0.697
0.676 0.668 0.6690.692 0.760 0.808 0.261 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.677 0.704 0.696 0.698
0.683 0.676 0.6480.592 0.529 0.471 0.381 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.787 0.805 0.812 0.817
0.821 0.807 0.8210.836 0.573 0.686 0.455Di-2-ethy
lhexyl phthalate concentration (μM) 0.000 0.042 0.127 0.38
1 1.143 3.42810.284 30.852 92.556 277.667 833.000 Growth (Absorbance at 650nm) 0.784 0.757 0.792 0.794
0.783 0.772 0.7730.772 0.788 0.783 0.582 2,5-Dichlorophenol concentration (μM) 0.000 0.169 0.508 1.
524 4.572 13.71641.148 123.444 370.333 1111.000 33
33.000 Growth (Absorbance at 650nm) 0.788 0.805 0.810 0.818
0.822 0.824 0.8300.852 0.767 0.075 0.024 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.761 0.773 0.785 0.797
0.801 0.795 0.8010.835 0.906 0.945 0.045 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.765 0.752 0.776 0.784
0.785 0.779 0.7890.789 0.784 0.765 0.740 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.758 0.762 0.775 0.763
0.737 0.708 0.7260.621 0.484 0.420 0.289 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.776 0.773 0.786 0.778
0.762 0.735 0.7150.680 0.663 0.615 0.520 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.769 0.773 0.774 0.771
0.773 0.756 0.7370.703 0.652 0.624 0.354 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.750 0.759 0.763 0.759
0.736 0.701 0.6570.562 0.044 0.033 0.081 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.644 0.664 0.668 0.659
0.651 0.631 0.6360.634 0.630 0.602 0.501 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.624 0.631 0.621 0.636
0.635 0.622 0.6210.641 0.672 0.738 0.418

[0017]

(7) Chemical substance sensitivity of Staphylococcus aureus IFO 3060 Methylmercury chloride concentration (μM) 0.000 0.034 0.10
2 0.305 0.914 2.7418.222 24.667 74.000 222.000 66
6.000 Growth (Absorbance at 650nm) 0.184 0.198 0.200 0.189
0.007 0.006 0.0060.007 0.008 0.007 0.005 Nickel chloride concentration (μM) 0.000 0.169 0.508 1.524
4.572 13.716 41.148123.444 370.333 1111.000 3333.
000 Growth (Absorbance at 650nm) 0.237 0.232 0.226 0.227
0.224 0.220 0.2160.216 0.208 0.111 0.0072-Aminoan
thracene concentration (μM) 0.000 0.169 0.508 1.524 4.572
13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.174 0.181 0.176 0.173
0.170 0.168 0.1670.155 0.124 0.121 0.033 Tributyltin chloride concentration (μM) 0.000 0.003 0.010
0.030 0.091 0.2720.815 2.444 7.333 22.000 66.000 Growth (Absorbance at 650nm) 0.154 0.177 0.185 0.183
0.160 0.109 0.0850.045 0.020 0.010 0.029 Malathion concentration (μM) 0.000 0.034 0.102 0.305 0.914
2.741 8.222 24.66774.000 222.000 666.000 Growth (Absorbance at 650nm) 0.235 0.226 0.223 0.219
0.216 0.211 0.2150.222 0.231 0.226 0.063 Phenol concentration (μM) 0.000 0.847 2.540 7.620 22.861 6
8.584 205.753 617.2591851.778 5555.333 16666.000 Growth (Absorbance at 650nm) 0.256 0.192 0.199 0.198
0.197 0.193 0.1930.190 0.180 0.159 0.043 Lindane concentration (μM) 0.000 0.339 1.016 3.048 9.144 2
7.432 82.296 246.889740.667 2222.000 6666.000 Growth (Absorbance at 650nm) 0.177 0.095 0.085 0.080
0.075 0.071 0.0800.074 0.106 0.055 -0.230 Pentachlorophenol concentration (μM) 0.000 0.034 0.102 0.3
05 0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.146 0.148 0.142 0.129
0.107 0.090 0.0730.041 0.007 0.002 0.004 Trp-P-2 (Acetate) concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.189 0.200 0.199 0.187
0.176 0.143 0.0840.041 0.017 0.012 0.085 4-Nitroquinoline-N-oxide concentration (μM) 0.000 0.001 0.
003 0.008 0.023 0.0700.210 0.630 1.889 5.667 17.00
0 Growth (Absorbance at 650nm) 0.182 0.185 0.185 0.183
0.181 0.177 0.1850.188 0.191 0.170 0.104 Benzo (a) pyrene concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.162 0.164 0.156 0.158
0.138 0.114 0.0940.076 0.068 0.062 0.094 Paraquat concentration (μM) 0.000 0.051 0.152 0.457 1.372
4.115 12.346 37.037111.111 333.333 1000.000 Growth (A
(bsorbance at 650nm) 0.249 0.241 0.218 0.217 0.217
0.211 0.1960.177 0.174 0.175 0.161 Maneb concentration (μM) 0.000 0.017 0.051 0.152 0.457 1.3
70 4.111 12.333 37.000111.000 333.000 Growth (Absorbance at 650nm) 0.232 0.230 0.225 0.226
0.223 0.215 0.2160.212 0.187 0.134 0.013 Cadmium chloride concentration (μM) 0.000 0.017 0.051 0.15
2 0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.237 0.233 0.221 0.215
0.203 0.164 0.1590.123 0.090 0.076 0.029 Bis-phenol-A concentration (μM) 0.000 0.169 0.508 1.524 4.
572 13.716 41.148123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.159 0.158 0.156 0.156
0.158 0.163 0.1620.109 0.003 0.003 0.085 Di-2-ethylhexyl phthalate concentration (μM) 0.000 0.042
0.127 0.381 1.143 3.42810.284 30.852 92.556 277.66
7 833.000 Growth (Absorbance at 650nm) 0.158 0.157 0.159 0.152
0.153 0.150 0.1510.153 0.143 0.080 0.0692,5-Dichl
orophenol concentration (μM) 0.000 0.169 0.508 1.524 4.572
13.71641.148 123.444 370.333 1111.000 3333.000 Growth (Absorbance at 650nm) 0.156 0.162 0.158 0.157
0.156 0.152 0.0860.053 0.029 0.001 0.002 2,4-Dichlorophenoxy acetic acid concentration (μM) 0.000
0.339 1.016 3.048 9.14427.432 82.296 246.889 740.6
67 2222.000 6666.000 Growth (Absorbance at 650nm) 0.158 0.159 0.155 0.159
0.159 0.153 0.1460.116 0.066 0.005 0.011 Formaldehyde concentration (μM) 0.000 0.085 0.254 0.762 2.
285 6.856 20.56861.704 185.111 555.333 1666.000 Growth (Absorbance at 650nm) 0.160 0.159 0.163 0.164
0.162 0.161 0.1590.161 0.160 0.156 0.147 p-Nonylphenol concentration (μM) 0.000 0.034 0.102 0.305
0.914 2.741 8.22224.667 74.000 222.000 666.000 Growth (Absorbance at 650nm) 0.158 0.165 0.164 0.167
0.143 0.156 0.0700.068 0.052 0.055 0.160 Sodium Arsenite concentration (μM) 0.000 0.017 0.051 0.152
0.457 1.370 4.11112.333 37.000 111.000 333.000 Growth (Absorbance at 650nm) 0.148 0.148 0.148 0.147
0.149 0.143 0.1410.131 0.094 0.094 0.070 Thiuram concentration (μM) 0.000 0.017 0.051 0.152 0.457
1.370 4.111 12.33337.000 111.000 333.000 Growth (Absorbance at 650nm) 0.145 0.150 0.150 0.153
0.151 0.146 0.1340.092 0.029 0.006 0.004 2,4,5-Trichlorophenol concentration (μM) 0.000 0.169 0.508
1.524 4.572 13.71641.148 123.444 370.333 1111.000
3333.000 Growth (Absorbance at 650nm) 0.152 0.155 0.161 0.154
0.109 0.034 0.0060.005 0.005 0.006 0.000 Potassium dichlomate concentration (μM) 0.000 0.002 0.005
0.015 0.045 0.1360.407 1.222 3.667 11.000 33.000 Growth (Absorbance at 650nm) 0.241 0.227 0.225 0.224
0.220 0.216 0.2190.222 0.218 0.207 0.188 Triphenyltin chloride concentration (μM) 0.000 0.000 0.001
0.002 0.005 0.0140.041 0.122 0.367 1.100 3.300 Growth (Absorbance at 650nm) 0.256 0.274 0.260 0.257
0.255 0.254 0.2480.232 0.129 0.097 0.024

As the indicator microorganism, it is preferable to select many different types of microorganisms that are susceptible to growth inhibition. As such a microorganism, an essential indicator microorganism for each chemical substance is preferably selected. In order to select the essential indicator microorganism, first, for each chemical substance, the minimum concentration that inhibits the growth of Escherichia coli is measured, and a plurality of microorganisms that are inhibited from growing at a concentration lower than the minimum inhibitory concentration are selected (primary microorganisms). screening). Next, the microorganisms selected in this primary screening are grown in the presence of different concentrations of indicator chemicals, and the characteristics of growth inhibition by the chemicals are observed. From these results, the microorganism showing the highest sensitivity to the chemical substance among the microorganisms selected in the primary screening is selected as an essential indicator microorganism. Of course, as the indicator microorganism, various microorganisms that are more susceptible to growth inhibition than Escherichia coli can be used as the indicator microorganism in addition to the essential indicator microorganism.

As understood from the results shown in Tables 1 to 7, Methylmercury chloride, Nickel chloride, 2-A
Marker chemicals such as minoanthracene, Malathion, Pheno1, and Lindane, except for Methylmercury chloride, clearly have significant inhibition on the indicator microorganism. However, Methyl
Regarding mercury chloride and tributyltin chloride, remarkable growth inhibition was observed in several microorganisms other than the indicator microorganism. Methylmercury chloride shows almost the same inhibitory properties against all the indicator microorganisms, but it is understood that it shows the characteristics of Methylmercury chloride because no other 24 chemicals are found. it can. Tributyltin chloride is inhibited only by three kinds of microorganisms, so that it can be understood that the growth spectrum also shows the characteristics of Tributyltin chloride. From the above, it is clear that the growth characteristics or the production spectrum (microorganism / chemical substance concentration data) of the indicator microorganism for the above chemical substance reflects the characteristics of the chemical substance.

Pentachlorophenol, Trp-P-2 (Acetate), 4-N
itroquinoline-N-oxide, Benzo (a) pyrene, Paraquat, Mane
b, Cadmium chloride, because of the presence of microorganisms sensitive to these compounds, the growth spectrum,
It is characteristic. For example, Pentachloropheno
Acetobacter pasteurianusIF03129 showed sensitivity to l. This microorganism was originally selected as a microorganism showing sensitivity to Nickel chloride, but Pentachlorophe
It can be understood that nol is also sensitive. However, due to the different sensitivities of other microorganisms to Pentachlorophenol and Nickel chloride, their growth spectra clearly indicate that both chemicals are different.
Similarly, Trp-P-2 (Acetate), 4-Nitroquinoline-N-oxide,
Benzo (a) pyrene, Paraquat, Maneb, and Cadmiumchloride also have indicator microorganisms showing susceptibility, but they can be distinguished from each other because their spectra are different.

Bis-phenol-A, Di-2-ethyl hexylphthalat
e, 2,5-Dichlorophenol, 2,4-Dichlorophenoxy acid, Form
aldehyde, p-Nonylphenol, Sodium Arsenite, Thiuram, 2,
4,5-Trichlorophenol, Potassium dichlomate, Triphenyl
For tin chloride, there were no significantly susceptible microorganisms. Of these compounds, 2,5-DichIoroph
eno1,2,4-Dichlorophenoxy acetic acid, 2,4,5-Trichlo
With the exception of rophenol, the microbial susceptibility to these compounds is different, resulting in characteristic growth spectra of chemicals. 2,5-Dichlorophenol, 2,4-Dic
The growth spectra of hlorophenoxy acetic acid, 2,4,5-trichlorophenol were similar. However, since these compounds are similar compounds, it is presumed that their toxicity is also similar, and it can be considered that the growth spectra are similar. Conversely, it can be considered as an example showing that the method of the present invention is effective for expressing the toxicity of a chemical substance.

The method of the present invention includes the step of growing a plurality of indicator microorganisms in the presence of the sample chemical. The growth conditions in this case are the same conditions as when the indicator chemical substance was grown in the presence of the indicator microorganism described above. Further, the method of the present invention includes a measuring step of measuring the relationship between the concentration of the microorganism and the concentration of the sample compound in the growing step. In the method of the present invention, the microorganism / chemical substance concentration data obtained in the measurement step is converted into the indicator microorganism /
A collation step for collating with the indicator chemical substance concentration data is included. In this collation step, when indicator microorganism / indicator chemical substance concentration data that is the same as or similar to the sample chemical substance is detected, the sample compound substance is microbiologically identical or similar to the indicator chemical substance in the detected data. Identified as similar. Sample chemicals that can be used in the present invention include:
It does not need to be a high-purity chemical substance, and may be environmental water containing the chemical substance, contaminated water contaminated with the chemical substance, or the like. Each of the above steps can be automatically performed. The collation of the data obtained in the measurement process with the data stored in the data bank can be automatically performed using a computer.

[0023]

According to the present invention, a sample chemical substance can be microbiologically identified. That is, the type and structure of the chemical substance and the toxicity of the chemical substance to microorganisms can be known by a microbiological technique. Such a method has never been known before and was first obtained by the present invention, and its industrial significance is great.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Katsuhide Fujita 2-9-16-107 Fujimidaira, Hamura City, Tokyo (72) Inventor Satomi Gumi 1-13-24, Uesugi, Aoba-ku, Sendai, Miyagi Prefecture 201 (72) Inventor Sachiko Takahashi 1-3-1 Higashi, Tsukuba, Ibaraki Pref., National Institute of Advanced Industrial Science and Technology (72) Inventor Akiyoshi Matsuyama 1-14-14-304 Sengen, Tsukuba, Ibaraki F term (reference 4B063 QA01 QA18 QQ61 QR74 QS24 QX01

Claims (1)

[Claims] 1. A microorganism growing step in which a plurality of predetermined indicator microorganisms are grown by changing the concentration of the sample chemical substance in the presence of the sample chemical substance, and the concentration of the sample chemical substance and the indicator microorganism in the microorganism growing step. A measuring step of measuring the relationship between the concentration, and the relationship between the indicator microorganism concentration and the sample chemical concentration obtained in the measuring process, the indicator microorganism concentration and the indicator chemical concentration stored in advance in a data bank. A microbiological identification method for a chemical substance, comprising a collation step of collating with data indicating the relationship of