Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates

Toxicity to microorganisms

Link to relevant study record(s)

Reference

Endpoint:

toxicity to microorganisms, other

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Data of read across substance

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: As mention below

Principles of method if other than guideline:

WoE report was prepared based on two toxicity studies of microorganisms for the test chemical :

GLP compliance:

not specified

Specific details on test material used for the study:

- Name of test material : Reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates- Substance type: Organic- Physical state: Solid- Physical Appearance: Violet powder

Analytical monitoring:

not specified

Vehicle:

not specified

Test organisms (species):

other: Agrobacterium radiobacter, A. tumefaciens, Bradyrhizobium japonicum, E. coli, Erwinia atroseptica, E. herbicola, E. uredovora, P.fluorescence, P. phaseolicola, P. syringae, Rhizobium trifolii, Xanthomonas malvacearum, X. phaseoli, X. stewartii

Details on inoculum:

1 and 2 - Laboratory culture: Test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary).- Name and location of sewage treatment plant where inoculum was collected: No data available- Method of cultivation: A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth.- Preparation of inoculum for exposure: Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C.- Pretreatment: No data available- Initial biomass concentration: No data availableIn 2nd study another gram positive bacteria were used: Gram – positive bacteria: Corynebacterium fascians, Corynebacterium flaccumfaciens, Corynebacterium michiganense, Corynebacterium oortii, Stapylococcus aureus, Micrococcus luteus, Bacillus subtilis and Bacillus thuringiensis Berliner subp. kurstaki

Test type:

not specified

Water media type:

not specified

Total exposure duration:

24 h

Remarks on exposure duration:

After 24 hrs of incubation, colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth.

Test temperature:

21 ± 1 °C

Nominal and measured concentrations:

0.6918 mg/l (1 µM)

Details on test conditions:

1. TEST SYSTEM - Test vessel: Petri dishes- Material, size, headspace, fill volume: The diameter of Petri dishes was 90 mm.EFFECT PARAMETERS MEASURED (with observation intervals if applicable): After 24 hrs of incubation, colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. - Test concentrations: 0.6918 mg/l (1 µM)

Reference substance (positive control):

not specified

Key result

Duration:

24 h

Dose descriptor:

NOEC

Effect conc.:

0.692 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: In 1st study

Key result

Duration:

24 h

Dose descriptor:

LOEC

Effect conc.:

0.62 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

growth inhibition

Remarks on result:

other: 1 and 2nd study

1. Table:Microbiological activity of synthetic dyes towards bacteria.

Test organism	Growth inhibition (in %)
Agrobacterium radiobacter	11
Agrobacterium tumefaciens	11
Bradyrhizobium japonicum	28
Escherichia coli	15
Erwinia atroseptica	8
Erwinia herbicola	0
Erwinia uredovora	13
Pseudomonas fluorescence	6
Pseudomonas phaseolicola	13
Pseudomonas syringae	0
Rhizobium trifolii	11
Xanthomonas malvacearum	6
Xanthomonas phaseoli	19
X. stewartii	9

Study 2: Table: Microbiological activity of test chemical toward bacterias.

Test organism	Growth inhibition (in %)
Corynebacterium fascians	18
Corynebacterium flaccumfaciens	19
Corynebacterium michiganense	17
Corynebacterium oortii	13
Stapylococcus aureus	30
Micrococcus luteus	19
Bacillus subtilis	20
Bacillus thuringiensisBerliner subp.kurstaki	17

Validity criteria fulfilled:

not specified

Conclusions:

1. Based on no effect on growth inhibition of gram negative test organisms, the NOEC value of test material was determine to be 0.6918 mg/l for Erwinia herbicola and Pseudomonas syringae. Whereas the growth inhibition was observed on the other organisms so LOEC was 0.6918 mg/l for remaining 12 bacteria.2. Based on growth inhibition of gram positive test organisms, the LOEC value of test chemical was determine to be 0.6918 mg/l. Based on above experimental studies it was concluded that the reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates shows low effect at 0.6194 mg/l.

Executive summary:

Summarized results of the data available for the structurally and functional similar read across chemicals study has been reviewed to determine the toxicity of the test reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates on the growth of microorganisms. The studies are as mentioned below:

 

Determination of toxicity of test material on the growth of 14 gram negative microorganisms. Inhibitory activity of test chemical was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM). 14 different gram negative bact. were used for the study was Agrobacterium radiobacter, Agrobacterium tumefaciens, Bradyrhizobium japonicum, Escherichia coli, Erwinia atroseptica,Erwinia herbicola, Erwinia uredovora, Pseudomonas fluorescence, Pseudomonas phaseolicola, Pseudomonas syringae , Rhizobium trifolii, Xanthomonas malvacearum, Xanthomonas phaseoli and X. stewartii. These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on no effect on growth inhibition of gram negative test organisms, the NOEC value of test material was determine to be 0.6918 mg/l for Erwinia herbicola and Pseudomonas syringae. Whereas the growth inhibition was observed on the other organisms so LOEC was 0.6918 mg/l for remaining 12 bacteria.

Determination of toxicity of test chemical on the growth of 8 gram positive microorganisms. Inhibitory activity of test chemical was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM).

8 different gram positive bact. were used for the study-

Gram – positive bacteria:

Corynebacterium fascians

Corynebacterium flaccumfaciens

Corynebacterium michiganense

Corynebacterium oortii

Stapylococcus aureus

Micrococcus luteus

Bacillus subtilis and

Bacillus thuringiensis Berliner subp. kurstaki

These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on growth inhibition of gram positive test organisms, the LOEC value of test substance was determine to be 0.6918 mg/l.

Based on above experimental studies it was concluded that the reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates shows low effect at 0.6194 mg/l.

Description of key information

1. Based on no effect on growth inhibition of gram negative test organisms, the NOEC value of test material was determine to be 0.6918 mg/l for Erwinia herbicola and Pseudomonas syringae. Whereas the growth inhibition was observed on the other organisms so LOEC was 0.6918 mg/l for remaining 12 bacteria.

2. Based on growth inhibition of gram positive test organisms, the LOEC value of test chemical was determine to be 0.6918 mg/l.      

Based on above experimental studies it was concluded that the reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates shows low effect at 0.6194 mg/l.

Key value for chemical safety assessment

EC10 or NOEC for microorganisms:

0.692 mg/L

Additional information

Summarized results of the data available for the structurally and functional similar read across chemicals study has been reviewed to determine the toxicity of the test reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates on the growth of microorganisms. The studies are as mentioned below:

 

Determination of toxicity of test material on the growth of 14 gram negative microorganisms. Inhibitory activity of test chemical was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM). 14 different gram negative bact. were used for the study was Agrobacterium radiobacter, Agrobacterium tumefaciens, Bradyrhizobium japonicum, Escherichia coli, Erwinia atroseptica,Erwinia herbicola, Erwinia uredovora, Pseudomonas fluorescence, Pseudomonas phaseolicola, Pseudomonas syringae , Rhizobium trifolii, Xanthomonas malvacearum, Xanthomonas phaseoli and X. stewartii. These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on no effect on growth inhibition of gram negative test organisms, the NOEC value of test material was determine to be 0.6918 mg/l for Erwinia herbicola and Pseudomonas syringae. Whereas the growth inhibition was observed on the other organisms so LOEC was 0.6918 mg/l for remaining 12 bacteria.

Determination of toxicity of test chemical on the growth of 8 gram positive microorganisms. Inhibitory activity of test chemical was determined by the traditional agar gel method. The conc. of test chemical used for the study was 0.6918 mg/l (1 µM).

8 different gram positive bact. were used for the study-

Gram – positive bacteria:

Corynebacterium fascians

Corynebacterium flaccumfaciens

Corynebacterium michiganense

Corynebacterium oortii

Stapylococcus aureus

Micrococcus luteus

Bacillus subtilis and

Bacillus thuringiensis Berliner subp. kurstaki

These test organisms were taken from the collection of the Plant Protection Institute, Hungarian Academy of Sciences (Budapest, Hungary). The bacteria were maintained on Nutrient Agar (Oxoid CM3) completed with vitamins pyridoxine. HCl, thiamine. HCl, riboflavine and nicotinamide at 1, 10, 1 and 20 mg/l concentrations. Bacterial suspensions for screening were prepared by washing cells with sterile tap water containing 0.3% peptone from slants of 20 h old cultures grown at (21±1) °C. A layer of 5 mm depth of inoculated medium was dispensed into Petri dishes of 90 mm diameter. Filter paper discs of 5 mm diameter were impregnated with 1 µM solution of test compounds and placed centrally on the surface of the agar plate. Growth inhibition zones were measured after 24 h incubation at 21 ± 1 °C. After incubation colony diameters were determined and growth inhibition was calculated in percentage of dye free colony growth. Each determination was run in quadruplicate. Based on growth inhibition of gram positive test organisms, the LOEC value of test substance was determine to be 0.6918 mg/l.

Based on above experimental studies it was concluded that the reaction mass of Benzenamine, N,N-dimethyl- , molybdate, tungstate & phosphates shows low effect at 0.6194 mg/l.