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Toxicity to soil macroorganisms except arthropods

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Endpoint:
toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Study was performed according to ISO 10872 guideline and is well documented.
Qualifier:
according to
Guideline:
other: ISO 10872 "Water Quality – Determination of the toxic effect of sediment and soil samples on growth, fertility and reproduction of Caenorhabditis elegans"
Deviations:
no
GLP compliance:
yes
Remarks:
The study has no formal GLP certificate. However, the study was conducted at a GLP-certified laboratory, staff was appropriately trained and standard operating procedures were followed in terms of experimental performance and raw data documentation.
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- The test item was dissolved in a sufficient amount of deionised water to prepare a stock solution. This stock solution was diluted with deionised water to obtain one test item solution (= dosage solution) for each test item concentration.
- Method of mixing into soil (if used): The test item was incorporated into the soil by thoroughly mixing in the respective test item solution for approx. 3 to 5 minutes.
- This amount of water/test item-mixture served also to adjust the final soil moisture of 40% of the dry weight.
Test organisms (species):
Caenorhabditis elegans
Animal group:
nematods
Details on test organisms:
The nematodes used in this test, Caenorhabditis elegans were taken from a breeding culture kept at ECT Oekotoxikologie GmbH.
The nematodes were breed on agar plates at 20 ± 2°C. The cultures were fed with Escherichia coli (OP50 uracil-poor strain) and moistened on demand with deionised water.
The nematodes inserted in the test were synchronised juveniles of the first larval stage
Study type:
laboratory study
Substrate type:
natural soil
Limit test:
no
Total exposure duration:
96 h
Test temperature:
20 +- 0.5 °C
pH:
soil pH-value: 5.7 – 5.8
Moisture:
soil moisture: 37.6 – 40.0% of the dry weight
Details on test conditions:
TEST SYSTEM
- Test container (material, size): a well with an area of 3.9 cm² and a volume of 2 to 4 mL in a 12 well multiwell plate
- Amount of soil or substrate: 0.5g fresh weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 2
- No. of replicates per control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Geographic location: Germany
- Soil name: LUFA 2.2
- Sampling depth (cm): 20
- Soil texture (if natural soil): loamy sand
- Organic carbon (%): 2.29
- Maximum water holding capacity (in % dry weigth): 49.7
- CEC: 10 mval/100g
- Sampling date: December 14, 2007
- Vegetation; uncultivated

OTHER TEST CONDITIONS
- Photoperiod: permanent dark

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) : reproduction

Nominal and measured concentrations:
nominal added concentrations: control, 10.0, 31.6, 56.2, 75.0, 100, 237, 562 and 1000 mg H3BO3/kg dry weight soil,
corresponding to control, 1.75, 5.5, 9.8, 13.1, 17.5, 41.4, 98.2 and 174.8 mg B/kg dw
Reference substance (positive control):
no
Duration:
96 h
Dose descriptor:
EC10
Effect conc.:
86.7 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
number of juvenile nematodes
Reported statistics and error estimates:
Data were checked for normality by Shapiro-Wilk´s test on normal distribution and for homogeneity by Cochran’s test. Treatment means were compared by ANOVA and Williams test (p ≤ 0.05, one-sided) and tested for statistically significant differences compared to the control. Probit analysis using Linear Max. Likelihood Regression was used to calculate the EC50,EC20 and EC10 for reproduction.

96 h LOEC for reproduction: 174.8 mg B/kg soil dw (82% inhibition)

96 h NOEC for reproduction: 98.2 mg B/kg soil dw (18.6% inhibition)

96 h EC50 for reproduction: 130.6 mg B/kg soil dw

96 h LOEC for mortality: 98.2 mg B/kg soil dw (55% mortality)

96 h NOEC for mortality: 98.2 mg B/kg soil dw

 

Concentration

[mg test item/kg soil (dw)]

Number of Juveniles[mean ± sd]  Number of Juveniles[% of control]
 Control 1250.0 +-334.1 
 10.0 951.0 +- 190.9 76.1 
 31.6 834.5 +- 402.3  66.8 
 56.2 1040.5 +- 573.5  83.2 
 75.0 1141.0 +- 335.2  91.3 
 100 1163.0 +- 258.8  93.0 
 237 1203.5 +- 171.8  96.3 
 562 1017 +- 18.4 81.4 
 1000 225.0 +- 18.4  18.0 
Validity criteria fulfilled:
yes
Conclusions:
This reliable study was performed according to the ISO 10872 guideline and is well documented. A 96-h EC10 value for reproduction of nematods of 86.7 mg B/ kg soil dw is obtained and used for PNEC derivation.
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed according to OECD 222 (2004) and ISO 11268-2 Part 2 (1998) guidelines and is well documented.
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Deviations:
no
GLP compliance:
yes
Remarks:
The study has no formal GLP certificate. However, the study was conducted at a GLP-certified laboratory, staff was appropriately trained and standard operating procedures were followed in terms of experimental performance and raw data documentation.
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- The test item was dissolved in a sufficient amount of deionised water to prepare a stock solution. This stock solution was diluted with deionised water to obtain one test item solution (= dosage solution) for each test item concentration.
- Method of mixing into soil (if used): The test item was incorporated into the soil by thoroughly mixing in the respective test item solution for approx. 5 minutes.

Test organisms (species):
Eisenia fetida
Animal group:
annelids
Details on test organisms:
The earthworms used in this test, Eisenia fetida (Lumbricidae, Earthworms) are kept at ECT Oekotoxikologie GmbH since February 1994.
Only adult worms (with clitellum) with a fresh weight (FW) between 300 and 600 mg were used. The worms were at least two months, but not more than one year old, however the age of individuals did not differ by more than four weeks. Animals which do fulfil these requirements were taken from a synchronised culture. The worms selected for the test were acclimatised in artificial soil under test conditions at least 24 hours before starting the test.
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
28 d
Remarks:
56 days for reproduction endpoint
Test temperature:
19-21 °C
pH:
soil pH-value: 5.9
Moisture:
soil moisture: 40-60% of WHCmax
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Test vessels were Bellaplast containers consisting of inert (non-toxic) plastic (Polystyrol). The test vessels have a bottom area of 11 x 15.5 cm (=170.5 cm²) and a height of 6 cm. The test vessels were covered by a transparent lid.
- Amount of soil or substrate: 500 g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 2
- No. of replicates per control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- artificial soil based on OECD guideline 207 (1984). Percentages refer to dry weight:
- 5% Sphagnum peat (no visible plant remains; finely ground and air-dried)
- 20% Kaolin-Clay (kaolinite content > 30%)
- 74% Quartz sand (fine sand content with particles between 50 and 200 µm higher than 50%)
- ca. 1% Calcium carbonate (CaCO3)
- The moisture content was adjusted to 40 – 60% of WHCmax using deionised water.

OTHER TEST CONDITIONS
- Photoperiod: 16 : 8 h light/dark with a constant light intensity at the substrate surface of 400 – 800 lx
- Feeding: The adult worms were fed with finely ground cow manure (free of growth promoters, nematicides or similar veterinary pharmaceuticals,
). Before starting the experiment an amount of 5 g food per 500 g soil dry weight was mixed into the artificial soil to be used in the test. After the start of the experiment food was first provided one day after application of the test item and introduction of the adult worms. Thereafter, the adult worms were fed weekly during the first 4 weeks of the test. An amount of 5 g moistened with 10 mL deionised water was spread on the soil surface of each test container. If food remains uneaten the ration was reduced on demand. After removing of the adults on day 28 further 5 g of food was mixed with the artificial soil of each test vessel.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality: mean number of dead adult worms as a percentage of the initial number at the start of the test for each concentration and for the control.
- Biomass: loss/increase in the mean biomass (i.e. the fresh mass of organisms) 28 days after application expressed as a percentage of the initial fresh mass of organisms at the start of the test for each concentration and for the control.
- Reproduction: mean number of juveniles per test vessel 56 days after application.

Nominal and measured concentrations:
nominal added concentrations: control, 75, 100, 133, 178, 237, 316, 422, 562, 750 and 1000 mg H3BO3/kg dry weight soil,
corresponding to control, 13.1, 17.5, 23.2, 41.4, 55.2, 73.8, 98.2, 131.1 and 174.8 mg B/kg dw
Reference substance (positive control):
no
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 174.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
added B
Basis for effect:
mortality
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
>= 174.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
added B
Basis for effect:
growth
Remarks:
biomass production
Duration:
56 d
Dose descriptor:
EC10
Effect conc.:
70.1 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
added B
Basis for effect:
reproduction
Remarks:
number of juveniles
Reported statistics and error estimates:
Mortality and Biomass: No statistical analysis was performed.

Reproduction: Data were checked for normality by Shapiro-Wilk´s test on normal distribution and for homogeneity by Cochran’s test. Treatment means were compared by ANOVA followed by Dunnett’s t-test (p ≤ 0.05, according to the guideline two-sided for biomass and one-sided for reproduction) and tested for statistically significant differences compared to the control. Logit analysis (2-parameter-logistic function) using linear maximum likelihood regression was used to calculate the EC50, EC20 and EC10 for reproduction.

56 d EC50 for reproduction: 84.6 mg B/kg soil dw

56 d EC20 for reproduction: 75.2 mg B/kg soil dw

56 d NOEC for reproduction: 73.8 mg B/kg soil dw (16.6% inhibition)

56 d LOEC for reproduction: 98.2 mg B/kg soil dw (85.3% inhibition)

 

Concentration

[mg test item/kg soil (dw)]

Number of dead adultsDay 28 Mortality (%)Day 28
 Control 0 0
 75 0 0
 100 2 10
133 0 0
 178 0 0
 237 0
 316 0 0
 422 0 0
 562 0 0
 750 0 0
 1000 0 0

 

Concentration

[mg test item/kg soil (dw)]

Number of Juveniles[mean ± sd]  Number of Juveniles[% of control]
 Control 357.3 +- 45.2   100.0 
 75 320.0 +- 7.1 89.6 
 100 431.0 +- 80.6  120.6 
133 454.0 +- 15.6  127.1 
 178 406.5 +- 19.1  113.8 
 237 446.0 +- 50.9  124.8 
 316 418.0 +- 75.0  117.0 
 422 298.0 +- 21.2 83.4 
 562 52.5 +- 9.2  14.7 
 750  1.0 +- 1.4 0.3 
 1000  0.0 +- 0.0 0.0 

 

Concentration

[mg test item/kg soil (dw)]

Biomass (mg fresh weight)Day 0 Biomass (mg fresh weight)Day 28  % of initial weightDay 28
 Control 418.7±65.7 624.3±106.3  149.1
 75 427.0±66.0 660.9±137.1  154.8
 100 416.2±75.2 591.7±227.2  142.2
133 418.2±58.3 672.4±166.4  160.8
 178 421.0±73.9 656.7±98.1 156.0 
 237 433.2±65.7 680.3±116.3 157.0
 316 424.4±45.5 708.6±99.2 167.0
 422 416.5±80.0 655.6±107.4 157.4
 562 416.3±71.4 639.8±125.9 153.7
 750 431.1±61.2 615.6±89.3 142.8
 1000 423.6±62.0 484.8±56.4 114.4
Validity criteria fulfilled:
yes
Conclusions:
This reliable study was performed according to the OECD 222 guideline and is well documented. A 56 days EC10 value for reproduction of Eisenia fetida of 70.1 mg B/ kg soil dw is obtained and used for PNEC derivation.
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed according to OECD 220 (2004) guideline and is well documented.
Qualifier:
according to
Guideline:
OECD Guideline 220 (Enchytraeid Reproduction Test)
Deviations:
no
GLP compliance:
yes
Remarks:
The study has no formal GLP certificate. However, the study was conducted at a GLP-certified laboratory, staff was appropriately trained and standard operating procedures were followed in terms of experimental performance and raw data documentation.
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- The test item was dissolved in a sufficient amount of deionised water to prepare a stock solution. This stock solution was diluted with deionised water to obtain one test item solution (= dosage solution) for each test item concentration.
- Method of mixing into soil (if used): The test item was incorporated into the soil by thoroughly mixing in the respective test item
solution for approx. 5 minutes.
- This amount of water/test item-mixture served also to adjust the final soil moisture of 40-60% of WHCmax.
Test organisms (species):
other: Enchytraeus crypticus
Animal group:
other: Enchytraeidae
Details on test organisms:
Enchytraeus crypticus (Enchytraeidae, potworms), has been cultured at ECT Oekotoxikologie GmbH since September 2003.
The worms used in the tests were adult (with eggs, i.e. white spots, in the clitellum region).
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
28 d
Test temperature:
19-21 °C
pH:
soil pH-value: 5.9
Moisture:
soil moisture: 40.0-60.0% of the WHCmax
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Test vessels were glass containers (able to be closed tightly) of about 200 mL capacity and with a diameter of about 5 cm.
- Amount of soil or substrate: 20g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 2
- No. of replicates per control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- artificial soil based on OECD guideline 207 (1984). Percentages refer to dry weight:
- 10% Sphagnum peat (no visible plant remains; finely ground and air-dried)
- 20% Kaolin-Clay (kaolinite content > 30%)
- 69.6% Quartz sand (fine sand content with particles between 50 and 200 µm higher than 50%)
- 0.4% Calcium carbonate (CaCO3)
- The moisture content was adjusted to 40 – 60% of WHCmax using deionised water.

OTHER TEST CONDITIONS
- Photoperiod: 16 : 8 h light/dark with a constant light intensity at the substrate surface of 400 – 800 lx
- Feeding: The enchytraeids were fed with about 50 mg of ground oats per test vessel at the beginning of the test (mixed with the test substrate after
application of the test item). The organisms were fed an additional 25 mg ground oats once per week until day 21.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality: mean number of dead adult worms as a percentage of the initial number at the start of the test for each concentration and for the control. (Validity criteria for mortality not fulfilled)
- Reproduction: mean number of juveniles per test vessel 28 days after application.


Nominal and measured concentrations:
nominal added concentrations: control, 1.0, 10, 32, 100, 133, 178, 237, 316, 422, 562, 750 and 1000 mg H3BO3/kg dry weight soil,
corresponding to control, 0.17, 1.75, 5.6, 17.5, 23.2, 31.1, 41.4, 55.2, 73.8, 98.2, 131.1 and 174.8 mg B/kg dw
Reference substance (positive control):
no
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
22.6 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
added B
Basis for effect:
reproduction
Remarks:
number of juveniles
Reported statistics and error estimates:
Data were checked for normality by Shapiro-Wilk´s test on normal distribution and for homogeneity by Cochran’s test. Treatment means were compared by ANOVA and Welsh-t test (p ≤ 0.05, one-sided) and tested for statistically significant differences compared to the control.
Probit analysis using Linear Max. Likelihood Regression was used to calculate the EC50, EC20 and EC10 for reproduction.

28 d NOEC for reproduction: 23.2 mg B/kg soil dw (5.2% inhibition)

28 d LOEC for reproduction: 31.1 mg B/kg soil dw (33.1% inhibition)

28 d EC50 for reproduction: 38.5 mg B/kg soil dw

28 d EC20 for reproduction: 27.1 mg B/kg soil dw

28 d NOEC for mortality: >174.8 mg B/kg soil dw

 

Concentration

[mg test item/kg soil (dw)]

Number of Juveniles[mean ± sd]  Number of Juveniles[% of control]
 Control 628.7± 92.8  
 1.0 638.0± 148.5 101.5
 10 677.5± 87.0 107.8 
 32 778.0± 48.1 123.8
 100 690.0± 200.8 109.8 
 133 596.0± 15.6 94.8 
 178 420.5± 43.1 66.9 
 237 228.0± 29.7 36.3
 316 162.0± 8.5 25.8 
 422  47.5± 4.9  7.6
 562 17.5± 9.2 2.8
 750 6.0± 1.4 1.0
 1000 1.0± 1.4 0.2
Validity criteria fulfilled:
yes
Conclusions:
This reliable study was performed according to the OECD 220 guideline and is well documented. A 28-d EC10 value for reproduction of Enchytraeus crypticus of 22.6 mg B/ kg soil dw is obtained and used for PNEC derivation.
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study was performed according to OECD 220 (2004) guideline and is well documented.
Qualifier:
according to
Guideline:
OECD Guideline 220 (Enchytraeid Reproduction Test)
Deviations:
no
GLP compliance:
yes
Remarks:
The study has no formal GLP certificate. However, the study was conducted at a GLP-certified laboratory, staff was appropriately trained and standard operating procedures were followed in terms of experimental performance and raw data documentation.
Analytical monitoring:
no
Vehicle:
no
Details on preparation and application of test substrate:
- The test item was dissolved in a sufficient amount of deionised water to prepare a stock solution. This stock solution was diluted with deionised water to obtain one test item solution (= dosage solution) for each test item concentration.
- Method of mixing into soil (if used): The test item was incorporated into the soil by thoroughly mixing in the respective test item
solution for approx. 5 minutes.
- This amount of water/test item-mixture served also to adjust the final soil moisture of 40-60% of WHCmax.
Test organisms (species):
other: Enchytraeus luxuriosus
Animal group:
other: Enchytraeidae
Details on test organisms:
Enchytraeus luxuriosus (Enchytraeidae, potworms), has been cultured at ECT Oekotoxikologie GmbH since September 2003.
The worms used in the tests were adult (with eggs, i.e. white spots, in the clitellum region).
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
28 d
Test temperature:
18-21 °C
pH:
soil pH-value: 6.5
Moisture:
soil moisture: 40.0-60.0% of the WHCmax
Details on test conditions:
TEST SYSTEM
- Test container (material, size): Test vessels were glass containers (able to be closed tightly) of about 200 mL capacity and with a diameter of about 5 cm.
- Amount of soil or substrate: 20g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 2
- No. of replicates per control: 6

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- artificial soil based on OECD guideline 207 (1984). Percentages refer to dry weight:
- 10% Sphagnum peat (no visible plant remains; finely ground and air-dried)
- 20% Kaolin-Clay (kaolinite content > 30%)
- 69.6% Quartz sand (fine sand content with particles between 50 and 200 µm higher than 50%)
- 0.4% Calcium carbonate (CaCO3)
- The moisture content was adjusted to 40 – 60% of WHCmax using deionised water.

OTHER TEST CONDITIONS
- Photoperiod: 16 : 8 h light/dark with a constant light intensity at the substrate surface of 400 – 800 lx
- Feeding: The enchytraeids were fed with about 50 mg of ground oats per test vessel at the beginning of the test (mixed with the test substrate after application of the test item). The organisms were fed an additional 25 mg ground oats once per week until day 21.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality: mean number of dead adult worms as a percentage of the initial number at the start of the test for each concentration and for the control. (Validity criteria for mortality not fulfilled)
- Reproduction: mean number of juveniles per test vessel 28 days after application.


Nominal and measured concentrations:
nominal added concentrations: control, 10, 32, 56, 75, 100, 133, 178, 237, 316, 422, 562 and 1000 mg H3BO3/kg dry weight soil,
corresponding to control, 1.75, 5.6, 9.8, 13.1, 17.5, 23.2, 31.1, 41.4, 55.2, 73.8, 98.2 and 174.8 mg B/kg dw
Reference substance (positive control):
no
Duration:
28 d
Dose descriptor:
EC10
Effect conc.:
17 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
added B
Basis for effect:
reproduction
Remarks:
number of juveniles
Reported statistics and error estimates:
Data were checked for normality by Shapiro-Wilk´s test on normal distribution and for homogeneity by Cochran’s test. Treatment means were compared by ANOVA and Welsh-t test (p ≤ 0.05, one-sided) and tested for statistically significant differences compared to the control.
Probit analysis using Linear Max. Likelihood Regression was used to calculate the EC50, EC20 and EC10 for reproduction.

28 d NOEC for reproduction: 23.2 mg B/kg soil dw (26.2% inhibition)

28 d LOEC for reproduction: 31.1 mg B/kg soil dw (39.7% inhibition)

28 d EC50 for reproduction: 39.9 mg B/kg soil dw

28 d EC20 for reproduction: 22.7 mg B/kg soil dw

28 d NOEC for mortality: >174.8 mg B/kg soil dw

 

Concentration

[mg test item/kg soil (dw)]

Number of Juveniles[mean ± sd]  Number of Juveniles[% of control]
 Control 937.5± 191.9  
 10 875.5± 119.5 93.4
 32 1024.5± 102.5 109.3 
 56 1214.5± 211.4 129.5
 75 1268.0± 414.4 135.3
100 876.0± 77.8 93.4
133 701.0± 1.4 74.8
178 565.0± 41.0 60.3
237 400.0± 11.3 42.7 
316 437.0± 171.1 46.6
422 112.5± 50.2 12.0
 562 43.5± 54.5 1.0
 1000 38.0± 31.1 4.1
Validity criteria fulfilled:
yes
Conclusions:
This reliable study was performed according to the OECD 220 guideline and is well documented. A 28-d EC10 value for reproduction of Enchytraeus luxuriosus of 17 mg B/ kg soil dw is obtained and is used fpr PNEC derivation.
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: according to THE OECD guideline or national guideline (Environment Canada), standard test and selection criteria fulfilled.
Qualifier:
according to
Guideline:
OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))
Deviations:
not specified
Qualifier:
according to
Guideline:
other: Environment Canada (2002)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
not specified
Details on preparation and application of test substrate:
Test compound added in solution to soil. Soil moisture was 35% (based on wet weight). Tests started after a 1 day equilibration period.
Test organisms (species):
Eisenia sp.
Animal group:
annelids
Details on test organisms:
Eisenia andrei
- fully developed clitellum
- aged 2-12 months, synchronized
- individual wet weight 250-600 mg
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
56 d
Test temperature:
20°C
pH:
6.09
Moisture:
35% (based on wet weight)
Details on test conditions:
TEST SYSTEM
Environment Canada guideline:
- Test container (material, size): 500 ml
- Amount of soil or substrate: 350 ml (200 g)
- No. of organisms per container (treatment): 2
- No. of replicates per treatment group: 10
- No. of replicates per control: 10

OECD guideline:
- Test container (material, size): 1-2 L
- Amount of soil or substrate: 500-600 g dry weight
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8

SOURCE AND PROPERTIES OF ARTIFICIAL SOIL
- Ingredients: 70% silica sand, 20% kaolinite clay, 10% spaghnum peat, 1-3g calcium carbonate per kg soil
- Soil texture
- % sand: 77.3
- % silt: 7.8
- % clay: 14.9
- Total carbon (%): 4.46
- Organic matter (%): 9
- pH water (1:2): 6.09
- eCEC: 18.5 cmol c/kg
- background B concentration: 3.5 mg B/kg dw

OTHER TEST CONDITIONS
- Photoperiod: 16h light, 8h dark

Nominal and measured concentrations:
0-10-16-30-50-100-300-560-1000 mg boric acid/kg dw,
corresponding to 0-1.75-2.8-5.25-8.75-17.5-52.5-98-175 mg B/kg dw.
Reference substance (positive control):
not specified
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
5.2 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching failure)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
98 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
77.9 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
25.7 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
98 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
136.2 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: Environment Canada guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
52.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching failure)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
NOEC
Effect conc.:
98 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
75.7 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
59.4 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
90.4 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: OECD guideline
Duration:
56 d
Dose descriptor:
EC50
Effect conc.:
107.6 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(cocoon production)
Remarks on result:
other: OECD guideline
Details on results:
No significant effect on mortality up to the largest dose tested (175 mg B/kg dw).
Reported statistics and error estimates:
ECx values and their associated 95% confidence limits for the chronic tests were analysed by using either a linear or four nonlinear regression models (i.e., logistic, Gompertz, exponential and logistic with hormoesis). If data showed heteroscedascity among treatments, data were weighed with the inverse of the variance of each treatment.
Validity criteria fulfilled:
yes
Conclusions:
This study followed the OECD guideline or a national guideline (Environment Canada)
The standard test and selection criteria were fulfilled.
Reliable NOEC values are used for PNEC derivation.
Endpoint:
toxicity to soil macroorganisms except arthropods: long-term
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: according to national guideline (Environment Canada), standard test and selection criteria fulfilled
Qualifier:
according to
Guideline:
other: Environment Canada (2002)
Deviations:
not specified
GLP compliance:
not specified
Analytical monitoring:
not specified
Vehicle:
not specified
Details on preparation and application of test substrate:
Test compound added in solution to soil. Soil moisture was 35% (based on wet weight). Tests started after a 1 day equilibration period.
Test organisms (species):
Eisenia sp.
Animal group:
annelids
Details on test organisms:
Eisenia andrei
- fully developed clitellum
- aged 2-12 months
- individual wet weight 250-600 mg
Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
no
Total exposure duration:
63 d
Post exposure observation period:
not reported
Test temperature:
20°C
pH:
6.09
Moisture:
35% (based on wet weight)
Details on test conditions:
TEST SYSTEM
- Test container (material, size): 500 ml
- Amount of soil or substrate: 350 ml (200 g)
- No. of organisms per container (treatment): 2
- No. of replicates per treatment group: 10
- No. of replicates per control: 10


SOURCE AND PROPERTIES OF ARTIFICIAL SOIL
- Ingredients: 70% silica sand, 20% kaolinite clay, 10% spaghnum peat, 1-3g calcium carbonate per kg soil
- Soil texture
- % sand: 77.3
- % silt: 7.8
- % clay: 14.9
- Total carbon (%): 4.46
- Organic matter (%): 9
- pH water (1:2): 6.09
- eCEC: 18.5 cmol c/kg
- background B concentration: 3.5 mg B/kg dw

OTHER TEST CONDITIONS
- Photoperiod: 16h light, 8h dark
Nominal and measured concentrations:
0-7-14-28-56-113-225-450-900-1800-3600 mg boric acid/kg dw,
corresponding to 0-1.2-2.5-4.9-9.8-19.8-39.4-78.8-315-630 mg B/kg dw
Reference substance (positive control):
not specified
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
315 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
mortality
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
39.4 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
19.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching failure)
Remarks on result:
other: asynchronous cultures
Duration:
28 d
Dose descriptor:
NOEC
Effect conc.:
315 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
mortality
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
NOEC
Effect conc.:
>= 78.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching failure)
Remarks on result:
other: synchronous culture
Duration:
28 d
Dose descriptor:
LC50
Effect conc.:
589.2 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
mortality
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
77.4 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
95.5 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: asynchronous cultures
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
120.2 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: asynchronous cultures
Duration:
28 d
Dose descriptor:
EC50
Effect conc.:
620 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
mortality
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
99.4 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(juvenile production)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
165.8 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
growth
Remarks:
(juvenile dry weight)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
86.3 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching success)
Remarks on result:
other: synchronous culture
Duration:
63 d
Dose descriptor:
EC50
Effect conc.:
172.1 mg/kg soil dw
Nominal / measured:
nominal
Conc. based on:
element
Remarks:
B
Basis for effect:
reproduction
Remarks:
(hatching failure)
Remarks on result:
other: synchronous culture
Details on results:
nominal added dosis; background concentration reported
Reported statistics and error estimates:
ECx values and their associated 95% confidence limits for the chronic tests were analysed by using either a linear or four nonlinear regression models (i.e., logistic, Gompertz, exponential and logistic with hormoesis). If data showed heteroscedascity among treatments, data were weighed with the inverse of the variance of each treatment.
NOEC values were estimated using ANOVA, Dunnett's and Fisher's protected LSD (least significant difference) pairwise comparison test.
Validity criteria fulfilled:
yes
Conclusions:
This study followed the Environment Canada guideline.
The standard test and selection criteria were fulfilled. Reliable NOEC values are used for PNEC derivation.

Description of key information

Key value for chemical safety assessment

Additional information

In total 36 individual high quality NOEC/EC10 values (for 9 different species) are selected for the PNEC derivation, ranging from 5.2 mg B/kg dw for Eisenia andrei growth to 315 mg B/kg for Eisenia andrei survival (Stantec Consulting & Aquaterra Environmental Consulting, 2003).

 

Boric acid and borates have been registered as pesticides effective against a number of terrestrial insects, for example cockroaches, termites and ants. However the studies that establish efficacy as a pesticide are not relevant for use in describing ecotoxicological responses from boron present in soil because these pesticide studies typically involve use of a specially formulated diet or bait or treated article.

For example, Durmuş and Büyükgüzel (2008) reported effects on survival and development of larvae of the Greater Wax Moth, Galleria mellonella, when fed artificial diets containing 0.2 % sodium tetraborate. No effects were observed when fed 0.1 % sodium tetraborate. Yang et al. (2000) reported reduction in survival and reproduction of thefruit fly Anastrepha suspensa when fed 0.1 to 0.5 % sodium tetraborate. Since the diets were not soil-based (as might occur for an earthworm), such studies cannot be relied upon for derivation of a PNECsoil. The critical boron concentrations in food (approx. 0.1 % or 1000 mg B/kg) are orders of magnitude above the soil effects concentrations of boron for invertebrates (5-300 mg B/kg). Because boron does not magnify through the food-chain and soil to plant bioaccumulation factors for boron derived from tests performed in real soils are generally < 100 and internal boron concentrations in plants are largely controlled by plant-specific requirements for this essential element, it can be concluded that the secondary poisoning of insects through plant material is not critical for boron.