Calcium cyanamide

Toxicity to terrestrial arthropods

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline available

Principles of method if other than guideline:

Adult beetles were placed in dishes of moist soil in which Perlka had been applied to the surface at a rate of 450 kg/ha. The survival of beetles was monitored over a 5-day period, in relation to control beetles.

GLP compliance:

not specified

Application method:

soil

Specific details on test material used for the study:

Batch J-26/1-90

Analytical monitoring:

no

Details on sampling:

no analytical monitoring

Vehicle:

no

Details on preparation and application of test substrate:

The test substrate was loam soil obtained from Southampton University botanical gardens; this had not previously been treated with agrochemicals. The soil was sieved and its pH and organic matter determined. The pH using Corning 245 pH meter. The organic matter content was determined by igniting dried soil in a furnace at 425°C for 8 hours (result: 3.70 % by weight).
For the test, a 2 cm-deep layer of the soil was lightly compacted into the base of a plastic pot (9cm diameter and 4.5 cm depth). The moisture content of the soil used was adjusted to approximately 70 % of its capacity, to do this, the volume of the water was needed to fully saturate a sub-sample was determined and then 70 % of that amount was added to the soil in each pot.
The test product was applied at an application rate of 450 kg/ha. The granules were shaken evenly over the soil surface in each pot.

Test organisms (species):

Bembidion lampros

Animal group:

Coleoptera (soil-dwelling carabid beetle)

Details on test organisms:

Adult beetles were collected from arable field boundaries on the Leckford estate, Stockbridge, Hampshire, U.K., during April 1990. Pitfall traps were set at the boundary edges and emptied at 3-day intervals. The captured arthropods were transported back to the laboratory beetles of the species bembidion lampros were separated from the other captured arthropodes using an aspirator. They were placed, in groups of ten, into Petri dishes lined with moist filter paper. They were provided with moistened dog-food pellets as food, which were replaced every three days in order to prevent fungal growth developing. The beetles were stored in a controlled environment room at a temperature of approximately 15°C and under 16 h day length for four days prior to testing. The beetles were assumed to be mixed sexes and of approximately the same age.

Study type:

laboratory study

Total exposure duration:

5 d

Post exposure observation period:

none

Test temperature:

15 °C +/- 2°C

pH (if soil or dung study):

6.65

Humidity:

70 %

Photoperiod and lighting:

16 h light

Details on test conditions:

Five test insects were placed onto the treated soil surface in each pot and a ventilated clear plastic lid was fitted. Five replicate pots were used for each treatment and five untreated pots were used as a control. A crumbled moistened dog-food pellet was provided in each pot as food. The pots were placed in a controlled environment room operating at 15 °C with 2°C range and a 16 h photoperiod. Three days after the start of the experiment, the food supply was replenished and each pot lightly moistened.
The condition of the beetles was assessed at one, six and 24 h after being introduced into the pots. Thereafter they were assessed daily for a further four days. In each assessment, the condition of the beetles was recorded as being: live, affected, moribund, dead, missing.
For the 5-day assessment the pots were emptied into plastic trays in order to recover all of the individuals that may have burrowed into the soil.

Nominal and measured concentrations:

Nominal: 450 kg/ha

Reference substance (positive control):

no

Key result

Duration:

5 d

Dose descriptor:

NOEC

Effect conc.:

> 450 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: no adverse effects observed

Details on results:

When applied to the soil surface, Perlka was not found to be toxic to foraging Bembidion lampros. During the test, some of the beetles were not found as they had burrowed into the soil or down the edge of the pots. Most of these missing beetles were recovered in the final assessment. A few (2-3 of 25 beetles) were not recovered from both the control and the Perlka treatments. It was assessed that these had died and had been cannibalised by the other beetles. Therefore, beetle mortality after 5 days was 16 % in the Perlka treatment, or 8.7 % once corrected for the control mortality of 8 % (using Abbot's formula).
It is not possible to determine to what extent the test beetles were exposed to the active ingredient of this granular product. Although the test substrate was wetted to 70 % of its capacity prior to the bioassay, it is not known how much of the surface-applied granules will have gone into solution. Since the granules appeared to remain intact on the soil surface throughout the five-day test, a possible route of exposure for the beetles could have been through ingestion of the product. However, since the beetles are only 3-4 mm in length and the smallest granules were approximately 1 mm in diameter, this is unlikely to have occurred.

Results with reference substance (positive control):

no positive control

Reported statistics and error estimates:

correction with Abbot's formula

Validity criteria fulfilled:

not specified

Conclusions:

Perlka was not found to be toxic to Bembidion lampros up to a soil surface concentration of 450 kg/ha.

Executive summary:

A laboratory study was carried out to determine the potential hazard of Perlka granules to the carabid beetle Bembidion lampros (herbst), when applied to a soil surface. Adult beetles were placed in dishes of moist soil in which Perlka had been applied to the surface at a rate of 450 kg/ha. The survival of beetles was monitored over a 5-day period, in relation to control beetles. Under these conditions, Perlka was not found to be toxic to Bembidion lampros and it was considered unlikely that the product would be hazardous to these beetles under field conditions.

Reference 2

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

40 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

53.2 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.4 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

19.1 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

22.662 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: recalculated by Competent Authority

Key result

Duration:

28 d

Dose descriptor:

EC10

Effect conc.:

1.515 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: recalculated by Competent Authority

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

0.4 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: recalculated by Competent Authority

Details on results:

Effects on mortality:
The LC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression to be 53.2 mg a.s./kg soil (dw) (95% confidence limits: 48.0 – 58.9 mg a.s./kg).

Statistical analysis (Fisher’s Exact test; 1-sided, p ≤ 0.05) revealed a significant difference concerning the mortality between the control and the concentrations of 60.00, 80.00 and 100.00 mg a.s./kg soil (dw).
Therefore, the LOEC Mortality as 60.00 mg a.s./kg soil (dw) and accordingly the NOEC Mortality was determined to be 40.00 mg a.s./kg soil (dw).
For detailed results please refer to table Table 1 in the attached document.

Effects on reproduction:
The ANOVA and the Williams’ test (1-sided, p ≤ 0.05) showed a significant difference concerning the number of juveniles between the control and the concentrations of 1.26, 4.00, 12.46, 40.00, 60.00, 80.00 and 100.00 mg a.s./kg soil (dw).
Therefore, the NOEC Reproduction was determined to be 0.40 mg a.s./kg soil (dw) and accordingly the LOEC Reproduction as 1.26 mg a.s./kg soil (dw).
For detailed results please refer to table Table 2 in the attached document.

The results for reproduction were statistically re-evaluated by the Competent Authority (please refer to Reported statistics and error estimetes).
The values re-calculated by the Competent Authority are:
EC50 = 22.662 mg a.s./kg soil (dw) (95% confidence limits: 14.087 – 37.194 mg a.s./kg dw)
EC10 = 1.515 mg a.s./kg soil (dw) (95% confidence limits: 0.229 – 3.555 mg a.s./kg dw)
NOEC (reproduction) 0.40 mg a.s./kg soil (dw)

Results with reference substance (positive control):

In the reference test with the toxic standard, the number of juveniles was significantly (Student-t test, 1-sided, p <= 0.05) reduced by 73.9% in comparison to the control.
Therefore, the observed effects of the toxic standard show the high sensitivity of the test system.

Reported statistics and error estimates:

Mortality: The LC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression. The NOEC (mortality) was determined by pair-wise comparisons between treatment and control by Fisher’s Exact test (p ≤ 0.05, one-sided).
Reproduction: The EC50 value for reproduction was calculated by Probit analysis using Linear Max. Likelihood Regression. For determination of the NOEC (reproduction), data were checked for normality by R/s test procedure 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.

The results for reproduction were statistically re-evaluated by the Competent Authority:
• ECX values by Weibull analysis using linear weighted regression;
• NOEC by Kolmogorov-Smirnov-Test, Cochran's Test Procedure and Williams Multiple Sequential t-Test Procedure.
The resulting effect values have to be considered relevant for the effect assessment and the risk assessment.

Validity criteria fulfilled:

yes

Conclusions:

In a dose-response test with the collembolan species Folsomia candida, the NOEC (mortality) was calculated to be 53.2 mg a.s./kg soil dw. The NOEC (reproduction) was determined to be 0.4 mg a.s./kg soil dw. The NOEC and EC10 (reproduction) recalculated by the Competent Authority were determined to be 0.40 mg a.s./kg soil dw and 1.515 mg/kg soil dw, respectively.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

Materials and methods:

The test was performed according to the guideline ISO 11267 (1999): "Soil Quality – Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants".

Deviations: Due to a malfunction of the pH-electrode, the pH value could not be determined at the start of the test (i. e. after incorporation of the test solution). However, the pH value determined after preparation of the batch of artificial soil (before incorporation of the test solution) used in this study was 5.7 (mean of three measurements) which is in the range required by the guideline.

In all concentrations of the test item tested and in the control the observed pH value at the end of the test was slightly lower than the recommended range (5.2 and 5.4 instead of 5.5).

Nevertheless, the observed survival and reproduction rates of the collembolans did fulfil the validity criteria and were in the historical range of the results of ECT laboratory. Therefore, the deviations can be regarded as minor and have no impact on the outcome of the study.

Results and discussion:

All validity criteria according to ISO 11267 are fulfilled and the study can be considered as valid:

Average mortality of the adult test animals in the control (required ≤ 20%): yes

Average reproduction rate per vessel in the control (required: at least 100 juveniles per vessel): yes

Coefficient of variation of reproduction in the control (required: ≤ 30%): yes

The results are based on the active substance content of the test item and show a clear dose-response relationship.

LC50 (mortality) 53.20 mg a.s./kg soil (dw)

NOEC (mortality) 40.00 mg a.s./kg soil (dw)

EC50 (reproduction) 19.10 mg a.s./kg soil (dw) / 22.662 mg a.s./kg soil (dw) *

EC10 (reproduction) 4.20 mg a.s./kg soil (dw) / 1.515 mg a.s./kg soil (dw) *

NOEC (reproduction) 0.40 mg a.s./kg soil (dw) / 0.40 mg a.s./kg soil (dw) *

* Values recalculated by the Competent Authority are given in bold font.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 3

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

4 wk

Dose descriptor:

LOEC

Effect conc.:

< 20 other: Kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Details on results:

A statistically significant reduction (Dunnett-test, p = 0.05) in numbers of emerging F-1 generation beetles was observed at the test substance rate (3 ± 2 beetles) relative to the control (881 ± 98 beetles). Reproduction (parasitism success) was reduced by 99.7 %. Reproduction was reduced by 80.4 % (173 ± 114 beetles) in the toxic standard.

Results with reference substance (positive control):

Reproduction was reduced by 80.4 % (173 ± 114 beetles) in the toxic standard.

Reported statistics and error estimates:

Dunnett-test, p = 0.05

The effects on beetle emergence and reproduction (parasitism success) of the Staphylinid beetle, Aleochara biliniata exposed to a 51.1 % (w/w) aqueous solution of cyanamide in the laboratory:

Test substance	51.1 % (w/w) aqueous solution of cyanamide
Test organism	Aleochara biliniata
Exposure	28 d, sand
% Reduction in reproduction* (20 kg pure cyanamide/ha)	99.7 %

Validity criteria fulfilled:

yes

Conclusions:

Reproduction (parasitism success) was reduced by 99.7%. It was concluded that a 51.1 % (w/w) aqueous solution of cyanamide was harmful to the rove beetle Aleochara bilineata when applied to sand surface at 39.14 kg product/ha, which is equivalent to the recommended maximum field rate of 20 kg a.i./ha in 400-600 L water/ha.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according GLP and following the IOBC/WPRS guidelines for testing the effects of pesticides on beneficials (Moreth & Naton, 1992).

Emergence of the F-1 generation of Aleochara biliniata beetles and the parasitism success on Delia pupae were evaluated following a 4-week exposure period. Adult beetles were placed in beakers containing sand substrate. SKW Cyanamid L500 (a 51.1 % (w/w) aqueous solution of cyanamide) was applied to the sand surface at 97.85 g product/L before introduction of the test organisms. This rate was equivalent to the proposed field rate of 39.14 kg SKW Cyanamid L500/ha. The application volume was equivalent to 400 l/ha. This equates to 20 kg pure active ingredient cyanamide/ha. An untreated control and a toxic standard (437 g Dimethoate/ha) were also tested.

A statistically significant reduction (Dunnett-test, p = 0.05) in numbers of emerging F-1 generation beetles was observed at the test substance rate (3 ± 2 beetles) relative to the control (881 ± 98 beetles). Reproduction (parasitism success) was reduced by 99.7 %. Reproduction was reduced by 80.4 % (173 ± 114 beetles) in the toxic standard.

It can be concluded that a 51.1 % (w/w) aqueous solution of cyanamide is harmful to rove beetles, Aleochara bilineata, when applied to sand surface at 39.14 kg product/ha, equivalent to the recommended maximum field rate of 20 kg a.i./ha in 400 -600 L water/ha.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 4

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Reference substance (positive control):

yes

Duration:

7 d

Dose descriptor:

LR50

Effect conc.:

445.6 g/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

196 g/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

100 g/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

339 g/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

LOEC

Effect conc.:

173 g/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

Mortality in the control and toxic standard groups was 5 % and 89.5 %, respectively.
The reproduction of surviving mites was significantly reduced at 587 g test substance/ha (equivalent to 300 g cyanamide/ha). However at a rate of 1018 g test substance/ha (equivalent to 520 g pure cyanamide/ha), reproductive performance was not significantly different from the control. Therefore, the result observed in this study was not dose related

Results with reference substance (positive control):

Mortality in the toxic standard group was 89.5% (in the control group, 5%)

Reported statistics and error estimates:

The test groups were significantly different from the control, examined by Dunnet-test, α=0.05

Effects on the predatory mite, Typhlodromus pyri, exposed 7 days to a 51.1 % (w/w) aqueous solution of cyanamide:

Nominal concentrations of cyanamide (g as/ha)	% Mortality (Abbott's correction)	% Reduction in reproduction
100	8.8 %	10.0 %
173	10.5 %	28.0 %
300	36.8 %	36.0 %*
520	38.6 %	16.0 %
900	91.2 %	Not determined
Endpoints
LR50	445.6

Validity criteria fulfilled:

yes

Conclusions:

The LR50 value was determined to be 445.6 g pure cyanamide/ha with 95 % confidence limits of 382.6 and 518.9 g cyanamide/ha.
The reproduction of surviving mites was significantly reduced at 587 g test substance/ha. However at a rate of 1018 g test substance/ha, reproductive performance was not significantly different from the control. Therefore, it was concluded that the result observed in this study was not dose related.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and followed the methodical improvements of standard laboratory tests for determining the side effects of agrochemicals on predatory mites (Louis & Ufer, 1995) and current improvements of the ring-test group (Blümel et al., 1999).

Adult mortality and reproductive effects (expressed as fertile egg production/female) were evaluated following a 1-week exposure of Typhlodromus pyri to dried residues of SKW Cyanamid L500 on treated glass plates. The application rates were 196 g, 339 g, 587 g, 1018 g and 1761 g of a 51.1 % (w/w) aqueous solution of cyanamide per ha and a spray volume of 200 L water/ha (equivalent to 100, 173, 300, 520 and 900 g pure active substance cyanamide. An untreated control and toxic standard (3.6 g Dimethoate/ha) were also tested.

Mortality in the control and toxic standard groups was 5% and 89.5%, respectively.

The LR50 value was determined to be 445.6 g pure cyanamide/ha with 95 % confidence limits of 382.6 and 518.9 g cyanamide/ha.

The reproduction of surviving mites was significantly reduced at 587 g test substance/ha.  However at a rate of 1018 g test substance/ha, reproductive performance was not significantly different from the control.  Therefore, it was concluded that the result observed in this study was not dose related.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 5

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

LR50

Effect conc.:

< 39.14 other: Kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

Adult mortality in the control and toxic standard groups was 0% and 100%, respectively. A 51.1% (w/w) aqueous solution of cyanamide at 39.14 kg product/ha (the maximum recommeded field application rate) resulted in corrected mortality of 100%.
No reproduction testing was performed due 100% mortality in the test item group.

Results with reference substance (positive control):

Adult mortality in the toxic standard group 100%

Reported statistics and error estimates:

Not indicated

The effects on mortality, reproduction, and reduction in beneficial capacity of the aphid parasitoid, Aphidius rhopalosiphi, exposed to a 51.1% (w/w) aqueous solution of cyanamide in the laboratory:

Test substance	51.1% (w/w) aqueous solution of cyanamide
Test organism	Aphidius rhopalosiphi
Exposure	48 h, glass plates
% Mortality (Abbott's correction) (20 kg cyanamide/ha)	100%
% Reduction in Reproduction (20 kg cyanamide/ha)	no reproductive testing was performed
% Reduction in Beneficial Capacity (20 kg cyanamide/ha)	100%

Validity criteria fulfilled:

yes

Conclusions:

100% mortality was observed in the single test concentration (39.14 kg product/ha (equivalent to 20 kg cyanamide/ha).
No reproduction testing was performed due to 100% mortlaity in the test item group.
It was concluded that the test item was harmful to Aphidius rhopalosiphi at 20 kg a.i./ha under the conditions of the test.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and followed the Guideline for testing the effect of pesticides on Aphidius matricariae (Polgar, 1988).

Adult mortality and reproductive effects (parasitism rate) were evaluated following a 48-hour exposure period to dried residues of SKW Cyanamid L500 (a 51.1% (w/w) aqueous solution of cyanamide) on treated glass plates. The application rate was at 39.14 kg of the 51.1% (w/w) aqueous solution of cyanamide per ha in a spray volume of 200 L water/ha. This rate was equivalent to the maximum proposed field rate of 39.14 kg SKW Cyanamid L500 /ha (relevant for the use of cyanamide as herbicide in onions and leek). This equates to 20 kg cyanamide (active ingredient in SKW Cyanamid L500)/ha.

An untreated control and a toxic standard (0.34 g dimethoate/ha) were also tested. Adult mortality in the control and toxic standard groups was 0% and 100%, respectively, confirmed the validity of the test.

A 51.1% (w/w) aqueous solution of cyanamide at 39.14 kg product/ha (the maximum recommeded field application rate) resulted in corrected mortality of 100%.

No reproduction testing was performed due to 100% mortality in the test item group.

It can be concluded that a 51.1% (w/w) aqueous solution of cyanamide applied to glass plates at 39.14 kg product/ha (equivalent to 20 kg cyanamide/ha) was harmful to Aphidius rhopalosiphi under the conditions of the test.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 6

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

21 d

Dose descriptor:

LR50

Effect conc.:

< 39.14 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Pre-imaginal mortality in the control was 16.7%. A 51.1% (w/w) aqueous solution of cyanamide at 39.14 kg product/ha resulted in corrected mortality of 92%.
No reproduction testing was performed due to high mortality in the test substance group.

Results with reference substance (positive control):

Pre-imaginal mortality in the toxic standard group was 84.0 %.

Reported statistics and error estimates:

Not indicated

The effects on pre-imaginal mortality and reproduction of the green lacewing, Chrysoperla carnea, exposed to 51.1% (w/w) aqueous solution of cyanamide in the laboratory:

Test substance	51.1% (w/w) aqueous solution of cyanamide
Test organism	Chrysoperla carnea
Exposure	21d, glass plates
% Mortality (Abbott’s correction) (20 kg pure Cyanamide/ha)	92 %
% Reduction in reproduction (20 kg pure Cyanamide/ha)	not determined

Validity criteria fulfilled:

yes

Conclusions:

A 51.1 % (w/w) aqueous solution of cyanamide applied to glass plates at 39.14 kg /ha (corresponding to 196 g product/L and equivalent to 20 kg pure cyanamide/ha), resulted in corrected mortality of 92 %.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and following a laboratory method for testing side-effects of pesticides on larvae of the green lacewing, Chrysoperla carnea Steph. (Bigler, 1988).

Pre-imaginal mortality and reproductive effects (expressed as fertile egg production/female) were evaluated following a 21-day exposure of larval Chrysoperla carnea to dried residues of SKW Cyanamid L500 (a 51.1% (w/w) aqueous solution of cyanamide) on treated glass plates. The application rate was at 39.14 kg of the 51.1% (w/w) aqueous solution of cyanamide per ha in a spray volume of 200 L water/ha.

An untreated control and a toxic standard (15.9 g Dimethoate/ha) were also tested.

Pre-imaginal mortality in the control and toxic standard groups was 16.7% and 84%, respectively.

A 51.1% (w/w) aqueous solution of cyanamide at 39.14 kg product/ha resulted in corrected mortality of 92%.

No reproduction testing was performed due to high mortality in the test substance group.

It can be concluded that exposure of the lacewing Chrysoperla carnea to dried residues of SKW cyanamide L 500 at 39.14 kg/ha caused significant affects.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 7

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

14 d

Dose descriptor:

LR50

Effect conc.:

< 20 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

Mortality in the control was 8.8% . A 51.1% (w/w) aqueous solution of cyanamide applied at 39.14 kg product/ha (proposed field application rate) resulted in a corrected mortality of 100%.
Due to 100% mortality in the single test concentration effects of the test item on food consumption could not be assessed

Results with reference substance (positive control):

Mortality in the toxic standard group was 94.1%.

Reported statistics and error estimates:

Not indicated

The effects on mortality and food consumption of the wolf spider,Pardosa spec.exposed to a 51.1 % (w/w) aqueous solution of cyanamide in the laboratory:

Test substance	51.1 % (w/w) aqueous solution of cyanamide
Test organism	Pardosa spec.
Exposure	14 d, sand
% Mortality (Abbott’s correction) (20 kg pure cyanamide/ha)	100 %
% Reduction in Food Consumption (20 kg pure cyanamide/ha)	100 %*

* Note: Due to 100% mortality in single test concentration effects on food consumption could not be assessed.

Validity criteria fulfilled:

yes

Conclusions:

A 51.1% (w/w) aqueous solution of cyanamide applied to sand surface at 39.14 kg product/ha resulted in a corrected mortality of 100%. Due to 100% mortality in the single test concentration effects on food consumption could not be assessed.
According to these results, a 51.1% (w/w) aqueous solution of cyanamide can be assessed as harmful to Pardosa spec. when applied at rate of 39.14 kg product/ha, equivalent to 20 kg pure cyanamide/ha.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

Mortality and food consumption effects were evaluated following a 14-day exposure of Pardosa spec. to SKW Cyanamid L500 (a 51.1 % (w/w) aqueous solution of cyanamide). The test substance was applied to the surface of quartz sand with the test organism present at 97.85 g product/L. The application rate was at 39.14 kg of the 51.1 % (w/w) aqueous solution of cyanamide per ha in a spray volume of 500 L water/ha. This equates to 20 kg pure active ingredient cyanamide/ha. An untreated control and a toxic standard (150 ml Thiodan 35 EC/ha) were also tested.

Mortality in the control and toxic standard groups was 8.8% and 94.1%, respectively.

A 51.1% (w/w) aqueous solution of cyanamide applied at 39.14 kg product/ha (proposed field application rate) resulted in a corrected mortality of 100%. Since all spiders in the single test concentration were affected, food consumption could not be assessed.

A 51.1% (w/w) aqueous solution of cyanamide applied to sand surface at 39.14 kg product/ha resulted in a corrected mortality of 100% and reduction in food consumption of 100% relative to the control group.

According to these results, a 51.1% (w/w) aqueous solution of cyanamide can be assessed as harmful to Pardosa spec. when applied at rate of 39.14 kg product/ha, equivalent to 20 kg pure cyanamide/ha.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 8

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

<= 20 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: Mortality and food consumption

Details on results:

Mortality in the control a 0%. A 51.1% (w/w) aqueous solution of cyanamide applied at 39.14 kg product/ha (proposed field application rate) resulted in corrected mortality of 0%. No significant reduction in food consumption (1.4%) as a result of exposure to the test substance was observed.

Results with reference substance (positive control):

Mortality in the toxic standard groups was 93.3%.

Reported statistics and error estimates:

Not indicated

The effects on mortality and food consumption of the ground beetle, Poecilus cupreus exposed to a 51.1 % (w/w) aqueous solution of cyanamide in the laboratory:

Test substance	51.1 % (w/w) aqueous solution of cyanamide
Test organism	Poecilus cupreus
Exposure	14 d, sand
% Mortality (Abbott’s correction) (20 kg pure cyanamide/ha)	0 %
% Reduction in Food Consumption (20 kg pure cyanamide/ha)	1.4 %

Validity criteria fulfilled:

yes

Conclusions:

A 51.1% (w/w) aqueous solution of cyanamide applied to sand surface at 39.14 kg product/ha resulted in a corrected mortality of 0% and reduction in food consumption of 1.4% relative to the control group.
According to these results, a 51.1% (w/w) aqueous solution of cyanamide can be assessed as harmless to Poecilus cupreus when applied at rate of 39.14 kg product/ha, equivalent to 20 kg pure cyanamide/ha.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted in accordance with GLP and following BBA guideline part YVI, 23 -2.1.8 (1991).

Mortality and food consumption effects were evaluated following a 14-day exposure of Poecilus cupreus L. to SKW Cyanamid L500 (a 51.1% (w/w) aqueous solution of cyanamide). The test substance was applied to the surface of quartz sand with the test organism present at 97.85 g product/L. The application rate was at 39.14 kg of the 51.1% (w/w) aqueous solution of cyanamide per ha in a spray volume of 400 L water/ha. This equates to 20 kg pure active ingredient cyanamide/ha. An untreated control and a toxic standard (1 L Afugan 30 EC/ha) were also tested.

Mortality in the control and toxic standard groups was 0% and 93.3%, respectively. A 51.1% (w/w) aqueous solution of cyanamide applied at 39.14 kg product/ha (proposed field application rate) resulted in corrected mortality of 0%. No significant reduction in food consumption (1.4%) as a result of exposure to the test substance was observed.

A 51.1% (w/w) aqueous solution of cyanamide applied to sand surface at 39.14 kg product/ha resulted in a corrected mortality of 0% and reduction in food consumption of 1.4% relative to the control group.

According to these results, a 51.1% (w/w) aqueous solution of cyanamide can be assessed as harmless to Poecilus cupreus when applied at rate of 39.14 kg product/ha, equivalent to 20 kg pure cyanamide/ha.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 9

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

LR50

Effect conc.:

432 g/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

313 g/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

LR100

Effect conc.:

2 500 g/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

Adult mortality in the control and toxic standard groups was 7.5% and 100%, respectively.
The LR50 value was determined to be 432.1 g pure cyanamide/ha with 95 % confidence limits of 365 and 511 g cyanamide/ha.
The reproduction of surviving parasitoids was statistically not significantly affected at rates up to 625 g cyanamide/ha (equivalent to 316 g pure cyanamide/ha)

Results with reference substance (positive control):

Adult mortality in the toxic standard group was 100%.

Reported statistics and error estimates:

95 % confidence limits

Effects on parasitoids (Aphidius rhopalosiphi) exposed for 48 h to a 50.6 % (w/w) aqueous solution of cyanamide:

Nominal concentration of cyanamide (g as/ha)	% Mortality (Abbott's correction)	% Reduction in reproduction
158	-5.4	25.0
316	27.0	13.2
633	81.1	not determined
1265	100.0	not determined
2530	100.0	not determined
Endpoints
LR50	432

Validity criteria fulfilled:

yes

Conclusions:

The LR50 value was determined to be 432.1 g pure cyanamide/ha with 95 % confidence limits of 365 and 511 g cyanamide/ha.
The reproduction of surviving parasitoids was statistically not significantly affected at rates up to 625 g cyanamide product/ha (equivalent to 316.3 g pure cyanamide/ha).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and following the guideline for testing the effect of pesticides on Aphidius matricariae (Polgar, 1988).

Adult mortality and reproductive effects (parasitism rate) in Aphidius rhopalosiphi were evaluated following a 48-hour exposure period to dried residues of SKW Cyanamid L500 on treated glass plates. The application rates were equivalent to 313, 625, 1250, 2500 and 5000 g of a 50.6 % (w/w) aqueous solution of cyanamide per ha and a spray volume of 200 L water/ha. These rates equate to 158, 316, 633, 1265 and 2530 pure active substance cyanamide. An untreated control and toxic standard (0.3 mL /ha) were also tested.

Adult mortality in the control and toxic standard groups was 7.5% and 100%, respectively.

The LR₅₀value was determined to be 432.1 g pure cyanamide/ha with 95 % confidence limits of 365 and 511 g cyanamide/ha. The reproduction of surviving parasitoids was statistically not significantly affected at rates up to 625 g cyanamide/ha (equivalent to 316.3 g pure cyanamide/ha).

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 10

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

28 d

Dose descriptor:

other: ER50

Effect conc.:

0.93 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: Mean no. hatched beetles of F1 generation per introduced female

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

<= 0.3 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Key result

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

<= 0.59 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Details on results:

The result of the control group indicated that the test organisms were in a good condition (average number of hatched beetles of the F1-generation per replicate: 845 (validity criteria: 400 hatched beetles per replicate)).
Statistical analysis of reproduction parameters revealed significant differences concerning the reproductive capacity between the control and all test item treatment groups. The NOEC was considered to be <=0.3 kg/ha (lowest tested concentration), equivalent to 0.59 kg product/ha.
The ER50 was calculated to be 0.93 kg a.i./ha with 95% confidence limits ranging from 0.56 to 1.49 kg a.i./ha, corresponding to 1.82 kg product/ha with confidence limits ranging from 1.10 to 2.92 kg product/ha.

Results with reference substance (positive control):

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (average number of hatched beetles of the F1-generation per replicate: 11 corresponding to 98.7% reduction of reproductive capacity).

Reported statistics and error estimates:

The test groups were statistically significantly different from the control (FISHER’S Exact Binomial-test, 1-sided, p≤0.05)

Effects of Cyanamide 51.1 % (w/w) aqueous solution on reproduction of Aleochara bilineata exposed over 28 days in the laboratory:

Application rate [kg a.i./ha]	Reproductive capacity
	mean number of hatched beetles of the F1-generation per replicate	mean number of hatched beetles of the F1-generation per introduced female (10)	mean number of hatched beetles/ host pupa	Parasiti-sation rate P (%)	Reduction of reproductive capacity (relative to control) R (%)
Control	845	84.5	0.563	56.3	-
0.3 0.9 2.5 7.3 21.2	691* 495* 70* 9* 0.5*	69.1 49.5 7.0 0.9 0.05	0.461 0.330 0.047 0.006 0.0003	46.1 33.0 4.7 0.6 0.03	18.2 41.4 91.7 98.9 99.9
Reference item	11	1.1	0.007	1.1	98.7

Number of hatched beetles was determined over a period of 70 days.         

^*Statistically significantly different from the control (Student-t-test for homogeneous variances withBonferroniadjustment, one-sided, p < 0.05)

Validity criteria fulfilled:

yes

Conclusions:

The ER50 was calculated to be 0.93 kg a.i./ha with 95% confidence limits ranging from 0.56 to 1.49 kg a.i./ha, corresponding to 1.82 kg product/ha with confidence limits ranging from 1.10 to 2.92 kg product/ha.
Statistical analysis of reproduction parameters showed significant differences between the control and all test item treatment groups. Therefore, the NOEC was considered to be <=0.59 kg prodcut/ha.

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted following the IOBC guideline (Grimm et al, 2000).

The test item Cyanamid L500 was tested under laboratory conditions via exposure of adults of the rove beetle Aleochara bilineata GYLL. over a period of 28 days to spray residues with rates of 0.3, 0.9, 2.5, 7.3 and 21.2 kg a.i./ha (corresponding to 0.59, 1.76, 4.89, 14.29 and 41.49 kg product/ha) in 400 L deionised water/ha applied on quartz sand. The control was treated with deionised water (400 L/ha). Dimethoate EC 400 (1.5 L product/ha, corresponding to 600 g a.i./ha, in 400 L/ha of water) was used as a toxic reference treatment.

Adults of Aleochara bilineata GYLL. were exposed in two replicates, each replicate consisting of 10 pairs of beetles (10 females and 10 males) per treatment group to the spray residues of the test item, reference item and control treatments, respectively. During the assessments, the beetles were fed with deep frozen larvae of Chironomus spp. As hosts onion flies were added to the test containers. The number of hatched beetles of the F1 generation was recorded over a period of 70 days. From these data the endpoint reproductive capacity was calculated.

The result of the control group indicated that the test organisms were in a good condition (average number of hatched beetles of the F1-generation per replicate: 845 (validity criteria: 400 hatched beetles per replicate)). The results of the toxic standard group indicated that the test system was sensitive to harmful substances (average number of hatched beetles of the F1-generation per replicate: 11 corresponding to 98.7% reduction of reproductive capacity).

Statistical analysis of reproduction parameters revealed significant differences concerning the reproductive capacity between the control and all test item treatment groups. Therefore, the NOEC was considered to be <=0.3 kg ai/ha (lowest tested concentration), equivalent to 0.59 kg prodcut/ha.

The ER50 was calculated to be 0.93 kg a.i./ha with 95% confidence limits ranging from 0.56 to 1.49 kg a.i./ha, corresponding to 1.82 kg product/ha with confidence limits ranging from 1.10 to 2.92 kg product/ha.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 11

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

18 d

Dose descriptor:

LR50

Effect conc.:

3.4 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Key result

Duration:

18 d

Dose descriptor:

LR50

Effect conc.:

6.65 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

2.5 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Key result

Duration:

18 d

Dose descriptor:

NOEC

Effect conc.:

4.89 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Details on results:

The result of the control group indicated that the test organisms were in a good condition (mortality: 0%)

Statistically significant differences in mortality after 18 days were observed in the 7.3 and 21.2 kg a.i./ha test item treatment groups compared to the control group.

The LR50 was calculated to be 3.4 kg a.i./ha with 95 % confidence limits ranging from 2.0 to 5.9 kg a.i./ha, corresponding to 6.65 kg product/ha with confidence limits ranging from 3.91 to 11.55 kg product/ha.

Results with reference substance (positive control):

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 65.0%). Hence, the study is considered valid.

Reported statistics and error estimates:

The test groups were statistically significantly different from the control (FISHER’S Exact Binomial-test, 1-sided, p≤0.05)

Effects of cyanamide 51.1 % (w/w) aqueous solution on mortality of Chrysoperla carnea exposed over 18 days in the laboratory:

Application rate [kg a.i./ha]	Mortality after 18 days [%]
Control	0
0.3	0
0.9	0
2.5	25.0
7.3	100^*
21.2	100^*
Reference item	65.0

^*Statistically significantly different from the control (Fisher’s Exact Binomial Test, 1-sided, p < 0.05)

Validity criteria fulfilled:

yes

Conclusions:

The LR50 of Cyanamid L500 to Chrysoperla carnea STEPH. was calculated to be 3.4 kg a.i./ha with 95 % confidence limits ranging from 2.0 to 5.9 kg a.i./ha, (corresponding to 6.65 kg product/ha with confidence limits ranging from 3.91 to 11.55 kg product/ha).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted following the IOBC guideline (Vogt et al, 2000).

The test item Cyanamid L500 was tested under laboratory conditions via exposure of larvae of the green lacewing Chrysoperla carnea (STEPH.) to spray residues with rates of 0.3, 0.9, 2.5, 7.3 and 21.2 kg a.i./ha (corresponding to 0.59, 1.76, 4.89, 14.29 and 41.49 kg product/ha) in 200 L deionised water/ha applied on glass plates. The control was treated with deionised water (200 L/ha). Dimethoate EC 400 (40 mL product/ha in 200 L water/ha) was used as a toxic reference treatment.

The larvae of C. carnea were exposed in 20 replicates of one larva (per treatment group) to freshly applied dried residues of the test item, reference item and control treatments, respectively. During the assessments the larvae were fed with UV-sterilized eggs of Sitotroga cerealella. The number of surviving larvae and hatched adults was recorded over a period of 18 days. From these data the endpoint mortality was calculated.

The result of the control group indicated that the test organisms were in a good condition (mortality: 0%). The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 65.0%). Hence, the study is considered valid.

Statistically significant differences in mortality after 18 days were observed in the 7.3 and 21.2 kg a.i./ha test item treatment groups compared to the control group. Therefore, the NOEC (mortality) was considered to be 2.5 kg ai/ha (equivalent to 4.89 kg product/ha)

The LR50 was calculated to be 3.4 kg a.i./ha with 95% confidence limits ranging from 2.0 to 5.9 kg a.i./ha, corresponding to 6.65 kg product/ha with confidence limits ranging from 3.91 to 11.55 kg product/ha.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 12

Endpoint:

toxicity to terrestrial arthropods: short-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

14 d

Dose descriptor:

LR50

Effect conc.:

2.1 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

0.9 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

2.5 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

food consumption

Remarks on result:

other: Mean no. consumed flies/spider/day

Details on results:

The results of the control group indicated that the test organisms were in a good condition (mortality: 0 %, food uptake: 3.3 flies/spider/day).

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 75.0 %). Hence, the test was considered valid.

Statistical analysis revealed significant differences concerning the mortality after 14 days at 2.5, 7.3 and 21.2 kg a.i./ha compared to the control.

The LR50 was calculated to be 2.1 kg a.i./ha with 95 % confidence limits ranging from 1.7 to 2.7 kg a.i./ha, corresponding to 4.12 kg product/ha with confidence limits ranging from 3.33 to 5.28 kg product/ha.

Regarding the feeding rate in the test item treatment groups, there was a statistically significant difference between the 2.5, 7.3 and 21.2 kg a.i./ha treatment groups and the control group in the 1st week and between the 7.3 and 21.2 kg a.i./ha treatment group in the total test period of 14 days. There was no statistically significant difference in the 2nd week between the treatment groups 0.3, 0.9 and 2.5 kg a.i./ha and the control group.

No abnormal behaviour and no damages of the spiders were observed in all test item treatment groups compared to the control group.

Results with reference substance (positive control):

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 75.0 %). Hence, the test was considered valid.

Reported statistics and error estimates:

The test groups were statistically significantly different from the control (FISHER’S Exact Binomial-test, 1-sided, p≤0.05)

Effects of cyanamide 51.1 % (w/w) aqueous solution on mortality and feeding capacity of Pardosa spec. exposed 14 days in the laboratory:

Application rate [kg a.i./ha]	Mortality after 14 days [%]	Feeding capacity (Food uptake)
		Mean number of consumed flies/spider/day	Relative to control [%]	Reduction relative to control [%]
Control	0	3.3	-	-
0.3	0	3.1	93.9	6.1
0.9	5.0	3.0	90.9	9.1
2.5	60.0^*	2.3	69.7	30.3
7.3	100^*	0.6^*	18.2	81.8
21.2	100^*	0.1^*	3.0	97.0
Reference item	75.0	1.6	48.5	51.5

Validity criteria fulfilled:

yes

Conclusions:

The LR50 (median lethal rate) of Cyanamid L500 to Pardosa spp. was calculated to be 2.1 kg a.i./ha with 95 % confidence limits ranging from 1.7 to 2.7 kg a.i./ha, corresponding to 4.12 kg product/ha with confidence limits ranging from 3.33 to 5.28 kg product/ha. The NOEC (mortality) was considered to be 0.9 kg ai/ha (equivalent to 1.76 kg product/ha in 400 L water/ha).
The NOEC (feeding rate) was considered to be 2.5 kg ai/ha (equivalent to 4.89 kg product/ha in 400 L water/ha).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted following IOBC (Heimbach et al, 2000) guideline modified according to screening test design.

The test item Cyanamid L 500 was tested under laboratory conditions via exposure of adults of the spider Pardosa spp. to spray residues with rates of 0.3 – 0.9 – 2.5 – 7.3 and 21.2 kg a.i./ha (corresponding to 0.59 – 1.76 – 4.89 – 14.29 and 41.49 kg product/ha) in 400 L deionised water/ha applied on quartz sand. The control was treated with deionised water (400 L/ha). Dimethoate EC 400 (1.5 L product/ha, corresponding to 600 g a.i./ha, in 400 L/ha of water) was used as a toxic reference treatment.

Adults of the spider Pardosa spp. were exposed in 20 replicates of one animal each (10 females and 10 males) per treatment group to the spray residues of the test item, reference item and control treatments, respectively. During the assessments the spiders were fed with frozen adult Drosophila spp. The number of dead and living spiders, the number of consumed flies per evaluation period, as well as behavioural impacts were recorded over a period of 14 days. From these data the endpoints mortality and effect on food uptake were calculated.

The results of the control group indicated that the test organisms were in a good condition (mortality: 0 %, food uptake: 3.3 flies/spider/day).

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 75.0 %). Hence, the test was considered valid.

Statistical analysis revealed significant differences concerning the mortality after 14 days at 2.5, 7.3 and 21.2 kg a.i./ha compared to the control.

The LR50 was calculated to be 2.1 kg a.i./ha with 95 % confidence limits ranging from 1.7 to 2.7 kg a.i./ha (equivalent to 4.12 kg product/ha with confidence limits ranging from 3.33 to 5.28 kg product/ha).

Regarding the feeding rate in the test item treatment groups, there was a statistically significant difference between the 2.5, 7.3 and 21.2 kg a.i./ha treatment groups and the control group in the 1st week and between the 7.3 and 21.2 kg a.i./ha treatment group in the total test period of 14 days. There was no statistically significant difference in the 2nd week between the treatment groups 0.3, 0.9 and 2.5 kg a.i./ha and the control group.

No abnormal behaviour and no damages of the spiders were observed in all test item treatment groups compared to the control group.

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 13

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

48 h

Dose descriptor:

LR50

Effect conc.:

0.62 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Key result

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

0.34 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 0.58 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Remarks on result:

other: mean no. mummies/female

Details on results:

- The result of the control group indicated that the test organisms were in a good condition (mortality: 3.3%, reproduction: 13.6 mummies/female)

- Statistically significant differences in mortality after 48 hours were observed in the 0.58, 0.98 and 1.67 kg ai/ha test item treatment groups compared to the control group. The LR50 was calculated to be 0.62 kg ai/ha with 95% confidence limits ranging from 0.54 to 0.72 kg ai/ha. The NOEC was determined to be 0.34 kg ai/ha (equivalent to 0.67 kg product/ha in 400 L water/ha)

- The behaviour assessments showed statistically significant differences regarding the number of wasps sitting on the treated plants in all test item groups compared to the control group within 3 hours after start of the exposure. In the treatment groups more wasps were found on the sand and/or on the walls of the cylinder

- There were no statistically significant effects of Cyanamid L500 on reproduction (mean number of mummies/female) at all test item treatment groups which were tested compared to the control. Therefore, the NOEC was considered to be >/=0.58 kg ai/ha (equivalent to 1.14 kg product/ha in 400 L water/ha).

Results with reference substance (positive control):

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 100%). Hence, the study is considered valid.

Reported statistics and error estimates:

A statistically significantly difference between the tets groups and the control groups was examined by the FISHER’S Exact Binomial Test, 1-sided, p= 0.05

Effects of cyanamide 51.1% (w/w) aqueous solution on mortality of Aphidius rhopalosiphiexposed over 14 days in the laboratory:

Treatment [kg ai/ha]	Mortality after 48 hrs [%]^1	Reproduction
		Mean number of mummies/female	Relative to control [%]	Reduction relative to control [%]
Control	3.3	13.6	-	-
0.2	0	14.1	103.7	0 (+ 3.7)
0.34	17.2	14.1	103.7	0 (+ 3.7)
0.58	48.3 *	12.3	90.4	9.6
0.98	75.9*	n.d	-	-
1.67	96.6*	n.d	-	-
Dimethoate (10 mL product/ha)	100	-	-	-

Validity criteria fulfilled:

yes

Conclusions:

The LR50 (48 h) of Cyanamid L500 to Aphidius rhopalosiphi (DESTEFANI-PEREZ) was calculated to be 0.62 kg ai cyanamide/ha. The NOEC (mortality) was determined to be 0.34 kg ai/ha (equivalent to 0.67 kg product/ha in 400 L water/ha).
No significant effect on reproduction was observed up to and including the highest tested concentrations of 0.58 kg ai/ha (equivalent to 1.14 kg product/ha in 400 L water/ha).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and followed the IOBC draft guideline (Mead-Briggs et al, 2006).

The test item was tested under extended laboratory conditions via exposure of the adults of the parasitic wasp Aphidius rhopalosiphi (DESTEFANI-PEREZ) to spray residues on potted barley plants. The test item was applied at rates of 0.2, 0.34, 0.58, 0.98 and 1.67 kg ai/ha (corresponding to 0.39, 0.67, 1.14, 1.92 and 3.27 kg product/ha) in 400 L water/ha. The control was treated with deionised water (400 L/ha) in the same way as the test item treatment. Dimethoate EC 400 was used as toxic reference treatment.

Adults were exposed in 6 replicates of 5 female wasps (per treatment group) to the residues of the test item, reference item (only 1 replicate) and control treatments, respectively. During the mortality test the wasps were fed with aqueous fructose solution (10% w/w). Aphids (Rhopalosiphum padi) were used as host organisms. The number of surviving wasps, behaviour and position and number of parasited aphids (mummies) were recorded over a period of 14 days. Mortality and fecundity were calculated as endpoints.

The result of the control group indicated that the test organisms were in a good condition (mortality: 3.3%, reproduction: 13.6 mummies/female). The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 100%). Hence, the study is considered valid.

Statistically significant differences in mortality after 48 hours were observed in the 0.58, 0.98 and 1.67 kg ai/ha test item treatment groups compared to the control group, therefore the NOEC (mortality) was considered to be 0.34 ka ai/ha (equivalent to 0.67 kg prodcut/ha in 400 L water/ha). The LR50 was calculated to be 0.62 kg ai/ha with 95% confidence limits ranging from 0.54 to 0.72 kg ai/ha.

The behaviour assessments showed statistically significant differences regarding the number of wasps sitting on the treated plants in all test item groups compared to the control group within 3 hours after start of the exposure. In the treatment groups more wasps were found on the sand and/or on the walls of the cylinder.

There were no statistically significant effects of Cyanamid L500 on reproduction (mean number of mummies/female) at all test item treatment groups which were tested compared to the control. Therefore, the NOEC (reproduction) was considered to be >/=0.58 kg/ai/ha (equivalent to 1.14 kg product/ha in 400 L water/ha).

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 14

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

7 d

Dose descriptor:

LR50

Effect conc.:

2.07 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

1.02 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

7 d

Dose descriptor:

LR50

Effect conc.:

> 1.73 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Duration:

7 d

Dose descriptor:

NOEC

Effect conc.:

1.02 other: kg/ha

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Details on results:

- The results of the control group indicated that the test organisms were in a good condition
- All validity criteria according to BLÜMEL et al. (2000) for conducting the laboratory test with Typhlodromus pyri and adapted to the extended laboratory test were met
- Statistical analysis (FISHER`S Exact Binomial-test, 1-sided, p≤ 0.05) revealed significant differences concerning the mortality after 7 days between the control and the 1.17, 2.00 and 3.39 kg prodcut/ha in 200 L water/ha (equivalent to 1.73, 2.95 and 5.0 kg ai/ha) test item treatment groups. The LR50 (median lethal rate) of Cyanamid L500 to Typhlodromus pyri was calculated to be 2.07 kg ai/ha, with 95 % confidence limits ranging from 1.92 to 2.23 kg ai/ha.

Results with reference substance (positive control):

The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 60.6 %).

Reported statistics and error estimates:

A statistically significantly difference between the tets groups and the control groups was examined by the FISHER’S Exact Binomial Test, 1-sided, p≤ 0.05

Effects of Cyanamid L500 on reproduction of Typhlodromus pyrion bean leaf discs:

Test group (kg test item/ha)	Mean number of eggs/female	Relative to control [%]	Reduction relative to control [%]
Control	7.22
0.6	6.77	93.8	6.2
1.02	6.89	95.4	4.6
1.73	6.28^*	87.0	13.0
2.95	n.a.	-	-
5.0	n.a.	-	-
Toxic standard	-	-	-

^*statistically significantly different from the control (p 0.05)

CL confidence limit

n.a. not assessed because of > 50 % corrected mortality 

Validity criteria fulfilled:

yes

Conclusions:

In a dose-response extended laboratory study with the predatory miite, Typhlodromus pyri, the LR50 was determined to be 2.07 kg ai/ha. The NOEC (mortality) was determined to be 1.02 kg ai/ha (equivalent to 2.0 kg product/ha in 200 L/ha water).
The effects on reproduction were <50 % up to and including 1.73 kg ai/ha, the highest rate tested for effects on reproduction, therefore, the LR50 was considered to be >1.73 kg ai/ha (equivalent to 3.39 kg product/ha in 200 L/ha water). The NOEC (reproduction) was determined to be 1.02 kg ai/ha (equivalent to 2.0 kg product/ha in 200 L/ha water).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The study was conducted according to GLP and followed the IOBC guideline (Blumel et el, 2000).

Protonymphs of T. pyri were exposed in 5 replicates of 20 mites (per treatment group) to the spray residues of the test item, reference item (Dimethoate EC 400, 15 mL) and control treatments, applied on bean leaf discs (Phaseolus vulgaris). The test item was applied at rates of 0.6, 1.02, 1.73, 2.95 and 5.0 kg ai/ha (corresponding to 1.17, 2.00, 3.39, 5.77 and 9.78 kg product/ha in 200 l water/ha0. During the assessments the predatory mites were fed with pollen (Pinus nigra and Betula pendula). The number of surviving, dead and escaped predatory mites and the number of eggs laid per viable female per evaluation period as well as behavioural impacts were recorded over a period of 14 days.

On day 3, 7, 9, 11 and 14 after the application, the number of surviving predatory mites was counted (from 7th day onward differentiated according to the sex), dead mites were recorded and removed. The number of laid eggs was determined on days 9, 11 and 14. Any eggs found on day 7 were removed and not counted in the fecundity assessment. From these data the endpoints mortality and effect on reproduction were calculated. The dose-response relationship regarding mortality (LR50) was determined.

All validity criteria according to BLÜMEL et al. (2000) for conducting the laboratory test with Typhlodromus pyri and adapted to the extended laboratory test were met. The results of the control group indicated that the test organisms were in a good condition (mortality: 1%, reproduction: 7.22 eggs/female). The results of the toxic standard group indicated that the test system was sensitive to harmful substances (corrected mortality: 60.6%).

Statistical analysis revealed significant differences concerning the mortality after 7 days between the control and the 1.73, 2.95 and 5.0 kg ai/ha test item treatment groups. The LR50 (median lethal rate) of Cyanamid L500 to Typhlodromus pyri was calculated to be 2.07 kg ai/ha, with 95% confidence limits ranging from 1.92 to 2.23 kg ai/ha. The NOEC (mortality) was determined to be 1.02 ka ai/ha (equivalent to 2.0 kg product/ha in 200 L/ha water).

The effects on reproduction were below 50 % up to and including 1.73 kg ai/ha, the highest rate tested for effects on reproduction, therefore, the LR50 was considered to be >1.73 kg ai/ha (equivalent to 3.39 kg product/ha in 20L/ha water). The NOEC (reproduction) was determined to be 1.02 mh ai/ha (equivalent to 2.0 kg product/ha in 200 L/ha water).

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 15

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

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

Adequacy of study:

supporting study

Justification for type of information:

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.
For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reason / purpose for cross-reference:

read-across source

Duration:

28 d

Dose descriptor:

LC50

Effect conc.:

71.9 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Duration:

28 d

Dose descriptor:

EC50

Effect conc.:

61.7 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

reproduction

Details on results:

Adult mortality:
8% mortality was observed at the control and 4% mortality at the concentration of 0.40 mg a.s./kg soil (dw). At 60.00 and 100.00 mg a.s./kg soil (dw) a mortality of 36 and 84% respectively was determined.
Statistical analysis (Fisher’s Exact test; 1-sided, p ≤ 0.05) revealed a significant difference concerning the mortality between the control and the concentrations of 60.00 and 100.00 mg a.s./kg soil (dw)).
The LC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression to be 71.9 mg a.s./kg soil (dw) (95% confidence limits: 64.2 – 80.4 mg a.s./kg).

Reproduction
Statistical analysis (Williams’ test; 1-sided, p ≤ 0.05) showed a significant difference concerning the number of juveniles between the control and the concentrations of 60.00 and 100.00 mg a.s./kg soil (dw).
The EC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression to be 61.7 mg a.s./kg soil (dw) (95% confidence limits: 61.4 – 62.1 mg a.s./kg).

Reported statistics and error estimates:

- Mortality: mean number of dead or missing adult collembolans as absolute number and as a percentage of the initial number at the start of the test for each concentration and for the control.
- Reproduction: mean number of juvenile collembolans 28 days after application for each concentration and for the control.
- Morphology and behaviour: any pathological or other symptoms or distinct changes in behaviour of the test organisms were reported.

Mortality: The LC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression. The NOECMortality was determined by pair-wise comparisons between treatment and control by the Fisher’s Exact test (p ≤ 0.05, one-sided).

Reproduction: The EC50 value for reproduction was calculated by Probit analysis using Linear Max. Likelihood Regression. For determination of the NOECReproduction data were checked for normality by R/s test procedure 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.

The statistical software package ToxRat Professional Version 2.09 was used for these calculations.

Validity criteria fulfilled:

yes

Remarks:

Av. mortality of the adult test animals in the control: 8% (required: ≤ 20%) Av. reproduction rate per vessel in the control: 586.0 ± 111.1 (required: ≥ 100 juveniles per vessel) Coef. of variation of reproduction in the control: 19.0% (required: ≤ 30%)

Conclusions:

The LC50 was calculated as 71.9 mg a.s./kg soil (dw) (95% confidence limits: 64.2 – 80.4 mg a.s./kg).
The EC50 (reproduction) was calculated as 61.7 mg a.s./kg soil (dw) (95% confidence limits: 61.4 – 62.1 mg a.s./kg).

Justification for read-across for ecotoxicological endpoints:
Environmental fate and exposure are critical factors when assessing environmental risk. Calcium cyanamide is formulated in a slowly dissolving granule (PERLKA) that is applied to agricultural fields as a fertiliser. When dissolved, calcium cyanamide is rapidly converted to (hydrogen) cyanamide. These two substances are similar both in chemical structure and regarding fate in the environment. Cyanamide is the environmentally relevant transformation product upon application of calcium cyanamide (PERLKA) to soil as a fertiliser. Therefore, terrestrial data on cyanamide are considered in the hazard assessment of calcium cyanamide.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Executive summary:

The effects of Cyanamide on survival and reproduction ofFolsomia candidawere tested in natural soil. LUFA St. 2.2 was used as a substrate to investigate possible influences of a natural soil on the bioavailability of Cyanamid L 500 (ca. 50% aqueous solution) which may result in altered effects of the test item on Folsomia candida.

8% mortality was observed at the control and 4% mortality at the concentration of 0.40 mg a.s./kg soil (dw). At 60.00 and 100.00 mg a.s./kg soil (dw) a mortality of 36 and 84%, respectively, was determined.
Statistical analysis (Fisher’s Exact test; 1-sided, p ≤ 0.05) revealed a significant difference concerning the mortality between the control and the concentrations of 60.00 and 100.00 mg a.s./kg soil (dw)).

The LC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression to be 71.9 mg a.s./kg soil (dw) (95% confidence limits: 64.2 – 80.4 mg a.s./kg).
Statistical analysis (Williams’ test; 1-sided, p ≤ 0.05) showed a significant difference concerning the number of juveniles between the control and the concentrations of 60.00 and 100.00 mg a.s./kg soil (dw).
The EC50 value was calculated by Probit analysis using Linear Max. Likelihood Regression to be 61.7 mg a.s./kg soil (dw) (95% confidence limits: 61.4 – 62.1 mg a.s./kg).

This information is used in a read-across approach in the assessment of the target substance.

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Reference 16

Endpoint:

toxicity to terrestrial arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2018-09-21 to Q1/2020 (study ongoing at the date of dossier submission)

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: ISO 23611-2

Version / remarks:

ISO 23611-2 / 2007 / Soil quality – sampling of soil invertebrates – Part 2: Sampling and extraction of micro-arthropods (Collembola and Acarina).

GLP compliance:

yes (incl. QA statement)

Application method:

other: The application of the test item and the control fertiliser was done with a fertiliser spreader for granules

Specific details on test material used for the study:

1. Application: Perlka®; batch-no.: SWSE-18-068; content of a.i.: 45.0 % calcium cyanamide (analysed)
2. Application: Perlka®; batch-no.: SWSE-19-017; content of a.i.: 44.4 % calcium cyanamide (analysed)

Analytical monitoring:

yes

Vehicle:

no

Details on preparation and application of test substrate:

- Method of test material application:
Two applications:
Test item: 1.) 200 kg Perlka®/ha
2.) 400 kg Perlka®/ha
Perlka® at both rates was applied twice, with an interval of 6 months.
Reference item: One application (at the same date as the first application of the test item was performed) of 1.449 L product/ha (corresponding to 0.72 kg a.i. Chlorpyrifos/ha) with 400 L water/ha
Untreated Control: Plots were left untreated
Fertiliser Control: Two applications of Piagran®46 (SKW Piesteritz, total nitrogen content approx. 46.5 % (nominal)) at the same total nitrogen rate as provided by the higher dosed test item (400 kg Perlka®/ha), corresponding to 172.9 kg/ha Piagran

1. Application Date: September 28, 2018
2. Application Date: April 02, 2019
>further applications planned<

Test Conditions: Natural field conditions, soil type silt loam (USDA), 2.56 – 2.64 % total organic carbon, < 0.1 % lime content; pH 5.34 – 5.50; water holding capacity 58.0 – 57.5 %, nitrogen 0.28 – 0.29 %N

Reference Item: Agriclor, 480 g chlorpyrifos/L (nominal)

The application of the test item and the control fertiliser was done with a fertiliser spreader for granules (Hege 80 Parzellenstreuer, System Weihenstephan, Model 422437 with a working width of 1.50 m and 10 gape pipes, Hans-Ulrich Hege Saatzuchtmaschinen, Waldenburg, Germany). Calibration of the fertiliser distributer:
The fertiliser distributer distributed the given amount of fertiliser (i.e. 600 g for an application amount of 400 kg/ha for a working width of 1.5 m) on a given distance i.e. 10 m. The fertiliser spreader was calibrated before application in order to ensure an equal distribution of the granules. The calibration was conducted by measuring the weight of granules over the given distance (10 m) for each of the gate pipes. For the first application this procedure was repeated 3 times with an amount of 300 g and 600 g Perlka®, respectively. The distribution between the 10 gate pipes should be in the range of +/- 20 %.

Furthermore, the variability of the distribution was determined for the test item Perlka® and the fertiliser control Piagran®. Trays (50 x 37.5 cm) were placed on a 10 m section in a distance of 3 m, 5 m and 7 m from the plot margin. Then the fertiliser distributer was filled with a certain amount of Perlka® or Piagran® and the 10 m distance was run. For each substance this was repeated 3 times. Weight in the trays for each run was determined. According to DIN EN 13739-2 the trays have to be passed 2 times for calculating the quality of the distribution the results of two runs (1+2, 2+3, 1+3) were summed up, respectively, for calculating the total coefficient of variability.

Test organisms (species):

other: several species of collembola -> naturally occurring collembola species on a grassland habitat (hay meadow).

Animal group:

Collembola (soil-dwelling springtail)

Details on test organisms:

Naturally occurring collembola species on a grassland habitat (hay meadow).

Study type:

field study

Remarks:

off-crop field study

Limit test:

no

Total exposure duration:

185 d

Remarks:

1. Application Date: September 28, 2018; 2. Application Date: April 02, 2019

Post exposure observation period:

See details on test condition

Test temperature:

See details on test conditions

pH (if soil or dung study):

Sample 1 (western half): 5.34
Sample 2 (eastern half): 5.50

Humidity:

Water Holding Capacity max [g/100 g]:
Sample 1 (western half): 58.0
Sample 2 (eastern half): 57.5

Details on test conditions:

TEST FIELD
The in-life phase of the study was conducted at the test site of Mesocosm GmbH located in Homberg (Ohm), Germany, approximately 80 km direction NNE from Frankfurt/Main. The test field is hold by the FNU Forschungszentrum Neu-Ulrichstein GmbH & Co KG, 35315 Homberg (Ohm), Hessia, Germany. The test field was a hay meadow of ca. 6 hectare in the area Appenrod, district of Gießen, field name “Windelhecke”. Since 2010, the test field has been used as hay meadow without the use of fertilisers and pesticides.
The test site is surrounded by hedges and meadows which are protecting the experimental site from contamination by pesticides which might be used in the surrounding fields.
Field Management was non-GLP, but according to Good Agricultural Practice. During the study no additional pesticide or fertiliser treatment, apart from that of the study, was performed.
For preparation of the test field the grass was mulched on August 27, 2018. Before application, the grass was mown as short as possible on all plots on September 20, 2018. The cuttings were removed to minimize interception of the test item.
Due to a summer characterized by high temperatures and prolonged drought irrigation was performed before the start of the study to ensure suitable sampling conditions. With each irrigation 11 mm/m² water was applied on the following dates August 30, August 31, September 03, September 04 and September 13, 2018 (in total 55 mm/m²).

-Test Design:
The test field was divided in plots with a size of 120 m² (10 x 12 m). Distance between the plots was 3 m for the north-south direction and 5 m for the east-west direction and at least 10 m distance to the fields next to the test site. In total, 20 plots were arranged in a randomized block design

Characterisation of the field:
For characterisation of the field the soil particle size distribution, organic matter content of the soil, soil pH, total N-content and water holding capacity of the A-horizon was determined. The samples were taken once on September 18, 2018 in every quadrant of the test field. With a Pürkhauer-sampler four soil cores from 0-30 cm depth were taken respectively on five places in the east and west half of the field. The 20 soil cores from each half of the field were pooled to one sample resulting in two pooled samples of the field (east half and west half). The soil characterisation was carried out as a GLP-analysis by LUFA Speyer (Landwirtschaftliche Untersuchungs- und Forschungsanstalt Speyer, 67346 Speyer, Germany).

Test Conditions: Natural field conditions, soil type silt loam (USDA), 2.56 – 2.64 % total organic carbon, < 0.1 % lime content; pH 5.34 – 5.50; water holding capacity 58.0 – 57.5 %, nitrogen 0.28 – 0.29 % N.
The particle size distribution according to USDA showed that the soil type can be characterised as silt loam (25.4 – 26.6 % clay, 67.2 – 68.5 % silt, 6.1 – 6.2 % sand).


Climatic conditions
For characterisation of the field the soil particle size distribution, organic matter content of the soil, soil pH, total N-content and water holding capacity of the A-horizon was determined. The samples were taken once on September 18, 2018 in every quadrant of the test field. With a Pürkhauer-sampler four soil cores from 0-30 cm depth were taken respectively on five places in the east and west half of the field. The 20 soil cores from each half of the field were pooled to one sample resulting in two pooled samples of the field (east half and west half). The soil characterisation was carried out as a GLP-analysis by LUFA Speyer (Landwirtschaftliche Untersuchungs- und Forschungsanstalt Speyer, 67346 Speyer, Germany).
Climatic conditions during the first application of the test item, control fertiliser and reference item:
Treatment air temperature [°C] soil temperature [°C] wind speed [m/s] Air humidity [%]
200kg Perlka®/ha min 11.1 11.1 0.2 75.2
time 10:40-11:53 max 13.1 11.5 1.3 81.9
400 kg Perlka®/ha) min 11.8 11.5 0.7 81.2
time 12:02-13:04 max 13.1 11.7 2.6 88.3
Control Fertiliser min 10.8 10.8 0.2 65.6
time 09:25 - 10:26 max 15.2 11.0 1.4 82.4
Reference item min 11.2 11.1 0.2 75.2
time 10:55 - 11:35 max 13.0 11.4 0.8 81.9

Climatic Conditions during the Sampling Dates
A summary of the climatic conditions during the sampling of the specimens in 2018 is given in Table 11. The complete daily mean values of the parameters of the weather station Neu-Ulrichstein for 2018 are presented in the raw data.

Mean values of climatic parameters during first application and the sampling dates in 2018.
Date Days of study average daily air temperature [°C] average daily soil temperature [°C] sum of daily precipitation [mm] average daily air humidity [%]
21/09/2018 -7 15.0 17.7 4.3 73.0
22/09/2018 -6 10.5 13.7 4.6 75.6
23/09/2018 -5 9.7 12.0 36.0 94.6
24/09/2018 -4 8.0 9.9 1.2 76.8
25/09/2018 -3 7.9 11.0 0.0 71.6
26/09/2018 -2 9.5 11.8 0.0 58.8
28/09/2018 0 9.4 10.9 0.0 73.7
01/10/2018 3 7.5 10.2 0.9 73.6
02/10/2018 4 5.7 7.6 2.6 94.6
03/10/2018 5 10.5 10.8 0.0 73.5
04/10/2018 6 9.7 10.0 0.0 80.9
05/10/2018 7 12.6 12.4 0.0 70.5
08/10/2018 10 10.8 13.4 0.0 80.7
09/10/2018 11 13.1 15.4 0.0 84.7
10/10/2018 12 13.6 14.8 0.0 81.6
11/10/2018 13 14.9 15.4 0.0 69.3
12/10/2018 14 17.2 16.8 0.0 68.6
22/10/2018 24 6.7 9.6 0.5 90.7
23/10/2018 25 6.6 8.2 0.0 89.7
24/10/2018 26 11.3 11.1 0.5 89.3
25/10/2018 27 10.3 10.8 0.0 82.5
26/10/2018 28 7.8 9.4 0.2 82.2


Equipment: Test item/Control Fertiliser: fertiliser spreader for granules (Hege 80 Parzellenstreuer, System Weihenstephan, Model 422437 with a working width of 1.50 m and 10 gape pipes, Hans-Ulrich Hege Saatzuchtmaschinen, Waldenburg, Germany).

Reference item: Movable plot sprayer for field application; type "PSG 4 FE" (Fa. Schachtner Gerätetechnik, 71640 Ludwigsburg, Germany), with an extension tube including 5 spraying nozzles (Lechler IDK 120-04; distance between nozzles: 50 cm) operating with compressed air and a boom width of 250 cm. Distance between the nozzles and soil was approx. 50 cm.

Irrigation: approx. 10 mm irrigation within 3 days after the first application.
Due to a summer characterized by high temperatures and prolonged drought irrigation was performed before the start of the study to prepare suitable sampling conditions (11 mm/m² on August 30, August 31, September 03, September 04 and September 13, 2018 respectively, in total 55 mm/m²).

Collembola Sampling: Soil cores (with subsequent heat extraction) and pitfall traps were used to sample collembola living in the soil and on the soil surface, once before and four times after the application, so far.
>Further samplings are planned for summer and autumn 2019.<

a) Pitfall Traps: Four funnel pitfall traps were installed in the central area of each plot. The traps were opened for 4 days on each sampling occasion and were equipped with a preservative (50 % ethylene glycol, 50 % water) in order to conserve the ground-dwelling arthropods.

b) Heat Extraction of Soil Cores:
Six soil cores per plot were collected with a depth of 5 cm, using a core sampler with 5 cm diameter. The samples were taken from the core area of each plot.
c) Sample Processing: The pitfall trap catches were washed with tap water to remove soil and plant material after sampling; washing water was poured through a sieve (mesh size circa 150 µm). The collected arthropods therefore have a minimum body size of slightly more than 150 µm. Specimens were stored in 70 % ethanol until identification by experts at Mesocosm.
MacFayden heat extraction of soil cores was done by ibacon. Extracted collembola were stored in 70 % ethanol until identification by Mesocosm experts.

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
Endpoints: Abundance and activity of total collembola and abundant collembola species
Statistics: Abundance of collembola was tested for normality (Shapiro Wilks test) and for homogeneity of variance (Levene test) using ToxRat Professional (ToxRat Solutions GmbH, 52477 Alsdorf, Germany, Version 3.3.0, release 20.10.2018).
Depending on the results, Student-t-Test, Welch-t-Test or Mann-Whitney-U-Test was selected to compare control and reference item. These tests were calculated for one-sided smaller (p = 0.05). The difference between means in the control and fertiliser control was calculated the same way but the respective tests were calculated “two-sided “(p=0.05).
To test on significance of the differences between means in controls and treatments for each taxon and sampling date the multiple t-test by Williams (WILLIAMS, 1971, 1972) was used providing the NOEC (No Observed Effect Concentration) on the population level (alpha = 0.05, one-sided). Minimal Detectable Difference (MDD) at the NOEC is also given in accordance with Brock et al. (2015).
For NOEC-calculation abundance data of the taxa were log transformed [y’ = ln (ay+1)] with a = 2/min(x) before the analysis in order to better approximate normality and homoscedasticity (homogeneity of variances) requirements, whereby min(x) was the smallest value of the data set, which was greater than zero.
NOEC-calculations were done with the program Community Analysis (CA) 4.3.14. A previous version of the CA program is described in HOMMEN et al. (1994).


Verification of
Application: Determination of remaining quantity of applicated fertilizer showed that almost 100 % of the test item was applied to each plot.
For determination of variability of distribution five collecting trays (37.5 x 50 cm) were placed on each test item and control fertiliser plot. The trays were passed only once during the application. Weight of the granules in the trays was determined. The applied amount of test item per hectare was calculated on the basis of the surface area of the collection trays and the collected amount of test item.
According to DIN EN 13739-1 (2012) variability of the distribution for three of the four test item plots of each treatment level was “very good” and for one plot “good”.

Nominal and measured concentrations:

1.) 200 kg Perlka® / ha
2.) 400 kg Perlka®/ ha
Perlka® at both rates was applied twice, with an interval of 6 months.

Reference substance (positive control):

yes

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: Abundance

Remarks:

SumCollembola

Remarks on result:

other: pitfall samples compared to untreated control

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

< 200 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: pitfall samples compared to untreated control

Duration:

185 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

Sum/Collembola

Remarks on result:

other: of pitfall samples compared to untreated control

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: pitfall samples compared to fertiliser control

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

200 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: pitfall samples compared to fertiliser control

Duration:

185 d

Dose descriptor:

NOEC

Effect conc.:

200 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: pitfall samples compared to fertiliser control

Duration:

4 d

Dose descriptor:

NOEC

Effect conc.:

< 200 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to untreated control

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to untreated control

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to untreated control

Duration:

4 d

Dose descriptor:

NOEC

Effect conc.:

200 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to fertiliser control (

Duration:

14 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to fertiliser control (

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 400 other: kg Perlka® / ha

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: abundance

Remarks:

SumCollembola

Remarks on result:

other: soil core samples compared to fertiliser control

Details on results:

See "Any other information on results"

Results with reference substance (positive control):

See "Any other information on results"

Reported statistics and error estimates:

See "Any other information on results"

EXPOSURE

For the first application of both test item treatments the exposure could be confirmed by a negligible residue of 0.1 – 0.4 % of the application rate in the fertiliser distributor. Distribution variability was “very good” for three of the four plots and “good” for one plot respectively, according to DIN EN 13739-1 (2012). [DIN EN 13739-1 (2012): Agricultural machinery - Solid fertiliser broadcasters and full width distributors-Environmental protection - Part 1: Requirements]

For the control fertiliser the exposure could also be confirmed by no residual amount after each application in the fertiliser distributor. The lower quality of the distribution variability with CV values between 17.7 % to 22.9 % was probably more a shortcoming of the method then a distribution problem, because of the low amount of fertiliser per square meter, the limited area of the collecting trays and the irregular grain size of the control fertiliser.

The application of the reference item was confirmed by a low deviation between 1.5 to 2.3 % from the target amount.

 

VALIDITY OF THE STUDY

The validity criterion is met for all three post-application sampling dates evaluated for both methods after the first application.

A statistically significant reduction by at least 50 % compared to the untreated control could be observed for six of the eight different species found during the three sampling dates of pitfall traps and soil cores, for which an evaluation was reasonable, thus demonstrating the sensitivity of the test design.

 

EFFECT CLASSIFICATION

Up to date the taxonomic evaluation of pitfall samples of day -3, 7, 14, 28 and 185 and of soil core samples of day -2, 4, 14 and 28 after the first application in autumn 2018 is completed. A second application was done on April 02, 2019 followed by further samplings. Last sampling is planned for autumn 2019. Therefore, the existing results just give hints for the short-term effects of the autumn application in 2018 of Perlka® with the treatment rates of 200 and 400 kg Perlka®/ha for soil cores. In contrast, medium-term effects could already be evaluated for the pitfall traps.

In the pitfall and soil core samples that are taxonomically evaluated up to date 15 different collembola species were determined.

The comparison of control and fertiliser control samples from pitfall traps and soil cores samples revealed several statistically significant differences. Therefore, the fertilizing impact of the control fertiliser seems to have an influence on collembola abundance leading to an independent effect evaluation with respect to untreated control and fertiliser control.

 

PITFALL TRAP SAMPLES

For pitfall trap samples evaluated so far (up to day 185 after the first application) for the comparison to the fertiliser control for both treatment rates only short-term effects could be observed on single sampling dates.

For the comparison to the untreated control up to date also only short-term effects could be observed. Form the data available to date a recovery for Sminthurius aureus at least for the low treatment can be assumed. Due to the sampling early in the year abundance were not yet high enough to show a clear recovery, but values in controls and the low treatment show a potential for recovery.

Thus, in conclusion results available up to date show that for both sampling techniques compared to the fertiliser control only short-term effects followed by recovery could be observed. For comparison to the untreated control effects were also only short-term which has to be proven for S. aureus and Isotoma viridis by further data of the samplings in spring and autumn 2019.

 

Table 1 and 2 show results of the NOEC calculation. +/ - indicates a significantly higher or lower mean abundance compared to control, e.g. NOEC “< 200-“ means abundances in 400 and 200 kg/ha treatment level are significantly lower compared to control.

Tab.1: NOEC and MDD-values (in brackets) for statistical evaluation of pitfall samples compared to untreated control (Williams-test, one-sided, α = 0.05, significant effects bold).

day	-3	7	14	28	185
SumCollembola	≥ 400 (45)	≥ 400 (35)	≥ 400 (40)	< 200- (31)	≥ 400 (44)
Isotoma viridis	≥ 400 (114)	≥ 400 (48)	≥ 400 (93)	≥ 400 (91)	200- (70)
Lepidocyrtus violaceus	≥ 400 (45)	≥ 400 (36)	≥ 400 (43)	200- (35)	≥ 400 (47)
Lepidocyrtus lignorum	≥ 400 (46)	≥ 400 (43)	≥ 400 (48)	≥ 400 (31)	≥ 400 (64)
Heteromurus nitidus	≥ 400 (141)	≥ 400 (87)	≥ 400 (102)	≥ 400 (93)	≥ 400 (121)
Sminthurinus aureus	≥ 400 (96)	≥ 400 (64)	200- (88)	< 200- (77)	≥ 400 (110)
Sminthurus viridis	≥ 400 (n.c.)	≥ 400 (138)	≥ 400 (107)	≥ 400 (261)	≥ 400 (134)
Folsomia manolachi	≥ 400 (n.c.)	≥ 400 (n.c.)	≥ 400 (139)	200+ (145)*
Orchesella flavescens	≥ 400 (n.c.)	≥ 400 (121)	≥ 400 (152)	≥ 400 (103)	≥ 400 (109)
Pseudosinella alba		≥ 400 (n.c.)
Neanura muscorum		≥ 400 (n.c.)		≥ 400 (n.c.)
Ceratophysella sp.
Parisotoma notabilis				≥ 400 (n.c.)
Lepidocyrtus juvenil

n.c. = not calculated due to 0 abundance in control, * not relevant due to low abundance values.

 

Tab.2: NOEC and MDD-values (in brackets) for statistical evaluation of pitfall samples compared to fertiliser control (Williams-test, one-sided, α = 0.05, significant effects bold).

day	-3	7	14	28	185
SumCollembola	≥ 400 (45)	≥ 400 (30)	≥ 400 (45)	200- (31)	200+ (45)
Isotoma viridis	≥ 400 (126)	≥ 400 (65)	≥ 400 (98)	≥ 400 (98)	≥ 400 (75)
Lepidocyrtus violaceus	≥ 400 (46)	≥ 400 (31)	≥ 400 (47)	200- (37)	200+ (49)
Lepidocyrtus lignorum	≥ 400 (39)	≥ 400 (42)	≥ 400 (53)	< 200- (28)	≥ 400 (66)
Heteromurus nitidus	≥ 400 (164)	≥ 400 (72)	≥ 400 (100)	≥ 400 (107)	≥ 400 (n.c.)
Sminthurinus aureus	≥ 400 (89)	≥ 400 (79)	≥ 400 (92)	≥ 400 (81)	≥ 400 (150)
Sminthurus viridis	≥ 400 (198)	≥ 400 (141)	≥ 400 (186)	≥ 400 (111)
Folsomia manolachi	≥ 400 (n.c.)	≥ 400 (n.c.)	≥ 400 (n.c.)	200+ (117)*	≥ 400 (130)
Orchesella flavescens	≥ 400 (n.c.)	< 200- (63)*	≥ 400 (191)	≥ 400 (n.c.)	≥ 400 (120)
Pseudosinella alba		≥ 400 (n.c.)
Neanura muscorum		≥ 400 (n.c.)		≥ 400 (n.c.)
Ceratophysella sp.	≥ 400 (146)	≥ 400 (146)
Parisotoma notabilis				≥ 400 (n.c.)
Lepidocyrtus juvenil	≥ 400 (139)

 

n.c. = not calculated due to 0 abundance in control, * not relevant due to mean abundance of < 1 in fertiliser control,

 

b) Heat Extraction of Soil Cores:

For soil core samples evaluated up to date (day 28 after the first application) only short-term effects (total abundance and Lepidocyrtus violaceus) on single sampling days down to the low concentration could be observed. It has to be considered that both effects show no clear dose-response relationship and are followed by clear recovery.

 

Tab.3: NOEC and MDD-values (in brackets) of soil core samples compared to untreated control (Williams-test, one-sided, α = 0.05, significant effects bold).

day	-2		4	14	28
Sum collembola	≥ 400 (23)		< 200- (25)	≥ 400 (26)	≥ 400 (20)
Lepidocyrtus violaceus	≥ 400 (27)		< 200- (27)	≥ 400 (29)	≥ 400 (29)
Lepidocyrtus lignorum	≥ 400 (90)		≥ 400 (96)	≥ 400 (95)	≥ 400 (97)
Parisotoma notabilis	≥ 400 (100)		≥ 400 (83)	≥ 400 (97)	≥ 400 (64)
Folsomia manolachi	≥ 400 (90)		≥ 400 (90)	≥ 400 (92)	≥ 400 (57)
Isotoma viridis	≥ 400 (93)		≥ 400 (73)	≥ 400 (63)	≥ 400 (88)
Protaphorura armata	≥ 400 (159)		≥ 400 (96)	≥ 400 (134)	≥ 400 (96)
Frisea mirabilis				≥ 400 (156)	≥ 400 (n.c.)
Sminthurinus aureus	≥ 400 (84)		200- (82)*	≥ 400 (114)	200+ (246)*
Lepidocyrtus juvenil			≥ 400 (n.c.)
Tullbergia simplex	≥ 400 (141)		≥ 400 (92)	≥ 400 (95)	≥ 400 (114)
Heteromurus nitidus			≥ 400 (221)	≥ 400 (113)	≥ 400 (176)

 

n.c.= not calculated due to no abundances in control, * not relevant due to low abundance,

 

Tab.4: NOEC and MDD-values (in brackets) of soil core samples compared to fertiliser control (Williams-test, one-sided, α = 0.05, significant effects bold).

day	-2	4	14	28
Sum collembola	≥ 400 (23)	200+ (30)	≥ 400 (26)	≥ 400 (24)
Lepidocyrtus violaceus	≥ 400 (27)	≥ 400 (39)	≥ 400 (27)	≥ 400 (35)
Lepidocyrtus lignorum	≥ 400 (98)	≥ 400 (101)	≥ 400 (100)	≥ 400 (105)
Parisotoma notabilis	≥ 400 (105)	≥ 400 (97)	≥ 400 (98)	≥ 400 (60)
Folsomia manolachi	≥ 400 (98)	200+ (97)	≥ 400 (70)	≥ 400 (62)
Isotoma viridis	≥ 400 (90)	≥ 400 (73)	≥ 400 (64)	≥ 400 (88)
Protaphorura armata	≥ 400 (159)	≥ 400 (109)	≥ 400 (117)	≥ 400 (90)
Frisea mirabilis	≥ 400 (132)	≥ 400 (126)	≥ 400 (136)	≥ 400 (n.c.)
Sminthurinus aureus	≥ 400 (98)	≥ 400 (144)	≥ 400 (165)	≥ 400 (102)
Lepidocyrtus juvenil		≥ 400 (167)
Tullbergia simplex	≥ 400 (115)	≥ 400 (116)	≥ 400 (130)	≥ 400 (n.c.)
Heteromurus nitidus		≥ 400 (n.c.)	≥ 400 (115)	≥ 400 (97)

 

n.c.= not calculated due to no abundances in fertiliser control,

 

DISCUSSION AND CONCLUISON:

Results available up to date show that compared to the fertiliser control only short-term effects followed by recovery could be observed for both sampling techniques.

For comparison to the untreated control effects were also only short-term. Recovery has to be demonstrated for Sminthurinus aureus (effects on day 14 and 28 in pitfall samples) and Isotoma viridis (effect on day 185 in pitfall samples) by further data of the samplings in spring and autumn 2019.

 

Validity criteria fulfilled:

yes

Conclusions:

For both sampling techniques application of 200 and 400 kg PERLKA/ha compared to the fertiliser control showed single short-term effects followed by recovery.
When comparing test item findings to the untreated control statistically significant differences were observed only for single species. For an overall evaluation of these differences further data of the samplings in spring and autumn 2019 will complete the picture.
For the assessment of the observed differences to the two different control treatments it has to be taken into account, that fertilising itself has an influence on soil parameters and habitat structure.
Based on the data available up to now there is no evidence that collembola are adversely affected in the longer term by the test item PERLKA at application rates of 200 and 400 kg/ha.

Executive summary:

The aim of this study was to investigate possible side-effects of the mineral fertiliser Perlka® containing 45 % of calcium cyanamide on populations of collembola in the field by performing a field experiment lasting approximately one year with five treatment groups:

·      The first group of plots was treated with 200 kg Perlka®/ha on September 28, 2018 and April 2, 2019.

·      The second group was treated with 400 kg Perlka®/ha on the same days

·      The third group serving as fertilizer control, was treated with the standard nitrogen fertiliser urea Piagran®46 (SKW Piesteritz, total nitrogen content approx. 46.5 % (nominal)) at the same total nitrogen rate as provided by the 400 kg/ha Perlka group, corresponding to 172.9 kg Piagran®46/ha. Applications were done on September 28, 2018 and April 2, 2019.

·      The fourth group was conducted as a negative control without any nitrogen fertiliser treatment.

·      The fifth group was treated with the reference item Agriclor (480 g/L chlorpyrifos) with 0.72 kg chlorpyrifos/ha in 400 L tap water/ha once on September 28, 2018. Chlorpyrifos is known as a substance harmful to collembola (reference).

Possible effects of the test item on the abundances and activity densities of natural collembola populations living in the investigated area were determined and compared between the five different treatment groups.

A hay meadow was selected as a model system due to its diverse and abundant soil fauna community resulting from a lack of soil disturbance, thus enabling the study to cover a wide range of taxa, with sufficient numbers of individuals to allow meaningful statistical analysis. Hence, results can be extrapolated to other cropping systems.

Application of the test item was verified plot by plot by weighing the distribution density of the granules.

 

Up to date the taxonomic evaluation of pitfall samples of day -3, 7, 14, 28 and 185 and of soil core samples of day -2, 4, 14 and 28 after the first application in autumn 2018 is completed. A second application was done on April 02, 2019 followed by further samplings. Last sampling is planned for autumn 2019. Therefore, the existing results just give hints for the short-term effects of the autumn application in 2018 of Perlka® with the treatment rates of 200 and 400 kg Perlka®/ha for soil cores. In contrast, medium-term effects could already be evaluated for the pitfall traps.

In the pitfall and soil core samples that are taxonomically evaluated up to date 15 different collembola species were determined.

Results available up to date show that compared to the fertiliser control only short-term effects followed by recovery could be observed for both sampling techniques.

For comparison to the untreated control effects were also only short-term. Recovery must be demonstrated for Sminthurinus aureus (effects on day 14 and 28 in pitfall samples) and Isotoma viridis (effect on day 185 in pitfall samples) by further data of the samplings in spring and autumn 2019.

Description of key information

The toxicity of calcium cyanamide as well as its degradation product cyanamide to terrestrial arthropods was investigated in different studies on a variety of terrestrial arthropod species. 

There was one study with calcium cyanamide with the carabid beetle, Bemdidion lampros. In addition, there were 14 studies with the degradation product, cyanamide, conducted with nine different species of terrestrial arthropod.  

In the test with PERLKA (a formulation of calcium cyanamide), the test substance was not found to be toxic to the carabid beetle Bembidion lampros up to a soil surface concentration of 450 kg/ha (NOEC) (Mead-Briggs, 1990). Under the assumption of a maximum soil depth of 5 cm (-> soil surface application) and the density of 1500 kg/m³ dry soil, the application rate of 450 kg PERLKA/ha corresponds to a soil concentration of 600 mg PERLKA/kg soil dw, or 264 mg calcium cyanamide/kg soil dw.

In tests with the transformation product cyanamide, the lowest LR50 values were found from the dose-response tests with Typhlodromus pyri (Goßmann, 2000) and Aphidius rhoplalosiphi Moll, 2001). The LR50 values determined were 446 g pure cyanamide/ha (equivalent to 872 g test substance/ha) and 432 g pure cyanamide/ha (equivalent to 856 g test substance/ha), respectively.

In a dose-response test with cyanamide investigating the effects on the springtail Folsomia candida, the EC10 for reproduction was determined to be 1.515 mg pure cyanamide/ka soil dw (Moser, 2009a). This value represents the chronic worst-case value for terrestrial arthropods and is carried forward in the chemical safety assessment.

(A second non-GLP study evaluating the toxicity of cyanamide on the reproduction of Folsomia candida is available (Moser, 2009b). However, the test was conducted with three concentrations only and is thus considered as supporting information.)

For detailled description where read across is used/recommended and where it is preferrable to refain from read across, please see section 13.2 "read across justification for environmental endpoints" and "Scientific rationale for not using cyanamide as read-across substance for calcium cyanamide on toxicological endpoints"

Key value for chemical safety assessment

Long-term EC10, LC10 or NOEC for soil dwelling arthropods:

1.515 mg/kg soil dw

Additional information

Calcium cyanamide:

PERLKA (a formulation of calcium cyanamide) was not found to be toxic to Bembidion lampros up to a soil surface concentration of 450 kg/ha (NOEC) (Mead-Briggs, 1990). Under the assumption of a maximum soil depth of 5 cm (-> soil surface application) and the density of 1500 kg/m³ dry soil, the application rate of 450 kg PERLKA/ha corresponds to a soil concentration of 600 mg PERLKA/kg soil dw, or 264 mg calcium cyanamide/kg soil dw.

The toxicity endpoint value is summarised in the following table:

Species	Test	Measurement endpoint	Endpoint value
Bembidion lampros	acute laboratory test	NOEC mortality	450 kg/ha test substance

 

Possible side-effects of the mineral fertiliser Perlka® containing 45 % of calcium cyanamide on populations of collembola in the field were assessed by Stegger (2019) by performing a field experiment lasting approximately one year with five treatment groups:

·      The first group of plots was treated with 200 kg Perlka®/ha on September 28, 2018 and April 2, 2019.

·      The second group was treated with 400 kg Perlka®/ha on the same days

·      The third group serving as fertilizer control, was treated with the standard nitrogen fertiliser urea Piagran®46 (SKW Piesteritz, total nitrogen content approx. 46.5 % (nominal)) at the same total nitrogen rate as provided by the 400 kg/ha Perlka group, corresponding to 172.9 kg Piagran®46/ha. Applications were done on September 28, 2018 and April 2, 2019.

·      The fourth group was conducted as a negative control without any nitrogen fertiliser treatment.

·      The fifth group was treated with the reference item Agriclor (480 g/L chlorpyrifos) with 0.72 kg chlorpyrifos/ha in 400 L tap water/ha once on September 28, 2018. Chlorpyrifos is known as a substance harmful to collembola (reference).

Possible effects of the test item on the abundances and activity densities of natural collembola populations living in the investigated area were determined and compared between the five different treatment groups.

A hay meadow was selected as a model system due to its diverse and abundant soil fauna community resulting from a lack of soil disturbance, thus enabling the study to cover a wide range of taxa, with sufficient numbers of individuals to allow meaningful statistical analysis. Hence, results can be extrapolated to other cropping systems.

Application of the test item was verified plot by plot by weighing the distribution density of the granules.

Up to date the taxonomic evaluation of pitfall samples of day -3, 7, 14, 28 and 185 and of soil core samples of day -2, 4, 14 and 28 after the first application in autumn 2018 is completed. A second application was done on April 02, 2019 followed by further samplings. Last sampling is planned for autumn 2019. Therefore, the existing results just give hints for the short-term effects of the autumn application in 2018 of Perlka® with the treatment rates of 200 and 400 kg Perlka®/ha for soil cores. In contrast, medium-term effects could already be evaluated for the pitfall traps.

In the pitfall and soil core samples that are taxonomically evaluated up to date 15 different collembola species were determined.

Results available up to date show that compared to the fertiliser control only short-term effects followed by recovery could be observed for both sampling techniques.

For comparison to the untreated control effects were also only short-term. Recovery must be demonstrated for Sminthurinus aureus (effects on day 14 and 28 in pitfall samples) and Isotoma viridis (effect on day 185 in pitfall samples) by further data of the samplings in spring and autumn 2019.

 

Cyanamide:

In addition to the above study, there were 14 available short and long-term studies with the transformation product cyanamide. The corresponding toxicity endpoint values are summarised in the following table:

 

Species	Test	Measurement endpoint	Endpoint value
			Test substance	Active ingredient
Apis mellifera	Oral contact (overspray with 0.2 L product/L)	LD50	<100 µg test substance/bee	<51.6 µg pure cyanamide/bee Abnormal behaviour and mortality
Aphidius rhopalosiphi	acute laboratory test	mortality	100% at 39.14 kg test substance/ha	100% at 20 kg pure cyanamide/ha
Aphidius rhopalosiphi	dose-response test	mortality	LR50 = 0.856 kg test substance/ha	LR50 = 0.432 kg pure cyanamide/ha
Aphidius rhopalosiphi	dose-response test	mortality	LR50 = 1.21 kg test substance/ha	LR50= 0.620 kg pure cyanamide/ha
Chrysoperla carnea	acute laboratory test	mortality	92%	92%
Chrysoperla carnea	dose-response test	mortality	LR50 = 6.65 kg test substance/ha	LR50= 3.4 kg pure cyanamide/ha
Aleochara bilineata	dose-response test	reproduction	ER50 1.82 kg test substance/ha	ER50= 0.93 kg pure cyanamide/ha
Aleochara bilineata	acute laboratory test	reproduction	99.7% at 39.14 kg test substance/ha	99.7% at 20 kg pure cyanamide/ha
Aleochara bilineata	extended laboratory test	reduction of reproduction	15.6%	15.6 %
Poecilus cupreus	acute laboratory test	mortality/reduction of feeding rate	1.4 %	1.4 %
Pardosa spec.	acute laboratory test	mortality/reduction of feeding rate	100% at 39.14 kg test substance/ha	100% at 20 kg pure cyanamide/ha
Pardosa spec.	dose-response test	mortality	LR50 = 4.1 kg test substance/ha	LR50 = 2.1 kg pure cyanamide/ha
Typholodromus pyri	dose-response test	mortality	LR50 = 0.872 kg test substance/ha	LR50 = 0.446 kg pure cyanamide/ha
Typholodromus pyri	dose-response test	mortality	LR50 = 4.04 kg test substance/ha	LR50 = 2.07 kg pure cyanamide/ha
Folsomia candida	dose-response test	reproduction		EC10 = 1.515 mg pure cyanamide/kg soil dw (Moser, 2009a) EC50 = 61.7 mg pure cyanamide/kg soil dw (Moser, 2009b)

 

There are two 28-day long-term toxicity studies with the soil-dwelling rove beetle, Aleochara bilineata. Drexler (2000) exposed adult beetles to sand substrate dosed with SKW cyanamide L 500 (51.1% w/w) aqueous solution in a limit test. Reproduction was significantly reduced and the LOEC was determined to be <20 mg ai/kg. In another long-term study (Röhling, 2006) larvae were exposed to a range of spray residue concentrations resulting in a NOEC (reproduction) of 0.59 kg test substance/ha (equivalent to 0.3 kg ai/ha). The ER50, based on mean no. hatched beetles of F1 generation per introduced female, was 1.8 mg test substance/ha (equivalent to 0.93 kg ai/ha).

There are two short-term studies with the green lacewing, Chrysoperla carnea. Röhling (2006) exposed larvae of the green lacewing to spray residues over a period of 18 days. The LR50 (mortality) was determined to be 6.65 kg test substance/ha and the NOEC was 4.89 kg test substance/ha. A second study (Moll, 2000) was a limit test with a single test concentration of 39.14 kg test substance/ha. The LR50 was determined to be <39.14 kg test substance/ha.

There are three tests with the leaf-dwelling parasitic wasp, Aphidius rhopalosiphi. The 14-day long-term test involved exposing the test organism to a range to test concentrations sprayed onto barley plants. The resulting LR50 was determined to be 1.2 mg test substance/ha (equivalent to 0.62 kg ai/ha). The NOEC based on mortality was 0.67 kg test substance/ha (equivalent to 0.34 kg ai/ha and the NOEC based on reproduction was estimated to be ≥ the highest tested concentration of 1.14 kg test substance/ha (equivalent to 0.58 kg ai/ha). 

A 48-hour short-term study (Moll, 2000) was conducted by exposing the test organisms to dried residues of SKW cyanamide L 500 in a limit test. There was 100% mortality observed at the single test concentration, therefore, the LR50 was determined to be <39.14 kg test substance/ha. A further short-term study with the leaf-dwelling parasitic wasp, Aphidius rhopalosiphi (Moll, 2001) was conducted using a range of test concentrations over 48 hours, and the test organisms were exposed to dried spray residue. The LR50 (mortality) was 854 g test substance/ha (equivalent to 432 g ai/ha). 

There are two available studies with the leaf-dwelling predatory mite, Typhlodromus pyri. A 7-day study (Goßmann, 2000) exposed the test organisms to a range of test concentrations and found the LR50 (mortality) was 872 g test substance/ha (equivalent to 446 g ai/ha). Reproduction was found to be significantly reduced at 587 g test substance/ha (300g ai/ha) but not at 1018 g test substance/ha (520 g ai/ha), therefore, the result observed in this test was not dose-related. A second test run over 14-days (Röhling, 2007) exposed the test organisms to spray residue on leaf discs. This study found the LR50 (mortality) was 4.0 kg test substance/ha (equivalent to 2.07 kg ai/ha). Effects on reproduction were <50% at the highest tested concentration, therefore, the LR50 (reproduction) was considered to be >3.39 kg test substance/ha (>1.73 kg ai/ha).

Five other terrestrial arthropod short-term and long-term studies were available using five different test organisms:

A 28-day long-term study with Folsomia candida (Moser, 2009a) found an EC10 based on reproduction of 1.515 mg pure cyanamide/kg soil dw.

There were two 14-day studies conducted with wolf spiders and a soil-dwelling lycosid spider (Pardosa spp.). The LR50 for wolf spiders was determined to be 2.1 kg ai/ha (Röhling, 2006) and <20 kg ai/ha for the soil-dwelling lycosid spider (Schmitzer, 2000). 

A further 14-day study was conducted with the soil-dwelling beetle, Poecilus cupreus (Schmitzer, 2000) and the test substance was found to have no effect, based on mortality and food consumption, at 20 kg ai/ha. 

A 72-hour feeding study using honey bees (Apis mellifera) determined an oral LD50 of <100 ug test substance/bee and the test item was classified as harmful to bees (Kleiner, 1992).