Boron orthophosphate

Toxicity to soil macroorganisms except arthropods

Link to relevant study record(s)

Reference

Endpoint:

toxicity to soil macroorganisms except arthropods: long-term

Type of information:

experimental study

Adequacy of study:

key study

Study period:

14 Apr - 09 Jun 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei))

GLP compliance:

yes (incl. QA statement)

Remarks:

Ministerium für Arbeit, Integration und Soziales des Landes Nordrhein-Westfalen

Analytical monitoring:

no

Vehicle:

no

Details on preparation and application of test substrate:

- Method of mixing into soil (if used): As the test item is insoluble in water and easily evaporating organic solvents, the quantity of test item required to obtain the desired concentrations was applied to quartz sand by grinding. The spiked quartz sand was distributed on the soil and mixed thoroughly. The test substrate was prorated to each test container.
- Controls: yes

Test organisms (species):

Eisenia andrei

Animal group:

annelids

Details on test organisms:

TEST ORGANISM
- Source: Regenwurmfarm Tacke, Borken, Germany
- Age at test initiation (mean and range, SD): 2 - 12 months
- Weight at test initiation (mean and range, SD): 250 - 600 mg

ACCLIMATION
- Acclimation period:6 days
- Acclimation conditions (same as test or not): The worms were conditioned in the artificial soil before use. The same food which was also used in the test was given in a sufficient amount.

Study type:

laboratory study

Substrate type:

artificial soil

Limit test:

no

Total exposure duration:

56 d

Test temperature:

18 - 22°C

pH:

test start: 6.2 - 6.5
test end: 5.9 - 6.2

Details on test conditions:

TEST SYSTEM
- Test container (material, size): plastic containers (area of 17x12 cm, height of 6 cm); containers were covered with perforated transparent plastic caps
- Amount of soil or substrate: 500 - 650 g dry mass of artificial soil
- No. of organisms per container (treatment): 10
- No. of replicates per treatment group: 4
- No. of replicates per control: 8
- other: One day after addition of the worms, an amount of 5 g air dried finely ground cow manure per test container was spread on the soil surface and moisten with water. Once a week, during the first four weeks, the adult earthworms were fed according to their food consumption.

SOURCE AND PROPERTIES OF SUBSTRATE (if soil)
- Composition (if artificial substrate): Sphagnum peat, air-dried, finely ground 10 %, Kaolinite, air-dried 20 %, Industrial quartz sand, air-dried 70 %
- Pretreatment of soil: The test substrate was wetted with deionised water to a water content of between 40 % and 60 % by mass of the total water-holding capacity. The pH of the substrate was adjusted to 6.0 ± 0.5 (1 mol/L KCl) by adding pulverized calcium carbonate (CaCO3).

OTHER TEST CONDITIONS
- Photoperiod: light/dark cycle 16 h : 8 h
- Light intensity: 732 - 764 lx

EFFECT PARAMETERS MEASURED (with observation intervals if applicable): mortality and weight change after 28 days, offspring number after 56 days

Nominal and measured concentrations:

Nominal: control, 62.5, 125, 250, 500 and 1000 mg TI/kg dry mass soil

Reference substance (positive control):

yes

Remarks:

carbendazim

Duration:

56 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

>= 1 000 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

28 d

Dose descriptor:

NOEC

Effect conc.:

500 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: weight change

Duration:

28 d

Dose descriptor:

EC0

Effect conc.:

750 mg/kg soil dw

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

other: weight change

Details on results:

- Mortality at end of exposure period: ≤ 5%
- Total mass of adults at beginning of test: see "Any other information on results"
- No. of offspring produced: see "Any other information on results"

Results with reference substance (positive control):

- Results with reference substance valid?
- Relevant effect levels: EC10 = 0.9 mg/kg for weight change; EC10 = 2.0 mg/kg for reproduction (results for reproduction are in agreement with the range of 1 - 5 mg carbendazim per kg (OECD 222); weight change and mortality are in agreement with experience in test facikity)
- Other:

Living earthworms after 28 days. Single values of the parallel test pots [Individuals].

TI = Test item; DM = Dry mass; Concentrations given as nominal values.

	replicate	Control	62.5 mg TI per kg DM	125 mg TI per kg DM	250 mg TI per kg DM	500 mg TI per kg DM	1000 mg TI per kg DM
Living earthworms [Ind.]	1	9	10	9	10	10	9
	2	10	9	10	10	9	10
	3	10	10	10	9	10	10
	4	9	10	9	10	10	9
	5	10
	6	10
	7	10
	8	8

 

Biomass of the earthworm batches at test start and after 28 days.Single values of the parallel test pots [g FM].

         TI = Test item; FM = Fresh mass; DM = Dry mass; Concentrations given as nominal values.

	replicate	Control	62.5 mg TI per kg DM	125 mg TI per kg DM	250 mg TI per kg DM	500 mg TI per kg DM	1000 mg TI per kg DM
Test start	1	3.588	3.062	3.131	3.187	3.408	3.752
	2	3.302	2.977	3.554	3.295	2.962	3.313
	3	3.064	3.257	3.378	3.069	3.002	3.327
	4	2.965	3.408	3.272	3.080	3.076	3.426
	5	3.441
	6	3.364
	7	3.409
	8	3.524
28 d	1	4.885	4.917	4.696	4.911	5.098	3.898
	2	4.995	4.063	5.237	5.247	4.227	4.364
	3	4.664	4.749	5.251	4.325	4.839	4.684
	4	4.211	4.966	4.652	4.801	4.692	4.152
	5	5.223
	6	5.061
	7	5.267
	8	4.175

 

Weight change at test end. Weight change compared to the start value [%] and compared to the control [%].

TI = Test item; DM = Dry mass; *: significant when compared with control (Williams test, one-sided); Concentrations given as nominal values.

Test species		Control		62.5 mg TI per kg DM		125 mg TI per kg DM		250 mg TI per kg DM		500 mg TI per kg DM		1000 mg TI per kg DM
compared to start value [%]		45		47		49		53		52		24
compared to control [%]				6		9		18		15		-45 *

 

Reproduction. Single values of the parallel test pots [Individuals]and mean values at the end of the study (56 d) [Individuals].

TI = Test item; DM = Dry mass; CV = Coefficient of variation; Concentrations given as nominal values.

	Replicate	Control	62.5 mg TI per kg DM	125 mg TI per kg DM	250 mg TI per kg DM	500 mg TI per kg DM	1000 mg TI per kg DM
Juveniles [Ind.]	1	120	142	79	120	112	94
	2	116	140	87	112	89	57
	3	127	143	143	48	134	126
	4	48	129	82	83	114	111
	5	97
	6	69
	7	118
	8	129
Mean Juveniles [Ind] (CV)		103 (28.8)	139 (4.7)	98 (31.0)	91 (36.0)	112 (16.4)	97 (30.6)

 

 

Validity criteria fulfilled:

yes

Additional information

One experimental study was conducted investigating the effects of Boron orthophosphate (CAS 13308-51-5) on mortality, body weight and reproduction of Eisenia andrei. Until now preliminary results are available.

The test was performed according to OECD 222 with nominal concentrations of 62.5, 125, 250, 500 and 1000 mg/kg. After 4 weeks mortality of adults was ≤ 5%, resulting in a NOEC ≥ 1000 mg/kg soil dw. For weight change a NOEC (4 wks) of 500 mg/kg soil dw was calculated. After an additional 4 weeks the influence of the test substance on reproductive output of parent animals was recorded. No effects on the number of juveniles were found resulting in a NOEC (8 wks) ≥ 1000 mg/kg soil dw. Thus, the relevant endpoints mortality and reproduction did not result in any effects up to a nominal concentration of 1000 mg/kg soil dw.