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Toxicity to soil macroorganisms except arthropods

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toxicity to soil macroorganisms except arthropods: short-term
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Reference Type:
Tri-n-butyl phosphate induced earthworm intestinal damage by influencing nutrient absorption and energy homeostasis of intestinal epithelial cells
Yang Yang, Peng Liu, Mei Lia
Bibliographic source:
Journal of Hazardous Materials 398 (2020) 122850

Materials and methods

Test guideline
equivalent or similar to guideline
OECD Guideline 207 (Earthworm, Acute Toxicity Tests)
Version / remarks:
Version: 1984
GLP compliance:

Test material

Constituent 1
Chemical structure
Reference substance name:
Tributyl phosphate
EC Number:
EC Name:
Tributyl phosphate
Cas Number:
Molecular formula:
tributyl phosphate
Details on test material:
No further information is reported.

Sampling and analysis

Analytical monitoring:

Test substrate

Details on preparation and application of test substrate:
500 mg/L TnBP was ultrasonic dispersed at 50 °C and sequentially dissolved with distilled water, 10 mL different concentrations of 500, 50 and 5 mg/L TnBP were added to 165 mL distilled water and then thoroughly mixed with the artificial soil to obtain desired TnBP concentration (0, 0.1, 1, and 10 mg/kg (dry
soil)). Different concentrations of TnBP (0, 0.1, 1, and 10 mg/kg (dry soil)) were prepared with distilled water and were thoroughly mixed with the artificial soil.

Test organisms

Test organisms (species):
Eisenia fetida
Animal group:
Details on test organisms:
Earthworms cultured outdoor were obtained from a local farming factory (Jurong, China) and acclimatized at 20 ± 1 °C for at least one week prior to avoid interference caused by external environment and help earthworms adapt to the exposed environment. After acclimatization, healthy adult earthworms with clear clitellum (weighing approximately 500−600 mg) were chosen for artificial soil test.

Study design

Study type:
laboratory study
Substrate type:
artificial soil
Limit test:
Total exposure duration:
14 d

Test conditions

Test temperature:
20 ± 1 °C
final soil moisture 35 % (w/w)
Details on test conditions:
Artificial soil tests were conducted in 1 L clear glass bottles with 500 g dry artificial soil (final soil moisture 35 % (w/w)) following the OECD guidelines (OECD, 1984).
After the prepared soil had stabilized at 20 ± 1 °C for one week, 10 earthworms (acclimatized to artificial soil for 24 h) were placed in each container and kept in an incubator at 20 ± 1 °C in the dark for 14 days. Each concentration consisted of six bottles/replicates.
Nominal and measured concentrations:
nominal: 0, 0.1, 1, and 10 mg/kg (dry soil)

Results and discussion

Details on results:
A LC50 value was not reported.

Any other information on results incl. tables

The weight of earthworms increased in all treatments with time and multi way ANOVA identified a significant difference between biomass and time at the 0.05 level; however, no significant dose-related difference were observed between treatments.

Applicant's summary and conclusion

Validity criteria fulfilled:
The mortality in the controls did not exceed 10% at the end of either test.
The artificial soil test showed no significant dose-related difference in mortality (lower than 5%) for the exposure of E. fetida with tributyl phosphate (nominal concentration: 0, 0.1, 1, and 10 mg/kg (dry soil)) over a period of 14 days.
Executive summary:

Artificial soil tests similar to OECD TG 207 suggested that the acute toxicity of tributyl phosphate (TnBP) towards earthworm Eisenia fetida is quite low since no mortality occurred and a steady increase of biomass was observed.

The overall objective of the present study was to investigate the TnBP induced intestinal toxicity mechanism on earthworm Eisenia fetida as well as the potential role of gut bacteria on host’s health. TnBP was found to have high bioconcentrations in earthworm intestinal tract. Digestive tract degradation and digestive enzyme activities disruption associated with nutrients absorption were noticed. Using multi-omics approaches, detailed intestinal toxic mechanism of earthworms under TnBP exposure was provided.
Tight junctions between small intestinal epithelial cells and osmotic equilibrium were destroyed under 10 mg/kg TnBP, leading to nutrient absorption disturbance. To satisfy the excessive energy requirements induced by TnBP, amino acids gluconeogenesis and protein degradation were detected. Moreover, TnBP significantly decreased the diversity of gut microbiota and changed their structure and function involved in hosts’ health and nutrients supply.

The long-term toxicity data was not generated according to the specifications set out in OECD TG222.