Registration Dossier

Ecotoxicological information

Endpoint summary

Administrative data

Description of key information

Additional information

Toxicity to soil macroorganisms

In a study conducted according to OECD Guideline 222 (Earthworm Reproduction Test (Eisenia fetida/Eisenia andrei)), ISO 11268-2 (Effects of Pollutants on Earthworms. 2. Determination of Effects on Reproduction), and BBA Guideline (Guidelines for the testing of herbicides within registration progress - Effects of herbicides on reproduction and growt of Eisenia fetida/Eisenia andrei; 1994), the influence of Biofert Plusz on mortality, biomass, reproduction, behaviour and pathological symptoms of earthworms was investigated. Forty earthworms (4 replicates: 10 adults each), Eisenia fetida, were exposed to Biofert concentrations ranging from 1.25 to 20 g test material dry matter/kg soil dw for 28 d under static conditions.Thereafter, the adults were removed from the test vessels and mortality and body weight determined. After further 4 weeks the reproduction rate was determined by counting the hatched juvenils. The 28 d NOEC and LOEC based on mortality and body weight were determined to be 10 and 20 g test material dry matter/kg soil dry weight, respectively. The 56 d NOEC and LOEC based on reproduction were determined to be 2.5 and 5 g test material dry matter/kg soil dry weight, respectively. Furthermore, a 56 d EC50 (reproduction)=3.23 g test material dry matter/kg soil dry weight was found. Furthermore, no biological significant symptoms or changes in the behaviour of the earthworms were observed up to a test concentration of 10 g test material dry matter/kg soil dw after 28 d.

Toxicity to terrestrial arthropods

In Annex X of Regulation (EC) No 1907/2006, it is laid down that long-term toxicity tests with terrestrial arthropods shall be proposed by the registrant if the chemical safety assessment indicates the need to investigate further the effects on terrestrial arthropods. According to Annex I of this regulation, the chemical safety assessment triggers further action when the substance or the preparation meets the criteria for classification as dangerous according to Directive 67/548/EEC or Directive 1999/45/EC or is assessed to be a PBT or vPvB. The hazard assessment of Biofert Plusz reveals neither a need to classify the substance as dangerous to the environment, nor is it a PBT or vPvB substance, nor are there any further indications that the substance may be hazardous to the environment. According to reliable study results, Biofert Plusz is considered to be rapidly degraded in the environment and the bioaccumulation potential is regarded to be insignificant. Moreover, the amino acid threonine as the main constituent, is an essential protein constituent in living organisms and so no metabolites are expected to occur that pose a significant risk to living organisms. Therefore, with respect to animal welfare the performance of a long-term toxicity study with terrestrial arthropods is assumed to be not justifiable.

Long-term toxicity to terrestrial plants

The chronic toxicity of the test item Biofert Plusz to two terrestrial plant species was determined over a period of 61 days (oats) and 43 days (turnip) according to DIN ISO 22030:2005. Test item related effects were observed for both plant species at the test concentrations > 8000 mg/kg dw. No emergence was observed for oats at the concentration 20000 mg/kg dw. At the concentration 8000 mg/kg dw the test item caused broad effects on the development of the shoots, resulting in decreased biomass growth and complete inhibition of blossom development. No emergence was observed for turnip at the concentrations 8000 and 20000 mg/kg dw, therefore no evaluation of chronic effects at these concentrations was possible. The concentration of 8000 mg/kg dry weight is much higher than the maximum recommended application rate (1 g liquid product/kg soil corresponding to 0.3 g dry frozen product/kg soil) and acute effects on plants were foreseeable. Although fertilizers have to be applied in high dosages to supply sufficient nutrition for the arable plants, organic and inorganic compounds concentrations of only one or two order of magnitudes higher than the maximum application rate already result in harmful effects on plants and other soil organisms due to osmotic damage. Other guidelines like the acute plant toxicity guideline OECD TG 208 (Seedling emergence and Seedling Growth Test) recommend to test up to a maximum concentration of 1000 mg test substance / kg dry soil for general chemicals and to test crop protection products up to the maximum recommended application rate.

Up to the tested concentration of 3200 mg/kg dw the test item caused no chronic effects on the development and biomass of the husks.

Short-term toxicity to terrestrial plants

Toxicity to terrestrial plants (at pH 4.5)

The phytotoxicity of Biofert Plusz towards terrestrial plants (A. sativa, Glycine max, Sinapis alba) was investigated in a study conducted according to OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test). The seedlings were exposed to 5 test material concentrations (26.9 -2688 mg fresh material/kg soil dw, equivalent to 10 - 1000 mg test material dry matter/kg soil dw) for 21 d at pH 4.5. Shoot height, shoot fresh weight, seedling emergence, and visual phytotoxicity were determined. The 21 d NOEC value for A. sativa in respect to shoot height was determined to be 2688 mg test material/kg soil dw (1000 mg dry matter/kg soil dw) whereas the corresponding value for Sinapis alba was 860 mg test material/kg soil dw (320 mg dry matter/kg soil dw). For Glycine max a 21 d NOEC (shoot height) could not be determined as no concentration effect relationship occurred. The 21 d NOEC values in respect to shoot fresh weight and emerged seedlings were found to be 2688 mg test material/kg soil dw (1000 mg dry matter/kg soil dw) for Avena sativa, Glycine max, and Sinapis alba.Visual phytotoxic effects determined after 7, 14, and 21 d were observed only in a few cases. Lacking of cotyledons (soybean n=1 after 7 d, n=1 after 14 d,and n=1 after 21 d), necrosis (oat n=1 after 14 d; slight effects), dead plants (white mustard n=1 after 14 d, n=1 after 21 d; oat: n=3 after 21 d in two replicates), and stunted growth (white mustard n=1 after 21 d).

Toxicity to terrestrial plants (at pH 4.6)

The phytotoxicity of Biofert Plusz towards terrestrial plants (A. sativa, Glycine max, Sinapis alba) was investigated in a study conducted according to OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test). The seedlings were exposed to 5 test material concentrations (253 -2532 mg fresh material/kg soil, eqiovalent to 10 -1000 mg dry matter/kg soil) for or 21 d at pH 4.6.

Shoot height, shoot fresh weight, seedling emergence, and visual phytotoxicity were determined. The 21 d NOEC value for A. sativa in respect to shoot height was determined to be 2532 mg test material/kg soil dw (1000 mg dry matter/kg soil dw) whereas the corresponding value for Sinapis alba was 810 mg test material/kg soil dw (320 mg dry matter/kg soil dw). For Glycine max a 21 d NOEC (shoot height) could not be determined as no concentration effect relationship occurred. The 21 d NOEC values in respect to shoot fresh weight and emerged seedlings were found to be 2532 mg test material/kg soil dw (1000 mg dry matter/kg soil dw) for Avena sativa and Glycine max; for Sinapis alba the 21 d NOEC (shoot fresh weight) was found to be 1468 mg test material/kg soil dw (580 mg dry matter/kg soil dw) and the 21 NOEC in respect to emerged seedlings 2532 mg test material/kg soil dw (1000 mg dry matter/kg soil dw).Visual phytotoxic effects were determined after 7, 14, and 21 d.Only in a few cases phytotoxic effects (stunted growth and dead plants) were observed using oat as test material. Phytotoxic effects like lacking of cotyledons and dead plants were determined during the testing of soybean. Based on the visual observations, white mustard exposed to Biofert Plusz (pH 4.6) showed the highest sensitivity for phytotoxic effects (stunted growth, dead plants).

Toxicity to terrestrial plants (at pH 5.4)

The phytotoxicity of Biofert Plusz towards terrestrial plants (A. sativa, Glycine max, Sinapis alba) was investigated in a study conducted according to OECD Guideline 208 (Terrestrial Plants Test: Seedling Emergence and Seedling Growth Test). The seedlings were exposed to 5 test material concentrations (27.2 -2725 mg/kg soil dw, equivalent to 10 -1000 mg dry matter/kg soil dw).) for 21 d at pH 5.4. Shoot height, shoot fresh weight, seedling emergence, and visual phytotoxicity were determined. The 21 d NOEC values (shoot height, shoot fresh weight, and emerged seedlings) for A. sativa, Glycine max, and Sinapis alba were all found to be 2725 mg test material/kg soil dw (1000 mg dry matter/kg soil dw).Visual phytotoxic effects were determined after 7, 14, and 21 d. Phytotoxic effects were observed only once: at a test concentration of 100 mg dry matter/kg soil dw one dead plant (oat) was detected after 21 d.

Toxicity to soil microorganisms

In a study conducted according to OECD Guideline 217 (Soil Microorganisms: Carbon Transformation Test) long-term potential effects of Biofert Plusz on the carbon transformation activity of soil microorganisms was investigated. Biofert Plusz was applied to the soil at test substance concentrations of 19.5, 78.1, 313, 1250, 5000 and 20000 mg/kg soil dw nominal. The treated soils were incubated for 28 d at 49.9 -53.8% of the maximum water holding capacity, a pH of 6.34 -6.93 and a temperature of 19 -21°C. The glucose induced respiration was measured on day 0 and after 7, 14, and 28 d. On each occassion, aliquots of the soil were amended with glucose (4000 mg/kg soil). Oxygen consumption was measured over the subsequent 24 h period and compared to that of the untreated control.

Biofert Plusz induced a strong increase of the soil respiration rate throughout the first seven days of exposure at 1250, 5000 and 20000 mg/kg soil dw, presumably due to serving as microbial N-source. After further seven days this induction of microbial respiration was down-regulated, resulting in no statistically significant differences compared to the control at all test substance concentrations. After 14 and 28 days of exposure, substrate induced microbial respiration rate at substance concentrations of 313, 1250, 5000 and 20000 mg/kg soil dw leveled down to the value range measured at day 0. Even the highest applied test substance concentration of 20000 mg/kg dw caused no decrease, but instead a slight increase of the glucose induced respiration rate compared to control at day 28. The decrease seen at TS concentrations of 1250 and 5000 mg/kg dw in the course of exposure may be attributable to prolonged adaptation of exposed microbial communities to differing feed availability, resp. changing C/N ratio. Overall results support the conclusion that any long-term effects of the test substance on exposed microbial soil organisms are not to be expected (NOEC > 20000 mg/kg dw).

Toxicity to birds

In Annex X of Regulation (EC) No 1907/2006, it is laid down that chronic toxicity tests in birds shall be proposed by the registrant if the chemical safety assessment indicates the need to investigate further the effects on birds. According to Annex I of this regulation, the chemical safety assessment triggers further action when the substance or the preparation meets the criteria for classification as dangerous according to Directive 67/548/EEC or Directive 1999/45/EC or is assessed to be a PBT or vPvB. The hazard assessment of Biofert Plusz reveals neither a need to classify the substance as dangerous to the environment, nor is it a PBT or vPvB substance, nor are there any further indications that the substance may be hazardous to the environment. According to reliable study results, Biofert Plusz is considered to be rapidly degraded in the environment and the bioaccumulation potential is regarded to be insignificant and therefore secondary poisoning is unlikely to be a relevant exposure route. Moreover, the amino acid threonine as the main constituent, is an essential protein constituent in living organisms and so no metabolites are expected to occur that pose a significant risk to living organisms. Therefore, with respect to animal welfare the performance of a long-term toxicity study with birds is assumed to be not justifiable.