Evaluation of the effect of transgenic Bt cotton on snails Bradybaena (Acusta) ravida and Bradybaena similaris (Ferussac).

The impact of transgenic crops on non-target organisms is a key aspect of environmental safety assessment to transgenic crops. In the present study, we fed two snail species, Bradybaena (Acusta) ravida (B. ravida) and Bradybaena similaris (Ferussac)(B. similaris), with the leaves of transgenic Bt cotton Zhong 30 (Z30) and control cotton, its parent line zhong 16 (Z16), to assess the environmental safety of Bt cotton to common non-target organisms in the field. Survival, body weight, shell diameter, helix number, reproduction rate, superoxide dismutase (SOD) activity and Bt protein concentration in snails were monitored in 15 days and 180 days experiments. We also monitored the population dynamics of B. ravida and B. similaris in Z30 and Z16 cotton fields for two successive years. Compared to the snails fed on the control cotton Z16, there was no significant difference in survival, growth, reproduction, and SOD activity on Bt cotton Z30. Bt protein concentrations were significantly between different treatments, and Bt protein residues were only detected in the feces of the Z30 treatment. According to the field data, the number of B. ravida and B. similaris fluctuated considerably across seasons over the entire cotton-growing season; however, there were no significant differences between the Bt and control cotton fields at similar time. As the results showed, in our experiments, Bt cotton Z30 had no adverse effects on the two snail species, both in the laboratory and in the fields.

Application of Box-Behnken experimental design to optimize the extraction of insecticidal Cry1Ac from soil.

A validated method for analyzing Cry proteins is a premise to study the fate and ecological effects of contaminants associated with genetically engineered Bacillus thuringiensis crops. The current study has optimized the extraction method to analyze Cry1Ac protein in soil using a response surface methodology with a three-level-three-factor Box-Behnken experimental design (BBD). The optimum extraction conditions were at 21 °C and 630 rpm for 2 h. Regression analysis showed a good fit of the experimental data to the second-order polynomial model with a coefficient of determination of 0.96. The method was sensitive and precise with a method detection limit of 0.8 ng/g dry weight and relative standard deviations at 7.3%. Finally, the established method was applied for analyzing Cry1Ac protein residues in field-collected soil samples. Trace amounts of Cry1Ac protein were detected in the soils where transgenic crops have been planted for 8 and 12 years.

Dissipation of insecticidal Cry1Ac protein and its toxicity to nontarget aquatic organisms.

The widespread cultivation of Bacillus thuringiensis crops has raised public concerns on their risk to nontarget organisms. Persistence of Cry1Ac protein in soil, sediment and water and its toxicity to nontarget aquatic organisms were determined. The dissipation of Cry1Ac toxin was well described using first order kinetics, with the half-lives (DT50) ranging from 0.8 to 3.2, 2.1 to 7.6 and 11.0 to 15.8 d in soil, sediment and water, respectively. Microbial degradation played a key role in the dissipation of Cry1Ac toxin and high temperature accelerated the processes. Cry1Ac toxin was more toxic to the midge Chironomus dilutus than the amphipod Hyalella azteca, with the median lethal concentration (LC50) of C. dilutus being 155 ng/g dry weight and 201 ng/mL in 10-d sediment and 4-d water bioassays, respectively. While Cry1Ac toxin showed toxicity to the midges, risk of Bt proteins to aquatic nontarget organisms was limited because their environmentally relevant concentrations were much lower than the LC50s.