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Earthworm egg capsules (cocoons) may acquire bacteria from the environment in which they are produced. We found thatRalstonia eutropha (pJP4) can be recovered fromEisenia fetida cocoons formed in soil inoculated with this bacterium. Plasmid pJP4 contains the genes necessary for 2,4-dichlorophenoxyacetic acid (2,4-D) and 2,4-dichlorophenol (2,4-DCP) degradation. In this study we determined that the presence ofR. eutropha (pJP4) within the developing earthworm cocoon can influence the degradation and toxicity of 2,4-D and 2,4-DCP, respectively. The addition of cocoons containingR. eutropha (pJP4) at either low or high densities (102 or 105 CFU per cocoon, respectively) initiated degradation of 2,4-D in nonsterile soil microcosms. Loss of 2,4-D was observed within the first week of incubation, and respiking the soil with 2,4-D showed depletion within 24 h. Microbial analysis of the soil revealed the presence of approximately 104 CFUR. eutropha (pJP4) g−1 of soil. The toxicity of 2,4-DCP to developing earthworms was tested by using cocoons with or withoutR. eutropha (pJP4). Results showed that cocoons containingR. eutropha (pJP4) were able to tolerate higher levels of 2,4-DCP. Our results indicate that the biodegradation of 2,4-DCP byR. eutropha (pJP4) within the cocoons may be the mechanism contributing to toxicity reduction. These results suggest that the microbiota may influence the survival of developing earthworms exposed to toxic chemicals. In addition, cocoons can be used as inoculants for the introduction into the environment of beneficial bacteria, such as strains with biodegradative capabilities.

Earthworm Egg Capsules as Vectors for the Environmental Introduction of Biodegradative Bacteria