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Applied and Environmental Microbiology, June 1999, p. 2376-2381, Vol. 65, No. 6
Biotechnology Center for Agriculture and the
Environment, Cook College, Rutgers University, New Brunswick, New
Jersey 08901-8520
Received 16 November 1998/Accepted 21 March 1999
Earthworm egg capsules (cocoons) may acquire bacteria from the
environment in which they are produced. We found that Ralstonia eutropha (pJP4) can be recovered from Eisenia 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 of R. 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 containing R. 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 CFU R. eutropha (pJP4)
g
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Earthworm Egg Capsules as Vectors for the
Environmental Introduction of Biodegradative Bacteria
and
1 of soil. The toxicity of 2,4-DCP to developing
earthworms was tested by using cocoons with or without R. eutropha (pJP4). Results showed that cocoons containing R. eutropha (pJP4) were able to tolerate higher levels of 2,4-DCP.
Our results indicate that the biodegradation of 2,4-DCP by R. 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.
*
Corresponding author. Mailing address: Department of
Biochemistry and Microbiology, Cook College, Rutgers University, 76 Lipman Drive, New Brunswick, NJ 08901-8525. Phone: (732) 932-9763. Fax: (732) 932-8965. E-mail: haggblom{at}aesop.rutgers.edu.
Present address: Environmental Molecular Biology, M888, Life
Sciences Division, Los Alamos National Laboratory, Los Alamos, NM 87545.
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