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RP4 Plasmid Transfer Among Strains of Pseudomonas in a Biofilm Reactor

EPA Grant Number: U914730
Title: RP4 Plasmid Transfer Among Strains of Pseudomonas in a Biofilm Reactor
Investigators: Ehlers, Laura J.
Institution: Johns Hopkins University
EPA Project Officer: Broadway, Virginia
Project Period: January 1, 1995 through January 1, 1996
Project Amount: $102,000
RFA: STAR Graduate Fellowships (1995)
Research Category: Biology/Life Sciences , Academic Fellowships , Fellowship - Microbiology

Description:

Objective:

The objective of this research project is to observe and quantify conjugation between bacteria growing in a biofilm reactor.

Approach:

The reactor was chosen for its experimental flexibility and simple mathematical description. Several environmental parameters were varied to determine their influence on transfer frequencies. Finally, a predictive biofilm model was evaluated for its ability to accurately represent plasmid transfer events.

Expected Results:

Conjugation of the broad host range plasmid RP4 between two strains of Pseudomonas occured in the biofilm reactor at high frequencies. The most important environmental parameter was the shear stress at the biofilm-liquid interface. Conjugation was only observed below a shear of 0.0851 N/m2, corresponding to a laminar flow regime. Increasing temperature from 15°C to 28°C increased conjugation frequencies 10,000-fold. Conjugation frequency was unchanged in experiments conducted with 3.5, 7, and 35 mg/L acetate, although total cell concentration in the biofilm increased as expected. Finally, the addition of a sublethal kanamycin concentration affected both the recipient and transconjugant populations, either by enhancing conjugation or providing a selective advantage for transconjugant growth. These data suggest ways to manipulate environmental parameters to increase or decrease plasmid transfer rates among biofilm bacteria.

Supplemental Keywords:

fellowship, biofilm reactor, environmental parameters, DNA, genetically engineered microorganisms, GEMs, environment, conjugation, bacteria, Pseudomonas. , Ecosystem Protection/Environmental Exposure & Risk, Scientific Discipline, RFA, Biology, Environmental Microbiology, Monitoring/Modeling, Genetics, genetically engineered bacteria, bacteria monitoring, plasmid transfer, biochemistry, model-based analysis, biofilm dynamics

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The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.

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