Gene transfer and plasmid instability within pilot-scale sewage filter beds and the invertebrates that live in them

The environmental plasmid pQKH6 was transferred conjugatively between strains of Pseudomonas putida at mean frequencies of up to 8.4x10(-4) within pilot-scale sewage filter beds. This frequency was 10-fold higher than that reported previously for this environment and was probably due to seasonal temperature changes. Many (45%) of the plasmids isolated subsequently from the filter beds had restriction fragment length polymorphism (RFLP) profiles that differed from that expected for pQKH6. RFLP analysis revealed structural rearrangements occurring within a particularly restriction-site-rich region of the plasmid. Although no evidence was obtained showing the indigenous invertebrate populations within the filter beds to influence the rate of gene transfer, pQKH6 was transferred with frequencies of up to 1.6x10(-2) within the guts of the filter-bed-dwelling Sylvicola fenestralis larvae during laboratory experiments. This transfer was strongly influenced by donor to recipient ratios. Laboratory experiments also showed that Serratia fonticola survived better within invertebrate guts than P. putida. This evidence, along with experiments showing that S. fonticola could participate in pQKH6 transfer within filter-bed biofilm, identify this bacterium as a better model than P. putida for examining the effect of invertebrates on gene transfer.