In situ studies with membrane diffusion chambers of antibiotic resistance transfer in Escherichia coli

Abstract

Coliform bacteria were isolated from raw sewage and sewage effluent-receiving waters and tested for their antibiotic susceptibility patterns and their ability to transfer antibiotic resistance to Escherichia coli K-12 C600. An environmental isolate of E. coli (MA527) capable of transferring antibiotic resistance to C600 was mated, both in vitro and in situ, with an antibiotic-sensitive E. coli environmental isolate (MA728). In situ matings were conducted in modified membrane diffusion chambers, in the degritter tank at the Grant Street (Melbourne, Fla.) sewage treatment facility, and in the sewage effluent-receiving waters in Melbourne, Fla. The transfer frequencies in situ were 3.2 x 10(-5) to 1.0 x 10(-6), compared with 1.6 x 10(-4) to 4.4 x 10(-5) observed in vitro. Transfer was shown to occur in raw sewage but was not detected in the effluent-receiving waters. The presence of a 60-megadalton plasmid species in both donor and transconjugants, but not in the recipients, provided physical evidence for the transfer of antibiotic resistance in situ.