PREVALENCE AND GENETIC LOCATION OF NONTRANSFERABLE TRIMETHOPRIM-RESISTANT DIHYDROFOLATE-REDUCTASE GENES IN SOUTH-AFRICAN COMMENSAL FECAL ISOLATES

In a recent survey of trimethoprim resistance, 357 Gram-negative aerobic organisms were isolated from healthy volunteers from rural and urban populations in South Africa. Trimethoprim resistance did not transfer to an Escherichia coli J62-2 recipient strain by conjugation in a liquid mating: in 161 (45.1%) of the isolates. These isolates which did not transfer their resistance were probed with intragenic oligonucleotide probes for the types Ia, Ib, IIIa, V, VI, VII, VIII, IX, X and XII dihydrofolate reductase genes. Contrary to all previous data, the most prevalent dihydrofolate reductase gene in this group of non-transferable isolates which hybridized, was the type VII (38%) followed by the type Ia (25%), Ib (12%), V (1.7%) and VIII (1.2%). None of the strains hybridized to the types IIIa, VI, XI, X and the XII dihydrofolate reductase probes. Southern blots of plasmid and chromosomal DNA from selective isolates revealed that the type VII dihydrofolate reductase genes were located on the chromosome and were associated with the integrase gene of Tn21. However, the type Ib and V dihydrofolate reductase genes were all found on plasmids which could not be mobilized. The type Ia dihydrofolate reductase genes were found on both nontransferable plasmids and on the chromosome. The nature of the genetic structures associated with a dihydrofolate reductase gene strongly affects the means of spread of the gene in a population.