Survival ofluxAB-markedAlcaligenes eutrophus H850 in PCB-contaminated soil and sediment

Abstract

A rifampicin-resistant PCB-degrading Alcaligenes eutrophus H850 strain was marked with luxAB reporter genes and designated H850Lr. This strain was enumerated in soil by viable plating and counting of light-emitting colonies. The marked strain was also inoculated into soil and sediment microcosms contaminated with PCBs and treated with rhamnolipid biosurfactants produced by Pseudomonas aeruginosa UG2Lr or inoculated with the P. aeruginosa UG2Lr strain. A. eutrophus H850Lr exhibited similar survival in sandy loam soil in the absence or presence of PCBs over 56 days. Survival of A. eutrophus H850Lr in PCB-contaminated sediment was less than in sandy soil under the same incubation conditions. Addition of P. aeruginosa UG2 rhamnolipids to soil increased the culturable indigenous heterotrophic population, and numbers of A. eutrophus H850Lr cells. P. aeruginosa UG2Lr cells did not affect survival of A. eutrophus H850, as cell enumerations after 2 months were the same as in microcosms containing only A. eutrophus H850 inoculum. P. aeruginosa UG2Lr survived in soils as demonstrated by the slight decrease in CFU from 1 x 10(8) to 2 x 10(6) CFU cm-3 after 2 months. Direct extraction of DNA from soil and purification for use in PCR amplification using primers specific for the bphC gene detected 8 x 10(2) A. eutrophus H850Lr CFU g-1 soil in PCB-contaminated soils. Colony lifts of bacteria isolated from microcosms containing PCB-contaminated soil did not hybridize with LB400 bphC probe. However, enrichment of PCB-contaminated soil with biphenyl, followed by DNA extraction and probing with bphC gene probe detected indigenous PCB-degrading bacteria containing a similar gene sequence in PCB-contaminated sediment. This study demonstrates the usefulness of using the lux reporter system in monitoring bacterial survival in PCB-contaminated soils and sediments.