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M.L. Krumme

Publications and source records attributed to M.L. Krumme.

3 recordsLinked to original sources

Behavior of pollutant-degrading microorganisms in aquifers: Predictions for genetically engineered organisms

Bioremediation via environmental introductions of degradative microorganisms requires that the microbes survive in substantial numbers and effect an increase in the rate and extent of pollutant removal. Combined field and microcosm studies were used to assess these abilities for laboratory-grown bacteria. Following introduction into a contaminated aquifer, viable cells of Pseudomonas sp. B13 were present in the contaminant plume for 447 days; die-off was rapid in pristine areas. In aquifer microcosms, survival of B13 and FR120, a genetically engineered derivative of B13 having enhanced catabolic capabilities for substituted aromatics, was comparable to B13 field results; both bacteria degraded target pollutants in microcosms made with aquifer samples from the aerobic zone of the pollutant plume. Results suggest that field studies with nonrecombinant microorganisms may be coupled to laboratory studies with derivative strains to estimate their bioremediative efficacy. Furthermore, laboratory strains of bacteria can survive for extended periods of time in nature and thus may have important bioremediative applications.

Environmental Science & Technology↗

Use of molecular techniques to evaluate the survival of a microorganism injected into an aquifer

A PCR primer set and an internal probe that are specific for Pseudomonas sp. strain B13, a 3-chlorobenzoate-metabolizing strain, were developed. Using this primer set and probe, we were able to detect Pseudomonas sp. strain B13 DNA sequences in DNA extracted from aquifer samples 14.5 months after Pseudomonas sp. strain B13 had been injected into a sand and gravel aquifer. This primer set and probe were also used to analyze isolates from 3-chlorobenzoate enrichments of the aquifer samples by Southern blot analysis. Hybridization of Southern blots with the Pseudomonas sp. strain B13-specific probe and a catabolic probe in conjunction with restriction fragment length polymorphism (RFLP) analysis of ribosome genes was used to determine that viable Pseudomonas sp. strain B13 persisted in this environment. We isolated a new 3-chlorobenzoate-degrading strain from one of these enrichment cultures. The B13-specific probe does not hybridize to DNA from this isolate. The new strain could be the result of gene exchange between Pseudomonas sp. strain B13 and an indigenous bacterium. This speculation is based on an RFLP pattern of ribosome genes that differs from that of Pseudomonas sp. strain B13, the fact that identically sized restriction fragments hybridized to the catabolic gene probe, and the absence of any enrichable 3-chlorobenzoate-degrading strains in the aquifer prior to inoculation.

Massachusetts↗

Degradation of trichloroethylene by Pseudomonas cepacia G4 and the constitutive mutant strain G4 5223 PR1 in aquifer microcosms

Pseudomonas cepacia G4 degrades trichloroethylene (TCE) via a degradation pathway for aromatic compounds which is induced by substrates such as phenol and tryptophan. P. cepacia G4 5223 PR1 (PR1) is a Tn5 insertion mutant which constitutively expresses the toluene ortho -monooxygenase responsible for TCE degradation. In groundwater microcosms, phenol-induced strain G4 and noninduced strain PR1 degraded TCE (20 and 50 μM) to nondetectable levels (< 0.1 μM) within 24 h at densities of 10 8 cells per ml; at lower densities, degradation of TCE was not observed after 48 h. In aquifer sediment microcosms, TCE was reduced from 60 to < 0.1 μM within 24 h at 5 x 10 8 PR1 organisms per g (wet weight) of sediment and from 60 to 26 μM over a period of 10 weeks at 5 x 10 7 PR1 organisms per g. Viable G4 and PR1 cells decreased from approximately 10 7 to 10 4 per g over the 10-week period.

Applied and Environmental Microbiology↗