Effects of antecedent fermentative and respiratory growth on the detection of chloramine-stressed Escherichia coil and Salmonella typhimurium.

In vitro laboratory studies were performed to assess the effects of antecedent growth conditions on the recovery of Escherichia coli ATCC 25922 and Salmonella typhimurium ATCC 14028 following chloramine disinfection. Six- and 18-h cultures of each organism were grown under aerobic, fermentative, and nitrate-reducing conditions prior to disinfection. At predetermined time intervals during a 10-min exposure to chloramine, survivors were surface plated on nonselective recovery media to determine C(n)t values. It was observed that nitrate-reducing growth predisposed the test organisms towards an increased sensitivity to chloramine stress over cells grown under fermentation or aerobic conditions (p < 0.01).

Evidence for Acquisition in Nature of a Chromosomal 2,4-Dichlorophenoxyacetic Acid/(alpha)-Ketoglutarate Dioxygenase Gene by Different Burkholderia spp.

We characterized the gene required to initiate the degradation of 2,4-dichlorophenoxyacetic acid (2,4-D) by the soil bacterium Burkholderia sp. strain TFD6, which hybridized to the tfdA gene of the canonical 2,4-D catabolic plasmid pJP4 under low-stringency conditions. Cleavage of the ether bond of 2,4-D by cell extracts of TFD6 proceeded by an (alpha)-ketoglutarate-dependent reaction, characteristic of TfdA (F. Fukumori and R. P. Hausinger, J. Bacteriol. 175:2083-2086, 1993). The TFD6 tfdA gene was identified in a recombinant plasmid which complemented a tfdA transposon mutant of TFD6 created by chromosomal insertion of Tn5. The plasmid also expressed TfdA activity in Escherichia coli DH5(alpha), as evidenced by enzyme assays with cell extracts. Sequence analysis of the tfdA gene and flanking regions from strain TFD6 showed 99.5% similarity to a tfdA gene cloned from the chromosome of a different Burkholderia species (strain RASC) isolated from a widely separated geographical area. This chromosomal gene has 77.2% sequence identity to tfdA from plasmid pJP4 (Y. Suwa, W. E. Holben, and L. J. Forney, abstr. Q-403, in Abstracts of the 94th General Meeting of the American Society for Microbiology 1994.). The tfdA homologs cloned from strains TFD6 and RASC are the first chromosomally encoded 2,4-D catabolic genes to be reported. The occurrence of highly similar tfdA genes in different bacterial species suggests that this chromosomal gene can be horizontally transferred.

Adaptation of Denitrifying Populations to Low Soil pH.

Natural denitrification rates and activities of denitrifying enzymes were measured in an agricultural soil which had a 20-year past history of low pH (pH ca. 4) due to fertilization with acid-generating ammonium salts. The soil adjacent to this site had been limed and had a pH of ca. 6.0. Natural denitrification rates of these areas were of similar magnitude: 158 ng of N g of soil day for the acid soil and 390 ng of N g of soil day at the neutral site. Estimates of in situ denitrifying enzyme activity were higher in the neutral soil, but substantial enzyme activity was also detected in the acid soil. Rates of nitrous oxide reduction were very low, even when NO(3) and NO(2) were undetectable, and were ca. 400 times lower than the rates of N(2)O production from NO(3). Denitrification rates measured in slurries of the acid and neutral soil showed distinctly different pH optima (pH 3.9 and pH 6.3) which were near the pH values of the two soils. This suggests that an acid-tolerant denitrifying population had been selected during the 20-year period of low pH.

Denitrification: ecological niches, competition and survival.

Organisms with the denitrification capacity are widely distributed and in high density in nature. It is not well understood why they are so successful. A survey of denitrifying enzyme content of various habitats is presented which indicates a role of carbon and oxygen, but not nitrate, in affecting denitrifier populations. It is suggested that organic carbon is more important than oxygen status in determining denitrifying enzyme content of habitats. In low oxygen environments, denitrifiers compete with organisms that dissimilate nitrate to ammonium, a process which conserves nitrogen. The energetic and kinetic parameters that affect this competition are evaluated. The latter is examined using Michaelis-Menten theoretical models by varying Vmax, Km, and So (substrate concentration) for the two competing populations. The outcome predicted by these models is presented and discussed in relation to previous data on population densities and Km values for representatives of these competing groups. These models suggest the conditions required to achieve changes in partitioning between the two fates of nitrate. These considerations are important if one is to be able to evaluate and successfully "manage" the fate of nitrate in any habitat.

Response of microbial populations in arctic tundra soils to crude oil.

Experimental crude oil spillages of 5 and 12 litre/m2 were established on the four major topographically distinguished soils of Arctic coastal polygonized tundra. The response of microbial populations to contaminating oil was found to depend on soil type and depth. Increases in numbers of heterotrophs were initially restricted to the top 2 cm of the soils. Increase in oil-degrading populations were found in oil-treated soils. Increases in microbial populations in subsurface soils paralleled downward migration of the oil. Some of the observed population increases probably resulted from input of plant residues and products from oil biodegradation.