Effect of temperature on mineralization by heterotrophic bacteria.

When pure cultures of the bacteria Pseudomonas fluorescens (a psychrotroph), Escherichia coli (a mesophile), and SRL 261 (a thermophile) were shifted away from temperatures to which they were adapted, the percentage of substrate mineralized increased (percent mineralized = [substrate respired to CO2]/substrate respired to CO2 + substrate incorporated into biomass] X 100). The increase in the percent mineralized was larger for larger temperature shifts. Similar responses were observed when natural heterotrophic bacterial populations from sediments of Lake George, N.Y., and a thermophilic algal-bacterial mat community at the Savannah River Plant, Aiken, S.C., were subjected to temperature shifts. These results suggest that an increase in the percent mineralized may be an indication of thermal stress in bacterial populations.     

Chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria.

Comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. A bacterial consortium, designated LPS10, was shown to mineralize 4-chlorobiphenyl (4CB) and dehalogenate 4,4'-dichlorobiphenyl. The LPS10 consortium involved three isolates: Pseudomonas testosteroni (LPS10A), which mediated the breakdown of 4CB and 4,4'-dichlorobiphenyl to 4-chlorobenzoic acid; an isolate tentatively identified as an Arthrobacter sp. (LPS10B), which mediated 4-chlorobenzoic acid degradation; and Pseudomonas putida bv. A (LPS10C), whose role in the consortium has not been determined. None of these isolates contained detectable plasmids or sequences homologous to the 4CB-degradative plasmid pSS50. A freshwater isolate, designated LBS1C1, was found to harbor a 41-megadalton plasmid that was related to the 35-megadalton plasmid pSS50, and this isolate was shown to mineralize 4CB. In chemostat enrichments with biphenyl and 4CB as primary carbon sources, the LPS10 consortium was found to outcomplete bacterial populations harboring plasmids homologous to pSS50. These results demonstrate that an understanding of the biodegradative capacity of individual bacterial populations as well as interacting populations of bacteria must be considered in order to gain a better understanding of polychlorinated biphenyl biodegradation in the environment.     

Role of heterotrophic bacteria in complete mineralization of trichloroethylene by Methylocystis sp. strain M.

Biodegradation experiments with radioactively labeled trichloroethylene showed that 32% of the radioactive carbon was converted to glyoxylic acid, dichloroacetic acid and trichloroacetic acid and that the same percentage was converted to CO2 and CO after 140 h of incubation by a pure culture of a type II methane-utilizing bacterium, Methylocystis sp. strain M, isolated from a mixed culture, MU-81, in our laboratory. In contrast, these water-soluble (14C)trichloroethylene degradation products were completely or partially degraded further and converted to CO2 by the MU-81 mixed culture. This phenomenon was attributed to the presence of a heterotrophic bacterium (strain DA4), which was identified as Xanthobacter autotrophicus, in the MU-81 culture. The results indicate that the heterotrophic bacteria play an important role in complete trichloroethylene degradation by methanotrophs.     

Chloroform aerobic cometabolism by butane-utilizing bacteria in bioaugmented and non-bioaugmented soil/groundwater microcosms

The aerobic cometabolic chloroform (CF) degradation by butane-growing biomasses was investigated in slurry microcosms. The lag-time for the onset of butane utilization by the indigenous biomass of the studied sandy soil was less than 2 weeks in all the experimental conditions tested. The shortest lags were obtained in the absence of CF. The lag-time for the onset of CF depletion was strongly affected by temperature, with no CF degradation after several weeks in the tests conducted at 15 °C. Bioaugmentation treatments performed with two types of butane-utilizing inocula led to a marked decrease of the butane lag-time, even at the smallest concentration of augmented bacteria tested (3.5 × 10³ CFU/mL_aq. phase). Tests of prolonged CF degradation in the absence of butane were satisfactorily simulated with a Monod-type kinetic model. Estimates of the minimum butane/CF molar ratio required to sustain CF cometabolism varied from 2.0 to 3.1.     

Role of heterotrophic bacteria in complete mineralization of trichloroethylene by Methylocystis sp. strain M.

Biodegradation experiments with radioactively labeled trichloroethylene showed that 32% of the radioactive carbon was converted to glyoxylic acid, dichloroacetic acid and trichloroacetic acid and that the same percentage was converted to CO2 and CO after 140 h of incubation by a pure culture of a type II methane-utilizing bacterium, Methylocystis sp. strain M, isolated from a mixed culture, MU-81, in our laboratory. In contrast, these water-soluble (14C)trichloroethylene degradation products were completely or partially degraded further and converted to CO2 by the MU-81 mixed culture. This phenomenon was attributed to the presence of a heterotrophic bacterium (strain DA4), which was identified as Xanthobacter autotrophicus, in the MU-81 culture. The results indicate that the heterotrophic bacteria play an important role in complete trichloroethylene degradation by methanotrophs.     

Chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria

Comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. A bacterial consortium, designated LPS10, was shown to mineralize 4-chlorobiphenyl (4CB) and dehalogenate 4,4'-dichlorobiphenyl. The LPS10 consortium involved three isolates: Pseudomonas testosteroni (LPS10A), which mediated the breakdown of 4CB and 4,4'-dichlorobiphenyl to 4-chlorobenzoic acid; an isolate tentatively identified as an Arthrobacter sp. (LPS10B), which mediated 4-chlorobenzoic acid degradation; and Pseudomonas putida bv. A (LPS10C), whose role in the consortium has not been determined. None of these isolates contained detectable plasmids or sequences homologous to the 4CB-degradative plasmid pSS50. A freshwater isolate, designated LBS1C1, was found to harbor a 41-megadalton plasmid that was related to the 35-megadalton plasmid pSS50, and this isolate was shown to mineralize 4CB. In chemostat enrichments with biphenyl and 4CB as primary carbon sources, the LPS10 consortium was found to outcomplete bacterial populations harboring plasmids homologous to pSS50. These results demonstrate that an understanding of the biodegradative capacity of individual bacterial populations as well as interacting populations of bacteria must be considered in order to gain a better understanding of polychlorinated biphenyl biodegradation in the environment.     

Degradation and mineralization of petroleum by two bacteria isolated from coastal waters

Within the framework of a study on the oil biodegradation potential of the sea the ability of a Flavobacterium sp. and Brevibacterium sp. to metabolize a paraffinic crude oil and a chemically defined hydrocarbon mixture was investigated. Major components of the crude oil were identified by combination gas chromatography and mass spectrometry. The rate and extent of total hydrocarbon biodegradation was measured. In addition, CO₂ evolution from the crude oil was continuously monitored in a shaker-mounted gas train arrangement. Degradation started after a 2 to 4 day lag period, and reached its maximum within two weeks. At this time up to 60% of the crude oil and 75% of the model hydrocarbon mixture, each added at the level of 1 ml per 100 ml artificial sea water, were degraded. Mineralization(conversion to CO₂) was slightly lower due to formation of products and bacterial cell material. n-Paraffins were preferentially degraded as compared to branched chain hydrocarbons. Biodegradation of n-paraffins in the range of C₁₂ to C₂₀ was simultaneous; no diauxie effects were observed.     

Enhanced mineralization of polychlorinated biphenyls in soil inoculated with chlorobenzoate-degrading bacteria.

An Altamont soil containing no measurable population of chlorobenzoate utilizers was examined for the potential to enhance polychlorinated biphenyl (PCB) mineralization by inoculation with chlorobenzoate utilizers, a biphenyl utilizer, combinations of the two physiological types, and chlorobiphenyl-mineralizing transconjugants. Biphenyl was added to all soils, and biodegradation of 14C-Aroclor 1242 was assessed by disappearance of that substance and by production of 14CO2. Mineralization of PCBs was consistently greatest (up to 25.5%) in soils inoculated with chlorobenzoate degraders alone. Mineralization was significantly lower in soils receiving all other treatments: PCB cometabolizer (10.7%); chlorobiphenyl mineralizers (8.7 and 14.9%); and mixed inocula of PCB cometabolizers and chlorobenzoate utilizers (11.4 and 18.0%). However, all inoculated soils had higher mineralization than did the uninoculated control (3.1%). PCB disappearance followed trends similar to that observed with the mineralization data, with the greatest degradation occurring in soils inoculated with the chlorobenzoate-degrading strains Pseudomonas aeruginosa JB2 and Pseudomonas putida P111 alone. While the mechanism by which the introduction of chlorobenzoate degraders alone enhanced biodegradation of PCBs could not be elucidated, the possibility that chlorobenzoate inoculants acquired the ability to metabolize biphenyl and possibly PCBs was explored. When strain JB2, which does not utilize biphenyl, was inoculated into soil containing biphenyl and Aroclor 1242, the frequency of isolates able to utilize biphenyl and 2,5-dichlorobenzoate increased progressively with time from 3.3 to 44.4% between 15 and 48 days, respectively. Since this soil contained no measurable level of chlorobenzoate utilizers yet did contain a population of biphenyl utilizers, the possibility of genetic transfer between the latter group and strain JB2 cannot be excluded.     

Tree species mixture inhibits soil organic carbon mineralization accompanied by decreased r-selected bacteria

Background and aims

Fine-root functioning is a major driver of plant growth and strongly influences the global carbon cycle. While fine-root over-yielding has been shown in the upper soil layers of mixed-species forests relative to monospecific stands, the consequences of tree diversity on fine-root growth in very deep soil layers is still unknown. Our study aimed to assess the consequences of mixing Acacia mangium and Eucalyptus grandis trees on soil exploration by roots down to the water table at 17 m depth in a tropical planted forest.

Method

Fine roots (diameter < 2 mm) were sampled in a randomized block design with three treatments: monospecific stands of Acacia mangium (100A), Eucalyptus grandis (100E), and mixed stands with 50% of each species (50A50E). Root ingrowth bags were installed at 4 depths (from 0.1 m to 6 m) in the three treatments within three different blocks, to study the fine-root production over 2 periods of 3 months.

Results

Down to 17 m depth, total fine-root biomass was 1127 g m^?2 in 50A50E, 780 g m^?2 in 100A and 714 g m^?2 in 100E. Specific root length and specific root area were 110–150% higher in 50A50E than in 100A for Acacia mangium trees and 34% higher in 50A50E than in 100E for Eucalyptus grandis trees. Ingrowth bags showed that the capacity of fine roots to explore soil patches did not decrease down to a depth of 6 m for the two species.

Conclusions

Belowground interactions between Acacia mangium and Eucalyptus grandis trees greatly increased the exploration of very deep soil layers by fine roots, which is likely to enhance the uptake of soil resources. Mixing tree species might therefore increase the resilience of tropical planted forests through a better exploration of deep soils.

     

Dechlorination of Chloroform by Methanosarcina Strains

Dehalogenation of carbon tetrachloride, chloroform, and bromoform in pure cultures of Methanosarcina sp. strain DCM and Methanosarcina mazei S6 was demonstrated. The initial dechlorination product of chloroform was methylene chloride (dichloromethane), which accumulated transiently to about 70% of the added chloroform; trace amounts of chloromethane were also detected. The amount of chloroform dechlorinated per mole of methane produced was approximately 10 times greater than the ratio observed previously for tetrachloroethene dechlorination by these strains. The production of ¹⁴CO₂ from [¹⁴C]chloroform and the absence of ¹⁴CH₄ imply that processes in addition to reductive dechlorination operate.     

Anaerobic mineralization of quaternary carbon atoms: isolation of denitrifying bacteria on dimethylmalonate.

The microbial capacity to degrade simple organic compounds with quaternary carbon atoms was demonstrated by enrichment and isolation of five denitrifying strains on dimethylmalonate as the sole electron donor and carbon source. Quantitative growth experiments showed a complete mineralization of dimethylmalonate. According to phylogenetic analysis of the complete 16S rRNA genes, two strains isolated from activated sewage sludge were related to the genus Paracoccus within the alpha-Proteobacteria (98.0 and 98.2% 16S rRNA gene similarity to Paracoccus denitrificans(T)), and three strains isolated from freshwater ditches were affiliated with the beta-Proteobacteria (97.4 and 98.3% 16S rRNA gene similarity to Herbaspirillum seropedicae(T) and Acidovorax facilis(T), respectively). Most-probable-number determinations for denitrifying populations in sewage sludge yielded 4.6 x 10(4) dimethylmalonate-utilizing cells ml(-1), representing up to 0.4% of the total culturable nitrate-reducing population.     

Phenanthrene mineralization by Pseudomonas sp. UG14

A phenanthrene-mineralizing Pseudomonas sp., designated UG14, was isolated from creosote-contaminated soil. It contained two plasmids, of approximately 77 kb and 76 kb, the smaller of which contained DNA sequences that hybridized with probes specific for ndoB and xylE, genes involved in catabolism of aromatic hydrocarbons. At initial phenanthrene concentrations of 10, 50, 200 and 1000 mg/l broth, 27%, 19%, 7.7% and 3.3%, respectively, of the [9-¹⁴C]phenanthrene was recovered as ¹⁴CO₂ after 36 days' incubation at 30°C. Most ¹⁴C-label was converted to a water-soluble metabolite tentatively identified as 1-hydroxy-2-naphthoic acid. Rhamnolipid biosurfactants produced by P. aeruginosa UG2 enhanced mineralization of 50 mg phenanthrene/l by Pseudomonas sp. UG14. With the biosurfactant at 0, 25 and 250 mg rhamnose equivalents/l, 6.5%, 8.2% and 9.8%, respectively, of the phenanthrene was mineralized after 35 days.M.A. Providenti, H. Lee and J.T. Trevors are with the Department of Environmental Biology, University of Guelph, Guelph, Ontario, N1G 2W1, Canada; C.W. Greer is with the National Research Council Canada, Biotechnology Research Institute, 6100 Royalmount Ave, Montreal, Quebec, H4P 2R2, Canada.