Individual Microbial Species Research Articles

Digitization of DGGE (denaturing gradient gel electrophoresis) profile and cluster analysis of microbial communities

Denaturing gradient gel electrophoresis (DGGE) of 16S rDNA profiles were objectively digitized using an image analyzer; the individual microbial species in a community can thus be precisely quantified. The similarity between various microbial communities was compared to the digitized DGGE profiles using the cluster analyses technique. The microbial community in a biofilm was considerably different from that in suspended sludge obtained from the same system.

Depletion of pre-16S rRNA in starved Escherichia coli cells.

Specific hybridization assays for intermediates in rRNA synthesis (pre-rRNA) may become useful for monitoring the growth activity of individual microbial species in complex natural systems. This possibility depends upon the assumption that rRNA processing in microbial cells continues after growth and pre-rRNA synthesis cease, resulting in drainage of the pre-rRNA pool. This is not the case in many eukaryotic cells, but less is known about the situation in bacteria. Therefore, we used DNA probes to measure steady-state cellular pre-16S rRNA pools during growth state transitions in Escherichia coli. Pre-16S rRNA became undetectable when cells entered the stationary phase on rich medium and was replenished upon restoration of favorable growth conditions. These fluctuations were of much greater magnitude than concurrent fluctuations in the mature 16S rRNA pool. The extent of pre-16S rRNA depletion depended upon the circumstances limiting growth. It was significantly more pronounced in carbon-energy-starved cells than in nitrogen-starved cells or in cells treated with energy uncouplers. In the presence of the transcriptional inhibitor rifampin, rates of pre-16S rRNA depletion in carbon-energy-starved cells and nitrogen-starved cells were similar, suggesting that the difference between these conditions resides primarily at the level of pre-rRNA synthesis. Chloramphenicol, which inhibits the final steps in rRNA maturation, halted pre-16S rRNA depletion under all conditions. The data show that E. coli cells continue to process pre-rRNA after growth and rrn operon transcription cease, leading to drainage of the pre-rRNA pool. This supports the feasibility of using pre-rRNA-targeted probes to monitor bacterial growth in natural systems, with the caveat that patterns of pre-rRNA depletion vary with the conditions limiting growth.

Modified extended aeration process for removal and recovery of cadmium from wastewaters

It has been reported in the literature that several microbial species have a considerable sorption capacity for heavy metals. Work herein reported, accomplished using a heterogeneous microbial population developed in a modified extended aeration activated sludge process, indicates sorption capabilities as good as, or better than, those for individual microbial species. The process herein described may have several advantages over pure or controlled culture processes and it offers the possibility for reuse of organic matter needed to regenerate metal-sorbing biomass, as well as autodigestion of excess biomass. The work also indicates that acclimation to high levels of a specific heavy metal (Cd) does not confer acclimation to another heavy metal (Cu).

Sorption and Metabolism of Metolachlor by a Bacterial Community

A stable bacterial community absorbed and transformed the herbicide metolachlor [2-chloro-N-(2-ethyl- 6-methylphenyl)-N-(2-methoxy-1-methylethyl)-acetamide] from a liquid medium. About 80% of the added ring-[U-C]metolachlor (50 mug/ml) disappeared from the medium and accumulated inside the cells. The ratio of cellular C to C in 1 mg of supernatant reached a value of 1.1 x 10 in a 10-day-old culture. C remaining in the medium consisted primarily of two dechlorinated products of metolachlor with m/z 233 and 263 as determined by mass spectrometry. The C-labeled material absorbed by the cells was strongly bound; only 2% of the C was released into deionized water after shaking for 3 h. Approximately 96% of the C associated with the biomass was extracted with acetone, and high-performance liquid chromatographic analysis of this fraction showed six peaks containing radioactivity. Since no metolachlor was detected by chromatographic analysis, it was concluded that the radioactivity recovered from the cells represented transformed products of metolachlor. Pure cultures isolated from the bacterial mixed culture were less effective in transforming and accumulating metolachlor. These results suggest that it may be advantageous to seed an aquatic environment with a mixture of microorganisms, rather than individual microbial species, as a method for removal or detoxification of metolachlor.

Nature and significance of microbial cometabolism of xenobiotics.

Microbial cometabolism, i.e. "transformation of a non-growth substrate in the obligate presence of a growth substrate or another transformable compound" (Dalton and Stirling 1982) is a whole-cell phenomenon physiologically based on coupling of different catabolic pathways at the cellular level. It is frequently observed in transformation of xenobiotic non-growth substrates by individual microbial species. Transformation processes of this type are usually mediated by appropriate non-specific enzymes of the peripheric cellular metabolism able to modify a variety of substances other than their natural substrates. The precise mechanisms of coupling between metabolism of xenobiotic non-growth substrates and of particular additional carbon substrates may be different depending on the substrates and the microbial species involved. However, experimental data indicate that the primary function of the respective additional carbon substrates is to supply either energy, cofactors or metabolites for the different cellular events involved in the transformation process (e.g. uptake of the xenobiotic non-growth substrate, functioning of appropriate degradative enzymes of the peripheric cellular metabolism). Cometabolism of xenobiotics involves nothing special or novel from the standpoint of biochemistry. On the contrary, there are numerous examples where the turnover of particular natural compounds by certain aerobic or anaerobic microorganisms is essentially based on coupling of different catabolic pathways at the cellular level by transfer of hydrogen (i.e. reducing power) and/or energy between two or more enzymatic reactions. Synthetic chemicals which resist total degradation by individual microbial species may undergo mineralization due to complementary catabolic sequences mediated by certain multispecies microbial associations with cometabolic transformations being the initial steps. Although taking place in certain natural habitats (e.g. rhizospheres, sewage), microbial cometabolism of xenobiotics in natural ecosystems occurs with slow rates since the respective cometabolizing populations are generally small and will not increase in number or biomass in response to the introduced chemicals. However, under conditions of axenic microbial cultures, high concentrations of biomass, and appropriate substrate mixtures cometabolism of synthetic chemicals may be a useful technique of considerable practical importance to accumulate biochemical products at high yields. In addition, cometabolic capabilities of wild-type microorganisms may serve as a tool for the construction of microbial strains with a new degradative potential for recalcitrant xenobiotic compounds.

The bacteriology and protozoology of ruminant digestion

AbstractRecent successes in the pure culture study of true rumen bacteria responsible for carbohydrate fermentations are outlined. Some account is also given of the rumen ciliate protozoa as fermentative agents. The power of polysaccharide synthesis and storage is very marked in many of these biochemically active rumen micro‐organisms, both bacteria and protozoa. On the other hand little is yet known about individual microbial species responsible for extensive nitrogenous transformations in the rumen.