Entire Microbial Community Research Articles

Pico- and nanoplankton dynamics during bloom initiation of Aureococcus in a Long Island, NY bay

The entire microbial plankton community was quantified on a weekly basis April through June of 2000 in Quantuck Bay as part of an ongoing study to identify factors contributing to the initiation of blooms of Aureococcus anophagefferens (brown tide) in Long Island, NY bays. We used flow cytometry, imaging cytometry, fluorescent antibody cell counts, and traditional visual cell counting to quantify the picophytoplankton, heterotrophic bacteria, nanophytoplankton, heterotrophic protists, and microplankton prior to, and during the initiation of a brown tide bloom. Cells passing through a 5 μm mesh dominated the total chlorophyll concentration (>80%) for most of the spring study period. The A. anophagefferens bloom occurred in the context of a larger pico/nanophytoplankton bloom where A. anophagefferens accounted for only 30% of the total cell count when it was at its maximum concentration of 4.8 × 10 5 mL −1. Levels of dissolved organic nitrogen were enriched during the bloom peak relative to pre-bloom levels and heterotrophic bacteria also bloomed, reaching abundances over 10 7 mL −1. A trophic cascade within the heterotrophic protist community may have occurred, coinciding with the A. anophagefferens bloom. Before the onset of the bloom, larger grazers increased in abundance, while the next smaller trophic level of grazers were diminished. These smaller grazers were the likely water column predators of A. anophagefferens, and the brown tide bloom initiated when they were depleted. These results suggest that this bloom initiated due to interactions with other pico/nano algae and release from grazing pressure through a trophic cascade.

Effect of anoxia and high sulphide concentrations on heterotrophic microbial communities in reduced surface sediments (Black Spots) in sandy intertidal flats of the German Wadden Sea

Black reduced sediment surfaces (Black Spots) in sandy intertidal flats of the German Wadden Sea (southern North Sea) are characterised by elevated sulphide concentrations (up to 20 mM) and low redox potentials. It is assumed that the appearance of Black Spots is linked to elevated levels of organic matter content within the sediments. In order to establish the effect of high substrate and sulphide concentrations on the heterotrophic microbial communities in Black Spot sediments, bacterial abundances and the potential C-source utilisation patterns of microbial communities were compared in natural and artificially induced Black Spots and unaffected control sites. Bacterial numbers were estimated by direct counts and the most probable number technique for different physiological groups, while patterns of C-substrate utilisation of entire aerobic microbial communities were assessed using the Biolog™ sole-carbon-source-catabolism assay. Bacterial abundances at Black Spot sites were increased, with increases in mean cell numbers, more disperse data distributions and more extreme values. Substrate utilisation patterns of aerobic microbial communities were significantly different in Black Spot sediment slurries, showing diminished richness (number of C-sources catabolised) and substrate diversity (Shannon diversity index) in comparison to unaffected sites. Principal component analysis clearly discriminated Black Spot utilisation patterns from controls and indicated that microbial communities in individual Black Spot sites are functionally diverse and differ from communities in oxidised surface sediments and reduced subsurface sediments at control sites. This work suggests that potentially negative effects on microbial communities in Black Spot sediments, through anoxia and high sulphide concentrations, are balanced by the stimulating influence of substrate availability, leading to comparable or higher bacterial numbers, but lower functional microbial diversity of aerobic microbial communities.

Culture-independent microbial community analysis reveals that inulin in the diet primarily affects previously unknown bacteria in the mouse cecum.

Inulin is a well-known fructose-based prebiotic which has been shown to stimulate the growth of bifidobacteria, a bacterial group generally considered beneficial for intestinal health. In the present study, we analyzed inulin-associated shifts in the total bacterial community of wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene by using DNA-based approaches independent of bacterial culturability. Mice were fed a high-fat, nonfiber diet with or without inulin inclusion at a 10% (wt/wt) concentration. Cecal contents were analyzed after 0, 3, and 9 weeks on the experimental diets. Inulin inclusion significantly affected the total bacterial community structure of the cecum as determined by both a nonselective percent-guanine-plus-cytosine-based profiling analysis and a more specific 16S ribosomal DNA sequence analysis. The shifts included stimulation of bifidobacteria and suppression of clostridia, but sequence comparison revealed that the major shifts were within previously unknown bacterial taxa. Concomitantly, significantly higher bacterial densities, determined by flow cytometry, were observed with the inulin-amended diet, and the metabolism of the cecal bacterial community was altered, as indicated by higher levels of residual short-chain fatty acids, particularly lactic acid. With regard to all of the microbiological parameters measured, the wild-type mice and mice carrying a genetically inactivated adenomatous polyposis coli tumor suppressor gene were essentially identical. Studies of the implications of pre- and probiotics may need to be expanded to include careful analysis of their effects on the entire microbial community, rather than just a few well-known species. Further studies are needed to increase our understanding of the possible roles of currently unknown gastrointestinal bacteria in health and disease.

Bacterial growth and grazing on diatom aggregates: Respiratory carbon turnover as a function of aggregate size and sinking velocity

Bacterial growth, respiration, particulate organic carbon (POC), and particulate organic nitrogen (PON) were measured directly on differentially sized diatom aggregates incubated individually in suspension in order to study the coupling between these parameters under controlled conditions. After 3 d ofincubation, bacteria, flagellates, and ciliates were present on aggregates in the ratio of 1,100:30:1. Bacterial generation times ranged from 0.4 to 2 d, but these short generation times did not result in an increase of bacterial abundance because bacteria were grazed approximately at similar rates. The entire microbial community respired 2.90 carbon units for each carbon unit incorporated by the bacteria. Bacterial production, community respiration, POC, and PON increased with increasing aggregate size, and respiration was proportional to POC and PON content. The POC specific respiration rate on aggregates was 0.083 d−1, and 40% of the initial POC content was respired after 6 d. From simple calculations combining carbon‐specific respiration rates and aggregate sinking velocities, it is concluded that a tight coupling between POC and microbial respiration may control the carbon fluxes of sinking diatom aggregates in the ocean.

Molecular Techniques for Determining Microbial Diversity and Community Structure in Natural Environments

The ability to quantify the number and kinds of microorganisms within a community is fundamental to the understanding of the structure and function of an ecosystem. The simple morphology of most microbes provides few clues for their identification and physiological traits are often ambiguous. In addition, many organisms resist cultivation, which is essential to their characterization. Recombinant DNA techniques have provided a means whereby many of the obstacles associated with cultivation and description can be overcome and subsequently has allowed many new insights into the complexity of natural microbial communities. Molecular approaches based on 16S ribosomal RNA (rRNA) sequence analysis allow direct investigation of the community structure, diversity, and phylogeny of microorganisms in almost any environment, while quantification of the individual types of microorganisms or entire microbial communities may be addressed by nucleic acid hybridization techniques. Furthermore, the use of fluorescently labeled population-specific rRNA probes allows microscopic examination of individual cells in complex microbial assemblages as well as their interactions in situ. In this review, we discuss strategies for characterizing microbial communities without the need for cultivation.

Microbial Diversity and Community Structure in Two Different Agricultural Soil Communities.

Abstract In this study, two different agricultural soils were investigated: one organic soil and one sandy soil, from Stend (south of Bergen), Norway. The sandy soil was a field frequently tilled and subjected to crop rotations. The organic soil was permanent grazing land, infrequently tilled. Our objective was to compare the diversity of the cultivable bacteria with the diversity of the total bacterial population in soil. About 200 bacteria, randomly isolated by standard procedures, were investigated. The diversity of the cultivable bacteria was described at phenotypic, phylogenetic, and genetic levels by applying phenotypical testing (Biolog) and molecular methods, such as amplified rDNA restriction analysis (ARDRA); hybridization to oligonucleotide probes; and REP-PCR. The total bacterial diversity was determined by reassociation analysis of DNA isolated from the bacterial fraction of environmental samples, combined with ARDRA and DGGE analysis. The relationship between the diversity of cultivated bacteria and the total bacteria was elucidated. Organic soil exhibited a higher diversity for all analyses performed than the sandy soil. Analysis of cultivable bacteria resulted in different resolution levels and revealed a high biodiversity within the population of cultured isolates. The difference between the two agricultural soils was significantly higher when the total bacterial population was analyzed than when the cultivable population was. Thus, analysis of microbial diversity must ultimately embrace the entire microbial community DNA, rather than DNA from cultivable bacteria.

Ecofunctional enzymes of microbial communities in ground water.

Biolog technology was initially developed as a rapid, broad spectrum method for the biochemical identification of clinical microorganisms. Demand and creative application of this technology has resulted in the development of Biolog plates for Gram-negative and Gram-positive bacteria, for yeast and Lactobacillus sp. Microbial ecologists have extended the use of these plates from the identification of pure culture isolates to a tool for quantifying the metabolic patterns of mixed cultures, consortia and entire microbial communities. Patterns that develop on Biolog microplates are a result of the oxidation of the substrates by microorganisms in the inoculum and the subsequent reduction of the tetrazolium dye to form a color in response to detectable reactions. Depending upon the functional enzymes present in the isolate or community one of a possible 4 x 10(28) patterns can be expressed. The patterns were used to distinguish the physiological ecology of various microbial communities present in remediated groundwater. The data indicate that one can observe differences in the microbial community among treatments of bioventing, 1% and 4% methane injection, and pulse injection of air, methane and nutrients both between and among wells. The investigation indicates that Biolog technology is a useful parameter to measure the physiological response of the microbial community to perturbation and allows one to design enhancement techniques to further the degradation of selected recalcitrant and toxic chemicals. Further it allows one to evaluate the recovery of the microbial subsurface ecosystem after the perturbations have ceased. We propose the term 'ecofunctional enzymes' (EFE) as the most descriptive and useful term for the Biolog plate patterns generated by microbial communities. We offer this designation and provide ecological application in an attempt to standardize the terminology for this relatively new and unique technology.

Characterization of marine prokaryotic communities via DNA and RNA

We know very little about species distributions in prokaryotic marine plankton. Such information is very interesting in its own right, and ignorance of it is also beginning to hamper process studies, such as those on viral infection. New DNA- and RNA-based approaches avoid many prior limitations. Here we discuss four such applications: (1) cloning and sequencing of 16S rRNA genes to produce lists of what types of organisms are present; (2) quantification of these individual types in marine samples by nucleic acid hybridization, including single cell fluorescence; (3) quantitative comparison by DNA-DNA hybridization of entire microbial communities in terms of shared common types, without knowledge of community components; and (4) finding cultures that are representative of native communities. Several previously uncharacterized types of bacteria and archaea (probably including novel phyla) are present in marine plankton. Evidence from both the Atlantic and Pacific suggests that as-of-yet uncultivated archaea may dominate the deep sea, and thus may be the most abundant group of organisms on Earth. Such archaea are in surface waters as well, and can be visualized with fluorescent probes and enriched at room temperature with addition of organic nutrients. Community hybridization shows that variability of microbial community compositions in time and space is high. Although most native bacteria do not grow in culture, some proteobacterial cultures appear by genomic hybridization to be representative of certain communities. These and other results indicate the utility of DNA- and RNA-based methods.

A method for measuring the response of sediment microbial communities to environmental perturbations

A method has been developed for studying direct and indirect responses of microbial processes in lake sediments to environmental perturbations. Responses of the entire microbial community as well as specific members of the community were studied using in vitro sediment systems for periods of weeks or months under control and experimentally perturbed conditions. Effects of perturbations at the community level were determined by comparing rates of organic matter decomposition (CO2 + CH4 production) in the control and test (acidified) systems. At the same time and under the same conditions, rates of specific processes such as mercury methylation and sulfate and nitrate reduction were assayed to see if these processes responded in the same manner as did the activity of the entire community. Acidification (lowering from pH 6.3 to 4.2) did not affect either community activity or nitrate reduction. However, decreases were observed in sulfate reduction and mercury methylation which were independent of community activity, suggesting that acidification may affect these two specific processes directly. Use of this method provides comprehensive information about the interaction of sediment microbial processes as they respond to environmental perturbations.