Patterns Of Microbial Diversity Research Articles

It's all relative: ranking the diversity of aquatic bacterial communities

The study of microbial diversity patterns is hampered by the enormous diversity of microbial communities and the lack of resources to sample them exhaustively. For many questions about richness and evenness, however, one only needs to know the relative order of diversity among samples rather than total diversity. We used 16S libraries from the Global Ocean Survey to investigate the ability of 10 diversity statistics (including rarefaction, non-parametric, parametric, curve extrapolation and diversity indices) to assess the relative diversity of six aquatic bacterial communities. Overall, we found that the statistics yielded remarkably similar rankings of the samples for a given sequence similarity cut-off. This correspondence, despite the different underlying assumptions of the statistics, suggests that diversity statistics are a useful tool for ranking samples of microbial diversity. In addition, sequence similarity cut-off influenced the diversity ranking of the samples, demonstrating that diversity statistics can also be used to detect differences in phylogenetic structure among microbial communities. Finally, a subsampling analysis suggests that further sequencing from these particular clone libraries would not have substantially changed the richness rankings of the samples.

Increasing ecological inference from high throughput sequencing of fungi in the environment through a tagging approach

High throughput sequencing methods are widely used in analyses of microbial diversity, but are generally applied to small numbers of samples, which precludes characterization of patterns of microbial diversity across space and time. We have designed a primer-tagging approach that allows pooling and subsequent sorting of numerous samples, which is directed to amplification of a region spanning the nuclear ribosomal internal transcribed spacers and partial large subunit from fungi in environmental samples. To test the method for phylogenetic biases, we constructed a controlled mixture of four taxa representing the Chytridiomycota, Zygomycota, Ascomycota and Basidiomycota. Following cloning and colony restriction fragment length polymorphism analysis, we found no significant difference in representation in 19 of the 23 tested primers. We also generated a clone library from two soil DNA extracts using two primers for each extract and compared 456 clone sequences. Community diversity statistics and contingency table tests applied to counts of operational taxonomic units revealed that the two DNA extracts differed significantly, while the pairs of tagged primers from each extract were indistinguishable. Similar results were obtained using UniFrac phylogenetic comparisons. Together, these results suggest that the pig-tagged primers can be used to increase ecological inference in high throughput sequencing projects on fungi.

Endolithic Microbial Ecosystems

The endolithic environment, the pore space in rocks, is a ubiquitous microbial habitat and an interface between biology and geology. Photosynthesis-based endolithic communities inhabit the outer centimeters of rocks exposed to the surface, and offer model systems for microbial ecology, geobiology, and astrobiology. Endolithic ecosystems are among the simplest microbial ecosystems known and as such provide tractable models for testing ecological hypotheses. Such hypotheses have been difficult to test because microbial ecosystems are extraordinarily diverse. We review here recent culture-independent, ribosomal RNA-based studies that evaluate hypotheses about endolithic ecosystems, and provide insight for understanding general principles in microbial ecology. Comparison of endolithic communities supports the principle that patterns of microbial diversity are governed by similar principles observed in macroecological systems. Recent results also explore geobiological processes that shape the current biosphere and potentially provide clues to life's history on Earth and where to seek life elsewhere in the Solar System.

The Modulating Role of Dissolved Organic Matter on Spatial Patterns of Microbial Metabolism in Lake Erie Sediments

To evaluate the role of dissolved organic matter (DOM) on microbial community metabolism, we established extracellular enzyme activity (EEA) and substrate-induced respiration (SIR) profiles of sediment samples collected from littoral and profundal regions of the western, central, and eastern basins of Lake Erie. Lake Erie is spatially structured such that the central and western basins receive relatively major inputs of allochthonous DOM in comparison to the eastern basin. Overall, spatial patterns of EEA and SIR profiles suggest both greater metabolic diversity and activity in the littoral regions of the central and western basins. In contrast, the eastern basin demonstrated much less structuring between littoral and profundal areas. To evaluate whether the observed spatial patterns are the result of microbial community adaptations to local DOM availability, we performed three experimental treatments by inoculating sediment samples with polyvinylpyrrolidone, which sequesters large polyphenols, or with either vanillin or catechol, two small phenolic compounds. Our results revealed that esterase and glycosidase EEA from the eastern basin were induced by small phenolics and inhibited by large polyphenols. In contrast, the addition of small phenolics decreased esterase and glycosidase activities from the central basin, while polyphenols had a negligible effect. These results suggest that the source and composition of DOM play a significant role in the local adaptation of microbial communities, determining large-scale spatial patterns of microbial functional diversity in Lake Erie sediments.

The cyanobacterial community of polygon soils at an inland Antarctic nunatak

Inland Antarctic terrestrial ecosystems and biodiversity are poorly understood in comparison with Antarctic coastal regions. Microorganisms, as primary colonists, are integral to Antarctic soil ecosystem development, essential for pedogenesis and structuring the soil, and providing the nutrients necessary for the subsequent establishment of macroorganisms. This study analysed the microbial communities present in polygon soils of Coal Nunatak (Alexander Island, at the southern limit of the maritime Antarctic). Soils were analysed across three polygons (centre and margins) and at three depths (0–1, 1–2, 2–5 cm). Cyanobacterial communities were characterised using two complementary molecular biological approaches, temperature gradient gel electrophoresis and clone library analysis. The three polygons exhibited conspicuous differences in community composition, both between different polygons and spatially (horizontally and vertically) within a single polygon. Comparison of our data with that from previous studies using classical culture and morphological identification techniques clearly shows the need for more intensive research on patterns of microbial diversity in terrestrial habitats throughout the Antarctic. The majority of the 17 cyanobacterial genera identified at Coal Nunatak are thought to have ubiquitous distributions, while none are known only from the Antarctic. Three of the genera present are also known to be capable of being lichen photobionts.

Terminal restriction fragment length polymorphism analysis of the diversity of fecal microbiota in patients with ulcerative colitis

Terminal restriction fragment length polymorphism (T-RFLP) analysis is a powerful tool to assess the diversity of complexed microbiota. This permits rapid comparison of microbiota from many samples. In this study, we performed T-RFLP analysis of the fecal microbiota from patients with ulcerative colitis (UC). Forty-four patients with UC (23 women and 21 men, median age 25 years) and 46 healthy individuals (25 women and 21 men, median age 34 years) were enrolled in this study. DNA was extracted from their stool samples, and the 16S rRNA genes were amplified by PCR. The PCR products were then digested with HhaI and/or MspI restriction enzymes, and the length of the T-RF was determined. The fecal microbial communities were classified in 8 clusters. Almost all the healthy individuals (39 of 46) were included in cluster I, and most of the UC patients could be divided into the other 7 clusters, indicating that fecal bacterial communities are different between healthy individuals and active UC patients. Some T-RFs, derived from the unclassified bacteria, Ruminococcus, Eubacterium, Fusobacterium, gammaproteobacteria, unclassified Bacteroides, and unclassified Lactobacillus, were detected in the UC patients, but not in the healthy individuals. The T-RFLP patterns were also different between the active patients and inactive (remission) patients. The T-RF derived from the unclassified bacteria, Ruminococcus and Eubacterium, and the T-RFs derived from the unclassified bacteria, Eubacterium, and Fusobacterium were predominantly detected in the active patients not the inactive patients. In contrast, the T-RFs derived from Lactobacillus and unclassified Lactobacillus were more predominant in the inactive (remission) patients. In 4 patients with proctitis, the pattern of fecal microbial diversity was very similar. T-RFLP analyses showed that the diversity of fecal microbiota in patients with UC was different from that in healthy individuals. Unclassified bacteria, as well as known bacteria, can contribute to alterations in the bacterial diversity of UC patients.

Phylogenetic Composition of Rocky Mountain Endolithic Microbial Ecosystems

The endolithic environment, the pore space in rocks, is a ubiquitous microbial habitat. Photosynthesis-based endolithic communities inhabit the outer few millimeters to centimeters of rocks exposed to the surface. Such endolithic ecosystems have been proposed as simple, tractable models for understanding basic principles in microbial ecology. In order to test previously conceived hypotheses about endolithic ecosystems, we studied selected endolithic communities in the Rocky Mountain region of the United States with culture-independent molecular methods. Community compositions were determined by determining rRNA gene sequence contents, and communities were compared using statistical phylogenetic methods. The results indicate that endolithic ecosystems are seeded from a select, global metacommunity and form true ecological communities that are among the simplest microbial ecosystems known. Statistical analysis showed that biogeographical characteristics that control community composition, such as rock type, are more complex than predicted. Collectively, results of this study support the idea that patterns of microbial diversity found in endolithic communities are governed by principles similar to those observed in macroecological systems.

Microbial diversity in the large intestine of pigs born and reared in different environments

Pigs born outdoors and reared on deep litter (straw) have been reported to experience less of a growth check after weaning and have a higher dressing percentage than counterparts born and reared under conventional (indoor) systems. The reason(s) for this difference is/are presently unknown, but differences in the gut environment might contribute to these observations. PCR-DGGE techniques were used in this study to examine microbial diversity and banding patterns in the large intestine of piglets that were reared under different rearing conditions. Six piglets per treatment were euthanised at weaning (21 days) and at 7 days and 21 days after weaning from two extreme treatments [indoor-born: conventionally-raised after weaning ('Indoor') or outdoor-born, deep-litter raised after weaning ('Outdoor')]. The Shannon diversity index was calculated, and multivariate analysis of banding patterns was performed. Indoor pigs had a more diverse bacterial population at weaning and 21 days after weaning than Outdoor pigs. However at the end of the first week after weaning, outdoor-born and deep-litter pigs had a more diverse microbiota. The Shannon diversity index continued to increase with time after weaning in Outdoor pigs, which did not occur in Indoor pigs. Multivariate analysis of banding patterns showed there was a trend (P = 0.109) for a difference in microbial structure depending on housing type. There was also a significant (P < 0.001) effect of sampling time after weaning and a significant interaction (P < 0.001) between housing and time after weaning.

Microbial communities in the larval midgut of laboratory and field populations of cotton bollworm (Helicoverpa armigera)

We compared the bacterial communities in the larval midgut of field and laboratory populations of a polyphagous pest, the cotton bollworm (Helicoverpa armigera), using denaturing gradient gel electrophoresis (DGGE) of amplified 16S rDNA sequences and 16S library sequence analysis. DGGE profiles and 16S rDNA library sequence analysis indicated similar patterns of midgut microbial community structure and diversity: specific bacterial types existed in both populations, and a more diverse microbial community was observed in caterpillars obtained from the field. The laboratory population harbored a rather simple gut microflora consisting mostly of phylotypes belonging to Enterococcus (84%). For the field population, phylotypes belonging to Enterococcus (28%) and Lactococcus (11%), as well as Flavobacterium (10%), Acinetobacter (19%), and Stenotrophomonas (10%) were dominant members. These results provided the first comprehensive description of the microbial diversity of the midgut of the important pest cotton bollworm and suggested that the environment and food supply might influence the diversity of the gut bacterial community.

Impact of carbon-rich dairy factory effluent on growth of perennial ryegrass ( Lolium perenne) and soil microorganisms

A pot experiment was conducted to investigate the impact of high carbon dairy factory effluent application on the growth of perennial ryegrass ( Lolium perenne L.), plant nutrient uptake, soil microbial biomass carbon and nitrogen, populations of soil-microorganisms, root colonising fungi and the microbial functional diversity. The effluent was added at rates of 0, 100,000, 200,000 and 300,000 l ha –1. These rates are equivalent to 0, × 1, × 2 and × 3 normal field application rates. The added effluent contained (g l –1), C; 19.42, total P; 0.65; S, 0.75, K; 1.33, Na; 4.55, Mg; 0.11, NH 4; 0.073, total N; 0.073 and had a pH of 4.33. Replicate pots (incubated in a controlled-environment room at 20 °C, with 16 h light/8 h dark) were harvested at 32, 61, and 130 days after setting up of the experiment. In the first sampling, shoot dry matter levels declined significantly ( P < 0.01) with increased effluent. By the third sampling the trend was reversed with treated pots having greater amounts of shoot dry matter. The initial depression of growth was possibly due to a combination of factors including excess levels of available carbon (C) for microbes leading to immobilisation of nutrients, particularly nitrogen (N) and sulphur (S). Shoot N and S concentrations were lower ( P < 0.001) and the phosphorus concentrations were higher in effluent-treated samples. Soil microbial biomass-C (480 and 770 μg g −1 of biomass C in untreated and treated soil, respectively) and microbial-N (81 and 123 μg g –1 of microbial-N in untreated and treated soil, respectively) were significantly ( P < 0.001) greater in effluent-treated pots at all times. Populations of total culturable bacteria were higher ( P < 0.01) in the treated pots in the first sample (log 10 populations g –1 were 7.3 in untreated pots compared to 8.0 averaged across three treatments) but there were no differences in the subsequent two samples. Effluent also increased yeast populations (log 10 numbers g –1 were 0.6 in untreated pots and 3.1 in treated pots averaged across treatments and times P < 0.01) at all three sampling times. The Shannon-Weiner Diversity Index of root fungi decreased with increasing effluent application ( P < 0.01) while the species richness decreased with effluent as well as with time ( P < 0.1). Potential root pathogens Fusarium oxysporum, total Fusarium spp. and Pythium spp. significantly increased ( P < 0.05) in treated samples but in the final sampling, Codinaea fertilis significantly ( P < 0.05) decreased with effluent treatment. The microbial functional diversity pattern and the average well colour development (AWCD) in soil were significantly changed by the effluent application but effects were not detectable after 130 days.

Coupling 16S‐ITS rDNA clone libraries and automated ribosomal intergenic spacer analysis to show marine microbial diversity: development and application to a time series

We outline an approach to simultaneously assess multilevel microbial diversity patterns utilizing 16S-ITS rDNA clone libraries coupled with automated ribosomal intergenic spacer analysis (ARISA). Sequence data from 512 clones allowed estimation of ARISA fragment lengths associated with bacteria in a coastal marine environment. We matched 92% of ARISA peaks (each comprising >1% total amplified product) with corresponding lengths from clone libraries. These peaks with putative identification accounted for an average of 83% of total amplified community DNA. At 16S rDNA similarities <98%, most taxa displayed differences in ARISA fragment lengths >10 bp, readily detectable and suggesting ARISA resolution is near the 'species' level. Prochlorococcus abundance profiles from ARISA were strongly correlated (r2=0.86) to Prochlorococcus cell counts, indicating ARISA data are roughly proportional to actual cell abundance within a defined taxon. Analysis of ARISA profiles for 42 months elucidated patterns of microbial presence and abundance providing insights into community shifts and ecological niches for specific organisms, including a coupling of ecological patterns for taxa within the Prochlorococcus, the Gamma Proteobacteria and Actinobacteria. Clade-specific ARISA protocols were developed for the SAR11 and marine cyanobacteria to resolve ambiguous identifications and to perform focused studies. 16S-ITS data allowed high-resolution identification of organisms by ITS sequence analysis, and examination of microdiversity.

Captured Diversity in a Culture Collection: Case Study of the Geographic and Habitat Distributions of Environmental Isolates Held at the American Type Culture Collection

The use of molecular techniques to assess prokaryotic diversity independent of the need for enrichment culture has profoundly changed how we view and study microbial diversity. As molecular data have accumulated over the past 15 years, these data are now resulting in a healthy debate about how much we really know about prokaryotic diversity in a wide range of environments (8, 21–23, 32, 44, 55). A number of recent articles have discussed these questions, and opinions range from the position that prokaryotic diversity is essentially infinite, with the existence of millions of potential species, (6, 10), to arguments that microbial diversity is reasonably finite (17, 26). In light of this debate concerning the results of theoretical considerations and molecule-based surveys, it is interesting to take stock of the holdings of the environmental prokaryotes available at the American Type Culture Collection (ATCC) to determine how these holdings reflect overall trends in microbial diversity in different habitats. The principle mission of bioresource centers, such as the ATCC, is to serve as living stock collections that acquire diverse biological materials for redistribution to researchers throughout the world. The ATCC has about two-thirds of the type strains of prokaryotic species in the world. In many cases, multiple strains of a given species are acquired. As these materials are accessioned, data relevant to their provenance and physiology are collected. These data are an underutilized asset of bioresource centers. For example, the data on the source locations of isolates can provide information about the relative sampling efforts for different geographic regions. Concordantly, data concerning the environmental habitats of isolates can provide important information about the diversity of genera that are associated with certain habitat types. No articles were found in a literature search that specifically analyzed culture collections as a metric for assessing our knowledge of larger patterns of microbial diversity. In this article we present an analysis of the holdings of environmental prokaryotes at the ATCC with regard to the geographical and environmental habitats from which they were isolated. The term environmental is broadly defined to include all organisms that are not pathogenic to humans or animals or that are not otherwise human associated (see below). Selected habitats for which holdings at the ATCC are the most abundant were compared to recently published findings of workers who used cultivation-independent methods to assess microbial diversity. This provided perspective on how well the “captured diversity” in a culture collection represents the diversity which may exist in nature.

Phytoplankton species richness scales consistently from laboratory microcosms to the world's oceans.

Species-area relationships have been observed for virtually all major groups of macroorganisms that have been studied to date but have not been explored for microscopic phytoplankton algae, which are the dominant producers in many freshwater and marine ecosystems. Our analyses of data from 142 different natural ponds, lakes, and oceans and 239 experimental ecosystems reveal a strong species-area relationship with an exponent that is invariant across ecosystems that span >15 orders of magnitude in spatial extent. A striking result is that the species-area relationship derived from small-scale experimental studies correctly scales up to natural aquatic ecosystems. These results significantly broaden our knowledge of the effects of island size on biodiversity and also confirm the relevance of experimentally derived data to the analysis and understanding of larger-scale ecological patterns. In addition, they confirm that patterns in microbial diversity are strongly consistent with those that have been repeatedly reported in the literature for macroorganisms.

Changes in microbial community metabolism and labile organic matter fractions as early indicators of the impact of management on soil biological quality

Changes to the metabolic profiles of soil microbial communities could have potential for use as early indicators of the impact of management or other perturbations on soil functioning and soil quality. We compared the relative susceptibility to management of microbial community metabolism with a number of soil organic matter (OM) and microbial parameters currently used as indicators of changes in soil biological quality. Following long-term cereal cropping, plots were subjected to a 16-month treatment period consisting of either a mixed cropping sequence of vetch, spring barley and clover or a continuous grass-clover ley which was periodically mown and mulched. The treatments had no effect on soil biomass N or respiration of microbial populations inoculated into Biolog Gram negative (GN) plates. After 16 months there were no management-induced changes to total OM, light-fraction OM C and N, labile organic N or water-soluble carbohydrates. However, patterns of substrate utilization by the soil microbial population following inoculation into Biolog GN plates were found to be highly sensitive to management practice. In the mixed cropping sequence, substrate utilization changed markedly following plough-in of the vetch crop, with a smaller change occurring after harvesting of the barley. In the ley treatment, substrate utilization was not affected until the onset of mowing, when the pattern changed to become similar to that in the mixed cropping sequence. Metabolic diversity of the Biolog-culturable microbial population was increased by the ley treatment, but was not affected by the cropping sequence. We conclude that patterns of microbial substrate utilization and metabolic diversity are more sensitive to the effects of management than are OM and biomass pools, and therefore have value as early indicators of the impacts of management on soil biological properties, and hence soil quality.

Development of a 5-step multi-chamber reactor as a simulation of the human intestinal microbial ecosystem.

A five-stage reactor was developed to simulate the gastro-intestinal microbial ecosystem of humans. The small intestine was simulated by a two-step "fill and draw" system, the large intestine by a three-step reactor. A representative supply medium was developed to support a microbial community resembling that of the human gastro-intestinal tract. The entire system was validated by monitoring fermentation fluxes and products, i.e. indicator bacterial groups, volatile fatty acids, enzymatic activities and headspace gases. The simulator was operated with varying concentrations and combinations of arabinogalactan, xylan, pectin, dextrins and starch. The resulting patterns of microbial diversity and activity were analysed and compared with data for in-vivo gastro-intestinal microbial communities as described in the literature and found to be representative.