Leaf-Associated Bacterial and Fungal Taxa Shifts in Response to Larvae of the Tree Hole Mosquito, <Emphasis Type="Italic">Ochlerotatus triseriatus</Emphasis>

Larvae of the eastern tree hole mosquito, Ochlerotatus triseriatus (Say), and related container-breeding species are known to feed upon substrate-associated microorganisms. Although the importance of these microbial resources to larval growth has been established, almost nothing is known about the taxonomic composition and dynamics of these critical microbial food sources. We examined bacterial and fungal community compositional changes on oak leaves tethered in natural tree hole habitats of O. triseriatus. We eliminated larvae experimentally in a subset of the tree holes and examined 16S rDNA gene sequences for bacteria and ergosterol concentrations and 18S rRNA gene sequences for fungi collected from leaf material subsamples. Leaf ergosterol content varied significantly with time, but not treatment. Principal component analysis (PCA) was used to compare microbial taxonomic patterns found in leaves incubated with or without larvae present, and we found that larval presence affected both bacterial and fungal groups, either from loosely attached or strongly adherent categories. Bacterial communities generally grouped more tightly when larvae were present, and class level taxa proportions changed when larvae were present, suggesting selection by larval feeding or activities for particular taxa such as members of the Bacteroidetes, Alphaproteobacteria, and Betaproteobacteria classes. Fungal taxa composite scores also separated along PC axes related to the presence of larvae and indicated larval feeding effects on several higher taxonomic groups, including Saccharomycetes, Dothideomycetes, and Chytridiomycota. These results support the hypothesis that larval mosquito feeding and activities altered microbial communities associated with substrate surfaces, potentially leading to decreased food value of the resource and affecting decomposition of particulate matter in the system.     

Aquatic microfauna alter larval food resources and affect development and biomass of West Nile and Saint Louis encephalitis vector Culex nigripalpus (Diptera: Culicidae)

Ciliate protists and rotifers are ubiquitous in aquatic habitats and can comprise a significant portion of the microbial food resources available to larval mosquitoes, often showing substantial declines in abundance in the presence of mosquito larvae. This top‐down regulation of protists is reported to be strong for mosquitoes inhabiting small aquatic containers such as pitcher plants or tree holes, but the nature of these interactions with larval mosquitoes developing in other aquatic habitats is poorly understood. We examined the effects of these two microbial groups on lower trophic level microbial food resources, such as bacteria, small flagellates, and organic particles, in the water column, and on Culex larval development and adult production. In three independent laboratory experiments using two microeukaryote species (one ciliate protist and one rotifer) acquired from field larval mosquito habitats and cultured in the laboratory, we determined the effects of Culex nigripalpus larval grazing on water column microbial dynamics, while simultaneously monitoring larval growth and development. The results revealed previously unknown interactions that were different from the top‐down regulation of microbial groups by mosquito larvae in other systems. Both ciliates and rotifers, singly or in combination, altered other microbial populations and inhibited mosquito growth. It is likely that these microeukaryotes, instead of serving as food resources, competed with early instar mosquito larvae for microbes such as small flagellates and bacteria in a density‐dependent manner. These findings help our understanding of the basic larval biology of Culex mosquitoes, variation in mosquito production among various larval habitats, and may have implications for existing vector control strategies and for developing novel microbial‐based control methods.     

Phagotrophic protists are a key component of microbial communities processing leaf litter under contrasting oxic conditions

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Leaf litter species identity alters the structure of pond communities

The input of leaf litter resources is a major driver of ecosystem processes in terrestrial and freshwater habitats. Although variation exists in the quantity and composition of litter inputs due to natural and anthropogenic causes, few studies have examined how such variation influences the structure and composition of aquatic food webs. Using outdoor mesocosms, we examined the bottom–up effects of 10 chemically distinct tree litter species on microbial, algal, invertebrate and vertebrate fauna found in temperate ponds. We hypothesized that individual litter species, which differ in their traits, would differentially and predictably affect abiotic and biotic elements of pond communities. We further hypothesized that the presence of leaf litter, regardless of species, would elevate resource supply and increase the biomass of community members. Finally, we hypothesized that a mixture of litter species would have non‐additive effects on community responses. We followed the system for > 4 months and measured > 30 abiotic and biotic responses related to primary and secondary production. The different species of leaf litter had major effects on abiotic and biotic responses, including phytoplankton, periphyton, zooplankton, snails, amphipods and tadpoles. Most biological responses were negatively associated with soluble carbon content of litter, or litter decay rate. Other litter traits, including phenolic concentrations and litter C:N were of secondary importance but did exhibit both positive and negative associations with several responses. The absence of litter had pervasive effects on abiotic attributes, but did not promote substantial changes in organism biomass. Most responses to the litter mixture were additive. Our results suggest that changes in temperate forest composition can strongly affect pond communities.     

Bacterial microbiota assemblage in Aedes albopictus mosquitoes and its impacts on larval development

Interactions between bacterial microbiota and mosquitoes play an important role in mosquitoes’ capacity to transmit pathogens. However, microbiota assemblages within mosquitoes and the impact of microbiota in environments on mosquito development and survival remain unclear. This study examined microbiota assemblages and the effects of aquatic environment microbiota on the larval development of the Aedes albopictus mosquito, an important dengue virus vector. Life table studies have found that reducing bacterial load in natural aquatic habitats through water filtering and treatment with antibiotics significantly reduced the larva‐to‐adult emergence rate. This finding was consistent in two types of larval habitats examined—discarded tires and flowerpots, suggesting that bacteria play a crucial role in larval development. Pyrosequencing of the bacterial 16S rRNA gene was used to determine the diversity of bacterial communities in larval habitats and the resulting numbers of mosquitoes under both laboratory and field conditions. The microbiota profiling identified common shared bacteria among samples from different years; further studies are needed to determine whether these bacteria represent a core microbiota. The highest microbiota diversity was found in aquatic habitats, followed by mosquito larvae, and the lowest in adult mosquitoes. Mosquito larvae ingested their bacterial microbiota and nutrients from aquatic habitats of high microbiota diversity. Taken together, the results support the observation that Ae. albopictus larvae are able to utilize diverse bacteria from aquatic habitats and that live bacteria from aquatic habitats play an important role in larval mosquito development and survival. These findings provide new insights into bacteria's role in mosquito larval ecology.     

Effects of burial on leaf litter quality,microbial conditioning and palatability to three shredder taxa

Summary 1. Heterotrophic microorganisms are crucial for mineralising leaf litter and rendering it more palatable to leaf‐shredding invertebrates. A substantial part of leaf litter entering running waters may be buried in the streambed and thus be exposed to the constraining conditions prevailing in the hyporheic zone. The fate of this buried organic matter and particularly the role of microbial conditioning in this habitat remain largely unexplored. 2. The aim of this study was to determine how the location of leaf litter within the streambed (i.e. at the surface or buried), as well as the leaf litter burial history, may affect the leaf‐associated aquatic hyphomycete communities and therefore leaf consumption by invertebrate detritivores. We tested the hypotheses that (i) burial of leaf litter would result in lower decomposition rates associated with changes in microbial assemblages compared with leaf litter at the surface and (ii) altered microbial conditioning of buried leaf litter would lead to decreased quality and palatability to their consumers, translating into lower growth rates of detritivores. 3. These hypotheses were tested experimentally in a second‐order stream where leaf‐associated microbial communities, as well as leaf litter decomposition rates, elemental composition and toughness, were compared across controlled treatments differing by their location within the streambed. We examined the effects of the diverse conditioning treatments on decaying leaf palatability to consumers through feeding trials on three shredder taxa including a freshwater amphipod, of which we also determined the growth rate. 4. Microbial leaf litter decomposition, fungal biomass and sporulation rates were reduced when leaf litter was buried in the hyporheic zone. While the total species richness of fungal assemblages was similar among treatments, the composition of fungal assemblages was affected by leaf litter burial in sediment. 5. Leaf litter burial markedly affected the food quality (especially P content) of leaf material, probably due to the changes in microbial conditioning. Leaf litter palatability to shredders was highest for leaves exposed at the sediment surface and tended to be negatively related to leaf litter toughness and C/P ratio. In addition, burial of leaf litter led to lower amphipod growth rates, which were positively correlated with leaf litter P content. 6. These results emphasise the importance of leaf colonisation by aquatic fungi in the hyporheic zone of headwater streams, where fungal conditioning of leaf litter appears particularly critical for nutrient and energy transfer to higher trophic levels.     

Seasonal availability of resources and habitat degradation for the western tree-hole mosquito, <Emphasis Type="Italic">Aedes sierrensis</Emphasis>

The nutrient base of aquatic tree-hole communities is derived from leaf litter, benthic detritus, and water flowing down the tree trunk (stemflow water). Previous studies in eastern North America with the mosquito, Aedes triseriatus, have identified leaf litter as a major and stemflow water as a minor source of mosquito nutrition, but did not consider the role of the benthic detritus or how the aggregate or relative contribution of these sources of mosquito nutrition changed during the year. We use the leaf litter, benthic detritus, and stemflow water from tree holes in western Oregon (USA) to determine how these substrates affect mass at metamorphosis, biomass yield, and fitness (cohort replacement rate; R ₀) of the mosquito, Aedes sierrensis, through both natural and simulated winters, the normal growing season for larvae in tree holes. We found that fresh leaf litter constitutes the major determinant of mosquito fitness by a factor of >15:1 over any other substrate taken directly from tree holes in nature. The other substrates, including the benthic detritus, individually make only a meager contribution to mosquito fitness but, when added to the leaf litter, can sustain yield and improve fitness at high, limiting larval densities. Nutritional quality of tree-hole substrates declines by >90% from early (fall) to late (spring) in the larval growing season. At both times of year, the coarse or fine detritus provide minor resources, and stemflow water provides no detectable contribution to mosquito nutrition. The resources in the litter are not transported during the year to the benthic detritus; rather, these resources are either exploited by mosquitoes when they first become available, or they deteriorate and become progressively more unavailable to them. Growth and development of A. sierrensis feeding on dried and reconstituted tree-hole contents during a 6-month simulated winter in the laboratory showed: (1) the same relative contributions of leaf litter, benthic detritus, and stemflow water to mosquito nutrition, (2) that the winter deterioration of substrate quality is a direct consequence of microbial decomposition, and (3) that pre-emptive competition from pre-existing A. sierrensis greatly increases substrate deterioration. We conclude that the progressive winter deterioration of larval resources in combination with the dry summers of western North America are the most likely environmental factors that limit species diversity in tree holes and that have selected for early recruitment (autumnal hatching) of A. sierrensis and for its univoltine life cycle from Mexico to Canada. Received: 28 December 1999 / Accepted: 10 April 2000     

Nestedness patterns of container-dwelling mosquitoes: effects of larval habitat within variable terrestrial matrices

Distributions of mosquito larvae likely are a consequence of multiple factors, although two commonly studied factors (quality of the larval environment and the terrestrial matrix in which these habitats reside) have rarely and simultaneously been varied in the field to understand distributions of larvae. We monitored aquatic containers of two leaf detritus levels within a forest, prairie, and industrial habitat across five months to understand the temporal and spatial colonization of aquatic invertebrates in Northcentral Illinois, USA. Data were collected monthly on mosquito populations and the composition of other invertebrates colonizing containers. Overall, six species of mosquitoes colonized containers, with Culex restuans and Aedes triseriatus having the highest relative abundances. There were strong seasonal abundance patterns for these two mosquito species, with the dominant species changing over time in the forest habitat. The responses of other mosquito taxa were more variable, with abundances reflective of either the terrestrial matrix or larval habitat quality. High detritus containers supported the highest abundances of most species encountered, regardless of habitat. Non-mosquito taxa were less common numerically, but analyses suggested that some taxa, such as syrphid larvae, often co-occurred with mosquitoes. Nested subset analysis indicated communities were strongly nested, and that both habitat type and detritus level were important in explaining nested patterns of aquatic invertebrates. Our data show that both the larval habitat and the surrounding terrestrial matrix shape patterns of container mosquitoes, and that other container invertebrates vary in similar ways as mosquitoes. Handling editor: K. Martens     

EFFECTS OF ELEVATED ATMOSPHERIC CO(2) ON WATER CHEMISTRY AND MOSQUITO (DIPTERA: CULICIDAE) GROWTH UNDER COMPETITIVE CONDITIONS IN CONTAINER HABITATS

We investigated the direct and indirect effects of elevated atmospheric CO(2) on freshwater container habitats and their larval mosquito occupants. We predicted that a doubling of atmospheric CO(2) would (1) alter the chemical properties of water in this system, (2) slow degradation of leaf litter, and (3) decrease larval growth of Aedes albopictus (Skuse) mosquitoes raised on that litter under competitive conditions. Effects of elevated CO(2) on water quality parameters were not detected, but the presence of leaf litter significantly reduced pH and dissolved oxygen relative to water-filled containers without litter. Degradation rates of oak leaf litter from plants grown under elevated CO(2) atmospheres did not differ from breakdown rates of litter from ambient CO(2) conditions. Litter from plants grown in an elevated CO(2) atmospheres did not influence mosquito population growth, but mosquito production decreased significantly with increasing larval density. Differences among mosquito density treatments influenced survivorship most strongly among male Ae. albopictus and time to emergence most strongly among females, suggesting fundamental sex-determined differences in response to competition. Results of this and other studies indicate that direct and indirect effects of doubled atmospheric CO(2) are minimal in artificial containers with freshwater.     

Does mixing litter of different qualities alter stream microbial diversity and functioning on individual litter species?

We examined effects of leaf litter quality and species mixing on microbial community diversity and litter processing in a forested headwater stream. Single- and mixed-species litter from dominant tree species ( Liriodendron tulipifera , Acer rubrum , Quercus prinus , Rhododendron maximum ) were incubated in a southern Appalachian headwater stream. Litter carbon-to-nitrogen ratios (C:N), mass loss, microbial respiration, and microbial community diversity were analyzed on individual litter species after incubation. Initial C:N varied widely among individual litter species, and these differences persisted throughout the 50-day incubation period. Litter C:N of the recalcitrant species R. maximum remained higher than that of all other litter species, and C:N of R. maximum and L. tulipifera increased when both species were present together in a mixture. Although mass loss of individual species was generally unaffected by mixing, microbial respiration was greater on A. rubrum and Q. prinus litter incubated with R. maximum compared to either species alone. Enhanced resource heterogeneity, which was experimentally achieved by litter mixing low- and higher-quality litter species, resulted in apparent shifts in microbial community diversity on individual litter species. Responses of bacterial and fungal community diversity to litter mixing varied among individual litter species. Our results suggest that changes in tree species composition in riparian forests and subsequent changes in litter resource heterogeneity could alter stream microbial community diversity and function. As bacteria and fungi are important decomposers of plant litter in aquatic ecosystems, resource-dependent changes in microbial communities could alter detrital processing dynamics in streams.     

Increased Larval Mosquito Densities from Modified Landuses in the Kapiti Region,New Zealand: Vegetation,Water Quality,and Predators as Associated Environmental Factors

Landuse changes, including deforestation, agriculture, and urbanization, have coincided with an increase in vector-borne diseases worldwide. Landuse changes may alter mosquito populations by modifying the characteristics of aquatic larval habitats, but we still poorly understand the physical, chemical, and biological factors involved. We examined a total of 81 mosquito larval habitats for immature mosquitoes and 17 environmental variables in native forest, pastureland, and urbanland, at three locations in the Kapiti region, New Zealand. Significantly higher immature mosquito densities, predominantly of the endemic species Cx. pervigilans, were collected from urbanland and pastureland compared to native forest. Urbanland and pastureland habitats were mostly artificial containers compared to ground pools in native forest. Generalized linear modeling (GLM) revealed nine environmental variables that were significantly different between landuses. Of these variables, mosquito density was significantly (positively) correlated with bacteria and dissolved organic carbon. When location and date were controlled for in GLM, mosquito density was (negatively) related to the presence of vegetation and combined predators. The findings of this study support those from prior surveys in warmer climates suggesting greater mosquito-borne disease risk in anthropogenically-modified environments because of ecosystem disruption. Unlike most previous field-based work, this study suggests that in addition to habitat type, the presence of vegetation, water quality, and predators are also associated with mosquito density and may be involved in causal mechanisms. Urban containers and stock drinking troughs had high mosquito densities, suggesting that an initial step in directing control operations should be to focus on these habitats.     

Living in the litter: the influence of tree leaf litter on wetland communities

Empirical research in streams has demonstrated that terrestrial subsidies of tree leaf litter influence multiple community factors including composition, diversity and growth of individuals. However, little research has examined the importance of tree litter species on wetlands, which are ubiquitous across the landscape and serve as important habitats for a unique and diverse community of organisms. Using outdoor mesocosms, we assessed the impact of 12 litter monocultures and three litter mixtures (from both broadleaf and conifer trees) on pond communities containing gray tree frog tadpoles Hyla versicolor, periphyton, phytoplankton and zooplankton. We found that leaf litter species had substantial and differential impacts on all trophic groups in the community including effects on algal abundance, zooplankton density and amphibian growth. In many instances, patterns of responses were specific to individual litter species yet some responses, including both pH values and periphyton biomass, were generalizable to broad taxonomic groups. In addition, while most responses of litter mixtures were additive, we found evidence for antagonistic effects of litter mixing among responses of periphyton and amphibian body mass. Our results highlight the potential impact of human and naturally driven changes in forest composition on wetland communities through associated changes in leaf litter.     

Evaluation of macrofaunal effects on leaf litter breakdown rates in aquatic and terrestrial habitats

Abstract Decomposition of the organic matter is a key process in the functioning of aquatic and terrestrial ecosystems, although different factors influence processing rates between and within these habitats. Most patterns were described for temperate regions, with fewer studies in tropical, warmer sites. In this study, we carried out a factorial experiment to compare processing rates of mixed species of leaf litter between terrestrial and aquatic habitats at a tropical site, using ?ne and coarse mesh cages to allow or prevent colonization by macroinvertebrates. The experiment was followed for 10 weeks, and loss of leaf litter mass through time was evaluated using exponential models. We found no interaction between habitat and mesh size and leaf litter breakdown rates did not differ between ?ne and coarse mesh cages, suggesting that macroinvertebrates do not influence leaf litter decomposition in either habitat at our studied site. Leaf breakdown rates were faster in aquatic than in terrestrial habitats and the magnitude of these differences were comparable to studies in temperate regions, suggesting that equivalent factors can influence between‐habitat differences detected in our study.     

Leaf species identity and combination affect performance and oviposition choice of two container mosquito species

Abstract 1. Resource diversity can be an important determinant of individual and population performance in insects. Fallen parts of plants form the nutritive base for many aquatic systems, including mosquito habitats, but the effect of plant diversity on mosquito production is poorly understood. 2. To determine the effects of diverse plant inputs on larval mosquitoes, experiments were conducted that examined how leaves of Vitis aestivalis, Quercus virginiana, Psychotria nervosa, and Nephrolepis exaltata affected the container species Aedes triseriatus and Aedes albopictus. 3. The hypothesis that leaf species have different effects on larval survival, growth, population performance, and oviposition choice of the two mosquito species was tested. The hypothesis that larval performance of A. albopictus responds additively to combinations of the four plant species was also tested. 4. Larval survival and growth differed among the four leaf species, and oviposition preference differed among the two leaf species examined. Measurements of population performance demonstrated significant variation between leaf treatments. Larval outcomes for A. albopictus were significantly affected by leaf combination, and the hypothesis of additivity could be rejected. 5. These results indicate that individual leaf species are important in determining the performance of container dwelling mosquitoes, which grow larger and survive better on mixed‐species resource than expected, based on an additive model of resource utilisation.     

Leaf Litter Mixtures Alter Microbial Community Development: Mechanisms for Non-Additive Effects in Litter Decomposition

To what extent microbial community composition can explain variability in ecosystem processes remains an open question in ecology. Microbial decomposer communities can change during litter decomposition due to biotic interactions and shifting substrate availability. Though relative abundance of decomposers may change due to mixing leaf litter, linking these shifts to the non-additive patterns often recorded in mixed species litter decomposition rates has been elusive, and links community composition to ecosystem function. We extracted phospholipid fatty acids (PLFAs) from single species and mixed species leaf litterbags after 10 and 27 months of decomposition in a mixed conifer forest. Total PLFA concentrations were 70% higher on litter mixtures than single litter types after 10 months, but were only 20% higher after 27 months. Similarly, fungal-to-bacterial ratios differed between mixed and single litter types after 10 months of decomposition, but equalized over time. Microbial community composition, as indicated by principal components analyses, differed due to both litter mixing and stage of litter decomposition. PLFA biomarkers a15∶0 and cy17∶0, which indicate gram-positive and gram-negative bacteria respectively, in particular drove these shifts. Total PLFA correlated significantly with single litter mass loss early in decomposition but not at later stages. We conclude that litter mixing alters microbial community development, which can contribute to synergisms in litter decomposition. These findings advance our understanding of how changing forest biodiversity can alter microbial communities and the ecosystem processes they mediate.     

Fungal communities on decaying leaves in streams: a comparison of two leaf species

Decomposition of leaf litter is a microbial mediated process that helps to transfer energy and nutrients from leaves to higher trophic levels in woodland streams. Generally, aquatic hyphomycetes are viewed as the major fungal group responsible for leaf litter decomposition. In this study, traditional microscopic examination (based on identification of released conidia) and phylogenetic analysis of 18S rRNA genes from cultivated fungi were used to compare fungal community composition on decomposing leaves of two species (sugar maple and white oak) from a NE Ohio stream. No significant differences were found in sporulation rates between maple and oak leaves and both had similar species diversity. From the 18S rRNA gene sequence data, identification was achieved for 12 isolates and taxonomic affiliation of 12 of the remaining 14 isolates could be obtained. A neighbor-joining tree (with bootstrap values) was constructed to examine the taxonomic distribution of the isolates relative to sequences of known operational taxonomic units (OTUs). Surprisingly, only 2 of the isolates obtained were aquatic hyphomycetes based on phylogenetic analysis. Overall, there were no differences between the two leaf types and a higher diversity was observed via culturing and subsequent 18S rRNA gene sequencing than by conidia staining. These differences resulted from the fact that traditional microscopy provides estimates of aquatic hyphomycete diversity while the other approach revealed the presence of both aquatic hyphomycete and non-aquatic hyphomycete taxa. The presence of this broad array of taxa suggests that the role of aquatic hyphomycetes relative to other fungi be re-evaluated. Even though the functional role of these non-aquatic hyphomycetes taxa is unknown, their presence and diversity demonstrates the need to delve further into fungal community structure on decomposing leaves.     

Elevated temperature may reduce functional but not taxonomic diversity of fungal assemblages on decomposing leaf litter in streams

Mounting evidence points to a linkage between biodiversity and ecosystem functioning (B-EF). Global drivers, such as warming and nutrient enrichment, can alter species richness and composition of aquatic fungal assemblages associated with leaf-litter decomposition, a key ecosystem process in headwater streams. However, effects of biodiversity changes on ecosystem functions might be countered by the presumed high functional redundancy of fungal species. Here, we examined how environmental variables and leaf-litter traits (based on leaf chemistry) affect taxonomic and functional α- and β-diversity of fungal decomposers. We analysed taxonomic diversity (DNA-fingerprinting profiles) and functional diversity (community-level physiological profiles) of fungal communities in four leaf-litter species from four subregions differing in stream-water characteristics and riparian vegetation. We hypothesized that increasing stream-water temperature and nutrients would alter taxonomic diversity more than functional diversity due to the functional redundancy among aquatic fungi. Contrary to our expectations, fungal taxonomic diversity varied little with stream-water characteristics across subregions, and instead taxon replacement occurred. Overall taxonomic β-diversity was fourfold higher than functional diversity, suggesting a high degree of functional redundancy among aquatic fungi. Elevated temperature appeared to boost assemblage uniqueness by increasing β-diversity while the increase in nutrient concentrations appeared to homogenize fungal assemblages. Functional richness showed a negative relationship with temperature. Nonetheless, a positive relationship between leaf-litter decomposition and functional richness suggests higher carbon use efficiency of fungal communities in cold waters.     

Placing the Effects of Leaf Litter Diversity on Saprotrophic Microorganisms in the Context of Leaf Type and Habitat

Because of conflicting results in previous studies, it is unclear whether litter diversity has a predictable impact on microbial communities or ecosystem processes. We examined whether effects of litter diversity depend on factors that could confound comparisons among previous studies, including leaf type, habitat type, identity of other leaves in the mixture, and spatial covariance at two scales within habitats. We also examined how litter diversity affects the saprotrophic microbial community using terminal restriction fragment length polymorphism to profile bacterial and fungal community composition, direct microscopy to quantify bacterial biomass, and ergosterol extraction to quantify fungal biomass. We found that leaf mixture diversity was rarely significant as a main effect (only for fungal biomass), but was often significant as an interaction with leaf type (for ash-free dry mass recovered, carbon-to-nitrogen ratio, fungal biomass, and bacterial community composition). Leaf type and habitat were significant as main effects for all response variables. The majority of variance in leaf ash-free dry mass and C/N ratio was explained after accounting for treatment effects and spatial covariation at the meter (block) and centimeter (litterbag) scales. However, a substantial amount of variability in microbial communities was left unexplained and must be driven by factors at other spatial scales or more complex spatiotemporal dynamics. We conclude that litter diversity effects are primarily dependent on leaf type, rather than habitat type or identity of surrounding leaves, which can guide the search for mechanisms underlying effects of litter diversity on ecosystem processes.     

Physiological Profiling of Soil Microbial Communities in a Florida Scrub-Oak Ecosystem: Spatial Distribution and Nutrient Limitations

Rapid physiological profiling of heterotrophic microbial communities enables intensive analysis of the factors affecting activity in aerobic habitats, such as soil. Previous methods for performing such profiling were severely limited due to enrichment bias and inflexibility in incubation conditions. We tested a new physiological profiling approach based on a microtiter plate oxygen sensor system (Becton Dickinson Oxygen Biosensor System (BDOBS)), which allows for testing of lower substrate addition (i.e., lower enrichment potential) and manipulation of physiochemical assay conditions, such as pH and nutrients. Soil microbial communities associated with a scrub-oak forest ecosystem on Merritt Island Wildlife Refuge in central Florida, USA, were studied in order to evaluate microbial activity in a nutrient poor soil and to provide baseline data on the site for subsequent evaluation of the effects of elevated CO₂ on ecosystem function. The spatial variation in physiological activity amongst different habitats (litter, bulk soil, and rhizosphere) was examined as a function of adaptation to local resources (i.e., water soluble extracts of roots and leaf litter) and the degree of N and P limitation. All the communities were primarily N-limited, with a secondary P limitation, which was greater in the rhizosphere and bulk soil. The litter community showed greater overall oxygen consumption when exposed to litter extracts relative to the rhizosphere or soil, suggesting acclimation toward greater use of the mixed substrates in the extract. Root extracts were readily used by communities from all the habitats with no habitat specific acclimation observed. A priming effect was detected in all habitats; addition of glucose caused a significant increase in the use of soil organic carbon. Response to added glucose was only observed with N and P addition, suggesting that C may be lost to the groundwater from these porous soils because nutrient limitation prevents C immobilization.     

Plant Communities, Soil Microorganisms, and Soil Carbon Cycling: Does Altering the World Belowground Matter to Ecosystem Functioning?

Soil microorganisms mediate many critical ecosystem processes. Little is known, however, about the factors that determine soil microbial community composition, and whether microbial community composition influences process rates. Here, we investigated whether aboveground plant diversity affects soil microbial community composition, and whether differences in microbial communities in turn affect ecosystem process rates. Using an experimental system at La Selva Biological Station, Costa Rica, we found that plant diversity (plots contained 1, 3, 5, or > 25 plant species) had a significant effect on microbial community composition (as determined by phospholipid fatty acid analysis). The different microbial communities had significantly different respiration responses to 24 labile carbon compounds. We then tested whether these differences in microbial composition and catabolic capabilities were indicative of the ability of distinct microbial communities to decompose different types of litter in a fully factorial laboratory litter transplant experiment. Both microbial biomass and microbial community composition appeared to play a role in litter decomposition rates. Our work suggests, however, that the more important mechanism through which changes in plant diversity affect soil microbial communities and their carbon cycling activities may be through alterations in their abundance rather than their community composition.