Daniel F. R. Cleary

Denaturing gradient gel electrophoresis and barcoded pyrosequencing reveal unprecedented archaeal diversity in mangrove sediment and rhizosphere samples.

ABSTRACT Mangroves are complex ecosystems that regulate nutrient and sediment fluxes to the open sea. The importance of bacteria and fungi in regulating nutrient cycles has led to an interest in their diversity and composition in mangroves. However, very few studies have assessed Archaea in mangroves, and virtually nothing is known about whether mangrove rhizospheres affect archaeal diversity and composition. Here, we studied the diversity and composition of Archaea in mangrove bulk sediment and the rhizospheres of two mangrove trees, Rhizophora mangle and Laguncularia racemosa, using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing of archaeal 16S rRNA genes with a nested-amplification approach. DGGE profiles revealed significant structural differences between bulk sediment and rhizosphere samples, suggesting that roots of both mangrove species influence the sediment archaeal community. Nearly all of the detected sequences obtained with pyrosequencing were identified as Archaea, but most were unclassified at the level of phylum or below. Archaeal richness was, furthermore, the highest in the L. racemosa rhizosphere, intermediate in bulk sediment, and the lowest in the R. mangle rhizosphere. This study shows that rhizosphere microhabitats of R. mangle and L. racemosa, common plants in subtropical mangroves located in Rio de Janeiro, Brazil, hosted distinct archaeal assemblages.

BIRD SPECIES AND TRAITS ASSOCIATED WITH LOGGED AND UNLOGGED FOREST IN BORNEO

The ecological consequences of logging have been and remain a focus of considerable debate. In this study, we assessed bird species composition within a logging concession in Central Kalimantan, Indonesian Borneo. Within the study area (approximately 196 km2) a total of 9747 individuals of 177 bird species were recorded. Our goal was to identify associations between species traits and environmental variables. This can help us to understand the causes of disturbance and predict whether species with given traits will persist under changing environmental conditions. Logging, slope position, and a number of habitat structure variables including canopy cover and liana abundance were significantly related to variation in bird composition. In addition to environmental variables, spatial variables also explained a significant amount of variation. However, environmental variables, particularly in relation to logging, were of greater importance in structuring variation in composition. Environmental change following logging appeared to have a pronounced effect on the feeding guild and size class structure but there was little evidence of an effect on restricted range or threatened species although certain threatened species were adversely affected. For example, species such as the terrestrial insectivore Argusianus argus and the hornbill Buceros rhinoceros, both of which are threatened, were rare or absent in recently logged forest. In contrast, undergrowth insectivores such as Orthotomus atrogularis and Trichastoma rostratum were abundant in recently logged forest and rare in unlogged forest. Logging appeared to have the strongest negative effect on hornbills, terrestrial insectivores, and canopy bark-gleaning insectivores while moderately affecting canopy foliage-gleaning insectivores and frugivores, raptors, and large species in general. In contrast, undergrowth insectivores responded positively to logging while most understory guilds showed little pronounced effect. Despite the high species richness of logged forest, logging may still have a negative impact on extant diversity by adversely affecting key ecological guilds. The sensitivity of hornbills in particular to logging disturbance may be expected to alter rainforest dynamics by seriously reducing the effective seed dispersal of associated tree species. However, logged forest represents an increasingly important habitat for most bird species and needs to be protected from further degradation. Biodiversity management within logging concessions should focus on maintaining large areas of unlogged forest and mitigating the adverse effects of logging on sensitive groups of species.

Taking root: enduring effect of rhizosphere bacterial colonization in mangroves.

Background Mangrove forests are of global ecological and economic importance, but are also one of the worlds most threatened ecosystems. Here we present a case study examining the influence of the rhizosphere on the structural composition and diversity of mangrove bacterial communities and the implications for mangrove reforestation approaches using nursery-raised plants. Methodology/Principal Findings A barcoded pyrosequencing approach was used to assess bacterial diversity in the rhizosphere of plants in a nursery setting, nursery-raised transplants and native (non-transplanted) plants in the same mangrove habitat. In addition to this, we also assessed bacterial composition in the bulk sediment in order to ascertain if the roots of mangrove plants affect sediment bacterial composition. We found that mangrove roots appear to influence bacterial abundance and composition in the rhizosphere. Due to the sheer abundance of roots in mangrove habitat, such an effect can have an important impact on the maintenance of bacterial guilds involved in nutrient cycling and other key ecosystem functions. Surprisingly, we also noted a marked impact of initial nursery conditions on the rhizosphere bacterial composition of replanted mangrove trees. This result is intriguing because mangroves are periodically inundated with seawater and represent a highly dynamic environment compared to the more controlled nursery environment. Conclusions/Significance In as far as microbial diversity and composition influences plant growth and health, this study indicates that nursery conditions and early microbial colonization patterns of the replants are key factors that should be considered during reforestation projects. In addition to this, our results provide information on the role of the mangrove rhizosphere as a habitat for bacteria from estuarine sediments.

Assessment of variation in bacterial composition among microhabitats in a mangrove environment using DGGE fingerprints and barcoded pyrosequencing.

Here, we use DGGE fingerprinting and barcoded pyrosequencing data, at six cut-off levels (85–100%), of all bacteria, Alphaproteobacteria and Betaproteobacteria to assess composition in the rhizosphere of nursery plants and nursery-raised transplants, native plants and bulk sediment in a mangrove habitat. When comparing compositional data based on DGGE fingerprinting and barcoded pyrosequencing at different cut-off levels, all revealed highly significant differences in composition among microhabitats. Procrustes superimposition revealed that ordination results using cut-off levels from 85–100% and DGGE fingerprint data were highly congruent with the standard 97% cut-off level. The various approaches revealed a primary gradient in composition from nursery to mangrove samples. The affinity between the nursery and transplants was greatest when using Betaproteobacteria followed by Alphaproteobacteria data. There was a distinct secondary gradient in composition from transplants to bulk sediment with native plants intermediate, which was most prevalent using all bacteria at intermediate cut-off levels (92–97%). Our results show that PCR-DGGE provides a robust and cost effective exploratory approach and is effective in distinguishing among a priori defined groups.

Coral diversity across a disturbance gradient in the Pulau Seribu reef complex off Jakarta, Indonesia

Very few coral reefs are located close enough to metropolitan cities to study the influence of large urban populations on reef communities. Here, we compare the impact of a large-scale disturbance gradient with local-scale disturbance on coral richness, cover, and composition in the Jakarta Bay and Pulau Seribu reef complex off Jakarta, Indonesia. We found no effect of local land-use type of coral reef islands on richness, composition or cover, nor did taxon richness differ among zones at the large-scale. There was, however, a pronounced difference in composition and coral cover among zones. Cover was very low and composition differed markedly in the near-shore zone 1 (Jakarta Bay) where human-induced disturbance is most intense. Cover was highest in the outlying reefs of zone 3. The highly perturbed zone 1 reefs were, furthermore, distinguished by the virtual absence of otherwise abundant coral taxa such as Acropora hyacinthus and Porites rus and the prevalence of taxa such as Oulastrea crispata and Favia maxima. Almost 60% of the spatial variation in composition was related to variation in shelf depth and island size. The importance of shelf depth was related to the prevalence of a strong environmental gradient in reef depth, pollution, and mechanical reef disturbance and salinity from Jakarta Bay to the outlying reefs. Although there was a significant univariate relationship between spatial variation in composition and distance, this did not enter into the multivariate model, except when presence–absence data was used, indicating that environmental processes are the primary structuring forces in determining local coral assemblage composition across the Pulau Seribu complex.

Habitat- and host-related variation in sponge bacterial symbiont communities in Indonesian waters

Marine lakes are unique ecosystems that contain isolated populations of marine organisms. Isolated from the surrounding marine habitat, many lakes house numerous endemic species. In this study, microbial communities of sponges inhabiting these lakes were investigated for the first time using barcoded pyrosequencing of 16S rRNA gene amplicons. Our main goals were to compare the bacterial richness and composition of two sponge species (Suberites diversicolor and Cinachyrella australiensis) inhabiting both marine lakes and adjacent open coastal systems. Host species and habitat explained almost 59% of the variation in bacterial composition. There was a significant difference in composition between both host species. Within S. diversicolor, there was little discernible difference between bacterial communities inside and outside lakes. The bacterial community of this species was, furthermore, dominated (63% of all sequences) by three very closely related alphaproteobacterial taxa identified as belonging to the recently described order Kiloniellales. Cinachyrella australiensis, in contrast, hosted markedly different bacterial communities inside and outside lakes with very few shared abundant taxa. Cinachyrella australiensis in open habitat only shared 9.4% of OTUs with C. australiensis in lake habitat. Bacteria were thus both highly species specific and, in the case of C. australiensis, habitat specific.

RANGE-RESTRICTED, SPECIALIST BORNEAN BUTTERFLIES ARE LESS LIKELY TO RECOVER FROM ENSO-INDUCED DISTURBANCE

The forest fires induced by the El Niño Southern Oscillation (ENSO) in 1997-1998 resulted in the temporary extirpation of more than 100 lowland butterfly species at a forest site in Borneo. Species with more restricted ranges were less likely to recover over the following four years. Matched-pair analyses revealed that species with lower initial abundances, restricted geographic ranges, and more specialized larvae were less likely to return. Specialization differed predictably between the (more generalist) wide-range and (more specialized) restricted-range species in our data set, and both geographic range and level of specialization were important in multivariate models. These are the first observations directly linking extent of occurrence, ecological specialization, and observed recovery following local extirpation. If recovery time exceeds the frequency of disturbance, local extirpation can lead to local extinction. Given that ENSO-induced disturbances are increasing in frequency, in severity, and in geographic scale, these results suggest that specialist species with restricted geographic ranges could be at particularly high risk of global extinction.

Diversity patterns in butterfly communities of the Greek nature reserve Dadia

Traditionally, nature reserves have been centered mainly around areas that are important for vertebrate diversity. This practice has not gone unchallenged and may be a suboptimal choice for overall conservation planning. To investigate this problem, we sampled butterfly species richness in a nature reserve in north eastern Greece that was originally established for the protection of birds of prey. Patterns of butterfly species richness and abundance were investigated by means of transect walks across a range of the seven predominant habitat types (wet and dry meadow, pine, oak and mixed forest, grazed, and agricultural land). Data analysis, including ANOVA and DCA (detrended correspondence analysis), revealed that the main gradients in butterfly species richness (low to high) were from sites dominated by the pine forest matrix of the core areas of the reserve, to peripheral sites in landscapes of mixed or oak forest, and from sites with little human impact to more disturbed areas with high grazing pressure. Species of conservation interest were concentrated at sites of low human impact. Ten of them are endemic to Europe and/or threatened in Europe. In this respect, the most important species are Lycaena ottomanus, Thymelicus acteon, and Pseudophilotes vicrama which are declining all over Europe. Our results suggest that (1) traditional agricultural practices in areas surrounded by forest can be considered as important management tools in butterfly conservation, (2) highest butterfly species richness is found in the periphery of the reserve rather than in the core areas, and (3) for butterfly conservation the zones surrounding the strictly protected areas are equally important as the core areas.

Butterfly, spider, and plant communities in different land-use types in Sardinia, Italy

Butterfly, spider, and plant species richness and diversity were investigated in five different land-use types in Sardinia. In 16 one-hectare plots we measured a set of 15 environmental variables to detect the most important factors determining patterns of variation in species richness, particularly endemicity. The studied land-use types encompassed homogeneous and heterogeneous shrublands, shrublands with tree-overstorey, Quercus forest and agricultural land. A total of 30 butterfly species, among which 10 endemics, and 50 spider (morpho)species, were recorded. Butterfly and spider community composition differed according to land-use type. The main environmental factors determining diversity patterns in butterflies were the presence of flowers and trees. Spiders reacted mainly to habitat heterogeneity and land-use type. Traditional land-use did not have adverse effects on the diversity of butterflies, spiders, or plants. The number of endemic butterfly species per treatment increased with total species richness and altitude. Butterfly and spider richness did not co-vary across the five land-use types. Butterflies were, however, positively associated with plant species richness and elevation, whereas spiders were not. Conclusively, butterflies did not appear to be good indicators for spider diversity and species richness at the studied sites.

Bacterial community composition and predicted functional ecology of sponges, sediment and seawater from the thousand islands reef complex, West Java, Indonesia

In the present study, we assessed the composition of Bacteria in four biotopes namely sediment, seawater and two sponge species (Stylissa massa and Xestospongia testudinaria) at four different reef sites in a coral reef ecosystem in West Java, Indonesia. In addition to this, we used a predictive metagenomic approach to estimate to what extent nitrogen metabolic pathways differed among bacterial communities from different biotopes. We observed marked differences in bacterial composition of the most abundant bacterial phyla, classes and orders among sponge species, water and sediment. Proteobacteria were by far the most abundant phylum in terms of both sequences and Operational Taxonomic Units (OTUs). Predicted counts for genes associated with the nitrogen metabolism suggested that several genes involved in the nitrogen cycle were enriched in sponge samples, including nosZ, nifD, nirK, norB and nrfA genes. Our data show that a combined barcoded pyrosequencing and predictive metagenomic approach can provide novel insights into the potential ecological functions of the microbial communities. Not only is this approach useful for our understanding of the vast microbial diversity found in sponges but also to understand the potential response of microbial communities to environmental change.