Community assembly and metaphylogeography of soil biodiversity: insights from haplotype-level community DNA metabarcoding within an oceanic island

Most of our understanding of island diversity comes from the study of aboveground systems, while the patterns and processes of diversification and community assembly for belowground biotas remain poorly understood. Here we take advantage of a relatively young and dynamic oceanic island to advance our understanding of eco-evolutionary processes driving community assembly within soil mesofauna. Using whole organism community DNA (wocDNA) metabarcoding and the recently developed metaMATE pipeline, we have generated spatially explicit and reliable haplotype-level DNA sequence data for soil mesofaunal assemblages sampled across the four main habitats within the island of Tenerife. Community ecological and metaphylogeographic analyses have been performed at multiple levels of genetic similarity, from haplotypes to species and supraspecific groupings. Broadly consistent patterns of local-scale species richness across different insular habitats have been found, whereas local insular richness is lower than in continental settings. Our results reveal an important role for niche conservatism as a driver of insular community assembly of soil mesofauna, with only limited evidence for habitat shifts promoting diversification. Furthermore, support is found for a fundamental role of habitat in the assembly of soil mesofauna, where habitat specialism is mainly due to colonisation and the establishment of preadapted species. Hierarchical patterns of distance decay at the community level and metaphylogeographical analyses support a pattern of geographic structuring over limited spatial scales, from the level of haplotypes through to species and lineages, as expected for taxa with strong dispersal limitations. Our results demonstrate the potential for wocDNA metabarcoding to advance our understanding of biodiversity.

Biological composition analysis of a natural medicine, Faeces Vespertilionis, with complex sources using DNA metabarcoding

Faeces Vespertilionis is a commonly used fecal traditional Chinese medicine. Traditionally, it is identified relying only on morphological characters. This poses a serious challenge to the composition analysis accuracy of this complex biological mixture. Thus, for quality control purposes, an accurate and effective method should be provided for taxonomic identification of Faeces Vespertilionis. In this study, 26 samples of Faeces Vespertilionis from ten provinces in China were tested using DNA metabarcoding. Seven operational taxonomic units (OTUs) were detected as belonging to bats. Among them, Hipposideros armiger (Hodgson, 1835) and Rhinolophus ferrumequinum (Schober and Grimmberger, 1997) were the main host sources of Faeces Vespertilionis samples, with average relative abundances of 59.3% and 24.1%, respectively. Biodiversity analysis showed that Diptera and Lepidoptera were the most frequently consumed insects. At the species level, 19 taxa were clearly identified. Overall, our study used DNA metabarcoding to analyze the biological composition of Faeces Vespertilionis, which provides a new idea for the quality control of this special traditional Chinese medicine.

One Bat’s Waste is Another Man’s Treasure: A DNA Metabarcoding Approach for the Assessment of Biodiversity and Ecosystem Services Using Bat Faeces

Arthropod populations are constantly changing due to changes in climate and the globalisation of trade and travel. Effective and diverse monitoring techniques are required to understand these changes. DNA metabarcoding has facilitated the development of a broad monitoring method to sample arthropod diversity from environmental and faecal samples. In this study, we applied DNA metabarcoding to DNA extracted from bat faecal pellets of Rhinolophus hipposideros, the lesser horseshoe bat in Ireland, a highly protected bat species of conservation concern in Europe. From as few as 24 bat faecal pellets, we detected 161 arthropod species, spanning 11 orders, including 38 pest species of which five were determined to be priority pests, highlighting important ecosystem services. We also report the identification 14 species not previously reported in Ireland, but upon further investigation found that many of these are likely misidentified due to inadequacies in the genetic reference database. For the first time, we were able to use non-invasively collected bat samples to examine the role of sex in the diet of bats and found that the male and female diets did not differ significantly. However, sampling location did explain variation within the diet, highlighting how landscape features influence arthropod composition and diversity. We discuss the current limitations of the methodology in Ireland, how these can be overcome in future studies, and how this data can be used for biodiversity monitoring and informing conservation management of protected bat species.

Idea paper: Monitoring and databasing non-native species to manage establishment debt in aquatic ecosystems

In the era of globalization, biological invasions are one of the most serious social issues. Thus, managing its impact is an urgent task. It is essential to control non-native species before they become established. However, it is insufficient to address establishment debt, which occurs when a non-native species has been introduced into an area but has not yet been established in the wild. In particular, unintentionally introduced or contaminated organisms of the aquatic ornamental pet trade are referred to as “hitchhikers” and have not received much attention in the context of establishment debt. To understand the nature of establishment debt, including that of aquatic hitchhikers, I propose the monitoring of non-native species inhabiting artificial isolated waters, such as indoor aquariums, and the construction of a database using environmental DNA metabarcoding. This idea would be an effective non-regulatory management approach when implemented broadly, at the country level. Furthermore, implementation of this strategy in combination with border biosecurity and field monitoring may promote accurate prioritization, rapid species identification, and effective invasion pathway assessment.

The protist community mirrors seasonality and mesoscale hydrographic features in the oligotrophic Sargasso Sea

Protists represent the majority of the eukaryotic diversity in the oceans. They have different functions in the marine food web, playing essential roles in the biogeochemical cycles. Meanwhile the available data is rich in horizontal and temporal coverage, little is known on their vertical structuring, particularly below the photic zone. The present study applies DNA metabarcoding to samples collected over three years in conjunction with the BATS time-series to assess marine protist communities in the epipelagic and mesopelagic zones. The protist community showed a dynamic seasonality in the epipelagic, responding to hydrographic yearly cycles. Mixotrophic lineages dominated throughout the year; however, autotrophs bloomed during the rapid transition between the winter mixing and the stratified summer, and heterotrophs had their peak at the end of summer, when the base of the thermocline reaches its deepest depth. Below the photic zone, the community, dominated by Rhizaria, is depth-stratified and relatively constant throughout the year, mirroring local hydrographic and biological features such as the oxygen minimum zone. The results suggest a dynamic partitioning of the water column, where the niche vertical position for each community changes throughout the year, likely depending on nutrient availability, the mixed layer depth, and other hydrographic features. Finally, the protist community closely followed mesoscale events (eddies), where the communities mirrored the hydrographic uplift, raising the deeper communities for hundreds of meters, and compressing the communities above.

Dealing with the promise of metabarcoding in mega-event biomonitoring: EXPO2015 unedited data

As human activities on our planet persist, causing widespread and irreversible environmental degradation, the need to biomonitor ecosystems has never been more pressing. These circumstances have required a renewal in monitoring techniques, encouraged by necessity to develop more rapid and accurate tools which will support timely observations of ecosystem structure and function. The World Exposition (from now 'EXPO2015') hosted in Milan from May to October 2015 was a global event that could be categorized as a mega-event, which can be defined as an acute environmental stressor, possibly generating biodiversity alteration and disturbance. During the six months of EXPO2015, exhibitors from more than 135 countries and 22 million visitors insisted on a 1.1 million square meters area. Faced with such a massive event, we explore the potential of DNA metabarcoding using three molecular markers to improve the understanding of anthropogenic impacts in the area, both considering air and water monitoring. Furthermore, we explore the effectiveness of the taxonomy assignment phase considering different taxonomic levels of analysis and the use of data mining approaches to predict sample origin. Unless the degree of taxa identification still remains open, our results showed that DNA metabarcoding is a powerful genomic-based tool to monitor biodiversity at the microscale, allowing us to capture exact fingerprints of specific event sites and to explore in a comprehensive manner the eukaryotic community alteration. With this work, we aim to disentangle and overcome the crucial issues related to the generalization of DNA metabarcoding in order to support future applications.

Inferring microbiota functions from taxonomic genes: a review

Deciphering microbiota functions is crucial to predict ecosystem sustainability in response to global change. High-throughput sequencing at the individual or community level has revolutionized our understanding of microbial ecology, leading to the big data era and improving our ability to link microbial diversity with microbial functions. Recent advances in bioinformatics have been key for developing functional prediction tools based on DNA metabarcoding data and using taxonomic gene information. This cheaper approach in every aspect serves as an alternative to shotgun sequencing. Although these tools are increasingly used by ecologists, an objective evaluation of their modularity, portability, and robustness is lacking. Here, we reviewed 100 scientific papers on functional inference and ecological trait assignment to rank the advantages, specificities, and drawbacks of these tools, using a scientific benchmarking. To date, inference tools have been mainly devoted to bacterial functions, and ecological trait assignment tools, to fungal functions. A major limitation is the lack of reference genomes—compared with the human microbiota—especially for complex ecosystems such as soils. Finally, we explore applied research prospects. These tools are promising and already provide relevant information on ecosystem functioning, but standardized indicators and corresponding repositories are still lacking that would enable them to be used for operational diagnosis.