scholarly journals Metagenomic sequencing of dung beetle intestinal contents directly detects and identifies mammalian fauna

2016 ◽  
Author(s):  
Conrad P.D.T. Gillett ◽  
Andrew J. Johnson ◽  
Iain Barr ◽  
Jiri Hulcr
Keyword(s):  
High Throughput Sequencing ◽  
Sequence Data ◽  
Sampling Site ◽  
Dung Beetle ◽  
Dung Beetles ◽  
List Type ◽  
Metagenomic Sequencing ◽  
Biodiversity Assessment ◽  
Mammal Species ◽  
Intestinal Contents

SummaryCost, time, and expertise constraints limit traditional observation-based comprehensive biodiversity assessment. Therefore, surrogate focal taxa representative of wider biodiversity are commonly used as an imperfect ‘proxy’. Contemporary biodiversity assessments are also increasingly benefiting from the combination of high-throughput sequencing and metagenomic methodologies that enable identification of environmental DNA samples. However, there is a need for empirical studies combining the use of surrogate taxa with metagenomic approaches, that promise rapid and efficient biodiversity assessment.We here tested for the first time the possibility of using the intestinal contents of wild-collected dung beetles (Scarabaeidae) as a source of mammalian DNA, in a metagenomics proof-of-concept approach to directly detect and identify mammals from an area of savanna-scrub in southern Africa. Dung beetles have been purveyed as an indirect proxy measure of mammalian diversity, owing to their dependence upon vertebrate dung as a food source, and the ease with which they can be comprehensively sampled using simple and repeatable trapping protocols, achievable much faster than vertebrate surveys.Following shotgun sequencing of gut content DNA extractions from ten dung beetle species, we usedin silicofilters to identify mammals by searching the resulting reads against known mammalian mitochondrial DNA from online sequence repositories, matching 546 paired reads to known mitogenomes held in GenBank, and 634 reads to known mammal barcode sequences held in BOLD. Identified mammalian sequences were consistent with wild and domesticated ungulates known from the sampling site, and included blue wildebeest, plains zebra, and domestic cattle and goat. Four dung beetle samples yielded sufficient sequence data to successfully assemble the near-complete mitogenome of blue wildebeest at up to 21 X mean coverage, despite low initial DNA concentrations, unambiguously corroborating identification.It is conceptually and practically possible to rapidly and economically apply metagenomic techniques in dung beetle gut sequencing to detect the presence of mammals upon whose dung the beetles have fed. Since the approach can be readily scaled up, it may prove to be of practical use as a complement to traditional biodiversity assessment methods, and should be tested in usefulness for detecting rare, endangered or cryptic mammal species.

scholarly journals Dung beetles as vertebrate samplers – a test of high throughput analysis of dung beetle iDNA

2021 ◽  
Author(s):  
Rosie Drinkwater ◽  
Elizabeth L. Clare ◽  
Arthur Y. C. Chung ◽  
Stephen J. Rossiter ◽  
Eleanor M. Slade
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Dung Beetle ◽  
Blood Feeding ◽  
Preliminary Evidence ◽  
Dung Beetles ◽  
High Throughput Analysis ◽  
Humid Forest ◽  
Terrestrial Vertebrates ◽  
Wide Range

AbstractThe application of environmental DNA (eDNA) sampling in biodiversity surveys has gained widespread acceptance, especially in aquatic systems where free eDNA can be readily collected by filtering water. In terrestrial systems, eDNA-based approaches for assaying vertebrate biodiversity have tended to rely on blood-feeding invertebrates, including leeches and mosquitoes (termed invertebrate-derived DNA or iDNA). However, a key limitation of using blood-feeding taxa as samplers is that they are difficult to trap, and, in the case of leeches, are highly restricted to humid forest ecosystems. Dung beetles (superfamily Scarabaeoidea) feed on the faecal matter of terrestrial vertebrates and offer several potential benefits over blood-feeding invertebrates as samplers of vertebrate DNA. Importantly, these beetles can be easily captured in large numbers using simple, inexpensive baited traps; are globally distributed; and also occur in a wide range of biomes, allowing mammal diversity to be compared across habitats. In this exploratory study, we test the potential utility of dung beetles as vertebrate samplers by sequencing the mammal DNA contained within their guts. First, using a controlled feeding experiment, we show that mammalian DNA can be retrieved from the guts of large dung beetles (Catharsius renaudpauliani) for up to 10 hours after feeding. Second, by combining high-throughput sequencing of a multi-species assemblage of dung beetles with PCR replicates, we show that multiple mammal taxa can be identified with high confidence. By providing preliminary evidence that dung beetles can be used as a source of mammal DNA, our study highlights the potential for this widespread group to be used in future biodiversity monitoring surveys.

scholarly journals Assessment of metagenomic sequencing and qPCR for detection of influenza D virus in bovine respiratory tract samples

2020 ◽  
Author(s):  
Maodong Zhang ◽  
Yanyun Huang ◽  
Dale L. Godson ◽  
Champika Fernando ◽  
Trevor W. Alexander ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
High Throughput Sequencing ◽  
Sequence Data ◽  
Metagenomic Sequencing ◽  
Nanopore Sequencing ◽  
Sequence Detection ◽  
Nasal Swabs ◽  
Sequencing Platforms ◽  
Species Specific

AbstractHigh throughput sequencing is currently revolutionizing the genomics field and providing new approaches to the detection and characterization of microorganisms. The objective of this study was to assess the detection of influenza D virus (IDV) in bovine respiratory tract samples using two sequencing platforms (MiSeq and Nanopore (GridION)), and species-specific qPCR. An IDV-specific qPCR was performed on 232 samples (116 nasal swabs and 116 tracheal washes) that had been previously subject to virome sequencing using MiSeq. Nanopore sequencing was performed on 19 samples positive for IDV by either MiSeq or qPCR. Nanopore sequence data was analyzed by two bioinformatics methods: What’s In My Pot (WIMP, on the EPI2ME platform), and an in-house developed analysis pipeline. The agreement of IDV detection between qPCR and MiSeq was 82.3%, between qPCR and Nanopore was 57.9% (in-house) and 84.2% (WIMP), and between MiSeq and Nanopore was 89.5% (in-house) and 73.7% (WIMP). IDV was detected by MiSeq in 14 of 17 IDV qPCR-positive samples with Cq (cycle quantification) values below 31, despite multiplexing 50 samples for sequencing. When qPCR was regarded as the gold standard, the sensitivity and specificity of MiSeq sequence detection were 28.3% and 98.9%, respectively. We conclude that both MiSeq and Nanopore sequencing are capable of detecting IDV in clinical specimens with a range of Cq values. Sensitivity may be further improved by optimizing sequence data analysis, improving virus enrichment, or reducing the degree of multiplexing.

scholarly journals Assessment of Metagenomic Sequencing and qPCR for Detection of Influenza D Virus in Bovine Respiratory Tract Samples

Viruses ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 814 ◽  
Author(s):  
Maodong Zhang ◽  
Yanyun Huang ◽  
Dale L. Godson ◽  
Champika Fernando ◽  
Trevor W. Alexander ◽  
...  
Keyword(s):  
Respiratory Tract ◽  
High Throughput Sequencing ◽  
Sequence Data ◽  
Metagenomic Sequencing ◽  
Nanopore Sequencing ◽  
Sequence Detection ◽  
Nasal Swabs ◽  
Sequencing Platforms ◽  
Species Specific

High throughput sequencing is currently revolutionizing the genomics field and providing new approaches to the detection and characterization of microorganisms. The objective of this study was to assess the detection of influenza D virus (IDV) in bovine respiratory tract samples using two sequencing platforms (MiSeq and Nanopore (GridION)), and species-specific qPCR. An IDV-specific qPCR was performed on 232 samples (116 nasal swabs and 116 tracheal washes) that had been previously subject to virome sequencing using MiSeq. Nanopore sequencing was performed on 19 samples positive for IDV by either MiSeq or qPCR. Nanopore sequence data was analyzed by two bioinformatics methods: What’s In My Pot (WIMP, on the EPI2ME platform), and an in-house developed analysis pipeline. The agreement of IDV detection between qPCR and MiSeq was 82.3%, between qPCR and Nanopore was 57.9% (in-house) and 84.2% (WIMP), and between MiSeq and Nanopore was 89.5% (in-house) and 73.7% (WIMP). IDV was detected by MiSeq in 14 of 17 IDV qPCR-positive samples with Cq (cycle quantification) values below 31, despite multiplexing 50 samples for sequencing. When qPCR was regarded as the gold standard, the sensitivity and specificity of MiSeq sequence detection were 28.3% and 98.9%, respectively. We conclude that both MiSeq and Nanopore sequencing are capable of detecting IDV in clinical specimens with a range of Cq values. Sensitivity may be further improved by optimizing sequence data analysis, improving virus enrichment, or reducing the degree of multiplexing.

Seasonality, dung specificity and competition in dung beetle assemblages in the Australian Wet Tropics, north-eastern Australia

2005 ◽  
Vol 21 (1) ◽  
pp. 1-8 ◽  
Author(s):  
Karl Vernes ◽  
Lisa C. Pope ◽  
Christopher J. Hill ◽  
Felix Bärlocher
Keyword(s):  
Null Model ◽  
Dung Beetle ◽  
Dung Beetles ◽  
Maximum Temperature ◽  
Wet Season ◽  
Habitat Types ◽  
Mammal Species ◽  
Eastern Australia ◽  
Species Effect ◽  
Wet Tropics

A trapping study of five mammal species in wet sclerophyll forest adjacent to rain forest in the Australian Wet Tropics was used to examine the seasonal diversity, abundance and dung-specificity of dung beetles associated with mammal dung. A total of 542 dung beetles from 11 species within three genera was recovered from beneath the traps of 1104 mammal captures. The diversity of beetles associated with the dung of the northern bettong (Bettongia tropica), a mycophagous marsupial, differed significantly from the diversity predicted by a null model. Numbers of beetles varied significantly with type of dung, indicating preference by beetles. Beetle numbers were related positively to a 1-mo lag in monthly mean minimum temperature and less strongly to maximum temperature and rainfall. Significantly more beetles per mammal capture were detected in the wet season than in the dry season. Dung beetles showed a strong preference for either the Eucalyptus woodland (six species) or the adjacent Allocasuarina forest (four species), with only one species occurring in both habitat types. Beetle species from the Eucalyptus woodland were typically only detected in the late wet and early dry seasons, while species in the wetter Allocasuarina forest were generally collected during the late dry and early wet seasons. A significant ‘checkerboard’ species effect was detected in both time and space in both habitat types, suggesting that competition for dung was strong.

scholarly journals Dung beetle community assemblages in a southern African landscape: niche overlap between domestic and wild herbivore dung

2021 ◽  
pp. 1-12
Author(s):  
B. Sands ◽  
N. Mgidiswa ◽  
S. Curson ◽  
C. Nyamukondiwa ◽  
R. Wall
Keyword(s):  
Niche Overlap ◽  
Dung Beetle ◽  
Dung Beetles ◽  
Sub Saharan Africa ◽  
Cattle Dung ◽  
Mammal Species ◽  
Wide Range ◽  
Domestic Livestock ◽  
Abundance And Diversity ◽  
Diurnal Species

Abstract Dung beetles provide important ecosystem functions in semiarid environments, improving the physiochemical characteristics of the soil through tunnelling and burying nutrient-rich dung. In sub-Saharan Africa, diverse indigenous mammal communities support highly abundant dung beetle populations in savannah ecosystems. However, the conversion of landscapes to livestock agriculture may result in changes in the abundance and diversity of wild mammal species. This is likely to have significant impacts on dung beetle communities, particularly because domestic livestock dung may be contaminated with toxic residues of veterinary parasiticides. The environmental impact is likely to be affected by the degree of niche overlap between the beetle communities that colonize cattle dung and those that colonize the dung of wild mammals. We compared dung beetle communities between a pristine national park habitat dominated by large wild herbivores, and a pastoral farming community dominated by domestic livestock. Diurnal dung beetles were attracted to cattle dung in greater abundance and diversity compared to elephant, zebra or giraffe dung. Nocturnal/crepuscular dung beetles were attracted to non-ruminant dung (elephant and zebra) in higher abundance compared to ruminant dung (cattle and giraffe). Although there were no clear trophic specializations, three diurnal species showed an association with cattle dung, whereas eight nocturnal/crepuscular species showed an association with non-ruminant (elephant and zebra) dung. Diurnal species may be at greater risk from the toxic effects of residues of veterinary parasiticides in domestic livestock dung. Although many species showed trophic associations with wild herbivore dung, these beetles can utilize a wide range of dung and will readily colonize cattle dung in the absence of other options. As more land is converted to livestock agriculture, the contamination of dung with toxic residues from veterinary parasiticides could therefore negatively impact the majority of dung beetle species.

scholarly journals Dung beetle–mammal associations: methods, research trends and future directions

2019 ◽  
Vol 286 (1897) ◽  
pp. 20182002 ◽  
Author(s):  
Elizabeth H. Raine ◽  
Eleanor M. Slade
Keyword(s):  
Ecosystem Functioning ◽  
Environmental Gradients ◽  
Association Studies ◽  
Terrestrial Ecosystems ◽  
Dung Beetle ◽  
Field Studies ◽  
Dung Beetles ◽  
Mammal Species ◽  
Wide Range ◽  
Species Richness And Abundance

Dung beetles are increasingly used as a study taxon—both as bioindicators of environmental change, and as a model system for exploring ecosystem functioning. The advantages of this focal taxon approach are many; dung beetles are abundant in a wide range of terrestrial ecosystems, speciose, straightforward to sample, respond to environmental gradients and can be easily manipulated to explore species-functioning relationships. However, there remain large gaps in our understanding of the relationship between dung beetles and the mammals they rely on for dung. Here we review the literature, showing that despite an increase in the study of dung beetles linked to ecosystem functioning and to habitat and land use change, there has been little research into their associations with mammals. We summarize the methods and findings from dung beetle–mammal association studies to date, revealing that although empirical field studies of dung beetles rarely include mammal data, those that do, indicate mammal species presence and composition has a large impact on dung beetle species richness and abundance. We then review the methods used to carry out diet preference and ecosystem functioning studies, finding that despite the assumption that dung beetles are generalist feeders, there are few quantitative studies that directly address this. Together this suggests that conclusions about the effects of habitat change on dung beetles are based on incomplete knowledge. We provide recommendations for future work to identify the importance of considering mammal data for dung beetle distributions, composition and their contributions to ecosystem functioning; a critical step if dung beetles are to be used as a reliable bioindicator taxon.

scholarly journals Microbe Finder (MiFi®): Implementation of an Interactive Pathogen Detection Tool in Metagenomic Sequence Data

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 250
Author(s):  
Andres S. Espindola ◽  
Kitty F. Cardwell
Keyword(s):  
High Throughput ◽  
Web Application ◽  
Pathogen Detection ◽  
High Throughput Sequencing ◽  
Sequence Data ◽  
Metagenomic Data ◽  
Diagnostic Tools ◽  
Metagenomic Sequencing ◽  
Metagenomic Sequence ◽  
Metagenomic Sequence Data

Agricultural high throughput diagnostics need to be fast, accurate and have multiplexing capacity. Metagenomic sequencing is being widely evaluated for plant and animal diagnostics. Bioinformatic analysis of metagenomic sequence data has been a bottleneck for diagnostic analysis due to the size of the data files. Most available tools for analyzing high-throughput sequencing (HTS) data require that the user have computer coding skills and access to high-performance computing. To overcome constraints to most sequencing-based diagnostic pipelines today, we have developed Microbe Finder (MiFi®). MiFi® is a web application for quick detection and identification of known pathogen species/strains in raw, unassembled HTS metagenomic data. HTS-based diagnostic tools developed through MiFi® must pass rigorous validation, which is outlined in this manuscript. MiFi® allows researchers to collaborate in the development and validation of HTS-based diagnostic assays using MiProbe™, a platform used for developing pathogen-specific e-probes. Validated e-probes are made available to diagnosticians through MiDetect™. Here we describe the e-probe development, curation and validation process of MiFi® using grapevine pathogens as a model system. MiFi® can be used with any pathosystem and HTS platform after e-probes have been validated.

scholarly journals Wood Pastures: A Transitional Habitat between Forests and Pastures for Dung Beetle Assemblages

Forests ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 25
Author(s):  
László Somay ◽  
Viktor Szigeti ◽  
Gergely Boros ◽  
Réka Ádám ◽  
András Báldi
Keyword(s):  
Functional Group ◽  
Animal Health ◽  
Habitat Type ◽  
Dung Beetle ◽  
Dung Beetles ◽  
Livestock Grazing ◽  
Pitfall Traps ◽  
Assemblage Composition ◽  
Beetle Assemblages

Wood pastures are home to a variety of species, including the dung beetle. Dung beetles are an important functional group in decomposition. Specifically, in terms of livestock manure, they not only contribute to nutrient cycling but are key players in supporting human and animal health. Dung beetles, however, are declining in population, and urgent recommendations are needed to reverse this trend. Recommendations need to be based on solid evidence and specific habitats. Herein, we aimed to investigate the role of an intermediate habitat type between forests and pastures. Wood pastures are key areas for dung beetle conservation. For this reason, we compared dung beetle assemblages among forests, wood pastures, and grasslands. We complemented this with studies on the effects of dung type and season at three Hungarian locations. Pitfall traps baited with cattle, sheep, or horse dung were used in forests, wood pastures, and pasture habitats in spring, summer, and autumn. Dung beetle assemblages of wood pastures showed transient characteristics between forests and pastures regarding their abundance, species richness, Shannon diversity, assemblage composition, and indicator species. We identified a strong effect of season and a weak of dung type. Assemblage composition proved to be the most sensitive measure of differences among habitats. The conservation of dung beetles, and the decomposition services they provide, need continuous livestock grazing to provide fresh dung, as well as the maintenance of wood pastures where dung beetle assemblages typical of forests and pastures can both survive.

scholarly journals 673 Precision microbiome mapping identifies a microbiome signature predictive of Immune checkpoint inhibitor response across multiple research study cohorts

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A711-A711
Author(s):  
Matthew Robinson ◽  
Kevin Vervier ◽  
Simon Harris ◽  
David Adams ◽  
Doreen Milne ◽  
...  
Keyword(s):  
Gut Microbiome ◽  
Immune Checkpoint ◽  
Immune Checkpoint Inhibitor ◽  
Checkpoint Inhibitor ◽  
Advanced Melanoma ◽  
List Type ◽  
Metagenomic Sequencing ◽  
Genome Database ◽  
Sequencing Platform ◽  
Melanoma Patients

BackgroundThe gut microbiome of cancer patients appears to be associated with response to Immune Checkpoint Inhibitor (ICIs) treatment.1–4 However, the bacteria linked to response differ between published studies.MethodsLongitudinal stool samples were collected from 69 patients with advanced melanoma receiving approved ICIs in the Cambridge (UK) MELRESIST study. Pretreatment samples were analysed by Microbiotica, using shotgun metagenomic sequencing. Microbiotica’s sequencing platform comprises the world’s leading Reference Genome Database and advanced Microbiome Bioinformatics to give the most comprehensive and precise mapping of the gut microbiome. This has enabled us to identify gut bacteria associated with ICI response missed using public reference genomes. Published microbiome studies in advanced melanoma,1–3renal cell carcinoma (RCC) and non-small cell lung cancer (NSCLC)4 were reanalysed with the same platform.ResultsAnalysis of the MELRESIST samples showed an overall change in the microbiome composition between advanced melanoma patients and a panel of healthy donor samples, but not between patients who subsequently responded or did not respond to ICIs. However, we did identify a discrete microbiome signature which correlated with response. This signature predicted response with an accuracy of 93% in the MELRESIST cohort, but was less predictive in the published melanoma cohorts.1–3 Therefore, we developed a bioinformatic analytical model, incorporating an interactive random forest model and the MELRESIST dataset, to identify a microbiome signature which was consistent across all published melanoma studies. This model was validated three times by accurately predicting the outcome of an independent cohort. A final microbiome signature was defined using the validated model on MELRESIST and the three published melanoma cohorts. This was very accurate at predicting response in all four studies combined (91%), or individually (82–100%). This signature was also predictive of response in a NSCLC study and to a lesser extent in RCC. The core of this signature is nine bacteria significantly increased in abundance in responders.ConclusionsAnalysis of the MELRESIST study samples, precision microbiome profiling by the Microbiotica Platform and a validated bioinformatic analysis, have enabled us to identify a unique microbiome signature predictive of response to ICI therapy in four independent melanoma studies. This removes the challenge to the field of different bacteria apparently being associated with response in different studies, and could represent a new microbiome biomarker with clinical application. Nine core bacteria may be driving response and hold potential for co-therapy with ICIs.Ethics ApprovalThe study was approved by Newcastle & North Tyneside 2 Research Ethics Committee, approval number 11/NE/0312.ReferencesMatson V, Fessler J, Bao R, et al. The commensal microbiome is associated with anti-PD-1 efficacy in metastatic melanoma patients. Science 2018;359(6371):104–108.Gopalakrishnan V, Spencer CN, Nezi L, et al. Gut microbiome modulates response to anti-PD-1 immunotherapy in melanoma patients. Science 2018;359(6371):97–103.Frankel AE, Coughlin LA, Kim J, et al. Metagenomic shotgun sequencing and unbiased metabolomic profiling identify specific human gut microbiota and metabolites associated with immune checkpoint therapy efficacy in melanoma patients. Neoplasia 2017;19(10):848–855.Routy B, Le Chatelier E, Derosa L, et al. Gut microbiome influences efficacy of PD-1-based immunotherapy against epithelial tumors. Science 2018;359(6371):91–97.

scholarly journals MetaDEGalaxy: Galaxy workflow for differential abundance analysis of 16s metagenomic data

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 726
Author(s):  
Mike W.C. Thang ◽  
Xin-Yi Chua ◽  
Gareth Price ◽  
Dominique Gorse ◽  
Matt A. Field
Keyword(s):  
Microbial Communities ◽  
Sequence Data ◽  
Metagenomic Data ◽  
Marker Genes ◽  
Metagenomic Sequencing ◽  
Differential Analysis ◽  
Biomedical Sciences ◽  
Metagenomic Sequence ◽  
Differential Abundance ◽  
Differential Abundance Analysis

Metagenomic sequencing is an increasingly common tool in environmental and biomedical sciences.  While software for detailing the composition of microbial communities using 16S rRNA marker genes is relatively mature, increasingly researchers are interested in identifying changes exhibited within microbial communities under differing environmental conditions. In order to gain maximum value from metagenomic sequence data we must improve the existing analysis environment by providing accessible and scalable computational workflows able to generate reproducible results. Here we describe a complete end-to-end open-source metagenomics workflow running within Galaxy for 16S differential abundance analysis. The workflow accepts 454 or Illumina sequence data (either overlapping or non-overlapping paired end reads) and outputs lists of the operational taxonomic unit (OTUs) exhibiting the greatest change under differing conditions. A range of analysis steps and graphing options are available giving users a high-level of control over their data and analyses. Additionally, users are able to input complex sample-specific metadata information which can be incorporated into differential analysis and used for grouping / colouring within graphs.  Detailed tutorials containing sample data and existing workflows are available for three different input types: overlapping and non-overlapping read pairs as well as for pre-generated Biological Observation Matrix (BIOM) files. Using the Galaxy platform we developed MetaDEGalaxy, a complete metagenomics differential abundance analysis workflow. MetaDEGalaxy is designed for bench scientists working with 16S data who are interested in comparative metagenomics.  MetaDEGalaxy builds on momentum within the wider Galaxy metagenomics community with the hope that more tools will be added as existing methods mature.

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