Wednesday, May 2, 2018

You Are So Transparent (And That’s a Good Thing)

From politics to Facebook, transparency is on all our minds. And science, of course, depends on transparency to advance.

A big barrier to transparency in scientific research is the cost of viewing peer reviewed articles. A simple Google® Scholar search on a topic of your choosing will result in thousands of articles, many of which you might want to read. Unfortunately, to review an article requires payment of $30 or more. Who can afford that?

The mission of open access sites like PLOS-Public Library of Science (
http://journals.plos.org/) is to breach the cost barrier and promote the free flow of peer reviewed research.

Granted, the focus of PLOS seems to be on medical research. And yet, even so, an article by ExxonMobil researchers Prosset and Hedgpeth, is a part of the trove of treasures in this collection.

Viva transparency!

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Effects of bioturbation on environmental DNA migration through soil media
Christopher M. Prosser & Bryan M. Hedgpeth
PLOS  Published: April 24, 2018 • https://doi.org/10.1371/journal.pone.0196430
Contributed equally to this work with: Christopher M. Prosser, Bryan M. Hedgpeth
Roles: Conceptualization, Methodology, Writing
* E-mail: Christopher.m.prosser@exxonmobil.com
ORCID: http://orcid.org/0000-0002-5209-1878  Bryan M. Hedgpeth
Affiliation: ExxonMobil Biomedical Sciences Incorporated, Annandale, NJ, United States of America  
Abstract
Extracting and identifying genetic material from environmental media (i.e. water and soil) presents a unique opportunity for researchers to assess biotic diversity and ecosystem health with increased speed and decreased cost as compared to traditional methods (e.g. trapping). The heterogeneity of soil mineralogy, spatial and temporal variations however present unique challenges to sampling and interpreting results. Specifically, fate/transport of genetic material in the terrestrial environment represents a substantial data gap. Here we investigate to what degree, benthic fauna transport genetic material through soil. Using the red worm (Eisenia fetida), we investigate how natural movement through artificial soil affect the transport of genetic material. All experiments were run in Frabill® Habitat® II worm systems with approximately 5 cm depth of artificial soil. We selected an ªexoticº source of DNA not expected to be present in soil, zebrafish (Danio rerio) tissue. Experiment groups contained homogenized zebrafish tissue placed in a defined location combined with a varying number of worms (10, 30 or 50 worms per experimental group). Experimental groups comprised two controls and three treatment groups (representing different worm biomass) in triplicate. A total of 210 soil samples were randomly collected over the course of 15 days to investigate the degree of genetic transfer, and the rate of detection. Positive detections were identified in 14% - 38% of samples across treatment groups, with an overall detection rate of 25%. These findings highlight two important issues when utilizing environmental DNA for biologic assessments. First, benthic fauna are capable of redistributing genetic material through a soil matrix. Second, despite a defined sample container and abundance of worm biomass, as many as 86% of the samples were negative. This has substantial implications for researchers and managers who wish to interpret environmental DNA results from terrestrial systems. Studies such as these will aid in future study protocol design and sample collection methodology. Introduction Accurate biodiversity assessments are a central component to compliance with environmental regulations. In the United States, for example, environmental impact statements under the National Environmental Policy Act require extensive baseline information on biodiversity. Similarly, quantitative biodiversity assessments are important for assessing the progress of habitat reclamation efforts. However, traditional biodiversity monitoring relies on direct (ex. traps, sightings) or indirect (ex. tracks, calls) observation of organisms. Especially true for direct methods such as trapping or netting, these activities are often time consuming, expensive, and impractical in remote or hard to reach regions. Over the past decade, technological advances have resulted in the ability to detect the presence of organisms through amplification of environmental DNA (eDNA). eDNA is a generic term collectively referring to all genetic material that can be extracted from environmental media. Examples are extracellular DNA fragments, hair, feces, blood, free microbial cells, pollen or any other source by which cells and/or tissue may enter the environment [1]. Due to high precision, species-specific detection and low rates of false positives, eDNA has been increasingly utilized for an array of studies including biodiversity assessments, mapping of species distributions, and detection of invasive and endangered species [1±5]. DNA-based ecosystem monitoring can have distinct advantages over traditional sampling methods, including being less invasive/less destructive than trapping/netting. Sampling only environmental media (water, soil, sediment), reduces stress and danger of entrapment of valuable (e.g. endangered) species in nets or snares. The DNA sequencing of bulk material containing the DNA of dozens or hundreds of species would have been cost-prohibitive with older low throughput DNA sequencing platforms (e.g. Sanger sequencing). However, with next generation DNA sequencers (NGS), which use high-throughput technologies such as massively parallel sequencing, it is now possible to generate millions of DNA reads from bulk material in a short period of time [6]. Additionally, newer DNA sequencing technologies boast low detection limits (10−8 ng/μL) allowing for low levels of genetic material to be amplified and sequenced. To date, the majority of eDNA studies have focused on aquatic and/or wetland systems [3, 7±9]. This is most likely due to methodological advantages of sampling aquatic media. For example, lotic and lentic systems provide defined boundaries within which to sample and relatively large volumes of water (as large as several liters) can be filtered to concentrate available genetic material. In contrast to eDNA analysis from aquatic/marine systems, there is generally a paucity of data from terrestrial habitats. Soil matrices present unique challenges that are not encountered in aquatic systems. For example, the volume of soil used in extractions is typically a limiting factor (~0.25g soil per extraction). Additionally, little is known on eDNA fate and mobility in terrestrial systems over time and space (i.e. once deposited, there is little data to predict transport and/or persistence). A non-detect may be a false-negative if in fact the complexity of soil matrix precludes homogenous distribution of genetic material thus limiting spatial area from which it can be detected. As compared to aqueous media, the chemical complexity and reactivity of soils displays a greater degree of spatial and temporal heterogeneity, raising questions about eDNA mobility in soils. Soil mineralogy (e.g. clay, sand, silt) and subsequent mixtures (e.g. silty clays) will greatly influence the amount of surface reactive particles present, and thus the adsorption of genetic material within that matrix [10]. Physicochemical interactions influencing eDNA mobility within the soil matrix are highly variable and will depend on DNA fragment size, soil mineralogy, hydrophobicity, pH and ionic strength [11]. Persistence of eDNA in soils has also received limited attention and is incompletely understood. The presence of clay and soil colloids has been suggested to prohibit enzymatic degradation of genetic material thus potentially prolonging its availability for detection [12,13]. In anoxic environments, such as lake sediment, eDNA has been recovered dating back thousands of years [14]. Conversely, eDNA can also be taken up by bacteria as a source of nutrition expediting its removal from the environment [10]. Such uncertainties have led to wide estimates in persistence ranging from days to years in the top 15 cm of soil [15]. To date, few field studies have been conducted specifically focused on eDNA extraction from soil. However, in recent years researchers have investigated soil samples from natural wetland habitats [16] as well as in more spatially defined zoos and parks [17]. Fahner et al. [16] investigated large-scale plant monitoring using DNA metabarcoding. Researchers collected core samples from the Ramsar designated Peace-Athabasca Delta in Wood Buffalo National Park, Alberta, Canada with the goal of identifying standard DNA markers designed to evaluate floral biodiversity. An important approach in this study was the targeting of full length amplicons (400±900 base pairs), demonstrating this length is not so extensively degraded to preclude their use in biodiversity assessment. Andersen et al. [17] investigated a fundamental relationship between known species abundance and detectable levels of eDNA. Researchers isolated and amplified eDNA from known species in safari parks, zoological gardens, and farms and found that detectable eDNA generally reflected the diversity of animals on the landscape. However, these researchers reported patchy detection (as low as 31%) from soil surface. Researchers also reported eDNA extraction efficiency was inversely proportional to organic carbon content of the soil. The vast majority of studies to date have focused on the presence/absence of DNA in the environment; however, such studies do little to investigate eDNA fate and transport. While there are some exceptions in aquatic systems (i.e. [8]), there is a noticeable data gap investigating such effects in terrestrial systems. While the deposition of genetic material through normal processes (e.g. hair loss) is generally accepted, the degree to which physical (e.g. wind/rain) and biological (bioturbation) processes disseminate genetic material through terrestrial media are not well understood. As eDNA continues to grow as a tool for use in ecological assessments, a fundamental understanding of detection rate, and the risk of false negatives in terrestrial media will bolster data interpretation. Given the paucity of data related to eDNA fate and transport within terrestrial environments, the scope of this study focused on whether bioturbation will transport eDNA through soil. Our study was designed to investigate if normal biotic activity (e.g. the natural movement of worms through soil) would transport detectable levels of genetic material from a single, well defined depositional source, to adjacent areas. The redworm (Eisenia fetida) was used in controlled laboratory experiments to examine if, and to what degree bioturbation moves DNA from a single deposition source through soil. 
source:
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0196430
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Here are some excerpts from the PLOS Web site …

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Founded in 2001 as an alternative to the growing constraints of traditional scientific publishing, the Public Library of Science (PLOS) rapidly evolved into a driving force in the Open Access movement.
source:
https://www.plos.org/history
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TIP: Search the PLOS Web (http://journals.plos.org/plosone/) using whatever term you are interested in … like, for example, desulfurization. Then read to your heart’s content.


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