Passive sampling of environmental DNA in aquatic environments using 3D-printed hydroxyapatite samplers

The study of environmental DNA released by aquatic organisms in their habitat offers a fast, non-invasive and sensitive approach to monitor their presence. Common eDNA sampling methods such as filtration and precipitation are time consuming, require human intervention and are not applicable to a wide range of habitats such as turbid waters and poorly-accessible environments. To circumvent these limitations, we propose to use the binding properties of minerals to create a passive eDNA sampler. We have designed 3D-printed samplers made of hydroxyapatite (HAp samplers), a mineral known for its high binding affinity with DNA. The shape and the geometry of the samplers have been designed to facilitate their handling in laboratory and field. Here we describe and test the ability of HAp samplers to recover artificial DNA and eDNA. We show that HAp samplers efficiently recover DNA and are effective even on small amounts of eDNA (<1 ng). However, we also observed large variations in the amount of DNA recovered even under controlled conditions. By better understanding the physico-chemical interactions between DNA and the HAp sampler surface, one could improve the repeatability of the sampling process and provide an easy-to-use eDNA sampling tool for aquatic environments.

[1]  Bessey Cindy,et al.  Passive eDNA collection enhances aquatic biodiversity analysis. , 2021, Communications biology.

[2]  O. Monnier,et al.  Exploring the capacity of aquatic biofilms to act as environmental DNA samplers: Test on macroinvertebrate communities in rivers. , 2020, The Science of the total environment.

[3]  Lauren C. Bergman,et al.  The need for robust qPCR‐based eDNA detection assays in environmental monitoring and species inventories , 2020, Environmental DNA.

[4]  Richard T. Corlett,et al.  Applications of environmental DNA (eDNA) in ecology and conservation: opportunities, challenges and prospects , 2020, Biodiversity and Conservation.

[5]  T. Minamoto,et al.  Estimating shedding and decay rates of environmental nuclear DNA with relation to water temperature and biomass , 2020, Environmental DNA.

[6]  L. Orlando,et al.  Unveiling the Ecological Applications of Ancient DNA From Mollusk Shells , 2020, Frontiers in Ecology and Evolution.

[7]  C. Richter,et al.  Reporting the limits of detection and quantification for environmental DNA assays , 2019, Environmental DNA.

[8]  G. Carvalho,et al.  Environmental DNA size sorting and degradation experiment indicates the state of Daphnia magna mitochondrial and nuclear eDNA is subcellular , 2019, Scientific Reports.

[9]  H. Doi,et al.  The detection of aquatic macroorganisms using environmental DNA analysis—A review of methods for collection, extraction, and detection , 2019, Environmental DNA.

[10]  S. Mariani,et al.  Sponges as natural environmental DNA samplers , 2019, Current Biology.

[11]  Klement Tockner,et al.  Emerging threats and persistent conservation challenges for freshwater biodiversity , 2018, Biological reviews of the Cambridge Philosophical Society.

[12]  Lynsey R. Harper,et al.  Limited dispersion and quick degradation of environmental DNA in fish ponds inferred by metabarcoding , 2018, bioRxiv.

[13]  J. Chevalier,et al.  Resorption of calcium phosphate materials: Considerations on the in vitro evaluation , 2018 .

[14]  Holly M. Bik,et al.  Acidity promotes degradation of multi-species environmental DNA in lotic mesocosms , 2018, Communications Biology.

[15]  Kristy Deiner,et al.  Environmental DNA metabarcoding: Transforming how we survey animal and plant communities , 2017, Molecular ecology.

[16]  A. Piaggio,et al.  Clearing muddied waters: Capture of environmental DNA from turbid waters , 2017, PloS one.

[17]  M. Lintermans,et al.  Methods to maximise recovery of environmental DNA from water samples , 2017, PloS one.

[18]  C. Gunsch,et al.  Adsorption capacity of multiple DNA sources to clay minerals and environmental soil matrices less than previously estimated. , 2017, Chemosphere.

[19]  S. Knudsen,et al.  Comparison of capture and storage methods for aqueous macrobial eDNA using an optimized extraction protocol: advantage of enclosed filter , 2017 .

[20]  L. Waits,et al.  Critical considerations for the application of environmental DNA methods to detect aquatic species , 2016 .

[21]  V. Kattimani,et al.  Hydroxyapatite—Past, Present, and Future in Bone Regeneration , 2016 .

[22]  M. P. Piggott Evaluating the effects of laboratory protocols on eDNA detection probability for an endangered freshwater fish , 2016, Ecology and evolution.

[23]  K. McKelvey,et al.  Environmental DNA particle size distribution from Brook Trout (Salvelinus fontinalis) , 2015, Conservation Genetics Resources.

[24]  L. Waits,et al.  Using environmental DNA methods to improve detectability in a hellbender (Cryptobranchus alleganiensis) monitoring program , 2015 .

[25]  C. Turner,et al.  Fish environmental DNA is more concentrated in aquatic sediments than surface water , 2015 .

[26]  Kristy Deiner,et al.  Special Issue Article: Environmental DNA Choice of capture and extraction methods affect detection of freshwater biodiversity from environmental DNA , 2015 .

[27]  M. Kondoh,et al.  The Release Rate of Environmental DNA from Juvenile and Adult Fish , 2014, PloS one.

[28]  C. Alemán,et al.  DNA adsorbed on hydroxyapatite surfaces. , 2014, Journal of materials chemistry. B.

[29]  F. Altermatt,et al.  Utility of Environmental DNA for Monitoring Rare and Indicator Macroinvertebrate Species , 2014, Freshwater Science.

[30]  B. Ludes,et al.  Adsorption of DNA on biomimetic apatites: Toward the understanding of the role of bone and tooth mineral on the preservation of ancient DNA , 2014 .

[31]  W. Yu,et al.  Adsorption of proteins and nucleic acids on clay minerals and their interactions: A review , 2013 .

[32]  Robert S. Arkle,et al.  Estimating occupancy and abundance of stream amphibians using environmental DNA from filtered water samples , 2013 .

[33]  M. Brundin,et al.  DNA binding to hydroxyapatite: a potential mechanism for preservation of microbial DNA. , 2013, Journal of endodontics.

[34]  V. Maheshwari,et al.  Adsorption and desorption of DNA on graphene oxide studied by fluorescently labeled oligonucleotides. , 2011, Langmuir : the ACS journal of surfaces and colloids.

[35]  M. Sakai,et al.  Effects of pH, ionic strength, and solutes on DNA adsorption by andosols , 2010, Biology and Fertility of Soils.

[36]  E. Crapster-Pregont,et al.  The adsorption of short single-stranded DNA oligomers to mineral surfaces. , 2011, Chemosphere.

[37]  Hadley Wickham,et al.  ggplot2 - Elegant Graphics for Data Analysis (2nd Edition) , 2017 .

[38]  J. Ascher,et al.  Extracellular DNA in soil and sediment: fate and ecological relevance , 2009, Biology and Fertility of Soils.

[39]  R. Cortès,et al.  DNA adsorption at liquid/solid interfaces. , 2008, The journal of physical chemistry. B.

[40]  J. Trevors,et al.  Cycling of extracellular DNA in the soil environment , 2007 .

[41]  Stephen L. R. Ellison,et al.  Routes to improving the reliability of low level DNA analysis using real-time PCR , 2006, BMC biotechnology.

[42]  R. Naiman,et al.  Freshwater biodiversity: importance, threats, status and conservation challenges , 2006, Biological reviews of the Cambridge Philosophical Society.

[43]  H. Chen,et al.  Adsorption of DNA on clay minerals and various colloidal particles from an Alfisol , 2006 .

[44]  A. Dorner-Reisel,et al.  Thermogravimetric and thermokinetic investigation of the dehydroxylation of a hydroxyapatite powder , 2004 .

[45]  M. Okazaki,et al.  Affinity binding phenomena of DNA onto apatite crystals. , 2001, Biomaterials.

[46]  M. Franchi,et al.  Clay-Nucleic Acid Complexes: Characteristics and Implications for the Preservation of Genetic Material in Primeval Habitats , 1999, Origins of life and evolution of the biosphere.

[47]  M. Khanna,et al.  X-ray diffractometry and electron microscopy of DNA from Bacillus subtilis bound on clay minerals , 1998 .

[48]  M. Khanna,et al.  Amplification of DNA bound on clay minerals , 1998 .

[49]  J. Harsh,et al.  Effects of DNA Polymer Length on Its Adsorption to Soils , 1994, Applied and environmental microbiology.

[50]  P. Simonet,et al.  Adsorption of DNA on clay minerals: protection against DNaseI and influence on gene transfer , 1992 .

[51]  M. Khanna,et al.  Transformation of Bacillus subtilis by DNA bound on montmorillonite and effect of DNase on the transforming ability of bound DNA , 1992, Applied and environmental microbiology.

[52]  J. G. Elias,et al.  The dimensions of DNA in solution. , 1981, Journal of molecular biology.

[53]  Senem Yetgin DNA adsorption on silica, alumina and hydroxyapatite and imaging of dna by atomic force microscopy , 2013 .

[54]  J. Lafon Synthèse, stabilité thermique et frittage d'hydroxyapatites carbonatées : , 2004 .

[55]  C. Gaillard,et al.  Avoiding adsorption of DNA to polypropylene tubes and denaturation of short DNA fragments. , 1998 .

[56]  D. Sparks,et al.  Kinetics of Ion Exchange on Clay Minerals and Soil: II. Elucidation of Rate-limiting Steps1 , 1986 .

[57]  C. Weber,et al.  Methods for measuring the acute toxicity of effluents to freshwater and marine organisms , 1985 .

[58]  J. Cortez,et al.  Reactions of nucleic acid bases with inorganic soil constituents , 1981 .