Metabarcoding the Global Fouling Community
Abstract
Temperate and tropical ports around the globe host a group of fouling animals and plants that are weeds tolerant of pollution, wit h short reproductive intervals and rapidly settling larvae or propagule stages. Most these organisms moved around the globe with shi pping and before biofouling science. Understanding fouling pressure from these dominant macrofoulers could be useful to management o f ship cleaning for example. We propose to use eDNA to quantify this common group of biofoulers. The initial work would be done in t he laboratories with water from off the docks in 3 geographically and temperature disparate harbors, one in Beaufort - NC, one in Po rt Sultan Qabbos; Muscat - Oman and one in Townsville - QLD , Australia. Weve identified 11 dominant fouling organisms in each harb or. In Beaufort, = Rittschof and Schultz will generate propagules in the laboratory, settle them, and extract their DNA/eDNA to ampl ify nuclear sequences for DNA barcoding. DNA extracted from individuals will be used to create a comprehensive database of sequences for identification of biofouling organisms. Initially, we will utilize qPCR to develop primers to amplify sequences from specific g roups of biofoulers (e.g. barnacles, tunicates, tubeworms, bryozoans etc.) and quality check our eDNA samples. Amplified products wi ll then be used to develop Target Sequence Capture (TSC) probes for next/3rd-generation sequencing (Illumina MiSeq and Nanopore) in order to selectively enhance for target organism DNA from the complex mixtures of sequences present in eDNA samples. We will optimiz e and standardize physical filtering process and time of collection and generate primers that specifically target our biofouling spe cies of interest. Our goal will be to develop a way to assess the macrofouling community and fouling pressure. Ideally we would deve lop a procedure that was qualitative and that could eventually be quantitative.If one examines the biofouling literature, especially published reports in peer review journals that originated from Office of Naval Research (ONR) programs, one readily identifies gr oups of biofoulers with cosmopolitan distributions from tropical to temperate regions (Rittschof, 2017, Rittschof and Holm 1997; Vas ishtha et al., 1995), Inter-site calibration (Holm et al, in prep) (Often the dominant biofoulers are the same species (e.g. Hydroid es elegans, Amphibalanus amphitrite, Bugula neritina), or closely related species with the same habits (eg. Hydroides spp., Bugula t urrita, Amphibalanus spp., colonial hydroids Obelia, Ectopleura; algae Enteromorpha spp.). Our plan is to develop an efficient water collection procedure and pipeline for sequence analyses of eDNA samples that can be deployed anywhere in the world to assess the pr esence and relative abundance of biofoulers. Examples of potential use are assessing biofouling in harbors, on stationary and moving sensors, in ballast water, and on ship hulls.In Phase 2, we would test our hypothesis of common fouling species by take the show o n the road and working with our colleagues that run the testing sites used for biofouling testing for the last 20 years in novel ma terials program (Singapore, Hawaii, Melbourne FL and San Louis Obispo) and having our colleagues assess the fouling communities desc ribed from eDNA in well studied biofouling communities. We anticipate this technology concept could be used with modification to wor k with fouling communities outside of harbors to assess fouling pressure in regions of navy interest.
Document Details
- Document Type
- DoD Grant Award
- Publication Date
- Oct 22, 2021
- Source ID
- N000142112965
Entities
People
- Madjid Delghandi
Organizations
- Office of Naval Research
- Sultan Qaboos University
- United States Navy