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      Enhancing African coelacanth monitoring using environmental DNA

      research-article
      1 , 2 , , 3 , , 3 , 4 , 5 , 2 , 2 , 6
      Biology Letters
      The Royal Society
      eDNA, coelacanth, marine biodiversity, DNA monitoring, management, South Africa

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          Abstract

          Coelacanths are rare, elusive, ancient lobe-finned fish species, residing in poorly accessible tropical marine caves and requiring close monitoring and protection. Environmental DNA (eDNA) approaches are being increasingly applied in the detection of rare and threatened species. Here we devise an eDNA approach to detect the presence of African coelacanths ( Latimeria chalumnae) off the eastern coast of South Africa. Novel coelacanth-specific primers were designed to avoid cross-amplification with other fish lineages and validated for specificity. These primers were tested on field samples in conjunction with remotely operated vehicle (ROV) visual surveys. Samples were collected from a known coelacanth habitat and two adjacent slope habitats a few kilometres apart. Coelacanth DNA was detected from three of 15 samples collected. Two of these positive eDNA detections occurred in the presence of coelacanths, as evidenced by ROV footage, while the third positive detection was at a station where coelacanths had not been previously observed. eDNA detections are discussed in relation to the species’ metabolic rate, movement patterns and population size, as well as the local oceanographic features. We demonstrate that eDNA can provide a non-invasive method to extend the knowledge of coelacanth distribution ranges and boost research efforts around these iconic fishes.

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          Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities.

          mothur aims to be a comprehensive software package that allows users to use a single piece of software to analyze community sequence data. It builds upon previous tools to provide a flexible and powerful software package for analyzing sequencing data. As a case study, we used mothur to trim, screen, and align sequences; calculate distances; assign sequences to operational taxonomic units; and describe the alpha and beta diversity of eight marine samples previously characterized by pyrosequencing of 16S rRNA gene fragments. This analysis of more than 222,000 sequences was completed in less than 2 h with a laptop computer.
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            VSEARCH: a versatile open source tool for metagenomics

            Background VSEARCH is an open source and free of charge multithreaded 64-bit tool for processing and preparing metagenomics, genomics and population genomics nucleotide sequence data. It is designed as an alternative to the widely used USEARCH tool (Edgar, 2010) for which the source code is not publicly available, algorithm details are only rudimentarily described, and only a memory-confined 32-bit version is freely available for academic use. Methods When searching nucleotide sequences, VSEARCH uses a fast heuristic based on words shared by the query and target sequences in order to quickly identify similar sequences, a similar strategy is probably used in USEARCH. VSEARCH then performs optimal global sequence alignment of the query against potential target sequences, using full dynamic programming instead of the seed-and-extend heuristic used by USEARCH. Pairwise alignments are computed in parallel using vectorisation and multiple threads. Results VSEARCH includes most commands for analysing nucleotide sequences available in USEARCH version 7 and several of those available in USEARCH version 8, including searching (exact or based on global alignment), clustering by similarity (using length pre-sorting, abundance pre-sorting or a user-defined order), chimera detection (reference-based or de novo), dereplication (full length or prefix), pairwise alignment, reverse complementation, sorting, and subsampling. VSEARCH also includes commands for FASTQ file processing, i.e., format detection, filtering, read quality statistics, and merging of paired reads. Furthermore, VSEARCH extends functionality with several new commands and improvements, including shuffling, rereplication, masking of low-complexity sequences with the well-known DUST algorithm, a choice among different similarity definitions, and FASTQ file format conversion. VSEARCH is here shown to be more accurate than USEARCH when performing searching, clustering, chimera detection and subsampling, while on a par with USEARCH for paired-ends read merging. VSEARCH is slower than USEARCH when performing clustering and chimera detection, but significantly faster when performing paired-end reads merging and dereplication. VSEARCH is available at https://github.com/torognes/vsearch under either the BSD 2-clause license or the GNU General Public License version 3.0. Discussion VSEARCH has been shown to be a fast, accurate and full-fledged alternative to USEARCH. A free and open-source versatile tool for sequence analysis is now available to the metagenomics community.
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              Primer3—new capabilities and interfaces

              Polymerase chain reaction (PCR) is a basic molecular biology technique with a multiplicity of uses, including deoxyribonucleic acid cloning and sequencing, functional analysis of genes, diagnosis of diseases, genotyping and discovery of genetic variants. Reliable primer design is crucial for successful PCR, and for over a decade, the open-source Primer3 software has been widely used for primer design, often in high-throughput genomics applications. It has also been incorporated into numerous publicly available software packages and web services. During this period, we have greatly expanded Primer3’s functionality. In this article, we describe Primer3’s current capabilities, emphasizing recent improvements. The most notable enhancements incorporate more accurate thermodynamic models in the primer design process, both to improve melting temperature prediction and to reduce the likelihood that primers will form hairpins or dimers. Additional enhancements include more precise control of primer placement—a change motivated partly by opportunities to use whole-genome sequences to improve primer specificity. We also added features to increase ease of use, including the ability to save and re-use parameter settings and the ability to require that individual primers not be used in more than one primer pair. We have made the core code more modular and provided cleaner programming interfaces to further ease integration with other software. These improvements position Primer3 for continued use with genome-scale data in the decade ahead.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: Writing – original draft
                Role: InvestigationRole: MethodologyRole: Writing – review and editing
                Role: ConceptualizationRole: MethodologyRole: SupervisionRole: ValidationRole: Writing – review and editing
                Role: ConceptualizationRole: Funding acquisitionRole: Project administrationRole: SupervisionRole: Writing – review and editing
                Role: MethodologyRole: Writing – review and editing
                Role: MethodologyRole: Project administrationRole: SupervisionRole: ValidationRole: Writing – review and editing
                Journal
                Biol Lett
                Biol Lett
                rsbl
                roybiolett
                Biology Letters
                The Royal Society
                1744-9561
                1744-957X
                October 2024
                October 23, 2024
                October 23, 2024
                : 20
                : 10
                : 20240415
                Affiliations
                [ 1 ] Department of Ichthyology and Fisheries Science, Rhodes University; , Makhanda, South Africa
                [ 2 ] South African Institute for Aquatic Biodiversity (SAIAB); , Makhanda, South Africa
                [ 3 ] School of Biological and Environmental Sciences, Liverpool John Moores University; , Liverpool L3 3AF, UK
                [ 4 ] South African National Biodiversity Institute (SANBI); , Cape Town, South Africa
                [ 5 ] Institute for Coastal and Marine Research, Nelson Mandela University; , Gqeberha (Port Elizabeth), South Africa
                [ 6 ] Department of Biochemistry and Microbiology, Rhodes University; , Makhanda, South Africa
                Author notes

                Electronic supplementary material is available online at https://doi.org/10.6084/m9.figshare.c.7477916.

                [ † ]

                Present address: South African Institute for Aquatic Biodiversity (SAIAB), Makhanda, South Africa

                Author information
                https://orcid.org/0000-0002-4227-1226
                https://orcid.org/0000-0001-8154-9828
                https://orcid.org/0000-0002-5329-0553
                https://orcid.org/0000-0001-5051-573X
                https://orcid.org/0009-0004-7157-3647
                https://orcid.org/0000-0001-6206-696X
                Article
                rsbl20240415
                10.1098/rsbl.2024.0415
                11496946
                39439356
                5820fceb-1fd8-4df7-865d-a41fc8ad08d2
                © 2024 The Author(s).

                Published by the Royal Society under the terms of the Creative Commons Attribution License http://creativecommons.org/licenses/by/4.0/, which permits unrestricted use, provided the original author and source are credited.

                History
                : July 18, 2024
                : September 10, 2024
                : September 17, 2024
                Funding
                Funded by: NRF Postgraduate Scholarship Programme (MND210406592334) to J.-C.O.;
                Funded by: UKRI Natural Environment Research Council grant NE/T007028/1 to P.S. and S.M.;
                Funded by: African Coelacanth Ecosystem (ACEP) under the DSI-NRF (grant 129216) to K.J.S.;
                Funded by: UK Research and Innovation through the Global Challenges Research Fund (NE/S008950/1) to J.-C.O. and K.J.S.;
                Categories
                1001
                60
                Marine Biology
                Research Articles

                Life sciences
                edna,coelacanth,marine biodiversity,dna monitoring,management,south africa
                Life sciences
                edna, coelacanth, marine biodiversity, dna monitoring, management, south africa

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