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      Low diversity of Angiostrongylus cantonensis complete mitochondrial DNA sequences from Australia, Hawaii, French Polynesia and the Canary Islands revealed using whole genome next-generation sequencing

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          Abstract

          Background

          Rats ( Rattus spp.) invaded most of the world as stowaways including some that carried the rat lungworm, Angiostrongylus cantonensis, the cause of eosinophilic meningoencephalitis in humans and other warm-blooded animals. A high genetic diversity of A. cantonensis based on short mitochondrial DNA regions is reported from Southeast Asia. However, the identity of invasive A. cantonensis is known for only a minority of countries. The affordability of next-generation sequencing for characterisation of A. cantonensis genomes should enable new insights into rat lung worm invasion and parasite identification in experimental studies.

          Methods

          Genomic DNA from morphologically verified A. cantonensis (two laboratory-maintained strains and two field isolates) was sequenced using low coverage whole genome sequencing. The complete mitochondrial genome was assembled and compared to published A. cantonensis and Angiostrongylus malaysiensis sequences. To determine if the commonly sequenced partial cox1 can unequivocally identify A. cantonensis genetic lineages, the diversity of cox1 was re-evaluated in the context of the publicly available cox1 sequences and the entire mitochondrial genomes. Published experimental studies available in Web of Science were systematically reviewed to reveal published identities of A. cantonensis used in experimental studies.

          Results

          New A. cantonensis mitochondrial genomes from Sydney (Australia), Hawaii (USA), Canary Islands (Spain) and Fatu Hiva (French Polynesia), were assembled from next-generation sequencing data. Comparison of A. cantonensis mitochondrial genomes from outside of Southeast Asia showed low genetic diversity (0.02–1.03%) within a single lineage of A. cantonensis. Both cox1 and cox2 were considered the preferred markers for A. cantonensis haplotype identification. Systematic review revealed that unequivocal A. cantonensis identification of strains used in experimental studies is hindered by absence of their genetic and geographical identity.

          Conclusions

          Low coverage whole genome sequencing provides data enabling standardised identification of A. cantonensis laboratory strains and field isolates. The phenotype of invasive A. cantonensis, such as the capacity to establish in new territories, has a strong genetic component, as the A. cantonensis found outside of the original endemic area are genetically uniform. It is imperative that the genotype of A. cantonensis strains maintained in laboratories and used in experimental studies is unequivocally characterised.

          Electronic supplementary material

          The online version of this article (10.1186/s13071-019-3491-y) contains supplementary material, which is available to authorized users.

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          Most cited references47

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          Reconstructing mitochondrial genomes directly from genomic next-generation sequencing reads—a baiting and iterative mapping approach

          We present an in silico approach for the reconstruction of complete mitochondrial genomes of non-model organisms directly from next-generation sequencing (NGS) data—mitochondrial baiting and iterative mapping (MITObim). The method is straightforward even if only (i) distantly related mitochondrial genomes or (ii) mitochondrial barcode sequences are available as starting-reference sequences or seeds, respectively. We demonstrate the efficiency of the approach in case studies using real NGS data sets of the two monogenean ectoparasites species Gyrodactylus thymalli and Gyrodactylus derjavinoides including their respective teleost hosts European grayling (Thymallus thymallus) and Rainbow trout (Oncorhynchus mykiss). MITObim appeared superior to existing tools in terms of accuracy, runtime and memory requirements and fully automatically recovered mitochondrial genomes exceeding 99.5% accuracy from total genomic DNA derived NGS data sets in <24 h using a standard desktop computer. The approach overcomes the limitations of traditional strategies for obtaining mitochondrial genomes for species with little or no mitochondrial sequence information at hand and represents a fast and highly efficient in silico alternative to laborious conventional strategies relying on initial long-range PCR. We furthermore demonstrate the applicability of MITObim for metagenomic/pooled data sets using simulated data. MITObim is an easy to use tool even for biologists with modest bioinformatics experience. The software is made available as open source pipeline under the MIT license at https://github.com/chrishah/MITObim.
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            Ecosystem Consequences of Biological Invasions

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              Biology and distribution of the rat lungworm, Angiostrongylus cantonensis, and its relationship to eosinophilic meningoencephalitis and other neurological disorders of man and animals.

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                Author and article information

                Contributors
                bara.cervena@gmail.com
                modryd@vfu.cz
                barborafeckova.vet@gmail.com
                kristyna@hrazdilova.cz
                pforonda@ull.edu.es
                amalonso@ull.edu.es
                Rogan.Lee@health.nsw.gov.au
                johndenisewalker@gmail.com
                niebuhrc@landcareresearch.co.nz
                richard.malik@sydney.edu.au
                jan.slapeta@sydney.edu.au
                Journal
                Parasit Vectors
                Parasit Vectors
                Parasites & Vectors
                BioMed Central (London )
                1756-3305
                16 May 2019
                16 May 2019
                2019
                : 12
                : 241
                Affiliations
                [1 ]ISNI 0000 0004 1936 834X, GRID grid.1013.3, Sydney School of Veterinary Science, , University of Sydney, ; Sydney, NSW 2006 Australia
                [2 ]ISNI 0000 0001 1009 2154, GRID grid.412968.0, Department of Pathology and Parasitology, , University of Veterinary and Pharmaceutical Sciences Brno, ; Palackého třída 1946/1, 612 42 Brno, Czech Republic
                [3 ]ISNI 0000 0001 1015 3316, GRID grid.418095.1, Institute of Vertebrate Biology, , Czech Academy of Sciences, ; Květná 8, 603 65 Brno, Czech Republic
                [4 ]Institute of Parasitology, Biology Center of the Czech Academy of Sciences, Branišovská 1160/31, 370 05 České Budějovice, Czech Republic
                [5 ]ISNI 0000 0001 1009 2154, GRID grid.412968.0, CEITEC VFU, University of Veterinary and Pharmaceutical Sciences Brno, ; Palackého třída 1946/1, 612 42 Brno, Czech Republic
                [6 ]ISNI 0000000121060879, GRID grid.10041.34, Instituto Universitario de Enfermedades Tropicales y Salud Pública de Canarias, , Universidad de La Laguna, ; C/Astrofisico F Sanchez, s/n, Tenerife, 38203 La Laguna, Canary Islands Spain
                [7 ]ISNI 0000000121060879, GRID grid.10041.34, Department Obstetricia y Ginecología, Pediatría, Medicina Preventiva y Salud Pública, Toxicología, Medicina Legal y Forense y Parasitología, , Universidad de La Laguna, ; 38203 San Cristóbal de La Laguna, Canary Islands Spain
                [8 ]ISNI 0000 0004 1936 834X, GRID grid.1013.3, Westmead Clinical School, , University of Sydney, ; Sydney, NSW 2145 Australia
                [9 ]ISNI 0000 0004 1936 834X, GRID grid.1013.3, Marie Bashir Institute for infectious Diseases and Biosecurity, , University of Sydney, ; Sydney, NSW 2006 Australia
                [10 ]USDA-APHIS-WS, National Wildlife Research Center, Hawaii Field Station, PO Box 10880, Hilo, HI 96721 USA
                [11 ]ISNI 0000 0004 1936 834X, GRID grid.1013.3, Centre for Veterinary Education, , University of Sydney, ; Sydney, NSW 2006 Australia
                [12 ]ISNI 0000 0001 0747 5306, GRID grid.419186.3, Present Address: Manaaki Whenua-Landcare Research, ; PO Box 69040, Lincoln, 7608 New Zealand
                Author information
                http://orcid.org/0000-0003-1459-9117
                Article
                3491
                10.1186/s13071-019-3491-y
                6524341
                31097040
                304296f2-ee34-412b-b490-31608783d3b3
                © The Author(s) 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 10 March 2019
                : 6 May 2019
                Funding
                Funded by: “Proyectos I + D de la Consejeria de Economía, Industria, Comercio y Conocimiento de la Comunidad Autónoma de Canarias” and FEDER 2014-2020
                Award ID: ID2017010092
                Award Recipient :
                Funded by: Red de Investigación de Centros de Enfermedades Tropicales-RICET, ISCIIISubdirección General de Redes y Centros de Investigación Cooperativa RETICS, Ministry of Health and Consumption, Spain
                Award ID: RD16/0027/0001
                Award Recipient :
                Funded by: International Collaboration in ecological and evolutionary biology of vertebrates
                Award ID: CZ02.2.69/0.0/0.0/16_027/0008027
                Award Recipient :
                Funded by: Internal Grant agency of UVPS
                Award ID: 123/2018/FVL
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100001823, Ministerstvo Školství, Mládeže a Tělovýchovy;
                Award ID: CEITEC 2020 (LQ1601)
                Award Recipient :
                Categories
                Research
                Custom metadata
                © The Author(s) 2019

                Parasitology
                rat lungworm,mitochondrial genome,genetic diversity,invasive species,next-generation sequencing,cox1,rattus

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