Detecting Snakehead Invaders. Natasha Serrao Robert Hanner Dirk Steinke

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1 Detecting Snakehead Invaders Natasha Serrao Robert Hanner Dirk Steinke

2 Family: Channidae Natasha Serrao Dirk Steinke Robert Hanner Legend- Native Distributions Parachanna distribution Channa distribution Courtenay and Williams 2004

3 Entry Pathways: 1) Ornamental fish 2) Live food fish C. hartcourtbutleri C. ornatipinnis C. species

4 Invasive populations in USA 1) C. argus 2) C. marulius 3) C. maculata

5 Purpose 1) Build a species-specific DNA sequence library for snakehead identification 2) Use the reference library to identify unknown invaders 3) Assess its utility by creating species-specific primer and probe sets for environmental DNA (edna) detection

6 DNA Barcoding DNA barcodes 652 bp of 5 region of cytochrome c oxidase subunit I (COI) mtdna (maternal inheritance) Effective for fish identification Supports identification at all life stages FISH-BOL global effort to create standardized library of fish DNA barcodes collaborators protocols; GPS, digital images, voucher specimens Data archived on BOLD; sequences submitted to Genbank

7 Determine identity of invasive species globally using library Species X (invasive population in Florida)

8 Distribution of snakehead specimens in their endemic and invaded regions Invaded/ Introduced Areas Endemic Range Native Range

9 DSCHA MCsn7 Channa_argus COI-5P NRSC NRS026 Channa_argus COI-5P NRSC NRS025 Channa_argus COI-5P ANGBF JQ JQ Channa_argus COI-5P NRSC NRS006 Channa_argus COI-5P DSCHA MC2sn1 Channa_argus COI-5P ANGBF JQ JQ Channa_argus COI-5P NRSC NRS050 Channa_argus COI-5P CYTC NC_ NC_ Channa_argus COI-5P DSCHA BCsnakehead5 Channa_sp. COI-5P DSCHA BCsnakehead4 Channa_sp. COI-5P DSCHA BCsnakehead6 Channa_sp. COI-5P DSCHA BCsnakehead7 Channa_sp. COI-5P NRSC NRS058 Channa_lucius COI-5P NRSC NRS060 Channa_lucius COI-5P NRSC NRS041 Channa_panaw COI-5P GBGC EU EU Channa_diplogramme COI-5P NRSC NRS044 Channa_diplogramme COI-5P NRSC NRS043 Channa_diplogramme COI-5P GBGC EU EU Channa_diplogramme COI-5P DSCHA CHAN021 Channa_micropeltes COI-5P DSCHA CHAN032 Channa_micropeltes COI-5P NRSC NRS030 Channa_micropeltes COI-5P NRSC NRS053 Channa_marulius COI-5P NRSC NRS001 Channa_marulius COI-5P NRSC NRS052 Channa_marulius COI-5P NRSC NRS048 Channa_marulius COI-5P ANGBF HM HM Channa_marulius COI-5P GBGC EU EU Channa_marulius COI-5P DSCHA CHAN040 Channa_asiatica COI-5P ANGBF FJ FJ Channa_punctata COI-5P GBGC EU EU Channa_punctata COI-5P ANGBF HM HM Channa_punctata COI-5P ANGBF HM HM Channa_punctata COI-5P ANGBF JN JN Channa_punctata COI-5P ANGBF HM HM Channa_striata COI-5P ANGBF HM HM Channa_striata COI-5P GBGC EU EU Channa_striata COI-5P ANGBF JQ JQ Channa_striata COI-5P ANGBF JQ JQ Channa_striata COI-5P NRSC NRS018 Channa_striata COI-5P BLB HQ Cstri4-LdB Channa_striata COI-5P BTL HQ Cstr5 Channa_striata COI-5P BTL HQ Cstr2 Channa_striata COI-5P BAFEN HM BNF_291 Parachanna_obscura COI-5P BAFEN HM BNF_290 Parachanna_obscura COI-5P PARO CHAN004b Parachanna_obscura COI-5P DSMIS OL-0736 Parachanna_africana COI-5P NRSC NRS037 Parachanna COI-5P NRSC NRS035 Parachanna_insignis COI-5P DSFRE OL-0220 Channa_pulchra COI-5P ANGBF FJ FJ Channa_orientalis COI-5P ANGBF FJ FJ Channa_orientalis COI-5P ANGBF JN JN Channa_orientalis COI-5P ANGBF JX JX Channa_orientalis COI-5P ANGBF HM HM Channa_bleheri COI-5P ANGBF HM HM Channa_bleheri COI-5P GBGC EU EU Channa_bleheri COI-5P ANGBF HM HM Channa_barca COI-5P DSCHA CHAN007 Channa_gachua COI-5P ANGBF HM HM Channa_gachua COI-5P ANGBF HM HM Channa_gachua COI-5P ANGBF HM HM Channa_stewartii COI-5P DSCHA CHAN026 Channa_stewartii COI-5P DSCHA CHAN008 Channa_stewartii COI-5P DSCHA CHAN041 Channa_sp. COI-5P ANGBF HM HM Channa_aurantimaculata COI-5P ANGBF HM HM Channa_aurantimaculata COI-5P ANGBF HM HM Channa_gachua COI-5P NRSC NRS012 Channa_gachua COI-5P NRSC NRS021 Channa_gachua COI-5P DSCHA CHAN034 Channa_gachua COI-5P PARO CHAN002 Channa_gachua COI-5P DSCHA CHAN012 Channa_gachua COI-5P DSCHA CHAN010 Channa_gachua COI-5P Neighbour-Joining Tree of Specimens 223 (total individuals); 116 BOLD; 107 Genbank represents 25/36 described species 0.02

10 C. gachua from Asia Phylogeographic variation Aids pathway anaylsis Myanmar & Thailand (17) Indonesia (2; 2) India (7; 6) 0.02

11 C. marulius from India and N. America high genetic diversity India (10; 7) N o rth Am erica (7) 0.02

12 C. argus invasives from United States Maryland New Jersey C. argus sourced from: VA, NY, PA Hybrid C. maculata(f)x C.argus(m) 0.02 C. argus C. argus NC_ C. sp. NRSC C. argus NRSC C. maculata NRSC No Voucher

13 Specimen NRSC027-11

14 Specimen NRSC re-identified as C. maculata (BC TNT market), 2011

15 Specimen NC_ (Wang and Yang 2011) potentially misidentified; no voucher specimen mention (Zhu et al. 2012) whole mitogenome sequences C. argus NC_ discordant

16 What about the BC specimen? Burnaby wild caught specimen Match to C. maculata and hybrid

17 Conclusions: Morphologically challenging to identify Taxonomy and barcode libraries remain incomplete Proper ID is critical to inform management decisions C. maculata C. argus (Herborg et al. 2007)

18 Next steps: qpcr for edna detection Species-specific primer and probe sets: P Barcode Region designed species-specific primer and probe sets for each target species based on barcode region probe sets were made for: C. argus, C. maculata, C. marulius

19 How will this affect us? high chance of colonization & establishment spawning and feeding conditions in Great Lakes favoured potential of spreading (Cudmore and Mandrak 2006)

20 Acknowledgements Universiti Sains Malaysia; Siti Azizah Mohd Nor, Jamsari A.M, Danial Jeffrey Eugene Hill (UFL) FWC; Kelly Gestring NYSDEC Bureau of Fisheries The Academy of Natural Sciences of Drexel University; Richard J. Horwitz and Paul Overbeck SFU; Jonathan Moore, Dave Scott, Corey Philis ENV; Matthias Herborg Nick Mandrak Walter Courtenay Chris Wilson Hanner Lab ROM; Erling Holm, Margaret Zur, Mary Burridge, Don Stacey, Hernán López- Fernández, Sean Insley DGIF; John Odenkirk UBC Eric Taylor FLMHN; Rob Robins, Larry Page, Terry Lott, Pamela S. Soltis NCSMNS; James Chang, Gabriela Hogue, Wayne Starnes CU; John Friel QU; Eleanor Adamson ZMUC; Tammes Menne University of Kerela; A. Biju Kumar

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