Cold-water corals are conspicuous in the waters off Eastern Canada. Despite that, there are few DNA sequence records from specimens collected in the region available in GenBank, and not all species recorded in the region have sequence data regardless of geographic origin. This can limit the use of eDNA techniques to detect and identify corals. Our objective was to sequence and publish sequences for two octocoral DNA barcoding markers: CO1 and MutS. We sequenced and deposited 36 sequences to GenBank from 19 specimens representing three sea pen taxa (Octocorallia: Pennatuloidea): Distichoptilum gracile, Pennatula aculeata, and Protoptilum carpenteri. Identification of all specimens was confirmed by B. M. Neves before submission. Specimens and DNA tissues were donated to the Canadian Museum of Nature, where they are currently stored. This publication is part 1 of a series of GenBank submissions by our lab. Specimens were collected from across the Northwest Atlantic and originate from depths ranging between 200-1924 meters. Specimens were collected as part of research vessel multispecies trawl surveys or remotely operated vehicle (ROV ROPOS) surveys. DNA was isolated and purified using the QIAgen DNeasy Blood and Tissue kit, with an initial overnight incubation with Proteinase K. Two commonly used octocoral barcoding regions were amplified using previously described primers: 1) COII8068F (McFadden et al., 2004) and COIOCTR (France and Hoover, 2002) for the CO1 gene, and 2) ND42599F (France and Hoover, 2002) and mut3458R (Sánchez et al., 2003) for the MutS gene. Amplifications were conducted using 12.5 µl of Green DreamTaq Master Mix (Thermo Fisher Scientific), 1 µl of template DNA, 0.5 µl of each 10 µM forward and reverse primers, 0.5 µl of 10 µM reverse primer, and 10.5 µl of water. Thermocycling was run as follows: 3 min of initial denaturation at 95 °C, followed by 40 cycles at 95 °C for 30 s, 30 s at annealing temperature of 48 °C, then 65 s at an extension temperature of 72 °C, and a final elongation at 72 °C for 4 min. PCR products were cleaned using Agencourt AMPure XP Beads (Beckman Coulter) and sent to The Center for Advanced Genomics, Toronto, Canada for Sanger sequencing. Sequences were visualized and aligned using Geneious Prime 2022.0.2. Obtained sequences have been deposited in GenBank under accession numbers OQ569768- OQ569784 and OQ420359- OQ420377. This work was funded by Fisheries and Oceans Canada under an Enhanced Regional Capacity grant (2020-2021) and the Marine Conservation Targets (MCT) program (2021-2024), Newfoundland and Labrador Region.

Fisheries and Oceans Canada’s (DFO) National Biofouling Monitoring Program (BMP) has conducted annual field surveys to monitor the introduction, establishment, spread, species richness, and relative abundance of native and non-indigenous species (NIS) since 2006. Standardized monitoring protocols employed by DFO-Maritimes, -Gulf, and -Quebec Regions include biofouling collector plates deployed from May to October at intertidal and shallow subtidal, geo-referenced sites, including public and private docks, aquaculture lease sites, public and private marinas and yacht clubs. Initially in the Maritimes Region (2006-2017) collectors consisted of 3, 10 cm by 10cm PVC plates deployed in a vertical array and spaced approximately 40-cm apart with the shallowest plate hung at least 1 m below the surface to sample shallow subtidal and intertidal species (Sephton et al. 2011, 2017). Two replicate arrays were deployed at least 5 m apart per site. Since 2018, collector arrays were modified to enhance statistical replication, including 10 individual collectors deployed per site at 1 m depth and at least 5 m apart (as above) from May to October. The percent cover of AIS on all collectors was determined by visual examination and scored as follows; (i) ‘0’ = absent, (ii) ‘1’ = ≤25 % cover, (iii) ‘2’ = 25 to ≤50 %, (iv) ‘3’ = 50–75% , and (v) ‘4’ = >75%. Average percent cover is provided for all NIS observed annually per site. Presence-absence indicates that an NIS was observed on at least one collector plate. One additional rocky intertidal species (Asian shore crab; Hemigrapsus sanguineus) was assessed via beach surveys as permitted by time and resources following its initial siting in St Mary’s Bay (Nova Scotia) in April 2020. Rapid assessment surveys conducted in the Fall of 2020 and 2021 were employed to delineate H. sanguineus’ distribution and relative abundance. Areas deemed suitable and at high risk for spread were targeted, including exposed rocky intertidal habitat in southwest regions of Nova Scotia and New Brunswick. Each rapid assessment consisted of 30-minute beach surveys per site conducted by 2 or 3 people (modified from Stephenson et al. 2011). During each survey, crabs were collected under rocks and seaweed in preferred cobble/boulder habitat (Lohrer et al. 2000). Count data was standardized for each site as the number of crabs collected per 30-min search per person. Cite as: DFO-Maritimes Biofouling Monitoring Program. Published October 2018, Updated December 2023. Coastal Ecosystems Science Division, Fisheries and Oceans Canada, Dartmouth, NS Citations: Sephton D, B Vercaemer, JM Nicolas, J Keays (2011) Monitoring for invasive tunicates in Nova Scotia, Canada (2006-2009) Aquatic Invasions 6: 391-403. Sephton D, B Vercaemer, A Silva, L Stiles, M Harris, K Godin (2017) Biofouling monitoring for aquatic invasive species (AIS) in DFO Maritimes Regions (Atlantic shore of Nova Scotia and southwest New Brunswick): May-November, 2012-2015. Canadian Technical Report of Fisheries and Aquatic Sciences 3158: 72 pp. Stephenson EH, RS Steneck, RH Seeley (2009) Possible temperature limits to range expansion of non-native Asian shore crabs in Maine. Journal of Experimental Marine Biology and Ecology 375: 21–31. doi:10.1016/j.jembe.2009.04.020