U.S Fish and Wildlife Services (USFWS) Lower Great Lakes Fish and Wildlife Conservation Office (LGLFWCO) Aquatic Invasive Species (AIS) Early Detection and Monitoring (EDM) program began in 2012 following the onset of Great Lakes Restoration Initiative (GLRI) funding. This program is apart of a larger basin-wide Great Lakes AIS EDM effort by the USFWS. The goal of this program is to detect novel AIS species (fish and aquatic invertebrates) by sampling a diversity of habitats with a wide array of gear types. Sampling focuses on harbors, rivers and tributaries of the lower Great Lakes (Erie and Ontario) as determined by a risk-based prioritization framework for AIS in the Great Lakes. The program is composed of three components of AIS surveillance: juvenile/adult fish, ichthyoplankton (larval fish), and aquatic invertebrates (including crayfish). Sampling strategies and protocols are analyzed and designed to capture the maximum species richness at locations with the assumption that capturing an abundance of species, including singletons and doubletons, could lead to detecting rare novel AIS species if present. Protocols vary depending on the component of the program and descriptions can be found within the metadata of each dataset. This program is adaptive in nature and standardization, although important for comparisons, is not the primary intention of this AIS sampling strategy. Any significant AIS detections are reported to partners following an internal communications protocol.

Benthic invertebrate surveys are a component of the U.S. Fish and Wildlife Services (USFWS) Alpena Fish and Wildlife Conservation Office (FWCO) Aquatic Invasive Species (AIS) Early Detection and Monitoring (EDM) Program. Benthic invertebrate surveys were included in the EDM program starting in 2014 with the goal of detecting novel AIS invertebrate species by sampling a diversity of habitats with a wide array of gear types. Surveys occurred annually from 2013 - 2024. Sampling focuses on harbors, rivers, and streams of Lake Huron and Lake Erie as determined by a risk-based prioritization framework for AIS in the Great Lakes. Benthic invertebrates are collected using a variety of sampling gears with the most utilized including: colonizers (rock bags and Hester-Dendy samplers) and sweep nets. The survey design targets the following three general taxon: Amphipoda, Bivalvia, and Gastropoda. These taxon are targeted due to the potential invasiveness and level of concern of species within those taxa as determined by Ecological Risk Screening Summaries (ERSS). The sampling season is typically between July and October - not all locations are sampled on a similar temporal scale, particularly those locations on the fringes of the programs surveillance range. Sampling strategies are analyzed and designed to capture the maximum species richness at locations with the assumption that capturing an abundance of species, including singletons and doubletons, could lead to detecting rare novel AIS species if present. Samples are preserved using 95% ethanol (EtOH) which allows for tissue preservation. The samples are then picked and sorted in-house (Alpena FWCO) by the three aforementioned general taxon. For several samples, these taxa groupings were screen for Region 3 watch list/target species. Some targeted sampling sites was sent to an external contractor for taxonomic identification a year or more later once funds became available. Given the dynamic and adaptive nature of AIS early detection, the list of watch list species has changed over time. The current watch list of invertebrates is as follows: Signal Crayfish, Spinycheek Crayfish, Yabby, Australian Redclaw Crayfish, Marbled Crayfish, Noble Crayfish, Red Swamp Crayfish, Harris Mud Crab, Bald Urchin Shrimp, Bloody Red Shrimp, Demon Shrimp, Killer Shrimp, Pontogammarus robustoides (no common name), Scud, Banded Mysterysnail, Chinese Mysterysnail, European Ear Snail, European Stream Valvata, Golden Mussel, Gravel Snail, Japanese Mysterysnail, Mud Bithynia (Faucet Snail), New Zealand Mudsnail, Quagga Mussel, and Zebra Mussel. Quagga and Zebra mussels are widespread in Lake Erie and Lake Huron and, with the exception of the St. Marys River, are no longer sorted or counted as bivalves because of the large number and frequency. The information within this dataset is geospatial in nature and documents benthic invertebrate sampling events. Both abiotic and biotic data is collected for each individual sampling event. It is possible that over time, the tools, gears, and instruments used to collect information have changed or been modified. The EDM Program is adaptive in nature and standardization, although important for comparisons, is not the primary intention of this AIS sampling strategy. Therefore, in some instances, the general statements made above regarding sample procedures may not apply. Also, identifying aquatic invertebrates such as amphipods, bivalves, and gastropods are difficult - especially when tasked with discovering novel AIS with limited support from reference materials (keys, voucher specimens, etc.). Because of this, some specimens collected cannot be assigned to the species level. Furthermore, due to these difficulties, identifications within this data set may be inaccurate and those records will remain within this data set unless otherwise detected and removed.

The U.S. Fish and Wildlife Services Great Lakes Aquatic Invasive Species (AIS) Early Detection and Monitoring (EDM) program began incorporating environmental DNA (eDNA) metabarcoding as a molecular AIS detection tool. From 2021-2023, the EDM metabarcoding program conducted pilot studies to determine the best approach for implementing a long-term multi-taxa geneomic monitoring program. Research objectives included the utility of using ethanol decant from ichthyoplankton (larval fish) samples for eDNA metabarcoding analysis along with a comparison against eDNA analysis from water filtration samples. The results from these years of experimental research has guided the EDM program to build a protocol around point water filtration eDNA samples. This multi-year eDNA project is the groundwork for what will become the EDM metabarcoding monitoring program at high priority AIS locations across the Great Lakes basin. The protocol and database for these slight variations of eDNA metabarcoding data collection are made available in this reference record. Data collected in 2021 and 2022 are in the same general format and within a single database. Data from 2023, because of the variation from 2021/2022, is in a separate database. Again, each year of these databases represents slightly different protocols or objectives that were determined prior to sample collection. However, the taxonomic assignment results underwent a similar pipeline each year. A metabarcoding eDNA detection means that a DNA sequence matching that species was assigned in the sample and does not necessarily mean that the fish was present at the time of sampling. Environmental DNA can enter the waterbody many ways including originating from live or dead fish, transported via boat, bird, or water current. For more information or questions, please contact the eDNA Program Coordinator, Nick Frohnauer at (nicholas_frohnauer@fws.gov).

Detection of environmental DNA (eDNA) has become a commonly used surveillance method for threatened or invasive vertebrates in both aquatic and terrestrial environments. However, use of eDNA methodologies for the detection of aquatic invertebrates (e.g., crayfish and insects) has been limited. Environmental DNA protocols can be especially useful for endangered invertebrates such as the Hine’s emerald dragonfly (Somatochlora hineana) where conservation efforts have been greatly hindered by the training, time, overall costs, and environmental impacts associated with conducting surveys in the calcareous fens occupied by this species. An essential step in developing such a protocol is to evaluate the dynamics of eDNA concentration under controlled and field conditions. In this study we examined the persistence and accumulation of eDNA from captive S. hineana larvae in experimental mesocosms at temperatures (5.0°C and 16.0°C) that reflect seasonal variation in their natural habitat, and we evaluated the usefulness of eDNA protocols for studying the distribution and abundance of invertebrates by assessing patterns of eDNA distribution for the Hine’s emerald dragonfly and its symbiont the devil crayfish, (Cambarus [=Lacunicambarus] diogenes) in the field over several months. In mesocosms, S. hineana eDNA persisted longer at 5.0°C but accumulated more readily at 16.0°C. In the field, life-history events affected seasonal variations in eDNA more significantly and consistently than temperature for both species. These data can be used to aid in conservation efforts for S. hineana and similar aquatic invertebrates.

Associated data for each of the three survey approaches (adult-juvenile, ichthyoplankton, eDNA) follow on individual tabs. Data rows correspond to individual sampling events. Distinctions are made below where survey approaches differ for the same column header. This dataset is associated with the following publication: Peterson, G., J. Hoffman, A. Trebitz, C. Hatzenbuhler, J. Myers, J. Ross, S. Okum, and E. Pilgrim. Early detection monitoring for non-indigenous fishes; comparison of survey approaches during two species introductions in a Great Lakes port. Biological Invasions. Springer SBM, New York, NY, USA, 463-478, (2022).

Associated data for each of the three survey approaches (adult-juvenile, ichthyoplankton, eDNA) follow on individual tabs. Data rows correspond to individual sampling events. Distinctions are made below where survey approaches differ for the same column header. This dataset is associated with the following publication: Peterson, G., J. Hoffman, A. Trebitz, C. Hatzenbuhler, J. Myers, J. Ross, S. Okum, and E. Pilgrim. Early detection monitoring for non-indigenous fishes; comparison of survey approaches during two species introductions in a Great Lakes port. Biological Invasions. Springer SBM, New York, NY, USA, 463-478, (2022).

PURPOSE: Provide early detection of newly arrived Aquatic Invasive Species (AIS) and determine the spread, establishment and spatial distribution of existing AIS within marine waters of the southern Gulf of St. Lawrence (sGSL), DFO Gulf Region boundaries (northern and eastern coastal shores of NB, Gulf shore of NS, and PEI shoreline). DESCRIPTION: DFO Science monitors for AIS in the Gulf Region. The data collected from DFO's biofouling monitoring program provides an overview of the distribution and abundance of Aquatic Invasive Species (AIS) in the Gulf Region. This information can be used by the general public, scientists and DFO managers. Monitoring program targeting aquatic invasive species (AIS). Native biofouling species are not included in this dataset. Botrylloides violaceus: Violet tunicate Botryllus schlosseri: Golden star tunicate Ciona intestinalis: Vase tunicate Styela clava: Clubbed tunicate Caprella mutica*: Japanese skeleton shrimp Membranipora membranacea: Coffin box bryozoan Carcinus maenas*: European green crab Codium fragile*: Oyster thief alga *indicates species that are not included as percent cover as they are not accurately captured by the sampling method, but are included as detections. Included here is a dataset of detection and percent cover data of AIS, as well as a monitoring station dataset. Environmental data collected, including from temperature loggers, are stored but not included here. PARAMETERS COLLECTED: Air and water temperature, salinity, depth, dissolved oxygen, weather conditions, list of biofouling AIS, percent cover of AIS on PVC plates. NOTES ON QUALITY CONTROL: Each sample and species is processed and identified in a standardized fashion using standardized DFO Science AIS protocols and taxonomic references. Data is manually entered into DFO Gulf Region's AIS Science biofouling database and randomly verified for accuracy. SAMPLING METHODS: Biofouling monitoring is conducted using PVC collector plates that are deployed in the water column approximately 1 meter below the sea surface in the spring of each year. Biofouling organisms settle on these plates which are collected in the fall of the same year. Abundances of AIS are given as percent plate cover. Physico-chemical data including temperature, conductivity, and depth as well as weather conditions are noted at each geo-referenced biofouling monitoring site during initial deployment and at time of retrieval. All biofouling organisms settled on the underside of the PVC plates are noted and percent cover of each AIS is estimated. USE LIMITATION: To ensure scientific integrity and appropriate use of the data, we would encourage you to contact the data custodian.

The Environmental Sample Processor (ESP), http://www.mbari.org/ESP/, is an autonomous biological sensing system that conducts in situ collection and molecular analysis of water samples and telemeters the results to shore in near real-time. The ESP can remotely detect and quantify abundance of target organisms using specific genetic probes. The probe generates a signal in the form of light, and an image of the array is taken using a camera and telemetered to shore for interpretation by experts. The intensity of the light signal is directly proportional to the abundance of the target that is present. Probes for 3 of the 4 primary HAB organisms in Puget Sound (i.e., Alexandrium, Heterosigma, and Pseudo-nitzschia) have already been used successfully on the ESP in the field. When deployed at key locations, the ESP can provide early warning of developing HABs and dramatically increase the opportunity for controlling the impacts of toxic blooms that can kill fish and contaminate shellfish. The goal of this project is to provide value added data to stakeholders in near real-time to improve early warning of HABs thereby reducing HAB-related economic losses and farmed-fish mortality and improving seafood safety. Another goal is to develop and test a method for use with the ESP to detect pathogenic Vibrio spp. (V. parahaemolyticus). Incorporating automated biosensor data into current risk and predictive models for the presence of HAB toxins and pathogens will result in a robust Health Early Warning System (HEWS). This work is designed to fill specific gaps in current risk and predictive models by providing rapid detection and reporting in real time for HABs and pathogens in conjunction with pertinent environmental data. The project will produce datasets describing the abundance for specific harmful algae and pathogenic bacteria at deployment locations in Puget Sound.

This dataset is associated with an examination of environmental DNA (eDNA) from the invasive round goby (Neogobius melanostomus) in parallel with visual surveys of fish count and size conducted by scuba divers in four Great Lakes: Michigan, Huron, Erie, and Ontario. Round goby inhabits benthic areas, and it is known to have direct contact with the bottom substrate as it feeds on dreissenid mussels. Considering this, eDNA samples were derived from multiple substrates: lake bottom water, sediment, and benthic algae; each manually collected by scuba divers from 3 or 6-meter depths and eight distinct transects. eDNA samples were analyzed by droplet digital PCR (ddPCR) and results are expressed as DNA Copy Numbers (CN) per 1 mL of water or per 1 gram of wet weight of sediment or benthic algae. All eDNA samples for this dataset were collected alongside a larger body of work conducted in 2022 (https://doi.org/10.5066/P13JDUMH) and relate to multiple years of work at these stations that includes: algal and dreissenid mussel biomass, water quality assessments, and diver observations of dreissenid mussels, round gobies, benthic substrate, and benthic algal cover. We refer to the benthic algae also as the 'Cladophora community' and 'submerged aquatic vegetation (SAV)' in other published project data, which were collected starting in 2018 (Great Lakes Science Center, 2018).