Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta Pellucida) in the Grand River, Ontario, 2019

Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta pellucida) in the Grand River, Ontario, 2019

Robin C. Gáspárdy and D. Andrew R. Drake

Ontario and Prairie Region Fisheries and Oceans Canada Great Lakes Laboratory for Fisheries and Aquatic Sciences 867 Lakeshore Road Burlington, ON L7S 1A1

2021

Canadian Data Report of Fisheries and Aquatic Sciences 1322

Canadian Data Report of Fisheries and Aquatic Sciences

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Canadian Data Report of Fisheries and Aquatic Sciences 1322

2021

Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta pellucida) in the Grand River, Ontario, 2019

Robin C. Gáspárdy and D. Andrew R. Drake

Ontario and Prairie Region Great Lakes Laboratory for Fisheries and Aquatic Sciences Fisheries and Oceans Canada 867 Lakeshore Road Burlington, ON L7S 1A1

Correct citation for this publication: Gáspárdy, R.C. and Drake, D.A.R. 2021. Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta pellucida) in the Grand River, Ontario, 2019. Can. Data Rep. Fish. Aquat. Sci. 1322: vii + 24 p.

TABLE OF CONTENTS

ABSTRACT ...... vi

RÉSUMÉ ...... vii

INTRODUCTION ...... 1

METHODS ...... 1 Study System and Site Selection ...... 1 Fish Assemblage Sampling ...... 1 Tagging ...... 3 Habitat Sampling ...... 3 Sampling Permits and Data Archiving ...... 3

RESULTS ...... 4 Fish Assemblage Sampling ...... 4 Tagging ...... 4 Habitat Sampling ...... 5

ACKNOWLEDGEMENTS ...... 6

REFERENCES ...... 6

iii

LIST OF FIGURES Figure 1. Detections of Eastern Sand Darter in the Grand River by DFO for the period 2004-2018 by seine net and benthic trawling. All sampling in 2019 between May and November was conducted at the 2019 study site at Cockshutt Bridge...... 7 Figure 2. Detections of Eastern Sand Darter by DFO 2004-2018 at Cockshutt Bridge (Cockshutt Rd. and Erie Ave., Brantford), representing 15% of DFO’s total Eastern Sand Darter detections in the Grand River (n = 291 Eastern Sand Darter in 14 sampling events)...... 8 Figure 3. Photograph of Eastern Sand Darter (Ammocrypta pellucida) captured by seining in June 2019. Orange VIE tag on left of anal fin visible through translucent body...... 9 Figure 4. Field sketches indicating area sampled by seine on a) July 16, 2019, b) October 2, 2019, and by trawl on c) October 28, 2019. The large riffle in the collection site is indicated by four wavy lines, shown perpendicular to river flow...... 9 Figure 5. Relationship between total length (mm) and mass (g) of Eastern Sand Darter captured between May and November in the Grand River, Ontario, 2019 (n = 304)...... 10 Figure 6. Length-frequency of Eastern Sand Darter (ESD) captured in the Grand River, Ontario between May and November, 2019 (n = 304). Overall, the mean total length (mm) of all Eastern Sand Darter captured was 62.69 (range: 46 – 72 mm)...... 10 Figure 7. Length-frequency of Eastern Sand Darter (ESD) captured in each sampling period between May and November, 2019 in the Grand River, Ontario. Mean total length (TL) provided below each panel...... 11 Figure 8. Photographs of VIE tagged Eastern Sand Darter recaptured a) once in another sampling period (first capture in August, recaptured in September), and b) twice in two other sampling periods (first capture in July, recaptured in August and September)...... 12 Figure 9. Semi-hourly river discharge (m3/s) sensor data from ECCC Hydrometric Station (02GB001) on the Grand River at Brantford, approximately 5.5 km upstream of 2019 sampling site. Sampling periods are highlighted, indicating dates and times during which sampling for Eastern Sand Darter was conducted at Cockshutt Bridge...... 13 LIST OF TABLES Table 1. Summary of sampling periods, effort, gear type, and number of Eastern Sand Darter captured. Effort is displayed as number of hauls or tows for each gear type, seine net (SN) and benthic trawl (TRL), respectively...... 14 Table 2. Summary of total length (mm) and mass (g) of Eastern Sand Darter captured by sampling period...... 14 Table 3. Summary of mean abiotic habitat values over each sampling period. Mean river discharge calculated from semi-hourly discharge sensor data extracted from Environment and Climate Change Canada real-time hydrometric monitoring station website...... 15 LIST OF APPENDICES Appendix 1. Visible Implant Elastomer (VIE) tag colour and location codes for Eastern Sand Darter assessed in physiology experiments by sampling period...... 16 Appendix 2. Sampling date, field code, sampling duration, gear type, and fishing effort for captures of Eastern Sand Darter (ESD) in the Grand River, 2019, to support physiology experiments. Where gear code SN = Seine (bag seine, 9.14 x1.8 m, 3 mm mesh), and TRL = Trawl (Gerken Siamese Trawl, 2.4 m foot rope, 3 mm outer cod mesh)...... 16 iv

Appendix 3. Individual total length (TL; mm), mass (M; g), and Visual Implant Elastomer (VIE) tag and recapture data for Eastern Sand Darter captured at Cockshutt Bridge, Grand River, 2019. Whether the fish was released, kept, tagged that day, and/or recaptured (RECAP) are noted (Y = yes, N = no). All fish were tagged on the ventral surface to the left or right of the anal fin. Tag codes refer to VIE tag colour and position in relation to the anal fin. Recapture tag code(s) lists all existing tags on the individual. Individuals recaptured within the same sampling period as tagged (*) were not re- tagged with the same code...... 17 Appendix 4. Summary of field notes indicating detections of other darter species captured during targeted sampling for Eastern Sand Darter in the Grand River, Ontario, 2019, where 1 = detected, 0 = not detected. Shown are dates where detection of other darter species was documented...... 23 Appendix 5. Daily abiotic habitat assessment measurements, collected immediately after sampling using a YSI EXO2 multiparameter sonde. Mean river discharge during sampling data calculated by averaging semi-hourly sensor data extracted from Environment and Climate Change Canada Real- time Hydrometric Data website on November 6, 2020...... 24

ABSTRACT Gáspárdy, R.C. and Drake, D.A.R. 2021. Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta pellucida) in the Grand River, Ontario, 2019. Can. Data Rep. Fish. Aquat. Sci. 1322: vii + 24 p. Targeted sampling for Eastern Sand Darter (Ammocrypta pellucida), a species listed as Threatened under Canada’s Species at Risk Act, was undertaken in the Grand River, Ontario during 2019. Because relatively little information exists about the physiological tolerances of Eastern Sand Darter, sampling was conducted to obtain individual fish for streamside experiments to evaluate the thermal and dissolved oxygen requirements of the species, in partnership with the University of Waterloo. The results of thermal and dissolved oxygen experiments will be disseminated in future publications; this data report summarizes field collections of Eastern Sand Darter near Brantford to undertake the experiments over the course of eight sampling periods between May and November, 2019. Seining was conducted in wadeable habitat (water depth < 1.2 m) between June and late October and benthic trawling was conducted in non-wadeable habitat (water depth > 1.2 m) in late October and November. Overall, 521 Easter Sand Darter were captured ranging from 48 to 72 mm total length; seining surveys (28 sampling days, 187 total hauls) resulted in the capture of 455 individuals, and trawling surveys (4 sampling days, 61 total tows) resulted in the capture of 66 individuals. A total of 282 Eastern Sand Darter were tagged with Visible Implant Elastomer (VIE) tags and 37 individuals were recaptured throughout sampling, including individuals recaptured across multiple sampling periods and nine individuals captured with both seining and trawling.

RÉSUMÉ Gáspárdy, R.C. and Drake, D.A.R. 2021. Summary of Targeted Sampling for Eastern Sand Darter (Ammocrypta pellucida) in the Grand River, Ontario, 2019. Can. Data Rep. Fish. Aquat. Sci. 1322: vii + 24 p. En 2019, on a mené des activités d’échantillonnage ciblant le dard de sable (Ammocrypta pellucida), une espèce menacée en vertu de la Loi sur les espèces en péril du Canada, dans la rivière Grand, en Ontario. Étant donné qu’il existe relativement peu de renseignements sur les seuils de tolérance physiologique de l’espèce, on a mené ces activités pour capturer des dards de sable en vue de mener des expériences en milieux riverains. Réalisées en collaboration avec l’Université de Waterloo, ces expériences visaient à évaluer les besoins du dard de sable en matière de température et d’oxygène dissous. Les résultats de ces expériences seront présentés dans des publications à venir. Le présent rapport de données résume les captures de dards de sable qui ont été réalisées sur le terrain, à proximité de Brantford, pendant huit périodes d’échantillonnage, de mai à novembre 2019, en vue de mener les expériences. On a effectué des relevés à la senne dans des milieux accessibles à gué (profondeur inférieure à 1,2 m) entre juin et la fin d’octobre, et des relevés au chalut dans des milieux non accessibles à gué (profondeur supérieure à 1,2 m) à la fin d’octobre et en novembre. Dans l’ensemble, on a capturé 521 dards de sable d’une longueur totale variant de 48 à 72 mm; 455 individus ont été capturés dans le cadre des relevés à la senne (28 jours de relevé et 187 calées) et 66 individus, dans le cadre des relevés au chalut (4 jours d’échantillonnage et 61 traits). Au total, on a marqué 282 dards de sable avec des étiquettes en élastomère visibles et on a recapturé 37 individus lors des activités d’échantillonnage. Parmi ceux-ci, certains ont été recapturés lors de différentes périodes d’échantillonnage et 9 individus ont été capturés à la fois lors des relevés à la senne et des relevés au chalut.

vii

INTRODUCTION Fisheries and Oceans Canada (DFO) has the responsibility to provide for the protection and recovery of fishes listed under the Species at Risk Act (SARA) of 2002. To inform scientific aspects of the recovery process, DFO regularly conducts field sampling to satisfy various research objectives for SARA-listed fishes, such as evaluating the distribution and abundance of species, determining species-habitat relationships, and better understanding the influence of threats and recovery actions. DFO data reports are published to support the Species at Risk Program by providing an overview of field activities and to provide a medium for archiving data associated with sampling SARA-listed fishes and their habitat. Eastern Sand Darter (Ammocrypta pellucida) is a small, elongate, translucent darter in the family Percidae. The Ontario designatable unit is listed as Threatened under SARA owing to its small and fragmented distribution and sensitivity to turbidity, contamination, and invasive species [Committee on the Status of Endangered Wildlife in Canada (COSEWIC) 2009; DFO 2011]. Like most SARA-listed freshwater fishes, relatively little information exists about the physiological tolerances of Eastern Sand Darter. To address this knowledge gap, streamside experiments were conducted in the Grand River near Brantford, Ontario to evaluate the thermal and dissolved oxygen requirements of the species, in partnership with the University of Waterloo. The results of thermal and dissolved oxygen experiments will be disseminated in future publications; this data report summarizes field collections of Eastern Sand Darter in the Grand River used for the physiological experiments over the course of eight visits between May and November, 2019.

METHODS

STUDY SYSTEM AND SITE SELECTION Eastern Sand Darter has been detected frequently by DFO within the Grand River between 2004 and 2018 (Figure 1), from an upstream limit near Brantford to a downstream limit near Cayuga (DFO unpublished data). All collections of Eastern Sand Darter in 2019 to support physiological experiments were conducted at Cockshutt Bridge, Erie Avenue, Brantford (access point coordinates: 43.11008, -80.24499), which is at the upstream end of a river section referred to locally as ‘the oxbow’. The preferred substrate of the species is clean sand and fine gravel (Dextrase et al. 2014) which is found in abundance at the Cockshutt Bridge access point. Extensive field sampling by DFO in this location has indicated a high likelihood of capturing Eastern Sand Darter; 15% of all Eastern Sand Darter detected by DFO in the Grand River between 2004 and 2018 were captured at this location by seining and trawling (DFO unpublished data; Figure 2). The site contains a public boat ramp and parking lot operated by the City of Brantford, which provided operational benefits for field sampling and bankside experiments.

FISH ASSEMBLAGE SAMPLING A targeted, opportunistic sampling approach was conducted. The purpose of sampling was to capture the number of Eastern Sand Darter needed for daily physiology experiments (maximum 20) while minimizing sampling effort; therefore, there was no set methodology in terms of prescribed effort or a standardized sampling approach. Sampling effort concluded daily when the required number of Eastern Sand Darter was reached. Each sampling period consisted of

3 – 5 sampling days within the same week, repeated every 3 – 5 weeks between May and November, 2019 (Table 1). Seining A 9.14 m long x 1.80 m tall bag seine with 3 mm mesh was used to collect Eastern Sand Darter within nearshore, wadeable (< 1.2 m in depth) habitat. Areas with slow river velocity and a dominance of sand, gravel, or silt substrate were targeted. Two to three field personnel pulled the seine in a downstream direction over a distance of approximately 30 to 50 m until Eastern Sand Darter was detected. Following initial detections, repeated hauls over shorter distances (10 to 20 m) were conducted in the vicinity of initial detections until the target number of Eastern Sand Darter was captured for the day. The total number of hauls per day was recorded. Benthic Trawling Seining failed to produce the target number of Eastern Sand Darter during late October, so benthic trawling was incorporated to allow sampling of areas with water depth greater than 1.2 m. As benthic trawling requires more equipment and places greater physical stress on captured fishes, the method was only used when seining was no longer effective in the late October and November period. The use of the benthic trawl also coincided with a number of large rainfall events resulting in increased water levels and unsafe wading conditions. A Gerken Siamese trawl was deployed at water depths between 1 and 3.5 m by hand from the front of a 5 m aluminum jon boat with 50 hp outboard tiller engine. The vessel was driven with the flow of the river in a backward orientation at approximately 2 km/h during sampling. The trawl was released slowly from the bow and allowed to settle on the bottom. Once bottom contact was made, the trawl was dragged for approximately 50 to 100 m along the bottom before retrieval. Two 50 x 30 cm otter doors (weighing 6 kg each) and a ¼” link chain attached to the 2.4 m footrope ensured bottom contact. A 2.4 m headrope, equipped with small plastic floats, opened the trawl mouth vertically. An inner cod end with 18 mm sapphire twine mesh collected larger fishes and debris (wood, rocks / boulders / shopping carts) and allowed smaller fishes to pass through to the outer cod end (3 mm mesh), thereby protecting small-bodied fishes from physical damage. The overall dimensions of the Gerken Siamese trawl were 2.4 m wide by 4.3 m deep (headrope to cod-end), and the length of the two tow ropes used to attach the trawl to the boat were adjusted based on depth of sampled area, where the tow rope length is seven times the water depth (Guy et al. 2009; Barnucz and Drake 2020). The total number of trawl tows per day was recorded. Other effort statistics, such as trawl distance or duration, were not recorded given the opportunistic sampling approach. Enumeration of Fishes Following capture by seine or trawl, fishes were immediately placed in aquaria with fresh river water to allow species sorting. Due to personnel and time constraints, it was not possible to identify and enumerate all fishes captured; instead, non-target species were released immediately and captured Eastern Sand Darter were held in a bucket of fresh river water and transferred to flow-through or aerated holding tanks prior to physiology experiments. On most sampling days, the capture of other darter species was documented in field notes. All captured Eastern Sand Darter were enumerated daily. When more individuals were captured than required for the day’s experiments, the surplus fish were released. Each Eastern Sand Darter used for an experimental trial was measured for total length (TL; mm), and mass (g) was recorded using a Ohaus® Valor ® 3000 Xtreme Compact Washdown Scale (300 g capacity); in

general, total length and mass measurements were not made on surplus fish except for fish captured in the July sampling period that were measured before release. All Eastern Sand Darter (surplus or otherwise) were released in approximately the same area where they were captured.

TAGGING Eastern Sand Darter selected for physiology experiments were tagged using Northwest Marine Technology Visible Implant Elastomer (VIE) tags on the ventral surface near the anal fin. Tagging occurred after each experimental trial was complete and once fish demonstrated signs of recovery (e.g., normalized swimming). The tag was used to mark individual fish to ensure the individual was not used for another physiology experiment within the same sampling period. Tag colour and location varied by sampling period (Table 1, Appendix 1) and was recorded for each individual fish. When tagged fish were recaptured, the tag colour and location of the recaptured fish were recorded. Tagged Eastern Sand Darter were held in aerated recovery aquaria for a minimum of 30 minutes prior to release to assess tagging mortality. Eastern Sand Darter captured but not used for the experiment were not tagged before release.

HABITAT SAMPLING Minimal habitat sampling was conducted for this study. Surface water temperature (°C), conductivity (µS), turbidity (NTU), and dissolved oxygen (mg/L) were measured approximately 0.2 m beneath the water’s surface using a YSI EX02 multiparameter sonde, which was deployed and allowed to stabilize for approximately 1 minute before measurements were recorded. These water quality parameters were taken once per sampling day after daily sampling was completed, near the centroid of the sampled area. Water velocity was not required for this study, however, it was noted that major rainfall events occurred during several sampling periods and affected flow and turbidity. Therefore, real-time hydrometric river discharge (m3/s) data was extracted from the Environment and Climate Change Canada (ECCC) Real Time Hydrometric Data web site (https://wateroffice.ec.gc.ca/mainmenu/real_time_data_index_e.html) on November 6, 2020 for the Grand River at Brantford (02GB001) station, approximately 5.5 km upstream of Cockshutt Bridge. Temperature loggers Two Onset HOBO Water Temperature Pro v2 data loggers were deployed for long-term in- stream measurement of water temperature at the upstream and downstream extent of accessible shoreline in June to monitor water temperature at 30 minute intervals throughout the study timeframe.

SAMPLING PERMITS AND DATA ARCHIVING Sampling for this project was conducted under SARA Permit Number 19-PCAA-00021. Seining, trawling, fish anesthesia, and tagging were conducted under Animal Use Protocol AUP 1968 and Standard Operating Protocols GWACC-116, GWACC-113, GWACC-105, and GWACC- 130, approved by the DFO and Environment and Climate Change Canada Animal Care Committee (operated under approval of the Canadian Council on Animal Care). Data associated with the collections in this report are housed under the project code “2019-ESDPT” in the Biodiversity Science database within the Great Lakes Laboratory for Fisheries and Aquatic Sciences. Every effort has been made to ensure the accuracy of data contained in this report; 3

however, results may be updated as part of ongoing data verification procedures. Data associated with this report may be obtained by contacting the Great Lakes Laboratory for Fisheries and Aquatic Sciences.

RESULTS

FISH ASSEMBLAGE SAMPLING In total, sampling occurred over 28 days across eight sampling periods in 2019 (Table 1, Appendix 2), resulting in the capture of 521 Eastern Sand Darter (Figure 3). Of these, 455 Eastern Sand Darter were captured by seining over 24 sampling days and 187 seine hauls and 66 Eastern Sand Darter were captured by trawling over 4 sampling days in 61 tows. Eastern Sand Darter was most frequently captured upstream of Cockshutt Bridge, downstream of the large riffle upstream of the bridge; however, the nearshore and thalweg areas downstream of the bridge also produced Eastern Sand Darter during some sampling periods (Figure 4). Seining was generally effective at capturing Eastern Sand Darter in wadeable habitats, though capture rates varied throughout sampling and decreased sharply late in the year. The number of Eastern Sand Darter per haul was not recorded; however, the fewest seine hauls to catch the minimum required number of Eastern Sand Darter was seven hauls, which captured 63 Eastern Sand Darter over three days in August. Eastern Sand Darter captures were most abundant in July, when 123 fish were captured in 12 seine hauls over four days. In contrast, far more seine hauls were conducted in late October but captured fewer Eastern Sand Darter. For example, on October 23rd field technicians conducted more than 60 seine hauls over a large area and did not capture the minimum target number of Eastern Sand Darter. On October 24th 18 seine hauls produced only a single Eastern Sand Darter, at which time trawling was initiated to access deeper water. Trawling effort was high to capture the minimum number of Eastern Sand Darter per day. Each of the four days in October and November in which trawling occurred required greater than ten trawl tows to capture at least ten Eastern Sand Darter. However, trawling allowed for effective fishing in deep water, ultimately provided operational advantages over seining during cooler periods. Sampling detected primarily adult Eastern Sand Darter (Table 2, Figure 5, Figure 6, Appendix 3). Across all sampling, mean total length was 62.2 mm (range: 46 – 72 mm TL) and mean weight was 1.4 g (range: 0.5 – 2.1 g; Figure 5). The mean total length of Eastern Sand Darter did not vary greatly between sampling visits (Figure 7); however, the largest individual (72 mm, 2.1 g) was captured in June and the smallest two individuals were captured in early October (46 mm, 0.6 g) and late October (48 mm, 0.5 g). The small individuals captured in the fall were likely age-0 individuals hatched earlier in 2019. Other darter species encountered during sampling were Greenside Darter (Etheostoma blennioides), Rainbow Darter (Etheostoma caeruleum), Johnny Darter (Etheostoma nigrum), Logperch (Percina caprodes), and Blackside Darter (Percina maculata) (Appendix 4).

TAGGING In total, 282 tags were implanted in Eastern Sand Darter (Appendix 3). Based on observation during the recovery period, there were no mortalities of tagged individuals, and all tagged individuals were successfully released. Thirty-seven recapture events occurred in which tagged Eastern Sand Darter were recaptured on at least one occasion after initially tagged. Thirty-two 4

were recaptured on one occasion (i.e. found once more after initial tagging; Figure 8a), 11 of which were recaptured within the same sampling period and did not receive another tag. Three individuals were recaptured on two occasions (Figure 8b) with one individual initially captured and tagged in September and recaptured on October 21 or 22 in the seine and again on October 28 in the trawl. Two more individuals were recaptured in two consecutive sampling periods after initial capture in July and August. Finally, one individual was recaptured on three occasions and therefore detected during four sampling periods (initial capture in June, recaptured in September, early October, and November). Trawling in late October and November resulted in the recapture of nine individuals that were previously captured by seining in shallower water, including the individual mentioned above.

HABITAT SAMPLING Mean water temperature by sampling period ranged from 6.25°C in November to 24.23°C in July (Table 3). The lowest daily water temperature in which Eastern Sand Darter were captured was 5.72°C on November 6and the highest water temperature in which Eastern Sand Darter was captured was 24.49°C on July 16 (Appendix 5). The remaining water quality parameters were inconsistently measured in different sampling periods as they were not critical for study objectives (Table 3, Appendix 5). Mean conductivity per sampling period ranged from a low of 478.5 µS in November to a high of 831.3 µS in August (daily range: 474.5 µS on November 5 – 838.0 µS on July 16). Mean dissolved oxygen per sampling period ranged from 6.8 mg/L in July to 12.4 mg/L in November (daily range: 6.07 mg/L on July 17 – 12.78 on November 6). Dissolved oxygen readings were not obtained for several sampling days in late October. The lowest mean turbidity by sampling period was 2.4 NTU for the late October sampling period; however turbidity was only measured in October on two of five days, prior to a rain event. Highest mean turbidity by sampling period was 12.5 NTU in September, with the highest daily turbidity measured on September 12 at 37.0 NTU. It is important to note that turbidity was not measured in late October after an extremely large rain event. Daily river discharge (m3/s) data from the ECCC hydrometric monitoring station along with the 2019 sampling periods is displayed Figure 9. The mean river discharge across all sampling events was 44.31 m3/s. River discharge was lowest in August sampling (mean 22.10 m3/s), and was highest in November sampling (73.55 m3/s; Table 3). The highest daily discharge experienced while sampling was 83.34 m3/s on November 4, and lowest was 21.85 m3/s on August 13 (Appendix 5). All sample periods except for August experienced a discharge event.

ACKNOWLEDGEMENTS We sincerely thank the individuals who were involved in field sampling: Jason Barnucz, Jacob Burbank, Megan Cowperthwaite, Britney Firth, Katelyn Gao, Megan Hutchings, Jared Skeath, Kurtis Smith, Sarah Young, and Charles-Andre Timmerman. Thank you to Julia Colm for developing Figures 1 and 2 in this report, and to Amy Boyko and Al Dextrase for their helpful reviews. Funding for this project was provided by DFO’s Species at Risk Program.

REFERENCES Barnucz, J. and Drake, D.A.R. 2020. Targeted surveys for Northern Madtom (Noturus stigmosus) in the lower East Sydenham River, Ontario, 2019. Can. Data Rep. Fish. Aquat. Sci. 1317: vi + 15 p. COSEWIC. 2009. COSEWIC assessment and status report on the Eastern Sand Darter Ammocrypta pellucida, Ontario populations and Quebec populations, in Canada. Committee on the Status of Endangered Wildlife in Canada. Ottawa. vii + 49 pp. (www.sararegistry.gc.ca/status/status_e.cfm). DFO. 2011. Recovery Potential Assessment of Eastern Sand Darter (Ammocrypta pellucida) in Canada. DFO Can. Sci. Advis. Sec. Sci. Advis. Rep. 2011/020. Dextrase, A.J., Mandrak, N.E., and Schaefer, J.A. 2014. Modelling occupancy of an imperiled stream fish at multiple scales while accounting for imperfect detection: implications for conservation. Freshw. Biol. 59:9 pp 1799-1815 Guy, C. S., Braaten, P.J., Herzog, D. P., Pitlo, J., and Rogers, R.S. 2009. Warmwater fish in rivers. Pages 59-84 in S. A. Bonar, W. A. Hubert, and D. W. Willis, editors. Standard methods for sampling North American freshwater fishes. American Fisheries Society, Bethesda, Maryland.

Figure 1. Detections of Eastern Sand Darter in the Grand River by DFO for the period 2004-2018 by seine net and benthic trawling. All sampling in 2019 between May and November was conducted at the 2019 study site at Cockshutt Bridge. 7

Figure 2. Detections of Eastern Sand Darter by DFO 2004-2018 at Cockshutt Bridge (Cockshutt Rd. and Erie Ave., Brantford), representing 15% of DFO’s total Eastern Sand Darter detections in the Grand River (n = 291 Eastern Sand Darter in 14 sampling events).

Figure 3. Photograph of Eastern Sand Darter (Ammocrypta pellucida) captured by seining in June 2019. Orange VIE tag on left of anal fin visible through translucent body.

Figure 4. Field sketches indicating area sampled by seine on a) July 16, 2019, b) October 2, 2019, and by trawl on c) October 28, 2019. The large riffle in the collection site is indicated by four wavy lines, shown perpendicular to river flow.

2.5

1.5

1 TotalWeight (g)

0.5

0 40 45 50 55 60 65 70 75 Total Length (mm) Figure 5. Relationship between total length (mm) and mass (g) of Eastern Sand Darter captured between May and November in the Grand River, Ontario, 2019 (n = 304).

45 40 35 30 25 20 15

10 NumberESD of Captured 5 0 40 45 50 55 60 65 70 75 Total Length (mm)

Figure 6. Length-frequency of Eastern Sand Darter (ESD) captured in the Grand River, Ontario between May and November, 2019 (n = 304). Overall, the mean total length (mm) of all Eastern Sand Darter captured was 62.69 (range: 46 – 72 mm).

10 10 10 10 8 8 8 8 6 6 6 6

4 4 4 4

NumberESD of NumberESD of NumberESD of NumberESD of 2 2 2 2 0 0 0 0 40 50 60 70 40 50 60 70 40 50 60 70 40 50 60 70 Total Length (mm) Total Length (mm) Total Length (mm) Total Length (mm)

a) May mean TL=65.3 mm (n=4) b) June mean TL=62.9 mm (n=48) c) July mean TL=59.3 mm (n=46) d) August mean TL=63.2 mm (n=45)

10 10 10 10 8 8 8 8 6 6 6 6

4 4 4 4

NumberESD of NumberESD of NumberESD of NumberESD of 2 2 2 2 0 0 0 0 40 50 60 70 40 50 60 70 40 50 60 70 40 50 60 70 Total Length (mm) Total Length (mm) Total Length (mm) Total Length (mm)

e) September mean TL=63.0 mm (n=45) f) Early October mean TL=63.0 mm (n=44) g) Late October mean TL=61.1 mm (n=40) h) November mean TL=62.7 mm (n=32)

Figure 7. Length-frequency of Eastern Sand Darter (ESD) captured in each sampling period between May and November, 2019 in the Grand River, Ontario. Mean total length (TL) provided below each panel.

Figure 8. Photographs of VIE tagged Eastern Sand Darter recaptured a) once in another sampling period (first capture in August, recaptured in September), and b) twice in two other sampling periods (first capture in July, recaptured in August and September).

Station (02GB001) data May June July August September early October late October November

180 160 140 120 100 80

60 Discharge (m3/s) 40 20 0

Date and time of hydrometric station sensor reading

Figure 9. Semi-hourly river discharge (m3/s) sensor data from ECCC Hydrometric Station (02GB001) on the Grand River at Brantford, approximately 5.5 km upstream of 2019 sampling site. Sampling periods are highlighted, indicating dates and times during which sampling for Eastern Sand Darter was conducted at Cockshutt Bridge.

Table 1. Summary of sampling periods, effort, gear type, and number of Eastern Sand Darter captured. Effort is displayed as number of hauls or tows for each gear type, seine net (SN) and benthic trawl (TRL), respectively.

Sampling Sampling Gear Total effort Eastern Sand period Dates days type (hauls/tows) Darter captured May May 22, 2019 1 SN 8 4 June June 3-6, 2019 4 SN 18 64 July July 15-18, 2019 4 SN 12 123 August August 12-14, 2019 3 SN 7 63 September September 9-12, 2019 4 SN 15 84 Early October September 30-October 3, 2019 4 SN 20 82 October 21-24, 2019 4 SN 107 35 Late October October 28, 2019 1 TRL 15 24 November November 4-6, 2019 3 TRL 46 42 24 SN 187 455 Overall by gear type 4 TRL 61 66 Total Eastern Sand Darter captured 521

Table 2. Summary of total length (mm) and mass (g) of Eastern Sand Darter captured by sampling period.

Number of Total length (mm) Mass (g) Sampling Eastern period Sand Darter Min. Max. Mean Min. Max. Mean May 4 63 68 65.3 1.2 1.5 1.35 June 48 51 72 62.9 0.6 2.1 1.39 July 46 50 70 59.3 0.7 2.0 1.22 August 45 55 69 63.2 1.0 2.0 1.44 September 45 59 69 63.0 1.2 1.9 1.50 Early October 44 48 70 63.0 0.5 1.9 1.41 Late October 40 46 67 61.1 0.6 1.8 1.34 November 32 59 69 62.7 1.2 1.9 1.47 Overall 304 46 72 62.2 0.5 2.1 1.39

Table 3. Summary of mean abiotic habitat values over each sampling period. Mean river discharge calculated from semi-hourly discharge sensor data extracted from Environment and Climate Change Canada real-time hydrometric monitoring station website.

Air Water Dissolved Mean river Sampling temperature temperature Conductivity Oxygen Turbidity discharge period (°C) (°C) (µS) (mg/L) (NTU) (m3/s) May - 14.89 - 10.5 - 51.64 June 17.1 16.22 611.0 9.1 6.0 53.09 July 25.1 24.23 795.5 6.8 8.9 34.09 August 22.5 23.17 831.3 7.0 7.3 22.10 September 16.1 18.81 730.8 8.5 12.5 31.29 Early October 17.1 17.16 711.5 8.4 9.1 35.39 Late October 12.7 11.59 632.5 11.3 2.4 38.68 November 6.0 6.07 478.5 12.4 11.0 73.55 Overall 18.2 16.84 713.9 8.7 8.8 44.31

Appendix 1. Visible Implant Elastomer (VIE) tag colour and location codes for Eastern Sand Darter assessed in physiology experiments by sampling period.

Sampling period Tag colour Locaton of tag May Not tagged N/A June Orange Left of anal fin July Blue Right of anal fin July Blue Left of anal fin August Orange Right of anal fin September Yellow Left of anal fin Early October Yellow Right of anal fin Late October Red Right of anal fin November Red Left of anal fin

Appendix 2. Sampling date, field code, sampling duration, gear type, and fishing effort for captures of Eastern Sand Darter (ESD) in the Grand River, 2019, to support physiology experiments. Where gear code SN = Seine (bag seine, 9.14 x1.8 m, 3 mm mesh), and TRL = Trawl (Gerken Siamese Trawl, 2.4 m foot rope, 3 mm outer cod mesh).

Sampling Sampling Effort Total Sampling start time stop time Gear (hauls or ESD date Unique field code (24h) (24h) code Description of gear tows) captured 22-May-19 2019-ESDPT-220519-001A 10:00 12:00 SN Bag seine 8 4 03-Jun-19 2019-ESDPT-030619-001A 08:30 09:25 SN Bag seine 4 16 04-Jun-19 2019-ESDPT-040619-001A 07:50 08:16 SN Bag seine 2 21 05-Jun-19 2019-ESDPT-060619-001A 07:40 08:50 SN Bag seine 8 18 06-Jun-19 2019-ESDPT-060619-002A 07:45 08:00 SN Bag seine 2 9 15-Jul-19 2019-ESDPT-150719-001A 09:08 09:50 SN Bag seine 4 19 16-Jul-19 2019-ESDPT-160719-001A 07:50 08:35 SN Bag seine 5 58 17-Jul-19 2019-ESDPT-170719-001A 08:15 08:40 SN Bag seine 2 33 18-Jul-19 2019-ESDPT-180719-001A 07:50 08:08 SN Bag seine 1 13 12-Aug-19 2019-ESDPT-120819-001A 08:30 08:40 SN Bag seine 1 26 13-Aug-19 2019-ESDPT-130819-001A 07:40 08:00 SN Bag seine 3 20 14-Aug-19 2019-ESDPT-140819-001A 07:36 07:55 SN Bag seine 3 17 09-Sep-19 2019-ESDPT-090919-001A 09:02 09:35 SN Bag seine 3 36 10-Sep-19 2019-ESDPT-100919-001A 08:04 08:29 SN Bag seine 3 19 11-Sep-19 2019-ESDPT-110919-001A 07:51 08:40 SN Bag seine 5 17 12-Sep-19 2019-ESDPT-120919-001A 08:10 08:50 SN Bag seine 4 12 30-Sep-19 2019-ESDPT-300919-001A 08:41 08:54 SN Bag seine 1 36 01-Oct-19 2019-ESDPT-011019-001A 07:49 08:43 SN Bag seine 5 18 02-Oct-19 2019-ESDPT-021019-001A 08:00 09:50 SN Bag seine 12 23 03-Oct-19 2019-ESDPT-031019-001A 07:45 07:57 SN Bag seine 2 5 21-Oct-19 2019-ESDPT-211019-001A 08:41 09:45 SN Bag seine 7 10 22-Oct-19 2019-ESDPT-221019-001A 07:40 10:40 SN Bag seine 22 15 23-Oct-19 2019-ESDPT-231019-001A 07:58 16:00 SN Bag seine >60 9 24-Oct-19 2019-ESDPT-241019-001A 09:01 12:30 SN Bag seine 18 1 28-Oct-19 2019-ESDPT-281019-001A 08:45 12:00 TRL Gerken Siamese trawl 14 24 04-Nov-19 2019-ESDPT-041119-001A 08:40 11:33 TRL Gerken Siamese trawl 22 10 05-Nov-19 2019-ESDPT-051119-001A 07:15 08:45 TRL Gerken Siamese trawl 10 13 06-Nov-19 2019-ESDPT-061119-001A 07:45 10:15 TRL Gerken Siamese trawl 14 19

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-05-22 220519-001 68 1.3 Y N N N/A N N/A 2019-05-22 220519-002 66 1.5 Y N N N/A N N/A 2019-05-22 220519-003 63 1.2 Y N N N/A N N/A 2019-05-22 220519-004 64 1.4 Y N N N/A N N/A 2019-06-03 030619-001 67 1.7 Y N Y Orange - Left N N/A 2019-06-03 030619-002 62 1.3 Y N Y Orange - Left N N/A 2019-06-03 030619-003 61 1.1 Y N Y Orange - Left N N/A 2019-06-03 030619-004 64 1.6 Y N Y Orange - Left N N/A 2019-06-03 030619-005 66 1.7 Y N Y Orange - Left N N/A 2019-06-03 030619-006 59 0.7 Y N Y Orange - Left N N/A 2019-06-03 030619-007 68 1.9 N Y N N/A N N/A 2019-06-03 030619-008 62 1.4 Y N Y Orange - Left N N/A 2019-06-03 030619-009 64 1.4 Y N Y Orange - Left N N/A 2019-06-03 030619-010 59 0.9 Y N Y Orange - Left N N/A 2019-06-04 040619-001 65 1.5 Y N Y Orange - Left N N/A 2019-06-04 040619-002 63 1.2 Y N Y Orange - Left N N/A 2019-06-04 040619-003 61 1.4 Y N Y Orange - Left N N/A 2019-06-04 040619-004 61 1.3 Y N Y Orange - Left N N/A 2019-06-04 040619-005 63 1.6 Y N Y Orange - Left N N/A 2019-06-04 040619-006 67 1.5 Y N Y Orange - Left N N/A 2019-06-04 040619-007 65 1.4 Y N Y Orange - Left N N/A 2019-06-04 040619-008 62 1.5 Y N Y Orange - Left N N/A 2019-06-04 040619-009 65 1.2 Y N Y Orange - Left N N/A 2019-06-04 040619-010 67 1.6 Y N Y Orange - Left N N/A 2019-06-04 040619-011 68 1.6 Y N Y Orange - Left N N/A 2019-06-04 040619-012 65 1.5 Y N Y Orange - Left N N/A 2019-06-04 040619-013 60 1.2 Y N Y Orange - Left N N/A 2019-06-04 040619-014 55 0.8 Y N Y Orange - Left N N/A 2019-06-05 050619-001 65 1.0 Y N Y Orange - Left N N/A 2019-06-05 050619-002 63 1.4 Y N Y Orange - Left N N/A 2019-06-05 050619-003 66 1.8 Y N Y Orange - Left N N/A 2019-06-05 050619-004 62 1.3 Y N Y Orange - Left N N/A 2019-06-05 050619-005 51 0.6 Y N Y Orange - Left N N/A 2019-06-05 050619-006 72 2.1 Y N Y Orange - Left N N/A 2019-06-05 050619-007 67 1.6 Y N Y Orange - Left N N/A 2019-06-05 050619-008 63 1.3 Y N Y Orange - Left N N/A 2019-06-05 050619-009 61 1.1 Y N Y Orange - Left N N/A 2019-06-05 050619-010 60 1.2 Y N Y Orange - Left N N/A 2019-06-05 050619-011 63 1.4 Y N Y Orange - Left N N/A 2019-06-05 050619-012 68 1.6 Y N Y Orange - Left N N/A 2019-06-05 050619-013 63 1.3 Y N Y Orange - Left N N/A 2019-06-05 050619-014 65 1.7 Y N Y Orange - Left N N/A 2019-06-05 050619-015 65 1.6 Y N Y Orange - Left N N/A 2019-06-06 060619-001 64 1.5 Y N Y Orange - Left N N/A 2019-06-06 060619-002 65 1.7 Y N Y Orange - Left N N/A 2019-06-06 060619-003 66 1.6 Y N Y Orange - Left N N/A

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-06-06 060619-004 53 0.8 Y N Y Orange - Left N N/A 2019-06-06 060619-005 65 1.7 Y N Y Orange - Left N N/A 2019-06-06 060619-006 64 1.6 Y N Y Orange - Left N N/A 2019-06-06 060619-007 64 1.6 Y N Y Orange - Left N N/A 2019-06-06 060619-008 64 1.7 Y N Y Orange - Left N N/A 2019-06-06 060619-009 51 0.6 Y N Y Orange - Left N N/A 2019-07-15 150719-001 56 0.9 N Y N N/A N N/A 2019-07-15 150719-002 55 0.9 Y N Y Blue - Right N N/A 2019-07-15 150719-003 54 0.9 Y N Y Blue - Right N N/A 2019-07-15 150719-004 50 0.7 Y N Y Blue - Right N N/A 2019-07-15 150719-005 62 1.5 Y N Y Blue - Right N N/A 2019-07-15 150719-006 60 1.2 Y N Y Blue - Right N N/A 2019-07-15 150719-007 60 1.3 Y N Y Blue - Right N N/A 2019-07-15 150719-008 64 1.2 N Y N N/A N N/A 2019-07-15 150719-009 56 0.9 Y N Y Blue - Right N N/A 2019-07-15 150719-010 65 1.3 Y N Y Blue - Right N N/A 2019-07-15 150719-011 59 1.2 Y N Y Blue - Right N N/A 2019-07-15 150719-012 55 0.9 Y N Y Blue - Left N N/A 2019-07-15 150719-013 62 1.4 Y N Y Blue - Left Y Orange - Left 2019-07-15 150719-014 61 1.2 Y N Y Blue - Left N N/A 2019-07-15 150719-015 61 1.1 Y N Y Blue - Left N N/A 2019-07-16 160719-001 53 0.9 Y N Y Blue - Right N N/A 2019-07-16 160719-002 55 0.9 Y N Y Blue - Right N N/A 2019-07-16 160719-003 64 1.5 Y N Y Blue - Right N N/A 2019-07-16 160719-004 57 1.0 Y N Y Blue - Right N N/A 2019-07-16 160719-005 61 1.3 N Y N N/A N N/A 2019-07-16 160719-006 60 1.1 N Y N N/A N N/A 2019-07-16 160719-007 56 1.0 Y N Y Blue - Right N N/A 2019-07-16 160719-008 54 0.9 N Y N N/A N N/A 2019-07-16 160719-009 55 1.0 Y N Y Blue - Right N N/A 2019-07-16 160719-010 61 1.3 Y N Y Blue - Right N N/A 2019-07-16 160719-011 62 1.4 Y N Y Blue - Right N N/A 2019-07-16 160719-012 51 0.8 N Y N N/A N N/A 2019-07-16 160719-013 54 1.0 Y N Y Blue - Right N N/A 2019-07-16 160719-014 62 1.3 Y N Y Blue - Right N N/A 2019-07-16 160719-015 56 1.0 Y N Y Blue - Right N N/A 2019-07-16 160719-016 54 0.9 Y N Y Blue - Right N N/A 2019-07-16 160719-017 62 1.3 N Y N N/A N N/A 2019-07-17 170719-001 56 1.0 Y N Y Blue - Right N N/A 2019-07-17 170719-002 65 1.8 Y N Y Blue - Right N N/A 2019-07-17 170719-003 53 0.9 Y N Y Blue - Right N N/A 2019-07-17 170719-004 60 1.3 Y N Y Blue - Right N N/A 2019-07-17 170719-005 56 1.1 Y N Y Blue - Right N N/A 2019-07-17 170719-006 64 1.6 Y N Y Blue - Right N N/A 2019-07-17 170719-007 57 1.2 Y N Y Blue - Right N N/A 2019-07-17 170719-008 63 1.3 Y N Y Blue - Right N N/A 2019-07-17 170719-009 63 1.6 Y N Y Blue - Right N N/A 2019-07-17 170719-010 63 1.4 Y N Y Blue - Right N N/A 2019-07-17 170719-011 59 1.2 Y N Y Blue - Right N N/A 2019-07-18 180719-001 62 1.5 Y N Y Blue - Right N N/A 2019-07-18 180719-002 62 1.3 Y N Y Blue - Right N N/A 2019-07-18 180719-003 69 1.9 Y N Y Blue - Right N N/A 2019-07-18 180719-004 70 2.0 Y N Y Blue - Right N N/A 2019-07-18 180719-005 62 1.5 Y N Y Blue - Right N N/A 2019-07-18 180719-006 66 1.8 Y N Y Blue - Right N N/A 2019-07-18 180719-007 59 1.0 Y N Y Blue - Right N N/A

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-08-12 120819-001 64 1.3 Y N Y Orange - Right N N/A 2019-08-12 120819-002 63 1.5 Y N Y Orange - Right N N/A 2019-08-12 120819-003 65 1.6 Y N Y Orange - Right N N/A 2019-08-12 120819-004 64 1.5 Y N Y Orange - Right Y Blue - Left 2019-08-12 120819-005 66 1.6 Y N Y Orange - Right N N/A 2019-08-12 120819-006 65 1.5 Y N Y Orange - Right N N/A 2019-08-12 120819-007 64 1.2 Y N Y Orange - Right N N/A 2019-08-12 120819-008 67 1.7 Y N Y Orange - Right N N/A 2019-08-12 120819-009 65 1.7 Y N Y Orange - Right Y Blue - Left 2019-08-12 120819-010 66 1.5 Y N Y Orange - Right N N/A 2019-08-12 120819-011 61 1.3 Y N Y Orange - Right N N/A 2019-08-12 120819-012 60 1.3 Y N Y Orange - Right N N/A 2019-08-13 130819-001 55 1.2 Y N Y Orange - Right N N/A 2019-08-13 130819-002 62 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-003 61 1.3 Y N Y Orange - Right N N/A 2019-08-13 130819-004 59 1.2 Y N Y Orange - Right N N/A 2019-08-13 130819-005 68 1.7 Y N Y Orange - Right N N/A 2019-08-13 130819-006 65 1.6 Y N Y Orange - Right N N/A 2019-08-13 130819-007 61 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-008 60 1.3 Y N Y Orange - Right N N/A 2019-08-13 130819-009 63 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-010 65 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-011 66 1.8 Y N Y Orange - Right N N/A 2019-08-13 130819-012 62 1.3 N Y N N/A N N/A 2019-08-13 130819-013 64 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-014 64 1.5 Y N Y Orange - Right N N/A 2019-08-13 130819-015 58 1.2 Y N Y Orange - Right N N/A 2019-08-13 130819-016 58 1.1 Y N Y Orange - Right N N/A 2019-08-13 130819-017 61 1.3 Y N Y Orange - Right N N/A 2019-08-14 140819-001 62 1.4 N Y N N/A N N/A 2019-08-14 140819-002 67 1.7 Y N Y Orange - Right N N/A 2019-08-14 140819-003 66 1.6 Y N Y Orange - Right Y Blue - Right 2019-08-14 140819-004 65 1.6 Y N Y Orange - Right N N/A 2019-08-14 140819-005 63 1.4 N Y N N/A N N/A 2019-08-14 140819-006 65 1.5 Y N Y Orange - Right N N/A 2019-08-14 140819-007 69 2.0 Y N Y Orange - Right N N/A 2019-08-14 140819-008 30 1.2 Y N Y Orange - Right N N/A 2019-08-14 140819-009 66 1.5 Y N Y Orange - Right N N/A 2019-08-14 140819-010 63 1.3 Y N Y Orange - Right N N/A 2019-08-14 140819-011 61 1.1 Y N Y Orange - Right N N/A 2019-08-14 140819-012 67 1.7 Y N Y Orange - Right N N/A 2019-08-14 140819-013 67 1.8 Y N Y Orange - Right N N/A 2019-08-14 140819-014 61 1.1 Y N Y Orange - Right N N/A 2019-08-14 140819-015 60 1.1 Y N Y Orange - Right N N/A 2019-08-14 140819-016 58 1.0 Y N Y Orange - Right N N/A 2019-09-09 090919-001 68 1.9 Y N Y Yellow - Left N N/A 2019-09-09 090919-002 60 1.5 Y N Y Yellow - Left N N/A 2019-09-09 090919-003 61 1.4 Y N Y Yellow - Left N N/A 2019-09-09 090919-004 62 1.4 Y N Y Yellow - Left N N/A 2019-09-09 090919-005 63 1.5 Y N Y Yellow - Left N N/A 2019-09-09 090919-006 62 1.6 Y N Y Yellow - Left N N/A 2019-09-09 090919-007 62 1.5 Y N Y Yellow - Left N N/A 2019-09-09 090919-008 65 1.6 Y N Y Yellow - Left Y Orange - Left 2019-09-09 090919-009 60 1.2 Y N Y Yellow - Left N N/A 2019-09-10 100919-001 61 1.4 Y N Y Yellow - Left N N/A 2019-09-10 100919-002 63 1.5 Y N Y Yellow - Left N N/A

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-09-10 100919-003 61 1.4 Y N Y Yellow - Left N N/A 2019-09-10 100919-004 63 1.5 Y N Y Yellow - Left N N/A 2019-09-10 100919-005 64 1.6 Y N Y Yellow - Left Y Blue - Left, Orange - Right 2019-09-10 100919-006 62 1.4 Y N Y Yellow - Left N N/A 2019-09-10 100919-007 61 1.3 Y N Y Yellow - Left N N/A 2019-09-10 100919-008 64 1.6 Y N Y Yellow - Left N N/A 2019-09-10 100919-009 61 1.4 Y N Y N/A Y* Yellow - Left 2019-09-10 100919-010 64 1.7 Y N Y Yellow - Left N N/A 2019-09-10 100919-011 66 1.6 Y N Y Yellow - Left N N/A 2019-09-10 100919-012 59 1.2 Y N Y Yellow - Left N N/A 2019-09-10 100919-013 60 1.3 Y N Y Yellow - Left Y Orange - Right 2019-09-10 100919-014 65 1.6 Y N Y Yellow - Left N N/A 2019-09-10 100919-015 63 1.5 Y N Y Yellow - Left N N/A 2019-09-10 100919-016 - - Y N N N/A Y* Yellow - Left 2019-09-10 100919-017 - - Y N N N/A Y* Yellow - Left 2019-09-11 110919-001 65 1.6 Y N Y Yellow - Left N N/A 2019-09-11 110919-002 65 1.6 Y N Y Yellow - Left N N/A 2019-09-11 110919-003 61 1.4 Y N Y Yellow - Left N N/A 2019-09-11 110919-004 67 1.8 Y N Y Yellow - Left N N/A 2019-09-11 110919-005 64 1.5 Y N Y Yellow - Left N N/A 2019-09-11 110919-006 63 1.5 Y N Y Yellow - Left N N/A 2019-09-11 110919-007 64 1.5 Y N Y Yellow - Left N N/A 2019-09-11 110919-008 67 1.9 Y N Y Yellow - Left N N/A 2019-09-11 110919-009 61 1.4 Y N Y Yellow - Left N N/A 2019-09-11 110919-010 64 1.6 Y N Y Yellow - Left N N/A 2019-09-11 110919-011 63 1.5 Y N Y Yellow - Left N N/A 2019-09-11 110919-012 64 1.5 Y N Y Yellow - Left N N/A 2019-09-11 110919-013 65 1.6 Y N Y Yellow - Left N N/A 2019-09-11 110919-014 - - Y N N N/A Y* Yellow - Left 2019-09-11 110919-015 - - Y N N N/A Y* Yellow - Left 2019-09-11 110919-016 - - Y N N N/A Y* Yellow - Left 2019-09-12 120919-001 67 1.9 Y N Y Yellow - Left Y Orange - Right 2019-09-12 120919-002 59 1.4 Y N Y Yellow - Left N N/A 2019-09-12 120919-003 60 1.3 Y N Y Yellow - Left N N/A 2019-09-12 120919-004 61 1.3 Y N Y Yellow - Left N N/A 2019-09-12 120919-005 59 1.2 Y N Y Yellow - Left N N/A 2019-09-12 120919-006 62 1.5 Y N Y Yellow - Left N N/A 2019-09-12 120919-007 59 1.2 Y N Y Yellow - Left N N/A 2019-09-12 120919-008 65 1.7 Y N Y Yellow - Left N N/A 2019-09-30 300919-001 66 1.8 Y N Y Yellow - Right Y Yellow - Left 2019-09-30 300919-002 56 1.0 Y N Y Yellow - Right N N/A 2019-09-30 300919-003 63 1.5 Y N Y Yellow - Right N N/A 2019-09-30 300919-004 64 1.5 Y N Y Yellow - Right N N/A 2019-09-30 300919-005 65 1.6 N Y N N/A N N/A 2019-09-30 300919-006 61 1.6 Y N Y Yellow - Right N N/A 2019-09-30 300919-007 58 1.1 Y N Y Yellow - Right N N/A 2019-09-30 300919-008 48 0.5 Y N Y Yellow - Right N N/A 2019-09-30 300919-009 63 1.5 Y N Y Yellow - Right N N/A 2019-09-30 300919-010 61 1.3 N Y N N/A N N/A 2019-09-30 300919-011 - - Y N N N/A Y Orange - Right, Yellow - Left 2019-10-01 011019-001 69 1.8 Y N Y Yellow - Right N N/A 2019-10-01 011019-002 61 1.3 Y N Y Yellow - Right N N/A 2019-10-01 011019-003 59 1.2 Y N Y Yellow - Right N N/A 2019-10-01 011019-004 69 1.8 Y N Y Yellow - Right N N/A 2019-10-01 011019-005 67 1.9 Y N Y Yellow - Right N N/A 2019-10-01 011019-006 69 1.9 Y N Y Yellow - Right N N/A

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-10-01 011019-007 63 1.5 Y N Y Yellow - Right N N/A 2019-10-01 011019-008 64 1.8 Y N Y Yellow - Right N N/A 2019-10-01 011019-009 61 1.3 Y N Y Yellow - Right N N/A 2019-10-01 011019-010 69 1.7 Y N Y Yellow - Right N N/A 2019-10-01 011019-011 64 1.5 Y N Y Yellow - Right N N/A 2019-10-01 011019-012 63 1.3 Y N Y Yellow - Right N N/A 2019-10-01 011019-013 59 1.1 Y N Y Yellow - Right N N/A 2019-10-01 011019-014 61 1.1 Y N Y Yellow - Right N N/A 2019-10-01 011019-015 64 1.3 N Y N N/A N N/A 2019-10-02 021019-001 66 1.6 Y N Y Yellow - Right Y Orange - Right 2019-10-02 021019-002 63 1.3 Y N Y Yellow - Right N N/A 2019-10-02 021019-003 65 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-004 64 1.3 Y N Y Yellow - Right N N/A 2019-10-02 021019-005 64 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-006 65 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-007 60 1.2 Y N Y Yellow - Right N N/A 2019-10-02 021019-008 70 1.8 Y N Y Yellow - Right Y Orange - Right, Yellow - Left 2019-10-02 021019-009 66 1.6 Y N Y Yellow - Right Y Blue - Right 2019-10-02 021019-010 64 1.2 Y N Y Yellow - Right N N/A 2019-10-02 021019-011 64 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-012 65 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-013 63 1.5 Y N Y Yellow - Right N N/A 2019-10-02 021019-014 66 1.3 Y N Y Yellow - Right N N/A 2019-10-02 021019-015 60 1.1 Y N Y Yellow - Right N N/A 2019-10-02 021019-016 - - Y N N N/A Y* Yellow - Right 2019-10-02 021019-017 - - Y N N N/A Y* Yellow - Right 2019-10-02 021019-018 - - Y N N N/A Y* Yellow - Right 2019-10-03 031019-001 61 1.3 Y N Y Yellow - Right N N/A 2019-10-03 031019-002 60 1.3 Y N Y Yellow - Right N N/A 2019-10-03 031019-003 65 1.5 Y N Y Yellow - Right N N/A 2019-10-03 031019-004 62 1.3 Y N Y Yellow - Right N N/A 2019-10-03 031019-005 61 1.2 Y N Y Yellow - Right N N/A 2019-10-21 210119-001 62 0.8 Y N Y Red - Right N N/A 2019-10-21 210119-002 62 1.4 Y N Y Red - Right N N/A 2019-10-21 210119-003 61 1.3 Y N Y Red - Right N N/A 2019-10-21 210119-004 59 1.2 Y N Y Red - Right N N/A 2019-10-21 210119-005 49 0.6 Y N Y Red - Right N N/A 2019-10-21 210119-006 63 1.5 Y N Y Red - Right Y Yellow - Left 2019-10-21 210119-007 61 1.4 Y N Y Red - Right N N/A 2019-10-21 210119-008 61 1.4 Y N Y Red - Right N N/A 2019-10-21 210119-009 60 1.2 Y N Y Red - Right Y Yellow - Right 2019-10-21 210119-010 64 1.6 Y N Y Red - Right N N/A 2019-10-22 221019-001 63 1.5 Y N Y Red - Right Y Yellow - Left 2019-10-22 221019-002 65 1.5 Y N Y Red - Right N N/A 2019-10-22 221019-003 62 1.3 Y N Y Red - Right N N/A 2019-10-22 221019-004 62 1.2 Y N Y Red - Right N N/A 2019-10-22 221019-005 60 1.2 Y N Y Red - Right N N/A 2019-10-22 221019-006 63 1.5 Y N Y Red - Right N N/A 2019-10-22 221019-007 60 1.2 Y N Y Red - Right N N/A 2019-10-22 221019-008 61 1.4 Y N Y Red - Right N N/A 2019-10-22 221019-009 64 1.6 Y N Y Red - Right N N/A 2019-10-22 221019-010 61 1.3 Y N Y Red - Right N N/A 2019-10-22 221019-011 59 1.3 Y N Y Red - Right N N/A 2019-10-22 221019-012 62 1.5 Y N Y Red - Right N N/A 2019-10-22 221019-013 - - Y N N N/A Y* Red - Right 2019-10-23 231019-001 46 0.6 Y N Y Red - Right N N/A

Unique

individual Date code New tag code Existing tag code(s) captured (ESD-ID) TL M (colour-position) (colour - position)

RELEASED KEPT TAG NEW RECAP 2019-10-23 231019-002 63 1.4 Y N Y Red - Right N N/A 2019-10-23 231019-003 57 1.1 Y N Y Red - Right N N/A 2019-10-23 231019-004 60 1.2 Y N Y Red - Right N N/A 2019-10-23 231019-005 63 1.6 Y N Y Red - Right N N/A 2019-10-23 231019-006 59 1.3 Y N Y Red - Right N N/A 2019-10-23 231019-007 63 1.4 Y N Y Red - Right N N/A 2019-10-23 231019-008 62 1.3 Y N Y Red - Right N N/A 2019-10-23 231019-009 61 1.3 Y N N Red - Right Y N/A 2019-10-24 241019-001 - - Y N N N/A Y* Red - Right 2019-10-28 281019-001 61 1.4 Y N Y Red - Right N N/A 2019-10-28 281019-002 67 1.7 Y N Y Red - Right N N/A 2019-10-28 281019-003 65 1.0 Y N Y Red - Right N N/A 2019-10-28 281019-004 63 1.4 Y N Y Red - Right N N/A 2019-10-28 281019-005 66 1.6 Y N Y Red - Right N N/A 2019-10-28 281019-006 66 1.6 Y N Y Red - Right N N/A 2019-10-28 281019-007 67 1.8 Y N Y Red - Right N N/A 2019-10-28 281019-008 61 1.2 Y N Y Red - Right N N/A 2019-10-28 281019-009 61 1.2 Y N Y Red - Right N N/A 2019-10-28 281019-010 59 1.2 Y N Y Red - Right N N/A 2019-10-28 281019-011 - - N Y N N/A N N/A 2019-10-28 281019-012 - - Y N N N/A Y* Red - Right, Yellow - Left 2019-11-04 041119-001 62 1.4 Y N Y Red - Left, Red - N N/A Right 2019-11-04 041119-002 69 1.8 Y N Y Red - Left N N/A 2019-11-04 041119-003 64 1.7 Y N Y Red - Left N N/A 2019-11-04 041119-004 60 1.3 Y N Y Red - Left N N/A 2019-11-04 041119-005 66 1.8 Y N Y Red - Left N N/A 2019-11-04 041119-006 59 1.3 Y N Y Red - Left N N/A 2019-11-04 041119-007 61 1.4 Y N Y Red - Left N N/A 2019-11-04 041119-008 60 1.4 Y N Y Red - Left Y Yellow - Right 2019-11-04 041119-009 61 1.4 Y N Y Red - Left Y Red - Right 2019-11-04 041119-010 60 1.2 Y N Y Red - Left N N/A 2019-11-05 051119-001 66 1.6 Y N Y Red - Left Y Red - Right 2019-11-05 051119-002 65 1.4 Y N Y Red - Left Y Yellow - Left 2019-11-05 051119-003 61 1.4 Y N Y Red - Left N N/A 2019-11-05 051119-004 66 1.7 Y N Y Red - Left Y Yellow - Left 2019-11-05 051119-005 62 1.2 Y N Y Red - Left N N/A 2019-11-05 051119-006 59 1.2 Y N Y Red - Left N N/A 2019-11-05 051119-007 64 1.6 Y N Y Red - Left N N/A 2019-11-05 051119-008 63 1.4 Y N Y Red - Left N N/A 2019-11-05 051119-009 61 1.4 Y N Y Red - Left N N/A 2019-11-05 051119-010 60 1.3 Y N Y Red - Left N N/A 2019-11-05 051119-011 64 1.6 Y N Y Red - Left Y Orange - Right 2019-11-05 051119-012 62 1.4 Y N Y Red - Left N N/A 2019-11-05 051119-013 67 1.8 Y N Y Red - Left Y Yellow - Right 2019-11-06 061119-001 60 1.4 Y N N N/A N N/A 2019-11-06 061119-002 64 1.6 Y N N N/A N N/A 2019-11-06 061119-003 60 1.4 Y N N N/A N N/A 2019-11-06 061119-004 62 1.3 Y N N N/A N N/A 2019-11-06 061119-005 62 1.3 Y N N N/A N N/A 2019-11-06 061119-006 66 1.7 Y N N N/A N N/A 2019-11-06 061119-007 69 1.9 Y N N N/A Y Orange - Left, Yellow - Left, Yellow - Right 2019-11-06 061119-008 62 1.4 Y N N N/A N N/A 2019-11-06 061119-009 59 1.2 Y N N N/A N N/A 2019-11-06 061119-010 - - N Y N N/A N N/A

Appendix 4. Summary of field notes indicating detections of other darter species captured during targeted sampling for Eastern Sand Darter in the Grand River, Ontario, 2019, where 1 = detected, 0 = not detected. Shown are dates where detection of other darter species was documented.

maculat Sampling date

Etheostoma blennioides Etheostoma caeruleum Etheostoma nigrum Percina caprodes Percina 16-Jul-19 1 1 1 0 0 17-Jul-19 0 0 1 1 1 18-Jul-19 0 1 1 1 1 12-Aug-19 1 0 1 0 1 13-Aug-19 0 1 1 0 0 14-Aug-19 1 0 1 1 1 9-Sep-19 1 1 1 1 1 10-Sep-19 1 1 1 0 0 11-Sep-19 1 1 1 1 1 30-Sep-19 1 1 1 1 1 1-Oct-19 1 1 1 0 1 2-Oct-19 1 1 1 1 1 21-Oct-19 1 1 1 0 1 23-Oct-19 1 1 1 1 0

Appendix 5. Daily abiotic habitat assessment measurements, collected immediately after sampling using a YSI EXO2 multiparameter sonde. Mean river discharge during sampling data calculated by averaging semi-hourly sensor data extracted from Environment and Climate Change Canada Real-time Hydrometric Data website on November 6, 2020.

Mean Air Water Dissolved river temperature temperature Conductivity Oxygen Turbidity discharge Date (°C) (°C) (µS) (mg/L) (NTU) (m3/s) 22-May-19 - 14.89 - 10.5 - 51.64 03-Jun-19 13.9 15.33 - 9.72 - 58.00 04-Jun-19 14.5 15.55 611 9.10 6.00 49.50 05-Jun-19 22.9 17.20 - 8.90 - 46.33 06-Jun-19 17.0 16.78 - 8.60 - 61.90 15-Jul-19 25.1 23.65 816 7.55 5.45 26.25 16-Jul-19 28.7 24.49 838 6.83 5.54 25.90 17-Jul-19 23.3 24.39 819 6.07 5.14 31.20 18-Jul-19 23.1 24.40 709 6.56 19.50 57.10 12-Aug-19 25.3 23.02 830 8.02 7.33 22.00 13-Aug-19 22.3 23.15 830 6.57 6.00 21.85 14-Aug-19 20.0 23.35 834 6.41 8.70 22.70 09-Sep-19 - 17.76 752 9.37 2.90 21.80 10-Sep-19 - 17.38 736 8.84 5.50 22.00 11-Sep-19 - 19.51 752 8.10 4.70 23.77 12-Sep-19 16.1 20.60 683 7.56 37.00 56.70 30-Sep-19 16.2 15.59 731 8.90 4.10 22.30 01-Oct-19 20.2 17.45 764 8.38 7.00 22.33 02-Oct-19 19.0 18.91 723 7.92 10.37 44.40 03-Oct-19 13.0 16.69 628 8.52 15.00 51.60 21-Oct-19 14.2 12.93 632 12.65 2.10 27.10 22-Oct-19 14.3 12.82 633 9.93 2.70 31.23 23-Oct-19 - 11.09 - - - 31.96 24-Oct-19 - - - - - 29.69 28-Oct-19 9.7 9.50 - - - 69.04 04-Nov-19 - 5.71 - - - 83.34 05-Nov-19 6.0 6.78 475 11.94 13.60 72.03 06-Nov-19 - 5.72 482 12.78 8.30 63.15