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Population Demographics, Distribution, and Environmental History of Asian Carp in a Great Plains River

Jake Werner University of Nebraska-Lincoln, [email protected]

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Werner, Jake, "Population Demographics, Distribution, and Environmental History of Asian Carp in a Great Plains River" (2020). Dissertations & Theses in Natural Resources. 327. https://digitalcommons.unl.edu/natresdiss/327

This Article is brought to you for free and open access by the Natural Resources, School of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Dissertations & Theses in Natural Resources by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. POPULATION DEMOGRAPHICS, DISTRIBUTION, AND

ENVIRONMENTAL HISTORY OF ASIAN CARP IN A GREAT PLAINS

RIVER

by Jacob P. Werner

A THESIS

Presented to the Faculty of The Graduate College of the University of Nebraska In Partial Fulfillment of Requirements For the Degree of Master of Science

Major: Natural Resource Sciences

Under the Supervision of Professors Martin J. Hamel and Mark A. Pegg

Lincoln, Nebraska

November, 2020 POPULATION DEMOGRAPHICS, DISTRIBUTION, AND

ENVIRONMENTAL HISTORY OF ASIAN CARP IN A GREAT PLAINS

RIVER

Jacob P. Werner, M.S.

University of Nebraska, 2020

Advisers: Martin J. Hamel and Mark A. Pegg

Silver Carp Hypophthalmichthys molitrix, Bighead Carp H. nobilis, and Black

Carp Mylopharyngodon piceus, collectively known as Asian carp, are a group of invasive fishes in the U.S.A. that have garnered much attention over the last couple decades. Most research devoted to this group of fishes has been focused in the Mississippi River basin with little investigation in the Missouri River drainage, particularly in tributary systems.

The Kansas River is a major tributary to the Missouri River that has multiple anthropogenic barriers creating varying levels of connectivity within the Kansas River itself, and with the Missouri River. Information on various life-history traits of Asian carp are needed before a management plan can be formed. Here, we investigated 1) population demographics, 2) distribution with environmental DNA (eDNA), and 3) environmental history using otolith microchemistry of Asian carp in the lower Kansas

River. Silver Carp exhibited spatiotemporal differences in population demographics.

Individuals captured above the lowermost barrier had longer lengths-at-age, longer total lengths, and occurred at lower relative abundance than individuals captured below the barrier. No Silver Carp nor Bighead Carp were detected above the second barrier on the river with physical sampling or with the eDNA assay. However, Black Carp were detected near the confluence with the Missouri River with the eDNA analysis. Otolith microchemistry results indicated the population of Silver Carp in the Kansas River is comprised of predominantly residential individuals. Few carp exhibited natal origin signatures from the Missouri River. Transient individuals within the population exhibited short durations of signatures indicative of the Missouri River, suggesting that movements into the Missouri River are brief. These results highlight the importance of tributary habitat for Asian carp in the Missouri River drainage. Management efforts within the

Kansas River could be an effective means of population control and mitigating secondary introductions. Additionally, management efforts focused in particular reaches of the

Kansas River could affect the greater Missouri River population.

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ACKNOWLEDGEMENTS

I would first like to thank my wife, Christina, for all of her love and support throughout my graduate school experience and in my pursuit of a career in fisheries. Her encouragement and positive attitude pushed me to accomplish my goals. I am truly grateful for her support and understanding, and am thankful to have her by my side throughout this journey.

I would like to thank my advisor, Dr. Marty Hamel, for helping develop my knowledge and skill set in fisheries techniques, and for providing the opportunity to progress as a researcher. I want to thank my committee members, Dr. Mark Pegg, Dr.

Kevin Pope, Dr. Chris Chizinski, and Chris Steffen for providing guidance and feedback throughout my graduate school tenure. Your perspectives, insights, and knowledge kept me headed in the right direction and helped develop my understanding of applied ecology and statistical methods. I want to thank Dr. Samodha Fernando for your guidance with the genetic work.

I would like to thank the Kansas Department of Wildlife, Parks and Tourism for the funding for my project. I want to thank Jeff Koch, Ben Neely, Jeff Conley, Luke

Kowalewski, Connor Ossowski, and Ernesto Flores for their assistance with field sampling. I would also like to thank the folks with the U.S. Fish and Wildlife Service,

Jeremy Hammen, Jason Goeckler, Zach Sanders, Jahn Kallis, and the rest of the crewmembers, for their assistance with field sampling and for providing their specialized gear to help capture Asian carp. I would also like to extend my thanks to our technicians,

Katie Schrag, Charlie DeShazer, and Quade Brees for their hard work in the field and v sustained positivity through long days and the onslaught of flying Silver Carp. This project would not have been possible without the collaboration and hard work from the multiple agencies and numerous people involved.

I want to thank my friend and lab mate, Quintin Dean, for your insightful perspectives, dad jokes, and positivity throughout our graduate school experience. Your hard work and dedication set an example that helped all those around you succeed in their endeavors, and that’s what I appreciates about you. I am thankful for the community provided here at the University of Nebraska – Lincoln. Thank you to Alex Engel, Zach

Horstman, Esther Perisho, Dave Keiter, Lindsay Ohlman, Henry Hansen, Lyndsie

Wszola, Brittany Buchanan, Erin Wood, Mikaela Cherry, and the many others that I was privileged enough to share experiences with along the way. Thank you to all the members of the Pegg, Pope, and Fernando labs for our experiences both in and out of the classrooms.

Lastly, I would like to thank my parents and family for their support throughout my education experience. Their passion for the outdoors and respect for our natural resources greatly influenced my career path into the field of ecology. I would truly not be here without their love and encouragement.

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TABLE OF CONTENTS

CHAPTER 1: GENERAL INTRODUCTION AND STUDY OBJECTIVE….……..…...1 DATA GAP: POPULATION DEMOGRAPHICS……...……….………………..6 DATA GAP: DISTRIBUTION AND UPSTREAM EXTENT…………...…...….7 DATA GAP: ENVIRONMENTAL HISTORY……………………………...…...9 LITERATURE CITED…………………………………………………………..11 CHAPTER 2: POPULATION DEMOGRAPHICS OF SILVER CARP AND BIGHEAD CARP IN THE LOWER KANSAS RIVER, A LARGE TRIBUTARY TO THE MISSOURI RIVER……………………………………………………………..….……19 ABSTRACT……………………………………………………………………..19 INTRODUCTION…………………………………….………………………....20 METHODS……………………………………………………………..………..23 Study Area……………………………………....……………………….23 Field Sampling……………...... …………………………………………24 Laboratory Methods……………………………………………………...25 Data Analysis…………………………………………………………….26 RESULTS………………………………………..………………………………28 DISCUSSION………………………………………………………..…………..30 LITERATURE CITED…………………………………………………………..37 CHAPTER 3: EFFICACY OF ENVIRONMENTAL DNA FOR ASSESSING THE DISTRIBUTION OF INVASIVE ASIAN CARP IN A LARGE TURBID RIVER…….56 ABSTRACT………………………………………………….…………………..56 INTRODUCTION………………………………………………….……………57 METHODS…………………………………………………………………..…..59 Study Area……………………………………………………………….59 Sample Collection………………………………………………………..60 DNA Extraction: Filtration………………………………………………61 DNA Extraction: Centrifugation…………………………………………62 PCR Amplification and Evaluation……………………………………...63 vii

Data Analysis…………………………………………………………….64 RESULTS AND DISCUSSION…………………………..……………………..64 LITERATURE CITED…………………………………………………………..69 CHAPTER 4: ENVIRONMENTAL HISTORY OF SILVER CARP IN THE LOWER KANSAS RIVER, KANSAS: INSIGHT FROM OTOLITH MICROCHEMISTRY ANALYSIS…………………………………………………………………….………..78 ABSTRACT……………………………………………………………………..78 INTRODUCTION………………………………………….……………………79 METHODS…………………………………………………..…………………..81 Study Area………………………………………………………….…....81 Microchemistry Data Collection…………………………………………82 Data Analysis…………………………………………………………….85 RESULTS……………………………………..…………………………………88 Water Chemistry…………………………………………………………88 Otolith Chemistry………………………………………………………...88 DISCUSSION…………………………..………………………………………..91 LITERATURE CITED…………………………………………………………..97 CHAPTER 5: MANAGEMENT AND RESEARCH RECOMMENDATION………..111 MANAGEMENT RECOMMENDATIONS………………….………………..112 Population Control……………………………………………………...112 Monitoring for Asian Carp in Segment 3………………………………114 Movement Deterrents…………………………………………………..116 FUTURE RESEARCH NEEDS…………………………………………..……117 CONCLUSIONS…………………………………………………...…………..120 LITERATURE CITED…………………………………………………………121 APPENDIX A…………………………………………………………………………..124 APPENDIX B…………………………………………………………………………..136 APPENDIX C…………………………………………………………………………..140

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LIST OF TABLES

Table 2-1: Median, first, and third quartiles of catch per unit effort (number/hour) of

Silver Carp with the High Frequency electrofishing (HF), Low Frequency electrofishing

(LF), and the Dozer Trawl in Segments 1, 2, and 3 of the Kansas River.…………….…43

Table 2-2: Mean lengths and ages of adult Silver Carp (>400 mm) captured in Segments

1 and 2 of the Kansas River. Standard deviation provided in parenthesis. Age estimated from lapilli otolith sections.………………………………………………………….…..44

Table 2-3: Total catch of all Silver Carp per segment in 2018 and 2019 from the Kansas

River with all gears deployed...………...………………………………………………..45

Table 2-4: Total catch of all Bighead Carp per segment in 2018 and 2019 from the

Kansas River with all gears deployed……………………………………………………46

Table 3-1: Detection rate (Dr) of Silver Carp, Bighead Carp, and Black Carp at each sampling site in the Kansas River, and from the pilot study (P) near the confluence with the Missouri River using both centrifugation procedures and filtration procedures. Dashes indicate samples from sites that were not processed using those procedures.………...…74

ix

Table 3-2: False negative rate (Fr) for Silver Carp and Bighead Carp in all samples collected in the Kansas River below Bowersock Dam, located in Lawrence, Kansas.….75

Table 3-3: Mean daily discharge (m3 second-1), and mean daily turbidity, measured in

Formazin Nephelometric Units, in the Kansas River on eDNA sample collection days.

Data were collected at the Lake Quivira gaging station located near Edwardsville, KS

(USGS Gage 06892518)…………………………………………………………………76

Table 4-1: Classification matrix for Silver Carp collected from Segment 2 of the Kansas

River and from the Wakarusa River in 2018, and from isolated backwaters of the

Missouri River used to build and test the recursive partitioning tree..…...…………….102

Table 4-2: Percent transient adult and juvenile Silver Carp in Segment 1 and Segment 2 of the Kansas River, and in both segments combined in 2018 and 2019. Total number of fish captured are in parenthesis……..…………………………………………..………103

Table 4-3: Number of transient adult and juvenile Silver Carp captured in the Kansas

River that exhibit 1, 2, and 3, movement events into the Missouri River for each segment.

Natal origins from the Missouri River is the number of transient individuals with

Missouri River Sr:Ca signatures at the nucleus of the otolith……..…………………...104 x

LIST OF FIGURES

Figure 1-1: The lower Kansas River with the three main anthropogenic barriers: the

Johnson County Weir (rkm 27), Bowersock Dam (rkm 83), and the Topeka Weir (rkm

141)……………………………………………………………………………….……...18

Figure 2-1: Kansas River study area where Segment 1 (24 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (61 rkm) is between the Johnson County Weir and Bowersock Dam, and Segment 3 (57 rkm) is between

Bowersock Dam and the Topeka Weir…...……………………………………………...47

Figure 2-2: Mean and 95% confidence intervals of catch per unit effort (CPUE; number/hour) for the Dozer Trawl, High Frequency electrofishing, and Low Frequency electrofishing methods for Segments 1 and 2 of the Kansas River. Error bars represent the

95% confidence intervals for the mean….………………………………………..……...48

Figure 2-3: Length-frequency histograms of Silver Carp captured in Segments 1 and 2 of the Kansas River. Silver Carp were separated into 25 mm length bins and enumerated.

Light grey bars represent Silver Carp captured in 2018 and dark grey bars represent

Silver Carp captured in 2019; 2018 bars are stacked on top of 2019 bars………..……...49

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Figure 2-4: Regressions of log10 transformed lengths and weights of adult Silver Carp

(>400 mm) captured from Segments 1 (light gray) and 2 (dark gray) in 2018 (solid lines) and 2019 (dashed lines) in the Kansas River..…………………………………………...50

Figure 2-5: Length-at-age of Silver Carp in Segments 1 (light gray circles) and 2 (dark gray triangles); data points at each age are offset for clarity. Von-Bertalanffy growth curves were modeled for Segment 1 (light gray line) and Segment 2 (dark gray line).

Growth curves were not offset...…..………………………………………….………….51

Figure 2-6: Predicted lengths-at-age from Von-Bertalanffy growth models for Silver Carp populations in the Illinois River from Grafton, IL to Ottowa IL (red circles and solid line), Segment 1 of the Kansas River (light gray triangles and dotted line), Segment 2 of the Kansas River (dark gray squares and dashed line), and Wabash River from New

Haven, IL to Darwin, IL (blue crosses and dashed line) (Stuck et al. 2015)..…….……..52

Figure 2-7: Length frequency histograms of Silver Carp captured with Dozer Trawl, High

Frequency electrofishing, and Low Frequency electrofishing from the Kansas River in both 2018 and 2019. Silver Carp were separated into 25 mm length bins and enumerated……………………………………………………………………………….53

xii

Figure 2-8: Discharge (blue line), measured in cubic meters per second, and turbidity

(brown line), measured in Formazin Nephelometric Units, in the Kansas River from April of 2018 to October of 2019. Shaded regions represent river conditions during the 2018 sampling season (left) and the 2019 sampling season (right)……………….…………...54

Figure 2-9: Recruitment of Silver Carp captured from Segments 1 (light grey points and solid line) and 2 (dark grey points and dotted line) of the Kansas River. The number of fish in each year class were determined by back-calculating all specimens whose age was determined from sectioned lapilli otoliths in each year of sampling…………………….55

Figure 3-1: Kansas River study area where Segment 1 (24 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (61 rkm) is between the Johnson County Weir and Bowersock Dam, and Segment 3 (57 rkm) is between

Bowersock Dam and the Topeka Weir. White stars indicate water sample collection sites with site numbers next to them………………………………..…………………………77

Figure 4-1: Kansas River study area. Segment 1 (27 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (56 rkm) is between the

Johnson County Weir and Bowersock Dam, and Segment 3 (58 rkm) is between

Bowersock Dam and the Topeka Weir. Red lines indicate the three main barriers on the river and open circles indicate water chemistry sample collection sites………..……...105

xiii

Figure 4-2: Comparison of water microchemistry signatures from the Kansas, Missouri, and Wakarusa Rivers. The horizontal solid line within the box represents the median value, upper and lower limits of the box is quartile ranges, and whiskers are 95% confidence intervals of the median. Points represent data outside of the 95% confidence interval. Median water Sr:Ca and Ba:Ca ratios do not differ between rivers that bear the same letter above the box plot…..……………………………………………...………106

Figure 4-3: Comparison of otolith microchemistry signatures (~ 30µm of data isolated from the edge of the otolith) from Silver Carp captured from Segment 2 of the Kansas

River in 2018, the Wakarusa River in 2018, and isolated backwaters of the Missouri

River. The horizontal solid line within the box represents the median value, upper and lower limits of the box are quartile ranges, and whiskers are 95% confidence intervals of the median. Points represent data outside of the 95% confidence interval. Median otolith

Sr:Ca and Ba:Ca ratios do not differ between rivers that bear the same letter above the box plot……..………...……………………………………………………………..….107

Figure 4-4: Recursive partitioning tree used to classify environmental history of Silver

Carp between the Kansas River and the Missouri River. The tree was pruned at size = 2 with a complexity parameter (cp) of 0.14………………….…………………………...108

xiv

Figure 4-5: Example of ablation data from three Silver Carp captured from the Kansas

River that exhibit trace elemental signatures indicative of the Missouri River. The horizontal line represents the threshold between the Kansas and Missouri Rivers (Sr:Ca =

2,082 µmol/mol) where signatures above the line are from the Missouri River. The shaded region is data that corresponds to the nucleus of the otolith (i.e., natal origins).

Fish number 180208-2 had natal origin signatures from the Missouri River as well as another movement event with signatures from the Missouri River later in life. Fish

180236-18 and 190241-8 both have movement events with signatures from the Missouri

River………………..…………………………………………………………...………109

Figure 4-6: Percent of laser ablation points above 2,082 µmol/mol Sr:Ca threshold for each individual transient fish by total length. Data points greater than 2,082 µmol/mol

Sr:Ca were indicative of Missouri River trace element signatures in lapilli otoliths of

Silver Carp……………………………………………………………………………...110

1 CHAPTER 1: GENERAL INTRODUCTION AND STUDY OBJECTIVES

Introduction and establishment of invasive species are major threats to biodiversity in freshwater systems (Dudgeon et al. 2006; Helfman 2007). Invasive species have changed biological communities in U.S. waterways (Gozlan et al. 2010,

Kolar et al. 2010), leading to the homogenization of aquatic fauna (Kolar et al. 2010).

Invasive species are non-native species that cause ecologic and economic harm if they can establish and expand their range (Gozlan et al. 2010). Silver Carp

Hypophthalmichthys molitrix, Bighead Carp H. nobilis, Grass Carp Ctenopharyngodon idella, and Black Carp Mylopharyngodon piceus, collectively known as Asian carp, are a group of fishes native to eastern Asian that have become widespread, invasive fishes in the United States of America (Conover et al. 2007).

Asian carp were imported into the United States of America in the 1960’s and

1970’s for use in biocontrol practices in aquaculture and wastewater treatment facilities

(Kolar et al. 2005; Conover et al. 2007). Silver Carp and Bighead Carp escaped from these facilities in the 1980’s (Freeze and Henderson 1982) and were able to disperse into

Mississippi River waterways. Subsequently, Silver Carp and Bighead Carp expanded their range to encompass most of the Mississippi River basin and lower Missouri River drainage below Gavin’s Point Dam, South Dakota (Nico et al. 2018a; Nico et al. 2018b).

Silver Carp’s and Bighead Carp’s northernmost extent reaches into North Dakota via the

James River (Hayer et al. 2014). Black Carp were unintentionally imported as a contaminant in a shipment of Grass Carp in the 1970’s, but then were imported intentionally for use in aquaculture facilities in the 1980’s (Conover et al. 2007; Nico and 2

Nielson 2018). Reportedly, they escaped from aquaculture facilities and were able to disperse into the Mississippi River where individual specimens have been documented in the middle and lower Mississippi River, and lower Missouri River downstream of

Jefferson City, Missouri (Nico and Nielson 2018). Grass Carp remain desirable to management agencies as a biocontrol species, although most states permit introduction of triploid specimens only in attempt to prevent natural recruitment (Conover et al. 2007).

Various life-history traits of Asian carp have been investigated in U.S. waters since the invasion of these fishes. Spawning activity of Silver Carp and Bighead Carp is associated with spring flooding events (Conover et al. 2007; DeGrandchamp et al. 2007;

Gibson-Reinemer et al. 2017; Hintz et al. 2017) and occurs in areas of high water velocity (Conover et al. 2007) and turbidity (Deters et al. 2013; Hintz et al. 2017).

Fertilized eggs are semi-buoyant (Conover et al. 2007; Chapman and George 2011) and are thought to require flowing water to stay suspended in the water column (Laird and

Page 1996; Conover et al. 2007). However, there are documented cases of successful reproduction in isolated backwaters of the Illinois River (Garvey et al. 2005). Kolar et al.

(2005) documented a case in which Bighead Carp eggs were discovered in a sediment sample that subsequently hatched and survived for four days. DeGrandchamp et al.

(2007) hypothesized that higher river stage may augment egg and larval survival, but it is not necessary for successful reproduction. Silver Carp and Bighead Carp can also exhibit protracted spawning (Conover et al. 2007), where larger bodied females spawn earlier in the spring and smaller bodied females spawn later in the summer (Hintz et al. 2017). 3

Bighead Carp growth increments peak between ages two and three, reaching lengths over 500 mm by age three (Shrank and Guy 2002). Silver Carp growth rates are more variable, with populations on the invasion front attaining greater lengths at age

(Hayer et al. 2014) and longer total lengths (MacNamara et al. 2016) than established populations (e.g., Williamson and Garvey 2005). Silver Carp and Bighead Carp are capable of long longitudinal movements, which are typically associated with a rise in river stage (Peters et al. 2006; DeGrandchamp et al. 2008; Coulter et al. 2016). Some

Silver Carp have been documented moving as far as 64 km in a day (DeGrandchamp et al. 2008) and a single Bighead Carp was documented moving 163 km over a five week period (Peters et al. 2006). During low summer flows Silver Carp and Bighead Carp occupy side channel habitats more frequently than main channel and backwater habitats

(DeGrandchamp et al. 2008), while also showing a preference for areas with slower moving water over static areas (Calkins et al. 2012). Juvenile Silver Carp and Bighead

Carp tend to be found in areas of lower flow (Haupt and Phelps 2016), such as areas behind wing dikes or tributaries (Kolar et al. 2005; Williamson and Garvey 2005).

Competitive interactions between Asian carp and native biota have also been investigated. Silver Carp and Bighead Carp are filter feeders with broad diets mainly consisting of zooplankton and phytoplankton (Conover et al. 2007; Sampson et al. 2009).

These two carp species directly compete with native filter feeders such as Gizzard Shad

Dorosoma cepedianum and Bigmouth Buffalo Ictiobus cyprinellus (Irons et al. 2007;

Sampson et al. 2009). Competitive interactions have also been documented with age-0

Paddlefish Polyodon spathula (Schrank et al. 2003), although there is little dietary 4 overlap between adult Paddlefish and adult Silver Carp and Bighead Carp (Sampson et al.

2009). Zooplankton community composition and structure can be altered by Silver Carp and Bighead Carp (Cooke et al. 2009; Sass et al. 2014; DeBoer et al. 2017). Altered zooplankton communities can have an effect on other fish species by limiting food resources to the larval stage of these fishes before their ontogenetic dietary shifts (Sass et al. 2014; Solomon et al. 2016; Chick et al. 2020). Diets of adult Black Carp are predominantly comprised of snails and mollusks (Nico et al. 2005; Nico and Nielson

2018). Establishment of Black Carp is concerning because of their impact on native freshwater mussels (Nico et al. 2005), which are already disproportionally imperiled compared to other taxa in the U.S.A. and Canada (Williams et al. 1993). Black Carp are capable of attaining large body sizes (Nico and Nielson 2018), allowing them to prey on the largest native mussels.

Silver Carp pose a threat to boaters as they have a propensity to jump out of the water when disturbed, potentially causing bodily harm and property damage (Conover et al. 2007). Threats such as this could alter recreational experiences and activity patterns of boaters in areas infested with these fish (Wittman et al. 2015). Additionally, competition between Asian carps and popular sportfish could cause a decline in the quality of the sportfish community thereby reducing use of the fishery. Likewise, establishment of

Asian carp can also compromise commercial fisheries (Connover et al. 2007). As such, there is concern about Asian carps establishing in the Laurentian Great Lakes and depleting crucial resources (Cooke and Hill 2010). Management of Asian carps is direly needed to negate ecological and economic impacts in U.S. waters. 5

Management of invasive species often focuses on eradication and population control (Kolar et al. 2010). Several methods of eradication have been attempted, such as mechanical removals and rotenone treatments despite little success. Once a species has become established in open river systems, eradication is nearly impossible and management objectives shift to population control (Kolar et al. 2010). Life-history traits, species distribution, and other biological information should be collected before a population control plan can be formulated and implemented (Conover et al. 2007; Kolar et al. 2010). Most research concerning various life-history traits of Asian carp has been conducted in the Mississippi River basin with little research coming from the Missouri

River drainage. Information on life-history traits and requirements are vital to mitigate

Asian carps’ advance up the Missouri River basin, including in tributaries like the Kansas

River.

The Kansas River, Kansas, is a large tributary to the Missouri River. Young-of- year (YOY) Silver Carp were first documented in the Kansas River in 2010 (Mosher

2014b) and Bighead Carp were first detected in 1993 (Mosher 2014a). Previous assessments on the Kansas River from 2005 to 2007 failed to capture any species of

Asian carp (Eitzmann and Paukert 2010; White et al. 2010). However, Wanner and

Klumb (2009) captured Silver Carp in the Missouri River upstream of the Kansas-

Missouri confluence in 2003, indicating Silver Carp were likely present in the Kansas

River, but at low abundance. Black Carp have never been documented in the Kansas

River. The furthest upstream a Black Carp specimen has been found in the Missouri 6

River is just east of Jefferson City, Missouri (Nico and Nielsen 2018), about 350 river kilometers (rkm) downstream of the Kansas-Missouri confluence.

The Kansas River has multiple anthropogenic barriers throughout its lower half

(Figure 1-1); the Johnson County Weir at rkm 27, Bowersock Dam at rkm 83, and the

Topeka Weir at rkm 141. Bowersock Dam is a hydropower dam that is classified as a low-head dam (Quist and Guy 1999) and has been suggested to impede upstream longitudinal movements of fishes in this river (Eitzmann et al. 2007; Dean 2020). Silver

Carp and Bighead Carp have been documented as far upstream as Bowersock Dam, but

Silver Carp have never been documented upstream of this barrier. A few Bighead Carp were able to traverse the dam during a flooding event in 1993 (peak discharge: 2,481 m3 second-1; Quist et al. 1999) but presumably have not established a reproducing population upstream of the dam (C. Steffen, Kansas Department of Wildlife, Parks and Tourism, personal communications). Currently, little is known about Asian carp in the Kansas

River and data gaps concerning population demographics, distribution, and environmental history of Asian carp within this river system need to be addressed. Such data will lead to insights as to how Asian carp are functioning in a tributary to a large river and will establish a basis for a management plan.

Data gap: population demographics

Most research conducted on Silver Carp and Bighead Carp in North America has focused on life-history attributes such as movement (Peters et al. 2006; DeGrandchamp et al. 2008; Coulter et al. 2016), reproduction (DeGrandchamp et al. 2007; Deters et al.

2013; Hintz et al. 2017), competitive interactions (Schrank et al. 2003; Irons et al. 2007; 7

DeBoer et al. 2018; Chick et al 2020), distribution (Jerde et al. 2011; Jerde et al. 2013), and natal origins (Norman and Whitledge 2015; Whitledge et al 2018). However, management agencies also recognize the value in basic population demographic data to manage these invasive fishes (e.g., Conover et al. 2007; Hayer et al. 2014). Metrics such as condition, relative abundance, size structure, and age and growth provide managers information as to the best management action for that particular population (Kolar et al.

2010). These metrics also provide a basis to assess the efficacy of management strategies

(Allen and Hightower 2010) by establishing a base line. Additionally, we can assess how

Silver Carp and Bighead Carp respond to varying biotic or abiotic factors. Therefore, our first objective was to assess population demographics (i.e., age and growth, relative abundance, condition, and size structure) of Silver Carp and Bighead Carp in the Kansas

River.

Data gap: distribution and upstream extent

Preventing range expansion of Asian carp is imperative to minimize their detrimental ecologic and economic impacts (Conover et al. 2007). One method of range expansion is through secondary introductions that occurs when carp spread from infested to non-infested waters (Vander Zanden and Olden 2008). Mitigating secondary introductions is important for ensuring ecosystem resilience (Vander Zanden and Olden

2008). Information on the current distribution of Asian carp and on waters that are most vulnerable to the spread of these fishes is imperative to prevent secondary introductions.

Vander Zanden and Olden (2008) assessed vulnerability using three discrete stages: 1) arrival, 2) establishment, and 3) impact. Arrival is whether reproductively viable 8 individuals can reach new systems. Establishment is if the species can successfully reproduce in the system. Impact is whether the species will cause ecologic harm in new systems (Vander Zanden and Olden 2008).

Arrival of Asian carp into non-infested waters can be facilitated by large river systems. The large spatial scales and connectivity to a multitude of tributaries within these systems (i.e., Mississippi and Missouri Rivers) provides a pathway for Asian carp to distribute (Ratcliff et al. 2014). However, most large river systems – such as the

Missouri River – tend to have high levels of anthropogenic alteration (e.g., Galat et al.

1998). Dams and diversions are a common source of longitudinal fragmentation (Paukert and Galat 2010) that can impede upstream movements (Porto et al.1999) and restrict ranges (Jelks et al. 2008). Silver Carp and Bighead Carp have been documented traversing some anthropogenic barriers (Moy et al. 2011; Jerde et al. 2013; Tripp et al.

2014) and expanding into new waters. Additional research understanding Asian carp movement impediments is warranted.

Bowersock Dam, in Lawrence, Kansas, is presumed to be a barrier to upstream movement of riverine fishes in the Kansas River (e.g., Eitzmann et al. 2007; Dean 2020)

(Figure 1-1). Although Bighead Carp were able to traverse this barrier during a flooding event in the early 1990’s, the dam has since undergone major alterations beginning in

2011. A new powerhouse was constructed on the North side of the river where the old spillway was previously located. This construction created a larger barrier to upstream fish movement in that section. The dam top height did not change during construction and is currently at 808 National Geodetic Vertical Datum (NGVD). An inflatable dam 9 system can increase the dam height to between 813 and 814 NGVD during lower flows.

After the river exceeds approximately 566 m3 second-1 the dam top structure has to be deflated (Hill-Nelson, The Bowersock Mill and Power Company, personal communication).

Silver Carp have never been documented upstream of Bowersock Dam. However,

Silver Carp and Bighead Carp are difficult to detect with conventional gears at low population densities (Jerde et al. 2011; Jerde et al. 2013). Black Carp are benthic oriented and difficult to detect; therefore, their true distribution is most likely greater than what is presently known (Nico and Jelks 2011). Environmental DNA (eDNA) is a useful tool for detecting rare fish or fish at low abundances (Jerde et al. 2011; Laramie et al. 2015;

O’Sullivan et al. 2020) and has been used to monitor for Asian carp in vulnerable waters

(Jerde et al. 2011; Jerde et al. 2013). Therefore, our second objective was to provide insight into the distribution of Silver Carp and Bighead Carp in the lower Kansas River with an eDNA assay. These data will help determine if these fish are present upstream of

Bowersock Dam. Additionally, we used eDNA to monitor for Black Carp in the lower

Kansas River.

Data gap: environmental history

Silver Carp can exhibit dichotomy in individual based movement patterns where some individuals in a given population are transient and others are residents (Coulter et al. 2016; Prechtel et al. 2018) Discerning the proportion of individuals that are transient and resident can provide insight for management action. For example, timing removal efforts to coincide with periods of reduced movement distances (i.e., summer and early 10 fall) could be effective means for removing transient individuals (Prechtel et al. 2018).

Additionally, populations comprised of predominantly resident individuals would be ideal for removal efforts.

Analysis of stable isotopes and trace element composition of otoliths as natural tags have been used to retrospectively identify environmental history of riverine fishes

(Zeigler and Whitledge 2010). Strontium (Sr) and Barium (Ba) – in ratios to Calcium

(Ca) - are two common trace elements used in microchemistry analysis (Whitldedge et al.

2007; Zeigler and Whitledge 2010; Crook et al. 2013; Norman and Whitledge 2015;

Carlson et al. 2016; Whitledge et al. 2018). For example, Sr:Ca ratios were used to identify natal origins of Silver Carp captured in the Illinois River to the Illinois River itself, the Missouri River, or the middle Mississippi River (Norman and Whitledge 2015).

Additionally, Sr:Ca ratios were used to identify origins of Silver Carp captured in urban

Chicago fishing ponds to the Illinois River (Love et al. 2019). Therefore, our third objective was to identify environmental history of Silver Carp captured throughout the

Kansas River using otolith microchemistry. These data provided insight into source-sink dynamics within the Missouri River basin and in understanding the role a large tributary to the Missouri River plays in various life-history attributes of this invasive species. 11

Literature Cited Allen, M. S., and J. E. Hightower. 2010. Fish population dynamics: mortality, growth and recruitment. Pages 43-80 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Calkins, H. A., S. J. Tripp, and J. E. Garvey. 2012. Linking silver carp habitat selection to flow and phytoplankton in the Mississippi River. Biological Invasions 14:949- 958. Carlson, A. K., M. J. Fincel, and B. D. S. Graeb. 2016. Otolith microchemistry reveals natal origins of walleyes in Missouri river reservoirs. North American Journal of Fisheries Management 36:341-350. Chapman, D. C. and A. E. George, 2011. Developmental rate and behavior of early life stages of bighead carp and silver carp. U.S. Geological Survey Scientific Investigations Report 2011–5076, 62 p. Chick, J. H., D. K. Gibson-Reinemer, L. Soeken-Gittinger, and A. F. Casper. 2020. Invasive Silver Carp is empirically linked to declines of native sportfish in the Upper Mississippi River system. Biological Invasions 22:723-734. Conover, G., R. Simmons, and M. Whalen. 2007. Management and Control plan for Bighead, Black, Grass, and Silver Carps in the United States. Asian carp Working Group, Aquatic Nuisance Species Task Force, Washington D.C. 223 pp. Cooke, S. L. and W. R. Hill. 2010. Can filter-feeding Asian carp invade the Laurentian Great lakes? A bioenergetics modelling exercise. Freshwater Biology 55:2138- 2152. Cooke, S. L., W. R. Hill, and K. P. Meyer. 2009. Feeding at different plankton densities alters invasive Bighead Carp (Hypophthalmichthys nobilis) growth and zooplankton species composition. Hydrobiologia 625:185-193. Coulter, A. A., E. J. Bailey, D. Keller, and R. R. Goforth. 2016. Invasive Silver Carp movement patterns in the predominantly free-flowing Wabash River (Indiana, USA). Biological Invasions 18:471-485. Crook, D. A., J. I. Macdonald, D .G. McNeil, D. M. Gilligan, M. Asmus, R. Maas, and J. Woodhead. 2013. Recruitment sources and dispersal of an invasive fish in a large river system as revealed by otolith chemistry analysis. Canadian Journal of Fisheries and Aquatic Sciences 70:953-963. Dean, Q. J. 2020. Population characteristics and movement of Blue Catfish in the Kansas River. Master’s thesis. University of Nebraska, Lincoln. 12

DeBoer, J. A., A. M . Anderson, and A. F. Casper. 2018. Multi-trophic response to invasive Silver Carp (Hypophthalmichthys molitrix) in a large floodplain river. Freshwater Biology 63:597-611. DeGrandchamp, K. L., J. E. Garvey, and R. E. Colombo. 2008. Movement and habitat selection by invasive Asian carp in a large river. Transactions of the American Fisheries Society 137:45-56. DeGrandchamp, K. L., J. E. Garvey, and L. A. Csoboth. 2007. Linking adult reproduction and larval density of invasive carp in8 a large river. Transactions of the American Fisheries Society 136:1327-1334. Deters, J. E., D. C. Chapman, and B. McElroy. 2013. Location and timing of Asian carp spawning in the lower Missouri river. Environmental Biology of Fishes 96:617- 629. Dudgeon, D., A. H. Arthington, M. O Gessner, Z. I. Kawabata, D. J. Knowler, C Lévêque, R. J. Naiman, A. H. Prieur-Richard, D. Soto, M. L. J. Stiassny, and C. A. Sullivan. 2006. Freshwater biodiversity: importance, threats, status, and conservation challenges. Biological Review 81:163-182. Eitzmann, J. L., A. S. Makinster, and C. P. Paukert. 2007. Distribution and growth of blue sucker in a great plains river, USA. Fisheries Management and Ecology 14:255-262. Eitzmann, J. L. and C. P. Paukert. 2010. Longitudinal differences in habitat complexity and fish assemblage structure of a great plains river. The American Midland Naturalist 163:14-32. Freeze, M. and S. Henderson. 1983. Distribution and Status of the Bighead Carp and Silver Carp in Arkansas. North American Journal of Fisheries Management 2:197-200. Galat, D. L., L. H. Fredrickson, D. D. Humburg, K. J. Bataille, J. R. Bodie, J. Dohrenwend, G. T. Gelwicks, J. E. Havel, D. L. Helmers, J. B. Hooker, J. R. Jones, M. F. Knowlton, J. Kubisiak, J. Mazourek, A. C. McColpin, R. B. Renken, and R. D. Semlitsch. 1998. Flooding to restore connectivity of regulated, large- river wetlands. BioSciene 48:721-733. Garvey, J. E., R. Brooks, M. Eichholz, and J. Chick. 2005. Swan lake habitat rehabilitation and enhancement project: post-project monitoring of fish movement, fish community, waterfowl, water quality, vegetation, and invertebrates. Year 1 Summary to US Army Corps of Engineers, St. Louis District. 135 pages. Gibson-Reinemer, D. K., L. E. Solomon, R. M. Pendleton, J. H. Chick, and A. F. Casper. 2017. Hydrology controls recruitment of two invasive cyprinids: bighead carp reproduction in a navigable large river. PeerJ 5:e364. 13

Gozlan, R. E., J. R. Britton, I. Cowx, and G. H. Copp. 2010. Current knowledge on non- native freshwater fish introductions. Journal of Fish Biology 76:751-786. Haupt, K. J., and Q. E. Phelps. 2016. Mesohabitat association in the Mississippi River Basin: a long-term study on the catch rates and physical habitat association of juvenile Silver Carp and two native planktivores. Aquatic Invasions 11:93-99. Hayer, C.-A., J. J. Breeggemann, R. A. Klumb, B. D. S. Graeb, and K. N. Bertrand. 2014. Population characteristics of Bighead and Silver Carp on the Northwestern front of their North American invasion. Aquatic Invasions 9:289-303. Helfman, G. S. 2007. Fish conservation: a guide to understanding and restoring global aquatic biodiversity and fishery resources. Island Press, Washington, D.C. Hintz, W. D., D. C. Glover, B. C. Szynkowski, and J. E. Garvey. 2017. Spatiotemporal reproduction and larval habitat associations of nonnative Silver Carp and Bighead Carp. Transactions of the American Fisheries Society 146:422-431. Irons, K. S., G. G. Sass, M. A. McClelland, and J. D. Stafford. 2007. Reduced condition factors for two native fish species coincident with invasion of non-native Asian carps in the Illinois River, U.S.A. Is this evidence for competition and reduced fitness? Journal of Fish Biology 71:258-273. Jelks, H. L., S. J. Walsh, N. M. Burkhead, S. Contreras-Balderas, E. Diaz-Pardo, D. A. Hendrickson, J. Lyons, N. E. Mandrak, F. McCormick, J. S. Nelson, S. P. Platania, B. A. Porter, C. B. Renaud, J. J. Schmitter-Soto, E. B. Taylor, and M. L Warren Jr. 2008. Conservation status of imperiled North American freshwater and diadromous fishes. Fisheries 33:372-407. Jerde, C. L., W. L. Chadderton, A. R. Mahon, M. A. Renshaw, J. Corush, M. L. Budney, S. Mysorekar, and D. M. Lodge. 2013. Detection of Asian carp DNA as part of a Great-Lakes basin wide surveillance program. Canadian Journal of Fisheries and Aquatic Sciences 70:522-526. Jerde, C. L., A. R. Mahon, W. L. Chadderton, and D. M. Lodge. 2011. “Sight-unseen” detection of rare aquatic species using environmental DNA. Conservation Letters 4:150-157. Kolar, C. S., D. C. Chapman, W. R. Courtenay Jr., C. M. Housel, and J. D. Williams. 2005. Asian carps of the Genus Hypophthalmichthys (Pisces, Cyprinidae) – A biological synopsis and environmental risk assessment. National Invasive Species Council materials. 5. Kolar, C. S., W. R. Courtenay, and L. G. Nico. 2010. Managing undesired and invading fishes. Pages 213-259 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. 14

Laird, C. A. and L. M. Page. 1996. Non-native fishes inhabiting the streams and lakes of Illinois. Illinois Natural History Survey Bulletin 35:1-51. Laramie, M. B., D. S. Pilliod, and C. S. Goldberg. 2015. Characterizing the distribution of an endangered salmonid using environmental DNA analysis. Biological Conservation 183:29-37. MacNamara, R., D. Glover, J. E. Garvey, W. Bouska, and K. Irons. 2016. Bigheaded carps (Hpophthalmichthys spp.) at the edge of their invaded range: using hydroacoustics to assess population parameters and the efficacy of harvest as a control strategy in a large North American river. Biological Invasions 18:3293- 3307. Mosher, T. D. 2014a. Bighead Carp, Hypophthalmichthys nobilis Richardson 1845. Pages 166-168 in Kansas Fishes Committee. Kansas fishes. University Press of Kansas, Lawrence. Mosher, T. D. 2014b. Silver Carp, Hypophthalmichthys molitrix Richardson 1845. Pages 166-167 in Kansas Fishes Committee. Kansas fishes. University Press of Kansas, Lawrence. Moy, P. B., I. Polls, and J. M. Dettmers. 2011, The Chicago sanitary and ship canal aquatic nuisance species dispersal barrier. Pages 121-138 in D. C. Chapman and M. H. Hoff, editors. Invasive Carps in North America. American Fisheries Society, Symposium 74, Bethesda, Maryland Nico, L. G. and H. J. Jelks. 2011. The black carp in north America: an update. American Fisheries Society Symposium 74:89-104. Nico, L. G., and M. E. Neilson. 2018. Mylopharyngodon piceus (Richardson, 1846); USGS Nonindigenous Aquatic Species Database, Gainesville, FL. Available: https://nas.er.usgs.gov/queries/FactSheet.aspx?speciesID=573. Revision date: 8/6/2018. Nico, L. G., P. Fuller, and J. Li, 2018a, Hypophthalmichthys molitrix (Valenciennes in Cuvier and Valenciennes, 1844): U.S. Geological Survey, Nonindigenous Aquatic Species Database, Gainesville, FL, https://nas.er.usgs.gov/queries/factsheet.aspx?speciesID=549, Revision Date: 10/25/2018, Peer Review Date: 4/1/2016. Nico, L. G., P. Fuller, and J. Li, 2018b, Hypophthalmichthys nobilis (Richardson, 1845): U.S. Geological Survey, Nonindigenous Aquatic Species Database, Gainesville, FL, https://nas.er.usgs.gov/queries/factsheet.aspx?SpeciesID=551, Revision Date: 10/25/2018, Peer Review Date: 4/1/2016, Access Date: 12/15/2018. Nico, L. G., J. D. Williams, and H. L. Jelks. 2005. Black Carp: biological synopsis and risk assessment of an introduced fish. American Fisheries Society, Special Publication 32, Bethesda, Maryland. 15

Norman, J. D., and G. W. Whitledge. 2015. Recruitment sources of invasive bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) inhabiting the Illinois river. Biological Invasions 17:2999-3014. O’Sullivan, A. M., K. M. Samways, A. Perreault, C. Hernandez, M. D. Gautreau, R. A. Curry, and L. Bernatchez. 2020. Space invaders: searching for invasive Smallmouth Bass (Micropterus dolomieu) in a renowned Atlantic Salmon (Salmo salar) river. Ecology and Evolution 00:1-9. Paukert, C. A. and D. L. Galat. 2010. Warmwater rivers. Pages 699-736 in W.A Hubert and M.C Quist, editors. Inland Fisheries Management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Peters, L. M., M. A. Pegg, and U. L. Reinhardt. 2006. Movement of adult radio-tagged Bighead Carp in the Illinois River. Transactions of the American Fisheries Society 135:1205-1212. Porto, L. M., R. L. McLaughlin, and D. L. G. Noakes. 1999. Low-head barrier dams restrict movements of fishes in two Lake Ontario streams. North American Journal of Fisheries Management 19:1028-1036. Prechtel, A. R., A. A. Coulter, L. Etchison, P. R. Jackson, and R. R. Goforth. 2018. Rang estimates and habitat use of invasive Silver Carp (Hypophthalmichthys molitrix): evidence of sedentary and mobile individuals. Hydrobiologia 805:203-218. Quist, M. C., and C. S. Guy. 1999. Spatial variation in population characteristics of the Shovelnose Sturgeon in the Kansas River. The Prairie Naturalist 31:65-74. Quist, M. C., J. S. Tillma, M. N. Burlingame, and C. S. Guy. 1999. Overwinter habitat use of shovelnose sturgeon in the Kansas River. Transactions of the American Fisheries Society 128:522-527. Ratcliffe, E. N., E. J. Gittinger, T. M. O’Hara, and B. S. Ickes. 2014. Long term resource monitoring program procedures: fish monitoring, 2nd edition. A program report submitted to the U.S. army corps of engineers’ upper Mississippi River restoration-environmental management program. June 2014. Program report LTRMP 2014-P001. 88 pp. Including apendages A-G. Sampson, S. J., J. H. Chick, and M. A. Pegg. 2009. Diet overlap among two Asian carp and three native fishes in backwater lakes on the Illinois and Mississippi rivers. Biological Invasions 11:483-496. Sass, G. S., C. Hinz, A. C. Erickson, N. N. McClelland, M. A. McClelland, and J. M. Epifanio. 2014. Invasive bighead and silver carp effects on zooplankton communities in the Illinois River, Illinois USA. Journal of Great Lakes Research 40:911-921.

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Schrank, S. J., P. J. Braaten, and C. S. Guy. 2003. Spatiotemporal variation in density of larval Bighead Carp in the lower Missouri River. Transactions of the American Fisheries Society 130:809-814.

Schrank, S. J. and C. S. Guy. 2002. Age, growth, and gonadal characteristics of adult bighead carp, Hypophthalmichthys nobilis, in the lower Missouri River. Environmental Biology of Fishes. 64:443-450.

Solomon, L. E., R. M. Pendleton, J. H. Chick, and A. F. Casper. 2016. Long-term changes in fish community structure in relation to the establishment of Asian carp in a large floodplain river. Biological Invasions 18:2883-2895. Spurgeon, J. J., M. A. Pegg, and N. M. Halden. 2018. Mixed-origins of channel catfish in a large-river tributary. Fisheries Research. 198: 195-202. Stuck, J. G., A. P. Porreca, D. H. Wahl, and R. E. Colombo. 2015. Contrasting population demographics of invasive silver carp between and impounded and free-flowing river. North American Journal of Fisheries Management 35:114-122. Tripp, S., R. Brooks, D. Herzog, and J. Garvey. 2014. Patterns of fish passage in the upper Mississippi River. River Research and Application 30:1056-1064. Vander Zanden, J. M. and J. D. Olden. 2008. A management framework for preventing the secondary spread of aquatic invasive species. Canadian Journal of Fisheries and Aquatic Sciences 65:1512-1522. Wanner, G. A., and R. A. Klumb. 2009. Asian carp in the Missouri river: analysis from multiple Missouri river habitat and fisheries programs. National Invasive Species Council materials. Paper 10. White, K., J. Gerken, C. P. Paukert, and A. Makinster. 2010. Fish community structure in natural and engineered habitats in the Kansas River. River Research and Applications 26:797-805. Whitledge, G. W., B. M. Johnson, P. J. Martinez, and A. M. Martinez. 2007. Sources of nonnative centrarchids in the upper Colorado River revealed by stable isotope and microchemical analyses of otoliths. Transactions of the American Fisheries Society 136: 1263-1275. Whitledge, G. W., B. Knights, J. Vallazza, J. Larson, M. J. Weber, J. T. Lamer, Q. E. Phelps, and J. D. Norman. 2018. Identification of Bighead Carp and Silver Carp early-life environments and inferring Lock and Dam 19 passage in the upper Mississippi River: insights from otolith chemistry. Biological Invasions.

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Williamson, C. J., and J. E. Garvey. 2005. Growth, fecundity, and diets of newly established silver carp in the middle Mississippi river. Transactions of the American Fisheries Society 134:1423-1430. Williams, J. D., M. L. Warren, K. S. Cummings, J. L. Harris, and R. J. Neves. 1993. Conservation status of freshwater mussels of the United States and Canada. Fisheries 18:6-22. Wittmann, M. E., R. M. Cooke, J. D. Rothlisberger, E. S. Rutherford, H. Zhang, D. M. Mason, and D. M. Lodge. 2015. Use of structured expert judgement to forecasr invasions by Bighead and Silver Carp in Lake Erie. Conservation Biology 29:187- 197. Zeigler, J. M. and G. W. Whitledge. 2010. Assessment of otolith chemistry for identifying source environment of fishes in the lower Illinois river, Illinois. Hydrobiologia 638:109-119.

18

Figure 1-1: The lower Kansas River with the three main anthropogenic barriers: the

Johnson County Weir (rkm 27), Bowersock Dam (rkm 83), and the Topeka Weir (rkm

141). 19

CHAPTER 2: POPULATION DEMOGRAPHICS OF SILVER CARP AND BIGHEAD CARP IN THE LOWER KANSAS RIVER; A LARGE TRIBUTARY TO THE MISSOURI RIVER

J. P. WERNER*, Q. J. DEAN, M. A. PEGG, and M. J. HAMEL

Abstract Silver Carp Hypophthalmichthys molitrix and Bighead Carp H. nobilis, collectively known as bigheaded carp, have expanded their range to encompass most of the Mississippi River basin, including much of the Missouri River. However, there is a paucity of information on bigheaded carp in the Missouri River basin, especially in the tributaries. Little is known about how carp function in these tributaries or how connectivity with a mainstem river can influence population demographics within either system. The Kansas River is a large tributary to the Missouri River and has multiple anthropogenic barriers (e.g., hydropower dams and water diversion weirs) creating varying levels of connectivity within the system, as well as with the Missouri River.

These varying levels of connectivity provide a unique opportunity to analyze how river connectivity – or lack thereof – can influence bigheaded carp population demographics across the landscape. We collected Silver Carp (n=1,612) and Bighead Carp (n=39) from above and below two barriers in the Kansas River using a suite of gears in the summers of 2018 and 2019. No Silver Carp or Bighead Carp were captured above the upstream- most of the two barriers (hydropower dam at river kilometer (rkm) 83), but both species were found above a water diversion weir at rkm 27. Adults captured above the weir occurred in lower densities and had greater growth rates (F1:583 = 21.69, P < 0.001) 20 reaching longer lengths at age. Juvenile Silver Carp were scarce above the weir but were abundant below. Length-weight relationships of Silver Carp were different between years both above (F1; 4554 = 6.753, P < 0.01) and below (F1; 668 = 16.82, P < 0.001) the diversion weir, which relate to drastic differences in the mean annual discharge that occurred over the course of this study. Connectivity within the Kansas River via the water diversion weir appears to influence size structure and density of Silver carp above and below the weir. Lack of juveniles above the weir indicate reproduction may be limited in this reach.

River conditions below the weir in the Kansas River may be more suitable for rearing juvenile Silver Carp.

Introduction

Introduction of non-native species to U.S. waterways have changed biological communities (Gozlan et al. 2010; Kolar et al. 2010), often in an irreversible way (Kolar and Lodge 2002). Silver Carp Hypophthalmichthys molitrix and Bighead Carp H. nobilis, collectively known as bigheaded carp, were introduced into Mississippi waterways in the

1970’s (Freeze and Henderson 1982; Conover et al. 2007). They have expanded their distribution to include much of the Missouri River drainage and now range as far north as

North Dakota (Hayer et al. 2014). These invasive carp can impact native filter feeders such as Gizzard Shad Dorosoma cepedianum and Bigmouth Buffalo Ictiobus cyprinellus through direct competition (Irons et al. 2007; Sampson et al. 2009). Additionally, bigheaded carp can facilitate shifts in zooplankton community structure and composition

(Sass et al. 2014; DeBoer et al. 2017). This shift in zooplankton community structure can 21 impact other fish species by limiting food availability for larval fishes that forage on plankton before their ontogenetic dietary shift (Sass et al. 2014; Solomon et al. 2016;

Chick et al. 2020). Various life-history aspects of bigheaded carp have been investigated throughout the Mississippi River drainage to inform management decisions

(Degrandchamp et al. 2008; Sampson et al. 2009; Jerde et al. 2013; Norman and

Whitledge 2015; Stuck et al. 2015); however, there is a paucity of information on bigheaded carp in the Missouri River, especially in its tributary systems.

Tributaries are crucial in the life cycle of many riverine fishes (Neely et al. 2009;

Bottcher et. al 2013; Brönmark et al. 2014; Hamel et al. 2014) and may be important for certain life-history attributes of bigheaded carp. For example, Silver Carp populations within the Mississippi River drainage can be comprised of individuals with natal origins from multiple systems, including tributaries (Norman and Whitledge 2015). Tributaries in the Missouri River may be important because the mainstem has undergone a multitude of alterations (e.g., installation of dams, wing dikes, and rip-rap). These alterations have limited lateral connectivity with the river floodplain and increased mean velocity and mean channel depth (Galat et al. 1998; Pegg et al. 2003; Steffenson and Mestl 2016).

Consequently, limited optimal habitat remains for bigheaded carp in the Missouri River as they tend to prefer waters with lower velocities (Kolar et al. 2005; Calkins et al. 2012).

Missouri River tributaries, and limited habitat behind wing dikes, may provide important refuge habitat for bigheaded carp avoiding the swift flows in the main channel of the

Missouri River (Kolar et al. 2005). Understanding how bigheaded carp function in tributaries to the Missouri River is essential in developing regional and basin-wide 22 control plans. One such tributary, the Kansas River, has an established population of bigheaded carp that may provide insight on the importance of connectivity between the mainstem and tributaries in the Missouri River basin.

Bighead Carp were first documented in the Kansas River in 1993 (Mosher

2014a), and young-of-year (YOY) Silver Carp were first documented in 2010 (Mosher

2014b). Adults of both species have been found upstream as far as Lawrence, Kansas

(rkm 83). Previous assessments in the Kansas River failed to detect Silver Carp from

2005 to 2007 (Eitzmann and Paukert 2010; White et al. 2010); however, Silver Carp were known to be in the Missouri River upstream of the Kansas-Missouri confluence at this time (Wanner and Klumb 2009), so it is likely they were present in the Kansas River at low densities.

The Kansas River has multiple anthropogenic barriers creating varying levels of connectivity within the Kansas River as well as with the Missouri River. Connectivity between river reaches and with the Missouri River is not always completely blocked pending water levels (Dean 2020). This variation provides a unique opportunity to investigate how anthropogenic fragmentation within a tributary and connectivity with a main stem river can influence population demographics of bigheaded carp.

Investigating population demographics in the Kansas River is paramount for developing and implementing a control plan. Information on how bigheaded carp are functioning in this system provides insight for appropriate management strategies (Kolar et al. 2010) by identifying where and when control efforts may be most effective.

Additionally, assessment of these metrics (i.e., age and growth, size structure, relative 23 abundance, and condition) enables managers to detect changes in the population in response to management action (Allen and Hightower 2010) or varying abiotic factors such as anthropogenic fragmentation. Our objective was to compare population demographics (i.e., relative abundance, condition, size structure, and age and growth) of bigheaded carp captured above and below two of the main barriers in the Kansas River.

The use of multiple gears to capture bigheaded carp also provided insight as to the most appropriate sampling gear for this system. These data will help managers understand how bigheaded carp are functioning in tributaries as well as how connectivity with a main stem river can influence population demographics.

Methods

Study Area

The Kansas River begins at the confluence of the Smoky Hill and Republican

Rivers (Quist and Guy 1999) and flows Easterly for 274 kilometers to its confluence with the Missouri River in Kansas City, Kansas (Figure 2-1). Discharge within the basin is controlled by 18 federal reservoirs and over 13,000 small impoundments. Bowersock

Dam at rkm 83 near Lawrence, Kansas is the only dam on the Kansas River and is classified as a low-head dam (Quist et al. 1999; Makinster and Paukert 2008). There are two water diversion weirs between Topeka, Kansas, and the confluence with the Missouri

River: the Johnson County Weir at rkm 27 (Eitzmann and Paukert 2010) and the Topeka weir at rkm 141.

We separated the study area into three segments; Segment 1 (27 rkm) is from the confluence with the Missouri River to the Johnson County Weir, Segment 2 (56 rkm) is 24 from the Johnson County Weir to Bowersock Dam, and Segment 3 (58 rkm) is from

Bowersock Dam to the Topeka Weir. Each segment was further stratified into sampling sites to ensure sampling effort was distributed throughout each segment. Sampling sites were determined as sections of the river between access locations or between access locations and the three main barriers on the river.

Field Sampling

We collected bigheaded carp in May through August 2018 and 2019 using a suite of gears. We used boat-mounted electrofishers (Irons et al. 2007; Bouska et al. 2017), an electrified dozer trawl (Hammen et al. 2019), mini-fyke nets (Collins et al. 2017; Gibson-

Reinemer et al. 2017), and hoop nets to capture juvenile and adult bigheaded carp. We used a MBS-2D Wisconsin control box (ETS Electrofishing LLC, Madison, Wisconsin) powered by a 3,500 watt generator mounted on a 16 foot jon boat, and a larger boat equipped with Smith-Root 5.0 GPP box (Smith-Root, Inc. Vancouver, Washington) for electrofisher sampling. We also used two different electrofishing methods: High

Frequency using pulsed DC current at 60 Hz with a target amperage of 20 amps (jon boat) or 10 amps (large boat), and Low Frequency using pulsed DC current at 15 Hz with a target amperage of four amps. Voltage output was adjusted on both boats to account for changes in water temperature and conductivity to reach amperage goals. Electrofishing runs for the High Frequency method were conducted from downstream to upstream in a manner similar to the method described in Bouska et al. (2017). Runs for the Low

Frequency method were conducted from upstream to downstream. Both methods had run lengths of approximately 30 minutes, focusing on channel sides, backwater habitats, and 25 side channels (DeGrandchamp et al. 2008). Each sampling site was sampled once a month during the summer when water conditions allowed. Run locations were chosen randomly within each sampling site.

We also used an electrified dozer trawl (Hammen et al. 2019) for three days in

2018 and 2019. The electrified dozer trawl (Dozer Trawl) runs were conducted from downstream to upstream at 30 Hz with a target amperage of 30 amps, adjusting voltage to meet amperage goals, and had run lengths of five minutes. Each run location was chosen at random every rkm and varied between the north bank, south bank, and thalweg. Mini- fyke nets (two, 121.9 x 61 cm box frames with a 4 m lead, two, 64.8 cm diameter hoops, and 7 mm mesh netting) were deployed as described in Hubert (1983) in shallow (<1 m) backwater areas and fished overnight. Hoop nets (7 hoops, 91.44 cm in diameter with two throats and 2.5 cm mesh) were deployed as described in Hubert (1983) along the main channel.

All fishes captured were identified to species, measured in total length (TL:mm), and weighed (nearest gram). Sex of all bigheaded carp were identified. Unidentifiable specimens were preserved in 10% formalin and identified in the lab. Lapilli otolith were extracted from bigheaded carp for aging analysis (Seibert and Phelps 2013; Hayer et al.

2014).

Laboratory Methods

We used deionized (DI) water to clean excess tissue off of the otoliths, then mounted the otoliths in epoxy (Epoxicure Epoxy Resin and Hardener, Beuhler Inc., Lake

Bluffs, IL) and sectioned the otoliths across the transverse plane through the nucleus 26 using an ISOMET (Model: 11-1280-160, Beuhler Inc., Lake Bluffs, IL) low-speed saw.

Sections were then sanded using 1,500 and 3,000 grit sandpaper and polished using 3µm lapping paper to reveal annuli. Ages were estimated by examining the polished section beneath a dissecting scope and counting the annuli. Two independent readers estimated the ages of each fish, and discrepancies were reconciled by a consensus between the readers (Stuck et al. 2015). Back-calculated length-at-age measurements were obtained by measuring each annuli using the ImageJ tool in the Fiji software package (Schindelin et al. 2012).

Data Analysis

All data were tested for normality with Shapiro-Wilks tests and visually inspected with density plots. Data that were not normally distributed were analyzed using non- parametric tests. Relative abundance was estimated by calculating catch per unit of effort

(CPUE) in number of bigheaded carp captured per hour of electrofishing. Catch per unit effort data were assessed with Kruskal-Wallis tests, and combined across years if there was no difference in catch rates in each segment between sampling years. Combined data was then tested for differences between segments and among gears for the Dozer Trawl,

High Frequency, and Low Frequency electrofishing methods with Kruskal-Wallis tests followed by a Dunn’s test for pair-wise comparisons. The P-value was adjusted using the

Holmes method after the Dunn’s test. Differences in length distribution between capture methods were assessed by grouping bigheaded carp by method of capture and placing them in 25 mm length bins, and comparing length frequency distributions with 27 bootstrapped Kilmogorov-Smirnov (KS) cumulative distributions tests. The P-value was adjusted for the pairwise KS tests using the Bonferroni method.

We analyzed condition using Analysis of Covariance (ANCOVA) to test differences in log transformed length-weight regression lines of adult bigheaded carp

(>400 mm) across sex, sampling years, and segment of capture, respectively in a hierarchical fashion. If differences were detected in any covariates then the subsequent models were nested inside the proceeding covariate. For example, if differences in the slope of the regressions were detected across sampling years, then models assessing the slope of the regressions across segments would be nested within each year.

Differences in size structure were assessed by grouping fish by segment of capture and placing them in 25 mm length bins. Differences in the length-frequency distributions among river segments were tested using a bootstrapped KS cumulative distributions test. Lengths of sexually mature bigheaded carp (>400 mm) from each segment were compared using an ANOVA. Likewise, we compared ages of adults between segments with a Kruskal-Wallis test. We also assessed year class strength with all aged carp. We used back calculated length-at-age to assess age and growth using the

Dahl-Lea method (Dahl 1907; Lea 1910) for each segment and tested for differences between segments with an ANCOVA (Isely and Grabowski 2007). Growth was then modeled for each segment using Von-Bertalanffy growth functions:

-K(t-t0) Lt = L∞(1-e ) where Lt is the mean length-at-age t, L∞ is the maximum mean length, K relates how quick the function approaches L∞, and t0 is the length at age 0 (Ogle 2016). 28

Results

A total of 1,612 Silver Carp (Table 2-3) and 39 Bighead Carp (Table 2-4) were captured from Segments 1 and 2 over the course of this study. No Silver Carp or Bighead

Carp were captured upstream of Bowersock Dam in Segment 3 (Table 2-1), and too few

Bighead Carp were captured to examine differences in population demographics between segments. Therefore, the subsequent analysis focused on population demographics of

Silver Carp in Segments 1 and 2. Hoop nets failed to capture any species of bigheaded carp and had multiple gear malfunctions due to silting in of the net, and thus were not included in this analysis. Low water conditions in 2018 limited Dozer Trawl sampling largely to Segment 1, but in 2019 effort was equally divided among Segments 1, 2, and 3.

There was no difference in CPUE between High Frequency and Low Frequency electrofishing methods (χ2 =0.372, P = 0.634) in Segment 1. However, the Dozer Trawl catch rates were greater than both the High Frequency (χ2 = 12.82, P < 0.001) and Low

Frequency (χ2 = 20.24, P < 0.001) methods (Figure 2-2) in Segment 1. The High

Frequency electrofishing method and the Dozer Trawl had a higher CPUE than Low

Frequency method (χ2 = 29.32, P = < 0.001; χ2 = 3.3245, P = 0.0432) in Segment 2

(Figure 2-5). Length-frequency distributions were different between Low Frequency and

High Frequency methods (n boots = 5000, D = 0.103, P = 0.008), Dozer Trawl and Low

Frequency methods (n boots = 5000, D = 0.486, P < 0.001), and Dozer Trawl and High

Frequency methods (n boots = 5000, D = 0.551, P < 0.001). The Dozer Trawl captured more juvenile Silver Carp (<400 mm) than both the High Frequency and Low Frequency 29 methods combined, whereas the High Frequency and Low Frequency methods had high catch frequencies of adult carp (Figure 2-7).

The CPUE of Silver Carp was different between Segments 1 and 2 for both the

High Frequency (χ2 = 4.47; P = 0.035) and Low Frequency (χ2 = 42.36; P < 0.001) electrofishing methods. Catch per unit effort was higher in Segment 1with both methods than Segment 2 (Figure 2-2). The mean length of adult Silver Carp captured in Segment 1 was different than in Segment 2 (F1; 1,152 = 410.47, P < 0.001), with Silver Carp in

Segment 2 having longer mean total lengths (Table 2-2). Additionally, the length- frequency distributions were different between segments (n boots = 5000, D = 0.583, P <

0.001). Multiple year-classes were observed in Segment 1, with the 2018 year class dominating the catch of juvenile carp in both 2018 and 2019. Juvenile carp were observed in Segment 2 only in 2019 (Figure 2-3).

There were no differences in the slopes of the length-weight regressions between male and female Silver Carp (F1:793 = 0.907, P = 0.404). There was a difference in the y- intercepts between sampling years (F1:1,127 = 646.6, P < 0.001). The length-weight regression was different between years in both Segment 1 (F1:668 = 16.82, P < 0.001) and

Segment 2 (F1:454 = 6.753, P = 0.01). There were also differences in the length-weight relationships between segments in both 2018 (F1:337 = 6.221, P = 0.013) and 2019 (F1:786

= 8.777, P = 0.003) (Figure 2-4).

Ages were estimated for 73 Silver Carp in Segment 1 and 114 Silver Carp in

Segment 2. The distribution of ages of adult Silver Carp (>400 mm) were not different between Segments 1 and 2 (χ2 = 10.49, P = 0.1056; Table 2). Number of fish recruited 30 was similar between Segments 1 and 2, except in 2014 and 2018. Segment 2 had a larger peak in 2014 than Segment 1; conversely, Segment 1 had a greater peak in 2018 than

Segment 2. No recruits from the 2017 year class were documented (Figure 2-9). Back- calculated length-at-age was performed for 31 Silver Carp in Segment 1 and 60 Silver

Carp in Segment 2. Individual Silver Carp were not all aged in the same direction from the nucleus; therefore, back calculations were performed for fewer individuals to keep the direction consistent. Growth was different between segments (F1:583 = 21.69, P < 0.001) with Silver Carp in Segment 2 exhibiting faster growth rates than Silver Carp in Segment

1 (Figure 2-5). In both segments, Silver Carp grew quickly reaching approximately 500 mm TL in Segment 2 and 450 mm TL in Segment 1 after their second growing season.

Growth leveled off after age four, or about 570 mm TL in Segment 2 and 500 mm TL in

Segment 1 (Figure 2-5).

Discussion

Traditional electrofishing approaches indicate Silver Carp of all sizes were not effectively sampled because of the lack of juveniles captured with these gears (Figure 2-

7). Substantial effort has been spent attempting to remedy this problem (e.g., Bouska et al. 2017; Hammen et al. 2019). Our results indicate the Dozer Trawl captured a wide range of sizes compared to conventional electrofishers. The combination of the push trawl and electrofishing configuration that comprise a Dozer Trawl likely limited size biases associated with conventional electrofishing methods such as netter error (e.g.,

Reynolds 1983; Chick et al. 1999; Bayley and Austen 2002). The push trawl aspect of this gear is particularly advantageous in turbid systems where visibility is an issue. 31

Additionally, the Dozer Trawl is more efficient than conventional electrofishers when this gear can be implemented in ideal conditions (i.e. slow moving habitats with consistent depths > 1m). Variable water levels, flows, and habitat conditions in Segments

2 and 3 limited where the Dozer Trawl could be fully deployed. The Dozer Trawl requires less man power and running time (Hammen et al. 2019), covering longer reaches in shorter periods of time while also reducing costs of monitoring protocols.

Incorporating the Dozer Trawl into our sampling design facilitated the capture of Silver

Carp at rates likely more indicative of their true abundance. Higher catch rates and wider size ranges of Silver Carp captured indicate the Dozer Trawl would be the ideal gear for removal efforts in the Kansas River if this gear were available. However, constructing and maintaining this specialized gear could prove to be too costly for management agencies.

When specialized gears like the Dozer Trawl are not available for use, managers should consider different methods of conventional electrofishing. In reaches with high densities of Silver Carp, we found no difference in CPUE between the High Frequency and Low Frequency methods. At low densities, the High Frequency method was more efficient than the Low Frequency method, which was similar to findings in Bouska et al.

(2017). Both methods of conventional electrofishing were biased toward larger bodied fish (Figure 2-7) and incorporating a suite of gears – such as mini-fyke nets (Collins et al.

2017; Gibson-Reinemer et al. 2017) – may be necessary to sample a broader range of fish sizes. Mini-fyke nets deployed in Segment 1 captured fewer young of year (Table 2-3) than those deployed in other systems, such as the Mississippi River (Haupt and Phelps 32

2016), likely because of the lack of ideal habitat for gear deployment (i.e., backwater or side channel habitats <1m deep). However, the use of mini-fyke nets in the Kansas River did provide utility in monitoring for juvenile carp and locating areas and habitat where juvenile carp occurred.

We failed to detect the presence of Silver Carp upstream of Bowersock Dam over the course of this study (Table 2-3). Bowersock Dam is a known barrier for other fishes such as Blue Sucker Cycletpus elongates (Eitzmann et al. 2007) and Blue Catfish

Ictalurus furcatus (Dean 2020) and appears to be a barrier to longitudinal movements of

Silver Carp as well. Similarly, the Johnson County Weir may serve as at least a partial barrier for Silver Carp as relative abundance of Silver Carp was lower in Segment 2 than

Segment 1, and juvenile carp were scarce in Segment 2 but abundant in Segment 1. A rise in river stage cues movement of adult Silver Carp (DeGrandchamp et al. 2008) upstream in the spring (Coulter et al. 2016) and during times when river conditions are conducive for fish to traverse this barrier, facilitating immigration between Segments 1 and 2. This barrier drastically limits or completely blocks upstream passage during low water periods. Increased movement in response to a rise in river flood stage has been documented in adult Silver Carp (DeGrandchamp et al. 2008; Coulter et al. 2016), but it is unknown if the same response is elicited from juveniles. Juveniles have reduced swimming and burst swimming speeds (Hoover et al. 2012) that may limit river velocities they are able to navigate. Higher flows required to traverse this barrier could prove difficult for juvenile carp to navigate, creating a water velocity barrier. Therefore, we 33 hypothesize that passage of juvenile Silver Carp over the Johnson County Weir may be limited compared to their adult counterparts.

Immigration between Segments 1 and 2 has been documented for Blue Catfish

(Dean 2020) and Shovelnose Sturgeon Scaphirhynchus platorynchus (Werner, unpublished data). Further modifications on the Johnson County Weir aimed at limiting

Silver Carp movement would likely impede movement of other riverine fishes.

Continuity between segments, even if it is limited, may be important for certain life- history traits of native species; therefore, other control measures for bigheaded carp should be considered.

Silver Carp in Segment 2 occurred in lower abundance, had quicker growth rates, and attained longer total lengths than fish from Segment 1. These differences share similar characteristic of reported discrepancies between populations in predominantly free flowing reaches (i.e., Wabash River) and populations in heavily modified reaches

(i.e., Illinois River) (Stuck et al. 2015). Variations in habitat between Segments 1 and 2 follow this pattern in population characteristics. Segment 1 has a higher proportion of urban influence and rip-rap banks, fewer islands and channels, a narrower bankfull width

(Paukert and Makinster 2009), and consistent depths (Eitzmann and Paukert 2010).

Segment 2 is characterized by more variable depths, numerous sandbars, and a further agricultural riparian influence (Paukert and Makinster 2009). Lower abundances in predominantly free flowing reaches may reduce intraspecific competition (Stuck et al.

2015) allowing for greater somatic growth (Amundsen et al. 2007). Additionally, systems with fewer modifications have more habitat and resource availability for native species 34

(Rolls et al. 2012), maintaining functional redundancy (Rosenfeld 2002) and limiting resources available to invading fishes. Therefore, population demographics of Silver Carp in Segment 2, coupled with less anthropogenic alterations, support the hypothesis that systems with fewer modifications are more resilient to large-scale invasions of Silver

Carp (Stuck et al. 2015).

Decreased rates of intraspecific competition for Silver Carp in Segment 2 could also be influencing discrepancies in condition observed in 2018 (Figure 2-4). Food consumption rates have been shown to exhibit a curvilinear relationship with population density. Populations with greater abundances have reduced consumption rates resulting in poorer condition and reduced growth rates (Amundsen et al. 2007). Differences in body condition between Silver Carp populations in Segments 1 and 2 may have been amplified by lower flows in 2018. There was little variation in condition between segments in 2019; however, Silver Carp had lower weights at a given length in 2019 than in 2018. River discharge was drastically different between sampling years (Figure 2-8), where discharge in 2018 was consistently at 42 m2 second-1 during the sampling season. Discharge was sporadic in 2019 with multiple high water events; the largest of which was approximately

2,800 m3 second-1. High flows could have been an additive stressor on these fish, requiring a greater metabolic demand to navigate increased river velocities. Additionally, turbidity in the Kansas River was higher in 2019 than in 2018 (Figure 2-8). Elevated turbidity and flows in large rivers can lead to decreases in gross primary production by decreasing light transmittance and respiration. High velocity flows inhibit the buildup of autotroph biomass and reduces residence time (Glibert et al. 2014; Bernhardt et al. 2018; 35

Hosen et al. 2019). Reduced food availability coupled with higher metabolic demand likely contributed to the reduced condition of Silver Carp in 2019. We did not take measurements to assess gonadal somatic index (GSI) in this study, but we did observe fewer gravid females in 2019. However, lack of gonadal development in females in 2019 likely had negligible influence on measures of condition because we found no effect of sex on length-weight relationships.

The scarcity of juvenile Silver Carp in Segment 2 indicates reproduction may be limited in this segment of the river. Additionally, young-of-year Silver Carp were more abundant during the low water year in 2018 (Figure 2-3). Presence of larval Silver Carp in this system has never been investigated, but microchemistry analysis indicate production occurring in the Kansas River is the greatest source for Silver Carp in this system (Chapter 4). Additionally, young-of-year Silver Carp occur at higher densities in areas with slower moving waters (e.g., Haupt and Phelps 2016) and could be using tributaries of the Missouri River as nursery habitats, seeking refuge from the high velocity currents of the Missouri River.

Tributaries to the Missouri River system – such as the Kansas River – may be important for completing life-history stages, particularly for life stages that require conditions not typically found within the main-stem of the Missouri River. For example,

Segment 1 of the Kansas River could be a source for Silver Carp production in the

Missouri River, as well as upstream segments of the Kansas River (Chapter 4). Lack of recruitment in Segment 2 indicate that this segment may be functioning as a sink for

Silver Carp immigrating from Segment 1. Focusing removal efforts to Segment 1 could 36 have trickle-down effects to Segment 2 as well as the Missouri River. Removal efforts may be particularly effective during years with abundant YOY Silver Carp that are confined to Segment 1. Continued monitoring of Segment 3 is needed to detect early invasion and for subsequent eradication attempts before Silver Carp are able to become established. Investigating how Silver Carp function in the multitude of tributaries to the

Missouri River will provide a greater holistic understanding of how these carp are functioning throughout the drainage. Such information is invaluable when formulating and implementing a basin wide control plan. 37

Literature Cited Allen, M. S., and J. E. Hightower. 2010. Fish population dynamics: mortality, growth and recruitment. Pages 43-80 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Anderson, R. O. and S. J. Gutreuter. 1983. Length, weight, and associated structural indices. Pages 283-300 in L.A Nielsen and D.L. Johnson, editors. Fisheries Techniques. American Fisheries Society, Bethesda, Maryland. Amundsen, P-A., R. Knudsen, and A. Klemetsen. 2007. Interspecific competition and density dependence of food consumption and growth of Arctic char. Journal of Animal Ecology 76:149-158. Bayley, P. B. and D. J Austen. 2002. Capture efficacy of a boat electrofisher. North American Journal of Fisheries Management 131:435-451. Bernhardt, E. S., J. B. Heffernan, M. B. Grimm, E. H. Stanley, J. W. Harvey, M. Arroita, A. P. Appling, M. J. Cohen, W. H. McDowell, R. O. Hall, Jr., J. S. Read, B. J. Roberts, E. G. Stets, and C. B. Yackulic. 2018. The metabolic regimes of flowing rivers. Limnology and Oceanography 63:S99-S118 Bottcher, J. L., P. Budy, and D. S. Speas. 2013. Frequent usage of tributaries by the endangered fishes of the Upper Colorado River Basin; Observations from the San Rafael River, Utah. North American Journal of Fisheries Management 33:585- 594. Bouska, W. W., D. C. Glover, K. L. Bouska, and J. E. Garvey. 2017. A refined electrofishing technique for collecting Silver Carp: implications for management. North American Journal of Fisheries Management 37:101-107. Brönmark, C., K. Hulthén, P. A. Nilsson, C. Skov, L.A. Hansson, J. Brodersen, and B. B. Chapman. 2014. There and back again: migration in freshwater fishes. Canadian Journal of Zoology 92:467-479. Calkins, H. A., S. J. Tripp, and J. E. Garvey. 2012. Linking silver carp habitat selection to flow and phytoplankton in the Mississippi River. Biological Invasions 14:949- 958. Chaffin, B. C., A.S. Garmestani, D. G. Angeler, D. L. Herrmann, C. A. Stow, M. Nyström, J. Sendzimir, M. E. Hopton, J. Kolasa, and C. R. Allen. 2016. Biological invasions, ecological resilience and adaptive governance. Journal of Environmental Management 183:399-407 Chick, J. H., S. Coyne, and J.C. Trexler. 1999. Effectiveness of airboat electrofishing for sampling fishes in shallow, vegetated habitats. North American Journal of Fisheries Management 19:957-967 38

Chick, J. H., D. K. Gibson-Reinemer, L. Soeken-Gittinger, A. F. Casper. 2020. Invasive Silver Carp is empirically linked to declines of native sportfish in the Upper Mississippi River system. Biological Invasions 22:723-734.012 Collins, S. F., M. J. Diana, S. E. Butler, and D. H. Wahl. 2017. A comparison of sampling gears for juvenile silver carp in river-floodplain ecosystems. North American Journal of Fisheries Management 37:94-100. Conover, G., R. Simmons, and M. Whalen. 2007. Management and Control plan for Bighead, Black, Grass, and Silver Carps in the United States. Asian carp Working Group, Aquatic Nuisance Species Task Force, Washington D.C. 223 pp. Coulter, A. A., E. J. Bailey, D. Keller, and R. R. Goforth. 2016. Invasive Silver Carp movement patterns in the predominantly free-flowing Wabash River (Indiana, USA). Biological Invasions 18:471-485. Dahl, K. 1907. The scales of the herring as a means of determining age, growth and migration. Report on Norwegian Fishery and Marine-Investigations 2:1-39. Dean, Q. J. 2020. Population characteristics and movement of Blue Catfish in the Kansas River. Master’s thesis. University of Nebraska, Lincoln. DeBoer, J. A., A. M. Anderson, and A. F. Casper. 2018. Multi-trophic response to invasive Silver Carp (Hypophthalmichthys molitrix) in a large floodplain river. Freshwater Biology 63:597-611. DeGrandchamp, K. L., J. E. Garvey, and R. E. Colombo. 2008. Movement and habitat selection by invasive Asian carp in a large river. Transactions of the American Fisheries Society 137:45-56. DeGrandchamp, K. L., J. E. Garvey, and L. A. Csoboth. 2007. Linking adult reproduction and larval density of invasive carp in8 a large river. Transactions of the American Fisheries Society 136:1327-1334. Deters, J. E., D. C. Chapman, and B. McElroy. 2013. Location and timing of Asian carp spawning in the lower Missouri river. Environmental Biology of Fishes 96:617- 629. Eitzmann, J. L., A. S. Makinster, and C. P. Paukert. 2007. Distribution and growth of blue sucker in a great plains river, USA. Fisheries Management and Ecology 14:255-262. Eitzmann, J. L. and C. P. Paukert. 2010. Longitudinal differences in habitat complexity and fish assemblage structure of a great plains river. The American Midland Naturalist 163:14-32. Freeze, M. and S. Henderson. 1983. Distribution and Status of the Bighead Carp and Silver Carp in Arkansas. North American Journal of Fisheries Management 2:197-200. 39

Galat, D. L., L. H. Fredrickson, D. D. Humburg, K. J. Bataille, J. R. Bodie, J. Dohrenwend, G. T. Gelwicks, J. E. Havel, D. L. Helmers, J. B. Hooker, J. R. Jones, M. F. Knowlton, J. Kubisiak, J. Mazourek, A. C. McColpin, R. B. Renken, and R. D. Semlitsch. 1998. Flooding to restore connectivity of regulated, large- river wetlands. BioSciene 48:721-733. Gibson-Reinemer, D. K., L. E. Solomon, R. M. Pendleton, J. H. Chick, and A. F. Casper. 2017. Hydrology controls recruitment of two invasive cyprinids: bighead carp reproduction in a navigable large river. PeerJ 5:e3641. Glibert, P. M., R. C. Dugdale, F. Wilkerson, A. E. Parker, J. Alexander, E. Antell, S. Blaser, A. Johnson, J. Lee, T. Lee, S. Murasko, and S, Strong. 2014. Major – but rare – spring blooms in 2014 in San Francisco Bay Delta, California, a result of the long-term drought, increased residence time, and altered nutrition loads and forms. Journal of Experimental Marine Biology and Ecology 460:8-18. Gozlan, R. E., J. R. Britton, I. Cowx, and G. H. Copp. 2010. Current knowledge on non- native freshwater fish introductions. Journal of Fish Biology 76:751-786. Hamel, M. J., M. A. Pegg, J. J. Hammen, and M. L. Rugg. 2014. Population characteristics of Pallid Sturgeon, Scaphirhynchus albus (Forbes & Richardson, 1905), in the lower Platte River, Nebraska. Journal of Applied Ichthyology 30:1362-1370. Hammen, J. J., E. Pherigo, W. Doyle, J. Finley, K. Drews, and J. M. Goeckler. 2019. A comparison between conventional boat electrofishing and the electrified dozer trawl for capturing Silver Carp in tributaries of the Missouri River, Missouri. North American Journal of Fisheries Management 39:582-588. Haupt, K. J., and Q. E. Phelps. 2016. Mesohabitat association in the Mississippi River Basin: a long-term study on the catch rates and physical habitat association of juvenile Silver Carp and two native planktivores. Aquatic Invasions 11:93-99. Hayer, C.-A., J. J. Breeggemann, R. A. Klumb, B. D. S. Graeb, and K. N. Bertrand. 2014. Population characteristics of Bighead and Silver Carp on the Northwestern front of their North American invasion. Aquatic Invasions 9:289-303. Hoover, J. J., W. Southern, A. W. Katzenmeyer, and N. M. Hahn. 2012. Swimming performance of bighead carp and silver carp: Methodology, metrics, and management applications. ANSRP Technical Notes Collection. ERDC/TN ANSRP-12-3. Vicksburg, MS: U.S. Army Engineer Research and Development Center. Hosen, J. D., K. S. Aho, A. P. Appling, E. C. Creech, J. H. Fair, R. O. Hall Jr. E. D. Kyzivat, R. S. Lowenthal, S. Matt, J. Morrison, J. E. Saiers, J. B. Shanley, L. C. Weber, B. Yoon, and P. A. Raymond. 2019. Enhancement of primary production during drought in a temperate watershed is greater in large rivers than headwater streams. Limnology and Oceanography 64:1458-1472. 40

Hubert, W. A. 1983. Passive capture techniques. Pages 95-122 in L. A. Nielsen and D. L. Johnson, editors. Fisheries Techniques. American Fisheries Society, Bethesda, Maryland. Irons, K. S., G. G. Sass, M. A. McClelland, and J. D. Stafford. 2007. Reduced condition factors for two native fish species coincident with invasion of non-native Asian carps in the Illinois River, U.S.A. Is this evidence for competition and reduced fitness? Journal of Fish Biology 71:258-273. Isely, J. J., and T. B. Grabowski. 2007. Age and Growth. Pages 187-228 in C. S. Guy and M. L. Brown, editors. Analysis and interpretation of freshwater fisheries data. American Fisheries Society, Bethesda, Maryland. Jerde, C. L., W. L Chadderton, A. R. Mahon, M. A. Renshaw, J. Corush, M. L. Budney, S. Mysorekar, and D. M. Lodge. 2013. Detection of Asian carp DNA as part of a Great-Lakes basin wide surveillance program. Canadian Journal of Fisheries and Aquatic Sciences 70:522-526. Kolar, C. S., D. C. Chapman, W. R. Courtenay Jr., C. M. Housel, and J. D. Williams. 2005. Asian carps of the Genus Hypophthalmichthys (Pisces, Cyprinidae) – A biological synopsis and environmental risk assessment. National Invasive Species Council materials. 5. Kolar, C. S., W. R. Courtenay, and L. G. Nico. 2010. Managing undesired and invading fishes. Pages 213-259 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Kolar, C. S., and D. M. Lodge. 2002. Ecological predictions and risk assessment for alien fishes in North America. Science 298:1233-1236. Lea, E. 1910. On the methods used in the herring investigations. Publications de Circonstance, Conseil Permanent International pour l’Exploration de la Mer 53:7- 25. MacNamara, R., D. Glover, J. E. Garvey, W. Bouska, and K. Irons. 2016. Bigheaded carps (Hpophthalmichthys spp.) at the edge of their invaded range: using hydroacoustics to assess population parameters and the efficacy of harvest as a control strategy in a large North American river. Biological Invasions 18:3293- 3307. Makinster, A. S., and C. P. Paukert. 2008. Effects and utility of minimum length limits and mortality caps for Flathead Catfish in discrete reaches of a large prairie river. North American Journal of Fisheries Management 28:97-108. Mosher, T. D. 2014a. Bighead Carp, Hypophthalmichthys nobilis Richardson 1845. Pages 166-168 in Kansas Fishes Committee. Kansas fishes. University Press of Kansas, Lawrence. 41

Mosher, T .D. 2014b. Silver Carp, Hypophthalmichthys molitrix Richardson 1845. Pages 166-167 in Kansas Fishes Committee. Kansas gishes. University Press of Kansas, Lawrence. Neely, B. C., M. A. Pegg, and G. E. Mestl. 2009. Seasonal use distributions and migrations of blue sucker in the Middle Missouri River. Ecology of Freshwater Fishes 18:437-444. Norman, J. D., and G. W. Whitledge. 2015. Recruitment sources of invasive bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) inhabiting the Illinois river. Biological Invasions 17:2999-3014. Ogle, D. K. 2016. Introductory fisheries analysis with R. CRC Press, New York, NY pp. 221-250. Paukert, C. P., and A. S. Makinster. 2009. Longitudinal patterns in Flathead Catfish relative abundance and length at age within a large river: effects of an urban gradient. River Research and Applications 25:861-873. Pegg, M. A., C. L. Pierce, and A. Roy. 2003. Hydrological alteration along the Missouri River Basin: A time series approach. Aquatic Sciences 65:63-72. Quist, M. C., and C. S. Guy. 1999. Spatial variation in population characteristics of the Shovelnose Sturgeon in the Kansas River. The Prairie Naturalist 31:65-74. Quist, M. C., J. S. Tillma, M. N. Burlingame, and C. S. Guy. 1999. Overwinter habitat use of shovelnose sturgeon in the Kansas River. Transactions of the American Fisheries Society 128:522-527. Reynolds, J. B. 1983. Electrofishing. Pages 147-164 in L. A. Nielsen and D. L. Johnson, editors. Fisheries Techniques. American Fisheries Society, Bethesda, Maryland. Rolls, J. R., C. Leigh, and F. Sheldon. 2012. Mechanistic effects of low-flow hydrology on riverine ecosystems: ecological principles and consequences of alteration. Freshwater Science 31:1163-1186. Rosenfeld, S. J. 2002. Functional redundancy in ecology and conservation. Oikos 98:1. Sampson, S. J., J. H. Chick, and M. A. Pegg. 2009. Diet overlap among two Asian carp and three native fishes in backwater lakes on the Illinois and Mississippi rivers. Biological Invasions 11:483-496. Sass, G. S., C. Hinz, A. C. Erickson, N. N. McClelland, M. A. McClelland, and J. M. Epifanio. 2014. Invasive bighead and silver carp effects on zooplankton communities in the Illinois River, Illinois USA. Journal of Great Lakes Research 40:911-921.

Schindelin, J., I. Arganda-Carreras, and E. Frise. 2012. Fiji: an open-source platform for biological image analysis. Nature Methods 9:676-682. 42

Seibert, J. R., and Q. E. Phelps. 2013. Evaluation of aging structures for silver carp from Midwestern U.S. rivers. North American Journal of Fisheries Management 33:839-844.

Solomon, L. E., R. M. Pendleton, J. H. Chick, and A. F. Casper. 2016. Long-term changes in fish community structure in relation to the establishment of Asian carp in a large floodplain river. Biological Invasions 18:2883-2895.

Steffensen, K. D. and G. E. Mestl. 2016. Assessment of Pallid Sturgeon relative condition in the upper channelized Missouri River. Journal of Freshwater Ecology 31:583- 595.

Stuck, J. G., A. P. Porreca, D. H. Wahl, and R. E. Colombo. 2015. Contrasting population demographics of invasive silver carp between and impounded and free-flowing river. North American Journal of Fisheries Management 35:114-122.

Wanner, G. A., and R. A. Klumb. 2009. Asian carp in the Missouri river: analysis from multiple Missouri river habitat and fisheries programs. National Invasive Species Council materials. Paper 10.

White, K., J. Gerken, C. P. Paukert, and A. Makinster. 2010. Fish community structure in natural and engineered habitats in the Kansas River. River Research and Applications 26:797-805. Williamson, C. J., and J. E. Garvey. 2005. Growth, fecundity, and diets of newly established silver carp in the middle Mississippi river. Transactions of the American Fisheries Society 134:1423-1430. 43

Table 2-1: Median, first, and third quartiles of catch per unit effort (number/hour) of

Silver Carp with the High Frequency electrofishing (HF), Low Frequency electrofishing

(LF), and the Dozer Trawl in Segments 1, 2, and 3 of the Kansas River.

Segment Gear 1st Quartile Median 3rd Quartile LF 3.31 9.83 18.95 1 HF 2 17.42 25.71 Dozer Trawl 15 102 162 LF 0 0 4 2 HF 2 6.33 13.78 Dozer Trawl 0 12 12 LF 0 0 0 3 HF 0 0 0 Dozer Trawl 0 0 0

44

Table 2-2: Mean lengths and ages of adult Silver Carp (>400 mm) captured in Segments

1 and 2 of the Kansas River. Standard deviation provided in parenthesis. Age estimated from lapilli otolith sections. Segment 1 Segment 2

Total Length 592.7 (49.3) 657.3 (58.6)

Age 5.6 (1.3) 5.2 (1.1)

45

Table 2-3: Total catch of all Silver Carp per segment in 2018 and 2019 from the Kansas

River with all gears deployed.

Silver Carp

Segment Gear Type 1 2 3 Total Catches High Frequency EF 215 280 0 495 Low Frequency EF 563 152 0 715 Dozer Trawl 348 39 0 387 Mini-fyke Nets 13 2 0 15 Total 1,139 473 0 1,612

46

Table 2-4: Total catch of all Bighead Carp per segment in 2018 and 2019 from the

Kansas River with all gears deployed.

Bighead Carp

Segment Gear Type 1 2 3 Total Catches High Frequency EF 8 6 0 14 Low Frequency EF 11 12 0 23 Dozer Trawl 1 0 0 1 Mini-fyke Nets 1 0 0 1 Total 21 18 0 39

47

Figure 2-1: Kansas River study area where Segment 1 (24 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (61 rkm) is between the Johnson County Weir and Bowersock Dam, and Segment 3 (57 rkm) is between Bowersock Dam and the Topeka Weir.

48

Figure 2-2: Mean and 95% confidence intervals of catch per unit effort (CPUE; number/hour) for the Dozer Trawl, High Frequency electrofishing, and Low Frequency electrofishing methods for Segments 1 and 2 of the Kansas River. Error bars represent the

95% confidence intervals for the mean.

49

Figure 2-3: Length-frequency histograms of Silver Carp captured in Segments 1 and 2 of the Kansas River. Silver Carp were separated into 25 mm length bins and enumerated.

Light grey bars represent Silver Carp captured in 2018 and dark grey bars represent

Silver Carp captured in 2019; 2018 bars are stacked on top of 2019 bars.

50

Figure 2-4: Regressions of log10 transformed lengths and weights of adult Silver Carp

(>400 mm) captured from Segments 1 (light gray) and 2 (dark gray) in 2018 (solid lines) and 2019 (dashed lines) in the Kansas River.

51

-0.2159(t-(-0.802) Segment 1: Lt = 796.9(1-e )

-0.3.433(t-(-0.16) Segment 2: Lt = 744.1(1-e )

Figure 2-5: Length-at-age of Silver Carp in Segments 1 (light gray circles) and 2 (dark gray triangles); data points at each age are offset for clarity. Von-Bertalanffy growth curves were modeled for Segment 1 (light gray line) and Segment 2 (dark gray line).

Growth curves were not offset.

52

Figure 2-6: Predicted lengths-at-age from Von-Bertalanffy growth models for Silver

Carp populations in the Illinois River from Grafton, IL to Ottowa IL (red circles and solid line), Segment 1 of the Kansas River (light gray triangles and dotted line), Segment 2 of the Kansas River (dark gray squares and dashed line), and Wabash River from New

Haven, IL to Darwin, IL (blue crosses and dashed line) (Stuck et al. 2015).

53

Figure 2-7: Length frequency histograms of Silver Carp captured with Dozer Trawl,

High Frequency electrofishing, and Low Frequency electrofishing from the Kansas River in both 2018 and 2019. Silver Carp were separated into 25 mm length bins and enumerated.

54

Figure 2-8: Discharge (blue line), measured in cubic meters per second, and turbidity

(brown line), measured in Formazin Nephelometric Units, in the Kansas River from April of 2018 to October of 2019. Shaded regions represent river conditions during the 2018 sampling season (left) and the 2019 sampling season (right).

55

Figure 2-9: Recruitment of Silver Carp captured from Segments 1 (light grey points and solid line) and 2 (dark grey points and dotted line) of the Kansas River. The number of fish in each year class were determined by back-calculating all specimens whose age was determined from sectioned lapilli otoliths in each year of sampling. 56

CHAPTER 3: EFFICACY OF ENVIRONMENTAL DNA FOR ASSESSING THE DISTRIBUTION OF INVASIVE ASIAN CARP IN A LARGE TURBID RIVER

J. P. WERNER*, Q. J. DEAN, S. FERNANDO, M. A. PEGG and M. J. HAMEL

Abstract

Silver Carp Hypophthalmichthys molotrix, Bighead Carp H. nobilis, and Black

Carp Molypharyngodon piceus, collectively referred to as Asian carp, are a group of invasive fishes continuing to expand their range throughout the lower Missouri River basin. Accurate information on the current distribution of these invasive fishes is crucial to identify waters that are vulnerable to secondary introductions. Additionally, early detection of Asian carp in uninvaded reaches is paramount to developing management plans that prevent carp establishment. Environmental DNA (eDNA) is an emerging tool that can detect rare fishes or fishes in low abundance and has been used for early detection of invasive species. Here, we attempted to use eDNA to monitor Asian carp distribution throughout the lower Kansas River, Kansas, and to assess the ability of a low-head dam to impede upstream infestations. We did not detect any species of Asian carp upstream of the dam, but we did detect Black Carp near the confluence with the

Missouri River. However, our approach failed to detect Silver Carp and Bighead Carp in reaches where these fishes have been documented with conventional gears. These false negative results may have been caused by several factors, including low filtration capacity from the turbid conditions of the Kansas River and low sample volume relative to the volume of water in the river. Many sources of error exist with this analytical tool 57 and the limitations are not fully understood. As such, future work aimed at comparing eDNA use in varying water conditions is warranted.

Introduction

Invasive species are a major threat to native freshwater fishes in North America

(Kolar et al. 2010). Silver Carp Hypophthalmichthys molotrix, Bighead Carp H. nobilis, and Black Carp Molypharyngodon piceus, collectively known as Asian carp, are one group of invasive fishes in the United States (Conover et al. 2007). Asian carp can have large ecological (Irons et al. 2007; Sampson et al. 2009; Sass et al. 2014, Chick et al.

2020) and economic (Conover et al. 2007) impacts in ecosystems in which they are able to invade and become established in. Since their introduction in the 1970’s (Freeze and

Henderson 1982), Asian carp have become widespread throughout the Mississippi River drainage (Conover et al. 2007). Bighead Carp and Silver Carp have been documented throughout the Mississippi and Missouri rivers (Conover et al. 2007) and have expanded their range to as far north as North Dakota (Hayer et al. 2014). Black Carp specimens have been documented in the middle and lower Mississippi River, and in the lower

Missouri River as far upstream as Jefferson City, Missouri (Nico and Nielson 2018).

However, Black Carp distribution is likely greater than current data suggest because they are benthic oriented and difficult to detect (Nico and Jelks 2011).

Silver Carp and Bighead Carp can exhibit extensive longitudinal movements in large river systems (Peters et al. 2006; DeGrandchamp et al. 2008; Coulter et al. 2016).

Such longitudinal movements are often impeded by large natural or anthropogenic 58 barriers (e.g., Porto et al. 1999; Eitzmann et al. 2007; Paukert and Galat 2010; Sparks et al. 2011, Dean 2020). These barriers are often on the edge of Asian carp distributions preventing upstream movement (i.e., Gavin’s Point Dam, SD and CAWS Electric Fish

Dispersal Barrier, IL). However, Asian carp have been able to traverse or circumnavigate some of these barriers (e.g., Moy et al. 2011; Jerde et al. 2013; Tripp et al. 2014) and expand into new waters, also known as secondary introductions (Vander Zanden and

Olden 2008).

Mitigating secondary introductions is crucial to ensuring the resiliency of an ecosystem (Kolar and Lodge 2002; Vander Zanden and Olden 2008). Information on the current distribution of Asian carp is vital to identify vulnerable waters and prevent further secondary introductions. Monitoring waters at a high risk of secondary introductions is needed to quickly implement management strategies that reduce the risk of establishment

(Vander Zanden and Older 2008; Kolar et al. 2010). However, Asian carp are difficult to detect at low densities with conventional gears. Environmental DNA (eDNA) is a powerful tool to detect rare, difficult to detect species, or species in low abundance

(Ficetola et al. 2008; Jerde et al. 2011; Thomson et al. 2012; Mahon et al. 2013; Pilliod et al. 2013; Laramie et al. 2015). For example, this technique has been used to monitor for

Silver Carp and Bighead Carp in water bodies with a high invasion risk such as Lake

Michigan (Jerde et al. 2013).

Silver Carp and Bighead Carp have been documented in the Kansas River,

Kansas, downstream from Bowersock Dam (Figure 3-1). Bowersock Dam is a low-head

(Quist and Guy 1999) hydropower dam that impedes upstream longitudinal movements 59 of other riverine fishes such as Blue Suckers Cycleptus elongatus (Eitzmann et al. 2007) and Blue Catfish Ictalurus furcatus (Dean 2020). However, Bighead Carp were able to traverse this dam during a flood in 1993, but were unable to establish a breeding population upstream of the dam. Silver Carp have never been documented upstream of

Bowersock Dam (C. Steffen, Kansas Department of Wildlife, Parks and Tourism, personal communication). However, multiple high water events in 2019 (Chapter 2) may have facilitated immigration of Silver Carp and Bighead Carp into reaches above

Bowersock Dam. Here, we used eDNA to 1) investigate if Silver Carp and Bighead Carp are present upstream of Bowersock Dam, and 2) assess the presence of Black Carp in the

Kansas River.

Methods

Study Area

The Kansas River originates in North-central Kansas at the confluence of the

Smoky Hill and Republican Rivers (Quist and Guy 1999). The Kansas River then flows easterly 274 river kilometers (rkm) (Makinster and Paukert 2008) to its confluence with the Missouri River near Kansas City, KS. There are three anthropogenic barriers on the main stem of the Kansas River: the Johnson County Weir (rkm 27) in Edwardsville, KS,

Bowersock Dam (rkm 83) in Lawrence, KS, and the Topeka Weir (rkm) 141 in Topeka,

KS. These barriers delineated distinct reaches (i.e., segments) within our study area.

Segment 1 is from the confluence with the Missouri River to the Johnson County Weir

(27 rkm), Segment 2 is between the Johnson County Weir and Bowersock Dam (56 rkm) and Segment 3 is between Bowersock Dam and the Topeka Weir (58 rkm) (Figure 3-1). 60

Sample Collection

We selected 13 sites to collect water samples along the main stem of the Kansas

River. Sites were typically separated by a distance of approximately 10-20 rkm. Sampling sites 1-3 were in Segment 1, sites 4-8 were in Segment 2 and 9-13 were in Segment 3

(Figure 3-1). Six, 1 L surface water samples were collected from each site, two from either bank and two from the middle of the channel. Samples were collected from the bow of a boat using sterilized Whirl-Paks (Whirl-Pak, Madison, WI). Sampling was conducted from downstream to upstream to avoid possible contamination by the boat.

Bank samples were collected approximately two meters from the bank to avoid sediment being stirred up from the wake. Each Whirl Pack was placed in a one gallon Ziplock (S.

C. Johnson & Sons Inc., Racine, WI) bag to help mitigate cross contamination during transport, and then immediately put on ice (Coulter et al. 2019). The hull, bow, and outside of the transportation coolers were sprayed down with a 10% chlorine solution between sites. All samples were frozen at -20ºC within 24 hours of collection until extraction. We collected water samples from August 20-23, 2019. Additional samples (n

= 3) were collected during this time at Site 1 for a pilot study to ensure our extraction and amplification methods were appropriate. Sites 1, 2, 3, 7, and 8 were lost due to a freezer malfunction and were recollected on February, 17, 2020. Site 9 was also recollected on

February, 17, 2020 to control for DNA contamination from anglers using Silver Carp cut bait in Segment 3.

A single Whirl-Pak was filled with distilled water at half the sampling sites as a negative control (i.e., field blank), and was transported in the same manner as the rest of 61 the samples. Positive controls for Silver Carp and Bighead Carp were collected in the field from the live well after a specimen from each species had been held in the tank for at least 30 minutes. The U.S. Fish and Wildlife Service (USFWS) provided a Black Carp positive control. All positive and negative controls were processed using the same methods as the other field samples.

DNA Extraction: Filtration

The Kansas River has elevated amounts of suspended sediment compared to other rivers in a forested ecoregion (Thorp and Mantovani 2005). Elevated sediment loads posed issues when attempting to filter genetic material from water samples because of clogging of filters. Therefore, we were able to extract DNA using filtration methods only from sites 1 – 3 and 7 – 9 from the February, 2020 sampling, as turbidity was much lower during the winter (Table 3-3). Each 1 L sample was slowly thawed at 4ºC and then filtered through a Whatman 934-AHTM 1.5 µm glass fiber filter (GE Whatman,

Fairfield, CT) using a polyphenylsulfone magnetic funnel filter (Pall Corporation, Port

Washington, NY). All filtration equipment was submerged in a 10% chlorine bath for a minimum of 10 minutes. The workspace was wiped down with the 10% chlorine solution between processing each sample (Coulter et al. 2019). Each filter was placed in a sterilized 5ml polyethylene vial and stored at -20ºC. Aqueous DNA was extracted by removing organic material from the surface of the filter using sterilized spatulas. The top layer of filter material was also removed and placed in a 2ml Safe-Lock (Eppendorf AG,

Hamburg, Germany) tube along with the organic material. 62

Aqueous DNA was extracted using a Mag-Bind Stool DNA 96 Kit (Omega Bio- tek, Norcross, GA, USA) according the instructions provided by the manufacturer with an added modification of using a TissueLyser to bead beat our samples (Qiagen Inc.,

Valencia, CA, USA) for 10 minutes at 20Hz prior to incubation in a 90°C water bath for

10 minutes. Precipitation of DNA was also modified. Each sample received 0.2x volume of 3M sodium acetate (pH 5.3) and one volume of 100% isopropanol, and was vortexed.

Samples were then incubated at -20ºC overnight before being centrifuged at 16,000 x g for 15 minutes at 4°C. The supernatant was then discarded and the nucleic acid pellet was washed with 70% ethanol chilled to -20ºC in order to remove residual salt and was centrifuged at 13,000 x g for 2 minutes at room temperature. The ethanol wash was discarded and pellet was dried and dissolved in 300μL of Tris (10mM, pH 8) (Perisho, unpublished data). Further purification of nucleic acids was performed according to the instructions provided by Omega Bio-tek. Extraction of negative controls (i.e., extraction performed on deionized water) were included during each extraction (n = 3) to monitor for contamination.

DNA Extraction: Centrifugation

We extracted DNA from Sites 4 – 6, 9 – 13, and from the pilot study samples from the August 2019 sampling event using centrifugation procedures. Each 1 L sample was slowly thawed at 4°C and divided among four autoclaved 250ml bottles for centrifugation. We centrifuged water samples at 12,700 RPM, resulting in a pellet of organic material. We then preserved each pellet in 600µl of Tris (10mM, pH 8) and placed the pellet in a 2ml centrifuge tube, one centrifuge tube for each 250ml bottle. All 63 organic pellets from each sample were then pooled together and homogenized. We took a

450µl subsample from each sample for extraction, and extracted aqueous DNA using the procedures described above. Extraction of negative controls were included during each extraction to monitor for contamination during extractions (n = 2).

PCR Amplification and Evaluation

We used the primers for Silver Carp and Bighead Carp developed by Jerde et al.

(2011) and Black Carp primers developed by Mahon et al. (2013). Thermal regimes described by Jerde et al. (2011) were modified for Silver Carp and Bighead Carp to reduce streaking in our gels by reducing the number of PCR cycles from 45 to 30.

Additionally, we increased the annealing temperature for Silver Carp from 52ºC to 54ºC.

We used thermal regimes for Black Carp described in Mahon et al. (2013), but decreased the number of cycles from 45 to 40 to reduce streaking. We visualized results using a

1.5% agarose gel stained with ethidium bromide (Jerde et al. 2011; Mahon et al. 2013).

Positive results were classified as a clear band at 191 base pairs (bp) for Silver Carp, 312 bp for Bighead Carp, and 170 bp for Black Carp (Jerde et al. 2011; Mahon et al. 2013).

Asian carp DNA presence was assessed using three polymerase chain reactions (PCR’s) per species for every water sample. If only one of the PCR’s yielded a positive result for a given sample, the PCR process was performed again. The sample would have to yield at least one positive outcome again to be deemed a positive result (Laramie et al. 2015).

Samples were deemed as false negatives when the samples failed to detect Silver Carp or

Bighead Carp in locations where the species are known to occur.

64

Data Analysis

Presence of each species was confirmed by positive eDNA results. The ability of

Bowersock Dam to impede longitudinal upstream movement of Silver Carp and Bighead

Carp was assessed by calculating detection probabilities upstream and downstream of the dam. Detection rates (Dr) were calculated for each species at each sampling site with:

푁푝 퐷 = × 100 푟 푁 where Np is the number of positive detection samples at a site and N is the total number of samples at the site (Laramie et al. 2015). False negative probabilities for Silver Carp and

Bighead Carp were calculated for all samples collected downstream of Bowersock Dam.

False negative rates (Fr) were calculated using:

푆 퐹 = 푛 × 100 푟 푆 where Sn is the number of negative samples downstream of Bowersock Dam and S is the total number of samples collected downstream of the dam.

Results and Discussion

We confirmed that our DNA extraction techniques yielded genetic material by visual confirmation of banding in agarose gels ran before PCR procedures. Additionally, we were able to amplify Asian carp DNA using the described primers from our positive control samples. During the pilot study, we detected Silver Carp (Dr = 100%, n = 3) and

Black Carp (Dr = 100%, n = 3) but failed to detect Bighead Carp (Dr = 0%, n = 3) near the Kansas-Missouri River confluence. We did not detect any of our target species at any other sampling site with either the centrifugation or the filtration procedures (Table 3-1). 65

We observed false negative results from centrifuged samples for Silver Carp (Fr = 86%, n

= 18) and Bighead Carp (Fr = 100%, n = 21). Likewise, we observed false negative results in the filtered samples for both Silver Carp (Fr = 100%, n = 36) and Bighead Carp

(Fr = 100%, n = 36) (Table 3-2).

Our extraction and amplification procedures were successful in isolating DNA, but were unable to detect target DNA for Silver Carp and Bighead Carp in any samples other than the pilot study. The positive detections for Black Carp from the pilot study near the confluence with the Missouri River mark the northernmost detection of Black

Carp within the Missouri River basin. Furthermore, we did not detect any species of

Asian carp above Bowersock Dam in Segment 3 with our eDNA assay (Table 3-1).

However, these results should not be considered definitive, as we were unable to detect

Silver Carp or Bighead Carp DNA in reaches where these fish have been documented with conventional gears (Chapter 2).

Failure to capture target DNA was likely influenced by multiple factors such as the high turbidity of the Kansas River (Thorp and Mantovani 2005) and differing seasonal temperatures. Filtration of water samples yield higher concentrations of genetic material than centrifugation (Eichmiller et al. 2016). However, elevated turbidity in the

Kansas River during summer sampling events (Table 3-3) prevented the filtration of water samples by clogging collection filters. Some techniques to alleviate this issue have been explored such as centrifuging water samples (Williams et al. 2017) or using multiple filters for each sample and pooling the extraction product from each filter (Robson et al.

2016). Thus, we opted to centrifuge samples collected in the summer because using the 66 multiple filter method would have greatly increased the cost of the assay. We were able to filter water samples collected in the winter, yet these samples failed to yield Silver

Carp or Bighead Carp DNA. Possible explanations of these outcomes include reduced shedding rates of genetic material at colder water temperatures (Jo et al. 2019) as metabolic rates are highly governed by ambient water temperatures (Fry 1974;

Horodysky et al. 2015). For example, gastric evacuation rates of Flathead Catfish

Plyodictis olivaris were positively correlated with temperature during laboratory studies

(Horstman 2020). Additionally, Silver Carp exhibit downstream movements in the fall

(Coulter et al. 2016) possibly reducing the overall abundance of these fish in the Kansas

River at the time of sample collection.

The volume of water in the river at the time of sample collection influences concentration of aqueous DNA (Curtis et al. 2020). We have found no research investigating what volume of water researchers need to sample relative to the volume of water in either lotic or lentic systems. This information is vital for researchers and managers to have confidence in either positive or negative results and for standardization of eDNA techniques. The ratio of volume of water sampled (6 L per site) to the volume of water in the system (approximately 850 to 900 m3 second-1 in the summer and 150 m3 second-1 in the winter; Table 3-3) was very low in our own study. Our sample size was sufficient to detect target organisms at greater abundances in Segment 1. However, larger sample sizes (either larger sample volume or increased number of samples) are likely necessary for positive detection of Silver Carp and Bighead Carp in Segment 2

(Willoughby et al. 2016; Sepulveda et al. 2019) where relative abundance has been found 67 to be significantly lower (Chapter 2). The density of target organisms factors into detectability, where higher abundances of target organisms is correlated with higher concentrations of aqueous DNA (Lodge et al. 2012; Takahara et al. 2012; Pilliod et al.

2013; Klymus et al. 2015; Wilcox et al. 2016 Coulter et al. 2019, Jo et al. 2019).

Additionally, we collected our pilot study samples near the confluence with the Missouri

River where downstream flow likely facilitated the accumulation of the greatest concentrations of genetic material. The lower abundance of Silver Carp in Segment 2 likely limited the accumulation of target genetic material relative to the pilot study samples, leading to undetectable concentrations of aqueous DNA.

Alternative PCR procedures such as quantitative PCR (qPCR) or digital droplet

PCR (ddPCR) are more sensitive than the conventional PCR methods used here (Nathan et al. 2014; Wilcox et al. 2016; Xia et al. 2018). Using the more sensitive PCR procedures in place of conventional PCR could increase detection rates when target DNA is low in concentration. However, qPCR and ddPCR currently have limited use because of operational complexity and availability (Nathan et al. 2014; Doi et al 2015; Xia et al.

2018). Additionally, ddPCR and qPCR are more expensive than conventional PCR, with qPCR being the most expensive of the three options (Nathan et al. 2014).

Continued monitoring for Silver Carp and Bighead Carp in Segment 3 is necessary for early detection and subsequent management action. Environmental DNA techniques could provide cost-effective and quick means of monitoring for these fishes.

However, eDNA techniques need refinement for application in turbid systems such as the

Kansas River for early detection of invasive species. 68

The detection of Black Carp in the Kansas Rivers warrants further investigation into their presence throughout the Missouri River basin. Additionally, investigation to confirm the presence of Black Carp in the Kansas River is needed to validate our eDNA results. Our results suggest sampling effort should be focused in Segment 1 near the confluence of the Kansas and Missouri Rivers.

Environmental DNA analyses are a powerful tool that have been shown to be useful for detecting fishes at low densities (e.g., Laramie et al. 2015; Pfleger et al. 2016).

As such, eDNA analyses have become increasingly common and will be a vital tool for detecting rare fishes or documenting the expansion of invasive species in the future.

Understanding the limitations of field and laboratory techniques, and how to overcome those limitations, is critical to improve the efficacy of eDNA methodology. Future research aimed at comparing eDNA use in varying environmental conditions is warranted. 69

Literature Cited Chick, J. H., D. K. Gibson-Reinemer, L. Soeken-Gittinger, and A. F. Casper. 2020. Invasive Silver Carp is empirically linked to declines of native sportfish in the Upper Mississippi River system. Biological Invasions 22:723-734. Conover, G., R. Simmons, and M. Whalen. 2007. Management and Control plan for Bighead, Black, Grass, and Silver Carps in the United States. Asian carp Working Group, Aquatic Nuisance Species Task Force, Washington D.C. 223 pp. Coulter, A. A., E. J. Bailey, D. Keller, and R. R. Goforth. 2016. Invasive Silver Carp movement patterns in the predominantly free-flowing Wabash River (Indiana, USA). Biological Invasions 18:471-485. Coulter, D. P., P. Wang, A. A. Coulter, G. E. Susteren, J. J. Eichmiller, J. E. Garvey, and P. W. Sorensen. 2019. Nonlinear relationship between Silver Carp density and their eDNA concentration in a large river. PLoS ONE e0218823. Curtis, A. N., J. S. Tiemann, S. A. Douglass, M. A. Davis, and E. R. Larson. 2020. High stream flows dilute environmental DNA (eDNA) concentrations and reduce detectability. Diversity and Distributions 00:1-14. Dean, Q. J. 2020. Population characteristics and movement of Blue Catfish in the Kansas River. Master’s thesis. University of Nebraska, Lincoln. DeGrandchamp, K. L., J. E. Garvey, and R. E. Colombo. 2008. Movement and habitat selection by invasive Asian carp in a large river. Transactions of the American Fisheries Society 137:45-56. Doi, H., K. Uchii, T. Takahara, S. Matsuhashi, H. Yamanaka, and T. Minamoto. 2015. Use of droplet digital PCR for estimation of fish abundance and biomass in environmental DNA surveys. PLoS ONE e0122763. Eichmiller, J. J., L. O. Moller and P. W. Sorensen. 2016. Optimizing techniques to capture and extract environmental DNA for detection and quantification of fish. Molecular Ecology 16:56-68. Eitzmann, J. L., A. S. Makinster, and C. P. Paukert. 2007. Distribution and growth of blue sucker in a great plains river, USA. Fisheries Management and Ecology 14:255-262. Ficetola, G. F., C. Miaud., F. Pompanon, and P. Teberlet. 2008. Species detection using environmental DNA from water samples. Biological Letters 4:423-425. Freeze, M. and S. Henderson. 1983. Distribution and Status of the Bighead Carp and Silver Carp in Arkansas. North American Journal of Fisheries Management 2:197-200. Fry, F. E. J. 1974. Effects of the environment on animal activity. The University of Toronto Press, Toronto, Ontario, Canada. 70

Hayer, C.-A., J. J. Breeggemann, R. A. Klumb, B. D. S. Graeb, and K. N. Bertrand. 2014. Population characteristics of Bighead and Silver Carp on the Northwestern front of their North American invasion. Aquatic Invasions 9:289-303. Horodysky, A. Z., S. J. Cooke, R. W. Brill. 2015. Physiology in the service of fisheries science: why thinking mechanistically matters. Reviews in Fish Biology and Fisheries 25:425-447. Horstman, Z. 2020. The effects of temperature on evacuation rates and absorption efficiency of Flathead Catfish. Master’s thesis. University of Nebraska, Lincoln. Irons, K. S., G. G. Sass, M. A. McClelland, and J. D. Stafford. 2007. Reduced condition factors for two native fish species coincident with invasion of non-native Asian carps in the Illinois River, U.S.A. Is this evidence for competition and reduced fitness? Journal of Fish Biology 71:258-273. Jerde, C. L., W. L Chadderton, A. R. Mahon, M. A. Renshaw, J. Corush, M. L. Budney, S. Mysorekar, and D. M. Lodge. 2013. Detection of Asian carp DNA as part of a Great-Lakes basin wide surveillance program. Canadian Journal of Fisheries and Aquatic Sciences 70:522-526. Jerde, C. L., A. R. Mahon, W. L. Chadderton, D. M. Lodge. 2011. “Sight-unseen” detection of rare aquatic species using environmental DNA. Conservation Letters. Jo, T., H. Murakami, S. Yamamoto, R. Masuda, and T. Minamoto. 2019. Effect of water temperature and fish biomass on environmental DNA shedding, degradation, and size distribution. Ecology and Evolution 9:1135-1146. Klymus, K. E., C. A. Richter, D. C. Chapman, and C. Paukert. 2015. Quantification of eDNA shedding rates from invasive Bighead Carp Hypophthalmichthys nobilis and Silver Carp Hypophthalmichthys molitrix. Biological Conservation 183:77- 84. Kolar, C. S., W. R. Courtenay, and L. G. Nico. 2010. Managing undesired and invading fishes. Pages 213-259 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Kolar, C. S. and D. M. Lodge. 2002. Ecological predictions and risk assessment for alien fishes in North America. Science 298:1233-1236. Laramie, M. B., D. S. Pilliod, and C. S. Goldberg. 2015. Characterizing the distribution of an endangered salmonid using environmental DNA analysis. Biological Conservation 183:29-37. Lodge, D. M., C. R. Turner, C. L. Jerde, M, A. Barnes, L. Chadderton, S. P. Egan, J. L. Feder, A. R. Mahon, and M. E. Pfrender. 2012. Conservation in a cup of water: estimating biodiversity and population abundance from environmental DNA. Molecular Ecology 21:2555-2558. 71

Mahon, A. R., C. L. Jerde, M. Galaska, J. L. Bergner, W. L. Chadderton, D. M. Lodge, M. E. Hunter, and L. G. Nico. 2013. Validation of eDNA surveillance sensitivity for detection of Asian carps in controlled field experiments. PLoS ONE 8:e58316. Makinster, A. S., and C. P. Paukert. 2008. Effects and utility of minimum length limits and mortality caps for Flathead Catfish in discrete reaches of a large prairie river. North American Journal of Fisheries Management 28:97-108. Moy, P. B., I. Polls, and J. M. Dettmers. 2011. The Chicago Sanitary and Ship Canal Aquatic Nuisance Species Barrier. Pages 121-138 in D. C. Chapman and M. H. Hoff, editors. Invasive Asian carps in North America. American Fisheries Society, Symposium 74, Bethesda, Maryland. Nathan, L. M., M. Simmons, B. J. Wegleitner, C. L. Jerde, and A. R. Mahon. 2014. Quantifying environmental DNA signals for aquatic invasive species across multiple detection platforms. Environmental Science and Technology 48:12800- 12806. Nico, L. G. and H .J. Jelks. 2011. The black carp in north America: an update. American Fisheries Society Symposium 74:89-104. Nico, L. G., and M. E. Neilson. 2018. Mylopharyngodon piceus (Richardson, 1846); USGS Nonindigenous Aquatic Species Database, Gainesville, FL. Available: https://nas.er.usgs.gov/queries/FactSheet.aspx?speciesID=573. Revision date: 8/6/2018. Paukert, C. A. and D. L. Galat. 2010. Warmwater rivers. Pages 699-736 in W.A Hubert and M.C Quist, editors. Inland Fisheries Management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Peters, L. M., M. A. Pegg, and U. G. Reinhardt. 2006. Movements of adult radio-tagged Bighead Carp in the Illinois River. Transactions of the American Fisheries Society 135:1205-1212. Pfleger, M. O., S. J. Rider, C. E. Johnston, and A. M. Janosik. 2016. Saving the doomed: using eDNA to aid in detection of rare sturgeon for conservation (Acipenseridae). Global Ecology and Conservation 8:99-107. Pilliod, D. S., C. S. Goldberg, R. S. Arkle, and L. P. Waits. Estimating occupancy and abundance of stream amphibians using environmental DNA from filtered water samples. Canadian Journal of Fisheries and Aquatic Sciences 70:1123-1130. Porto, L. M., R. L. McLaughlin, and D. L. G. Noakes. 1999. Low-head barrier dams restrict the movement of fishes in two lakes in Ontario streams. North American Journal of Fisheries Management 19:1028-1036. Quist, M. C., and C. S. Guy. 1999. Spatial variation in population characteristics of the Shovelnose Sturgeon in the Kansas River. The Prairie Naturalist 31:65-74. 72

Robson, H. L. A., T. H. Noble, R. J. Saunders, S. K. A. Robson, D. W. Burrows, and D. R. Jerry. 2016. Fine-tuning for the tropics: application of eDNA technology for invasive fish detection in tropical freshwater ecosystems. Molecular Biology 16:922-932. Sampson, S. J., J. H. Chick, and M. A, Pegg. 2009. Diet overlap among two Asian carp and three native fishes in backwater lakes on the Illinois and Mississippi rivers. Biological Invasions 11:483-496. Sass, G. S., C. Hinz, A. C. Erickson, N. N. McClelland, M. A. McClelland, and J. M. Epifanio. 2014. Invasive Bighead and Silver Carp effects on zooplankton communities in the Illinois River, Illinois USA. Journal of Great Lakes Research 40:911-921. Sepulveda, A. J., J. Schabacker, S. Smith, R. Al-Chokhachy, G. Luikart, and S. J. Amish. 2019. Improved detection of rare, endangered and invasive trout in using a new large-volume sampling method for eDNA capture. Environmental DNA 1:227- 237. Sparks, E. R., T. L. Barkley, S. M. Creque, J. M. Dettmers, and K. M. Stainbrook. 2011. Evaluation of an electric fish dispersal barrier in the Chicago Sanitary and Ship Canal. Pages 139-162 in D. C. Chapman and M. H. Hoff, editors. Invasive Asian carps in North America. American Fisheries Society, Symposium 74, Bethesda, Maryland. Takahara, T., T. Minamoto, H. Yamanaka, H. Doi, and Z. Kawabata. 2012. Estimation of fish biomass using environmental DNA. PLoS ONE e35868. Thomsen, P. F., J. Kielgast. L. L. Iversen, C. Wiuf, M. Rasmussen, M. T. P. Gilbert, L. Orlando, and E. Willerslev. Monitoring endangered freshwater biodiversity using environmental DNA. Molecular Biology 21:2565-2573. Thorp, J. H. and S. Mantovani. 2005. Zooplankton of turbid and hydrologically dynamic prairie river. Freshwater Biology 50:1474-1491. Tripp, S., R. Brooks, D. Herzog, and J. Garvey. 2014. Patterns of fish passage in the upper Mississippi River. River Research and Application 30:1056-1064. Vander Zanden, J. M. and J. D. Olden. 2008. A management framework for preventing the secondary spread of aquatic invasive species. Canadian Journal of Fisheries and Aquatic Sciences 65:1512-1522. Wilcox, T. M., K. S. McKelvey, M. K. Young, A. J. Sepulveda, B. B. Shepard, S. F. Jane, A. R. Whiteley, W. H. Lowe, and M. K. Schwartz. 2016. Understanding environmental DNA detection probabilities: a case study using stream-dwelling char Salvelinus fontinalis. Biological Conservation 194:209-216. 73

Williams, K. E., K. P. Huyvaert, and A. J. Piaggio. 2017. Clearing muddied waters: capture of environmental DNA from turbid waters. PLoS ONE e0179282. Willoughby, J. R., B. K. Wijayawardena, M. Sundaram, R. K. Swihart, and J. A. Dewoody. 2016. The importance of including imperfect detection models in eDNA experimental design. Molecular Ecology 16:837-844. Xia, Z., M. L. Johansson, Y. Gao, L. Zhang, G. D. Haffner, H, J, MacIsaac, and A. Zhan. 2018. Conventional versus real-time quantitative PCR for rare species detection. Ecology and Evolution 8:11799-11807. 74

Table 3-1: Detection rate (Dr) of Silver Carp, Bighead Carp, and Black Carp at each sampling site in the Kansas River, and from the pilot study (P) near the confluence with the Missouri River using both centrifugation procedures and filtration procedures. Dashes indicate samples from sites that were not processed using those procedures.

Dr Centrifuge (%) Dr Filter (%) Silver Bighead Black Silver Bighead Black Sites Carp Carp Carp Carp Carp Carp P 100 0 100 - - -

Segment 1 - - - 0 0 0 1 2 - - - 0 0 0 3 - - - 0 0 0 4 0 0 0 - - - 5 0 0 0 - - - Segment 6 0 0 0 - - - 2 7 - - - 0 0 0 8 - - - 0 0 0 9 0 0 0 0 0 0 10 0 0 0 - - - Segment 11 0 0 0 - - - 3 12 0 0 0 - - - 13 0 0 0 - - -

75

Table 3-2: False negative rate (Fr) for Silver Carp and Bighead Carp in all samples collected in the Kansas River below Bowersock Dam, located in Lawrence, Kansas.

# of Samples Positive Negative Fr (%) Silver Carp Centrifugation 21 3 18 86 Filtration 36 0 36 100 Bighead Carp Centrifugation 21 0 21 100 Filtration 36 0 36 100

76

Table 3-3: Mean daily discharge (m3 second-1), and mean daily turbidity, measured in

Formazin Nephelometric Units, in the Kansas River on eDNA sample collection days.

Data were collected at the Lake Quivira gaging station located near Edwardsville, KS

(USGS Gage 06892518).

Discharge (m3 second-1) Turbidity (FNU) August 20, 2019 855 230 August 23, 2019 898 208 February 17, 2020 148 8.4

77

Figure 3-1: Kansas River study area where Segment 1 (24 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (61 rkm) is between the Johnson County Weir and Bowersock Dam, and Segment 3 (57 rkm) is between Bowersock Dam and the Topeka Weir. White stars indicate water sample collection sites with site numbers next to them. 78

CHAPTER 4: ENVIRONMENTAL HISTORY OF SILVER CARP IN THE LOWER KANSAS RIVER, KANSAS: INSIGHT FROM OTOLITH MICROCHEMISTRY ANALYSIS

J. P. WERNER*, Q. J. DEAN, M. A. PEGG, and M. J. HAMEL

Abstract Invasive Silver Carp Hpophthalmichthys molitrix have established populations throughout the Missouri River basin, including in the Kansas River. Understanding the spatial extent under which these invasive fish function in large, open river systems is crucial to inform management efforts. Tributaries such as the Kansas River may play a vital role in the life-cycle of Silver Carp in the Missouri River basin as the main-stem

Missouri River has undergone a multitude of alterations, creating a channel with greater mean depths and velocities. Here, we used otolith microchemistry of Silver Carp from the

Kansas River to reconstruct environmental histories as means to assess the proportions of resident and transient individuals. Silver Carp within the Kansas River were predominantly residents (adults = 54%; juveniles = 65%) with the majority of reproduction coming from within the Kansas River itself. Therefore, removal efforts in the Kansas River may be effective means of managing this invasive fish. Transient individuals exhibited short durations of signatures indicative of the Missouri River (mean percent of data points for adults = 10% and juveniles = 36%), suggesting movements into the Missouri River were brief. These results highlight the importance of connectivity of tributary habitat among large rivers and provides important information for invasive species management. 79

Introduction Silver Carp Hypophthalmichthys molitrix are an invasive species from eastern

Asia that invaded U.S.A. waterways in the early 1980’s (Freeze and Henderson 1982;

Conover et al. 2007; Lu et al. 2020). Since their introduction, they have expanded their range to encompass the majority of the Mississippi River Basin (Conover et al. 2007), including the lower Missouri River drainage where they have expanded as far North as

North Dakota (Hayer et al. 2014). The spatial extent under which Silver Carp function in the Missouri River drainage is unclear because of the open nature and connectivity of this large river system. The lower Missouri River (below Gavins Point Dam, SD) is devoid of dams leaving approximately 1,305 river kilometers (rkm) and numerous connections to tributaries open to immigration and emigration of Silver Carp. These open corridors, coupled with their ability for long, longitudinal movements (DeGrandchamp et al. 2008;

Coulter et al. 2016), has aided in range expansion into and throughout the lower Missouri

River drainage.

Modifications for flood control and to maintain a navigable channel (i.e., wing dikes, levies, and bank stabilization structures) are extensive throughout the lower

Missouri River. These modifications created greater mean depths and velocities while also limiting lateral connectivity with the river floodplain (Galat et al. 1998; Pegg et al.

2003; Steffensen and Mestl 2016). Consequently, limited optimal habitat remains for

Silver Carp in the main stem of the Missouri River because they tend to prefer areas with lower velocities (DeGrandchamp et al. 2008; Calkins et al 2012). Tributaries to the

Missouri River, and limited habitat behind wing dikes, may provide refuge habitat for

Silver Carp seeking to escape the high velocity flows of the Missouri River (Kolar et al. 80

2005). Additionally, tributaries may act as stepping-stones for longitudinal movement throughout the basin.

Longitudinal connectivity between populations influences biological processes

(Pegg and Chick 2010) such as gene flow. Transient Silver Carp within the population facilitate this connectivity between populations. Silver Carp exhibit individual based movement patterns where some individuals within a given population are transient and others are resident (Coulter et al. 2016; Prechtel et al. 2018). Dichotomy in individual based movement patterns has been observed in other riverine fishes such as Common

Carp Cyprinus Carpio (Butler and Wahl 2010), many Salmonids (Rodríguez 2002) and

Guadalupe Bass Micropterus treculii (Perkin et al. 2010). This strategy promotes dispersal and the colonization of new areas while also ensuring that some individuals stay in suitable habitat (Coulter et al. 2016). Discerning the proportions of transient and resident individuals within Silver Carp populations can provide insight for management action (Prechtel et al. 2018). For example, populations comprised mostly of resident individuals would be ideal for removal efforts. Timing removal efforts to coincide with periods of reduced movement distances (i.e., summer months) could be effective means for removing transient individuals (Prechtel et al. 2018).

Analysis of stable isotopes and trace elemental composition of otoliths as natural tags is a useful tool for retrospectively identifying the environmental history of riverine fishes (Zeigler and Whitledge 2010). Strontium (Sr) and Barium (Ba) – in ratios to

Calcium (Ca) – are two common elements used in microchemistry analyses (Whitledge et al. 2007; Zeigler and Whitledge 2010; Crook et al. 2013; Carlson et al. 2016, Whitledge 81 et al. 2018). For example, Sr:Ca ratios were used to identify natal origins of Silver Carp captured in the Illinois River to the Illinois River itself, the Missouri River, or the middle

Mississippi River (Norman and Whitledge 2015). Additionally, Sr:Ca ratios were used to identify origins of Silver Carp captured in urban Chicago fishing ponds to the Illinois

River (Love et al. 2019). Here, we used otolith microchemistry to reconstruct the environmental history of Silver Carp captured in the Kansas River to determine the proportion of resident and transient individuals. Additionally, we aimed to quantify tributary versus main-stem Missouri River occupancy durations of transient Silver Carp within the Kansas River. These data will provide insight into movement patterns of Silver

Carp between the Missouri River and the Kansas River habitats and help to determine if removal efforts would be effective at reducing abundance of Silver Carp in the Kansas

River.

Methods

Study Area

The Kansas River is a large tributary to the Missouri River (Figure 4-1). Its origins are at the confluence of the Smoky Hill and Republican Rivers (Quist and Guy

1999) in North-Central Kansas and flows easterly 274 kilometers (Makinster and Paukert

2008) to its confluence with the Missouri River near Kansas City, Kansas. Discharge is controlled by 18 federal reservoirs and over 13,000 small impoundments (Quist et al.

1999). The main-stem of the Kansas River has three major barriers; the Topeka Weir in

Topeka, Kansas at river-kilometer (rkm) 141, Bowersock Dam in Lawrence, Kansas at rkm 83, and the Johnson County Weir in Edwardsville, Kansas at rkm 27 (Figure 4-1). 82

Bowersock Dam functions as a hydropower dam and is classified as a low-head dam

(Quist and Guy 1999) that can impede upstream longitudinal movements of riverine fishes (Eitzman et al. 2007; Dean 2020), including Silver Carp (Chapter 2).

Our study area was the lower half of the Kansas River from the Topeka Weir, in

Topeka, Kansas to the Confluence with the Missouri River. Additionally, we included a

22 rkm segment of the Missouri River and the Wakarusa River from its confluence with the Kansas River to the Clinton Lake Dam for otolith and water data collection. We divided the Kansas River into three distinct segments associated with three barriers on the main stem of the Kansas River. Segment 1 is between the confluence with the Missouri

River and the Johnson County Weir (27 rkm), Segment 2 is between the Johnson County

Weir and Bowersock Dam (56 rkm), and Segment 3 is between Bowersock dam and the

Topeka Weir (58 rkm). The Wakarusa River joins the Kansas River in Segment 2 near

Eudora, KS (Figure 4-1).

Microchemistry Data Collection

We collected water samples in autumn 2018, winter 2019, and summer 2019 to assess spatiotemporal variations in water trace element concentrations (Ciepiela and

Walters 2019). We gathered water samples across the Kansas River (Topeka Weir to the confluence with the Missouri River; n = 25 samples), Wakarusa River (below Clinton

Lake to the confluence with the Kansas River; n = 5 samples), and Missouri River (16 km upstream of the confluence to 7 km below the confluence with the Kansas River; n = 7 samples). We collected water samples using a syringe filtration technique from 13 sites across the three river systems (Kansas River = 9 sites, Wakarusa River = 2 sites, Missouri 83

River = 2 sites) (Figure 4-1) with a 250ml polyethylene bottle. We rinsed the bottle a minimum of three times before collecting each water sample. We then rinsed a syringe filter and used it to filter 15ml of water into a cleaned and rinsed collection bottle. We stored the sample bottles in a cool, dark location until we sent them to the lab for analysis. We analyzed samples for Strontium (Sr), Barium (Ba), Magnesium (Mg),

Calcium (Ca), Sodium (Na), and Manganese (Mn) at the University of Southern

Mississippi’s Center for Trace Analysis. Data were reported as the molar concentration for each trace element and converted to element:Ca (mmol/mol) ratios (e.g., Sr:Ca,

Ba:Ca, and Mg:Ca).

We collected Juvenile and adult Silver Carp from Segments 1 and 2 from the

Wakarusa River in May-August of 2018 and 2019. No Silver Carp were collected above

Bowersock Dam in Segment 3 (Chapter 2). We used a combination of electrofishing gears and mini-fyke nets throughout the reach to collect Silver Carp (e.g., Chapter 2). We extracted both lapilli otoliths from a minimum of 25 Silver Carp per segment per month in

2018 and 2019. We analyzed the first 25 otoliths collected from each segment in each month in 2018. In 2019, we selected otoliths that were devoid of cracks and other imperfections. In total, we selected 300 Silver Carp otoliths for microchemistry analysis. We collected otoliths from only adult Silver Carp (>400 mm) in 2018, whereas in 2019 we collected otoliths from both juvenile Silver Carp (<400 mm) and adult Silver Carp. The majority of otoliths from juvenile Silver Carp were collected in Segment 1 (Table 4-2) because juveniles were rare in

Segment 2 (Chapter 2). Additionally, we aimed to select otoliths for microchemistry analysis based on spatiotemporal variations on when they were collected to monitor for shifts in trace element signatures over both time and space. We extracted otoliths by making an incision 84 through the top of the skull into the cranial cavity and collected the otoliths using non- metallic tweezers. Then, we cleaned the otoliths of flesh and placed them in 2 ml polyethylene vials until they were prepared for ablations in the lab.

We washed the otoliths with deionized water and allowed them to dry for 24 hours. We then embedded the otoliths in epoxy (Epoxicure Epoxy Resin and Hardener,

Beuhler Inc., Lake Bluffs, Illinois) and sectioned them across the transverse plane through the nucleus using a Buehler IsoMet low speed saw. We sanded otolith sections using 1,500 and 3,000 grit sandpaper and polished them to reveal annuli using 3µm lapping paper. We then rinsed the sectioned otoliths in deionized water, adhered them to microscope slides with double sided tape, and stored them until microchemistry analysis. We analyzed the trace element composition of the otoliths using a Thermo X-Series2 ICPMS coupled with a

Teledyne-CETAC Technologies LSX-266 laser ablation system. Ablations (beam diameter =

20μm, scan rate = 5μm/sec., laser pulse rate = 5hz) began approximately 100µm on one side of the nucleus, ablated completely thought the nucleus, to the adjacent edge of the otolith. We analyzed a standard developed by the U.S. Geological Survey (USGS) (MACS-3, CaCO3 matrix) every 15-20 samples to adjust for instrument drift using procedures outlined by

Whitledge et al. (2018). Otolith microchemistry data were converted to molar concentrations for each element and reported as element:Ca (µmol/mol) ratios (e.g., Sr:Ca, Ba:Ca, and

Mg:Ca) using calcium as the internal standard and the stoichiometric concentration of calcium in aragonite Calcium Carbonate (CaCO3) (Whitledge et al. 2018).

We primarily focused on Sr:Ca ratios to classify the environmental history of

Silver Carp. Sr:Ca ratios are commonly used in microchemistry analysis of Silver Carp otoliths (e.g., Norman and Whitledge 2015, Whitledge et al. 2018, Love et al. 2019). 85

Specifically, lapilli otoliths are analyzed for their Sr:Ca ratios because they are usually comprised of the aragonite polymorph of CaCO3 (Whitledge et al. 2018). Aragonite has a higher affinity for Sr compared to other polymorphs such as vatarite and calcite

(Campana 1999; Melancon et al. 2005; Pracheil et al. 2019). A combination of Sr:Ca,

Ba:Ca, and Mg:Ca ratios were used as an indicator for vatarite otoliths (Mg:Ca >

400µmol/mol, Ba:Ca < 4µmol/mol, and Sr:Ca < 100µmol/mol) (Whitledge et al. 2018). We did not use Mg any further in the reconstruction of environmental history because metabolic processes more heavily regulate Mg than Sr or Ba. Magnesium contributes to the phosphorylation of enzymes and is a co-factor in adenosine triphosphate (ATP).

Additionally, the hydrated and dehydrated forms of Mg have largely different radiuses than Ca+ and are likely randomly trapped in the crystal lattice (Thomas et al. 2017; Hüssy et al. 2020). Conversely, Sr and Ba have similar radii to Ca and compete for Ca binding sites in the crystal lattice (Hüssy et al. 2020).

Data Analysis

We used otoliths collected from Silver Carp captured from Segment 2 and the

Wakarusa River in 2018 to characterize the relationship between water and otolith microchemistry signatures in the Kansas and Wakarusa Rivers. During low flow events – such as those in 2018 – the Johnson County Weir was likely a barrier to movement between Segment 1 and Segment 2 (Chapter 2; Dean 2020). Isolation of Silver Carp in

Segment 2 for an extended period (e.g., >3 months) likely facilitated trace elemental equilibrium in the signatures. Additionally, movement rates of Silver Carp are lower during low flow events (DeGrandchamp et al. 2008; Calkins et al. 2012; Coulter et al. 86

2016; Prechtel et al. 2018), likely limiting movement between the main-stem Kansas

River and its tributaries. We characterized the relationship between water and otolith microchemistry signatures in the Missouri River using ablation data from Silver Carp captured in isolated backwaters of the Missouri River (n = 13). Data were provided by the

Center for Fisheries, Aquaculture, and Aquatic Sciences at Southern Illinois University –

Carbondale. We isolated trace elemental ratio data at the edge of the selected otoliths

(~30 µm) to characterize the relationship between water and otolith signatures in the

Kansas, Missouri, and Wakarusa Rivers (Norman and Whitledge 2015; Spurgeon et al

2018).

We grouped water samples and otoliths by river of sample collection and assessed differences using univariate and multivariate methods. Tests of normality (i.e., Shapiro-

Wilks tests) indicated water and otolith microchemistry data deviated from normality, thus we proceeded with non-parametric tests. We used a permutated multivariate analysis of variance (perMANOVA) to examine the differences in otolith and water trace elemental signatures between river systems, adjusting the P-value using the Bonferroni method for pair-wise comparisons. We then used a Kruskal-Wallis test followed by a post hoc Dunn’s test to examine pair-wise differences in water and otolith trace elemental signatures between river systems, adjusting the P-value for multiple comparisons using the Holmes method.

We used a recursive partitioning modelling approach to separate trace elemental signatures in Silver Carp otoliths between river systems. We built the recursive partitioning tree using the rpart package (Thernau et al. 2019) in program R (R Core 87

Team 2020). Recursive partitioning trees aim to split the data into homogenous groups to increase the homogeneity of the elements (i.e., trace elemental ratios) within groups (i.e., rivers) (Dinov 2018). We used a splitting criterion based on the Gini impurity index (e.g.,

Spurgeon et al. 2018) and selected the tree within one standard error to the tree with the lowest cross-validated error (Spurgeon et al. 2018; Thernau et al. 2019). We then used the resulting partitioning tree to predict the trace elemental threshold that distinguishes each river. We classified individual Silver Carp as “transient” if they had signatures indicative of the Missouri River or “resident” if they lacked those signatures (e.g., Figure

4-5). We excluded Silver Carp with trace elemental signatures indicative of vatarite

CaCO3 otoliths or with extremely high Sr:Ca ratios (e.g. > 5,000µmol/mol) from the analysis.

We determined the proportion of transient adult and juvenile Silver Carp in

Segments 1 and 2 in both 2018 and 2019. Additionally, we determined the proportion of transient Silver Carp that were male and female to examine sex-specific movement patterns. We then isolated ablation data from the nucleus of the otolith to determine the proportion of Silver Carp in the Kansas River with natal origins from the Missouri River.

We also enumerated movement events between the Missouri River and other water bodies. We classified movement events as a shift in trace element data to signatures indicative of the Missouri River at any point along the ablated transect.

We also aimed to quantify the percent of time each transient fish spent in the

Missouri River versus other water bodies in the drainage to gain insight on habitat use by

Silver Carp in the basin. We calculated the percent of Sr:Ca data points indicative of the 88

Missouri River (i.e., percent of points in each spike) for each transient fish. We then plotted this value for each individual by its total length (TL).

Results

Water Chemistry

Multivariate comparisons among rivers indicated water trace elemental signatures

(i.e., the combination of Sr:Ca and Ba:Ca ratios) differ between the Kansas River and

2 Wakarusa River (F = 74.3, r = 0.726, P = 0.003), Kansas River and Missouri River (F =

2 2 24.1, r = 0.445, P = 0.003), and Wakarusa River and Missouri River (F = 41.1, r =

0.804, P = 0.003). Univariate comparisons indicate water Sr:Ca ratios (χ2 = 22.5, P <

0.001) and Ba:Ca ratios (χ2 = 13.9, P < 0.001) differ between rivers. Pair-wise comparisons revealed that water Sr:Ca ratios were higher in the Kansas River than both the Missouri River (Z = 31.14, P = 0.003) and the Wakarusa River (Z = 4.09. P < 0.001).

However, water Sr:Ca ratios were similar between the Wakarusa River and the Missouri

River (Z = 1.13, P = 0.26). Water Ba:Ca ratios were similar between the Kansas River and Missouri River (Z = -1.6, P = 0.111). Water Ba:Ca ratios were lower in the Wakarusa

River than either the Kansas River (Z = 3.01, P = 0.005) or the Missouri River (Z = 3.68,

P < 0.001) (Figure 4-2).

Otolith Chemistry

We used a total of 73 adult Silver Carp with a mean total length of 671 mm (sd =

46 mm) to classify the relationship between water and otolith microchemistry in the

Kansas River (n = 59) and Wakarusa River (n = 14). Additionally, we used this subset of data to train and test the recursive partitioning tree for the following, larger analysis. 89

Multivariate tests among rivers indicated otolith trace elemental signatures did not differ between the Kansas River and Wakarusa River (F = 7.47, r2 = 0.095, P = 0.027), but did differ between the Kansas River and Missouri River (F = 84.6, r2 = 0.547, P = 0.003), and

Wakarusa River and Missouri River (F = 75.5, r2 = 0.751, P = 0.003). Univariate tests indicate trace elemental signatures differed among rivers in both Sr:Ca ratios (χ2 = 38.8, P

< 0.001) and Ba:Ca ratios (χ2 = 34.5, P < 0.001). Pair-wise comparisons revealed Otolith

Sr:Ca ratios were the higher in the Missouri River than either the Kansas River (Z = -

5.05, P < 0.001) or the Wakarusa River (Z = 2.65, P = 0.008). Additionally, otolith Sr:Ca ratios were higher in the Kansas River than the Wakarusa River (Z = 6.06, P < 0.001).

Otolith Ba:Ca ratios were higher in the Missouri River than either the Kansas River (Z = -

5.86, P < 0.001) or the Wakarusa River (Z = 3.48, P < 0.001). However, otolith Ba:Ca ratios did not differ between the Kansas River and Wakarusa River (Z = -1.27, P = 0.127)

(Figure 4-3).

The recursive partitioning tree correctly classified 92% of fish collected in the

Kansas River, 50% in the Missouri River, and 0% in the Wakarusa River. However, all fish collected from the Wakarusa River were classified as fish from the Kansas River

(Table 4-1). Thus, our model could only be used to distinguish between the Kansas River and the Missouri River (Sr:Ca > 2,082µmol/mol was indicative of the Missouri River)

(Figure 4-4). Ba:Ca was the most influential variable in our model (variable importance score = 57) followed by Sr:Ca (variable importance score = 43). The inclusion of Ba:Ca in the model did not refine the model enough to be able to further distinguish between the

Wakarusa River and the Kansas River and between the Wakarusa River and the Missouri 90

River. Additionally, Ba:Ca data were more erratic with multiple unreliable data points across all individuals while the Sr:Ca data were more consistent. Therefore, we primarily used Sr:Ca ratios in environmental history reconstruction.

We reconstructed environmental histories of 276 (n = 239 adult; n = 37 juvenile)

Silver Carp with approximately 46% of adults and 35% of juveniles classified as transient individuals. The proportion of transient individuals among sampling years and segment of capture for adult Silver Carp was consistently 45%-49%, except in Segment 1 during

2019 where the proportion of transients was approximately 22%. Juvenile Silver Carp were predominantly residents in 2019 with approximately 65% of individuals sampled lacking trace elemental ratios indicative of the Missouri River (Table 4-2). About 57% (n

= 37) of transient fish we identified gender for were male, and approximately 43% (n =

28) were female.

Approximately 17% (n = 19) of transient adult Silver Carp had natal origin signatures indicative of the Missouri River, 10 of which were captured in Segment 1 and the remaining 9 were captured in Segment 2. Approximately 46% (n = 6) of transient juvenile Silver Carp had natal origin signatures from the Missouri River. Five were captured in Segment 1 and one was captured in Segment 2. Overall, approximately 9% (n

= 25) of all fish sampled for microchemistry analysis had natal origins predicted to be from the Missouri River (Table 4-3).

A single movement event into the Missouri River was most common for both adult (n = 82) and juvenile (n = 10) transient Silver Carp. Two movement events occurred less often for adults and juveniles. Approximately 30% of transient adults (n = 91

24) and juveniles (n = 3) exhibited two movement events. Three movement events were exceedingly rare in adults (n = 3) while no juveniles captured exhibited three movement events (Table 4-3).

Transient juvenile Silver Carp exhibited a greater percent of points above the

Missouri River threshold (i.e., percent of points in each spike) than transient adult Silver

Carp. The percent of points above the Missouri River threshold for adults was less than

30% for the majority of the individuals, ranging from approximately 2% to 36% of points. Juveniles where more erratic, ranging from approximately 10% to 71% of points.

Overall, there was a negative relationship between the percent of points above the

Missouri River threshold and total length (Figure 4-6).

Discussion

Examination of otolith trace elemental signatures may be a useful tool in reconstructing environmental histories and predicting natal origins of Silver Carp in the

Kansas River and Missouri River systems. Our study indicated the population within the

Kansas River is comprised of predominantly residential individuals, having a consistent signature indicative of the Kansas River throughout the life span of the fish. Therefore, recruitment from within the Kansas River system is regularly occurring without connectivity to additional river systems. Segment 1 may be of particular importance for reproduction as the vast majority of juvenile Silver Carp were documented in this reach

(Chapter 2). Segment 1 has an average depth less than 1.5m and is characterized by low velocity flows (Eitzmann and Paukert 2010). These habitat conditions are typically where 92 age-0 Silver Carp are found at higher densities (e.g., Haupt and Phelps 2016) and are habitats commonly used as nursery grounds (Kolar et al. 2005, Conover et al. 2007).

Our data show that about 20% of all transient adult and juvenile Silver Carp in the

Kansas River had natal origin signatures indicative of the Missouri River. These results are noteworthy because Deters et al. (2013) documented the highest egg densities within the main-stem Missouri River while tributaries (e.g., Lamine River, Bonne Femme

Creek, Perch Creek, Moniteau Creek, Moreau River, and Osage River) had little egg production. However, Camacho et al. (2020) documented egg production in tributaries to the upper Mississippi River. This provides evidence that tributary habitats can be used for reproduction and is likely a function of select habitat availability. Therefore, it is likely that select tributaries throughout the Missouri River basin – such as the Kansas River – may be sources of reproduction and recruitment for Silver Carp. Identifying these sources throughout the basin would provide information as to where control efforts should be focused. Additionally, these data could reveal source-sink dynamics between the main stem Missouri River and adjacent systems.

Transient juvenile and adult Silver Carp exhibited trace elemental signatures indicative of the Missouri River for relatively short durations (e.g., Figure 4-5). These results indicate that transient Silver Carp occupied the Missouri River for brief periods of time compared to other systems. Brief occupancy in the Missouri River is likely influenced by the lack of optimal habitat available for Silver Carp. Additionally, higher velocity flows in the Missouri River (e.g., Pegg et al. 2003) facilitates the lack of buildup of autotrophic biomass and reduces residence time (Hosen et al; 2019), limiting food 93 availability for Silver Carp in this system. Areas with lower velocities, like the Kansas

River and areas behind wing dikes, may provide habitat with higher resource availability as well as refuge from the swift flows within the main channel (e.g., DeGrandchamp et al. 2008; Calkins et al. 2012; Coulter et al. 2016).

A few of the transient Silver Carp we analyzed had trace elemental signatures that indicated multiple movement events through the Missouri River. These results suggest the Missouri River may function more as a movement corridor for Silver Carp as they migrate between areas of suitable habitat. Further research on movement patterns of

Silver Carp throughout the Missouri River basin is needed to test this hypothesis.

Movement events into the Missouri River are likely induced by a variety of factors. For example, increased movement rates as a response to a rise in river flood stage has been documented for adult Silver Carp (Peters et al. 2006; DeGrandchamp et al. 2008; Coulter et al. 2016). Increased movement rates could facilitate movement events into the

Missouri River as transient individuals seek new habitats (Prechtel et al. 2018).

Additionally, broad scale upstream movements occurring in the spring typically occur as

Silver Carp stage for spawning events (Coulter et al. 2016). Brief forays into the Missouri

River could occur as Silver Carp seek suitable spawning habitat located in the Missouri

River itself or other tributaries in the basin.

We can distinguish habitat use between the Kansas River and Missouri River because of differences in water and otolith trace elemental signatures. However, otolith trace elemental signatures of the Wakarusa River and Kansas River were too similar to distinguish use between these two water bodies, even with the incorporation of multiple 94 trace elements (e.g., Sr:Ca and Ba:Ca). The paucity of water and otolith microchemistry data throughout the Missouri River basin limited our analysis to only between the Kansas and Missouri Rivers. Water chemistry signatures have been classified for the Platte River in Nebraska (e.g., Phelps et al. 2012; Spurgeon et al 2018) and throughout the main stem of the Missouri River below Gavins Point Dam , SD (e.g., Phelps et al. 2012; Norman and Whitledge 2015; Porreca et al. 2016; Spurgeon et al. 2018; Whitledge et al. 2018).

However, multitudes of other tributaries in the Missouri River drainage remain to be analyzed. Classification of these other tributaries will lead to insights in movement and recruitment sources of Silver Carp throughout the Missouri River basin.

Additional otolith trace elemental signatures need to be characterized throughout the Missouri River drainage (Hüssy et al. 2020). Our results demonstrate that the relationship between otolith and water signatures may not always be a positive linear relationship (Figures 4-2 and 4-3) and instead may resemble a logistic curve. Models built by Norman and Whitledge (2015) predicted otolith Sr:Ca ratios for Silver Carp captured from the Kansas River would be centered around 3,600 µmol/mol. However, we observed consistent values of approximately 1,500 µmol/mol (Figure 4-3). These differences could be explained by contamination, instrumental miscalibration, or procedural errors. However, the predicted otolith Sr:Ca values for the Wakarusa (~ 1,650

µmol/mol) is within the 95% confidence intervals of the observed values (Figure 4-3), indicating these errors were negligible. Biotic and abiotic factors such as salinity, temperature, oxygen, ontogeny, food and growth, and maturation can influence how trace elements are incorporated into the crystal lattice (Campana 1999; Norman and Whitledge 95

2015; Sturrock et al. 2015; Hüssy et al. 2020). One of these factors, or a combination of such, could have caused the negative relationship between otolith and water Sr:Ca values we observed in the Kansas River. Although these factors do not exert a strong influence on the biomineralization process of otolith formation (Hüssy et al. 2020), they could influence physiological processes governing trace element uptake and transport.

The Kansas River affords a unique opportunity for direct management and possible reduction of Silver Carp abundance because of the higher proportion of resident individuals. For example, removal efforts may be a viable option in the Kansas River, and should focus on Segment 1 because this reach is likely where reproduction is occurring (Chapter 2). Timing removal efforts to coincide with periods when Silver Carp are least active, such as during the late summer and early fall months (DeGrandchamp et al. 2008; Coulter et al. 2016), could have impacts throughout the Missouri River basin by removing a larger portion of transient individuals (Prechtel et al. 2018). Gears targeting all size groups, such as the electrified dozer trawl (Hammen et al. 2019; Chapter 2), should be used to maximize effort (Tsehaye et al. 2013), particularly during years when age-0 Silver Carp are confined to Segment 1 (e.g., Chapter 2).

Our results indicate that Silver Carp captured from the Kansas River tributary occupy waters adjacent to the mainstem Missouri River for most of their lives. Therefore, management of these invasive fish may be better suited focusing on the multitude of tributary habitats throughout the Missouri River drainage rather than the main stem

Missouri River. Future work investigating contributions of these tributary systems to the greater Missouri River population is essential to determine if management efforts 96 focusing on tributary systems will effectively diminish the greater Missouri River population. Our analysis indicates otolith microchemistry may provide effective means to investigate this relationship. 97

Literature Cited Butler, S. E., and D. H. Wahl. 2010. Common Carp distribution, movements, and habitat use in a river impounded by multiple low-head dams. Transactions of the American fisheries society 139:1121-1135. Calkins, H. A., S. J. Tripp, and J. E. Garvey. 2012. Linking silver carp habitat selection to flow and phytoplankton in the Mississippi River. Biological Invasions 14:949- 958. Camacho, C. A., C. J. Sullivan, and C. L. Pierce. Accepted. Invasive carp reproductive phenology in tributaries of the upper Mississippi River. North American Journal of Fisheries Management. Campana, S. E. 1999. Chemistry and composition of fish otoliths: pathways, mechanis,s and applications. Marine Ecology Progress Series 188:263-297. Carlson, A. K., M. J. Fincel, and B. D. S. Graeb. 2016. Otolith microchemistry reveals natal origins of walleyes in Missouri river reservoirs. North American Journal of Fisheries Management 36:341-350. Ciepiela, L. R. and A. W. Walters. 2019. Quantifying 87Sr/86Sr temporal stability and spatial heterogeneity for use in tracking fish movement. Canadian Journal of Fisheries and Aquatic sciences 76:928-936. Coulter, A. A., E. J. Bailey, D. Keller, and R. R. Goforth. 2016. Invasive Silver Carp movement patterns in the predominantly free-flowing Wabash River (Indiana, USA). Biological Invasions 18:471-485. Conover, G., R. Simmons, and M. Whalen. 2007. Management and Control plan for Bighead, Black, Grass, and Silver Carps in the United States. Asian carp Working Group, Aquatic Nuisance Species Task Force, Washington D.C. 223 pp. Crook, D. A., J. I. Macdonald, D. G. McNeil, D. M. Gilligan, M. Asmus, R. Maas, and J. Woodhead. 2013. Recruitment sources and dispersal of an invasive fish in a large river system as revealed by otolith chemistry analysis. Canadian Journal of Fisheries and Aquatic Sciences 70:953-963. DeGrandchamp, K. L., J. E. Garvey, and R. E. Colombo. 2008. Movement and habitat selection by invasive Asian carp in a large river. Transactions of the American Fisheries Society 137:45-56. Deters, J. E., D. C. Chapman, and B. McElroy. 2013. Location and timing of Asian carp spawning in the lower Missouri river. Environmental Biology of Fishes 96:617- 629. Dinov, I. O. 2018. Decision Tree Divide and Conquer Classification. Pages 307-344 in Data Science and Predictive Analytics. Springer International Publishing, Cham, Switzerland. 98

Eitzmann, J. L., A. S. Makinster, and C. P. Paukert. 2007. Distribution and growth of blue sucker in a great plains river, USA. Fisheries Management and Ecology 14:255-262. Eitzmann, J. L. and C. P. Paukert. 2010. Longitudinal differences in habitat complexity and fish assemblage structure of a great plains river. The American Midland Naturalist 163:14-32. Freeze, M. and S. Henderson. 1983. Distribution and Status of the Bighead Carp and Silver Carp in Arkansas. North American Journal of Fisheries Management 2:197-200. Galat, D. L., L. H. Fredrickson, D. D. Humburg, K. J. Bataille, J. R. Bodie, J. Dohrenwend, G. T. Gelwicks, J. E. Havel, D. L. Helmers, J. B. Hooker, J. R. Jones, M. F. Knowlton, J. Kubisiak, J. Mazourek, A. C. McColpin, R. B. Renken, and R. D. Semlitsch. 1998. Flooding to restore connectivity of regulated, large- river wetlands. BioSciene 48:721-733. Hammen, J. J., E. Pherigo, W. Doyle, J. Finley, K. Drews, and J. M. Goeckler. 2019. A comparison between conventional boat electrofishing and the electrified dozer trawl for capturing Silver Carp in tributaries of the Missouri River, Missouri. North American Journal of Fisheries Management 39:582-588. Haupt, K. J., and Q. E. Phelps. 2016. Mesohabitat association in the Mississippi River Basin: a long-term study on the catch rates and physical habitat association of juvenile Silver Carp and two native planktivores. Aquatic Invasions 11:93-99. Hayer, C.-A., J. J. Breeggemann, R. A. Klumb, B. D. S. Graeb, and K. N. Bertrand. 2014. Population characteristics of Bighead and Silver Carp on the Northwestern front of their North American invasion. Aquatic Invasions 9:289-303. Hüssy, K., K. E. Limberg, H. de Pontual, O. R. B. Thomas, P. K. Cook, Y. Heimbrand, M. Blass, and A. M. Sturrock. Trace element patterns in otoliths: the role of biomineralization. Reviews in Fisheries Science and Aquaculture. Hosen, J. D., K. S. Aho, A. P. Appling, E. C. Creech, J. H. Fair, R. O. Hall Jr. E. D. Kyzivat, R. S. Lowenthal, S. Matt, J. Morrison, J. E. Saiers, J. B. Shanley, L. C. Weber, B. Yoon, and P. A. Raymond. 2019. Enhancement of primary production during drought in a temperate watershed is greater in large rivers than headwater streams. Limnology and Oceanography 64:1458-1472. Kolar, C. S., D. C. Chapman, W. R. Courtenay Jr., C. M. Housel, and J. D. Williams. 2005. Asian carps of the Genus Hypophthalmichthys (Pisces, Cyprinidae) – A biological synopsis and environmental risk assessment. National Invasive Species Council materials. 5. Love, S. A., N. J. Lederman, T. Widloe, and G. W. Whitledge. 2019. Sources of Bighead Carp and Silver Carp found in Chicago urban fishing program ponds. Transactions of the American Fisheries Society 148:417-425. 99

Lu, G., C. Wang, J. Zhao, X. Liao, J. Wang, M. Luo, L Zhu, L. Bernatzhez, and S. Li. 2020. Evolution and genetics of Bighead and Silver Carps: Native population conservation versus species control. Evolutionary Applications 13:1351- 1362.Pegg Makinster, A. S., and C. P. Paukert. 2008. Effects and utility of minimum length limits and mortality caps for Flathead Catfish in discrete reaches of a large prairie river. North American Journal of Fisheries Management 28:97-108. Melancon, S., B. J. Fryer, S. A. Ludsin, J. E. Gagnon, and Z. Yang. 2005. Effects of crystal structure on the uptake of metals by Lake Trout (Salvelinus namaycush) otoliths. Canadian Journal of Fisheries and Aquatic Sciences 62:2609-2619. Norman, J. D., and G. W. Whitledge. 2015. Recruitment sources of invasive bighead carp (Hypophthalmichthys nobilis) and silver carp (H. molitrix) inhabiting the Illinois River. Biological Invasions 17:2999-3014. Pegg, M. A., and J. H. Chick. 2010. Habitat Improvement in Altered Systems. Pages 295- 324 in W. A. Hubert and M. C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Pegg, M. A., C. L. Pierce, and A. Roy. 2003. Hydrological alteration along the Missouri River Basin: A time series approach. Aquatic Sciences 65:63-72. Perkin, J. S., Z. R. Shattuck, P. T. Bean, and T. H. Bonner. 2010. Movement and microhabitat association of Guadalupe Bass in two Texas river. North American Journal of Fisheries Management 30:33-46. Peters, L. M., M. A. Pegg, and U. L. Reinhardt. 2006. Movement of adult radio-tagged Bighead Carp in the Illinois River. Transactions of the American Fisheries Society 135:1205-1212. Phelps, Q. E., G. W. Whitledge, S. J. Tripp, K. T. Smith, J. E. Garvey, D. P. Herzog, D. E. Ostendorf, J. W. Ridings, J. W. Crites, R. A. Hrabik, W. J. Doyle, and T. D. Hill. 2012. Identifying river of origin for age-0 Scaphirhynchus sturgeons in the Missouri and Mississippi rivers using fin ray microchemistry. Canadian Journal of Fisheries and Aquatic Sciences 69:930-941. Porreca, A. P., W. D. Hintz, G. W. Whitledge, N. P. Rude, E. J. Heist, and J. E. Garvey. 2016. Establishing ecologically relevant management boundaries: linking movement ecology with the conservation of Scaphirhynchus sturgeon. Canadian Journal of Fisheries and Aquatic Sciences 73:877-884. Pracheil, B. M., R. George, and B. C. Chakoumakos. 2019. Significance of otolith calcium carbonate crystal structure diversity to microchemistry studies. Review in Fish Biologies and Fisheries 29:569-588. 100

Prechtel, A. R., A. A. Coulter, L. Etchison, P. R. Jackson, and R. R. Goforth. 2018. Rang estimates and habitat use of invasive Silver Carp (Hypophthalmichthys molitrix): evidence of sedentary and mobile individuals. Hydrobiologia 805:203-218. Quist, M. C., and C. S. Guy. 1999. Spatial variation in population characteristics of the Shovelnose Sturgeon in the Kansas River. The Prairie Naturalist 31:65-74. Quist, M. C., J. S. Tillma, M. N. Burlingame, and C. S. Guy. 1999. Overwinter habitat use of shovelnose sturgeon in the Kansas River. Transactions of the American Fisheries Society 128:522-527. R Core Team. 2020. R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Rodríguez, M. A. 2002. Restricted movement in stream fish: the paradigm is incomplete, not lost. Ecology 83:1-13. Spurgeon, J. J., M. A. Pegg, and N. M. Halden. 2018. Mixed-origins of channel catfish in a large-river tributary. Fisheries Research. 198:195-202. Steffensen, K. D. and G. E. Mestl. 2016. Assessment of Pallid Sturgeon relative condition in the upper channelized Missouri River. Journal of Freshwater Ecology 31:583- 595. Sturrock, A. M., E. Hunter, A. Milton, EIMF, R. C. Johnson, C. P. Waring, and C. N. Trueman. 2015. Quantifying physiological influences on otolith microchemistry. Methods in Ecology and Evolution 6:806-816. Therneau, T. M., E. J. Atkinson, and Mayo Foundation. 2019. An introduction to recursive partitioning using the RPART routines. R Package Version 4.1-15 pages 1-14. Thomas, O. R. B., K. Ganio, B. R. Roberts, and S. E. Swearer. 2017. Trace element- protein interactions in the enolymph of the inner ear of fish: implications for environmental reconstruction using otolith chemistry. Metallomics 9:239-249. Tsehaye, I., M. Catalano, G. Sass, D. Glover, and B. Roth. 2013. Prospects for fishery- induced collapse of invasive Asian carp in the Illinois River. Fisheries 38:445- 454. Whitledge, G. W., B. Knights, J. Vallazza, J. Larson, M. J. Weber, J. T. Lamer, Q. E. Phelps, and J. D. Norman. 2018. Identification of Bighead Carp and Silver Carp early-life environments and inferring Lock and Dam 19 passage in the upper Mississippi River: insights from otolith chemistry. Biological Invasions. Whitledge, G. W., B. M. Johnson, P. J. Martinez, and A. M. Martinez. 2007. Sources of nonnative centrarchids in the upper Colorado River revealed by stable isotope and microchemical analyses of otoliths. Transactions of the American Fisheries Society 136: 1263-1275. 101

Zeigler, J. M. and G. W. Whitledge. 2010. Assessment of otolith chemistry for identifying source environment of fishes in the lower Illinois River, Illinois. Hydrobiologia 638:109-119. 102

Table 4-1: Classification matrix for Silver Carp collected from Segment 2 of the Kansas

River and from the Wakarusa River in 2018, and from isolated backwaters of the

Missouri River used to build and test the recursive partitioning tree.

Sampled

Predicted Kansas River Missouri River Wakarusa River Kansas River 12 2 1 Missouri River 1 2 0 Wakarusa River 0 0 0

103

Table 4-2: Percent transient adult and juvenile Silver Carp in Segment 1 and Segment 2 of the Kansas River, and in both segments combined in 2018 and 2019. Total number of fish captured are in parenthesis.

Segments Segment 1 Segment 2 Combined Adult Silver Carp 2018 49% (n = 69) 49% (n = 72) 49% (n = 141) 2019 22% (n = 18) 45% (n = 80) 41% (n = 98) Years Combined 44% (n = 87) 47% (n = 152) Juvenile Silver

Carp 2018 N/A (n = 0) N/A (n = 0) N/A (n = 0) 2019 34% (n = 32) 40% (n = 5) 35% (n = 37) Years Combined 34% (n = 32) 40% (n = 5)

104

Table 4-3: Number of transient adult and juvenile Silver Carp captured in the Kansas

River that exhibit 1, 2, and 3, movement events into the Missouri River for each segment.

Natal origins from the Missouri River is the number of transient individuals with

Missouri River Sr:Ca signatures at the nucleus of the otolith.

Number of Movement Events Natal Origins from the 1 2 3 Missouri River Adult Silver Carp Segment 1 25 12 1 10 Segment 2 57 12 2 9 Juvenile Silver Carp Segment 1 9 2 0 5 Segment 2 1 1 0 1

105

Figure 4-1: Kansas River study area. Segment 1 (27 rkm) is between the confluence with the Missouri River and the Johnson County Weir, Segment 2 (56 rkm) is between the

Johnson County Weir and Bowersock Dam, and Segment 3 (58 rkm) is between

Bowersock Dam and the Topeka Weir. Red lines indicate the three main barriers on the river and open circles indicate water chemistry sample collection sites. 106

Figure 4-2: Comparison of water microchemistry signatures from the Kansas, Missouri, and Wakarusa Rivers. The horizontal solid line within the box represents the median value, upper and lower limits of the box is quartile ranges, and whiskers are 95% confidence intervals of the median. Points represent data outside of the 95% confidence interval. Median water Sr:Ca and Ba:Ca ratios do not differ between rivers that bear the same letter above the box plot.

107

Figure 4-3: Comparison of otolith microchemistry signatures (~ 30µm of data isolated from the edge of the otolith) from Silver Carp captured from Segment 2 of the Kansas

River in 2018, the Wakarusa River in 2018, and isolated backwaters of the Missouri

River. The horizontal solid line within the box represents the median value, upper and lower limits of the box are quartile ranges, and whiskers are 95% confidence intervals of the median. Points represent data outside of the 95% confidence interval. Median otolith

Sr:Ca and Ba:Ca ratios do not differ between rivers that bear the same letter above the box plot.

108

Figure 4-4: Recursive partitioning tree used to classify environmental history of Silver

Carp between the Kansas River and the Missouri River. The tree was pruned at size = 2 with a complexity parameter (cp) of 0.14.

109

Figure 4-5: Example of ablation data from three Silver Carp captured from the Kansas

River that exhibit trace elemental signatures indicative of the Missouri River. The horizontal line represents the threshold between the Kansas and Missouri Rivers (Sr:Ca =

2,082 µmol/mol) where signatures above the line are from the Missouri River. The shaded region is data that corresponds to the nucleus of the otolith (i.e., natal origins).

Fish number 180208-2 had natal origin signatures from the Missouri River as well as another movement event with signatures from the Missouri River later in life. Fish

180236-18 and 190241-8 both have movement events with signatures from the Missouri

River. 110

Figure 4-6: Percent of laser ablation points above 2,082 µmol/mol Sr:Ca threshold for each individual transient fish by total length. Data points greater than 2,082 µmol/mol

Sr:Ca were indicative of Missouri River trace element signatures in lapilli otoliths of

Silver Carp. 111

CHAPTER 5: MANAGEMENT AND RESEARCH RECOMMENDATIONS

Silver Carp exhibited spatiotemporal differences in population dynamics and vital rates across their distribution. Silver Carp captured in Segment 2 were longer in length, had a greater length-at-age, and occurred at lower relative abundances compared to fish captured in Segment 1. Additionally, juvenile Silver Carp were rare in Segment 2

(observed in this segment only in 2019), but were abundant in Segment 1 in both sampling years. These results suggests the Johnson County Weir functions as a partial barrier to movement. Additionally, the lack of juveniles in Segment 2 suggests reproduction may be limited in this segment (Chapter 2).

Our results from the otolith microchemistry analysis suggest the population of

Silver Carp in the Kansas River is comprised predominantly of residential individuals.

Additionally, our results indicate the bulk of the recruitment occurs within the Kansas

River itself with few individuals exhibiting natal origin signatures of the Missouri River.

Transient Silver Carp displayed signatures indicative of the Missouri River for relatively short durations, with multiple movement events observed in some individuals. These results suggest occupancy in the Missouri River is brief and Silver Carp may use the main-stem Missouri River more as a movement corridor as they move between habitats adjacent to the main-stem river (Chapter 4).

We did not detect Silver Carp or Bighead Carp in Segment 3 with conventional gears (Chapter 2) or with the environmental DNA (eDNA) assay. These results suggest

Asian carp are not present in reaches upstream of Bowersock Dam in substantial numbers 112 and that Bowersock Dam is able to impede immigration of Silver Carp to upstream reaches. Black Carp were detected near the Kansas-Missouri confluence using the eDNA assay, marking the northernmost detection of this species in the Missouri River (Chapter

3).

Management Recommendations

Management of invasive species often focuses on eradication and population control. Eradication in large river systems – like the Kansas River – is nearly impossible after a species has become established, and management goals should shift to population control. Additionally, mitigating secondary introductions is crucial for ensuring the resiliency of ecosystems in non-infested reaches (Kolar et al. 2010; Vander Zanden and

Olden 2008). Therefore, management strategies for Asian carp in the Kansas River should focus on population control, mitigating secondary introductions, and continued monitoring in non-infested reaches.

Population Control

The Kansas River affords a unique opportunity for population control and possible population reduction of Asian carp given the characteristics of this population.

The results of our otolith microchemistry analysis indicate that the bulk of recruitment of

Silver Carp occurs within the Kansas River (Chapter 4). Segment 1 is likely of particular importance for reproduction as age-0 and juvenile Silver Carp were abundant in Segment

1 but rare in Segment 2 (Chapter 2). Therefore, we recommend that population control strategies are focused on Segment 1. Here, we explore possible management strategies for population control. 113

 Mechanical removal – Mechanical removal efforts would likely be most effective in

the fall or late summer. For example, Ridgeway et al. (2020) found that catch per unit

effort (CPUE) was the highest during the fall in Lake Barkley, Lake Kentucky and

backwaters of the Illinois River. Additionally, removal efforts during this time could

target a larger proportion of transient individuals because movement distances are

reduced during low flow periods (DeGrandchamp et al. 2008; Coulter et al. 2016).

Segment 2 may be a population sink for Silver Carp, maintained by emigration

through or from Segment 1. Therefore, removal efforts focused on Segment 1 would

likely have trickle-down effects in Segment 2. Specialized gears such as the

electrified dozer trawl (e.g., Hammen et al 2019) or electrified paupier (e.g.,

Ridgeway et al. 2020) would be optimal for removal of Silver Carp because these

gears have higher catch rates than conventional electrofishers (Chapter 2; Hammen et

al. 2019). Additionally, the specialized gears sample a wider breadth of size groups

than conventional gears (Chapter 2). When specialized gears are not available, high

frequency electrofishing should be used (Chapter 2), employing methods described

by Bouska et al. (2017). Recent research into diel sampling patterns indicate that

night sampling (e.g., first 5 hours after sunset) has the highest CPUE for Silver Carp

(Ridgeway et al. 2020). However, night electrofishing would only be safe during

periods of low flows such as those in 2018 (e.g. Chapter 2) and standardization across

years would be difficult if flows did not allow for safe operation of gears at night.

 Chemical treatments – We observed high concentrations of Silver Carp in most

backwater or slack water habitats adjacent to the main channel in Segments 1 and 2 114

during the high water events in 2019. Many of these backwater habitats were

relatively shallow compared to the main channel habitat and proved difficult to

maneuver boat-mounted electrofishers in for mechanical sampling. As such, these

areas could be isolated with a net barrier and targeted chemical treatments could be

applied for the removal of large numbers of Silver Carp and Bighead Carp. Rotenone

should be used in chemical treatments for Silver Carp and Bighead Carp as these

fishes are far more sensitive to rotenone than antimycin a (Chapman et al. 2003).

 Piscivory – Robust populations of Blue Catfish Ictalurus furcatus and Flathead

Catfish Plyodictis olivaris are present within the Kansas River, with some individuals

reaching large enough sizes to forage on Silver Carp and Bighead Carp (Paukert and

Makinster 2009; Dean 2020). Furthermore, piscivory of Silver Carp by both catfish

species has been documented in this system (Dean, unpublished data). Managing the

Kansas River as a trophy fishery for these catfish species could promote increased

piscivory on Asian carp.

 Trojan Y Chromosome – This strategy aims to change the sex ratio of the population

to contain a higher proportion of male individuals over time and lead to the eventual

eradication of the target species (Teem and Guitierrez 2014). However, this strategy

would prove difficult in large, connected river systems for mobile species because the

number of introduced specimens needed for success would be far too great.

Monitoring for Asian Carp in Segment 3

Asian carp are notoriously difficult to detect when they are at low population densities. Our sampling approach (both mechanical sampling and eDNA sampling) failed 115 to detect Silver Carp or Bighead Carp upstream of Bowersock Dam (Chapter 2; Chapter

3); however, we recommend continued monitoring in this segment. A combination of eDNA techniques and mechanical sampling would be best as neither technique is infallible.

 Environmental DNA – Our results indicate that eDNA techniques require further

refinement for applications in large, turbid systems (Chapter 3). Centrifugation of

turbid samples may be the best means to process water samples from the Kansas

River. Additionally, larger sample sizes – either more samples or a larger volume per

sample – is likely needed to detect Asian carp at low densities (Willoughby et al.

2016; Sepulveda et al. 2019). Using PCR methods that are more sensitive at low

concentrations of target aqueous DNA (i.e., quantitative PCR or digital droplet PCR)

could improve detectability as well (Nathan et al. 2014; Wilcox et al. 2016; Xia et al.

2018).

 Mechanical sampling – Conventional gears would be best used in targeted sampling,

where effort is focused in preferred habitat types of Silver Carp and Bighead Carp in

Segment 3 (e.g., slow moving backwaters and barrier effluents), and Black Carp in

Segment 1(e.g., deep, slow moving pools). Targeted sampling could also be

performed in areas where any target species are detected with eDNA assays. For

example, sampling for Black Carp in the Kansas River should be focused near the

confluence with the Missouri River because this is where we detected them with our

eDNA assay (Chapter 3). Additionally, this area of the river has deep pools and slow 116

currents that Black Carp have shown a preference for in their native range (Nico et al.

2005).

Movement Deterrents

Secondary introductions into Segment 3 is most probable during high water events, such as those in 2019. For example, we observed multiple individual Silver Carp attempting to traverse Bowersock Dam during the largest discharge event in 2019 (e.g., approximately 2,800 m3/second; Chapter 2). Additional measures impeding movement over Bowersock Dam may be considered to mitigate upstream infestation risks. Here, we explore possible strategies for deterrents and mitigating secondary introductions.

 Acoustic deterrents – Acoustic deterrents have shown potential in limiting Asian carp

movement (Taylor et al. 2005; Murchy et al. 2017; Vetter et al. 2017a; Vetter et al.

2017b). This strategy could be implemented as an additional deterrent measure on

Bowersock Dam. For example, attaching underwater speakers and turning them on

only during high water events could push fish downstream of the barrier (Murchy et

al. 2017) and reduce attempts to traverse the dam. Attaching the speakers directly

below the powerhouse spillway located near the north shore would likely be the most

effective because this area was where Silver Carp were attempting to traverse the

dam. This deterrent strategy could also be implemented on or near the Johnson

County Weir to reduce immigration rates from Segment 1 into Segment 2.

 SPA driven BAFF – Sound Projector Array driven BioAcoustic Fish Fence system

(SPA driven BAFF; Fish Guidance Systems, Ltd, UK) described in Taylor et al.

(2005), or a similar acoustic fence configuration, near the confluence with the 117

Missouri River could limit immigration of transient individuals into the Kansas River.

For example, a strategy much like this was implemented in the Laurentian Great

Lakes to block passage for Sea Lamprey’s Petromyzon marinus into tributaries for

spawning (Lavis et al. 2003).

 Regulating discharge – Discharge in the Kansas River drainage is controlled by 18

federal reservoirs and over 13,000 smaller impoundments (Quist et al. 1999;

Makinster and Paukert 2008). Controlled and coordinated discharge throughout the

Kansas River drainage could be used to manipulate flows that are not conducive for

certain life-history traits of Silver Carp. For example, limiting flow in the spring

when Silver Carp movement rates are the highest could reduce immigration from

Segment 1 into Segment 2. Additionally, managing flows to maintain a velocity

greater than 1.6 m/second could severely inhibit spawning activity as well as the

survival of newly hatched Silver Carp (Chen et al. 2020).

 Public outreach – Currently, the Kansas Department of Wildlife, Parks and Tourism

(KDWPT) has an outreach program educating recreators about the deleterious

consequences of moving Asian carp. Maintaining this program and focusing on

educating the public about the negative impacts Asian carp have on native fishes and

recreational boating is imperative.

Future Research Needs

1. Population estimates in Segments 1 and 2 – Estimating population size will provide

insight as to the target number of individuals to remove to decrease population size of

Silver Carp and Bighead Carp. Additionally, tracking estimated population sizes 118

through time would provide feedback if population control measures are successful.

A robust design would likely be the model of best fit for population size estimates in

Segments 1 and 2.

2. Tracking movements in the Kansas River – These data will provide managers insight

as to where removal efforts would likely be most effective. For example, tracking fish

through the spring could reveal spawning aggregate locations, or identify areas fishes

congregate in the late summer and early autumn. Additionally, passive transponders

located near the Johnson County Weir and the confluence with the Missouri River

will provide insight into movement rates between Segments 1 and 2 and with the

Missouri River. These data could also identify flows that facilitate movement

between Segments 1 and 2.

3. Main-stem Missouri River occupancy durations – Results from our microchemistry

analysis suggests transient Silver Carp captured in the Kansas River occupied the

Missouri River for brief periods (Chapter 4). These findings need to be further

explored for individuals captured throughout the main stem Missouri River. These

data will provide insight as to where management efforts need to be focused

throughout the basin.

4. Water-otolith trace elemental relationships – Our otolith microchemistry results

indicate that the relationship between water and otolith trace elemental signatures

may be logistic rather than linear. The role physiological processes governing trace

element uptake and transport have in water-otolith trace element relationships is 119

relatively unknown. Further research in these relationships is warranted for more

accurate interpretations of the results of this technique.

5. Sediment eDNA sampling in turbid systems – Turner et al. (2015) found DNA

extracted from sediment samples occurred at higher concentrations than DNA

extracted from water samples. Additionally, the DNA found in the sediment persisted

far longer than DNA suspended in the water column (Turner et al. 2015; Buxton et al.

2018). Sampling the sediment in turbid rivers could prove to be more efficient and

sensitive means for detecting Asian carp than surface water samples.

6. Standardized eDNA sample sizes – Multiple factors influence detectability of aqueous

DNA during eDNA assays, including the volume of water in the system at the time of

sample collection. Investigating what volume of water needs to be sampled compared

to the volume of water in either lentic or lotic systems is vital for confidence in eDNA

results and for standardization of this technique.

7. Black Carp presence in the Kansas River – We had limited evidence suggesting

Black Carp are present in the Kansas River. Further eDNA sampling could help

confirm our results and determine if mechanical sampling efforts are warranted in the

Kansas River.

8. Fish assemblage structure in Segments 2 and 3 – Segments 2 and 3 have similar

habitat types and riparian influences (Paukert and Makinster 2009). As such,

differences in native fish and gamefish assemblages between Segments 2 and 3 will

provide insight into the impacts establishment of Asian carp may have on the fish

assemblages in Segment 3 as well as in other Great Plains rivers. 120

Conclusions

Population control of Asian carp in the Kansas River will likely need a multi- faceted approach to successfully manage and reduce the population within the Kansas

River. Similarly, multiple techniques should be used for monitoring protocols in reaches upstream of Bowersock Dam. Continual management actions will be required due to the unimpeded connectivity with the Missouri River and the potential for reproduction within the Kansas River itself. A basin-wide pest management plan adhered to by the multiple jurisdictions throughout the Missouri River basin is vital. 121

Literature Cited Bouska, W. W., D. C. Glover, K. L. Bouska, and J. E. Garvey. 2017. A refined electrofishing technique for collecting Silver Carp: implications for management. North American Journal of Fisheries Management 37:101-107. Buxton, A. S., J. J. Groombridge, and R. A. Griffin. Seasonal variation in environmental DNA detection in sediment and water samples. PLoS ONE e0191737. Chapman, D., J. Fairchild, B. Carollo, J. Deters, K. Feltz, and C. Witte. 2003. An examination of the sensitivity of Bighead Carp and Silver Carp to antimycin a and rotenone. Final report to the Asian carp Rapid Response Committee of the Dispersal Barrier Advisory Panel to the Great Lakes on Aquatic Nuisance Species. U.S. Geological Survey, Columbia Environmental Research Center, Columbia, Missouri. Chen, Q., J. Zhang, Y. Chen, K. Mo, J. Wang, L. Tang, Y. Lin, L. Chen, Y. Gao, W. Jiang, and Y. Zhang. Inducing flow velocities to manage fish reproduction in regulated rivers. Engineering. Coulter, A. A., E. J. Bailey, D. Keller, and R. R. Goforth. 2016. Invasive Silver Carp movement patterns in the predominantly free-flowing Wabash River (Indiana, USA). Biological Invasions 18:471-485. Dean, Q. J. 2020. Population characteristics and movement of Blue Catfish in the Kansas River. Master’s thesis. University of Nebraska, Lincoln. DeGrandchamp, K. L., J. E. Garvey, and R. E. Colombo. 2008. Movement and habitat selection by invasive Asian carp in a large river. Transactions of the American Fisheries Society 137:45-56. Hammen, J. J., E. Pherigo, W. Doyle, J. Finley, K. Drews, and J. M. Goeckler. 2019. A comparison between conventional boat electrofishing and the electrified dozer trawl for capturing Silver Carp in tributaries of the Missouri River, Missouri. North American Journal of Fisheries Management 39:582-588. Kolar, C. S., W. R. Courtenay, and L. G. Nico. 2010. Managing undesired and invading fishes. Pages 213-259 in W.A. Hubert and M.C. Quist, editors. Inland fisheries management in North America, 3rd edition. American Fisheries Society, Bethesda, Maryland. Lavis, D. S., A. Hallett, E. M. Koon, and T. C. McAuley. 2003. History and advances in barriers as an alternative method to suppress Sea Lampreys in the Great Lakes. Journal of Great Lakes Research 29(Supplement 1):362-372. Makinster, A. S., and C. P. Paukert. 2008. Effects and utility of minimum length limits and mortality caps for Flathead Catfish in discrete reaches of a large prairie river. North American Journal of Fisheries Management 28:97-108.Fisheries Management and Ecology 24:208-216. 122

Murchy, K. A., A. R. Cupp, J. J. Amberg, B. J. Vetter, K. T. Fredricks, M. P. Gaikowski, and A. F. Mensinger. 2017. Potential implications of acoustic stimuli as a non- physical barrier to Silver Carp and Bighead Carp. Nathan, L. M., M. Simmons, B. J. Wegleitner, C. L. Jerde, and A. R. Mahon. 2014. Quantifying environmental DNA signals for aquatic invasive species across multiple detection platforms. Environmental Science and Technology 48:12800- 12806. Nico, L. G., J. D. Williams, and H. L. Jelks. 2005. Black Carp: biological synopsis and risk assessment of an introduced fish. American Fisheries Society, Special Publication 32, Bethesda, Maryland. Paukert, C. P., and A. S. Makinster. 2009. Longitudinal patterns in Flathead Catfish relative abundance and length at age within a large river: effects of an urban gradient. River Research and Applications 25:861-873. Quist, M. C., J. S. Tillma, M. N. Burlingame, and C. S. Guy. 1999. Overwinter habitat use of shovelnose sturgeon in the Kansas River. Transactions of the American Fisheries Society 128:522-527. Ridgeway, J. L., J. M. Goeckler, J. Morris, J. J. Hammen. 2020. Diel influences on Silver Carp catch rates using an electrified paupier in lentic habitats. North American Journal of Fisheries Management 40:1023-1031. Sepulveda, A. J., J. Schabacker, S. Smith, R. Al-Chokhachy, G. Luikart, and S. J. Amish. 2019. Improved detection of rare, endangered and invasive trout in using a new large-volume sampling method for eDNA capture. Environmental DNA 1:227- 237. Taylor, R. M., M. A. Pegg, and J. H. Chick. 2005. Response of Bighead Carp to a bioacoustics behavioral fish guidance system. Fisheries Management and Ecology 12:283-286. Teem, J. L., and J. B. Gutierrez. 2014. Combining the Trojan Y chromosome and daughterless carp eradication strategies. Biological Invasions 16:1231-1240. Turner, C. R., K. L. Uy, and R. C. Everhart. 2015. Fish environmental DNA is more concentrated in aquatic sediments that surface water. Biological Conservation 183:93-102. Vander Zanden, J. M. and J. D. Olden. 2008. A management framework for preventing the secondary spread of aquatic invasive species. Canadian Journal of Fisheries and Aquatic Sciences 65:1512-1522. 123

Vetter, B. J., A. R. Cupp, K. T. Fredricks, M. P. Giakowski, and A. F. Mensinger. 2017a. Acoustic deterrence of Silver Carp (Hypophthalmichthys molitrix). Biological Invasions 17:3383-3392. Vetter, B. J., K. A. Murchy, A. R. Cupp, J. J. Amberg, M. P. Giakowski, and A. F. Mansinger. 2017b. Acoustic deterrence of Bighead Carp (Hypophthalmichthys nobilis) to a broadband sound stimulus. Journal of Great Lakes Research 43:163:171. Wilcox, T. M., K. S. McKelvey, M. K. Young, A. J. Sepulveda, B. B. Shepard, S. F. Jane, A. R. Whiteley, W. H. Lowe, and M. K. Schwartz. 2016. Understanding environmental DNA detection probabilities: a case study using stream-dwelling char Salvelinus fontinalis. Biological Conservation 194:209-216. Willoughby, J. R., B. K. Wijayawardena, M. Sundaram, R. K. Swihart, and J. A. Dewoody. 2016. The importance of including imperfect detection models in eDNA experimental design. Molecular Ecology 16:837-844. Xia, Z., M. L. Johansson, Y. Gao, L. Zhang, G. D. Haffner, H, J, MacIsaac, and A. Zhan. 2018. Conventional versus real-time quantitative PCR for rare species detection. Ecology and Evolution 8:11799-11807.

124

APPENDIX A

Supplemental tables and figures pertaining to Chapter 2

125

Table A-1: Total counts of all fishes (common names) captured in Segments 1, 2, and 3 of the Kansas River with the High Frequency Electrofishing method over all sampling years.

Species Segment 1 Segment 2 Segment 3 Shovelnose Sturgeon 2 0 1 Longnose Gar 31 28 6 Shortnose Gar 4 12 6 Goldeye 17 8 6 Gizzard Shad 58 32 65 Red Shiner 177 361 422 Golden Shiner 0 1 1 Emerald Shiner 6 23 21 Sand Shiner 5 18 22 Bluntnose Minnow 3 13 20 Bullhead Minnow 12 40 40 Grass Carp 5 3 1 Common Carp 6 12 10 Silver Carp 215 280 0 Bighead Carp 8 6 0 River Carpsucker 32 118 35 Quillback 0 0 3 Blue Sucker 0 15 0 Smallmouth Buffalo 18 41 49 Bigmouth Buffalo 2 6 7 Blue Catfish 6 6 11 Channel Catfish 1 13 8 Flathead Catfish 1 16 13 126

Table A-1: continued

Species Segment 1 Segment 2 Segment 3 Western Mosquitofish 0 0 1 Wiper 13 5 5 Lepomis Spp. 3 0 0 Green Sunfish 0 1 0 Orangespotted Sunfish 3 0 4 Bluegill 6 4 5 Longear Sunfish 0 4 1 Micropterous Spp. 1 1 5 Smallmouth Bass 0 0 1 Largemouth Bass 0 1 3 White Crappie 7 7 2 Black Crappie 0 3 1 Ozark Logperch 0 3 0 Walleye 0 1 0 Freshwater Drum 17 17 12

127

Table A-2: Total counts of all fishes (common names) captured in Segments 1, 2, and 3 of the Kansas River with the Low Frequency Electrofishing method over all sampling years.

Species Segment 1 Segment 2 Segment 3 Shovelnose Sturgeon 18 5 1 Longnose Gar 16 6 6 Shortnose Gar 9 1 1 Goldeye 2 16 3 Gizzard Shad 7 5 10 Red Shiner 33 55 45 Emerald Shiner 0 13 0 Bluntnose Minnow 0 0 1 Bullhead Minnow 0 27 6 Grass Carp 0 2 1 Common Carp 0 9 4 Silver Carp 404 152 0 Bighead Carp 11 12 0 River Carpsucker 25 28 26 Quillback 0 1 0 Blue Sucker 2 16 1 Smallmouth Buffalo 33 13 14 Bigmouth Buffalo 1 2 3 Blue Catfish 335 369 524 Channel Catfish 44 212 135 Flathead Catfish 178 318 231 Western Mosquitofish 0 25 0 Wiper 0 0 1 128

Table A-2: Continued

Species Segment 1 Segment 2 Segment 3 Centrarchidae Spp. 0 0 1 Bluegill 1 3 0 Largemouth Bass 0 2 0 White Carppie 2 0 0 Black Crappie 0 0 1 Sauger 0 1 0 Freshwater Drum 4 6 8

129

Table A-3: Total counts of all fishes (common names) captured in Segments 1, 2, and 3 of the Kansas River with the Electrified Dozer Trawl over all sampling years.

Species Segment 1 Segment 2 Segment 3 Shovelnose Sturgeon 0 4 0 Longnose Gar 13 16 4 Shortnose Gar 7 2 7 Goldeye 36 2 3 Gizzard Shad 268 89 119 Red Shiner 86 142 59 Emerald Shiner 114 11 9 Suckermouth Minnow 1 0 0 Bluntnose Minnow 0 18 0 Grass Carp 4 0 0 Common Carp 1 1 0 Silver Carp 347 39 0 Bighead Carp 0 1 0 River Carpsucker 2 4 18 Blue Sucker 0 1 0 Smallmouth Buffalo 2 2 4 Bigmouth Buffalo 1 1 3 Blue Catfish 7 10 9 Channel Catfish 0 12 41 Flathead Catfish 0 1 0 Western Mosquitofish 0 0 2 Wiper 17 5 10

130

Table A-3: Continued

Species Segment 1 Segment 2 Segment 3 Orangespotted Sunfish 2 1 3 Bluegill 20 21 15 Micropterous Spp. 1 0 0 Largemouth Bass 2 0 1 White Crappie 52 7 5 Freshwater Drum 10 16 11

131

Table A-4: Total counts of all fishes (common names) captured in Segments 1, 2, and 3 of the Kansas River with the mini-fyke nets over all sampling years.

Species Segment 1 Segment 2 Segment 3 Longnose Gar 10 15 2 Shortnose Gar 11 8 8 Gizzard Shad 70 254 126 Central Stoneroller 0 0 9 Speckled Chub 0 5 0 Creek Chub 6 0 0 Red Shiner 180 509 152 Notropis Spp. 59 0 0 Emerald Shiner 5 101 19 Sand Shiner 11 73 59 Suckermouth Minnow 18 2 2 Bluntnose Minnow 7 36 2 Bullhead Minnow 206 477 105 Common Carp 3 5 2 Silver Carp 13 2 0 Bighead Carp 1 0 0 River Carpsucker 13 114 32 Quillback 7 17 0 Smallmouth Buffalo 3 46 11 Bigmouth Buffalo 21 1 9 Redhorse 0 1 0 Yellow Bullhead 11 1 1 Blue Catfish 5 6 1 Channel Catfish 7 130 11

132

Table A-4: Continued

Species Segment 1 Segment 2 Segment 3 Flathead Catfish 4 5 0 Western Mosquitofish 13 48 2 Wiper 20 2 20 Green Sunfish 0 5 0 Orangespotted Sunfish 13 15 0 Bluegill 53 23 4 Micropterous Spp. 12 140 52 Largemouth Bass 9 1 4 White Crappie 103 113 47 Pomoxis Spp. 0 20 0 Orangethroated Darter 1 0 2 Ozark Logperch 3 5 3 Freshwater Drum 103 88 40

133

Table A-5: Total counts of all fishes (common names) captured in Segments 1, 2, and 3 of the Kansas River with the hoop nets over all sampling years.

Species Segment 1 Segment 2 Segment 3 Shovelnose Sturgeon 0 5 1 Smallmouth Buffalo 0 1 1 Channel Catfish 0 1 0 Flathead Catfish 0 1 2 Orangespotted Sunfish 0 0 1 White Crappie 0 0 16

134

Figure A-1: Mean relative weights of male and female Silver Carp captured out of

Segments 1 and 12of the Kansas River. Error bars represent the 95% confidence intervals around the mean. Grey points are the mean relative weight of fish captured in 2018 and the black points are the mean relative weight of fish captured in 2019.

135

Figure A-2: Predicted lengths-at-age from Von-Bertalanffy growth models for Silver

Carp populations in the Illinois River (red circles and solid line), Segment 1 (light gray squares and dashed line), Segment 2 (dark gray crosses and dashed line), Wabash River

(blue stars and dashed line), Middle Mississippi River (green triangles and dotted line), and South Dakota tributaries (yellow crosses inside boxes and dotted line) (Williamson and Garvey 2005; Hayer et al. 2014; Stuck et al. 2015).

136

APPENDIX B

Supplemental tables and figures pertaining to Chapter 3

137

Table B-1: Results from the environmental DNA analysis for Silver Carp from each sample collected at each sampling site in the Kansas River. Positive detections are denoted as (+) and negative detections as (-). Sites P and 1-3 were in Segment 1, sites 4-8 were in Segment 2, and sites 9-13 were in Segment 3. Samples 1 and 2 were collected near the South bank, samples 3 and 4 were collected from the thalweg, and samples 5 and

6 were collected from the North bank.

Sample Number Site 1 2 3 4 5 6 P + + + NA NA NA

Segment 1 ------1 2 ------3 ------4 ------5 ------Segment 6 ------2 7 ------8 ------9 ------10 ------Segment 11 ------3 12 ------13 ------

138

Table B-2: Results from the environmental DNA analysis for Bighead Carp from each sample collected at each sampling site in the Kansas River. Positive detections are denoted as (+) and negative detections as (-). Sites P and 1-3 were in Segment 1, sites 4-8 were in Segment 2, and sites 9-13 were in Segment 3. Samples 1 and 2 were collected near the South bank, samples 3 and 4 were collected from the thalweg, and samples 5 and

6 were collected from the North bank.

Sample Number Site 1 2 3 4 5 6 P - - - NA NA NA

Segment 1 ------1 2 ------3 ------4 ------5 ------Segment 6 ------2 7 ------8 ------9 ------10 ------Segment 11 ------3 12 ------13 ------

139

Table B-3: Results from the environmental DNA analysis for Black Carp from each sample collected at each sampling site in the Kansas River. Positive detections are denoted as (+) and negative detections as (-). Sites P and 1-3 were in Segment 1, sites 4-8 were in Segment 2, and sites 9-13 were in Segment 3. Samples 1 and 2 were collected near the South bank, samples 3 and 4 were collected from the thalweg, and samples 5 and

6 were collected from the North bank.

Sample Number Site 1 2 3 4 5 6 P + + + NA NA NA

Segment 1 ------1 2 ------3 ------4 ------5 ------Segment 6 ------2 7 ------8 ------9 ------10 ------Segment 11 ------3 12 ------13 ------

140

APPENDIX C

Supplemental tables and figures pertaining to Chapter 4

141

Table C-1: Results from the otolith microchemistry analysis on Silver Carp captured in

Segments 1 and 2 of the Kansas River. Unique ID is a unique identification code for each individual fish, Segment is the segment of capture, and Total length is the total length of the fish at capture. Silver Carp movement patterns were classified as either transient (i.e. had Sr:Ca signatures greater than 2,082 µmol/mol, indicative of the Missouri River at some point along the ablated transect) or resident if they lacked those signatures. Natal origins are the predicted natal origins associated with data from the nucleus of the otolith.

Natal origin signatures indicative of the Kansas were only from resident individuals.

Unknown origin individuals are transient fish that lack natal origin signatures indicative of the Missouri River. We were unable to predict natal origins other than the Missouri

River for transient individuals due to lack of otolith and water chemistry data throughout the Missouri River drainage.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180236-38 2 666 Transient Unknown 180199-6 2 644 Transient Unknown 180190-2 2 695 Resident Kansas 180192-1 2 710 Resident Kansas 180190-1 2 681 Transient Unknown 180237-1 2 605 Transient Missouri 180147-8 2 605 Resident Kansas 180192-2 2 708 Transient Unknown 180237-3 2 626 Resident Kansas 180239-16 2 608 Transient Unknown 180237-5 2 726 Resident Kansas 180237-4 2 668 Resident Kansas 142

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180237-4 2 668 Resident Kansas 180186-9 2 706 Transient Unknown 180190-3 2 682 Transient Unknown 180079-3 1 533 Resident Kansas 180235-57 2 671 Transient Unknown 180147-4 2 632 Resident Kansas 180239-18 2 671 Resident Kansas 180084-4 1 551 Resident Kansas 180147-6 2 646 Resident Kansas 180187-1 2 652 Transient Missouri 180056-3 1 588 Transient Unknown 180121-6 2 700 Resident Kansas 180147-9 2 590 Transient Missouri 180139-9 2 693 Transient Unknown 180067-2 2 690 Transient Missouri 180147-7 2 642 Transient Unknown 180139-7 2 685 Resident Kansas 180185-1 2 732 Transient Unknown 180199-5 2 648 Resident Kansas 180081-4 1 605 Transient Unknown 180239-15 2 671 Resident Kansas 180147-2 2 612 Transient Unknown 180121-5 2 731 Resident Kansas 180066-3 2 712 Resident Kansas 180233-2 2 645 Transient Unknown

143

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180139-10 2 786 Transient Missouri 180126-1 2 594 Resident Kansas 180082-2 1 531 Resident Kansas 180147-5 2 641 Resident Kansas 180067-3 1 756 Transient Unknown 180192-4 2 752 Transient Unknown 180237-2 2 694 Resident Kansas 180190-4 2 705 Resident Kansas 180292-12 1 558 Transient Unknown 180204-6 1 571 Transient Unknown 180199-4 2 696 Transient Unknown 180066-1 2 702 Transient Unknown 180239-17 2 637 Resident Kansas 180080-2 1 774 Transient Missouri 180080-1 1 660 Transient Missouri 180081-3 1 660 Transient Missouri 180056-6 1 512 Resident Kansas 180082-3 1 611 Resident Kansas 180081-2 1 575 Resident Kansas 180082-4 1 478 Resident Kansas 180241-3 2 645 Resident Kansas 180083-3 1 625 Transient Unknown 180141-10 2 656 Resident Kansas 180069-1 2 632 Resident Kansas 180204-8 1 562 Transient Unknown

144

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180093-3 1 777 Transient Unknown 180292-11 1 657 Resident Kansas 180090-1 1 755 Transient Unknown 180056-2 1 527 Transient Missouri 180082-5 1 629 Resident Kansas 180079-6 1 564 Transient Unknown 180291-2 1 613 Resident Kansas 180208-2 1 576 Transient Missouri 180291-3 1 661 Resident Kansas 180204-7 1 598 Transient Unknown 180292-8 1 605 Resident Kansas 180292-2 1 641 Transient Unknown 180203-17 1 578 Resident Kansas 180291-1 1 631 Transient Unknown 180292-6 1 520 Resident Kansas 180203-19 1 560 Resident Kansas 180208-6 1 562 Resident Kansas 180083-4 1 574 Resident Kansas 180292-5 1 662 Transient Missouri 180079-7 1 580 Transient Unknown 180208-4 1 567 Resident Kansas 180083-2 1 573 Resident Kansas 180147-10 2 561 Resident Kansas 180203-18 1 582 Resident Kansas 180139-1 2 711 Resident Kansas

145

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180121-3 2 694 Resident Kansas 180036-1 2 690 Resident Kansas 180151-4 1 674 Resident Kansas 180132-22 2 675 Resident Kansas 180139-2 2 677 Transient Unknown 180142-1 2 656 Transient Unknown 180142-3 2 647 Transient Unknown 180139-4 2 760 Resident Kansas 180121-2 2 755 Resident Kansas 180292-4 1 640 Transient Unknown 180132-23 2 670 Transient Unknown 180292-7 1 614 Resident Kansas 180209-1 1 605 Transient Unknown 180132-22 2 675 Resident Kansas 180151-5 1 675 Resident Kansas 180292-10 1 612 Resident Kansas 180130-1 2 577 Resident Kansas 180207-4 1 556 Resident Kansas 180121-1 2 700 Transient Unknown 180139-6 2 680 Resident Kansas 180122-3 2 699 Transient Unknown 180048-2 2 594 Resident Kansas 180139-3 2 713 Transient Unknown 180142-2 2 661 Transient Unknown 180291-4 1 654 Resident Kansas

146

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180151-6 1 645 Resident Kansas 180146-3 1 630 Transient Missouri 180079-5 1 626 Resident Kansas 180080-4 1 562 Resident Kansas 180153-5 1 612 Transient Unknown 180151-1 1 645 Transient Unknown 180153-2 1 611 Transient Unknown 180151-7 1 521 Resident Kansas 180132-21 2 646 Resident Kansas 180151-10 1 666 Transient Unknown 180151-9 1 653 Transient Missouri 180151-2 1 675 Transient Unknown 180208-12 1 732 Resident Kansas 180151-8 1 709 Transient Unknown 180153-1 1 609 Resident Kansas 180080-3 1 500 Resident Kansas 180153-4 1 579 Resident Kansas 180146-5 1 628 Transient Unknown 180147-1 2 605 Resident Kansas 180151-3 1 701 Transient Missouri 180192-3 2 682 Transient Missouri 180098-1 2 655 Transient Unknown 180080-5 1 618 Transient Missouri 180121-4 2 732 Transient Unknown 180147-11 2 604 Resident Kansas

147

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 180081-5 1 512 Transient Unknown 180130-3 2 695 Transient Unknown 180139-5 2 741 Transient Unknown 180130-2 2 660 Transient Unknown 180204-5 1 741 Resident Kansas 190358-16 2 590 Resident Kansas 190504-4 1 291 Resident Kansas 190504-5 1 278 Transient Unknown 190504-3 1 295 Transient Unknown 190503-6 1 304 Resident Kansas 190366-9 2 646 Transient Unknown 190117-6 2 546 Resident Kansas 190323-2 2 660 Resident Kansas 190366-7 2 686 Transient Unknown 190323-1 2 842 Resident Kansas 190323-4 2 643 Transient Unknown 190117-8 2 708 Resident Kansas 190211-10 2 602 Resident Kansas 190303-5 2 210 Resident Kansas 190116-12 2 646 Resident Kansas 190538-20 1 286 Resident Kansas 190303-2 2 206 Resident Kansas 190512-7 1 285 Resident Kansas 190303-4 2 210 Resident Kansas 190214-21 1 576 Resident Kansas

148

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 190397-20 1 129 Resident Kansas 190503-9 1 281 Transient Unknown 190366-2 2 635 Transient Unknown 190538-14 1 311 Resident Kansas 190366-6 2 700 Transient Unknown 190214-24 1 721 Resident Kansas 190220-2 1 530 Resident Kansas 190397-11 1 575 Resident Kansas 190521-3 2 678 Resident Kansas 190397-7 1 561 Transient Unknown 190397-18 1 117 Transient Missouri 190397-3 1 588 Transient Unknown 190397-10 1 139 Transient Missouri 190397-9 1 277 Resident Kansas 190214-9 1 800 Resident Kansas 190524-3 2 641 Transient Unknown 190153-36 1 110 Resident Kansas 190153-37 1 130 Transient Missouri 190153-40 1 124 Resident Kansas 190153-41 1 130 Resident Kansas 190356-2 2 630 Transient Unknown 190167-4 2 651 Transient Unknown 190502-14 1 279 Transient Missouri 190502-11 1 284 Transient Unknown 190218-1 1 575 Resident Kansas

149

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 190142-8 1 524 Resident Kansas 190311-13 1 582 Resident Kansas 190142-16 1 271 Resident Kansas 190307-2 2 738 Transient Unknown 190397-1 2 559 Resident Kansas 190355-1 2 385 Transient Missouri 190222-4 2 645 Transient Unknown 190339-4 1 590 Resident Kansas 190222-3 2 565 Resident Kansas 190222-5 2 729 Resident Kansas 190135-3 1 614 Transient Unknown 190222-2 2 660 Resident Kansas 190222-18 2 224 Transient Unknown 190337-10 1 602 Resident Kansas 190355-6 2 620 Transient Unknown 190364-6 2 564 Transient Missouri 190305-2 2 695 Resident Kansas 190363-2 2 622 Resident Kansas 190308-1 2 626 Transient Unknown 190610-15 2 580 Transient Unknown 190602-4 2 543 Transient Unknown 190531-4 2 701 Transient Unknown 190153-39 1 120 Transient Missouri 190159-30 1 116 Resident Kansas 190530-8 2 625 Resident Kansas

150

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 190540-9 1 300 Resident Kansas 190160-6 2 568 Resident Kansas 190503-7 1 303 Transient Unknown 190511-6 1 296 Resident Kansas 190602-9 2 550 Resident Kansas 190602-15 2 690 Resident Kansas 190164-2 2 585 Resident Kansas 190117-3 2 520 Resident Kansas 190346-4 1 550 Resident Kansas 190535-9 1 293 Resident Kansas 190159-16 1 128 Resident Kansas 190252-15 2 684 Transient Unknown 190252-10 2 732 Transient Unknown 190271-25 2 605 Resident Kansas 190078-3 2 644 Resident Kansas 190078-19 2 675 Transient Unknown 190241-7 2 685 Transient Missouri 190252-18 2 540 Resident Kansas 190168-2 2 653 Transient Unknown 190159-18 1 118 Resident Kansas 190540-8 1 346 Transient Unknown 190164-4 2 643 Resident Kansas 190153-35 1 121 Resident Kansas 190158-29 1 261 Resident Kansas 190525-2 2 620 Resident Kansas

151

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 190612-2 2 635 Resident Kansas 190531-5 2 705 Transient Unknown 190161-5 2 665 Resident Kansas 190525-3 2 641 Resident Kansas 190525-4 2 647 Resident Kansas 190252-19 2 625 Resident Kansas 190251-9 2 660 Resident Kansas 190261-1 2 652 Transient Unknown 190073-15 2 680 Resident Kansas 190262-2 2 565 Resident Kansas 190232-1 2 621 Transient Unknown 190271-23 2 654 Resident Kansas 190233-5 2 631 Transient Unknown 190084-4 2 692 Resident Kansas 190271-7 2 680 Transient Unknown 190271-16 2 715 Resident Kansas 190252-11 2 650 Transient Unknown 190078-21 2 721 Resident Kansas 190241-8 2 650 Transient Unknown 190266-4 2 665 Transient Unknown 190241-2 2 676 Resident Kansas 190078-23 2 708 Transient Missouri 190073-18 2 625 Resident Kansas 190268-33 2 625 Transient Unknown 190259-2 2 638 Transient Unknown

152

Table C-1: Continued.

Unique ID Segment Total length (MM) Movement pattern Natal origins 190023-7 2 755 Transient Unknown 190030-4 2 540 Resident Kansas 190046-2 1 667 Transient Unknown 190035-25 2 670 Resident Kansas 190035-31 2 649 Resident Kansas 190035-32 2 618 Resident Kansas 190056-19 1 645 Resident Kansas 190046-1 1 676 Resident Kansas 190035-29 2 666 Transient Unknown 190044-16 1 545 Resident Kansas 190078-25 2 633 Resident Kansas 190080-8 2 670 Transient Unknown 190268-26 2 612 Transient Unknown 190511-5 1 273 Resident Kansas 190153-34 1 122 Resident Kansas

153

Figure C-1: Recursive partitioning tree using Ba:Ca signatures to classify environmental history of Silver Carp between the Kansas River and the Missouri River. The tree was pruned at size = 2 with a complexity parameter (cp) of 0.071.

154

Figure C-2: Number of laser ablation points above 2,082 µmol/mol Sr:Ca theshold for each individual transient fish by total length. Data points greater than 2,082 µmol/mol

Sr:Ca were indicative of Missouri River trace element signatures in lapilli otoliths of

Silver Carp.