I quantified habitat selection for the endangered Zuni Bluehead Sucker Catostomus discobolus yarrowi and the Navajo Nation Genetic Subunit (NNGS) Bluehead Sucker Catostomus discobolus - a recent taxon described from genetic information. Both taxa are found in northern Arizona and New Mexico border regions. I examined fish [≥50 millimeters (mm) total length (TL)] selection of microhabitat conditions (i.e., water velocity, substrate size, overhead cover, water depth, instream cover, and mesohabitat conditions [i.e., pool, run riffle], during summer base flow conditions for NNGS Bluehead Suckers, and during both summer base flow and high spring flow conditions for Zuni Bluehead Suckers in six streams). Electrofishing, seining, and snorkeling were used to evaluate fish occupancy. From this information, I developed stream specific habitat suitability criteria (HSC) and then generalized HSC for each taxon, and tested transferability of the generalized HSC to individual streams. Zuni Bluehead Suckers and NNGS Bluehead Suckers occupied similar habitats: low velocity pools; sand, silt, and pebble substrate; high percent of instream cover; and water temperatures ranging from 2-21°C. However, Zuni Bluehead Suckers selected for low (0-25%) overhead cover where as NNGS Bluehead Sucker selected for high (0-75%) overhead cover. This was likely due to the source of instream cover–aquatic macrophytes that required sunlight in the Zuni Bluehead Sucker streams, and large woody debris falling from overhead branches in the NNGS Bluehead Sucker streams. Suggestions for managers includes maintaining existing cover or artificially construct additional instream cover; promote overhead cover (e.g., maintaining large trees along streams) and pool mesohabitats. In addition to this work I also tested the new method of environmental DNA (eDNA) to further help conservation efforts for these taxa. Environmental DNA has typically been used to detect invasive species in aquatic environments through water samples. I compared the efficacy of eDNA methodology to American Fisheries Society standard snorkeling surveys to detect presence of a rare fish species. My study site included three streams on the Navajo Nation in northern Arizona and northern New Mexico containing Navajo Nation Genetic Subunit Bluehead Sucker Catostomus discobolus and the Zuni Bluehead Sucker Catostomus discobolus yarrowi. To determine sample sites, I first divided entire wetted area of streams into 100-m consecutive reaches. I systematically selected 10 of those reaches for snorkel and eDNA surveys. Water samples were taken in 10-m sections within each 100-m reach, and fish presence via snorkeling was noted in each 10-m section as well. Water samples were collected at the downstream starting point of each reach, and continued upstream in each section 5 to 8 m ahead of the snorkeler. A qPCR was run on each individual water sample in quadruplicate to test for sucker presence or absence. I was able to positively detect both species with eDNA sampling techniques in two out of three streams. Snorkeling resulted in positive detections of both species in all three streams. In streams where fish were detected with eDNA sampling, snorkeling detected fishes at 11-29 sites per stream, where as eDNA detected fish at 3-12 sites per streams. My results suggested that AFS standard snorkeling was more effective at detecting target fish species than eDNA. To improve eDNA sampling, the amount of water collected and tested should be increased. Additionally, filtering water on site may improve eDNA techniques for detecting fish. Future research should focus on standardizing eDNA sampling to provide a widely operational sampling tool similar to electrofishing, netting, and hydroacoustics.

With a new focus on flow regulation by the Tennessee Valley Authority (TVA) in reservoir tailwaters, it is now possible to recover many mussel species that once occurred in these reaches. Before flows can be modified to create habitat for freshwater mussels, suitable microhabitat conditions must be defined. In this study, I used multiple approaches to define suitable microhabitats for species in the free-flowing upper Clinch River, Virginia and Tennessee, where reproducing mussel populations persist. During summer low flows in 2003 and 2004, I measured flow and substrate conditions in over 1000 microhabitat patches (0.25 m² quadrat samples) across five river reaches. Flow characteristics and embeddedness were significantly different between microhabitats occupied and unoccupied by the most abundant species (MRPP, p < 0.05). Comparison of simple and multiple logistic regression models with Akaike's Information Criteria (AIC) demonstrated that increasing Fleisswasserstammtisch (FST) hemisphere number (a measure of shear stress), decreasing degree of embeddedness, and increasing mean column velocity best explained species occurrences in a microhabitat patch. Subtle differentiation in habitat use among species was observed; however, most species appeared to be microhabitat generalists. Species were grouped into three habitat guilds using corresponding canonical analysis and cluster analysis: fast-flow specialists (FFS), fast-flow generalists (FFG), and slow-flow tolerant (SFT). I used the same data set to develop and test transferability of Habitat Suitability Criteria (HSC) for three habitat guilds and seven species of adult freshwater mussels. Nonparametric tolerance limits were used to define the range of suitable and optimal habitat during summer low flows. Optimal habitat was defined as those ranges of FST hemisphere number, mean column velocity, and embeddedness occupied by the central 50% of independent observations for a species or guild, whereas suitable habitat was defined by those ranges occupied by the central 90% of observations. The transferability of criteria to other reaches of the Clinch River was assessed using one-sided Chi-square tests. Criteria developed for the fast-flow specialist (FFS) and fast-flow generalist (FFG) guilds, as well as most criteria for species in those guilds, transferred to destination reaches. In contrast, criteria developed for the slow-flow tolerant (SFT) guild and individual constituent species consistently failed to transfer. Criteria for FFS and FFG guilds and their constituent species should be incorporated into flow simulation models such as PHABSIM to gauge the effect of minimum flows on mussel habitat quality and quantity. These criteria could also be used to determine suitable sites for mussel translocations. However, my criteria require further testing in other rivers before they can be transferred beyond the Clinch River. Behavior and physiological responses to laboratory manipulations of flow velocity and substrate particle size were used to elucidate microhabitat preferences of Actinonaias pectorosa, Potamilus alatus, and Ptychobranchus subtentum. These species appeared less stressed in the fastest flow treatment, demonstrating significantly higher oxygen consumption and oxygen-to-nitrogen (O:N) ratios than in slower flow treatments. Only P. alatus demonstrated a preference for substrate particle size, and consistently selected finer particle sizes. Actinonaias pectorosa and P. subtentum demonstrated preference for fast-flow microhabitats by readily burrowing in those conditions, while abandoning slow-flow conditions. The lack of preference for substrate particle size demonstrated by A. pectorosa and P. subtentum supports conclusions of previous studies that substrate particle size is of little or secondary importance for explaining mussel microhabitat use. These results, along with previous studies in the Clinch River, demonstrate that the stable habitats of riffles and runs; characterized by fast flows during summer low flows, low percent bedrock, and low embeddedness, are the most suitable habitats for mussel assemblages. To create and maintain suitable habitat conditions in tailwaters, releases should maintain flow over riffles at a minimum depth of no less than 30 cm in riffles that provide higher shear stress conditions (FST number > 7) and velocities (> 0.70 m/s). Periodic releases that are sufficient to transport silt and sand, but not high enough to transport larger substrate should be adequate to maintain substrates with a low degree of embeddedness. Doing so would create suitable habitat for all mussels, from the most to least specialized. Additionally, HSC developed for FFS and FFG guilds can be used to determine suitable destination sites for translocations of species belonging to these guilds. / Master of Science

This study constructed habitat suitability criteria for fishes of the South Fork of the Shenandoah River, in Virginia. The criteria will be used in an IFIM study to produce estimates of the discharge required by fishes in the South Fork. Chi-square tests were used to evaluate whether criteria described habitat use to a statistically significant degree. Secondly, chi-square tests were used to test transferability. The criteria described the habitat use of seven taxa commonly found in the South Fork to a statistically significant degree. Habitat criteria for two taxa did not describe their habitat use to a statistically significant degree. One set of criteria from the North Fork of the Shenandoah transferred to the fish observed in the South Fork. Secondly, this paper examined observer effects of underwater observation. It was of interest to explore how observer effects influenced habitat suitability criteria.

The diversity of fish species found in warmwater stream systems provides a perplexing challenge when selecting species for Instream Flow Incremental Methodology (IFIM) studies. An often-suggested approach has been to use habitat guilds to incorporate the diversity found in these systems. My goal is to determine the feasibility of developing habitat suitability criteria (HSC) for the entire fish assemblage in the North and South Fork Shenandoah River, Virginia, using habitat guilds. I examined the strengths and weaknesses of direct underwater observation via snorkeling and throwable anode electrofishing to sample fish habitat use (e.g., depth, velocity, distance to cover, dominant and subdominant substrate, cover, and embeddedness) indicates that using the data collected from both techniques may produce better criteria than using just one of the two sampling techniques. To develop habitat suitability criteria using habitat guilds I placed each species a priori into a guild based on a hypothesized guild structure. Transitional life stages with significantly different habitat use were placed separately into the guild structure. The four guilds (riffle, fast generalist, pool-run, and pool-cover) were found to be significantly different from each other using the data collected for the species assigned to the guilds. Criteria were then developed for representative species from each guild and the entire guilds. Criteria developed for depth, velocity, Froude number, cover, distance to cover, substrate, and embeddedness were used to estimate a habitat response function (i.e., the relations between usable habitat and stream flow) for a representative species from each guild, the guild itself, and for a second species from each guild for comparisons. Both the representative species and guild criteria showed similar habitat response functions for the riffle guild, fast generalist guild, and pool-run guild. However, neither set of criteria performed well for the pool-cover guild. For guilds, other than pool-cover, either the guild or the representative species approach may be a viable option to developing habitat suitability criteria. The transferability tests were performed to determine if criteria developed in the North Fork Shenandoah River, Virginia would transfer to the South Fork Shenandoah River, Virginia. Only criteria for the margined madtom (Noturus insignis) and the juvenile smallmouth bass (Micropterus dolomieu) transferred for both suitable and optimal habitat. Criteria for mottled sculpin (Cottus bairdi), Cyprinella sp. (spotfin and satinfin shiners), river chub (Nocomis micropogon), adult and juvenile redbreast sunfish (Lepomis auritus), and adult smallmouth bass did not transfer. Only the pool-cover guild criteria transferred for both suitable and optimal habitat, while riffle guild, fast generalist guild, and pool-run guild criteria did not transfer. I recommend the use of site-specific criteria for the South Fork Shenandoah or different variable combinations. / Master of Science