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FRESHWATER MUSSELS OF THE UPPER SAN ANTONIO RIVER WATERSHED AND LOWER CIBOLO CREEK IN BEXAR, GUADALUPE, WILSON, AND KARNES COUNTIES: ABUNDANCE AND DENSITIES.

Report By

Larry Larralde San Antonio River Authority

December 2016

Table of Contents

ABSTRACT ...... 1 INTRODUCTION ...... 2 DECLINING FACTORS ...... 2 SARA BASELINE GOALS ...... 3 WESTSIDE CREEKS ...... 3 LOWER CIBOLO CREEK ...... 6 METHODS AND MATERIALS ...... 8 DEEP POOL HABITAT: QUANTITATIVE ...... 10 DEEP POOL HABITAT: QUALITATIVE ...... 11 OTHER MESOHABITATS QUANTITATIVE ...... 11 OTHER MESOHABITATS QUALITATIVE ...... 13 OTHER MEASUREMENTS ...... 13 WESTSIDE CREEKS ...... 14 QUALITATIVE RESULTS ...... 14 VELOCITIES AND DEPTHS, WESTSIDE CREEKS ...... 14 LOWER CIBOLO CREEK RESULTS ...... 15 QUANTITATIVE ...... 16 BIOLOGICAL STRUCTURE ...... 18 QUALITATIVE ...... 19 VELOCITIES AND DEPTHS ...... 22 POPULATION STRUCTURE ...... 23 ASSOCIATED MUSSEL MESOHABITATS ...... 27 GEOSPATIAL CONTRAST ...... 28 LITERATURE CITED ...... 29 APPENDIX A: WATER QUALITY AT SELECTED SITES...... 32 WESTSIDE CREEKS, WATER QUALITY DATA, TCEQ SEGMENT 1911 ...... 32 APACHE CREEK AT BRAZOS ...... 33 MARTINEZ CREEK AT RUIZ ...... 35 ALAZAN CREEK AT TAMPICO ...... 37 SAN PEDRO CREEK AT MITCHELL ...... 39 LOWER CIBOLO CREEK, WATER QUALITY DATA, TCEQ SEGMENT 1902 ...... 41 CIBOLO CREEK AT CR 389 ...... 41

APPENDIX B: LOCATION, NUMBER AND TYPES OF MUSSELS FOUND...... 44 APPENDIX C: FOCUS ON GOLDEN ORB MUSSELS ...... 46 SPECIAL THANKS: ...... 52

LIST OF FIGURES:

Figure 1. Martinez Creek ...... 3 Figure 2. Apache Creek ...... 4 Figure 3. Alazan Creek ...... 4 Figure 4. Westside Creeks and Lower Cibolo Creek Watershed ...... 5 Figure 5. Lower Cibolo Creek ...... 6 Figure 6. Lower Cibolo Creek Watershed Collection Sites, TCEQ Segment 1902...... 7 Figure 7. Lower Cibolo Creek Watershed Sample Reaches ...... 9 Figure 8. 0.25m2 Quadrat with sock...... 10 Figure 9. Panning with Sieves ...... 11 Figure 10. Matrix Grid ...... 12 Figure 11. USGS Hydrograph ...... 15 Figure 12. USGS Hydrograph ...... 15 Figure 13. Mussels ...... 19 Figure 14. Pistolgrip Size Classes ...... 25 Figure 15. Yellow Sandshell Size Classes ...... 25 Figure 16. Golden Orb Size Classes ...... 26 Figure 17. Louisiana Fatmucket Size Classes ...... 26 Figure 18. Paper Pondshell Size Classes ...... 27 Figure 19. Mussel Mesohabitats Relative to Mussel Abundance ...... 28 Figure 20. Relative Spatial Reaches on Mussel Abundance ...... 29 Figure 21C Locations of Golden Orb Mussels ...... 46

LIST OF TABLES:

Table 1. Sample Reaches, Lower Cibolo Creek ...... 8 Table 2. Deep Pool Sample Sites ...... 10 Table 3. Mussel Species, Search Area m2, and CPUE's, Westside Creeks ...... 14 Table 4. Velocities and Depths (Mean ± SE), Westside Creeks ...... 15 Table 5. Quantitative Mussel Data, Lower Cibolo Creek by Sample Reaches ...... 17 Table 6. Indices of Evenness and Diversity by Reaches, Upstream to Downstream ...... 19 Table 7. Qualitative Mussel Data, Lower Cibolo Creek ...... 21 Table 8. Deep Pooled Mussel Sample Sites and Results ...... 22 Table 9. Velocities and Depths (mean ± SE), Lower Cibolo Creek ...... 23 Table 10. Impairment and concerns from 2014 Integrated Report...... 32

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ABSTRACT

Three mussel species under review for federal listing with the United States Endangered Species Act (ESA) and legally threatened in the State of Texas have historically been found in the San Antonio River (SAR) basin. One species, golden orb Quadrula aurea has been recently found in numerous locations in the San Antonio River and its tributaries (Lower Cibolo Creek). Previous efforts to survey the lower Cibolo Creek watershed began in the early 1990’s and no records have been located regarding the westside creeks (San Pedro, Alazan, Apache and Martinez Creeks just west of downtown San Antonio). Therefore, the San Antonio River Authority (SARA) decided to conduct a basin wide monitoring efforts for mussels. Sample data included baseline demographics such as density, richness, diversity, and age-class structure for the native mussel community. Data will be collected and distributed to regulatory agencies to assist in the decision making process for listing or delisting candidate species of concern. Sample efforts for 2014-2015 included the westside creeks of the Upper San Antonio Watershed from the Texas Commission on Environmental Quality (TCEQ) segment 1911, and the Lower Cibolo Creek, TCEQ segment 1902. Quantitative sample methods (Systematic Sampling) were implemented with a 0.25m2 quadrat and catchment along with qualitative timed catch-per-unit-effort (CPUE) expressed as number of mussels found per hour (no. /h) where visual and tactile searches were conducted at 46 collection sites across five lower Cibolo Creek sample reaches. Qualitative catch-per-unit-efforts under visual timed searches was the preferred and only method utilized at 40 sample sites for the westside creeks. Qualitative results from all four westside creeks yielded one native mussel species, paper pondshell Utterbackia imbecillis, from Martinez Creek. The CPUE for Martinez Creek was 0.83/h., and overall, five live specimens were collected with a CPUE of 0.26/h mussels for all four creeks combined. Quantitative results from lower Cibolo Creek yielded a total of 81 observed individuals represented by five native Unionid species, yellow sandshell Lampsilis teres (n=38), golden orb Quadrula aurea, (n=32), paper pondshell Utterbackia imbecillis (n=3), pistolgrip Quadrula verrucosa (n=7), and louisiana fatmucket Lampsilis hydiana (n=1). The quadrat range per sample site was 13-17 with a total of 720 quadrats for all five sample reaches. Only one fourth (26%) of our overall quantitative sample had measurable data indicative of low mussel densities and patchy mussel distributions from the sample area that ranged from 0.0m2 – 0.96m2. The quantitative mean mussel density for all sites was 0.152m2 ± 0.01 and seemed to be the standard. In this study, Qualitative timed searches were more successful than quantitative and exhibited mussel presence 52% of the time. Qualitative results produced an overall CPUE range of 0.0 – 2.33 /h and a mean CPUE for the community from all sites of 0.40/h ± 0.08. Our species of conservation concern, golden orb, was collected at 35% of all our sample sites in low densities and represented by thirty - two individuals. The mean CPUE for golden orb for all sites was 0.13/h ± 0.05. Statistical analyses revealed species richness in quadrat samples relative to species richness in timed searches were found to be significantly correlated (Spearman’s rho, r=0.36, P = 0.01). Also, a Paired 2 Sample t-test was ran to test the Null hypothesis comparing the effectiveness of quantitative (quadrat) and qualitative (timed searches) mussel surveys and results were significant at the <.05 level (P = 0.008 ). Shell lengths, anterior to posterior were measured and collected to provide insight on community size class structure, however, class estimates could be biased due to a small sample size. Louisiana fatmucket and pistolgrip lacked representative juveniles, and conversely all other species except paper pondshell were represented by medium to large sized adults. Golden orb was represented by only a few juveniles and more representative in the 31- 40mm class. The largest class was represented by pistolgrip, yellow sandshell, and golden orb in moderation, however, there are also some recent recruitment issues from several species. Habitat

utilization was dominated by golden orb and yellow sandshell located in glide/run combinations. There also seemed to be a disjointed mussel community where no individuals were located in the farthest upstream reach and just below the middle reach where mussel abundance rose, slowly declined, and then finally rose. The lower Cibolo Creek mussel community has high species richness however densities are relatively low and evenness among species is lacking. The Golden orb is a state threatened mussel found in relatively low abundance in the lower Cibolo Creek, a tributary to the San Antonio River.

INTRODUCTION Freshwater mussel populations have recently become the most studied groups of organisms across North America due to the alarming rate of their declining numbers. Many species have been locally extirpated; others, which are scarce, have been listed at state or federal levels of conservation; and some are already extinct through habitat destruction (Bogan 1993; Williams et al. 1993). Mussel species are one the most rapidly declining faunal groups in North America (Turgeon et al.,1998) mostly influenced by human activities with many species already extinct and many more in dire need of conservation. The creation of dams alters the hydraulic flow regime in rivers which reduces water flow, accumulates silt, and increases water fluctuations reducing mussel fauna (Vaughn and Taylor, 1999). Flow regimes associated with dam operations, along with nonpoint source pollution, and invasives are three leading causes of the imperilment of aquatic organisms (Richter et al. 2003). Since these organisms play an important role in water quality and can serve as useful monitors of water pollution (Green et al., 1989; Metcalfe and Charlton 1990), they have been the subject of many recent studies throughout watersheds across North America. Through physiological inherited traits their mechanical means of filter feeding subjects them to a wide range of microorganisms and toxic compounds where sensitivity to toxins, compounded by limited mobility and a long life span make unionids extremely vulnerable (Fuller, 1974). In Texas, there are approximately 52 species that occupy river ways, marshes, reservoirs, and bayous collectively. The Texas Parks and Wildlife Department (TPWD) has listed 15 species as threatened with five labelled as Central Texas species, three of which are identified as located in the San Antonio River (SAR) basin. Currently, these species are also listed as Federal candidates by the United States Fish and Wildlife Service (USFWS) where a proposed ruling (or withdrawal) will be evaluated and concluded after Fiscal Year 2016.

DECLINING FACTORS Overall, mussel loss can be attributed to many factors within riverine systems: (1) habitat destruction and alteration, (2) fragmentation of waterways, e.g., dams, weirs, and impoundments which alter flow regimes and interrupt life reproductive cycles, (3) high sediment loads that choke and bury larvae and adults, (4) chemical pollutants, (5) scouring of bed channels from peak pulse flow events, (6) dewatering and chronic water level fluctuations, and (7) competition from invasive species.

SARA BASELINE GOALS SARA goals and objectives will be to determine and assess mussel population(s) throughout the basin and in particular for this report the lower Cibolo Creek Watershed and the urban westside creeks of the upper San Antonio River Watershed (Martinez Creek, San Pedro Creek, Apache Creek, and Alazan Creek) for native Unionids or signs of remnant shell fragments. Sampling for these taxonomic groups has never been conducted on the westside creeks while several surveys have been conducted along the lower Cibolo Creek (Howells 1995, 1997a; Karatayev and Burlakova 2008, SARA 2011). Since sampling for mussels in the basin has been sporadic and due to the recent interest in mussel decline across North America, the SARA proactively decided to begin sampling watersheds and sub-watersheds across the SAR basin. The SARA intends to make quantitative inferences about the mussel community in general where SARA biologists will assess mussel community conditions based on an array of population parameters. The data will be provided to other regulatory agencies at the state and federal level as they make recommendations for species of conservation concern within the SAR Basin.

This study does not focus on a single target species but instead looks at the entire mussel community assemblage by defining population parameters such as size, richness, and density and include other population demographics for all species observed and collected. A habitat utilization analysis will provide insight between mesohabitat types and mussel location, and geospatial differences in mussel abundance between upstream and downstream reaches. SARA biologists will also measure shell lengths to determine population size class structure, larval recruitment, and voids in the mussel community as a whole.

WESTSIDE CREEKS – Upper San Antonio Watershed, Texas Commission Environmental Quality, TCEQ Segment 1911 B, C, D, and I The westside creeks of the upper San Antonio River Watershed (Segment 1911) within Bexar County can be characterized as small urbanized creeks that converge flows into one another that eventually drains into the San Antonio River. The total length of each creek ranges from 1 – 4.75 km. These small sub-watersheds are considered First Order streams (Strahler, 1952) where channel streambeds are very narrow (≤ 3 meters, Figure 1. Martinez Creek field observations) and depths shallow. Riparian buffers are moderately wide for the most part (Figures 1, 2, and 3), densely vegetated, and maintained while the canopy cover is usually open and cover is lacking. The substrate consists of heterogeneous mixtures of large and small gravel, soft or hard clay and their loams, bedrock and small boulders, silt and accumulated floating and

non-floating assorted trash. All four tributaries (Figure 4) are considered urban and are located adjacent to commercial business establishments or residential housing along the longitudinal length of each stream, typical of urban streams. During peak rainfall events these creeks can rise quickly from sheet runoff due to nearby impervious cover and, conversely, can become very dry during long hot drought-like conditions where filamentous algae becomes denser and the creeks start to pool up. To SARA’s knowledge, these creeks have never been surveyed for freshwater mussels. This project effort will link up to both the San Pedro Creek Improvement Project, where hike and bike linear trails will be created, and to the upcoming westside creeks Project, where the streams and their riparian zones will be restored utilizing a Natural Stream Design.

Figure 2. Apache Creek Figure 3. Alazan Creek

Figure 4. Westside Creeks and Lower Cibolo Creek Watershed

LOWER CIBOLO CREEK, TCEQ SEGMENT 1902 The lower Cibolo Creek (SARA, 2008) begins at its confluence with the San Antonio River in Karnes County to a point approximately 100 meters downstream of the IH – 10 interstate in Bexar/Guadalupe County (Figure 6). The segment is 71 miles long and flows southeast over the Texas Blackland Prairie then over the east Central Texas Plains near the Bexar/Guadalupe County Line. Then it flows southeast until it reaches the confluence to the San Antonio River near Panna Maria, Texas, in Karnes County. The topography in these reaches is considered steep terrain that tapers to lower rolling hills heading further south. The banks are somewhat steep and vegetated typical to other streams within the vicinity. Riparian vegetation associated with dense hardwood can be found throughout the segment in various densities along the toe of the streambank, and transitioning into upland zone from the riparian zone into the floodplain. Canopy cover is generally present throughout the corridor and provides for good cover and habitat for fish and avian species. High and medium storied canopies vary along Cibolo Creek where cover can be completely closed or completely open depending on location within segment 1902. Flow within segment 1902 begins southwest of the city of Schertz as a combination of either spring flow, treated effluent, or runoff from storm water rain events. There are five wastewater treatment plants located in this segment’s watershed that discharge effluent which augments natural flow regimes in this segment. All habitat types are generally present but usually the lower Cibolo Creek is dominated by pools and glides with slower water velocities (Figure 5). Erosion and deposition along the creek are significant especially in reaches where the creek meanders sharply, and cut banks are extremely high. The substrate consists of heterogeneous mixtures of large and fine sand, silt, detritus, clay fines, and large sandstone depending on the location. Log jams are a common occurrence in the creek providing various macro and micro habitat types for aquatic invertebrates and nekton species.

Figure 5. Lower Cibolo Creek

Figure 6. Lower Cibolo Creek Watershed Collection Sites, TCEQ Segment 1902.

METHODS AND MATERIALS After receiving input from state agencies through personal communication or via- teleconference regarding sample design for this project, SARA implemented a Quantitative Systematic Sampling Design using 0.25m2 quadrats (Figure 8) from Strayer and Smith (2003) along with a timed qualitative catch-per-unit-effort (CPUE, no. /h) at each site allowing for maximum search sample efforts. In previous discussions with TPWD staff (2014) regarding mussel habitat types, both agencies collaborated about several topics which included sampling various mesohabitat types throughout every reach and the mechanics of sampling utilizing the systematic sample design as a primary sample tool. Similarly, SARA staff had several discussions with field experts at the Texas A & M Institute of Renewable Natural Resources, both in person and via teleconference in 2014 which resulted in a recommendation that the quantitative (systematic) sample design be accompanied with a qualitative effort. Sample reach and site selection (Table 1) were somewhat dependent upon logistics due to the area within the watershed (Figure 7) where accessibility was available, and field safety a priority, and where SARA biologists completed pre-reconnaissance efforts at potential reaches before selecting reaches and sites. Selected reaches followed specific criteria where sample reaches were no longer than 5 miles in length from entry to exit point (sample reach distance), geospatially and evenly distributed, and where habitat consisting of heterogeneous mixtures of small and large sand, clay, silt, mud, and gravel occurred. Sites within were selected based on best professional judgment. Once a number of sites were selected, that number was doubled (2X), and the reach divided into evenly spaced sites. We felt justifying the number of sites times two to ensure that at least 50% of the sites within a reach would be sampled. Sites were then numbered accordingly 1-2-3 X etc. where site selection was done randomly from a computer based website. If a reach had 10 sites spaced evenly then 5 were chosen and placed into a GPS unit to refer to in the field. Upon arrival at that point, if the site was too deep for wading or flows were unsafe to sample then the field crew kayaked farther downstream until a more workable area was located.

Table 1. Sample Reaches, Lower Cibolo Creek Sample Reach Number

Sites FM 2538 – Zuehl Crossing (CR 440) 11 Scull Crossing – FM 775 9 FM 775 – CR 337 6 FM 537 – CR225 7 State Hwy. 123 – FM 2724 13

Figure 7. Lower Cibolo Creek Watershed Sample Reaches Sample sites (Table 1) were selected randomly along the lower Cibolo Creek (46 sites) and the West Side Creeks (40 sites), where the focus of pre – reconnaissance efforts were to locate mussel habitat, and signs of extant mussel communities thus increase the likelihood of locating live mussels. For this study, mesohabitat types were labelled as glide/run combinations, riffles, backwater, and pools. Once collected, raw data from every site was recorded, graphed, and analyzed accordingly to visualize mussel abundance across sample reaches. Ambient weather conditions during FY 2014 spring sample (Figures 11 and 12) events were wetter than normal producing rains throughout the region which impacted discharge rates thus delaying our sample efforts until flows were safer to sample in. Deeper water and swifter velocities often made it challenging, where sampling was often restricted to the banks or as far to the centroid as possible. The United States Geological Survey website for flows was analyzed for the periods between 10/22/2014 through 8/22/2015. Sample methods for this study utilized two techniques separately but in conjunction: (1) implement qualitative visual/tactile surveys, panning substrate just below the surface at least

15cm or until firm, and utilize mussel sieves (Figures 8 and 9) if ambient conditions are inhibited by water clarity or depth. The qualitative (CPUE no. /h) method provides for estimates of species richness and can cover more area while, (2) Quantitative quadrat sampling provides for more precise estimates of density and size class structure (Vaughn et al. 1997; Strayer and Smith, 2003). Since these sample methods are different and produce vital information regarding mussel abundance in various ways, qualitative and quantitative raw data were kept and analyzed separately. Statistical analyses compared and examined sampling bias between quantitative (sediment excavation) and qualitative (timed searches) methods where a Paired 2 Sample t - test (Burlakova et al. 2011) at α = 0.05 was calculated based on the Null hypothesis stating there are no differences between quantitative and qualitative mussel sample densities. A correlation relative to both sample methods was also examined, and biological structure regarding diversity and evenness across reaches was analyzed. Methods for determining mussel assemblage (size class structure) at each reach was analyzed in two ways: visual / tactile and quadrat sampling. These methods were used because visual surveys cover more area, and quadrat sampling is unbiased relative to mussel size. Since a species of conservation concern (golden orb) has been already detected, efforts were focused at increasing the probability of locating not only larger specimens but also recent recruitment. DEEP POOL HABITAT: QUANTITATIVE

Table 2. Deep Pool Sample Sites Lower Cibolo Creek Pool # Location Coordinates 1 FM 2538 29.45148 -98.12305 2 Scull Crossing 29.39048 -98.12568 3 Scull Crossing 29.38952 -98.12447 4 SH 123 29.01452 -97.92937 5 CR 389 29.01484 -97.91898 Figure 8. 0.25m2 Quadrat with sock

Deeper pooled waters that are often turbid and sometimes difficult to access have shown to be prime habitat that can often support an abundant and diverse Unionid assemblage. Therefore, the SARA contracted with Bio-West Inc. to provide a third lung hookah team to conduct deep pool sampling efforts at five sites (Table 2). SARA field staff and Bio-West Inc. set up a minimum of 5 perpendicular transect tag lines per site which were stretched and secured across the channel. Four random generated points were chosen to sample using a 0.25m2 quadrat (Figure 8) where substrate was brought to the surface for sifting through. Quantitative sampling methods were implemented with 0.25m2 quadrats positioned along transect tag lines perpendicular to the river channel. The method for the procedure followed Henley et al., (2013) where random starting points and transect distance intervals were randomly determined. Four

quadrats were placed randomly along each tagline and panned to a depth at least 15cm. Tag lines were set at least 5m apart with a sample site length not less than 25m. Once quadrats were brought to the surface, native unionids were identified, enumerated, and placed into perforated bags until timed searches were concluded. DEEP POOL HABITAT: QUALITATIVE Post quantitative search efforts were accompanied with CPUE qualitative timed search for 30 minutes with three biologists over two passes equivalent to a total of three hours. Divers began search efforts on the downstream end of the sample site and worked their way upstream side by side sweeping areas to their immediate right and left. Since depth and turbidity frequently impacted search efforts, divers worked as close to the substrate as possible in order to increase visual acuity. OTHER MESOHABITATS QUANTITATIVE Below are the standard field procedures and protocols for mussel sampling utilized in this study based on a systematic design and implemented for larger channels within the basin: (1) Measure and set up a two dimensional sample grid matrix in the stream with an area at least 10 X 5 meters divided into 50 possible sampling units (Figure 10). Figure 9. Panning with Sieves

(2) Calculate an interval distance for determining distance between quadrats and for implementing spatial integrity. Then decide how many quadrats will be sampled (at least 10 is sufficient).

3 (0,2) (2,2) 2

1 (1,0) 0 1 2 3 4 5 6 7 8 9 upstream downstream Figure 10. Matrix Grid

Matrix grid used for a Quantitative Systematic sample design. A 10 x 5 meter sample reach divided into 50 possible sampling units. As an example, three random x,y coordinates (0,2) shaded green, (1,0) shaded blue, and (2,2) shaded red will serve as starting points which continue every three meters apart (based on distance formula below).

퐿∗푊 Distance Formula, d = √ , for example, L and W are the length and width of the site, 푛/푘 n = total number of quadrats you want to be sampled, and k is the number of random starts. An example, L = 10, W = 5, and n/k = 10/3 where d equals 3.87 rounded down to three.

(2) Generate at least three random starting coordinates (x, y utilizing a website random number generator) for each random starting point, and begin sampling at these starting points and continue sampling the grid at equal intervals based on the distance formula (Figure 10). (3) Identify, enumerate, and photograph specimens and aquatic habitat types for verification. (4) Measure and record stream depth, substrate type, and three velocities (HACH FLO- MATE 2000 FH 950 in ft. /sec. directly above substrate) along the upstream side of sample grid. (5) Collect and record GPS spatial habitat perimeter coordinates using a Trimble Geo Explorer HX. Measure and record shell dimension (length in mm) using standardized calipers for shell attributes, and reveal age class structure and for detecting recent recruitment.

(6) CPUE (catch-per-unit-effort, no. /h) will be timed in man effort/hours implementing the aforementioned three pass sample option with a possible fourth pass if a listed or threatened species other than the Golden Orb is found.

This design is fairly easy to implement in the field, and provides for good spatial distribution within the matrix grid, and also efficient for rare and clustered mussel populations. The design features the use of a 0.25m2 quadrat which lies directly over the substrate, and plots are selected at regular intervals from a random starting point. Each sample unit (quadrat), that follows, lies within that same systematic sample. In this study, at least three random starting points (x, y coordinates) were generated randomly by a computer so sample variance and other demographics could be estimated without any assumptions regarding spatial distributions. For this study, at least ten quadrats were sampled at each collection site. Panning depth was at least 15cm where the firmest substrate depth terminated sample efforts. OTHER MESOHABITATS QUALITATIVE Since the westside creeks are typically very narrow and shallow, only qualitative mussel sampling (CPUE, no. /h) occurred along the West Side Creeks, while both quantitative and qualitative efforts were implemented for the lower Cibolo. Mussel search efforts for the Westside creeks were on the order of at least 0.5 h for each sample site with a search area no less than 50 m2 with two biologists. Efforts on the lower Cibolo Creek implemented both a qualitative and quantitative approach (Figure 10). After the quantitative survey was completed, a qualitative approach was implemented with a catch-per-unit-area of 150 m2. Three biologists will make three twenty minute passes over the search area. Biologists will sample directly next to each other, and sweep areas to the immediate left and right across every mesohabitat type. OTHER MEASUREMENTS Specimens from the quantitative and qualitative collections were kept separately. The length of every specimen from the quantitative sample and the first 100 individuals from the qualitative efforts were measured to document population structure before moving to the next sample site. Water quality data was also provided for both watersheds at sites within our study area to reveal water quality trends for the following parameters over a period of time : Water Quality - Dissolved Oxygen and Temperature; Total Suspended Solids (TSS), Nitrite (NO2), Nitrate (NO3), Ammonia (NH3), and Total Phosphorous (T – PO4) see Appendix A.

WESTSIDE CREEKS, TCEQ SEGMENT 1911 B, C, D, and I QUALITATIVE RESULTS Results from our qualitative search revealed very few live mussels (n=5). Only one native mussel species was collected from all four tributaries (Table 3). Thorough qualitative mussel search efforts with two SARA biologists along each creek yielded five live paper pondshell Utterbackia imbecillis. Results from Martinez Creek (W. Mulberry Street) yielded a CPUE/no.h = 10 for that particular site location, and an overall CPUE for Martinez Creek of 0.83/h. These juvenile specimens ranging from 14 – 18 mm (length only) happened to be wedged into tiny crevices within a small concrete weir. Finally, the total CPUE for all four creeks was 0.26/h reflective of the lack of mussel fauna from these tributaries. A calcified relic threeridge valve was located on San Pedro Creek (at IH-10) indicating freshwater mussels (densities unknown) did in fact inhabit one or more of the westside creeks at some time prior. Another discovery while searching Martinez Creek (Navidad Street) yielded a salt water species, variable coquina Donax variabilis likely dumped from a salt water aquarium directly into the creek. The only other live mussels were non-unionid Asian clam Corbicula spp., found in very low densities in each tributary.

Table 3. Mussel Species, Search Area m2, and CPUE's, Westside Creeks Creek Name Common Total # mussels Total Search Time CPUE, no./h Total Area searched (m2) Name (hours)

Martinez paper pondshell 5 6:00 0.83/h 740

San Pedro No Mussels 0 4:00 0 705

Apache No Mussels 0 2:40 0 325

Alazan No Mussels 0 6:20 0 992

TOTAL No Mussels 5 19 hrs. 0.26/h 2,762 m2

VELOCITIES AND DEPTHS, WESTSIDE CREEKS Generally, water visibility was fair, with narrow shallow creek beds, allowing for a thorough timed search effort. Creek depths (Table 4) were virtually the same throughout, and mean creek velocities were relatively slow with a range between -0.02 to 0.85 ft./second. Dense filamentous algae was dominant at many of the sites where low velocities occurred, often impeding the already slow velocities. Substrate components were variable across sites and creeks, but large and small gravel were dominant at 24 of 40 sites throughout (60%) and could account for the established fish communities encountered. These creeks usually pool during summer months, however, fish communities were observed at many sites with piscivorous Centrarchids such as bass and sunfish present, Poeciliids, including mollies and mosquito fish,

and other native Cyprinid types, suggesting that the fish community repopulates in early spring after the summer dry spells.

Table 4. Velocities and Depths (Mean ± SE), Westside Creeks

Abiotic Parameters Martinez Creek San Pedro Creek Alazan Creek Apache Creek Max. Velocity 0.04 0.85 0.40 0.53 (ft./sec.) Mean Velocity -0.02 ± 0.01 0.32 ± 0.10 0.10 ± 0.04 0.40 ± 0.08 (ft./sec.) Min. Velocity -0.08 0.01 -0.03 0.01 (ft./sec.) Mean Depth (ft.) 0.90 ± 0.16 0.69 ± 0.13 0.98 ± 0.16 0.74 ± 0.16

LOWER CIBOLO CREEK RESULTS, TCEQ SEGMENT 1902 Conditions during the fall of 2014, and also in the spring and summer of 2015 were very wet throughout the course of the project where flows in this area peaked on numerous occasions (Figures 11 and 12) producing record rains (USGS, 2016) throughout the region impacting sample events. High flow conditions often made it unsafe to sample and therefore sample surveys were postponed until waters receded. Two USGS site gauges were selected, one on the most upstream area of the lower Cibolo Creek and the other on the downstream end. Sampling efforts were commonly conducted above base flow conditions.

Figure 11. USGS Hydrograph Figure 12. USGS Hydrograph

Upon completion of both quantitative and qualitative surveys, a total of 81 native mussels representing 5 species were detected among the 46 sites surveyed (Tables 5 and 7) with a total search area effort at 8,250m2 along the Lower Cibolo Creek corridor. Observed natives consisted of yellow sandshell Lampsilis teres (n= 38), golden orb Quadrula aurea (n=32), paper pondshell Utterbackia imbecillis (n=3), pistolgrip Quadrula verrucosa (n=7), and louisiana fatmucket Lampsilis hydiana (n=1). Live and dead shell non-native Asian clam (Corbicula spp.) were present throughout all sample reaches. Species richness ranged from 0 - 2 per site at 0.83 ± 0.12 (mean ± SE) with the highest (2 spp.) occurring at 10 sites and the lowest (no catch) at 22 sites. Species richness in quadrat sampling versus times searches were significantly correlated applying a Spearman’s Rank Correlation (r = 0.36, P = 0.01). Comparative results for mussel presence between quantitative and qualitative sample efforts were 26% and 52%, respectively. Results for the number of species found in timed search efforts at each site was significantly higher than in quadrat sampling (0.72 ± 0.12 vs. 0.34 ± 0.09, mean ± standard error of the mean, P = 0.008, paired two-tailed t – test), indicating sufficient mussel recovery in the timed searches. See Appendix B for a table of mussel type, number and locations for both quantitative and qualitative searches. QUANTITATIVE The total number of quadrats excavated for all sites was n= 720, and number of quadrats per sample site ranged from 13 – 17 excluding the five deeper pools which had an overall larger defined search area. This range of sample quadrats was sufficient at providing good spatial coverage and estimating precise mussel densities (Strayer and Smith 2003). Only 12 of 46 sites, slightly one-fourth of the quantitative sample sites, produced measurable mussel data for determining population densities. Locating native mussels from all sample sites was somewhat challenging indicated by low mussel densities ranging from 0.0 m2 – 0.96 m2 throughout, and exhibited an overall mean and standard error at 0.152 m2 ± 0.01 which seemed to be the standard. Mussel abundance throughout the quantitative sample (Table 5) consisted of 26 observed individuals, with an estimate (population) of 312. Golden orb, a species of conservation concern, was distributed throughout much of the lower Cibolo, with the exception of sample reach FM 537 – CR 225, where only one specimen was found. There were a total of 32 golden orb specimens collected from five sample reaches 35% of the time coinciding with the low mussel density data, however, the golden orb population was distributed in very low numbers across all sample reaches.

Table 5. Quantitative Mussel Data, Lower Cibolo Creek by Sample Reaches

Sample Reach Site # Avg. CV Depth Habitat Type Mussels # Substrate Population Population (ft. sec.) (ft.) Mussels Size Density m2 (estimate)

1 0.95 1.0 run no mussels 0 gravel 0 0 3 0.76 1.6 glide no mussels 0 gravel 0 0 5 0.12 0.8 run no mussels 0 gravel 0 0 7 0.21 1.6 glide no mussels 0 gravel/sand 0 0 9 0.16 1.8 backwater no mussels 0 gravel/sand 0 0 FM 2538 -Zuehl 11 0.66 0.7 riffle no mussels 0 gravel 0 0 (reach 1) 13 0.35 1.2 run no mussels 0 gravel 0 0 15 0.90 0.8 riffle no mussels 0 gravel 0 0 17 0.05 1.7 run no mussels 0 gravel 0 0 19 0.12 1.9 glide no mussels 0 gravel 0 0

DP1 0.04 12 deep pool no mussels 0 gravel 0 0

2 1.1 0.5 riffle no mussels 0 gravel 0 0 4 0.14 1.4 pool no mussels 0 gravel 0 0 6 0.09 1.0 pool no mussels 0 gravel 0 0 8 0.14 1.8 glide golden orb 1 gravel/clay 12 0.24 Scull Crossing – CR 775 10 0.44 1.4 run yellow sandshell 2 gravel 24 0.48 (reach 2) 12 -0.01 2.4 backwater no mussels 0 silt/gravel 0 0 14 0.03 1.4 glide no mussels 0 sand/silt 0 0 DP2 0.02 13 deep pool no mussels 0 gravel 0 0 DP3 0.02 10 deep pool no mussels 0 clay/gravel 0 0 yellow sandshell, 16 -0.06 0.4 backwater 4 gravel 48 0.96 golden orb 18 1.15 1.1 riffle yellow sandshell 1 gravel 12 0.24

FM 775 – CR 337 20 0.17 1.5 run yellow sandshell 1 gravel/silt 12 0.24 (reach 3) yellow sandshell, 22 0.30 1.0 run 4 gravel 48 0.96 golden orb 24 0.48 1.6 run no mussels 0 gravel/sand 0 0 26 0.80 0.6 riffle no mussels 0 gravel 0 0

28 0.02 0.7 run no mussels 0 gravel 0 0 30 0.03 3.0 backwater no mussels 0 sand 0 0 32 0.34 2.0 run no mussels 0 gravel 0 0 FM 537 – CR 225 paper pondshell, 34 0.18 1.0 run 3 sand 36 0.72 (reach 4) golden orb 36 0.36 1.5 run yellow sandshell 1 gravel 12 0.24 38 1.04 1.2 run no mussels 0 gravel 0 0 40 0.14 1.6 run no mussels 0 sand 0 0

State Hwy. 123 – 21 0.06 1.2 glide no mussels 0 fine sand 0 0

Sample Reach Site # Avg. CV Depth Habitat Type Mussels # Substrate Population Population (ft. sec.) (ft.) Mussels Size Density m2 (estimate)

FM 2724 23 0.83 1.0 run pistolgrip 3 gravel 36 0.72 (reach 5) 25 0.61 1.1 run golden orb 4 gravel 48 0.96 27 0.41 1.0 run golden orb 1 gravel 12 0.24 29 0.61 1.5 glide no mussels 0 gravel 0 0 31 -0.01 2.5 backwater no mussels 0 clay 0 0 33 0.99 0.7 run golden orb 1 gravel 12 0.24 35 0.66 0.5 riffle no mussels 0 gravel 0 0 37 0.59 0.9 run no mussels 0 gravel 0 0 39 -0.003 1.3 backwater no mussels 0 sand/silt 0 0 41 0.47 1.8 run no mussels 0 gravel 0 0 DP4 0.53 7 deep pool no mussels 0 bedrock 0 0 DP5 0.17 8 deep pool no mussels 0 Sand/gravel 0 0 Total (n) - - - - - 26 - 312 6.24 Average Population ------0.14 Density

BIOLOGICAL STRUCTURE There are several analytical approaches which provide insight to diversity and evenness (Table 6) within a biological community. These non-parametric Indexes (Krebs 2014) are measures of heterogeneity where maximum diversity is obtained when all species abundance is equal. Usually, the critical value indicates more or less diversity and lies between 0 and 1, where a value closer to one indicates higher diversity and evenness and a value close to zero indicates low overall bio-diversity. Although sample size (n=81) was low and difficult to draw inferences from, comparisons between geospatial sample reaches and mussel diversity were made. Results for both Simpson’s tests were in the median range indicative of moderate heterogeneity within this community however, the Shannon Wiener test for Diversity was slightly over one, indicating good species diversity and also the distribution among individuals across species equally as good. Results for species diversity and evenness from all sample reaches (Table 6) except reach 1 (no specimens located) and reach 5 had scores within the moderate to good range for evenness and diversity. Reach 5 had the second highest abundance, however individuals were spread unevenly across species, and conversely reach 4 had the lowest number of individuals (n=5, 3 species) but higher scores indicating evenness among species. Several factors affecting mussel heterogeneity at sites between the upper and lower reaches could be based upon several factors such as substrate composition, mesohabitat type, or discharge rates. In this study, substrate mainly consisted of gravel and sand (35 sites) for most of the sites we encountered including reach one, and discharge rates were often above baseline flow conditions vital for mussel survival and reproduction, so at this time we have no explanation why mussels were not located there.

Figure 13. Mussels

Table 6. Indices of Evenness and Diversity by Reaches, Upstream to Downstream

Sites Sampled Mussels Species Simpson’s Index of Simpson’s Shannon-Wiener Found Richness Diversity (1-D) Evenness Diversity Index (ln)

46 (including deep pools) 81 5 0.61 0.52 1.01

Reach 1 0 0 0 0 0 (FM 2538 – Zuehl)

Reach 2 30 4 0.39 0.64 0.70 (Scull Crossing – F775)

Reach 3 19 3 0.44 0.60 0.80 (FM 775 – CR 337)

Reach 4 5 3 0.64 0.93 1.04 (FM 537 – CR 225)

Reach 5 27 2 0.35 0.77 0.52 (CR 389 – FM 2724)

QUALITATIVE Qualitative tactile and visual approaches included substrate excavation (panned sediment) down at least 15cm or until firm. Panning was not a constant method, but one used with discretion and dependent on ambient water conditions such as depth and turbidity. During these search efforts (Table 7), the objective was to cover as much area as possible. Under these conditions, panned scoops of sediment were brought to the surface and inspected for mussels using mussel sieves. The qualitative sample design, which accounted for three passes, produced an overall CPUE ranging from 0 – 2.33/h. The mean CPUE for the mussel community across all sample sites was 0.40 ± 0.08 mussel/h (mean ± SE), while the mean CPUE for species of conservation concern (golden orb) was 0.13 ± 0.05/h. The number of mussels from qualitative samples was at 55 individuals and higher than our quantitative sediment samples. Overall, 31 sites had CPUE’s over 0.333 and timed efforts were greater than mussels per meters

squared. Mussels were found at 52% of sites, and golden orb were located at 35% of sites, suggesting mussel populations are distributed throughout the watershed, but in very low densities.

Table 7. Qualitative Mussel Data, Lower Cibolo Creek

Site # Avg. # of CPUE CV Depth Habitat Type Mussels Substrate Mussels (no./h) Sample Reach (ft. /sec.) (ft.) 1 0.95 1.0 run gravel no mussels 0 0 3 0.76 1.6 glide gravel no mussels 0 0 5 0.12 0.8 run gravel no mussels 0 0 7 0.21 1.6 glide gravel/sand no mussels 0 0 9 0.16 1.8 backwater gravel/sand no mussels 0 0 FM 2538 – Zuehl 11 0.66 0.7 riffle gravel no mussels 0 0 (reach 1) 13 0.35 1.2 run gravel no mussels 0 0 15 0.90 0.8 riffle gravel no mussels 0 0 17 0.05 1.7 run gravel no mussels 0 0 19 0.12 1.9 glide gravel no mussels 0 0 DP1 0.04 12 deep pool gravel no mussels 0 0 2 1.1 0.5 riffle gravel yellow sandshell 1 0.3333 4 0.14 1.4 pool gravel yellow sandshell 3 1 6 0.09 1.0 golden orb 1 0.3333 pool gravel yellow sandshell 1 0.3333 8 0.14 1.8 golden orb 2 0.6666 glide gravel/clay yellow sandshell 1 0.3333 10 0.44 1.4 golden orb 1 0.3333 Scull Crossing – FM775 run gravel (reach 2) yellow sandshell 2 0.6666 12 -0.01 2.4 louisiana fatmucket 1 0.3333 backwater silt/gravel yellow sandshell 6 2 14 0.03 1.4 glide sand/silt yellow sandshell 6 2 DP2 0.02 13 deep pool gravel no mussels 0 0 DP3 0.02 10 yellow sandshell 1 0.3333 deep pool clay/gravel paper pondshell 1 0.3333 16 -0.06 0.4 backwater gravel yellow sandshell 2 0.6666 18 1.15 1.1 riffle gravel yellow sandshell 1 0.3333 yellow sandshell 1 0.3333 FM 775 – CR 337 20 0.17 1.5 run gravel/silt (reach 3) pistolgrip 1 0.3333 22 0.30 1.0 run gravel yellow sandshell 2 0.6666 24 0.48 1.6 run grave/sand golden orb 1 0.3333 26 0.80 0.6 riffle gravel golden orb 1 0.3333 28 0.02 0.7 run gravel no mussels 0 0 30 0.03 3.0 backwater sand no mussels 0 0 32 0.34 2.0 run gravel golden orb 1 0.3333 FM 537 – CR225 34 0.18 0.9 run sand no mussels 0 0 (reach 4) 36 0.36 1.5 run gravel no mussels 0 0 38 1.04 1.2 run gravel no mussels 0 0 40 0.14 1.6 pool sand no mussels 0 0 21 0.06 1.2 glide fine sand no mussels 0 0 23 0.83 1.0 run gravel golden orb 3 1 golden orb 1 0.3333 25 0.61 1.1 run gravel pistolgrip 2 0.6666 27 0.41 1.0 run gravel golden orb 1 0.3333 29 0.61 1.5 glide gravel no mussels 0 0 State Hwy. 123 – 31 -0.01 2.5 backwater clay no mussels 0 0 FM 2724 33 0.99 0.7 run gravel golden orb 1 0.3333 (reach 5) 35 0.66 0.5 riffle gravel golden orb 1 0.3333 37 0.59 0.9 run gravel pistolgrip 1 0.3333 39 -0.003 1.3 backwater sand/silt golden orb 5 1.6666 41 0.47 1.8 run gravel golden orb 1 0.3333 DP4 0.53 7.0 deep pool bedrock no mussels 0 0 golden orb 1 0.3333 DP5 0.17 8.0 deep pool sand/gravel yellow sandshell 1 0.3333 Total (n) - - - - 55 18.3322 Average CPUE - - - - - 0.40

Five deeper pool sample sites from the lower Cibolo Creek (Table 8) were included in this study to document mussel presence/absence in deeper water. Surprisingly, quadrat sediment samples revealed quantitative mussel densities of zero from every quadrat at every single site, however, timed CPUE searches efforts produced a total of four mussels consisting of two yellow sandshell, one golden orb, and one paper pondshell from two separate pools. Substrate composition consisted of various components of smaller gravel, sand, silt, mud, and larger cobble making for suitable mussel habitat and the likelihood for detectability. Asian clam, Corbicula spp., were at times present but in low densities.

Table 8. Deep Pooled Mussel Sample Sites and Results

Lower Cibolo Qualitative Sampling Quantitative Sampling Creek Pool # Location Coordinates Reach Area Species CPUE No. of No. of Density m2 (L x W, m2) (no./h) transects quadrats 1 FM 2538 29.45148 30 x 15 = 450 No Mussels 0 6 24 0 -98.12305 2 Scull Crossing 29.39048 25 x 20 = 500 No Mussels 0 5 20 0 -98.12568 3 Scull Crossing 29.38952 25 x 17 = 425 yellow sandshell 0.6666 5 20 0 -98.12447 (1), paper pondshell (1) 4 SH 123 29.01452 25 x 12 = 300 No mussels 0 5 20 0 -97.92937 5 CR 389 29.01484 25 x 17 = 425 yellow sandshell 0.6666 5 20 0 -97.91898 (1), golden orb (1)

VELOCITIES AND DEPTHS Abiotic parameters such as stream velocity and depth are critical components for instream aquatic habitats and the fauna that it supports. Instream flow is water moving in a defined area that provides habitats for the flora and fauna within a stream channel where it is also inclusive of the adjacent riparian zone that acts as a natural buffer protecting the stream from various pollutants and high sediment loads. Instream flows are basically the volume of flowing water required to sustain a healthy aquatic ecosystem based on that quantity and the timing of flows (seasonal) that can adequately support life. Currently, statewide sampling efforts by the TPWD across all major basins are collecting as much data as possible to determine these flows. In this study, three velocities (ft. /sec.) and one depth (ft.) were measured and collected (Table 9) at each sample site directly above the substrate on both edges and in the center of each mesohabitat type on the upstream side perpendicular to the channel. During the spring of 2015, the central Texas region including the lower Cibolo Creek received an unusual amount of precipitation on a regular basis. Subsequently, more often than not, higher flow conditions were encountered most of the time. Flash flooding was often the case where the Cibolo would flash, drain, and flash again over already saturated soils allowing for even faster runoff. Substrate composition is a vital and integral part of mussel survival and sustainability. Their ability to maneuver sediment and dig in allows their hydrodynamic shapes to create less drag.

Dependent upon species and substrate type, whether soft or firm, most will attempt to etch a crevice and dig in. In this study, substrate components in the lower Cibolo were comprised of various mixtures of small to large, and fine to gritty types. Overall, copious amounts of small and large gravel associated with sand dominated all the sites where this mixture was observed at 40 sites and prevalent 87% of the time. Other substrate types such as fine silt, soft or hard clays, and bedrock were present but not as dominant at the particular sites.

Table 9. Velocities and Depths (mean ± SE), Lower Cibolo Creek ABIOTIC Mesohabitat Mesohabitat Mesohabitat Mesohabitat PARAMETER Glide (7), Run (19) Riffle (6) Pool (8) Backwater (6) Max. Velocity (ft. /sec.) 1.04 1.15 0.66 0.16

Mean Velocity (ft. /sec.) 0.41 ± 0.01 0.87 ± 0.40 0.18 ± 0.05 0.02 ± 0.03

Min. Velocity (ft. /sec.) 0.02 0.66 0.01 -0.003

Mean Depth (ft.) 1.3 ± 0.08 0.82 ± 0.94 6.6 ± 1.8 1.9 ± 0.03

POPULATION STRUCTURE All live native mussels collected were measured from anterior to posterior (length only) using slide calipers. Justification for more quantifiable results would require a much larger sample size, therefore making inferences regarding size classes should be used with caution. Furthermore, this data should be utilized only as a guideline and not for making assumptions intended for any design based inference model. Mussel shell dimensions (inflated or compressed eco-phenotypes) maybe subjective to geospatial location, whether they are found upstream in faster flowing water headwaters, or downstream located in less turbulent currents associated with more laminar flows. Also, intra-specific mussel dimensions can be misleading, especially when species specific inherent genetic traits exhibit various individual shapes and sizes within a population. Mussel data collected between timed CPUE and quadrat sampling were analyzed across 46 sites from the lower Cibolo Creek were sorted into four size classes as adopted from Johnson et al. (2012):

 0-30mm  31-40mm  41-69mm  70-140mm The presence of the smaller class sizes for yellow sandshell, golden orb and paper pondshell indicate that recruitment is occurring. The lack of the smaller classes for the pistolgrip and the louisiana fatmucket may indicate a lack of recruitment, or it may simply be due to the small number of pistolgrips (n=7) and louisiana fatmucket (n=1) found. Figures 14-18 show the class structure of each mussel found.

The limited data set indicates that results revealed qualitative timed searches were effective and representative of larger specimens from the larger classes of yellow sandshell, Lampsilis teres where the two largest classes were represented by 13 and 14 individuals, respectively. Based on this dataset, all four other species were either under represented or in very low numbers where paper pondshell Utterbackia imbecillis (n = 0), golden orb Quadrula aurea (n= 0), louisiana fatmucket Lampsilis hydiana (n = 0), and pistolgrip Quadrula verrucosa (n = 2) may be indicative of a lack of larger sexually reproducing adults. Results from the timed qualitative searches for smaller classes of recent recruitment were under represented by pistolgrip (n = 0), louisiana fatmucket (n = 0), yellow sandshell (n = 2), and paper pondshell (n = 1). Golden orb was the only species representative in the 0 to 30 mm and the 31 to 40 mm classes (n = 14) and might be a cause for concern for the lack of overall recent recruitment. A possible and likely explanation could be that visual tactile searches were impacted by water clarity (turbidity) and depth and the likelihood of detecting larger more sculptured specimens protruding just above or immediately below the substrate under these conditions is greater. This seems to be a logical scenario for the success in locating larger specimens during timed searches. All small to medium size specimens (41-69mm) across all species were represented in low numbers or not represented at all except yellow sandshell (n = 14) suggesting that medium sized individuals were difficult to locate. This study revealed that for the most part, timed qualitative searches are an effective, useful, and necessary tool in estimating mussel abundance, but can overestimate for larger species and simultaneously underestimate smaller buried individuals. Quadrat searches are designed to detect both small (<30mm) and large (70-140mm) individuals and unbiased to size or shell length, however in this study, smaller individuals were completely not detected from pistolgrip and louisiana fatmucket. Results also revealed that younger size classes were under represented by all species except golden orb which had the highest number of medium size specimens (n=14, 31-40mm), and to a much lesser extent, paper pondshell and yellow sandshell. Specimens of louisiana fatmucket, paper pondshell, and golden orb, were absent in the largest size class. Quadrat searches revealed moderate numbers of yellow sandshell and golden orb from the medium size class (41-69mm) but lacked among pistolgrip, louisiana fatmucket, and paper pondshell. Quantitative results were representative across all species in different size classes except for louisiana fatmucket and the lack of smaller pistolgrip and louisiana fatmucket is difficult to explain since quadrat mesh was implemented to minimize for sample error. A plausible explanation could simply be a lack of larval recruitment in these groups which may be a cause for concern. Random quadrat sampling provides for more precise mussel densities and size class structure that is pertinent and vital for gaging the integrity of mussel survival. It can also reveal insight to issues within a population regarding size representation.

pistolgrip size classes (n=7) Timed Quadrat 4

Number of 2 mussels 1

0 0-30mm 31-40mm 41-69mm 70-140mm

Figure 14. Pistolgrip Size Classes

yellow sandshell size classes (n=38) Timed Quadrat 15

Number 10 of mussels 5

0 0-30mm 31-40mm 41-69mm 70-140mm

Figure 15. Yellow Sandshell Size Classes

golden orb size classes (n=32) Timed Quadrat 15

10 Number of mussels 5

0 0-30mm 31-40mm 41-69mm 70-140mm Figure 16. Golden Orb Size Classes

louisiana fatmucket size classes (n=1) Timed Quadrat 1.2

Number 0.8 of mussels 0.6

0.4

0.2

0 0-30mm 31-40mm 41-69mm 70-140mm

Figure 17. Louisiana Fatmucket Size Classes

paper pondshell size classes (n=3) Timed Quadrat 2.5

1.5 Number of mussels 1

0.5

0 0-30mm 31-40mm 41-69mm 70-140mm

Figure 18. Paper Pondshell Size Classes

ASSOCIATED MUSSEL MESOHABITATS Mussel habitat is an ongoing topic in the scientific mussel community that still requires more research for predicting mussel presence in lotic ecosystems. Aquatic mesohabitats can be characterized by either slow or fast moving water with soft or firm substrates derived and driven by variables such as streambed gradient and current velocities within a stream channel. Mussel species can occur across a variety of mesohabitats where flowing water with higher dissolved oxygen is often associated with more biodiversity, and conversely pooled water that may support more tolerant mussel species. This study classified four separate mesohabitat (Figure 19) types based on eighty-one individuals. Four mesohabitats were assessed for association by species across pools, backwater, riffles, and glide/run combinations. Glides and runs were utilized the most, however these mesohabitats also dominated others by more than two times and results will be somewhat biased. Relative percentages showed yellow sandshell (25%), and golden orb (26%) and to a much lesser extent pistolgrip (9%) associated to runs and or glides where current velocities are not as swift as riffles and within glides more laminar. Both species (yellow sandshell, golden orb) also seemed to be found in backwater areas usually associated with slack and or standing water. Although percentages were much lower for the two in backwater areas, this might provide some insight about the life cycles of both, first, are both generalists or specialists types and second, are both species able to survive in various current velocities whether still or moderate and within substrates consisting of various fine or larger heterogeneous particles. Another question arises, does either species require flowing water during the spawning season? This study could provide evidence that both species are more generalist types and can survive and successfully reproduce within these mesohabitat types indicative of their adaptability.

One concern that was observed was the lack of individuals located in riffling water. These habitats are generally associated with swift currents and higher dissolved oxygen. Once again, golden orb and yellow Sandshell were the only two species associated to riffle mesohabitats. Possible explanations are larger fishing birds, feral hogs, or small mammals that frequent the banks and seize the opportunity to wade in and search the shallower areas within these habitat types.

Habitat Utilization, n=81 30%

25%

20% Relative Percent 15% 10%

0% yellow ss golden orb paper pondshell pistolgrip louisiana fatmucket

Glide/Run (26) Pool (8) Backwater (6) Riffle (6)

Figure 19. Mussel Mesohabitats Relative to Mussel Abundance

GEOSPATIAL CONTRAST The farthest upstream reach began at FM 2538 (Figure 20) near New Berlin, Texas where no mussels were located. Substrate texture at these sample sites were certainly adequate to support mussel fauna where mixtures consisted of small and larger gravel and sand. Possible explanations could include fish and host specificity, sample error, or a water quality issue from local waste water treatment plants. Moving downstream, the next reach (Scull Crossing) was upstream to the City of LaVernia, TX. where the highest number of mussels were collected (n=30), and subsequently followed by the reach from FM 775 to CR 337 lying east of LaVernia and just outside of the city where Cibolo Creek drains. Here the number of mussels between FM 775 to CR 337 (n=19) dropped slightly from the previous reach and were almost non- existent (n=5) between FM 537 and CR 225. There is a definite disconnection between mussel abundance across reaches from the middle area to the farthest downstream. Once again, mesohabitat integrity and substrate composition did not seem to be an issue for mussel

presence and thorough search efforts resulted in just a few individuals. The last sample reach was near and just above the community of Panna Maria, TX. where mussel abundance rose to 27 individuals and substrate composition was consistent with all previous reaches and sample sites. Local waste water treatment plants near the towns of Marion, TX. Stockdale, TX. and LaVernia, TX. are all permitted by the TCEQ to discharge treated waste water. Finally, locating multi-species groups (mussel beds) in riverine systems is typically a hit or miss scenario due to

Relative Spatial Reaches, n=81 35 30 25 Number of 20 mussels 15 10 5 0 FM 2538 - Zuehl Scull Crossing - FM FM 775- CR 337 FM 537-CR 225 CR389-FM2724 Crossing 775 Farthest Upstream Middle Farthest Downstream

Figure 20. Relative Spatial Reaches on Mussel Abundance

their patchy and clustered distributions. It is a fact that mussels can be located in the most unexpected places and, conversely, not found where expected. The sample method design for this study was based on randomness hoping to locate clusters, but in the process actually discovered some disjoint between sample reaches and mussel abundance from several reaches.

LITERATURE CITED

Bogan, A. E. 1993. Freshwater bivalve extinctions (Mollusca: Unionoida): A search for causes. Am. Zool. 33:599–609. Burlakova, L. E., A. Y. Karatayev, V. A. Karatayev, M. E. May, D. L. Bennett, and M. J. Cook. 2011. Biogeography and conservation of freshwater mussels (Bivalvia: Unionidae) in Texas: patterns of diversity and threats. Diversity and Distributions, 17: 393–407. Fuller, S. L. H. 1974. Clams and mussels (Mollusca: Bivalvia). Pollution ecology of freshwater invertebrates. Academic Press, New York Green, R. H., R. C. Bailey, S. G. Hinch, J. L. Metcalfe, and V. H. Young. 1989. Use of Freshwater Mussels, (Bivalvia: Unionidae) to monitor the near shore environment of lakes, J great lakes Res., 15, 635-644.

Henley, W. F., M. J. Pinder, B. T. Watson, and R. Neves. 2013. Status of Freshwater Mussels in the Middle Fork Holston River, Virginia. 2013. Walkerana: The Journal of the Freshwater Mollusk Conservation Society 16:68-80. Howells, R.G. 1995. Distributional surveys of freshwater bivalves in Texas: progress report for 1993. Texas Parks and Wildlife Department, management Data Series 119, Austin.46pp. Howells, R. G., R. W. Neck, and H. D. Murray. 1996. Freshwater Mussels of Texas, Texas. Parks and Wildlife Press, Austin, Texas. Howells, R. G. 1997a. Distributional surveys of freshwater bivalves in Texas: progress report for 1996. Texas Parks and Wildlife Department, Data Management Series 144,Austin.52pp. Johnson, M.S., W. F. Henley, R. J. Neves, J. W. Jones, R. S. Butler, and S. D. Hanlon. 2012. Freshwater mussels of the Powell River, Virginia and Tennessee: abundance and distribution in a biodiversity hotspot. Walkerana, Journal of the Freshwater Mollusk Conservation Society 15, 83–98. Karatayev A. Y. and L. E. Burlakova. 2008. Interagency Report to the Texas Water Development Board. Final Report. Distributional Survey and Habitat Utilization of Freshwater Mussels. Krebs, C. J. 2014. Ecological Methodology, 6th ed., Addison-Wesley Educational Publishers, Inc., Menlo Park, CA. Metcalfe J. L. and M. N. Charlton. 1990. Freshwater Mussels as biomonitors for organic industrial contaminants and pesticides in the St. Lawrence River. Science of the Total Environment, 97-8. 595-615. Randklev, Charles 2014. Personal communication, Texas A and M Agrilife Extension, Freshwater Mussel Symposium, Kerrville, TX. Richter, B. D., R. Mathews, D. L. Harrison, and R. Wigington. 2003. Ecologically sustainable water management: managing river flows for ecological integrity. Ecological Applications, 13(1): 206-224. Robertson, Clint 2014. Personal communication, Texas Parks and Wildlife Department, Meadows River Study Center, San Marcos, TX. SARA, 2008. Texas Clean Rivers Program, San Antonio River Basin Summary Report. SARA, 2011. Instream Flow Study of the Lower San Antonio River and Lower Cibolo Creek Interim Progress Report and Instream Flow Recommendations. 101pp. Strahler, A. N. 1952. "Hypsometric (area-altitude) analysis of erosional topology", Geological Society of America Bulletin 63 (11): 1117–1142. TCEQ, 2014. Texas Commission on Environmental Quality-2014 Texas Integrated Report for the Clean Water Act Sections 305(b) and 303(d). www.tceq.texas.gov/waterquality/assessment/14twqi/14txir Strayer, D. L., and D. R. Smith. 2003. A Guide to sampling freshwater Mussel Populations. American Fisheries Society, Monograph 8, Bethesda, Maryland. Turgeon, D.D. Jr., J. F. Quinn, A. E. Bogan, E. V. Coan, W. K. Emerson, W. G. Lyons, W. L. Pratt. G. E. Roper, A. Scheltma, F. G. Thompson and J. D. Williams. 1988. Common and scientific names of aquatic invertebrates from the United States and Canada: mollusks, 2nd edition. American Fisheries Society, Bethesda, Maryland, 277 pp. USGS, (2016). USGS Current Water Data for the Nation, USGS Science for a changing world. [Online]. Available: http://waterdata.usgs.gov/nwis/rt, [2016].

Vaughn, C.C., C.M. Taylor, K. J. Eberhard. 1997. A comparison of the effectiveness of timed searches vs. quadrat sampling in mussel surveys. Pages 157 – 162 in K. S. Cummings, A. C. Buchanan, C. A. Mayer, and T. J. Naimo, editors. Conservation and management of freshwater mussels II. Initiatives for the future. Upper Mississippi River Conservation Committee, Rock Island, Illinois. Vaughn, C.C. and C. Taylor. 1999 Impoundments and the Decline of Freshwater Mussels; a Case Study of an Extinction Gradient. Conservation Biology 13:912-920. Williams, J. D., M. L. Warren, Jr. K. S. Cummings, J. L. Harris and R. J. Neves. 1993. Conservation of freshwater mussels of the United States and Canada. Fisheries 18:6–22.

APPENDIX A: WATER QUALITY AT SELECTED SITES.

Table 10. Impairment and concerns from 2014 Integrated Report.

Creeks/Segment E. Fish D.O. D.O. Habitat Nitrate Total Ammonia coli (average (min. Phosphorus grab) grab) Martinez (1911) NS NA CS FS NA NC NC NC San Pedro NS NS NC FS CS CS CS NC (1911) Alazan (1911) NS NA NC FS NA NC NC CS Apache (1911) NS NA CS FS NA CS NC NC Lower Cibolo NS CN NC FS NC NC NC NC (1902) ** FS = fully supporting, NA = not assessed, NS = not supporting, NC = no concern, CS = concern for water quality based on screening level status, CN=Use Concern.

WESTSIDE CREEKS, WATER QUALITY DATA, TCEQ SEGMENT 1911. Standards for Conventional Water Quality Parameters and Nutrient Screening Levels. a) Total Suspended Solids (TSS): No State Standard set. b) Total Phosphorus: Screening Level – 0.69 (mg/L) c) Temperature Standard – 90 degrees Fahrenheit (32.2 degrees Celsius) d) Dissolved Oxygen Standard – 3.0 mg/L (minimum grab) San Pedro and Apache Creeks; 2.0 mg/L (minimum grab) for Martinez and Alazan Creeks. e) Nitrate Nitrogen: Screening Level – 1.95 mg/L, f) Ammonia Nitrogen: Screening Level – 0.33 mg/L.

APACHE CREEK AT BRAZOS: Date Range of Analysis (2009-2014) TSS – No state standard. Two extremely high pulses for 2009 and 2010. Total Phosphorus –Data values were below screening level. Water Temperatures and Dissolved Oxygen – High temperatures coincide with seasonality and low dissolved oxygen levels occurred during the summers of 2009, 2011, and 2014 where these narrow creeks become abundant with filamentous algae impacting dissolved oxygen levels during non-photosynthetic periods. Nitrate and Ammonia – Ammonia levels were low and maintained below screening levels, however, high Nitrate levels were constant throughout suggesting possible influx of nutrients typical of inner city streams and creeks (likely Eutrophic).

Temperature Apache Creek at Brazos Dissolved Oxygen 35

15 mg/L mg/L Celsius &

0 Apache Creek at Brazos Total Phosphorus

0.14

1/12/2009 4/12/2009 7/12/2009 1/12/2010 4/12/2010 7/12/2010 1/12/2011 4/12/2011 7/12/2011 1/12/2012 4/12/2012 7/12/2012 1/12/2013 4/12/2013 7/12/2013 1/12/2014 4/12/2014 7/12/2014

10/12/2010 10/12/2011 10/12/2012 10/12/2013 0.12 10/12/2009 0.1 0.08 mg/L 0.06 0.04 0.02 Nitrite (NO2) Nitrate (NO3) 0 Apache Creek at Brazos NH3 (non-distilled) 6

4/26/2011 1/26/2009 4/26/2009 7/26/2009 1/26/2010 4/26/2010 7/26/2010 1/26/2011 7/26/2011 1/26/2012 4/26/2012 7/26/2012 1/26/2013 4/26/2013 7/26/2013 1/26/2014 4/26/2014 7/26/2014

10/26/2011 10/26/2009 10/26/2010 10/26/2012 10/26/2013

4 mg/L

1/26/2012 1/26/2009 4/26/2009 7/26/2009 1/26/2010 4/26/2010 7/26/2010 1/26/2011 4/26/2011 7/26/2011 4/26/2012 7/26/2012 1/26/2013 4/26/2013 7/26/2013 1/26/2014 4/26/2014 7/26/2014

10/26/2009 10/26/2010 10/26/2011 10/26/2012 10/26/2013

MARTINEZ CREEK AT RUIZ: Date Range of Analysis (2009-2014) TSS: No state standard. One high pulse; where peak dates are synonymous to Apache Creek. Total Phosphorus: Data values were below screening level. Water Temperature and Dissolved Oxygen: Low dissolved oxygen levels relative to high water temperatures were constant from late spring until the end of summer throughout the date range where dissolved oxygen was its lowest during August of 2009. Nitrate and Ammonia: Nitrate levels were under the screening level, however one ammonia value was 1.02 mg/L in January 2011.

Martinez Creek at Ruiz Str. TSS 70

40 mg/L 30

Total Phosphorus

1/1/2009 3/1/2009 5/1/2009 7/1/2009 9/1/2009 1/1/2010 3/1/2010 5/1/2010 7/1/2010 9/1/2010 1/1/2011 3/1/2011 5/1/2011 7/1/2011 9/1/2011 1/1/2012 3/1/2012 5/1/2012 7/1/2012 9/1/2012 1/1/2013 3/1/2013 5/1/2013 7/1/2013 9/1/2013 1/1/2014 3/1/2014 5/1/2014 7/1/2014

11/1/2010 11/1/2011 11/1/2012 11/1/2013 11/1/2009 Martinez Creek at Ruiz Str. 0.18 0.16 0.14 0.12 0.1

mg/L 0.08 0.06 0.04 0.02

11/1/2013 11/1/2009 11/1/2010 11/1/2011 11/1/2012

Temperature Martinez Creek at Ruiz Str. Dissloved Oxygen 35

15 mg/L mg/L Celsius &

1/8/2009 4/8/2011 7/8/2011 4/8/2009 7/8/2009 1/8/2010 4/8/2010 7/8/2010 1/8/2011 1/8/2012 4/8/2012 7/8/2012 1/8/2013 4/8/2013 7/8/2013 1/8/2014 4/8/2014 7/8/2014

10/8/2009 10/8/2010 10/8/2011 10/8/2012 10/8/2013

Nitrite (NO2) Martinez Creek at Ruiz Str. Nitrate (NO3) NH3 (non-distilled) 1.2

0.8

0.6 mg/L

0.4

0.2

9/1/2009 7/1/2010 5/1/2011 3/1/2012 1/1/2013 1/1/2009 3/1/2009 5/1/2009 7/1/2009 1/1/2010 3/1/2010 5/1/2010 9/1/2010 1/1/2011 3/1/2011 7/1/2011 9/1/2011 1/1/2012 5/1/2012 7/1/2012 9/1/2012 3/1/2013 5/1/2013 7/1/2013 9/1/2013 1/1/2014 3/1/2014 5/1/2014 7/1/2014

11/1/2013 11/1/2009 11/1/2010 11/1/2011 11/1/2012

ALAZAN CREEK AT TAMPICO: Date Range of Analysis (2009-2014) TSS: No state standard. Two moderate pulses contributed during March 2010 and May 2011, however creek did not seem prone to post precipitous high sediment loads. Total Phosphorus: Data values were below screening levels. Water Temperature and Dissolved oxygen: Water temperatures remained high during early spring and during summer months annually. Dissolved Oxygen remained above the minimum threshold throughout date range and was not an issue. Nitrate and Ammonia: Nitrate levels were below screening thresholds throughout the date range, but ammonia levels surpassed the screening level on several occasions.

TSS Alazan Creek at Tampico 25

15 mg/L 10

3/1/2012 7/1/2012 1/1/2009 3/1/2009 5/1/2009 7/1/2009 9/1/2009 1/1/2010 3/1/2010 5/1/2010 7/1/2010 9/1/2010 1/1/2011 3/1/2011 5/1/2011 7/1/2011 9/1/2011 1/1/2012 5/1/2012 9/1/2012 1/1/2013 3/1/2013 5/1/2013 7/1/2013 9/1/2013 1/1/2014 3/1/2014 5/1/2014 7/1/2014

11/1/2012 11/1/2009 11/1/2010 11/1/2011 11/1/2013

Alazan Creek at Tampico Total Phosphorus

mg/L

11/1/2010 11/1/2009 11/1/2011 11/1/2012 11/1/2013

Temperature Alazan Creek at Tampico Dissolved Oxygen 35

15 mg/L mg/L Celsius &

1/8/2009 4/8/2009 7/8/2009 1/8/2010 4/8/2010 7/8/2010 1/8/2011 4/8/2011 7/8/2011 1/8/2012 4/8/2012 7/8/2012 1/8/2013 4/8/2013 7/8/2013 1/8/2014 4/8/2014 7/8/2014

10/8/2009 10/8/2010 10/8/2011 10/8/2012 10/8/2013

Nitrite (NO2) Nitrate (NO3) Alazan Creek at Tampico NH3 (non-distilled) 0.9

0.8

0.7

0.6

mg/L 0.5

0.4

0.3

0.2

0.1

11/1/2009 11/1/2010 11/1/2011 11/1/2012 11/1/2013

SAN PEDRO CREEK AT MITCHELL: Date Range of Analysis: (2012-2014) TSS: No state standard. One extreme flood event where flows exceeded 80,000 cfs in downtown San Antonio, TX area. Suspended solids were elevated for a brief period after event. Total Phosphorus: Data values were below screening levels throughout date range. Water Temperature and Dissolved Oxygen: Limited data showed that water temperature peaked in July of 2014 and dissolved oxygen levels remained constant throughout able to support aquatic life. Nitrate and Ammonia: Ammonia levels remained very low throughout the date range in San Pedro Creek but higher levels of Nitrate exceeded the screening level multiple times.

San Pedro Creek at Mitchell TSS 350 300 250 200

mg/L 150 100 50

4/5/2013 6/5/2014 1/5/2013 2/5/2013 3/5/2013 5/5/2013 6/5/2013 7/5/2013 8/5/2013 9/5/2013 1/5/2014 2/5/2014 3/5/2014 4/5/2014 5/5/2014 7/5/2014 8/5/2014

12/5/2012 10/5/2013 11/5/2013 12/5/2013

Total San Pedro Creek at Mitchell Phosphorus 0.45 0.4 0.35 0.3 0.25

mg/L 0.2 0.15 0.1 0.05

9/5/2013 1/5/2013 2/5/2013 3/5/2013 4/5/2013 5/5/2013 6/5/2013 7/5/2013 8/5/2013 1/5/2014 2/5/2014 3/5/2014 4/5/2014 5/5/2014 6/5/2014 7/5/2014 8/5/2014

12/5/2012 10/5/2013 11/5/2013 12/5/2013

Temperature

San Pedro Creek at Mitchell Dissolved Oxygen 40

20 mg/L mg/L Celsius & 15

1/5/2013 2/5/2013 3/5/2013 4/5/2013 5/5/2013 6/5/2013 7/5/2013 8/5/2013 9/5/2013 1/5/2014 2/5/2014 3/5/2014 4/5/2014 5/5/2014 6/5/2014 7/5/2014 8/5/2014

12/5/2012 10/5/2013 11/5/2013 12/5/2013

Nitrite (NO2)

Nitrate (NO3)

San Pedro Creek at Mitchell Ammonia (non-distilled) 4

3.5

2.5 mg/L

1.5

0.5

8/5/2013 1/5/2013 2/5/2013 3/5/2013 4/5/2013 5/5/2013 6/5/2013 7/5/2013 9/5/2013 1/5/2014 2/5/2014 3/5/2014 4/5/2014 5/5/2014 6/5/2014 7/5/2014 8/5/2014

12/5/2012 10/5/2013 11/5/2013 12/5/2013

LOWER CIBOLO CREEK, WATER QUALITY DATA, TCEQ SEGMENT 1902. Standards for Conventional Water Quality Parameters and Nutrient Screening Levels. a) Total Suspended Solids (TSS): No State Standard b) Total Phosphorus: Screening Level – 0.69 (mg/L) c) Temperature Standard – 90 degrees Fahrenheit (32.2 degrees Celsius) d) Dissolved Oxygen Standard – 3.0 mg/L (minimum grab) e) Nitrate Nitrogen: Screening Level – 1.95 mg/L f) Ammonia Nitrogen: Screening Level – 0.33 mg/L.

CIBOLO CREEK AT CR 389: Date Range of Analysis: (2009 - 2014) TSS: No State Standard. Three minor and two major pulses within a five year period. Total Phosphorus: Data values were consistently below screening levels except for one period during July 2013 were level was slightly above the screening criteria. Water Temperature and Dissolved Oxygen: Data values were all well within standards for both parameters levels. Nitrate and Ammonia: Data collected indicated that nitrate met the screening criteria however, in the 2014 Integrated report, the TCEQ has identified upstream of this reach as a concern for nitrate nitrogen. Ammonia levels were below screening criteria.

Cibolo Creek at CR 389 TSS 400

350

300

250

200 mg/L 150

100

5/3/14 2/3/09 5/3/09 8/3/09 2/3/10 5/3/10 8/3/10 2/3/11 5/3/11 8/3/11 2/3/12 5/3/12 8/3/12 2/3/13 5/3/13 8/3/13 2/3/14 8/3/14

11/3/09 11/3/10 11/3/11 11/3/12 11/3/13

Cibolo Creek at CR 389 Total Phosphorus 0.9 0.8 0.7 0.6 0.5

0.4 mg/L 0.3 0.2 0.1

5/3/12 2/3/09 5/3/09 8/3/09 2/3/10 5/3/10 8/3/10 2/3/11 5/3/11 8/3/11 2/3/12 8/3/12 2/3/13 5/3/13 8/3/13 2/3/14 5/3/14 8/3/14

11/3/12 11/3/09 11/3/10 11/3/11 11/3/13

Temperature Cibolo Creek at CR 389 Dissolved Oxygen 35

15 mg/L mg/L Celsius & 10

5/3/09 2/3/12 2/3/09 8/3/09 2/3/10 5/3/10 8/3/10 2/3/11 5/3/11 8/3/11 5/3/12 8/3/12 2/3/13 5/3/13 8/3/13 2/3/14 5/3/14 8/3/14

11/3/09 11/3/10 11/3/11 11/3/12 11/3/13

Nitrite (NO2) Cibolo Creek at CR 389 Nitrate (NO3) Ammonia (NH3) 3

2.5

1.5 mg/L

0.5

5/3/13 2/3/09 5/3/09 8/3/09 2/3/10 5/3/10 8/3/10 2/3/11 5/3/11 8/3/11 2/3/12 5/3/12 8/3/12 2/3/13 8/3/13 2/3/14 5/3/14 8/3/14

11/3/09 11/3/10 11/3/11 11/3/12 11/3/13

APPENDIX B: LOCATION, NUMBER AND TYPES OF MUSSELS FOUND.

Quantitative Qualitative Common Reach Site # Latitude Longitude No. of No. of Total Name mussels mussels

1 29.4531 -98.12437 no mussels 0 0 0 no mussel 3 29.4505 -98.12219 0 0 0 s 5 29.45027 -98.1213 no mussels 0 0 0 7 29.4497 -98.12064 no mussels 0 0 0

9 29.44798 -98.12158 no mussels 0 0 0 FM 2538 – Zuehl Crossing 11 29.44715 -98.12095 no mussels 0 0 0 13 29.44631 -98.12479 no mussels 0 0 0

15 29.44646 -98.12554 no mussels 0 0 0 17 29.4448 -98.12896 no mussels 0 0 0 19 29.44263 -98.13298 no mussels 0 0 0

DP1 29.45148 -98.12305 no mussels 0 0 0 yellow 2 29.391564 -98.126138 0 1 1 sandshell yellow 4 29.385452 -98.121218 0 3 3 sandshell golden orb 0 1 1 6 29.38987 -98.124783 yellow 0 1 1 sandshell golden orb 1 2 3 8 29.378026 -98.122223 yellow 0 1 1 sandshell golden orb 0 1 1 Scull Crossing – FM 775 10 29.37104 -98.11535 yellow 2 2 4 sandshell louisiana 0 1 1 fatmucket 12 29.36872 -98.11538 yellow 0 6 6 sandshell yellow 14 29.365661 -98.113172 0 6 6 sandshell DP2 29.39048 -98.125684 no mussels 0 0 0 paper 0 1 1 pondshell DP3 29.38952 -98.12447 yellow 0 1 1 sandshell golden orb 1 0 1 16 29.35941 -98.10737 yellow 3 2 5 sandshell FM 775 – CR 337 yellow 18 29.3592 -98.10686 1 1 2 sandshell 20 29.35858 -98.10386 pistolgrip 0 1 1

Quantitative Qualitative Common Reach Site # Latitude Longitude No. of No. of Total Name mussels mussels

yellow 29.35858 -98.10386 1 1 2 sandshell

golden orb 2 0 2 22 29.35622 -98.1012 yellow 2 2 4 sandshell 24 29.35638 -98.10042 golden orb 0 1 1 26 29.35737 -98.08286 golden orb 0 1 1

28 29.1668 -97.99668 no mussels 0 0 0 30 29.1569 -97.994 no mussels 0 0 0 32 29.15606 -97.99112 golden orb 0 1 1

golden orb 1 0 1 FM 537 – CR225 34 29.15562 -97.99039 paper 2 0 2 pondshell yellow 36 29.15648 -97.98646 1 0 1 sandshell 38 29.13662 -97.96757 no mussels 0 0 0 40 29.12923 -97.96727 no mussels 0 0 0

21 29.01253 -97.91924 no mussels 0 0 0

golden orb 0 3 3 23 29.00923 -97.92113 pistolgrip 3 0 3 golden orb 4 1 5 25 29.00875 -97.92095 pistolgrip 0 2 2 27 29.01882 -97.91992 golden orb 1 1 2 29 29.01745 -97.92007 no mussels 0 0 0

State Hwy. 123 – 31 29.01597 -97.91924 no mussels 0 0 0 FM 2724 33 29.002 -97.91818 golden orb 1 1 2 35 28.99314 -97.92118 golden orb 0 1 1

37 28.99543 -97.91345 pistolgrip 0 1 1 39 28.99308 -97.90547 golden orb 0 5 5 41 28.99398 -97.89622 golden orb 0 1 1

DP4 29.01452 -97.92937 no mussels 0 0 0 golden orb 0 1 1 DP5 29.01484 -97.91898 yellow 0 1 1 sandshell Total - - - - 26 55 81

APPENDIX C: FOCUS ON GOLDEN ORB MUSSELS (Quadrula aurea)

The Golden Orb (Quadrula aurea) is one of 15 species of mussels listed as threatened by the Texas Parks and Wildlife Department. The United States Fish and Wildlife Service has identified 5 Central Texas Species of mussels as candidates for adding to the Lists of Endangered and Threatened Wildlife and Plants. Three of the central Texas mussel species (texas Legend Guadalupe fatmucket, texas *# Golden Orb Locations FM 2538 To Zuehl Crossing pimpleback and the Scull Crossing To FM 775 Bexar golden orb) have FM 775 To CR 337 FM 537 To CR 225 historic ranges in the *# *# SH 123 To FM 2724 San Antonio River *# *##* *# Gonzales Basin. * In response to the potential listings, the San Antonio River Authority has initiated a study to identify mussel species found in the San Antonio River Basin with specific Wilson *# interest paid to the three candidate mussel species with historic ranges in the San Antonio Basin. To date the only candidate species found by SARA *# *#*# *#Karnes staff has been the *#*#*# golden orb mussel. The o map to the right shows Atascosa0 2 4 6 8 the area’s searched and Miles Service Layer Credits: Sources: Esri, HERE, DeLorme, USGS, Intermap, increment P Corp., NRCAN, Esri Japan, METI, Esri the locations where the golden orb were found Figure 21C Locations of Golden Orb Mussels on Cibolo Creek using both qualitative and quantitative methods as described in the report.

Data collected on Cibolo Creek indicate that the most common mesohabitat types were runs. Consequently, runs were sampled more often. Of the 19 runs sampled, 10 (52.6%) had golden orbs, this was the largest percent of habitats with golden orbs. The table below shows habitat utilizations for all habitats sampled:

Number of Percent of Number of Number of Habitats with Habitats with Golden Orbs Habitat Habitat Sampled Golden Orbs Golden Orbs Found run 19 10 52.6 18 riffle 6 2 33.3 2 glide 7 1 14.3 3 pool 3 1 33.3 1 deep pool 5 1 20.0 1 backwater 6 2 33.3 6 Total 32

The most common substrates found was gravel. Consequently, gravel was sampled more often than other substrates. Of the 30 gravel substrate sampled, 12 (40.0 %) had Golden Orbs. The table below shows substrate utilizations for all substrates sampled.

Number of Number of Percent of Number of Substrate Substrates with Substrates with Golden Orbs Substrate Sampled Golden Orbs Golden Orbs Found gravel 30 12 40 21 gravel/clay 2 1 50 3 gravel/sand 4 2 50 2 sand 4 1 25 1 sand/silt 2 1 50 5 bedrock 1 0 0 0 clay 1 0 0 0 gravel/silt 2 0 0 0 Total 32

The median depth sampled was 1.4 feet, while the median depth where golden orbs were collected was 1.0 foot. The maximum depth where a golden orb was found was eight feet. The figure below shows the number of golden orbs found in the identified depth categories:

Golden Orb Depth Utilization 14

2 Number Golden OrbMussels 0 0-0.5 0.6-1.0 1.1-1.5 1.6-2.0 >2 Depth (feet)

The golden orb ranged from 26 mm to 45 mm with the median size of 35 mm. The figure below shows the number of golden orbs found in each size category:

Golden Orb - Size Classes 16 14 12 10 8 6 4 2 0 NumberGolden of OrbMussels 26-30 31-35 36-40 41-45 Size Class (mm)

As described in the report, both grid searches (quantitative) and timed searches (qualitative) were conducted along five reaches on the Lower Cibolo Creek. Golden orb mussels were found on all of the reaches except for the most upstream reach from Zuehl Crossing downstream to FM 2538 (See map 1C in Appendix C). A total of 32 golden orb mussels were found. The grid search collected a total of 11 golden orb mussels with an average population density of 0.06 golden orb per m2 and an estimate population size of 132 golden orbs. The timed searches collected a total of 21 golden orb mussels with an average catch per unit effort of 0.15 golden orb mussels per hour of search time. A total of 81 individual mussels from five native mussel species were found on Lower Cibolo Creek: yellow sandshell (38), golden orb (32), pistolgrip (7), paper pondshell (3) and Louisiana fatmucket (1). Almost 40% of the mussels found were golden orb mussels. No large clusters of mussels beds were found on the Lower Cibolo Creek but golden orbs were found at 17 out of 46 sites visited (37.0%). While native mussels including the golden orb are not common in the Lower Cibolo Creek, they are present. They do not occur in large clusters of mussels beds, but scattered sporadically throughout much of the Lower Cibolo Creek. The table below shows locations and numbers of golden orb mussels found along with population density, estimated population size and catch per unit effort:

Quantitative Qualitative Population Reach Site # Latitude Longitude Common Name No. of CPUE Size Population Density (m2) No. of mussels Total mussels (no./h) (estimates) 1 29.4531 -98.12437 no mussels 0 0 0 0 0.00 0 3 29.4505 -98.12219 no mussel 0 0 0 0 0.00 0 5 29.45027 -98.1213 no mussels 0 0 0 0 0.00 0 7 29.4497 -98.12064 no mussels 0 0 0 0 0.00 0 9 29.44798 -98.12158 no mussels 0 0 0 0 0.00 0 FM 2538 – Zuehl 11 29.44715 -98.12095 no mussels 0 0 0 0 0.00 0 Crossing 13 29.44631 -98.12479 no mussels 0 0 0 0 0.00 0 15 29.44646 -98.12554 no mussels 0 0 0 0 0.00 0 17 29.4448 -98.12896 no mussels 0 0 0 0 0.00 0 19 29.44263 -98.13298 no mussels 0 0 0 0 0.00 0 DP1 29.45148 -98.12305 no mussels 0 0 0 0 0.00 0 2 29.391564 -98.126138 no golden orb 0 0 0 0 0.00 0 4 29.385452 -98.121218 no golden orb 0 0 0 0 0.00 0 6 29.38987 -98.124783 golden orb 0 0 0 1 0.33 1 8 29.378026 -98.122223 golden orb 1 12 0.24 2 0.67 3

Scull Crossing – FM 775 10 29.37104 -98.11535 golden orb 0 0 0 1 0.33 1 12 29.36872 -98.11538 no golden orb 0 0 0 0 0.00 0

14 29.365661 -98.113172 no golden orb 0 0 0 0 0.00 0 DP2 29.39048 -98.125684 no mussels 0 0 0 0 0.00 0 DP3 29.38952 -98.12447 no golden orb 0 0 0 0 0.00 0 16 29.35941 -98.10737 golden orb 1 12 0.24 0 0.00 1 18 29.3592 -98.10686 no golden orb 0 0 0 0 0.00 0 20 29.35858 -98.10386 no golden orb 0 0 0 0 0.00 0 FM 775 – CR 337 22 29.35622 -98.1012 golden orb 2 24 0.48 0 0.00 2 24 29.35638 -98.10042 golden orb 0 0 0 1 0.33 1 26 29.35737 -98.08286 golden orb 0 0 0 1 0.33 1 FM 537 – CR225 28 29.1668 -97.99668 no mussels 0 0 0 0 0.00 0

Quantitative Qualitative Population Reach Site # Latitude Longitude Common Name No. of CPUE Size Population Density (m2) No. of mussels Total mussels (no./h) (estimates) 30 29.1569 -97.994 no mussels 0 0 0 0 0.00 0 32 29.15606 -97.99112 golden orb 0 0 0 1 0.33 1 34 29.15562 -97.99039 golden orb 1 12 0.24 0 0.00 1 36 29.15648 -97.98646 no golden orb 0 0 0 0 0.00 0 38 29.13662 -97.96757 no mussels 0 0 0 0 0.00 0 40 29.12923 -97.96727 no mussels 0 0 0 0 0.00 0 21 29.01253 -97.91924 no mussels 0 0 0 0 0.00 0 23 29.00923 -97.92113 golden orb 0 0 0 3 1.00 3 25 29.00875 -97.92095 golden orb 4 48 0.96 1 0.33 5 27 29.01882 -97.91992 golden orb 1 12 0.24 1 0.33 2 29 29.01745 -97.92007 no mussels 0 0 0 0 0.00 0 31 29.01597 -97.91924 no mussels 0 0 0 0 0.00 0 State Hwy. 123 – FM 33 29.002 -97.91818 golden orb 1 12 0.24 1 0.33 2 2724 35 28.99314 -97.92118 golden orb 0 0 0 1 0.33 1 37 28.99543 -97.91345 no golden orb 0 0 0 0 0.00 0 39 28.99308 -97.90547 golden orb 0 0 0 5 1.67 5 41 28.99398 -97.89622 golden orb 0 0 0 1 0.33 1 DP4 29.01452 -97.92937 no mussels 0 0 0 0 0.00 0 DP5 29.01484 -97.91898 golden orb 0 0 0 1 0.33 1 Total - - - - 11 132 2.64 21 7 32

Average Population ------0.06 - - - Density

Average CPUE ------0.15 -

SPECIAL THANKS:

Dr. Tim Bonner – Professor, Texas State University Dr. Charles Randklev – Texas A & M Institute of Renewable Natural Resources, College Station, TX. (Personal Communication). Bob Howells – Bio studies, Kerrville, TX. (Personal Communication). Marsha Mays – Texas Parks and Wildlife, Conservation Outreach Program – Mussel Watch Group (Identification Confirmation). Clint Robertson – Texas Parks and Wildlife, River Studies Program, San Marcos, TX. (Personal Communication). Bio - West Hookah Team - Jube Guajardo, Jeremy Hull, Brad Littrell, and Nick Porter. SARA Field Biologists – Shaun Donovan, Doug Knabe, Karen Sablan and Chris Vaughn. SARA Staff – Mick Bartlett, Karen Bishop, Melissa Bryant, Padmini Devadoss, Michelle M. Garza, Jeanette Hernandez, Ronnie Hernandez, Charles Lorea IV, Amanda Nasto, Katherine Peché, Steve Raabe, and Rebecca Reeves SARA Interns – Taylor Quiroz and Aline Trejo Others – City Office of La Vernia, TX. and Dr. Robert Frets, Seguin TX.