GEOLOGICAL SURVEY OF ALABAMA Berry H. (Nick) Tew, Jr. State Geologist

WATER QUALITY ANALYSIS AND HABITAT THREATS CONCERING CAMBARUS CRACENS ON SAND MOUNTAIN IN NORTHEAST ALABAMA

by

Rebecca A. Bearden, Anne Wynn, Pat O’Neil, and Stuart W. McGregor

with geochemical analyses by Mirza A. Beg, Rick Wagner, and Robert E. Meintzer

Prepared in cooperation with U.S. Department of the Interior U.S. Fish and Wildlife Service Alabama Ecological Services Field Office 1208-B Main Street Daphne, AL 36526

Tuscaloosa, Alabama 2017

CONTENTS Measurement Conversion Chart ...... v Executive Summary ...... vii Introduction ...... 1 Life History ...... 3 Water Quality Threats ...... 4 Acknowledgments...... 5 Study Area ...... 6 Methods ...... 11 Crayfish Surveys ...... 11 Water Quality ...... 12 Pesticides and Bacteria ...... 13 Land Use Analysis ...... 17 Results and Discussion ...... 17 Crayfish Surveys ...... 17 Water Quality ...... 18 High Flow ...... 21 Low Flow ...... 22 Pesticides ...... 23 High Flow ...... 24 Intermediate Flow ...... 24 Low Flow ...... 24 Bacteria ...... 25 High Flow ...... 25 Intermediate Flow ...... 26 Low Flow ...... 26 Land Use Analysis ...... 26 Conclusions ...... 31 Water Quality ...... 31 Land Use Analysis ...... 33 Crayfish Surveys ...... 33 Recommendations ...... 33 References Cited ...... 34 Appendix A ...... 39 Water Quality Data Collected from Sampling Sites for the 2015-2016 Cambarus cracens Survey Conducted in Scarham, Short, and Town Creek Watersheds in the Sand Mountain Area of Dekalb and Marshall Counties, Alabama………………...... 39 Appendix B ...... 48 Pesticide and Bacteria Data Collected from Sampling Sites for the 2015-2016 Cambarus cracens Survey Conducted in Scarham, Short, and Town Creek Watersheds in the Sand Mountain Area of Dekalb and Marshall Counties, Alabama ...... 48

iv CONTENTS—Continued

TABLES Table 1. Known historical locations for Cambarus cracens ...... 3 Table 2. Sampling locations for Cambarus cracens from a 2011 status survey and number of individuals collected ...... 4 Table 3. Cambarus halli life history data ...... 5 Table 4. Sampling locations for the 2015-17 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia ...... 8 Table 5. Water-quality parameters, lower limits of detection, and analytical methods...... 15 Table 6. Density estimates for Cambarus cracens from selected streams in the Scarham and Town Creek watersheds in the Sand Mountain area of DeKalb County, Alabama ...... 18 Table 7. Water-use classification and applicable water-quality criteria for the Scarham, Short and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama…………………...…...………………...... 19 Table 8. Farm statistics for DeKalb and Marshall Counties, Alabama, from 1950 to 2012 ...... 27 Table 9. Density of poultry farms in each HUC 12 subwatershed for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia ...... 30

FIGURES Figure 1. Crayfish sampling sites for historic Cambarus cracens collections conducted in the Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama ...... 2 Figure 2. Physiography of the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia ...... 6 Figure 3. Land use/land cover of the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia ...... 7 Figure 4. Crayfish sampling sites for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia…………………………………...... 11 Figure 5. Water quality, pesticide, and bacteria sampling sites for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia…………………………………………………………………………………………………………...... 14

iv CONTENTS—Continued Figure 6. Location of poultry farms relative to sampling sites for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia ...... 28

Figure 7. Number of poultry farms in each HUC 12 subwatershed for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia...... 29

iv iv iv MEASUREMENT CONVERSION CHART (from Hansen, 1991)

vi EXECUTIVE SUMMARY Scarham, Short, and Town Creek watersheds on Sand Mountain in northeast Alabama are important aquatic resources from several perspectives: water supply for an active agricultural area, biological diversity, and quality of life. These three watersheds are also home to the only known aquatic endemic species in this area, the Slenderclaw Crayfish, Cambarus cracens. Previously found at five sites in these three watersheds in the 1970s, recent sampling in 2011 found the species at only one site. The goal of this study was to determine the current range of this species and identify any water quality issues or habitat threats that may exist. Status surveys for Cambarus cracens were conducted at 71 sites from October 2015 to August August 2017. The species was found at five sites in the Scarham and Town Creek watersheds. Water quality surveys were conducted during April and October 2016 at 10 locations within the three watersheds. Pesticide and bacteria surveys were conducted at 44 sites during December 2015, May 2016, August 2016, September 2016, and October 2016. Eutrophication was found to be a significant water-quality problem in each watershed. Toxic trace metals and pesticides were also detected in each watershed. Positive detections were made for arsenic, zinc, and lead. Lead and zinc appear to be the only metals in concentration and distribution to warrant future consideration. Causes for the presence of lead are unknown but may reflect historical distribution or low-level storm runoff sources. The occurrence and concentration of atrazine may be of concern in all three watersheds, while the occurrence and concentration of 2,4-D may be of concern in the Scarham Creek watershed. Escherichia coli bacteria was shown to be elevated in the Town Creek watershed during flows in the month of August. Despite the water quality impairment the Scarham and Town Creek watersheds exhibit, Cambarus cracens continues to persist in these stream systems. The status of C. cracens as the only known aquatic endemic on Sand Mountain may be due to the crayfish’s unique life history and its ability to withstand impaired water quality conditions by burrowing. The presence of both adult and juvenile C. cracens in the upper reaches of Town Creek provides evidence that the species is reproducing successfully not only in a new location in Bengis Creek but also in a new location in Town Creek. The additional discovery of C. cracens in one new location and two previously known locations in Shoal Creek are evidence that the species appears to be surviving in that Scarham Creek tributary. Continued efforts at establishing watershed projects, promoting best management practices, and monitoring future water-quality trends will help to insure the integrity of water quality in these tributaries and can assist in improving habitat quality throughout the Scarham, Short, and Town Creek watersheds.

viii WATER QUALITY ANALYSIS AND HABITAT THREATS CONCERING CAMBARUS CRACENS ON SAND MOUNTAIN IN NORTHEAST ALABAMA Rebecca A. Bearden, Anne Wynn, Pat O’Neil, and Stuart W. McGregor

INTRODUCTION Widely known for its aquatic biodiversity, Alabama is not only a hotspot for fish and mussel species, it is also the most diverse region in the world for freshwater crayfishes (Neves, 1999; Taylor, 2002; Warren and others, 2000). This incredible biodiversity warrants the attention of biologists and natural resource managers seeking aquatic species conservation opportunities. A review by Taylor and others (2007) found that nearly half of the crayfish in the Southeast were in need of some conservation attention. This is especially relevant for the state of Alabama and its 96 described species of crayfish, as some of these are limited to a single drainage and are still substantially understudied. The Slenderclaw Crayfish, Cambarus cracens, is one species that was most recently found in only a single tributary in Alabama—Shoal Creek in the Sand Mountain area of DeKalb County. Historically known to occur at five sites in northeastern Alabama (figure 1, table 1), a recent survey suggests this crayfish is declining at a rapid rate (Kilburn and others, 2014). Because of this apparent decline, it is currently listed as endangered according to American Fisheries Society criteria (Taylor and others, 2007). Following conservation priority criteria developed by the Alabama Department of Conservation and Natural Resources (ADCNR), C. cracens is also classified as a P1 (Highest Conservation Priority) species (Shelton-Nix, 2017). This apparent trend suggests that additional research is needed to identify possible causes for population decline (Kilburn and others, 2014). Except for its original description by Bouchard and Hobbs (1976), very little is known of the Slenderclaw Crayfish. It is known only from Alabama, and its range is limited to six total sites in southeastern tributaries of Guntersville Lake (Tennessee River) in DeKalb and Marshall Counties, Alabama. The type locality of the species is Short Creek at Alabama Highway 75, 1.1 miles southwest of the junction with Alabama Highway 68 in Marshall County, Alabama (Hobbs, 1989). Bouchard and Hobbs (1976) described the habitat at the type locality as a clear, slow flowing stream with bedrock and sandy substrate, and large rocks throughout. First form males range in carapace length from 24.7 mm to 37.3 mm (Hobbs, 1989). Bouchard and Hobbs (1976) described Cambarus cracens as the sole member of the subgenus Exilicambarus. According to Owen and others (2015), C. cracens is most closely related to Cambarus halli in the subgenus Depressicambarus. The failure of Kilburn and others (2014) to find the species at any of the six historical sites reported by Bouchard and Hobbs (1976) mirrored the results of surveys conducted in 2005 by Guenter Schuster (unpublished data) and in 2009 by Taylor and Schuster (unpublished data). In addition, the type locality was intensively sampled by Dillman and six field assistants in 2007 and Cambarus cracens was not collected (C. Dillman, Virginia Institute of Marine Science, Gloucester Point, Virginia, personal communication, June 12, 2015). Even with the addition of new survey points beyond the historical locations, Kilburn and others (2014) did not find other populations

1

Figure 1. Crayfish sampling sites for historic Cambarus cracens collections conducted in the Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama.

(table 2). Cambarus cracens is now known to occur at a single site: Shoal Creek at DeKalb County Highway 372. The reasons for the decline of C. cracens are unknown since sampling locations contained suitable habitat with proper substrate and low siltation (Kilburn and others, 2014). Furthermore, Kilburn and others (2014) reported that riparian vegetation along both banks was intact at all sites sampled and no obvious signs of high nutrient loads were present. However, an abundance of poultry farms in the immediate vicinity of most historical C. cracens locations and in the south-central portion of Sand Mountain in general was noted (Kilburn and others, 2014).

2 Table 1.—Known historical locations for Cambarus cracens (Bouchard and Hobbs, 1976). Number Date Drainage State County Location Latitude Longitude Collected 10/31/1974 Tennessee AL DeKalb Bengis Creek 34.5732 -85.7511 1 04/29/1970 Tennessee AL DeKalb Scarham Creek 34.3309 -85.9779 4 03/26/1974 Tennessee AL DeKalb Scarham Creek 34.331 -85.9783 13 11/11/1973 Tennessee AL DeKalb Scarham Creek 34.331 -85.9783 21 11/12/1973 Tennessee AL DeKalb Scarham Creek 34.3053 -85.9941 23 11/12/1973 Tennessee AL Marshall Short Creek 34.2936 -86.1633 20 11/12/1973 Tennessee AL Marshall Short Creek 34.2936 -86.1633 1 11/12/1973 Tennessee AL Marshall Short Creek 34.2936 -86.1633 1 08/21/1973 Tennessee AL Marshall Shoal Creek 34.3673 -86.1213 7

Based on their findings, Kilburn and others (2014) recommended that the Slenderclaw Crayfish be considered for listing under the Endangered Species Act of 1973 (as amended). This recommendation was based on three criteria: (1) the species has experienced a significant reduction of a native range severely restricted to begin with; (2) the species is now currently known to exist at a single site; and (3) intensive field efforts have been expended in efforts to collect Cambarus cracens across its native range and in other nearby locations with suitable habitat. They also recommended that efforts be undertaken to examine water quality conditions such as nutrient loads, heavy metals, and bacterial levels at sites within the species’ range as a possible source of habitat degradation (Kilburn and others, 2014). Additionally, Schuster recommended a more thorough survey of previously sampled streams between bridge crossings (Guenter Schuster, personal communication, June 7, 2015). Though the most recent survey conducted (Kilburn and others, 2014) attempted to determine current distribution and population status of Cambarus cracens, results were not sufficient to justify listing of C. cracens as endangered (Jeff Powell, United States Fish and Wildlife Service [USFWS], Alabama Ecological Services Field Office, Daphne, Alabama, personal communication, June 6, 2015). The goals of this study are to complete the following: (1) a review of pertinent life history information for species related to C. cracens; (2) a more thorough assessment of the current status of C. cracens in areas further upstream and downstream of bridge crossings; (3) identification of possible water quality threats causing the apparent decline of C. cracens; and (4) determination of what effect, if any, land use change and habitat alteration may have had on the presence of C. cracens.

LIFE HISTORY Concerning the life history of Cambarus cracens, no detailed information has been published. Recent genetic studies have shown that C. cracens is most closely related to Cambarus halli in the subgenus Depressicambarus (Owen and others, 2015). Life history information published for C. halli analyzed age-class structure, reproductive biology, seasonal growth, and habitat use at sites on the Tallapoosa River in Georgia (Dennard and others, 2009) (table 3). In addition to two

3 Table 2.—Sampling locations for Cambarus cracens from a 2011 status survey and number of individuals collected (Kilburn and others, 2014).

Number Date Drainage State County Location Latitude Longitude Collected 10/3/2011 Tennessee AL DeKalb Town Creek 34.5706 -85.7049 0 10/3/2011 Tennessee AL DeKalb Bengis Creek 34.5734 -85.7512 0 10/3/2011 Tennessee AL DeKalb Town Creek 34.4775 -85.8089 0 10/3/2011 Tennessee AL Jackson Bryant Creek 34.6462 -85.8437 0 10/3/2011 Tennessee AL Jackson Bryant Creek 34.66 -85.8042 0 Guntersville 10/4/2011 Tennessee AL Jackson Reservoir 34.6325 -85.9723 0 10/4/2011 Tennessee AL DeKalb Black Oak Creek 34.4348 -86.0306 0 10/4/2011 Tennessee AL DeKalb Town Creek 34.3789 -85.9895 0 10/4/2011 Tennessee AL DeKalb Scarham Creek 34.3308 -85.9779 0 10/4/2011 Tennessee AL DeKalb Scarham Creek 34.3047 -85.9924 0 10/4/2011 Tennessee AL DeKalb Scarham Creek 34.295 -86.0382 0 Little Scarham 10/4/2011 Tennessee AL DeKalb Creek 34.3063 -86.0655 0 10/4/2011 Tennessee AL DeKalb Shoal Creek 34.348 -86.1256 11 10/5/2011 Tennessee AL Marshall Short Creek 34.2939 -86.1622 0 10/5/2011 Tennessee AL Marshall Short Creek 34.2134 -86.1145 0 10/5/2011 Tennessee AL DeKalb Cross Creek 34.2389 -86.0759 0 allopatric population sites, C. halli interactions with its congener Cambarus englishi were recorded at two sympatric sites. As with other stream-dwelling crayfish species, critical periods occur in the spring and fall, with pools, riffles, and runs being favored habitat types.

WATER QUALITY THREATS Reports and summaries produced by the Alabama Department of Environmental Management (ADEM) show evidence that two of the streams previously sampled for Cambarus cracens, Town Creek and Scarham Creek, were listed as impaired on the Alabama 303(d) list (ADEM, 1996a, 1998). Town Creek remains on the current list for mercury contamination due to atmospheric deposition (ADEM, 2016). The Alabama Department of Environmental Management developed a total maximum daily load (TMDL) for organic enrichment and dissolved oxygen in Town Creek (ADEM, 1996b), a TMDL for low dissolved oxygen and organic loading in Scarham Creek (ADEM, 2002a), and a TMDL for pesticides in Scarham Creek (ADEM, 2002b). A 1995 ADEM study showed that since 1993, one station at Scarham Creek lost 12 species of vital macroinvertebrates (ADEM, 1995). Additionally, a 2009 ADEM Monitoring Summary for Scarham Creek revealed that the macroinvertebrate community was in poor condition and that water quality monitoring indicated high dissolved metals, pathogens, and nutrient concentrations in

4 Table 3.—Cambarus halli life history data (Dennard and others, 2009).

Age Class Age 0—hatching began in March and April—5 mm in length Structure Age 1—graduation from 0 to 1 occurred in April/May—19 mm in length Age 2—graduation from 1 to 2 occurred in April/May—30 mm in length Carapace length frequency distribution suggested age-2 individuals were either dying in October/November or were indistinguishable from age 1 Reproductive June to September—both males and females were in a nonreproductive state Biology October—16.8 percent were Form I (reproductive) males; smallest male collected was 12 mm in length January—females were reproductively active March/April—females extruded eggs; smallest female collected was 13 mm in length May—maximum number of ovigerous females present June—100 percent of males were in nonreproductive (Form II) state Proportion of age-1 and age-2 reproductive individuals was greater at allopatric sites Number of pleopod eggs carried by females ranged from 65-217; positively correlated to female carapace length Egg diameter ranged from 2.280 to 2.476 mm; weakly positively related to female carapace length Seasonal Age 0 growth rate--0.58 mm greater at sympatric sites than allopatric sites growth Age-1 grew 2.39 mm per month at allopatric sites; age-0 and age-2 were significantly lower Age-2 growth rate--0.97 mm greater at sympatric sites than allopatric sites On average, C. halli were 2.07 mm smaller at sympatric sites than allopatric sites Habitat use Density estimates greatest during summer and lowest during spring and density Density lower at sympatric sites estimates Density of all age classes greatest in runs and pools at allopatric sites Density lower in pools and runs and greatest in riffles at sympatric sites

that stream (ADEM, 2009). Further evidence of impairment was noted in fish data provided by the Tennessee Valley Authority (TVA): “The Sand Mountain-Guntersville Reservoir Biological Monitoring 1988 Index of Biotic Integrity” technical report showed that both Town and Scarham Creeks featured a low proportion of simple lithophils (Saylor, 1989). This may indicate that habitat for species that rely on rocks and riffles could be lacking. Bouchard and Hobbs (1976) described the preferred habitat of C. cracens as a clear, slow flowing stream with bedrock and sandy substrate, and large rocks throughout. Further investigation is warranted regarding the effects of point source pollution in both Scarham and Town Creek watersheds. ACKNOWLEDGMENTS A study of this type requires the expert assistance of field and laboratory scientists of varied backgrounds. Crayfish identification was provided by Guenter Schuster, formerly of Eastern Kentucky University, and Chris Taylor, University of Illinois. Field assistance in the collection of water samples was provided by Sandi Stanley, formerly of the Geological Survey of Alabama. Assistance in obtaining collection data and GIS files relating to the surveyed watersheds was provided by Gregory Pierce of the Geological Survey of Alabama (GSA) and David Herder, GSA student intern at the University of Alabama.

5 STUDY AREA This study took place on the Sand Mountain area of northeastern Alabama in DeKalb, Etowah, Jackson, and Marshall Counties and northwestern Georgia in Dade and Walker Counties (figure 2). The Sand Mountain district is one of five districts that comprise the Cumberland Plateau physiographic region in Alabama. This district is a large plateau underlain by sandstone and shale. It ranges from 8 to 18 miles in width, and approximately 80 miles long, and gradually merges with the Warrior Basin in the southwest and Blount Mountain in the southeast. Major Tennessee River tributaries on Sand Mountain include Short Creek, South Sauty Creek, and Town Creek.

Figure 2. Physiography of the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

6 Many streams cut deep gorges through or fall precipitously over the edge of Sand Mountain. Land use is this area is primarily agricultural (figure 3). Impaired surface-water quality, a concern in the Sand Mountain area, is influenced by agricultural runoff and permitted discharges from small communities and industries. The upland nature of streams and generally poor water quality on Sand Mountain result in a less diverse fish community in this region (Mettee and others, 2002).

Figure 3.—Land use/land cover in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

7 Study sites for this project included locations upstream and downstream from those visited by Kilburn and others (2014), as well as additional sites within the Scarham Creek, Short Creek, Town Creek, Coon Creek, Bryant Creek, Long Island Creek, Big Spring Creek, Locust Fork, Big Wills Creek, and Little River watersheds in Alabama, and the Long Island Creek, Lower Nickajack Lake, Lookout Creek, and Chattanooga Creek watersheds in Georgia, chosen based on watershed characteristics (table 4, figure 4).

Table 4.—Sampling locations for the 2015-17 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

Date State County Watershed Location Latitude Longitude 10/06/2015 AL DeKalb Town Cr. Bengis Creek at County Road 116 (1974 site) 34.5732 -85.7511 10/06/2015 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 (1970 site) 34.3309 -85.9779 10/06/2015 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 (1974 site) 34.3310 -85.9783 10/06/2015 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 (1973 site) 34.3310 -85.9783 10/06/2015 AL DeKalb Scarham Cr. Scarham Creek at AL Highway 227 (1973 site) 34.3053 -85.9941 10/06/2015 AL Marshall Scarham Cr. Short Creek at AL Highway 75 (1973 site) 34.2936 -86.1633 10/06/2015 AL Marshall Scarham Cr. Shoal Creek at Tick Duckett Road (1973 site) 34.3673 -86.1213 10/06/2015 AL DeKalb Scarham Cr. Shoal Creek at Oak Grove Road (2011 site) 34.3480 -86.1256 10/06/2015 AL DeKalb Scarham Cr. Shoal Creek at Baker Road 34.3638 -86.1241 02/18/2016 AL DeKalb Scarham Cr. Scarham Creek at County Road 46 34.3047 -85.9849 02/18/2016 AL DeKalb Scarham Cr. Trib to Scarham Creek at County Road 46 34.3007 -85.9879 02/18/2016 AL DeKalb Scarham Cr. Mooney Branch at County Road 20 34.3355 -85.9858 02/18/2016 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 34.3312 -85.9786 02/18/2016 AL DeKalb Scarham Cr. Little Scarham Creek at County Road 246 34.3418 -86.0086 02/18/2016 AL DeKalb Town Cr. Reedy Creek at County Road 45 34.3694 -85.9648

02/18/2016 AL DeKalb Town Cr. Trib to Town Creek at County Road 72 34.4138 -85.9112 02/18/2016 AL DeKalb Town Cr. Trib to Town Creek at County Road 429 34.4204 -85.9520 03/23/2016 AL DeKalb Town Cr. Town Creek at County Road 140 34.6967 -85.6177 03/23/2016 AL DeKalb Town Cr. Town Creek at unnamed dirt road 34.7080 -85.6032 03/23/2016 AL DeKalb Town Cr. Town Creek at County Road 138 34.6469 -85.6509 05/26/2016 AL DeKalb Town Cr. Town Creek at County Road 140 34.6966 -85.6176 05/26/2016 AL DeKalb Town Cr. Bengis Creek at County Road 40 34.6051 -85.7223 05/26/2016 AL Marshall Short Cr. Short Creek at AL Highway 75 34.2935 -86.1633 05/26/2016 AL DeKalb Town Cr. Town Creek at County Road 40 34.6105 -85.655 05/26/2016 AL DeKalb Town Cr. Bengis Creek at County Road 112 34.5512 -85.7607 05/26/2016 AL DeKalb Town Cr. Town Creek at County Road 50 34.4269 -85.8756 05/26/2016 AL DeKalb Town Cr. Blackoak Creek at AL Highway 227 34.4178 -86.0405 05/26/2016 AL Marshall Scarham Cr. Scarham Creek at AL Highway 75 34.3077 -86.1295 05/26/2016 AL Marshall Scarham Cr. Shoal Creek at Baker Road 34.3638 -86.1243 05/26/2016 AL Marshall Scarham Cr. Shoal Creeek at Oak Grove Rd 34.3475 -86.1249 05/26/2016 AL DeKalb Scarham Cr. South Fork Scarham Creek at County Road 46 34.3046 -85.9849 05/26/2016 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 34.3308 -85.9778

8 Table 4.— Sampling locations for the 2015-17 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.—Continued

Date State County Watershed Locatio Latitude Longitude n 05/26/2016 AL DeKalb Scarham Cr. Little Scarham Creek at County Road 246 34.3419 -86.0083 05/26/2016 AL DeKalb Short Cr. Cross Creek at County Road 479 34.2388 -86.0760 05/26/2016 AL Marshall Short Cr. Short Creek at County Road 26 34.2315 -86.123 07/05/2016 AL DeKalb Scarham Cr. Scarham Creek at County Road 179 34.2985 -86.0773 07/05/2016 AL Marshall Short Cr. Trib to Short Creek at County Road 543 34.2589 -86.1232 07/05/2016 AL Marshall Short Cr. Short Creek at Rice Mill Chavers Road 34.3112 -86.2117 07/05/2016 AL Marshall Scarham Cr. Whippoorwill Creek at Pea Ridge Road 34.3257 -86.1489 07/05/2016 AL Marshall Scarham Cr. Scarham Creek at County Road 82 34.2950 -86.0963 07/05/2016 AL Marshall Scarham Cr. Scarham Creek at McVille Road 34.2985 -86.1165 07/05/2016 AL Marshall Scarham Cr. Scarham Creek at Martling Road 34.3262 -86.1615 07/05/2016 AL DeKalb Scarham Cr. Scarham Creek at AL Highway 227 34.3053 -85.9941 07/05/2016 AL DeKalb Short Cr. Short Creek at Martling Road 34.3013 -86.1809 08/16/2016 AL DeKalb Town Cr. Town Creek at County Road 140 34.6967 -85.6177

08/16/2016 AL Jackson Long Island Cr. Miller Creek at AL Highway 71 34.8345 -85.6433

08/16/2016 AL Jackson Long Island Cr. Guest Creek at AL Highway 73 34.8949 -85.6204

08/16/2016 AL Jackson Coon Cr. Coon Creek at AL Highway 171 34.7716 -85.7071

08/16/2016 AL Jackson Bryant Cr. Bryant Creek at AL Highway 71 34.6462 -85.8437

08/16/2016 AL Jackson Bryant Cr. Bryant Creek at County Road 151 34.6600 -85.8042

08/16/2016 AL DeKalb Town Cr. Caney Creek at County Road 1906 34.495 -85.8022 08/16/2016 AL DeKalb Town Cr. Caney Creek at County Road 155 34.5068 -85.8111 08/31/2016 AL Marshall Short Cr. Short Creek at County Road 26 34.2315 -86.1231 08/31/2016 AL DeKalb Short Cr. Cross Creek at County Road 479 34.2389 -86.0759 08/31/2016 AL DeKalb Scarham Cr. Scarham Creek at AL Highway 227 34.3045 -85.9925 08/31/2016 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 34.3306 -85.9783 08/31/2016 AL Marshall Short Cr. Short Creek at AL Highway 75 34.2935 -86.1633 8/7/2017 AL Marshall Scarham Cr. Scarham Creek at AL Highway 75 34.3077 -86.1295 8/7/2017 AL DeKalb Scarham Cr. Scarham Creek at AL Highway 227 (1973 site) 34.3053 -85.99413 8/7/2017 AL DeKalb Scarham Cr. Scarham Creek at County Road 20 (1973 site) 34.3310 -85.97829 8/7/2017 AL Marshall Short Cr. Short Creek at AL Highway 75 (1973 site) 34.2936 -86.16332 8/8/2017 AL Marshall Big Spring Cr. Big Spring Creek near AL Hwy. 79 34.2253 -86.4004

8/8/2017 AL Marshall Locust Fork trib to Slab Creek at Co. Hwy. 152 nr. Albertville 34.2250 -86.27155

8/8/2017 AL Marshall Locust Fork Lemon Branch at Whitesville Road 34.20559 -86.24483 8/8/2017 AL Marshall Short Cr. Drum Creek at Rose Road 34.29795 -86.22798 8/8/2017 AL Marshall Big Spring Cr. Big Spring Creek nr. AL Highway 79 34.22851 -86.39905 8/8/2017 AL DeKalb Scarham Cr. Scarham Creek at County Road 46 34.3047 -85.98491 8/9/2017 AL DeKalb Scarham Cr. Little Scarham Creek at County Road 246 34.3418 -86.00864 8/9/2017 AL DeKalb Scarham Cr. Little Scarham Creek at County Road 246 34.3419 -86.0083

9 Table 4.— Sampling locations for the 2015-17 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.—Continued

Date State County Watershed Locatio Latitude Longitude n 8/9/2017 AL Marshall Scarham Cr. Shoal Creek at Tick Duckett Road (1973 site) 34.3673 -86.12134 8/9/2017 AL DeKalb Scarham Cr. Scarham Creek at County Road 179 34.2985 -86.07725 8/9/2017 AL Marshall Scarham Cr. Scarham Creek at Martling Road 34.3262 -86.1615 8/9/2017 AL DeKalb Town Cr. Town Creek at County Road 140 34.6967 -85.61767 8/9/2017 AL Marshall Short Cr. Wolf Creek at Blessing Road 34.25181 -86.1569 8/9/2017 AL Marshall Locust Fork Slab Creek at Mt Vernon Drive 34.21066 -86.20006 8/10/2017 AL DeKalb South Sauty Cr. Piney Creek at County Highway 92 34.4847 -85.9063 8/10/2017 AL DeKalb Bryant Cr. Sandy Ford Creek at County Highway 131 34.6789 -85.7354 8/10/2017 AL DeKalb Coon Cr. Boydston Creek at County Highway 141--Willow Road 34.7298 -85.6703 8/11/2017 AL DeKalb Coon Cr. Steele Creek at County Highway 159 34.7663 -85.6481

8/11/2017 AL Jackson Long Island Cr. Guest Creek at County Highway 131 34.9142 -85.6523 8/14/2017 GA Dade Long Island Cr. Bullard Branch at Bible Camp Road 34.8460 -85.5651 8/14/2017 GA Dade Long Island Cr. Big Spring Branch at Gunsmoke Trail 34.8714 -85.5567 8/14/2017 GA Dade Lower Nickajack Lake Murphy Spring at Murphy Hollow Road 34.9590 -85.5068

8/14/2017 GA Dade Lower Nickajack Lake trib to Cole City Creek at Walls Circle 34.9479 -85.5476

8/15/2017 AL DeKalb Town Cr. Town Creek at AL 40 34.6105 -85.65496 8/15/2017 GA Walker Chattanooga Cr. Rock Creek at Rock Creek Road 34.9052 -85.40097 8/15/2017 GA Dade Lookout Cr. Gulf Creek at Holder Loop Road 34.7534 -85.4886 8/15/2017 GA Dade Lookout Cr. Allison Creek at Deer Head Cove Road 34.7713 -85.5568 8/15/2017 GA Dade Lookout Cr. Squirrel Town Creek at New England Road 34.9106 -85.4739 8/15/2017 GA Dade Lookout Cr. Crawfish Creek at Byrds Chapel Drive 34.8113 -85.5481 8/16/2017 AL DeKalb Big Wills Cr. Little Wills Creek at County Highway 51 34.2751 -85.8823 8/16/2017 AL DeKalb Short Cr. Cross Creek at County Highway 386 34.2379 -86.0973 8/16/2017 AL Etowah Big Wills Cr. Fisher Creek at Sand Valley Road 34.1106 -86.0764 8/16/2017 AL Marshall Short Cr. Short Creek at AL Highway 75 (1973 site) 34.2936 -86.16332 8/16/2017 AL DeKalb Scarham Cr. Laurel Creek at County Highway 57 34.3549 -85.9011 8/16/2017 AL DeKalb Town Cr. Bengis Creek at County Highway 116 34.5732 -85.75107 8/17/2017 AL DeKalb Little River Johnnie's Creek at County Highway 115 34.3182 -85.7174 8/17/2017 AL DeKalb Little River Indian Falls at Desoto State Park, Co. Hwy. 89 34.4991 -85.6179

8/17/2017 AL DeKalb Little River Hurricane Creek at Oakhill Road NE 34.4662 -85.6238 8/17/2017 AL DeKalb Little River Armstrong Creek at County Highway 1941 34.5188 -85.57199 8/17/2017 AL DeKalb Little River Yellow Creek at Pumpkin Center Road NE 34.4400 -85.65668 8/18/2017 AL DeKalb Scarham Cr. Shoal Creek at Oak Grove Road 34.3480 -86.1256 8/18/2017 AL Marshall Short Cr. Short Creek at AL Highway 75 (1973 site) 34.2936 -86.16332 8/18/2017 AL DeKalb Town Cr. Blackoak Creek at AL Highway 227 34.4178 -86.0405

10

Figure 4. Crayfish sampling sites for the 2015-2017 Cambarus cracens survey conducted various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

METHODS

CRAYFISH SURVEYS During October 2015, February, March, May, July, and August 2016, and August 2017, field surveys for Cambarus cracens were conducted in the following streams of northeastern Alabama and northwestern Georgia: Town Creek, Bengis Creek, Caney Creek, Reedy Creek, and Blackoak Creek in the Town Creek watershed in Alabama; Scarham Creek, Little Scarham Creek, South Fork Scarham Creek, Mooney Branch, Whippoorwill Creek, Laurel Creek, and Shoal Creek in the Scarham Creek watershed in Alabama; Short Creek, Cross Creek, Drum Creek, and Caney Creek in the Short 11 Creek watershed in Alabama; Miller Creek and Guest Creek in the Long Island Creek watershed in Alabama; Coon Creek, Boydston Creek, and Steele Creek in the Coon Creek watershed in Alabama; Bryant Creek and Sandy Ford Creek in the Bryant Creek watershed in Alabama; Big Spring Creek in the Big Spring Creek watershed in Alabama; Little Wills Creek and Fisher Creek in the Big Wills Creek watershed in Alabama; Slab Creek and Lemon Branch in the Locust Fork watershed in Alabama; Piney Creek in the South Sauty Creek watershed in Alabama; Johnnie’s Creek, Indian Falls, Hurricane Creek, Armstrong Creek and Yellow Creek in the Little River watershed in Alabama; Bullard Branch and Big Spring Branch in the Long Island Creek watershed in Georgia; Murphy Spring and Cole City Creek in Lower Nickajack Lake watershed in Georgia; Rock Creek in the Chattanooga Creek watershed in Georgia; and Gulf Creek, Crawfish Creek, Allison Creek, and Squirrel Town Creek in the Lookout Creek watershed in Georgia. Sites visited were chosen for either known historical occurrences or as potentially new occurrences based on suitable habitat. Historical site selection and detailed locality information were obtained through museum database queries at the United States National Museum of Natural History Smithsonian Institution (USNM), the Eastern Kentucky University Crustacean Collection (EKU) and the Illinois Natural History Survey Crustacean Collection (INHS). Crayfish sampling in larger streams was conducted with dip nets while visual searches were conducted at smaller stream sites. At each site, the net was set below cobble, boulders, or woody debris and held while lifting and moving rocks and kicking and shuffling crayfish into the net. Some small stream sites (< 2 m in width) required only visual searches, which involved turning over cobble, boulders, and woody debris and hand capturing crayfish or handpicking crayfish out in the open. Cambarus cracens density estimates were conducted at selected locations by choosing a random reach of the sampling site, measuring both stream length and width of that reach, and sampling to depletion. General in-stream habitat characteristics, dominant substrate type, turbidity, current type, percent cover, depth, and width were recorded for every collection site. After collection efforts were complete, crayfish were preserved in 70 percent ethanol. Specimens were transported to the Geological Survey of Alabama (GSA) then shipped to Guenter Schuster in Richmond, Kentucky, for species verification. All species were identified, counted, sexed, and cataloged into the GSA’s crayfish collection.

WATER QUALITY Water quality samples were collected during high and low flow conditions from four stream sites in the Scarham Creek watershed, two streams sites in the Short Creek watershed, and four stream sites in the Town Creek watershed (figure 5). Sites with historic locations of Cambarus cracens were sampled, as well as headwater streams and locations downstream from historic sites. Sixty-five water quality parameters, including physical characteristics, nutrients, inorganic substances, trace metals, selected pesticides, and selected biological characteristics were determined (O’Neil and Meintzer, 1995) (table 5). The following parameters were measured in situ for each sample: dissolved oxygen (DO) was measured in milligrams per liter (mg/L) using a Yellow Springs Instruments (YSI) Model Pro 20 dissolved-oxygen meter; hydrogen-ion concentration (pH), specific conductance (measured in micro Siemens per centimeter [µS/cm]), and temperature were measured with a Horiba Water Checker Model U-10. Total residual chlorine (TRC) was measured colorimetrically with a HACH Model CN-70 chlorine test kit. A 12 collected sample was inoculated with a standard reagent powder pillow, allowed to stand for three minutes for the reaction to occur, then compared against a stream blank in the standardized color-comparison wheel. An integrated grab sample of water was collected at each station, and the following raw and filtered (0.45 µm) individual samples were transported (in Nasco whirl-pak sterilized bags or polyethylene bottles) to the GSA geochemical laboratory for analysis: one 18-oz raw water bag, one 4-oz filtered-chilled bag (4°C) for anions and alkalinity, one 4-oz filtered-acidified (pH <2.0 with sulfuric acid) bag for total dissolved phosphorus and ammonia analysis, one filtered-acidified (pH <2.0 with nitric acid) sample in a white polyethylene bottle for analysis of metals, one raw sample in an amber glass bottle for total organic carbon (TOC) analysis, one raw sample in a small clear plastic bottle for mercury analysis, and one raw- acidified (pH<2.0 with sulfuric acid) for analysis of chemical oxygen demand (COD) and total Kjeldahl nitrogen (TKN). Other water-quality parameters measured during the watershed survey included biochemical oxygen demand (BOD), the pesticides alachlor, atrazine, 2,4-D, and glyphosate, total coliform bacteria, and Escherichia coli (E. coli). A 40 milliliter (mL) raw sample was collected in a commercially prepared brown-glass sample bottle for analysis of the four pesticides. The organic analysis technique of immunoassay was used to measure the four pesticide constituents. The assay reagent kits were manufactured by Ohmicron with samples analyzed according to their protocol. Two standard BOD bottles were filled using a BOD sampler and immediately chilled and kept from light during transport to the laboratory.

PESTICIDES AND BACTERIA Water samples were collected for pesticides and bacteria at 44 total sites throughout the Scarham, Short, and Town Creek watersheds at various times during high, intermediate, and low flows (figure 5). Certain sites were selected for repeated sampling based on their status as a historical or current Cambarus cracens collection site. Repeated sampling and the addition of new sites were also based on results from the previous sampling periods. Bacteria samples were collected in sterile containers and returned within a 5-hour period to the GSA Geochemical Laboratory where they were prepared, incubated, and quantified for total coliform bacteria (colony forming units per 100 milliliter of sample) (cfu/100 mL) and for E. coli bacteria (cfu/100 mL). The GSA Geochemical Laboratory used the IDEXX defined-substrate Colilert technology to assay for bacteria. This technology is capable of simultaneously detecting total coliforms and E. coli bacteria within a 24-hour period. During each bacteria sampling period, additional water-quality measurements were made including temperature (°C), specific conductance (µS/cm), pH (standard units), and turbidity (NTU).

13

Figure 5. Water quality, bacteria and pesticide sampling sites for the 2015-2017 Cambarus cracens survey conducted in the Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama.

Total coliform, fecal coliform, and E. coli are all indicators and measures of bacteria contamination in drinking water supplies and waters in contact with humans. The total coliform group is a large collection of different kinds of bacteria, the fecal coliform group is a sub-group of total coliform, and E. coli is a sub-group of fecal coliform. Generally, when a water sample is sent to a laboratory for analysis, it is tested for total coliform. If total coliform is present, the sample will also be tested for either fecal coliform or E. coli, depending on the lab testing method.

14 Table 5.—Water-quality parameters, lower limits of detection, and analytical methods

Lower limit of 1 2 Parameter Units detection Method Temperature °C -- Electrometric, field Dissolved oxygen mg/L 0.1 Electrometric, field Total residual chlorine mg/L 0.02 Colorimetric, APHA 4500-CI G pH units -- Electrometric, field Alkalinity as CaCO3 mg/L 3 Colorimetric, EPA 310.2 Specific conductance µS/cm² 1 Electrometric, field Total dissolved solids mg/L 10 Gravimetric, USGS I-1750-85 Hardness as CaCO3 mg/L 1 Calculated, USGS I-1340-85 Sulfate mg/L 0.08 Ion chromatography, EPA 300.0 Chloride mg/L 0.03 Ion chromatography, EPA 300.0 Bromide mg/L 0.05 Ion chromatography, EPA 300.0 Fluoride mg/L 0.02 Ion chromatography, USGS I-2057-85 Silica mg/L 0.06 ICP, EPA 200.7 Bicarbonate mg/L 3 Calculated, APHA 4500-CO2 D Carbonate mg/L 1 Calculated, APHA 4500-CO2 D Ammonia as N mg/L 0.02 Colorimetric, USGS I-2522-85 Total Kjeldahl nitrogen mg/L 0.07 Colorimetric, EPA 351.2 Nitrite as N mg/L 0.006 Ion chromatography, EPA 300.0 Nitrate as N mg/L 0.006 Ion chromatography, EPA 300.0 Total nitrate-nitrite as N mg/L 0.006 Ion chromatography, EPA 300.0 Total phosphorus as P mg/L 0.01 Colorimetric, EPA 365.1 Orthophosphate as PO4 mg/L 0.05 Ion chromatography, EPA 300.0 Arsenic µg/L 2 Graphite-furnace atomic absorption, EPA 200.9 Barium µg/L 1 ICP, EPA 200.7 Cadmium µg/L 0.09 Graphite-furnace atomic absorption, USGS I-1137-85 Chromium µg/L 0.8 Graphite-furnace atomic absorption, EPA 200.9 Copper µg/L 5 ICP, EPA 200.7 Iron µg/L 4 ICP, EPA 200.7 Lead µg/L 0.9 Graphite-furnace atomic absorption, EPA 200.9 Lithium µg/L 8 Graphite-furnace atomic absorption, P-E B050-5538 Manganese µg/L 0.8 ICP, EPA 200.7 Cold vapor atomic fluorescence spectrometry, EPA Mercury µg/L 0.01 245.7 Molybdenum µg/L 20 ICP, EPA 200.7 Nickel µg/L 20 ICP, EPA 200.7 Selenium µg/L 3 Graphite-furnace atomic absorption, EPA 200.9 Silver µg/L 10 ICP, EPA 200.7 (GF, EPA 200.9) Strontium µg/L 0.5 ICP, EPA 200.7 Zinc µg/L 4 ICP, EPA 200.7 Total organic carbon mg/L 0.4 Combustion, EPA 415.1 Chemical oxygen demand mg/L 30 Colorimetric, EPA 410.4 1mg/L—milligrams per liter; µ g/L—micrograms per liter; µ S/cm—microSiemens per centimeter; oC—degrees Celcius 2APHA—American Public Health Association; EPA—Environmental Protection Agency: USGS—U.S. Geological Survey; GF—graphite furnace; ICP—inductively coupled plasma spectrometry

15 Total coliform bacteria are commonly found throughout the environment in soil and vegetation and are generally harmless. If only total coliform bacteria are detected in drinking water, the source is probably environmental and fecal contamination is not likely. Fecal coliform bacteria appear in great quantities in the intestines and feces of humans and animals. The presence of fecal coliform in a water sample often indicates recent fecal contamination of the water resulting in a greater risk that pathogens are present. Most E. coli are harmless and are also found in great quantities in the intestines of humans and other warm-blooded animals. Some strains, however, may cause illness and the presence of E. coli in a drinking water sample almost always indicates recent fecal contamination and a greater risk that pathogens are present. Most E. coli outbreaks reported in the media have been related to food contamination, caused by a specific strain of E. coli known as E. coli O157:H7. When a water sample is reported as containing E. coli it does not mean that this specific strain is present and, in fact, it is probably not present. It does, however, indicate recent fecal contamination. Bacteria datasets can be highly variable, or skewed, with some samples measuring several orders of magnitude higher, or lower, than the majority of samples. Highly skewed data can unduly influence calculation of the arithmetic average by “pulling” the average value in one direction. Calculation of median values avoids this problem. Chemical analyses were conducted in accordance with the U.S. Environmental Protection Agency (USEPA 1973, 1983, 1988, 1990, 1991), Wershaw and others (1987), Fishman and Friedman (1989), and Greenberg and others (1992). Water samples were collected in accordance with the Standard Operating Procedures and Quality Assurance Manual of Alabama Department of Environmental Management (ADEM, December 1986) and the Quality Assurance-Quality Control Plan for GSA (O’Neil and Meintzer, 1995). Methods used by the GSA Geochemical Laboratory to analyze water samples are summarized in table 5. Each major instrument utilized in the laboratory was calibrated in accordance with the appropriate procedure being used. Calibration was based on high-quality analytical standards prepared in the laboratory or purchased from a reliable vendor. The calibration was checked using an appropriate check standard for the specific analytical procedure following the initial and subsequent calibrations, calibration updates, and every tenth sample processed. If the instrument was found to be grossly out of calibration following a quality control (QC) check, all samples analyzed since the last valid QC check were reanalyzed. If a proper check was not achieved after two successive attempts to calibrate, the instrument was removed from service for repair. Detection limits are established for all types of analyses made in the GSA Geochemical Laboratory. The lower limit of detection (LLD) was generally used for reporting purposes (table 5; appendices A and B). Although calculated, the instrumental detection limit (IDL) was not used for reporting purposes. The LLD represents that amount of a constituent that produces a signal of sufficient strength that 99 percent of the trials with that amount will produce a detectable signal. The LLD is determined in conjunction with determination of the IDL by multiplying the standard deviation by 3.290 to give the LLD. This method reduces the probability of Type I and Type II statistical errors to 5 percent each. The IDL represents the concentration equivalent to the analyte signal that is equal to three times the standard

16 deviation of the blank signal. The IDL is determined semiannually from the analysis of a standard solution prepared at a concentration of three to five times the estimated IDL. The IDL is the result of multiplying by three the average of the standard deviations obtained on three nonconsecutive days from seven determinations each day. Before submitting data to the project manager, all data were reviewed for correctness and completeness by using anion- cation balance, measured total dissolved solids (TDS) versus calculated TDS, and ion sum versus electrical conductivity (Greenberg and others, 1992). The scope of this investigation did not include one of the most common nonpoint source pollution problems in the watershed, namely sedimentation. Future efforts should address these types of nonpoint pollution problems with monitoring and public education efforts. These initiatives will hopefully yield positive results for water quality, biological integrity, and aesthetics for local stakeholders in the watershed.

LAND USE ANALYSIS Land use data were determined using information provided by the United States Department of Agriculture’s (USDA) National Agriculture Statistics Service (2012). Additionally, an assessment of poultry farm locations relative to sampling sites as well as poultry farm density in each subwatershed was performed using a GIS feature extraction feature. This data was placed into the geographical context of each Hydrologic Unit Code (HUC) 12 subwatershed in the study area for analysis of crayfish survey locations with the most intense poultry production. RESULTS AND DISCUSSION

CRAYFISH SURVEYS The present survey assessed 71 sites in northeastern Alabama and northwestern Georgia (figure 4, table 4). Sampling sites consisted of both historical localities and new locations. Of the 71 sampled sites, 5 were historical and 1 was the most recent Kilburn and others (2014) survey site. Cambarus cracens was found at a 1973 historical site (Shoal Creek at Tick Duckett), at a site where it was first collected in 2011 (Shoal Creek at Oak Grove Road), and at three new sites (Shoal Creek at Baker Road, Bengis Creek at County Road 40, and Town Creek at County Road 140) (fig. 4, table 6). At the 2011 site (Shoal Creek at Oak Grove Road), four specimens were found in October 2015 and two specimens in August 2017. This site had moderate flow, no turbidity, a mix of sand, cobble, and boulders, and was 0.1 m to 0.5 m deep and about 6 m wide. The C. cracens density estimate for the species at this location was 0.02/m2 (per meter squared) in 2015 and 0.01/m2 (per meter squared) in 2017.

17 Table 6.—Density estimates for Cambarus cracens from selected streams in the Scarham and Town Creek watersheds in the Sand Mountain area of DeKalb County, Alabama.

Date County Watershed Location Density 10/06/2015 DeKalb Scarham Creek Shoal Creek at Oak Grove Road 0.02/m² 10/06/2015 DeKalb Scarham Creek Shoal Creek at Baker Road 0.01/m² 05/26/2016 DeKalb Town Creek Bengis Creek at County Road 40 0.01/m² 08/16/2016 DeKalb Town Creek Town Creek at County Road 140 0.04/m² 08/08/2017 DeKalb Scarham Creek Shoal Creek at Oak Grove Road 0.01/m² 08/17/2017 DeKalb Scarham Creek Shoal Creek at Tick Duckett 0.01/m²

Another specimen was found in October 2015 upstream of the 2011 site at a new location in Shoal Creek at Baker Road. This site had low flow, no turbidity, a mix of sand, cobble, and boulders, and was 0.1 m to 0.4 m deep and about 5 m wide. The density estimate for this species at this location was 0.01/m2. In May 2016, an individual was found upstream of the 1974 historical site (Bengis Creek at County Road 116) at a new location, Bengis Creek at County Road 40. This site had moderate flow, some turbidity, a mix of sand, cobble and boulders, and was 0.1 m to 0.4 m deep and 3 m wide. The density estimate for this species at this location was 0.01/m2. In August 2016, two specimens were found at a new location, Town Creek at County Road This collection included a reproductive Form 1 male and a juvenile. This location had low flow, some turbidity, a mix of sand, cobble, and boulders, and was 0.1 m to 0.2 m deep and 3 m wide. The density estimate for this species at this location was 0.04/m2. At the 1973 historical site (Shoal Creek at Tick Duckett), one specimen was found in August 2017. This site had moderate flow, no turbidity, a mix of sand, cobble, and boulders, and was 0.3 m to 0.7 m deep and about 5 m wide. The C. cracens density estimate for the species at this location was 0.01/m2 (per meter squared).

WATER QUALITY Chemical and physical water-quality conditions in the Scarham, Short, and Town Creek watersheds can be evaluated from several perspectives including applicable water-use classifications, comparisons of parameters among tributary sites, comparisons of parameters among main channel sites, and comparisons to water-quality criteria and standards. Water- quality regimes in the system are shaped by a variety of forces including streamflow, geology, and nonpoint sources of runoff. Most pollution sources in the Scarham, Short, and Town Creek watersheds are nonpoint in origin including animal production, agriculture, and runoff from the unimproved county road systems. Results of water analyses are presented in appendix A. The Fish and Wildlife water-use classification (F&W) is applicable to all waters of the Scarham, Short, and Town Creek watersheds (ADEM, 2017). This classification has criteria limits within nine specific water quality categories (table 7). Measurements were taken at 10 sites during a high flow period in April 2016 and repeated at eight of those sites during a low flow period in October 2016 with the addition of two new sites (two of the previous high flow sites were dry).

18 Table 7.—Water-use classification and applicable water-quality criteria for the Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Criteria Fish and Wildlife (F&W) Rule 355-6-10-.09 (4) 1. Sewage, industrial wastes, or None which are not effectively treated according to rule 335- other wastes 6-10-.081 2. pH Shall not deviate more than 1 unit from ambient pH; nor be less than 6.0 or greater than 8.5 3. Temperature Maximum temperature shall not exceed 86 °F (30 °C); or maximum rise above ambient due to artificial heat shall not exceed 5 °F 4. Dissolved oxygen (DO) Daily DO shall not be less than 5.0 mg/L at all times; under extreme natural conditions DO may range between 4.0 and 5.0 mg/L 5. Toxic substances from sewage, Only such amounts as will not exhibit acute or chronic toxicity industrial wastes, or other as shown through effluent toxicity testing or by application of wastes numeric criteria given in rule 335-6-10-.072 6. Tastes, odor, and color- As #5 above producing substances attributable to sewage, industrial wastes, or other wastes 7. Bacteria Bacteria of fecal coliform group shall not exceed a geometric mean of 1,000/100 mL on a monthly average; nor exceed a maximum of 2,000/100 mL in any sample 8. Radioactivity Shall not exceed Health Department standards 9. Turbidity No turbidity, other than natural causes, that causes substantial visible contrast with natural appearance or interfere with any beneficial uses they serve; in no case shall turbidity exceed 50 NTU above background

1Rule 335-6-10-.08 (a)-(c). Waste Treatment Requirements (April 3, 1991). The following treatment requirements apply to all industrial waste dischargers, sewage treatment plants, and combined waste treatment plants. (a) As a minimum, secondary treatment or "equivalent to secondary treatment" as provided for in rules and regulations promulgated by the U.S. Environmental Protection Agency at 40 CPR Part 133 (1990), shall be applied to all waste discharges. The term "secondary treatment" is applied to biologically degradable waste and is interpreted to mean a facility which at design flow is capable of removing substantially all floating and settleable solids and to achieve a minimum removal of 85 percent of both the 5-day biochemical oxygen demand and suspended solids which, in the case of municipal wastes, is generally considered to produce an effluent quality containing a BOD5 concentration of 30 mg/L and a suspended solids concentration of 30 mg/L. Disinfection, where necessary, will also be required. Waste treatment requirements also include those established under the provisions of Sections 301, 304, 306, and 307 of the Federal Water Pollution Control Act (FWPCA). In addition the department may require secondary treatment of biologically degradable industrial wastewaters when the application of guidelines published under federal law do not produce a similar reduction in the parameters of concern. In the application of this requirement, consideration will be given to efficiencies achieved through in-process improvements. (b) In all cases an analysis of water use and flow characteristics for the receiving stream shall be provided to determine the degree of treatment required. Where indicated by the analysis, a higher degree of treatment may be required. (c) The minimum 7-day low flow that occurs once in 10 years shall be the basis for design criteria.

19 Table 7.—Water-use classification and applicable water-quality criteria for the Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama—Continued 2Rule 335-6-10-.07. Toxic pollutants applicable to State waters (May 28, 1992).

Acenaphthene 4-Chloro-3-methylphenol Dioxin (2,3,7,8- TCDD) N-Nitrosodiphenylam Acenaphthylene (PAH) 2-Chlorophenol 1,2-Diphenylhydrazine PCB-1242 (PCB) Acrolein Chromium (lll) Endosulfan (alpha) PCB-1254 (PCB) Acrylonitrile Chromium (VI) Endosulfan (beta) PCB-1221 (PCB) Aldrin Chrysene (PAH) Endrin PCB-1232 (PCB) Antimony Copper Ethylbenzene PCB-1248 (PCB) Arsenic Cyanide Fluoranthene PCB-1260 (PCB) Asbestos 4-4'-DDT Fluorene (PAH) PCB-1016 (PCB) Benzene Dibenzo (a,h) anthracene Heptachlor Pentachlorophenol (PAH) Benzidine 1,2-Dichlorobenzene Hexachlorobenzene Phenanthrene (PAH) Benzo (a) anthracene 1,3-Dichlorobenzene Hexachlorobutadiene Phenol (PAH) Benzo (a) pyrene (PAH) 1,4-Dichlorobenzene Hexachlorocyclohexane Pyrene (PAH) (alpha) 3,4-Benzofluoranthene 3,3'-Dichlorobenzidine Hexachlorocyclohexane (beta) Selenium (PAH) Benzo (ghi) perylene Dichlorobromomethane Hexachlorocyclohexane Silver (PAH) (gamma) Benzo (k) fluoranthene 1,2-Dichloroethane Hexachlorocyclopentadiene 1,1,2,2- (PAH) Tetrachloroethane Beryllium 1,1-Dichloroethylene Hexachloroethane Tetrachloroethylene Bis (2-chloroethyl) ether 2,4-Dichlorophenol lndeno (1,2,3-cd) pyrene Thallium (PAH) Bis (2-chloroisopropyl) 1,3-Dichloropropene (cis) Isophorone Toluene ether Bis (2-ethylhexyl) 1,3-Dichloropropene (trans) Lead Toxaphene phthalate Bromoform Dieldrin Mercury 1,2-Trans- dichloroethylene Cadmium 2,4-Dimethylphenol Methyl bromide 1,2,4-Trichlorobenze Carbon tetrachloride Diethyl phthalate Methyl chloride 1,1,1-Trichloroethan Chlordane Dimethyl phthalate Methylene chloride 1,1,2-Trichloroethan Chlorobenzene Di-n-butyl phthalate Nickel Trichloroethylene Chlorodibromomethane 4,6-Dinitro-2-methylphenol Nitrobenzene 2,4,6-Trichloropheno 2-Chloroethylvinyl ether 2,4 Dinitrotoluene N-NitrosodimethylamineN- Vinyl chloride Chloroform 2,4-Dinitrophenol Nitrosodi-n-propylamine Zinc

20 HIGH FLOW Dissolved oxygen ranged from 8.4 to 10.8 mg/L for all sites in April. The highest DO measurement was taken at site DEK19 (Scarham Creek at County Road 20). Dissolved oxygen saturation at this site was 125 percent based on a stream temperature of 22° C. Stream temperatures ranged from 20° C (DEK20, Bengis Creek at County Road 40) to 23° C (DEK04, Town Creek at Alabama Highway 227 and MAL09, Shoal Creek at Baker Road). Values of pH ranged from 5.0 (MAL03, Short Creek at Alabama Highway 75) to 6.7 (DEK04, Town Creek at Alabama Highway 227 and DEK17, Bengis Creek at County Road 116). Turbidity ranged from 1.5 NTU (MAL03, Scarham at Alabama Highway 75) to 4.6 NTU (DEK17, Bengis at County Road 116) in April. Alkalinity ranged from 1 mg/L at DEK19 (Scarham Creek at County Road 20) to 11 mg/L at MAL05 (Short Creek at Alabama Highway 75). Total dissolved solids ranged from 54 mg/L at MAL13 (Short Creek at County Road 26) to 125 mg/L at DEK06 (Black Oak Creek at Alabama Highway 227). Total suspended solids ranged from <4 mg/L at several sites to 142 mg/L at DEK17 (Bengis Creek at County Road 116). Biochemical oxygen demand ranged from 0.3 mg/L (DEK20, Bengis Creek at County Road 40) to 1.0 mg/L (DEK04, Town Creek at Alabama Highway 227). Ammonia concentrations throughout all three watersheds were reflective of nonpoint source pollution. All samples exceeded the background concentration level of 0.01 mg/L for uncontaminated streams, but none exceeded the toxicity limit of 0.5 mg/L. Highest concentrations were 0.216 mg/L at MAL10 (Shoal Creek at Oak Grove Road), 0.201 mg/L at MAL03 (Scarham Creek at Alabama Highway 75), and 0.195 mg/L at DEK19 (Scarham Creek at County Road 20). Nitrate was high throughout the three watersheds. Eight sites exceeded the 0.5 mg/L value established for excessive algal growth (Maidment, 1993). The highest concentration was 1.57 mg/L at MAL09 (Shoal Creek at Baker Road). Total phosphorus appeared elevated at only one site in the lower Town Creek watershed, 0.054 mg/L at DEK04 (Town Creek at Alabama Highway 227). This is the only site that exceeded the critical level necessary for excessive algae growth (0.05 mg/L). Nitrate and phosphorus at these concentrations generally indicate nonpoint sources of water quality impairment (Mays, 1996). Excessive nitrogen and phosphorus in a watershed will ultimately lead to some degree of eutrophication in the system. Arsenic was detected at eight sites during high flow and cadmium at three sites, though none of the levels were of concern for toxicity (ADEM, 2017). Chromium, copper, mercury, nickel, selenium, and silver were not detected at any of the sites. Zinc was detected at 10 sites. One site, DEK19 (Scarham Creek at County Road 20), had a value of 36.8 µg/L, which exceeded both the acute (34.97 µg/L) and chronic (35.26 µg/L) aquatic life criteria for zinc based on a water hardness of 24 mg/L (average hardness for high flow sampling period) (ADEM, 2017). Lead was detected at 9 out of 10 sites sampled. The highest concentration was measured at DEK06 (20.5 µg/L—Black Oak Creek at Alabama Highway 227) followed by DEK20 (10.0 µg/L, Bengis Creek at County Road 40), and DEK17 (5.47 µg/L, Bengis Creek at County Road 116). Although these measured concentrations do not exceed drinking water criteria, all detected measurements exceeded the chronic aquatic-life criteria for lead (0.52 µg/L) based on a water hardness of 24 mg/L. Additionally, the highest measurement (20.5 µg/L) also exceeded the acute aquatic life criteria for lead (13.26 µg/L).

21 Iron ranged from 73 to 669 µg/L, and manganese ranged from 2.5 to 166 µg/L. Barium ranged from 46.9 to 86 µg/L. Strontium ranged from 11.8 to 30.5 µg/L.

LOW FLOW Dissolved oxygen ranged from 3.2 to 7.5 mg/L for all sites in October. The lowest measurement (3.2 mg/L) was taken at MAL13 (Short Creek at County Road 26). Based on a stream temperature of 18° C, oxygen saturation was 34 percent at this site. Stream temperature ranged from 12° C (DEK22, Town Creek at County Road 140) to 20° C (MAL10, Shoal Creek at Oak Grove Road). Values of pH ranged from 5.7 (DEK22, Town Creek at County Road 140) to 7.4 (MAL09, Shoal Creek at Baker Road). Turbidity ranged from 0.6 NTU (DEK04, Town Creek at Alabama Highway 227) to 82 NTU (DEK22, Town Creek at County Road 140). At the time of sample collection, Town Creek at County Road 140 was a series of isolated pools. Alkalinity ranged from 26 mg/L at DEK04 (Town Creek at Alabama Highway 227) to 209 mg/L at DEK19 (Scarham Creek at County Road 20). Total dissolved solids ranged from 61 mg/L at DEK22 (Town Creek at County Road 140) to 265 mg/L at MAL13 (Short Creek at County Road 26). Total suspended solids ranged from <4 mg/L at several sites to 545 mg/L at DEK22 (Town Creek at County Road 140). Biochemical oxygen demand ranged from 0.8 mg/L (DEK17, Bengis Creek at County Road 116) to 4.3 mg/L (DEK08, Black Oak Creek at County Road 76). Ammonia concentrations measured across watersheds were reflective of nonpoint source pollution. Seven samples exceeded the background concentration level of 0.01 mg/L for uncontaminated streams, and three samples exceeded the toxicity limit of 0.5 mg/L (Maidment, 1993). Highest concentrations were 0.956 mg/L at DEK22 (Town Creek at County Road 140), 0.751 mg/L at DEK19 (Scarham Creek at County Road 20), and 0.563 mg/L at MAL13 (Short Creek at County Road 26). Nitrate values were high throughout the three watersheds. Five sites exceeded the 0.5 mg/L value established for excessive algal growth (Maidment, 1993). The highest concentration was 0.96 mg/L at DEK04 (Town Creek at Alabama Highway 227). Total phosphorus appeared elevated in the Town Creek watershed. The highest measured values were 0.226 mg/L at DEK08 (Black Oak Creek at County Road 76) and 0.106 mg/L at DEK22 (Town Creek at County Road 140). These two sites exceeded the critical level necessary for excessive algae growth (0.05 mg/L). Nitrates and phosphorus of these concentrations generally indicate nonpoint sources of water quality impairment (Mays, 1996). Excessive nitrogen and phosphorus in a watershed will ultimately lead to some degree of eutrophication in the system. Arsenic was detected at eight sites during low flow, though none of the levels exceeded the acute (340 µg/L) and chronic (150 µg/L) aquatic life criteria for arsenic. Zinc was detected at all 10 sites and mercury at 3 sites, though none of the levels exceeded the acute (117 µg/L) and chronic (118 µg/L) life aquatic life criteria for zinc based on a water hardness of 100 mg/L (average hardness for low flow sampling period [ADEM 2017]). Cadmium, chromium, copper, nickel, selenium, and silver were not detected at any of the sites. Lead was detected at all 10 sites. The highest concentration was measured at DEK17 (3.27 µg/L, Bengis Creek at County Road 116), followed by DEK22 (2.77µg/L, Town Creek at County Road 140). Although these measured concentrations do not exceed drinking water criteria, the two highest measurements exceeded the chronic aquatic-life criteria for lead (2.52 µg/L) based on a water hardness of 100 mg/L (ADEM, 2017). 22 Iron ranged from 44.2 to 3450 µg/L, and manganese ranged from 12.3 to 2,460 µg/L. Barium ranged from 80.5 to 159 µg/L. Strontium ranged from 34.6 to 169 µg/L.

PESTICIDES The occurrence of herbicides and pesticides in surface waters is universal; however, concentration is related to season of application and hydrologic conditions such as drought and rain events during application. This study analyzed water samples for concentrations of the following four agricultural-related pesticides: alachlor, atrazine, 2,4-D, and glyphosate. Alachlor is an herbicide used for weed control on corn, soybeans, sorghum, peanuts, and beans. Alachlor is slightly to moderately toxic on an acute basis to freshwater fish (LC50 [the concentration of a material that will kill 50% of the test subjects when administered as a single exposure] 1-33 ppm), and highly to moderately toxic to freshwater fish on a chronic basis (No Observable Effect Concentration [NOEC] >=0.1 ppm), (Lowest Observable Effect Concentration [LOEC]>=0.2 ppm) (USEPA, 1998). Atrazine is a commercial restricted-use herbicide used to control weeds in turf, commercial vegetables, pasture forage crops, and corn. It has a moderate surface loss and leaching potential and is generally only slightly toxic to aquatic life. Aquatic fauna are indirectly affected at atrazine concentrations of 20 µg/L and higher, through both a reduction in food supply of herbivores and loss of macrophyte habitat. Direct adverse effects to aquatic invertebrates and fishes were measured at 94 µg/L and higher (Eisler, 1989). Proposed criteria for aquatic life protection are <5 µg/L of atrazine for sensitive species of aquatic flora and <11 µg/L for most species of aquatic plants and animals (Eisler, 1989). The compound 2,4-D is primarily an herbicide and secondarily a plant growth regulator. Formulations include esters, acids, and several salts, which vary in their chemical properties, environmental behavior, and toxicity. Toxicity to fish and aquatic invertebrates varies widely depending on chemical form, with esters being the most toxic (World Health Organization, 1989; Tomlin, 2006). Acid and amine salt LC50 values range from greater than 80 to 2,244 mg acid equivalents per liter (mg ae/L) whereas the esters range from less than 1.0 to 14.5 mg ae/L (USEPA, 2005). Glyphosate is an herbicide used to kill broadleaf plants and grasses. The sodium salt form of glyphosate is used to regulate plant growth and ripen fruit. Glyphosate is used in agriculture and forestry, on residential lawns, and for weeds in industrial areas (USEPA, 2005). Some products containing glyphosate are used to control aquatic plants. Technical grade glyphosate ranges from slightly toxic to nontoxic to freshwater fish, with a 48-hour LC50 value greater than 24 mg/L to 140 mg/L, while formulated glyphosate products range from moderately toxic to nontoxic to freshwater fish, with 96-hour LC50 values ranging from 1.3 mg/L to greater than 1,000 mg/L (USEPA, 2005). Technical grade glyphosate is slightly toxic to practically nontoxic to freshwater invertebrates, with a 48-hour LC50 ranging from 55 ppm to 780 ppm (Folmar and others, 1979). The 48-hour LC50 for daphnids was 3.0 mg/L and the LC50 for midge larvae was 16 mg/L when exposed to the herbicide (Folmar and others, 1979). Water samples were collected for pesticides at 44 total sites throughout the Scarham, Short, and Town Creek watersheds at various times during high and low flows (figure 3). Certain sites were selected for repeated sampling based on their status as a historical or current Cambarus 23 cracens collection site. Repeated sampling and the addition of new sites were also based on results from the previous sampling periods. Thirty sites were sampled for pesticides during high flows in December 2015. Ten of those sites were resampled in May 2016 and 10 new sites were also sampled. In August 2016, during intermediate flows, 6 of the sites were chosen to be repeated from previous collections in December and May, 11 were re-sampled from sites in December, 5 sites were re-sampled from May, and 1 site was added for a total of 23 sites sampled. Subsequent sampling occurred during periods of low flow for 23 sites in September 2016. Six sites were resampled from the previous three surveys, 11 from December and August sampling sites, 5 from May and August sampling sites, and 1 was repeated from the August sampling site. The final round of sampling occurred during extreme low flows in October 2016. Ten total sites were sampled. Four were previously sampled during each of the four collection periods. One was sampled previously in December, August, and September, one was sampled previously in December and May, one was a repeat from December, one was a repeat from May, and one new collection site was added.

HIGH FLOW In December, alachlor values were less than the lower limit of detection at all of the sites sampled. Atrazine was detected at 83 percent of the sites sampled (appendix B). The highest concentration (0.193 µg/L) was reported from MAL09 (Shoal Creek at Baker Road). Glyphosate and 2,4-D values were less than the lower limit of detection at all of the sites sampled (appendix B). In May, alachlor values were less than the lower limit of detection at all of the sites sampled. Atrazine was less than the lower limit of detection at 30 percent of the sites sampled. The highest concentrations (0.43 µg/L and 0.34 µg/L) were reported from MAL09 (Shoal Creek at Baker Road) and MAL10 (Shoal Creek at Oak Grove Road), respectively. The herbicide 2,4-D was detected at 5 percent of the sites sampled. The highest concentration (1.6 µg/L) was reported from DEK24 (Town Creek at County Road 50). Glyphosate was detected at 10 percent of the sites sampled. The highest concentration (0.218 µg/L) was from DEK24 (Town Creek at County Road 50) as well.

INTERMEDIATE FLOW In August, alachlor values were less than the lower limit of detection at all of the sites sampled (appendix B). Atrazine was detected at 52 percent of the sites sampled. The highest concentrations (0.289 and 0.279 µg/L) were from MAL08 (Scarham Creek at Martling Road) and MAL03 (Scarham Creek at Alabama Highway 75), respectively. The herbicide 2,4-D was detected at 13 percent of the sites sampled. The highest concentration (3.8 µg/L) was from DEK14 (Bengis Creek at County Road 27). Glyphosate was detected at 30 percent of the sites sampled. The highest concentration (0.316 µg/L) was from MAL12 (Shoal Creek at Old Bethel Road).

LOW FLOW In September, alachlor values were less than the lower limit of detection for all of the sites sampled (appendix B). Atrazine was detected at 43 percent of the sites sampled. The highest concentrations (0.239 and 0.230 µg/L) were from MAL06 (Turkey Creek at Cochran Road) and MAL08 (Scarham Creek at Martling Road), respectively. The herbicide 2,4-D was detected at nine percent of the sites sampled. The highest concentration (17.4 µg/L) was from DEK19 (Scarham Creek at County Road 20). Glyphosate was detected at 52 percent of the sites sampled. The highest concentration (0.508 µg/L) was from MAL06 (Turkey Creek at Cochran Road). 24 In October, alachlor values were less than the lower limit of detection for all of the sites sampled (appendix B). Atrazine was detected at 60 percent of the sites sampled. The highest concentration (0.123 µg/L) was from MAL10 (Shoal Creek at Oak Grove Road). Glyphosate and 2,4-D were less than the lower limit of detection for all of the sites sampled.

BACTERIA Water-quality samples were collected at 44 total sites throughout the Scarham, Short, and Town Creek watersheds at various times during high, intermediate and low flows and analyzed for bacteria concentration (figure 3). Certain sites were selected for repeated sampling based on their status as a historical or current Cambarus cracens collection site. Repeated sampling and the addition of new sites were also based on results from previous sampling periods. Thirty sites were sampled during high flows in December 2015. Ten of those sites were resampled in May 2016 and 10 new sites were also sampled. In August 2016, during intermediate flows, six of the sites were chosen to be repeated from the previous two collections in December and May, 11 were re-sampled from sites in December, five were re-sampled from May, and one new site was added for a total of 23 sites sampled. Subsequent sampling occurred during periods of low flow for 23 sites in September 2016. Six sites were resampled from the previous three surveys, 11 from December and August sampling sites, 5 from May and August sampling sites, and 1 was repeated from the August sampling. The final round of sampling occurred during extreme low flows in October 2016. Ten total sites were sampled. Four were previously sampled during each collection period, one was sampled previously in December, August and September, one was sampled previously in December and May, one was a repeat from December, one was a repeat from May, and one was a new collection site. Stations were established in the larger tributaries and in tributaries with suspected polluted runoff. The sampling regime was designed to collect water samples during the two extremes of the hydrologic cycle: low flow and high flow. Samples collected during low flow periods represent source water originating from shallow groundwater aquifers. If bacteria from poorly maintained septic tanks, damaged sanitary sewer systems, or infiltration of animal wastes were in contact with shallow groundwater, then sampling during low flow periods should detect these problems. Samples collected during high flow periods represent source water from overland runoff of rainfall. Bacteria contamination originating from livestock or poultry operations, or from poorly managed treatment facilities, would be detected during high flow periods. The Alabama Department of Environmental Management has established that in non-coastal waters, bacteria of the E. coli group shall not exceed a geometric mean of 548 colonies/100 mL, nor exceed a maximum of 2,507 colonies/100 mL in any sample (ADEM, 2017).

HIGH FLOW In December, the concentration of E. coli bacteria ranged from 44 to 2,420 cfu (median 223). The highest reading (2,420 cfu) was from DEK03 (Short Creek at County Road 142). In May, the concentration of E. coli bacteria ranged from 21 to 365 cfu (median 65), with the highest reading from DEK27 (Little Scarham at County Road 246). None of these sites exceeded the recommended single sample maximum of 2,507 cfu. During high flows, median E. coli counts were less than 200 cfu in the Town Creek watershed and less than 300 cfu in the Scarham and Short Creek watersheds (appendix B).

25 INTERMEDIATE FLOW In August, the concentration of E. coli bacteria ranged from 2 to 5,560 cfu (median 214), with the highest readings (5,560 and 4,710 cfu) from DEK18 (Bengis Creek at County Road 678) and DEK20 (Bengis Creek at County Road 40), respectively. Three sites did exceed the single sample maximum of 2,507 cfu: DEK18 (Bengis Creek at County Road 678, 5,560 cfu), DEK20 (Bengis Creek at County Road 40, 4,710 cfu), and DEK14 (Bengis Creek at County Road 27, 3,500 cfu). During intermediate stream flows within the Town Creek watershed, including small direct tributaries, the upstream sections had higher median bacteria concentrations compared to the middle and downstream sections (appendix B). One upstream tributary in particular, Bengis Creek, had the highest bacteria concentrations (3,500 to 5,560 cfu). Only one tributary to Scarham Creek (South Fork Scarham Creek) exhibited an elevated bacteria concentration (2,420 cfu) when compared to the other tributaries. Scarham Creek at Alabama Highway 75 also showed an elevated bacteria level (2,420 cfu) when compared to upstream and downstream locations. The remainder of the Scarham Creek sites and all the Short Creek sites had a median bacteria concentration less than 200 cfu. The August sampling occurred one day after the area received just over 0.6 inch of rain. The storm event may have caused runoff over agricultural areas and created elevated bacteria levels in certain parts of the Town Creek watershed.

LOW FLOW In September, the concentration of E. coli bacteria ranged from 3 to 3,840 cfu (median 23), with the highest reading from DEK26 (South Fork Scarham Creek at County Road 46). This site did exceed the single sample maximum of 2,507 cfu. In October, the concentration of E. coli bacteria ranged from 1 to 292 cfu (median 17), with the highest reading from DEK17 (Bengis Creek at County Road 116). The October samples did not exceed the recommended single sample maximum of 2,507 cfu. During low flows, the median bacteria concentrations were similar throughout each watershed, with median E. coli counts less than 200 cfu in all three watersheds (appendix B). LAND USE ANALYSIS Data from the National Agricultural Statistics Service (USDA, 2012) was used to determine change in land use over time in DeKalb and Marshall Counties (table 8). In 1969, prior to the discovery of Cambarus cracens, land use in DeKalb and Marshall Counties was primarily agricultural with 60 and 56 percent of the total acreage in farms, respectively. These uses included activities such as pastureland, poultry production, and row crop production. In 2012, land use remained primarily agricultural, with 46 and 45 percent of the total acreage in farms, respectively. The GIS extraction feature was used to determine poultry farm proximity to sampling sites for the 2015-2017 survey (figure 6). Additionally, the number of farms per HUC 12 subwatershed was defined, and the density of farms per square kilometer was calculated (figure 7, table 9). Seven watersheds in the study area contained 18 or more farms, and four of those watersheds (Bengis Creek, Snake Creek-Town Creek, Upper Scarham Creek, and Whippoorwill Creek-Shoal Creek) are watersheds in which historic or current Cambarus cracens collections have been made. Eight watersheds in the study area had poultry farm densities greater than or equal to 0.249

26 farms per square kilometer. Of those eight subwatersheds, three (Bengis Creek, Upper Scarham Creek, and Whippoorwill Creek-Shoal Creek) are watersheds in which historic or current Cambarus cracens collections have been made.

Table 8.—Farm statistics for DeKalb and Marshall Counties, Alabama, from 1950 to 2012 (USDA, 2012)

DeKalb County Number of Acres in Avg. size of Land area in Reporting Year farms farms farm (acres) farms (%) farms 1950 7,055 395,556 56.1 79.5 NA 1954 6,136 396,548 64.6 79.6 165 1959 4,435 334,047 75.3 67.1 NA 1964 3,399 285,923 84.1 57.4 NA 1969 3,483 299,316 86 60.1 578 1974 2,694 235,685 87 47.3 351 1978 2,378 233,003 98 46.8 411 1982 2,228 221,502 99 44.5 339 1987 2,047 213,440 104 42.9 296 1992 1,894 210,733 111 42.3 270 1997 2,080 223,685 108 44.9 278 2002 2,177 237,336 109 47.7 271 2007 2,426 235,246 97 47.2 277 2012 2,035 229,294 113 46 251 Marshall County Acres in Avg. size of Land area in Reporting Year No. farms farms farm (acres) farms (%) farms 1950 6,053 305,211 50.4 83.6 NA 1954 4,858 297,271 61.2 81.3 212 1959 3,290 219,018 66.6 59.9 NA 1964 2,757 188,546 68.4 51.6 NA 1969 2,692 205,105 76 56.1 374 1974 2,015 160,500 80 43.9 220 1978 1,744 163,063 93 44.6 255 1982 1,664 151,585 91 41.8 219 1987 1,582 136,599 86 37.6 195 1992 1,364 142,873 105 39.4 174 1997 1,583 146,129 92 40.3 178 2002 1,686 160,590 95 44.2 178 2007 1,731 154,548 89 42.6 164 2012 1,505 162,980 108 44.9 153

27

Figure 6. Location of poultry farms nearest to sampling sites for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

28

Figure 7. Number of poultry farms in each HUC 12 subwatershed for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia.

29 Table 9.— Density of poultry farms in each HUC 12 subwatershed for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia

HUC 12 subwatershed Watershed Watershed Number Farms per Farms per area (acres) area (km2) of farms 100 acres km2 Bengis Creek 16,256 66 25 0.154 0.380 Big Spring Creek 30,363 123 9 0.030 0.073 Black Oak Creek 17,491 71 23 0.131 0.325 Boydston Creek-Burkhalter Creek 16,897 68 17 0.101 0.249 Bryant Creek 32,187 130 14 0.043 0.107 Coon Creek 27,750 112 2 0.007 0.018 Crawfish Creek-Lookout Creek 22,945 93 3 0.013 0.032 Cross Creek 21,259 86 22 0.103 0.256 Dry Creek-Lookout Creek 27,104 110 1 0.004 0.009 East Fork West Fork Little River-West Fork 15,752 64 0 0.000 0.000 FisherLittle RiverCreek 19,180 78 9 0.047 0.116 Flat Rock Creek 12,598 51 3 0.024 0.059 Gilbert Branch-Laurel Creek 12,632 51 0 0.000 0.000 Guest Creek-Long Island Creek 23,700 96 12 0.051 0.125 Gulf Creek-Lookout Creek 18,250 74 4 0.022 0.054 Headwaters Big Wills Creek 35,811 145 2 0.006 0.014 Hicks Creek-Bear Creek 9,949 40 1 0.010 0.025 Hog Creek 13,568 55 6 0.044 0.109 Johnnies Creek 13,195 53 4 0.030 0.075 Kirby Creek 24,164 98 5 0.021 0.051 Little Sand Valley Creek 22,722 92 16 0.070 0.174 Long Branch-Rock Creek 15,960 65 0 0.000 0.000 Long Creek-Miller Creek 31,141 126 16 0.051 0.127 Lookout Creek 26,451 107 0 0.000 0.000 Lower Nickajack Lake-Tennessee River 55,063 223 0 0.000 0.000 Lower Scarham Creek 7,777 31 8 0.103 0.254 Lower Short Creek 11,739 48 16 0.136 0.337 Lower South Sauty Creek 29,963 121 7 0.023 0.058 Middle Fork Little River 13,573 55 0 0.000 0.000 Middle Short Creek 28,365 115 8 0.028 0.070 Minky Creek-Town Creek 27,375 111 16 0.058 0.144 Reedy Creek-Town Creek 46,048 186 35 0.076 0.188 Sitton Gulch Creek-Lookout Creek 24,780 100 1 0.004 0.010 Snake Creek-Town Creek 30,677 124 18 0.059 0.145 Straight Creek 10,538 43 5 0.047 0.117 Straight Creek-West Fork Little River 16,016 65 0 0.000 0.000 Upper Big Wills Creek 53,872 218 11 0.020 0.050 Upper Little River 13,419 54 0 0.000 0.000 Upper Scarham Creek 31,239 126 32 0.102 0.253 Upper Short Creek 23,340 94 14 0.060 0.148

30 Table 9.— Density of poultry farms in each HUC 12 subwatershed for the 2015-2017 Cambarus cracens survey conducted in various watersheds in the Sand Mountain area of DeKalb, Etowah, Jackson, and Marshall Counties, Alabama, and Dade and Walker Counties, Georgia—Continued.

HUC 12 subwatershed Watershed Watershed Number Farms per Farms per area (acres) area (km2) of farms 100 acres km2 Upper Slab Creek 27,992 113 15 0.054 0.132 Upper South Sauty Creek 30,292 123 14 0.046 0.114 Whippoorwill Creek-Shoal Creek 19,572 79 21 0.107 0.265 Yellow Creek 9,565 39 1 0.010 0.026

CONCLUSIONS

WATER QUALITY The results of monitoring in the Scarham, Short, and Town Creek watersheds point to a few significant water-quality issues. The first issue is low DO levels during low flow in late summer and fall, possibly caused by eutrophication effects, elevated temperatures, and reduced stream flow. Half of the sites sampled had DO readings of 5.0 mg/L or below: MAL13 (3.2 mg/L, Short Creek at County Road 26); DEK19 (3.5 mg/L, Scarham Creek at County Road 20); MAL10 (4.0 mg/L, Shoal Creek at Oak Grove); DEK08 (Black Oak Creek at County Road 76), and MAL09 (5.0, Shoal Creek at Baker Road). Thirty percent of the sites with low DO issues are in the Scarham Creek watershed (appendix A). The second issue is potential eutrophication in late summer and fall (related to the low DO issue), due to elevated nutrient concentrations, elevated temperatures, and reduced stream flow. Ammonia concentrations recorded within all three watersheds were reflective of nonpoint source pollution. This may cause issues as elevated ammonia levels may adversely impact aquatic life. While concentrations of ammonia as N in uncontaminated streams may be as low as 0.01 mg/L, concentrations in contaminated streams may range from 0.5 to 3.0 mg/L (Maidment, 1993). Laboratory experiments demonstrated that exposure to concentrations as low as 0.002 mg/L for longer periods of time (up to six weeks) may be harmful to aquatic organisms causing hyperplasia of gill linings in salmon fingerlings, which may lead to bacterial gill disease (USEPA, 2013). Even short exposures to concentrations as low as 0.1 mg/L can lead to skin, eye and gill damage (USEPA, 2013). The highest concentrations in this study were 0.956 mg/L at DEK22 (Town Creek at County Road 140), 0.751 mg/L at DEK19 (Scarham Creek at County Road 20), and 0.563 mg/L at MAL13 (Short Creek at County Road 26). Nitrate values were relatively normal within the three watersheds. The highest concentration was 0.96 mg/L at DEK04 (Town Creek at Alabama Highway 227). Total phosphorus was more elevated in the lower Town Creek watershed. The highest measured value was 0.054 mg/L at DEK04 (Town Creek at Alabama Highway 227). The detection of toxic trace metals and pesticides is noteworthy, but their relative contribution to water-quality degradation in the watershed is likely much less than those pollutants causing eutrophication. Positive detections were made for arsenic, mercury, lead, and

31 zinc. Of the four, lead and zinc were detected in concentrations exceeding the aquatic life criteria. Lead was detected in 90 percent of the samples during high flow. The highest concentration was measured at DEK06 (20.5 µg/L, Black Oak Creek at Alabama Highway followed by DEK20 (10.0 µg/L, Bengis Creek at County Road 40), and DEK17 (5.47 µg/L, Bengis Creek at County Road 116). Although these measured concentrations do not exceed drinking water criteria, all detected measurements exceed the chronic aquatic-life criteria for lead based on a water hardness of 24 mg/L (average hardness for the high flow sampling period). Additionally, the highest measurement (20.5 µg/L) exceeded the acute aquatic life criteria for lead. Lead was detected at all 10 sites (100 percent) during low flow, with the highest concentration measured at DEK17 (3.27 µg/L, Bengis Creek at County Road 116), followed by DEK22 (2.77 µg/L, Town Creek at County Road 140). Although these measured concentrations do not exceed drinking water criteria, the two highest measurements exceed the chronic aquatic-life criteria for lead based on a water hardness of 100 mg/L (average hardness for low flow sampling period). The occurrence of lead does not appear related to any identifiable source and may reflect historical distribution due to its worldwide use in gasoline or a low-level storm runoff source. The occurrence and concentration of lead should be evaluated more thoroughly in future water- quality surveys. The rate of occurrence and concentrations of glyphosate appeared comparable to similar watersheds. The occurrence and concentration of atrazine may be of concern for the Scarham Creek watershed as concentrations were highest at MAL08 (Scarham Creek at Martling Road), with 0.289 µg/L. The occurrence and concentration of 2,4-D may be of concern for one site, DEK 19 (Scarham Creek at County Road 20), with the highest concentration measured during this study (17.4 µg/L). The biologically critical headwater tributaries of Scarham and Town Creek watersheds were evaluated during both low and high stream flow periods. One tributary of Town Creek, Bengis Creek, was found to be especially impaired due to nutrients and bacteria, as did tributaries of Scarham Creek and other sites within the Scarham Creek watershed. Toxic trace metals and selected pesticides were evaluated on a watershed basis and, although found mostly in low concentrations, may warrant selective monitoring in the future, particularly for lead and atrazine. Reports and summaries produced by ADEM have listed Town Creek and Scarham Creek as impaired on the Alabama 303(d) list (ADEM, 1996a, 1998), a TMDL for organic enrichment and dissolved oxygen in Town Creek (ADEM, 1996b), a TMDL for low dissolved oxygen and organic loading in Scarham Creek (ADEM, 2002a), and a TMDL for pesticides in Scarham Creek (ADEM, 2002b) have been developed. A 1995 ADEM study showed that since 1993, one station at Scarham Creek lost 12 species of vital macroinvertebrates (ADEM, 1995). Additionally, a 2009 ADEM monitoring summary for Scarham Creek revealed that the macroinvertebrate community was in poor condition and that water quality monitoring indicated high dissolved metals, pathogens, and nutrient concentrations in that stream (ADEM, 2009). Further evidence of impairment is noted in fish data provided by TVA. The Sand Mountain- Guntersville Reservoir Biological Monitoring 1988 Index of Biotic Integrity technical report showed that both Town and Scarham Creeks featured a low proportion of simple lithophils (Saylor, 1989). This may indicate that habitat for species that rely on rocks and riffles could be lacking. Bouchard and Hobbs (1976) described the preferred habitat of Cambarus cracens as a clear, slow flowing stream with bedrock and sandy substrate, and large rocks throughout.

32 LAND USE ANALYSIS Land use in DeKalb and Marshall Counties has remained primarily agricultural since Cambarus cracens was discovered in 1970, and no evidence of significant habitat alteration has been found. Intense poultry production was found to be present throughout much of the study area, with areas of higher poultry farm density located in subwatersheds where C. cracens was historically and most recently collected.

CRAYFISH SURVEYS The failure of researchers to find the species in the main channel of Scarham Creek or in the Short Creek watershed may be due to several factors: 1. Sampling bias. Researchers were unable to access potential habitat beneath larger boulders. 2. Microlandscape habitat alteration. Factors limiting distribution maybe on a more local scale than what has been analyzed in this study. 3. Species carrying capacity. The current distribution differs little from the distribution when the species was first discovered in the1970s. 4. Genetic limitations. The low number of individuals for this narrow endemic could be due to low genetic diversity. RECOMMENDATIONS The Scarham and Town Creek watersheds still provide a refuge for Cambarus cracens and quality of water in streams draining this area should be maintained at a reasonable level and improved when possible to ensure the continued existence of this rare species. Chemical water quality is one critical aspect of the integrated aquatic ecosystem that supports and maintains populations in the area. Results reported in this study documented impaired water quality conditions in some areas and measured those parameters suspected of causing impairment. To better maintain and improve water-quality conditions in the Scarham, Short, and Town Creek watersheds, the following recommendations are offered: 1. Measure the extent and identify sources of ammonia in the watershed. The elevated levels are cause for concern and should be investigated within the goals of a watershed management and protection program to ameliorate the sources. 2. Establish long-term water quality monitoring sites within the watersheds. Long- term trend sites can assist in assessing the effectiveness of watershed management activities and in maintaining water quality in the critical upper tributary reaches where Cambarus cracens is currently found. 3. Increase collaboration with the currently established USDA Natural Resources Conservation Service (NRCS) 2017 National Water Quality Initiative (USDA, 2017). The USDA NRCS has listed the Scarham Creek watershed as part of this initiative. Through this program, the NRCS works with farmers to reduce nitrogen, phosphorous, sediment, and pathogen contributions from agricultural land. This program requires instream water quality monitoring by state agencies (ADEM) as well as the USEPA. 4. Work with U.S. Fish and Wildlife Service Partners for Fish and Wildlife Program to establish landowner contacts and seek landowner input regarding habitat management practices that will foster improved water quality.

33 REFERENCES CITED Alabama Department of Environmental Management, 2017, Water Division, Water Quality Program, Vol. 1, Division 335-6: Montgomery, Alabama Department of Environmental Management. Alabama Department of Environmental Management, 1986, Standard operating procedures and quality assurance manual: Montgomery, Alabama, Alabama Department of Environmental Management. Alabama Department of Environmental Management, 1995, Sand Mountain/Lake Guntersville watershed project, macroinvertebrate bioassessment, May 31-June 1, 1994: Montgomery, Alabama Department of Environmental Management, Field Operations Division, Ecological Studies Section, 41 p. Alabama Department of Environmental Management, 1996a, Final 303 (d) list for Alabama streams: Montgomery, Alabama Department of Environmental Management, Water Division, Water Quality Branch, 4 p. Alabama Department of Environmental Management, 1996b, TMDL, Town Creek Tennessee River basin, organic enrichment/dissolved oxygen, ammonia: Montgomery, Alabama Department of Environmental Management Water Division, Water Quality Branch, 14 p. Alabama Department of Environmental Management, 1998, Final 303 (d) list for Alabama streams: Montgomery, Alabama Department of Environmental Management, Water Division, Water Quality Branch, 11 p. Alabama Department of Environmental Management, 2002a, Final TMDL development for Scarham Creek, AL/06030001-270_01, Low dissolved oxygen/ organic loading, ammonia as nitrogen: Montgomery, Alabama Department of Environmental Management Water Division, Water Quality Branch, 44 p. Alabama Department of Environmental Management, 2002b, Final Big Nance Creek and Scarham Creek, TMDL development for Pesticides (insecticides and herbicides): Montgomery, Alabama Department of Environmental Management, Water Division, Water Quality Branch, 29 p. Alabama Department of Environmental Management, 2009, Rivers and streams monitoring program, 2009 monitoring summary, Scarham Creek at Marshall County Road 371: Montgomery, Alabama Department of Environmental Management Water Division, Water Quality Branch, 2 p. Alabama Department of Environmental Management, 2016, Final 303 (d) list for Alabama streams: Montgomery, Alabama Department of Environmental Management, Water Division, Water Quality Branch, 13 p. Bouchard, R. W., and Hobbs, H. H., Jr., 1976, A new subgenus and two new species of crayfishes of the genus Cambarus (Decapoda: Cambaridae) from the southeastern United States: Smithsonian Contributions to Zoology, v. 224, p. 1-15. Dennard, S., Peterson, J. T., and Hawthorne, E. S., 2009, Life history and ecology of Cambarus halli (Hobbs): Southeastern Naturalist, v. 8, no. 3, p. 479-494. Eisler, R., 1989, Atrazine hazards to fish, wildlife, and invertebrates: a synoptic review: U.S. Fish and Wildlife Service Biological Report v. 85, no 1.18, 53 p.

34 Fishman, M. J., and Friedman, L. C., eds., 1989, in Methods for determination of inorganic substances in water and fluvial sediments: U.S. Geological Survey, Techniques of Water Resources Investigations, Book 5, Chapter A1, 545 p. Folmar, L. C, Sanders, H. O., Julin, A. M., 1979, Toxicity of the herbicide glyphosate and several of its formulations to fish and aquatic invertebrates: Archives of Environmental Contamination and Toxicology, v. 8, p. 269-278. Greenberg, A. E., Clesceri, L. S., and Eaton, A. D., eds., 1992, Standard methods for the examination of water and wastewater: Washington, D.C., American Public Health Association. Hansen, W. R., editor, 1991, Suggestions to Authors of the Reports of the United States Geological Survey, 7th ed.: Washington, DC, U.S. Geological Survey Suggestions to Authors, p. 123.Hobbs, H. H., Jr., 1989, An illustrated checklist of the American crayfishes (Decapoda: Astacidae, Cambaridae, and Parastacidae): Smithsonian Contributions to Zoology, v. 480, p. 1-236. Kilburn, S. L., Taylor, C. A., and Schuster, G. A., 2014, Conservation assessment and habitat notes for three rare Alabama crayfishes: Cambarus cracens, Cambarus scotti, and Cambarus unestami: Southeastern Naturalist, v. 13, p. 108-118. Maidment, D. R. ed., 1993, Handbook of hydrology: New York, McGraw-Hill Inc., p. 11.37-11.54. Mays, L. W., ed., 1996, Water resources handbook: New York, McGraw-Hill, p. 8.3-8.49. Natural Resources Conservation Service, National Water Quality Initiative: http://www.nrcs.usda.gov, accessed July 14, 2015. Mettee, M.F., O’Neil, P.E., Shepard, T.E., McGregor, S.W., and Henderson, W.P., Jr., 2002, A survey of protected fish species and species of uncommon occurrence in the Tennessee River drainage of north Alabama and northeast Mississippi: Geological Survey of Alabama Bulletin 171, 173 pages. Neves, R. J., 1999, Conservation and commerce: Management of freshwater mussel (Bivalvia: Unionoidea) resources in the United States: Malacologica, v. 4, no. 1(2), p. 461-474. O’Neil, P. E., and Meintzer, R. E., 1995, Quality assurance/quality control plans for the collection and analysis of water quality and aquatic biological samples: Alabama Geological Survey, unpublished report, 50 p. Owen, C. L., Bracken-Grissom, H., Stern, D., and Crandall, K. A., 2015, A synthetic phylogeny of freshwater crayfish: insights for conservation: Philosophical Transactions of the Royal Society B, 370, no. 20140009, 10 p., accessed July 12, 2015, from http://dx.doi.org/10.1098/rstb.2014.0009. Saylor, C. F., 1989, Sand Mountain-Guntersville Reservoir Biological Monitoring 1988 Index of Biotic Integrity: Tennessee Valley Authority, Resource Development, River Basins Operations, Water Resources, Water Quality Department, TVA/WR/WQ-89/4, 29 p. Shelton-Nix, E., ed., 2017, Alabama Wildlife, Volume 5: Tuscaloosa, University of Alabama Press, 372 p.

35 Taylor, C. A., 2002, Taxonomy and conservation of native crayfish stocks, in D.M. Holdich, ed., Biology of Freshwater Crayfish: Oxford, UK, Blackwell Science Ltd., p. 236–257. Taylor, C. A., Schuster, G. A., Cooper, J. E., Di Stefano, R. J., Eversole, A. G., Hamr, P., Hobbs, H. H., III., Robison, H. W., Skelton, C. E., and Thomas, R. E., 2007, A reassessment of the conservation status of crayfishes of the United States and Canada after 10+ years of increased awareness: Fisheries, v. 32, no. 8, p. 372–389. Tomlin, C. D. S., 2006, The Pesticide Manual: A World Compendium, 14th ed.: Surrey, UK, British Crop Protection Council. U.S. Department of Agriculture, 2012, National Agriculture Statistics Service, https://www.nass.usda.gov/Statistics_by_State/Alabama/Publications/AgStats/agstats, accessed June 22, 2016. U.S. Department of Agriculture, 2017, National Resources Conservation Service, https://www.nrcs.usda.gov/wps/portal/nrcs/detail/national/programs/initiatives/?cid=nrcs eprd1303651, accessed July 6, 2017. U.S. Environmental Protection Agency, 1973, Biological field and laboratory methods for measuring the quality of surface waters and effluents: Cincinnati, Ohio, United States Environmental Protection Agency, Office of Research and Development, EPA/670/4- 73/001. 1983, Methods for chemical analysis of water and wastes (revised edition): Cincinnati, Ohio, U.S. Environmental Protection Agency, Environmental Monitoring and Support Laboratory, EPA/600/4-79-020. 1988, Methods for the determination of organic compounds in drinking water: Cincinnati, Ohio, U.S. Environmental Protection Agency, Environmental Monitoring Systems Laboratory, EPA/600/4-88/039, 378 p. 1990, Methods for the determination of organic compounds in drinkingwater, supplement 1: Washington, D.C., U.S. Environmental Protection Agency, Office of Research and Development, EPA/600/4-90/020, 232 p. 1991, Methods for the determination of metals in environmental samples: Washington, D.C., U.S. Environmental Protection Agency, Office of Research and Development, EPA/600/4- 91/010, 293 p. 1998, Reregistration Eligibility Decision (RED), Alachlor: Washington, D.C., U.S. Government Printing Office, U.S. Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances (7508C), Office of Pesticide Programs,: EPA-738-F-98-018, 13 p. 2013, Aquatic life ambient water quality criteria for ammonia-freshwater: Washington, D.C., U.S. Environmental Protection Agency, Office of Water, EPA-822-R-13-001, 255 p. 2005, Reregistration Eligibility Decision (RED) 2,4-D: Washington, D.C., U.S. Government Printing Office, U.S. Environmental Protection Agency, Office of Prevention, Pesticides and Toxic Substances (7508C), Office of Pesticide Programs,: EPA- 738-R-05-002, xx p.

36 Warren, Jr., M. L., Burr, B. M., Walsh, S. J., Bart Jr., H. L., Cashner, R. C., Etnier, D. A., Freeman, B. J., Kuhajde, B. R., Mayden, R. L., Robison, H. W., and Starnes, W. D., 2000, Diversity, distribution, and conservation status of the native freshwater fishes of the southern United States: Fisheries, v. 25, no. 10, p. 7-31. Wershaw, R. L., Fishman, M. J., Grabbe, R. R., and Lowe, L. E., eds., 1987, Methods for the determination of organic substances in water and fluvial sediments: Techniques of water resources investigations of the U.S. Geological Survey, Book 5, Chapter A3, 80 p. World Health Organization, 1989, Environmental Aspects–2,4-Dichlorophenoxyacetic acid (2,4- D): Geneva, Switzerland, World Health Organization, International Programme on Chemical Safety, Environmental Health Criteria 84. .

37

38 APPENDIX A

WATER QUALITY DATA COLLECTED FROM SAMPLING SITES FOR THE 2015-2016 CAMBARUS CRACENS SURVEY CONDUCTED IN SCARHAM, SHORT, AND TOWN CREEK WATERSHEDS IN THE SAND MOUNTAIN AREA OF DEKALB AND MARSHALL COUNTIES, ALABAMA

39

40 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Alkalinity Watershed Ag Al mg/L as HCO3 CO3 CO2, Free Site Latitude Longitude Date Time Discharge area µg/L µg/L CaCO3 mg/L mg/L mg/L DEK04 34.3918 -86.0193 27-Apr-16 1545 24.54 149.00 <10 104 2 2 <1 1 DEK06 34.4178 -86.0405 27-Apr-16 1845 3.54 21.50 <10 <60 6 7 <1 4 DEK17 34.5731 -85.7508 27-Apr-16 1645 2.31 14.00 <10 <60 5 6 <1 2 DEK19 34.3308 -85.9778 27-Apr-16 1445 1.89 11.50 <10 <60 1 1 <1 <1 DEK20 34.6051 -85.7222 27-Apr-16 1745 0.64 3.89 <10 <60 3 4 <1 1 MAL03 34.3077 -86.1295 27-Apr-16 1045 0.01 0.04 <10 <60 7 9 <1 11 MAL05 34.2927 -86.1665 27-Apr-16 945 14.00 85.00 <10 <60 11 13 <1 208 MAL09 34.3638 -86.1243 27-Apr-16 1145 0.77 4.70 <10 89 10 12 <1 10 MAL10 34.3475 -86.1249 27-Apr-16 1245 1.30 7.88 <10 <60 7 9 <1 7 MAL13 34.2315 -86.1230 27-Apr-16 1345 6.00 36.40 <10 <60 9 11 <1 6 DEK04 34.3918 -86.0193 12-Oct-16 1145 0.12 149.00 <10 <60 26 32 <1 13 DEK08 34.4348 -86.0306 12-Oct-16 1045 0.01 15.40 <10 153 26 32 <1 13 DEK17 34.5731 -85.7508 12-Oct-16 930 0.01 14.00 <10 <60 42 51 <1 20 DEK19 34.3308 -85.9778 12-Oct-16 1300 0.01 11.50 <10 <60 209 254 <1 26 DEK22 34.6966 -85.6177 12-Oct-16 730 0.00 3.70 <10 <60 36 44 <1 140 MAL03 34.3077 -86.1295 12-Oct-16 1400 0.00 0.04 <10 <60 110 134 <1 11 MAL05 34.2927 -86.1665 12-Oct-16 1600 0.07 85.00 <10 <60 43 52 <1 5 MAL09 34.3638 -86.1243 12-Oct-16 1500 0.00 4.70 <10 <60 66 80 <1 5 MAL10 34.3475 -86.1249 12-Oct-16 1530 0.01 7.88 <10 <60 41 50 <1 6 MAL13 34.2315 -86.1230 12-Oct-16 1643 0.03 36.40 <10 <60 106 129 <1 21

Min <10 <60 1 1 <1 1 Med <10 <60 19 23 <1 10 Max <10 153 209 254 <1 208 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Total Residual Arsenic B Ba Be Br Ca Cd Cl Chlorine CN Co Conductivity Site µg/L µg/L µg/L µg/L mg/L mg/L µg/L mg/L mg/L mg/L µg/L µS/cm DEK04 0.80 15 63.3 <1 <0.02 6.1 <0.1 3.62 0.04 <0.003 <7 55 DEK06 <0.3 29 72.2 <1 <0.02 8.3 0.11 4.55 0.05 <0.003 <7 67 DEK17 0.31 13 52.1 <1 <0.02 7.3 <0.1 3.42 0.02 <0.003 <7 66 DEK19 0.81 20 86.0 <1 <0.02 5.5 <0.1 4.71 0.05 <0.003 <7 52 DEK20 <0.3 15 57.7 <1 <0.02 4.4 0.10 3.03 <0.02 <0.003 <7 38 MAL03 0.86 <10 64.8 <1 <0.02 6.6 <0.1 4.65 0.05 <0.003 <7 80 MAL05 1.00 11 62.3 <1 <0.02 6.3 <0.1 4.08 0.02 <0.003 <7 175 MAL09 0.88 <10 68.9 <1 <0.02 7.6 <0.1 5.55 0.04 <0.003 <7 73 MAL10 0.67 <10 64.0 <1 <0.02 6.9 0.12 4.98 0.03 <0.003 <7 65 MAL13 0.97 <10 46.9 <1 <0.02 6.9 <0.1 3.63 0.07 <0.003 <7 57 DEK04 1.02 14 80.5 <1 <0.02 8.7 <0.1 7.17 <0.02 <0.003 <7 77 DEK08 1.01 <10 99.5 <1 <0.02 10.4 <0.1 7.50 0.05 <0.003 <7 92 DEK17 <0.3 15 91.7 <1 0.05 23.7 <0.1 16.20 0.02 <0.003 <7 212 DEK19 1.93 <10 149.0 <1 1.27 74.8 <0.1 3.27 0.07 <0.003 <7 306 DEK22 1.95 <10 87.6 <1 <0.02 7.6 <0.1 2.58 0.02 <0.003 <7 78 MAL03 0.76 <10 149.0 <1 <0.02 36.7 <0.1 5.78 0.12 <0.003 <7 192 MAL05 0.74 <10 82.1 <1 <0.02 11.7 <0.1 5.23 0.09 <0.003 <7 90 MAL09 <0.3 <10 106.0 <1 <0.02 17.1 <0.1 6.71 0.21 <0.003 <7 125 MAL10 1.92 <10 114.0 <1 <0.02 16.1 <0.1 7.16 0.11 <0.003 <7 96 MAL13 0.67 21 159.0 <1 <0.02 32.0 <0.1 15.20 0.19 <0.003 <7 194

Min <0.3 <10 46.9 <1 <0.02 4.4 <0.1 2.58 <0.02 <0.003 <7 38 Med 0.81 <10 81.3 <1 <0.02 7.9 <0.1 4.85 0.05 <0.003 <7 79 Max 1.95 29 159.0 <1 1.27 74.8 0.12 16.20 0.21 <0.003 <7 306 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Dissolved Solids, Hardness Cr Cs Cu DO calculated F Fe mg/L as Hg K Li Mg Site µg/L µg/L µg/L mg/L mg/L mg/L µg/L CaCO3 µg/L mg/L µg/L mg/L DEK04 <0.3 <4 <8 10.6 32.1 <0.040 205.0 23.3 <0.005 2.27 <3 1.93 DEK06 <0.3 <4 <8 9.3 39.5 <0.040 184.0 28.2 <0.005 3.99 <3 1.82 DEK17 <0.3 <4 <8 9.4 42.0 0.044 289.0 31.3 <0.005 1.66 3.8 3.17 DEK19 <0.3 <4 <8 10.8 32.0 <0.040 161.0 19.5 <0.005 2.40 <3 1.41 DEK20 <0.3 <4 <8 9.2 27.8 <0.040 73.0 15.9 <0.005 1.29 3.3 1.22 MAL03 <0.3 <4 <8 9.2 39.7 <0.040 166.0 23.3 <0.005 2.81 <3 1.66 MAL05 <0.3 <4 <8 8.4 38.5 0.046 428.0 22.9 <0.005 2.26 <3 1.72 MAL09 <0.3 <4 <8 9.7 46.1 <0.040 215.0 27.4 <0.005 3.64 <3 2.06 MAL10 <0.3 <4 <8 10.5 40.2 <0.040 85.4 25.3 <0.005 3.22 <3 1.92 MAL13 <0.3 <4 <8 9.6 31.8 <0.040 669.0 24.9 <0.005 1.91 <3 1.85 DEK04 <0.3 <4 <8 6.9 54.1 0.384 142.0 38.8 0.0052 3.77 <3 4.12 DEK08 <0.3 <4 <8 4.2 61.1 0.371 1,770.0 40.0 0.0079 5.03 <3 3.37 DEK17 <0.3 <4 <8 7.5 155.8 0.365 201.0 101.8 <0.005 2.40 5.4 10.30 DEK19 <0.3 <4 <8 3.5 228.8 0.452 44.2 210.9 0.0051 4.06 <3 5.77 DEK22 <0.3 <4 <8 5.2 51.0 0.367 3,450.0 27.6 <0.005 4.16 <3 2.08 MAL03 <0.3 <4 <8 6.3 137.1 0.370 133.0 118.9 <0.005 5.57 <3 6.56 MAL05 <0.3 <4 <8 6.4 60.4 0.368 277.0 44.5 <0.005 3.65 <3 3.68 MAL09 <0.3 <4 <8 5.0 87.7 0.361 618.0 60.3 <0.005 4.39 <3 4.23 MAL10 <0.3 <4 <8 4.0 72.6 0.368 1,130.0 56.9 <0.005 4.38 3.1 4.02 MAL13 <0.3 <4 <8 3.2 139.1 0.379 138.0 102.4 <0.005 4.63 <3 5.41

Min <0.3 <4 <8 3.2 27.8 <0.040 44.2 15.9 <0.005 1.29 <3 1.22 Med <0.3 <4 <8 8.0 48.5 0.204 203.0 29.8 <0.005 3.65 <3 2.63 Max <0.3 <4 <8 10.8 228.8 0.452 3450.0 210.9 0.008 5.57 5.4 10.3 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Mn Mo Na Ni NH3_N TKN NO2_N NO2_NO2 NO3_N NO3_NO3 NOx_N NOx_NO3 Site µg/L µg/L mg/L µg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L mg/L DEK04 32.1 <20 2.14 <10 0.173 0.22 <0.02 <0.07 0.76 3.36 0.76 3.36 DEK06 36.5 <20 2.84 <10 0.151 0.29 <0.02 <0.07 1.10 4.87 1.10 4.87 DEK17 116 <20 2.39 <10 0.184 0.20 <0.02 <0.07 0.47 2.08 0.47 2.08 DEK19 13.1 <20 2.18 <10 0.195 0.21 <0.02 <0.07 1.05 4.65 1.05 4.65 DEK20 29.8 <20 2.00 <10 0.087 0.10 <0.02 <0.07 0.90 3.98 0.90 3.98 MAL03 22.0 <20 2.63 <10 0.201 0.81 <0.02 <0.07 1.02 4.52 1.02 4.52 MAL05 32.1 <20 2.26 <10 0.181 0.77 <0.02 <0.07 0.75 3.32 0.75 3.32 MAL09 29.4 <20 2.73 <10 0.166 1.89 <0.02 <0.07 1.57 6.95 1.57 6.95 MAL10 2.5 <20 2.53 <10 0.216 0.52 <0.02 <0.07 1.29 5.71 1.29 5.71 MAL13 43.6 <20 2.38 <10 0.155 0.19 <0.02 <0.07 0.34 1.51 0.34 1.51 DEK04 12 <20 3.44 <10 <0.05 0.75 <0.02 <0.07 0.96 4.25 0.96 4.25 DEK08 647 <20 4.75 <10 0.119 1.81 <0.02 <0.07 0.70 3.08 0.70 3.08 DEK17 177 <20 9.28 <10 <0.05 0.44 <0.02 <0.07 0.60 2.66 0.60 2.66 DEK19 690 <20 1.67 <10 0.751 1.43 <0.02 <0.07 0.48 2.12 0.48 2.12 DEK22 1,350 <20 1.02 <10 0.956 2.19 <0.02 <0.07 0.21 0.93 0.21 0.93 MAL03 212 <20 3.09 <10 0.067 0.74 0.078 0.26 0.39 1.73 0.47 2.07 MAL05 123 <20 3.88 <10 0.156 0.55 <0.02 <0.07 0.35 1.55 0.35 1.55 MAL09 641 <20 2.50 <10 0.118 0.43 <0.02 <0.07 0.74 3.28 0.74 3.28 MAL10 863 <20 4.07 <10 <0.05 0.61 <0.02 <0.07 0.77 3.41 0.77 3.41 MAL13 2,460 <20 6.82 <10 0.563 0.84 <0.02 <0.07 0.38 1.68 0.38 1.68

Min 2.5 <20 1.02 <10 <0.05 0.10 <0.02 <0.07 0.21 0.93 0.21 0.93 Med 79.8 <20 2.58 <10 0.161 0.58 <0.02 <0.07 0.75 3.30 0.75 3.30 Max 2460 <20 9.28 <10 0.956 2.19 0.078 0.26 1.57 6.95 1.57 6.95 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

Total Disolved P PO4_P Pb pH Rb Sb Se SiO2 Sn SO4 Sr Solids Site mg/L mg/L µg/L units µg/L µg/L µg/L mg/L µg/L mg/L µg/L mg/L DEK04 0.054 <0.03 3.51 6.7 10.5 <0.5 <0.8 2.62 <50 8.87 18.4 121 DEK06 0.038 <0.03 20.50 6.5 16.8 <0.5 <0.8 1.93 <50 7.61 20.3 125 DEK17 0.019 <0.03 5.47 6.6 8.3 <0.5 <0.8 5.08 <50 13.60 30.5 96 DEK19 0.038 <0.03 <0.4 6.6 9.3 <0.5 <0.8 4.55 <50 5.97 15.6 79 DEK20 0.011 <0.03 10.00 6.7 6.1 0.7 <0.8 5.83 <50 4.04 11.8 59 MAL03 0.040 <0.03 2.50 6.1 10.6 <0.5 <0.8 4.97 <50 7.30 18.8 59 MAL05 0.033 <0.03 2.92 5.0 8.4 <0.5 <0.8 7.16 <50 4.50 17.0 57 MAL09 0.045 <0.03 2.13 6.3 13.6 <0.5 <0.8 4.81 <50 6.64 24.1 79 MAL10 0.020 <0.03 1.75 6.3 12.5 <0.5 <0.8 4.32 <50 6.10 22.4 66 MAL13 0.043 <0.03 2.58 6.5 7.7 <0.5 <0.8 4.24 <50 3.36 15.6 54 DEK04 0.011 <0.03 2.42 6.6 6.0 <0.5 <0.8 0.39 <50 5.99 44.9 93 DEK08 0.226 <0.03 2.38 6.6 8.3 <0.5 <0.8 3.87 <50 5.17 46.8 108 DEK17 <0.010 <0.03 3.27 6.6 4.4 <0.5 <0.8 5.89 <50 59.7 126.0 184 DEK19 0.030 <0.03 1.77 7.2 6.1 <0.5 <0.8 5.74 <50 5.80 169.0 253 DEK22 0.106 <0.03 2.77 5.7 7.9 <0.5 <0.8 3.05 <50 4.12 34.6 61 MAL03 0.010 <0.03 2.00 7.3 7.4 <0.5 <0.8 1.14 <50 9.73 126.0 228 MAL05 <0.010 <0.03 2.28 7.2 5.1 <0.5 <0.8 0.83 <50 3.60 49.1 92 MAL09 <0.010 <0.03 1.88 7.4 8.3 <0.5 <0.8 5.23 <50 3.92 59.8 155 MAL10 0.046 <0.03 2.50 7.1 8.4 <0.5 <0.8 4.52 <50 2.84 65.4 150 MAL13 0.027 <0.03 2.24 7.0 11.3 <0.5 <0.8 5.50 <50 3.76 85.5 265

Min <0.010 <0.03 <0.4 5.0 4.4 <0.5 <0.8 0.39 <50 2.84 11.8 54 Med 0.032 <0.03 2.46 6.6 8.3 <0.5 <0.8 4.54 <50 5.89 32.6 95 Max 0.226 <0.03 20.5 7.4 16.8 0.7 <0.8 7.16 <50 59.7 169.0 265 Appendix A. Water quality data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama

TDS calculated Total Biochemic Total from Suspende al Oxygen Organic conductivity d Solids Temp Ti Tl Turbidity V Zn COD Demand Phenolics Carbon Site mg/L mg/L °C µg/L µg/L NTU µg/L µg/L mg/L mg/L µg/L mg/L DEK04 36 39 23 <4 <0.4 1.9 <4.0 20.8 93 1.0 9.3 3.75 DEK06 44 <4 21 <4 <0.4 1.9 <4.0 27.8 77 0.9 9.5 5.01 DEK17 43 142 22 <4 <0.4 4.6 <4.0 24.7 75 0.7 11.4 2.52 DEK19 34 <4 22 <4 <0.4 3.2 <4.0 36.8 65 0.8 11.2 3.24 DEK20 25 <4 20 <4 <0.4 1.5 <4.0 26.4 67 0.3 9.6 1.64 MAL03 52 <4 22 <4 <0.4 1.5 <4.0 10.3 97 0.5 <3 3.40 MAL05 114 <4 21 <4 <0.4 2.1 <4.0 20.3 141 0.6 <3 3.38 MAL09 47 5 23 <4 <0.4 2.5 <4.0 7.7 91 0.9 <3 3.81 MAL10 42 <4 22 <4 <0.4 2.3 <4.0 16.2 94 0.7 9.3 3.29 MAL13 37 9 22 <4 <0.4 3.3 <4.0 9.8 75 0.5 8.2 4.72 DEK04 50 <4 18 <4 <0.4 0.6 <4.0 55.9 87 0.9 <3 6.01 DEK08 60 44 15 <4 <0.4 29.4 <4.0 48.5 240 4.3 <3 3.64 DEK17 138 <4 14 <4 <0.4 4.2 <4.0 40.1 61 0.8 6.4 <1 DEK19 199 4 16 4.2 <0.4 7.4 <4.0 32.3 72 2.3 <3 <1 DEK22 51 545 12 <4 <0.4 82.0 <4.0 44.0 88 4.0 <3 7.46 MAL03 125 <4 19 <4 <0.4 1.7 <4.0 33.3 88 1.3 <3 1.56 MAL05 59 <4 18 <4 <0.4 1.8 <4.0 39.6 72 0.9 <3 1.30 MAL09 81 <4 17 <4 <0.4 6.0 <4.0 27.8 86 1.2 <3 <1 MAL10 62 <4 20 <4 <0.4 5.6 <4.0 29.8 110 2.9 <3 4.28 MAL13 126 4 18 <4 <0.4 15.9 <4.0 42.1 124 2.7 3.8 <1

Min 25 <4 12 <4 <0.4 0.60 <4.0 7.7 61 0.3 <3 <1 Med 52 <4 20 <4 <0.4 2.85 <4.0 28.8 88 0.9 <3 3.34 Max 199 545 23 4.2 <0.4 82.0 <4.0 55.9 240 4.3 11.4 7.46 47 APPENDIX B

PESTICIDE AND BACTERIA DATA COLLECTED FROM SAMPLING SITES FOR THE 2015-2016 CAMBARUS CRACENS SURVEY CONDUCTED IN SCARHAM, SHORT, AND TOWN CREEK WATERSHEDS IN THE SAND MOUNTAIN AREA OF DEKALB AND MARSHALL COUNTIES, ALABAMA

48 49 Appendix B. Pesticide and bacteria data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama Total Coliform Watershed Bacteria E. coli Discharge area cfu/100 cfu/100 Alachlor Atrazine 2,4-D Glyphosate Site No. Latitude Longitude Date (ft.3/s) (mi2) mL mL µg/L µg/L µg/L µg/L DEK01 34.2990 -86.0773 7-Dec-15 47.13 49 6,700 290.9 <0.042 0.048 <0.7 <0.05 DEK02 34.2944 -86.0966 7-Dec-15 50.40 52.4 6,630 275.5 <0.042 0.048 <0.7 <0.05 DEK03 34.3837 -86.1056 7-Dec-15 <.5 <0.5 27,550 2,419.6 <0.042 0.048 <0.7 <0.05 DEK04 34.3918 -86.0193 7-Dec-15 143.30 149 2,420 137.4 <0.042 <0.046 <0.7 <0.05 DEK05 34.4006 -86.0672 7-Dec-15 151.00 157 6,770 141.4 <0.042 0.063 <0.7 <0.05 DEK06 34.4178 -86.0405 7-Dec-15 20.68 21.5 5,830 198.9 <0.042 0.079 <0.7 <0.05 DEK07 34.4229 -86.0358 7-Dec-15 19.33 20.1 8,550 275.5 <0.042 0.079 <0.7 <0.05 DEK08 34.4348 -86.0306 7-Dec-15 14.81 15.4 9,060 90.9 <0.042 0.079 <0.7 <0.05 DEK09 34.4539 -86.0132 7-Dec-15 12.70 13.2 9,900 344.8 <0.042 0.090 <0.7 <0.05 DEK10 34.4472 -85.9799 7-Dec-15 10.19 10.6 10,760 260.3 <0.042 <0.046 <0.7 <0.05 DEK11 34.4774 -85.8095 7-Dec-15 84.54 87.9 5,830 95.9 <0.042 0.079 <0.7 <0.05 DEK12 34.5203 -85.7669 7-Dec-15 43.86 45.6 5,040 131.4 <0.042 <0.046 <0.7 <0.05 DEK13 34.5247 -85.7823 7-Dec-15 23.95 24.9 4,650 101.7 <0.042 <0.046 <0.7 <0.05 DEK14 34.5430 -85.7652 7-Dec-15 0.54 0.56 5,040 157.6 <0.042 0.090 <0.7 <0.05 DEK15 34.5512 -85.7607 7-Dec-15 18.47 19.2 4,040 95.9 <0.042 0.058 <0.7 <0.05 DEK16 34.5480 -85.7193 7-Dec-15 36.93 38.4 4,800 101.4 <0.042 0.145 <0.7 <0.05 DEK17 34.5731 -85.7508 7-Dec-15 13.46 14 4,730 613.1 <0.042 <0.046 <0.7 <0.05 DEK18 34.5850 -85.7404 7-Dec-15 6.88 7.15 5,730 178.5 <0.042 0.145 <0.7 <0.05 MAL01 34.2363 -86.1203 7-Dec-15 67.04 69.7 9,080 613.1 <0.042 0.134 <0.7 <0.05 MAL02 34.2984 -86.1162 7-Dec-15 52.51 54.6 5,860 275.5 <0.042 0.157 <0.7 <0.05 MAL03 34.3077 -86.1295 7-Dec-15 0.04 0.04 10,170 228.2 <0.042 0.090 <0.7 <0.05 MAL04 34.2588 -86.1240 7-Dec-15 69.05 71.8 9,080 365.4 <0.042 0.134 <0.7 <0.05 MAL05 34.2927 -86.1665 7-Dec-15 81.75 85 5,650 285.1 <0.042 0.145 <0.7 <0.05 MAL06 34.2976 -86.1793 7-Dec-15 4.70 4.89 9,080 44.0 <0.042 0.169 <0.7 <0.05 MAL07 34.3204 -86.2046 7-Dec-15 107.72 112 5,940 275.5 <0.042 0.145 <0.7 <0.05 MAL08 34.3267 -86.1615 7-Dec-15 29.43 30.6 4,730 218.7 <0.042 0.157 <0.7 <0.05 MAL09 34.3638 -86.1243 7-Dec-15 4.52 4.7 5,540 248.1 <0.042 0.193 <0.7 <0.05 MAL10 34.3475 -86.1249 7-Dec-15 7.58 7.88 4,350 172.3 <0.042 0.134 <0.7 <0.05 MAL11 34.3673 -86.1215 7-Dec-15 4.27 4.44 3,690 198.9 <0.042 0.169 <0.7 <0.05 Appendix B. Pesticide and bacteria data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama Total Coliform Watershed Bacteria E. coli Discharge area cfu/100 cfu/100 Alachlor Atrazine 2,4-D Glyphosate Site No. Latitude Longitude Date (ft.3/s) (mi2) mL mL µg/L µg/L µg/L µg/L MAL12 34.3787 -86.1137 7-Dec-15 2.89 3.01 12,360 1,046.2 <0.042 0.100 <0.7 <0.05 DEK04 34.3918 -86.0193 11-May-16 9.31 149 3,590.0 38.3 <0.042 <0.046 <0.7 <0.05 DEK05 34.4006 -86.0672 11-May-16 9.80 157 1,986.3 69.1 <0.042 <0.046 <0.7 <0.05 DEK06 34.4178 -86.0405 11-May-16 1.34 21.5 2,419.6 88.4 <0.042 <0.046 <0.7 <0.05 DEK15 34.5512 -85.7607 11-May-16 1.20 19.2 1,119.6 21.1 <0.042 <0.046 <0.7 <0.05 DEK19 34.3308 -85.9778 11-May-16 0.72 11.5 7,540.0 60.2 <0.042 <0.046 <0.7 <0.05 DEK20 34.6051 -85.7222 11-May-16 0.24 3.89 1,413.6 108.1 <0.042 <0.046 <0.7 <0.05 DEK21 34.6469 -85.6515 11-May-16 0.40 6.36 2,419.6 178.9 <0.042 <0.046 <0.7 <0.05 DEK22 34.6966 -85.6177 11-May-16 0.23 3.7 1,119.6 248.1 <0.042 <0.046 <0.7 <0.05 DEK23 34.6105 -85.6550 11-May-16 0.03 0.46 4,640.0 27.9 <0.042 <0.046 <0.7 <0.05 DEK24 34.4269 -85.8756 11-May-16 6.93 111 3,410.0 27.5 <0.042 <0.046 1.6 0.218 DEK25 34.3932 -85.9578 11-May-16 8.06 129 1,986.3 26.2 <0.042 <0.046 <0.7 <0.05 DEK26 34.3046 -85.9849 11-May-16 0.66 10.5 10,170.0 27.9 <0.042 0.051 <0.7 <0.05 DEK27 34.3419 -86.0083 11-May-16 0.04 0.67 6,310.0 365.4 <0.042 <0.046 <0.7 <0.05 DEK28 34.2388 -86.0760 11-May-16 1.79 28.6 8,360.0 166.4 <0.042 0.155 <0.7 <0.05 MAL01 34.2990 -86.0773 11-May-16 4.35 69.7 5,210.0 145.0 <0.042 <0.046 <0.7 <0.05 MAL03 34.3837 -86.1056 11-May-16 0.00 0.04 7,120.0 69.7 <0.042 0.2 <0.7 <0.05 MAL05 34.4006 -86.0672 11-May-16 5.31 85 5,290.0 30.1 <0.042 <0.046 <0.7 <0.05 MAL08 34.4348 -86.0306 11-May-16 1.91 30.6 4,790.0 27.8 <0.042 0.051 <0.7 0.172 MAL09 34.4539 -86.0132 11-May-16 0.29 4.7 57,940.0 235.9 <0.042 0.43 <0.7 <0.05 MAL10 34.4472 -85.9799 11-May-16 0.49 7.88 7,940.0 38.9 <0.042 0.34 <0.7 <0.05 DEK09 34.4539 -86.0132 8-Aug-16 0.17 13.2 14,390 39.3 <0.042 <0.046 <0.7 <0.05 DEK14 34.4539 -86.0132 8-Aug-16 0.01 0.56 173,290 3,500.0 <0.042 0.088 3.8 0.12 DEK16 34.5480 -85.7193 8-Aug-16 0.49 38.4 173,290 1,986.3 <0.042 0.046 <0.7 <0.05 DEK17 34.5731 -85.7508 8-Aug-16 0.18 14 22,240 151.5 <0.042 <0.046 <0.7 <0.05 DEK18 34.5850 -85.7404 8-Aug-16 0.09 7.15 38,730 5,560.0 <0.042 <0.046 <0.7 <0.05 DEK19 34.3308 -85.9778 8-Aug-16 0.15 11.5 3,010 59.4 <0.042 <0.046 <0.7 <0.05 DEK20 34.6051 -85.7222 8-Aug-16 0.05 3.89 2,310,000 4,710.0 <0.042 <0.046 <0.7 <0.05 DEK24 34.4269 -85.8756 8-Aug-16 1.40 111 6,440 65.7 <0.042 <0.046 <0.7 <0.05 Appendix B. Pesticide and bacteria data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama Total Coliform Watershed Bacteria E. coli Discharge area cfu/100 cfu/100 Alachlor Atrazine 2,4-D Glyphosate Site No. Latitude Longitude Date (ft.3/s) (mi2) mL mL µg/L µg/L µg/L µg/L DEK26 34.3046 -85.9849 8-Aug-16 0.13 10.5 32,550 2,419.6 <0.042 <0.046 <0.7 <0.05 DEK28 34.2388 -86.0760 8-Aug-16 0.36 28.6 22,240 435.2 <0.042 <0.046 <0.7 <0.05 DEK29 34.3053 -85.9941 8-Aug-16 0.33 26.1 3,990 <1 <0.042 0.057 <0.7 <0.05 MAL01 34.2363 -86.1203 8-Aug-16 0.88 69.7 8,860 34.5 <0.042 <0.046 <0.7 <0.05 MAL02 34.2984 -86.1162 8-Aug-16 0.69 54.6 6,450 613.1 <0.042 <0.046 <0.7 <0.05 MAL03 34.3077 -86.1295 8-Aug-16 0.00 0.04 155,310 2,419.6 <0.042 0.279 <0.7 <0.05 MAL04 34.2588 -86.1240 8-Aug-16 0.91 71.8 12,230 167.0 <0.042 0.050 <0.7 0.05 MAL05 34.2927 -86.1665 8-Aug-16 1.08 85 6,240 34.5 <0.042 0.072 <0.7 <0.05 MAL06 34.2976 -86.1793 8-Aug-16 0.06 4.89 27,230 81.6 <0.042 0.101 <0.7 0.06 MAL07 34.3204 -86.2046 8-Aug-16 1.42 112 21,430 261.3 <0.042 0.148 1.1 0.29 MAL08 34.3267 -86.1615 8-Aug-16 0.39 30.6 34,480 461.1 <0.042 0.289 <0.7 0.27 MAL09 34.3638 -86.1243 8-Aug-16 0.06 4.7 18,500 52.9 <0.042 0.213 <0.7 <0.05 MAL10 34.3475 -86.1249 8-Aug-16 0.10 7.88 14,670 52.9 <0.042 <0.046 <0.7 0.07 MAL11 34.3673 -86.1215 8-Aug-16 0.06 4.44 20,640 344.8 <0.042 0.116 <0.7 <0.05 MAL12 34.3787 -86.1137 8-Aug-16 0.04 3.01 7,710 2.0 <0.042 0.065 2.6 0.32 DEK09 34.4539 -86.0132 14-Sep-16 0.03 13.2 6,630 22.6 <0.042 <0.046 <0.7 <0.05 DEK14 34.5430 -85.7652 14-Sep-16 0.00 0.56 1,990 28.1 <0.042 <0.046 <0.7 <0.05 DEK16 34.5480 -85.7193 14-Sep-16 0.09 38.4 6,910 102.0 <0.042 <0.046 <0.7 <0.05 DEK17 34.5731 -85.7508 14-Sep-16 0.03 14 1,410 4.1 <0.042 <0.046 <0.7 <0.05 DEK18 34.5850 -85.7404 14-Sep-16 0.02 7.15 10,900 5.2 <0.042 <0.046 <0.7 0.051 DEK19 34.3308 -85.9778 14-Sep-16 0.03 11.5 22,500 101.0 <0.042 <0.046 17.4 <0.05 DEK20 34.6051 -85.7222 14-Sep-16 0.01 3.89 41,100 90.8 <0.042 <0.046 <0.7 0.121 DEK24 34.4269 -85.8756 14-Sep-16 0.27 111 2,420 12.0 <0.042 <0.046 <0.7 0.3 DEK26 34.3046 -85.9849 14-Sep-16 0.03 10.5 959,000 3,840.0 <0.042 <0.046 <0.7 0.064 DEK28 34.2388 -86.0760 14-Sep-16 0.07 28.6 19,400 9.6 <0.042 0.105 <0.7 <0.05 DEK29 34.3053 -85.9941 14-Sep-16 0.06 26.1 81,600 111.0 <0.042 <0.046 5.5 0.112 MAL01 34.2363 -86.1203 14-Sep-16 0.17 69.7 41,100 21.1 <0.042 <0.046 <0.7 0.112 MAL02 34.2984 -86.1162 14-Sep-16 0.13 54.6 11,500 8.5 <0.042 0.058 <0.7 <0.05 MAL03 34.3077 -86.1295 14-Sep-16 0.00 0.04 4,800 39.3 <0.042 0.111 <0.7 0.052 Appendix B. Pesticide and bacteria data collected from sampling sites for the 2015-2016 Cambarus cracens survey conducted in Scarham, Short, and Town Creek watersheds in the Sand Mountain area of DeKalb and Marshall Counties, Alabama Total Coliform Watershed Bacteria E. coli Discharge area cfu/100 cfu/100 Alachlor Atrazine 2,4-D Glyphosate Site No. Latitude Longitude Date (ft.3/s) (mi2) mL mL µg/L µg/L µg/L µg/L MAL04 34.2588 -86.1240 14-Sep-16 0.18 71.8 3,450 58.8 <0.042 0.135 <0.7 0.142 MAL05 34.2927 -86.1665 14-Sep-16 0.21 85 5,040 16.1 <0.042 <0.046 <0.7 <0.05 MAL06 34.2976 -86.1793 14-Sep-16 0.01 4.89 12,700 186.0 <0.042 0.239 <0.7 0.508 MAL07 34.3204 -86.2046 14-Sep-16 0.27 112 20,100 73.0 <0.042 0.046 <0.7 0.101 MAL08 34.3267 -86.1615 14-Sep-16 0.08 30.6 1,730 3.1 <0.042 0.230 <0.7 <0.05 MAL09 34.3638 -86.1243 14-Sep-16 0.01 4.7 29,100 13.1 <0.042 0.085 <0.7 <0.05 MAL10 34.3475 -86.1249 14-Sep-16 0.02 7.88 5,290 21.8 <0.042 0.115 <0.7 0.060 MAL11 34.3673 -86.1215 14-Sep-16 0.01 4.44 7,890 8.4 <0.042 0.135 <0.7 0.142 MAL12 34.3787 -86.1137 14-Sep-16 0.01 3.01 72,700 53.9 <0.042 <0.046 <0.7 <0.05 DEK04 34.3918 -86.0193 12-Oct-16 0.12 149 2,720.0 16.8 <0.042 0.079 <0.7 <0.05 DEK08 34.4348 -86.0306 12-Oct-16 0.01 15.4 325.5 1.0 <0.042 <0.046 <0.7 <0.05 DEK17 34.5731 -85.7508 12-Oct-16 0.01 14 2,090.0 292.4 <0.042 <0.046 <0.7 <0.05 DEK19 34.3308 -85.9778 12-Oct-16 0.01 11.5 1,732.9 16.8 <0.042 <0.046 <0.7 <0.05 DEK22 34.6966 -85.6177 12-Oct-16 0.00 3.7 6,170.0 161.6 <0.042 0.050 <0.7 <0.05 MAL03 34.3077 -86.1295 12-Oct-16 0.00 0.04 816.4 5.2 <0.042 0.122 <0.7 <0.05 MAL05 34.2927 -86.1665 12-Oct-16 0.07 85 2,419.6 4.1 <0.042 0.047 <0.7 <0.05 MAL09 34.3638 -86.1243 12-Oct-16 0.00 4.7 4,170.0 23.8 <0.042 <0.046 <0.7 <0.05 MAL10 34.3475 -86.1249 12-Oct-16 0.01 7.88 235.9 1.0 <0.042 0.123 <0.7 <0.05 MAL13 34.2315 -86.1230 12-Oct-16 0.03 36.4 1,970.0 184.2 <0.042 0.078 <0.7 <0.05

Min 236 1.0 <0.042 <0.046 <0.7 <0.05 Med 6,445 101 <0.042 0.049 <0.7 <0.05 Max 2,310,000 5,560 <0.042 0.430 17 0.508

GEOLOGICAL SURVEY OF ALABAMA 420 Hackberry Lane P.O. Box 869999 Tuscaloosa, Alabama 35486-6999 205/349-2852

Berry H. (Nick) Tew, Jr., State Geologist

A list of the printed publications by the Geological Survey of Alabama can be obtained from the Publications Office (205/247-3636) or through our web site at http://www.gsa.state.al.us.

E-mail: [email protected]

The Geological Survey of Alabama (GSA) makes every effort to collect, provide, and maintain accurate and complete information. However, data acquisition and research are ongoing activities of GSA, and interpretations may be revised as new data are acquired. Therefore, all information made available to the public by GSA should be viewed in that context. Neither the GSA nor any employee thereof makes any warranty, expressed or implied, or assumes any legal responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed in this report. Conclusions drawn or actions taken on the basis of these data and information are the sole responsibility of the user.

As a recipient of Federal financial assistance from the U.S. Department of the Interior, the GSA prohibits discrimination on the basis of race, color, national origin, age, or disability in its programs or activities. Discrimination on the basis of sex is prohibited in federally assisted GSA education programs. If anyone believes that he or she has been discriminated against in any of the GSA’s programs or activities, including its employment practices, the individual may contact the U.S. Geological Survey, U.S. Department of the Interior, Washington, D.C. 20240.

AN EQUAL OPPORTUNITY EMPLOYER

Serving Alabama since 1848