2, Appendix). The most dramatic decline, in both distribution and numbers, is in the Cypress Creek system (Fig. 2). Sites with positive detection have decreased with each successive sampling period.
Most notably, Slackwater Darter is now absent from the North Fork, Cypress Creek system. Although numbers of specimens are difficult to compare due to variable effort, studies from the 1970s reported 65 specimens from Selleckchem PLX3397 Lindsey Creek, while only 11 were collected in 1992–94; 10 were collected from Dulin Branch in the 1970s and 25 were collected in 1992–94; 19 were collected from Middle Cypress Creek and 53 were collected in 1992–94 (McGregor and Shepard 1995). Slackwater Darter was absent from other locations in 1992–94 and in the current study. Repeated sampling P005091 in vitro of the Middle Cypress Creek site during the breeding season (January to early March) (site 25, Figs. 1, 2) suggests a decline in numbers of Slackwater Darter collected over time (Fig. 3). Average, effort-adjusted numbers were: 109 in 2001 (n = 3 samples), 40 in 2002 (n = 2 samples), 21 in 2006 (n = 2), 25 in 2007 (n = 1), 6 in 2012 (n = 1) and 5 in 2013 (n = 1). Collections made in the seepage
area and CAL-101 supplier adjacent stream at different times of the year (February, March, July and August) indicate that the darters reside in both areas throughout the year. Fig. 3 Numbers of Etheostoma boschungi collected in Middle Cypress Creek (site 25) over time (2001–02, 2007–08, 2012–13), standardized for a 1 h effort Data on bank height ratio (BHR), taken at selected historical breeding sites, suggests a relationship between a low ratio, indicating probable connection between the stream and the floodplain, and a high ratio, unlikely
to maintain a connection to the floodplain during high water (Table 2). Sites with extant populations of Slackwater Darter had bank height ratios less than 2, while those where Slackwater Darter have not been recently detected had bank height ratios of 2.3–8.4. (mean BHR extant sites = 1.22, SD = 0.28; mean BHR extirpated sites = 4.95, SD = 2.4; F = 12.82, p = 0.007, t test). Table 2 Bank height ratios (BHR) measured in 2007 at selected historical and current sites of positive detection for Etheostoma boschungi, as a measure L-NAME HCl of current channel connectivity Site BHR Year last detected Lindsey, 4 6.0 1974 Lindsey, 7 4.0 1979 Natchez Trace, 20 1.0 2010 N Fork, 11 8.4 1979 Cemetery Branch, 10 2.3 1979 Elijah Branch, 12 6.6 1979 Middle Cypress, 25 1.3 2013 Brier Fork, 50a 2.4 1994 Brier Fork, 51 1.0 2007 Little Shoal, 34 1.6 2002 Positive versus negative detection in 2000s, F = 12.82, p = 0.007, t-test aSeepage area converted to a farm pond post 1995 Discussion These results suggest at least a 45 % historical range reduction of Slackwater Darter in approximately 15 years. In addition, the species had not been detected from a major portion of its range in the Cypress Creek system from the 1970 to the 1990s, and was not detected during this study.