The Brook Floater (Alasmidonta varicosa) is a small (<100 mm), stream dwelling freshwater mussel (Family: Unionidae) from Atlantic Slope drainages in the eastern U.S. (Nedeau 2008). Brook Floater have dramatically contracted in distribution over recent decades, and there is limited evidence of recruitment in most locations, despite minimal effort to document population status (Wicklow et al., 2017). Brook Floater is listed as a Species of Greatest Conservation Need (SGCN) throughout its range in the United States (state-listed as imperiled or critically imperiled in all 15 states), has been extirpated from two states (Rhode Island and Delaware) and was recently petitioned for Federal listing in 2011 (Wicklow et al. 2017). Currently, there is a U.S. Fish and Wildlife (USFWS) Species Status Assessment underway to determine if federal listing under the Endangered Species Act is warranted. Brook Floater is also listed as a species of special concern in Canada, the northern extent of its range. In 2016, a state wildlife grant was awarded to develop range-wide conservation initiatives and strategies, including the development of rapid assessment and long-term monitoring techniques, in addition to developing conservation strategies to improve its probability of persistence in the future. The purpose of this protocol is to describe and facilitate a rapid approach to estimating Brook Floater occupancy to better understand the factors that influence Brook Floater distribution. Occupancy estimation approaches allow for estimation of species occupancy (; percent area occupied) within some scale of interest (for our purposes, the watershed), while simultaneously estimating species detection probability (p; the probability of finding an organism, if present). Occupancy estimation has been used with many wildlife taxa and is essential for understanding the presence or absence of wildlife in a particular area while accounting for imperfect detection (i.e., p<1; MacKenzie et al. 2004, Shea et al. 2013, Wisniewski et al. 2013, Pandolfo et al. 2016, MacKenzie 2016). This approach does not rely on existing information about species presence or absence to select sites. Occupancy estimation operates on a robust probabilistic framework of randomly selected sites to infer what proportion of sites are occupied. Occupancy estimation also incorporates imperfect detection (p <1; i.e., animals are cryptic and elusive; observers have varying experience searching, etc.; MacKenzie et al. 2003). For example, two mussel species that occupy a site might have two very different detection histories, as determined by revisiting a site and using the same methods on repeated visits to find both species. See hypothetical results here: Visits 1 2 3 4 5 Mussel species A 1 1 1 0 1 Mussel species B 0 0 0 1 0 (1 = detected, 0=not detected) Both of these mussel species occupy this site, yet Mussel A was detected in 4 out of 5 visits (high p) and Mussel B was detected in 1 out of 5 visits (low p) with the methods used to survey this site. Covariates may explain differences in detection between species or visits. Organisms may be: 1) present and not observed, 2) present and unavailable for capture (i.e., buried in sediment), or 3) not present at the site. Occupancy estimation uses repeated visits of randomly selected sites to build species detection histories (i.e., 1, 0, 1) to simultaneously estimate occupancy () and p. Typically, repeated visits are discrete sampling events and are more time consuming because each site requires >3 separate visits. In our rapid assessment protocol, we use multiple independent observers searching longitudinal lanes to estimate detection in a single site visit as opposed to multiple discrete visits. Below are hypothetical results of occupancy by observer: Independent Observers 1 2 3 4 5 Mussel species A 1 1 0 1 1 Mussel species B 1 0 0 1 0 (1 = detected, 0=not detected) Objectives: The objectives of this rapid assessment survey approach are to guide collection of data that can be used to: A. Estimate the occupancy of Brook Floater within watersheds. B. Estimate the effects of reach- and watershed-scale habitat features on Brook Floater occurrence. C. Understand how survey covariates (e.g., surveyor experience) influence detection of Brook Floater. While this protocol explicitly targets collection of Brook Floater, it is likely that the methods can be adapted for occupancy surveys of other stream-dwelling freshwater mussel species.