For Table and Figure references below, see the document "PredationAssay_statistical_analysis.pdf" in the Supplemental Files section.

Study Site Selection

We conducted our study across eight discrete seagrass meadows (hereafter referred to as landscapes) located in Back Sound, North Carolina (NC), USA (3442′ N to 3439′ N, 7637′ W to 7631′ W) (Fig. S1). All of our sampled landscapes were composed of a mixture of Back Sound's dominant seagrasses: eelgrass and shoal grass, Halodule wrightii (Ascherson 1868) (Yeager et al. 2016). Landscapes were chosen based upon available aerial imagery in Google Earth Pro as of February 19, 2017, and ground-truthed for changes in seasonal seagrass growth/senescence using summer, 2017, drone photography and ImageJ 1.x (Schneider et al. 2012). No discernable differences in landscape fragmentation states (e.g. total area, number of patches) were found between the two aerial imagery sources. All landscapes were relatively shallow (1-1.5 m depth at high tide), reasonably isolated from other seagrass beds (distance to nearest seagrass meadow = 112 17 m [mean standard error]) and were appropriately sized to encompass short-term (e.g., daily, monthly) movements of common seagrass-associated fauna in this system (Yeager et al. 2016). We identified similarly sized landscapes (25882 6592 m2) available in Back Sound by defining the minimum convex polygon surrounding the seagrass meadow, regardless of the total seagrass cover within the polygon. Among eight candidate landscapes of similar size, we defined four continuous landscapes and four fragmented landscapes based on the number of patches, the perimeter-to-area ratio, and the largest patch's percent cover of the total seagrass area (Table 1). Seagrass fragmentation is often naturally coupled with habitat loss (Wilcove et al. 1986), resulting in the mean seagrass area of our fragmented landscapes being nearly half that of our continuous landscapes (Table 1). Thus, our experiment was designed to examine the effects of fragmentation (i.e., the breaking apart of habitat concomitant with habitat loss) rather than fragmentation per se (i.e., the breaking apart of habitat without habitat loss; sensu Fahrig 2003).

Predation Assays

Relative predation (i.e., mortality) was measured using tethered juvenile blue crabs (Callinectes sapidus Rathbun) of carapace widths 10-40 mm. Tethering is commonly used to measure relative predation on juvenile blue crabs (Wilson et al. 1990; Hovel and Lipcius 2001). We note that tethering cannot be used to determine absolute predation rates, as tethered prey have restricted flee potential, generally raising the incidence of predation (Peterson and Black 1994). Still, when interpreted conservatively, tethering data can elucidate differences the relative directionality of environmental factors affecting prey survival (i.e., positive or negative effects). Juvenile blue crabs were chosen for tethering due to their economic and ecological importance to coastal regions (McCann et al. 2017) and because they have served as a model prey organism in several previous studies of related design (e.g., Hovel and Lipcius 2001, 2002; Mahoney et al. 2018).

Crab predation assays were run from June to July as this was the period during which we could obtain sufficient numbers of appropriately sized crabs for our experiment. All juvenile blue crabs were captured in seine nets from Oyster Creek, NC (3449'19"N, 7627'07"W). Crabs were glued (active ingredient cyanoacrylate) to 30-cm segments of 12-lbs test monofilament. We chose to use 30-cm segments of monofilament for tethers to allow crabs to exhibit natural burrowing behavior (Hovel and Fonseca 2005), mitigating some tethering artifacts of prey visibility (Peterson and Black 1994). Tethered crabs were attached to 60-cm long, 0.5-cm diameter, fiberglass stakes with attached floats for easy relocation. Once tethered, crabs were held overnight as a check for attachment integrity, and then deployed across our landscapes on the following day.

Twenty tethered crabs were deployed (stakes pushed completely into the sediment) in each continuous and fragmented landscape per predation assay date. Within each landscape, 10 tethered crabs were haphazardly placed within seagrass edges, defined as 30 cm (a tether length) from the seagrass-mudflat interface. The other 10 tethered crabs were haphazardly placed in seagrass interiors, defined as ≥1 m from the seagrass-mudflat interface. Only patches with a radius of 1 m or larger were used for tethers classified as 'interior'. However, patches with a radius of <1 m were used for a portion of our 'edge' tethers. All tethers were placed at least 1 m apart. We returned to landscapes at 1 h and 24 h to check crab status (live, eaten). All missing crabs were presumed eaten, as no crabs escaped tethers during the 24-h holding period. After 24 h, any remaining live crabs were removed from tethers and released. Crab tethering cycles were repeated four times in 2017 (June 9, June 14, July 5, and July 13). On July 13, only half of the continuous and fragmented landscapes were included in tethering assays due to a lack of crab availability. A total of 550 tethered juvenile blue crabs were deployed during our trials (see Table S1 for full sampling schedule and assay sample sizes).

Point measurements of water temperature (C) were taken in each landscape at the location and time of all tethering assays and faunal sampling using hand-held thermometers (Table S1). We chose temperature as our seasonality proxy (Fig. S2) because several other seasonally affected factors including faunal densities correlate with water temperature variability. Additionally, the measurement of temperature is easy, cheap, reliable, and comparable to previous studies.

Tether materials:
EcoStakes – tomato plant stakes
Locktite glue
12-lbs test monofilament fishing line
Pool noodles – cut into rounds for tether relocation floats
Hand-held digital thermometer- LYNCH Waterproof thermometer 39240
Hand-held refractometer-VEE GEE STX-3 Salinity 0-100%o
Hand-held Garmin GPSmap 78
Electronic calipers – INOX waterproof IP54