e. taken from the first GPS location; Tambling et al., 2012). Faeces were washed using water through a metal sieve (1.5-mm mesh), leaving only undigested prey remains – predominantly hair and bone fragments – and allowed to dry naturally. Undigested hair was separated from other remains and cleaned using equal measures of
alcohol p38 MAPK phosphorylation and sulphuric ether. Hair samples (≥30 individual strands) were randomly selected and analysed to identify prey species using hair cuticle scale patterns and cross sections (Mukherjee, Goyal & Chellam, 1994). Faecal samples containing the same species and located at the same GPS cluster were combined to avoid over-representation of prey items from multiple samples (Tambling et al., 2012). GPS cluster-located faecal samples were collected infrequently (median = every 11 days per leopard) and never in close proximity to another GPS cluster. Therefore, we assumed that the same prey individual was not represented in more than one faecal sample, provided it was not part of the same cluster. Three predation datasets were collected during MLN8237 cell line the study: (1) dietary estimates from GPS-located carcasses;
(2) dietary estimates from faecal samples collected at GPS-located clusters with and without located kills; (3) dietary estimates from a combination of faecal samples collected from GPS-located clusters with and without kills and opportunistically while traversing leopard home ranges. Datasets (1) and (2) are linked through time and space, as the collection of faecal samples and carcass observations occur chronologically. Therefore, these data can be
used in combination to form a detailed history of leopard feeding activity to better understand leopard feeding ecology (Martins et al., 2011). Faecal samples collected at GPS cluster sites could be a product of a kill located at that present feeding site, at a previous feeding site or at a feeding site undetected by the GPS cluster method (i.e. a missed kill). To determine which category each faecal sample belongs to, the average transit times of prey through the gut of leopards are required. Unfortunately, estimates of leopard gut transit times are not available, so we followed the procedure described RNA Synthesis inhibitor by Tambling et al. (2012), explained below. Based on cheetah digestion rates (48–111 h; Marker et al., 2003), we assigned two extreme gut transit times (minimum = 2 days and maximum = 5 days) for leopards. Faeces produced within the gut transit window of a leopard at a kill site are expected to contain the remains of the carcass found at that kill site. Faecal samples found outside of these transit limits and/or consisting of species other than the carcass found were considered to represent missed feeding events (Fig. 1). We calculated the number of missed feeding events for each prey species at both minimum and maximum gut transit times.