2b) The dart was buoyant in water and floated with the dart tail

2b). The dart was buoyant in water and floated with the dart tail upright. We used a collapsible 1.2–3.7 m long pool net to retrieve darts in the water from the helicopter. We used tree check details marking paint or livestock marking solution (LA-CO Industries, Inc., Elk Grove, IL) for marking bears with this dart. We used an MK24C 0.745 projector (Paxarms N.Z. Ltd.) to fire the PX darts. For all dart types, we cleaned darts with soap and boiling water and used a 10% bleach solution as a disinfectant. When biopsy darting polar bears, we attempted to fire darts perpendicular to the body around the upper shoulder, similar to immobilization darting (Stirling et al. 1989). This approach

was used to help ensure darts would immediately bounce out from the large muscle upon impact. We typically darted http://www.selleckchem.com/products/acalabrutinib.html bears when they were approximately 3–6 m below the helicopter. Upon recovery of darts, we examined whether tissue samples had been collected and if not, we re-darted individuals when feasible. We examined the amount of time required to dart bears using the time at which bears were first observed

to the time the helicopter landed to recover fired darts. We weighed entire samples obtained in spring 2011 and September 2011. In August and September 2011, we separated adipose tissue from hair and skin and only submitted the hair and skin portion of the sample for genotyping. We air dried all genetic samples prior to DNA extraction. DNA was extracted from tissue samples using QIAGEN DNeasy Tissue Kits

according to manufacturer’s instructions by Wildlife Genetics International (WGI) Inc. (Nelson, British Columbia, Canada). WGI amplified DNA extracts at 20 microsatellite loci and the ZFX/ZFY sex identification marker (Aasen and Medrano 1990) using methods and primers as described in detail by Paetkau (2003) and Kendall et al. (2009). We considered genotyping successful if the DNA extract amplified at the full suite of microsatellite selleck kinase inhibitor loci and ZFX/ZFY. We extracted lipids from adipose, derivatized fatty acids to their fatty acid methyl ester (FAME) analogues using the Hilditch reagent, and quantified individual FAMEs by gas chromatography with flame ionization detection (Budge et al. 2006). We considered fatty acid analysis of the remote biopsies successful if we were able to quantify all fatty acids routinely determined in larger biopsies from captured or harvested polar bear samples (Thiemann et al. 2008; McKinney et al. 2009, 2011). We tested for normality in data sets using Shapiro-Wilk tests in the R programming language (http://cran.r-project.org/). We tested for differences in the mean wet weight of samples (the entire sample: hair, skin, and adipose tissue), mean adipose weight of samples, and adipose percent lipid content of samples obtained using the PC 5 mm heads, PX 5 mm heads, and PX 7 mm heads.

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