tors that also respond to ⌬ 9 -tetrahydrocannabinol (THC), the psychoactive component of marijuana ( 7 ). The genetic or pharmacological inactivation of FAAH results in substantial increases in brain concentrations of anandamide and other N -acyl ethanolamine (NAE) lipids and produces several CB1-dependent neurobehavioral effects in rodents, including anxiolysis ( 8, 9 ), anti-depression ( 10 ), and anti-nociception ( 6,11,12 ). Interestingly, these effects are not accompanied by the cognitive and motor dysfunctions associated with direct CB1 agonists such as THC. Taken together, these fi ndings indicate that FAAH is a key regulator of endocannabinoid activity in vivo and suggest further that the enzyme might represent a therapeutic target for the treatment of pain and other nervous system disorders ( 13,14 ).FAAH-disrupted mice have also shown some phenotypes that are not reversed by the administration of CB1 or CB2 antagonists ( 12,15,16 ), suggesting that anandamide and/ or other FAAH substrates possess bioactivities that extend beyond the endocannabinoid system. To more broadly explore the physiological substrate pool regulated by FAAH, our laboratory has analyzed FAAH( Ϫ / Ϫ ) mice using a metabolomics method based on untargeted LC-MS ( 17 ). This approach confi rmed known elevations in anandamide and other NAEs in brains and livers of FAAH( Ϫ / Ϫ ) mice and also uncovered a novel class of natural products regulated by FAAH, the N -acyl taurines (NATs). Subsequent in vitro studies showed that FAAH can hydrolyze NATs and that NATs are agonists of the transient receptor potential family of ion channels at concentrations approximately equal to or lower than those found in certain tissues from FAAH( Ϫ / Ϫ ) mice ( 17, 18 ). Fatty acid amide hydrolase (FAAH) is an integral membrane serine hydrolase that degrades a number of bioactive lipid amides, including the endocannabinoid anandamide ( N -arachidonoyl ethanolamine) ( 1, 2 ). Anandamide acts as an endogenous ligand for the CB1 and CB2 receptors ( 3-6 ), which are two G-protein-coupled recep- Abstract Fatty acid amide hydrolase (FAAH) regulates amidated lipid transmitters, including the endocannabinoid anandamide and its N -acyl ethanolamine (NAE) congeners and transient receptor potential channel agonists N -acyl taurines (NATs). Using both the FAAH inhibitor PF-3845 and FAAH( ؊ /؊

Infectious diseases play an important role in wildlife population dynamics by altering individual fitness, but detecting disease-driven natural selection in free-ranging populations is difficult due to complex disease–host relationships. Chronic wasting disease (CWD) is a fatal infectious prion disease in cervids for which mutations in a single gene have been mechanistically linked to disease outcomes, providing a rare opportunity to study disease-driven selection in wildlife. In Wyoming, USA, CWD has gradually spread across mule deer ( Odocoileus hemionus ) populations, producing natural variation in disease history to evaluate selection pressure. We used spatial variation and a novel temporal comparison to investigate the relationship between CWD and a mutation at codon 225 of the mule deer prion protein gene that slows disease progression. We found that individuals with the ‘slow’ 225F allele were less likely to test positive for CWD, and the 225F allele was more common in herds exposed to CWD longer. We also found that in the past 2 decades, the 225F allele frequency increased more in herds with higher CWD prevalence. This study expanded on previous research by analysing spatio-temporal patterns of individual and herd-based disease data to present multiple lines of evidence for disease-driven selection in free-ranging wildlife.

1Advances in DNA sequencing have made it feasible to gather genomic data for non-model 2 organisms and large sets of individuals, often using methods for sequencing subsets of the 3 genome. Several of these methods sequence DNA associated with endonuclease restriction 4 sites (various RAD and GBS methods). For use in taxa without a reference genome, these 5 methods rely on de novo assembly of fragments in the sequencing library. Many of the soft-6 ware options available for this application were originally developed for other assembly types 7 and we do not know their accuracy for reduced representation libraries. To address this im-8 portant knowledge gap, we simulated data from the Arabidopsis thaliana and Homo sapiens 9 genomes and compared de novo assemblies by six software programs that are commonly 10 used or promising for this purpose (ABySS, CD-HIT, Stacks, Stacks2, Velvet and VSEARCH). 11We simulated different mutation rates and types of mutations, and then applied the six 12 assemblers to the simulated datasets, varying assembly parameters. We found substantial 13 variation in software performance across simulations and parameter settings. ABySS failed 14 to recover any true genome fragments, and Velvet and VSEARCH performed poorly for most 15 simulations. Stacks and Stacks2 produced accurate assemblies of simulations containing 16 SNPs, but the addition of insertion and deletion mutations decreased their performance. 17CD-HIT was the only assembler that consistently recovered a high proportion of true genome 18 fragments. Here, we demonstrate the substantial difference in the accuracy of assemblies 19 from different software programs and the importance of comparing assemblies that result 20 from different parameter settings. 21

Understanding reproductive behavior of sensitive species is crucial for their conservation. The Delta Smelt Hypomesus transpacificus is a federally threatened, state‐endangered fish whose reproductive behavior is poorly understood. We used genetic techniques to investigate the spawning behavior of cultured Delta Smelt in a conservation hatchery. We conducted a natural tank‐spawning experiment in a total of four separate tanks during two spawning seasons. Delta Smelt were allowed to spawn in order to investigate spawning patterns using genetic parentage analysis of larvae produced. In total, 2,474 larvae were assigned two parents with >80% likelihood. Of the adults that had larvae assigned to them, males spawned on average 2.8 times and females 1.7 times. The mean number of larvae produced by females was 40.7, while males produced a mean number of 19.2 larvae during a single spawning season. Genetic diversity was reduced from the parent population to the offspring population, as indicated by a small but significant reduction in heterozygosity. Finally, we found no evidence that Delta Smelt preferred to mate with unrelated individuals.

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Preserving connectivity in the core of a species’ range is crucial for long-term persistence. However, a combination of ecological characteristics, social behavior, and landscape features can reduce connectivity among wildlife populations and lead to genetic structure. Pronghorn (Antilocapra americana), for example, exhibit fluctuating herd dynamics and variable seasonal migration strategies, but GPS tracking studies show that landscape features such as highways impede their movements, leading to conflicting hypotheses about expected levels of genetic structure. Given that pronghorn populations declined significantly in the early 1900s, have only partially recovered, and are experiencing modern threats from landscape modification, conserving connectivity among populations is important for their long-term persistence in North America. To assess the genetic structure and diversity of pronghorn in the core of their range, we genotyped 4,949 genome-wide single-nucleotide polymorphisms and 11 microsatellites from 398 individuals throughout the state of Wyoming. We found no evidence of genetic subdivision and minimal evidence of isolation by distance despite a range that spans hundreds of kilometers, multiple mountain ranges, and three interstate highways. In addition, a rare variant analysis using putatively recent mutations found no genetic division between pronghorn on either side of a major highway corridor. Although we found no evidence that barriers to daily and seasonal movements of pronghorn impede gene flow, we suggest periodic monitoring of genetic structure and diversity as a part of management strategies to identify changes in connectivity.

Advances in DNA sequencing have made it feasible to gather genomic data for non‐model organisms and large sets of individuals, often using methods for sequencing subsets of the genome. Several of these methods sequence DNA associated with endonuclease restriction sites (various RAD and GBS methods). For use in taxa without a reference genome, these methods rely on de novo assembly of fragments in the sequencing library. Many of the software options available for this application were originally developed for other assembly types and we do not know their accuracy for reduced representation libraries. To address this important knowledge gap, we simulated data from the Arabidopsis thaliana and Homo sapiens genomes and compared de novo assemblies by six software programs that are commonly used or promising for this purpose (ABySS, CD‐HIT, Stacks, Stacks2, Velvet and VSEARCH). We simulated different mutation rates and types of mutations, and then applied the six assemblers to the simulated data sets, varying assembly parameters. We found substantial variation in software performance across simulations and parameter settings. ABySS failed to recover any true genome fragments, and Velvet and VSEARCH performed poorly for most simulations. Stacks and Stacks2 produced accurate assemblies of simulations containing SNPs, but the addition of insertion and deletion mutations decreased their performance. CD‐HIT was the only assembler that consistently recovered a high proportion of true genome fragments. Here, we demonstrate the substantial difference in the accuracy of assemblies from different software programs and the importance of comparing assemblies that result from different parameter settings.