Using DNA to Identify the Source of Invasive Mongooses, \u3ci\u3eHerpestes auropunctatus\u3c/i\u3e (Carnivora: Herpestidae) Captured on Kaua‘i, Hawaiian Islands

Two small Indian mongooses (Herpestes auropunctatus) were live-captured in 2012 at separate locations on the Hawaiian Island of Kaua\u27i, which was previously considered to be free of this invasive species. Genotypes from these two individuals were compared to genotypes of H. auropunctatus from the islands of Hawai\u27i (n =39), O\u27ahu (n =91), Maui (n = 39), and Moloka\u27i (n = 19) to determine the island of origin of the Kaua\u27i individuals. Genotypes were generated from each individual using five microsatellite loci. Genetic clustering was estimated by Bayesian inference of spatial clustering of individuals and clustering of groups of individuals. Both analyses separated the samples into three distinct genetic clusters (K = 3). Kaua\u27i individuals consistently formed a single cluster with individuals from O\u27ahu, whereas Hawai\u27i and Maui formed a second cluster, and Moloka\u27i was the third cluster. Thus, we conclude that the origin of two H. auropunctatus captured on Kaua\u27I was O\u27ahu. All three genetic clusters showed evidence of transportation of mongooses between islands, indicating that sampled islands in the archipelago are capable of acting as both donors and receivers of mongooses

EVIDENCE OF TWO COCIRCULATING CANINE DISTEMPER VIRUS STRAINS IN MESOCARNIVORES FROM NORTHERN COLORADO, USA

Canine distemper virus (CDV) is a highly contagious pathogen that principally infects wildlife and domestic carnivores. Peridomestic species such as raccoons (Procyon lotor) experience outbreaks with high mortality. Clinical signs of infection include anorexia, fever, respiratory infection, and neurologic complications. Although not zoonotic, CDV poses a high risk to unvaccinated domestic animals and the conservation of endangered species. During 2013–16, we opportunistically collected wild and domestic carnivore specimens through a rabies surveillance program in northern Colorado, US. Brainstem and cerebellar tissue samples were independently tested for rabies and CDV by fluorescent antibody test. We tested a total of 478 animals for CDV, comprised of 10 wild and domestic carnivore species. A total of 15% (72/478) of all animals sampled tested positive for CDV, consisting of 24% (71/300) of raccoons and 4% (1/26) of coyotes (Canis latrans), but coinfection with rabies virus was not observed among CDV-positive animals. We extracted RNA from positive tissues, and a reverse-transcription PCR was used to create complementary DNA. We amplified and sequenced the hemagglutinin gene from 60 CDV-positive tissues, and a median joining network and maximum likelihood phylogenetic tree revealed two major lineages among samples. Phylogenetic analysis indicated that our sequences were most similar to the America-2 (n=55) and the America-3 (n=5) CDV lineages circulating in North America. Our results indicated two distinct and distantly related clades of CDV overlapping geographically and temporally among raccoon populations in northern Colorado

DNA Persistence in Predator Saliva from Multiple Species and Methods for Optimal Recovery from Depredated Carcasses

Molecular forensics is an important component of wildlife research and management. Using DNA from noninvasive samples collected at predation sites, we can identify predator species and obtain individual genotypes, improving our understanding of predator–prey dynamics and impacts of predators on livestock and endangered species. To improve sample collection strategies, we tested two sample collection methods and estimated degradation rates of predator DNA on the carcasses of multiple prey species. We fed carcasses of calves (Bos taurus) and lambs (Ovis aires) to three captive predator species: wolves (Canis lupus), coyotes (C. latrans), and mountain lions (Puma concolor). We swabbed the carcass in the field, as well as removed a piece of hide from the carcasses and then swabbed it in the laboratory. We swabbed all tissue samples through time and attempted to identify the predator involved in the depredation using salivary DNA. We found the most successful approach for yielding viable salivary DNA was removing hide from the prey and swabbing it in the laboratory. As expected, genotyping error increased through time and our ability to obtain complete genotypes decreased over time, the latter falling below 50% after 24 h. We provide guidelines for sampling salivary DNA from tissues of depredated carcasses for maximum probability of detection

Using DNA to Identify the Source of Invasive Mongooses, \u3ci\u3eHerpestes auropunctatus\u3c/i\u3e (Carnivora: Herpestidae) Captured on Kaua‘i, Hawaiian Islands

Two small Indian mongooses (Herpestes auropunctatus) were live-captured in 2012 at separate locations on the Hawaiian Island of Kaua\u27i, which was previously considered to be free of this invasive species. Genotypes from these two individuals were compared to genotypes of H. auropunctatus from the islands of Hawai\u27i (n =39), O\u27ahu (n =91), Maui (n = 39), and Moloka\u27i (n = 19) to determine the island of origin of the Kaua\u27i individuals. Genotypes were generated from each individual using five microsatellite loci. Genetic clustering was estimated by Bayesian inference of spatial clustering of individuals and clustering of groups of individuals. Both analyses separated the samples into three distinct genetic clusters (K = 3). Kaua\u27i individuals consistently formed a single cluster with individuals from O\u27ahu, whereas Hawai\u27i and Maui formed a second cluster, and Moloka\u27i was the third cluster. Thus, we conclude that the origin of two H. auropunctatus captured on Kaua\u27I was O\u27ahu. All three genetic clusters showed evidence of transportation of mongooses between islands, indicating that sampled islands in the archipelago are capable of acting as both donors and receivers of mongooses

EVIDENCE OF TWO COCIRCULATING CANINE DISTEMPER VIRUS STRAINS IN MESOCARNIVORES FROM NORTHERN COLORADO, USA

Canine distemper virus (CDV) is a highly contagious pathogen that principally infects wildlife and domestic carnivores. Peridomestic species such as raccoons (Procyon lotor) experience outbreaks with high mortality. Clinical signs of infection include anorexia, fever, respiratory infection, and neurologic complications. Although not zoonotic, CDV poses a high risk to unvaccinated domestic animals and the conservation of endangered species. During 2013–16, we opportunistically collected wild and domestic carnivore specimens through a rabies surveillance program in northern Colorado, US. Brainstem and cerebellar tissue samples were independently tested for rabies and CDV by fluorescent antibody test. We tested a total of 478 animals for CDV, comprised of 10 wild and domestic carnivore species. A total of 15% (72/478) of all animals sampled tested positive for CDV, consisting of 24% (71/300) of raccoons and 4% (1/26) of coyotes (Canis latrans), but coinfection with rabies virus was not observed among CDV-positive animals. We extracted RNA from positive tissues, and a reverse-transcription PCR was used to create complementary DNA. We amplified and sequenced the hemagglutinin gene from 60 CDV-positive tissues, and a median joining network and maximum likelihood phylogenetic tree revealed two major lineages among samples. Phylogenetic analysis indicated that our sequences were most similar to the America-2 (n=55) and the America-3 (n=5) CDV lineages circulating in North America. Our results indicated two distinct and distantly related clades of CDV overlapping geographically and temporally among raccoon populations in northern Colorado

Systematics, Distribution, and Conservation Status of Dice\u27s Cottontail, Sylvilagus Dicei Harris, 1932 (mammalia, Lagomorpha, Leporidae), in Central America

Dice’s cottontail, Sylvilagus dicei, only was recognized as a valid species in 1981 after having been subsumed in synonymy with S. brasiliensis and S. gabbi, which latter also at times had been synonymized with an ill-defined and broadly distributed S. brasiliensis. Phylogenetic relationships of S. dicei remain poorly resolved, as does the distribution of the species. We collected new specimens of S. dicei and carried out phylogenetic analyses of combined cytochrome-b and 12S rRNA mitochondrial loci. Molecular data confirm the taxonomic status of S. dicei as a species and place it basal to the South American S. brasiliensis radiation; the successive sister taxon to this clade is Gabb’s cottontail, S. gabbi. Time-calibrated analyses support simultaneous internal diversification events in S. dicei and S. gabbi ∼250 KYA. The new specimens, together with literature and museum records including the holotype, enabled us to more accurately map the distribution of S. dicei. Specimens are restricted to forested uplands in Costa Rica and western Panama. Although the total altitudinally suitable area for the species is 10,313 km2, removal of urban and agricultural areas not suited for its habitation result in a fragmented range of 9,115 km2. Conservation threats include high predation loads from the invasive coyote, Canis latrans, urbanization, agriculture, and hunting