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Previous studies have examined germ-line mutations to infer the processes that generate and maintain variability in microsatellite loci. Few studies, however, have examined patterns to infer processes that act on microsatellite loci over evolutionary time. Here, we examine changes in 8 dinucleotide loci across the adaptive radiation of Hawaiian honeycreepers. The loci were found to be highly variable across the radiation, and we did not detect ascertainment bias with respect to allelic diversity or allele size ranges. In examining patterns at the sequence level, we found that changes in flanking regions, repeat motifs, or repeat interruptions were often shared between closely related species and may be phylogenetically informative. Genetic distance measures based on microsatellites were strongly correlated with those based on mitochondrial DNA (mtDNA) sequences as well as with divergence time up to 3 My. Phylogenetic inferences based on microsatellite genetic distances consistently recovered 2 of the 4 honeycreeper clades observed in a tree based on mtDNA sequences but differed from the mtDNA tree in the relationships among clades. Our results confirm that microsatellite loci may be conserved over evolutionary time, making them useful in population-level studies of species that diverged from the species in which they were characterized as long as 5 Ma. Despite this, we found that their use in phylogenetic inference was limited to closely related honeycreeper species.

Recent genetic results support the recognition of two African elephant species: Loxodonta africana, the savannah elephant, and Loxodonta cyclotis, the forest elephant. The study, however, did not include the populations of West Africa, where the taxonomic affinities of elephants have been much debated. We examined mitochondrial cytochrome b control region sequences and four microsatellite loci to investigate the genetic differences between the forest and savannah elephants of West and Central Africa. We then combined our data with published control region sequences from across Africa to examine patterns at the continental level. Our analysis reveals several deeply divergent lineages that do not correspond with the currently recognized taxonomy: (i) the forest elephants of Central Africa; the forest and savannah elephants of West Africa; and (iii) the savannah elephants of eastern, southern and Central Africa. We propose that the complex phylogeographic patterns we detect in African elephants result from repeated continental-scale climatic changes over their five-to-six million year evolutionary history. Until there is consensus on the taxonomy, we suggest that the genetic and ecological distinctness of these lineages should be an important factor in conservation management planning.

Conservation biologists often deal with species that have small, fragmented populations throughout their range, making it necessary to prioritize populations for management. Genetics provides tools to assist with prioritization according to the levels and distribution of genetic diversity and evolutionary distinctiveness. Many studies have used nuclear microsatellite loci to measure genetic diversity in disparate populations and mitochondrial DNA to assess genetic distinctiveness. However, comparing metrics based on microsatellite genotypes ascertained in different laboratories is complicated by the selection of different loci with distinct nucleotide repeat motifs. This issue may be resolved by comparing metrics to a well-characterized reference population with shared microsatellite markers. The Asian elephant, Elephas maximus, is an endangered species with 50–60% of populations in India and Sri Lanka, and small, fragmented populations throughout southeast and insular Asia. We assessed range-wide genetic diversity of the Asian elephant by directly comparing allelic diversity and heterozygosity estimates from 35 populations, overcoming marker selection bias by calibrating metrics to a large population on the Nakai Plateau, Lao PDR, genotyped at 25 loci. We coupled these results with mtDNA analysis to evaluate genetic distinctiveness and identify potential conservation management units. We found the greatest diversity in the populations of southeast Asia and the greatest genetic distinctiveness among the subspecies designations, particularly Borneo and Sumatra, and other southeast Asian populations. The populations of southeast Asia, albeit small, fragmented, and at high risk of extirpation, contain valuable diversity and distinctiveness and are thus of high priority for the preservation of the Asian elephant. [ABSTRACT FROM AUTHOR]

Glacial cycles and pre-glacial drainage patterns have imbued the Interior Highlands of the United States with a rich suite of freshwater taxa and phylogroups. However, supplementation of sportfish from the Great Lakes into the waterbodies of this region, including the Missouri, Ouachita, Black, White, and Little Red River drainages, may have obscured phylogeographic patterns. Walleye (Sander vitreus), one of the most common sportfish in the eastern US, inhabit the Interior Highlands but their population genetic composition and structure in this region has received little attention. We examined the genetic composition of walleye (n = 643) in the Interior Highlands using microsatellite markers and a subsample (n = 188) with mitochondrial DNA and found significant genetic differences among walleye in our study area. Walleye from the Missouri, Ouachita, White, and Little Red drainages were most closely related to a Great Lakes reference sample, a common stocking source. However, the Black River, in the easternmost portion of the Interior Highlands, contained walleye with mitochondrial DNA that was closely related to walleye from the Eastern Highlands. The remainder of the study area drainages contained a mix of walleye groups, more closely related to Great Lakes rather than Highlands walleye but not definitively the product of stocking. Though managers have relied on mitochondrial markers for stock identification in the past, we recommend that walleye in regions receiving little research attention be analyzed with nuclear markers to better understand and preserve genetic diversity and that managers stock with local walleye only within the drainages that we identified as genetically distinct: Black, Missouri, and White/Little Red/Ouachita. [ABSTRACT FROM AUTHOR]

Analysis of multilocus genotypes is fundamental to conservation genetics, allowing inference ranging from population delineation to identification of dispersers; additionally, the natal range of a sample of unknown origin may be assigned. We investigated the accuracy and precision of two methods (spatial smoothing and principal components regression) for natal assignment with five datasets that varied in marker type (microsatellites or SNPs), number of loci, and number of training samples. Accuracy varied between datasets where the median difference between true and estimated geographic locations ranged from 192 to 902 km. A dataset using 1000 SNP loci and the spatial smoothing method was both the most accurate and precise. We observed that natal inference from SNPs was more accurate and precise than when estimated using microsatellites, and that large numbers of SNP loci could overcome having few samples in the training dataset. Our results suggest cautious interpretation of natal assignments, as 52% or fewer test samples were assigned to the correct management jurisdiction. In addition, samples from continuous habitat had less accurate assignments than samples from isolated areas whether due to landscape barriers or anthropogenic fragmentation. The use of natal inference as a tool for management agencies may work well at the regional level, given sufficient input data; however, we clearly observed limits on the spatial scale of inference and consequently on the effectiveness of genotypes as a sole source for natal inference. [ABSTRACT FROM AUTHOR]

Although black bears (Ursus americanus) were believed to be extirpated from the Interior Highlands of North America by the early 1900s, populations have recently recovered, aided in part by reintroductions in Arkansas. Today black bears can be found in the Ozark and Ouachita National Forests of northern and western Arkansas, the White River National Wildlife Refuge in eastern Arkansas, and the Ozark region of southern Missouri. Previous genetic studies have investigated the effects of translocating black bears from Minnesota and Manitoba, Canada, into the Ozark and Ouachita National Forests between 1958 and 1968, with differing results. We used nuclear microsatellite loci to infer the genetic structure of black bears across the Interior Highlands and to investigate the sources of bears found today in southern Missouri. Our results suggest that the Ozark population was strongly influenced by the reintroductions, whereas the Ouachita population was influenced to a lesser degree. Although the majority of bears in the Ozark region of Arkansas and Missouri represent a single genetic unit, bears in Webster County, Missouri, may represent a remnant of the historical population of the region. Our results confirm that the bear population in the White River National Wildlife Refuge is strongly differentiated genetically from other Arkansas populations and support previous reports that the Ouachita bear population may have resulted from an admixture of a remnant population and reintroduced bears. [ABSTRACT FROM AUTHOR]

We investigated the genetic metapopulation structure of elephants across the trans Rift Valley region of Kenya and Tanzania, one of the remaining strongholds for savannah elephants (Loxodonata africana) in East Africa, using microsatellite and mitochondrial DNA (mtDNA) markers. We then examined this population structure to determine the source population for a recent colonization event of savannah elephants on community-owned land within the trans rift valley region. Four of the five sampled populations showed significant genetic differentiation (p,0.05) as measured with both mtDNA haplotypes and microsatellites. Only the samples from the adjacent Maasai Mara and Serengeti ecosystems showed no significant differentiation. A phylogenetic neighbour-joining tree constructed from mtDNA haplotypes detected four clades. Clade four corresponds to the F clade of previous mtDNA studies that reported to have originated in forest elephants (Loxodonta cyclotis) but to also be present in some savannah elephant populations. The split between clade four and the other three clades corresponded strongly to the geographic distribution of mtDNA haplotypes across the rift valley in the study area. Clade four was the dominant clade detected on the west side of the rift valley with rare occurrences on the east side. Finally, the strong patterns of population differentiation clearly indicated that the recent colonists to the community-owned land in Kenya came from the west side of the rift valley. Our results indicate strong female philopatry within the isolated populations of the trans rift valley region, with gene flow primarily mediated via male movements. The recent colonization event from Maasai Mara or Serengeti suggests there is hope for maintaining connectivity and population viability outside formal protected areas in the region. [ABSTRACT FROM AUTHOR]

We screened 74 published microsatellite primers in Ambystoma annulatum, a species for which no microsatellite markers have been developed. Overall, we had a moderate success rate, identifying 11 polymorphic microsatellites previously developed in four different species of Ambystoma. We also conducted a review of the literature, collecting all published cross-species applications of microsatellite markers within the genus Ambystoma. From this, we identified 20 loci that have amplified in three or more species. Our synthesis of microsatellite use within the genus Ambystoma should prove valuable to future molecular research, especially in species without developed markers and for studies in species that may already have microsatellites, but are being conducted far from the region where individuals were collected for original development of species-specific loci. [ABSTRACT FROM AUTHOR]

Understanding factors that determine heterogeneity in levels of parasitism across individuals is a major challenge in disease ecology. It is known that genetic makeup plays an important role in infection likelihood, but the mechanism remains unclear as does its relative importance when compared to other factors. We analyzed relationships between genetic diversity and macroparasites in outbred, free-ranging populations of raccoons (Procyon lotor). We measured heterozygosity at 14 microsatellite loci and modeled the effects of both multi-locus and single-locus heterozygosity on parasitism using an information theoretic approach and including non-genetic factors that are known to influence the likelihood of parasitism. The association of genetic diversity and parasitism, as well as the relative importance of genetic diversity, differed by parasitic group. Endoparasite species richness was better predicted by a model that included genetic diversity, with the more heterozygous hosts harboring fewer endoparasite species. Genetic diversity was also important in predicting abundance of replete ticks (Dermacentor variabilis). This association fit a curvilinear trend, with hosts that had either high or low levels of heterozygosity harboring fewer parasites than those with intermediate levels. In contrast, genetic diversity was not important in predicting abundance of non-replete ticks and lice (Trichodectes octomaculatus). No strong single-locus effects were observed for either endoparasites or replete ticks. Our results suggest that in outbred populations multi-locus diversity might be important for coping with parasitism. The differences in the relationships between heterozygosity and parasitism for the different parasites suggest that the role of genetic diversity varies with parasite-mediated selective pressures. [ABSTRACT FROM AUTHOR]

Across much of North America, river otter (Lontra canadensis) populations were extirpated or greatly reduced by the early 20th century. More recently, reintroductions have resulted in restored populations and the recommencement of managed trapping. Perhaps the best example of these river otter reintroductions occurred in Missouri, regarded as one of the most successful carnivore recovery programs in history. However, abundance estimates for river otter populations are difficult to obtain and often contentious when used to underpin management activities. We assessed the value of latrine site monitoring as a mechanism for quantifying river otter abundance. Analyses of fecal DNA to identify individual animals may result in an improved population estimate and have been used for a variety of mammal species. We optimized laboratory protocols, redesigned existing microsatellite primers, and calculated genotyping error rates to enhance genotyping success for a large quantity of river otter scat samples. We also developed a method for molecular sexing. We then extracted DNA from 1,421 scat samples and anal sac secretions (anal jelly) collected during latrine site counts along 22-34-km stretches representing 8-77% of 8 rivers in southern Missouri in 2009. Error rates were low for the redesigned microsatellites. We obtained genotypes at 7-10 microsatellite loci for 24% of samples, observing highest success for anal jelly samples (71%) and lowest for fresh samples (collected within 1 day of defecation). We identified 63 otters (41 M, 22 F) in the 8 rivers, ranging from 2 to 14 otters per river. Analyses using program CAPWIRE resulted in population estimates similar to the minimum genotyping estimate. Density estimates averaged 0.24?otters/km. We used linear regression to develop and contrast models predicting population size based on latrine site and scat count indices, which are easily collected in the field. Population size was best predicted by a combination of scats per latrine and latrines per kilometer. Our results provide methodological approaches to guide wildlife managers seeking to initiate similar river otter fecal genotyping studies, as well as to estimate and monitor river otter population sizes. [ABSTRACT FROM AUTHOR]

The loggerhead shrike ( Lanius ludovicianus), a songbird that hunts like a small raptor, maintains breeding populations on seven of the eight California Channel Islands. One of the two subspecies, L. l. anthonyi, was described as having breeding populations on six of the islands while a second subspecies, L. l. mearnsi, was described as being endemic to San Clemente Island. Previous genetic studies have demonstrated that the San Clemente Island loggerhead shrike is well differentiated genetically from both L. l. anthonyi and mainland populations, despite the fact that birds from outside the population are regular visitors to the island. Those studies, however, did not include a comparison between San Clemente Island shrikes and the breeding population on Santa Catalina Island, the closest island to San Clemente. Here we use mitochondrial control region sequences and nuclear microsatellites to investigate the population structure of loggerhead shrikes in the Channel Islands. We confirm the genetic distinctiveness of the San Clemente Island loggerhead shrike and, using Bayesian clustering analysis, demonstrate the presence and infer the source of the nonbreeding visitors. Our results indicate that Channel Island loggerhead shrikes comprise three distinct genetic clusters that inhabit: (i) San Clemente Island, (ii) Santa Catalina Island and (iii) the Northern Channel Islands and nearby mainland; they do not support a recent suggestion that all Channel Island loggerhead shrikes should be managed as a single entity. [ABSTRACT FROM AUTHOR]

The hellbender is the only North American member of the aquatic salamander family Cryptobranchidae and is a species of conservation concern across its range. We developed eight polymorphic microsatellite loci for hellbenders using a magnetic bead enrichment protocol and a PCR-based detection technique. Allelic diversity averaged 4.0 (±1.8 SD) per locus and heterozygosity averaged 0.56 (±0.30 SD). The hellbender is rare and difficult to study due to its cryptic life history. These loci will provide a valuable resource for population studies, which could inform future conservation and management decisions. [ABSTRACT FROM AUTHOR]

We report the isolation and characterization of five polymorphic microsatellite loci in the Hawaii amakihi, Hemignathus virens, one of the most common native Hawaiian forest birds. These loci exhibit high levels of allelic diversity and heterozygosity in the three honeycreeper species we screened, and promise to be useful in our investigation of differences between honeycreeper populations at different elevations. [ABSTRACT FROM AUTHOR]

Bush dogs ( Speothos venaticus) are small, threatened canids from Central and South America. The ability to conduct population-level studies using noninvasive genetic samples would provide important information on this poorly understood species. We characterized eight dinucleotide microsatellite loci using samples from 15 captive bush dogs. Allelic diversity ranged from 2 to 5 alleles per locus, and observed heterozygosity ranged from 0.267 to 0.933. All loci were in Hardy–Weinberg equilibrium and no evidence of genotypic linkage disequilibrium was found. We determined that DBX6 and DBY7, two canid-specific molecular sexing primers, accurately indicate the sex of individual bush dog samples. [ABSTRACT FROM AUTHOR]