Severe Bottleneck Research Articles

Contrasting demographic histories in a pair of allopatric, sibling species of fish (Arripidae) from environments with contrasting glacial histories

Although Pleistocene glacial cycles are regularly used to explain many aspects of the demographic history of coastal marine species, the present study is one of few to test a priori predictions about these effects. Nucleotide sequence variation in the cytochrome b gene of the mtDNA and allele frequency variation at intron loci in the nDNA were compared between two species of Arripis (Australian Salmon) that are closely related and biologically similar other than for their allopatric distributions. The results suggest that A. truttaceus, which resides to the west of the Bass Strait, exhibits lower levels of genetic diversity and experienced a severe population bottleneck during the LGM followed by an expansion commencing some 17,000 years ago. In contrast, the population of A. trutta, which resides to the east of the Bass Strait, appears to have been largely unaffected by the LGM and has been expanding over the past 100,000 years or more. These results are consistent with a priori predictions, based on paleo-oceanographic data, that the demographic history of A. truttaceus has been more strongly affected by glacial periods by virtue of its distribution to the west of Bass Strait. Data on two other congeners are also presented to provide context for the results for A. trutta and A. truttaceus.

GPU-accelerated ray-tracing for real-time treatment planning

Dose calculation methods in radiotherapy treatment planning require the radiological depth information of the voxels that represent the patient volume to correct for tissue inhomogeneities. This information is acquired by time consuming ray-tracing-based calculations. For treatment planning scenarios with changing geometries and real-time constraints this is a severe bottleneck. We implemented an algorithm for the graphics processing unit (GPU) which implements a ray-matrix approach to reduce the number of rays to trace. Furthermore, we investigated the impact of different strategies of accessing memory in kernel implementations as well as strategies for rapid data transfers between main memory and memory of the graphics device. Our study included the overlapping of computations and memory transfers to reduce the overall runtime using Hyper-Q. We tested our approach on a prostate case (9 beams, coplanar). The measured execution times for a complete ray-tracing range from 28 msec for the computations on the GPU to 99 msec when considering data transfers to and from the graphics device. Our GPU-based algorithm performed the ray-tracing in real-time. The strategies efficiently reduce the time consumption of memory accesses and data transfer overhead. The achieved runtimes demonstrate the viability of this approach and allow improved real-time performance for dose calculation methods in clinical routine.

Maintenance of genetic diversity in an introduced island population of guanacos after seven decades and two severe demographic bottlenecks: implications for camelid conservation.

Fifteen guanacos were introduced to Staats Island in the Falklands/Malvinas archipelago from Patagonia in the 1930s. Twenty five years later, the population was culled from 300 to 10–20 individuals, but quickly rebounded to a population of almost 400 animals that today retain the genetic signature of the founding event and later bottleneck. The goals of this study were to (i) make a genetic assessment of this island population through comparisons with mainland populations and simulations, and (ii) assess the likely source-population of the introduced guanacos. Genetic variation was estimated from 513 bp of mitochondrial DNA sequence and 15 microsatellite loci among 154 guanacos collected from eight localities, including the adjacent mainland and the islands of Tierra del Fuego and Staats Island. Of the 23 haplotypes observed among our samples, the Staats Island population only contained three haplotypes, all of which were shared with the coastal Monte Leon population in southern Patagonia. Mitochondrial DNA and microsatellite variations on Staats Island were comparable to most mainland populations and greater than those observed on Tierra del Fuego. Patterns of genetic structure suggest that the Staats Island guanaco population was founded with animals from southern Patagonia (as opposed to northern Patagonia or Tierra del Fuego), but that effective reductions in population size lasted only a few generations and that surviving animals were a random sample of the pre-bottleneck genetic variation.

Combining genetic data to identify relatedness among founders in a genetically depauperate parrot, the Kakapo (Strigops habroptilus)

Small populations that have experienced a bottleneck often suffer from low genetic diversity and inbreeding, the effects of which may further threaten persistence over time. The iconic Kakapo (Strigops habroptilus) is an endemic parrot of New Zealand, which was nearly driven to extinction by introduced predators and has subsequently experienced a severe bottleneck of 60 individuals. The population has since recovered to 125 individuals, but exhibits high hatching failure and other signs of inbreeding depression. Previous studies have indicated that contemporary Kakapo possess low genetic diversity, thus maintaining existing genetic diversity is high priority in Kakapo management. The future plans are to manage Kakapo using a pedigree-based system. An assumption common to most pedigrees is that founding individuals are unrelated. However, this is unlikely to be the case for Kakapo, as most founders originate from a small area of Stewart Island. Therefore the goal of this study was to examine the current Kakapo founders for potential relationships using several molecular techniques. Founders were genotyped at 25 microsatellites, which were used to generate relatedness estimates and assign relationships. 94 pedigree relationships were identified among founders comprising full sibling and half sibling pairs, as well as 110 with high pairwise relatedness values indicating likely close relationships. These relationships, along with mtDNA control region haplotypes, were combined to inform the pedigree. The improved pedigree will be used to manage the Kakapo population genetically through artificial insemination and planned matings.

Temporal genetic structure of major dengue vector Aedes aegypti from Manaus, Amazonas, Brazil

In recent years, high levels of Aedes aegypti infestation and several dengue outbreaks with fatal outcome cases have been reported in Manaus, State of Amazonas, Brazil. This situation made it important to understand the genetic structure and gene flow patterns among the populations of this vector in Manaus, vital pieces of information for their management and development of new control strategies. In this study, we used nine microsatellite loci to examine the effect of seasonality on the genetic structure and gene flow patterns in Ae. aegypti populations from four urban neighborhoods of Manaus, collected during the two main rainy and dry seasons. All loci were polymorphic in the eight samples from the two seasons, with a total of 41 alleles. The genetic structure analyses of the samples from the rainy season revealed genetic homogeneity and extensive gene flow, a result consistent with the abundance of breeding sites for this vector. However, the samples from the dry season were significantly structured, due to a reduction of Ne in two (Praça 14 de Janeiro and Cidade Nova) of the four samples analyzed, and this was the primary factor influencing structure during the dry season. Genetic bottleneck analyses suggested that the Ae. aegypti populations from Manaus are being maintained continuously throughout the year, with seasonal reduction rather than severe bottleneck or extinction, corroborating previous reports. These findings are of extremely great importance for designing new dengue control strategies in Manaus.

The number and genetic relatedness of transmitted/founder virus impact clinical outcome in vaginal R5 SHIVSF162P3N infection

BackgroundSevere genetic bottleneck occurs during HIV-1 sexual transmission whereby most infections are initiated by a single transmitted/founder (T/F) virus. Similar observations had been made in nonhuman primates exposed mucosally to SIV/SHIV. We previously reported variable clinical outcome in rhesus macaques inoculated intravaginally (ivg) with a high dose of R5 SHIVSF162P3N. Given the potential contributions of viral diversity to HIV-1 persistence and AIDS pathogenesis and recombination between retroviral genomes increases the genetic diversity, we tested the hypothesis that transmission of multiple variants contributes to heightened levels of virus replication and faster disease progression in the SHIVSF162P3N ivg-infected monkeys.ResultsWe found that the differences in viral replication and disease progression between the transiently viremic (TV; n = 2), chronically-infected (CP; n = 8) and rapid progressor (RP; n = 4) ivg-infected macaques cannot be explained by which variant in the inoculum was infecting the animal. Rather, transmission of a single variant was observed in both TV rhesus, with 1–2 T/F viruses found in the CPs and 2–4 in all four RP macaques. Moreover, the genetic relatedness of the T/F viruses in the CP monkeys with multivariant transmission was greater than that seen in the RPs. Biological characterization of a subset of T/F envelopes from chronic and rapid progressors revealed differences in their ability to mediate entry into monocyte-derived macrophages, with enhanced macrophage tropism observed in the former as compared to the latter.ConclusionOur study supports the tenet that sequence diversity of the infecting virus contributes to higher steady-state levels of HIV-1 virus replication and faster disease progression and highlights the role of macrophage tropism in HIV-1 transmission and persistence.

Optimizing Massive Distance Computations in Pattern Recognition

It is a common task in pattern recognition to evaluate the similarity of large data objects. These are often represented by high dimensional vectors. A frequently used mathematical model for evaluating their similarity is to view them as points (vectors) in a high dimensional space, and compute their distances from each other. The distance, however, can be defined in a very complicated way, it may be much more complex than the well known Euclidean distance. Therefore, the algorithmic bottleneck often becomes the number of distance computations that need to be carried out. We consider the case when we have to compute all the distances between n objects, where n is large. Without any shortcuts it takes n ( n − 1)/2 = O ( n 2 ) distance computations. In those applications where the distances are complicated, being defined by sophisticated algorithms (such as in speech and image recognition), a quadratically growing number of distance computations becomes a severe bottleneck. We prove the following general result that can help eliminating the bottleneck: for a large and general class of distances it is possible to obtain a very close approximation of each of the O(n 2 ) pairwise distances of n objects by doing only a linear number distance computations, which is optimal with respect to the order of magnitude. Moreover, the approximation factor can be made arbitrarily close to 1, making the approximation error negligible. The needed side computations to achieve this reduction can also be done in polynomial time.

Genome Sequencing Highlights the Dynamic Early History of Dogs

To identify genetic changes underlying dog domestication and reconstruct their early evolutionary history, we generated high-quality genome sequences from three gray wolves, one from each of the three putative centers of dog domestication, two basal dog lineages (Basenji and Dingo) and a golden jackal as an outgroup. Analysis of these sequences supports a demographic model in which dogs and wolves diverged through a dynamic process involving population bottlenecks in both lineages and post-divergence gene flow. In dogs, the domestication bottleneck involved at least a 16-fold reduction in population size, a much more severe bottleneck than estimated previously. A sharp bottleneck in wolves occurred soon after their divergence from dogs, implying that the pool of diversity from which dogs arose was substantially larger than represented by modern wolf populations. We narrow the plausible range for the date of initial dog domestication to an interval spanning 11–16 thousand years ago, predating the rise of agriculture. In light of this finding, we expand upon previous work regarding the increase in copy number of the amylase gene (AMY2B) in dogs, which is believed to have aided digestion of starch in agricultural refuse. We find standing variation for amylase copy number variation in wolves and little or no copy number increase in the Dingo and Husky lineages. In conjunction with the estimated timing of dog origins, these results provide additional support to archaeological finds, suggesting the earliest dogs arose alongside hunter-gathers rather than agriculturists. Regarding the geographic origin of dogs, we find that, surprisingly, none of the extant wolf lineages from putative domestication centers is more closely related to dogs, and, instead, the sampled wolves form a sister monophyletic clade. This result, in combination with dog-wolf admixture during the process of domestication, suggests that a re-evaluation of past hypotheses regarding dog origins is necessary.

Modeling and Simulation of Complex Network Attributes on Coordinating Large Multiagent System

With the expansion of distributed multiagent systems, traditional coordination strategy becomes a severe bottleneck when the system scales up to hundreds of agents. The key challenge is that in typical large multiagent systems, sparsely distributed agents can only communicate directly with very few others and the network is typically modeled as an adaptive complex network. In this paper, we present simulation testbed CoordSim built to model the coordination of network centric multiagent systems. Based on the token-based strategy, the coordination can be built as a communication decision problem that agents make decisions to target communications and pass them over to the capable agents who will potentially benefit the team most. We have theoretically analyzed that the characters of complex network make a significant difference with both random and intelligent coordination strategies, which may contribute to future multiagent algorithm design.

Phylogeography and trans‐Pacific divergence of the rocky shore gastropod Nucella lima

AbstractAimWe have used phylogeographical and multilocus coalescent population genetic methods to reconstruct the Pleistocene biogeographical history of a broadly distributed north Pacific rocky shore gastropod, Nucella lima.LocationNorthern Pacific rim, from south‐eastern Alaska to Hokkaido Island.MethodsWe gathered DNA sequence data from three loci from N. lima, whose current distribution spans the entire North Pacific rim. We used a combination of population genetic summary statistics (Tajima's D, Fu's FS and ΦST), isolation‐with‐migration divergence models, and extended Bayesian skyline plots to reconstruct the recent biogeographical history of this species.ResultsThe largest values of ΦST across all loci were always between eastern and western samples. Population divergence models indicated no gene flow and mid‐Pleistocene divergence times (> 600 ka) between eastern and western populations. Mitochondrial DNA diversity in the east was low, and coalescent‐based estimates of effective population size were significantly smaller in the east 20,000 years ago, at the end of the last glacial period.Main conclusionsThe results are consistent with a hypothesis in which north Pacific populations were separated into eastern and western refugia by 317 ka with no gene flow since the split. Eastern populations probably underwent a severe bottleneck in population size during the last glacial period. The contrasting demographic histories of eastern and western populations are consistent with the general palaeobiogeographical pattern of greater climate‐related extinction of marine taxa in the eastern Pacific.

SNP genotyping reveals genetic diversity between cultivated landraces and contemporary varieties of tomato

BackgroundThe tomato (Solanum lycopersium L.) is the most widely grown vegetable in the world. It was domesticated in Latin America and Italy and Spain are considered secondary centers of diversification. This food crop has experienced severe genetic bottlenecks and modern breeding activities have been characterized by trait introgression from wild species and divergence in different market classes.ResultsWith the aim to examine patterns of polymorphism, characterize population structure and identify putative loci under positive selection, we genotyped 214 tomato accessions (which include cultivated landraces, commercial varieties and wild relatives) using a custom-made Illumina SNP-panel. Most of the 175 successfully scored SNP loci were found to be polymorphic. Population structure analysis and estimates of genetic differentiation indicated that landraces constitute distinct sub-populations. Furthermore, contemporary varieties could be separated in groups (processing, fresh and cherry) that are consistent with the recent breeding aimed at market-class specialization. In addition, at the 95% confidence level, we identified 30, 34 and 37 loci under positive selection between landraces and each of the groups of commercial variety (cherry, processing and fresh market, respectively). Their number and genomic locations imply the presence of some extended regions with high genetic variation between landraces and contemporary varieties.ConclusionsOur work provides knowledge concerning the level and distribution of genetic variation within cultivated tomato landraces and increases our understanding of the genetic subdivision of contemporary varieties. The data indicate that adaptation and selection have led to a genomic signature in cultivated landraces and that the subpopulation structure of contemporary varieties is shaped by directed breeding and largely of recent origin. The genomic characterization presented here is an essential step towards a future exploitation of the available tomato genetic resources in research and breeding programs.Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-14-835) contains supplementary material, which is available to authorized users.

Invasion Pathway of the Ctenophore Mnemiopsis leidyi in the Mediterranean Sea

Gelatinous zooplankton outbreaks have increased globally owing to a number of human-mediated factors, including food web alterations and species introductions. The invasive ctenophore Mnemiopsis leidyi entered the Black Sea in the early 1980s. The invasion was followed by the Azov, Caspian, Baltic and North Seas, and, most recently, the Mediterranean Sea. Previous studies identified two distinct invasion pathways of M. leidyi from its native range in the western Atlantic Ocean to Eurasia. However, the source of newly established populations in the Mediterranean Sea remains unclear. Here we build upon our previous study and investigate sequence variation in both mitochondrial (Cytochrome c Oxidase subunit I) and nuclear (Internal Transcribed Spacer) markers in M. leidyi, encompassing five native and 11 introduced populations, including four from the Mediterranean Sea. Extant genetic diversity in Mediterranean populations (n = 8, N a = 10) preclude the occurrence of a severe genetic bottleneck or founder effects in the initial colonizing population. Our mitochondrial and nuclear marker surveys revealed two possible pathways of introduction into Mediterranean Sea. In total, 17 haplotypes and 18 alleles were recovered from all surveyed populations. Haplotype and allelic diversity of Mediterranean populations were comparable to populations from which they were likely drawn. The distribution of genetic diversity and pattern of genetic differentiation suggest initial colonization of the Mediterranean from the Black-Azov Seas (pairwise F ST = 0.001–0.028). However, some haplotypes and alleles from the Mediterranean Sea were not detected from the well-sampled Black Sea, although they were found in Gulf of Mexico populations that were also genetically similar to those in the Mediterranean Sea (pairwise F ST = 0.010–0.032), raising the possibility of multiple invasion sources. Multiple introductions from a combination of Black Sea and native region sources could be facilitated by intense local and transcontinental shipping activity, respectively.

Impact on Reindeer (Rangifer tarandus) Genetic Diversity from Two Parallel Population Bottlenecks Founded from a Common Source

Population bottlenecks and founder events reduce genetic diversity through stochastic processes associated with the sampling of alleles at the time of the bottleneck, and the recombination of alleles that are identical by descent. At the same time bottlenecks and founder events can structure populations through the stochastic distortion of allele frequencies. Here we undertake an empirical assessment of the impact of two independent bottlenecks of known size from a known source, and consider inference about evolutionary process in the context of simulations and theoretical expectations. We find a similar level of reduced variation in the parallel bottleneck events, with the greater impact on the population that began with the smaller number of females. The level of diversity remaining was consistent with model predictions, but only if re-growth of the population was essentially exponential and polygeny was minimal at the early stages. There was a high level of differentiation seen compared to the source population and between the two bottlenecked populations, reflecting the stochastic distortion of allele frequencies. We provide empirical support for the theoretical expectations that considerable diversity can remain following a severe bottleneck event, given rapid demographic recovery, and that populations founded from the same source can become quickly differentiated. These processes may be important during the evolution of population genetic structure for species affected by rapid changes in available habitat.

Origins and domestication of cultivated banana inferred from chloroplast and nuclear genes.

BackgroundCultivated bananas are large, vegetatively-propagated members of the genus Musa. More than 1,000 cultivars are grown worldwide and they are major economic and food resources in numerous developing countries. It has been suggested that cultivated bananas originated from the islands of Southeast Asia (ISEA) and have been developed through complex geodomestication pathways. However, the maternal and parental donors of most cultivars are unknown, and the pattern of nucleotide diversity in domesticated banana has not been fully resolved.Methodology/Principal FindingsWe studied the genetics of 16 cultivated and 18 wild Musa accessions using two single-copy nuclear (granule-bound starch synthase I, GBSS I, also known as Waxy, and alcohol dehydrogenase 1, Adh1) and two chloroplast (maturase K, matK, and the trnL-F gene cluster) genes. The results of phylogenetic analyses showed that all A-genome haplotypes of cultivated bananas were grouped together with those of ISEA subspecies of M. acuminata (A-genome). Similarly, the B- and S-genome haplotypes of cultivated bananas clustered with the wild species M. balbisiana (B-genome) and M. schizocarpa (S-genome), respectively. Notably, it has been shown that distinct haplotypes of each cultivar (A-genome group) were nested together to different ISEA subspecies M. acuminata. Analyses of nucleotide polymorphism in the Waxy and Adh1 genes revealed that, in comparison to the wild relatives, cultivated banana exhibited slightly lower nucleotide diversity both across all sites and specifically at silent sites. However, dramatically reduced nucleotide diversity was found at nonsynonymous sites for cultivated bananas.Conclusions/SignificanceOur study not only confirmed the origin of cultivated banana as arising from multiple intra- and inter-specific hybridization events, but also showed that cultivated banana may have not suffered a severe genetic bottleneck during the domestication process. Importantly, our findings suggested that multiple maternal origins and a reduction in nucleotide diversity at nonsynonymous sites are general attributes of cultivated bananas.

Toward unsupervised single-shot diffractive imaging of heterogeneous particles using X-ray free-electron lasers

Single shot diffraction imaging experiments via X-ray free-electron lasers can generate as many as hundreds of thousands of diffraction patterns of scattering objects. Recovering the real space contrast of a scattering object from these patterns currently requires a reconstruction process with user guidance in a number of steps, introducing severe bottlenecks in data processing. We present a series of measures that replace user guidance with algorithms that reconstruct contrasts in an unsupervised fashion. We demonstrate the feasibility of automating the reconstruction process by generating hundreds of contrasts obtained from soot particle diffraction experiments.

Demographic Divergence History of Pied Flycatcher and Collared Flycatcher Inferred from Whole-Genome Re-sequencing Data

Profound knowledge of demographic history is a prerequisite for the understanding and inference of processes involved in the evolution of population differentiation and speciation. Together with new coalescent-based methods, the recent availability of genome-wide data enables investigation of differentiation and divergence processes at unprecedented depth. We combined two powerful approaches, full Approximate Bayesian Computation analysis (ABC) and pairwise sequentially Markovian coalescent modeling (PSMC), to reconstruct the demographic history of the split between two avian speciation model species, the pied flycatcher and collared flycatcher. Using whole-genome re-sequencing data from 20 individuals, we investigated 15 demographic models including different levels and patterns of gene flow, and changes in effective population size over time. ABC provided high support for recent (mode 0.3 my, range <0.7 my) species divergence, declines in effective population size of both species since their initial divergence, and unidirectional recent gene flow from pied flycatcher into collared flycatcher. The estimated divergence time and population size changes, supported by PSMC results, suggest that the ancestral species persisted through one of the glacial periods of middle Pleistocene and then split into two large populations that first increased in size before going through severe bottlenecks and expanding into their current ranges. Secondary contact appears to have been established after the last glacial maximum. The severity of the bottlenecks at the last glacial maximum is indicated by the discrepancy between current effective population sizes (20,000–80,000) and census sizes (5–50 million birds) of the two species. The recent divergence time challenges the supposition that avian speciation is a relatively slow process with extended times for intrinsic postzygotic reproductive barriers to evolve. Our study emphasizes the importance of using genome-wide data to unravel tangled demographic histories. Moreover, it constitutes one of the first examples of the inference of divergence history from genome-wide data in non-model species.

Estimating relatedness and inbreeding using molecular markers and pedigrees: the effect of demographic history

Estimates of inbreeding and relatedness are commonly calculated using molecular markers, although the accuracy of such estimates has been questioned. As a further complication, in many situations, such estimates are required in populations with reduced genetic diversity, which is likely to affect their accuracy. We investigated the correlation between microsatellite- and pedigree-based coefficients of inbreeding and relatedness in laboratory populations of Drosophila melanogaster that had passed through bottlenecks to manipulate their genetic diversity. We also used simulations to predict expected correlations between marker- and pedigree-based estimates and to investigate the influence of linkage between loci and null alleles. Our empirical data showed lower correlations between marker- and pedigree-based estimates in our control (nonbottleneck) population than were predicted by our simulations or those found in similar studies. Correlations were weaker in bottleneck populations, confirming that extreme reductions in diversity can compromise the ability of molecular estimates to detect recent inbreeding events. However, this result was highly dependent on the strength of the bottleneck and we did not observe or predict any reduction in correlations in our population that went through a relatively severe bottleneck of N=10 for one generation. Our results are therefore encouraging, as molecular estimates appeared robust to quite severe reductions in genetic diversity. It should also be remembered that pedigree-based estimates may not capture realized identity-by-decent and that marker-based estimates may actually be more useful in certain situations.

Ancient DNA and Population Turnover in Southern Levantine Pigs- Signature of the Sea Peoples Migration?

Near Eastern wild boars possess a characteristic DNA signature. Unexpectedly, wild boars from Israel have the DNA sequences of European wild boars and domestic pigs. To understand how this anomaly evolved, we sequenced DNA from ancient and modern pigs from Israel. Pigs from Late Bronze Age (until ca. 1150 BCE) in Israel shared haplotypes of modern and ancient Near Eastern pigs. European haplotypes became dominant only during the Iron Age (ca. 900 BCE). This raises the possibility that European pigs were brought to the region by the Sea Peoples who migrated to the Levant at that time. Then, a complete genetic turnover took place, most likely because of repeated admixture between local and introduced European domestic pigs that went feral. Severe population bottlenecks likely accelerated this process. Introductions by humans have strongly affected the phylogeography of wild animals, and interpretations of phylogeography based on modern DNA alone should be taken with caution.

Unregulated hunting and genetic recovery from a severe population decline: the cautionary case of Bulgarian wolves

European wolf (Canis lupus) populations have suffered extensive decline and range contraction due to anthropogenic culling. In Bulgaria, although wolves are still recovering from a severe demographic bottleneck in the 1970s, hunting is allowed with few constraints. A recent increase in hunting pressure has raised concerns regarding long-term viability. We thus carried out a comprehensive conservation genetic analysis using microsatellite and mtDNA markers. Our results showed high heterozygosity levels (0.654, SE 0.031) and weak genetic bottleneck signals, suggesting good recovery since the 1970s decline. However, we found high levels of inbreeding (F IS = 0.113, SE 0.019) and a N e/N ratio lower than expected for an undisturbed wolf population (0.11, 95 % CI 0.08–0.29). We also found evidence for hybridisation and introgression from feral dogs (C. familiaris) in 10 out of 92 wolves (9.8 %). Our results also suggest admixture between wolves and local populations of golden jackals (C. aureus), but less extensive as compared with the admixture with dogs. We detected local population structure that may be explained by fragmentation patterns during the 1970s decline and differences in local ecological characteristics, with more extensive sampling needed to assess further population substructure. We conclude that high levels of inbreeding and hybridisation with other canid species, which likely result from unregulated hunting, may compromise long-term viability of this population despite its current high genetic diversity. The existence of population subdivision warrants an assessment of whether separate management units are needed for different subpopulations. Our study highlights conservation threats for populations with growing numbers but subject to unregulated hunting.

Deep Sequencing of Viral Genomes Provides Insight into the Evolution and Pathogenesis of Varicella Zoster Virus and Its Vaccine in Humans

Immunization with the vOka vaccine prevents varicella (chickenpox) in children and susceptible adults. The vOka vaccine strain comprises a mixture of genotypes and, despite attenuation, causes rashes in small numbers of recipients. Like wild-type virus, the vaccine establishes latency in neuronal tissue and can later reactivate to cause Herpes zoster (shingles). Using hybridization-based methodologies, we have purified and sequenced vOka directly from skin lesions. We show that alleles present in the vaccine can be recovered from the lesions and demonstrate the presence of a severe bottleneck between inoculation and lesion formation. Genotypes in any one lesion appear to be descended from one to three vaccine-genotypes with a low frequency of novel mutations. No single vOka haplotype and no novel mutations are consistently present in rashes, indicating that neither new mutations nor recombination with wild type are critical to the evolution of vOka rashes. Instead, alleles arising from attenuation (i.e., not derived from free-living virus) are present at lower frequencies in rash genotypes. We identify 11 loci at which the ancestral allele is selected for in vOka rash formation and show genotypes in rashes that have reactivated from latency cannot be distinguished from rashes occurring immediately after inoculation. We conclude that the vOka vaccine, although heterogeneous, has not evolved to form rashes through positive selection in the mode of a quasispecies, but rather alleles that were essentially neutral during the vaccine production have been selected against in the human subjects, allowing us to identify key loci for rash formation.