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Anna E. Savage

Publications and source records attributed to Anna E. Savage.

6 recordsLinked to original sources

Tracing invasion routes of Cuban treefrogs into Louisiana using mitochondrial DNA

Understanding the origin and spread of invasive species is critical for predicting when and where new introductions will establish, and impact native species. However, due to the complexity of contributing factors such as multiple introductions, dispersal method, genetic admixture in founding populations, and variable propagule pressure, genetic patterns observed in invasive species may not always conform to a single theoretical expectation. Cuban treefrogs ( Osteopilus septentrionalis ) are invasive in peninsular Florida and sporadically in the Florida panhandle. Though O. septentrionalis has been occasionally reported in Louisiana since the 1990s, established populations were not present until the discovery of a breeding population in New Orleans in 2017. In this study we investigated the source of this novel population using existing and newly generated cytochrome B (cyt-b) mitochondrial gene sequences from the native and invasive range of O. septentrionalis . We recovered a total of 14 cyt-b haplotypes, nine novel and five previously published. Within the 95 Louisiana invasion samples, we recovered seven haplotypes including five novel haplotypes. The haplotypes most common in Louisiana were shared exclusively with west and east Florida localities in central Florida, indicating a possible source population. The presence of haplotypes private to the Louisiana locality suggests other unsampled localities may also be contributing to the Louisiana settlement. Metrics of genetic diversity across native and invasive localities did not significantly differ. Furthermore, the Louisiana samples had higher genetic diversity than any single location sampled within Florida. Thus, genetic diversity and our haplotype connectivity suggest the Louisiana population is derived from multiple introductions from Florida. Our study highlights how demographic and genetic analyses can be utilized to understand the source and future expansion potential of invasive populations.

Florida, Louisiana

Widespread Ranavirus and Perkinsea infections in Cuban treefrogs (Osteopilus septentrionalis) invading New Orleans, USA

Invasive species can negatively impact ecosystems in numerous ways, including vectoring pathogenic organisms. In amphibians, a lineage globally threatened by multiple pathogens, this spread of disease via invasive species could contribute to declines in native populations. The Cuban Treefrog (Osteopilus septentrionalis) is invasive in the southeastern USA. To assess whether O. septentrionalis is a potential reservoir host for the pathogens Batrachochytrium dendrobatidis (Bd; Amphibian Chytrid Fungus), Ranavirus (Rv), and Perkinsea (Pr), we sampled 82 individuals from a recently invaded site in New Orleans, Louisiana, USA. We used quantitative PCR to assess prevalence and intensity of Bd, Rv, and Pr in mouthparts and tail clips from 22 larvae and in toe clips from 60 metamorphosed frogs. We compared infection prevalence and intensity across host characteristics, including Fulton’s Body Condition, sex, and life stage. None of the individuals were infected with Bd, 72% were infected with Rv, and 44% were infected with Pr. Twenty-three individuals (28%) were co-infected with Rv and Pr, but co-infection did not significantly predict the prevalence or intensity of either Rv or Pr. Although we did not observe any disease signs, Pr infections were significantly associated with lower body condition, suggesting sub-lethal fitness costs. Our study establishes that invasive O. septentrionalis in New Orleans are infected with two pathogens of global concern for amphibians. Understanding host-pathogen dynamics in O. septentrionalis in Louisiana is a critical step towards understanding how this invasive species could threaten amphibian biodiversity in the region by transmitting infectious pathogens.

Louisiana

Functional variation at an expressed MHC class IIß locus associates with Ranavirus infection intensity in larval anuran populations

Infectious diseases are causing catastrophic losses to biodiversity globally. Iridoviruses in the genus Ranavirus are among the leading causes of amphibian disease-related mortality. Polymorphisms in major histocompatibility complex (MHC) genes are significantly associated with variation in amphibian susceptibility to pathogens. MHC genes encode diverse cell-surface molecules that can recognize and bind to a wide array of pathogen peptides, and are divided into two classes. While MHC class I genes are the classic mediators of viral acquired immunity, larval amphibians do not express them. However, MHC class II gene diversity may be an important predictor of Ranavirus susceptibility in larval amphibians, the life stage most susceptible to Ranavirus. We surveyed natural populations of larval wood frogs (Lithobates sylvaticus), which are highly susceptible to Ranavirus, across 17 ponds and two years in Maryland, USA. We sequenced the peptide-binding region of an expressed MHC class IIß locus and assessed allelic and genetic diversity. We converted alleles to functional supertypes and determined if physiochemical properties of peptide-binding regions influenced host responses to Ranavirus. Among 334 sampled individuals, 26% were infected with Ranavirus, and among infected individuals the average intensity was 7.12 x 107 virus copies. We recovered 20 unique MHC class IIß alleles that fell into two deeply diverged clades and seven functional supertypes. Variation in MHC supertypes were associated with Ranavirus infection intensity, but not prevalence. MHC supertype heterozygotes and individuals with the MHC supertype genotype ST1/ST7 had significantly lower Ranavirus infection intensity compared to homozygotes and all other genotypes. We conclude that MHC class IIß functional genetic variation is an important component of Ranavirus susceptibility. Identifying immune system gene signatures linked to variation in disease susceptibility can inform mitigation strategies for combatting global amphibian declines.

Immunogenetics

Evolutionary dynamics of an expressed MHC class IIβ locus in the Ranidae (Anura) uncovered by genome walking and high-throughput amplicon sequencing

The Major Histocompatibility Complex (MHC) is a genomic region encoding immune loci that are important and frequently used markers in studies of adaptive genetic variation and disease resistance. Given the primary role of infectious diseases in contributing to global amphibian declines, we characterized the hypervariable exon 2 and flanking introns of the MHC Class IIβ chain for 17 species of frogs in the Ranidae, a speciose and cosmopolitan family facing widespread pathogen infections and declines. We find high levels of genetic variation concentrated in the Peptide Binding Region (PBR) of the exon. Ten codons are under positive selection, nine of which are located in the mammal-defined PBR. We hypothesize that the tenth codon (residue 21) is an amphibian-specific PBR site that may be important in disease resistance. Trans-species and trans-generic polymorphisms are evident from exon-based genealogies, and co-phylogenetic analyses between intron, exon and mitochondrial based reconstructions reveal incongruent topologies, likely due to different locus histories. We developed two sets of barcoded adapters that reliably amplify a single and likely functional locus in all screened species using both 454 and Illumina based sequencing methods. These primers provide a resource for multiplexing and directly sequencing hundreds of samples in a single sequencing run, avoiding the labour and chimeric sequences associated with cloning, and enabling MHC population genetic analyses. Although the primers are currently limited to the 17 species we tested, these sequences and protocols provide a useful genetic resource and can serve as a starting point for future disease, adaptation and conservation studies across a range of anuran taxa.

Developmental and Comparative Immunology

Selection, trans-species polymorphism, and locus identification of major histocompatibility complex class IIβ alleles of New World ranid frogs

Genes encoded by the major histocompatibility complex (MHC) play key roles in the vertebrate immune system. However, our understanding of the evolutionary processes and underlying genetic mechanisms shaping these genes is limited in many taxa, including amphibians, a group currently impacted by emerging infectious diseases. To further elucidate the evolution of the MHC in frogs (anurans) and develop tools for population genetics, we surveyed allelic diversity of the MHC class II ??1 domain in both genomic and complementary DNA of seven New World species in the genus Rana (Lithobates). To assign locus affiliation to our alleles, we used a "gene walking" technique to obtain intron 2 sequences that flanked MHC class II?? exon 2. Two distinct intron sequences were recovered, suggesting the presence of at least two class II?? loci in Rana. We designed a primer pair that successfully amplified an orthologous locus from all seven Rana species. In total, we recovered 13 alleles and documented trans-species polymorphism for four of the alleles. We also found quantitative evidence of selection acting on amino acid residues that are putatively involved in peptide binding and structural stability of the ??1 domain of anurans. Our results indicated that primer mismatch can result in polymerase chain reaction (PCR) bias, which influences the number of alleles that are recovered. Using a single locus may minimize PCR bias caused by primer mismatch, and the gene walking technique was an effective approach for generating single-copy orthologous markers necessary for future studies of MHC allelic variation in natural amphibian populations. ?? 2010 Springer-Verlag.

Immunogenetics

Toward immunogenetic studies of amphibian chytridiomycosis: Linking innate and acquired immunity

Recent declines in amphibian diversity and abundance have contributed significantly to the global loss of biodiversity. The fungal disease chytridiomycosis is widely considered to be a primary cause of these declines, yet the critical question of why amphibian species differ in susceptibility remains unanswered. Considerable evidence links environmental conditions and interspecific variability of the innate immune system to differential infection responses, but other sources of individual, population, or species-typical variation may also be important. In this article we review the preliminary evidence supporting a role for acquired immune defenses against chytridiomycosis, and advocate for targeted investigation of genes controlling acquired responses, as well as those that functionally bridge the innate and acquired immune systems. Immunogenetic data promise to answer key questions about chytridiomycosis susceptibility and host-pathogen coevolution, and will draw much needed attention to the importance of considering evolutionary processes in amphibian conservation management and practice. ?? 2009 by American Institute of Biological Sciences.

BioScience