Jennifer L. Bollmer

Disease ecology in the Galapagos Hawk (Buteo galapagoensis): host genetic diversity, parasite load and natural antibodies

An increased susceptibility to disease is one hypothesis explaining how inbreeding hastens extinction in island endemics and threatened species. Experimental studies show that disease resistance declines as inbreeding increases, but data from in situ wildlife systems are scarce. Genetic diversity increases with island size across the entire range of an extremely inbred Galápagos endemic bird, providing the context for a natural experiment examining the effects of inbreeding on disease susceptibility. Extremely inbred populations of Galápagos hawks had higher parasite abundances than relatively outbred populations. We found a significant island effect on constitutively produced natural antibody (NAb) levels and inbred populations generally harboured lower average and less variable NAb levels than relatively outbred populations. Furthermore, NAb levels explained abundance of amblyceran lice, which encounter the host immune system. This is the first study linking inbreeding, innate immunity and parasite load in an endemic, in situ wildlife population and provides a clear framework for assessment of disease risk in a Galápagos endemic.

Rapid loss of MHC class II variation in a bottlenecked population is explained by drift and loss of copy number variation.

Population bottlenecks may reduce genetic variation and potentially increase the risk of extinction. Here, we present the first study to use historic samples to analyse loss of variation at the major histocompatibility complex (MHC), which plays a central role in vertebrate disease resistance. Balancing selection acts on the MHC and could moderate the loss of variation expected from drift; however, in a Wisconsin population of greater prairie‐chickens (Tympanuchus cupido), the number of MHC class II B alleles per individual declined by 44% following a population bottleneck, compared to a loss of only 8% at microsatellites. Simulations indicate that drift likely reduced MHC variation at the population level, as well as within individuals by reducing the number of gene copies per individual or by fixing the same alleles across multiple loci. These multiple effects of genetic drift on MHC variation could have important implications for immunity and fitness.

Extensive MHC Class II B Gene Duplication in a Passerine, the Common Yellowthroat (Geothlypis trichas)

The major histocompatibility complex (MHC) is characterized by a birth and death model of evolution involving gene duplication, diversification, loss of function, and deletion. As a result, gene number varies across taxa. Birds have between one and 7 confirmed MHC class II B genes, and the greatest diversity appears to occur in passerines. We used multiple primer sets on both genomic DNA (gDNA) and complementary DNA (cDNA) to characterize the range of class II B genes present in a passerine, the common yellowthroat (Geothlypis trichas). We confirmed 39 exon 2 sequences from gDNA in a single individual, indicating the presence of at least 20 class II B loci. From a second individual, we recovered 16 cDNA sequences belonging to at least 8 transcribed loci. Phylogenetic analysis showed that common yellowthroat sequences fell into subgroups consisting of classical loci, as well as at least 4 different clusters of sequences with reduced sequence variability that may represent pseudogenes or nonclassical loci. Data from 2 additional common yellowthroats demonstrated high interindividual variability. Our results reveal that some passerines possess an extraordinary diversity of MHC gene duplications, including both classical and nonclassical loci.

Differentiation with drift: a spatio-temporal genetic analysis of Galápagos mockingbird populations (Mimus spp.)

Small and isolated island populations provide ideal systems to study the effects of limited population size, genetic drift and gene flow on genetic diversity. We assessed genetic diversity within and differentiation among 19 mockingbird populations on 15 Galápagos islands, covering all four endemic species, using 16 microsatellite loci. We tested for signs of drift and gene flow, and used historic specimens to assess genetic change over the last century and to estimate effective population sizes. Within-population genetic diversity and effective population sizes varied substantially among island populations and correlated strongly with island size, suggesting that island size serves as a good predictor for effective population size. Genetic differentiation among populations was pronounced and increased with geographical distance. A century of genetic drift did not change genetic diversity on an archipelago-wide scale, but genetic drift led to loss of genetic diversity in small populations, especially in one of the two remaining populations of the endangered Floreana mockingbird. Unlike in other Galápagos bird species such as the Darwins finches, gene flow among mockingbird populations was low. The clear pattern of genetically distinct populations reflects the effects of genetic drift and suggests that Galápagos mockingbirds are evolving in relative isolation.

MHC VARIATION IS RELATED TO A SEXUALLY SELECTED ORNAMENT, SURVIVAL, AND PARASITE RESISTANCE IN COMMON YELLOWTHROATS

Hamilton and Zuk proposed that females choose mates based on ornaments whose expression is dependent on their genetically based resistance to parasites. The major histocompatibility complex (MHC) plays an important role in pathogen recognition and is a good candidate for testing the relationships between immune genes and both ornament expression and parasite resistance. We tested the hypothesis that female common yellowthroats prefer to mate with more ornamented males, because it is a signal of their MHC‐based resistance to parasites and likelihood of survival. In this species, females prefer males that have larger black facial masks as extrapair mates. Using pyrosequencing, we found that mask size was positively related to the number of different MHC class II alleles, as predicted if greater variation at the MHC allows for the recognition of a greater variety of pathogens. Furthermore, males with more MHC class II alleles had greater apparent survival, and resistance to malaria infection was associated with the presence of a particular MHC class II allele. Thus, extrapair mating may provide female warblers with immunity genes that are related to parasite resistance, survival, and the expression of a male ornament, consistent with good genes models of sexual selection.

Drift and selection influence geographic variation at immune loci of prairie‐chickens

Previous studies of immunity in wild populations have focused primarily on genes of the major histocompatibility complex (MHC); however, studies of model species have identified additional immune‐related genes that also affect fitness. In this study, we sequenced five non‐MHC immune genes in six greater prairie‐chicken (Tympanuchus cupido) populations that have experienced varying degrees of genetic drift as a consequence of population bottlenecks and fragmentation. We compared patterns of geographic variation at the immune genes with six neutral microsatellite markers to investigate the relative effects of selection and genetic drift. Global FST outlier tests identified positive selection on just one of five immune genes (IAP‐1) in one population. In contrast, at other immune genes, standardized G′ST values were lower than those at microsatellites for a majority of pairwise population comparisons, consistent with balancing selection or with species‐wide positive or purifying selection resulting in similar haplotype frequencies across populations. The effects of genetic drift were also evident as summary statistics (e.g., Tajima’s D) did not differ from neutrality for the majority of cases, and immune gene diversity (number of haplotypes per gene) was correlated positively with population size. In summary, we found that both genetic drift and selection shaped variation at the five immune genes, and the strength and type of selection varied among genes. Our results caution that neutral forces, such as drift, can make it difficult to detect current selection on genes.

VARIATION IN MORPHOLOGY AND MATING SYSTEM AMONG ISLAND POPULATIONS OF GALÁPAGOS HAWKS

Abstract Interspecific variation in sexual size dimorphism has commonly been attributed to variation in social mating system, with dimorphism increasing as intrasexual competition for mates increases. In birds, overall body size has also been found to correlate positively with size dimorphism. In this study, we describe variation in morphology and mating system across six populations of the endemic Galápagos Hawk (Buteo galapagoensis). Galápagos Hawks exhibit cooperative polyandry, a mating system in which long-term social groups contain a single female and multiple males. Comparisons among islands revealed significant differences in overall body size for both adults and immatures. Populations ranged from completely monogamous to completely polyandrous, with varying mean group sizes. Data did not support our prediction that sexual size dimorphism would increase with the degree of polyandry (number of males per group) or with body size; there was no correlation between mating system and sexual dimorphism. We did find a significant negative relationship between degree of polyandry and body size among islands, opposite of the pattern predicted. Variación en Morfología y Sistema de Apareamiento entre Poblaciones de Buteo galapagoensis Resumen. Variación interespecífica en dimorfismo sexual ha sido atribuída comúnmente a variaciones del sistema social de apareamiento, de tal manera que el dimorfismo aumenta conforme aumenta la competencia intrasexual por parejas reproductivas. También se ha encontrado que el tamaño corporal se correlaciona positivamente con el dimorfismo. En este estudio describimos la variación morfológica y el grado de poliandría de seis poblaciones de Buteo galapagoensis, una especie que exhibe un sistema de apareamiento denominado poliandría cooperativa. En este sistema los grupos de individuos reproductivos incluyen una sola hembra y múltiples machos. Se comprobó que existen diferencias significativas en el tamaño del cuerpo de adultos y juveniles entre islas. Las poblaciones muestreadas variaron entre monógamas y completamente poliándricas, y el tamaño promedio de los grupos fue variable. Los datos no apoyaron las predicciones establecidas inicialmente pues el grado de dimorfismo sexual no aumentó con el nivel de poliandría (número de machos por grupo) ni con el tamaño corporal, ni hubo una correlación entre el sistema de apareamiento y el dimorfismo sexual. La relación entre el tamaño corporal y el sistema de apareamiento fue contraria a la que se predijo: hubo una correlación negativa significativa entre el grado de poliandría y el tamaño corporal entre islas.

Social and extra-pair mating in relation to major histocompatibility complex variation in common yellowthroats

Females are thought to gain better-quality genes for their offspring by mating with particular males. Genes of the major histocompatibility complex (MHC) play a critical role in adaptive immunity, and several studies have examined female mate choice in relation to MHC variation. In common yellowthroats, females prefer males that have larger black facial masks, an ornament associated with MHC variation, immune function and condition. Here we also tested whether mating patterns are directly correlated with MHC diversity or similarity. Using pyrosequencing, we found that the presence of extra-pair young in the brood was not related to male MHC diversity or similarity between the female and her within-pair mate. Furthermore, extra-pair sires did not differ in overall diversity from males they cuckolded, or in their similarity to the female. MHC diversity is extremely high in this species, and it may limit the ability of females to assess MHC variation in males. Thus, mating may be based on ornaments, such as mask size, which are better indicators of overall male health and genetic quality.

Reduced MHC and neutral variation in the Galapagos hawk, an island endemic

BackgroundGenes at the major histocompatibility complex (MHC) are known for high levels of polymorphism maintained by balancing selection. In small or bottlenecked populations, however, genetic drift may be strong enough to overwhelm the effect of balancing selection, resulting in reduced MHC variability. In this study we investigated MHC evolution in two recently diverged bird species: the endemic Galápagos hawk (Buteo galapagoensis), which occurs in small, isolated island populations, and its widespread mainland relative, the Swainsons hawk (B. swainsoni).ResultsWe amplified at least two MHC class II B gene copies in each species. We recovered only three different sequences from 32 Galápagos hawks, while we amplified 20 unique sequences in 20 Swainsons hawks. Most of the sequences clustered into two groups in a phylogenetic network, with one group likely representing pseudogenes or nonclassical loci. Neutral genetic diversity at 17 microsatellite loci was also reduced in the Galápagos hawk compared to the Swainsons hawk.ConclusionsThe corresponding loss in neutral diversity suggests that the reduced variability present at Galápagos hawk MHC class II B genes compared to the Swainsons hawk is primarily due to a founder event followed by ongoing genetic drift in small populations. However, purifying selection could also explain the low number of MHC alleles present. This lack of variation at genes involved in the adaptive immune response could be cause for concern should novel diseases reach the archipelago.

Greater prairie chickens have a compact MHC-B with a single class IA locus

The major histocompatibility complex (MHC) plays a central role in innate and adaptive immunity, but relatively little is known about the evolution of the number and arrangement of MHC genes in birds. Insights into the evolution of the MHC in birds can be gained by comparing the genetic architecture of the MHC between closely related species. We used a fosmid DNA library to sequence a 60.9-kb region of the MHC of the greater prairie chicken (Tympanuchus cupido), one of five species of Galliformes with a physically mapped MHC. Greater prairie chickens have the smallest core MHC yet observed in any bird species, and major changes are observed in the number and arrangement of MHC loci. In particular, the greater prairie chicken differs from other Galliformes in the deletion of an important class I antigen binding gene. Analysis of the remaining class IA gene in a population of greater prairie chickens in Wisconsin, USA revealed little evidence for selection at the region responsible for antigen binding.