Patricia G. Parker

WHAT MOLECULES CAN TELL US ABOUT POPULATIONS: CHOOSING ANDUSING A MOLECULAR MARKER

The rapid development of molecular techniques offers a palette of technical approaches for population biologists interested in a wide range of questions. For example, these tools can be used to determine individual reproductive success or to measure rates of genetic divergence among populations. Which technique is most appropriate for a par- ticular question depends upon (1) the extent of genetic polymorphism required to best answer the question, (2) the analytical or statistical approaches available for the techniques application, and (3) the pragmatics of time and costs of materials. Here we evaluate the application of several major techniques (protein electrophoresis, nuclear and mitochondrial RFLPs (restriction fragment length polymorphisms), minisatellite and microsatellite VNTRs (variable number tandem repeats), RAPDs (random amplified polymorphic DNA), and DNA sequencing) to an array of questions regarding individual identification, exclusion and assignment of parentage, and various levels of population structure. In our evaluation, we briefly explain the technical components of each molecular approach and assess whether the typical outcomes expected from each approach will provide useful information as applied to each level of inquiry. For studies of population genetic structure, protein electrophoresis remains a powerful tool for most taxa, although techniques based on nucleic acids (par- ticularly DNA sequencing and mitochondrial DNA RFLPs) are useful here as well. Recently developed nucleic acid techniques (e.g., VNTRs) can often identify enough genetic vari- ability to address questions of self-identification or parentage. Some of the newest tech- niques (RAPDs and microsatellites) are potentially useful across a number of levels of inquiry, although procedures for adopting them are still developing.

Testosterone affects reproductive success by influencing extra–pair fertilizations in male dark–eyed juncos (Aves: Junco hyemalis)

Monogamous male birds typically allocate less effort to courtship and more to parental behaviour than males of polygynous species. The seasonal pattern of testosterone (T) secretion varies accordingly. Monogamous males exhibit a spring peak in plasma T followed by lower levels during the parental phase, while males of polygynous species continue to court females and maintain T at higher levels. To determine whether testosterone underlies the trade–off between mating and parental effort, we treated male dark–eyed juncos (Junco hyemalis) with exogenous T and compared the reproductive success (RS) of T–treated males (T–males) to that of controls. T–males had lower apparent annual RS than controls, probably because elevated T reduced parental care. Nevertheless, annual genetic RS of the treatment groups was similar because (i) T–males suffered fewer losses in genetic RS due to extra–pair fertilizations (EPFs), and (ii) T–males gained more genetic RS through their own EPFs. This is the first hormonal manipulation of an avian phenotype shown to have influenced male RS through EPFs. Together with other studies, it suggests that testosterone may have mediated the evolution of inter– and intraspecific differences in allocation of reproductive effort to mate attraction and parental care.

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.

Physiological Effects on Demography: A Long‐Term Experimental Study of Testosterone’s Effects on Fitness

Understanding physiological and behavioral mechanisms underlying the diversity of observed life‐history strategies is challenging because of difficulties in obtaining long‐term measures of fitness and in relating fitness to these mechanisms. We evaluated effects of experimentally elevated testosterone on male fitness in a population of dark‐eyed juncos studied over nine breeding seasons using a demographic modeling approach. Elevated levels of testosterone decreased survival rates but increased success of producing extra‐pair offspring. Higher overall fitness for testosterone‐treated males was unexpected and led us to consider indirect effects of testosterone on offspring and females. Nest success was similar for testosterone‐treated and control males, but testosterone‐treated males produced smaller offspring, and smaller offspring had lower postfledging survival. Older, more experienced females preferred to mate with older males and realized higher reproductive success when they did so. Treatment of young males increased their ability to attract older females yet resulted in poor reproductive performance. The higher fitness of testosterone‐treated males in the absence of a comparable natural phenotype suggests that the natural phenotype may be constrained. If this phenotype were to arise, the negative social effects on offspring and mates suggest that these effects might prevent high‐testosterone phenotypes from spreading in the population.

CORRELATIONAL SELECTION LEADS TO GENETIC INTEGRATION OF BODY SIZE AND AN ATTRACTIVE PLUMAGE TRAIT IN DARK‐EYED JUNCOS

Abstract When a traits effect on fitness depends on its interaction with other traits, the resultant selection is correlational and may lead to the integration of functionally related traits. In relation to sexual selection, when an ornamental trait interacts with phenotypic quality to determine mating success, correlational sexual selection should generate genetic correlations between the ornament and quality, leading to the evolution of honest signals. Despite its potential importance in the evolution of signal honesty, correlational sexual selection has rarely been measured in natural populations. In the dark‐eyed junco (Junco hyemalis), males with experimentally elevated values of a plumage trait (whiteness in the tail or “tail white”) are more attractive to females and dominant in aggressive encounters over resources. We used restricted maximum‐likelihood analysis of a long‐term dataset to measure the heritability of tail white and two components of body size (wing length and tail length), as well as genetic correlations between pairs of these traits. We then used multiple regression to assess directional, quadratic, and correlational selection as they acted on tail white and body size via four components of lifetime fitness (juvenile and adult survival, mating success, and fecundity). We found a positive genetic correlation between tail white and body size (as measured by wing length), which indicates past correlational selection. Correlational selection, which was largely due to sexual selection on males, was also found to be currently acting on the same pair of traits. Larger males with whiter tails sired young with more females, most likely due to a combination of female choice, which favors males with whiter tails, and male‐male competition, which favors both tail white and larger body size. To our knowledge, this is the first study to show both genetic correlations between sexually selected traits and currently acting correlational sexual selection, and we suggest that correlational sexual selection frequently may be an important mechanism for maintaining the honesty of sexual signals.

Using parasites to infer host population history: a new rationale for parasite conservation

Only one of the 5000 extant louse species (Phthiraptera) and no species of flea (Siphonaptera), parasitic helminth (Platyhelminthes), parasitic nematode (Nemata), mite, or tick (Acari) is listed as threatened by the IUCN, despite impassioned pleas for parasite conservation beginning more than a decade ago. Although they should be conserved for their own sake, past arguments, highlighting the intrinsic and utilitarian value of parasites, have not translated into increased attention by scientists or conservation managers, at least by the standard of listing for protection. Here, the use of estimated genealogies and population genetic patterns of parasites to illuminate their hosts’ evolutionary and demographic history is advocated. Parasite DNA generally evolves more rapidly than their hosts’, which renders it an underexploited resource for conservation biologists, particularly in cases where the hosts’ genealogy or degree of population genetic structure is difficult to measure directly. Moreover, parasite gene flow may occur during host dispersal irrespective of host gene flow, revealing host movement through space and time. Parasite ecology and evolution may thus become another tool for the management of endangered vertebrate populations. This will result in the recognition of new host records, parasite species and cryptic lineages, which will help lift the veil of ignorance with respect to parasite biodiversity.

Co‐phylogeography and comparative population genetics of the threatened Galápagos hawk and three ectoparasite species: ecology shapes population histories within parasite communities

Comparative microevolutionary studies of multiple parasites occurring on a single host species can help shed light on the processes underlying parasite diversification. We compared the phylogeographical histories, population genetic structures and population divergence times of three co‐distributed and phylogenetically independent ectoparasitic insect species, including an amblyceran and an ischnoceran louse (Insecta: Phthiraptera), a hippoboscid fly (Insecta: Diptera) and their endemic avian host in the Galápagos Islands. The Galápagos hawk (Aves: Falconiformes: Buteo galapagoensis) is a recently arrived endemic lineage in the Galápagos Islands and its island populations are diverging evolutionarily. Each parasite species differed in relative dispersal ability and distribution within the host populations, which allowed us to make predictions about their degree of population genetic structure and whether they tracked host gene flow and colonization history among islands. To control for DNA region in comparisons across these phylogenetically distant taxa, we sequenced ~1 kb of homologous mitochondrial DNA from samples collected from all island populations of the host. Remarkably, the host was invariant across mitochondrial regions that were comparatively variable in each of the parasite species, to degrees consistent with differences in their natural histories. Differences in these natural history traits were predictably correlated with the evolutionary trajectories of each parasite species, including rates of interisland gene flow and tracking of hosts by parasites. Congruence between the population structures of the ischnoceran louse and the host suggests that the ischnoceran may yield insight into the cryptic evolutionary history of its endangered host, potentially aiding in its conservation management.

Adult Survival and Imperfect Assessment of Parentage: Effects on Male Parenting Decisions

Applications of molecular methods to assess parentage have revealed that the distribution of reproductive success among individuals often differs, sometimes dramatically, from expectation based on observation of behavioral association. Much theory exists on whether and when males should reduce parental care in response to level of paternity. Life‐history theory predicts that trade‐offs in reproductive effort should be influenced by adult survival. We used a dynamic programming approach to address how level of paternity, ability to assess paternity, and adult survival rate interact to affect male tolerance of reduced parentage in a given brood. Adult survival has the greatest influence on male decisions such that, for any given cost of reproduction and value of male care, tolerance of extrapair fertilizations (EPFs) decreases as adult survival increases. An unexpected result of these models is that an optimal response also depends on a males ability to predict probability of parentage (i.e., uncertainty). These models better characterize the nature of paternity uncertainty and its effect on EPF tolerance than have previous models and add to our understanding of the complex relationship between uncertainty, mating strategies, and adult survival.

Confirmation of cooperative polyandry in the Galapagos hawk (Buteo galapagoensis)

Without genetic tests of parentage, descriptions of mating systems must be considered hypothetical. Here we confirm the existence of cooperative polyandry in the Galapagos hawk (Buteo galapagoensis) using multilocus minisatellite DNA fingerprinting. In this species, breeding groups consist of one adult female and from one to eight males (the modal number of males is two). In polyandrous groups, all males copulate with the female and participate in the provisioning of the young. DNA samples from 66 individual hawks from ten breeding groups on the island of Santiago revealed mixed paternity in most groups. Multiple paternity was detected in five of six groups that produced two chicks in one breeding attempt (year). In addition, different males sired young in consecutive years in five of six groups in which male group membership was constant. Patterns of paternity suggest that reproductive success was randomly distributed among males within groups, with males apparently having equivalent probabilities of siring each young. Analysis of genetic similarity indicates that males within groups were typically not close relatives These results demonstrate that the mating system of the Galapagos hawk is polyandrous, with relatively egalitarian relations among unrelated males belonging to the same breeding group.

Multiple Lineages of Avian Malaria Parasites (Plasmodium) in the Galapagos Islands and Evidence for Arrival via Migratory Birds

Haemosporidian parasites in the genus Plasmodium were recently detected through molecular screening in the Galapagos Penguin (Spheniscus mendiculus). We summarized results of an archipelago-wide screen of 3726 endemic birds representing 22 species for Plasmodium spp. through a combination of molecular and microscopy techniques. Three additional Plasmodium lineages were present in Galapagos. Lineage A-infected penguins, Yellow Warblers (Setophaga petechia aureola), and one Medium Ground Finch (Geospiza fortis) and was detected at multiple sites in multiple years [corrected]. The other 3 lineages were each detected at one site and at one time; apparently, they were transient infections of parasites not established on the archipelago. No gametocytes were found in blood smears of infected individuals; thus, endemic Galapagos birds may be dead-end hosts for these Plasmodium lineages. Determining when and how parasites and pathogens arrive in Galapagos is key to developing conservation strategies to prevent and mitigate the effects of introduced diseases. To assess the potential for Plasmodium parasites to arrive via migratory birds, we analyzed blood samples from 438 North American breeding Bobolinks (Dolichonyx oryzivorus), the only songbird that regularly migrates through Galapagos. Two of the ephemeral Plasmodium lineages (B and C) found in Galapagos birds matched parasite sequences from Bobolinks. Although this is not confirmation that Bobolinks are responsible for introducing these lineages, evidence points to higher potential arrival rates of avian pathogens than previously thought. Linajes Múltiples de Parásitos de Malaria Aviar (Plasmodium) en las Islas Galápagos y Evidencia de su Arribo por Medio de Aves Migratorias.