Iris I. Levin

A new Haemoproteus species (Haemosporida: Haemoproteidae) from the endemic Galapagos dove Zenaida galapagoensis, with remarks on the parasite distribution, vectors, and molecular diagnostics.

Abstract Haemoproteus (Haemoproteus) multipigmentatus n. sp. (Haemosporida, Haemoproteidae) was found in the endemic Galapagos dove Zenaida galapagoensis. It is described based on the morphology of its blood stages and segments of the mitochondrial cytochrome b gene, which can be used for molecular identification and diagnosis of this species. Haemoproteus multipigmentatus can be readily distinguished from all species of hemoproteids of the subgenus Haemoproteus, primarily due to numerous (approximately 40 on average) small pigment granules in its mature gametocytes. Illustrations of blood stages of the new species are given, and phylogenetic analysis identifies DNA lineages closely related to this parasite, which is prevalent in the Galapagos dove and also has been recorded in other species of Columbiformes in Mexico, Guatemala, and Peru, and so seems to be widespread in countries in the New World with warm climates. Cytochrome b lineages of H. multipigmentatus cluster with hippoboscid-transmitted lineages of Haemoproteus columbae. The same lineages of H. multipigmentatus were recorded in thoraxes of the hippoboscid fly Microlynchia galapagoensis, which likely is a natural vector of this parasite in Galapagos. Because different primers might amplify different parasites if they have a better match during a simultaneous infection, it is important that researchers standardize the genetic marker of choice for molecular typing of hemosporidian species. This study shows that more discussion among researchers is needed to clearly establish the sequence length and number of genes used for identification of hemosporidian parasites at different taxonomic levels. We point to the need of using both morphology and gene markers in studies of hemosporidian parasites, particularly in wildlife.

Hippoboscid-transmitted Haemoproteus parasites (Haemosporida) infect Galapagos Pelecaniform birds: Evidence from molecular and morphological studies, with a description of Haemoproteus iwa

Haemosporidian parasites are widely distributed and common parasites of birds, and the application of molecular techniques has revealed remarkable diversity among their lineages. Four haemosporidian genera infect avian hosts (Plasmodium, Haemoproteus, Leucocytozoon and Fallisia), and Haemoproteus is split into two sub-genera based on morphological evidence and phylogenetic support for two divergent sister clades. One clade (Haemoproteus (Parahaemoproteus)) contains parasites developing in birds belonging to several different orders, except pigeons and doves (Columbiformes), while the other (Haemoproteus (Haemoproteus)) has previously been shown to only infect dove hosts. Here we provide molecular and morphological identification of Haemoproteus parasites from several seabird species that are closely related to those found in dove hosts. We also document a deeply divergent clade with two haemosporidian lineages recovered primarily from frigatebirds (Fregatidae, Pelecaniformes) that is sister to the hippoboscid-(Hippoboscidae) transmitted dove parasites. One of the lineages in this new clade of parasites belongs to Haemoproteus iwa and is distributed in two species of frigatebird (Fregata) hosts from Hawaii, the Galapagos Islands, the eastern Pacific and throughout the Caribbean Basin. Haemosporidian parasites are often considered rare in seabirds due in part to the lack or low activity of some dipteran vectors (e.g., mosquitos, biting midges) in marine and coastal environments; however, we show that H. iwa is prevalent and is very likely vectored among frigatebirds by hippoboscid flies which are abundant on frigatebirds and other seabirds. This study supports the existence of two sister clades of avian Haemoproteus in accord with the subgeneric classification of avian hemoproteids. Description of H. iwa from Galapagos Fregata minor is given based on morphology of blood stages and segments of the mitochondrial cytochrome b gene, which can be used for identification. This study shows that hippoboscid flies warrant more attention as vectors of avian Haemoproteus spp., particularly in marine and coastal environments.

Novel Haemoproteus Species (Haemosporida: Haemoproteidae) from the Swallow-Tailed Gull (Lariidae), with Remarks On the Host Range of Hippoboscid-Transmitted Avian Hemoproteids

Abstract: Haemoproteus (Haemoproteus) jenniae n. sp. (Haemosporida: Haemoproteidae) is described from a Galapagos bird, the swallow-tailed gull Creagrus furcatus (Charadriiformes, Laridae), based on the morphology of its blood stages and segments of the mitochondrial cytochrome b (cyt b) gene. The most distinctive features of H. jenniae development are the circumnuclear gametocytes occupying all cytoplasmic space in infected erythrocytes and the presence of advanced, growing gametocytes in which the pellicle is closely appressed to the erythrocyte envelope but does not extend to the erythrocyte nucleus. This parasite is distinguishable from Haemoproteus larae, which produces similar gametocytes and parasitizes closely related species of Laridae. Haemoproteus jenniae can be distinguished from H. larae primarily due to (1) the predominantly amoeboid outline of young gametocytes, (2) diffuse macrogametocyte nuclei which do not possess distinguishable nucleoli, (3) the consistent size and shape of pigment granules, and (4) the absence of rod-like pigment granules from gametocytes. Additionally, fully-grown gametocytes of H. jenniae cause both the marked hypertrophy of infected erythrocytes in width and the rounding up of the host cells, which is not the case in H. larae. Phylogenetic analyses identified the DNA lineages that are associated with H. jenniae and showed that this parasite is more closely related to the hippoboscid-transmitted (Hippoboscidae) species than to the Culicoides spp.-transmitted (Ceratopogonidae) species of avian hemoproteids. Genetic divergence between morphologically well-differentiated H. jenniae and the hippoboscid-transmitted Haemoproteus iwa, the closely related parasite of frigatebirds (Fregatidae, Pelecaniformes), is only 0.6%; cyt b sequences of these parasites differ only by 1 base pair. This is the first example of such a small genetic difference in the cyt b gene between species of the subgenus Haemoproteus. In a segment of caseinolytic protease C gene (ClpC), genetic divergence is 4% between H. jenniae and H. iwa. This study corroborates the conclusion that hippoboscid-transmitted Haemoproteus parasites infect not only Columbiformes birds but also infect marine birds belonging to Pelecaniformes and Charadriiformes. We conclude that the vertebrate host range should be used cautiously in identification of subgenera of avian Haemoproteus species and that the phylogenies based on the cyt b gene provide evidence for determining the subgeneric position of avian hemoproteids.

Long-term isolation of a highly mobile seabird on the Galapagos

The Galapagos Islands are renowned for their high degree of endemism. Marine taxa inhabiting the archipelago might be expected to be an exception, because of their utilization of pelagic habitats—the dispersal barrier for terrestrial taxa—as foraging grounds. Magnificent frigatebirds (Fregata magnificens) have a highly vagile lifestyle and wide geographical distribution around the South and Central American coasts. Given the potentially high levels of gene flow among populations, the species provides a good test of the effectiveness of the Galapagos ecosystem in isolating populations of highly dispersive marine species. We studied patterns of genetic (mitochondrial DNA, microsatellites and nuclear introns) and morphological variation across the distribution of magnificent frigatebirds. Concordant with predictions from life-history traits, we found signatures of extensive gene flow over most of the range, even across the Isthmus of Panama, which is a major barrier to gene flow in other tropical seabirds. In contrast, individuals from the Galapagos were strongly differentiated from all conspecifics, and have probably been isolated for several hundred thousand years. Our finding is a powerful testimony to the evolutionary uniqueness of the taxa inhabiting the Galapagos archipelago and its associated marine ecosystems.

Philopatry drives genetic differentiation in an island archipelago: comparative population genetics of Galapagos Nazca boobies (Sula granti) and great frigatebirds (Fregata minor)

Seabirds are considered highly mobile, able to fly great distances with few apparent barriers to dispersal. However, it is often the case that seabird populations exhibit strong population genetic structure despite their potential vagility. Here we show that Galapagos Nazca booby (Sula granti) populations are substantially differentiated, even within the small geographic scale of this archipelago. On the other hand, Galapagos great frigatebird (Fregata minor) populations do not show any genetic structure. We characterized the genetic differentiation by sampling five colonies of both species in the Galapagos archipelago and analyzing eight microsatellite loci and three mitochondrial genes. Using an F-statistic approach on the multilocus data, we found significant differentiation between nearly all island pairs of Nazca booby populations and a Bayesian clustering analysis provided support for three distinct genetic clusters. Mitochondrial DNA showed less differentiation of Nazca booby colonies; only Nazca boobies from the island of Darwin were significantly differentiated from individuals throughout the rest of the archipelago. Great frigatebird populations showed little to no evidence for genetic differentiation at the same scale. Only two island pairs (Darwin – Wolf, N. Seymour – Wolf) were significantly differentiated using the multilocus data, and only two island pairs had statistically significant φST values (N. Seymour – Darwin, N. Seymour – Wolf) according to the mitochondrial data. There was no significant pattern of isolation by distance for either species calculated using both markers. Seven of the ten Nazca booby migration rates calculated between island pairs were in the south or southeast to north or northwest direction. The population differentiation found among Galapagos Nazca booby colonies, but not great frigatebird colonies, is most likely due to differences in natal and breeding philopatry.

Comparative host-parasite population genetic structures: obligate fly ectoparasites on Galapagos seabirds

Parasites often have shorter generation times and, in some cases, faster mutation rates than their hosts, which can lead to greater population differentiation in the parasite relative to the host. Here we present a population genetic study of two ectoparasitic flies, Olfersia spinifera and Olfersia aenescens compared with their respective bird hosts, great frigatebirds (Fregata minor) and Nazca boobies (Sula granti). Olfersia spinifera is the vector of a haemosporidian parasite, Haemoproteus iwa, which infects frigatebirds throughout their range. Interestingly, there is no genetic differentiation in the haemosporidian parasite across this range despite strong genetic differentiation between Galapagos frigatebirds and their non-Galapagos conspecifics. It is possible that the broad distribution of this one H. iwa lineage could be facilitated by movement of infected O. spinifera. Therefore, we predicted more gene flow in both fly species compared with the bird hosts. Mitochondrial DNA sequence data from three genes per species indicated that despite marked differences in the genetic structure of the bird hosts, gene flow was very high in both fly species. A likely explanation involves non-breeding movements of hosts, including movement of juveniles, and movement by adult birds whose breeding attempt has failed, although we cannot rule out the possibility that closely related host species may be involved.

Three decades of cultural evolution in Savannah sparrow songs

Cultural evolution can result in changes in the prevalence not only of different learned song types within bird populations but also of different segments within the song. Between 1980 and 2011, we examined changes within different segments of the single songs of male Savannah sparrows, Passerculus sandwichiensis, in an island population. Introductory notes did not change. The buzz segment showed similar stability; although a rare low-frequency variant appeared and then disappeared, the buzz segments from 1980 and 2011 were essentially identical. The middle segment, made up of discrete notes assembled into several types, was variable. However, the form of the middle segment did not affect fitness and may serve to denote individual identity. The terminal trill decreased steadily in frequency and duration over three decades. Longer trills were associated with lower reproductive success, suggesting that trill duration was under sexual selection. The notes sung between introductory notes were also associated with reproductive success. A high cluster sung in 1980e1982 disappeared altogether by 2011, and was gradually replaced by click trains, which were associated with greater reproductive success. During the final decade of the study, more clicks were added to click trains. Longer click trains, which may require vocal virtuosity and so indicate male quality, were also associated with greater reproductive success. Both trill duration and the number of clicks increased in variance during the three-decade span of the study. We suggest that such increases in variance might be a signature of directional cultural selection. Within the Savannah sparrow’s relatively short and simple learned song, cultural evolution appears to be mediated by different mechanisms for different song segments, perhaps because the segments convey different information.

Infection with Haemoproteus iwa affects vector movement in a hippoboscid fly—frigatebird system

Haemosporidian parasites, which require both a vertebrate and invertebrate host, are most commonly studied in the life stages occurring in the vertebrate. However, aspects of the vectors behaviour and biology can have profound effects on parasite dynamics. We explored the effects of a haemosporidian parasite, Haemoproteus iwa, on a hippoboscid fly vector, Olfersia spinifera. Olfersia spinifera is an obligate ectoparasite of the great frigatebird, Fregata minor, living among bird feathers for all of its adult life. This study examined the movements of O. spinifera between great frigatebird hosts. Movement, or host switching, was inferred by identifying host (frigatebird) microsatellite genotypes from fly bloodmeals that did not match the host from which the fly was collected. Such host switches were analysed using a logistic regression model, and the best‐fit model included the H. iwa infection status of the fly and the bird host sex. Uninfected flies were more likely to have a bird genotype in their bloodmeal that was different from their current hosts genotype (i.e. to have switched hosts) than infected flies. Flies collected from female birds were more likely to have switched hosts than those collected on males. Reduced movement of infected flies suggests that there may be a cost of parasitism for the fly. The effect of host sex is probably driven by differences in the sex ratio of bird hosts available to moving flies.

Local parasite lineage sharing in temperate grassland birds provides clues about potential origins of Galapagos avian Plasmodium

Abstract Oceanic archipelagos are vulnerable to natural introduction of parasites via migratory birds. Our aim was to characterize the geographic origins of two Plasmodium parasite lineages detected in the Galapagos Islands and in North American breeding bobolinks (Dolichonyx oryzivorus) that regularly stop in Galapagos during migration to their South American overwintering sites. We used samples from a grassland breeding bird assemblage in Nebraska, United States, and parasite DNA sequences from the Galapagos Islands, Ecuador, to compare to global data in a DNA sequence registry. Homologous DNA sequences from parasites detected in bobolinks and more sedentary birds (e.g., brown‐headed cowbirds Molothrus ater, and other co‐occurring bird species resident on the North American breeding grounds) were compared to those recovered in previous studies from global sites. One parasite lineage that matched between Galapagos birds and the migratory bobolink, Plasmodium lineage B, was the most common lineage detected in the global MalAvi database, matching 49 sequences from unique host/site combinations, 41 of which were of South American origin. We did not detect lineage B in brown‐headed cowbirds. The other Galapagos‐bobolink match, Plasmodium lineage C, was identical to two other sequences from birds sampled in California. We detected a close variant of lineage C in brown‐headed cowbirds. Taken together, this pattern suggests that bobolinks became infected with lineage B on the South American end of their migratory range, and with lineage C on the North American breeding grounds. Overall, we detected more parasite lineages in bobolinks than in cowbirds. Galapagos Plasmodium had similar host breadth compared to the non‐Galapagos haemosporidian lineages detected in bobolinks, brown‐headed cowbirds, and other grassland species. This study highlights the utility of global haemosporidian data in the context of migratory bird–parasite connectivity. It is possible that migratory bobolinks bring parasites to the Galapagos and that these parasites originate from different biogeographic regions representing both their breeding and overwintering sites.

Comparative Ectoparasite Loads of Five Seabird Species in the Galapagos Islands

Abstract: In this paper we describe the ectoparasitic lice (Insecta: Phthiraptera) found on 5 species of seabirds (magnificent frigatebird Fregata magnificens; great frigatebird Fregata minor; Nazca booby Sula granti; blue-footed booby Sula nebouxii; and red-footed booby Sula sula) on the Galapagos Archipelago. We found 9 species of ectoparasitic lice: 5 species of Pectinopygus ischnocerans, 1 infesting each host; 2 species of Colpocephalum amblyceran lice, 1 on each frigatebird species; and 2 shared amblycerans, Eidmanniella albescens (Piaget, 1880) found on Nazca and blue-footed boobies and Fregatiella aurifasciata (Kellogg, 1899) found on the 2 frigatebirds. We tested the relative importance and interactions of host sex, body size, host, island, host family, and breeding status and found that inter-island differences were the main predictors of prevalence and infestation intensity. These differences could be related to host density or weather, but further evidence is needed.