Diversity and prevalence of zoonotic infections at the animal-human interface of primate trafficking in Peru.

Wildlife trafficking creates favorable scenarios for intra- and inter-specific interactions that can lead to parasite spread and disease emergence. Among the fauna affected by this activity, primates are relevant due to their potential to acquire and share zoonoses - infections caused by parasites that can spread between humans and other animals. Though it is known that most primate parasites can affect multiple hosts and that many are zoonotic, comparative studies across different contexts for animal-human interactions are scarce. We conducted a multi-parasite screening targeting the detection of zoonotic infections in wild-caught monkeys in nine Peruvian cities across three contexts: captivity (zoos and rescue centers, n = 187); pet (households, n = 69); and trade (trafficked or recently confiscated, n = 132). We detected 32 parasite taxa including mycobacteria, simian foamyvirus, bacteria, helminths, and protozoa. Monkeys in the trade context had the highest prevalence of hemoparasites (including Plasmodium malariae/brasilianum, Trypanosoma cruzi, and microfilaria) and enteric helminths and protozoa were less common in pet monkeys. However, parasite communities showed overall low variation between the three contexts. Parasite richness (PR) was best explained by host genus and the city where the animal was sampled. Squirrel (genus Saimiri) and wooly (genus Lagothrix) monkeys had the highest PR, which was ~2.2 times the PR found in tufted capuchins (genus Sapajus) and tamarins (genus Saguinus/Leontocebus) in a multivariable model adjusted for context, sex, and age. Our findings illustrate that the threats of wildlife trafficking to One Health encompass exposure to multiple zoonotic parasites well-known to cause disease in humans, monkeys, and other species. We demonstrate these threats continue beyond the markets where wildlife is initially sold; monkeys trafficked for the pet market remain a reservoir for and contribute to the translocation of zoonotic parasites to households and other captive facilities where contact with humans is frequent. Our results have practical applications for the healthcare of rescued monkeys and call for urgent action against wildlife trafficking and ownership of monkeys as pets.

Composition and function of the Galapagos penguin gut microbiome vary with age, location, and a putative bacterial pathogen.

Microbial colonization plays a direct role in host health. Understanding the ecology of the resident microbial community for a given host species is thus an important step for detecting population vulnerabilities like disease. However, the idea of integrating microbiome research into conservation is still relatively new, and wild birds have received less attention in this field than mammals or domesticated animals. Here we examine the composition and function of the gut microbiome of the endangered Galapagos penguin (Spheniscus mendiculus) with the goals of characterizing the normal microbial community and resistome, identifying likely pathogens, and testing hypotheses of structuring forces for this community based on demographics, location, and infection status. We collected fecal samples from wild penguins in 2018 and performed 16S rRNA gene sequencing and whole genome sequencing (WGS) on extracted DNA. 16S sequencing revealed that the bacterial phyla Fusobacteria, Epsilonbacteraeota, Firmicutes, and Proteobacteria dominate the community. Functional pathways were computed from WGS data, showing genetic functional potential primarily focused on metabolism-amino acid metabolism, carbohydrate metabolism, and energy metabolism are the most well-represented functional groups. WGS samples were each screened for antimicrobial resistance, characterizing a resistome made up of nine antibiotic resistance genes. Samples were screened for potential enteric pathogens using virulence factors as indicators; Clostridium perfringens was revealed as a likely pathogen. Overall, three factors appear to be shaping the alpha and beta diversity of the microbial community: penguin developmental stage, sampling location, and C. perfringens. We found that juvenile penguins have significantly lower alpha diversity than adults based on three metrics, as well as significantly different beta diversity. Location effects are minimal, but one site has significantly lower Shannon diversity than the other primary sites. Finally, when samples were grouped by C. perfringens virulence factors, we found dramatic changes in beta diversity based on operational taxonomic units, protein families, and functional pathways. This study provides a baseline microbiome for an endangered species, implicates both penguin age and the presence of a potential bacterial pathogen as primary factors associated with microbial community variance, and reveals widespread antibiotic resistance genes across the population.

Contaminations contaminate common databases.

The polymerase chain reaction (PCR) is a very powerful method to detect and identify pathogens. The high sensitivity of the method, however, comes with a cost; any of the millions of artificial DNA copies generated by PCR can serve as a template in a following experiment. If not identified as contaminations, these may result in erroneous conclusions on the occurrence of the pathogen, thereby inflating estimates of host range and geographic distribution. In the present paper, we evaluate whether several published records of avian haemosporidian parasites, in either unusual host species or geographical regions, might stem from PCR contaminations rather than novel biological findings. The detailed descriptions of these cases are shedding light upon the steps in the work process that might lead to PCR contaminations. By increasing the awareness of this problem, it will aid in developing procedures that keep these to a minimum. The examples in the present paper are from haemosporidians of birds, however the problem of contaminations and suggested actions should apply generally to all kinds of PCR-based identifications, not just of parasites and pathogens.

Trypanosomatids Detected in the Invasive Avian Parasite Philornis downsi (Diptera: Muscidae) in the Galapagos Islands.

Alien insect species may present a multifaceted threat to ecosystems into which they are introduced. In addition to the direct damage they may cause, they may also bring novel diseases and parasites and/or have the capacity to vector microorganisms that are already established in the ecosystem and are causing harm. Damage caused by ectoparasitic larvae of the invasive fly, Philornis downsi (Dodge and Aitken) to nestlings of endemic birds in the Galapagos Islands is well documented, but nothing is known about whether this fly is itself associated with parasites or pathogens. In this study, diagnostic molecular methods indicated the presence of insect trypanosomatids in P. downsi; to our knowledge, this is the first record of insect trypanosomatids associated with Philornis species. Phylogenetic estimates and evolutionary distances indicate these species are most closely related to the Crithidia and Blastocrithidia genera, which are not currently reported in the Galapagos Islands. The prevalence of trypanosomatids indicates either P. downsi arrived with its own parasites or that it is a highly suitable host for trypanosomatids already found in the Galapagos Islands, or both. We recommend further studies to determine the origin of the trypanosomatid infections to better evaluate threats to endemic fauna of the Galapagos Islands.

Assessing the blood meal hosts of Culex quinquefasciatus and Aedes taeniorhynchus in Isla Santa Cruz, Galápagos.

BACKGROUND: Blood meal host selection by mosquito vectors is an important component in understanding disease dynamics of pathogens that threaten endemic fauna in isolated islands such as Galápagos. Research on the feeding behavior of mosquitoes can provide clues to the hosts and vectors involved in disease transmission. This information is particularly critical for endemic wildlife fauna in island systems that have evolved without resistance to novel diseases such as avian malaria. The aims of this study were to determine the blood-feeding patterns of two species of mosquitoes found in Galápagos and discuss how their feeding behavior may influence the transmission of pathogens such as avian malaria. METHODS: In the summer of 2015, we sampled two mosquito species (Aedes taeniorhynchus and Culex quinquefasciatus) across 18 different sites on Isla Santa Cruz, which is the second largest island in Galápagos and has the largest human population. We trapped mosquitoes using CDC light traps and CDC gravid traps and identified sources of blood meals for engorged mosquitoes by sequencing a portion of the vertebrate mitochondrial cytochrome b gene. RESULTS: Out of 947 female mosquitoes captured, 320 were blood-fed, and PCR amplifications were successful for 301 of the blood meals. Results revealed that both Aedes taeniorhynchus and Culex quinquefasciatus feed from a variety of vertebrate taxa, numerically dominated by humans on Isla Santa Cruz. CONCLUSIONS: The high proportion of mammalian blood meals could represent locally available and abundant hosts on Santa Cruz. However, host surveys and estimates of relative abundances of vertebrate species will need to accompany mosquito trapping studies on non-inhabited and inhabited islands in Galápagos to further validate this.

A multiyear survey of helminths from wild saddleback (Leontocebus weddelli) and emperor (Saguinus imperator) tamarins.

The establishment of baseline data on parasites from wild primates is essential to understand how changes in habitat or climatic disturbances will impact parasite-host relationships. In nature, multiparasitic infections of primates usually fluctuate temporally and seasonally, implying that the acquisition of reliable data must occur over time. Individual parasite infection data from two wild populations of New World primates, the saddleback (Leontocebus weddelli) and emperor (Saguinus imperator) tamarin, were collected over 3 years to establish baseline levels of helminth prevalence and parasite species richness (PSR). Secondarily, we explored variation in parasite prevalence across age and sex classes, test nonrandom associations of parasite co-occurrence, and assess the relationship between group size and PSR. From 288 fecal samples across 105 individuals (71 saddleback and 34 emperor tamarins), 10 parasite taxa were identified by light microscopy following centrifugation and ethyl-acetate sedimentation. Of these taxa, none were host-specific, Dicrocoeliidae and Cestoda prevalences differed between host species, Prosthenorchis and Strongylida were the most prevalent. Host age was positively associated with Prosthenorchis ova and filariform larva, but negatively with cestode and the Rhabditoidea ova. We detected no differences between expected and observed levels of co-infection, nor between group size and parasite species richness over 30 group-years. Logistic models of individual infection status did not identify a sex bias; however, age and species predicted the presence of four and three parasite taxa, respectively, with saddleback tamarins exhibiting higher PSR. Now that we have reliable baseline data for future monitoring of these populations, next steps involve the molecular characterization of these parasites, and exploration of linkages with health parameters.

Haemosporidian parasite community in migrating bobolinks on the Galapagos Islands.

Bobolinks (Dolichonyx oryzivorus) migrate from their breeding grounds in North America to their wintering grounds in South America during the fall each year. A small number of Bobolinks stop temporarily in Galapagos, and potentially carry parasites. On the North American breeding grounds, Bobolinks carry a least two of the four Plasmodium lineages recently detected in resident Galapagos birds. We hypothesized that Bobolinks carried these parasites to Galapagos, where they were bitten by mosquitoes that then transmitted the parasites to resident birds. The haemosporidian parasite community in 44% of the Bobolinks we captured was consistent with those on their breeding grounds. However, the lineages were not those found in Galapagos birds. Our results provide a parasite community key for future monitoring.

The influence of ecological factors on mosquito abundance and occurrence in Galápagos.

We sampled mosquitoes across 18 sites established at different elevations and stretching from the north to the south of Isla Santa Cruz, Galápagos. Two commonly occurring species, Ae. taeniorhynchus and Cx. quinquefasciatus, were collected along with environmental variables characteristic of the trapping sites to assess their influence on mosquito abundance and occurrence in the dry season of 2015. We captured Ae. taeniorhynchus at 14 out of 18 sites and Cx. quinquefasciatus at low and high elevation sites on Santa Cruz. We utilized two generalized linear models; the first assessed the influence of environmental variables on abundances of Ae. taeniorhynchus and the second assessed the influence of these variables on the presence of Cx. quinquefasciatus. Populations of both mosquito species declined with elevation. Rainfall data were limited, as we sampled during the dry season of 2015. Distance to mangroves and maximum humidity were significant in influencing the abundance of Ae. taeniorhynchus, while maximum humidity was found to significantly influence the presence of Cx. quinquefasciatus. Both species occurred in sites where temperature, precipitation, and humidity should allow for mosquito development as well as parasitic development of the protozoan parasites that cause avian malaria. Further research involving year-round sampling of mosquitoes and accompanying meteorological data as well as experimental studies on vector competence are required to understand disease dynamics of parasites such as avian malaria in Galápagos.

Patterns of Exposure of Carnivores to Selected Pathogens in the Betampona Natural Reserve Landscape, Madagascar.

Carnivores of Madagascar are at increased risk of extinction due to anthropogenic loss of habitat, hunting, and interactions with introduced carnivores. Interactions between introduced and native animals also present the potential for introduction of pathogens into new geographic areas or host species. Here, we provide serologic data regarding pathogen exposure of domestic and native carnivores from the Betampona Natural Reserve Landscape, a protected area in eastern Madagascar. For the Eupleridae, we found limited evidence of exposure to viruses from domestic animals but greater prevalence for Toxoplasma gondii (39%) and Leptospira interrogans (40%). We also evaluated the associations between the presence of antibodies to selected pathogens and the demographic and spatial variables. We showed that individual characteristics such as sex and species were associated with exposure to T. gondii but not to L. interrogans or canine parvovirus (CPV). Finally, we investigated the spatial structure of pathogen exposure in Betampona and found no evidence of spatial structuring, indicating the absence of hotspots and agent-free refugia for T. gondii, L. interrogans, and CPV in the protected area. Our results may be useful for assessing and monitoring disease risk and for formulating control strategies to minimize the negative impact of exotic species on the endemic carnivores of Madagascar.

Landscape genomics: natural selection drives the evolution of mitogenome in penguins.

BACKGROUND: Mitochondria play a key role in the balance of energy and heat production, and therefore the mitochondrial genome is under natural selection by environmental temperature and food availability, since starvation can generate more efficient coupling of energy production. However, selection over mitochondrial DNA (mtDNA) genes has usually been evaluated at the population level. We sequenced by NGS 12 mitogenomes and with four published genomes, assessed genetic variation in ten penguin species distributed from the equator to Antarctica. Signatures of selection of 13 mitochondrial protein-coding genes were evaluated by comparing among species within and among genera (Spheniscus, Pygoscelis, Eudyptula, Eudyptes and Aptenodytes). The genetic data were correlated with environmental data obtained through remote sensing (sea surface temperature [SST], chlorophyll levels [Chl] and a combination of SST and Chl [COM]) through the distribution of these species. RESULTS: We identified the complete mtDNA genomes of several penguin species, including ND6 and 8 tRNAs on the light strand and 12 protein coding genes, 14 tRNAs and two rRNAs positioned on the heavy strand. The highest diversity was found in NADH dehydrogenase genes and the lowest in COX genes. The lowest evolutionary divergence among species was between Humboldt (Spheniscus humboldti) and Galapagos (S. mendiculus) penguins (0.004), while the highest was observed between little penguin (Eudyptula minor) and Adélie penguin (Pygoscelis adeliae) (0.097). We identified a signature of purifying selection (Ka/Ks < 1) across the mitochondrial genome, which is consistent with the hypothesis that purifying selection is constraining mitogenome evolution to maintain Oxidative phosphorylation (OXPHOS) proteins and functionality. Pairwise species maximum-likelihood analyses of selection at codon sites suggest positive selection has occurred on ATP8 (Fixed-Effects Likelihood, FEL) and ND4 (Single Likelihood Ancestral Counting, SLAC) in all penguins. In contrast, COX1 had a signature of strong negative selection. ND4 Ka/Ks ratios were highly correlated with SST (Mantel, p-value: 0.0001; GLM, p-value: 0.00001) and thus may be related to climate adaptation throughout penguin speciation. CONCLUSIONS: These results identify mtDNA candidate genes under selection which could be involved in broad-scale adaptations of penguins to their environment. Such knowledge may be particularly useful for developing predictive models of how these species may respond to severe climatic changes in the future.

Mode and Rate of Evolution of Haemosporidian Mitochondrial Genomes: Timing the Radiation of Avian Parasites.

Haemosporidians are a diverse group of vector-borne parasitic protozoa that includes the agents of human malaria; however, most of the described species are found in birds and reptiles. Although our understanding of these parasites' diversity has expanded by analyses of their mitochondrial genes, there is limited information on these genes' evolutionary rates. Here, 114 mitochondrial genomes (mtDNA) were studied from species belonging to four genera: Leucocytozoon, Haemoproteus, Hepatocystis, and Plasmodium. Contrary to previous assertions, the mtDNA is phylogenetically informative. The inferred phylogeny showed that, like the genus Plasmodium, the Leucocytozoon and Haemoproteus genera are not monophyletic groups. Although sensitive to the assumptions of the molecular dating method used, the estimated times indicate that the diversification of the avian haemosporidian subgenera/genera took place after the Cretaceous-Paleogene boundary following the radiation of modern birds. Furthermore, parasite clade differences in mtDNA substitution rates and strength of negative selection were detected. These differences may affect the biological interpretation of mtDNA gene lineages used as a proxy to species in ecological and parasitological investigations. Given that the mitochondria are critically important in the parasite life cycle stages that take place in the vector and that the transmission of parasites belonging to particular clades has been linked to specific insect families/subfamilies, this study suggests that differences in vectors have affected the mode of evolution of haemosporidian mtDNA genes. The observed patterns also suggest that the radiation of haemosporidian parasites may be the result of community-level evolutionary processes between their vertebrate and invertebrate hosts.

The distribution of mosquitoes across an altitudinal gradient in the Galapagos Islands.

An avian malaria parasite (genus Plasmodium) has been detected consistently in the Galapagos Penguin (Spheniscus mendiculus) and less frequently in some passerines. We sampled three resident mosquito species (Aedes taeniorhynchus, Culex quinquefasciatus, and Aedes aegypti) using CDC light and gravid traps on three islands in 2012, 2013, and 2014. We sampled along altitudinal gradients to ask whether there are mosquito-free refugia at higher elevations as there are in Hawaii. We captured both Ae. taeniorhynchus and Cx. quinquefasciatus at all sites. However, abundances differed across islands and years and declined significantly with elevation. Aedes aegypti were scarce and limited to areas of human inhabitation. These results were corroborated by two negative binomial regression models which found altitude, year, trap type, and island as categorized by human inhabitation to be significant factors influencing the distributions of both Ae. taeniorhynchus and Cx. quinquefasciatus. Annual differences at the highest altitudes in Isabela and Santa Cruz indicate the lack of a stable highland refuge if either species is found to be a major vector of a parasite, such as avian malaria in Galapagos. Further work is needed to confirm the vector potential of both species to understand the disease dynamics of avian malaria in Galapagos.

Interactions Between Carnivores in Madagascar and the Risk of Disease Transmission.

Introduced carnivores exert considerable pressure on native predators through predation, competition and disease transmission. Recent research shows that exotic carnivores negatively affect the distribution and abundance of the native and endangered carnivores of Madagascar. In this study, we provide information about the frequency and distribution of interactions between exotic (dogs and cats) and native carnivores (Eupleridae) in the Betampona Natural Reserve (BNR), Madagascar, using noninvasive camera trap surveys. Domestic dogs (Canis familiaris) were the most frequently detected carnivore species within the BNR, and we found that indirect interactions between exotic and native carnivores were frequent (n = 236). Indirect interactions were more likely to occur near the research station (incidence rate ratio = 0.91), which may constitute a disease transmission hot spot for carnivores at BNR. The intervals between capture of native and exotic carnivores suggest that there is potential for pathogen transmission between species in BNR. These capture intervals were significantly shorter near the edge of the reserve (P = 0.04). These data could be used to implement biosecurity measures to monitor interactions and prevent disease transmission between species at the domestic animal and wildlife interface.

Chronic Plasmodium brasilianum infections in wild Peruvian tamarins.

There is an increased interest in potential zoonotic malarias. To date, Plasmodium malariae that infects humans remains indistinguishable from Plasmodium brasilianum, which is widespread among New World primates. Distributed throughout tropical Central and South America, the Callitrichidae are small arboreal primates in which detection of natural Plasmodium infection has been extremely rare. Most prior screening efforts have been limited to small samples, the use of low-probability detection methods, or both. Rarely have screening efforts implemented a longitudinal sampling design. Through an annual mark-recapture program of two sympatric callitrichids, the emperor (Saguinus imperator) and saddleback (Saguinus fuscicollis) tamarins, whole blood samples were screened for Plasmodium by microscopy and nested PCR of the cytochrome b gene across four consecutive years (2012-2015). Following the first field season, approximately 50% of the samples collected each subsequent year were from recaptured individuals. In particular, out of 245 samples from 129 individuals, 11 samples from 6 individuals were positive for Plasmodium, and all but one of these infections was found in S. imperator. Importantly, the cytochrome b sequences were 100% identical to former isolates of P. malariae from humans and P. brasilianum from Saimiri sp. Chronic infections were detected as evidenced by repeated infections (7) from two individuals across the 4-year study period. Furthermore, 4 of the 5 infected emperor tamarins were part of a single group spanning the entire study period. Overall, the low prevalence reported here is consistent with previous findings. This study identifies two new natural hosts for P. brasilianum and provides evidence in support of chronic infections in wildlife populations. Given that callitrichids are often found in mixed-species associations with other primates and can be resilient to human-disturbed environments, they could contribute to the maintenance of P. malariae populations if future work provides entomological and epidemiological evidence indicating human zoonotic infections.

From Galapagos doves to passerines: Spillover of Haemoproteus multipigmentatus.

Haemoproteus (Haemoproteus) multipigmentatus, a haemosporidian parasite thought to be specific to columbiform birds, was detected in passeriform birds on Santiago Island in the Galapagos archipelago. We surveyed birds along an altitudinal gradient on the islands of Santa Cruz, Isabela and Santiago between June 2013 and July 2015. Molecular screening of 2254 individuals from 25 species of endemic and introduced birds revealed clusters of passerine birds positive for H. multipigmentatus on Santiago Island that coincide with captures of Galapagos doves at sampled sites. Of 507 individuals from 10 species of endemic passerines sampled on Santiago, 58 individuals from 6 species were found positive (11% prevalence). However, no gametocytes were found in the blood smears of positive passerines, suggesting that these species are not competent hosts for the parasite. All 31 doves captured were positive and gametocytes were found upon microscopic examination of all thin blood smears (averaging 357 gametocytes per 10,000 erythrocytes). These findings indicate parasite spillover from doves to passerines, but that passerines are possibly not competent hosts for further parasite transmission. The endemic Galapagos dove acts as a reservoir host for the introduced H. multipigmentatus, however the effect of this parasite on passerines has not been studied. We report on these findings because parasites can have large effects on individual host populations and on the ecology of a community, but may go undetected.

A genetic signature of the evolution of loss of flight in the Galapagos cormorant.

We have a limited understanding of the genetic and molecular basis of evolutionary changes in the size and proportion of limbs. We studied wing and pectoral skeleton reduction leading to flightlessness in the Galapagos cormorant (Phalacrocorax harrisi). We sequenced and de novo assembled the genomes of four cormorant species and applied a predictive and comparative genomics approach to find candidate variants that may have contributed to the evolution of flightlessness. These analyses and cross-species experiments in Caenorhabditis elegans and in chondrogenic cell lines implicated variants in genes necessary for transcriptional regulation and function of the primary cilium. Cilia are essential for Hedgehog signaling, and humans affected by skeletal ciliopathies suffer from premature bone growth arrest, mirroring skeletal features associated with loss of flight.

Host sympatry and body size influence parasite straggling rate in a highly connected multihost, multiparasite system.

Parasite lineages commonly diverge when host lineages diverge. However, when large clades of hosts and parasites are analyzed, some cases suggest host switching as another major diversification mechanism. The first step in host switching is the appearance of a parasite on an atypical host, or "straggling." We analyze the conditions associated with straggling events. We use five species of colonially nesting seabirds from the Galapagos Archipelago and two genera of highly specific ectoparasitic lice to examine host switching. We use both genetic and morphological identification of lice, together with measurements of spatial distribution of hosts in mixed breeding colonies, to test: (1) effects of local host community composition on straggling parasite identity; (2) effects of relative host density within a mixed colony on straggling frequency and parasite species identity; and (3) how straggling rates are influenced by the specifics of louse attachment. Finally, we determine whether there is evidence of breeding in cases where straggling adult lice were found, which may indicate a shift from straggling to the initial stages of host switching. We analyzed more than 5,000 parasite individuals and found that only ~1% of lice could be considered stragglers, with ~5% of 436 host individuals having straggling parasites. We found that the presence of the typical host and recipient host in the same locality influenced straggling. Additionally, parasites most likely to be found on alternate hosts are those that are smaller than the typical parasite of that host, implying that the ability of lice to attach to the host might limit host switching. Given that lice generally follow Harrison's rule, with larger parasites on larger hosts, parasites infecting the larger host species are less likely to successfully colonize smaller host species. Moreover, our study supports the general perception that successful colonization of a novel host is extremely rare, as we found only one nymph of a straggling species, which may indicate successful reproduction.

Temporal and demographic blood parasite dynamics in two free-ranging neotropical primates.

Parasite-host relationships are influenced by several factors intrinsic to hosts, such as social standing, group membership, sex, and age. However, in wild populations, temporal variation in parasite distributions and concomitant infections can alter these patterns. We used microscropy and molecular methods to screen for naturally occurring haemoparasitic infections in two Neotropical primate host populations, the saddleback (Leontocebus weddelli) and emperor (Saguinus imperator) tamarin, in the lowland tropical rainforests of southeastern Peru. Repeat sampling was conducted from known individuals over a three-year period to test for parasite-host and parasite-parasite associations. Three parasites were detected in L. weddelli including Trypanosoma minasense, Mansonella mariae, and Dipetalonema spp., while S. imperator only hosted the latter two. Temporal variation in prevalence was observed in T. minasense and Dipetalonema spp., confirming the necessity of a multi-year study to evaluate parasite-host relationships in this system. Although callitrichids display a distinct reproductive dominance hierarchy, characterized by single breeding females that typically mate polyandrously and can suppress the reproduction of subdominant females, logistic models did not identify sex or breeding status as determining factors in the presence of these parasites. However, age class had a positive effect on infection with M. mariae and T. minasense, and adults demonstrated higher parasite species richness than juveniles or sub-adults across both species. Body weight had a positive effect on the presence of Dipetalonema spp. The inclusion of co-infection variables in statistical models of parasite presence/absence data improved model fit for two of three parasites. This study verifies the importance and need for broad spectrum and long-term screening of parasite assemblages of natural host populations.

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

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.

Lineage sorting in multihost parasites: Eidmanniella albescens and Fregatiella aurifasciata on seabirds from the Galapagos Islands.

Parasites comprise a significant percentage of the biodiversity of the planet and are useful systems to test evolutionary and ecological hypotheses. In this study, we analyze the effect of host species identity and the immediate local species assemblage within mixed species colonies of nesting seabirds on patterns of genetic clustering within two species of multihost ectoparasitic lice. We use three genetic markers (one mitochondrial, COI, and two nuclear, EF1-α and wingless) and maximum likelihood phylogenetic trees to test whether (1) parasites show lineage sorting based on their host species; and (2) switching of lineages to the alternate host species depends on the immediate local species assemblage of individual hosts within a colony. Specifically, we examine the genetic structure of two louse species: Eidmanniella albescens, infecting both Nazca (Sula granti) and blue-footed boobies (Sula nebouxii), and Fregatiella aurifasciata, infecting both great (Fregata minor) and magnificent frigatebirds (Fregata magnificens). We found that host species identity was the only factor explaining the patterns of genetic structure in both parasites. In both cases, there is evident genetic differentiation depending on the host species. Thus, a revision of the taxonomy of these louse species is needed. One possible explanation of this pattern is extremely low louse migration rates between host species, perhaps influenced by fine-scale spatial separation of host species within mixed colonies, and low parasite infrapopulation numbers.