Pathogenicity of avian malaria in experimentally-infected Hawaii Amakihi

The introduction of avian malaria (Plasmodium relictum) and mosquitoes (Culex quinquefasciatus) to the Hawaiian Islands (USA) is believed to have played a major role in the decline and extinction of native Hawaiian honeycreepers (Drepanidinae). This introduced disease is thought to be one of the primary factors limiting recovery of honeycreepers at elevations below 1,200 m where native forest habitats are still relatively intact. One of the few remaining species of honeycreepers with a wide elevational distribution is the Hawaii Amakihi (Hernignathus virens). We measured morbidity and mortality in experimentally-infected Hawaii Amakihi that were captured in a high elevation, xeric habitat that is above the current range of the mosquito vector. Mortality among amakihi exposed to a single infective mosquito bite was 65% (13/20). All infected birds had significant declines in food consumption and a corresponding loss in body weight over the 60 day course of the experiment. Gross and microscopic lesions in birds that succumbed to malaria included enlargement and discoloration of the spleen and liver and parasitemias as high as 50% of circulating erythrocytes. Mortality in experimentally-infected amakihi was similar to that observed in Apapane (Himnatione sanguinea) and lower than that observed in Iiwi (Vestiaria coccinea) infected under similar conditions with the same parasite isolate. We conclude that the current elevational and geographic distribution of Hawaiian honeycreepers is determined by relative susceptibility to avian malaria.     

The effect of isolation,fragmentation, and population bottlenecks on song structure of a Hawaiian honeycreeper

Little is known about how important social behaviors such as song vary within and among populations for any of the endemic Hawaiian honeycreepers. Habitat loss and non‐native diseases (e.g., avian malaria) have resulted in isolation and fragmentation of Hawaiian honeycreepers within primarily high elevation forests. In this study, we examined how isolation of Hawai'i ‘amakihi (Chlorodrepanis virens) populations within a fragmented landscape influences acoustic variability in song. In the last decade, small, isolated populations of disease tolerant ‘amakihi have been found within low elevation forests, allowing us to record ‘amakihi songs across a large elevational gradient (10–1800 m) that parallels disease susceptibility on Hawai'i island. To understand underlying differences among populations, we examined the role of geographic distance, elevation, and habitat structure on acoustic characteristics of ‘amakihi songs. We found that the acoustic characteristics of ‘amakihi songs and song‐type repertoires varied most strongly across an elevational gradient. Differences in ‘amakihi song types were primarily driven by less complex songs (e.g., fewer frequency changes, shorter songs) of individuals recorded at low elevation sites compared to mid and high elevation populations. The reduced complexity of ‘amakihi songs at low elevation sites is most likely shaped by the effects of habitat fragmentation and a disease‐driven population bottleneck associated with avian malaria, and maintained through isolation, localized song learning and sharing, and cultural drift. These results highlight how a non‐native disease through its influence on population demographics may have also indirectly played a role in shaping the acoustic characteristics of a species.     

Experimental Evidence for Evolved Tolerance to Avian Malaria in a Wild Population of Low Elevation Hawai‘i ‘Amakihi (Hemignathus virens)

Introduced vector-borne diseases, particularly avian malaria (Plasmodium relictum) and avian pox virus (Avipoxvirus spp.), continue to play significant roles in the decline and extinction of native forest birds in the Hawaiian Islands. Hawaiian honeycreepers are particularly susceptible to avian malaria and have survived into this century largely because of persistence of high elevation refugia on Kaua‘i, Maui, and Hawai‘i Islands, where transmission is limited by cool temperatures. The long term stability of these refugia is increasingly threatened by warming trends associated with global climate change. Since cost effective and practical methods of vector control in many of these remote, rugged areas are lacking, adaptation through processes of natural selection may be the best long-term hope for recovery of many of these species. We document emergence of tolerance rather than resistance to avian malaria in a recent, rapidly expanding low elevation population of Hawai‘i ‘Amakihi (Hemignathus virens) on the island of Hawai‘i. Experimentally infected low elevation birds had lower mortality, lower reticulocyte counts during recovery from acute infection, lower weight loss, and no declines in food consumption relative to experimentally infected high elevation Hawai‘i ‘Amakihi in spite of similar intensities of infection. Emergence of this population provides an exceptional opportunity for determining physiological mechanisms and genetic markers associated with malaria tolerance that can be used to evaluate whether other, more threatened species have the capacity to adapt to this disease.     

Modeling Future Conservation of Hawaiian Honeycreepers by Mosquito Management and Translocation of Disease-Tolerant Amakihi

Avian malaria is an important cause of the decline of endemic Hawaiian honeycreepers. Because of the complexity of this disease system we used a computer model of avian malaria in forest birds to evaluate how two proposed conservation strategies: 1) reduction of habitat for mosquito larvae and 2) establishment of a low-elevation, malaria-tolerant honeycreeper (Hawaii Amakihi) to mid-elevation forests would affect native Hawaiian honeycreeper populations. We evaluated these approaches in mid-elevation forests, where malaria transmission is seasonal and control strategies are more likely to work. Our model suggests the potential benefit of larval habitat reduction depends on the level of malaria transmission, abundance of larval cavities, and the ability to substantially reduce these cavities. Permanent reduction in larval habitat of >80% may be needed to control abundance of infectious mosquitoes and benefit bird populations. Establishment of malaria-tolerant Amakihi in mid-elevation forests increases Amakihi abundance, creates a larger disease reservoir, and increases the abundance of infectious mosquitoes which may negatively impact other honeycreepers. For mid-elevation sites where bird populations are severely affected by avian malaria, malaria-tolerant Amakihi had little impact on other honeycreepers. Both management strategies may benefit native Hawaiian honeycreepers, but benefits depend on specific forest characteristics, the amount of reduction in larval habitat that can be achieved, and how malaria transmission is affected by temperature.     

Avian malaria Plasmodium relictum in native Hawaiian forest birds: epizootiology and demographic impacts on ‵apapane Himatione sanguinea

The role of introduced avian malaria Plasmodium relictum in the decline and extinction of native Hawaiian forest birds has become a classic example of the potential effect of invasive diseases on biological diversity of naïve populations. However, empirical evidence describing the impact of avian malaria on fitness of Hawai‵i's endemic forest birds is limited, making it difficult to determine the importance of disease among the suite of potential limiting factors affecting the distribution and abundance of this threatened avifauna. We combined epidemiological force‐of‐infection with multistate capture––recapture models to evaluate a 7‐year longitudinal study of avian malaria in ‵apapane, a relatively common native honeycreeper within mid‐elevation Hawaiian forests. We found that malaria transmission was seasonal in this mid‐elevation forest; transmission peaked during fall and during some years produced epizootic mortality events. Estimated annual mortality of hatch‐year birds typically exceeded 50% and mortality of adults exceeded 25% during epizootics. The substantial impact of avian malaria on this relatively common native species demonstrates the key role this disease has played in the decline and extinction of Hawaiian forest birds.     

Genetic structure and evolved malaria resistance in Hawaiian honeycreepers

Infectious diseases now threaten wildlife populations worldwide but population recovery following local extinction has rarely been observed. In such a case, do resistant individuals recolonize from a central remnant population, or do they spread from small, perhaps overlooked, populations of resistant individuals? Introduced avian malaria (Plasmodium relictum) has devastated low‐elevation populations of native birds in Hawaii, but at least one species (Hawaii amakihi, Hemignathus virens) that was greatly reduced at elevations below about 1000 m tolerates malaria and has initiated a remarkable and rapid recovery. We assessed mitochondrial and nuclear DNA markers from amakihi and two other Hawaiian honeycreepers, apapane (Himatione sanguinea) and iiwi (Vestiaria coccinea), at nine primary study sites from 2001 to 2003 to determine the source of re‐establishing birds. In addition, we obtained sequences from tissue from amakihi museum study skins (1898 and 1948–49) to assess temporal changes in allele distributions. We found that amakihi in lowland areas are, and have historically been, differentiated from birds at high elevations and had unique alleles retained through time; that is, their genetic signature was not a subset of the genetic variation at higher elevations. We suggest that high disease pressure rapidly selected for resistance to malaria at low elevation, leaving small pockets of resistant birds, and this resistance spread outward from the scattered remnant populations. Low‐elevation amakihi are currently isolated from higher elevations (> 1000 m) where disease emergence and transmission rates appear to vary seasonally and annually. In contrast to results from amakihi, no genetic differentiation between elevations was found in apapane and iiwi, indicating that slight variation in genetic or life‐history attributes can determine disease resistance and population recovery. Determining the conditions that allow for the development of resistance to disease is essential to understanding how species evolve resistance across a landscape of varying disease pressures.     

Detection of avian malaria (Plasmodium spp.) in native land birds of American Samoa

This study documents the presence ofPlasmodium spp. in landbirds ofcentral Polynesia. Blood samples collectedfrom eight native and introduced species fromthe island of Tutuila, American Samoa wereevaluated for the presence of Plasmodiumspp. by nested rDNA PCR, serology and/ormicroscopy. A total of 111/188 birds (59%)screened by nested PCR were positive. Detection of Plasmodium spp. was verifiedby nucleotide sequence comparisons of partial18S ribosomal RNA and TRAP(thrombospondin-related anonymous protein)genes using phylogenetic analyses. All samplesscreened by immunoblot to detect antibodiesthat cross-react with Hawaiian isolates of Plasmodium relictum (153) were negative. Lack of cross-reactivity is probably due toantigenic differences between the Hawaiian andSamoan Plasmodium isolates. Similarly,all samples examined by microscopy (214) werenegative. The fact that malaria is present,but not detectable by blood smear evaluation isconsistent with low peripheral parasitemiacharacteristic of chronic infections. Highprevalence of apparently chronic infections,the relative stability of the native land birdcommunities, and the presence of mosquitovectors which are considered endemic andcapable of transmitting avian Plasmodia,suggest that these parasites are indigenous toSamoa and have a long coevolutionary historywith their hosts.     

Changing climate and the altitudinal range of avian malaria in the Hawaiian Islands – an ongoing conservation crisis on the island of Kaua'i

Transmission of avian malaria in the Hawaiian Islands varies across altitudinal gradients and is greatest at elevations below 1500 m where both temperature and moisture are favorable for the sole mosquito vector, Culex quinquefasciatus, and extrinsic sporogonic development of the parasite, Plasmodium relictum. Potential consequences of global warming on this system have been recognized for over a decade with concerns that increases in mean temperatures could lead to expansion of malaria into habitats where cool temperatures currently limit transmission to highly susceptible endemic forest birds. Recent declines in two endangered species on the island of Kaua'i, the ‘Akikiki (Oreomystis bairdi) and ‘Akeke'e (Loxops caeruleirostris), and retreat of more common native honeycreepers to the last remaining high elevation habitat on the Alaka'i Plateau suggest that predicted changes in disease transmission may be occurring. We compared prevalence of malarial infections in forest birds that were sampled at three locations on the Plateau during 1994–1997 and again during 2007–2013, and also evaluated changes in the occurrence of mosquito larvae in available aquatic habitats during the same time periods. Prevalence of infection increased significantly at the lower (1100 m, 10.3% to 28.2%), middle (1250 m, 8.4% to 12.2%), and upper ends of the Plateau (1350 m, 2.0% to 19.3%). A concurrent increase in detections of Culex larvae in aquatic habitats associated with stream margins indicates that populations of the vector are also increasing. These increases are at least in part due to local transmission because overall prevalence in Kaua'i ‘Elepaio (Chasiempis sclateri), a sedentary native species, has increased from 17.2% to 27.0%. Increasing mean air temperatures, declining precipitation, and changes in streamflow that have taken place over the past 20 years are creating environmental conditions throughout major portions of the Alaka'i Plateau that support increased transmission of avian malaria.     

Avian Plasmodium infection in field‐collected mosquitoes during 2012–2013 in Tarlac,Philippines

Global warming threatens to increase the spread and prevalence of mosquito‐transmitted diseases. Certain pathogens may be carried by migratory birds and transmitted to local mosquito populations. Mosquitoes were collected in the northern Philippines during bird migration seasons to detect avian malaria parasites as well as for the identification of potential vector species and the estimation of infections among local mosquito populations. We used the nested PCR to detect the avian malaria species. Culex vishnui (47.6%) was the most abundant species collected and Cx. tritaeniorhynchus (13.8%) was the second most abundant. Avian Plasmodium parasites were found in eight mosquito species, for which the infection rates were between 0.5% and 6.2%. The six Plasmodium genetic lineages found in this study included P. juxtanucleare ‐GALLUS02, Tacy7 (Donana04), CXBIT01, Plasmodium species LIN2 New Zealand, and two unclassified lineages. The potential mosquito vectors for avian Plasmodium parasites in the Philippines were Cq. crassipes, Cx. fuscocephala, Cx. quinquefasciatus, Cx. sitiens, Cx. vishnui, and Ma. Uniformis; two major genetic lineages, P. juxtanucleare and Tacy7, were identified.     

Climate change increases the risk of malaria in birds

Malaria caused by Plasmodium parasites is one of the worst scourges of mankind and threatens wild animal populations. Therefore, identifying mechanisms that mediate the spread of the disease is crucial for both human health and conservation. Human‐induced climate change has been hypothesized to alter the geographic distribution of malaria pathogens. As the earth warms, arthropod vectors may display a general range expansion or may enjoy longer breeding season, both of which can enhance parasite transmission. Moreover, Plasmodium species may directly benefit for elevating temperatures, which provide stimulating conditions for parasite reproduction. To test for the link between climate change and malaria prevalence on a global scale for the first time, I used long‐term records on avian malaria, which is a key model for studying the dynamics of naturally occurring malarial infections. Following the variation in parasite prevalence in more than 3000 bird species over seven decades, I show that the infection rate by Plasmodium is strongly associated with temperature anomalies and has been augmented with accelerating tendency during the last 20 years. The impact of climate change on malaria prevalence varies across continents, with the strongest effects found for Europe and Africa. Migration habit did not predict susceptibility to the escalating parasite pressure by Plasmodium. Consequently, wild birds are at an increasing risk of malaria infection due to recent climate change, which can endanger both naïve bird populations and domesticated animals. The prevailing avian example may provide useful lessons for understanding the effect of climate change on malaria in humans.     

Will a warmer and wetter future cause extinction of native Hawaiian forest birds?

Isolation of the Hawaiian archipelago produced a highly endemic and unique avifauna. Avian malaria (Plasmodium relictum), an introduced mosquito‐borne pathogen, is a primary cause of extinctions and declines of these endemic honeycreepers. Our research assesses how global climate change will affect future malaria risk and native bird populations. We used an epidemiological model to evaluate future bird–mosquito–malaria dynamics in response to alternative climate projections from the Coupled Model Intercomparison Project. Climate changes during the second half of the century accelerate malaria transmission and cause a dramatic decline in bird abundance. Different temperature and precipitation patterns produce divergent trajectories where native birds persist with low malaria infection under a warmer and dryer projection (RCP4.5), but suffer high malaria infection and severe reductions under hot and dry (RCP8.5) or warm and wet (A1B) futures. We conclude that future global climate change will cause significant decreases in the abundance and diversity of remaining Hawaiian bird communities. Because these effects appear unlikely before mid‐century, natural resource managers have time to implement conservation strategies to protect this unique avifauna from further decimation. Similar climatic drivers for avian and human malaria suggest that mitigation strategies for Hawai'i have broad application to human health.     

Is Avian Malaria Playing a Role in Native Bird Declines in New Zealand? Testing Hypotheses along an Elevational Gradient

The mosquito-borne disease avian malaria (Plasmodium spp.) has impacted both captive populations and wild individuals of native New Zealand bird species. However, whether or not it is a cause of concern to their wild populations is still unclear. In Hawaii, the disease has been a major factor in the population declines of some native forest bird species, often limiting their elevational distribution due to an inverse relationship between force of infection and elevation. While studies have investigated latitudinal patterns of infection in New Zealand, elevational patterns are unexplored. To address this, a survey was conducted in Nelson Lakes National Park, a site experiencing native bird declines in which disease has been suggested as playing a role, to investigate whether there is a similar inverse relationship in New Zealand. Results from blood samples (n = 436) collected over three seasons across a broad elevational range (650–1400 m) support there being such a relationship. In addition, an overall higher prevalence in non-native (14.1%) versus native birds (1.7%) may indicate differential impacts on these two groups, while particularly high prevalence in non-native Turdus spp. supports previous suggestions that they are key reservoir hosts for the disease. Overall, these findings add weight to the hypothesis that avian malaria is playing a role in ongoing declines of native New Zealand birds.     

Seasonal dynamics in mosquito abundance and temperature do not influence avian malaria prevalence in the Himalayan foothills

We examined seasonal prevalence in avian haemosporidians (Plasmodium and Haemoproteus) in migrant and resident birds in western Himalaya, India. We investigated how infection with haemosporidians in avian hosts is associated with temporal changes in temperature and mosquito abundance along with host abundance and life‐history traits (body mass). Using molecular methods for parasite detection and sequencing partial cytochrome b gene, 12 Plasmodium and 27 Haemoproteus lineages were isolated. Our 1‐year study from December 2008 to December 2009 in tropical Himalayan foothills revealed a lack of seasonal variation in Plasmodium spp. prevalence in birds despite a strong correlation between mosquito abundance and temperature. The probability of infection with Plasmodium decreased with increase in temperature. Total parasite prevalence and specifically Plasmodium prevalence showed an increase with average avian body mass. In addition, total prevalence exhibited a U‐shaped relationship with avian host abundance. There was no difference in prevalence of Plasmodium spp. or Haemoproteus spp. across altitudes; parasite prevalence in high‐altitude locations was mainly driven by the seasonal migrants. One Haemoproteus lineage showed cross‐species infections between migrant and resident birds. This is the first molecular study in the tropical Himalayan bird community that emphasizes the importance of studying seasonal variation in parasite prevalence. Our study provides a basis for further evolutionary study on the epidemiology of avian malaria and spread of disease across Himalayan bird communities, which may not have been exposed to vectors and parasites throughout the year, with consequential implications to the risk of infection to naïve resident birds in high altitude.     

Implications of Plasmodium parasite infected mosquitoes on an insular avifauna: the case of Socorro Island,México

Avian malaria (Plasmodium spp.) has been implicated in the decline of avian populations in the Hawaiian Islands and it is generally agreed that geographically isolated and immunologically naïve bird populations are particularly vulnerable to the pathogenic effects of invasive malaria parasites. In order to assess the potential disease risk of malaria to the avifauna of Socorro Island, México, we surveyed for Plasmodium isolates from 1,300 resident field‐caught mosquitoes. Most of them were identified as Aedes (Ochlerotatus) taeniorhynchus (Wiedemann, 1821), which were abundant in the salt marshes. We also collected Culex quinquefasciatus Say, 1823 close to human dwellings. Mitochondrial ND5 and COII gene sequences of Ae. taeniorhynchus were analyzed and compared to corresponding sequences of mosquitoes of the Galápagos Islands, Latin America, and the North American mainland. Aedes lineages from Socorro Island clustered most closely with a lineage from the continental U.S. Plasmodium spp. DNA was isolated from both species of mosquitoes. From 38 positive pools, we isolated 11 distinct mitochondrial Cytb lineages of Plasmodium spp. Seven of the Plasmodium lineages represent previously documented avian infective strains while four were new lineages. Our results confirm a potential risk for the spread of avian malaria and underscore the need to monitor both the mosquito and avian populations as a necessary conservation measure to protect endangered bird species on Socorro Island.     

Artificial incubation,hand-rearing,behavior, and release of common 'Amakihi (Hemignathus virens virens): Surrogate research for restoration of endangered Hawaiian forest birds

In order to test the effectiveness of captive-rearing and release strategies for future restoration of birds in Hawai'i, this pilot study was conducted in forests where introduced avian disease and mammalian predators were present. Methodology used resulted in the first successful hatching of Drepanidinae eggs in an incubator and subsequent hand-rearing of chicks from hatch. Sixteen Common 'Amakihi (Hemignathus virens virens) (mean hatch weight = 1.4 g) were hand-reared. Two different reintroduction strategies were evaluated for small honeycreepers. Known mortality in the wild after release was due to mosquito-transmitted disease (avian malaria and pox). This pilot study shows that the techniques necessary to hatch, rear, and release endangered Hawaiian honeycreepers are available. However, restoration efforts will probably not succeed unless mosquito-free, predator-controlled reintroduction sites are available or strategies are developed to decrease mortality in naive honeycreepers exposed to disease after release. © 1996 Wiley-Liss, Inc.     

Natural selection of the major histocompatibility complex (Mhc) in Hawaiian honeycreepers (Drepanidinae)

The native Hawaiian honeycreepers represent a classic example of adaptive radiation and speciation, but currently face one the highest extinction rates in the world. Although multiple factors have likely influenced the fate of Hawaiian birds, the relatively recent introduction of avian malaria is thought to be a major factor limiting honeycreeper distribution and abundance. We have initiated genetic analyses of class II beta chain Mhc genes in four species of honeycreepers using methods that eliminate the possibility of sequencing mosaic variants formed by cloning heteroduplexed polymerase chain reaction products. Phylogenetic analyses group the honeycreeper Mhc sequences into two distinct clusters. Variation within one cluster is high, with dN > dS and levels of diversity similar to other studies of Mhc (B system) genes in birds. The second cluster is nearly invariant and includes sequences from honeycreepers (Fringillidae), a sparrow (Emberizidae) and a blackbird (Emberizidae). This highly conserved cluster appears reminiscent of the independently segregating Rfp-Y system of genes defined in chickens. The notion that balancing selection operates at the Mhc in the honeycreepers is supported by transpecies polymorphism and strikingly high dN/dS ratios at codons putatively involved in peptide interaction. Mitochondrial DNA control region sequences were invariant in the i'iwi, but were highly variable in the 'amakihi. By contrast, levels of variability of class II beta chain Mhc sequence codons that are hypothesized to be directly involved in peptide interactions appear comparable between i'iwi and 'amakihi. In the i'iwi, natural selection may have maintained variation within the Mhc, even in the face of what appears to a genetic bottleneck.     

High prevalence of haemosporidians in Reed Warbler Acrocephalus scirpaceus and Sedge Warbler Acrocephalus schoenobaenus in Spain

Apicomplexan blood parasites (genera Haemoproteus, Plasmodium and Leucocytozoon) prevalence in two related species (Reed Warbler Acrocephalus scirpaceus and Sedge Warbler A. schoenobaenus) was studied in 2006 at the Natural Reserve of Castronu?o-Vega del Duero, Western Spain, a stopover area during the autumn migration. A fragment of the mitochondrial cytochrome b gene of the parasites was amplified, using a nested PCR assay, from avian blood samples. High prevalence of malaria parasites was found in both species, 84.6% in Reed Warbler and 71.8% in Sedge Warbler, and the degree of infection reach 100% of the population that breed at the Reserve, suggesting good conditions for the development of dipteran vectors in this area. By sequencing 464 nucleotides of the obtained fragments, we found four different mitochondrial haplotypes of Haemoproteus or Plasmodium in the two species analysed. Leucocytozoon infection was not detected, in contrast to the high prevalence of this parasite in other avian species in Spain, probably because the water course studied is not an adequate habitat for its vectors.     

Parallel evolution of gene classes,but not genes: Evidence from Hawai'ian honeycreeper populations exposed to avian malaria

Adaptation in nature is ubiquitous, yet characterizing its genomic basis is difficult because population demographics cause correlations with nonadaptive loci. Introduction events provide opportunities to observe adaptation over known spatial and temporal scales, facilitating the identification of genes involved in adaptation. The pathogen causing avian malaria, Plasmodium relictum, was introduced to Hawai'i in the 1930s and elicited extinctions and precipitous population declines in native honeycreepers. After a sharp initial population decline, the Hawai'i ‘amakihi (Chlorodrepanis virens) has evolved tolerance to the parasite at low elevations where P. relictum exists, and can sustain infection without major fitness consequences. High‐elevation, unexposed populations of ‘amakihi display little to no tolerance. To explore the genomic basis of adaptation to P. relictum in low‐elevation ‘amakihi, we genotyped 125 ‘amakihi from the island of Hawai'i via hybridization capture to 40,000 oligonucleotide baits containing SNPs and used the reference ‘amakihi genome to identify genes potentially under selection from malaria. We tested for outlier loci between low‐ and high‐elevation population pairs and identified loci with signatures of selection within low‐elevation populations. In some cases, genes commonly involved in the immune response (e.g., major histocompatibility complex) were associated with malaria presence in the population. We also detected several novel candidate loci that may be implicated in surviving malaria infection (e.g., beta‐defensin, glycoproteins and interleukin‐related genes). Our results suggest that rapid adaptation to pathogens may occur through changes in different immune genes, but in the same classes of genes, across populations.     

Diversity,origins and virulence of <Emphasis Type="Italic">Avipoxviruses</Emphasis> in Hawaiian Forest Birds

We cultured avian pox (Avipoxvirus spp.) from lesions collected on Hawai‘i, Maui, Moloka‘i, and ‘Oahu in the Hawaiian Islands from 15 native or non-native birds representing three avian orders. Phylogenetic analysis of a 538 bp fragment of the gene encoding the virus 4b core polypeptide revealed two distinct variant clusters, with sequences from chickens (fowlpox) forming a third distinct basal cluster. Pox isolates from one of these two clusters appear closely related to canarypox and other passerine pox viruses, while the second appears more specific to Hawai‘i. There was no evidence that birds were infected simultaneously with multiple pox virus variants based on evaluation of multiples clones from four individuals. No obvious temporal or geographic associations were observed and strict host specificity was not apparent among the 4b-defined field isolates. We amplified a 116 bp 4b core protein gene fragment from an ‘Elepaio (Chasiempis sandwichensis) collected in 1900 on Hawai‘i Island that clustered closely with the second of the two variants, suggesting that this variant has been in Hawai‘i for at least 100 years. The high variation detected between the three 4b clusters provides evidence for multiple, likely independent introductions, and does not support the hypothesis of infection of native species through introduction of infected fowl. Preliminary experimental infections in native Hawai‘i ‘Amakihi (Hemignathus virens) suggest that the 4b-defined variants may be biologically distinct, with one variant appearing more virulent. These pox viruses may interact with avian malaria (Plasmodium relictum), another introduced pathogen in Hawaiian forest bird populations, through modulation of host immune responses.     

Loss of forest cover and host functional diversity increases prevalence of avian malaria parasites in the Atlantic Forest

Host phylogenetic relatedness and ecological similarity are thought to contribute to parasite community assembly and infection rates. However, recent landscape level anthropogenic changes may disrupt host-parasite systems by impacting functional and phylogenetic diversity of host communities. We examined whether changes in host functional and phylogenetic diversity, forest cover, and minimum temperature influence the prevalence, diversity, and distributions of avian haemosporidian parasites (genera Haemoproteus and Plasmodium) across 18 avian communities in the Atlantic Forest. To explore spatial patterns in avian haemosporidian prevalence and taxonomic and phylogenetic diversity, we surveyed 2241 individuals belonging to 233 avian species across a deforestation gradient. Mean prevalence and parasite diversity varied considerably across avian communities and parasites responded differently to host attributes and anthropogenic changes. Avian malaria prevalence (termed herein as an infection caused by Plasmodium parasites) was higher in deforested sites, and both Plasmodium prevalence and taxonomic diversity were negatively related to host functional diversity. Increased diversity of avian hosts increased local taxonomic diversity of Plasmodium lineages but decreased phylogenetic diversity of this parasite genus. Temperature and host phylogenetic diversity did not influence prevalence and diversity of haemosporidian parasites. Variation in the diversity of avian host traits that promote parasite encounter and vector exposure (host functional diversity) partially explained the variation in avian malaria prevalence and diversity. Recent anthropogenic landscape transformation (reduced proportion of native forest cover) had a major influence on avian malaria occurrence across the Atlantic Forest. This suggests that, for Plasmodium, host phylogenetic diversity was not a biotic filter to parasite transmission as prevalence was largely explained by host ecological attributes and recent anthropogenic factors. Our results demonstrate that, similar to human malaria and other vector-transmitted pathogens, prevalence of avian malaria parasites will likely increase with deforestation.