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Understanding factors that influence the spatial and temporal distributions of blood parasites is important to help predict how host species and their parasites may respond to global change. Factors that may influence parasite distributions are land cover and host dispersal patterns, which may result in exposure of a host to novel parasites, or escape from parasites of their origin. We screened golden-winged warblers from across the United States and Canada for blood parasites, and investigated whether land-use patterns or host dispersal affected the prevalence and composition of haemosporidian assemblages. Parasite prevalence varied strongly with study area, and areas with high agricultural cover had a significantly higher prevalence of Leucocytozoon and Parahaemoproteus parasites. Lineages of Parahaemoproteus and Leucocytozoon were genetically differentiated among study areas, and prevalence and composition of parasite assemblages indicated an increase in parasite prevalence and accumulation of unique parasite lineages from the southeast to the northwest. This matches the historical range expansion and natal dispersal patterns of golden-winged warblers, and suggests that golden-winged warblers may have been sensitive to novel parasites as they dispersed. The high prevalence and diversity of parasite lineages in the north-west extent of their breeding range (Manitoba) indicates that this population may face unique pressures.

SUMMARYThe population growth of endangered whooping cranes (Grus americana) is not consistent with species recovery goals, and the impact of parasite infection on whooping crane populations is largely unknown. Disease ecology and epidemiology research of endangered species is often hindered by limited ability to conduct invasive sampling on the target taxa. Accordingly, we hypothesized that sandhill cranes (Grus canadensis) would be a useful surrogate species to investigate the health impacts of Haemosporida infection in whooping cranes. Our goal was to compare the prevalence and diversity of Haemosporida infection between whooping cranes and sandhill cranes. We detected an overall infection prevalence of 83·6% (n = 61) in whooping cranes and 59·6% (n = 47) and 63·6 (n = 22) in two sympatric sandhill crane populations captured in Texas. Prevalence was significantly lower in allopatric sandhill cranes captured in New Mexico (12·1%, n = 33). Haemoproteus antigonis was the most abundant haemoparasite in cranes, present in 57·4% of whooping cranes and 39·2% of sandhill cranes; Plasmodium and Leucocytozoon were present at significantly lower levels. The high prevalence of Haemosporida in whooping cranes and sympatric sandhill cranes, with shared parasite lineages between the two species, supports sandhill cranes as a surrogate species for understanding health threats to endangered whooping cranes.

SUMMARYThe diversity of avian malaria parasites is much greater than 20th century morphologists realized and virtually every study in this field in the last 15 years has uncovered previously undocumented diversity at multiple levels within the taxonomic hierarchy. Despite this explosion of knowledge, there remain vast sampling gaps, both geographically and host-taxonomically, which makes characterizing patterns of diversity extremely challenging. Here, we summarize the current state of knowledge of sub-Saharan African avian malaria parasite diversity, focusing on avian hosts endemic to Africa. The relative proportions of the parasite genera included here, Plasmodium, Haemoproteus (including Parahaemoproteus) and Leucocytozoon, varied between regions, in part due to habitat preferences of the insect vectors of these genera, and in part we believe due to sampling bias. Biogeographic regions of sub-Saharan Africa harbour about the same proportion of endemic to shared parasite lineages, but there appears to be no phylogenetic structuring across regions. Our results highlight the sampling problem that must be addressed if we are to have a detailed understanding of parasite diversity in Africa. Without broad sampling within and across regions and hosts, using both molecular tools and microscopy, conclusions about parasite diversity, host–parasite interactions or even transmission dynamics remain extremely limited.

The biogeographic histories of parasites and pathogens are infrequently compared with those of free-living species, including their hosts. Documenting the frequency with which parasites and pathogens disperse across geographic regions contributes to understanding not only their evolution, but also the likelihood that they may become emerging infectious diseases. Haemosporidian parasites of birds (parasite generaPlasmodium,HaemoproteusandLeucocytozoon) are globally distributed, dipteran-vectored parasites. To date, over 2000 avian haemosporidian lineages have been designated by molecular barcoding methods. To achieve their current distributions, some lineages must have dispersed long distances, often over water. Here we quantify such events using the global avian haemosporidian database MalAvi and additional records primarily from the Americas. We scored lineages as belonging to one or more global biogeographic regions based on infection records. Most lineages were restricted to a single region but some were globally distributed. We also used part of the cytochromebgene to create genus-level parasite phylogenies and scored well-supported nodes as having descendant lineages in regional sympatry or allopatry. Descendant sister lineages ofPlasmodium,HaemoproteusandLeucocytozoonwere distributed in allopatry in 11, 16 and 15% of investigated nodes, respectively. Although a small but significant fraction of the molecular variance in cytochromebof all three genera could be explained by biogeographic region, global parasite dispersal likely contributed to the majority of the unexplained variance. Our results suggest that avian haemosporidian parasites have faced few geographic barriers to dispersal over their evolutionary history.

SUMMARYSeabirds are often free from blood parasites, and a recent review suggested that phylogenetic, ecological and life-history parameters can determine the prevalence of blood parasites in seabirds. However, there is a lack of data available from many seabird groups, and a larger database is needed to understand prevalence patterns of blood parasites. We used a molecular screening approach to detect parasites of the genera Plasmodium, Haemoproteus, Leucocytozoon and Babesia in five species of two genera of seabirds that breed on Atlantic Ocean islands off Brazil. The observed patterns differed between the two bird genera. Like other Laridae, brown noddy, Anous stolidus adults were infected with Haemoproteus with low prevalence. Masked boobies, Sula dactylatra and brown boobies, Sula leucogaster were infected with Babesia. Of the latter, mainly juveniles were infected. In all species, intensity of infection (i.e. number of infected erythrocytes) was so low that parasites remained undetected in blood smears. This may explain the absence of major effects on the body condition of birds, although infected juvenile masked boobies were lighter than juveniles that were not infected with Babesia. Two tree-nesting species; black noddy, Anous minutus and red-footed booby, Sula sula did not have blood parasites, suggesting that tree-nesting may reduce the exposure to arthropod vectors compared with ground nesting in these species.

The group of haemosporidian parasites is of general interest to basic and applied science, since several species infect mammals, leading to malaria and associated disease symptoms. Although the great majority of haemosporidian parasites appear in bird hosts, as in the case of Leucocytozoon buteonis, there is little genomic information about genetic aspects of their co-evolution with hosts. Consequently, there is a high need for parasite-enrichment strategies enabling further analyses of the genomes, namely without exposure to DNA-intercalating dyes. Here, we used flow cytometry without an additional labelling step to enrich L. buteonis from infected buzzard blood. A specific, defined area of two-dimensional scattergramms was sorted and the fraction was further analysed. The successful enrichment of L. buteonis in the sorted fraction was demonstrated by Giemsa-staining and qPCR revealing a clear increase of parasite-specific genes, while host-specific genes were significantly decreased. This is the first report describing a labelling-free enrichment approach of L. buteonis from infected buzzard blood. The enrichment of parasites presented here is free of nucleic acid-intercalating dyes which may interfere with fluorescence-based methods or subsequent sequencing approaches.

SUMMARYWe measured seasonal changes in the prevalence of haematozoa (Leucocytozoon fringillinarum, Haemoproteus fringillae, andTrypanosoma avium) in free-ranging White-winged Crossbills,Loxia leucoptera, over 1·5 year in Fairbanks, Alaska, USA. This prevalence was low during early winter.L. fringillinarumprevalence increased in late winter/early spring, in the absence of vectors, suggesting relapse of latent infection. By contrast, the prevalence ofT. aviumandH. fringillaedid not increase until mid-spring, coincident with the emergence of putative vectors and suggestive of new inoculations. The winter breeding period was not associated with lower body condition or elevated blood heterophil/lymphocyte ratios than the summer post-breeding period. Thus, birds unlikely perceived their breeding effort as particularly stressful. Adult males in May and June had low plasma testosterone and their blood prevalence ofL. fringillinarum, but not other haemoparasites, was higher than in adult females. This difference may have resulted from sex differences in behaviour and/or plumage colouration – bright red in males, dull green/yellow in females. Species in which reproduction and vector abundance are seasonally dissociated may constitute important models for investigating the respective contribution of reproductive hormones, breeding effort, and vector abundance to patent and latent hemoparasitic infections and to new inoculations.

More than 200 species of avian Haemosporidia (genera Plasmodium, Haemoproteus, and Leucocytozoon) have been described based primarily on morphological characters seen in blood smears. Recent molecular studies, however, suggest that such methods may mask a substantial cryptic diversity of avian haemosporidians. We surveyed the haemosporidians of birds sampled at 1 site in Israel. Parasites were identified to species based on morphology, and a segment of the parasite's cytochrome b gene was sequenced. We compared 3 species concepts: morphological, genetic, and phylogenetic. Fifteen morphological species were present. Morphological species that occurred once within our dataset were associated with a unique gene sequence, displayed large genetic divergence from other morphological species, and were not contained within clades of morphological species that occurred more than once. With only 1 exception, morphological species that were identified from multiple bird hosts presented identical sequences for all infections, or differed by few synonymous substitutions, and were monophyletic for all phylogenetic analyses. Only the morphological species Haemoproteus belopolskyi did not follow this trend, falling instead into at least 2 genetically distant clades. Thus, except for H. belopolskyi, parasites identified to species by morphology were supported by both the genetic and phylogenetic species concepts.

SUMMARYYellow-eyed penguins (YEPs) have suffered major population declines over the past 30 years, with no single cause established.Leucocytozoonwas first identified in yellow-eyed penguins in 2005. During the 2008/09 breeding season, a high mortality was seen in both mainland yellow-eyed penguins as well as those on Enderby Island of the Auckland Islands archipelago. A high overall prevalence ofLeucocytozoonspp. in association with a high incidence of chick mortality was observed during this period on Enderby Island. One chick had histological evidence of leucocytozoonosis with megaloschizonts in multiple organs throughout its body. In addition, a high prevalence (73·7%) ofLeucocytozoonwas observed by PCR in the blood of adult Enderby yellow-eyed penguins taken during the 2006/07 season. These findings were different from the low prevalence detected by PCR on the coast of the South Island (11%) during the 2008/2009 breeding session and earlier on Campbell Island (21%) during the 2006/2007 breeding session. TheLeucocytozoonspp. sequences detected lead us to conclude that theLeucocytozoonparasite is common in yellow-eyed penguins and has a higher prevalence in penguins from Enderby Island than those from Campbell Island and the mainland of New Zealand. The Enderby Island yellow-eyed penguins are infected with aLeucocytozoonspp. that is genetically distinct from that found in other yellow-eyed penguin populations. The role ofLeucocytozoonin the high levels of chick mortality in the yellow-eyed penguins remains unclear.

SUMMARYParasite prevalence is an important variable in many evolutionary and ecological studies. In birds, haemosporidian blood parasites have been in focus of many comparative analyses. Because low prevalence is difficult to estimate precisely and that studies finding low prevalence are more likely to remain unpublished, our knowledge of parasite prevalence is biased towards highly infected taxa. Species with naturally low levels of infection are nonetheless interesting as they may provide models for studying the evolution of pathogen resistance. In the present study we show that the prevalence of Haemoproteus parasites is markedly lower in several taxa within the widely distributed chiffchaff species-complex compared to other species within the genus Phylloscopus. Since chiffchaffs, P. collybita, commonly coexists in the same habitat as congeners frequently infected with Haemoproteus parasites, immediate ecological variables like abundance of vectors can hardly explain this difference. Some of the parasites infecting coexisting congeners are broad host generalists leaving it enigmatic why chiffchaffs are almost free of Haemoproteus infections. We propose that detailed infection experiments are needed to illuminate whether chiffchaffs possess a genetic immunity against Haemoproteus parasites or if other more subtle ecological processes, like anti-vector behaviour, play a role in its generally low level of infestation.

SUMMARYThe highly pathogenic avian protozoanLeucocytozoon caulleryiinfects host chicken cells, and interference by the host genome results in difficulty in obtaining protozoal DNA for genetic analysis. We used flow cytometry analysis to separate expelledL. caulleryigametocytes from infected chicken blood and to analyse cell populations and sorting by FACS efficiency. Infected blood cells stained with SYTO-24 showed a specific area on 2-dimensional scattergrams compared to uninfected blood. The specific area was sorted, and approximately 85% of the sorted cells were identified asL. caulleryigametocytes by microscopic observation. DNA was also extracted from the sorted fraction, and a clear increase in polymerase chain reaction (PCR) amplification of protozoal DNA was observed compared to infected blood without sorting. Host-derived DNA was also detected by PCR; however, its amplification was decreased compared to that in unsorted infected blood. This is the first report of the separation ofL. caulleryigametocytes from infected host blood using flow cytometry. This method may be applied to further genetic analyses such as studies of the dynamics of stage-specificL. caulleryigene expression.

SUMMARYYellow-eyed penguins on Stewart Island were identified with a Leucocytozoon spp. of a novel lineage in association with a high regional incidence of chick mortality (n=32, 100% mortality) during the November 2006 to January 2007 breeding season. Fourteen chicks from Stewart Island were examined post-mortem and histologically for Leucocytozoon infection. In addition, a survey of blood to detect Leucocytozoon spp. infections using PCR was performed on 107 yellow-eyed penguins from 4 distinct nesting areas on the South Island (Oamaru, Otago Peninsula, and Catlins) (n=95), and Stewart Island (n=12). The results of the study revealed that 2 of the 14 (14%) chicks necropsied showed severe, disseminated megaloschizont formation in the liver, spleen, lung, kidney and other tissues characteristic of leucocytozoonosis. Eighty-three percent (83%) of blood samples collected from Stewart Island penguins contained Leucocytozoon DNA, whereas samples from the 3 other nesting areas were negative for the blood parasite. Leucocytozoon spp. DNA sequences isolated from blood and tissues of adults (n=10) and chicks (n=7) were similar and grouped with other published Leucocytozoon spp. sequences but in a distinct cluster together with closely related isolates from a Western march harrier (Circus aerginosus) and common loon (Gavia immer). These findings suggest that yellow-eyed penguins on Stewart Island are infected with a regionally isolated, host-specific Leucocytozoon spp. which may contribute to the high chick mortality observed during this period.

In the last issue of Parasitology a description is given by Wenyon (5) of the developmental cycle of Leucocytozoon canis in the tick Rhipicephalus sanguineus. Wenyon refers to the description of the development given by me (2), but rightly points out that one stage, that of the large compound cysts, was overlooked. The surmise, however, that these large and easily ruptured cysts were overlooked owing to their having been broken up in the preparation of films is only part of the truth. A full explanation involves facts in regard to the development of this parasite which have not been touched upon by Wenyon; it is possible, as will be seen shortly, that Wenyon's description of the cycle, like mine, involved a considerable error. In this respect, as a result of work not yet published, I am able to supplement what has been written on the subject.

Of 63 specimens of the lizardMabuya mabouya(Scincidae) examined in Pará State, Brazil, 31 showed one or more haemosporidians in the peripheral blood. These includedGarnia(Plasmodium)morula(Telford, 1970), Lainson, Landau & Shaw, 1971;Plasmodium tropiduriandP. diploglossiAragão & Neiva, 1909; aPlasmodiumspecies close toP. diminutivumTelford, 1973a; and a new species ofSaurocytozoonLainson & Shaw, 1969a. Eleven different combinations of these parasites were noted, emphasizing the care needed in avoiding erroneous descriptions based on mixed infections. This is especially so whenG. morulais mixed withP. tropidurior other species with small, rounded gametocytes occupying a polar position in the red blood cells. Of 18 skinks infected withG. morula, 10 had pure infections, as judged by repeated examinations over long periods. The morphology of the Brazilian parasite is very similar to that described forG. morulafrom Panama (Telford, 1970): no malarial pigment could be demonstrated in any stage of the infection, including ookinetes and possible early oocysts which were studied in experimentally infectedCulex pipiens fatigans. Ookinetes and rounded forms, possibly representing uninucleate oocysts, persisted in these insects up to 24 h after the blood-meal, but developed no further.C. p. fatigansis thus unlikely to be a natural vector ofG. morula. Saurocytozoon mabuyisp.nov. is described inM. mabouya: the gametocytes develop in the lymphocytes and monocytes of the circulating blood and reach maximum size in 19–28 days. The parasite can be distinguished from the other known species,S. tupinambiof the teiid lizardTupinambus nigropunctatus, by its smaller gametocytes. Nothing is known of the sporogonic cycle. Differences between the sporogonic stages ofS. tupinambiandLeucocytozoonare listed.

Saurocytozoon tupinambi gen.nov., sp.nov. (Haemosporidiidea, Leucocytozoidae) is described in the teiid lizard, Tupinambus nigropunctatus, from Pará State, Brazil. This would appear to be the first record of a member of this family in a reptile; the existing genera Leucocytozoon and Akiba are known only in birds.The gametocytes of S. tupinambi develop within lymphocytes and immature erythrocytes. Only developing and mature gametocytes are found in the peripheral blood: morphologically they are very similar to the gametocytes of Leucocytozoon and Akiba. No mega-schizonts or other tissue stages have yet been detected.Aedes spp., have been trapped while feeding on Tupinambus in areas where infected lizards have been caught, and may possibly be the vectors.We are indebted to Professor P. C. C. Garnham, the London School of Hygiene and Tropical Medicine, London, who has examined our material and offered his invaluable opinions.The cost of the colour plate was defrayed by the Wellcome Parasitology Unit, Instituto Evandro Chagas, Fundaçao Serviço Especial de Saúde Pública, Belem, Pará, Brazil.