Angie D. González

Mixed species flock, nest height, and elevation partially explain avian haemoparasite prevalence in Colombia

The high avian biodiversity present in the Neotropical region offers a great opportunity to explore the ecology of host-parasite relationships. We present a survey of avian haemoparasites in a megadiverse country and explore how parasite prevalences are related to physical and ecological host characteristics. Using light microscopy, we documented the presence of haemoparasites in over 2000 individuals belonging to 246 species of wild birds, from nine localities and several ecosystems of Colombia. We analysed the prevalence of six avian haemoparasite taxa in relation to elevation and the following host traits: nest height, nest type, foraging strata, primary diet, sociality, migratory behaviour, and participation in mixed species flocks. Our analyses indicate significant associations between both mixed species flocks and nest height and Haemoproteus and Leucocytozoon prevalence. The prevalence of Leucocytozoon increased with elevation, whereas the prevalence of Trypanosoma and microfilariae decreased. Plasmodium and Haemoproteus prevalence did not vary significantly with elevation; in fact, both parasites were found up to 3300m above sea level. The distribution of parasite prevalence across the phylogeny of bird species included in this study showed little host phylogenetic signal indicating that infection rates in this system are evolutionarily labile. Vector distribution as well as the biology of transmission and the maintenance of populations of avian haemoparasites deserve more detailed study in this system.

Avian haemosporidians from Neotropical highlands: Evidence from morphological and molecular data.

Avian haemosporidian parasites have been scarcely studied in the Neotropical highlands despite the high avian diversity reported and the uniqueness of these ecosystems. The aims of this study were to examine Haemoproteus and Plasmodium diversity based on morphological and molecular data, as well as to explore the concordance between these two approaches, when identifying species. We sampled 1487 birds belonging to 166 species, in localities of the Colombian Andean region at elevations ranging from 2100 to 4000 m above sea level. Here, we report twelve morphological parasite species, of which five are undescribed. Thirty parasite cytochrome b lineages are reported, 17 of which for the first time. We provide morphological information and illustrations, as well as, cytochrome b lineages for six morphospecies: Haemoproteus columbae, Haemoproteus witti, Haemoproteus coatneyi, Haemoproteus vireonis, Plasmodium lutzi, and Plasmodium unalis. This is the first report to provide a linkage between morphology and a molecular lineage for H. witti. Cytochrome b gene proved to be useful for species determination as DNA barcoding. Differences in parasite composition between lowlands and highlands in Colombia suggest a replacement of avian Plasmodium fauna. Parasite lineages restricted to either Colombian resident or Nearctic migratory birds were found; but a single lineage common in both has not been recorded in Nearctic non-migratory birds. We generated valuable information by using both morphological and molecular data representing competent host-parasite relationships which are based on observation of gametocytes in circulation; and increased the taxon sampling of avian haemosporidian.

Identification of Plasmodium (Haemamoeba) lutzi (Lucena, 1939) from Turdus fuscater (Great Thrush) in Colombia

Abstract:  This study reports a broadening of the altitudinal range and a new host for Plasmodium (Haemamoeba) lutzi in Colombia. The study was conducted in the city of Bogotá, located in the Eastern Cordillera of Colombia at 2,560 m asl (meters above sea level) with an average annual temperature of 15 C. In total, 156 specimens of birds belonging to 25 species and 14 families were captured using mist nets. The blood samples were collected through venipuncture and analyzed by light microscopy. Plasmodium (H.) lutzi was only found in 2 individuals of Turdus fuscater (Great Thrush). This parasite has previously been reported in Aramides cajaneus (before: Aramides cajanea) (Grey-Necked Wood Rail), a bird found in the lowlands of Brazil, Venezuela, and Colombia. This finding provides evidence for a broad host range for P. lutzi that include 2 different orders, Gruiformes and Passeriformes, and also altitudinal expansion of its distribution. The blood stages were compared with the parasites original descriptions, and the sequence of the parasites mitochondrial genome (mtDNA) confirms that P. lutzi is a sister taxa of Plasmodium relictum, as previously proposed.

Leucocytozoon Diversity and Possible Vectors in the Neotropical highlands of Colombia

Studies of the lowland avifauna in the Neotropical Region have shown a paucity of Leucocytozoon species. However, surveys conducted in the Colombian highlands revealed a great diversity of these parasites infecting resident birds. To further investigate the relationship between Leucocytozoon diversity, the potential vectors, and altitudinal distribution, birds from 41 families were sampled at low and high elevations in Colombia. Blood samples were screened by microscopy, and a fragment of cytochrome b was amplified from Leucocytozoon-positive samples. The complete mitochondrial genome was also obtained for each morphospecies of Leucocytozoon. Leucocytozoon species were detected in resident birds, with various degrees of host specificity, at elevations from 2,400 to 3,950 meters above sea level, where five new host-parasite associations were discovered. Phylogenetic analysis based on the cytochrome b fragment suggested that two nominal taxa, L. fringillinarum and L. majoris, are species complexes. Blood sources of Simuliidae revealed generalist-feeding habits that included avian and mammalian hosts. Molecular analysis of parasites in black flies indicated a close relationship with the parasites found in birds. Our investigation provides further evidence that the distribution and transmission of Leucocytozoon species in the Neotropics are influenced by elevation, with the highest prevalence between 2,400 and 3,200 m asl.

Haemoproteus erythrogravidus n. sp. (Haemosporida, Haemoproteidae): Description and molecular characterization of a widespread blood parasite of birds in South America

The great diversity of birds and ecosystems in the Andean mountains has been understudied in terms of their parasite species. We describe a new Haemoproteus parasite, H. (Parahaemoproteus) erythrogravidus infecting Zonotrichia capensis (Rufous-Collared Sparrow) in South America. The description of this blood parasite species is supported by morphological and molecular data based on a fragment of cytochrome b gene (cyt b) and complete mitochondrial genome sequences. The new species is closely related to H. (Parahaemoproteus) coatneyi, and it can be readily distinguished from the latter parasite due to morphology of its blood stages, particularly 1) the formation of a marked protrusion on envelope of infected erythrocytes by the majority of developing gametocytes, a feature which is unique for this Haemoproteus species and 2) the extremely attenuated width of the growing dumbbell-shaped macro- and microgametocytes. Additionally, Haemoproteus erythrogravidus is shown to be a monophyletic taxon that diverges from Haemoproteus coatneyi at the molecular level. We provide the complete mitochondrial DNA genome for both H. coatneyi and H. erythrogravidus. Molecular and morphological evidences indicate that H. erythrogravidus is present in Ecuador and Colombia, and genetic lineages with 100% of identity for the cyt b gene were reported in Chile, Perú, and Venezuela. Our study also indicates that H. erythrogravidus and H. coatneyi are sympatric sister taxa sharing Z. capensis as a host species across its distribution, which could be the result of sympatric speciation or complex biogeographic processes. Further studies on the distribution and evolutionary history of Z. capensis and its parasites H. erythrogravidus and H. coatneyi insight for our better understanding of the factors and dynamics driving parasite speciation.

In vitro development of Haemoproteus columbae (Haemosporida: Haemoproteidae), with perspectives for genomic studies of avian haemosporidian parasites.

The evolutionary origin of wildlife and human malaria parasites (Plasmodium spp.) has been discussed for several decades. The lack of genomic data about species of wildlife haemosporidian parasites related to Plasmodium limits the number of taxa available for phylogenetic analysis. Genomic data about avian parasites of the genus Haemoproteus parasites, the sister genus to Plasmodium are still not available, mainly due to difficulties in obtaining pure DNA of parasites inhabiting nucleated avian host cells. Recent studies show that microgametes of Haemoproteus (Parahaemoproteus) spp. develop in vitro and can be isolated by simple centrifugation, allowing the isolation of pure parasite DNA for genomic studies. However, in vitro development of Haemoproteus (Haemoproteus) spp. has not been investigated, and it is unclear if microgametes of these parasites also can be obtained under in vitro conditions. Here, we provide the first data about the in vitro development of Haemoproteus (Haemoproteus) columbae, a widespread avian haemosporidian parasite, which is specific to pigeons and doves (Columbiformes) and is transmitted by hippoboscid flies (Diptera, Hippoboscidae). In vitro gametogenesis and ookinete development of H. columbae were studied using a strain isolated from a feral Rock Pigeon (Columba livia) in Bogotá-Colombia. The morphological events leading to exflagellation, fertilization and ookinete formation, as well as the rate of development of these stages were followed in vitro at 40 °C, 19 °C and 15 °C for 48 h. Macrogametes, microgametes, zygotes and initial stages of ookinete development were observed in all temperatures, but mature ookinetes were seen only at 40 °C. The largest diversity of sporogonic stages of H. columbae were present at 40 °C however, exflagellation, fertilization of macrogametes and development of immature ookinetes were also observed at 15 °C and 19 °C. Morphological and morphometric features of these stages in vitro were described and illustrated. This study demonstrates a requirement of high temperature for the successful development of mature ookinetes of H. columbae, but not gametes. We show that 1) parasites of the H. (Haemoproteus) subgenus exflagellate in vitro at 15-19 °C, as is the case in H. (Parahaemoproteus) spp. and 2) in vitro exflagellation can be used to obtain pure DNA for genomic studies.