Blood Parasites in Owls with Conservation Implications for the Spotted Owl (Strix occidentalis)

The three subspecies of Spotted Owl (Northern, Strix occidentalis caurina; California, S. o. occidentalis; and Mexican, S. o. lucida) are all threatened by habitat loss and range expansion of the Barred Owl (S. varia). An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp.) or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not been documented in the Spotted Owl. We screened 111 Spotted Owls, 44 Barred Owls, and 387 owls of nine other species for haemosporidian parasites (Leucocytozoon, Plasmodium, and Haemoproteus spp.). California Spotted Owls had the greatest number of simultaneous multi-species infections (44%). Additionally, sequencing results revealed that the Northern and California Spotted Owl subspecies together had the highest number of Leucocytozoon parasite lineages (n = 17) and unique lineages (n = 12). This high level of sequence diversity is significant because only one Leucocytozoon species (L. danilewskyi) has been accepted as valid among all owls, suggesting that L. danilewskyi is a cryptic species. Furthermore, a Plasmodium parasite was documented in a Northern Spotted Owl for the first time. West Coast Barred Owls had a lower prevalence of infection (15%) when compared to sympatric Spotted Owls (S. o. caurina 52%, S. o. occidentalis 79%) and Barred Owls from the historic range (61%). Consequently, Barred Owls on the West Coast may have a competitive advantage over the potentially immune compromised Spotted Owls

Case 3089. Leucocytozoon (Protista, Haemosporida): proposed adoption of Berestneff, 1904 as the author and of Leukocytozoen danilewskyi Ziemann, 1898 as the type species

Volume: 56Start Page: 168End Page: 17

Diversity and phylogeny of mitochondrial cytochrome b lineages from six morphological species of avian Haemoproteus

Species of Haemoproteus (Haemosporida: Haemoproteidae), avian haemosporidians, have traditionally been described based on morphology of their gametocytes and on limited experimental information on their vertebrate host specificity. We investigated to what extent the morphological species are represented by monophyletic groups based on DNA sequence data using 2 different fragment lengths of the cytochrome b (cyt. b) gene. Phylogenetic reconstructions of obtained cyt. b lineages from 6 morphospecies of Haemoproteus showed that all lineages formed monophyletic clusters matching the morphospecies. Comparing our data with a recently published study showed that this is not always the case; the morphospecies H. belopolskyi consists of 2 distinct clusters of lineages that apparently have converged in morphology. However, the overall broad congruence between the molecular and morphological clustering of lineages will facilitate the integration of the knowledge obtained by traditional and molecular parasitology. Mean between morphospecies variation was 10-fold higher than the within species variation (5.5% vs. 0.54%), suggesting that Haemoproteus lineages with a genetic differentiation >5% are expected to be morphologically differentiated in most cases. When investigate the utility of 2 different fragment sizes of the cyt. b gene, the partial, 479-bp, cyt. b protocol picked up all mitochondrial (mt)DNA lineages that are found when using the full cyt. b gene, 1073 bp, suggesting that this protocol is sufficient for identification of most mtDNA lineages. All of the mtDNA lineages were associated with unique alleles when amplification was possible at a nuclear locus, strengthening the hypothesis that the designation of lineages based on mtDNA is largely genome-wide representative. We, therefore, propose the use of a cyt. b fragment of this length as a standard gene fragment for a DNA bar-coding system for avian Haemoproteus species

Plasmodium relictum (lineage SGS1) and Plasmodium ashfordi (lineage GRW2): The effects of the co-infection on experimentally infected passerine birds

The effects of avian malaria parasites of the genus Plasmodium on their hosts are insufficiently understood. This is particularly true for malarial co-infections, which predominant in many bird populations. We investigated effects of primary co-infection of Plasmodium relictum (lineage SGS1) and Plasmodium ashfordi (GRW2) on experimentally infected naive juveniles of siskin Spin us spinus, crossbill Loxia curvirostra and starling Sturnus vulgaris. All siskins and crossbills were susceptible but starlings resistant to both these infections. A general pattern of the co-infections was that heavy parasitemia (over 35% during peaks) of both parasites developed in both susceptible host species. There were no significant effects of the co-infections on mean body mass of the majority of infected birds. Mean haematocrit value decreased approximately 1.5 and 3 times in siskins and crossbills at the peak of parasitemia, respectively. Mortality was recorded among infected crossbills. We conclude that co-infections of P. relictum and P. ashfordi are highly virulent and act synergetically during primary infections in some but not all passerine birds. (C) 2010 Elsevier Inc. All rights reserved

Data from: "Blood transcriptome sequencing of Common Crossbills (Loxia curvirostra) experimentally infected by the avian malaria parasite Plasmodium relictum (lineage SGS1)" in Genomic Resources Notes accepted 1 June 2013-31 July 2013

In order to understand the epidemiology of this avian malaria infections both geographically and across host species there is a need of more polymorphic markers and knowledge about genes expressed during the infection of the host. However, technical difficulties have hindered the development of such molecular markers. The main difficulty is the fact that the host (birds) has nucleated erythrocytes and there is a 52-fold difference in the genome size between the host and the parasite. This has caused problems in trying to isolate and amplify novel nuclear genes of the avian malaria parasites. In order to overcome this difficulty and gain molecular data on nuclear genes we have sequenced blood transcriptomes of common crossbills experimentally infected with Plasmodium relictum SGS1 lineage. During development in the blood, the parasite undergoes several cycles of asexual development (erythrocytic merogony) and is invading numerous host cells and thus is expected to have a high level of gene expression. This data will serve as a resource for researchers interested in developing more molecular markers and investigating genes expressed during the erythrocytic development of avian malaria parasite. With this data we hope to take the first step into the understanding of the mechanism behind the exhibited host generalism as well as understanding how different avian malaria lineages have spread around the globe

Identification and characterization of the merozoite surface protein 1 (msp1) gene in a host-generalist avian malaria parasite, Plasmodium relictum (lineages SGS1 and GRW4) with the use of blood transcriptome

Background: The merozoite surface protein 1 (msp1) is one of the most studied vaccine candidate genes in mammalian Plasmodium spp. to have been used for investigations of epidemiology, population structures, and immunity to infections. However methodological difficulties have impeded the use of nuclear markers such as msp1 in Plasmodium parasites causing avian malaria. Data from an infection transcriptome of the host generalist avian malaria parasite Plasmodium relictum was used to identify and characterize the msp1 gene from two different isolates (mtDNA lineages SGS1 and GRW4). The aim was to investigate whether the msp1 gene in avian malaria species shares the properties of the msp1 gene in Plasmodium falciparum in terms of block variability, conserved anchor points and repeat motifs, and further to investigate the degree to which the gene might be informative in avian malaria parasites for population and epidemiological studies. Methods: Reads from 454 sequencing of birds infected with avian malaria was used to develop Sanger sequencing protocols for the msp1 gene of P. relictum. Genetic variability between variable and conserved blocks of the gene was compared within and between avian malaria parasite species, including P. falciparum. Genetic variability of the msp1 gene in P. relictum was compared with six other nuclear genes and the mtDNA gene cytochrome b. Results: The msp1 gene of P. relictum shares the same general pattern of variable and conserved blocks as found in P. falciparum, although the variable blocks exhibited less variability than P. falciparum. The variation across the gene blocks in P. falciparum spanned from being as conserved as within species variation in P. relictum to being as variable as between the two avian malaria species (P. relictum and Plasmodium gallinaceum) in the variable blocks. In P. relictum the highly conserved p19 region of the peptide was identified, which included two epidermal growth factor-like domains and a fully conserved GPI anchor point. Conclusion: This study provides protocols for evaluation of the msp1 gene in the avian malaria generalist parasite P. relictum. The msp1 gene in avian Plasmodium shares the genetic properties seen in P. falciparum, indicating evolutionary conserved functions for the gene. The data on the variable blocks of the gene show that the msp1 gene in P. relictum might serve as a good candidate gene for future population and epidemiological studies of the parasite

Molecular characterization of haemosporidian parasites (Haemosporida) in yellow wagtail (Motacilla flava), with description of in vitro ookinetes of Haemoproteus motacillae

In studies of haemosporidian systematics and taxonomy, a combination of microscopic examination and molecular identification has been recommended. The yellow wagtail (Motacilla flava) is a frequently used species in studies of host-parasite interactions; blood parasites of this bird have been well studied using microscopic examination of blood films, but data on molecular characterization of the parasites are scattered. Here we present the first study linking several haemosporidian cyt b sequences with their morphospecies parasitizing the yellow wagtail, combining light microscopy and nested PCR methods. The lineage hYWT1 of Haemoproteus motacillae was the most prevalent parasite; it was recorded in 38% of the birds. The latter lineage and the lineages hYWT2, hYWT3, hYWT5 and hYWT6 form a well-supported clade on the phylogenetic tree and likely represent intraspecific genetic variation of H. motacillae, with genetic divergence of 0.3-1.5 % among these lineages. Microscopic examination of smears prepared during an in vitro experiment, which was designed for observation of developing ookinetes, showed that H. motacillae ookinetes were present 5 hours after exposure of blood to air at 18-20 degrees C. Ookinetes of this parasite belong to a group of large Haemoproteus spp. ookinetes (19.9 mu m in length on average). Illustrations of the parasites are given. Two of reported Plasmodium lineages, pCOLL1 and pYWT4 are phylogenetically closely related to the widespread host generalist parasites Plasmodium relictum (cyt b lineages pSGS1, pGRW4 and pGRW11), but it was not possible to establish this morphologically. This study contributes to the molecular identification of avian haemosporidian parasites and provides information on morphology of H. motacillae ookinetes, which is additional information assisting the microscopic identification of this species

Comparison of mitochondrial cytochrome b lineages and morphospecies of two avian malaria parasites of the subgenera Haemamoeba and Giovannolaia (Haemosporida : Plasmodiidae)

PCR-based methods have been increasingly used in diagnosis of parasitic diseases. Over 40 morphospecies of avian malaria parasites of the genus Plasmodium have been described. However, only nine of them have been identified on the level of their mitochondrial cytochrome b (cyt b) gene lineages, which are frequently used in molecular biology studies of avian blood haemosporidian parasites. In this study, we linked two common mitochondrial cyt b lineages P-SGS1 and P-TURDUS1 with their morphospecies. Light infections with two species of malaria parasites of the subgenera Haemamoeba and Giovannolaia were isolated from naturally infected adult Hawfinches, Coccothraustes coccothraustes Linnaeus, on the Curonian Spit in the Baltic Sea. These parasites were inoculated to naive juveniles of the Common Crossbill, Loxia curvirostra Linnaeus, and House Sparrow, Passer domesticus Linnaeus. Heavy parasitemia of Plasmodium (Haemamoeba) relictum Grassi & Feletti, 1891 ( lineage P-SGS1) and Plasmodium (Giovannolaia) circumflexum Kikuth, 1931 (P-TURDUS1) developed in the subinoculated Common Crossbills and House Sparrows, respectively, which enabled the detailed illustration of all main blood stages of these parasites and the deposition of their voucher specimens. The parasites of both lineages are actively transmitted in Europe and inhabit a broad range of avian hosts. Lineages closely related to P. relictum and P. circumflexum were identified. This study contributes to establishing of combined PCR-based and morphological identification of avian malaria parasites