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

Abstract

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.

Polymerase chain reaction-based identification of Plasmodium (Huffia) elongatum, with remarks on species identity of haemosporidian lineages deposited in GenBank

“…However, only 11 morphospecies of avian Plasmodium have been linked to these lineages (Palinauskas et al 2007;Valkiūnas et al 2007b). Two identical sequences (DQ368381 and DQ659588) can be used for the molecular diagnosis of P. elongatum infection.…”

“…If the information about the relationships of lineages and morphospecies currently present in GenBank is correct, the use of morphological and life history characters in the taxonomy of haemosporidian parasites should be discouraged. However, some recent studies have shown good concordance between traditional taxonomy of haemosporidians and molecular clustering of their lineages (Hellgren et al 2007;Martinsen et al 2007;Perkins et al 2007;Palinauskas et al 2007;Valkiūnas et al 2007a, b).…”

“…Molecular diagnostic techniques are sensitive in detecting blood parasites, especially during low parasitaemias Ricklefs et al 2004;Sehgal et al 2006;Valkiūnas et al 2006;Hellgren et al 2007). However, only DNA sequences, which are clearly linked with their morphospecies, can be used for this purpose (Palinauskas et al 2007). Unfortunately, sequences of haemosporidian parasites, which have been misidentified even at the level of genera, are present in GenBank (Valkiūnas et al 2007b).…”

“…Because identification of P. elongatum is of practical and theoretical significance, the main aim of this study was to develop a method for identification of this malaria parasite based on partial sequences of its mitochondrial cyt b gene and using the polymerase chain reaction (PCR), which has been successfully used in evolutionary biology studies of haemosporidian blood parasites (Escalante et al 1998;Fallon et al 2003;Bensch et al 2004;Pérez-Tris and Bensch 2005;Križanauskienė et al 2006;Hellgren et al 2007;Palinauskas et al 2007;Perkins et al 2007). To achieve this aim, we used experimental infections with high parasitaemia in naive birds, as recommended by Palinauskas et al (2007) and Valkiūnas et al (2007b).…”

“…To achieve this aim, we used experimental infections with high parasitaemia in naive birds, as recommended by Palinauskas et al (2007) and Valkiūnas et al (2007b). This permitted the clear morphological identification of P. elongatum, the deposition of voucher specimens of its blood stages and the determination of mitochondrial cyt b lineages, which can be used for molecular diagnosis of P. elongatum infection.…”