Peer Review #3 of "Blood parasites in Passeriformes in central Germany: prevalence and lineage diversity of Haemosporida (Haemoproteus, Plasmodium and Leucocytozoon) in six common songbirds (v0.1)"

Background: Avian Haemosporida are vector-borne parasites that commonly infect Passeriformes.Molecular analyses revealed a high number of different lineages and lineage specific traits like prevalence and host-specificity, but knowledge of parasite prevalence and lineage diversity in wild birds in Central Germany is still lacking.Results: Blood samples from a total of 238 adult and 122 nestling songbirds belonging to six species were investigated for infections with avian haemosporidian genera and lineages (Haemoproteus spp., Plasmodium spp., Leucocytozoon spp.) and Trypanosoma avium using PCR, targeting the parasite mitochondrial cytochrome b gene and 18S ribosomal RNA.In total, the prevalence in adult birds was 31.3% infected with Haemoproteus, 12.5% with Plasmodium and 71.0% with Leucocytozoon (nestlings excluded).None of the tested birds was infected with Trypanosoma avium.Only in two nestling birds, aged 12-17 days, a Leucocytozoon spp.infection was proven.Among 225 successfully sequenced samples, we found four Haemoproteus, three Plasmodium and 19 Leucocytozoon lineages, including two new Leucocytozoon lineages.Furthermore, we report two new host-lineage associations.Conclusions: As first study investigating avian haemosporidian parasites in Central Germany, we provide new information on genetic diversity of Haemosporida infecting Passeriformes.We show that even with a small sample size new lineages as well as previously unknown linkages between certain lineages and host species can be detected.This may help to elucidate the diversity of lineages as well as lineage-host-connections of avian Haemosporida.