Table 1.
Blood samples collected from Australian marsupials screened for piroplasms in this study.
Table 2.
Samples positive for piroplasm 18S rDNA from wildlife and ticks utilised in this study.
Fig 1.
Phylogenetic analysis of newly reported Theileria spp. and other piroplasms based on the 18S rRNA locus.
(A) Phylogenetic relationships among piroplasm 18S rDNA sequences (~ 1.3 kb) were inferred using the maximum likelihood method. (B) Maximum likelihood analysis of piroplasms based on a 800bp alignment to illustrate the position of newly reported organisms in the context of all known Theileria spp. recorded in Australia to date. Names in bold represent sequences retrieved from GenBank that were not included in the main tree (Fig 1, A) due to their shorter length. GenBank accession codes are shown in parentheses. Bootstrap values (>60%) based on 500 replicates are indicated at the left of each node. * indicates shorter sequences (~ 800 bp) obtained in this study using a nested PCR as described by Jefferies et. al. (2007). All other new sequences (~1.3 kb) were obtained using primers Nbab_1F and 18S ApiR (Greay et al., 2018).
Fig 2.
Phylogenetic analysis of cox3 nucleotide sequences from Australian piroplasms and other piroplasms for which cox3 sequences are available in GenBank.
Phylogenetic relationships among sequences (~ 600 bp) were inferred using the maximum likelihood method. Names in bold represent sequences obtained in the present study from marsupial blood samples (Table 1) and positive piroplasm 18S rDNA samples obtained during previous studies (Table 2). Symbols at the left of each sequence correspond to the clades identified in the phylogenetic analysis at the 18S rRNA locus (Fig 1).
Fig 3.
Phylogenetic analysis of cytB nucleotide sequences from Australian piroplasms and other piroplasms for which cytB sequences are available in GenBank.
Phylogenetic relationships among sequences (~1.1 kb) were inferred using the maximum likelihood method. Names in bold represent sequences obtained in the present study from marsupial blood samples (Table 1) and positive piroplasm 18S rDNA samples obtained during previous studies (Table 2). Symbols at the left of each sequence correspond to the clades identified in the phylogenetic analysis at the 18S rRNA locus (Fig 1).
Fig 4.
Approximate location (where known) of Theileria spp. from marsupials and ticks recorded in Australia to date.
Icons in red represent the spatial distribution and host-range of novel Theileria spp. identified in the present study.
Fig 5.
Microscopic detection of intra-erythrocytic ring-shaped piroplasms in blood films from.
(A) eastern quoll (Dasyurus viverrinus) (TB105); (B) eastern bettong (Bettongia gaimardi) (TB27); (C) quokka (Setonix brachyurus) (Q1051); (D) quokka (Q2031a); (E) Gilbert’s potoroo (Potorous gilbertii) (P92); (F) quokka (Q1416). Scale bars represent 10 μm.