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Fig. 8 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 8. Epimastigotes of G1 and G2 (Clade A) arranged in rosettes in culture. (A) Diff-Quick stained rosettes. (B) Rosettes in fresh wet preparations showing numerous intracellular acidocalcisomes. Scale bars = 10 µm.

Fig. 7 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 7. Light microscopy of Diff-Quick stained blood and culture forms of G1 and G2 (Clade A) (A) Trypomastigote in blood of a woylie naturally infected; (B) slender epimastigote in culture; (C and D) shaped epimastigote in culture; (E) spheromastigote in culture; (F) spheromastigotes dividing in culture. Scale bars = 10 µm.

Fig. 6 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 6. Structures suggestive of amastigotes (arrows) of G2 (Clade A) in heart tissue positive by PCR (H&E stained). Scale bars = (A) 20 µm, (B) 10 µm.

Fig. 5 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 5. Histopathology of two woylies naturally infected with G2 (Clade A) (H&E stained). (A) Multifocal, moderate to severe, chronic, pyogranulomatous myocarditis and (B) endocarditis. (C) Mineralisation of heart tissue. (D) Tongue showing multifocal, moderate, chronic, pyogranulomatous glossitis. (E) Skeletal muscle degeneration. (F) Inflammatory cells around a blood vessel. Scale bars = 20 µm.

Fig. 3 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 3. Phylogenetic relationships of the new trypanosome isolates from Western Australian marsupials based on gGAPDH sequences (~810 bp) using Mr Bayes. The tree was rooted with five sequences as outgroups. Bayesian posterior probabilities are shown at nodes. Bar, 0.07 substitutions per site.

Fig. 10 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 10. Scanning electron micrograph of G2 (Clade A) grown in culture with Vero cells. (A) Trypomastigote invading a cell, with the flagella still external to the cell. (B) Dead cell(s) surrounded by amastigotes and trypomastigotes. Scale bars = 2 µm (A), 4 µm (B).

Fig. 4 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 4. Prevalence of infection with trypanosomes within the different clades in woylies from the stable and declining populations. 95% confidence intervals (95% CI).

Fig. 2 in Trypanosomes genetic diversity, polyparasitism and the population decline of the critically endangered Australian marsupial, the brush tailed bettong or woylie (Bettongia penicillata)

Fig. 2. Phylogenetic analysis of the relationships between Australian trypanosomes based on 18S rDNA sequences. Phylogenetic trees were constructed by the Bayesian method sequences (~1410 bp). (B) Phylogenetic position of shorter 18S rDNA sequences (786 bp) CHA1, TRY1, TRY2, WYA1, WYA2, BDA1, Q3, Q10, GP63 and GP94. Threes were rooted with from Bayesian posterior probabilities are shown at nodes. In red: trypanosome genotypes found in this study. Bar, 0.2 substitutions per site.

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Fig. 1 in Free drug percentage of moxidectin declines with increasing concentrations in the serum of marsupials

Fig. 1. Free drug percentage (%) MOX in serum of koala, kangaroo, and wombat at concentration points 20 pg/μL, 100 pg/μL, and 500 pg/μL. SD; Wombat (20pg/ μL- 0.049; 100pg/μL- 0.062; 500 pg/μL), Koala (20pg/μL- 0.099; 100pg/μL- 0.223; 500pg/μL- 0.038),Kangaroo (100pg/μL- 0.053; 500pg/μL- 0.0049), Possum (100pg/μL- 0.042).

Fig. 4 in Detection of cryptic species of Rugopharynx (Nematoda: Strongylida) from the stomachs of Australian macropodid marsupials

Fig. 4. Phylogenetic relationships of species of Rugopharynx and Rugonema labiatum based on a neighbor-joining analysis of the sequence data of the ITS+ nuclear ribosomal DNA. Values above and below branches represent the NJ and MP bootstrap values (respectively) that were greater than 70%.

Fig. 3 in Detection of cryptic species of Rugopharynx (Nematoda: Strongylida) from the stomachs of Australian macropodid marsupials

Fig. 3. Phylogenetic relationships of species of Rugopharynx and Rugonema labiatum based on a Bayesian analysis of the sequence data of the ITS+ nuclear ribosomal DNA. Values above branches indicate posterior probabilities that were greater than 0.8. Abbreviations of Australian state names are provided in Table 1.

Fig. 2 in Detection of cryptic species of Rugopharynx (Nematoda: Strongylida) from the stomachs of Australian macropodid marsupials

Fig. 2. Morphological buccal capsule types in the genus Rugopharynx. I, simple cylindrical buccal capsule, R. macropodis; IIA, bilobed buccal capsule with subequal divisions, R. epsilon; IIB, bilobed buccal capsule with anterior lobe shorter, R. rufogrisea; III, trilobed buccal capsule, R. longibursaris.

Fig. 1 in Detection of cryptic species of Rugopharynx (Nematoda: Strongylida) from the stomachs of Australian macropodid marsupials

Fig. 1. Localities within Australia at which specimens of Rugopharynx used in this study were collected. Coordinates for each locality are provided in Table 1. 1, Lake Clifton; 2, Waroona; 3, Collie, Wellington Dam; 4, Perup River; 5, Kalgoorlie; 6, Wallerberdina Station; 7, Port Augusta; 8, Ashbourne; 9, Kangaroo Island; 10, Naracoorte; 11, Hattah Lakes National Park; 12, Yan Yean; 13, The Gurdies; 14, Launceston; 15, Emu Flat, Bondo State Forest; 16, Trangie; 17, Grafton; 18, Lamington National Park; 19, Miles; 20, Dawes; 21, Mt Sebastopol; 22, Rockhampton; 23, Winton; 24, Proserpine; 25, Bowen; 26, Magnetic Island; 27, Lake Barrine.

Fig. 5 in Detection of cryptic species of Rugopharynx (Nematoda: Strongylida) from the stomachs of Australian macropodid marsupials

Fig. 5. Molecular phylogeny of species of Rugopharynx and Rugonema labiatum based on a consensus of the BI, NJ and MP trees (Figs. 3 and 4), and the relationships of their hosts. This figure includes species of Macropus (M. agilis, M. antilopinus, M. bernardus) which are not hosts to species of Rugopharynx, as well as M. parma, from which no material could be obtained for genetic studies. Only those species of Petrogale included in this study are shown on the host tree. The morphology of the buccal capsule for each nematode taxon is also shown.

Fig. 3 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 3. Phylogenetic relationships between Trypanosoma sp. ANU2 and other members of the Trypanosoma genus. Maximum likelihood tree is shown. Neighbour-joining and maximumlikelihood bootstrap support followed by Bayesian posterior probability is shown at nodes, respectively. Genbank accession numbers follow species/genotype description. Trypanosoma species/genotypes isolated in Australia are in bold. Scale bar represents substitution per site.

Fig. 6 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 6. Principle Coordinates Analysis (PCoA) plots demonstrating relationship between Trypanosoma spp. ZOTUs and host species. Dissimilarity matrices were generated using sqrt transformed Bray-Curtis distances to show distance between abundance of ZOTUs between host marsupial species; the woylie (blue squares) and brushtail possum (red circles). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 2 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 2. Abundance and diversity map of Trypanosoma copemani genotype 1 (G1) and genotype 2 (G2) positive samples. ZOTUs were sorted into G1 or G2 based on phylogenetic inference shown in rows, while columns are individual marsupial blood samples from infected individuals. The map represents samples separated by host species, which were WOY = woylie, or BTP = brushtail possum. Grayscale indicates number of sequences obtained from that sample as shown in the scale of intensity on the right (log transformed abundance).

Fig. 5 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 5. Trypanosoma spp. polyparasitism in 70 blood samples taken from marsupials in the Upper Warren Region. Marsupial species include: woylie (WOY), brushtail possum (BTP) and chuditch (CHU). Trypanosoma spp. include; C = Trypanosoma copemani, V = T. vegrandis, N = T. noyesi, G = T. gilletti, A = T. sp. ANU2, I = T. irwini, AT = T. sp. AAT, U = unknown, AA = T. avium, and CR = Crithidia spp.

Fig. 4 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 4. Abundance and diversity map of Trypanosoma spp. ZOTUs in different marsupial blood samples assigned to species groups shown in rows, while columns are individual blood samples. Samples are separated by host species including; WOY = woylie (34), CHU = chuditch (3), and BTP = brushtail possum (33). The colour scale indicates increasing number of sequences obtained from that sample, in that species, as shown in the intensity bar on the right. Species include; C = Trypanosoma copemani, V = T. vegrandis, N = T. noyesi, G = T. gilletti, A = T. sp. ANU2, I = T. irwini, AT = T. sp. AAT, U = unknown, AA = T. avium, and CR = Crithidia spp. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig. 1 in Next generation sequencing reveals widespread trypanosome diversity and polyparasitism in marsupials from Western Australia

Fig. 1. Phylogenetic relationships between Trypanosoma spp. ZOTUs assigned to nine species groups compared to 24 representative reference strains downloaded from Genbank. Phytomonas serpens and Leptomonas sp. were used as outgroups. Bayesian analysis was used to produce tree topology and posterior probability is shown at nodes. Scale bar represents substitution per site.