Morphological and molecular characterization of Eimeria haematopusi n. sp. (Apicomplexa: Eimeriidae) in an Australian Pied Oystercatcher (Haematopus longirostris) (Aves: Charadriiformes).

A novel Eimeria Schneider, 1875 species is described from an Australian pied oystercatcher Haematopus longirostris Vieillot, in Western Australia. The pied oystercatcher was admitted to the Kanyana Wildlife Rehabilitation Centre (KWRC), Perth, Western Australia in a poor body condition, abrasion to its right hock and signs of partial delamination to its lower beak. Investigation into potential medical causes resulted in a faecal sample being collected and screened for gastrointestinal parasites. Unsporulated coccidian oocysts were initially observed in the faeces and identified as Eimeria upon sporulation. The sporulated oocysts (n = 20) are ellipsoidal, 20-21 × 12-13 µm in shape and have thick bi-layered walls which are c.2/3 of the total thickness. Micropyle is present, robust and protruding, and occasionally has a rounded polar body attached to the micropyle. Within the oocyst, a residuum, in addition, two to five polar granules are present. There are four ellipsoidal sporocysts 9-11 × 5-6 µm with flattened to half-moon shaped Stieda bodies. Sub-Stieda body and para-Stieda body are absent. The sporocysts contain sporocyst residuums composed of a few spherules scattered among the sporozoites. Within the sporozoites, anterior and posterior refractile bodies are present, but the nucleus is indiscernible. To further characterise the novel Eimeria species from H. longirostris, molecular analysis was conducted at the 18S ribosomal RNA locus, using PCR amplification and cloning. Two cloned sequences from the novel Eimeria were compared with those from other Eimeria spp. with the highest genetic similarity of 97.6% and 97.2% from Clone 1 and 2, respectively with Eimeria reichenowi (AB544308) from a hooded crane (Grus monacha Temminck) in Japan. Both sequences grouped in a clade with the Eimeria spp. isolated from wetland birds, which include Eimeria paludosa (KJ767187) from a dusky moorhen (Gallinula tenebrosa Gould) in Western Australia, Eimeria reichenowi (AB544308) and Eimeria gruis (AB544336) both from hooded cranes. Based on the morphological and molecular data, this Eimeria sp. is a new species of coccidian parasite and is named Eimeria haematopusi n. sp. after its host H. longirostris.

Morphological and genetic characterization of the first Isospora species (I. lugensae n. sp.) from a Kerguelen petrel (Lugensa brevirostris).

A new coccidian species, Isospora lugensae n. sp., was described from a single Kerguelen petrel (Lugensa brevirostris). Sporulated oocysts (n = 25) were characterized as subspheroidal to ellipsoidal measuring 24-25 µm × 21-23 µm (24.8 × 22.2 µm) in length/width (L/W), respectively, with a ratio of 1.07-1.14 µm (1.12). They contained a bi-layered wall with a thickness of 0.8-1.2 µm (1.0) and the outer layer smooth, with c.2/3 of total thickness. The oocyst contained two polar granules with both micropyle and oocyst residuum absent. Ovoidal sporocysts (n = 25) measured 15-16 µm × 10-11 µm (15.7 × 10.8 µm) in L/W, with a ratio of 1.41-1.49 µm (1.46). A flattened to knob-like Stieda body was present (c.0.5 µm deep × 2.5 µm wide) as well as a rounded to trapezoidal sub-Stieda (c.1.5 µm deep × 3.0 µm wide); however, no para-Stieda body was detected. The sporocyst residuum was composed of scattered spherules of different sizes, while vermiform sporozoites contained a refractile body, nucleus and visible striations. Analysis of the full-length mitochrondrial (mtDNA) genome revealed 3 protein-coding genes, (CytB, COI and COIII), 18 LSU and 14 small subunit (SSU) rDNA fragments, without transfer RNA genes with a total length of 6257 bp. Phylogenetic analysis of genomic SSU ribosomal sequences indicated that Isospora lugensae n. sp. is genetically similar to Eimeria reichenowi, isolated from a red-crowned crane (Grus japonensis) from Japan, with a 96.6% homology. The mtDNA sequence is most similar to Isospora serinuse with a 95.8% genetic similarity. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in a Kerguelen petrel.

Morphological and genetic characterization of Eimeria chalcoptereae n. sp. (Apicomplexa: Eimeriidae) in a common bronzewing pigeon (Phaps chalcoptera) (Latham, 1790) in Western Australia.

A new Eimeria species is described from a common bronzewing pigeon (Phaps chalcoptera) (Latham, 1790) in Western Australia. Sporulated oocysts of Eimeria chalcoptereae n. sp. (n = 30) are subspheroidal, 22-25 × 21-24 (23.5 × 22.6) µm; length/width (L/W) ratio 1.0-1.1 (1.04) µm. Wall bi-layered, 1.0-1.4 (1.2) µm thick, outer layer smooth, c.2/3 of total thickness. Micropyle barely discernible. Oocyst residuum is absent, but 2 to 3 small polar granules are present. Sporocysts (n = 30) ellipsoidal, 13-14 × 7-8 (13.5 × 7.2) µm; L/W ratio 1.8-2.0 (1.88). Stieda body present, flattened to half-moon-shaped, 0.5 × 2.0 µm; sub-Stieda present, rounded to trapezoidal, 1.5 × 2.5 µm; para-Stieda body absent; sporocyst residuum present, usually as an irregular body consisting of numerous small granules that appear to be membrane-bound. Sporozoites vermiform, with a robust refractile body and centrally located nucleus. Isolated Eimeria oocysts were analysed at the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. Analyses revealed that Eimeria chalcoptereae n. sp. shared the highest number of molecular features with an Eimeria sp. previously identified from a domestic pigeon in Australia (KT305927-29), with similarities at these three loci of 98.53%, 97.32% and 94.93%, respectively. According to morphological and molecular analysis, the isolated coccidian parasite is a new species of Eimeria named Eimeria chalcoptereae n. sp. after its host, the common bronzewing pigeon (Phaps chalcoptera) (Columbiformes: Columbidae) (Latham, 1790).

Molecular and morphological analysis of a Caryospora-like isolate (Apicomplexa: Eimeriidae) from the magpie-lark (Grallina cyanoleuca) (Latham, 1801) in Western Australia.

A new Caryospora-like isolate is described from a magpie-lark (Grallina cyanoleuca) in Western Australia. Sporulated oocysts of the Caryospora-like isolate (n = 35) are subspherical with a shape index of 1.13 ((21.5 (19.7-23.6) × 19.0 (18.1-19.8) µm). The bilayered oocyst wall is smooth. Micropyle, polar granule and oocyst residuum are absent. The sporocyst is ellipsoidal, 18.9 (17.2-20.8) × 12.3 (11.9-12.8) µm, with a shape index (length/width) of 1.54. The sporocyst wall is bilayered. Stieda and substieda bodies are present, the Stieda body is small and flattened and the substieda is trapezoidal. Sporocyst with eight sporozoites arranged head to tail. The sporozoites are vermiform, 18.9 (17.2-20.8) × 12.3 (11.9-12.8) µm and have striations at the anterior end. Each sporozoite has both anterior and posterior refractile bodies. A sporocyst residuum is present. Molecular characterization of the isolated Caryospora-like oocysts was conducted at the 18S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. At the 18S rRNA locus, the Caryospora-like isolate exhibited 88.8% to 96.5% similarity with other Caryospora spp. from different hosts. At the COI locus, it showed 91.5% similarity to Caryospora cf. bigenetica JB-2013 (KF859856) from the rattlesnake, Sistrurus catenatus.

Isospora coronoideae n. sp. (Apicomplexa: Eimeriidae) from the Australian raven (Corvus coronoides) (Passeriformes: Corvidae) (Linnaeus, 1758) in Western Australia.

A new Isospora (Apicomplexa: Eimeriidae) species is described from an Australian raven (Corvus coronoides) in Western Australia. Sporulated oocysts (n = 21) are ovoid, 21.2 (18.4-23.9) µm in length and 18.8 (16.9-20.6) µm in width, with a shape index of 1.13. The bi-layered oocyst wall is smooth and colourless, 1.2 µm thick. A polar granule and oocyst residuum is present, but the micropyle is absent. The sporocysts are ovoid-shaped, 16.3 (13.7-18.9) × 10.7 (8.4-12.9) µm, with a shape index (length/width) of 1.52. Stieda and substieda bodies are present, the Stieda body being small and hemidome-shaped and the substieda being indistinct. Each sporocyst with four vermiform sporozoites arranged head to tail. The sporozoites are crescent-shaped, 9.0 (8.9-9.2) × 2.7 (2.3-3.0) µm, with a shape index (length/width) of 3.33. The sporocyst residuum is present. The isolated oocysts had different morphological characteristics when compared with all known Isospora spp. The coccidian parasite was analysed at the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) loci. At the 18S locus, I. coronoideae n. sp. exhibited 98.9% similarity to I. neochmiae from a captive-bred red-browed finch (KT224380) and Isospora sp. from domestic pigeons (Columba livia domestica) (AB757860), 98.5% similarity to I. gryphoni (AF080613) from an American goldfinch and 98.3% similarity to I. manorinae (KT224379) from a yellow-throated miner. At the 28S locus, it exhibited 95.4% and 94.8% similarity to I. manorinae (KT224381) and I. anthochaerae (KF766053), respectively. At the COI locus, it exhibited 99.8% and 99.7% similarity to I. butcherae (KY801687) and I. neochmiae (KT224378), respectively. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora coronoideae n. sp. after its host, the Australian raven (Corvus coronoides) (Passeriformes: Corvidae) (Linnaeus, 1758).

Further characterisation of Leucocytozoon podargii in wild tawny frogmouths (Podargus strigoides) in Western Australia.

The present study assessed the prevalence and morphology of Leucocytozoon podargii from wild tawny frogmouths (Podargus strigoides) in Western Australia (WA) and genetically characterised the cytochrome b gene (cyt b) of L. podargii in wild tawny frogmouths from WA and Queensland (QLD). The prevalence of L. podargii in wild tawny frogmouths from WA was 93.3% (14/15; 95% CI, 68.1-99.8%). The morphological characters of L. podargii from WA were similar to L. podargii from QLD: the gametocytes were round-oval shape, approximately 8-12 µm in diameter; the macrogametocytes were 12.4 µm in diameter; microgametocytes were 10.4 µm in diameter; and the ratio of macrogametocytes and microgametocytes was 3:2. Sequence analysis of partial cyt b gene fragments revealed that L. podargii sequences isolated from wild tawny frogmouths in WA shared the highest similarity (99.8% at nucleotide level and 100% at protein level) with L. podargii isolated from wild tawny frogmouths in QLD. The mitochondrial 18S rRNA gene of L. podargii gametocytes was quantified using droplet digital PCR (ddPCR), and the highest gametocyte load was detected in the lung. This finding corresponds to the results of the histological study. Based on the morphological and molecular studies, it was concluded that the Leucocytozoon parasite identified from wild tawny frogmouths in WA is consistent with L. podargii from wild tawny frogmouths in QLD, and the present study has genetically characterised two different L. podargii genotypes (QLD and WA) for the first time.

Morphological and molecular characterisation of Isospora butcherae n. sp. in a silvereye (Zosterops lateralis) (Latham, 1801).

A new Isospora (Apicomplexa:Eimeriidae) species is described from a silvereye (Zosterops lateralis) in Western Australia. Sporulated oocysts of this species are spherical, 24.2 (23.1-25.2) × 23.3 (22.8-23.9) µm, with a shape index (length/width) of 1.02, and with a smooth bi-layered oocyst wall, 1.2 µm thick (outer layer 0.9 µm, inner 0.3 µm). A polar granule is present, but the oocyst residuum and a micropyle are absent. The ovoid-shaped sporocysts are 16.1 (15.7-17.3) × 10.5 (15.7-17.3) µm and have a shape index of 1.53. A hemidome-shaped Stieda and a rectangular-shaped substieda body are present. A sporocyst residuum is present and composed of numerous granules of different sizes scattered among the sporozoites. The oocysts from this isolate are morphologically different from those of all known Isospora spp. This coccidian parasite was molecularly characterised at the 18S, 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, based on 1210 bp of sequence, this new isolate exhibited 99.9, 99.8, 99.7 and 99.5% similarity to I. sp. MAH-2013a (KF648870) from a superb starling (Lamprotornis superbus) in Canada, I. sp. MS-2003 (AY33157) from a Southern cape sparrow (Plocepasser mahali) in America, I. sp. Tokyo (AB75786) from Japan and I. sp. respectively. Further analysis of a subgroup of 300 bp long 18S sequences (n = 11), including I. anthochaerae and the other three Isospora characterised from birds in Western Australia, revealed that I. butcherae n. sp. exhibited 98.3% similarity to both I. sp. MAH-2013a (KF648870) and I. MS-2003 (AY33171). At the 28S locus, this new isolate exhibited 97.3% similarity with I. sp. MS-2003 from a California towhee (Melozone crissalis). At the COI locus, this new isolate exhibited 99.8% similarity to I. neochmiae from a red-browed finch. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora butcherae n. sp. after Mrs. June Butcher for her lifelong dedication as a wildlife rehabilitator. Graphical abstract ᅟ.

Morphological and molecular characterization of Choleoeimeria pogonae n. sp. coccidian parasite (Apicomplexa: Eimeriidae, 1989, Paperna and Landsberg) in a western bearded dragon (Pogona minor minor).

A new species, Choleoeimeria pogonae n. sp. is described from a Western bearded dragon (Pogona minor minor) in Western Australia. Sporulated oocysts (n = 48) were cylindroidal in shape. Oocyst length, 27.0 (26.0-28.3) µm, oocyst width, 15.2 (14.0-16.5) µm, oocyst length/width ratio (L/W) 1.8 (1.6-1.9), each with 4 sporocysts (Eimeria-like) and a polar granule, but lacking a micropyle and oocyst residuum. Sporocysts are ovoidal in shape, sporocyst length, 10.0 (9.0-11.0) µm, sporocyst width 8.5 (7.0-9.5) µm, sporocyst L/W ratio, 1.2 (1.1-1.3). Stieda, substieda and parasubstieda bodies were all absent. Molecular analysis was conducted at the 18S rRNA and cytochrome c oxidase I (COI) loci. Phylogenetic analysis of 18S sequences revealed that C. pogonae n. sp. grouped together with another four Choleoeimeria spp. and exhibited 99.1%-99.4% genetic similarity. At the COI locus, C. pogonae n. sp. was in an independent clade and had the highest similarity (80.4%) to Eimeria cf. mivati from a chicken (Gallus gallus domesticus). According to the morphological and molecular data, this isolate is a new species of coccidian parasite. This study further supports the taxonomy of Choleoeimeria spp. as a new genus based on molecular phylogenetic analysis.

Morphological and molecular characterization of Isospora neochmiae n. sp. in a captive-bred red-browed finch (Neochmia temporalis) (Latham, 1802).

A new Isospora (Apicomplexa: Eimeriidae) species is described from a single red-browed finch (Neochmia temporalis) (subspecies N. temporalis temporalis), that was part of a captive population in Western Australia. Sporulated oocysts of this isolate are spherical, 18.3 (18.2-18.9) × 18.2 (18.2-18.6) µm, with a shape index (length/width) of 1.0; and a smooth and bilayered oocyst wall, 1.2 µm thick (outer layer 0.9 µm, inner 0.3 µm). A polar granule is present, but the oocyst residuum and a micropyle are absent. The sporocysts are ovoid-shaped, 13.3 (9.5-16.4) × 8.6 (6.8-10.0) µm, with a shape index of 1.5. An indistinct Stieda body is present, but the substieda body is absent. A sporocyst residuum is present and composed of numerous granules of different size scattered among the sporozoites. Morphologically, the oocysts from this isolate are different from those of all known valid Isospora spp. Molecular analysis was conducted at 4 loci; the 18S and 28S ribosomal RNA (rRNA), the mitochondrial cytochrome oxidase (COI) gene and the heat shock protein 70 (hsp70) gene. At the 18S locus, this new isolate exhibited 99.9%, 99.8%, 99.7%, and 99.5% similarity to I. sp. MAH-2013a from a superb starling (Lamprotornis superbus), I. MS-2003 from a Southern cape sparrow (Passer melanurus), I. sp. Tokyo from a domestic pigeon (Columba livia domestica) and I. MS-2003 from a Surinam crested oropendula (Psarocolius decumanus). At the 28S locus, this new isolate exhibited 99.7% similarity to both an Isospora sp (MS-2003) from a Northern house sparrow (Passer domesticus) and an Isospora sp. (MS-2003) from a Southern cape sparrow. At the COI locus, this new isolate exhibited 98.9% similarity to an Isospora sp. ex Apodemus flavicollis. At the hsp70 locus, this new isolate exhibited 99% similarity to isolate MS-2003 (AY283879) from a wattled starling (Creatophora cinerea). Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora neochmiae n. sp. after its host, the red-browed finch (Neochmia temporalis).

Morphological and molecular characterization of Eimeria labbeana-like (Apicomplexa:Eimeriidae) in a domestic pigeon (Columba livia domestica, Gmelin, 1789) in Australia.

An Eimeria species is described from a domestic pigeon (Columba livia domestica). Sporulated oocysts (n = 35) were subspherical, with a smooth bi-layered oocyst wall (1.0 µm thick). Oocysts measured 20.2 × 16.1 (22.0-18.9 × 15.7-18.9) µm, oocyst length/width (L/W) ratio, 1.38. Oocyst residuum and a polar granule were present. The micropyle was absent. Sporocysts are elongate-ovoid, 13.0 × 6.1 (14.5-12.5 × 5.5-7.0) µm, sporocyst L/W ratio, 2.13 (2.0-2.2), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 banana-shaped sporozoites, 12.3 × 3.5 (11.8-13.0 × 3.3-3.6) µm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus was located immediately anterior to the posterior refractile body. Molecular analysis was conducted at three loci; the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18S locus, the new isolate shared 98.0% genetic similarity with three Isospora isolates from Japan from the domestic pigeon (Columba livia domestica). At the 28S locus, it grouped separately and shared 92.4% and 92.5% genetic similarity with Isospora anthochaerae (KF766053) from a red wattlebird (Anthochaera carunculata) from Australia and an Isospora sp. (MS-2003 - AY283845) from a Himalayan grey-headed bullfinch (Pyrrhula erythaca) respectively. At COI locus, this new isolate was in a separate clade and shared 95.6% and 90.0% similarity respectively with Eimeria tiliquae n. sp. from a shingleback skink in Australia and an Eimeria sp. from a common pheasant (Phasianus colchicus) from America. Based on the morphological data, this isolate is most similar to Eimeria labbeana. As no molecular data for E. labbeana is available and previous morphological data is incomplete, we refer to the current isolate as E. labbeana-like.

Morphological and molecular characterization of Isospora manorinae n. sp. in a yellow-throated miner (Manorina flavigula wayensis) (Gould, 1840).

A new Isospora (Apicomplexa:Eimeriidae) species is described from a single yellow-throated miner bird (Manorina flavigula) (subspecies M. f. wayensis) in Western Australia. Sporulated oocysts (n = 32) of this isolate are spherical to subspherical, 22.8 (20.3-23.8) × 18.3 (17.7-18.7) µm, with a shape index (length/width) of 1.25 (1.2-1.3); and a smooth and bilayered oocyst wall, 1.3 µm thick (outer layer 0.9 µm, inner 0.4 µm). A polar granule is present, but the micropyle and oocyst residuum are absent. The sporocysts are lemon-shaped, 15.5 (14.6-15.8) × 9.5 (9.5-10.2) µm, with a shape index of 1.6. Stieda and substieda bodies are present, the Stieda body being knob-like and the substieda body being subspherical-shaped. A sporocyst residuum is present and composed of numerous granules of different size scattered among the sporozoites, a spheroid or subspheroid refractile body is present in the sporozoite. Morphologically, the oocysts from this isolate are different from those of all known valid Isospora spp. Molecular analysis was conducted at 3 loci; the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, this new isolate exhibited 99.2% similarity to Isospora gryphoni and three other Isospora spp. Further analysis of a subgroup of 300 bp long 18S sequences (8), including Isospora anthochaerae was conducted. This new isolate grouped in a clade with I. anthochaerae and exhibited 99.3% similarity. At the 28S locus, this new isolate grouped with I. anthochaerae with which it shared 99.1% similarity. At the COI locus, this new isolate exhibited 96.8% similarity to Isospora sp. JCI-2015 from a spectacled warbler (Sylvia conspicillata) in Spain. Further analysis from a subgroup of shorter COI sequences (n = 13) was performed and this new isolate exhibited 99.1% similarity to I. anthochaerae. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora manorinae n. sp. after its host, the yellow-throated miner (Manorina flavigula wayensis).

Morphological and molecular characterization of Eimeria haematodi, coccidian parasite (Apicomplexa: Eimeriidae) in a rainbow lorikeet (Trichoglossus haematodus).

Eimeria haematodi was first described in 1977 from the rainbow lorikeet (Trichoglossus haematodus) in Papua New Guinea. In the present study, we re-describe this coccidian species morphologically and molecularly from a rainbow lorikeet bird in Western Australia (WA). The oocysts were ovoid to slightly piriform and measured 28.5-37.8 by 25.8-33.0 µm (33.3 by 28.1 µm). Oocyst wall was approximately 1.5 µm thick and bilayered. Micropyle (5-7 µm) and oocyst residuum (8.0-10.0 µm) present; polar granule was absent. Sporocysts ellipsoidal, 11.8-13.6 by 8.0-9.6 µm (12.2 by 8.3 µm), with thin convex Stieda body and granular sporocyst residuum (4.0-5.0 µm). Molecular characterization of E. haematodi was conducted at 18S ribosomal RNA and the mitochondrial cytochrome oxidase gene (COI) loci. At the 18S ribosomal RNA locus, E. haematodi shared 98.1% genetic similarity to E. alabamensis from cattle in New South Wales, Australia. At COI locus, E. haematodi was closest (92.3% similarity) to E. praecox from domestic chickens (Gallus gallus domesticus) from Canada and China.

Eimeria collieie n. sp. (Apicomplexa:Eimeriidae) from the western long-necked turtle (Chelodina colliei).

A new species, Eimeria collieie n. sp., is described from the western long-necked turtle (Chelodina colliei). Sporulated oocysts (n = 35) are spherical to subspherical, with colourless single layer oocyst wall, 0.6 ± 0.2 (0.4-0.7) µm thick. Oocyst with elongated ellipsoid sporocysts. Oocyst length, 29.8 ± 0.4 (28.2-31.0) µm; oocyst width, 29.4 ± 0.3 (28.0-30.8) µm; oocyst length/width (L/W) ratio, 1.0 ± 0.03 (1.0-1.05). Micropyle, oocyst residuum and polar granule were absent. Sporocysts with sporocyst residuum and 2 sporozoites. Sporocyst length, 21.6 ± 0.4 (21.2-22.0) µm; sporocyst width, 6.0 ± 0.3 (5.7-6.3) µm; sporocyst L/W ratio, 3.6 ± 0.2 (3.4-3.8). Stieda, parastieda and substieda bodies were absent. Sporozoite length, 14.0 ± 0.2 (13.8-14.2) µm; sporozoite width, 2.6 ± 0.2 (2.4-2.8) µm; sporozoite L/W ratio, 5.46 ± 0.10 (5.4-5.6). Molecular analysis was conducted at three loci: the 18S and 28S ribosomal RNA (rRNA), and the mitochondrial cytochrome oxidase gene (COI). At the 18S rRNA locus, E. collieie n. sp. shared 96.4% and 98.3% genetic similarity to E. ranae (GenBank accession number: EU717219) and E. arnyi (AY613853) respectively. At the 28S rRNA locus, E. collieie n. sp. shared 91.6% genetic similarity to E. papillata (GenBank accession number: GU593706) and phylogenetic analysis at this locus placed E. collieie n. sp. in aseparateclade. At the COI locus, E. collieie n. sp. shared 92.7% genetic similarity to Eimeria setonicis (GenBankaccession number: KF225638) from a quokka (Setonix brachyurus) in Western Australia. Reptile-derived sequences were not available for the 28S rRNA and the COI loci. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that, to date, has only been found in western long-necked turtles.

Isospora serinuse n. sp. (Apicomplexa: Eimeriidae) from a domestic canary (Serinus canaria forma domestica) (Passeriformes: Fringillidae) in Western Australia.

A new species, Isospora serinuse n. sp., (Apicomplexa:Eimeriidae) is described from a single domestic canary (Serinus canaria forma domestica) (subspecies S. c. domestica) in Western Australia. Sporulated oocysts of Isospora serinuse n. sp. are spherical or subspherical, 25.5 (24.4-27.0) × 23.5 (22.0-24.8) µm, with a shape index (length/width) of 1.09; and a smooth bilayered oocyst wall, 1.2 µm thick (outer layer 0.9 µm, inner 0.3 µm). A polar granule is present, but a micropyle and oocyst residuum are absent. The sporocysts are lemon-shaped, 18.9 (17.8-20.2) × 11.8 (10.6-13.0) µm, with a shape index of 1.6. Stieda and substieda bodies are present, the Stieda body being a small crescent shape and the substieda being indistinct. Each sporocyst with four vermiform sporozoites arranged head to tail. A sporocyst residuum is present and composed of numerous granules of different sizes that are scattered among the sporozoites. Morphologically, the oocysts of Isospora serinuse n. sp. were different from those of all known valid Isospora spp. Molecular analysis was conducted at 3 loci: the 18S and 28S ribosomal RNA and two separate regions of subunit I of the mitochondrial cytochrome oxidase (COI) gene (designated COIa and COIb). At the 18S locus, Isospora serinuse n. sp. exhibited 97.5% similarity to Isospora sp. Tokyo from a domestic pigeon (Columba livia domestica) in Japan. At the 28S locus, I. serinuse n. sp. exhibited 94.9% similarity to Isospora anthochaerae n. sp. from a red wattlebird (Anthochaera carunculata) in Australia. At the COIa locus, I. serinuse n. sp. exhibited 95.7% similarity to Isospora sospora sp. ex Apodemus flavicollis from a yellow-necked mouse and Isospora gryphoni from an American goldfinch (Carduelis tristis) respectively. At the COIb locus, I. serinuse n. sp. exhibited 96.7% similarity to an Isospora (iSAT4) from a European pied flycatcher (Ficedula hypoleuca). Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora serinuse n. sp. after its host, the domestic canary (S. canaria forma domestica).

Isospora streperae n. sp. (Apicomplexa: Eimeriidae) from a grey currawong (Strepera versicolour plumbea) (Passeriformes: Artamidae) in Western Australia.

A new species, Isospora streperae n. sp., (Apicomplexa: Eimeriidae) is described from a single grey currawong bird (Strepera versicolour) (subspecies S. v. plumbea) in Western Australia. Sporulated oocysts (n = 32) are spherical to subspherical, with smooth colourless bilayered oocyst wall, 1.0 µm thick (outer layer 0⋅8 µm, inner 0.2 µm thick). Oocyst with a polar granule, an oocyst residuum and two spheroidal to subspheroidal sporocysts. Oocyst length, 23.8 (20.4-25.0) µm; oocyst width, 22.5 (20.0-24.6) µm; a shape index of 1.06, with Stieda, substieda bodies. Micropyle is absent. Sporocysts with compressed sporocyst residuum and four sporozoites. Sporocyst length, 14.4 (12.5-15.2) µm; sporocyst width, 11.2 (10.6-14.0) µm, sporocyst L/W ratio, 1.29. Necropsy of the bird identified haemorrhaging along the ileum and jejunum, which is where Isospora oocysts were also mostly detected. Molecular analysis was conducted at three loci; the 18S, 28S ribosomal RNA and the mitochondrial cytochrome oxidase (COI) gene. At the 18S locus, I. streperae n. sp. exhibited 99.5% and 99.4% similarity respectively to an Isospora sp. (MS-2003) from a Southern cape sparrow (Passer melanurus melanurus) and Isospora dovati from a domestic pigeon (Columba livia domestica). At the 28S locus, I. streperae n. sp. exhibited 96.9% similarity to an Isospora sp. (MS-2003) from a grosbeak starling (Scissirostrum dubium) and 95.8% similarity with the Isospora sp. (MS-2003) from a Southern cape sparrow. At the COI locus, I. streperae n. sp. exhibited 95.0% similarity to Isospora sp. from a yellow-necked mouse (Apodemus flavicollis) from the Czech Republic. Based on morphological and molecular data, this isolate is a new species of Isospora, which is named Isospora streperae n. sp. after its host, the grey currawong (Strepera versicolour plumbea).

A new Caryospora coccidian species (Apicomplexa: Eimeriidae) from the laughing kookaburra (Dacelo novaeguineae).

A new Caryospora coccidian species is described from the laughing kookaburra (Dacelo novaeguineae). Sporulated oocysts (n=30) are ovoid in shape with a smooth, colourless, bilayered oocyst wall and measure 31.4×29.3 (30.0-32.0×28.0-31.0) µm with a shape index of 1.1. Oocysts contain one spheroidal to subspheroidal sporocyst, 21.2×20.6 (20.0-24.0×20.0-21.0) µm. A spheroidal shaped sporocyst residuum is present; micropyle, Stieda, substieda and parastieda bodies are absent. Vermiform sporozoites (n=8) are arranged either parallel or randomly in the sporocyst, measuring 17.0×4.8 (16.0-18.0×4.0-6.0) µm, with a L/W ratio of 3.5. There is a large spheroidal, posterior refractile body in the middle of the sporozoite. Morphologically, this new species is most similar to Caryospora. The prevalence of this parasite was 6.7% in birds sampled in the morning and 33.3% from those sampled after midday. Further molecular characterisation was conducted at two loci; the 18S and 28S ribosomal RNA (rRNA). At the 18S locus, the new species of Caryospora was most closely related to Besnoitia besnoiti (99.2% similarity) and Hammondia triffittae (98.8% similarity). Although, no 28S partial sequences from Caryospora were available in GenBank, the highest similarity was with B.besnoiti (91.3%). Based on morphological and molecular data, this coccidian parasite is a new species that to date has not been reported. The new coccidian parasite is named Caryospora daceloe n. sp. after its host D. novaeguineae (the laughing kookaburra).

Isospora anthochaerae n. sp. (Apicomplexa: Eimeriidae) from a Red wattlebird (Anthochaera carunculata) (Passeriformes: Meliphagidae) in Western Australia.

A new species, Isospora anthochaerae n. sp. is described from a Red wattlebird (Anthochaera carunculata). Sporulated oocysts (n=37) are subspherical, with smooth colourless to pale brown bilayered oocyst wall, 0.8 µm thick (outer layer 0·6 µm, inner 0.2 µm thick). Oocyst with 2 spheroidal to subspheroidal sporocysts. Oocyst length, 23.4 µm (20.0-26.0); oocyst width, 20.7 µm (19.0-22.0); oocyst length/width (L/W) ratio, 1.1. Micropyle, oocyst residuum and polar granule are absent. Sporocysts with compact sporocyst residuum and 4 sporozoites. Sporocyst length, 14.5 µm; sporocyst width, 10.1 µm sporocyst L/W ratio, 1.4. Molecular analysis was conducted at four loci; the ribosomal internal transcribed spacer (ITS), the 18S and 28S ribosomal RNA and the mitochondrial cytochrome oxidase gene (COI). At the COI locus, I. anthochaerae n. sp. exhibited 98.5% similarity to Isospora lesouefi from a Regent honeyeater (Xanthomyza phrygia) and 98% similarity with an Isospora sp. (iSAT5) from a blackcap (Sylvia atricapilla). Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in Red wattlebirds.

Morphological and molecular characterization of Eimeria paludosa coccidian parasite (Apicomplexa:Eimeriidae) in a dusky moorhen (Gallinula tenebrosa, Gould, 1846) in Australia.

An Eimeria species is described from a dusky moorhen (Gallinula tenebrosa). Sporulated oocysts (n = 40) are ovoid, with a pitted single-layered oocyst wall in young oocysts and a relatively smooth wall in the mature oocysts. Oocyst wall was 1.0 µm thick, oocysts measured 17.3 × 13.3 (16.3-17.9 × 12.7-13.9) µm, oocyst length/width (L/W) ratio, 1.3. Oocyst residuum was absent. A large polar granule was always observed in the centre of the micropyle and many small polar granules were observed when the focus was on the wall. Sporocysts are elongate-ovoid, 8.4 × 5.1 (8.0-8.9 × 4.9-5.5) µm, sporocyst L/W ratio, 1.6 (1.5-1.8), sporocyst residuum was present, composed of numerous granules in a spherical or ovoid mass. Each sporocyst contained 2 elongate sporozoites, 7.7 × 2.6 (7-10 × 2.2-3) µm. A spherical-ellipsoid posterior refractile body was found in the sporozoites. A nucleus is located immediately anterior to the posterior refractile body. When the oocyst measurements and features were compared with valid Eimeria species from hosts in the Rallidae family, this Eimeria species was identified as E. paludosa. This is the first report of E. paludosa in Australia and the dusky moorhen (Gallinula tenebrosa) in a new host for this species. Molecular analysis was conducted at three loci; the 18S and 28S ribosomal RNA genes and the mitochondrial cytochrome oxidase gene (COI). At the 18 S locus, E. paludosa shared 97.3% genetic similarity with Eimeria gruis (GenBank accession number: AB544336). It also shared 99.2% genetic similarity with Eimeria crecis (GenBank accession numbers: HE653904 and HE653905) and 98.5% similarity with Eimeria nenei (GenBank accession numbers: HE653906), both of which were identified from a corncrake (Crex crex) in the United Kingdom. At the 28S locus, E. paludosa shared 91.4% similarity with E. papillata from a chicken (Gallus gallus) in the USA. At COI locus, E. paludosa was in a clade by itself and shared 87.2% similarity with E. irresidua, from a European rabbit (Oryctolagus cuniculus) from the Czech Republic. This is the first molecular characterization of E. paludosa.

Identification and genetic characterization of a novel species of Choleoeimeria Schneider, 1875 from a captive-bred bilby (Thylacomyidae; Macrotis lagotis) (Reid, 1837) in Western Australia.

A novel Eimeria sp. from a captive-bred bilby (Macrotis lagotis Reid, 1837) has been identified in Western Australia. The bilby was bred at the Kanyana Wildlife Rehabilitation Centre, Perth, as part of the National Bilby Recovery Plan. Oocysts (n = 31) irregular blunt ellipsoidal, 17-18 × 11-12 (17.2 × 11.3); length/width (L/W) ratio 1.4-1.5 (1.5). Wall bi-layered, 0.8-1.0 (0.9) thick, outer layer smooth, c.2/3 of total thickness. Micropyle barely discernible. Oocyst residuum is absent, but 2-3 small polar granules are present. Sporocysts (n = 31) ovoidal, 7-8 × 5-6 (7.8 × 5.7); L/W ratio 1.3-1.4 (1.4). Stieda, sub-Stieda and para-Stieda bodies absent or indiscernible; sporocyst residuum present, usually as an irregular body consisting of numerous granules that appear to be membrane-bound or sometimes diffuse among sporozoites. Sporozoites vermiform with a robust refractile body. Further molecular characterization was conducted on the sporulated oocysts. At the 18S locus, it sat in a large clade of the phylogenetic tree with two isolates of Eimeria angustus from quendas (Isoodon obesulus Shaw, 1797) and the Choleoeimeria spp. It shared the highest identity with E. angustus (KU248093) at 98.84%; at the COI gene locus, it was unique and most closely related to Choleoeimeria taggarti, which is hosted by another species of marsupial, the yellow-footed antechinus (Antechinus flavipes flavipes), with 90.58% genetic similarity. Based on morphological and molecular data, this isolate is a new species and named as Choleoeimeria yangi n. sp.

Eimeria tiliquae n. sp. (Apicomplexa: Eimeriidae) from the shingleback skink (Tiliqua rugosa rugosa).

A new species, Eimeria tiliquae n. sp. is described from a shingleback skink (Tiliqua rugosa rugosa). Sporulated oocysts (n=50) are spherical to subspherical, with colourless trilaminate oocyst wall, 0.7±0.1 (0.5-0.75) thick. Oocyst with 4 spheroidal to subspheroidal sporocysts. Oocyst length, 13.7±0.9 (12.0-16.3); oocyst width, 12.8±0.9 (11.5-15.0); oocyst length/width (L/W) ratio, 1.07±0.05 (1.0-1.2). Micropyle, oocyst residuum and polar granule absent. Sporocysts with globular sporocyst residuum and 2 sporozoites. Sporocyst length, 6.0±0.6 (5.0-7.5); sporocyst width, 5.4±0.6 (4.0-7.0); sporocyst L/W ratio, 1.11±0.11 (1.0-1.5). Stieda, parastieda and substieda bodies absent. Phylogenetic analysis of 18S rRNA sequences indicated that E. tiliquae n. sp. shared 96.4-96.5% genetic similarity to E. tropidura, its closest relative. Reptile-derived sequences were not available for the mitochondrial cytochrome oxidase gene (COI) and phylogenetic analysis at this locus placed E. tiliquae n. sp. in a clade by itself but grouping closest (92% similarity) with a novel isolate from a King's skink (Egernia kingii) from Western Australia. Based on morphological and molecular data, this isolate is a new species of coccidian parasite that to date has only been found in shingleback skinks.