Una Ryan | Murdoch University (original) (raw)
Papers by Una Ryan
Experimental Parasitology, 2014
Journal of Tissue Culture Methods, Feb 28, 1986
Jex a R Ryan U M Ng J Campbell B E Xiao L Stevens M and Gasser R B Specific and Genotypic Identification of Cryptosporidium from a Broad Range of Host Species By Nonisotopic Sscp Analysis of Nuclear Ribosomal Dna Electrophoresis 28 Pp 2818 2825, Aug 1, 2007
The accurate identification of Cryptosporidium (Protozoa: Apicomplexa) species and genotypes is c... more The accurate identification of Cryptosporidium (Protozoa: Apicomplexa) species and genotypes is central to the understanding of the transmission and to the diagnosis and control of cryptosporidiosis. In this study, we demonstrate the effectiveness of nonisotopic SSCP analysis of a approximately 300 bp region of the small subunit (pSSU) of ribosomal DNA for the specific identification of and delineation among 18 different Cryptosporidium species and genotypes from a wide range of hosts. This mutation scanning approach allowed the rapid and reliable differentiation between species/genotypes differing by as little as 1.3% in the pSSU sequence, with the capacity to detect point mutations. The present findings confirm the usefulness of this tool for the rapid genetic screening of Cryptosporidium samples from any host species, providing a foundation for detailed systematic, epidemiological and ecological studies. Although applied herein to pSSU, this low cost approach should be applicable to a wide range of genetic loci for population genetic investigations of Cryptosporidium.
O Brien E Mcinnes L and Ryan U Cryptosporidium Gp60 Genotypes from Humans and Domesticated Animals in Australia North America and Europe Experimental Parasitology 118 Pp 118 121, 2008
In Vitro Cell Dev Biol Animal, 1997
Yang R Ying J L J Monis P and Ryan U Molecular Characterisation of Cryptosporidium and Giardia in Cats in Western Australia Experimental Parasitology 155 Pp 13 18, May 7, 2015
Fitzgerald L Bennett M D Ng J Nicholls P K James F E Elliot a Slaven M and Ryan U Morphological and Molecular Characterisation of a Mixed Cryptosporidium Muris Cryptosporidium Felis Infection in a Cat Veterinary Parasitology 175 Pp 160 164, Jan 10, 2011
Sweeny J P a Robertson I D Ryan U Jacobson C and Woodgate R G Impacts of Naturally Acquired Protozoa and Strongylid Nematode Infections on Growth and Faecal Attributes in Lambs Veterinary Parasitology 184 Pp 298 308, Mar 23, 2012
Sweeny J P a Ryan U M Robertson I D and Jacobson C Cryptosporidium and Giardia Associated With Reduced Lamb Carcase Productivity Veterinary Parasitology 182 Pp 127 139, Jun 12, 2011
Ryan U M and Xiao L Molecular Epidemiology and Typing of Non Human Isolates of Cryptosporidium in Ortega Pierres G Caccio S Fayer R Mank T G Smith H V and Thompson R C a Giardia and Cryptosporidium from Molecules to Disease Cabi Wallingford Oxfordshire Uk Pp 65 80, 2009
... C. scophthalmi in fish; C. wrairi from guinea pigs; C. felis in cats; and C. canis in dogs (F... more ... C. scophthalmi in fish; C. wrairi from guinea pigs; C. felis in cats; and C. canis in dogs (Fayer et al., 2000, 2001; Alvarez-Pellitero and Sitjà-Bobadilla, 2002; Morgan-Ryan et al., 2002; Ryan et al., 2003a, 2004a; Alvarez-Pellitero et al., Cryptosporidium parvum and C. hominis are ...
Environmental Health Perspectives, Apr 1, 2006
Johnson J Samarasinghe B Buddle R Armson a and Ryan U Molecular Identification and Prevalence of Isospora Sp in Pigs in Western Australia Using a Pcr Rflp Assay Experimental Parasitology 120 Pp 191 193, Jul 1, 2008
Yang R Murphy C Song Y Ng Hublin J S Y Estcourt a Hijjawi N Chalmers R Hadfield S Bath a Gordon C and Ryan U Specific and Quantitative Detection and Identification of Cryptosporidium Hominis and C Parvum in Clinical and Environmental Samples Experimental Parasitology 135 Pp 142 147, Jul 6, 2013
Ryan U M Molecular Characterisation and Taxonomy of Cryptosporidium in Thompson R C a Armson a and Ryan U Cryptosporidium from Molecules to Disease Elsevier Amsterdam the Netherlands Pp 147 160, 2003
Yang R Paparini a Monis P and Ryan U M Comparison of Next Generation Droplet Digital Pcr With Quantitative Pcr For Enumeration of Cryptosporidium Oocysts in Combined Biological Sciences Meeting 29 August Perth Western Australia, Sep 13, 2014
Clinical microbiology laboratories rely on quantitative PCR for its speed, sensitivity, specifici... more Clinical microbiology laboratories rely on quantitative PCR for its speed, sensitivity, specificity and ease-of-use. However, quantitative PCR quantitation requires the use of a standard curve or normalisation to reference genes. Droplet digital PCR provides absolute quantitation without the need for calibration curves. A comparison between droplet digital PCR and quantitative PCR-based analyses was conducted for the enteric parasite Cryptosporidium, which is an important cause of gastritis in both humans and animals. Two loci were analysed (18S rRNA and actin) using a range of Cryptosporidium DNA templates, including recombinant plasmids, purified haemocytometer-counted oocysts, commercial flow cytometry-counted oocysts and faecal DNA samples from sheep, cattle and humans. Each method was evaluated for linearity, precision, limit of detection and cost. Across the same range of detection, both methods showed a high degree of linearity and positive correlation for standards (R(2)⩾0.999) and faecal samples (R(2)⩾0.9750). The precision of droplet digital PCR, as measured by mean Relative Standard Deviation (RSD;%), was consistently better compared with quantitative PCR, particularly for the 18S rRNA locus, but was poorer as DNA concentration decreased. The quantitative detection of quantitative PCR was unaffected by DNA concentration, but droplet digital PCR quantitative PCR was less affected by the presence of inhibitors, compared with quantitative PCR. For most templates analysed including Cryptosporidium-positive faecal DNA, the template copy numbers, as determined by droplet digital PCR, were consistently lower than by quantitative PCR. However, the quantitations obtained by quantitative PCR are dependent on the accuracy of the standard curve and when the quantitative PCR data were corrected for pipetting and DNA losses (as determined by droplet digital PCR), then the sensitivity of both methods was comparable. A cost analysis based on 96 samples revealed that the overall cost (consumables and labour) of droplet digital PCR was two times higher than quantitative PCR. Using droplet digital PCR to precisely quantify standard dilutions used for high-throughput and cost-effective amplifications by quantitative PCR would be one way to combine the advantages of the two technologies.
Experimental Parasitology, 2015
A new species, Isospora serinuse n. sp., (Apicomplexa:Eimeriidae) is described from a single dome... more 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).
Prostaglandins, 1975
ABSTRACT
Parasitology, 2015
Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, ... more Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro. Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.
Experimental Parasitology, 2014
Journal of Tissue Culture Methods, Feb 28, 1986
Jex a R Ryan U M Ng J Campbell B E Xiao L Stevens M and Gasser R B Specific and Genotypic Identification of Cryptosporidium from a Broad Range of Host Species By Nonisotopic Sscp Analysis of Nuclear Ribosomal Dna Electrophoresis 28 Pp 2818 2825, Aug 1, 2007
The accurate identification of Cryptosporidium (Protozoa: Apicomplexa) species and genotypes is c... more The accurate identification of Cryptosporidium (Protozoa: Apicomplexa) species and genotypes is central to the understanding of the transmission and to the diagnosis and control of cryptosporidiosis. In this study, we demonstrate the effectiveness of nonisotopic SSCP analysis of a approximately 300 bp region of the small subunit (pSSU) of ribosomal DNA for the specific identification of and delineation among 18 different Cryptosporidium species and genotypes from a wide range of hosts. This mutation scanning approach allowed the rapid and reliable differentiation between species/genotypes differing by as little as 1.3% in the pSSU sequence, with the capacity to detect point mutations. The present findings confirm the usefulness of this tool for the rapid genetic screening of Cryptosporidium samples from any host species, providing a foundation for detailed systematic, epidemiological and ecological studies. Although applied herein to pSSU, this low cost approach should be applicable to a wide range of genetic loci for population genetic investigations of Cryptosporidium.
O Brien E Mcinnes L and Ryan U Cryptosporidium Gp60 Genotypes from Humans and Domesticated Animals in Australia North America and Europe Experimental Parasitology 118 Pp 118 121, 2008
In Vitro Cell Dev Biol Animal, 1997
Yang R Ying J L J Monis P and Ryan U Molecular Characterisation of Cryptosporidium and Giardia in Cats in Western Australia Experimental Parasitology 155 Pp 13 18, May 7, 2015
Fitzgerald L Bennett M D Ng J Nicholls P K James F E Elliot a Slaven M and Ryan U Morphological and Molecular Characterisation of a Mixed Cryptosporidium Muris Cryptosporidium Felis Infection in a Cat Veterinary Parasitology 175 Pp 160 164, Jan 10, 2011
Sweeny J P a Robertson I D Ryan U Jacobson C and Woodgate R G Impacts of Naturally Acquired Protozoa and Strongylid Nematode Infections on Growth and Faecal Attributes in Lambs Veterinary Parasitology 184 Pp 298 308, Mar 23, 2012
Sweeny J P a Ryan U M Robertson I D and Jacobson C Cryptosporidium and Giardia Associated With Reduced Lamb Carcase Productivity Veterinary Parasitology 182 Pp 127 139, Jun 12, 2011
Ryan U M and Xiao L Molecular Epidemiology and Typing of Non Human Isolates of Cryptosporidium in Ortega Pierres G Caccio S Fayer R Mank T G Smith H V and Thompson R C a Giardia and Cryptosporidium from Molecules to Disease Cabi Wallingford Oxfordshire Uk Pp 65 80, 2009
... C. scophthalmi in fish; C. wrairi from guinea pigs; C. felis in cats; and C. canis in dogs (F... more ... C. scophthalmi in fish; C. wrairi from guinea pigs; C. felis in cats; and C. canis in dogs (Fayer et al., 2000, 2001; Alvarez-Pellitero and Sitjà-Bobadilla, 2002; Morgan-Ryan et al., 2002; Ryan et al., 2003a, 2004a; Alvarez-Pellitero et al., Cryptosporidium parvum and C. hominis are ...
Environmental Health Perspectives, Apr 1, 2006
Johnson J Samarasinghe B Buddle R Armson a and Ryan U Molecular Identification and Prevalence of Isospora Sp in Pigs in Western Australia Using a Pcr Rflp Assay Experimental Parasitology 120 Pp 191 193, Jul 1, 2008
Yang R Murphy C Song Y Ng Hublin J S Y Estcourt a Hijjawi N Chalmers R Hadfield S Bath a Gordon C and Ryan U Specific and Quantitative Detection and Identification of Cryptosporidium Hominis and C Parvum in Clinical and Environmental Samples Experimental Parasitology 135 Pp 142 147, Jul 6, 2013
Ryan U M Molecular Characterisation and Taxonomy of Cryptosporidium in Thompson R C a Armson a and Ryan U Cryptosporidium from Molecules to Disease Elsevier Amsterdam the Netherlands Pp 147 160, 2003
Yang R Paparini a Monis P and Ryan U M Comparison of Next Generation Droplet Digital Pcr With Quantitative Pcr For Enumeration of Cryptosporidium Oocysts in Combined Biological Sciences Meeting 29 August Perth Western Australia, Sep 13, 2014
Clinical microbiology laboratories rely on quantitative PCR for its speed, sensitivity, specifici... more Clinical microbiology laboratories rely on quantitative PCR for its speed, sensitivity, specificity and ease-of-use. However, quantitative PCR quantitation requires the use of a standard curve or normalisation to reference genes. Droplet digital PCR provides absolute quantitation without the need for calibration curves. A comparison between droplet digital PCR and quantitative PCR-based analyses was conducted for the enteric parasite Cryptosporidium, which is an important cause of gastritis in both humans and animals. Two loci were analysed (18S rRNA and actin) using a range of Cryptosporidium DNA templates, including recombinant plasmids, purified haemocytometer-counted oocysts, commercial flow cytometry-counted oocysts and faecal DNA samples from sheep, cattle and humans. Each method was evaluated for linearity, precision, limit of detection and cost. Across the same range of detection, both methods showed a high degree of linearity and positive correlation for standards (R(2)⩾0.999) and faecal samples (R(2)⩾0.9750). The precision of droplet digital PCR, as measured by mean Relative Standard Deviation (RSD;%), was consistently better compared with quantitative PCR, particularly for the 18S rRNA locus, but was poorer as DNA concentration decreased. The quantitative detection of quantitative PCR was unaffected by DNA concentration, but droplet digital PCR quantitative PCR was less affected by the presence of inhibitors, compared with quantitative PCR. For most templates analysed including Cryptosporidium-positive faecal DNA, the template copy numbers, as determined by droplet digital PCR, were consistently lower than by quantitative PCR. However, the quantitations obtained by quantitative PCR are dependent on the accuracy of the standard curve and when the quantitative PCR data were corrected for pipetting and DNA losses (as determined by droplet digital PCR), then the sensitivity of both methods was comparable. A cost analysis based on 96 samples revealed that the overall cost (consumables and labour) of droplet digital PCR was two times higher than quantitative PCR. Using droplet digital PCR to precisely quantify standard dilutions used for high-throughput and cost-effective amplifications by quantitative PCR would be one way to combine the advantages of the two technologies.
Experimental Parasitology, 2015
A new species, Isospora serinuse n. sp., (Apicomplexa:Eimeriidae) is described from a single dome... more 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).
Prostaglandins, 1975
ABSTRACT
Parasitology, 2015
Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, ... more Trypanosomes are blood-borne parasites that can cause severe disease in both humans and animals, yet little is known of the pathogenicity and life-cycles of trypanosomes in native Australian mammals. Trypanosoma copemani is known to be infective to a variety of Australian marsupials and has recently been shown to be potentially zoonotic as it is resistant to normal human serum. In the present study, in vivo and in vitro examination of blood and cultures from Australian marsupials was conducted using light microscopy, immunofluorescence, scanning electron microscopy and fluorescence in situ hybridization. Promastigote, sphaeromastigote and amastigote life-cycle stages were detected in vivo and in vitro. Novel trypanosome-like stages were also detected both in vivo and in vitro representing an oval stage, an extremely thin stage, an adherent stage and a tiny round stage. The tiny round and adherent stages appeared to adhere to erythrocytes causing potential haematological damage with clinical effects similar to haemolytic anaemia. The present study shows for the first time that trypomastigotes are not the only life-cycle stages circulating within the blood stream of trypanosome infected Australian native marsupials and provides insights into possible pathogenic mechanisms of this potentially zoonotic trypanosome species.