- Email: [email protected]
Prevalence of Cryptosporidium and Giardia species in animals in irrigation catchments in the southwest of Australia
Available online at www.sciencedirect.com
Experimental Parasitology 118 (2008) 596–599 www.elsevier.com/locate/yexpr
Research brief
Prevalence of Cryptosporidium and Giardia species in animals in irrigation catchments in the southwest of Australia Suzi McCarthy a, Josephine Ng a, Cameron Gordon b, Rachael Miller b, Andrew Wyber b, Una M. Ryan a,* a
Division of Health Sciences, School of Veterinary and Biomedical Science, Murdoch University, Murdoch, WA 6150, Australia b Water Corporation, Level 3, 629 Newcastle Street, Leederville, WA 6007, Australia Received 7 August 2007; received in revised form 18 October 2007; accepted 30 October 2007 Available online 5 November 2007
Abstract Screening of 445 animal faecal samples in irrigation catchments in Western Australia (WA) was conducted to identify the prevalence of Cryptosporidium and Giardia species. Of the samples positive for Giardia duodenalis, 30.7% (12/36) were the zoonotic Assemblage A, while approximately 13% (4/30) of Cryptosporidium positives were zoonotic. This is the first finding of Giardia Assemblage A in native marsupials and birds and indicates that marsupials and possibly birds may potentially be a reservoir of zoonotic Giardia. Ó 2007 Elsevier Inc. All rights reserved. Index Descriptors and Abbreviations: Cryptosporidium; Giardia; Catchments; Genotyping; Zoonotic; Marsupial; Environment
1. Introduction Cryptosporidium and Giardia are the most common causes of waterborne outbreaks of gastroenteritis worldwide (Pond, 2005). Cryptosporidium oocysts and Giardia cysts have been detected in pristine surface water, ground water, filtered drinking water, lakes, swimming pools and coastal marine waters (Hlavsa et al., 2005a,b; Kistemann et al., 2002). These parasites have gained the attention of water utilities world-wide due to drinking water related outbreaks, because the oocysts and cysts produced by these parasites are extremely hardy, easily spread via water, and difficult to inactivate or remove from water intended for consumption without the use of filtration (Chen et al., 2002; Fayer, 2004; Fayer et al., 2000). Although, Australia has yet to experience an outbreak, the Sydney water crisis in 1998, where high levels of (oo)cysts were found in the Warragamba catchment (Ryan et al., 2005a), highlighted the importance of identifying sources of zoonotic species *
Corresponding author. Fax: +61 89310 414. E-mail address: [email protected] (U.M. Ryan).
0014-4894/$ - see front matter Ó 2007 Elsevier Inc. All rights reserved. doi:10.1016/j.exppara.2007.10.014
in catchments. To date no genotyping studies have been undertaken to assess the potential risk of Cryptosporidium sp. and Giardia sp. contamination in Western Australia catchments. The present study was an attempt to understand the public health significance of these parasites in animals in southwestern Australian irrigation catchments using molecular tools. 2. Materials and methods A total of 445 animal faecal samples were collected directly from the ground from various wildlife and livestock species residing within three irrigation catchments; Waroona, a non-agricultural catchment consisting of state forest that has some recreational activities; Drakesbrook Weir which is used for recreational and agricultural use and Wokalup Pipehead dam which is used for agricultural but not recreational activities. The identity of the animal host of the faecal sample was determined in most cases by visual identification of the host whilst excreting and with the aid of a scat and tracking manual (Triggs, 2004), where defecation was not witnessed.
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599
Faecal samples were stored at 4 °C until processing using a QIAmp stool kit (Qiagen, Hilden, Germany). A two-step nested PCR protocol was used to amplify the 18S rDNA gene of Cryptosporidium as previously described (Ryan et al., 2003). For Giardia, a primary PCR product of 292 bp was amplified from the 18S rDNA gene as previously described (Hopkins et al., 1997). For the secondary PCR, a fragment of 130 bp was amplified using internal primers described by Read et al. (2002). PCR products were purified using a freeze–squeeze method (Ng et al., 2006) and sequenced using a Big Dye version 3.1 Terminator Cycle Sequencing Kit (Applied Biosystems, Foster City, California). Nucleotide sequences were analysed using Chromas v2.3 (http://www.technelysium.com.au) and aligned using Clustal W (Thompson et al., 1994). Statistical analysis was performed using SPSS 11.0 (Statistical Package for the Social Sciences) for Macintosh OS X (SPSS Inc., Chicago, USA) to determine if there was any association between the prevalence of Giardia and Cryptosporidium in each catchment and factors such as faecal consistency, seasonality, rainfall and temperature. Total daily rainfall and maximum daily temperatures were obtained from remote weather monitoring stations closet to the sampling site from the Bureau of Meteorology (www.bom.gov.au) for a period of 2 weeks prior to the sampling dates from each site. From this, the average temperature and rainfall was calculated, resulting in one average temperature and rainfall measurement for each sampling date. 3. Results and discussion Thirty Cryptosporidium positive isolates were detected by PCR, for a total prevalence of 6.7% (30 of 445) for the three irrigation catchments. A higher prevalence of Cryptosporidium (11.3%; 17 of 150) was detected at Waroona, than at the agricultural catchments (Drakesbrook Weir and Wokalup pipehead) 4.4% (13 of 295) (p-value 0.008). Sequences were obtained for 14 positives (see Table 1), the remaining 16 positives were unable to be sequenced due to insufficient template and/or mixed template. The non-zoonotic marsupial genotype I was identified in faeces from two kangaroos which supports an earlier study conducted by Power et al. (2005), on eastern grey kangaroos in a Sydney watershed. The Cryptosporidium cervine genotype was identified in faeces from an unknown host from Drakesbrook and the Cryptosporidium pig II genotype was identified in faeces from two pigs from Wokalup. Four Cryptosporidium parvum isolates were detected; two in faeces from beef cattle from Drakesbrook, one in faeces from a dairy cow from Wokalup and one in faeces from an unknown host from Waroona. Four Cryptosporidium bovis isolates were detected in faeces from three beef cattle from Drakesbrook and Wokalup and one in faeces from dairy cattle from Wokalup. Previous studies have detected C. bovis in cattle in the United States (Fayer et al., 2005) and sheep in Australia (Ryan et al., 2005b). This is the first report of C. bovis in cattle in Australia.
597
Cryptosporidium hominis was detected in a faecal sample from a wild bird from Waroona. This species has been previously reported in geese in Ohio and Illinois in the United States (Zhou et al., 2004) and was thought to be the result of mechanical transmission rather than an established infection and may also be the result of mechanical transmission in the present study. The presence of C. hominis in the present study could be attributed to the high recreational use, overcrowding and limited toilet facilities within the Waroona catchment. The overall prevalence of Giardia was 8.7% (39 of 445) and was detected in significantly higher (p-value 0.004) levels (11.5%; 34 of 295) in the agricultural catchments compared to Waroona at 3.3% (5 of 150). Sequences were obtained for 36/39 positives and of these, 61% (24/36) were the non-zoonotic G. duodenalis Assemblage E identified in sheep and cattle, and 30.7% (12 of 36) were the zoonotic Assemblage A identified in cattle, birds, kangaroos and a fox (Table 1). This study is the first report of Assemblage A in kangaroos. A previous study identified a novel hostadapted non-zoonotic marsupial Giardia genotype in one marsupial, Quenda (Isoodon obesulus) (Adams et al., 2004). In the present study, a total of 72 kangaroo isolates were screened but only three Assemblage A isolates were identified. Another study reported a prevalence of 61.5% (16 of 26) for Giardia in Eastern barred bandicoots in Tasmania, but did not conduct genotyping (Bettiol et al., 1997). The same study also reported that bandicoots could be experimentally infected with G. duodenalis from a human source, raising concern that wildlife may play a role in zoonotic transmission of Giardia. The finding of Assemblage A in kangaroos in the present study provides supporting evidence that marsupials may be a reservoir of zoonotic Giardia. The three kangaroos that were positive for Assemblage A were from the Waroona dam, which has substantial recreational use. It is therefore possible that the kangaroos may have acquired this zoonotic genotype from vegetation, soil or by drinking irrigation water contaminated from human activity. As water samples were not tested during this study, the source of the human-infectious Assemblage A is difficult to fully determine. Assemblage A was also found in a fox (Drakesbrook) similar to the study by Hamnes et al. (2007), in which Assemblage A was found in both adult and juvenile Norwegian red foxes. The finding of Assemblage A in a wild bird (Waroona Dam) is a first, but may be due to mechanical transmission. A preliminary assessment of environmental risk factors that may increase water shed contamination with Cryptosporidium and Giardia sp. indicated that periods of heavy rainfall (>200 mm/month) and cooler temperatures (mean daily temperature <17 °C) (p-value 0.004) appeared to be risk factors for the increased prevalence of Giardia. A similar pattern for Cryptosporidium was noted, however, it was not statistically significant (p > 0.05). The present study has been the first opportunity in Western Australia to assess the potential public health signifi-
598
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599
Table 1 Species and genotypes of Cryptosporidium and Giardia identified during this study Species/genotype
Host
Catchment
C. parvum C. parvum C. parvum C. hominis Cryptosporidium marsupial genotype I Cryptosporidium cervid genotype C. bovis C. bovis Cryptosporidium pig genotype II G. duodenalis Assemblage E Assemblage E Assemblage E G. duodenalis Assemblage A Assemblage A Assemblage A Assemblage A Assemblage A Assemblage A
Beef cattle (n = 2) Unknown (n = 1) Dairy cattle (n = 1) Wild bird (n = 1) Kangaroo (n = 2) Unknown (n = 1) Beef cattle (n = 3) Dairy cattle (n = 1) Pig (n = 2) Sheep (n = 3) Beef cattle (n = 17) Dairy cattle (n = 4) Dairy cow (n = 1) Beef cattle (n = 5) Fox (n = 1) Kangaroo (n = 3) Wild bird (n = 1) Unknown (n = 1)
Drakesbrook Waroona Wokalup Waroona Waroona Drakesbrook Drakesbrook Wokalup Wokalup Drakesbrook Drakesbrook Wokalup Wokalup Drakesbrook Drakesbrook Waroona Waroona Waroona
cance of faecal contamination from animals occupying water catchments using molecular tools. Although zoonotic Cryptosporidium and Giardia genotypes were detected at all three irrigation catchments, water sampling would be required to determine the likelihood and degree of contamination of the water from the animals. The degree of water contamination has public health implications for recreational users of the dams, water used for crop irrigation and economic implications for farms located downstream from contamination sites. The presence of zoonotic species of Cryptosporidium and Giardia warrants a more detailed examination with a larger number of host species and water samples. Acknowledgments This study was supported by the Water Corporation, WA. We thank the WA College of Agriculture, Harvey and the landowners within the Waroona, Drakesbrook and Wokalup catchments for their participation in this study. References Adams, P., Monis, P., Elliot, A., Thompson, R., 2004. Cyst morphology and sequence analysis of the small subunit rDNA and ef1 alpha identifies a novel Giardia genotype in a quenda (Isoodon obesulus) from Western Australia. Infection, Genetics and Evolution 4, 365– 367. Bettiol, S., Kettlewell, J., Davies, N., Goldsmid, J., 1997. Giardiasis in native marsupials of Tasmania. Journal of Wildlife Diseases 33, 352– 354. Chen, X.-M., Keithly, J.S., Paya, C.V., LaRusso, N.F., 2002. Cryptosporidiosis. New England Journal of Medicine 346, 1723–1731. Fayer, R., 2004. Cryptosporidium: a water-borne zoonotic parasite. Veterinary Parasitology 126, 37–56. Fayer, R., Morgan, U., Upton, S.J., 2000. Epidemiology of Cryptosporidium: transmission, detection and identification. International Journal for Parasitology 30, 1305–1322.
and Wokalup
and Wokalup and Wokalup
and Wokalup
Fayer, R., Santin, M., Xiao, L., 2005. Cryptosporidium bovis n. sp. (Apicomplexa: Cryptosporidiiae) in cattle (Bos taurus). Journal of Parasitology 91, 624–629. Hamnes, I., Gjerde, B., Forberg, T., Robertson, L., 2007. Occurrence of Giardia and Cryptosporidium in Norwegian red foxes (Vulpes vulpes). Veterinary Parasitology 143, 347–353. Hlavsa, M.C., Watson, J.C., Beach, M.J., 2005a. Cryptosporidiosis Surveillance—United States 1999–2002. Morbidity and Mortality Weekly Report: Surveillance Summaries 54, 1–8. Hlavsa, M.C., Watson, J.C., Beach, M.J., 2005b. Giardiasis Surveillance—United States, 1998–2002. Morbidity and Mortality Weekly Report: Surveillance Summaries 54, 9–16. Hopkins, R.M., Meloni, P., Groth, D., Wetherall, J.D., Reynoldson, J.A., Thompson, A.C.A., 1997. Ribosomal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality. Journal of Parasitology 83, 44–51. Kistemann, T., Classen, T., Koch, C., Dangendorf, F., Fischeder, R., Gebel, J., Vacata, V., Exner, M., 2002. Microbial load of drinking water reservoir tributaries during extreme rainfall and runoff. Applied and Environmental Microbiology 68, 2188–2197. Ng, J., Pavlasek, I., Ryan, U.M., 2006. Identification of novel Cryptosporidium genotypes from avian hosts. Applied and Environmental Microbiology 72, 7548–7553. Pond, K., 2005. Protozoa and trematodes. In: Organization, W.H. (Ed.), Water Recreation and Disease. Plausibility of Associated Infections: Acute Effects, Sequelae and Mortality. IWA Publishing, London, pp. 147–190. Power, M.L., Sangster, N.C., Slade, M.B., Veal, D.A., 2005. Patterns of Cryptosporidium oocyst shedding by Eastern Grey kangaroos inhabiting an Australian Watershed. Applied and Environmental Microbiology 71, 6159–6164. Read, C., Walters, J., Robertson, I.D., Thompson, A.C.A., 2002. Correlation between genotype of Giardia duodenalis and diarrhoea. International Journal for Parasitology 32, 229–231. Ryan, U., Xiao, L., Read, C., Zhou, L., Lal, A.A., Pavlasek, I., 2003. Identification of novel Cryptosporidium genotypes from the Czech Republic. Applied and Environmental Microbiology 69, 4302–4307. Ryan, U.M., Bath, C., Robertson, I., Read, C., Elliot, A., McInnes, L., Traub, R., Besier, B., 2005a. Sheep may not be an important zoonotic reservoir for Cryptosporidium and Giardia parasites. Applied and Environmental Microbiology 71, 4992–4997. Ryan, U.M., Read, C., Hawkins, P., Warnecke, M., Swanson, P., Griffith, M., Deere, D., Cunningham, M., Cox, P., 2005b. Genotypes of
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599 Cryptosporidium from Sydney water catchment areas. Journal of Applied Microbiology 98, 1221–1229. Thompson, J.D., Higgins, D.G., Gibson, T.J., 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673–4680.
599
Triggs, B., 2004. Tracks Scats and Other Traces: A Field Guide to Australian Mammals. Oxford University Press, South Melbourne. Zhou, L., Kassa, H., Tischler, M.L., Xiao, L., 2004. Host-adapted Cryptosporidium spp. in Canada geese (Branta canadensis). Applied and Environmental Microbiology 70, 4211–4215.
Experimental Parasitology 118 (2008) 596–599 www.elsevier.com/locate/yexpr
Research brief
Prevalence of Cryptosporidium and Giardia species in animals in irrigation catchments in the southwest of Australia Suzi McCarthy a, Josephine Ng a, Cameron Gordon b, Rachael Miller b, Andrew Wyber b, Una M. Ryan a,* a
Division of Health Sciences, School of Veterinary and Biomedical Science, Murdoch University, Murdoch, WA 6150, Australia b Water Corporation, Level 3, 629 Newcastle Street, Leederville, WA 6007, Australia Received 7 August 2007; received in revised form 18 October 2007; accepted 30 October 2007 Available online 5 November 2007
Abstract Screening of 445 animal faecal samples in irrigation catchments in Western Australia (WA) was conducted to identify the prevalence of Cryptosporidium and Giardia species. Of the samples positive for Giardia duodenalis, 30.7% (12/36) were the zoonotic Assemblage A, while approximately 13% (4/30) of Cryptosporidium positives were zoonotic. This is the first finding of Giardia Assemblage A in native marsupials and birds and indicates that marsupials and possibly birds may potentially be a reservoir of zoonotic Giardia. Ó 2007 Elsevier Inc. All rights reserved. Index Descriptors and Abbreviations: Cryptosporidium; Giardia; Catchments; Genotyping; Zoonotic; Marsupial; Environment
1. Introduction Cryptosporidium and Giardia are the most common causes of waterborne outbreaks of gastroenteritis worldwide (Pond, 2005). Cryptosporidium oocysts and Giardia cysts have been detected in pristine surface water, ground water, filtered drinking water, lakes, swimming pools and coastal marine waters (Hlavsa et al., 2005a,b; Kistemann et al., 2002). These parasites have gained the attention of water utilities world-wide due to drinking water related outbreaks, because the oocysts and cysts produced by these parasites are extremely hardy, easily spread via water, and difficult to inactivate or remove from water intended for consumption without the use of filtration (Chen et al., 2002; Fayer, 2004; Fayer et al., 2000). Although, Australia has yet to experience an outbreak, the Sydney water crisis in 1998, where high levels of (oo)cysts were found in the Warragamba catchment (Ryan et al., 2005a), highlighted the importance of identifying sources of zoonotic species *
Corresponding author. Fax: +61 89310 414. E-mail address: [email protected] (U.M. Ryan).
0014-4894/$ - see front matter Ó 2007 Elsevier Inc. All rights reserved. doi:10.1016/j.exppara.2007.10.014
in catchments. To date no genotyping studies have been undertaken to assess the potential risk of Cryptosporidium sp. and Giardia sp. contamination in Western Australia catchments. The present study was an attempt to understand the public health significance of these parasites in animals in southwestern Australian irrigation catchments using molecular tools. 2. Materials and methods A total of 445 animal faecal samples were collected directly from the ground from various wildlife and livestock species residing within three irrigation catchments; Waroona, a non-agricultural catchment consisting of state forest that has some recreational activities; Drakesbrook Weir which is used for recreational and agricultural use and Wokalup Pipehead dam which is used for agricultural but not recreational activities. The identity of the animal host of the faecal sample was determined in most cases by visual identification of the host whilst excreting and with the aid of a scat and tracking manual (Triggs, 2004), where defecation was not witnessed.
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599
Faecal samples were stored at 4 °C until processing using a QIAmp stool kit (Qiagen, Hilden, Germany). A two-step nested PCR protocol was used to amplify the 18S rDNA gene of Cryptosporidium as previously described (Ryan et al., 2003). For Giardia, a primary PCR product of 292 bp was amplified from the 18S rDNA gene as previously described (Hopkins et al., 1997). For the secondary PCR, a fragment of 130 bp was amplified using internal primers described by Read et al. (2002). PCR products were purified using a freeze–squeeze method (Ng et al., 2006) and sequenced using a Big Dye version 3.1 Terminator Cycle Sequencing Kit (Applied Biosystems, Foster City, California). Nucleotide sequences were analysed using Chromas v2.3 (http://www.technelysium.com.au) and aligned using Clustal W (Thompson et al., 1994). Statistical analysis was performed using SPSS 11.0 (Statistical Package for the Social Sciences) for Macintosh OS X (SPSS Inc., Chicago, USA) to determine if there was any association between the prevalence of Giardia and Cryptosporidium in each catchment and factors such as faecal consistency, seasonality, rainfall and temperature. Total daily rainfall and maximum daily temperatures were obtained from remote weather monitoring stations closet to the sampling site from the Bureau of Meteorology (www.bom.gov.au) for a period of 2 weeks prior to the sampling dates from each site. From this, the average temperature and rainfall was calculated, resulting in one average temperature and rainfall measurement for each sampling date. 3. Results and discussion Thirty Cryptosporidium positive isolates were detected by PCR, for a total prevalence of 6.7% (30 of 445) for the three irrigation catchments. A higher prevalence of Cryptosporidium (11.3%; 17 of 150) was detected at Waroona, than at the agricultural catchments (Drakesbrook Weir and Wokalup pipehead) 4.4% (13 of 295) (p-value 0.008). Sequences were obtained for 14 positives (see Table 1), the remaining 16 positives were unable to be sequenced due to insufficient template and/or mixed template. The non-zoonotic marsupial genotype I was identified in faeces from two kangaroos which supports an earlier study conducted by Power et al. (2005), on eastern grey kangaroos in a Sydney watershed. The Cryptosporidium cervine genotype was identified in faeces from an unknown host from Drakesbrook and the Cryptosporidium pig II genotype was identified in faeces from two pigs from Wokalup. Four Cryptosporidium parvum isolates were detected; two in faeces from beef cattle from Drakesbrook, one in faeces from a dairy cow from Wokalup and one in faeces from an unknown host from Waroona. Four Cryptosporidium bovis isolates were detected in faeces from three beef cattle from Drakesbrook and Wokalup and one in faeces from dairy cattle from Wokalup. Previous studies have detected C. bovis in cattle in the United States (Fayer et al., 2005) and sheep in Australia (Ryan et al., 2005b). This is the first report of C. bovis in cattle in Australia.
597
Cryptosporidium hominis was detected in a faecal sample from a wild bird from Waroona. This species has been previously reported in geese in Ohio and Illinois in the United States (Zhou et al., 2004) and was thought to be the result of mechanical transmission rather than an established infection and may also be the result of mechanical transmission in the present study. The presence of C. hominis in the present study could be attributed to the high recreational use, overcrowding and limited toilet facilities within the Waroona catchment. The overall prevalence of Giardia was 8.7% (39 of 445) and was detected in significantly higher (p-value 0.004) levels (11.5%; 34 of 295) in the agricultural catchments compared to Waroona at 3.3% (5 of 150). Sequences were obtained for 36/39 positives and of these, 61% (24/36) were the non-zoonotic G. duodenalis Assemblage E identified in sheep and cattle, and 30.7% (12 of 36) were the zoonotic Assemblage A identified in cattle, birds, kangaroos and a fox (Table 1). This study is the first report of Assemblage A in kangaroos. A previous study identified a novel hostadapted non-zoonotic marsupial Giardia genotype in one marsupial, Quenda (Isoodon obesulus) (Adams et al., 2004). In the present study, a total of 72 kangaroo isolates were screened but only three Assemblage A isolates were identified. Another study reported a prevalence of 61.5% (16 of 26) for Giardia in Eastern barred bandicoots in Tasmania, but did not conduct genotyping (Bettiol et al., 1997). The same study also reported that bandicoots could be experimentally infected with G. duodenalis from a human source, raising concern that wildlife may play a role in zoonotic transmission of Giardia. The finding of Assemblage A in kangaroos in the present study provides supporting evidence that marsupials may be a reservoir of zoonotic Giardia. The three kangaroos that were positive for Assemblage A were from the Waroona dam, which has substantial recreational use. It is therefore possible that the kangaroos may have acquired this zoonotic genotype from vegetation, soil or by drinking irrigation water contaminated from human activity. As water samples were not tested during this study, the source of the human-infectious Assemblage A is difficult to fully determine. Assemblage A was also found in a fox (Drakesbrook) similar to the study by Hamnes et al. (2007), in which Assemblage A was found in both adult and juvenile Norwegian red foxes. The finding of Assemblage A in a wild bird (Waroona Dam) is a first, but may be due to mechanical transmission. A preliminary assessment of environmental risk factors that may increase water shed contamination with Cryptosporidium and Giardia sp. indicated that periods of heavy rainfall (>200 mm/month) and cooler temperatures (mean daily temperature <17 °C) (p-value 0.004) appeared to be risk factors for the increased prevalence of Giardia. A similar pattern for Cryptosporidium was noted, however, it was not statistically significant (p > 0.05). The present study has been the first opportunity in Western Australia to assess the potential public health signifi-
598
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599
Table 1 Species and genotypes of Cryptosporidium and Giardia identified during this study Species/genotype
Host
Catchment
C. parvum C. parvum C. parvum C. hominis Cryptosporidium marsupial genotype I Cryptosporidium cervid genotype C. bovis C. bovis Cryptosporidium pig genotype II G. duodenalis Assemblage E Assemblage E Assemblage E G. duodenalis Assemblage A Assemblage A Assemblage A Assemblage A Assemblage A Assemblage A
Beef cattle (n = 2) Unknown (n = 1) Dairy cattle (n = 1) Wild bird (n = 1) Kangaroo (n = 2) Unknown (n = 1) Beef cattle (n = 3) Dairy cattle (n = 1) Pig (n = 2) Sheep (n = 3) Beef cattle (n = 17) Dairy cattle (n = 4) Dairy cow (n = 1) Beef cattle (n = 5) Fox (n = 1) Kangaroo (n = 3) Wild bird (n = 1) Unknown (n = 1)
Drakesbrook Waroona Wokalup Waroona Waroona Drakesbrook Drakesbrook Wokalup Wokalup Drakesbrook Drakesbrook Wokalup Wokalup Drakesbrook Drakesbrook Waroona Waroona Waroona
cance of faecal contamination from animals occupying water catchments using molecular tools. Although zoonotic Cryptosporidium and Giardia genotypes were detected at all three irrigation catchments, water sampling would be required to determine the likelihood and degree of contamination of the water from the animals. The degree of water contamination has public health implications for recreational users of the dams, water used for crop irrigation and economic implications for farms located downstream from contamination sites. The presence of zoonotic species of Cryptosporidium and Giardia warrants a more detailed examination with a larger number of host species and water samples. Acknowledgments This study was supported by the Water Corporation, WA. We thank the WA College of Agriculture, Harvey and the landowners within the Waroona, Drakesbrook and Wokalup catchments for their participation in this study. References Adams, P., Monis, P., Elliot, A., Thompson, R., 2004. Cyst morphology and sequence analysis of the small subunit rDNA and ef1 alpha identifies a novel Giardia genotype in a quenda (Isoodon obesulus) from Western Australia. Infection, Genetics and Evolution 4, 365– 367. Bettiol, S., Kettlewell, J., Davies, N., Goldsmid, J., 1997. Giardiasis in native marsupials of Tasmania. Journal of Wildlife Diseases 33, 352– 354. Chen, X.-M., Keithly, J.S., Paya, C.V., LaRusso, N.F., 2002. Cryptosporidiosis. New England Journal of Medicine 346, 1723–1731. Fayer, R., 2004. Cryptosporidium: a water-borne zoonotic parasite. Veterinary Parasitology 126, 37–56. Fayer, R., Morgan, U., Upton, S.J., 2000. Epidemiology of Cryptosporidium: transmission, detection and identification. International Journal for Parasitology 30, 1305–1322.
and Wokalup
and Wokalup and Wokalup
and Wokalup
Fayer, R., Santin, M., Xiao, L., 2005. Cryptosporidium bovis n. sp. (Apicomplexa: Cryptosporidiiae) in cattle (Bos taurus). Journal of Parasitology 91, 624–629. Hamnes, I., Gjerde, B., Forberg, T., Robertson, L., 2007. Occurrence of Giardia and Cryptosporidium in Norwegian red foxes (Vulpes vulpes). Veterinary Parasitology 143, 347–353. Hlavsa, M.C., Watson, J.C., Beach, M.J., 2005a. Cryptosporidiosis Surveillance—United States 1999–2002. Morbidity and Mortality Weekly Report: Surveillance Summaries 54, 1–8. Hlavsa, M.C., Watson, J.C., Beach, M.J., 2005b. Giardiasis Surveillance—United States, 1998–2002. Morbidity and Mortality Weekly Report: Surveillance Summaries 54, 9–16. Hopkins, R.M., Meloni, P., Groth, D., Wetherall, J.D., Reynoldson, J.A., Thompson, A.C.A., 1997. Ribosomal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality. Journal of Parasitology 83, 44–51. Kistemann, T., Classen, T., Koch, C., Dangendorf, F., Fischeder, R., Gebel, J., Vacata, V., Exner, M., 2002. Microbial load of drinking water reservoir tributaries during extreme rainfall and runoff. Applied and Environmental Microbiology 68, 2188–2197. Ng, J., Pavlasek, I., Ryan, U.M., 2006. Identification of novel Cryptosporidium genotypes from avian hosts. Applied and Environmental Microbiology 72, 7548–7553. Pond, K., 2005. Protozoa and trematodes. In: Organization, W.H. (Ed.), Water Recreation and Disease. Plausibility of Associated Infections: Acute Effects, Sequelae and Mortality. IWA Publishing, London, pp. 147–190. Power, M.L., Sangster, N.C., Slade, M.B., Veal, D.A., 2005. Patterns of Cryptosporidium oocyst shedding by Eastern Grey kangaroos inhabiting an Australian Watershed. Applied and Environmental Microbiology 71, 6159–6164. Read, C., Walters, J., Robertson, I.D., Thompson, A.C.A., 2002. Correlation between genotype of Giardia duodenalis and diarrhoea. International Journal for Parasitology 32, 229–231. Ryan, U., Xiao, L., Read, C., Zhou, L., Lal, A.A., Pavlasek, I., 2003. Identification of novel Cryptosporidium genotypes from the Czech Republic. Applied and Environmental Microbiology 69, 4302–4307. Ryan, U.M., Bath, C., Robertson, I., Read, C., Elliot, A., McInnes, L., Traub, R., Besier, B., 2005a. Sheep may not be an important zoonotic reservoir for Cryptosporidium and Giardia parasites. Applied and Environmental Microbiology 71, 4992–4997. Ryan, U.M., Read, C., Hawkins, P., Warnecke, M., Swanson, P., Griffith, M., Deere, D., Cunningham, M., Cox, P., 2005b. Genotypes of
S. McCarthy et al. / Experimental Parasitology 118 (2008) 596–599 Cryptosporidium from Sydney water catchment areas. Journal of Applied Microbiology 98, 1221–1229. Thompson, J.D., Higgins, D.G., Gibson, T.J., 1994. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673–4680.
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