Genotypes of Cryptosporidium from Sydney water catchment areas (original) (raw)
Related papers
Meeting Report: Application of Genotyping Methods to Assess Risks from Cryptosporidium in Watersheds
Environmental Health Perspectives, 2005
A workshop titled "Application of Genotyping Methods to Assess Pathogen Risks from Cryptosporidium in Drinking Water Catchments" was held at the International Water Association biennial conference, Marrakech, Morocco, 23 September 2004. The workshop presented and discussed the findings of an interlaboratory trial that compared methods for genotyping Cryptosporidium oocysts isolated from feces. The primary goal of the trial and workshop was to assess the utility of current Cryptosporidium genotyping methods for determining the public health significance of oocysts isolated from feces in potable-water-supply watersheds. An expert panel of 16 watershed managers, public health practitioners, and molecular parasitologists was assembled for the workshop. A subordinate goal of the workshop was to educate watershed management and public health practitioners. An open invitation was extended to all conference delegates to attend the workshop, which drew approximately 50 interested delegates. In this report we summarize the peer consensus emerging from the workshop. Recommendations on the use of current methods by watershed managers and public health practitioners were proposed. Importantly, all the methods that were reported in the trial were mutually supporting and found to be valuable and worthy of further utility and development. Where there were choices as to which method to apply, the small-subunit ribosomal RNA gene was considered to be the optimum genetic locus to target. The single-strand conformational polymorphism method was considered potentially the most valuable for discriminating to the subtype level and where a large number of samples were to be analyzed. A research agenda for protozoan geneticists was proposed to improve the utility of methods into the future. Standardization of methods and nomenclature was promoted.
Applied and Environmental Microbiology, 2013
Nearly 690 raw surface water samples were collected during a 6-year period from multiple watersheds in the South Nation River basin, Ontario, Canada. Cryptosporidium oocysts in water samples were enumerated, sequenced, and genotyped by detailed phylogenetic analysis. The resulting species and genotypes were assigned to broad, known host and human infection risk classes. Wildlife/unknown, livestock, avian, and human host classes occurred in 21, 13, 3, and <1% of sampled surface waters, respectively. Cryptosporidium andersoni was the most commonly detected livestock species, while muskrat I and II genotypes were the most dominant wildlife genotypes. The presence of Giardia spp., Salmonella spp., Campylobacter spp., and Escherichia coli O157:H7 was evaluated in all water samples. The greatest significant odds ratios (odds of pathogen presence when host class is present/odds of pathogen presence when host class is absent) for Giardia spp., Campylobacter spp., and Salmonella spp. in water were associated, respectively, with livestock (odds ratio of 3.1), avian (4.
Cryptosporidiosis is one of the most common waterborne diseases reported worldwide. Outbreaks of this gastrointestinal disease, which is caused by the Cryptosporidium parasite, are often attributed to public swimming pools and municipal water supplies. Between the months of January and April in 2009, New South Wales, Australia, experienced the largest waterborne cryptosporidiosis outbreak reported in Australia to date. Through the course of the contamination event, 1,141 individuals became infected with Cryptosporidium. Health authorities in New South Wales indicated that public swimming pool use was a contributing factor in the outbreak. To identify the Cryptosporidium species responsible for the outbreak, fecal samples from infected patients were collected from hospitals and pathology companies throughout New South Wales for genetic analyses. Genetic characterization of Cryptosporidium oocysts from the fecal samples identified the anthroponotic Cryptosporidium hominis IbA10G2 subtype as the causative parasite. Equal proportions of infections were found in males and females, and an increased susceptibility was observed in the 0- to 4-year age group. Spatiotemporal analysis indicated that the outbreak was primarily confined to the densely populated coastal cities of Sydney and Newcastle.
Journal of Water and Health, 2018
Cryptosporidium, faecal indicator organisms and physical and chemical water quality variables were monitored in a small mixed rural–urban watershed in southeastern Brazil. Cryptosporidium was present in 43% of 117 water samples analysed by microscopy. Concentrations varied from non-detects to 14 oocysts L−1. All samples were further analysed by nested-PCR, and Cryptosporidium spp. were detected in 24% (28) of them. Sequencing at the 18S rRNA locus gave high quality sequences in eight samples, revealing the presence of Cryptosporidium parvum. Cryptosporidium was not correlated with faecal indicator organisms (total coliforms, Escherichia coli, Enterococcus and coliphages), nor with physical and chemical water quality variables (e.g. turbidity, electrical conductivity and chemical oxygen demand), but it was with farm animal density (number of animals per ha). Land use mapping reinforced the suggestions from Cryptosporidium genotyping that both animals (livestock) and humans are potent...
Water Supply, 2019
This study aimed to estimate the annual probability of Giardia and Cryptosporidium infection for a population supplied by contaminated drinking water sources. Parasites were quantified by the USEPA Method 1623.1/2012. Annual risk was estimated using the quantitative microbial risk assessment (QMRA) approach. Genotyping was performed using specific primers based on the 18S rRNA gene for Cryptosporidium and gdh gene for Giardia. Giardia was detected in 83.3% of the samples (<0.1 to 8.6 cysts/L) and Cryptosporidium in 37.5% (<0.1 to 2 oocysts/L). In general, annual risk values for Giardia were 1 log higher when compared with those obtained for Cryptosporidium. Giardia intestinalis A and B were present as well as C. hominis and C. parvum. The lack of protection measures for the water supply catchment point put the population's health at risk. The results provide data to support decision-makers to take actions to improve environmental quality resulting in a positive impact on c...
Electrophoresis, 2013
In the present study, we undertook a molecular epidemiological survey of Cryptosporidium and Giardia in calves on three dairy and two beef farms within an open drinking water catchment area (Melbourne, Australia). Faecal samples (n = 474) were collected from calves at two time points (5 months apart) and tested using a PCR-based mutation scanning-targeted sequencing phylogenetic approach, employing regions within the genes of small subunit (SSU) of ribosomal RNA (designated partial SSU), 60 kDa glycoprotein (pgp60) and triose phosphate isomerase (ptpi) as genetic markers. Using partial SSU, the C. bovis, C. parvum, C. ryanae and a new genotype of Cryptosporidium were characterised from totals of 74 (15.6%), 35 (7.3%), 37 (7.8%) and 9 (1.9%) samples, respectively. Using pgp60, C. parvum genotype IIa subgenotype A18G3R1 was detected in 29 samples. Using ptpi, G. duodenalis assemblages A and E were detected in totals of 10 (2.1%) and 130 (27.4%) samples, respectively. The present study showed that a considerable proportion of dairy and beef calves in this open water catchment region excreted Cryptosporidium (i.e. subgenotype IIaA18G3R1) and Giardia (e.g. assemblage A) that are consistent with those infecting humans, inferring that they are of zoonotic importance. Future work should focus on exploring, in a temporal and spatial way, whether these parasites occur in the environment and water of the catchment reservoir.
Water research, 2014
Protozoan pathogens present a significant human health concern, and prevention of contamination into potable networks remains a key focus for drinking water providers. Here, we monitored the change in Cryptosporidium concentration in source water during high flow events in a multi-use catchment. Furthermore, we investigated the diversity of Cryptosporidium species/genotypes present in the source water, and delivered an oocyst infectivity fraction. There was a positive and significant correlation between Cryptosporidium concentration and flow (ρ = 0.756) and turbidity (ρ = 0.631) for all rainfall-runoff events, despite variable source water pathogen concentrations. Cell culture assays measured oocyst infectivity and suggested an overall source water infectious fraction of 3.1%. No infectious Cryptosporidium parvum or Cryptosporidium hominis were detected, although molecular testing detected C. parvum in 7% of the samples analysed using PCR-based molecular techniques. Twelve Cryptospo...
Water quality and Cryptosporidium distribution in an upland water supply catchment, Cumbria, UK.
Four micro-catchment (MC) areas were identified to represent the main terrain types of a remote, sparsely populated upland valley catchment of 18 km2 in Cumbria, UK. These were improved land with good grazing (IB), steeply sloping land with rough grazing (SG), wet moorland with sparse grazing (WM) and enclosed woodland that excluded livestock and deer (EW). Each MC contained the origin of a small stream that flowed into Swindale Beck, the river draining the valley. The water quality during the 14-month study, as judged by chemical and physical parameters, was excellent, but it could not be regarded as pristine because of the frequent presence of Cryptosporidium oocysts arising from livestock and wild mammal faeces. Oocysts (0·2–5·6 l−1) detected by genus-specific immunofluorescent antibody were found in 32% of 188 water samples tested: ranking order EW 44%, IB 34%, Beck 30%, SG and WM 26%. Similarly, oocysts were identified in 9·5% of 1730 faecal samples. Small wild mammals (28%), calves (15·7%) and lambs (8·1%) were the dominant sources, whereas adult livestock (1·8%) and large wild mammals (4·8%) were less important. Autumn showed the highest occurrence of oocysts for both water and faecal samples. No hydrological controls were found to have a significant impact on the occurrence or concentration of oocysts in the main river or in the MCs, suggesting that their presence is controlled by seasonal changes in pathogen prevalence in the animal reservoir. Copyright © 2006 John Wiley & Sons, Ltd.
Journal of Water and Health, 2010
As part of investigations into the cause of a waterborne outbreak of Cryptosporidium hominis infection linked to a mains water supply, surface waters and wastewater treatment plants were tested for Cryptosporidium spp. Oocyst counts in base flow surface water samples ranged from nil to 29 per 10 l. Oocyst counts in effluent from a community wastewater treatment plant were up to 63 fold higher and breakout from one septic tank five logs higher. There were no peak (storm) flow events during the investigation. C. hominis, four named genotypes (cervine, muskrat II, rat, W19) and six new small subunit ribosomal RNA gene sequences were identified. Four of the new sequences were closely related to Cryptosporidium muskrat genotype I, one was closely related to the fox genotype and one to Cryptosporidium canis. C. hominis was found extensively in the catchment, but only at sites contaminated by wastewater, and in the treated water supply to the affected area. All were gp60 subtype IbA10G2, t...