title: Prevalence and molecular characterization of Cryptosporidium spp. and Giardia duodenalis in diarrhoeic patients in the Qikiqtani Region, Nunavut, Canada.
abstract: BACKGROUND
  Although the prevalences of infection with the protozoan parasites Cryptosporidium spp. and Giardia duodenalis in humans appear to be relatively high in the Canadian North, their transmission patterns are poorly understood.
  
  
  OBJECTIVE
  To determine the detection rate and the molecular characteristics of Cryptosporidium spp. and Giardia duodenalis in diarrhoeic patients in the Qikiqtani (Baffin Island) Region of Nunavut, Canada, in order to better understand the burden of illness and the potential mechanisms of transmission.
  
  
  METHODS
  Diarrhoeal stool specimens (n=108) submitted to the Qikiqtani General Hospital for clinical testing were also tested for the presence of Cryptosporidium spp. and Giardia duodenalis using epifluorescence microscopy and polymerase chain reaction (PCR). DNA sequencing and restriction fragment length polymorphism (RFLP) analyses were performed on PCR-positive specimens to determine the species, genotypes and sub-genotypes of the parasites.
  
  
  RESULTS
  Cryptosporidium was detected in 15.7% of the diarrhoeic patients, while Giardia was detected in 4.6%. DNA sequencing of a fragment of the small subunit rRNA gene indicated that all of the Cryptosporidium amplicons had a 100% homology to C. parvum, and a gp60 assay showed that all aligned with C. parvum sub-genotype IIa. Microsatellite analysis revealed 3 cases of sub-genotype IIaA15G2R1, 2 of IIaA15G1R and 1 case each of sub-genotypes IIaA16G1R1 and IIaA15R1. For Giardia, results based on the amplification of both the 16S rRNA gene and the gdh gene were generally in agreement, and both DNA sequencing and RFLP demonstrated the presence of the G. duodenalis Assemblage B genotype.
  
  
  CONCLUSIONS
  Both C. parvum and G. duodenalis Assemblage B were present in human diarrhoeal stool specimens from Nunavut, which was suggestive of zoonotic transmission, although human-to-human transmission cannot be ruled out. To fully understand the public health significance of the different Cryptosporidium and Giardia species and genotypes in diarrhoeic patients, it will be imperative to establish the extent of genetic diversity within these parasites through comprehensive studies of the molecular epidemiology of cryptosporidiosis and giardiasis in the Nunavut region.
authors.affiliation: Bureau of Microbial Hazards, Food Directorate, Health Canada, Ottawa, ON, Canada.
authors.forename: Asma
authors.initials: A
authors.lastname: Iqbal
authors.affiliation: Department of Pathology, British Columbia Children's Hospital, Vancouver, BC, Canada.
authors.forename: David M
authors.initials: DM
authors.lastname: Goldfarb
authors.affiliation: Department of Pathology and Laboratory Medicine, Children's Hospital of Eastern Ontario, University of Ottawa, Ottawa, ON, Canada.
authors.forename: Robert
authors.initials: R
authors.lastname: Slinger
authors.affiliation: Bureau of Microbial Hazards, Food Directorate, Health Canada, Ottawa, ON, Canada; brent.dixon@hc-sc.gc.ca.
authors.forename: Brent R
authors.initials: BR
authors.lastname: Dixon
mesh_terms: D000818:Animals; D002170:Canada / Q000453:epidemiology; D003081:Cold Climate; D003457:Cryptosporidiosis / Q000175:diagnosis / Q000453:epidemiology; D003458:Cryptosporidium / Q000302:isolation & purification*; D003967:Diarrhea / Q000453:epidemiology* / Q000469:parasitology*; D005243:Feces / Q000469:parasitology*; D005260:Female; D016829:Giardia lamblia / Q000302:isolation & purification*; D005873:Giardiasis / Q000175:diagnosis / Q000453:epidemiology; D006801:Humans; D008297:Male; D020957:Nunavut / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence; D012494:Sampling Studies
keywords: Canadian North; Cryptosporidium parvum; Giardia duodenalis; Nunavut; PCR; diarrhoea; genotyping; microscopy; parasites