Journal Article

Molecular characterization of Cryptosporidium isolates from humans in Ontario, Canada

Lau, Rachel, Guy, Rebecca A., Boggild, Andrea K., Thivierge, Karine, Yanta, Christine A., Rankin, Marisa A., Muchaal, Pia K.

2021

Overview

Background Cryptosporidiosis is a gastrointestinal disease with global distribution. It has been a reportable disease in Canada since 2000; however, routine molecular surveillance is not conducted. Therefore, sources of contamination are unknown. The aim of this project was to identify species and subtypes of Cryptosporidium in clinical cases from Ontario, the largest province in Canada, representing one third of the Canadian population, in order to understand transmission patterns. Methods A total of 169 frozen, banked, unpreserved stool specimens that were microscopy positive for Cryptosporidium over the period 2008–2017 were characterized using molecular tools. A subset of the 169 specimens were replicate samples from individual cases. DNA was extracted directly from the stool and nested PCR followed by Sanger sequencing was conducted targeting the small subunit ribosomal RNA (SSU) and glycoprotein 60 ( gp60 ) genes. Results Molecular typing data and limited demographic data were obtained for 129 cases of cryptosporidiosis. Of these cases, 91 (70.5 %) were due to Cryptosporidium parvum and 24 (18.6%) were due to Cryptosporidium hominis . Mixed infections of C. parvum and C. hominis occurred in four (3.1%) cases. Five other species observed were Cryptosporidium ubiquitum ( n = 5), Cryptosporidium felis ( n = 2), Cryptosporidium meleagridis ( n = 1), Cryptosporidium cuniculus ( n = 1) and Cryptosporidium muris ( n = 1). Subtyping the gp60 gene revealed 5 allelic families and 17 subtypes of C. hominis and 3 allelic families and 17 subtypes of C. parvum . The most frequent subtype of C. hominis was IbA10G2 (22.3%) and of C. parvum was IIaA15G2R1 (62.4%). Conclusions The majority of isolates in this study were C. parvum , supporting the notion that zoonotic transmission is the main route of cryptosporidiosis transmission in Ontario. Nonetheless, the observation of C. hominis in about a quarter of cases suggests that anthroponotic transmission is also an important contributor to cryptosporidiosis pathogenesis in Ontario. Graphical Abstract

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adolescent / Adult / Aged / Aged, 80 and over / Biomedical and Life Sciences / Biomedicine / Child / Child, Preschool / Coccidiosis / Contamination / Cryptosporidiosis / Cryptosporidiosis - epidemiology / Cryptosporidiosis - parasitology / Cryptosporidiosis - transmission / Cryptosporidium / Cryptosporidium - classification / Cryptosporidium - genetics / Cryptosporidium - isolation & purification / Cryptosporidium hominis / Cryptosporidium meleagridis / Cryptosporidium muris / Cryptosporidium parvum / Deoxyribonucleic acid / Developing countries / Disease transmission / DNA / DNA, Protozoan - genetics / Entomology / Ethics / Feces - parasitology / Female / Food contamination & poisoning / Gastrointestinal diseases / Gene loci / genes / Genetic aspects / Genetic Variation / genomics and proteomics / Genotype / geographical distribution / Glycoproteins / Gp60 / Health aspects / Humans / Identification and classification / Infections / Infectious Diseases / Laboratories / LDCs / Male / Medical research / Medicine, Experimental / Microscopy / Middle Aged / monitoring / notifiable disease / Nucleic acids / Nucleotide sequence / Ontario / Ontario - epidemiology / Parasite genetics / Parasites / Parasitology / Pathogenesis / PCR / Phylogeny / Polymerase chain reaction / Public health / ribosomal RNA / RNA / rRNA / Sanger sequencing / Sequence Analysis, DNA / Tropical Medicine / Veterinary Medicine/Veterinary Science / Virology / Young Adult / Zoonoses / Zoonosis