Browsing by Subject "Cryptosporidium parvum"
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(2018)Cryptosporidium parvum is a coccidian parasite. It causes an intestinal infection, named cryptosporidiosis. C. parvum infects mainly young calves and can be transmitted zoonotically to humans. C. parvum causes watery diarrhea, which is usually self-limiting. Cryptosporidiosis can be life-threatening to small children, immunocompromised patients and weak calves. C. parvum spreads mainly by the fecal-oral route by oocysts. It multiplies intracellularly, usually in the enterocytes of the small intestine. The life cycle is spent in a single host. Oocysts are extremely resilient and can cause large epidemics when household water or food gets exposed. Aim of this study was to research subtypes of protozoan C. parvum that occurred in calves in Finland 2012-2017, and research their divergence geographically and temporally. Additionally subtypes of 10 C. parvum -isolates from humans were studied and results were compared to subtypes found in calves. Fassisi BoDia, a rapid test designed to identify the four most common pathogens that infect young calves (rotavirus, coronavirus, C. parvum and E. coli K99/F5-type) was tested. The rapid test also detects possible mixed infections. Test is immunocromatographic and the diagnostic method to find C. parvum’s specific protein is based on monoclonal antibodies. Sample collection for determination of subtypes consists of 100 fecal samples from calves and 10 human fecal samples. Fassisi BoDia rapid test was tested with 67 fecal samples from calves. Subtyping was made by multiplying the protein gp60 coding gene with nested-PCR and by analyzing sequences of the gene. Research was made in Evira research unit of veterinary bacteriology and pathology in Kuopio. Subtype IIaA15G2R1 was the most common in calves. Distribution of this subtype had increased in Finland after subtype results from earlier years. Subtype IIaA15G2R1 was mainly found in western Finland, especially around Pohjanmaa. Subtype IIaA18G1R1 was the only one found in human samples, the same subtype was not found from calf samples in this study. Sensitivity of the Fassisi BoDia rapid test for C. parvum was 91,7 % and specificity was 83,9 % reliability.
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(2021)Cryptosporidium, a parasitic eukaryote, is the causative agent of cryptosporidiosis, a diarrheal disease with the potential to be life threatening in immunocompromised people. The number of cases has been rising as diagnostics have improved, suggesting the disease is more common than previously thought. Currently in Finland, samples are only typed to the genus level. The objective of the study was to establish a Cryptosporidium typing protocol for outbreak investigation nationally at the Finnish Institute of Health and Welfare with the long-term goal of preventing outbreaks. Reliable typing methods could also help find new reasons behind the increase in cases, monitor the emergence of new subtypes, and reveal new transmission routes. Typing was set up with samples from the Helsinki and Uusimaa Hospital District laboratory as well as the Finnish Food Authority. DNA was extracted from stool samples, species were identified with real-time PCR of the 18S rRNA gene, and gp60 subtypes were determined with nested PCR as well as Sanger sequencing. The Zymo QuickDNA Fecal/Soil Microbe Miniprep Kit was suitable for extracting troublesome parasite DNA. Out of the successfully subtyped samples, the majority was identified as Cryptosporidium parvum, out of which 60 % belonged to the IIaA15G2R1 subtype. The typing method described in this study was successful in typing 19 of 22 samples and will be utilised by the Finnish Institute of Health and Welfare for outbreak investigation on the national level.
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