Cryptosporidium immunolabeling in paraffinembedded ileum of CD-1 neonatal mice


Mohamed Mammeri, Hélène Huet, Christine Julien, Isabelle Vallée, Nathalie Cordonnier, Karim Tarik Adjou

Cryptosporidiosis is a well-recognized zoonotic disease and represents both a major public health and veterinary problem. There are currently no available commercial antibodies designed for Cryptosporidium labeling in histological sections, and no protocol has been suggested. The aim of this study was to establish a standardized protocol using immunofluorescence techniques to detect Cryptosporidium in paraffin-embedded intestine sections. Neonatal CD-1 mice were experimentally infected with 105 Cryptosporidium parvum Iowa strain oocysts. At peak infection (8 days postinfection), intestines were removed, fixed in formalin buffer and included in paraffin. Three commercial anti-Cryptosporidium antibodies (Sporo-Glo™ , Crypt-a-Glo™ , and Merifluor® ) were tested on tissue sections using standard immunofluorescence methods. Both parasite colonization and nonspecific immunolabeling in the terminal ileum were evaluated. Our results showed that all antibodies detected intracellular developmental stage C. parvum. Qualitative differences were noted between antibodies, particularly in relation to background staining and artefacts, which were more present with Crypt-a-Glo™ and the Merifluor® antibodies. Our results indicated that Sporo-Glo™ was the most effective antibody for Cryptosporidium detection in paraffin-embedded intestine. This technique proved to be an efficient tool for histological studies of cryptosporidiosis and could be recommended in future studies investigating infectivity of Cryptosporidium isolates, as well as testing efficacy of new therapeutic agents

Share this article