Microscopic, Molecular and Antigen Detection and Isolation of Cryptosporidium parvumParasites in Diarrheal Disease of Calves in Iran.

Abstract

Cryptosporidiosis is a parasitic disease caused by the protozoan Cryptosporidium in vertebrates. In livestock, especially ruminants, infants develop diarrheal syndromes. The infection is common worldwide , including Iran, where it is reported in several species. Morphological diagnosis of Cryptosporidium species is associated with many limitations and has no taxonomic value on its own, so the use of molecular methods can overcome these limitations to some extent. The present aims at microscopic, molecular and antigen detection and isolation of Cryptosporidium parvum parasites. Firstly, 300 samples were collected from different parts of Iran. Subsequently oocysts from feces were purified by the method of Casemore et al. using the flotation technique and stained by the modified Ziehl-Neelsen method (Henriksen method) and identified by diagnostic keys. ELISA test was also performed on the samples with results ranging from 1 to 4 positive. The results of our study show that, of the 300 cases tested for Cryptosporidium, 48 cases (16%) and 54 (18%) were positive in ELISA and PCR, respectively. Microscopic evaluation also mainly confirmed the ELISA results. These cases were collected in summer, autumn, and winter, with, more than 50% of the positive cases found among the samples collected in autumn. In addition, 54 positive cases were found by PCR test, which is 6 cases more than ELISA results. Finally, the results of PCR detection and ELISA were subjected to chi-square analysis, where no significant difference was found between the collected data (p=0.0587).