Differential Effect of Paromomycin and Nitazoxanide on Clinical Isolates of Cryptosporidia In Vitro.

ABSTRACT

Background: Cryptosporidiosis is one of the most difficult protozoan infections to treat, with only two drugs i.e. nitazoxanide and paromomycin known for treatment with variable response in different patients. Human cryptosporidiosis is accounted mainly by C. hominis and C. parvum. These two species or their subtypes are known to differ in clinical manifestations, and may differ in their response to drugs. So, we planned the study to see the effect of nitazoxanide and paromomycin on different isolates of Cryptosporidium in vitro. Methods: MDCK cell lines were used for in vitro growth of parasite and cytotoxicity of drugs to MDCK cells was determined by MTT assay after 3, 12 and 24 hours of drug exposure. Efficacy of non-toxic drug concentrations (<25% cytotoxic) on 12 Cryptosporidium isolates (7 C. hominis and 5 C. parvum) was determined at three different life cycle stages (in vitro growth, invasion and oocyst) by quantitative RT-PCR. Unpaired t-test was used to calculate the difference response of Cryptosporidium isolates to nitazoxanide and paromomycin. Results and conclusions: Cytotoxicity of nitazoxanide and paromomycin increased in dose and time dependent manner. After 24 hours of drug exposure, >25% cytotoxic effect was seen with nitazoxanide and paromomycin at concentrations of more than, 25μg/ml and 6mg/ml, respectively. Nitazoxanide was more effective than paromomycin in decreasing in vitro growth, invasion inhibition and reducing oocyst viability of Cryptosporidium isolates. Drugs effect was higher on growth inhibition followed by invasion inhibition and least in decreasing oocyst viability. Different isolates had variable response to drugs; cumulatively C. parvum isolates were more susceptible at particular drug concentrations than C. hominis isolates.