Differential Effect of Paromomycin and Nitazoxanide on Clinical Isolates of Cryptosporidia In Vitro.
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Date
2014-10
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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.
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Keywords
MDCK, q-RT-PCR, oocyst viability, invasion-inhibition, C. hominis, subgenotype
Citation
Sharma Poonam, Sharma Aman, Sehgal Rakesh, Khurana Sumeeta. Differential Effect of Paromomycin and Nitazoxanide on Clinical Isolates of Cryptosporidia In Vitro. International Journal of Applied Biology and Pharmaceutical Technology. 2014 Oct-Dec; 5(4): 134-146.