Investigation of In Vitro Effectiveness of Polymeric Nanoparticles Containing Clinoptilolit on Cryptosporidium Parvum

Abstract

Cryptosporidium species, which do not have a definite prevention and treatment protocol, are pathogenic protozoans that cause diarrhoea in humans and many animal species. This study aimed to demonstrate the in vitro efficacy of clinoptilolite polymer nanoparticle, which has known antidiarrheal effects. DNA isolation was made for species identification of Cryptosporidium oocysts obtained from faeces samples from naturally infected calves and lambs. RFPL analysis was performed in typing. For this purpose, nested PCR and SrpI, VspI and MboII enzymes were used. The coverslip surface was covered with 80% CaCo-2 cells and infected with 1x109 C. parvum. Nanoparticles containing 250, 500, 750 and 1000 μg/mL clinoptilolite were applied to infected cells. Percent of infection rate was calculated by counting under a fluorescent microscope following incubation. While the infection rate was 23.46% in the water-treated control cell group, the percentage infection rates in the clinoptilolite-containing nanoparticle treated group were respectively 15.60%, 8.13%, 10.33% and 13.46%. Inhibition percentages were determined as 33.54%, 65.56%, 55.99% and 42.66%, respectively. As a result, it was observed that the nanoparticle containing clinoptilolite had anticriptocidal activity in infection with C. parvum in Caco-2 cells. In addition, it was observed that the efficacy was dose-dependent, and the IC50 value was the most appropriate value at 750 and 1000 μg/mL doses.