Investigation of in vitro effectiveness of polymeric nanoparticles containing clinoptilolit on cryptosporidium parvum

Cryptosporidium spp is an enteric protozoan pathogen causing diarrhea in human and multi-species species. Several drugs have been used to treat Cryptosporidium parvum infection; However, a complete parasitological recovery has not been achieved. Currently, there is no definitive protection and treatment protocol for C. parvum. In this context, a new treatment protocol for cryptosporidiosis is needed. Antiviral, antitumoral and antidiarrheal activities of clinoptilolite investigated and found to have positive effects on these fields. In this study, it was aimed to reveal the in vitro efficacy of polymer nanoparticle containing clinoptilolite on C. parvum. In this study, human colon adenocarcinoma (Caco-2) cells were used to perform experimental in vitro infection. DNA isolation was performed to determine the strain and type of Cryptosporidium oocysts from natural infected calves or lambs. Typing of Cryptosporidium parvum was performed by RFLP analysis using nested PCR and SspI, VspI and MboII enzymes. Human colon adenocarcinoma (Caco-2) cells were used to perform experimental in vitro infection. In infectious experiments, 18 mm sterile round laminates were placed into 12 well cell culture plates and Caco-2 cells were plated on them. Cells were coated with 80% of the surface of the coverslips and then infected with 1x106 C. parvum oocysts. Clinoptilolite-containing polymer nanoparticles and water in a group were added to the infected enterocytes at 4 different concentrations (250, 500, 750, 1000 μg / ml) and the control group was formed. Percent infection rate was determined by counting oocysts in cells and cells in 20 side-by-side microscope on the same line at 400x magnification on a fluorescence microscope after incubation. Statistical evaluation of the data was performed using the Statistical Package for the Social Sciences (SPSS) 19 package program. p <0.05 was considered significant in all statistical evaluations. In this context, the percentage of infection occurrence in the control group was determined as 23,46%. Different concentrations of clinoptilolite-containing polymer nanoparticles (1000μg / ml, 750μg / ml, 500μg / ml, 250μg / ml) was determined as 15.6%, 8.13%, 10.33% and 13.46% respectively. It was revealed that the percent inhibition was 33.54%, 65.56%, 55.99%, 42.66% of the inhibition percentage at different concentrations of the clinopitilolite nanoparticle (1000μg / ml, 750μg / ml, 500μg / ml, 250μg / ml). In conclusion, anticryptocidal activity of in vitro infection of C. parvum with Caco-2 cells, clinoptilolite-containing polymer nanoparticles was detected. However, it was determined that this activity was dose-related and the concentrations of clinoptilolite-containing polymer nanoparticles with inhibition percentage (IC50) values were 750μg / ml and 500μg / ml. Key words: Cryptosporidium parvum, clinoptilolite, nanoparticle.