Research article

EFFECT OF GREEN ALGAE CHLORELLA EXTRACTION IN TREATMENT OF CRYPTOSPORIDIOSISI INFECTION IN EXPERIMENTAL INFECTED MICE

Russl Jalil Kazem El Heluo 1* May Naji Al Khanaq 2 and Wassan Hamza Mazal 3

Online First: December 18, 2022


The objective of this study was to conduct in vivo to detect the effect of alcoholic and aqueous Chlorella algea extracts in comparison with azithromycin on Cryptosporidium spp. and to examine its in male laboratory mice (Mus musculus) type Albino mice of c-Balb strain. Stool samples were collected from patients attending in Al-Kut Hospital suffering from diarrhea from December 2021 to end of March 2022, and 90 samples were examined for both sexes microscopic by using the modified Ziehl-Nelson Stain to detect infected the parasite's oocysts , Isolation and purification by flotation with Scheither's sugar solution and preserved in potassium dichromate for the purpose of using it in causing infection in laboratory mice. The experimental study was conducted on groups of 57 mice by oral administration of parasite oocysts at 104 oocyst / ml; while, the negative control group that drenched a physiological saline. The feces of mice inoculated with oocysts of parasites were microscopically examined using the modified Ziehl-Nelson Stain, as well as the molecular examination was conducted using Multiplex PCR technique. After the mice were divided into five groups with the uninfected and untreated group kept as a healthy negative control. The first group which included 21 mice was treated after it was divided into three subgroups A, B, C for each secondary group 7 mice they were treated with alcoholic extract of Chlorella at different concentrations 50, 100, and 150 mg/ml, respectively; while, the second group which included 21 mice on three groups A, B, and C was treated with aqueous extract of Chlorella at the previous concentrations for three consecutive days for each concentration. The third group was treated with azithromycin at a concentration of 500 ml, and the positive control group remained infected with the parasite and was not treated. After treatment a microscopic examination was performed by evaluating the excretion average of parasite oocysts using a hematocytometer slide. There was a decrease in the shedding of fecal oocysts in the groups treated with alcoholic extract at different concentrations, reaching down as 250 with the highest concentrations at the end of the treatment period. The average of shedding of fecal parasite oocysts of groups of mice at concentrations 50, 100, and 150 mg/ml reached 1.875, 625, and 250 oocyst / ml, respectively. The groups treated with the aqueous extract also showed a clear decrease in the shedding of fecal oocysts, as the average reached the three concentrations 50, 100, 150 mg/ml to 2.750, 1000, 700 oocyst/ml respectively, while the average of shedding of oocysts in the group treated with azithromycin reached 0 oocyst/ml in comparison with the infected positive control group in which the number of oocysts continued to increase to 14.863 oocyst/ml at the end of the treatment period, the results showed statistically significant differences between groups with different concentrations at P≤0.05. The results of the therapeutic efficacy of Chlorella alga extracts showed percentages of 87%, 95%, and 98% for the alcoholic extract at concentrations 50, 100, and 150 mg/ml, respectively; while the rates were recorded 81%, 93%, and 95% for aqueous extract at concentrations 50, 100, and 150 mg/ml. However, the therapeutic efficacy of azithromycin reached 100% at the end of the treatment period.

Keywords

Cryptosporidium spp., Chlorella Extract, Azithromycin