Abstract

• Verbena officinalis is an intrinsic supplier for biologically active metabolites comprising flavonoids, phenolic acids, organic acids, phenylethanoids, and coumarins. • Verbena officinalis n -butanol extract was proved to have an effective anti-cryptosporidial activity in the infected immunocompromised mice exceeding the gold standard drug nitazoxanide. • Verbena officinalis n -butanol extract was reported to have a promising anti-inflammatory effect, alleviating pathological changes in the small intestine and the liver of infected immunocompromised mice, exceeding the gold standard drug nitazoxanide. • The combination of Verbena officinalis n -butanol extract, and nitazoxanide gave the best results. Cryptosporidiosis is a global zoonotic infection that causes water-borne epidemics of diarrhea. Nevertheless, there are few available therapies for cryptosporidiosis. However, the gold standard drug nitazoxanide (NTZ) has limited efficacy in malnourished and immunocompromised patients. Furthermore, Verbena officinalis L. is a herbal plant widely used in traditional medicine to cure several health disorders and is recognized to possess numerous therapeutic applications. In the present study, the phytochemical composition of aerial part extract from Verbena officinalis was investigated via LC-ESI-MS/MS. Furthermore, the anti-cryptosporidial activity was also performed using an animal model. Fifty mice were divided into 5 groups; GI: non-infected (Negative control), GII: infected non treated (positive control), GIII: infected, treated with NTZ, GIV: infected, treated with V. officinalis n -butanol extract , GV: infected, treated with a combination of NTZ and V. officinalis. Parasitological examination revealed a highly significant difference ( P -value < 0.001) between GIII, GIV, and GV compared to GII regarding the mean number of Cryptosporidium spp. oocyst in the stool. Moreover, GV showed the best efficacy with a percentage of 87%. Also, histopathological examination showed variable degrees of improvement in the villous broadening, and the inflammatory infiltrates in the small intestine with a reduction of hepatocyte degeneration and mononuclear infiltration in GIII, GIV, and GV compared to GII, with the best results seen in GV. Additionally, the chemical profiling of n -butanol extract identified 16 secondary metabolites comprising flavonoids, phenolic acids, phenylethanoids, and coumarins. In conclusion, V. officinalis is an intrinsic supplier of biologically active metabolites with outstanding anti-parasitic and possible anti-inflammatory effects.

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