Therapeutic Potency of Ginger, Garlic, and Pomegranate Extracts Against Cryptosporidium parvum-Mediated Gastro-Splenic Damage in Mice

Abstract

Purpose: Cryptosporidium parvum is a protozoan parasite infecting most mammalian hosts and causing major health issues. The present study investigated the efficacy of ginger (Zingiber officinale), garlic (Allium sativum), and pomegranate (Punica granatum) peel extracts on the development and progression of experimental cryptosporidiosis in mice.

Methods: Eighty-two mice were assigned to 6 groups: control, infected non-treated, metronidazole (MTZ), ginger, garlic, and pomegranate. The control group topically received no treatments. The infected non-treated group was experimentally infected by 10⁴ C. parvum oocysts per mouse using a stomach tube. The MTZ group was infected with C. parvum oocysts combined with MTZ (50 mg/kg b.w./day). The ginger, garlic, and pomegranate groups daily received different plant extracts at doses of 100 mg/kg BW, 50 mg/kg BW, and 3 gm/kg BW, respectively, followed by infection with C. parvum oocysts. All treatments were applied orally one day after the infection for continuous 30 days.

Results: Histopathological and immunohistochemical examinations for P53 and caspase-3 expressions in stomach and spleen tissues showed that MTZ and garlic-treated mice had a more significant effect on infected mice.

Conclusion: The garlic extract was found to exert a more pronounced effect on infected mice compared with the other treatments as well as to improve health. Garlic extracts, therefore, represent an effective and natural therapeutic alternative for the treatment of cryptosporidiosis with low side effects and without drug resistance.

Keywords: Caspase-3; Cryptosporidium parvum; Garlic; Ginger; P53; Pomegranate.

Fig. 1

Histopathological sections of gastric mucosa showed: (A) Normal control with normal gastric glands. (B1, B2) Infected control showed marked desquamation of the lining epithelium of the gastric glands associated with the presence of oocysts (arrows head). (C) Infected animal treated with MTZ showed mild degenerative lesions of the gastric lining epithelial cells and few oocysts. (E) Infected animal treated with ginger showed degenerative changes within the parietal cells with the presence of oocysts within the epithelial cell, (F) Infected animal treated with garlic showed mild degenerative lesions of the gastric lining epithelial cells and few oocysts. (G) Infected animal treated with pomegranate showed decrease in the epithelial lining degeneration and the presence of oocysts within gastric glands. (1H) Lesion scoring of gastric mucosa in different groups. Data presented as Mean ± SE (n = 5), analyzed using one way ANOVA at p ≤ 0.05, *significance compared to control

Fig. 2

Histopathological sections of spleen showing A Spleen of control animal showed normal splenic architecture with normal and white pulps. B Spleen of infected untreated mice showed amyloidosis (arrows) associated with severe lymphoid depletion. C Spleen of MTZ-treated group showed lymphoid hyperplasia, D spleen of ginger-treated mice showed mild to moderate degree of lymphoid depletion, E spleen of garlic-treated group showed lymphoid hyperplasia associated with macrophages activation within the follicle, F spleen of pomegranate-treated group showed mild lymphoid depletion within the follicle associated with macrophages activation. 2H Lesion scoring of different groups. Data presented as Mean ± SE (n = 5), analyzed using one way ANOVA at p ≤ 0.05, *significance compared to control

Fig. 3

Immunohistochemical sections investigation revealed negative expression of P53 in glandular gastric mucosa of the control group (A). Significant positive brown nuclear expression was demonstrated in glandular cells (black arrows) in an infected group (B). Markedly decreased positive brown nuclear expression in glandular cells (black arrows) in ginger and pomegranate-treated groups (D, F), and become weak in glandular cells (black arrows) in MTZ and garlic-treated groups (C, E). 3H Quantification of P53 in the gastric mucosa in different groups. Data presented as Mean ± SE (n = 5), analyzed using one way ANOVA at p ≤ 0.05, *significance compared to control (colour figure online)

Fig. 4

Immunohistochemical sections investigation revealed mild positive brown cytoplasmic expression of caspase-3 was detected in glandular gastric mucosa of the normal control group (A). Significantly increased positive brown cytoplasmic expression appears in glandular cells (black arrows) in an infected group (B). Markedly decreased positive brown cytoplasmic expression in glandular cells (black arrows) ginger and pomegranate treated groups (D, F), and become weak in glandular cells (black arrows) in MTZ and garlic treated groups (C, E). 4H Quantification of caspase-3 in the gastric mucosa in different groups. Data presented as Mean ± SE (n = 5), analyzed using one way ANOVA at p ≤ 0.05, *significance compared to control (colour figure online)