Anti- Cryptosporidium parvum activity of Artemisia judaica L. and its fractions: in vitro and in vivo assays

Shahira A Ahmed; Enas E Eltamany; Mohamed S Nafie; Sameh S Elhady; Panagiotis Karanis; Amira B Mokhtar

doi:10.3389/fmicb.2023.1193810

Anti- Cryptosporidium parvum activity of Artemisia judaica L. and its fractions: in vitro and in vivo assays

Front Microbiol. 2023 Jul 3:14:1193810. doi: 10.3389/fmicb.2023.1193810. eCollection 2023.

Authors

Shahira A Ahmed¹, Enas E Eltamany², Mohamed S Nafie³, Sameh S Elhady^{4

5}, Panagiotis Karanis^{6

7}, Amira B Mokhtar¹

Affiliations

¹ Department of Medical Parasitology, Faculty of Medicine, Suez Canal University, Ismailia, Egypt.
² Department of Pharmacognosy, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
³ Department of Chemistry (Biochemistry Program), Faculty of Science, Suez Canal University, Ismailia, Egypt.
⁴ Department of Natural Products, Faculty of Pharmacy, King Abdulaziz University, Jeddah, Saudi Arabia.
⁵ Center for Artificial Intelligence in Precision Medicines, King Abdulaziz University, Jeddah, Saudi Arabia.
⁶ University of Cologne, Medical Faculty and University Hospital, Cologne, Germany.
⁷ Department of Basic and Clinical SciencesUniversity of Nicosia Medical School, Nicosia, Cyprus.

Abstract

Background: This study investigates the toxic activity of Artemisia judaica ethanolic extract (ArEx) as well as its phenolic fraction (ArPh), and terpenoid fraction (ArT) against Cryptosporidium parvum (C. parvum) oocysts.

Methods: Over a 4 months period, estimation of the total phenolic (TPC), total flavonoids (TFC), and total terpenoids contents (TTC) in ArEx; investigation of the in vitro antioxidant activity of ArEx, ArPh, and ArT; evaluation of ArEx, ArPh, and ArT toxic activity against C. parvum oocysts using MTT assay; parasitological analysis on ArPh-treated C. parvum oocysts and comet assay were performed both in vitro and in vivo (infectivity).

Results: The ArEx TPC, TFC, and TTC was 52.6 ± 3.1 mgGAE/g, 64.5 ± 3.1 mg QE/g, and 9.5 ± 1.1 mg Linol/g, respectively. Regarding the phytochemical in vitro antioxidant activity, the ArPh exhibited the highest antioxidant activity compared to the ArEx and ArT. The ArPh showed promising free radical scavenging activity of DPPH and ABTS^•+ with IC₅₀ values of 47.27 ± 1.86 μg/mL and 66.89 ± 1.94 μg/mL, respectively. Moreover, the FRAP of ArPh was 2.97 ± 0.65 mMol Fe⁺²/g while its TAC was 46.23 ± 3.15 mg GAE/g. The ArPh demonstrated toxic activity against C. parvum oocysts with a potent IC₅₀ value of 31.6 μg/mL compared to ArT (promising) and ArEx (non-effective). ArPh parasitological analysis demonstrated MIC₉₀ at 1000 μg/ml and effective oocysts destruction on count and morphology. ArPh fragmented oocysts nuclear DNA in comet assay. Beginning at 200 μg/mL, ArPh-treated oocysts did not infect mice.

Conclusion: To combat C. parvum infection, the phenolic fraction of A. judaica L. shows promise as an adjuvant therapy or as a source of potentially useful lead structures for drug discovery.

Keywords: Artemisia judaica L.; Cryptosporidium sp.; MTT assay; comet assay; drug discovery and public health; in vitro; in vivo.