Biosynthesized Zinc Oxide Nanoparticles Regulating Immune Response and Liver Enzymes in Reducing Inflammation from Cryptosporidiosis-induced Infection

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Abstract
Background: Cryptosporidium, a global intestinal protozoan parasite, induces mild to severe diarrhea in several vertebrate hosts, potentially resulting in life-threatening sickness. This study aimed to assess the efficacy of biosynthesized nano-zinc oxide using cellulose nanocrystals and Zingiber officinale extract (Bio-ZnNPs) in the treatment of experimentally infected mice with cryptosporidiosis. Methods: A total of 20 male mice were allocated into four experimental groups (G1-G4), with triplicates. G1 (Control); non-infected, non-treated, G2 (Inf); infected with Cryptosporidium parvum, G3 (Bio-ZnNPs); infected- treated with Bio-ZnNPs at 200 mg/kg B.W. and G4 (NTZ) infected- treated with nitazoxanide (NTZ) at 100 mg/kg B.W. Mice were inoculated with 3000 oocysts of C. parvum. All treatments commenced on the initial day of oocyst shedding and persisted for five continuous days for all groups, excluding the negative control one. Oocysts count was assessed daily from the 1st to the 5th day post-treatment. Hematological analyses, blood biochemistry, immunoglobulin level assessments and histopathological evaluations were performed on day 5 post-treatment. Result: The Bio-ZnNPs treated group exhibited the highest percentage reduction in C. parvum oocyst excretion. Furthermore, Bio-ZnNPs treatment resulted in notable enhancements in hematological parameters, liver enzymes and immunoglobulin levels, including immunoglobulin G and immunoglobulin M using Bio-ZnNPs. Additionally, a significant improvement in intestinal lesions with no intraluminal oocysts was noted in the Bio-ZnNPs treated group. Taken all together, this work demonstrated the promising performance of nanocomposites composed of cellulose nanocrystals and zinc oxide nanoparticles as an effective drug delivery strategy to enhance the efficacy of Zingiber officinale extract against experimental infected mice with C. parvum.

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