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Peer-reviewedCC BY 4.0PLOS Neglected Tropical Diseases

Unravelling the synergistic potential of Olibanum extract and Bacillus clausii in combating intestinal cryptosporidiosis in immunosuppressed mice

  1. 1Department of Medical Parasitology, Faculty of Medicine, Alexandria University, Alexandria, Egypt
  2. 2Department of Chemistry, Faculty of Science, Alexandria University, Alexandria, Egypt
  3. 3Department of Pathology, Faculty of Medicine, Alexandria University, Alexandria, Egypt

Abstract

Given the grave risk posed by intestinal cryptosporidiosis to vulnerable populations, the current research focused on a new therapeutic avenue. Olibanum (OL) extract and probiotic Bacillus clausii (B. clausii) and their combination were evaluated against Cryptosporidium in vivo in immunosuppressed mice via parasitological, ultrastructural, biochemical, and histopathological analysis. Liquid chromatography-mass spectrometry (LC–MS) analysis of OL revealed the predominance of incensole isomers in its chemical composition. Regarding oocyst count in stool, the highest percentage of oocyst reduction was achieved after combined therapy (98.4%) on the 19th day post-infection, followed by B. clausii-treated group (87.6%), then the OL-treated group (87%). Similar ultrastructural alterations were noticed by scanning electron microscopy (SEM) following treatment by both OL and B. clausii, ranging from shrunken oocysts to severely deformed ones with extensive pits or protrusions. Nonetheless, their combination resulted in complete distortion of the oocysts, leading to their rupture. Biochemically, both OL and B. clausii treatment, either alone or in combination, provoked a decrease in the serum MDA levels, together with a corresponding rise in the mean levels of GSH. Concerning the histopathological findings, the best results were achieved following the combined OL and B. clausii treatment, which has restored villus architecture and remarkably ameliorated detrimental intestinal inflammation and goblet cell count. Finally, immunohistochemical analysis of the lymphocytic population of the intestinal villi revealed a statistically significant rise in the CD4+ /CD8+ ratio after treatment with either OL, B. clausii or their combination, which signals their proficient use for this therapeutic endeavour. Author summary: Cryptosporidiosis is a cosmopolitan food, and waterborne infection caused mainly by the obligate apicomplexan parasite, Cryptosporidium parvum (C. parvum). Cryptosporidiosis is responsible for debilitating diarrhea outbreaks, especially in children. Additionally, it is a major opportunistic infection that may lead to devastating chronic disease specially among immunocompromised patients. Current therapy of cryptosporidiosis depends on nitazoxanide (NTZ), the only FDA-approved medication for treatment of cryptosporidiosis. However, treatment failure of NTZ, especially in immunocompromised individuals, has been widely reported. Therefore, there is an urgent need to develop high efficiency and low-toxicity therapies against Cryptosporidium. Recently, phytotherapy and probiotics have unprecedentedly revolutionized the medical research, owing to their well-characterized pharmacological and safety profiles. Conspicuously, the present study presents significant findings on the promising therapeutic efficacy of Olibanum (OL) extract and probiotic Bacillus clausii (B. clausii) and their combinationin on cryptosporidiosis in immunosuppressed murine model. This efficacy was supported by significant reduction in the parasite burden, improved intestinal histopathological changes, obvious ultrastructural alternations in oocysts with significant antioxidant, immunomodulatory activities.

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BibTeX
@article{arafa2026unravelling,
  title = {{Unravelling the synergistic potential of Olibanum extract and Bacillus clausii in combating intestinal cryptosporidiosis in immunosuppressed mice}},
  author = {Fadwa M. Arafa and Mohamed Hagar and Ahmed Zakaria and Eman Sheta and Nehal N. Hezema},
  journal = {PLOS Neglected Tropical Diseases},
  year = {2026},
  month = sep,
  doi = {10.1371/journal.pntd.0014734},
  url = {https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0014734},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - Unravelling the synergistic potential of Olibanum extract and Bacillus clausii in combating intestinal cryptosporidiosis in immunosuppressed mice
AU  - Fadwa M. Arafa
AU  - Mohamed Hagar
AU  - Ahmed Zakaria
AU  - Eman Sheta
AU  - Nehal N. Hezema
PY  - 2026
DA  - 2026/09/30
JO  - PLOS Neglected Tropical Diseases
DO  - 10.1371/journal.pntd.0014734
UR  - https://journals.plos.org/plosntds/article?id=10.1371/journal.pntd.0014734
AB  - Given the grave risk posed by intestinal cryptosporidiosis to vulnerable populations, the current research focused on a new therapeutic avenue. Olibanum (OL) extract and probiotic Bacillus clausii (B. clausii) and their combination were evaluated against Cryptosporidium in vivo in immunosuppressed mice via parasitological, ultrastructural, biochemical, and histopathological analysis. Liquid chromatography-mass spectrometry (LC–MS) analysis of OL revealed the predominance of incensole isomers in its chemical composition. Regarding oocyst count in stool, the highest percentage of oocyst reduction was achieved after combined therapy (98.4%) on the 19th day post-infection, followed by B. clausii-treated group (87.6%), then the OL-treated group (87%). Similar ultrastructural alterations were noticed by scanning electron microscopy (SEM) following treatment by both OL and B. clausii, ranging from shrunken oocysts to severely deformed ones with extensive pits or protrusions. Nonetheless, their combination resulted in complete distortion of the oocysts, leading to their rupture. Biochemically, both OL and B. clausii treatment, either alone or in combination, provoked a decrease in the serum MDA levels, together with a corresponding rise in the mean levels of GSH. Concerning the histopathological findings, the best results were achieved following the combined OL and B. clausii treatment, which has restored villus architecture and remarkably ameliorated detrimental intestinal inflammation and goblet cell count. Finally, immunohistochemical analysis of the lymphocytic population of the intestinal villi revealed a statistically significant rise in the CD4+ /CD8+ ratio after treatment with either OL, B. clausii or their combination, which signals their proficient use for this therapeutic endeavour. Author summary: Cryptosporidiosis is a cosmopolitan food, and waterborne infection caused mainly by the obligate apicomplexan parasite, Cryptosporidium parvum (C. parvum). Cryptosporidiosis is responsible for debilitating diarrhea outbreaks, especially in children. Additionally, it is a major opportunistic infection that may lead to devastating chronic disease specially among immunocompromised patients. Current therapy of cryptosporidiosis depends on nitazoxanide (NTZ), the only FDA-approved medication for treatment of cryptosporidiosis. However, treatment failure of NTZ, especially in immunocompromised individuals, has been widely reported. Therefore, there is an urgent need to develop high efficiency and low-toxicity therapies against Cryptosporidium. Recently, phytotherapy and probiotics have unprecedentedly revolutionized the medical research, owing to their well-characterized pharmacological and safety profiles. Conspicuously, the present study presents significant findings on the promising therapeutic efficacy of Olibanum (OL) extract and probiotic Bacillus clausii (B. clausii) and their combinationin on cryptosporidiosis in immunosuppressed murine model. This efficacy was supported by significant reduction in the parasite burden, improved intestinal histopathological changes, obvious ultrastructural alternations in oocysts with significant antioxidant, immunomodulatory activities.
SN  - 1935-2735
ER  -