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Gut microbiome of Crocodylus porosus and cellular stress: inhibition of nitric oxide, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells

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Abstract

Crocodiles are renowned for their resilience and capacity to withstand environmental stressors, likely influenced by their unique gut microbiome. In this study, we determined whether selected gut bacteria of Crocodylus porosus exhibit anti-inflammatory effects in response to stress, by measuring nitric oxide release, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells. Using the Griess assay, the findings revealed that among several C. porosus gut bacterial isolates, the conditioned media containing the metabolites of two bacterial strains (CP27 and CP36) inhibited nitric oxide production significantly, in response to the positive control, i.e., taxol-treatment. Notably, CP27 and CP36 were more potent at reducing nitric oxide production than senloytic compounds (fisetin, quercetin). Using enzyme linked immunosorbent assays, the production of pro-inflammatory cytokines (IL-1β, TNF-α, PGE2), was markedly reduced by treatment with CP27 and CP36, in response to stress. Both CP27 and CP36 contain a plethora of metabolites to exact their effects [(3,4-dihydroxyphenylglycol, 5-methoxytryptophan, nifedipine, 4-chlorotestosterone-17-acetate, 3-phenoxypropionic acid, lactic acid, f-Honaucin A, l,l-Cyclo(leucylprolyl), 3-hydroxy-decanoic acid etc.], indicative of their potential in providing protection against cellular stress. Further high-throughput bioassay-guided testing of gut microbial metabolites from crocodiles, individually as well as in combination, together with the underlying molecular mechanisms, in vitro and in vivo will elucidate their value in the rational development of innovative therapies against cellular stress/gut dysbiosis.

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The data is available upon request to the corresponding author.

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Funding

This work was funded by the Air Force Office of Scientific Research (AFOSR), grant number: FA 8655-20-1-7004.

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Author notes

  1. Ruqaiyyah Siddiqui, Noor Akbar have contributed equally.

Authors and Affiliations

  1. College of Arts and Sciences, American University of Sharjah, 26666, Sharjah, United Arab Emirates

    Ruqaiyyah Siddiqui

  2. Microbiota Research Center, Istinye University, Istanbul, 34010, Turkey

    Ruqaiyyah Siddiqui & Naveed Ahmed Khan

  3. Research Institute of Medical and Health Sciences, University of Sharjah, 27272, Sharjah, United Arab Emirates

    Noor Akbar

  4. Centre for Discovery Brain Sciences, Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK

    Sutherland K. Maciver

  5. Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Taif University, 21944, Taif, Saudi Arabia

    Ahmad M. Alharbi

  6. Department of Medical Microbiology, Faculty of Medicine, Al-Baha University, 65799, Al-Baha, Saudi Arabia

    Hasan Alfahemi

Authors
  1. Ruqaiyyah Siddiqui
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  2. Noor Akbar
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  3. Sutherland K. Maciver
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  4. Ahmad M. Alharbi
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  5. Hasan Alfahemi
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  6. Naveed Ahmed Khan
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Contributions

RS and NAK conceptualized the study amid discussions with SKM. NA and RS carried out all experimental work amid critical discussions with NAK, AMA, HF, and SKM. RS prepared the first draft, while SKM, NA and NAK corrected and finalized the manuscript. All authors approved the final manuscript.

Corresponding author

Correspondence to Naveed Ahmed Khan.

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No animals or humans were used in this study.

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Communicated by Ran Wang.

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Siddiqui, R., Akbar, N., Maciver, S.K. et al. Gut microbiome of Crocodylus porosus and cellular stress: inhibition of nitric oxide, interleukin 1-beta, tumor necrosis factor-alpha, and prostaglandin E2 in cerebrovascular endothelial cells. Arch Microbiol 205, 344 (2023). https://doi.org/10.1007/s00203-023-03680-z

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  • DOI: https://doi.org/10.1007/s00203-023-03680-z

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