Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry

Research output: Contribution to journalArticleResearchpeer review

Authors

  • Usman Majeed
  • Afshan Shafi
  • Muhammad Shahbaz
  • Khan Kashif ur Rehman
  • Khalid Javed Iqbal
  • Kashif Akram
  • Irfan Baboo
  • Shaukat Hussain Munawar
  • Muhammad Mazhar Munir
  • Rizwana Sultan
  • Hamid Majeed
  • Ilaria Cacciotti
  • Tuba Esatbeyoglu
  • Sameh A. Korma

External Research Organisations

  • Northwest University China
  • Muhammad Nawaz Shareef University of Agriculture, Multan
  • Women University of Azad Jammu and Kashmir
  • The Islamia University of Bahawalpur
  • Cholistan University of Veterinary and Animal Sciences
  • Sapienza Università di Roma
  • Zagazig University
  • South China University of Technology
View graph of relations

Details

Original languageEnglish
Article number140
JournalFermentation
Volume9
Issue number2
Publication statusPublished - 30 Jan 2023

Abstract

The intestinal tract microbiota influences many aspects of the dietary components on colon health and during enteric infections, thus, playing a pivotal role in the colon health. Therefore, the eugenol (EU) nano-emulsion effective concentration reported in our previous study against cancer cells should be explored for safety against beneficial microbes. We evaluated the sensitivity of Bifidobacterium breve and B. adolescentis against EU-loaded nano-emulsions at 0, 300, 600 and 900 µm, which were effective against colon and liver cancer cells. Both B. breve and B. adolescentis showed comparable growth ranges to the control group at 300 and 600 µm, as evident from the plate count experimental results. However, at 900 µm, a slight growth variation was revealed with respect to the control group. The real-time inhibition determination through flow cytometry showed B. breve viable, sublethal cells (99.49 and 0.51%) and B. adolescentis (95.59 and 0.15%) at 900 µm, suggesting slight inhibition even at the highest tested concentration. Flow cytometry proved to be a suitable quantitative approach that has revealed separate live, dead, and susceptible cells upon treatment with EU nano-emulsion against Escherichia coli. Similarly, in the case of B. breve and B. adolescentis, the cells showed only live cells that qualitatively suggest EU nano-emulsion safety. To judge the viability of these sublethal populations of B. breve and B. adolescentis, Fourier transforms infrared spectroscopy was carried out, revealing no peak shift for proteins, lipids, DNA and carbohydrates at 900 µm EU nano-emulsion compared to the control. On the other hand, EU-loaded nano-emulsions (900 µm)-treated E. coli showed a clear peak shift for a membrane protein, lipids, DNA and carbohydrates. This study provides insights to utilize plant phenols as safe medicines as well as dietary supplements.

Keywords

    Bifidobacterium adolescentis, bioavailability, dietary supplements, encapsulation, eugenol, flow cytometry, infrared spectroscopy, nano-emulsion

ASJC Scopus subject areas

Sustainable Development Goals

Cite this

Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry. / Majeed, Usman; Shafi, Afshan; Shahbaz, Muhammad et al.
In: Fermentation, Vol. 9, No. 2, 140, 30.01.2023.

Research output: Contribution to journalArticleResearchpeer review

Majeed, U, Shafi, A, Shahbaz, M, Rehman, KKU, Iqbal, KJ, Akram, K, Baboo, I, Munawar, SH, Munir, MM, Sultan, R, Majeed, H, Cacciotti, I, Esatbeyoglu, T & Korma, SA 2023, 'Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry', Fermentation, vol. 9, no. 2, 140. https://doi.org/10.3390/fermentation9020140
Majeed, U., Shafi, A., Shahbaz, M., Rehman, K. K. U., Iqbal, K. J., Akram, K., Baboo, I., Munawar, S. H., Munir, M. M., Sultan, R., Majeed, H., Cacciotti, I., Esatbeyoglu, T., & Korma, S. A. (2023). Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry. Fermentation, 9(2), Article 140. https://doi.org/10.3390/fermentation9020140
Majeed U, Shafi A, Shahbaz M, Rehman KKU, Iqbal KJ, Akram K et al. Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry. Fermentation. 2023 Jan 30;9(2):140. doi: 10.3390/fermentation9020140
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title = "Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial Bifidobacterium sp. along with Resistant Escherichia coli Using Flow Cytometry",
abstract = "The intestinal tract microbiota influences many aspects of the dietary components on colon health and during enteric infections, thus, playing a pivotal role in the colon health. Therefore, the eugenol (EU) nano-emulsion effective concentration reported in our previous study against cancer cells should be explored for safety against beneficial microbes. We evaluated the sensitivity of Bifidobacterium breve and B. adolescentis against EU-loaded nano-emulsions at 0, 300, 600 and 900 µm, which were effective against colon and liver cancer cells. Both B. breve and B. adolescentis showed comparable growth ranges to the control group at 300 and 600 µm, as evident from the plate count experimental results. However, at 900 µm, a slight growth variation was revealed with respect to the control group. The real-time inhibition determination through flow cytometry showed B. breve viable, sublethal cells (99.49 and 0.51%) and B. adolescentis (95.59 and 0.15%) at 900 µm, suggesting slight inhibition even at the highest tested concentration. Flow cytometry proved to be a suitable quantitative approach that has revealed separate live, dead, and susceptible cells upon treatment with EU nano-emulsion against Escherichia coli. Similarly, in the case of B. breve and B. adolescentis, the cells showed only live cells that qualitatively suggest EU nano-emulsion safety. To judge the viability of these sublethal populations of B. breve and B. adolescentis, Fourier transforms infrared spectroscopy was carried out, revealing no peak shift for proteins, lipids, DNA and carbohydrates at 900 µm EU nano-emulsion compared to the control. On the other hand, EU-loaded nano-emulsions (900 µm)-treated E. coli showed a clear peak shift for a membrane protein, lipids, DNA and carbohydrates. This study provides insights to utilize plant phenols as safe medicines as well as dietary supplements.",
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AU - Majeed, Usman

AU - Shafi, Afshan

AU - Shahbaz, Muhammad

AU - Rehman, Khan Kashif ur

AU - Iqbal, Khalid Javed

AU - Akram, Kashif

AU - Baboo, Irfan

AU - Munawar, Shaukat Hussain

AU - Munir, Muhammad Mazhar

AU - Sultan, Rizwana

AU - Majeed, Hamid

AU - Cacciotti, Ilaria

AU - Esatbeyoglu, Tuba

AU - Korma, Sameh A.

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Y1 - 2023/1/30

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