Assessment of Growth Inhibition of Eugenol-Loaded Nano-Emulsions against Beneficial <i>Bifidobacterium</i> sp. along with Resistant <i>Escherichia coli</i> Using Flow Cytometry
Usman Majeed,
Afshan Shafi,
Muhammad Shahbaz,
Kashif ur Rehman Khan,
Khalid Javed Iqbal,
Kashif Akram,
Irfan Baboo,
Shaukat Hussain Munawar,
Muhammad Mazhar Munir,
Rizwana Sultan,
Hamid Majeed,
Ilaria Cacciotti,
Tuba Esatbeyoglu,
Sameh A. Korma
Affiliations
Usman Majeed
College of Food Science and Technology, Northwest University, Xi’an 710069, China
Afshan Shafi
Department of Food Science and Technology, Mian Nawaz Sharif Agriculture University, Multan 60000, Pakistan
Muhammad Shahbaz
Department of Zoology, Women University of Azad Jammu and Kashmir, Bagh 12500, Pakistan
Kashif ur Rehman Khan
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan
Khalid Javed Iqbal
Department of Zoology, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan
Kashif Akram
Department of Food Science and Technology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Irfan Baboo
Department of Zoology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Shaukat Hussain Munawar
Department of Pharmacology and Toxicology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Muhammad Mazhar Munir
Department of Pharmacology and Toxicology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Rizwana Sultan
Department of Pathology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Hamid Majeed
Department of Food Science and Technology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur 63100, Pakistan
Ilaria Cacciotti
Department of Engineering, INSTM RU, University of Rome “Niccolò Cusano”, Via Don Carlo Gnocchi 3, 00166 Roma, Italy
Tuba Esatbeyoglu
Department of Food Development and Food Quality, Institute of Food Science and Human Nutrition, Gottfried Wilhelm Leibniz University Hannover, Am Kleinen Felde 30, 30167 Hannover, Germany
Sameh A. Korma
Department of Food Science, Faculty of Agriculture, Zagazig University, Zagazig 44519, Egypt
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.
