Antibacterial and Antibiofilm Activity of Eucalyptus camaldulensis Derived Fe3O4 Nano-particles against Foodborne Pathogens

  • Afshan Zia Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan
  • Sadia Tahir Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan
  • Saba Saba Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan
  • Abid Hussain Department of Microbiology and Molecular Genetics, University of Okara, Okara, Pakistan
  • Iqra Arooj Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan
  • Laraib Aslam Department of Microbiology and Molecular Genetics, The Women University Multan, Multan, Pakistan
Keywords: Foodborne pathogen, Eucalyptus camaldulensis extract, Nano-particles


Foodborne pathogens are zoonotic and multidrug resistant, which are not only affecting economy but also accountable for public health burdens. The present study was aimed to evaluate the efficacy of medicinal plant i.e., Eucalyptus camaldulensis extract mediated Fe3O4 and ZnO nanoparticles against the 27 food-borne pathogenic strains, isolated from milk, meat, dry fruits and vegetable samples collected from Multan, Pakistan. In phytochemical screening, the plant extract was found to contain numerous bioactive compounds including flavonoids, phenolic compounds, tannins, quinones, and anthocyanins. Fe3O4 NPs synthesized from Eucalyptus camaldulensis displayed the highest antibacterial activity with zones of inhibition of 12-13 mm against pathogens. Fe3O4 NPs were found to have highest anti-inflammatory potential with recorded percentage of 67% at 40 µg/ml. Fe3O4 NPs also demonstrated the highest antibiofilm activity after 120 hours of incubation. For DDPH antioxidant assay, the highest antioxidant activity was displayed by Fe3O4 NPs and their absorbance recorded was 1.43. Therefore, Eucalyptus camaldulensis mediated Fe3O4 NPs proved as an effective and eco-friendly approach to combat multidrug resistance in bacterial infections through characteristic antibacterial, antibiofilm and antioxidant properties.


  1. Aladhadh M (2023). A Review of Modern Methods for the Detection of Foodborne Pathogens. Microorg. 11: 1111-1130.
  2. Alves de Aguiar Bernardo Y, KaicAlves do Rosario D, Adam Conte-Junior C (2021). Ultrasound on Milk Decontamination: Potential and Limitations against Foodborne Pathogens and Spoilage Bacteria. Food Rev. Int. 39: 320-333.
  3. Andrade-Zavaleta K, Chacon-Laiza Y, Asmat-Campos D, Raquel-Checca N (2022). Green Synthesis of Superparamagnetic Iron Oxide Nanoparticles with Eucalyptus globulus Extract and Their Application in the Removal of Heavy Metals from Agricultural Soil. Molecules. 27: 1367-1384.
  4. Barzinjy AA, Azeez HH (2020). Green synthesis and characterization of zinc oxide nanoparticles using Eucalyptus globulus Labill. leaf extract and zinc nitrate hexahydrate salt. SN Appl. Sci. 2: 991-1004.
  5. Cappuccino J, Welsh C, 2019. Microbiology: A Laboratory Manual, Pearson.
  6. Castro-Vargas RE, Herrera-Sanchez MP, Rodriguez-Hernandez R, Rondon-Barragan IS (2020). Antibiotic resistance in Salmonella spp. isolated from poultry: A global overview. Vet. World. 13: 2070-2084.
  7. Chandrakala V, Aruna V, Angajala G (2022). Review on metal nanoparticles as nanocarriers: current challenges and perspectives in drug delivery systems. Emergent. Mater. 5: 1593-1615.
  8. Dela Cruz TEE, Torres JMO (2012). Gelatin hydrolysis test protocol. Am. Soc. Microbiol. 1-10.
  9. Dutt Y, Dhiman R, Singh T, Vibhuti A, Gupta A, Pandey RP, Raj VS, Chang C-M, Priyadarshini A (2022). The Association between Biofilm Formation and Antimicrobial Resistance with Possible Ingenious Bio-Remedial Approaches. Antibiotics. 11: 930-955.
  10. Elbehiry A, Abalkhail A, Marzouk E, Elmanssury AE, Almuzaini AM, Alfheeaid H, Alshahrani MT, Huraysh N, Ibrahem M, Alzaben F, Alanazi F, Alzaben M, Anagreyyah SA, Bayameen AM, Draz A, Abu-Okail A (2023). An Overview of the Public Health Challenges in Diagnosing and Controlling Human Foodborne Pathogens. Vaccines (Basel). 11: 725-769
  11. Kalishwaralal K, BarathManiKanth S, Pandian SR, Deepak V, Gurunathan S (2010). Silver nanoparticles impede the biofilm formation by Pseudomonas aeruginosa and Staphylococcus epidermidis. Colloids Surf B Biointerfaces. 79: 340-344.
  12. Liu X, Yao H, Zhao X, Ge C (2023). Biofilm Formation and Control of Foodborne Pathogenic Bacteria. Molecules. 28: 2432-2451.
  13. Mann CM, Markham JL (1998). A new method for determining the minimum inhibitory concentration of essential oils. J Appl. Microbiol. 84: 538-544.
  14. Mathur T, Singhal S, Khan S, Upadhyay DJ, Fatma T, Rattan A (2006). Detection of Biofilm Formation among the Clinical Isolates of Staphylococci: An Evaluation of Three Different Screening Methods. Indian J. Med. Microbiol. 24: 25-29.
  15. Mohamad F, Alzahrani RR, Alsaadi A, Alrfaei BM, Yassin AEB, Alkhulaifi MM, Halwani M (2023). An Explorative Review on Advanced Approaches to Overcome Bacterial Resistance by Curbing Bacterial Biofilm Formation. Infect. Drug Resist. 16: 19-49.
  16. Nur MA, Uddin MR, Meghla NS, Uddin MJ, Amin MZ (2023). In vitro anti-oxidant, anti-inflammatory, anti-bacterial, and cytotoxic effects of extracted colorants from two species of dragon fruit (Hylocereus spp.). Food Chem. Adv. 2: 100318-100327.
  17. Pushpalatha C, Suresh J, Gayathri VS, Sowmya SV, Augustine D, Alamoudi A, Zidane B, Mohammad Albar NH, Patil S (2022). Zinc Oxide Nanoparticles: A Review on Its Applications in Dentistry. Front Biotechnol. Bioeng. 10: 917990-917996.
  18. Reygaert WC (2018). An overview of the antimicrobial resistance mechanisms of bacteria. AIMS Microbiol. 4: 482-501.
  19. Ronavari A, Igaz N, Adamecz DI, Szerencses B, Molnar C, Konya Z, Pfeiffer I, Kiricsi M (2021). Green Silver and Gold Nanoparticles: Biological Synthesis Approaches and Potentials for Biomedical Applications. Molecules. 26: 844-893..
  20. Schlesier K, Harwat M, Bohm V, Bitsch R (2002). Assessment of antioxidant activity by using different in vitro methods. Free Radic. Res. 36: 177-187.
  21. Shaikh JR, Patil MK (2020). Qualitative tests for preliminary phytochemical screening: An overview. Int. J. Chem. Stud. 8: 603-608.
  22. Sharma OP, Bhat TK (2009). DPPH antioxidant assay revisited. Food Chem. 113: 1202-1205.
  23. Sheng L, Wang L (2023). Approaches for a more microbiologically and chemically safe dried fruit supply chain. Curr. Opin. Biotechnol. 80: 102912-102920.
  24. Surbhi, Kumar A, Singh S, Kumari P, Rasane P (2021). Eucalyptus: phytochemical composition, extraction methods and food and medicinal applications. Adv. Trad. Med. 23: 369-380.
  25. Tan HL, Lim YC, Ng LY, Lim YP (2023). Plant-mediated synthesis of iron nanoparticles for environmental application: Mini review. Mater. Today Proc. 87: 64-69.
  26. Tarabees R, Elsayed MSA, Shawish R, Basiouni S, Shehata AA (2017). Isolation and characterization of Salmonella Enteritidis and Salmonella Typhimurium from chicken meat in Egypt. J. Infect. Dev. Ctries. 11: 314-319.
  27. Villa G, Boarin M, Rosa D, Togni S, Manara DF, Bonetti L, Terzoni S (2022). Phytotherapy in urological benign disease: A systematic review. J. Urol. Nurs. 16: 174-195.
  28. Wilson C, Lukowicz R, Merchant S, Valquier-Flynn H, Caballero J, Sandoval J, Okuom M, Huber C, Brooks TD, Wilson E, Clement B, Wentworth CD, Holmes AE (2017). Quantitative and Qualitative Assessment Methods for Biofilm Growth: A Mini-review. Res. Rev. J. Eng. Technol. 6: 1-42.