Applied Microbiology and Biotechnology

, Volume 102, Issue 24, pp 10675–10690 | Cite as

E. coli Nissle microencapsulation in alginate-chitosan nanoparticles and its effect on Campylobacter jejuni in vitro

  • Asmaa Mawad
  • Yosra A. Helmy
  • Abdel-Gawad Shalkami
  • Dipak Kathayat
  • Gireesh RajashekaraEmail author
Applied microbial and cell physiology


Microencapsulation enhances the oral delivery of probiotic bacteria. In this study, the probiotic Escherichia coli Nissle 1917 (EcN) was microencapsulated using alginate and chitosan nanoparticles. The result showed 90% encapsulation yield of EcN, and the encapsulated EcN displayed significantly (P < 0.05) increased survival in low pH (1.5), high bile salt concentration (4%), and high temperature (70 °C). The most effective cryopreservatives of EcN during freezing and thawing was skim milk and sucrose. Exposure to microencapsulated EcN significantly (P < 0.05) reduced the Campylobacter jejuni growth by 2 log CFU. The rate of EcN release from microcapsule was 9.2 × 105 cell min−1, and the appropriate model to describe its release kinetics was zero order. Importantly, the entrapment of EcN inside the microcapsule did not eliminate the exterior diffusion of EcN produced antioxidant compounds. In addition, the EcN microcapsule efficiently adhered to intestinal HT-29 cells and the pre-treatment of HT-29 cells with EcN-microcapsule for 4 h significantly (P < 0.05) reduced the invasion (1.9 log) of C. jejuni; whereas, completely abolished the intracellular survival. Furthermore, HT-29 cells pre-treated with encapsulated EcN in PCR array showed decreased expression (> 1.5-fold) of genes encoding chemokines, toll-like receptors, interleukins, and tumor necrosis factors. In conclusion, the alginate-chitosan microcapsule can provide effectual platform to deliver probiotic EcN and thereby can reduce the Campylobacter infection in chickens and humans.


EcN Camplyobacter Microcapsulation Alginate Chitosan 



We thank Dr. Tea Meulia, Molecular and Cellular Imaging Center (, Ohio Agricultural Research and Development Centrer (OARDC), The Ohio State University for providing assistance with confocal microscopy analyses.

Funding information

Research in Dr. Rajashekara’s laboratory was supported by funding from National Institute for Food and Agriculture (NIFA), U.S. Department of Agriculture, National Institute of Health, and Federal and State funds appropriated to the OARDC. Dr. Asmaa Mawad was supported by the Egyptian Ministry of Higher Education.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interest.

Ethical approval

This article does not contain any studies with human participants or animals.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Asmaa Mawad
    • 1
    • 2
  • Yosra A. Helmy
    • 1
    • 3
  • Abdel-Gawad Shalkami
    • 1
    • 4
  • Dipak Kathayat
    • 1
  • Gireesh Rajashekara
    • 1
    Email author
  1. 1.Food Animal Health Research Program, Department of Veterinary Preventive Medicine, Ohio Agricultural Research and Development CenterThe Ohio State UniversityWoosterUSA
  2. 2.Botany and Microbiology Department, Faculty of ScienceAssiut UniversityAssiutEgypt
  3. 3.Department of Animal Hygiene, Zoonoses and Animal Ethology, Faculty of Veterinary MedicineSuez Canal UniversityIsmailiaEgypt
  4. 4.Pharmacology and Toxicology Department, Faculty of PharmacyAl-Azhar University, Assiut, BranchAssiutEgypt

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