Survival of probiotic bacteria in the presence of food grade nanoparticles from chocolates: an in vitro and in vivo study

  • Shams Tabrez KhanEmail author
  • Shaibi Saleem
  • Maqusood Ahamed
  • Javed Ahmad
Applied microbial and cell physiology


The use of probiotics to treat gastrointestinal diseases such as diarrhea especially in children is becoming increasingly popular. Besides, the use of nanomaterials in food products is increasing rapidly especially in candies and chocolates. How these nanomaterials influence probiotic bacteria and their activity remains unexplored. Therefore, nanomaterials from commercial chocolate were purified and characterized by using SEM–EDS and XRD. The tested chocolate contained nano-TiO2 with an average size of ~ 40 nm. The influence of the extracted TiO2 on a commercial probiotic formulation usually used to treat diarrhea in children was studied. The probiotic formulation contained Bacillus coagulans, Enterococcus faecalis, and Enterococcus faecium as evident from 16S rRNA gene sequences and polyphasic characterization. Isolated bacteria exhibited known probiotic activities like biofilm formation, acid production, growth at 6% salt, and antibiotic resistance. TiO2 from chocolates inhibited the growth and activity of the probiotic formulation over a concentration range of 125–500μg/ml in vitro. Based on results, it is estimated that 20 g of such chocolate contains enough TiO2 to disturb the gut microbial community of children aged 2–8 years with a stomach capacity of ~ 0.5–0.9 l. The in vivo study on white albino mice shows the same response but with a higher dose. The results obtained by plate counts, MTT assay, live/dead staining, and qPCR suggest that TiO2 from chocolates inhibits the growth and viability of probiotic bacteria in mice gut even at a concentration of 50–100 μg/day/mice. Therefore, TiO2 in chocolate discourages survival of probiotic bacteria in the human gut.


Nano-TiO2 Nano-silver Chocolate, Probiotics 


Funding information

The authors are grateful to the Deanship of Scientific Research, King Saud University, for funding through Vice Deanship of Scientific Research Chairs.

Compliance with ethical standards

All the experiments on the animals were performed according to the guidelines of Animal Ethics Committiee of King Saud University. This study does not contain any experiments on humans.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

253_2019_9918_MOESM1_ESM.pdf (174 kb)
ESM 1 (PDF 173 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Shams Tabrez Khan
    • 1
    Email author
  • Shaibi Saleem
    • 1
  • Maqusood Ahamed
    • 2
  • Javed Ahmad
    • 3
    • 4
  1. 1.Department of Agricultural Microbiology, Faculty of Agricultural SciencesAligarh Muslim UniversityAligarhIndia
  2. 2.King Abdullah Institute for NanotechnologyKing Saud UniversityRiyadhSaudi Arabia
  3. 3.Zoology Department, College of ScienceKing Saud UniversityRiyadhSaudi Arabia
  4. 4.Al-Jeraisy Chair for DNA ResearchKing Saud UniversityRiyadhSaudi Arabia

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