Advertisement

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

Abstract

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.

Keywords

Nano-TiO2 Nano-silver Chocolate, Probiotics 

Notes

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)

References

  1. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215(3):403–410.  https://doi.org/10.1016/s0022-2836(05)80360-2 CrossRefGoogle Scholar
  2. Amara AA, Shibl A (2015) Role of Probiotics in health improvement, infection control and disease treatment and management. Saudi Pharm J 23(2):107–114CrossRefGoogle Scholar
  3. Bauer AW, Kirby WMM, Sherris JC, Turck M (1966) Antibiotic Susceptibility Testing by a Standardized Single Disk Method. American Journal of Clinical Pathology 45 (4_ts):493–496Google Scholar
  4. Brosius J, Palmer ML, Kennedy PJ, Noller HF (1978) Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proceedings of the National Academy of Science, USA 75:4801–4805Google Scholar
  5. Cappuccino JG, Sherman N (1996) Microbiology: a laboratory manual. Benjamin/Cummings Publishing Company, San FranciscoGoogle Scholar
  6. Chamley CA (2005) Developmental anatomy and physiology of children: a practical approach. Elsevier/Churchill Livingstone, LondonGoogle Scholar
  7. Clemente JC, Ursell LK, Parfrey LW, Knight R (2012) The impact of the gut microbiota on human health: an integrative view. Cell 148(6):1258–1270CrossRefGoogle Scholar
  8. Del Nobile MA, Lucera A, Costa C, Conte A (2012) Food applications of natural antimicrobial compounds. Front Microbiol 3(287).  https://doi.org/10.3389/fmicb.2012.00287
  9. Dunne C, O’Mahony L, Murphy L, Thornton G, Morrissey D, O’Halloran S, Feeney M, Flynn S, Fitzgerald G, Daly C, Kiely B, O’Sullivan GC, Shanahan F, Collins JK (2001) In vitro selection criteria for probiotic bacteria of human origin: correlation with in vivo findings. Am J Clin Nutr 73(2 Suppl):386 s–392 s.  https://doi.org/10.1093/ajcn/73.2.386s CrossRefGoogle Scholar
  10. Edwards PR (1972) In: Edwards PR, Ewing WH (eds) Identification of Enterobacteriaceae. Burgess Pub. Co, MinneapolisGoogle Scholar
  11. FAO (1993)Small-Scale Dairy Farming Manual Volume 1Google Scholar
  12. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39(4):783–791CrossRefGoogle Scholar
  13. Fuller R (1991) Probiotics in human medicine. Gut 32(4):439–442CrossRefGoogle Scholar
  14. Guandalini S (2011) Probiotics for prevention and treatment of diarrhea. J Clin Gastroenterol 45 Suppl:S149–S153.  https://doi.org/10.1097/MCG.0b013e3182257e98 CrossRefGoogle Scholar
  15. Hajela EA (2014) Probiotic foods: can their increasing use in India ameliorate the burden of chronic lifestyle disorders? The Indian. J Med Res 139(1):19–26Google Scholar
  16. Hart ML, Meyer A, Johnson PJ, Ericsson AC (2015) Comparative evaluation of DNA extraction methods from feces of multiple host species for downstream next-generation sequencing. PLoS One 10(11):e0143334–e0143334.  https://doi.org/10.1371/journal.pone.0143334 CrossRefGoogle Scholar
  17. Heinrich U, Fuhst R, Rittinghausen S, Creutzenberg O, Bellmann B, Koch W, Levsen K (1995) chronic inhalation exposure of Wistar rats and two different strains of mice to diesel engine exhaust, carbon black, and titanium dioxide. Inhal Toxicol 7(4):533–556CrossRefGoogle Scholar
  18. Holzapfel WH, Haberer P, Geisen R, Björkroth J, Schillinger U (2001) Taxonomy and important features of probiotic microorganisms in food and nutrition. Am J Clin Nutr 73(2):365s–373sCrossRefGoogle Scholar
  19. Khan ST, Malik A (2019) Engineered nanomaterials for water decontamination and purification: from lab to products. J Hazard Mater 5(363):295–308.  https://doi.org/10.1016/j.jhazmat.2018.09.091 CrossRefGoogle Scholar
  20. Khan ST, Tamura T, Takagi M, Shin-ya K (2010)Streptomyces tateyamensis sp. nov., Streptomyces marinus sp. nov. and Streptomyces haliclonae sp. nov., isolated from the marine sponge Haliclona sp. Int J Syst Evol Microbiol 60(12):2775–2779.  https://doi.org/10.1099/ijs.0.019869-0 CrossRefGoogle Scholar
  21. Khan ST, Al-Khedhairy AA, Musarrat J (2015) ZnO and TiO2 nanoparticles as novel antimicrobial agents for oral hygiene: a review. J Nanopart Res 17(6):276.  https://doi.org/10.1007/s11051-015-3074-6
  22. Khan ST, Musarrat J, Al-Khedhairy AA (2016) Countering drug resistance, infectious diseases, and sepsis using metal and metal oxides nanoparticles: current status. Colloids Surf B: Biointerfaces 146:70–83.  https://doi.org/10.1016/j.colsurfb.2016.05.046 CrossRefGoogle Scholar
  23. Khan ST, Ahmad J, Ahamed M, Jousset A (2018)Sub-lethal doses of widespread nanoparticles promote antifungal activity in Pseudomonas protegens CHA0. Sci Total Environ 627:658–662CrossRefGoogle Scholar
  24. Macfarlane GT, Cummings JH (1999) Probiotics and prebiotics: can regulating the activities of intestinal bacteria benefit health? BMJ 318(7189):999–1003CrossRefGoogle Scholar
  25. Manero A, Blanch AR (1999) Identification of Enterococcus spp. with a biochemical key. Appl Environ Microbiol 65(10):4425–4430Google Scholar
  26. McFarland LV (2015) From yaks to yogurt: the history, development, and current use of probiotics. Clin Infect Dis 60(Suppl 2):S85–S90.  https://doi.org/10.1093/cid/civ054 CrossRefGoogle Scholar
  27. Merritt JH, Kadouri DE, O’Toole GA (2005) Growing and analyzing static biofilms. Curr Protoc Microbiol Chapter 1:Unit-1B.1.  https://doi.org/10.1002/9780471729259.mc01b01s00 Google Scholar
  28. Mshana RN, Tadesse G, Abate G, Miörner H (1998) Use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide for rapid detection of rifampin-resistantMycobacterium tuberculosis. J Clin Microbiol 36(5):1214–1219Google Scholar
  29. O’Toole GA (2011) Microtiter dish biofilm formation assay. J Vis Exp 47.  https://doi.org/10.3791/2437
  30. Patterson AL (1939) The diffraction of X-rays by small crystalline particles. Phys Rev 56(10):972–977.  https://doi.org/10.1103/PhysRev.56.972 CrossRefGoogle Scholar
  31. Reid G (2015) The growth potential for dairy probiotics. Int Dairy J 49:16–22CrossRefGoogle Scholar
  32. Sadeghi R, Rodriguez RJ, Yao Y, Kokini JL (2017) Advances in nanotechnology as they pertain to food and agriculture: benefits and risks. Annu Rev Food Sci Technol 8(1):467–492CrossRefGoogle Scholar
  33. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4(4):406–425Google Scholar
  34. Sarles WB, Hammer BW (1932) Observations on Bacillus coagulans. J Bacteriol 23(4):301–314Google Scholar
  35. Schleifer KH, Kilpper-Bälz R (1984) Transfer of Streptococcus faecalis and Streptococcus faecium to the Genus Enterococcus nom. rev. as Enterococcus faecalis comb. nov. and Enterococcus faecium comb. nov. 34(1):31–34Google Scholar
  36. Shi H, Magaye R, Castranova V, Zhao J (2013) Titanium dioxide nanoparticles: a review of current toxicological data. Part Fibre Toxicol 10:15–15CrossRefGoogle Scholar
  37. Spor A, Koren O, Ley R (2011) Unravelling the effects of the environment and host genotype on the gut microbiome. Nat Rev Microbiol 9(4):279–290CrossRefGoogle Scholar
  38. Valdes AM, Walter J, Segal E, Spector TD (2018) Role of the gut microbiota in nutrition and health. BMJ 361. https://doi.org/10.1136/bmj.k2179%JGoogle Scholar
  39. Verna EC, Lucak S (2010) Use of probiotics in gastrointestinal disorders: what to recommend? Ther Adv Gastroenterol 3(5):307–319.  https://doi.org/10.1177/1756283X10373814 CrossRefGoogle Scholar
  40. Weir A, Westerhoff P, Fabricius L, Hristovski K, von Goetz N (2012) Titanium dioxide nanoparticles in food and personal care products. Environ Sci Technol Lett 46(4):2242–2250.  https://doi.org/10.1021/es204168d CrossRefGoogle Scholar
  41. Yang Y-W, Chen M-K, Yang B-Y, Huang X-J, Zhang X-R, He L-Q, Zhang J, Hua Z-C(2015) Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in mouse feces. Appl Environ Microbiol 81(19):6749–6756CrossRefGoogle Scholar
  42. Zuo T, Kamm MA, Colombel J-F, Ng SC (2018) Urbanization and the gut microbiota in health and inflammatory bowel disease. Nat Rev Gastroenterol Hepatol 15(7):440–452CrossRefGoogle Scholar

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

Personalised recommendations