Supplementation with dairy matrices impacts on homocysteine levels and gut microbiota composition of hyperhomocysteinemic mice

  • Paola Zinno
  • Vincenzo Motta
  • Barbara Guantario
  • Fausta Natella
  • Marianna Roselli
  • Cristiano Bello
  • Raffaella Comitato
  • Domenico Carminati
  • Flavio Tidona
  • Aurora Meucci
  • Paola Aiello
  • Giuditta Perozzi
  • Fabio Virgili
  • Paolo Trevisi
  • Raffaella CanaliEmail author
  • Chiara DevirgiliisEmail author
Original Contribution



Several studies highlighted a correlation between folic acid deficiency and high plasma homocysteine concentration, considered a risk factor for multifactorial diseases. Natural folates represent an emerging alternative strategy to supplementation with synthetic folic acid, whose effects are controversial. The present work was, therefore, performed in hyperhomocysteinemic mice to study the impact of supplementation with dairy matrices containing natural folates on plasma homocysteine levels and faecal microbiota composition.


Forty mice were divided into six groups, two of which fed control or folic acid deficient (FD) diets for 10 weeks. The remaining four groups were fed FD diet for the first 5 weeks and then shifted to a standard control diet containing synthetic folic acid (R) or a FD diet supplemented with folate-enriched fermented milk (FFM) produced by selected lactic acid bacteria, fermented milk (FM), or milk (M), for additional 5 weeks.


Supplementation with dairy matrices restored homocysteine levels in FD mice, although impacting differently on hepatic S-adenosyl-methionine levels. In particular, FFM restored both homocysteine and S-adenosyl-methionine levels to the control conditions, in comparison with FM and M. Next generation sequencing analysis revealed that faecal microbiota of mice supplemented with FFM, FM and M were characterised by a higher richness of bacterial species in comparison with C, FD and R groups. Analysis of beta diversity highlighted that the three dairy matrices determined specific, significant variations of faecal microbiota composition, while hyperhomocysteinemia was not associated with significant changes.


Overall, the results represent a promising starting point for the applicability of food matrices enriched in natural folates to manage hyperhomocysteinemia.


Fermented food Folate Homocysteine Microbiome 



The Authors wish to thank Kariklia Pascucci for her kind support in daily lab work, Altero Aguzzi for his help in fermented milk liophylisation and Rita Rami for the excellent care of the animals. This work was funded by the Italian Ministry of Agriculture, Food & Forestry Policies (MiPAAF), with Grant “MEDITO” (DM 12487/7303/11) and with national support to the JPI-HDHL “ENPADASI” project.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Consent for publication

All authors read and approved the final manuscript.

Supplementary material

394_2019_1911_MOESM1_ESM.docx (79 kb)
Supplementary material 1 (DOCX 79 KB)
394_2019_1911_MOESM2_ESM.docx (53 kb)
Supplementary material 2 (DOCX 52 KB)
394_2019_1911_MOESM3_ESM.tif (12.9 mb)
Supplementary material 3 Fig. S1. Per sample rarefaction curves. Each curve shows the average number of Operational Taxonomic Units (OTUs) as a function of the reads abundance subsampled at different depths. Each frame and color represent the experimental treatments, the vertical gray line indicates the lowest number of reads obtained on overall samples (n = 76187) (TIF 13183 KB)
394_2019_1911_MOESM4_ESM.xlsx (16 kb)
Supplementary material 4 (XLSX 15 KB)


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Copyright information

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

Authors and Affiliations

  1. 1.Research Centre for Food and NutritionCREA (Council for Agricultural Research and Economics)RomeItaly
  2. 2.Department of Agricultural and Food Sciences (DISTAL)University of BolognaBolognaItaly
  3. 3.Research Centre for Animal Production and AquacultureCREA (Council for Agricultural Research and Economics)LodiItaly

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