TY - JOUR
T1 - Preservation of human gut microbiota inoculums for in vitro fermentations studies
AU - Carvalho, Nelson Mota de
AU - Oliveira, Diana Luazi
AU - Saleh, Mayra Anton Dib
AU - Pintado, Manuela
AU - Madureira, Ana Raquel
N1 - Funding Information:
This research was funded by Amyris Bio Products Portugal Unipessoal Lda and Escola Superior de Biotecnologia—Universidade Católica Portuguesa through Alchemy project-Capturing high value from industrial fermentation bio products (POCI-01-0247-FEDER-027578). We would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019.
Funding Information:
Funding: This research was funded by Amyris Bio Products Portugal Unipessoal Lda and Escola Superior de Biotecnologia—Universidade Católica Portuguesa through Alchemy project-Capturing high value from industrial fermentation bio products (POCI-01-0247-FEDER-027578). We would also like to thank the scientific collaboration under the FCT project UID/Multi/50016/2019.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/3
Y1 - 2021/3
N2 - The use of fecal inoculums for in vitro fermentation models requires a viable gut microbiota, capable of fermenting the unabsorbed nutrients. Fresh samples from human donors are used; however, the availability of fresh fecal inoculum and its inherent variability is often a problem. This study aimed to optimize a method of preserving pooled human fecal samples for in vitro fermentation studies. Different conditions and times of storage at −20 ◦C were tested. In vitro fermentation experiments were carried out for both fresh and frozen inoculums, and the metabolic profile compared. In comparison with the fresh, the inoculum frozen in a PBS and 30% glycerol solution, had a significantly lower (p < 0.05) bacterial count (<1 log CFU/mL). However, no significant differences (p < 0.05) were found between the metabolic profiles after 48 h. Hence, a PBS and 30% glycerol solution can be used to maintain the gut microbiota viability during storage at −20 ◦C for at least 3 months, without interfering with the normal course of colonic fermentation.
AB - The use of fecal inoculums for in vitro fermentation models requires a viable gut microbiota, capable of fermenting the unabsorbed nutrients. Fresh samples from human donors are used; however, the availability of fresh fecal inoculum and its inherent variability is often a problem. This study aimed to optimize a method of preserving pooled human fecal samples for in vitro fermentation studies. Different conditions and times of storage at −20 ◦C were tested. In vitro fermentation experiments were carried out for both fresh and frozen inoculums, and the metabolic profile compared. In comparison with the fresh, the inoculum frozen in a PBS and 30% glycerol solution, had a significantly lower (p < 0.05) bacterial count (<1 log CFU/mL). However, no significant differences (p < 0.05) were found between the metabolic profiles after 48 h. Hence, a PBS and 30% glycerol solution can be used to maintain the gut microbiota viability during storage at −20 ◦C for at least 3 months, without interfering with the normal course of colonic fermentation.
KW - Colon fermentation
KW - Glycerol
KW - Gut microbiota viability
KW - Human in vitro models
KW - Organic acids
KW - Preservation
UR - http://www.scopus.com/inward/record.url?scp=85106961632&partnerID=8YFLogxK
U2 - 10.3390/fermentation7010014
DO - 10.3390/fermentation7010014
M3 - Article
SN - 2311-5637
VL - 7
JO - Fermentation
JF - Fermentation
IS - 1
M1 - 14
ER -