PMID- 33933521
OWN - NLM
STAT- MEDLINE
DCOM- 20211214
LR  - 20211214
IS  - 1872-8359 (Electronic)
IS  - 0167-7012 (Linking)
VI  - 185
DP  - 2021 Jun
TI  - Assessment of stool collection and storage conditions for in vitro human gut 
      model studies.
PG  - 106230
LID - S0167-7012(21)00098-1 [pii]
LID - 10.1016/j.mimet.2021.106230 [doi]
AB  - BACKGROUND: The role of the gut microbiota in health and disease is becoming 
      increasingly apparent. Faeces is the most accessible sample to collect from human 
      volunteers for studying the gut microbiota. However, the impact of stool 
      collection and storage conditions on microbial and metabolic profiles have not 
      been fully evaluated. By understanding the effect of different stool collection 
      and storage conditions on microbial and metabolic composition, we can consider 
      these parameters in the design of in vitro fermentation studies. METHODS: Stool 
      samples from 3 volunteers were stored under 5 different conditions to mimic 
      methods that researchers may use to collect and store stool samples for study of 
      the gut microbiota, including: fresh sample used within 10 min; stored on wet ice 
      (4  degrees C) for 60 min; stored in an anaerobic chamber in a temperature-controlled bag 
      (4  degrees C) for 60 min; freezing at -20  degrees C for 60 min and freezing at -20  degrees C for 
      60 min and then at -80  degrees C for 2 weeks. The stored samples were added to basal 
      medium in batch culture fermenters alone (negative control) or with 5 g 
      2'-Fucosyllactose (2'FL) Human Milk Oligosaccharide (HMO) (as a positive 
      fermentation control). Samples were collected at 3 timepoints (0, 12 and 24 h) 
      for analysis by Flow Cytometry-Fluorescent In Situ Hybridisation (FC-FISH) and 
      (1)H-Nuclear Magnetic Resonance (NMR) spectroscopy to assess the impact on 
      microbial and metabolic profiles, respectively. RESULTS: Freezing stool 
      significantly impacted microbial numbers and activity during in vitro 
      fermentations, whereas storing the stool on wet ice (4  degrees C) or in an anaerobic 
      chamber at 4  degrees C for 60 min had minimal effects on microbial and metabolic 
      profiles throughout the 24 h batch culture fermentation experiments. DISCUSSION: 
      For in vitro batch culture fermentation studies where it may not be practical or 
      possible to use fresh stool, either storing the stool on wet ice (4  degrees C) or in an 
      anaerobic chamber at 4  degrees C for 60 min could be plausible alternatives to maintain 
      microbial and metabolic profiles for analysis.
CI  - Copyright (c) 2021 Elsevier B.V. All rights reserved.
FAU - Newland, Grace A I
AU  - Newland GAI
AD  - Food Microbial Sciences Unit, Department of Food and Nutritional Sciences, 
      University of Reading, Reading RG6 6AP, UK.
FAU - Gibson, Glenn R
AU  - Gibson GR
AD  - Food Microbial Sciences Unit, Department of Food and Nutritional Sciences, 
      University of Reading, Reading RG6 6AP, UK.
FAU - Jackson, Frances L
AU  - Jackson FL
AD  - Food Microbial Sciences Unit, Department of Food and Nutritional Sciences, 
      University of Reading, Reading RG6 6AP, UK.
FAU - Wijeyesekera, Anisha
AU  - Wijeyesekera A
AD  - Food Microbial Sciences Unit, Department of Food and Nutritional Sciences, 
      University of Reading, Reading RG6 6AP, UK. Electronic address: 
      a.wijeyesekera@reading.ac.uk.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20210429
PL  - Netherlands
TA  - J Microbiol Methods
JT  - Journal of microbiological methods
JID - 8306883
SB  - IM
MH  - Adult
MH  - Batch Cell Culture Techniques/methods
MH  - Feces/*microbiology
MH  - Female
MH  - Fermentation
MH  - Flow Cytometry/methods
MH  - Freezing
MH  - *Gastrointestinal Microbiome
MH  - Humans
MH  - In Situ Hybridization, Fluorescence/methods
MH  - Magnetic Resonance Spectroscopy/methods
MH  - Metabolome
MH  - Milk, Human
MH  - Specimen Handling/*methods
MH  - Temperature
OTO - NOTNLM
OT  - (1)H NMR spectroscopy
OT  - Flow cytometry-fluorescent in situ hybridisation (FC-FISH)
OT  - Gut metabolome
OT  - Gut microbiota
OT  - In vitro batch cultures
OT  - Stool storage
EDAT- 2021/05/03 06:00
MHDA- 2021/12/15 06:00
CRDT- 2021/05/02 20:43
PHST- 2021/03/15 00:00 [received]
PHST- 2021/04/20 00:00 [revised]
PHST- 2021/04/20 00:00 [accepted]
PHST- 2021/05/03 06:00 [pubmed]
PHST- 2021/12/15 06:00 [medline]
PHST- 2021/05/02 20:43 [entrez]
AID - S0167-7012(21)00098-1 [pii]
AID - 10.1016/j.mimet.2021.106230 [doi]
PST - ppublish
SO  - J Microbiol Methods. 2021 Jun;185:106230. doi: 10.1016/j.mimet.2021.106230. Epub 
      2021 Apr 29.