Dataset Information

An automated sensitive approach for measuring whole gut transit time.

ABSTRACT:

Background

Commonly used methods to measure whole gut transit time in rodents have yet to combine high sensitivity, objectivity, and automation. We have developed a novel method using oral gavage of non-toxic fluorescent dye particles and their detection by fluorescence imaging to enable unbiased automated detection of gut transit time simultaneously in 8 cages.

Methods

Naïve mice (n = 20) were gavaged with a non-caloric viscous suspension of 4.4% fluorescent dye in 3 groups on 2 occasions. Each group was imaged in 8 cages at 5-minute intervals using blue LEDs for illumination and a Sony full-frame mirrorless camera with a green band-pass emission filter. Custom MATLAB code counted the number of fluorescent boli per cage post hoc and provided graphical and spreadsheet output. Boli counts across a wide range of parameters were compared to blind assessments by an experimenter.

Results

Fluorescent boli were detected with high sensitivity, while unstained boli were readily rejected. All cages showed no fluorescent boli for the first ~20 frames (100 minutes), after which many cages gradually show a rise to 1-6 fluorescent boli. The mean time to first fluorescent bolus in each session was 264 ± 141 and 223 ± 81 minutes post-gavage, with no within subject consistency. There was high correlation between automated scores and that of experimenter (r = .95 ± .02), being robust to parameter changes.

Conclusions and inferences

This novel approach provides a reliable, automatic, and low-cost method of measuring gastrointestinal transit time in mice.

SUBMITTER: Baker KL

PROVIDER: S-EPMC9361480 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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Publications

An automated sensitive approach for measuring whole gut transit time.

Baker Keeley L KL Izydorczak Michelle M Jackson Ruaidhri R Verhagen Justus V JV

Neurogastroenterology and motility 20200529 9

<h4>Background</h4>Commonly used methods to measure whole gut transit time in rodents have yet to combine high sensitivity, objectivity, and automation. We have developed a novel method using oral gavage of non-toxic fluorescent dye particles and their detection by fluorescence imaging to enable unbiased automated detection of gut transit time simultaneously in 8 cages.<h4>Methods</h4>Naïve mice (n = 20) were gavaged with a non-caloric viscous suspension of 4.4% fluorescent dye in 3 groups on 2 ...[more]

PMID: 32468651

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Project description:BackgroundTransit time is an important modulator of the human gut microbiome. The inability to modify transit time as the sole variable hampers mechanistic in vivo microbiome research. We singled out gut transit time in an unprecedented in vitro approach by subjecting faecal microbial communities from six individuals with either short, medium or long in vivo transit times, to three different colonic transit times of 21, 32 and 63 h in the validated human gut in vitro model, SHIME.ResultsTransit time was identified as the single most important driver of microbial cell concentrations (52%), metabolic activity (45%) and quantitative (24%) and proportional (22%) community composition. Deceleration of transit was characterised by a significant decrease of specific Bifidobacterium and Veillonella spp. and increase of specific fibre degrading bacteria and nutrient specialists, such as Bacteroides, Prevotella, Ruminococcus, Bilophila and Akkermansia spp. These microbial communities reached a higher population density and net carbohydrate fermentation, leading to an increased SCFA production at longer transit times. In contrast, the carbohydrate-to-biomass production efficiency was increased at shorter transits, particularly in well-adapted faecal microbiomes from donors with short in vivo transit. Said adaptation was also reflected in the carbohydrate-to-SCFA conversion efficiency which varied with donor, but also colon region and SCFA chain length. A long transit time promoted propionate production, whereas butyrate production and butyrate producers were selectively enriched in the proximal colon at medium transit time.ConclusionMicrobial growth rates and nutrient utilisation efficiency mediate the species-specific gut microbiota response to in vitro transit time variation, which is the main driver of in vitro microbial load, metabolism and community composition. Given the in vivo transit time variation within and between individuals, the personalisation of in vitro transit time based on in vivo data is required to accurately study intra- and inter-individual differences in gut microbiome structure, functionality and interactions with host and environmental modulators. Video Abstract.

Dataset Information

An automated sensitive approach for measuring whole gut transit time.

Background

Methods

Results

Conclusions and inferences

Publications

An automated sensitive approach for measuring whole gut transit time.

Similar Datasets

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