Optimisation of inoculum concentration and incubation duration for an in vitro hindgut dry matter digestibility assay

Coles, Leah; Moughan, Paul J.; Awati, A.; Darragh, Alison J.

doi:10.1016/j.foodchem.2012.08.045

Cited by 19 publications

(31 citation statements)

References 27 publications

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“…The pig ileal digesta are incubated (in vitro) with a pooled human fecal inoculum to model hindgut fermentation in humans. The human fecal inoculum is used to model the microbiota present in the hindgut (15,(19)(20)(21)(22). This approach provides information on the SCFAs entering the hindgut, fermentable organic matter (OM) entering the hindgut, OM hindgut fermentability, estimates of hindgut SCFA production, SCFAs exiting the hindgut, and estimates of total hindgut SCFA absorption (19) and has been shown to give accurate data (15).…”

Section: Introductionmentioning

confidence: 99%

Development of an In Vivo and In Vitro Ileal Fermentation Method in a Growing Pig Model

Montoya

Haas

Moughan

2018

The Journal of Nutrition

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Section: Introductionmentioning

confidence: 99%

Development of an In Vivo and In Vitro Ileal Fermentation Method in a Growing Pig Model

Montoya

Haas

Moughan

2018

The Journal of Nutrition

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“…To measure SCFA production by gut microbiota, we adapted the in vitro approach of Edwards et al 1996 (41). This method was specifically designed to study fermentation of starch in the human lower GI tract, and has since been used to measure metabolite production from human stool samples when exposed to prebiotic fiber (42–44). In brief, we homogenized previously frozen feces in reduced phosphate buffered saline (pH 7.0 ± 0.1) to create a fecal slurry with a final concentration of 100g/L (Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…We chose our culture conditions, namely a media-free approach that does not add any nutrients beyond what is present in the stool, in an effort to avoid inducing artificial selective conditions within our cultures. Prior experimental digestion studies have shown that prebiotic response patterns can be recapitulated across varying culture conditions (42, 44). Indeed, we found strong correlation in SCFA production between cultures grown with our media-free approach and those grown in a more conventional medium containing added nitrogen, vitamins, minerals, and acetate.…”

Section: Discussionmentioning

confidence: 99%

“…Aliquoted stool was thawed at room temperature in an anaerobic chamber. Once thawed, stool was weighed and placed into a polyethylene filter bag with 0.33mm pore size (Whirl-Pak B01385) and 10mL of anaerobic 1X PBS was added for each gram of stool, resulting in a 10% w/v fecal slurry, similar to previous studies (41, 42, 70, 71). During our validation experiments, a medium designed to simulate colonic contents was used in place of 1X PBS to create stool slurries (46).…”

Section: Methodsmentioning

confidence: 99%

“…The resulting fermentation conditions where therefore 5% fecal slurry with 0.5% prebiotic (w/v). A 5% fecal slurry was selected because its fermentative capacity has been previously demonstrated to be insensitive to small variations in concentration and is feasible to work with using this method (42). A 0.5% final concentration of prebiotic in the context of a 5% fecal slurry is analogous to an average adult consuming 20g of dietary fiber per day, assuming an average daily stool mass of 200g (73).…”

Section: Methodsmentioning

confidence: 99%

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Short-chain fatty acid production by gut microbiota from children with obesity is linked to bacterial community composition and prebiotic choice

Holmes

Silverman

Dressman

et al. 2020

Preprint

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23Pediatric obesity remains a public health burden and continues to increase in 24 prevalence. The gut microbiota plays a causal role in obesity and is a promising 25 therapeutic target. Specifically, the microbial production of short-chain fatty acids 26 (SCFA) from the fermentation of otherwise indigestible dietary carbohydrates may 27 protect against pediatric obesity and metabolic syndrome. Still, it has not been 28 demonstrated that therapies involving microbiota-targeting carbohydrates, known as 29 prebiotics, will enhance gut bacterial SCFA production in children and adolescents with 30 obesity (age 10-18). Here, we used an in vitro system to examine the SCFA production 31 by fecal microbiota from 17 children with obesity when exposed to five different 32 commercially available over-the-counter (OTC) prebiotic supplements. We found 33 microbiota from all 17 patients actively metabolized most prebiotics. Still, supplements 34 varied in their acidogenic potential. Significant inter-donor variation also existed in SCFA 35 production, which 16S rRNA sequencing supported as being associated with differences 36 in the host microbiota composition. Last, we found that neither fecal SCFA 37 concentration, microbiota SCFA production capacity, nor markers of obesity positively 38 correlated with one another. Together, these in vitro findings suggest the hypothesis that 39 OTC prebiotic supplements may be unequal in their ability to stimulate SCFA production 40 in children and adolescents with obesity, and that the most acidogenic prebiotic may 41 differ across individuals. 42 43 44 IMPORTANCE Pediatric obesity remains a major public health problem in the US, where 45 17% of children and adolescents are obese, and rates of pediatric 'severe obesity' are 46 increasing. Children and adolescents with obesity face higher health risks, and non-47 invasive therapies for pediatric obesity often have limited success. The human gut 48 microbiome has been implicated in adult obesity, and microbiota-directed therapies can 49 aid weight loss in adults with obesity. However, less is known about the microbiome in 50 pediatric obesity, and microbiota-directed therapies are understudied in children and 51 adolescents. Our research has two important findings: 1) dietary prebiotics (fiber) cause 52 the microbiota from adolescents with obesity to produce more SCFA, and 2) the 53 effectiveness of each prebiotic is donor-dependent. Together, these findings suggest 54 that prebiotic supplements could help children and adolescents with obesity, but that 55 these therapies may not be one-size-fits-all. 56 developed long-term complications, we pursued three specific lines of inquiry: 1) whether 132 different types of prebiotics lead to differences in SCFA production by gut microbiota from 133 adolescents with obesity; 2) whether the effects of prebiotics are shaped by inter-134 individual differences in gut microbiota structure; and, 3) whether fecal SCFA production 135 is associated with protection from obesity. 136

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Validation of a dual in vivo–in vitro assay for predicting the digestibility of nutrients in humans

et al. 2013

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Optimisation of inoculum concentration and incubation duration for an in vitro hindgut dry matter digestibility assay

Cited by 19 publications

References 27 publications

Development of an In Vivo and In Vitro Ileal Fermentation Method in a Growing Pig Model

Development of an In Vivo and In Vitro Ileal Fermentation Method in a Growing Pig Model

Short-chain fatty acid production by gut microbiota from children with obesity is linked to bacterial community composition and prebiotic choice

Validation of a dual in vivo–in vitro assay for predicting the digestibility of nutrients in humans

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