Kiwifruit-driven Microbiota, Metabolites And Implications for Human Health by Dr Paul Blatchford
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Dr Paul Blatchford, Scientist at Plant & Food Research, New Zealand, presentation at the 1st International Symposium on Kiwifruit and Health: http://www.kiwifruitsymposium.org/presentations/kiwifruit-driven-microbiota-metabolites-and-implications-for-human-health/ The research presented attempts to understand how kiwifruit impacts upon microbial composition and metabolism in the human large bowel using in vitro fermentation systems.
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Kiwifruit-driven Microbiota, Metabolites And Implications for Human Health by Dr Paul Blatchford
- 1. Kiwifruit-driven Microbiota, Metabolites and Implications For Human Health Dr Paul Blatchford Plant & Food Research, New Zealand
- 2. KIWIFRUIT-DRIVEN MICROBIOTA, METABOLITES AND IMPLICATIONS FOR HUMAN HEALTH Paul Blatchford
- 3. Health benefits Glycaemic impact Vitamin C Prebiotic effects? Laxation Immune benefits
- 4. Health benefits Glycaemic impact Vitamin C Prebiotic effects?Laxation Immune benefits Prebiotic definition ‘a selectively fermented ingredient that results in specific changes in the composition and/or activity of the gastrointestinal microbiota, thus conferring benefit(s) upon host health’ (Gibson et al., 2010)
- 5. KIWIFRUIT • Kiwifruit contain 2-3% non-starch polysaccharides (NSP) such as pectic polysaccharides, hemicellulose and cellulose • The NSP escape digestion by human enzymes and reach the colon relatively unmodified. Here they become substrates for bacterial fermentation The following work used green-fleshed Actinidia chinensis var. deliciosa `Hayward’ (Zespri® Green Kiwifruit) and gold-fleshed Actinidia chinensis var. chinensis `Zesy002’ (Zespri® SunGold Kiwifruit)
- 6. GUT MICROBIOTA • The human gut microbiota has strong associations with many aspects of human health • The colon is an active site of fermentation: - short chain fatty acids (SCFA) - branched chain fatty acids (BCFA) - gases (hydrogen, carbon dioxide and methane) • Acetate, propionate, butyrate as well as lactate, formate, succinate etc..
- 7. HOW DOES KIWIFRUIT AFFECT BACTERIAL COMPOSITION?
- 8. • Mimics the large bowel microbial community • Several variables can be controlled: - pH - temperature - O2 • 10% w/v faecal slurry • Typically run in triplicate with different faecal donors • Run for 24 – 48 hours IN VITRO PH-CONTROLLED BATCH CULTURE GUT MODEL
- 9. IN VITRO FERMENTATION STUDY 1 • Batch culture gut models (in vitro) conducted with green and gold kiwifruit - 16S rRNA gene sequencing, Gas Chromatography, NMR Spectroscopy Kiwifruit fermentation drives positive gut microbial and metabolic changes irrespective of initial microbiota composition. P Blatchford, H Stoklosinski, G Walton, J Swann, G Gibson, R Gearry, J Ansell, Bioactive Carbohydrates and Dietary Fibre, 2015.
- 11. IN VITRO FERMENTATION EXPERIMENTS
- 12. ORGANIC ACIDS BY GAS CHROMATOGRAPHY µmol/mL µmol/mL µmol/mL 0 5 10 24 48 Time (h) 0 5 10 24 48 Time (h) 0 5 10 24 48 Time (h)
- 13. IN VITRO FERMENTATION STUDY 2 Batch culture gut models (in vitro) with gold kiwifruit - 16S rRNA gene sequencing, Gas Chromatography, LC-MS In vitro characterisation of the fermentation profile and prebiotic capacity of gold-fleshed kiwifruit P Blatchford, K Bentley-Hewitt, H Stoklosinski, T McGhie, R Gearry, G Gibson, J Ansell, Beneficial Microbes, 2015.
- 14. HEATPLOT OF THE TOP 22 BACTERIAL GENERA
- 15. BETA DIVERSITY UNWEIGHTED BIPLOT Donor 1 Donor 3 Donor 2
- 16. -2.00 8.00 18.00 28.00 38.00 48.00 58.00 Gold - Acetate Gold - Propionate Gold - Butyrate Negative - Acetate Negative - Propionate Negative - Butyrate µmol/mL Time (h) 0 5 10 24 48 After 48 hours, the ratio of the three primary short chain fatty acids was 67:26:7 for acetate, propionate and butyrate, respectively ORGANIC ACID CONCENTRATIONS
- 17. RELATIVE AMOUNTS OF METABOLITES IN THE ZESY002 VESSELS 0 0.1 0.2 0.3 0.4 0.5 0.6 0 5 10 15 20 25 30 35 40 45 50 NormalisedIntensity Time (h) Quinic acid - 191.0562 m/z Citric acid - 191.0211 m/z Malic acid - 133.0143 m/z Tryptophan - 203.0818 m/z E-caffeoyl-3-glucoside - 341.0877 m/z
- 18. • Despite notable donor inoculum differences, kiwifruit was able to drive the microbial profile towards more commensal genera • Kiwifruit supplementation in vitro promoted the commensal Bacteroides spp., Parabacteroides spp. and Bifidobacterium spp. • The kiwifruit-driven increase of commensal bacteria in the in vitro models resulted in a propionate- enriched environment • Organic acids and polyphenols can survive SGD and may act as substrates for bacterial fermentation CONCLUSIONS
- 19. • What fractions of kiwifruit are responsible for causing the microbial and metabolic effects presented in this work? • What is the effect of other kiwifruit-derived metabolites that escape simulated gastrointestinal digestion and reach the colon? We know polysaccharides are principal drivers of microbiota composition • Organic acids - Quinic acid, malic acid, citric acid • Polyphenols – E-caffeoyl-3-glucoside, E-caffeoyl-4-glucoside etc... • Fatty acids – Linoleic acid, linolenic acid, oleic acid UNANSWERED QUESTIONS…
- 20. Laxative effects Oxalate/raphides? Organic acids? Bacteria? Fibre? Actinidin? PARTING MESSAGE….
- 21. Unanswered questions.... ARE THE LAXATIVE EFFECTS OF KIWIFRUIT PARTLY MEDIATED BY BACTERIA?
- 22. Thanks! • To Riddet and Zespri for the opportunity to present at this forum • To everyone in the Food, Nutrition and Health group at PFR, Palmerston North • Supervisors – Juliet Ansell, Richard Gearry, Glenn Gibson QUESTIONS? THANK YOU Paul Blatchford Paul.Blatchford@plantandfood.co.nz 064 6 3556164