http://apc.ucc.ie http://apc.ucc.ie

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DEPARTMENT OF

UNIVERSITY COLLEGE CORK

Gut Microbiota, Diet and Health in

the Elderly Population

Marcus Claesson

1st

October 2012

ISAPP

“Gut microbiota as an indicator and agent for nutritional health in elderly Irish subjects”

Why elderly? • Increasing proportion in population

• Changes in microbiota composition and activity

• Increased infection rates

• Increased inflammatory diseases

• Prospects for dietary intervention/modulation

How does the gut microbiota

composition change with age?

Infancy

• Simple

• Temporal instability

• Dominated by Bifidobacteria

Adulthood

• More complex

• Temporal stability

• Dominated by Firmicutes & Bacteroidetes

Physiological changes of the aging intestine:

• Reduced motility => altered nutrition dynamics

• Reduced dentition & taste => altered diet

• “Inflamma-aging” => chronic low-level inflammation

How does the gut microbiota

composition change with age?

•Elderly gut microbiota in a state of flux (Mitsuoka et al. 1978)

•Conflicting age-related changes of the major

phylogenetic groups:

• Bacteroidetes up; Actinobacteria down (Hopkins et al. 2001)

• Bacteroidetes up; Firmicutes down in Irish (Claesson et al. 2011)

• No Bacteroidetes change for 100yr old Italians compared to 30

& 70yr olds (Biagi et al. 2010)

• Composition does not vary between European countries (Lay et al. 2005)

• Composition does varies between European countries

(Mueller et al. 2006)

Is community location of the elderly

associated with microbiota?

• Location: proxy for general health condition

• 178 elderly (≥65yrs) Irish subjects • 83 Community-dwelling

• 20 Day hospital (out-patient)

• 15 Rehabilitation (≤6 weeks)

• 60 Long-stay (>6 weeks)

• (13 Young healthy controls)

• No antibiotics treatment ≤1 month prior sampling

• Collected stools samples • 16S rDNA amplicons (454) & shotgun (Illumina) sequencing

• Metabolomics (NMR)

• Food Frequency Questionnaire => long-term diet

• BMI, frailty, malnourishment, depression, cognitive

function & dementia

5.4mio 16S rDNA reads => 47,500 OTUs

Subjects separated by community location

Community Long-stay Young control

Unweighted UniFrac OTU PCoA Weighted UniFrac OTU PCoA

Subjects separated by community location

Hierarchical Ward-linkage clustering based on Spearman

correlation coefficients of the proportion of OTUs for each subject

Unweighted UniFrac OTU PCoA

Enriched in Community (p

What impact has diet on microbiota?

Food Frequency Questionnaire (FFQ)

• Long-term dietary habits

• FFQ data for 96% elderly subjects

• 147 food types (beef/apples/white

rice/potatoes/milk/porridge etc)

• Healthy Food Diversity (HFD): how

diverse AND healthy a diet is

FFQ multivariate analysis

Correspondence analysis

FF

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ypes

Complete-linkage clustering based on

Euclidean distances to PC1

DG1: “low fat / high fibre”

DG2: “moderate fat / high fibre”

DG3: “moderate fat / low fibre”

DG4: “high fat / moderate fibre”

Diversity of microbiota and diet

Microbiota & diet

by community location

Unweighted UniFrac PCoA vs. FFQ PCA Weighted UniFrac PCoA vs. FFQ PCA

Diet Microbiota

Community Long-stay

Microbiota & diet

by duration in long-stay care

N/A (C+DH)

Diet Microbiota

Week0to6 (Rehab) Week6toYear1 Year1+

Enterotypes

• Arumugam et al., 2011 Nature:

• 39 individuals from 6 countries (+239 US/DK individuals)

• “All people can be classified into 3 enterotypes”

• Dominant genera:

• Bacteroides

• Prevotella

• Ruminococcus (Blautia / Lachnospiraceae)

• Wu et al., 2011 Science:

• 98 individuals from the US

• Only Bacteroides & Prevotella enterotypes stable

• Stable over time

• Associated with long-term diet

• Bacteroides: high-fat/low-fibre

• Prevotella: low-fat/high-fibre

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Enterotype clustering in the elderly

• 29 subjects representative of C/R/LS

• Metabolomics (n = 29)

– NMR spectroscopy of faecal water

– Spectra -> bins -> metabolites (PCA)

• Shotgun metagenomics (27 of 29)

– Total extracted bacterial DNA sequenced

– 51mio 2x91bp Illumina reads/sample

– 126 Gb of DNA sequenced

– 2.51mio predicted genes

Microbiota function:

Metabolomics and Metagenomics

Pairwise PLS-DA of NMR spectra

Community Rehab Long-stay

Dr. Martina Wallace and Dr. Lorraine Brennan, Univ. College Dublin

Separation of location by faecal

water metabolome

Co-inertia of

microbiota &

metabolome

coloured by

location

NMR spectrum

metabolite PCA

Integrating metabolome & microbiota

Associated microbiota at genus level

Shotgun metagenome:

differentially abundant SCFA genes

BCoAt: Butyryl-CoA transferase / Acetyl-CoA hydrolase

ACS: Acetate-formyltetrahydrofolate synthetase / Formate-tetrahydrofolate ligase

PCoAt: Propionyl-CoA:succinate-CoA transferase / Propionate CoA-transferase

Butyrate Propionate Acetate

Inflammatory markers vary by

community location

Health/clinical markers

• BMI: Body Mass Index

• CC: Calf Circumference

• MAC: Mid-Arm Circumference

• SBP: Systolic Blood Pressure

• DBP: Diastolic Blood Pressure

• CCI: Charlson Index of Comorbidity

• Barthel Index of Activities of Daily Living

• FIM: Functional Independence Measure

• MMSE: Mini-Mental State Exam

• MNA: Mini-Nutritional Assessment

Microbiota-health correlations

Possible confounders

– Antibiotics:

• Exclude 1mo had no sign. effect on

µ-biota (α- or ß-diversity)

– Quantile regression

model adjusted for:

•Age

•Gender

•Location

•Medication

Microbiota separation

correlates with health measures

Location-specific unweighted UniFrac PCoAs

Community-only subjects

All four location subjects

Long-stay-only subjects Community Long-stay

Following adjustment for age/gender/location/medication, microbiota

correlates significantly with e.g. frailty and inflammation.

Prospective studies needed to establish causality.

How to reduce complexity of

microbiota composition?

Co-abundance groups (CAGs): groups of genera that are

positively correlated with each other

Microbiota changes across location

is mirrored by changes in health

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Summary

(Claesson, Jeffery et al., 2012 Nature)

• Microbiota in elderly is different depending

on community location

• Driven by habitual diet

• Microbiota alterations correlate with health

changes especially in long-stay

Diet shapes gut microbiota, which might

impact on health in elderly people

May lead to carefully designed dietary supplements

to promote healthier aging

Acknowledgements

Paul O’Toole

Anthony Fitzgerald

Denis O’Mahony

Paul Ross

Catherine Stanton

Gerald Fitzgerald

Fergus Shanahan

Ted Dinan

Martina Wallace

Julian Marchesi

Lorraine Brennan

Michael O’Connor

Douwe van Sinderen

Colin Hill

Cillian Twomey

Kieran O’Connor

Lorraine Brennan

Ian Jeffery

Eibhlis O’Connor

Siobhán Cusack

Hugh Harris

Susana Conde

Jennifer Deane

Orla O’Sullivan

Mary Rea

Colm Henry

Mairead Coakley

Patricia Egan

Susan Power

Karen O’Donovan

Ann O’Neill

Norma Harnedy

Bhuna Laks

Martina Wallace

The Cork City

Geriatricians Group