Marine bio-actives and health: Functional Foods -...
Transcript of Marine bio-actives and health: Functional Foods -...
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ulster.ac.uk Dr. Chris GillPromac Conference Alesund, Norway, 9th November 2018
Marine bio-actives and health:
Functional Foods
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55.1326° N, 6.6646° W
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Overview
• Global Seaweed Industry
• Functional foods
• Examples of functional food validation
– Seaweed polysaccharides
– Seaweed (poly)phenols
• Summary
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Seaweed production
Aquaculture
Fisheries
Buschmann et al. EJOP. 2017
Wet metric tonnes
Chile, China & Norway lead exploitation of wild stocks of seaweeds.
China & Indonesia largest seaweed producers 23+ MT (aggregate)
Norway – seaweed exploitation largely Laminaria hyperborea , Ascophyllum nodosum)
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Global seaweed industry
• 28.5 million wet tonnes of seaweed harvested (cultivated & wild, 2014)
• In 2015, the global “commercial” market US$ 10.31 billion
• US$ 22.13 billion by 2024, growing at a CAGR of 8.9%.
• In 2012 9 million eaten directly as sea vegetables
• Edible seaweed market in Europe typically growing at 7-10% p.a.
• Human consumption - edible food products, cosmetics, health & wellness = 81% of market value in 2015 - highest growth sector 2016-2024.
FAO report 2014, Grandview Research Report (2016), FAO report 2012 , Hotchkiss et al (2016)
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Typical use in food
Seaweed ingredients added for number of reasons:
• texture – as hydrocolloids (extracts) and whole ingredients;
• nutrition/supplementation – e.g. minerals, vitamins, antioxidants;
• flavour – including sodium replacement;
• interest/novelty.
Seisun et al, IMR, 2016.
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Food hydrocolloids
• Primary use of seaweed derived food ingredients = hydrocolloids.
• In Europe = “additives, E Nos” not “ingredients”.
• The seaweed derived hydrocolloids – carrageenan, agar and alginate – represent 3/5 top food hydrocolloids market sectors in market value (excluding gelatin & starch)
• Estimated global production (exc. China) in 2015 (tonnes dw):
• carrageenan - 68,000;
• agar – 12,500;
• alginate – 16,000.
Seisun et al. IMR, 2016, Sanchez et al 2016
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Seaweed products value pyramid
• < € 1/kg - dried & milled/flaked only
• Seaweed meals for feed and horticulture
• Food grade powders in bulk
• € 1-5/kg - more processing involved
• Specialist feed & horticulture products
• Higher quality food grade powders
• €5-1000/kg – depending on purity & use
• Hydocolloids/bioactive extracts
• >€2000/kg High purity bioactive extracts (polyphenols/polysaccharides) for nutra/cosmaceutical/research.
Desire to increase the value of seaweed & seaweed extracts- functional food?
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A food material that provides specific functional benefit (health or physiological) additional to its
basic nutritional value
A functional food
We’re getting older…
Increased healthcare costs (>8% GDP)
Opportunities for seaweed/seaweed extracts & functional foods?
Sales of functional foods worldwide
from 2015-2024
Statista 2018
Sale
s in
bill
ion
UD
$
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Seaweed & Health
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Seaweed based functional food studies
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HYFFIHydrocolloid derivatives as functional
food ingredients
European Commission (FP7) Grant agreement No 222470
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Seaweed as a source of fibre
• Increased fibre consumption associated with gut health
• 8g portion of seaweed ~ 11% Fibre GDA
• Seaweed is a sustainable source of a range of polysaccharides –
Alginate, Agar, Carrageenan, Fucoidan, Ulvan
• Too complex and relatively inaccessible for human microbiota
• Hydrolysis to form novel LMW derivatives to allow colonic fermentation – prebiotic?
• Applications in food & health, and wellness products
McArtain et al. Nut Rev 2007, Brown et al Nut Rev 2014, Shang et al Carbo poly 2018
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Prebiotic potential
• A non-digestible food ingredient beneficially affects the host, selectively stimulating the growth and/ or activity of one or a limited number of bacteria (e.g. LAB) in the colon that have the potential to improve host health
• Modulation of the human gut microbiotaincrease in Bifidobacterium spp.+/- increase in lactobacilli+/- decrease in harmful bacteriaProduction of SCFA’s
Well known food industry examples inulin,derived from chicory root.
Potential prebiotics effects seaweed extracts, oligo/polysaccharides- mainly in vitro + animal studies
Gibson et al , 2004, Raposo et al, Mar Drug 2016; Rowland et al EJON 2017
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HYFFI Aim - To develop, from agar & alginate polymers, novel prebiotics.
Seaweeds Tested
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Overview
Alginates and agar are dense hydrocolloid compounds that form gels.
Depolymerisation ~ reducing its gelling properties and increasing fermentability & consumer acceptability
19 Candidate LMWPs produced for in vitro screening
NovelLow molecular weight
PolysaccharidesLMWPS
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LMWPs Prebiotic Screening
24h batch culture (37ºC),pH controlled (6.8),
10% faecal slurry, donors (n=3), 1% LMWP.
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Significantly increased Bifidobacteria.
Extensively fermented, significant increase in Total SCFA
Prebiotic index comparable - inulin
Extensively fermented, significant
increase in Total SCFA
Gelidium seaweed extract CC2253Agar candidate
Ascophyllum nodosum extract CC2239Alginate candidate
2 best candidates based on in vitro screening.
Selection of prebiotics for production scale-up
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Scale up issues
Undegraded MW 130,000Degraded MW 4038090% <50,000.Within EU spec
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Scale up issues
Undegraded MW 179000Degraded MW 1173997% <50,000.Within EU spec
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HYFFI Prebiotic efficacy study
• Double blind randomised placebo controlled 3-way crossover trial
• 8g LMW Agar, LMW Alginate, maltodextrin (control). Water based Drink, 3.2% fibre, 6% sugar, Na benzoate, Na citrate, citric acid, flavour & colour.
• N= 60 (30m/30f), BMI >20 <35, 18-55 years
• Duration 21 weeks. No antibiotic or supplements use.
• 6x stool samples, 6x blood samples, 6x 4 d food diary, bowel habits
Treatment28 days
A
Treatment28 days
B
Treatment28 days
C
Tp1 Tp2 Tp3 Tp4 Tp5 Tp6
Washout 28 days
Washout 28 days
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HYFFI Prebiotic efficacy study- Outcomes
Allsopp et al EJON in prep
Increased dietary fibre consumption, stool bulking effects.LMW alginate reduced post prandial glucose, known effect for intact alginate.
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Gap between laboratory and industry
• Industrial scale up – Feasibility and practical issues
• What is your target medium - Product incorporation
• Consumer acceptability – can taste and texture issues be addressed
• Gelidium seaweed, Ascophyllum – never make it to human study?
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PREMARA (Prebiotics from the sea)
Refinement of HYFFI, PreMara in progressFIRM:Premara 13 F 511
Palmaria palmata, Fucus vesiculosus, Laminaria digitate, Ulva spp.
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PREMARA
Crude and depolymerised extracts have significantly increased Bifidobacteria
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Gap between laboratory & industry - Premara
• Industrial scale up – Feasibility and practical issues
• Consumer acceptability –taste and texture issues addressed
PremaraHuman study
Study recruited
(n=60)
Sampling concludes Dec 2018
Analysis starts Jan
2019
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SWAFAX- Seaweed derived anti-inflammatory agents and antioxidants
European Commission (FP7) Grant agreement No 26259
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Gómez-Guzmán et al Marine Drugs 2018
Brown algaePhlorotannins
6 subclasses:eckols,fuhalols, fucophlorethols, phlorethols, fucols, ishofuhalols
Seaweed (poly)phenols
Green, red algaebromophenols, phenolic acidsflavonoids.
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WHO 2014, Rangel-Huerta et al 2015, Del Rio et al 2013, Brown et al 2014, Fernando et al 2016, Gómez-Guzmán et al 2018
SWAFAX- Seaweed derived anti-inflammatory agents and antioxidants
• Cardiovascular disease (CVD) is the leading cause of death worldwide
• Evidence suggests a positive effect of (poly)phenol intake on CVD risk
• (Poly)phenols have antioxidant and anti-inflammatory properties
• Brown seaweed is a rich source of (poly)phenols
• No information on the bioavailability of seaweed (poly)phenols, a critical factor influencing in vivo biological activity.
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Preparation of food-grade seaweed polyphenol extract and capsule
Fresh A. nodosum was supplied by The Hebridean Seaweed Company, Isle of Lewis, Scotland in March 2011
Seaweed extract Ascophyllum Nodosum (SPE)
Solvent extracted 60:40 ethanol–waterA solvent– seaweed ratio of 3:1
Characterised (NP)-HPLC, LC-MS analysis
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24 healthy volunteers (12 M, 12F), age 18 – 65 years.
Acute bioavailability study
SPE capsule - 400mg Capsule, 100mg polyphenols ~Phlorotannins - Hydroxytrifuhalol A, 7-Hydroxyeckol, C-O-C dimer phloroglucinol
de-glucosilation
methylation
glucuronidation
Gut-bacteria
Fermentation
Small phenolics
Cannulation, blood & urine samples collected.
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SPE Metabolites in Urine
Corona et al BJN 2016
LC/MSN - urine post seaweed consumption.
Hydroxytrifuhalol A7-Hydroxyeckol C-O-C dimer of phloroglucinol
HPLC-DAD – 18 conjugated(glucuronides/sulfates)un-conjugate seaweed metabolitesdetected– Phoroglucinol standard
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Seaweed Metabolites in Plasma
Conjugated (glucuronides/sulfates)un-conjugate seaweed metabolites detected.
Corona et al BJN 2016
HPLC DAD- Seaweed metabolites found in samples collected at 0-8h after capsule ingestion, but majorityof the metabolites found in samples collected at 8-24h.
Anti- inflammatory activity Ex-vivo LPS + Whole blood culture–increase cytokine IL-8
IL-8
Importance of Colonic Fermentation HMW PPs?
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Antioxidant activity decreases with digestion,Fermentation.
CF
GID
ND
Colonic fermentation increasedAnti-genotoxicity (HT29 cells)
CF HMW most potent
Colonic Fermentation HMW PPs
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N= 80, 30-65 yrs, BMI>25
Registry no. NCT02295878.
SWAFAX study
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Am J Clin Nutr 2018;108:688–700.
Untargeted analysis of urine and plasma-UHPLC-HR-MS
Biomarkers of seaweed consumption identified in urine only
Total seaweed metabolites excreted0.001 - 4.140 mmol.
25% low excreters<0.5mmol
20% low excreters<2.0mmol
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Seaweed extract significantly reduced basal DNA damage in PBMCs,in population at highest risk (obese).
N=78 N=42 N=36
individual treatment effects (pre – post supplementation), * P>0.05, (ANOVA, one-tailed test)
Average basal DNA damage 6.91 +/- 1.9
23%
SPE & Risk Biomarker
Peripheral Blood Mononuclear Cells (PBMCs)
Baldrick et al; AJCN 2018
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SWAFAX study key outcomes
A range of metabolites (un-conjugated & conjugated forms), especially at the later time points. (6 to 24 hours) - due to colonic metabolism of high MW phlorotannins?
High & low seaweed phenolic excretors were observed.
Decreased lymphocyte DNA damage in obese individuals.
Consumption of seaweed phenolics for 8 weeks had no negative effects on:
C-Reactive Protein, Plasma total cholesterol, HDLC, LDLC or triglycerides levels.
IL-1, IL-2, IL-6, IL-10, IL-12, IFN- and TNF- expression in mononuclear cells
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Double blind, randomized controlled, parallel study, n=40, 18-65 y, 5 g dulse p.d.in 4 weeks.
P. palmata extracts tested in vitro first, was anti-inflammatory.P. palmata bread stimulated inflammation (CRP), elevated serum triglycerides, altered thyroid function; changes within clinically normal range not likely to impact health.
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Seaweed extract no effect on plasma glucose or plasma insulin response
Randomised Double Blind, Placebo Controlled Crossover Trial: N=38; aged 19-56 y Fucus vesiculosus extract 500 mg, 2000 mg , Placebo cellulose - 2000 mg
plasma insulin responseplasma glucose
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Computerized measures of episodic memory, attention/subjective state.
500 mg A. nodosum & F. vesiculosus
Randomised Double Blind, Placebo Controlled Parallel Trial: N=60; aged 18-65 yrs.
Seaweed improved accuracy on digit vigilance (p = 0.035) & choice reaction time (p = 0.043)
Hot water extract (Sea2®)
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Summary
• Global “commercial” market for seaweed US$ 10.31 bn
• Edible food products/cosmetics/health & wellness and pharma for
human consumption = the highest market share.
• Majority of seaweed and health studies in vitro /animal
• Functional food requires validation in humans
• Seaweed fibre (LMWP agar) effects gut function and microbiota
• Seaweed (poly)phenols are bioavailable & bioactive in humans
• Opportunities for seaweed/seaweed extracts as functional foods
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• Professor Sean Strain Dr Philip Allsopp
• Professor Ian Rowland Dr Kirsty Pourshahidi
• Dr Emeir McSorley Dr Emma Brown
• Dr Pamela Magee Dr Mary Slevin
• Dr Julie Wallace Ms Vicky Hegan
• Dr Sonja Nitecki Mr Ken Collins
• Dr Thomas Smyth Dr Anna Soler
• Dr Catherine Stanton NutraMara colleagues
• Mr Chris McLaughlin Dr Mary Rae
• Prof Paul Ross Dr Sarah Hotchkiss
• Dr Gema Periro-Caro Mr Ross Campbell
Acknowledgments
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Any questions?