Benefits of Prebiotic Oligosaccharides as Ingredients for Specific Milk Formulations for Young Children
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1 Benefits of Prebiotic Oligosaccharides as Ingredients for Specific Milk Formulations for Young Children Jacques G. Bindels, PhD Danone Research Danone Research Centre for Specialised Nutrition Amsterdam / Wageningen, The Netherlands
2 agenda agenda Prebiotics and prebiotic oligosaccharides sources and structures t minimally effective dosage Specificity of beneficial effects of prebiotic oligosaccharides Beneficial effects on the immune system
3 prebiotics Prebiotics must survive acidic conditions Prebiotics must evade digestion in the small intestine Prebiotics must stimulate one or a limited number of beneficial Prebiotics must be bacteria, like bifidobacteria or selectively l fermented in lactobacilli, within the colonic the colon microbiota Gibson & Roberfroid (2004) Nutr Res Rev 17:
4
5 Inulin & oligofructose/fructo-oligoaccharides Inulin is industrially isolated from chicory roots and shows a large size heterogeneity with a DP from 2 to approx. 60 In chicory inulin approx.: 1/3 short chain: DP 2-9 2/3 long chain: DP >10 Short chain inulin = FOS - made out of inulin -made chemically Long chain inulin = lcfos= polyfructose - made out of inulin [Frc(β2-] )Frc(β2-1)Glc
6 Galacto-oligosaccharidesoligosaccharides Derived from lactose by enzymatic trans-glycosylation Originally called TOS, now more often GOS [Gal(β1-4)] 0-5 Gal(β1-4)Glc GOS has a narrow size heterogeneity: from DP2 to DP6, fairly comparable to (short chain) FOS
7 Research on prebiotics and gut health in infants and young children in infants and young children Studies with single type prebiotics GOS FOS ~ oligofructose Inulin Native inulin Polyfructose (long chain FOS) Studies with mixtures of prebiotics GOS + polyfructose (long chain FOS) GOS + (short chain) FOS GOS + polydextrose + lactulose
8 Studies with GOS only Ben et al. 2004, 2008 (Nanjing, China) age 1-6 mo. amount 0.24 g / 100 ml - per day approx g stimulation Bifido s & Lactobacilli, stool charateristics Fanaro et al (Ferrara, Italy) age 4-8 mo. amount 0.5 g / 100 ml - per day approx g stimulation Bifido s, stool characteristics
9 Studies with FOS only Brunser et al 2003, (Santiago, Chile) age 3-6 mo amount 0.2 g/100ml - daily: approx. 2 g stimulation bifido s only relative little effect Euler et al 2005 (Norfolk, USA) age 1-3 mo amount 0.15 & 0.3 g/100ml daily: 1-2 g no stimulation bifido s more loose & frequent stools Waligora-Dupriet 2006 (Paris, France) age 7-19 mo adminstered as supplement - daily: 2 g bifido s tended to increase, good tolerance less flatulence
10 Studies with inulin only Yap et al 2005 (Selangor, Malaysia) age 5-12 months administered as supplement: daily 0.75, 1, 1.25 g no effect on SCF, but effect on Fe and Mg app retention Yap et al 2008 (Selangor, Malaysia) age 5-12 months administered as supplement: daily 0.75, 1, 1.25 g at highest dosage: significant increase bifido s, decline coli s, clostridia and gram+ cocci, sign decrease in stool ph Kim et al (Seoul, Korea) age 3-5 months 0 25 g/kg/day daily approx 1 4 g 0.25 g/kg/day daily approx. 1.4 g stimulation of bifido s, tendency for stool characteristics
11
12 Conclusions so far Out of the portfolio of clinical i l studies with infants and babies, bi it may be concluded that the minimal effective dosage for GOS and Inulin (natural mixture of 1/3 FOS and 2/3 polyfructose) is about 1.5 g / day Based on a total daily intake of at least 750 ml, the minimal effective concentration is 0.2 g / 100 ml (short chain) FOS alone is probably somewhat less effective as a prebiotic and needs a minimal effective dosage of about 2 g / day Based on a total daily intake of at least 750 ml, the minimal effective concentration is 0.3 g / 100 ml The difference in efficacy between (short chain) FOS and inulin is The difference in efficacy between (short chain) FOS and inulin is supported by published in vitro research.
13 GOS / polyfructose mixture 9:1 Inspired by human milk oligosaccharide size distribution Studied in 16 independent clinical trials with 2670 babies data from wide age range: preterms, neonates, infants, toddlers positive results reported at intake levels from 2.3 to 6 g / day unpublished data: limited results at intake level of 1.15 g / day support minimal effective amount of 1.5 g /day also for GOS/polyfructose Beneficial effects gut health parameters microbiota (= prebiotic effect!), fecal SCFA, lactate and ph stool consistency (very consistent) and frequency (in preterms) immune system related parameters
14 Immune system related effects of GOS/polyfructose prebiotic oligosaccharides (1) Stimulation of secretory IgA production by gut immune cells (siga levels in the feces at age 6 months. Scholtens et al., 2008) Faecal sig ga (µg/g fece es) * GOS/polyfructose Control Human milk *) Significant difference between GOS / polyfructose and control p<0.05 Unclear whether this effect is independent of the microbiota as it is also seen in some studies with probiotic supplementation
15 Immune system related effects of GOS/polyfructose prebiotic oligosaccharides (2) Reduction in infections in 2 independent studies URTI (2 studies), diarrhea (1 study), antitbiotic use (2 studies) Persistant effect at age 2 years (1 study) despite that dietary intervention was only until age 6 months Reduction in atopic dermatitis Study in children at high familiy risk to develop allergy using HA formula Persistant effect at age 2 yr despite that dietary intervention was only until age 6 mo First sign for effects on rec.wheezing placebo GOS/polyfructose
16 Specificity within effects prebiotic oligosaccharides For the effects on intestinal ti micobiota per se: NOT (inherent to definition!) For the minimally effective dosage: YES For tolerance / side effect aspects: (= maximally tolerated dosage) YES adults: GOS tolerated without side effects at higher dosages than FOS babies: reported minor tolereance problems together with unclear bifidogenic effects with 0.3 g/100 ml FOS: EFSA rejection of application For effects exceeding the prebiotic concept: (so: effects not mediated via the intestinal microbiota) YES
17 Influenza vaccination model in C57BL/6 mice Animals were prefed with prebiotics during 2 weeks prior to the first of 2 vaccinations 2 prebiotic mixtures depicted: both groups showed similar prebiotic activity in only 1 group: Th1 immune enhancement Vos et al. (2006) Int Immunopharmacol 6:
18 The specific GOS-polyfructose mixture directly affects the adaptive immune system Th1/Th2 balance via DC/T-cell interaction stimulation of anti-allergic profile Ratio IFN-γ γ / IL-4 (positive cells) 6,0 5,0 ** IFN-γ: Th1 marker IL-4: Th2 marker 4,0 3,0 2,0 1,0 Control GOS/polyfructose 0,0 Garssen et al., 2010 (in press) 18
19 Direct effect of polyfructose of IFNγ induced stimulation on NO+NO 2 production by human innate immune system macrophage/monocyte RAW264.7 cells 60 Raftiline HP = polyfructose - NO 2 productio on (um) Raftilose P95 = (short-chain) FOS 0 Medium IFN-γ Rafiline/Raftilose (mg/ml) + IFN-γ Garssen et al., 2010 (in press)
20 Pectin derived Acidic OligoSaccharides paos: active Compound of Carrot Soup 11th Century Health effects of carrots (esp. for gut problems) known for nearly 1000 years (Macer Floridus: De Viribus Herbarum, 11th century) 1908 Carrot Soup as Treatment for Diarrhoea (Moro E, Münchn Med Wschr 1908; 31: ) 1997 Acidic Oligosaccharides (Oligogalacturonic acid) identified as anti-adhesive compounds (Guggenbichler JP et al. Pharm Pharmacol Lett ; 1:35-38) % of Human milk Oligosaccharides have a negative charge (acidic structures) and may be function as anti-adhesive compounds (Stahl B,et al, Anal. Biochem. 1994, 223, 218)
21 Chemical Structure of paos
22 Animal Studies: Both GOS/polyfructose and paos improve vaccination responsiveness (~non-virulent infection) The combination gives synergy Vos AP et al.: Acidic oligosaccharides enhance systemic immune responses in mice: dose response, interaction and comparative experiments in a murine vaccination model; J.Pediatr. Gastr. Nutr. 2005, 40:616 Delta ear thic ckness (μm + sd d) DTH response (~cellular immunity) * * * #
23 Multi-Centre Infection & Eczema Prevention Study 7centrersin5EUco countries 1130 babies enrolled 2 formula groups + paos/gos/polyfructose Without prebiotics (control) Breast feeding group (reference) Intervention started before age of 8 weeks and continued until age 12 months Babies with high risk to develop atopy (family history) were not included Main outcome parameter febrile infections did not reach stat significance due to very low number of infecions Gruber et al., 2010 JACI (e-pub ahead of print)
24 Conclusions The minimally effective dosage of prebiotic oligosaccharides in young children (0-1 yr) is about 1.5 g / day for GOS, inulin and polyfructose and probably somewhat higher for FOS Next to effects via stimulating the intestinal microbiota, some prebiotic oligosaccharides have additionally direct effects/interactions with the immune system Clinically relevant beneficial effects on the immune system have been reported only for GOS / polyfructose mixture a decreased risk for infection a decreased risk for atopic dermatitis and possibly recurrent wheezing beneficial effects may be persistent even after ending dietary intervention paos is promising as an additional ingredient to synergistically improve the functionality of the GOS/polyfructose mixture
25 THANK YOU terima kasih
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