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SCIENTIFIC PUBLICATIONS FACT SHEETS

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NOTE

The strain Biﬁ dobacterium animalis sp. lactis DN-173 010 is deposited in the Collection Nationale de Cultures de Micro-Organismes (CNCM), at the Institute Pasteur (Paris, France) as Biﬁ dobacterium animalis sp. lactis CNCM I-2494.

The nomenclature DN-173 010 can be found in several scientifi c publications...... PDF ACTIVIA.indd 2 15/11/12 13:51 PDF ACTIVIA.indd 3 physiological digestiveevents. to subjects with IBS. IBS subjectshavemoreseveregastrointestinalsymptoms IBS to subjectswithIBS. There isacontinuousincreaseoftheseverityandfrequency ofthesedigestivecomplaintsfromhealthysubjects subjects towithIrritableBowelSyndrome(IBS). Digestive complaintsarewidelyexperiencedbyalargepart ofthegeneralpopulation,rangingfromhealthy bowel habit(constipation,diarrhea),leadingtodiscomfort(cf. fi gure below). It is affected by digestive complaints such as abdominal pain, fl atulence, bloating, borborygmi and altered It ismultifactorial,andincludesphysicalpsychologicalcomponentsthatareintegratedasglobalperception. quality oflife.Thisisthusanimportantaspecthumanhealth. Gastrointestinal comfortreferstotheabilityofdigestivesystemfunctionproperlyandsupportagood INTRODUCTION BORBORYGMI LOOSE STOOLS DIARRHEA 2 DISCOMFORT CONSTIPATION/ HARD STOOLS BLOATING GI 1 , andahighersensitivityto ABDOMINAL PAIN FLATULENCE 15/11/12 13:51

INTRODUCTION PDF ACTIVIA.indd 4 INTRODUCTION Today, atotalof9clinicaltrialsonthemechanismsactionActivia throughout theGItract.Thisparthasbeenenrichedrecentlywithfi rst insightsongutmicrobiotamodulation. movements. OtherstudiesdemonstratetheabilityofstrainBifilactis CNCMI-2494tosurvive dobacterium In addition,studiesevaluatesomemodeofactiontheproduct,suchaseffectsontransittimeandbowel or abdominaldistensionwithphysicalmeasurements. constipation predominant), andevaluate as primary criteria either digestive comfort with questionnaires Four clinicalstudiesconsistentlydemonstratethatActivia parameters (abdominaldistension). as individualdigestivecomplaints(abdominalpain/discomfort,bloating,fl atulence, borborygmi)orphysical allow subjectstoself-report theirdiscomfortlevel. comfort (cf. partActivia Veldhuyzen vanZanten,S.J.O. -Designoftreatmenttrials forfunctionalgastrointestinaldisorders-Gastroenterology2006. syndrome -Aliment.Pharmacol.Ther. 2003.10. Muller-Lissner, S.,Spiller, R.C.,Tack, J. andWhorwell,P.J. -Clinicaltrialguidelinesforpharmacologicaltreatmentofirritablebowel Greece. - Eur. J. Gastroenterol. Hepatol. 2009. G. -Prevalence,bowelhabitsubtypesandmedicalcare-seekingbehaviour ofpatientswithirritablebowelsyndromeinNorthern Zavos,C.,Pilpilidis,I.andChatzimavroudis, Koutras,C.,Gelas,G.,Tziomalos,K., G.,Oikonomidou,I.,Mimidis,K., Paroutoglou, irritable bowelsyndrome-aEuropeanperspectiveAliment.Pharmacol. Ther. 2006.8. Ther. 2004.7.Tack, Spicak,J. J.,andFisher, Fried,M.,Houghton,L.A., G.-Systematicreview:theeffi cacy oftreatmentsfor F.Cremon, C.,Salvioli,B.,DePonti, andCorinaldesi,R.-Diagnosistherapyofirritablebowelsyndrome.Aliment.Pharmacol. W.E. -AGAtechnicalreview onirritablebowelsyndrome.Gastroenterology2002.6. for practiceguidelinedevelopment.–Gastroenterology1997.5.Drossman,D.A., Camilleri,M.,Mayer, andWhitehead, E.A. – Gastroenterology2006.4. 3. Longstreth,G.F., Thompson,W.G., Chey, W.D., Mearin, F. Houghton,L.A., andSpiller, R.C.2006.-Functional boweldisorders Trudgill, N.andWhorwell,P.J. -Guidelinesontheirritablebowelsyndrome:mechanismsandpracticalmanagementGut2007. J. Behav. Med.2002.2.Spiller, R.,Aziz,Q., Creed,F., Hungin,P., Houghton,L.A., Emmanuel,A., Jones,R.,Kumar, D., Rubin,G., 1. REFERENCES: these parameters. bloating, fl atulence) anduponbowelfunction,itismeasuredwithoverallassessmentsthatintegrateall Since gastrointestinalcomfortisdependentupontheperceptionofsensationsarisingfromgut(e.g. conducted onadultshavingminorGIdiscomfort. intervention intendedtoimprovethegastrointestinalcomfortongeneralpopulation,inadditionefficacy trials Scientifi as a sensitive and appropriate model to demonstrate the effic experts recognized IBS cacy of a dietary ischaracterisedbythepresenceofabdominalpainordiscomfort,associatedwithaltered bowel function. IBS digestive comfortbenefi t (cf. partActivia up to20%ofthegeneralpopulationinEuropeandNorthAmerica. This isnotalife-threateningconditionbutitthemostcommonchronicgastrointestinaldisorders,affecting Leibbrand, R.,Cuntz,U. andHiller, W. -Assessmentoffunctionalgastrointestinaldisorders usingtheGastro-Questionnaire.Int. 3,9,10 Theseassessmentsshouldbeperformedwithvalidatedtools,questionnaires,which ® Drossman, D.A., Whitehead,W.E. andCamilleri,M.-Irritablebowelsyndrome:atechnicalreview Effi cacy studies).2studiesareonhealthypopulationandIBS-C(IBS- 9. Irvine, E.J., Witehead, W.E.,Irvine, E.J., Witehead, Chey, W.D., Shaw, Matsueda,K., M.,Talley, N.J. and Corazziari, E., Bytzer,Corazziari, E., P., Delvaux, M., Holtmann, G., Malagelada, J.R., Morris, J., ® Mechanismofactionstudies). 10 Severalotherparameterscanalsobemeasuredsuch ® consumptionimprovesgastrointestinal 4-8 De Giorgio,R.,Barbara,G.,Stanghellini,V., Katsinelos, P.,Katsinelos, Lazaraki,G.,Kountouras, J., ® accompanythescienceon 3

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...... 1. ACTIVIA® EFFICACY STUDIES ......

STUDIES ON HEALTHY POPULATION Guyonnet et al., 2009 a Guyonnet et al., 2009 b

STUDIES ON IBS-C POPULATION Agrawal et al., 2009 Guyonnet et al., 2007 ...... PDF ACTIVIA.indd 5 15/11/12 13:51 PDF ACTIVIA.indd 6 15/11/12 13:51 PDF ACTIVIA.indd 7 or 2x125gservings(n=147)ofActivia self-reported digestivediscomfortandanormalstoolfrequency, consumeddailyeither1x125gserving(n=144) The studywasarandomized,controlled,open-label,parallelgrouptrial. 360healthyvolunteers(18–65yo),with STUDY METHODOLOGY Digestive Diseases,2009;10:61-70. general populationofadults.Arandomized,open-label,controlled,pilotstudy. Journalof Biﬁlactis dobacterium Guyonnet D,Woodcock StefaniB,Trevisan A, C,HallC.Fermented milkcontaining GUYONNET EVALUATION CRITERIA baseline, andattheendofproductconsumption). completed, 20-itemquestionnaire;evaluationat consumption). Likert scale;evaluationafter14daysofproduct digestive comfortchanged?”,usinga5-point “Think backoverthelast2weeks,howhasyour question after14daysofproductconsumption: intervention.

Bother fromdifferentdigestivesymptoms(Self- Self-reported changeindigestivecomfort(one DN-173 010improvedself-reported digestivecomfortamongsta et al.,2009a ® , over14days.Thecontrolgroup(n=69)followedtheirusualdietwithout For bothActivia Digestive symptoms: and 2-potgroups. There wasnosignifi cant differencebetween1-pot control group(p<0.001). higher inbothActivia improvement ofdigestivecomfortwassignifi cantly The percentageofsubjectsreportingan comfort: Self-reported changesingeneraldigestive RESULTS digestive troubles. feeling heavy, lethargicoruncomfortabledueto excessive ortrappedwind,swollenstomachand all symptomscores,includingbloatedfeeling, greater improvement(p<0.001)vs.controlforalmost the twoActivia The between-groupdifferencesconfi rmed eachof 2-pot group84.3%)vs.thecontrol(2.9%). ® groupsdifferedsignifi cantly fromthe ® groups,therewasasignifi cantly ® groups(1-potgroup82.5%; 15/11/12 13:51

ACTIVIA® EFFICACY STUDIES: HEALTHY POPULATION GUYONNET et al., 2009 a

CONCLUSION Daily consumption of 1 or 2x125g of Activia® servings during 14 days may exert positive effects on self- reported digestive comfort in a general population of healthy adults in real-life conditions.

Beyond this global improvement, the bother from most of the digestive symptoms assessed may be reduced in a high and signiﬁ cant percentage of participants.

CHANGE IN PERCEIVED DIGESTIVE COMFORT AFTER ACTIVIA® OR CONTROL CONSUMPTION

100 *p<0.001 vs. control 90 * 80 *

30 Percentatge of participants (%) Percentatge 20

0 WORSE NO CHANGE BETTER

Activia® 1=pot Activia® 2=pot Control

Scores are expressed as percentage of participants per category (worse, no change or better digestive comfort).

PDF ACTIVIA.indd 8 15/11/12 13:51 PDF ACTIVIA.indd 9 The subjectsconsumeddaily2x125gservingsofeitherActivia digestive symptomsbutwithoutgastrointestinaldisorders. The studywasarandomized,double-blind,controlled,parallelclinicaltrialon197women(18–60yo)withminor STUDY METHODOLOGY GUYONNET to theendofstudy): gastrointestinal well-being(weekly, fromweek1 Overall self-reported assessmentof EVALUATION CRITERIA parallel, controlledstudy. BritishJournalofNutrition,2009;102(11):1654-62. symptoms inwomenreportingminordigestivesymptoms:arandomized,double-blind, Bifilactis dobacterium MhamdiL,JakobS,ChassanyO.Guyonnet D,SchlumbergerA, Fermented milkcontaining Bowel function(daily): rumbling stomach. bloating, fl atulence/passage ofgas,borborygmiand digestive symptoms:abdominalpain/discomfort, end ofthestudy): Digestive symptoms(weekly, fromweek1tothe worsening orimprovement. period). consumption) andat8weeks(endofwashout General Well-Being Index). Benefi t Assessment)andPGWBI(Psychological weeks (endofwashoutperiod)): weeks (endofproductconsumption)andat8 Health RelatedQualityofLife(atbaseline,at4 movement/week). was anadditionalfollowup4weeksafterthecessationofproductconsumption(washoutperiod). dairy productwithlowcontentoflactose(lactosesimilartothetestproduct)(n=97),over4weeks.There

5-point Likertscale. Composite scoreandfrequencyofindividual 15-point Likertscaletoprecisethelevelof 3-point Likertscale:worse,nochange,better. Completion atbaseline,4weeks(endofproduct Self-administration of2questionnaires: FBA(Food Stool consistency(BristolScale). Stool frequency(expressedasnumberofbowel DN-173 010improvesgastrointestinalwell-beinganddigestive et al.,2009b groups. control group. Stool frequencydidnotdifferbetween higher intheActivia in theirGIwell-beingwassignifi cantly (p=0.006) The percentageofwomenreportinganimprovement Gastrointestinal well-being DOUBLE-BLIND PERIOD: RESULTS consistency wasobservedintheActivia A signifi cant (p=0.02)improvementofthestool Bowel function abdominal painordiscomfortscores. No signifi cant differenceswereobservedinbloating, group atweeks1,2and4(p<0.05). signifi cantly higherintheActivia The decreaseinfl atulence frequencywas to thecontrolgroup. [95% CI-0.40;-0.04])intheActivia borborygmi frequency(p=0.016;LSmean=-0.22 LS mean=-0.57[95%CI-1.12;-0.02])andin score ofdigestivesymptoms(p=0.044; showed asignifi cant decreaseinthecomposite Over the4weeksofproductconsumption,results Frequency ofdigestivesymptoms [95% CI1.09-3.40]). (p=0.025; 52.0%vs.36.1%respectively, OR=1.92 percentage ofrespondersforGIwell-being (OR=1.69, [95%CI1.17-2.45]),aswellthe ® (n=100), or 2x125g servings of a non fermented (n=100),or2x125gservingsofanonfermented ® groupvs.control ® groupvs.control ® groupcompared ® the groupvs.the 15/11/12 13:51

ACTIVIA® EFFICACY STUDIES: HEALTHY POPULATION GUYONNET et al., 2009 b

Health Related Quality of Life The percentage of responders strongly decreased ® FBA digestive comfort dimension score (primary without signifi cant differences between the Activia and HRQoL endpoint), signifi cantly increased (p=0.027) the control groups (8.0% vs.11.3%; OR 0.68 [95%CI after 4 weeks of product consumption in the Activia® 0.26; 1.77]). group vs. control group (FBA questionnaire). No The composite score of digestive symptoms difference was observed between groups for other signifi cantly decreased (p<0.05) over the 4-weeks in HRQoL dimensions. PGWBI scores did not differ the control group when compared with the Activia® between groups over time. group. No signifi cant difference in the changes of frequency for each individual symptom was observed WASHOUT PERIOD: between the groups. The percentage of women reporting an improvement No differences were observed between groups for in their GI well-being did not differ signifi cantly between stool frequency, consistency or HRQoL dimensions. groups at the end of the washout period.

CONCLUSION The daily consumption of 2x125g servings of Activia® during 4 weeks signiﬁ cantly improved GI well- being and digestive symptoms in the general population of women reporting minor digestive symptoms. The effect of Activia® was no longer observed once consumption was stopped.

OVERAL ASSESSMENT OF GI WELL-BEING (RATE OF RESPONDERS) 60 ® * *p=0.025 Activia 50 Control 40

well-being (%) A responder was deﬁ ned as a subject having an improvement of their gastrointestinal well- 10 Rate of responders for GI being at least 2 weeks among the 4 weeks; 0 results are expressed as percentage. 4 weeks 4 weeks double-blind period washout period

PDF ACTIVIA.indd 10 15/11/12 13:51 PDF ACTIVIA.indd 11 2009; 29(1):104-114. PharmacologyandTherapeutics, irritable bowelsyndromewithconstipation.Alimentary Biﬁlactis dobacterium P.J. JakobS,Whorwell A, Clinicaltrial:theeffectsofafermentedmilkproductcontaining MorrisJ,ReillyB,GuyonnetD,Goupil-Feuillerat HoughtonL.A, N,Schlumberger Agrawal A, AGRAWAL over 4weeks. a nonfermenteddairyproductwithlowcontentoflactose(lactose similartothetestproduct)(n=20), EVALUATION CRITERIA consumeddaily2x125gservingsofeitherActivia Syndrome withpredominantconstipation(IBS-C) In thisrandomized,controlled,double-blind,parallelgroupstudy, 38women(18-70yo)withIrritableBowel STUDY METHODOLOGY consumption). evaluation atbaselineandafter4weeksofproduct marker andhydrogenbreathtestmethods; consumption). evaluation atbaselineandafter4weeksofproduct

Colonic andsmallboweltransittime(Radiopaque Abdominal distension(Plethysmography;24h distension atbaseline)/maximalbaselinex100. Percentage change inmaximaldistension=(maximaldistensionposttreatment- Data representsthemedian. Change in maximal distension (%) PERCENTAGE CHANGEINMAXIMALDISTENSIONAFTERACTIVIA -90 -75 -60 -45 -30 -15 0 et al.,2009 DN-173 010onabdominaldistensionandgastrointestinaltransitin OR CONTROLCONSUMPTION Control movement/week; dailyevaluation). bloating, ﬂ atulence; dailyevaluationreportedinadiary). symptoms severity(i.e.abdominalpain⁄discomfort, *

Stool consistency(BristolScale;dailyevaluation). Overall IBS symptoms severity and digestive symptomsseverityanddigestive Overall IBS Stool frequency(expressedasnumberofbowel Activia ® ® *p<0.05 ® (n=18)or 15/11/12 13:51

ACTIVIA® EFFICACY STUDIES: IBS-C POPULATION AGRAWAL et al., 2009

RESULTS Abdominal distension Overall IBS and digestive symptoms After 4 weeks of product consumption, a trend towards Over the 4 weeks of Activia® consumption, overall a reduction in mean abdominal distension during symptom severity signifi cantly improved (p=0.032), the day (AUC values, -1.52 cm 95% CI [3.33, 0.39], abdominal pain⁄discomfort signifi cantly decreased p=0.096) and a signifi cant reduction in the percentage (p=0.044), and bloating (p=0.059) and fl atulence change in maximal distension (Activia®: -77.1% vs. (p=0.092) tended to reduce, compared with control control: -28.6%, p<0.05) were observed in the Activia® group. group, in comparison with the control group. Stool consistency and frequency A trend to a normalization of stool consistency Colonic and small bowel transit time (p=0.058) was also observed without modifi cation Colonic and small bowel transit times were signifi cantly of stool frequency. reduced in the Activia® group, in comparison with the control group (respectively -12.2h, 95%CI [-22.8,-1.6], p=0.026 and -1.2h, 95%CI [-2.3, 0.0], p=0.049).

CONCLUSION Daily consumption of 2x125g servings of Activia® over 4 weeks, signiﬁ cantly reduced abdominal distension in association with acceleration of gastrointestinal transit in IBS-C women. Activia® improved digestive comfort by reducing overall IBS symptom severity and digestive symptoms.

MEAN ABDOMINAL DISTENSION BEFORE AND AFTER ACTIVIA® OR CONTROL CONSUMPTION

Before product consumption After product consumption 6 6

5 5

4 4

3 3

2 2 Mean distension (cm) 1 Mean distension (cm) 1

0 0 1 2 3 4 5 6 7 8 9 10 11 12 13 1 2 3 4 5 6 7 8 9 10 11 12 13 Hours Hours

Activia® group Control group Activia® group Control group

PDF ACTIVIA.indd 12 15/11/12 13:51 PDF ACTIVIA.indd 13 Therapeutics, 2007;26(3):475-486. Pharmacologyand care: amulticentre,randomized,double-blind,controlledtrial.Alimentary health-related qualityoflifeandsymptomsinirritablebowelsyndromeadultsprimary Effect ofafermentedmilkcontainingBiﬁ dobacterium animalisDN-173010onthe Guyonnet D,ChassanyO, DucrotteP, Picard C,MouretM,MercierC.H,MatuchanskyC. GUYONNET a heat-treated yoghurtcontainingnon-livingbacteria(n=132),over6weeks. EVALUATION CRITERIA consumeddaily2x125gservingsofeitherActivia Syndrome withpredominantconstipation(IBS-C) In thisrandomized,controlled,double-blind,parallelstudy, 267subjects(18-65yo)withIrritableBowel STUDY METHODOLOGY evaluation). movement/week; dailyevaluation). product consumption). evaluation atbaselineandweeks36of consumption). scale; evaluationafter3and6weeksofproduct 6 weeksofproductconsumption). discomfort scorevs.baseline;evaluationafter3and subjects havinganimprovement≥10%ofdigestive dimension ofFDDQLquestionnaire(deﬁ ned as weeks ofproductconsumption). FDDQL (Evaluationatbaselineandafter36 Digestive DisorderQualityofLifequestionnaire:

Stool consistency(BristolScale;daily Stool frequency(expressedasnumberofbowel Bloating andabdominalpain(6-pointsLikertscale; Global digestivesymptoms(7pointsLikert Rate ofrespondersforthedigestivediscomfort Digestive discomfortdimensionoftheFunctional et al.,2007 Activia improvement beingsignifi cantly (p=0.03)higherin in bothgroupsascomparedtobaseline,this Bloating scoresignifi cantly (p<0.001)improved Bloating andabdominalpain group (47.7%)(p=0.003). over 6weeksintheActivia week (n=19),stoolfrequencysignifi cantly increased In asubgroupofsubjectswithlessthan3stools⁄ Stool frequencyandconsistency difference betweengroups. in bothgroupsascomparedtobaselinewithout Abdominal painsignifi cantly (p<0.001)improved (p<0.001) inbothgroupsatweek3(Activia The FDDQLdigestivediscomfortscoreimproved Global digestivesymptoms RESULTS week 3intheActivia the FDDQLquestionnairewassignifi cantly higherat The responderrateforthediscomfortdimensionof Responders groups atweeks3and6. digestive symptomsscoresdidnotdifferbetween ± 16.2;control:13.519.3).Theevolutionofglobal 14.5; control:7.5±16.5)andweek6(Activia group (p<0.001). ® groupvs.controlatweek3. ® group(65.2%)vs.thecontrol ® groupvs.thecontrol ® (n=135)or ® : 10.7± ® : 12.2 15/11/12 13:51

ACTIVIA® EFFICACY STUDIES: IBS-C POPULATION GUYONNET et al., 2007

CONCLUSION Daily consumption of 2x125g servings of Activia® during 6 weeks improved digestive comfort. This study suggests a beneﬁ cial effect of Activia® on the FDDQL digestive discomfort score and bloating in subjects with IBS-C. A positive effect on stool frequency in subjects having less than 3 stools per week was also observed.

RATE OF RESPONDERS FOR DIGESTIVE COMFORT DIMENSION AFTER 3 AND 6 WEEKS OF ACTIVIA® OR CONTROL CONSUMPTION

17.5%* 6.2% *p=0.003 60

40 Activia® 30 Control 20 Responder rate (%) 10

0 Week 3 Week 6

BLOATING BEFORE AND AFTER 3 AND 6 WEEKS OF ACTIVIA® OR CONTROL CONSUMPTION

4 *p=0.03

3,5 Activia®

Score * Control 3

Score:1=no bloating 6=very important 2,5 Baseline Week 3 Week 6

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TRANSIT TIME & BOWEL MOVEMENT Yang et al., 2008 Nishida et al., 2008 Marteau et al., 2002

STRAIN SURVIVAL & GUT MICROBIOTA MODULATION McNulty et al., 2011 Rochet et al., 2008 Collado et al., 2006 Duez et al., 2000 Pochart et al., 1992 Berrada et al., 1991 ...... PDF ACTIVIA.indd 15 15/11/12 13:51 PDF ACTIVIA.indd 16 15/11/12 13:51 PDF ACTIVIA.indd 17 EVALUATION CRITERIA constipation) (25-65yo),consumeddaily100gofeitherActivia with constipation(lessthan3stools/week;increasedstoolhardness;non-organicandhabitual In thisrandomized,double-blind,placebo-controlled,parallelgroupstudy, 135healthyChinesewomen STUDY METHODOLOGY of Gastroenterology, 2008;14(40):6237-6243. containing Biﬁ dobacterium lactisDN-173010onChineseconstipatedwomen.World Journal Yang Y, HeM,HuG,Wei J,Pages P, Yang X,Bourdu-NaturelS.Effectofafermentedmilk YANG consumption). at baseline,andafter12weeksofproduct consumption). at baseline,andafter12weeksofproduct product consumption). evaluation atbaseline,andafter12weeksof non-living bacteria(n=68),over2weeks. Daily consumptionof100gActivia CONCLUSION defecation conditions,inChinesewomenwithconstipation.

Defecation condition(4gradescore,evaluation Stool consistency(Bristolstoolscale;evaluation Stool frequency(expressedasnumber/week; STOOL FREQUENCYAFTERONEORTWOWEEKSOFACTIVIA

Stool frequency (n/wk) et al.,2008 0 1 2 3 4 5 6 7 Baseline ® during2weeksimprovedstoolfrequencyandconsistencyaswell Week 1 ## ** Week 2 After 1and2weeksofActivia RESULTS group. week 2:p<0.05),incomparisontothecontrol consistency signifi cantly improved(week1:p<0.01, and defecationconditions(p<0.01)stool stool frequencysignifi cantly increased(p<0.01) ## ® ** (n=67)oranacidifi ed milkcontaining ® ##: intergroupcomparisonp<0.01 **: intragroupcomparisonp<0.01 ORCONTROLCONSUMPTION Activia Control ® ® consumption, 15/11/12 13:51

ACTIVIA® MECHANISM OF ACTION STUDIES: TRANSIT TIME & BOWEL MOVEMENT

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...... PDF ACTIVIA.indd 19 Cross-over StudyamongHealthyJapaneseWomen. Pharmacometrics2008;74(5/6):99-106. the ConditionofDefecationandIntestinalMicrofl ora: Arandomized,Double-blind,Placebo-controlled, Nishida S,IshikawaY, IinoH.EffectofBifidobacterium lactisDN-173010ontheIntestinalTransit Time, NISHIDA (n=25) beganwiththecontrolproductandthenconsumedActivia daily, thenconsumed2x85gservingsofacontrolproductwithoutB.lactisDN-173010daily. Thesecondgroup Daily consumptionof2x85gservingsActivia CONCLUSION age 19.43±1.62yo)weredividedin2groups.Thefi rst group(n=25)startedwith2x85gservingsofActivia In thisrandomized,double-blind,placebo-controlled,cross-overstudy, 50healthyJapanesewomen(mean STUDY METHODOLOGY EVALUATION CRITERIA of productconsumption).14subjects. counting, evaluationatbaseline,andafter2weeks 43 subjects. respectively, dailyevaluationrecordedinadiary). and shapedefi ned accordingtoscaleandmodel frequency expressedasnumberperweek;color consumption). 35subjects. evaluation atbaselineandafter2weeksofproduct period lasted2weekswitha6intervalbetweeningestionofeachproducts. increased thestoolfrequency, especiallyinJapanesewomenwithaslowtransittime.

Fecal microfl ora analysis(enumerationbyplate Stool frequency, quantity, colorandshape;(stool Oro-fecal transittime(Coloredmarkermethod, ORO-FECAL TRANSITTIMEBEFOREANDAFTERACTIVIA Sgi atydfeetcmae ihbsln p00) #p=0.055compared withcontrol *Signifi cantly differentcomparedwithbaseline(p<0.05) Values representthemean±S.D. Transit time (h) 100 25 50 75 0 et al.,2008 Baseline ControlActivia -9:55 -18:16 n=35 Total -8:21 * ® ® during2weeks,improvedtheintestinaltransittimeand Baseline ControlActivia Fast transittime -4:08 n=18 between the two groups. between thetwogroups. faeces, nosignifi cant differenceswereobserved to thequantity, colour, shapeandcharacteristicsof (>40 hours,17subjects),consumptionofActivia to thecontrolgroup.Insubjectswithslowtransittime (p<0.05) ascomparedwithbaselinebutnot signifi cantly reducedafterActivia In theoverallpopulation,intestinaltransittimewas RESULTS during theActivia ratio ofintestinalmicrofl ora signifi cantly increased The stool frequency(p<0.05)comparedtothecontrolgroup. tended toreducethetransittime(p=0.055)andincrease -8:48 -4:40 Bifi dobacterium cellcountandtheBifi dobacterium ® (samequantitiesdaily).Eachingestion ® ® ORCONTROLCONSUMPTION ® Baseline ControlActivia consumption (p<0.05). With regard regard consumption(p<0.05).With Slow transittime -24:48 -46:58 n=17 * -22:10 ® consumption consumption * # ® ®

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...... PDF ACTIVIA.indd 21 women: adouble-blind,randomized,controlledstudy. Pharmacol&Therap,2002;16:587-93. Alimentary Grimaud JC.Bifi dobacterium animalisstrainDN-173010shortensthecolonictransittimeinhealthy Marteau P, CuillerierE,MéanceS,GerhardtMF, BouvierM,BouleyC,Tondu MyaraA, F, BommelaerG, MARTEAU Daily consumptionof3x125gservingsActivia CONCLUSION This effectwasmorepronouncedinwomenwithinitialslowertransittime. Two valueshavingdifferentletterswithinasinglelinearestatistically signifi cantly different(p<0.05). Values: mean±SD ingestion ofeachproduct. EVALUATION CRITERIA divided intwogroups.Thefi rst group(n=17)startedwith3x125gservingsofActivia In thisrandomized,controlled,double-blind,multicenter, cross-overstudy, 32healthywomen(18-45yo)were STUDY METHODOLOGY Activia 3x125g servingsofayoghurtdaily. Thesecondgroup(n=15)beganwiththeyoghurtandthenconsumed ingestion period). evaluation atbaselineandaftereachproduct sigmoid (Radio-opaquemarkermethod, (Evaluation attheendofeachingestionperiod).

Transit Time(h) Colonic transittimes:totalright,leftand Fecal bilesalts,pH,microbialmassandweight Sigmoid Colonic ® (samequantitiesdaily).Eachingestionperiodlasted10days,withadaysintervalbetween COLONIC ANDSIGMOIDTRANSITTIMEINWHOLEPOPULATION ANDWOMEN 25.2 ±18.9 55.2 ±28.0 Baseline et al.,2002 Whole population(n=32) ab ab 21.6 ±14.9 51.5 ±30.2 WITH COLONIC TRANSITTIME>40H Activia ® a a 26.8 ±14.2 60.7 ±27.1 ® signiﬁ cantly reducedcolonictransittimeinwomen. Control the Activia times weresignificantly(p<0.05)shortenedin time >40hours,bothcolonicandsigmoidtransit In thesubgroupofsubjectswithaninitialtransit vs. baseline. control group,butnodifferencewasobserved shortened intheActivia sigmoid transittimesweresignificantly(p<0.05) In thetotalpopulation,bothcolonicand RESULTS were notsignificantlymodified. Fecal mass,pH,bacterialmassandbileacids baseline. b b 32.8 ±18.3 70.4 ±21.8 Subjects withcolonictransittime>40h(n=21) Baseline ® groupvs.thecontroland a a 27.1 ±14.9 62.4 ±29.8 ® Activia ® daily, thenconsumed group,comparedtothe ® b b 32.1 ±13.1 71.9 ±26.5 Control a a

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...... PDF ACTIVIA.indd 23 Science Translation Medicine,2011;3(106):106ra106. of fermentedmilkstrainsonthegutmicrobiomegnotobioticmiceandmonozygotictwins. GordonJ.I.Theimpactofaconsortium MuehlbauerM.J,NewgardC.B,HeathA.C, Bain J.R, DuncanA.E, KnightR, C,KnightsD,LozuponeC.A, Cools-PortierR, S,GobertG,Chervaux McNulty N.P, Yatsunenko T, Faith HenrissatB,Oozeer HsiaoA, J.J,MueggeB.D,GoodmanA.L, McNULTY Activia received either1x2gavageswithin24hor3x2(over3weeks)ofthe5Activia Human study: Human study: Animal model: EVALUATION CRITERIA Animal model: 2 approaches:a“humanized”animalmodelandcontrolledhumanstudy. STUDY METHODOLOGY After thedailyconsumptionof2x4ozservingsActivia CONCLUSION CFU intotalforeachgavage). pre-treatment, duringtreatment,post-treatment). (Metagenomics, Metatranscriptomics,evaluation during treatment,post-treatment). lactis infaeces(qPCR,evaluationpre-treatment, of theﬁ rst inoculation,post-treatment). produced?; 3measuresbeforecolonization,theday (Metabolomics =whatmoleculeshavebeen treatment). evaluation pre-treatment,duringtreatment,post- Metatranscriptomics =whatarebacteriadoing?; (Metagenomics =whichbacteriaarethere?; When healthypeopleconsumedActivia lactis CNCMI-2494wasdetectedthroughoutthehumangastrointestinal tract. the colon)remainedstable,butgeneexpressionof gutmicrobeswasmodulated.

Stool parametersandfrequency(daily). Sequencing technologiesonbacteriafromfaeces Quantifiof cation Mesurement ofmetabolitesinurinesamples Sequencing technologiesonbacteriafromfaeces ® during7weeks. 10 germ-freemice,colonizedwith15wellcharacterizedbacteriaoriginatedfromhumangut, 7 healthyfemaleadultmonozygotictwinpairsconsumeddaily2x4oz(=113g)servingsof Bifi dobacterium animalissubsp. et al.,2011 ® , theglobalcompositionofgutmicrobiota(bacterialiving in No detectionofsignifi cant impactofActivia limits ofdetectioninallparticipants. originate fromplantdietarycomponents.CMI-2 to utilizexylo-oligosaccharides(XOS) thatmainly I-2494 (DN-173010)isactiveinthegutandappears To endBifi dobacterium animalissubsp.lactisCNCM microbiota. the productionofshortchainfattyacidsby by themicrobiota. effi cacy ofutilizationplantderivedpolysaccharides to theweek7ofActivia Among the14healthyhumans,fromweek1 RESULTS modulated: Gene expressionofthegutmicrobeswas mice (nostatisticalsignifi cance). composition inhumans,andminormodifi cation in consumption onglobaldominantmicrobiota Activia was highinfeces.4weeksafterthecessationof Bifi dobacterium animalissubsp.lactisCNCMI-2494 ®

during7days,Biﬁ dobacterium animalissubsp. Activia Activia ® consumption,levelsdecreasedtobelowthe ® ® (productandstrains)couldpromote (productandstrains)couldpromotethe ® consumption,thelevelof ® strains(2.10 ®

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% Normalized counts Abundance of B. animalis subsp. lactis log10 (cell equivalents/g feces) 10 10 10 10 10 10 5.0 5.5 6.0 6.5 7.0 7.5 8.0 8.5 LEVELS OFBIFIDOBACTERIUMANIMALISSUBSP. INHUMANFECALSAMPLES LACTIS -6 -4.5 -4 -1.5 -3 -2.5 MOUSE ANDHUMANGUTMICROBIALCOMMUNITIESSHARETRANSCRIPTIONAL et al.,2011 Detection Limit Cellobiosephosphorylase d14 pre-treatment COLLECTED PRIORTO, DURINGANDAFTERCONSUMPTIONOFACTIVIA MOUSE Pre4 Pre2Pre1FMP1FMP2FMP4FMP7Post2 Post4 d15 treatment

d42 multi-treatment EC2.4.1.20 COMMISSION) RELATEDTOCARBOHYDRATEMETABOLISM d42 single-treatment RESPONSES TOACTIVIA

HUMAN Pre 1

Act 1 (qPCR, LOG Act 4 Post 4

d14 pre-treatment 10 MOUSE

d15 treatment (CELLEQUIVALENTS/gFECES)) Pectinesterase

d42 multi-treatment EC3.1.1.11 d42 single-treatment ® INVOLVING ECS(ENZYME

HUMAN Pre 1 Act 1 Act 4 Post 4

d14 pre-treatment MOUSE d15 treatment EC3.2.1.65

d42 multi-treatment Levanase d42 single-treatment Pre Activia® HUMAN Activia® (1w) Activia® (4w) Post Activia® ®

in expression Fold-Change Relative -10 10 -2 2 1 15/11/12 13:52 PDF ACTIVIA.indd 25 (n=6), over7days.Thefollowupperiodlasted10 Data arepresentedasCFU/goffaeces.UD:UnderDetection limit(<10 After thedailyconsumptionof3x125gservingsActivia CONCLUSION period of: consumption andattheendoffollow-up Evaluation atbaseline,theendofproduct EVALUATION CRITERIA (6.6.10 In thisrandomized,open,parallelstudy, 12healthysubjects(24-46yo)consumeddailyeither3x125gservings STUDY METHODOLOGY healthy adults.JournalofMolecularMicrobiologyandBiotechnology, 2008;14:128-136. administrationinlyophilisedformorfermentedproduct-Arandomizedstudy microbiota after ofBiﬁS, Picard C,Goupil-Feuillerat N,DoréJ.Survival dobacterium animalisDN-173010inthefecal Rochet V, BressonJ.L.,Cools AndrieuxC,SutrenM,RabotS,MogenetA, L,LedaireA, Rigottier-Gois ROCHET metabolites. Electrophoresis (PCR-TTGE). Reaction (PCR)-Temporal Temperature GradientGel Situ Hybridization(FISH)andPolymerase Chain faeces (Colonyimmunoblotting,FluorescentIn B. lactisDN-173010survivedpassagethroughthegastrointestinaltract.

Determination offecalenzymeactivitiesand Survival ofBifi dobacterium animalissp.lactisin Subject 11 LF Mean 10 LF 1 LF 8 LF 5 LF 4 LF 10 CFU/day)ofActivia QUANTIFICATION BY IMMUNODETECTION(colonyimmunoblotting)OF B. lactisDN-173010INFAECES ACTIVIA AFTER Day 0 UD UD UD UD UD UD Lyophilised form et al.,2008 ® (n=6)or1g(2.1.10 1.0 x10 4.5 x10 2.3 x10 3.9 x10 8.2x10 Day 7 >10 UD 9 8 8 7 9 8 Day 17 10 UD UD UD UD UD 11 5 CFU/day)offreeze-driedpowderB.lactisDN-173010 activities. of dominant membersofthefecalmicrobiotaortheir No majormodifi cations wereobservedineitherthe detected for11outof12subjects. B.lactisDN-173010patternswere PCR-TTGE, With days ofproductconsumption. faeces, in5outof6subjects,bothgroups,after7 B. animalissp.lactisDN-173010was≥10 immunoblotting method,themeannumberof According tothequantifi cation bythe RESULTS Subject 12 FP Mean 7 FP 9 FP 6 FP 3 FP 2 FP ® or1goffreeze-driedpowderduring10days, 4 CFU/gfaeces) Day 0 ® Fermented product UD UD UD UD UD UD CONSUMPTION 2.1 x10 1.9 x10 1.0 x10 1.1 x10 8.3x10 4.7x10 Day 7 UD 8 8 8 7 7 8 8 1.0x10 6.4x10 Day 17 CFU/gof UD UD UD UD 7 5 15/11/12 13:52

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...... PDF ACTIVIA.indd 27 After dailyconsumptionof250mlActivia CONCLUSION passage throughthegastrointestinaltract. Food ResearchInternational,2006;39:530-535. Bifianimalis DN-173010inhumanfecesduringfermentedmilkadministration. dobacterium Collado M.C,MorenoY, HernándezM.Moleculardetectionof CoboJ.M,MateosJ.A, EVALUATION CRITERIA daily250mlofActivia consumed In thisnonrandomized,controlled,opentrialincluding12healthyvolunteers(25-40yo),10subjects STUDY METHODOLOGY COLLADO of productconsumptionandfollowupperiods). (ADRA-PCR); evaluationatbaseline,andeachweek profi le (Amplifi ed RibosomalDNARestriction-PCR Chain Reaction(PCR)). product consumptionandfollowupperiods). (FISH); evaluationatbaseline,andeachweekof plate countingandFluorescentInSituHybridization weeks. subject ingesteddaily250mlofActivia

Identifiof cation Identifiof cation Detection ofbifi dobacteria genus(Enumerationby TOTAL BIFIDOBACTERIAGENUSCOUNTSAFTERCONSUMPTIONOFACTIVIA Sampling B. animalissp.lactisDN-173010 B. animalissp.lactis(Polymerase FISH counts (log cells/ml) 10 12 0 2 4 6 8 Control period et al.,2006 FISH countsBif662 1 ® (n=10)over4weeks,1subjectingestednoproduct(negativecontrol)and 2 345 Consumption period ® for3monthspriorstudy(positivecontrol).Thefollowupperiodlasted4 ® over4weeks,B.animalissp.lactisDN-173010survived Plate countsBFMagar was observed after 4 weeks follow-up period. was observedafter4weeksfollow-upperiod. consumption (90%after4weeks).Nomoredetection 010 strainbyARDRA-PCR,after2weeksofproduct 40% ofbifi dobacteria wereidentifi ed asDN-173 no moredetectableafter4weeksfollow-upperiod. consumption (90%after4weeks).Subspecieswere as studied period.30%ofbifi dobacteria wereidentifi ed and negativecontrolsremainedstable,overthe number ofbifi dobacteria genusinthepositive discontinuation oftheproduct.Incontrast, Count decreasedtobaselinelevel4weeksafter consumption genus infaeceswasobservedduringproduct A signifi cant increase(p<0.05)ofbifi dobacteria RESULTS 6 789 by PCR after 2 weeks of product B. lactissp.byPCRafter2weeksofproduct Follow-up period baseline. vs. baseline. 10 12 0 2 4 6 8

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...... PDF ACTIVIA.indd 29 After dailyconsumptionof3x125gservingsActivia CONCLUSION consumption ofhumanfecalsamples: 7daysofproduct Evaluation atbaselineandafter EVALUATION CRITERIA servings ofActivia In thisnonrandomized,controlled,openstudy, 5healthywomen,(20-48yo),consumeddaily3x125g STUDY METHODOLOGY probiotic straininhumanfaeces.JournalofAppliedMicrobiology, 2000;88:1019-27. A colonyimmunoblottingmethodforquatitativedetectionofaBifi dobacterium animalis Duez H,Pelletier C,CoolsS,AissiE,CayuelaGaviniF, BouqueletS,NeutC,MengaudJ. DUEZ specie). agar pH5,notincludingB.animalissp.lactis method specifi c tostrains). strains suchasDN-173010strain(immunoblotting (Beeren’s agarpH5.5,specifi c tothespecie). to beviable,inlargequantities(≥10

Quantifiof cation Population (log CFU/g) Quantifi cation ofseveralB.animalissp.lactis Quantifiof cation Before/after consumptionofActivia Bifi dobacterium animalissp.lactis(before/afterActivia 10 12 Bifiwithout dobacteria 0 2 4 6 8 Subject 1 ENUMERATION OFBIFIDOBACTERIAANDBifi dobacterium animalissp.lactis et al.,2000 ® Bifi dobacterium genus(Beeren’s B. animalissp.lactisspecie over7days. POPULATION INFECALSAMPLESOFFIVEHUMANVOLUNTEERS B. lactis(before/afterActivia BEFORE ANDAFTERONEWEEKINGESTIONOFACTIVIA Subject 2 ® 8 CFU/g),infaeces. ® ingestion),basedonBereen’sagarpH5method ® ingestion),basedonBeeren’sagarpH5.5andimmunoblottingmethods Subject 3 ® duringoneweek,B.lactisDN-173010wasfound amongst subjects,between10 fecal samplesbeforeingestionofActivia No coloniesofB.lactisweredetectedinanythe RESULTS days ofActivia ® Subject 4 consumption,B.lactiscountvaried ® 8 and10 Subject 5 9 CFU/g. ® . After7 n=5 15/11/12 13:52

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large quantityofingestedbifi dobacteria reachedthecolon. signifi cantly. After8hours,10 After consumptionof400gB.lactisDN-173010fermentedmilk,ilealfl ow ofbifi dobacteria increased CONCLUSION EVALUATION CRITERIA animalis sp.lactisDN-173010fermentedmilkoramonitoreddietcontainingnobifi dobacteria I STUDY METHODOLOGY AmericanJournalofClinicalNutrition,1992;55:78-80. study usingintestinalperfusion. ingested viafermentedmilkduringtheirpassagethroughthehumansmallintestine:aninvivo Pochart P, MarteauP, Bouhnik Y, GoderelI,BourliouxP, ofBifi RambaudJ.C.Survival dobacteria POCHART ingestion). in ilealperfusion,throughoutthe8hoursofproduct the terminalileum(enumerationbyplatecounting, n thisrandomized,controlled,openstudy, 6healthyvolunteers(18-30yo)consumedeither400gofBifi dobacterium

Quantiﬁ cation ofviablebiﬁ dobacteria reaching FLOW RATEOFBIFIDOBACTERIATHROUGHTHEILEUMAFTERINGESTION B. lactisDN-173010FERMENTEDMILKORCONTROLMEAL et al.,1992 9 CFUofthestrainwasrecoveredinterminalileum,indicatingthata

Log10 bacteria/hour 2 3 4 5 6 7 8 9 02468 CFU vs.10 remained low(1.6x10 of viablebifi dobacteria throughouttheexperiment bifi dobacteria wasobserved,andtheconcentration In thecontrolgroup,nomodifi cation ofthefl ow of RESULTS signifi cantly smallerthantheamountsingested(10 The meannumberofbifi dobacteria recoveredwas observed, after1.7±0.4hours. concentration of2.5x10 peak fl ow (6.3x10 was observedwithinonehourinallsubjects,anda signifi cant increaseintheilealfl ow ofbifi dobacteria In theB.lactisDN-173010fermentedmilkgroup,a 10 CFU, p<0.02). n=6 8 Time (hours) CFU/hour),correspondingtoa fermented milk B. lactisDN-173010 Control meal 5 CFU/L). 9 CFU/Lintheilealfl uid, was specie. 9

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...... PDF ACTIVIA.indd 33 B:12 healthyvolunteers(22-25yo)received250mlofeitherActivia Bifi dobacterium fermentedmilk). After consumptionof250gActivia CONCLUSION EVALUATION CRITERIA 10healthyvolunteers(20-45yo)received250gofeitherActivia A: emptying rate(B)werestudied. The studywasarandomized,controlled,double-blind,cross-overtrial.Bothinvivosurvival(A)andgastric STUDY METHODOLOGY hours). milk, byScintigraphy, every10minutes,over3 sulfur colloidsincorporatediningestedfermented 99m-Tc-technetium-labelled solutionofrhenium enumeration byplatecounting). and 30,6090minutesafteringestion; (Samples collectedusingagastrictubeimmediately Bifi dobacterium fermentedmilk). transit. milks: Survival during gastric transit. Journal of Dairy Science,1991;74:409-413. duringgastrictransit.JournalofDairy milks: Survival Berrada N,LemelandJ.F, LarocheG,ThouvenotP, Piaia M.Bifi dobacterium fromfermented BERRADA

B group:Gastricemptyingrate(detectionof A group:Survivalofbifi dobacterium strains et al.,1991 ® , Bifilactis DN-173010wasabletosurvivegastric dobacterium population inActivia After 90minutesofgastrictransit,Bifi dobacterium RESULTS emptying betweenActivia There wasnosignifi cant differenceingastric signifi(p<0.001). cant between thesurvivalofBifi dobacterium strainswas product decreasedby4logunits.Thedifference log units.Bifi dobacterium populationinthecontrol ® ® oracontrolproduct(anothercommercial oracontrolproduct(anothercommercial

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