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Table S1. Specific primers and probes used for real-time PCR and Illumina HiSeq sequencing (V6 hypervariable region of the 16S rRNA gene). Table S2. Mock community analysis results. The initial composition of mock community (strains), phylotype identifications according to Greengenes (GG) and HOMD reference databases, and relative abundance of each phylotype are presented. Table S3. Differentially abundant OTUs between erythritol and control groups based zero-inflated Gaussian mixture model. A negative value for fold change (logFC) indicates an increase in the relative abundance of a particular OTU in the erythritol group compared to the control group. Table S4. Differentially abundant OTUs between erythritol and xylitol groups based zero-inflated Gaussian mixture model. A negative value for fold change (logFC) indicates an increase in the relative abundance of a particular OTU in the erythritol group compared to the xylitol group. Table S5. Differentially abundant genera between erythritol and sorbitol groups, and erythritol and xylitol groups based zero-inflated Gaussian mixture model. A negative value for fold change (logFC) indicates an increase of the relative abundance of a particular genus in the erythritol group compared to the sorbitol or xylitol group. Statistically significant changes are asterisked. Table S6. Details of molecular methods [34, 35, 37–44] (DOC 114 kb)

Väitekirja elektrooniline versioon ei sisalda publikatsioone., Hambakaaries on ülemaailmse levikuga haigus. Eestis on kaariese näitajad eriti laste hulgas siiani kõrged. Kaaries, mis on dünaamiline, pikaajaliselt kulgev protsess, vajab pikaajalisi ennetusmeetodeid. Üks võimalus hambakaariese riski vähendada on asendada kariogeensed süsivesikud vähem kariogeensete suhkuralkoholide ehk polüoolidega. Paljud eelnevad uuringud on näidanud ksülitooli tarvitamise edukust kaariese ennetamisel, kuid mitmete teiste polüoolide puhul ei ole nende ennetav efekt kinnitust saanud. Kliinilise uuringu eesmärgiks oli määrata kaariese levimus algklasside õpilastel ning võrrelda erinevate polüoolide (ksülitool, erütritool ja sorbitool kui positiivne kontroll) pikaajalise tarbimise mõju ja efektiivsust vahelduvas hammaskonnas. Uuring oli topeltpime, juhuvalimiga, kontrollitud, ettevaatav, kolmeaastalise sekkumisega (polüoolide tarvitamine) kliiniline uuring mis sisaldas uuritavate iga-aastast suuõõne kliinilist läbivaatust Tartu Ülikooli Stomatoloogia Kliinikus. Me leidsime, et nii kaariese esinemissagedus kui ka kaariese indeksid olid Kagu-Eesti algklasside õpilastel kõrged, mis sobitub eelnevate uuringutega ning need tõusid kiiresti, olles teise klassi õpilaste hulgas statistiliselt olulisemalt kõrgemad võrrelduna esimese klassi õpilastega – seega vajab antud eagrupp tõhusaid preventiivseid meetmeid. Alumised teised piimamolaarid ja esimesed jäävmolaarid olid kõige enam kahjustunud hambad vahelduvas hammaskonnas. Kaaries algab enamasti fissuuridest kohe hamba suhu lõikumise järgselt, mis oleks ennetatav silandi kasutamisega, kuid läbivaadatud lastel oli silantide hulk väga madal. Kuna eelnevad uuringud on kinnitanud ksülitooli kaariest vähendavat efekti, on antud uuringu puhul ksülitooli tagasihoidlikku mõju raske seletada. Üks võimalik selgitus võib olla vähene tarvitamiskordade arv, mis oli kolm korda koolipäeva jooksul üldiselt soovitatava 4-5 päevale jaotatud tarvitamiskorra asemel. Kaariese areng oli erütritooli grupis aeglasem kui sorbitooli ja ksülitooli gruppides ning avaldus pärast 2 aastast erütritooli pastillide tarvitamist. Erütritool vähendas võrreldes sorbitooli ja ksülitooli pastillidega ühtlasi hambakatu hulka, hapete taset hambakatus ja Streptococcus mutans´i taset. Kokkuvõtvalt võime öelda et erütritool kui hästi talutav magusaine omab hambakatu hulga ja omaduste mõjutamise kaudu kõrget potentsiaali kaariese vähendamisel. Uuringu tulemuste põhjal võib erütritooli pastille soovitada asendamaks suhkruid maiustustes, et ennetada hambakaariest lastel., Dental caries is a globally occurring disease. The caries index in Estonia, especially among children, is still high. Caries is a dynamic, prolonged process, and therefore, needs long-term caries prevention methods. The replacement of cariogenic sugars with non-cariogenic substances, like xylitol or other less cariogenic sugar alcohols, is one way to reduce the risk of caries. Previous studies about the effect of xylitol on caries prevention have demonstrated evidence-based success but the preventive effect of other polyols has not always been confirmed or approved. The interest of this clinical trial was to identify the prevalence of caries among elementary school level children and to investigate and compare the influence and effectiveness of the long-term consumption of different polyols (xylitol, erythritol and sorbitol as a positive control) on mixed dentition. The study was designed as a double-blind randomised controlled prospective clinical trial with 3-year intervention (polyols consumption) which included annual clinical examination of the participants in the Department of Stomatology at the University of Tartu. We found that the prevalence and experience of caries among elementary school children in south-eastern Estonia is high (which matches previous studies) and increases rapidly - there is a statistically significant difference between first and the second graders, this age group needs intensive caries prevention programmes. Lower second primary molars and the first permanent molars are the most frequently affected teeth in mixed dentition. Dental caries starts in fissures, predominantly shortly after the eruption of the teeth which may be prevented using sealants, but the number of sealants among the examined children is extremely low. As previous studies have proven the caries decreasing effect of xylitol, the modest role of xylitol in this study is difficult to explain. One probable explanation could be insufficient consumption per day – 3 times per school day against 4–5 regular, daily intake, which has been generally recommended. Caries progress was slower in the erythritol group compared to the sorbitol and xylitol groups and the effect was apparent after 2 years of erythritol lozenge consumption. Erythritol consistently reduces the amount of dental plaque, the levels of acids in dental plaque and the counts of Streptococcus mutans compared with sorbitol and xylitol lozenges. We can conclude that erythritol is a well tolerated sweetener which has high potential in caries reduction through controlling plaque quality and quantity. Based on the results obtained, erythritol based lozenges could be recommended to replace sugar containing candies to prevent dental caries in children., http://www.ester.ee/record=b4461126*est