Hintergrund: Gestillte Kinder haben im Vergleich zu nicht gestillten Kindern eine geringere Inzidenz von gastrointestinalen Infektionen und atopischen Erkrankungen. Man geht davon aus, dass der gesundheitsfördernde Effekt der Muttermilch teilweise über die intestinale Mikrobiota vermittelt wird. Diese ist in Stillkindern durch eine geringe Diversität und einen hohen Anteil an Bifidobakterien charakterisiert. Neueste Ansätze in der Weiterentwicklung industriell hergestellter Säuglingsnahrung zielen darauf ab, eine intestinale Mikrobiota zu fördern, die der von gestillten Kindern ähnelt. Die Supplementation von Säuglingsnahrung mit Probiotika (lebende Mikroorganismen) oder Präbiotika (unverdauliche Kohlenhydrate, die als Energiesubstrat für probiotische Bakterien dienen) könnte die bifidogene und antipathogene, aber auch immunmodulierende Wirkung der Muttermilch nachahmen. Aufgrund unterschiedlicher Interaktionen mit der Darmmikrobiota und dem Immunsystem fokussiert man mit der gleichzeitigen Gabe von Pro- und Präbiotika (Synbiotika) eine synergistische Wirkung an. Zielstellung und Studiendesign: In einer randomisiert-kontrollierten, klinischen Studie wurde untersucht, ob sich in den ersten drei Lebensmonaten von gesunden und termingerecht geborenen Kindern mit einer Synbiotikum-haltigen Säuglingsnahrung eine intestinale Mikrobiota etabliert, die der von gestillten Kindern gleicht. Das Synbiotikum setzte sich aus Bifidobacterium animalis ssp. lactis CNCM I 3446 (ältere Bezeichnung B. lactis BB-12) und Kuhmilcholigosacchariden zusammen. Die Studie umfasste zwei Gruppen von Kindern, die eine Säuglingsnahrung mit (SYN-Gruppe, n=21) oder ohne Supplement (KON-Gruppe, n=18) erhielten. Gestillte Kinder dienten als Referenz (REF-Gruppe, n=23). Um die Diversität der Bifidobakterien auf Speziesebene umfassend zu charakterisieren, wurden quantitative Real-Time PCR (qPCR)-Verfahren, basierend auf dem single-copy groEL als phylogenetisches Zielgen, zur spezifischen Quantifizierung von zwölf Bifidobakterienspezies in humanen Fäzes entwickelt und validiert. Ergebnisse: Die supplementierte Säuglingsnahrung war gut verträglich und unterstützte eine gesunde Entwicklung; vergleichbare anthropometrische Daten von SYN- und REF-Gruppe. Das Synbiotikum stimulierte selektiv das Wachstum von Laktobazillen und Bifidobakterien. Die Zellzahl für Laktobazillen der SYN-Gruppe war zur REF-Gruppe äquivalent (9,07±0,32 versus 9,90±0,27 log10 Zellen/g Fäzes TM [MW±SEM]; p<0,0019; Äquivalenzdifferenz von 1 log10 Zellen/g Fäzes TM) und höher als in der KON-Gruppe (8,27±0,31 log10 Zellen/g Fäzes TM [MW±SEM]). Die Zellzahl für Bifidobakterien war in der SYN-Gruppe am höchsten (11,54±0,05 versus 11,00±0,17 [REF-Gruppe] und 10,54±0,24 [KON-Gruppe] log10 Zellen/g Fäzes TM [MW±SEM]). In der SYN-Gruppe wurde die höchste Anzahl an Bifidobakterienspezies erfasst (167 mit [128 ohne] B. animalis in 56 Fäzesproben versus 98 und 93 in jeweils 51 Fäzesproben der REF- und KON-Gruppe). Neben Kinder-typischen Spezies wie B. bifidum und B. breve wurden auch Spezies, die für Erwachsene charakteristisch sind (B. adolescentis), häufiger in der SYN-Gruppe als in den Vergleichsgruppen nachgewiesen. Der pH-Wert in Fäzes von Kindern aus der SYN-Gruppe war niedriger als der aus der KON-Gruppe (6,07±0,20 versus 6,45±0,17 [MW±SEM]) und näher an dem von gestillten Kindern mit 5,29±0,12 (MW±SEM). Schlussfolgerung: Die Supplementation einer Säuglingsnahrung mit dem Synbiotikum aus CNCM I-3446 und Kuhmilcholigosacchariden führte zu einer Angleichung in der Zusammensetzung der intestinalen Mikrobiota und des fäkalen pH-Wertes an gestillte Kinder. Die in dieser Arbeit entwickelten groEL-basierten qPCR-Verfahren erlaubten eine spezifische und genaue Analyse der Bifidobakterienpopulation unter dem Einfluss eines Synbiotikums. / Background: Compared to formula-fed infants, breast-fed infants have a reduced incidence of gastrointestinal infections and atopic diseases. The health-promoting effect of breast milk is assumed to be partly mediated by the intestinal microbiota, which is characterized by a low diversity and a high proportion of bifidobacteria. Recent approaches in further development of infant formulae aim at promoting an intestinal microbiota similar to that of breast-fed infants. The supplementation of infant formula with probiotics (live microorganisms) or prebiotics (non-digestible carbohydrates, which serves as energy substrates for probiotic bacteria) could mimic the bifidogenic and antipathogenic, but also immunomodulating effect of breast milk. Due to various interactions with the gut microbiota and the immune system, the simultaneous administration of pro- and prebiotics (synbiotics) is focussed to have a synergistic effect. Objective and study design: In a randomized-controlled, clinical trial healthy full-term infants receiving an infant formula with synbiotic for the first three months of life were studied, whether an intestinal microbiota is induced, which is equivalent to that of breast-fed infants. The synbiotic consisted of Bifidobacterium animalis ssp. lactis CNCM I 3446 (previously known as B. lactis BB-12) and cow milk oligosaccharides. The study comprised two groups of infants receiving a starter formula with (SYN-group, n=21) or without supplement (KON-group, n=18). Breast-fed infants served as a reference (REF-group, n=23). In order to comprehensively characterize the bifidobacteria diversity at species level, quantitative real-time PCR (qPCR) assays based on the single-copy groEL as phylogenetic marker for the specific quantification of twelve bifidobacteria species in human feces were established and validated. Results: The supplemented formula was well tolerated and supported a healthy development; comparable anthropometric data of SYN- and REF-group. The synbiotic selectively stimulated the growth of lactobacilli and bifidobacteria. Lactobacilli levels were equivalent in SYN- and REF-group (9.07±0.32 versus 9.90±0.27 log10 cells/g feces DM [Mean±SEM]; p<0.0019; equivalence margin of 1 log10 cells/g feces DM) and higher than the KON-group (8.27±0.31 log10 cells/g feces DM [Mean±SEM]). The highest levels of bifidobacteria were observed in the SYN-group (11.54±0.05 versus 11.00±0.17 [REF-group] and 10.54±0.24 [KON-group] log10 cells/g feces DM [Mean±SEM]). The highest number of bifidobacteria species were obtained in the SYN-group (167 with [128 without] B. animalis in 56 fecal samples versus 98 and 93 in each of 51 fecal samples of the REF- and KON-group). Beside species, typically found in infants such as B. bifidum und B. breve, also species, which are characteristic for adults (B. adolescentis), were detected more often in the SYN-group than in the other study groups. Fecal pH was lower in the SYN- than in the KON-group 6.07±0.20 versus 6.45±0.17 [Mean±SEM]) and closer to that of breast-fed infants (5.29±0.12 [Mean±SEM]). Conclusion: In infants fed a starter formula supplemented with a synbiotic (CNCM I-3446 and cow milk oligosaccharides), composition of intestinal microbiota and fecal pH were closer to that of breast-fed infants. The groEL-based qPCR-assays, developed in this study, allowed a specific and accurate analysis of the bifidobacterial population in response to the synbiotic intake.

Nutrition of children of each age has its particular aspects. For the new born and infantile age it is mainly breast-feeding that is very significant. Nutrition with breast milk is ideally balanced, ensures the intake of high-quality nutrition that is well tolerated by the young organism and ensures the optimal growth and psychological development of the child. Thanks to its composition, breast milk increases the protective impact on the child with respect to infections and creates a natural bond between the mother and the child. In the Thesis, I was trying to look up decisive moments in the infantile nutrition concept both in this country and worldwide and to map the issue of breast feeding and formula feeding since the end of the Second World War up to the present. Also, I wanted to emphasize the irreplaceability of breast milk as wel as supplementing of artificial milks with probiotics, prebiotics, synbiotics and their importance in the nutrition of children. Objective No. 1 of the Thesis has been to look up the key points in the infantile nutrition concept in the world since the beginning of the nineteenth century and since 1945 until this day in the Czech Republic. Objective No. 2 has been to prove that infantile formula nutrition has been focusing also on the immunological aspect apart from the nutritional ones. Objective No. 3 has been to create and aducational program proposal, which can serve as a practical guide for the application of theoretical data in clinical practice. Data necessary to achieve the determined objectives have been acquired by studing, analyses and comparison of available written documents relating to the natural and formula nutrition of infants. Most of these documents have been intended for the educational purposes at secondary school and universities in the Czech Republic. Also, data heve been taken from publications for parents and broad public published since 1945 until this day. By way of illustration, certain charts containing schemes of artificial nutrition corresponding to the time of creation have been used. On these schemes I wanted to show the chages in the concepts brought about by the development not only in the knowledge and thinking of people dealing with the nutrition of infants but also in the new possibilities and technology of the food processing industry.

A link between the gut and the brain has been proposed to influence psychiatric disorders. Probiotics have been suggested to modify the gut microbiota and thereby improve autism symptoms in children. Attention deficit hyperactivity disorder (ADHD) has high comorbidity with other neuropsychiatric diagnoses, including autism. This is a follow-up of the first study examining a synbiotic intervention in patients with ADHD (Skott et al., 2019). In the original study, 114 adults participated. In this study, 38 adults were evaluated. The aim was to examine if suggested improvements remained 18 months post treatment. Specifically, if reductions were detected in comorbid autism symptoms, emotional dysregulation or functional impairment. The endpoints were measured using questionnaires: Autism-Spectrum Quotient (AQ), Difficulties in Emotion Regulation Scale (DERS-16) and Weiss Functional Impairment Rating Scale (WFIRS). No Synbiotic2000-specific effect was detected. Synbiotic2000 and placebo improved emotion regulation and life skill-functioning equally well. More research is needed to draw reliable conclusions. / En koppling mellan magen och hjärnan har i studier antytts påverka psykiatriska tillstånd. Probiotika har föreslagits förändra mag- och tarmkanalens bakterieflora och därigenom förbättra psykiatriska symtom hos barn med autism. ADHD har hög komorbiditet med andra neuropsykiatriska diagnoser, däribland autism. Detta är en långtidsuppföljning av RCT-studien som var först med att undersöka en synbiotika-intervention hos patienter med ADHD (Skott et al., 2019). I uppföljningen undersöktes 38 av de 114 vuxna som deltagit i originalstudien. Syftet var att undersöka om indikationerna till förbättring höll i sig 18 månader efter studieavslutet. Frågeställningen var om reduktion i komorbida autismsymtom, svårigheter med emotionsreglering eller funktionsnedsättning kunde identifieras. Detta undersöktes genom självskattningsskalor: Autism-Spectrum Quotient (AQ), Difficulties in Emotion Regulation Scale (DERS-16) och Weiss Functional Impairment Rating Scale (WFIRS). Ingen behandlingsspecifik effekt detekterades. Förbättringar av samma grad identifierades av Synbiotic2000 och placebo, utifrån emotionsreglering samt delskalan färdigheter. Mer forskning på området krävs för att möjliggöra tillförlitliga slutsatser. / BAMBA (Behandla Adhd med MagBakterier)