<p>Recent years there has been a large increase in the worldwide sales of cultured products containing probiotics. The obstacle in the production of fermented products incubated with probiotic strains is its effect on sensory, chemical and physical properties. This Master´s Thesis investigates how extended storage times in fermentation tank as well as buffer tank during production of probiotic yoghurts affect final product quality. Through experimental studies, maximum storage times for probiotic yoghurts were to be recommended. The study includes analyses of pH, viscosity, stability, acetic acid concentration, microbial viability and sensory evaluations.</p><p>Design of experiments (DOE) was used to construct a 3²-factorial design for the experiment where different storage times were tested. The analyses showed that:</p><ul><li>pH decreases with both storage time in fermentation tank and buffer tank.</li><li>Viscosity increases with long storage times in fermentation tank, while it decreases with long storage times in buffer tanks.</li><li>Many samples had a sandy/grainy and slimy texture which could be related to high levels of Bifidobacteria or high fermentation temperatures.</li><li>High levels of acetic acid was found in all samples.</li><li>All measurements data where within the specification limits, whereas no recommendation for maximum storage times during production could be given.</li></ul>

<p>Probiotika, nyttiga bakterier som är välgörande för människan, förekommer allt mer i olika produkter riktade till små barn – alltifrån droppar och kapslar till modersmjölksersättningar och grötar. Inom barnhälsovården saknar barnsjuksköterskan/ distriktssköterskan vägledning och riktlinjer angående probiotika i små barns kost. Syftet med litteraturstudien var att undersöka evidens angående probiotikas hälsoeffekter i preventivt syfte på friska små barn. Metoden var en litteraturstudie baserad på 20 vetenskapliga artiklar. Analysen resulterade i att probiotika var säkert att ge till friska små barn. Samtliga studier gav enhetligt resultat, att inga nackdelar fanns med att ge små barn probiotika. Inga negativa hälsoeffekter med probiotika kom fram i denna litteraturstudie. Positiva hälsoeffekter av stor betydelse fick barn som erhållit probiotika vid kolik, då snabb lindring inom en vecka framkom. Ett övervägande positivt resultat sågs vid användning av probiotika till små barn för att förebygga eksem. Probiotika minskade även frånvaron från dagis då infektionssjukdomar minskade hos barnen. Antibiotikaförskrivningen sjönk också hos de förskolebarn som fått probiotika. Probiotikas positiva hälsoeffekter kan användas till rådgivning och rekommenderas till små barn av barnsjuksköterskan/ distriktssköterskan inom barnhälsovården. Fördjupad forskning är önskvärd då utökad eller kombinationer av flera probiotiska bakteriestammar kan ge utökade hälsoeffekter.</p> / <p>Probiotics, beneficial and useful bacterias for the human body, occur more often in different products targeted to small children – in everything from drops and capsules to formula and porridge. Within the child health care, the pediatric- and the district nurse lacks guidance and guidelines concerning probiotics in small children's diet. The aim with the literature review was to examine evidence concerning probiotics health effects in prevention, in small children. The method was a literature review based on 20 scientific articles. The analysis resulted in that probiotics were safe to give to healthy small children. All studies gave uniformly results and showed no disadvantages with giving small children probiotics. No negative health effects were detected in this literature review. Positive health effects of great importance have shown be beneficial within treatment of colic with probiotics given to infants since relief occurred within a week. A predominately positive result was seen in use of probiotics to small children in order to prevent eczemas came to light. Probiotics also decreased absence from day care centers since infection diseases were reduced among the children. Antibiotic prescriptions also decreased when probiotics were used to pre-school children. The positive health effects of probiotics can be used for health recommendations to small children within the child health care by the pediatric- and the district nurse. Extended research is desirable since increased or and combinations of several probiotic bacterium stutters can give increased health effects.</p>

Probiotika, nyttiga bakterier som är välgörande för människan, förekommer allt mer i olika produkter riktade till små barn – alltifrån droppar och kapslar till modersmjölksersättningar och grötar. Inom barnhälsovården saknar barnsjuksköterskan/ distriktssköterskan vägledning och riktlinjer angående probiotika i små barns kost. Syftet med litteraturstudien var att undersöka evidens angående probiotikas hälsoeffekter i preventivt syfte på friska små barn. Metoden var en litteraturstudie baserad på 20 vetenskapliga artiklar. Analysen resulterade i att probiotika var säkert att ge till friska små barn. Samtliga studier gav enhetligt resultat, att inga nackdelar fanns med att ge små barn probiotika. Inga negativa hälsoeffekter med probiotika kom fram i denna litteraturstudie. Positiva hälsoeffekter av stor betydelse fick barn som erhållit probiotika vid kolik, då snabb lindring inom en vecka framkom. Ett övervägande positivt resultat sågs vid användning av probiotika till små barn för att förebygga eksem. Probiotika minskade även frånvaron från dagis då infektionssjukdomar minskade hos barnen. Antibiotikaförskrivningen sjönk också hos de förskolebarn som fått probiotika. Probiotikas positiva hälsoeffekter kan användas till rådgivning och rekommenderas till små barn av barnsjuksköterskan/ distriktssköterskan inom barnhälsovården. Fördjupad forskning är önskvärd då utökad eller kombinationer av flera probiotiska bakteriestammar kan ge utökade hälsoeffekter. / Probiotics, beneficial and useful bacterias for the human body, occur more often in different products targeted to small children – in everything from drops and capsules to formula and porridge. Within the child health care, the pediatric- and the district nurse lacks guidance and guidelines concerning probiotics in small children's diet. The aim with the literature review was to examine evidence concerning probiotics health effects in prevention, in small children. The method was a literature review based on 20 scientific articles. The analysis resulted in that probiotics were safe to give to healthy small children. All studies gave uniformly results and showed no disadvantages with giving small children probiotics. No negative health effects were detected in this literature review. Positive health effects of great importance have shown be beneficial within treatment of colic with probiotics given to infants since relief occurred within a week. A predominately positive result was seen in use of probiotics to small children in order to prevent eczemas came to light. Probiotics also decreased absence from day care centers since infection diseases were reduced among the children. Antibiotic prescriptions also decreased when probiotics were used to pre-school children. The positive health effects of probiotics can be used for health recommendations to small children within the child health care by the pediatric- and the district nurse. Extended research is desirable since increased or and combinations of several probiotic bacterium stutters can give increased health effects.

The human vaginal ecosystem is dominated by Lactobacillus species. An altered vaginal flora can result in symptomatic conditions such as bacterial vaginosis and vulvo-vaginal candidiasis, and urogenital colonisation by uropathogenic bacteria can cause urinary tract infection. The protective role of lactobacilli is gradually being accepted and clinical studies have been carried out in order to evaluate the use of promising probiotic bacteria, which are defined as “live microorganisms which when administered in adequate amounts confer a health benefit on the host”. This thesis includes an investigation into the ecological role of lactobacilli in the genital tract in healthy women, with respect to the relationship to other species and vaginal pH. Furthermore, in order to find different probiotic strains with promising probiotic qualities, Lactobacillus strains were screened in two diverse screening processes. The selected strains were further evaluated in clinical trials. The prevalence of group B streptococci (GBS) and yeast was significantly dependent on the number of vaginal lactobacilli among healthy women. GBS were less frequently found in women with high numbers of vaginal lactobacilli than in women with low numbers and the prevalence of yeast was significantly higher in women with 3-6.99 log10 lactobacilli sample-1 than in women with less than 3 or ≥7 log10 lactobacilli sample-1. Furthermore, the first screening made on 511 strains isolated from the female genital tract resulted in the final selection of a Lactobacillus plantarum, designated LB931. The screening showed that LB931 had a strong technical growth, survived through freeze-thawing, produced substances bactericidal to uropathogenic bacteria and was a rapid and strong producer of hydrogen peroxide. Further characterisation showed that LB931 possessed the properties required for probiotics with the capability to prevent urogenital infections. LB931 could be supplied to the genital tract through the usage of panty liners impregnated with the strain. In the second screening, Lactobacillus fermentum, designated Ess-1, was the only one out of 126 Lactobacillus strains with strong capacity to inhibit Candida albicans and Candida glabrata. Additional characterisation showed that L. fermentum Ess-1 had the properties that are needed to prevent over-growth of Candida in the vulvo-vaginal tract. The result of the case study showed that a high and frequent dosage of Ess-1 is needed and that improved vulvo-vaginal candidiasis specific diagnostic criteria are required. In conclusion, L. plantarum LB931 and L. fermentum Ess-1 are promising probiotic strains to be used in the prevention of recurrent urogenital infections in women and to enhance the normal flora in healthy women.

Background: Increased numbers of intestinal E. coli are observed in inflammatory bowel disease, but the reasons for this proliferation and it exact role in intestinal inflammation are unknown. Aim of this PhD-project was to identify E. coli proteins involved in E. coli’s adaptation to the inflammatory conditions in the gut and to investigate whether these factors affect the host. Furthermore, the molecular basis for strain-specific differences between probiotic and harmful E. coli in their response to intestinal inflammation was investigated. Methods: Using mice monoassociated either with the adherent-invasive E. coli (AIEC) strain UNC or the probiotic E. coli Nissle, two different mouse models of intestinal inflammation were analysed: On the one hand, severe inflammation was induced by treating mice with 3.5% dextran sodium sulphate (DSS). On the other hand, a very mild intestinal inflammation was generated by associating interleukin 10-deficient (IL-10-/-) mice with E. coli. Differentially expressed proteins in the E. coli strains collected from caecal contents of these mice were identified by two-dimensional fluorescence difference gel electrophoresis. Results DSS-experiment: All DSS-treated mice revealed signs of a moderate caecal and a severe colonic inflammation. However, mice monoassociated with E. coli Nissle were less affected. In both E. coli strains, acute inflammation led to a downregulation of pathways involved in carbohydrate breakdown and energy generation. Accordingly, DSS-treated mice had lower caecal concentrations of bacterial fermentation products than the control mice. Differentially expressed proteins also included the Fe-S cluster repair protein NfuA, the tryptophanase TnaA, and the uncharacterised protein YggE. NfuA was upregulated nearly 3-fold in both E. coli strains after DSS administration. Reactive oxygen species produced during intestinal inflammation damage Fe-S clusters and thereby lead to an inactivation of Fe-S proteins. In vitro data indicated that the repair of Fe-S proteins by NfuA is a central mechanism in E. coli to survive oxidative stress. Expression of YggE, which has been reported to reduce the intracellular level of reactive oxygen species, was 4- to 8-fold higher in E. coli Nissle than in E. coli UNC under control and inflammatory conditions. In vitro growth experiments confirmed these results, indicating that E. coli Nissle is better equipped to cope with oxidative stress than E. coli UNC. Additionally, E. coli Nissle isolated from DSS-treated and control mice had TnaA levels 4- to 7-fold higher than E. coli UNC. In turn, caecal indole concentrations resulting from cleavage of tryptophan by TnaA were higher in E. coli Nissle- associated control mice than in the respective mice associated with E. coli UNC. Because of its anti-inflammatory effect, indole is hypothesised to be involved in the extension of the remission phase in ulcerative colitis described for E. coli Nissle. Results IL-10-/--experiment: Only IL-10-/- mice monoassociated with E. coli UNC for 8 weeks exhibited signs of a very mild caecal inflammation. In agreement with this weak inflammation, the variations in the bacterial proteome were small. Similar to the DSS-experiment, proteins downregulated by inflammation belong mainly to the central energy metabolism. In contrast to the DSS-experiment, no upregulation of chaperone proteins and NfuA were observed, indicating that these are strategies to overcome adverse effects of strong intestinal inflammation. The inhibitor of vertebrate C-type lysozyme, Ivy, was 2- to 3-fold upregulated on mRNA and protein level in E. coli Nissle in comparison to E. coli UNC isolated from IL-10-/- mice. By overexpressing ivy, it was demonstrated in vitro that Ivy contributes to a higher lysozyme resistance observed for E. coli Nissle, supporting the role of Ivy as a potential fitness factor in this E. coli strain. Conclusions: The results of this PhD-study demonstrate that intestinal bacteria sense even minimal changes in the health status of the host. While some bacterial adaptations to the inflammatory conditions are equal in response to strong and mild intestinal inflammation, other reactions are unique to a specific disease state. In addition, probiotic and colitogenic E. coli differ in their response to the intestinal inflammation and thereby may influence the host in different ways. / Hintergrund: Chronisch entzündliche Darmerkrankungen zeichnen sich unter anderem durch eine starke Proliferation intestinaler E. coli aus. Unbekannt ist jedoch, ob diese Vermehrung eine Ursache oder eine Folge der Erkrankung darstellt. Ziel der vorliegenden Doktorarbeit war es daher, E. coli-Proteine zu identifizieren, welche der Anpassung an die entzündlichen Bedingungen im Darmtrakt dienen und unter Umständen einen Effekt auf den Gesundheitszustand des Wirtes haben. Weiterhin sollten die molekularen Ursachen für stammesspezifische Unterschiede zwischen probiotischen und gesundheitsschädlichen E. coli näher untersucht werden. Methoden: In den tierexperimentellen Analysen wurden keimfreie Mäuse entweder mit dem probiotischen E. coli Nissle oder dem adhärent-invasiven E. coli UNC monoassoziiert und in zwei verschiedenen Entzündungsmodellen näher untersucht. Einerseits wurde eine starke Darmentzündung durch die Gabe von 3,5% Natrium-Dextransulfat (DSS) ausgelöst. Andererseits wurde in Interleukin 10-defizienten (IL-10-/-) Mäusen eine sehr milde Form der Entzündung durch Besiedlung mit E. coli induziert. Die E. coli Bakterien wurden am Ende der Versuche aus den Caecuminhalten der Mäuse isoliert und die bakterielle Proteinexpression wurde mittels zwei-dimensionaler Gelelektrophorese analysiert. Ergebnisse des DSS-Versuchs: Alle Tiere des DSS-Versuchs entwickelten unabhängig vom E. coli Stamm, mit dem sie besiedelt waren, eine moderate Entzündung im Caecum und eine starke im Colon, wobei die Entzündungsreaktion durch die Monoassoziation mit E. coli Nissle leicht abgeschwächt wurde. In beiden E. coli Stämmen führte die Darmentzündung zu einer verringerten Expression von Enzymen des Kohlenhydratabbaus und der Energiegewinnung. In Folge dessen waren die intestinalen Konzentrationen bakterieller Fermentationsprodukte in den entzündeten Tieren geringer als in den gesunden Kontrolltieren. Weitere differentiell exprimierte Proteine umfassen das Fe-S- Cluster Reparaturprotein NfuA, die Tryptophanase TnaA und das uncharakterisierte Protein YggE. In beiden E. coli Stämmen, welche aus den DSS-Tieren isoliert wurden, war das NfuA Protein dreifach höher exprimiert. Eine Darmentzündung führt zu einer vermehrten Bildung reaktiver Sauerstoffspezies, welche die Fe-S-Cluster in Eisen-Schwefel-Proteinen zerstören und damit zu einer Inaktivierung dieser Proteine führen. In vitro Untersuchungen bestätigten, dass die Reparatur der Eisen-Schwefel-Proteine durch NfuA ein wichtiger Mechanismus ist um oxidativem Stress entgegenzuwirken. Das YggE Protein, welches laut Literaturangaben einen hemmenden Einfluss auf die Bildung reaktiver Sauerstoffspezies hat, war in E. coli Nissle 4- bis 8-fach erhöht (verglichen mit E. coli UNC unter Kontroll- und Entzündungsbedingungen). In vitro Versuche bestätigten diese Daten und zeigten, dass E. coli Nissle im Vergleich zu E. coli UNC eine erhöhte Resistenz gegenüber oxidativem Stress aufweist. Außerdem wurde im Vergleich E. coli Nissle vs. E. coli UNC (unter Entzündungs- und Kontrollbedingungen) ein 4- bis 7-fach erhöhter TnaA-Gehalt nachgewiesen. Indol, das Produkt der TnaA-katalysierten Tryptophanspaltung wurde in erhöhten Mengen im Intestinaltrakt E. coli Nissle-assoziierter Kontrolltiere detektiert. Seit längerem werden entzündungshemmende Eigenschaften für Indol postuliert, die aufgrund der Ergebnisse dieser Doktorarbeit nun auch mit den gesundheitsfördenden Eigenschaften von E. coli Nissle in Zusammenhang gebracht werden können. Ergebnisse des IL-10-/-- Versuchs: Nach einer 8-wöchigen Assoziationsdauer wurde nur in den mit E. coli UNC besiedelten IL-10-/- Tieren eine schwache Entzündungsreaktion nachgewiesen. Bedingt durch diese sehr schwach ausgeprägte Entzündungsantwort waren auch die Veränderungen im bakteriellen Proteom von E. coli UNC nur gering. Wie im DSS-Versuch waren Proteine des bakteriellen Energiestoffwechsels reprimiert, allerdings wurde keine Induktion von NfuA beobachtet. Daher scheint die Induktion von NfuA nur der Anpassung an eine starke Entzündung zu dienen. Weiterhin wurde nachgewiesen, dass E. coli Nissle aus IL-10-/- Tieren den Hemmer für das vertebrate C-Typ Lysozym (Ivy) sowohl auf mRNA- als auch auf Proteinebene stärker exprimiert als E. coli UNC. Überexpression von Ivy unter in vitro Bedingungen zeigte, dass es an der erhöhten Lysozymresistenz von E. coli Nissle beteiligt ist und somit eine Rolle als möglicher Fitnessfaktor von E. coli Nissle spielt. Schlussfolgerungen: In dieser Doktorarbeit wurde gezeigt, dass Darmentzündungen die Proteinexpression eines im Darm lebenden Bakteriums beeinflussen. Einige der aufgedeckten bakteriellen Anpassungsreaktionen werden sowohl bei einer starken als auch bei einer schwachen Entzündung ausgelöst; andere wiederum sind spezifisch für nur einen dieser Entzündungszustände. Weiterhin wurde deutlich, dass sich E. coli-Stämme hinsichtlich ihrer Reaktion auf eine Darmentzündung unterscheiden und damit möglicherweise den Wirt beeinflussen. 

Recent years there has been a large increase in the worldwide sales of cultured products containing probiotics. The obstacle in the production of fermented products incubated with probiotic strains is its effect on sensory, chemical and physical properties. This Master´s Thesis investigates how extended storage times in fermentation tank as well as buffer tank during production of probiotic yoghurts affect final product quality. Through experimental studies, maximum storage times for probiotic yoghurts were to be recommended. The study includes analyses of pH, viscosity, stability, acetic acid concentration, microbial viability and sensory evaluations. Design of experiments (DOE) was used to construct a 32-factorial design for the experiment where different storage times were tested. The analyses showed that: pH decreases with both storage time in fermentation tank and buffer tank. Viscosity increases with long storage times in fermentation tank, while it decreases with long storage times in buffer tanks. Many samples had a sandy/grainy and slimy texture which could be related to high levels of Bifidobacteria or high fermentation temperatures. High levels of acetic acid was found in all samples. All measurements data where within the specification limits, whereas no recommendation for maximum storage times during production could be given.

Probiotics are viable microorganisms that show beneficial effects on the health of the host by improving their intestinal microflora. The microorganisms applied as probiotics mainly include Lactobacillus and Bifidobacterium species. Probiotics can inhibit the bacterial pathogens, reduce serum cholesterol levels, improve lactose tolerance and stimulate the immune response. They also have other properties such as / tolerance to acid and bile salts, adherence to gastrointestinal cells for colonization, resistance to antibiotics and &beta / -galactosidase acitivity. The properties of probiotic products are determined by the characteristics of the microorganisms they contain. For that reason, isolation and characterization of new strains having probiotic properties is an important issue. New strains are generally isolated from their natural habitats which are fermented dairy products such as kefir. In order to exert beneficial health affects in the digestive system, commercial probiotic products should contain adequate numbers of viable cells. Probiotic microorganisms should protect their viability during their shelf storage. Therefore, the viability of probiotics is especially important for food manufacturers that search for new probiotic strains with good survival and stability properties upon storage. In this study, probiotic microorganisms were isolated from traditional kefir grains known as a &lsquo / complex probiotic&rsquo / . The isolates were firstly identified using biochemical tests, then the putative species belonging to &lsquo / Lactobacillus acidophilus group&rsquo / were identified with 16S rRNA gene sequencing. Analysis of sequencing resulted in differentiation of &ldquo / L. acidophilus group&rdquo / organisms, namely L. amylovorus and L. acidophilus. Moreover, typing of commercial and traditional L. acidophilus strains and L. amylovorus strains were performed with RAPD-PCR by using primer M13. While several L. acidophilus strains showed different RAPD fingerprints most of the L. acidophilus and L. amylovorus strains could not be differentiated due to high similarity of their RAPD fingerprints. Following identification, survival of these isolates in probiotic yogurt preparations were investigated and compared to the survival of commercial probiotics. Consequently, although the survival of kefir grain isolates were less than commercial probiotics, they sustained the minimum recommended level for probiotics (106 cfu/ml) during cold storage. Such level of survival makes them considerably good candidates to be used as commercial probiotic cultures.

Zusammenfassung Untersuchungen zur Beeinflussung von experimentellen Salmonella Enteritidis PT4 Infektionen bei Küken durch den probiotischen Stamm Escherichia coli Nissle 1917 Imad Azmi Awadein Mohamed Institut für Bakteriologie und Mykologie, Veterinärmedizinische Fakultät, Universität Leipzig und Institut für Geflügelkrankheiten, Veterinärmedizinische Fakultät, Freie Universität Berlin Eingereicht im Januar, 2004 Schlüsselworte: (E. coli Stamm Nissle 1917, Küken, S. Enteritidis, Probiotika, Bdellovibrionen) (94 Seiten, 48 Abbildungen, 16 Tabellen und 150 Referenzen) 1- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Stamm Nissle 1917 und Bdellovibrio bacteriovorus auf die Reduktion der Ansiedlung von S. Enteritidis PT4, auf die Reduktion der Mortalitätsrate, auf die Gesamtzahl aerober und Gram-negativer Bakterien, auf die Anzahl von Bdellovibrionen und auf das Körpergewicht von SPF-Hühnern. Im Vergleich zur positiven Kontrollgruppe konnten bei der Ansiedlung von S. Enteritidis PT4 im Caecum der Testgruppen an den Tagen 14, 21 und 28 signifikante Unterschiede festgestellt werden. Im Vergleich zur positiven Kontrollgruppe, die eine Mortalitätsrate von 10% aufweist, war die Mortalitätsrate der Testgruppen deutlich reduziert. Ebenso waren in den Testgruppen die aerobe und Gram-negative Gesamtkeimzahl am Tag 28 im Vergleich zur positiven Kontrollgruppe deutlich verringert. Signifikanten Anstieg der Bdellovibrionen Anzahl zeigten die Testgruppen sowohl im Vergleich zur positiven als auch negativen Kontrollgruppe. Beim Vergleich zwischen dem Gesamtkörpergewicht der Testgruppen zu dem der negativen und den überlebenden Tieren der positiven Kontrollgruppe gab es keine signifikanten Unterschiede. 2- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Stamm Nissle 1917 und Bdellovibrio bacteriovorus auf die Reisolation von S. Enteritidis PT4 aus verschiedenen Teilen des Intestinaltrakts und diverser Organe von SPF-Hühnern. In Kropf, Proventrikel, Herzblut, Lunge, Leber und Milz der Testgruppen konnte im Vergleich zur positiven Kontrollgruppe eine Reduktion ermittelt werden, die Reisolation von S. Enteritidis aus dem Caecum der Testgruppen zeigte keinen Unterschied im Vergleich zur positiven Kontrollgruppe. 3- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Stamm Nissle 1917 und Bdellovibrio bacteriovorus an Konzentrationen von Serum-Avidin, IgY gegen S. Enteritidis und IgY gegen E. coli Nissle 1917 bei experimentell mit S. Enteritidis-infizierten SPF-Hühnern. Die Untersuchungen ergaben eine signifikante Senkung der Serum-Avidin-Konzentration im Vergleich zur positiven Kontrollgruppe, vor allem sieben Tage post infectionem mit S. Enteritidis. Ein signifikanter Anstieg der Konzentration von IgY gegen S. Enteritidis konnte bei den Testgruppen im Vergleich zur positiven Kontrollgruppe vor allem an den Tagen sieben verzeichnet werden. Ein signifikanter Anstieg der Konzentration von IgY gegen E. coli Nissle 1917 konnte in allen Testgruppen im Vergleich zur positiven Kontrollgruppe festgestellt werden. 4- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Nissle 1917 mit und ohne Plasmid auf die Reduktion der Ansiedelung von S. Enteritidis PT4, auf die Reduktion der Mortalitätsrate, die aerobe und Gram-negative Gesamtkeimzahl, die Bdellovibrionen-Gesamtkeimzahl, Keimzahlen von Lactobacillus, Bifidobacterium und C. perfringens und das Körpergewicht von SPF-Hühnern. Im Vergleich zur positiven Kontrollgruppe konnte im Caecum der Testgruppen ein signifikanter Rückgang der Ansiedelung von S. Enteritidis an den Tagen 7, 21 und 28 verzeichnet werden, außerdem konnte bei den Testgruppen kein Todesfall festgestellt werden, wohingegen die Sterblichkeitsrate der positiven Kontrollgruppe bei 15% lag. Ein signifikantes Absinken in der totalen Gesamtkeimzahl und der Gram-negativen Gesamtkeimzahl konnte für die Testgruppen im Vergleich zur positiven Kontrollgruppe an den Tagen 21 beziehungsweise 28 verzeichnet werden. Die Isolation von Bdellovibrionen aus den Testgruppen ergab keinen Unterschied zu den Kontrollgruppen. Die Lactobacillus-Bifidobacterium-Keimzahl war in den Testgruppen im Vergleich zur positiven Kontrollgruppe signifikant erhöht, vor allem am Tag sieben. Die C. perfringens-Keimzahl war in der positiven Kontrollgruppe gegenüber den Testgruppen am Tag 7 und 14 signifikant erhöht. Die Vergleiche des Körpergewichts der Testgruppen, negativer und überlebender Tiere der positiven Kontrollgruppe zeigten keinen signifikanten Unterschied. 5- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Stamm Nissle 1917 mit und ohne Plasmid auf die Reisolation von S. Enteritidis aus unterschiedlichen Teilen des Intestinaltraktes und aus Organen von SPF-Hühnern. Im Kropf, Proventrikel, Duodenum, Herzblut, Lunge, Leber und Milz konnte bei den Testgruppen im Vergleich zur positiven Kontrollgruppe eine Senkung der Reisolationsrate festgestellt werden, wohingegen sich keine Unterschied beim Vergleich der Caeca von positiver Kontrollgruppe und von den Testgruppen zeigte. 6- Untersucht wurden die Auswirkungen der oralen Applikation von E. coli Stamm Nissle 1917 mit und ohne Plasmid auf die Konzentrationen von Serum-Avidin, IgY gegen S. Enteritidis und IgY gegen E. coli Nissle 1917 bei experimentell mit S. Enteritidis-infizierten SPF-Hühnern. Bei der Serum-Avidin-Konzentration konnte kein Unterschied zwischen Testgruppen und positiver Kontrollgruppe festgestellt werden. Die Testgruppen zeigten ihrerseits im Vergleich zur positiven Kontrollgruppe eine signifikante Erhöhung der Konzentration von IgY gegen S. Enteritidis, vor allem an den Tagen 21 und 28. Ein signifikanter Anstieg der Konzentration von IgY gegen E. coli Nissle 1917 konnte in allen Testgruppen im Vergleich zur positiven Kontrollgruppe, vor allem an den Tagen sieben und vierzehn festgestellt werden. 7- Aus den Resultaten des ersten und des zweiten Versuchs schließen wir, dass der probiotische Stamm E. coli Nissle 1917 das Caecum von 7 bis 28 Tage alten Hühnern erfolgreich kolonialisieren kann. Orale Applikation des Stammes kann die Ansiedlung von S. Enteritidis, aerobe und Gram-negative Gesamtkeimzahl, Mortalitätsrate, Reisolationsrate von S. Enteritidis in Organen und im Intestinaltrakt (mit Ausnahme des Caecums im ersten Versuch) deutlich reduzieren. Die Kombination von E. coli Nissle 1917 und Bdellovibrio bacteriovorus hatte nicht den gewünschten Effekt, der auch von mehreren Autoren in-vitro beobachtet wurde. Bdellovibrionen waren in den Testgruppen des ersten Versuchs im Vergleich zu denen im zweiten deutlich erhöht. Beim durchschnittlichen Körpergewicht konnte kein signifikanter Unterschied zwischen beiden Versuchen festgestellt werden. Der Serum-Avidin-Spiegel war im ersten Versuch in der Testgruppe deutlich reduziert, im zweiten dagegen nicht. Dies ist möglicherweise auf den Effekt von Bdellovibrionen zurückzuführen, bedarf allerdings noch der weiteren Bestätigung. Im Allgemeinen konnten wir eine signifikante Verbesserung des Immunsystems der Vögel als Resultat der oralen Applikation von E. coli Nissle 1917 feststellen. 8- Untersucht wurden die antagonistischen Effekte von E. coli Stamm Nissle 1917 mit und ohne Plasmid auf die in-vitro-Wachstumsrate von S. Enteritidis PT4. Wir konnten feststellen, dass E. coli Nissle 1917 sowohl mit als auch ohne Plasmid die Wachstumsrate von S. Enteritidis erfolgreich reduzieren konnte. / Summary Influence of the probiotic strain Escherichia coli Nissle 1917 on experimental infections with Salmonella Enteritidis PT4 in chickens Imad Azmi Awadein Mohamed Institute of Bacteriology and Mycology, Faculty of Veterinary Medicine, University of Leipzig and Institute of Poultry Diseases, Faculty of Veterinary Medicine, Free University Berlin Submitted in January, 2004 Keywords: (E. coli strain Nissle 1917, chickens, S. Enteritidis, Probiotics, Bdellovibrios) (94 pages, 48 figures, 16 tables and 150 references) 1- The effect of oral application of E. coli strain Nissle 1917 with Bdellovibrio bacteriovorus on the colonization of S. Enteritidis PT4, reduction of the mortality rate, total aerobic bacterial counts, Gram negative bacterial counts, bdellovibrio bacterial counts and body weight of Specific Pathogen Free chickens were investigated. A significant reduction in the colonization of S. Enteritidis PT4 in the caecum of trial groups at 14, 21 and 28 days old in comparison to the positive control group was observed. A reduction in mortality rate in the trial groups was seen in comparison to the positive control group which had 10 %. The total aerobic and Gram negative bacterial counts were significantly decreased in the trial groups at 28 days old in comparison to the positive control group. Bdellovibrio bacterial counts were significantly increased in the trial groups in comparison to the control groups. No significant difference was seen in the mean body weight of the trial groups in comparison to control groups. 2- The effect of oral application of E. coli strain Nissle 1917 with Bdellovibrio bacteriovorus on reisolation rate of S. Enteritidis from different parts of the intestinal tract and organs of SPF chickens was investigated. A reduction in the S. Enteritidis reisolation rate was seen from crop, proventriculus, duodenum, heart blood, lungs, livers and spleens of the trial groups in comparison to the positive control group but not from the caecum. 3- The effect of oral application of E. coli strain Nissle 1917 and Bdellovibrio bacteriovorus on the serum avidin levels, IgY against S. Enteritidis and IgY against E. coli strain Nissle 1917 levels of SPF chickens experimentally infected with S. Enteritidis was investigated. A significant reduction in the serum avidin levels was observed in the trial groups at 7 days old in comparison to the positive control group. The antibody levels against S. Enteritidis was significantly increased in the trial group at 7 days old in comparison to the positive control group. A significant increase at day 7 in the antibody levels against E. coli strain Nissle 1917 was seen in the trial group in comparison to the positive control group. 4- The effect of oral application E. coli Nissle 1917 with and without plasmid on reduction on the colonization of S. Enteritidis PT4, reduction of the mortality rate, total aerobic bacterial counts, Gram negative bacterial counts, bdellovibrio bacterial counts, Lactobacillus-Bifidobacterium counts, C. perfringens bacterial counts and body weight of SPF chickens were investigated. A significant reduction in the colonization of S. Enteritidis PT4 in the caecum of trial groups at 7, 21 and 28 days old in comparison to the positive control group was observed. No mortalities were seen in the both trial groups in comparison to the positive control group which had 15 % mortalities. The total aerobic and Gram negative bacterial counts were significantly decreased in the trial groups at 21 and 28 days respectively old in comparison to the positive control group. No significant difference was observed in bdellovibrio isolation from the trial groups in comparison to the control group. Lactobacillus-Bifidobacterium bacterial counts were significantly increased in the trial groups at 7, 14 and 21 days old in comparison to the positive control group. A significant reduction in C. perfringens bacterial counts were seen in the trial groups at 7 and 14 days old in comparison to positive control group. No significant difference was observed in the mean body weight of the trial groups in comparison to control groups. 5- The effect of oral application E. coli Nissle 1917 with and without plasmid on the reisolation of S. Enteritidis from different parts of intestinal tract and organs of SPF chickens was investigated. A reduction in the S. Enteritidis reisolation rate was seen from crop, proventriculus, duodenum, caecum, heart blood, lungs, livers and spleens of the trial groups in comparison to the positive control group. 6- The effect of oral application E. coli Nissle 1917 with and without plasmid on the serum avidin levels, IgY against S. Enteritidis and IgY against E. coli strain Nissle 1917 levels of SPF chickens experimentally infected with S. Enteritidis were investigated. No significant reduction in the serum avidin levels were seen in the trial groups in comparison to the positive control group. The antibody levels against S. Enteritidis were significantly increased in the trial groups at 21 and 28 days old in comparison to the positive control group. A significant increase in the antibody levels against E. coli strain Nissle 1917 were seen in the trial groups in comparison to the positive control group. 7- From the results observed in the first and second experiments we concluded that probiotic strain E. coli Nissle 1917 can successfully colonize the caecum of SPF chickens from 7 to 28 days old. Oral appliction of probiotic strain E. coli Nissle 1917 has significantly reduced the S. Enteritidis colonization, total and Gram negative bacterial counts, mortality rate, reisolation rate of S. Enteritidis in organs and the intestinal tract except the caecum in first experiment in the trial groups in comparison to positive control. The combination between E. coli strain Nissle 1917 and B. bacteriovorus had not achieved the desirable effects in the first experiment which observed by many authors in vitro. Bdellovibrio was significantly increased in the trial groups of the first experiment in comparison to the second experiment. This may be a possible effect of Bdellovibrio which is yet to be confirmed. No significant difference was observed in the mean body weight in both experiment. A significant reduction of serum avidin levels were observed in the trial groups of the first experiment but not in the second experiment. We have observed a significant improvement in the immune system of the birds as a results of oral application of E. coli strain Nissle 1917. 8- Antagonistic effect of E. coli Nissle 1917 with and without plasmid on the growth rate of the S. Enteritidis PT4 in vitro was investigated. We concluded that E. coli Nissle 1917 with and without plasmid successfully reduced the growth rate of S. Enteritidis in comparison to the control one.

<p>Allergic diseases, for example asthma and eczema, are nowadays considered belonging to the most common chronic diseases amongst children in the West, but the cause for this increase in allergy prevalence is unknown. Since studies have indicated a connection between children's exposure of microorganisms during infancy and risk of developing allergic disease, it is suggested that this exposure is a crucial factor in question of allergy development or not. Other studies have established differences in microflora composition between healthy children and children with allergic disease, and several studies have shown that probiotic therapy can give positive results in both prevention and treatment of allergic diseases.</p><p>The aim of this master's thesis was to develop a method, using PCR and denaturing gradient gel electrophoresis, to study the diversity of <em>Lactobacillus spp</em>. in fecal samples retrieved from a study of the probiotic strain<em> L. reuteri</em> ATCC 55730. The developed method was successful in detecting lactobacilli in fecal samples, but three other bacterial genera commonly found in humans were also amplified. Comparison of average numbers of detected bacterial strains and lactobacilli strains between samples belonging to the probiotics and placebo groups, respectively, showed higher numbers for the probiotics group. Also, the only fecal samples that contained <em>L. reuteri</em> belonged to the probiotics group. Although the results are far from statistically significant, they support the theories that probiotics may influence the intestinal microbiota.</p>

Tez (Uzmanlık) - Süleyman Demirel Üniversitesi, Tıp Fakültesi, İç Hastalıkları Anabilim Dalı, 2008. / Kaynakça var.