A diversity of human-assisted approaches to rehabilitate and boost coral health have been suggested and investigated throughout the past years. Vitamins and trace-metal supplementation is a well-known strategy in human medicine and aquaculture, but vitamin addition is not currently actively tested for coral growth and recovery. These molecules are essential cofactors that have been correlated with coral thermal resistance and upregulated in corals treated with beneficial microorganisms (i.e., probiotics). To assess the effects of B12, B6 and zinc supplementation on coral health, we conducted a 2-month experiment in an open-closed-loop system mesocosm joined to a peristaltic pump continuously dosing the vitamins and/or zinc to individual 250 L tanks. Fragments of five different colonies of Acropora hemprichii were randomly distributed into the respective treatment tanks (B12, B6, zinc, multi-treatment and control). After 21 days, the corals were exposed to a pulse (1 day) of thermal stress, followed by three weeks of recovery. Substantial mortality (55%) in the control treatment was observed during the stress and recovery, with B12, B6, zinc and multi treatments exhibiting significantly less mortality (<20%). Coral health data combined with analysis of microbiome and metabolomic approaches suggest that both vitamins and zinc have a positive effect on coral health recovery. This study is the first to provide evidence that complex B-vitamins accompanied by zinc supplementation, can be a valuable tool for coral reef rehabilitation, and paves the way to further understanding specific mechanisms by which these nutrients promote coral health will be needed.

Controlling pathogenic presence in broilers has become a priority in the poultry industry to prevent economic losses due to disease and infection, as well as the possible contamination of chicken products. The use of antibiotics reduces the incidence of infections; however, their removal from production initiated the search for suitable alternatives. Probiotic ineed supplements have been widely evaluated as alternatives. Probiotic use has improved broiler performance, reduced pathogenic loads, and stimulated the immune system at later life stages. However, there is still a gap in protection during the first weeks after the chick hatches. The in ovo supplementation of probiotics has the potential of promoting early health benefits and protect the chick against pathogens after hatch. In the present study, the in ovo inoculation of different probiotic species was evaluated. It was determined that the inoculation of higher concentrations of E. faecium (107 cfu/50µL) into the egg improves growth performance and intestinal morphology compared to lower doses (105 and 106 cfu/50µL). It was also determined that not all B. subtilis serotypes are safe for in ovo inoculation, even if recognized as safe for use in feed, due to a high reduction in hatchability. However, certain B. subtilis are safe for in ovo inoculation and regulate the gut microflora through modulations in coliforms and aerobic bacteria after hatch. Lastly, the in ovo inoculation of different Lactobacillus strains does not affect hatchability or growth performance. However, different Lactobacillus species stimulated cytokine production even during the first week of hatch. The bursa of Fabricius morphology was modulated through an increase in follicular area, which could possibly induce higher antibody production against incoming pathogenic challenges. These results indicate that the in ovo inoculation of probiotic bacteria can induce earlier benefits to broiler health through early changes in gut microflora, as well as early stimulation in the immune system. The early protection provided through the in ovo inoculation of probiotics combined with the protection obtained through the administration of probiotics in feed could potentially result in overall healthier broilers and therefore improved performance.

No description available.

No description available.

Amphibian populations have undergone unprecedented declines in recent decades. Many of these declines are due to the spread of the cutaneous fungal pathogen Batrachochytrium dendrobatidis (Bd), which causes the disease chytridiomycosis. The Panamanian golden frog (Atelopus zeteki) has not been seen in the wild since Bd spread through western Panama. In response to initial declines, golden frogs were collected from wild populations and placed in captive colonies with the goal of future reintroductions. An understanding of this species' natural defense mechanisms against Bd is needed for reintroduction to be successful. Previous studies indicate that cutaneous bacteria are an important defense mechanism for some amphibians and applying antifungal bacteria to the skin of Bd-susceptible amphibians (probiotic therapy) can prevent chytridiomycosis. Therefore, the goals of my dissertation were to characterize the bacterial community of A. zeteki and determine if probiotic therapy could be used to prevent chytridiomycosis in this species. I initially characterized the bacterial community of wild and captive golden frogs using samples collected prior to the initial declines and after approximately eight years in captivity. I found that the community structure of the microbiota was significantly different between wild and captive frogs; however, the offspring of the original captive frogs still shared 70% of their microbial community with wild frogs. Then, I characterized the Bd-inhibitory properties of 484 bacteria isolated from 11 species of free-living Panamanian amphibians. I found a large proportion of bacteria (75.2%) had the ability to inhibit Bd and this trait was widely distributed among bacterial taxa, although there was also significant variation within bacterial genera in their ability to inhibit Bd growth. I then experimentally tested the ability of four of these isolates to prevent chytridiomycosis in captive golden frogs. None of them successfully prevented infection; however, there were several frogs that cleared infection and this was correlated with composition of the bacteria initially present on their skin. Overall these results demonstrate that the structure of microbial communities of A. zeteki are important to host health and building on this might provide the best hope for reintroducing this iconic species back to its native habitat. / Ph. D.

Tesis por compendio / [EN] Celiac disease (CD) is a chronic enteropathy triggered by cereal gluten proteins in genetically predisposed individuals. The etiology is strongly associated with the genes of the human leukocyte antigen (HLA) encoding the DQ2/DQ8 molecules. Most CD patients carry this genotype but this is also present in the 40% of the general population and only a small percentage develops the disease. Thus, the HLA-DQ genotype is necessary but not solely responsible for the disease development. Gluten is the main environmental trigger but its intake neither fully explains the onset nor its clinical manifestations. Other environmental factors (e.g. early feeding practices, infections, intestinal microbiota) have been associated with the risk of developing CD. The only treatment for CD patients is the adherence to a gluten free diet (GFD), but the compliance with this dietary strategy is complicated because gluten is present in many foods. Therefore, the identification of modifiable environmental factors that contribute to CD onset is critical for the development of strategies to reduce its incidence. Observational studies in CD patients revealed imbalances in the intestinal microbiota which could contribute to the pathogenesis of the disease. It has been proposed that these imbalances are not only a secondary consequence of the disease but could also be a predisposing factor. To understand whether gut microbiota imbalances play a role in CD onset and pathogenesis, in vitro, animal and human prospective and intervention studies have been conducted in the context of the present PhD Thesis. The global aim of this Thesis has been to improve the understanding of the role played by intestinal microbiota in the pathogenesis and risk of CD, and the possibilities of contributing to disease prevention and treatment by modulating gut microbiota composition. Chapter 1 includes two in vitro studies investigating the influence of components of the gut microbiota (bifidobacteria and enterobacteria) on the maturation and functions of immunocompetent cells (dendritic cells), and on gluten toxicity in the intestinal epithelium (Caco-2 cells). We have observed that some Bifidobacterium strains are able to reduce the activation of dendritic cells and ameliorate the toxicity of gluten on intestinal epithelial cells. In Chapter 2 the effects of the administration of Bifidobacterium longum CECT 7347 was evaluated in an in vivo model of gluten induced enteropathy in newborn rats, resulting in a reduced proinflammatory cytokine production in the small intestine and CD4+T cell numbers in peripheral blood. Chapter 3 includes two observational studies in humans to unravel whether breast-feeding and human milk composition and/or the host genotype (HLA-DQ) are related to the microbiota, thereby influencing the later development of CD. We concluded that both factors may contribute to the early intestinal colonization of the infant's gut, influencing the Bifidobacterium spp. numbers. Human milk composition also varies in CD and non-CD mothers, modifying the supply of bifidobacteria and protective immune factors to the offspring. Finally, in Chapter 4 we have studied the potential beneficial effects of the administration of B. longum CECT 7347 in addition to the GFD to children with newly diagnosed CD. This study demonstrates that the bifidobacteria slightly reduces serum inflammatory markers and restored the gut microbiota composition. / [ES] La enfermedad celíaca (EC) es una enteropatía crónica de carácter autoinmune que sufren individuos genéticamente predispuestos tras la ingesta de gluten. La etiología está asociada con los genes del sistema "Antígeno Leucocitario Humano" (HLA) que codifican las moléculas DQ2/DQ8. Los pacientes con EC presentan este genotipo, sin embargo este también está presente en ¿40% de la población general y sólo un pequeño porcentaje desarrolla la enfermedad. Por lo tanto, el genotipo HLA-DQ resulta necesario pero no suficiente para que se desarrolle la enfermedad. El gluten es el principal desencadenante pero su ingesta tampoco explica su desarrollo ni sus manifestaciones clínicas. Otros factores ambientales (p.e. la lactancia, infecciones, microbiota intestinal) se han asociado con el riesgo de desarrollar la EC. El único tratamiento para los pacientes celíacos es el seguimiento de una dieta exenta de gluten (DEG), sin embargo su cumplimiento es complicado debido a que el gluten está presente en la mayoría de los alimentos procesados. Por ello, la identificación de factores ambientales modificables que contribuyan al desarrollo de la enfermedad, resulta fundamental para desarrollar estrategias que permitan reducir su incidencia. Estudios observacionales realizados en pacientes con la EC, han demostrado la existencia de desequilibrios en su microbiota intestinal, que podrían contribuir a la patogénesis de la enfermedad. Se ha propuesto que estos desequilibrios no son sólo una consecuencia secundaria de la EC, sino que podrían ser un factor predisponente. Para entender si la microbiota está implicada en el desarrollo y patogénesis de la EC, en la presente Tesis se han desarrollado estudios in vitro, con animales y estudios prospectivos y de intervención en humanos. El objetivo de esta Tesis ha sido avanzar en el conocimiento de la función que la microbiota intestinal desempeña en la patogénesis de la EC, así como, acerca de las posibilidades de tratar o prevenir esta enfermedad mediante la modulación de la composición de la microbiota intestinal. El Capítulo 1 incluye dos estudios in vitro en los que se ha estudiado la influencia de componentes de la microbiota intestinal (bifidobacterias y enterobacterias) en la maduración y las funciones de células inmunocompetentes (células dendríticas), y en la toxicidad del gluten en el epitelio intestinal (células Caco-2). Hemos observado que algunas cepas de Bifidobacterium son capaces de reducir la activación de las células dendríticas y de reducir la toxicidad del gluten sobre el epitelio intestinal. En Capítulo 2 incluye el estudio de los efectos de la administración de B. longum CECT 7347 en un modelo de enteropatía inducida por gluten en ratas recién nacidas, observándose una reducción de citoquinas proinflamatorias en el intestino y de células T CD4+ en sangre periférica. El Capítulo 3 incluye dos estudios observaciones en humanos en los que se ha investigado si la lactancia y composición de la leche materna y/o el genotipo (HLA-DQ) están relacionados con la microbiota y, si así, podrían influir en el desarrollo de la EC. Concluimos que ambos factores contribuyen a la colonización intestinal del niño en los primeros meses de vida, afectando especialmente al número de Bifidobacterium spp. La composición de la leche maternal varía entre madres celíacas y sanas, lo que podría modificar el aporte de bifidobacterias y factores inmunológicos protectores al lactante. Por último, en el Capítulo 4 incluye el estudio del potencial efecto beneficioso de la administración de B. longum CECT 7347 junto con la DEG en niños recién diagnosticados de EC. Este estudio demuestra que la bifidobacteria ligeramente reduce los marcadores inflamatorios en sangre periférica y contribuye a restablecer la composición de la microbiota intestinal. / [CA] La malaltia celíaca (MC) és una enteropatia crònica provocada per les proteines del gluten de cereals en individus predisposats genèticament. L'etiologia està fortament associat amb els gens de l'antigen leucocitari humà (HLA). Pacients amb MC porten aquest genotip però també està present en aproximadament el 40% de la població general i només un petit percentatge (1-3%) es desenvolupa la malaltia. Per tant, HLA-DQ genotip és necessària, però no l'únic responsable. El gluten és el principal desencadenant ambiental però la seva ingesta no explica completament l'inici ni les seves manifestacions clíniques. Altres factors ambientals, com la microbiota intestinal, s'han associat amb el risc de desenvolupar CD. Els estudis observacionals en pacients amb MC van revelar desequilibris en la microbiota intestinal que podria contribuir a la patogènesi de la malaltia. S'ha proposat que aquests desequilibris no només són una conseqüència secundària de la malaltia, però també podria ser un factor predisponent. Per entendre si els desequilibris microbiota intestinal podrien tenir un paper en l'aparició de CD, un estudi prospectiu amb el nen en risc la família està en marxa. L'únic tractament per als pacients amb EC és l'adherència a una dieta lliure de gluten, però el seu compliment és complicada a causa del gluten està present en molts aliments. La identificació de factors ambientals modificables que contribueixen a l'aparició de CD és fonamental per a les estratègies de desenvolupament que porten a una reducció de la incidència. Aquest pot ser el cas per als components de la microbiota intestinal, l'adquisició podria ser modulada per factors ambientals i dietètics. L'objectiu global de la tesi és desentranyar els coneixements actuals sobre el paper exercit per la microbiota intestinal en la patogènesi de l'EC, i les possibilitats de contribuir a la prevenció i tractament de la malaltia mitjançant la modulació de la composició de la microbiota intestinal En el Capítol 1 hem estudiat l'ús de models in vitro de la influència de la microbiota intestinal durant la maduració i funcions del sistema immunològic (cèl·lules dendrítiques), i les interaccions entre el gluten i intestinal intestí i la resposta de l'epiteli intestinal resultant d'aquesta interacció. Hem observat que alguns ceps de Bifidobacterium són capaços de reduir l'activació del sistema immune i millorar la resposta nociva de l'epiteli intestinal a l'estimulació amb gluten. En el Capítol 2 hem estudiat els efectes de l'administració d'una soca de Bifidobacterium (B. longum CECT 7347) per a un model animal de rates nounades. El tractament amb els bacteris es va associar amb una reducció en la producció de citocines proinflamatòries i la resposta immune de cèl·lules T CD4 +. En el Capítol 3 es va descriure que alguns genètic (HLA-DQ genotip) i factors ambientals (llet materna) influeixen en la colonització intestinal primerenca, especialment en Bifidobacterium spp., que poden influir en l'aparició de CD més tard. Finalment, en el Capítol 4 s'ha estudiat l'efecte probiòtic de l'administració de B. longum CECT 7347 en nens acabats diagnosticats d'EC i el seu paper en el restabliment de la salut intestinal. / Olivares Sevilla, M. (2015). Influence of intestinal microbiota in celiac disease pathogenesis and risk [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/58768 / Premios Extraordinarios de tesis doctorales / Compendio

Thesis (MSc (Agric)(Animal Science: Animal Nutrition)) -- University of Pretoria, 2007. / Includes bibliographical references.

No description available.

Lactobacillus is a genus of lactic acid bacteria frequently used as healthpromoting probiotics. Using probiotics to treat or prevent infections is a novel experimental approach with vast impact on future therapy. Lactobacillus rhamnosus GR-1 is a probiotic investigated for its ability to reduce urogenital disease including urinary tract infections caused by pathogenic Escherichia coli. L. rhamnosus GR-1 has been shown to modulate immunity, thought to influence its probiotic effect. In this thesis, the aim was to study immunomodulation by L. rhamnosus GR-1 and other lactobacilli, with emphasis on elicited immune responses such as nuclear factor-kappaB (NF-κB) activation and cytokine release from human urothelial cells. Viable, heat-killed, and isolated released products from L. rhamnosus GR-1 augmented NF-κB activation in E. coli-challenged urothelial cells. Blocking of lipopolysaccharide binding to toll-like receptor 4 completely quelled this augmentation. Size-fractionation, urothelial cell challenge, and two-dimensional gel electrophoresis of L. rhamnosus GR-1 released products presented several candidate proteins with NF-κB modulatory actions including chaperonin GroEL, elongation factur Tu, and a protein from the NLP/P60 protein family. While tumor necrosis factor was correspondingly augmented by L. rhamnosus GR-1, the release of two other cytokines, interleukin (IL)-6 and CXCL8, was reduced. Similar effects were observed in macrophage-like cells stimulated with L. rhamnosus GR-1. Many immunomodulatory effects of lactobacilli are believed to be species and strain dependent. Therefore, twelve Lactobacillus strains were used to screen for their effects on CXCL8 release from urothelial cells. A majority of these strains were able to influence CXCL8 release from the cells. Phylogenetic analysis revealed close evolutionary linkage between lactobacilli with similar actions on CXCL8. Increased knowledge on probiotic bacterial products and the mechanism(s) of action could lead to improved future treatments for infections.

Thesis (MSc (Microbiology))--Stellenbosch University, 2008. / Lactobacillus plantarum JW3BZ and Lactobacillus fermentum JW15BZ isolated from boza, a Bulgarian cereal based fermented beverage, produce bacteriocins JW3BZ and JW15BZ active against a wide range of food spoilage and pathogenic bacteria. Strains JW3BZ and JW15BZ are resistant to low pH (pH 2.0–4.0). Both strains grow well in MRS broth with an initial pH ranging from 5.0 to 10.0. Strain JW3BZ displayed intrinsic resistance to bile salts. Strain JW15BZ, on the other hand, is sensitive to bile salts exceeding concentrations of 0.3% (w/v). Both strains are weakly hydrophobic and are resistant to a wide range of antibiotics, antiinflammatory drugs and painkillers. Strains JW3BZ and JW15BZ adhered at 4% to Caco-2 cells and they did not compete with Listeria monocytogenes Scott A for adhesion. A homologue of MapA, a gene known to play a role in adhesion, was detected in L. plantarum JW3BZ. Both strains have high auto- and co-aggregation properties. Bacteriocin JW15BZ was partially purified with ammonium sulfate, followed by separation on Sep-Pak C18 and reverse phase High Pressure Liquid Chromatography (HPLC). Two separate peaks with antimicrobial activity were recorded for bacteriocin JW15BZ, suggesting that it consists of at least two antimicrobial peptides. Lactobacillus plantarum JW3BZ contains genes homologous to plnE, plnF and plnI of the plnEFI operon that encode for two small cationic bacteriocin-like peptides with double-glycine-type leader peptides and its respective immunity proteins. The antimicrobial activity displayed by strain JW3BZ may thus be ascribed to the production of plantaricins E and F. Bacteriocin JW3BZ and JW15BZ displayed activity against herpes simplex virus (HSV-1) (EC50=200 μg/ml). Both strains were identified in boza after 7 days at storage at 4 oC and repressed the growth of Lactobacillus sakei DSM 20017, indicating that the bacteriocins are produced in situ. The sensory attributes of boza prepared with different starter cultures did not vary considerably, although statistical differences were observed for acidity and yeasty aroma. Encapsulation of strain JW3BZ and JW15BZ in 2% sodium alginate protected the cells from low pH (1.6) and 2.0% (w/v) bile. The rate at which cells were released from the matrix varied, depending on the conditions. Better survival of strains JW3BZ and JW15BZ encapsulated in 2% (w/v) alginate was observed during 9 h in a gastro-intestinal model. Highest release of cells was observed at conditions simulating colonic pH (pH 7.4), starting from 56-65% during the first 30 min, followed by 87%. Complete (100%) release was recorded after 2.5 h at these conditions. Strains JW3BZ and JW15BZ could be used as starter cultures in boza. The broad spectrum of antimicrobial activity of bacteriocins JW3BZ and JW15BZ is an added advantage, rendering the cells additional probiotic properties. Encapsulation of the cells in alginate gel increased their resistance to harsh environmental conditions and may be the ideal method to deliver viable cells in vivo.