The aim of this study was to evaluate quality parameters of dry fermented sausages. Samples of two types of dry fermented sausages were sampled – sausage “Poličan“ and “Lovecký salám“, from three different producers. Basic physical-chemical analysis of quality parameters were performed in sausage mixture and immediately after end of ripening (21 days for Poličan and 16 days for Lovecký salám). Results of quality parameters analysis were compared with requirements of Czech legislation and differences between producers were observed. There were significant differences between producers, however all of them met requirements set by 326/2001 Czech notification as amended of Ministry of agricultural.

Tiger nuts (Cyperus esculentus L) are recognized as a high potential, alternative source of food nutrients. However, there is limited scientific literature on the technological possibilities for developing value-added foods, such as fermented products from tiger nut milk. Therefore, strategies for producing and improving the properties of fermented tiger nut milk were investigated for generating lactose-free, nutritious yogurt-like products with acceptable sensory properties and a prolonged shelf life quality. A wet-milling procedure was standardized for extracting tiger nut milk from tiger nuts, and the effects of the extraction process on nutrient distribution, colour properties and colloidal stability of the milk were analyzed. Next, tiger nut milk was enriched with proteins and/or hydrocolloids and the impact of the additives on the physical properties of the milk were determined. Enriched tiger nut milk was fermented by using classical yogurt cultures and the obtained products were analyzed for the microbiological, physico-chemical and sensory characteristics. Additionally, effects of enriching tiger nut milk with microbial transglutaminase cross-linked proteins on the microbiological and physico-chemical properties were evaluated. Higher wet-milling intensity improved the nutrient composition, colloidal stability and colour of the milk. Enrichment of tiger nut milk with milk proteins and xanthan gum enhanced the viscosity and stability, and after fermentation, led to homogenous gel-like products with superior microbiological, physico-chemical and different sensory properties compared to the fermented plain tiger nut milk. Microbial transglutaminase cross-linked proteins improved the physical characteristics of the fermented product, especially during storage. This product would be relevant in many developing countries with high prevalence of lactose intolerance, limited access to nutritious food but show a high distribution of tiger nut vegetation.:1. Introduction and aim 1 2. Literature review 4 2.1 Tiger nut, origin, nutritional value and food use 4 2.2 Tiger nut milk, preparation and nutrient composition 7 2.3 Colloidal characteristics of tiger nut milk 9 2.4 Factors accounting for the dispersion stability of tiger nut milk 10 2.5 Enhancing tiger nut milk stability 12 2.6 Properties of fermented tiger nut milk 17 2.7 Microbial transglutaminase and properties of fermented tiger nut milk 18 3. Methodology 21 3.1 Extraction and characterisation of tiger nut milk 21 3.1.1 Sample collection and preparation 21 3.1.2 Tiger nut milk extraction 21 3.1.3 Nutrient analysis of tiger nuts 22 3.1.4 Analysis of tiger nut products 23 3.1.5 Particle size distribution 24 3.1.6 Colloidal stability 25 3.1.7 Colour measurement 25 3.2 Stabilisation of tiger nut milk dispersion 26 3.2.1 Tiger nut milk preparation 26 3.2.2 Preparation of tiger nut milk enrichments 26 3.2.3 Gravitational stability of enriched tiger nut milk 27 3.2.4 Accelerated gravitational stability of enriched tiger nut milk 28 3.2.5 Viscosity of TNM mixtures 29 3.3 Extraction and characterisation of globular tiger nut proteins 29 3.3.1 Protein extraction and fractionation 29 3.3.2 Molecular mass of globular tiger nut proteins 31 3.3.3 Denaturation temperature of globular tiger nut proteins 32 3.3.4 Isoelectric point of globular tiger nut protein 33 3.4 Properties of fermented tiger nut milk enriched with proteins 34 3.4.1 Materials and Reagents 34 3.4.2 Preparation of plain and enriched tiger nut milk 34 3.4.3 Fermentation of plain and enriched tiger nut milk 35 3.4.4 Viable counts of starter cultures in fermented tiger nut milk systems 36 3.4.5 Chemical analysis of unfermented and fermented tiger nut milk 36 3.4.6 Physical analysis of fermented tiger nut milk products 37 3.4.7 Sensory analysis of fermented tiger nut milk products 38 3.5 Microbial transglutaminase and fermented tiger nut milk property 38 3.5.1 Preparation of plain and enriched tiger nut milk 38 3.5.2 Fermentation of plain and enriched tiger nut milk 39 3.5.3 Analysis of the enzymatically cross-linked proteins 39 3.5.4 Viable counts 40 3.5.5 pH and titratable acidity 40 3.5.6 Syneresis and viscosity 41 3.5.7 Colour of fermented tiger nut products 41 3.6 Statistical analysis 41 4. Results and discussion 43 4.1 Extraction and characteristics of tiger nut milk 43 4.1.1 Material recovery, mass transfer and yield of tiger nut solids 43 4.1.2 Nutrient composition of tiger nut products 45 4.1.3 Physical properties of tiger nut milk 48 4.1.3.1 Particle size distribution of extracted tiger nut milk 48 4.1.3.2 Colloidal stability of tiger nut milk 49 4.1.3.3 Colour stability of tiger nut milk 51 4.2 Stabilisation of tiger nut milk 53 4.2.1 Effects of enrichments on the stability of tiger nut milk 53 4.2.2 Effects of pH and temperature on the stability of enriched TNM 56 4.2.3 Effects of enrichments on the rheology of tiger nut milk 58 4.3 Tiger nut protein extraction and characterisation 60 4.3.1 Protein extraction and fractionation 60 4.3.2 Molecular mass of tiger nut protein 62 4.3.3 Thermal denaturation of tiger nut protein 63 4.3.4 Isoelectric point of tiger nut proteins 66 4.4 Properties of fermented tiger nut milk enriched with proteins 67 4.4.1 Acidification and gel formation during fermentation 67 4.4.2 Microbiological properties of fermented enriched tiger nut milk 70 4.4.3 Physico-chemical properties of fermented enriched tiger nut milk 71 4.4.4 Sensory properties of fermented tiger nut milk products 76 4.5 Microbial transglutaminase and fermented tiger nut milk property 77 4.5.1 Effects on tiger nut milk fermentation 77 4.5.2 Microbiological properties during storage of fermented product 81 4.5.3 Physico-chemical properties during storage of fermented product 83 4.5.4 Effects on colour of fermented tiger nut product 86 5. Conclusions and outlook 88 Bibliography 90 List of figures 111 List of tables 115 List of Publications 116 Poster and presentations 116 / Erdmandeln (Cyperus esculentus L) haben ein hohes Potential als alternative Quelle Lebensmittelinhaltsstoffen. Allerdings gibt es nur in begrenztem Ausmaß Literatur über technologische Möglichkeiten zur Entwicklung von Mehrwert-Lebensmitteln wie fermentierter Erdmandelmilch. Daher wurden Strategien zur Herstellung und Verbesserung der Eigenschaften von fermentierter Erdmandelmilch zur Erzeugung laktosefreier joghurtähnlicher Produkte mit akzeptablen sensorischen Eigenschaften untersucht. Für die Extraktion der Erdmandelmilch wurde ein Nassmahlverfahren standardisiert und der Einfluss des Verfahrens auf die Nährstoffverteilung, die Farbeigenschaften und die kolloidale Stabilität der Milch analysiert. Als nächstes wurde Erdmandelmilch mit Proteinen und/oder Hydrokolloiden angereichert, und der Einfluss der Additive auf die physikalischen Eigenschaften des Extrakts bestimmt. Angereicherte Erdmandelmilch wurde mit klassischen Joghurtkulturen fermentiert, und die mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften der Produkte wurden untersucht. Zusätzlich wurden Effekte der Anreicherung von Erdmandelmilch mit enzymatisch vernetzten Proteinen auf die mikrobiologischen und physikalisch-chemischen Eigenschaften bewertet. Eine höhere Nassmahlintensität verbesserte die Nährstoffzusammensetzung, die kolloidale Stabilität und die Farbe der Milch. Die Anreicherung erhöhte die Viskosität und Stabilität und führte nach der Fermentation zu homogenen gelartigen Produkten mit verbesserten mikrobiologischen, physikalisch-chemischen und sensorischen Eigenschaften im Vergleich zur fermentierten Erdmandelmilch. Mikrobielle Transglutaminase-vernetzte Proteine verbesserten die physikalischen Eigenschaften des fermentierten Produkts, insbesondere während der Lagerung. Dieses Produkt wäre in vielen Entwicklungsländern mit hoher Prävalenz von Laktoseintoleranz und begrenztem Zugang zu nahrhaften Lebensmitteln als Alternative von Interesse.:1. Introduction and aim 1 2. Literature review 4 2.1 Tiger nut, origin, nutritional value and food use 4 2.2 Tiger nut milk, preparation and nutrient composition 7 2.3 Colloidal characteristics of tiger nut milk 9 2.4 Factors accounting for the dispersion stability of tiger nut milk 10 2.5 Enhancing tiger nut milk stability 12 2.6 Properties of fermented tiger nut milk 17 2.7 Microbial transglutaminase and properties of fermented tiger nut milk 18 3. Methodology 21 3.1 Extraction and characterisation of tiger nut milk 21 3.1.1 Sample collection and preparation 21 3.1.2 Tiger nut milk extraction 21 3.1.3 Nutrient analysis of tiger nuts 22 3.1.4 Analysis of tiger nut products 23 3.1.5 Particle size distribution 24 3.1.6 Colloidal stability 25 3.1.7 Colour measurement 25 3.2 Stabilisation of tiger nut milk dispersion 26 3.2.1 Tiger nut milk preparation 26 3.2.2 Preparation of tiger nut milk enrichments 26 3.2.3 Gravitational stability of enriched tiger nut milk 27 3.2.4 Accelerated gravitational stability of enriched tiger nut milk 28 3.2.5 Viscosity of TNM mixtures 29 3.3 Extraction and characterisation of globular tiger nut proteins 29 3.3.1 Protein extraction and fractionation 29 3.3.2 Molecular mass of globular tiger nut proteins 31 3.3.3 Denaturation temperature of globular tiger nut proteins 32 3.3.4 Isoelectric point of globular tiger nut protein 33 3.4 Properties of fermented tiger nut milk enriched with proteins 34 3.4.1 Materials and Reagents 34 3.4.2 Preparation of plain and enriched tiger nut milk 34 3.4.3 Fermentation of plain and enriched tiger nut milk 35 3.4.4 Viable counts of starter cultures in fermented tiger nut milk systems 36 3.4.5 Chemical analysis of unfermented and fermented tiger nut milk 36 3.4.6 Physical analysis of fermented tiger nut milk products 37 3.4.7 Sensory analysis of fermented tiger nut milk products 38 3.5 Microbial transglutaminase and fermented tiger nut milk property 38 3.5.1 Preparation of plain and enriched tiger nut milk 38 3.5.2 Fermentation of plain and enriched tiger nut milk 39 3.5.3 Analysis of the enzymatically cross-linked proteins 39 3.5.4 Viable counts 40 3.5.5 pH and titratable acidity 40 3.5.6 Syneresis and viscosity 41 3.5.7 Colour of fermented tiger nut products 41 3.6 Statistical analysis 41 4. Results and discussion 43 4.1 Extraction and characteristics of tiger nut milk 43 4.1.1 Material recovery, mass transfer and yield of tiger nut solids 43 4.1.2 Nutrient composition of tiger nut products 45 4.1.3 Physical properties of tiger nut milk 48 4.1.3.1 Particle size distribution of extracted tiger nut milk 48 4.1.3.2 Colloidal stability of tiger nut milk 49 4.1.3.3 Colour stability of tiger nut milk 51 4.2 Stabilisation of tiger nut milk 53 4.2.1 Effects of enrichments on the stability of tiger nut milk 53 4.2.2 Effects of pH and temperature on the stability of enriched TNM 56 4.2.3 Effects of enrichments on the rheology of tiger nut milk 58 4.3 Tiger nut protein extraction and characterisation 60 4.3.1 Protein extraction and fractionation 60 4.3.2 Molecular mass of tiger nut protein 62 4.3.3 Thermal denaturation of tiger nut protein 63 4.3.4 Isoelectric point of tiger nut proteins 66 4.4 Properties of fermented tiger nut milk enriched with proteins 67 4.4.1 Acidification and gel formation during fermentation 67 4.4.2 Microbiological properties of fermented enriched tiger nut milk 70 4.4.3 Physico-chemical properties of fermented enriched tiger nut milk 71 4.4.4 Sensory properties of fermented tiger nut milk products 76 4.5 Microbial transglutaminase and fermented tiger nut milk property 77 4.5.1 Effects on tiger nut milk fermentation 77 4.5.2 Microbiological properties during storage of fermented product 81 4.5.3 Physico-chemical properties during storage of fermented product 83 4.5.4 Effects on colour of fermented tiger nut product 86 5. Conclusions and outlook 88 Bibliography 90 List of figures 111 List of tables 115 List of Publications 116 Poster and presentations 116

A pilot scale nitrogen (N) removal system was constructed and operated for approximately 365 days and was designed to remove inorganic total ammonia nitrogen (TAN) from solids-separated dairy manure. An anaerobic fermenter, upstream of the N removal reactor, produced volatile fatty acids (VFAs), to be used as an electron donor to fuel denitrification, and TAN at a COD:N ratio of 18:1. However, sufficient amounts of non-VFA COD was produced by the fermenter to fuel the denitrification reaction at an average NO3- removal rate of 5.3 ± 2 mg/L NO₃--N. Total ammonia N was removed from the fermenter effluent in an N removal reactor where a series of aerobic and anoxic zones facilitated aerobic TAN oxidation and anoxic NO₃- and NO₂- reduction. The minimum dissolved oxygen (DO) concentration allowing for complete TAN removal was found to be 0.8 mg/L. However, TAN removal rates were less than predicted using default nitrifying kinetic parameters in BioWin®, a biological modeling simulator, which indicated the presence of a nitrification inhibitor in fermented dairy manure. Furthermore, an N balance during the aerobic zone indicated that simultaneous nitrification-denitrification (SND) was occurring in the aerobic zone of the N removal reactor and was most apparent at DO concentrations below 1.3 mg/L. A series of nitrite generation rate (NGR) experiments confirmed the presence of an inhibitor in fermented dairy manure. A model sensitivity analysis determined that the most sensitive ammonia oxidizing bacteria (AOB) kinetic parameters were the maximum specific growth rate, , and the substrate half saturation coefficient, . Nitrifying inhibition terms of competitive, non-competitive, mixed competitive, and un-competitive were applied to the growth rate equation in BioWin® but an accurate representation of the observed TAN removal rates in the pilot scale system could not be found by adjusting the kinetic parameters alone. Reducing the default BioWin® hydrolysis rate by approximately 50% produced a more accurate calibration for all inhibition terms tested indicating that the hydrolyization of organic N in dairy manure is less than typical municipal waste water. / Master of Science

La barrière intestinale est la plus grande surface de contact entre le milieu extérieur et le milieu intérieur. Outre ses fonctions d'absorption des nutriments, elle exerce un rôle important de défense contre les agents indésirables (toxines, bactéries) contenus dans la lumière intestinale. Une augmentation de la perméabilité intestinale a été observée chez les patients atteints du syndrome de l'intestin irritable (SII) ou des maladies inflammatoires chroniques de l'intestin (MICI). Cette hyperperméabilité intestinale est contemporaine d'une hypersensibilité viscérale à la distension de la paroi intestinale. Des travaux récents rapportent également une augmentation de l'activité protéolytique du contenu intestinal dans le cadre de ces deux pathologies. Les estrogènes, par leurs propriétés anti-inflammatoires et leur capacité à moduler la perméabilité intestinale par activation de leurs récepteurs (REs) peuvent contribuer à l'amélioration des symptômes associés à ces pathologies digestives. Une variété de traitements médicaux a été utilisée pour la prise en charge thérapeutique du SII et de MICI. Cependant, les patients questionnent les cliniciens sur des conseils diététiques susceptibles d'améliorer leur qualité de vie. Ainsi, l'objectif de ce travail était d'évaluer les effets et les mécanismes d'action impliqués, d'un traitement par du germe de soja fermenté (SG) sur l'hyperalgésie viscérale et l'hyperperméabilité intestinale dans des modèles animaux mimant le SII et les MICI afin de proposer des futures allégations santé à ce produit. Le rationnel de l'évaluation de cet ingrédient était basé sur sa composition intéressante, à savoir, sa teneur en composés à propriétés estrogéniques (isoflavones) et sa capacité à inhiber les protéases (BBI). Dans un premier temps, nous avons montré qu'un traitement oral de 15 jours par le SG diminue de façon significative l'hypersensibilité viscérale, l'hyperperméabilité intestinale ainsi que l'augmentation de l'activité protéolytique induites par un stress de contrainte chez le rat. La diminution de la perméabilité intestinale implique une surexpression de l'occludine, protéine des jonctions serrées. De même, le traitement par du SG réduit la densité des mastocytes au niveau du côlon. Tous les effets préventifs du SG sauf ceux sur l'activité protéolytique sont estrogéno-dépendants car bloqués par l'antagoniste des REs. Dans un second temps, nous avons montré qu'un traitement préventif par le SG pendant 15 jours présente des effets protecteurs vis-à-vis d'une inflammation intestinale induite par du TNBS. Le SG atténue la sévérité de l'inflammation, l'hyperperméabilité, l'hypersensibilité et l'augmentation de l'activité protéolytique induites par la colite. Les effets anti-inflammatoires du SG sont à la fois dépendants des phytoestrogènes et du contenu de l'ingrédient en BBI. En conclusion, ces données sont prometteuses pour une future utilisation du SG dans la gestion thérapeutique du SII et des MICI comme traitement adjuvant / The intestinal barrier is the largest area of contact between the external environment and internal environment. In addition to its function of nutrient absorption, the intestinal barrier plays a key role of defense against noxious agents (toxins, bacteria) contained in the intestinal lumen. An increase in intestinal permeability was observed in patients with irritable bowel syndrome (IBS) or inflammatory bowel disease (IBD). This intestinal hyperpermeability was often associated with visceral hypersensitivity to colorectal distension. Recent studies also report an increase in the proteolytic activity in patients with IBS or IBD. Estrogens, through their anti-inflammatory properties and their ability to modulate intestinal permeability by activating estrogen receptors (ERs), can play an important role in these digestive diseases. A variety of medical therapies have been used for treatment of IBS and IBD. However, patients question clinicians about dietary suggestions to improve their symptoms and quality of life. Thus, the aim of this study was to evaluate the effects and mechanisms of action involved of a treatment with fermented soy germ (SG) on visceral hyperalgesia, intestinal hyperpermeability in animal models mimicking the IBS and IBD. The evaluation of this ingredient was based on its interesting composition, i.e its content of isoflavones and a family of serine protease inhibitors known as BBI. Initially, we demonstrated that an oral treatment of 15 days by SG significantly reduces visceral hypersensitivity, intestinal hyperpermeability and increased proteolytic activity induced by acute stress in the rat. Decreased intestinal permeability is due to overexpression of occludin, a transmembrane tight junction protein. Similarly, treatment with SG reduces the density of colonic mast cells. All preventive effects of SG except those on the proteolytic activity are estrogen-dependent because blocked by the antagonist of ERs. In a second step, we demonstrated that a treatment for 15 days with SG induces protective effects against intestinal inflammation induced by TNBS. SG reduces the severity of colitis, decreases TNBS-induced hyperpermeability, hypersensitivity and increased proteolytic activity. The anti-inflammatory effects of SG are estrogen and/or BBI dependent. In conclusion, these data are promising for future use of the SG as adjuvant therapy in IBS and IBD management

O tratamento antimicrobiano pode destruir o equilíbrio da microbiota gastrintestinal, podendo induzir sintomas clínicos, principalmente a diarréia. A influência de Lactobacillus casei Shirota sobre a microbiota intestinal foi avaliada em um estudo prospectivo, randomizado, duplo-cego e controlado. Sessenta e três crianças hospitalizadas com idade de 2 a 14 anos, sob tratamento com antibióticos &#946;-lactâmicos, foram randomizadas para receber o leite fermentado por L. casei Shirota, 108-9 UFC/mL, ou o placebo, durante o tratamento antimicrobiano. As amostras de fezes foram colhidas antes da administração do leite fermentado, durante o tratamento antibiótico e uma semana após o término do tratamento com o antimicrobiano e a ingestão do leite fermentado. O número de L. casei Shirota aumentou significativamente (p<0,001) durante o período de ingestão do leite fermentado. Foi observado na microbiota do grupo que recebeu o placebo um aumento na contagem de Pseudomonas aeruginosa (p<0,05) e Clostridium sp (p<0,05), principalmente no último período da terapia antimicrobiana. A alteração da microbiota intestinal em decorrência do tratamento antibiótico foi constatada pela diminuição de acetato (p<0,05), butirato (p<0,05) e formato (p<0,05). Embora nenhuma criança deste estudo tenha apresentado diarréia, na avaliação geral, a microbiota daquelas que receberam o leite fermentado mostrou uma recuperação precoce da microbiota intestinal. Foi observado que a variação da contagem bacteriana realizada não foi significativa para as crianças do grupo que recebeu o leite fermentado, enquanto que no grupo placebo a contagem bacteriana ficou alterada, mostrando desequilíbrio da microbiota. Cerca de 50% das crianças ainda apresentaram L. casei Shirota nas fezes após uma semana da ingestão do leite fermentado. Este estudo mostrou que a ingestão do leite fermentado contendo L. casei Shirota promoveu um reequilíbrio mais rápido da microbiota intestinal quando comparada com a do grupo que ingeriu o placebo. / Antimicrobial treatment can destroy the balance of gastrointestinal microflora, which may induce clinical symptoms, mainly diarrhoea. The influence of Lactobacillus casei Shirota on the intestinal microflora was assessed in a prospective, randomised, double-blind controlled study. Sixty-three hospitalised children, with ages between 2 and 14 years, under treatment with &#946;-lactam antibiotics were randomised to receive milk fermented by L. casei Shirota, 108-9 CFU/mL, or placebo during the antimicrobial treatment. Stool samples were collected before the administration of fermented milk, during the antibiotic treatment, and one week after the end of treatment with the antimicrobial agent and the ingestion of fermented milk. The number of L. casei Shirota increased significantly (p<0.05) during the period in which fermented milk was ingested. An increase in the Pseudomonas aeruginosa (p<0.05) and Clostridium sp (p<0.05) count was observed in the microflora of the group that received placebo, mainly in the last period of antimicrobial therapy. The alteration of intestinal microflora as a result of antibiotic treatment was found by the reduction of acetate (p<0.05), butyrate (p<0.05) and formate (p<0.05). The variation in bacterial count proved not to be significant for the children under antimicrobial treatment who received fermented milk, while the placebo group showed imbalance of microflora with the result of the altered bacterial count. About 50% of the children still presented L. casei Shirota in their stools after interrupting the ingestion of fermented milk for one week. This study showed that ingestion of fermented milk containing L. casei Shirota promoted a much faster re-balance of the intestinal microflora when compared to the group that ingested a placebo.

A ensilagem de grãos de milho reidratados é uma estratégia de armazenamento vantajosa, economicamente viável e que melhora a digestibilidade do amido. Este estudo foi dividido em dois experimentos - Experimento 1: Utilização de aditivos químicos e microbianos sobre aspectos fermentativos e estabilidade aeróbia de silagens de milho reidratado. Os tratamentos consistiram em silagens de milho tratadas com Lactobacillus buchneri (LB - 5×105 ufc/g da matéria natural); Bactérias homoláticas (Homo - 5×105 ufc/g MN); Lactobacillus buchneri + homoláticas (Combo - 1×106 ufc/g MN); Lactobacillus buchneri + nitrito de sódio (LB - 5×105 ufc/g MN + 1,5 g/kg MN de nitrito) e benzoato de sódio (Benz - 2,0 g/kg MN). Foi utilizado o delineamento experimental inteiramente casualizado com 6 tratamentos e 4 repetições. Experimento 2: Utilização de silagem de grãos de milho reidratados com ou sem a adição de benzoato de sódio sobre a digestibilidade e desempenho de vacas leiteiras. Foram utilizadas 18 vacas Holandês, alocadas em três quadrados latinos para avaliação dos tratamentos: Controle (silagem de milho reidratado); Benz (silagem de milho reidratado + benzoato de sódio - 2,0 g/kg MN) e MiS (milho seco moído). Em todas as dietas o nível de inclusão do milho foi de 17,3%. No primeiro experimento, a composição química das silagens foi adequada e as perdas fermentativas foram baixas (<1,6%) para todos os tratamentos. A presença de aditivos microbianos resultou em silagens bem fermentadas, enquanto silagens Benz preservaram carboidratos solúveis e resultaram em menor concentração de produtos de fermentação. A estabilidade aeróbia foi máxima para LB, Combo e Benz (240h) e intermediária para LBNit (151h). A silagem inoculada com bactérias homoláticas foi menos efetiva em promover estabilidade aeróbia (74,7h), comparada à controle (53,5h). Os melhores parâmetros de degradabilidade ruminal da MS foram obtidos para a silagens LB e Combo. Silagem Benz e Homo foram semelhantes à Controle e o LBNIT piorou a degrabilidade ruminal da MS. No segundo experimento, O CMS foi similar entre os tratamentos, com média de 21,1 kg/dia, ao passo que a digestibilidade do amido e CNF foi maior para os grãos fermentados. O maior aporte energético das silagens gerou tendência de aumento na produção de leite corrigido para 3,5% de gordura. Ao relacionar essa produção com o CMS, os animais alimentados com silagens de grãos reidratados tenderam a uma maior eficiência alimentar. Vacas leiteiras alimentadas com silagens apresentaram menor excreção de nitrogênio ureico no leite (11,9 vs 13 mg/dL), o que sugere um melhor uso de nitrogênio para produção de proteína microbiana. O uso dos aditivos benzoato de sódio, L. buchneri e L. buchneri combinado com bactérias homoláticas é aconselhável para promover a fermentação, estabilidade aeróbia e valor nutritivo de silagens de grãos de milho reidratados. / Rehydrated corn grain silage is an advantageous storage method, economically viable and improves starch digestibility. This study was carried out in two experiments - Experiment 1: Use of chemical and microbial additives on fermentation aspects and aerobic stability of rehydrated corn grain silage. The treatments consisted of rehydrated corn silage with different types of additives: control (no additive); Lactobacillus buchneri (LB - 5 × 105 cfu/g of fresh matter); homolactic bacteria (Homo - 5 × 105 cfu/g FM); Lactobacillus buchneri + homolactic bacteria (Combo LB - 5 × 105 cfu/g + LP - 5 × 105 cfu/g FM); Lactobacillus buchneri + sodium nitrite (LB - 5 × 105 cfu/g FM + nitrite - 1.5 g/kg FM) and sodium benzoate (Benz - 2.0 g/kg FM). The experiment was made in a completely randomized design with six treatments and four replications. Experiment 2: Use of rehydrated corn silage with or without sodium benzoate on the digestibility and performance of dairy cows. Eighteen Holstein cows were distributed in a replicated 3×3 latin square design with three periods (22-d) to evaluate the effect of control treatment (rehydrated corn silage); Benz (rehydrated corn silage plus sodium benzoate - 2.0 g/kg FM) and DGC (dry ground corn). In all diets the inclusion of corn was 17.3% in dry matter basis. In the first experiment, the chemical composition of silages was adequate and the fermentative DM losses were low (<1.6%) for all treatments. The presence of microbial additives resulted in well fermented silages, while Benz silages preserved WSC and showed lower concentration of fermentation products. The aerobic stability was highest for LB, Combo and Benz (240h) and intermediate for LBNit (151h). The Homo silages was less effective in promoting aerobic stability (74.7h), compared to the control (53.5h). The best parameters of ruminal DM degradability were obtained for LB and Combo silages. Benz and Homo silages were similar to the control, while LBNIT worsened the ruminal degrability of DM. In the second experiment, dry matter intake was similar across treatments with average of 21.1 kg/day, whereas starch and CNF digestibility was higher for fermented grains. The higher energy of silages tended to increase the production of 3.5 % fat-corrected milk. Therefore, animals fed rehydrated grain silages tended to present increased feed efficiency. Dairy cows fed silages had lower excretion of milk urea nitrogen, suggesting a better use of nitrogen for microbial protein synthesis. The use of additives sodium benzoate, L. buchneri and L. buchneri combined with homolactic bacteria is advisible to improve the fermentation, the aerobic stability and the nutritive value of rehydrated corn grain silages.

This study aimed to evaluate the effect of vegetable by-products, including fruit by-products (passion fruit, orange, acerola, and mango) and soy by-product (okara), on the folate production by starter and probiotic strains for the bio-enrichment of fermented soy products. In the first part of this study, the impact of amaranth flour on folate production by these microorganisms was also evaluated. The effect of vegetable by-products and amaranth flour on the ability of three starters - Streptococcus thermophilus (ST-M6, TH-4, and TA-40) and ten probiotic strains (Lactobacillus (Lb.) acidophilus LA-5, Lb. fermentum PCC, Lb. reuteri RC-14, Lb. paracasei subsp. paracasei Lb. casei 431, Lb. paracasei subsp. paracasei F19, Lb. rhamnosus GR-1, and Lb. rhamnosus LGG, Bifidobacterium (B.) animalis subsp. lactis BB-12, B. longum subsp. longum BB-46, and B. longum subsp. infantis BB-02) to produce folate was evaluated, using a modified MRS broth. Most of the microorganisms were able to produce folate. However, folate production was strain-dependent and also dependent on the environmental conditions and on the vegetable substrate used. Passion fruit by-product presented the lowest folate concentration and was selected for the following experiments. Thus, the impact of the supplementation of soymilk with passion fruit by-product and/or commercial prebiotic fructooligosaccharides FOS P95 on the folate production by three St. thermophilus strains, as well as four probiotic Lactobacillus strains (LA-5, LGG, PCC, and RC-14) were evaluated. St. thermophilus ST-M6 and TH-4 produced the highest amounts of folate in all fermented soymilks. The concentration of the vitamin was also high when these strains grew in co-culture with LA-5 and LGG. Soymilk supplemented with both passion fruit by-product and FOS together presented the highest concentration of folate when fermented by the co-culture TH-4+LGG. This co-culture was selected to produce four fermented soy products (FSP). All FSP were bio-enriched with folate produced by the co-culture and the probiotic strain LGG remained always above 8 log CFU/mL until the end of the storage period (28 days at 4ºC). In contrast, the concentration of the vitamin was stable until day 14 then a slight decrease was observed at the end of the storage period. The FSP supplemented with both passion fruit by-product and FOS together may contribute with around 14% of the recommended daily intake for folate if consumed until day 14 of storage. During the in vitro simulated gastrointestinal conditions, the folate content of the digested FSP increased from 1.3 to 3.6-fold, especially at the small and large intestinal in vitro phases and the strain LGG was recovered. In contrast, St. thermophilus TH-4 was not recovered during the assay. Finally, the prebiotic potential of the bioactive compounds present in the fruit by-products was characterized. Fruit by-product water extracts (FWE) containing soluble fibres from fruit by-products were obtained through a hot-water extraction and were associated to phenolic compounds and showed antioxidant activity. The FWE (especially, orange and mango water extracts) presented an anti-inflammatory potential by decreasing the nitric oxide concentration produced in vitro by macrophages stimulated with lipopolisaccharides (LPS) from Salmonella Thyphymurium. The FWE (especially from mango) were able to stimulate the growth of the strains TH-4 and LGG, as well the folate production by these microorganisms when tested individually and in co-culture. The FWE also increased the adhesion of TH-4 and LGG to Caco-2 cells in an in vitro model. These results suggest a prebiotic potential of the fruit by-products evaluated and their potential towards increased folate production by the selected microorganisms. Therefore, the bio-enrichment of fermented soy products with folate produced by beneficial microorganisms is an alternative for the development of functional foods with high folate content. Additionally, fermentable bioactive compounds with functional and/or biological activity, such as soluble fibres associated to phenolic compounds with antioxidant activity, present in the fruit by-products, may act as potential prebiotic ingredients. These bioactive molecules may represent a potential natural alternative to synthetic drugs for the treatment of inflammatory processes. / O objetivo deste trabalho foi avaliar o efeito de subprodutos vegetais, incluindo subprodutos do processamento de fruta (maracujá, laranja, acerola e manga) e de soja (okara) na produção de folatos de novo por microrganismos strater e probióticos para bioenriquecer um produto de soja fermentado. Na primeira etapa deste trabalho, o impacto da farinha de amaranto na produção de folatos pelos microrganismos também foi avaliado. Neste sentido, primeiramente, verificou-se o efeito desses subprodutos vegetais e da farinha de amaranto na capacidade de três cepas starter - Streptococcus thermophilus (ST-M6, TH-4 e TA-40) e 10 cepas probióticas (Lactobacillus (Lb.) acidophilus LA-5, Lb. fermentum PCC, Lb. reuteri RC-14, Lb. paracasei subsp. paracasei Lb. casei 431, Lb. paracasei subsp. paracasei F19, Lb. rhamnosus GR-1, and Lb. rhamnosus LGG, Bifidobacterium (B.) animalis subsp. lactis BB-12, B. longum subsp. longum BB-46, e B. longum subsp. infantis BB-02) em produzir folato utilizando um caldo MRS modificado. A maior parte dos microrganismos testados foi capaz de produzir folato. Entretanto, a produção foi considerada cepa-dependente e, também, dependente das condições ambientais e do tipo de subproduto vegetal empregado. O subproduto de maracujá apresentou a menor concentração de folato e, por isso, foi selecionado para os testes seguintes. Neste sentido, o impacto da suplementação do leite de soja com subproduto de maracujá e/ou com o prebiótico comercial fruto-oligosacarídeo FOS P95 na produção de folato pelas três cepas de St. thermophilus, bem como quarto cepas probióticas do gênero Lactobacillus (LA-5, LGG, PCC e RC-14), também foi avaliado. Em cultura pura, as cepas de St. thermophilus ST-M6 e TH-4 produziram grande quantidade de folato nas formulações de extrato de soja fermentados. A concentração da vitamina foi maior quando tais cepas se desenvolveram em co-cultura com LA-5 e LGG. Observou-se que o extrato de soja suplementado concomitantemente com subproduto de maracujá e FOS apresentou a maior quantidade de folato quando fermentado pela co-cultura TH-4+LGG. Esta co-cultura, portanto, foi selecionada para desenvolver os produtos fermentados de soja (PFS). Todas as formulações foram bioenriquecidas e a cepa LGG manteve-se viável por todo o período de armazenamento (28 dias a 4ºC). Entretanto, a concentração da vitamina manteve-se estável apenas até o dia 14, observando-se uma diminuição da quantidade de folato ao final do período de armazenamento. Constatou-se que o produto fermentado de soja suplementado concomitantemente com subproduto de maracujá e FOS pode contribuir com cerca de 14% da ingestão diária recomendada para folato se consumido até o dia 14 do armazenamento. Além disso, durante a simulação gastrointestinal in vitro, observou-se que a digestão aumentou de 1,3 a 3,6 vezes a concentração da vitamina incrementando, consideravelmente, a bioacessibilidade do folato, principalmente nas porções simuladas do intestino delgado e grosso do intestino e a cepa LGG foi recuperada. Entretanto, a cepa St. thermophilus TH-4 não foi recuperada durante o ensaio. Por fim, o potencial prebiótico de componentes bioativos presentes nos subprodutos de fruta foi caracterizado. Uma extração Hot Water foi conduzida, a fim de obter extratos aquosos de subprodutos de fruta ricos em fibras solúveis associadas a compostos fenólicos com atividade antioxidante. Observou-se, ainda, que tais extratos aquosos de subprodutos de fruta (laranja e manga) apresentaram potencial anti-inflamatório constatado pela diminuição da concentração de óxido nítrico produzido por macrófagos estimulados com lipopolissacarideo (LPS) de Salmonella Typhymurium in vitro. Além disso, os extratos aquosos de subprodutos de fruta (principalmente o extrato aquoso de subproduto de manga) foram capazes de estimular a multiplicação das cepas TH-4 e LGG, bem como a produção de folatos por estes microrganismos quando avaliados individualmente e em co-cultura. Adicionalmente, esses extratos aquosos de subprodutos de fruta aumentaram a adesao do TH-4 e do LGG a células Caco-2 em modelo in vitro. Neste sentido, os resultados sugerem um potencial prebiótico dos subprodutos de fruta testados, de modo a estimular, não somente o desenvolvimento dos microrganismos avaliados mas, principalmente, o potencial destes em produzir folatos na presença dos substratos vegetais testados. O bioenriquecimento dos produtos fermentados de soja com folatos produzidos por microrganismos benéficos emerge como alternativa de alimento potencialmente funcional com alto teor de folato. Adicionalmente, compostos bioativos fermentescíveis e com atividade biológica como, por exemplo, as fibras solúveis associadas a compostos fenólicos com atividade antioxidante, presentes nos subprodutos de fruta testados podem constituir potenciais ingredientes prebióticos, além de representarem uma possível alternativa natural para o tratamento de processos inflamatórios.

Bifidobacterium spp. são microrganismos probióticos que podem ser incorporados em produtos alimentícios. Entretanto, para que seus efeitos benéficos à saúde humana ocorram, é necessário que o número de células viáveis na hora do consumo seja, no mínimo, 106UFC/g. As bifidobactérias são sensíveis à elevada acidez e, por isso, torna-se necessária a busca por métodos que possam proteger a integridade da célula, sendo um deles a microencapsulação. Em uma primeira etapa do trabalho, Bifidobacterium lactis foi encapsulado em micropartículas de alginato e alginato modificado (alginatoquitosana, alginato-quitosana-sureteric e alginato-quitosana-acryl-eze) e sua sobrevivência e liberação das micropartículas em fluidos simulados do trato gastrintestinal foram mensuradas utilizando-se soluções tampão com pH 1,5, 5,6 e 7,5, na presença e na ausência de pepsina (3g/L), pancreatina (1g/L) e bile (10g/L). A liberação de células das micropartículas teve uma relação direta com o pH do tampão. A microencapsulação aumentou a taxa de sobrevivência de B. lactis, em comparação com células não encapsuladas, em soluções tampão com pH 1,5 sem a presença de enzimas. Em suco gástrico simulado com enzimas digestivas, por outro lado, foi observado que a pepsina proporcionou um efeito protetor sobre as células de B. lactis, e nesse caso, as taxas de sobrevivência do microrganismo estavam diretamente relacionadas com o grau de injúria das células. Em uma segunda etapa do trabalho, leites fermentados com Streptococcus salivarius ssp. thermophilus e Lactobacillus delbrueckii ssp. bulgaricus foram enriquecidos com culturas de Bifidobacterium lactis submetidas a quatro tratamentos diferentes: desidratação em temperatura ambiente, liofilização/congelamento, encapsulação em alginatoquitosana e encapsulação em alginato-quitosana-acryl-eze. A população sobrevivente de B. lactis foi determinada semanalmente no leite fermentado e também após tratamento simulando condições do trato gastrintestinal. Os resultados indicaram que na ausência de pepsina, as populações de B. lactis foram reduzidas drasticamente após o contato com tampão pH 1,5, não sendo possível a detecção de células viáveis livres ou encapsuladas após 120 minutos de teste. A presença de pepsina influenciou positivamente a recuperação de células viáveis de B. lactis em todas as condições testadas, mas as culturas na forma desidratada apresentaram melhores resultados que as culturas microencapsuladas ou liofilizadas. No caso do leite fermentado contendo as células desidratadas, a população de B. lactis, após o tratamento em suco gástrico com enzimas, foi superior à detectada no produto antes desse tratamento. Conclui-se que a microencapsulação não foi eficiente para proteger B. lactis em leite fermentado contra injúrias causadas pelo trato gastrintestinal simulado. / Bifidobacterium spp. are microorganisms that can be added to foods. However, the benefits for the human health occur when the numbers of viable cells in the moment of the consumption is at least 106CFU/g. Bifidobacteria are acid sensitive, and methods to protect cell integrity, such as microencapsulation, are needed. In the first part of the present study, Bifidobacterium lactis was encapsulated in microparticles of alginate and modified alginate (alginate-chitosan, alginate-chitosan-sureteric and alginate-chitosan-acryl-eze) and the survival and release from microparticles in simulated gastrointestinal conditions were measured, using buffers (pH 1.5, 5.6 and 7.5), in the absence and presence of pepsin (3g/L), pancreatin (1g/L) and bile. The release from microparticles presented a direct relationship with pH. When the pH was 1.5 and no enzyme was present, encapsulation improved the survival of B. lactis, when compared to free cells. However, pepsin had a protective effect on B. lactis, and the survival rate was directly related to the cells injury degree. In the second part of the study, fermented milk samples containing Streptococcus salivarius ssp. thermophilus and Lactobacillus delbrueckii ssp. Bulgaricus were supplemented with B. lactis submitted to four different treatments: dehydration at room temperature, freeze drying, encapsulation in alginate-chitosan and encapsulation in alginate-chitosaacryl-eze. The number of viable B. lactis cells in the fermented milk was determined weekly and also after treatment with simulated gastrointestinal conditions. Results indicated that in the absence of pepsin, the number of viable cells decreased significantly after contact with buffers (pH 1.5), and no viable cell was detected after 120 minutes. Pepsin improved the recovery of viable cells in the assayed gastric conditions, being the dehydrated cultures more resistant than other cultures. In fermented milk containing the dehydrated cells, the number of viable cells increased after treatment with simulated gastrointestinal fluids. Microencapsulation was not an effective procedure to protect B. lactis in fermented milk against injury caused by the simulated gastrointestinal tract.

Le gari, une semoule de manioc (Manihot esculenta, Crantz) fermentée, gélifiée et déshydratée, représente le produit alimentaire le plus populaire de lAfrique de lOuest. Les bactéries lactiques constituent lun des plus importants groupes de micro-organismes impliqués dans létape de la fermentation du manioc, principalement en raison de leurs rôles établis dans le développement de la saveur et la préservation des aliments obtenus. Dans le but de fournir au consommateur un produit sain, de qualité organoleptique constante et acceptable, des efforts damélioration et de contrôle de la fermentation du manioc ont été entrepris. Seize souches de bactéries lactiques ayant des caractéristiques technologiques appropriées, ont été précédemment isolées et sélectionnées au cours de la fermentation du manioc en vue de leur utilisation comme culture starter pour la production de gari. Lobjectif de ce travail est de contribuer à un meilleur contrôle de la fermentation du manioc à travers la production dun starter de bactéries lactiques lyophilisées pour la production de gari. A lissue de la première partie du travail, les souches Lactobacillus plantarum VE36, G2/25, L. fermentum G2/10 et Weissella paramesenteroides LC11 ont été sélectionnées sur base de : (i) leur capacité à survivre après un test de déshydratation dans une solution de glycérol de concentration croissante suivi dun marquage par des marqueurs fluorescents, (ii) leur taux de survie après la lyophilisation, (iii) leur capacité dutilisation dune source de carbone après la lyophilisation, (iv) leur stabilité au cours du stockage sous forme lyophilisée. Nous avons cependant noté que la plupart des souches lyophilisées perdaient plus de 50% de leur viabilité après 60 jours de stockage à basse température. Dans la seconde partie du travail, nous avons essayé de comprendre lun des mécanismes à lorigine de la perte de viabilité au cours du stockage des souches lyophilisées : loxydation des lipides membranaires. Pour ce faire, limpact de la dégradation des acides gras polyinsaturés sur la survie cellulaire ou le pouvoir dacidification de la souche lyophilisée W. paramesenteroides LC11 a été évalué pendant 90 jours de stockage dans différentes conditions de stockage (différentes températures, présence ou absence dair et dhumidité). Un dosage des produits primaires de dégradation des acides gras polyinsaturés a aussi été effectué pour les échantillons stockés dans des conditions daération. A lissue de cette étude, une corrélation a été établie entre la perte de viabilité cellulaire et/ou du pouvoir dacidification et la dégradation des acides gras polyinsaturés. Dans loptique de confirmer ces résultats, leffet de composés protecteurs tels que les cryoprotecteurs et/ou des antioxydants (acide ascorbique et/ou butyle hydroxytoluene) sur la viabilité cellulaire, lintégrité membranaire et loxydation des lipides de la souche lyophilisée W. paramesenteroides LC11, ont été évalués pendant 90 jours de stockage dans des conditions daération et en présence dhumidité. Après le 90ième jour de stockage, les acides gras membranaires totaux ainsi que ceux de la fraction polaire ont également été analysés. La présence des cryoprotecteurs et du butyle hydroxytoluene avait considérablement réduit la perte de viabilité et dintégrité membranaire, ainsi que la dégradation des lipides de la fraction polaire. Une possible relation entre loxydation des acides gras polyinsaturés, plus précisément ceux de la fraction polaire des lipides, et la perte de viabilité cellulaire au cours du stockage des souches lyophilisées, a été confirmée. Des corrélations entre loxydation des acides gras polyinsaturés de la fraction polaire des lipides et la perte dintégrité cellulaire et, dautre part, entre la perte dintégrité cellulaire et la perte de viabilité cellulaire, ont été établies. Alors que les phénomènes doxydation des lipides se déroulaient au cours du temps, la perte de viabilité cellulaire survenait dès les premières heures du stockage. Les changements dans la perméabilité membranaire de la souche W. paramesenteroides LC11 ont ainsi été évalués 20h après la lyophilisation, à travers les déterminations du nombre de cellules viables et de la conductivité dune solution de réhydratation dont nous avons fait varier la température, la concentration en sels (NaCl), protons et composés protecteurs (glycérol, saccharose, maltodextrine, glutamate monosodique). Une augmentation de la température et de la concentration en sels (NaCl) ou en glutamate monosodique, ainsi quune baisse de la concentration en protons entraînaient une augmentation de la concentration en substances ioniques dissoutes et une baisse du nombre de cellules viables. Cependant, une augmentation de la concentration en glycérol, sucrose ou maltodextrine entraînait une diminution de la concentration en substances ioniques dissoutes et un maintien de la viabilité cellulaire. Ces résultats nous suggèrent quil pourrait se produire une rupture cellulaire durant la déshydratation et que cette situation pourrait influencer la stabilité de la souche lyophilisée dès les premières heures du stockage.

Enterococcus faecium è un componente fondamentale del microbiota di diversi alimenti fermentati quali formaggi e salumi e viene spesso isolato in alto numero in alimenti pronti al consumo. É inoltre largamente utilizzato come probiotico sia per l’uomo che per gli animali. Allo stesso tempo, però, questa specie batterica rappresenta una delle cause principali di infezioni nosocomiali quali endocarditi ed infezioni al tratto urinario. Studi recenti hanno dimostato che la specie E. faecium è costituita da due sub-popolazioni principali: la prima è denominate hospital associated (HA) clade “A” ed include la maggior parte dei ceppi responsabili di infezioni umane; la seconda è chiamata community associated (CA) clade “B”, e contiene principalmente ceppi commensali dell’uomo. Analisi più approfondite hanno rivelato un ulteriore suddivisione all’interno del clade A, nel sub-clade A1 (che raggruppa la maggioranza dei ceppi clinici) e nel sub-clade A2, associato agli animali e più sporadicamente ad infezioni umane. Nel 2012, EFSA ha redatto una linea guida per la valutazione della sicurezza di E. faecium usato come probiotico per gli animali, concludendo che i cepi appartenenti all’hospital-associated clade non devono essere utilizzati in nutrizione animale. Comunque, la distinzione tra le due sub-popolazioni è stata fatta utilizzando dati ottenuti prevalentemente da isolati umani e animali e solo un numero limitato di ceppi isolati dagli alimenti è stato considerato. Obiettivo di questa tesi di dottorato è stato quello di valutare la sicurezza di E. faecium negli alimenti fermentati, considerando ceppi isolati da formaggi artigianali e prodotti carnei e utilizzando sia tecniche di genomica che analisi fisiologiche. Nessuno dei ceppi alimentari studiati è risultato parte del clade A1, ma un ceppo isolato da un salame stagionato pronto al consumo ha rivelato diversi tratti tipici dei ceppi A1, tra cui particolari IS, transposase e geni di resistenza agli antibiotici. Questi risultati, così come altri dati, sottolineano la necessità di approfondire le conoscenze circa il ruolo dei ceppi di E. faecium isolati da alimenti come fattore di rischio per la salute umana. / Enterococcus faecium is commonly found in high numbers in ready to eat foods, being a member of the bacterial communities of a variety of fermented foods, including cheese and sausages, and is widely used as human and animal probiotic. However, this bacterial species is a leading cause of nosocomial infection, mainly endocarditis and urinary tract infections. Recent studies have demonstrated that E. faecium species consists of two very distinct clades: the hospital associated (HA) clade “A”, which includes most of the strains responsible for human infections, and the community associated (CA) clade “B”, that contains primarily human commensal isolates. Deeper analysis revealed a further split within clade A into sub-clade A1 (which groups the vast majority of clinical isolates), and sub-clade A2, associated with animals and sporadic human infections. In 2012, the European Food Safety Authority has issued a guideline for the safety assessment of E. faecium used as animal probiotics, concluding the strains belonging to the hospital-associated clade should not be used in animal nutrition. However, the differentiation of the two clades has been performed using data mainly deriving from human and animal isolates, and only a limited number of strains from the food chain were considered. Aim of this doctoral thesis was to assess the safety of E. faecium in fermented food, considering strains isolated from artisanal cheese and meat products, and using both whole genome-based techniques and physiological studies. None of the food isolates studied in this work belong to the epidemic clade A1, however a strain isolated from a ready to eat salami revealed several A1-specific traits, such as specific IS, transposases and antibiotic resistance genes. These results, as well as other data, underline the emergency of deeper understanding the role of E. faecium isolated from fermented foods as risk factor for human health.