Thesis (PhD)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: Background: Irritable bowel syndrome (IBS) is a common gastrointestinal (GI) disorder. GI symptoms and impaired quality of life affect between 10-20% of all adults, corresponding to about 25-50% of all patients who visit a gastroenterologist’s clinic. In recent years, several novel mechanisms of IBS that likely relate to previously established theories have been identified. Inflammation, postinfectious low-grade inflammation, immunological and genetic predisposition along with altered microbiota are critical in IBS development, while several dietary factors may also play a role in this syndrome. However, none of these factors accounts for the full repertoire of IBS symptoms, and the pathophysiology of this condition is not fully understood. The overarching aim of this study was to investigate the nutrient intakes, GI microbiota and the effect of Lactobacillus plantarum (L.plantarum) 299v in IBS patients. Sub-aims: 1) Update healthcare professionals on current probiotic information and provide an overview of probiotic treatment approaches, with special emphasis on IBS, 2) conduct a well designed randomised, double blind, placebo-controlled trial (RCT) with L. plantarum 299v as part of an intervention and establish whether a course of probiotics may alleviate undesirable symptoms of IBS and improve quality of life, 3) assess nutrient intake in patients with irritable bowel syndrome (IBS) compared to dietary recommendations, 4) validate and assess the reproducibility of food records and 5) identify possible nutrient risk components for establishing GI microbiota involved in IBS and as part of an intervention, determine whether a course of probiotics may alter stool microbiota. Results: 1) A review article published by the author provides an overview of current probiotic treatment options to health care professionals and indicates certain probiotics are a promising therapeutic treatment option for management of IBS symtpoms, 2) the effects of the single strain probiotic, L. plantarum 299v, supplementation was evaluated in a RCT. Compared to placebo, the probiotic supplementation showed no significant reduction in GI symptom severity scores, particularly abdominal pain relief. Quality of life was also not improved in the treatment versus control group. Both the treatment and placebo groups improved significantly over the trial period, indicating a large placebo effect, 3) nutrient intakes of the IBS patients compared to current dietary reference recommendations indicates that this group of patients are at risk for nutrient inadequacies in key macro and micronutrients, 4) the validity and reliability of the dietary data showed good reliability but poor validity as measured by plasma fatty acids and 5) the GI microbiota composition in the phenotypically different diarrhoea-predominant IBS (D-IBS) vs. constipation-predominant IBS (C-IBS) showed that D-IBS patients had significantly lower counts of Lactobacillus plantarum compared to C-IBS patients. The probiotic had no significant effects on the GI microbiota as measured by quantitative polymerase chain reaction (qPCR). It was found that nutrient intakes had a significant impact on the microbiota. Lower fibre intakes were associated with higher Bacteroides spp., lower Bifidobacteria bifidum and Lactobacillus plantarum counts in both IBS groups. Conclusion: Taken together, L.plantarum 299v did not alleviate the GI symptoms of IBS, nor was it associated with significant changes in the GI microbiota. IBS patients may be at risk of key nutrient inadequacies. The influence of nutrient intakes on the GI microbiota provides an attractive explanation as a potential pathophysiological factor for IBS. / AFRIKAANSE OPSOMMING: Agtergrond: Prikkelbare derm-sindroom (PDS) is ‘n algemene gastro-intestinale (GI) stoornis. GI simptome affekteer die lewenskwaliteit van 10-20% van alle volwassenes. Dit stem ooreen met ongeveer 25-50% van alle pasiënte wat ‘n gastroënteroloog konsulteer. Verskeie oorspronklike meganismes vir die ontwikkeling van PDS is onlangs identifiseer. Inflammasie, post-infektiewe lae-graadse inflammasie, immunologiese en genetiese vatbaarheid tesame met veranderde mikrobiota is krities vir die ontwikkeling van PDS. Sekere dieetfaktore mag ook bydraend wees tot hierdie sindroom. Geen van hierdie faktore is egter verantwoordelik vir die volle spektrum van PDS simptome nie en die patofisiologie van die toestand word ook nog nie ten volle verstaan nie. Die oorkoepelende doel van hierdie studie is om nutriëntinname, GI mikrobiota en die uitwerking van L.plantarum 299v in PDS pasiënte bepaal. Sub-doelwitte: 1) Om gesondheidswerkers in te lig aangaande die nuutste inligting oor probiotika en om ‘n oorsig van probiotika behandelingsopsies te verskaf, met spesiale klem op PDS, 2) om ‘n goed beplande ewekansige, dubbel-blinde, plasebo-beheerde kliniese studie met L.plantarum 299v as deel van die intervensie uit te voer om sodoende te bepaal of ‘n kursus probiotika ongewensde simptome van PDS kan verbeter en lewenskwaliteit sodoende verhoog, 3) om nutriëntinname in pasiënte met PDS te bepaal vergeleke met dieet aanbevelings, 4) om die geldigheid en herhaalbaarheid van voedselrekords te bepaal en 5) om moontlike nutriënt risikokomponente vir die ontwikkeling van GI mikrobiota betrokke in PDS te identifiseer en om as deel van ‘n intervensie te bepaal of ‘n kursus probiotika stoelgang mikrobiota patrone verander. Resultate: 1) ‘n Oorsigartikel gepubliseer deur die kandidaat dui probiotika aan as ‘n belowende terapeutiese opsie in die behandeling van PDS simptome, 2) die effek van ‘n enkelstam probiotikum, L.plantarum 299v, is evalueer deur ‘n ewekansige, dubbel-blinde, plasebo-beheerde kliniese studie. Vergeleke met die plasebo, het probiotiese aanvulling geen betekenisvolle vermindering in die GI simptome in PDS pasiënte tot gevolg gehad nie. Lewenskwaliteit het ook nie verbeter in die behandelde versus die kontrole groep nie. Beide die behandelde en plasebo groepe het aansienlik verbeter oor die studietydperk, wat ‘n groot plasebo effek aandui, 3) nutriëntinname van die PDS groep vergeleke met huidige dieetaanbevelings, dui daarop dat hierdie groep pasiënte ‘n risiko het vir die ontwikkeling van kern nutriënttekorte (makro- en mikronutriënte), 4) die geldigheid en betroubaarheid van die dieetdata dui op goeie betroubaarheid, maar swak geldigheid soos bepaal deur plasma vetsure en 5) die dermkanaal mikrobiotiese samestelling in die verskillende fenotipes, diarree-oorheersende PDS (D-PDS) vs. konstipasie-oorheersende PDS (K-PDS) dui daarop dat D-PDS pasiënte aansienlike minder Lactobacillus plantarum gehad het vergeleke met K-PDS pasiënte. Die probiotikum het geen beduidende uitwerking op die oorheersende mikrobiota gehad nie, soos gemeet deur kwantitatiewe polimerase kettingreaksie (kPKR). Daar is gevind dat dieet ‘n beduidende impak op die mikrobiota gehad het. Daar is ‘n verband tussen laer vesel inname en hoёr Bacteroides spp. en laer Bifidobacteria bididum en Lactobacillus plantarum tellings gevind in beide PDS groepe. Gevolgtrekking: Die L.plantarum 299v enkelstam probiotikum het nie die gastrointestinale simptome van PDS pasiënte verlig nie en daar is ook geen beduidende veranderinge in die mikrobiota gevind nie. PDS pasiënte mag ‘n verhoogde risiko toon vir kern nutriënttekorte. Die invloed van nutriëntinname op GI mikrobiota verskaf ‘n belowende verduideliking as ‘n potensiële patofisiologiese faktor in PDS.

Bakgrund: IBS (Irritabel Bowel Syndrome) är en funktionell mag-tarmsjukdom, med en oklar patofysiologi och etiologi. IBS är en vanlig åkomma hos den västerländska befolkningen och karakteriseras av återkommande buksmärta/obehag, uppblåsthet, diarré och/eller förstoppning samt ökad gasbildning. Det finns idag inget botemedel för IBS, men intresset för probiotika som behandling vid IBS har på senare tid ökat. Probiotika, vilket definieras som levande mikroorganismer, som när de intas i adekvata mängder ger en hälsovinst hos värden tros ha en symptomatisk effekt vid IBS, och har därmed varit av intresse för behandling av IBS. Syfte: Syftet med denna litteraturstudie är att undersöka om probiotika har någon terapeutisk effekt och kan användas som behandling vid IBS. Metod: Fem randomiserade, dubbelblindade, placebo-kontrollerade studier granskades. De undersökte den terapeutiska effekten av varierande probiotika på IBS-patienter. Artiklarna erhölls genom sökningar i den medicinska databasen PubMed, under februari månad 2016. Resultat: Av dessa fem studier visade fyra på en signifikant symptomförbättring av framförallt buksmärta/obehag. Bäst resultat sågs i en studie som undersökte probiotikan Lactobacillus plantarum 299v. Tre av fem studier visade på en signifikant förbättring av testpersonernas livskvalité. I endast en av studierna, som undersökte effekten av Escherichia coli Nissle 1917, påvisades ingen signifikant skillnad mellan E.coli (probiotika) och placebo. Slutsats: Baserat på de fem studierna föreligger det i nuläget skäliga bevis för att en behandling med specifika probiotika kan ge symtomförbättring av buksmärta/obehag hos IBS-patienter, och förbättrar patienternas livskvalité. Vidare studier krävs för att fastställa den mest effektiva probiotikan, dosen och behandlingslängden. / Background: IBS (Irritable Bowel Syndrome) is a functional gastrointestinal disorder, with an unclear etiology and pathophysiology. IBS is a common disorder in the Western population and is characterized by recurrent abdominal pain/ discomfort, bloating, flatulence, diarrhea and/ or constipation. There is currently no cure for IBS, but the interest in probiotics as an option of treatment has recently increased. Probiotics have been defined as live microorganisms that, when administered in adequate amounts, provide a health benefit on the host, and are believed to have a symptomatic effect in IBS. Probiotics have therefore been of interest for the treatment in IBS. Purpose: The purpose of this study is to examine whether probiotics have a therapeutic effect and if it could be used as a treatment for IBS. Method: Five randomized, double-blind, placebo-controlled studies were reviewed that examined the therapeutic effect of different probiotics in the IBS-patients. Articles were obtained through searches in the medical database PubMed, during the month of February 2016. Results: Four of the five studies showed a significant improvement of symptoms especially in abdominal pain/ discomfort. The best results were seen in a study that investigated the probiotic Lactobacillus plantarum 299v. Three of the five studies showed a significant improvement in quality of life (QOL) of the subjects in the study. One of the five studies which examined the effect of Escherichia coli Nissle 1917, there was no significant difference between E.coli (probiotics) and placebo. Conclusion: There is reasonable evidence that treatment with certain probiotics might provide improvement in symptoms of abdominal pain/ discomfort, and increase patients quality of life based on the five studies. Further studies are required to determine the most effective probiotic, dose and duration of IBS-treatment.

Hospital-acquired infections are to date a major challenge in the patient safety. The proliferation of pathogens such as Staphylococcus aureus, Escherichia coli and Klebsiella pneumoniae is often reported in connection with textiles, which represent a significant source of transmission. This leads most often to the contamination and cross-contamination of the hospitalized patient and the hospital staff. A promising approach and the immediate objective of this research is the application of probiotics to a textile fabric. They provide preliminary evidence in being able to inhibit pathogenic bacteria growth through their competitive mechanism. During this study, screen-printing was used as a method to apply probiotics on a polyester fabric. The viability of probiotics on the fabric was evaluated in the agar plate test method. Samples that exhibited a growth of grown out colonies were further tested regarding their efficacy towards the abovementioned pathogens. This was determined in a competition test, that included the individually inoculation of the samples with the different bacteria strains. Contact-angle measurements and abrasion resistance as well as the durability were tested in order to investigate the applicability of the fabric and scanning electron microscope images were taken to detect probiotics and to evaluate the quality of the print on the fabric. Major findings included that probiotics were able to survive on the polyester fabric and that these viable probiotics are further successful in the growth inhibition of pathogenic bacteria. Thereby their efficacy against pathogens is related to the viability the probiotics. As neither the probiotics used in this study, nor the other ingredients are considered as hazardous, this process is environment-friendly. This work increases the understanding of probiotics mechanism and their survival and competition behaviour on a textile material. It opens a way in the reduction of hospital-acquired infections through the beneficial effects of probiotics.

Submitted in partial fulfillment for the Degree of Master of Applied Sciences in Biotechnology, Durban University of Technology, Durban, South Africa, 2016. / Prebiotics are defined as non-digestible oligosaccharides (NDOs) or polysaccharides (NDPs), which promote the growth of beneficial lactic acid bacteria in the colon. Algae are rich in polysaccharides and can be exploited as prebiotics for functional food ingredients to improve human and animal health. Currently, inulin is the most widely used ingredient in the prebiotics market, which is produced from live plants and requires expensive production processing. There is a vast repository of marine life with algae as a major source of nutrients. Therefore, this study provides an alternative source for prebiotic production and examines marine and freshwater algae that promote the growth of two strains of Lactobacillus delbrueckii subs. (Lactobacillus lactis and Lactobacillus bulgaricus) and one strain of Bifidobacterium spp. (Bifidobacterium longum). Monosaccharides of the oligosaccharide fraction of marine and freshwater algal extracts were investigated with the use of thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) after acidic hydrolysis of cell matrix polysaccharides. A total of fifty-five marine and freshwater aqueous algal extracts were assessed for their effect on the growth of L. lactis, B. longum and L. bulgaricus over a 96 hour period. Relative to the negative control, 34.5% algal extracts showed improved growth on one or more probiotic bacteria. The optimum time for maximum bacterial growth was noted at 48 h for all the tested aqueous algal extracts. Five marine and freshwater algal cultures (Spirulina platensis, Chlorococcum spp., Dunaliella salina, Scenedesmus magnus, Chlorella spp. and algal extract no. 48) from various aquatic environments in Kwa-Zulu Natal showed the best growth dynamics and demonstrated the greatest potential as sources of biomass for prebiotic production. These algal extracts were able to significantly increase the growth of at least one of the three probiotic bacteria (p < 0.05). Aqueous algal extract from S. platensis was regarded as the best algal source for prebiotics as it demonstrated a greater stimulatory effect on the growth of all three probiotic bacteria (L. lactis, B. longum and L. bulgaricus) compared to tested aqueous algal extracts and the inulin used as a positive control. The results obtained by HPLC for characterization confirmed TLC data, as xylose and galactose were detected by both chromatograms. These data indicated that xylose and galactose were present in aqueous algal extracts from S. magnus and S. platensis and galactose in aqueous algal extract no. 48. Xylose was most abundant in aqueous algal extracts from S. platensis (3mg/ml) and S. magnus (2.3mg/ml). In conclusion aqueous algal extracts from S. platensis, Chlorococcum, D. salina, S. magnus, Chlorella and algal extract no. 48 are potential sources for prebiotic production. Spirulina platensis extract was regarded as the best algal source. Xyose and galactose characterized by HPLC in algal extracts make up oligosaccharides that function as prebiotic compounds for stimulation of probiotic bacteria. There is a great scope for successful production of prebiotics from algal sources in South Africa. / M

Ce travail de thèse a permis la mise au point d'un procédé d'encapsulation de bactéries probiotiques dans des matrices ne contenant que des ingrédients laitiers. L'étude à l'échelle laboratoire puis, le dimensionnement du procédé à l'échelle pilote ont permis la production de microparticules stables dans des milieux aqueux et résistantes à des conditions simulant l'estomac. La nature des protéines présentes mais également leurs proportions ont influencé la localisation des bactéries dans les microparticules : à la surface pour une matrice uniquement composée de caséine et à l'intérieur pour une matrice composée à la fois de caséines et de protéines solubles. L'interprétation de ces résultats par une étude à l'échelle moléculaire des interactions qui s'établissent entre les bactéries probiotiques et les protéines laitières a été possible grâce à l'utilisation de la microscopie à force atomique. Les protéines solubles interagissent de façon spécifique avec les bactéries alors que les caséines interagissent de façon non spécifique. De même, la présence de pili à la surface de la bactérie est favorable à la l'établissement d'interactions fortes entre LGG et les protéines solubles. Ce travail a donc permis d'expliquer des résultats obtenus à l'échelle macroscopique (taux d'encapsulation et de survie dans des conditions gastriques) grâce à des observations microscopiques et une étude à l'échelle nanoscopique des interactions entre les différents composants. Cette approche multi-échelle a élucidé certains mécanismes régissant l'encapsulation de bactéries probiotiques dans des matrices laitières / In this work thesis, an encapsulation process for probiotic bacteria using only milk proteins is developed. The laboratory scale followed by a pilot and an industrial scale development of the process allow the production of stable and resistant microparticles both in aqueous and gastric media. The nature and quantities of proteins added is found to influence the bacterial location in the microparticles. The bacteria are located mostly at the surface when only caseins are present. Oppositely, well encapsulated bacteria are observed when addition of whey proteins are performed. A molecular study of interactions established between milk proteins and bacteria is possible with the use of atomic force microscopy. Whey proteins are found to specifically interact with bacteria whereas the caseins establish non-specific links. In addition, the presence of piliated bacteria is found favorable to establish strong and long interactions between proteins and bacteria. This work permits the interpretation of results obtained at a macroscale (encapsulation rate and bacterial survival in stomach conditions) thanks to microscopic observations and nanoscopic interactions study. This multiscale approach permits the elucidation of mechanisms driving the probiotic encapsulation in milk matrices

O presente trabalho teve como objetivo verificar a viabilidade da cepa probiótica Bifidobacterium animalis subsp. lactis Bb-12 incorporado em margarina, suplementada com inulina, concentrado protéico de soro (WPC) e concentrado de caseína (CMP), bem como avaliar as características do produto e a resistência do probiótico às condições simuladas do trato gastrintestinal humano. Foram produzidos 7 diferentes tipos de margarinas de mesa (60% de lipídios: 60 % de óleo de palma + 40% de óleo de canola), empregando-se um modelo de mistura, onde inulina, WPC e CMP foram as variáveis estudadas. Uma formulação controle foi produzida (M8), sem adição desses ingredientes. A utilização da mistura do óleo de palma com óleo de canola favoreceu nutricionalmente as formulações, fornecendo produtos contendo ácidos graxos essenciais em sua composição e ausência de ácidos graxos trans. As formulações M1 a M7, exceto a formulação M2 após o 21º dia de armazenamento, apresentaram populações satisfatórias de Bb-12 para um alimento probiótico, com populações acima de 6 log UFC/g durante 35 dias de armazenamento. Margarinas suplementadas com inulina apresentaram populações satisfatórias durante todo o armazenamento, atingindo populações de 8,01 log UFC/g ao 35º dia (M1). Além disso, M3 e M6, revelaram populações de Bb-12 de 6,87 log UFC/g e 7,27 log UFC/g (dia 35), respectivamente. Por outro lado, M8 não foi caracterizada como margarina probiótica, uma vez que apresentou populações abaixo de 6 log UFC/g, já ao 1º dia de armazenamento. Embora WPC seja utilizado em pesquisas para aumentar a viabilidade de probióticos em alimentos, a suplementação de margarina com WPC sem inulina ou CMP não resultou em populações satisfatórias de Bb-12, apresentando decréscimo de 7,82 (dia 1) para 4,64 log UFC/g (M2, dia 35) (p<0,05). Durante todo o ensaio de resistência in vitro, Bb-12 apresentou sobrevivência significativamente superior (p<0,05) em M1 e revelou populações acima de 6 log UFC/g após 6h de ensaio mesmo ao 28º dia. As populações observadas para M2 diminuíram drasticamente durante o ensaio in vitro (5 log UFC/g após 2h no dia 7). Para as outras formulações, as populações de Bb-12 diminuíram 2 log UFC/g após 2h de ensaio in vitro. Entretanto, M1, M2 e M5 (dias 14 e 28) revelaram aumento significativo nas populações de Bb-12 (p<0,05) entre a fase gástrica (2h) e a segunda fase entérica (6h). As margarinas suplementadas com inulina, principalmente M1, revelaram decréscimo significativo no pH durante todo o armazenamento (p<0,05). Entretanto, isto não afetou a qualidade sensorial dos produtos, uma vez que não foram detectadas diferenças significativas entre as formulações após 7 e 14 dias de armazenamento (p>0,05). A suplementação de margarina com inulina e CMP garantiu populações apropriadas de Bb-12 durante o armazenamento estudado pelo menos até o 28º dia. Além disso, contribuiu para sua sobrevivência durante o ensaio de resistência in vitro. Os resultados revelaram que a margarina apresenta-se como uma matriz alimentar adequada para administração de Bb-12, principalmente quando a inulina foi adicionada. / This study aimed to determine the viability of probiotic Bifidobacterium animalis subsp. lactis Bb-12 incorporated in margarine, with inulin, whey protein concentrate (WPC) and caseinomacropeptide (CMP) supplementation. In addition, the in vitro resistance of Bb-12 incorporated in margarine and related properties were evaluated. Seven margarine-making trials (60% of fat: 60% of palm oil +40% canola oil) were produced, using a mixture model, where inulin, WPC and CMP were the variables studied. Also, a control formulation without these ingredients was manufactured. The use of blending palm oil with canola oil improved the margarine formulations nutritionally, providing products containing essential fatty acids in its composition and absence of trans fatty acids. The formulations M1 to M7, except M2 after 21 days of storage, revealed satisfactory Bb-12 populations for a probiotic food, with counts above 6 log CFU/g during 35 days of storage at 5±1ºC. Margarines supplemented with inulin presented suitable Bb-12 populations throughout the whole storage period, reaching up to 8 log CFU/g by the end of storage (M1). Also, M3 and M6, revealed Bb-12 populations of 6.87 log CFU/g and of 7.27 log CFU/g (day 35), respectively. In contrast, M8 was not characterized as probiotic margarine, since it showed Bb-12 populations below 6 log CFU/g on day 1. Even though whey protein is largely employed in probiotic foods, margarine supplementation with WPC without inulin or CMP did not lead to Bb-12 satisfactory populations, decreasing from 7.82 (day 1) to 4.64 log CFU/g (M2, day 35) (p<0.05). During the whole in vitro assays, Bb-12 survived significantly better (p<0.05) in M1 and revealed populations above 6 log CFU/g after 6h even after 28 days. M2 populations decreased drastically during the in vitro assays for all storage period tested (reduction of 5 log CFU/g after 2h of in vitro assays on day 7 and populations of 2.8 log CFU/g after 6h). For the other formulations, Bb-12 populations decreased 2 log CFU/g after 2h of the in vitro assays. However, for M1, M2 and M5 (on day 14 and 28) the populations of Bb-12 increased significantly (p<0.05) between the gastric phase (2h) and the enteric phase (6h). Formulations containing inulin, mainly M1, showed a significant decrease in pH values during the whole storage period (p<0.05). However, this ingredient did not affect the sensory quality of products, since no significant differences between formulations after 7 and 14 days of storage were observed (p>0.05). The supplementation of margarine with inulin and CMP guaranteed appropriate Bb-12 populations during storage for at least 28 days, and also contributed for its survival throughout the in vitro assays. Therefore, margarine might be considered an appropriate food matrix for Bb-12 survival, mainly when inulin is also added.

A crescente preocupação da população com a saúde tem aumentado a demanda por alimentos que apresentam propriedades funcionais, os quais, além do valor nutricional agregado, estimulam atividades fisiológicas e/ou metabólicas proporcionando benefícios ao indivíduo. Dentre estes encontram-se os alimentos probióticos, prebióticos e simbióticos. Requeijão é um alimento que apresenta potencial como veículo de espécies de micro-organismos probióticos, pois sua matriz apresenta características que contribui para a sobrevivência dos micro-organismo, além de ser um produto com boa aceitabilidade no Brasil. Os alimentos probióticos se destacam no mercado devido às várias evidências científicas sobre seus efeitos benéficos àsaúde humana. Desta forma, o presente trabalho teve como objetivo avaliar a viabilidade celular de 6 cepas de Lactobacillus a saber: L. plantarum ATCC 8014, L. acidophilus ATCC 4356, L. delbrueckii UFV H2B20, L. fermentum ATCC 9338, L. casei ATCC 7469 e L. paracasei SP11 incorporadas no requeijão cremoso. Testes preliminares foram realizados a fim de se adequar a formulação de requeijão cremoso com as características requeridas pela legislação vigente. Posteriormente, fabricou-se o requeijão cremoso probiótico sendo incorporado as respectivas cepas, de forma individual, onde avaliou-se a viabilidade destas cepas ao longo do período de 65 dias de estocagem a 5ºC, tempo este correspondente a vida útil do produto no mercado. Amostras foram coletadas para caracterização físico química (pH e acidez titulável) nos tempos 7, 30, 45 e 65 dias. Posteriormente o requeijão foi caracterizado quanto aos padrões de identidade estabelecido pela ANVISA. No tempo inicial e após 65 dias amostras foram submetidas à análise sensorial sendo avaliados atributos como sabor, odor, aparência e impressão global, bem como a intenção de compra e índice de aceitabilidade. Nas formulações probióticas a população de células viáveis em 1g de requeijão contendo Lactobacillus foi superior a 108 UFC/g após 65 dias de armazenamento a 5ºC estando em conformidade com os padrões estabelecidos pela ANVISA. As formulações contendo Lactobacillus apresentaram valores de pH e acidez titulável mais ácidos em relação a formulação base, atingindo valores de pH inferiores a 6.44, e com produção máxima de ácido láctico de 0,43% no entanto mantiveram-se constantes no período de armazenamento por 65 dias. As análises físico - químicas mostraram que as formulações de requeijão cremoso estão de acordo com o padrão mínimo de identidade do requeijão cremoso estabelecido pela ANVISA. A análise sensorial revelou que a incorporação de Lactobacillus nas formulações não influenciou negativamente nos atributos avaliados, bem como na aceitação do produto, sendo a formulação contendo Lactobacillus paracasei que obteve a maior aceitação, sendo as demais classificadas pelos provadores como \"gostei ligeiramente\". Estes resultados permitem concluir que requeijão cremoso e cepas de Lactobacillus são adequados para a formulação de um novo alimento funcional. Desta forma, contribuiu-se para o desenvolvimento de uma tecnologia para produção de requeijão cremoso probiótico em conformidade com os padrões de qualidade estabelecidos na legislação vigente. / that have functional properties, which, besides the added nutritional value, stimulate physiological and / or metabolic activities providing benefits to the individual. Among these are probiotic, prebiotic and symbiotic foods. Brazilian cream cheese is a food that has potential as a vehicle for probiotic microorganism species, since its matrix has characteristics that contribute to the survival of microorganisms, besides being a product with high acceptability in Brazil. Probiotic foods stand out in the market because of the various scientific evidence on their beneficial effects on human health. Thus, the present work\'s objective was to evaluate the cell viability of 6 strains of Lactobacillus, namely: L. plantarum ATCC 8014, L. acidophilus ATCC 4356, L. delbrueckii UFV H2B20, L. fermentum ATCC 9338, L. casei ATCC 7469 And L. paracasei SP11 incorporated in Brazilian cream cheese. Preliminary tests were performed in order to adapt the Brazilian cream cheese formulation to the characteristics required by current legislation. Subsequently, the probiotic Brazilian cream cheese was inoculated with the respective strains, individually, where the viability of said strains was evaluated over the 65-day storage period at 5 ° C, which corresponds to the shelf life of the product on the market. Samples were collected for physico-chemical characterization (pH and titratable acidity) in 7, 30, 45 and 65 days. Subsequently the Brazilian cream cheese was characterized as to the identity standards established by ANVISA. At the initial time and after 65 days, samples were submitted to sensorial analysis being evaluated attributes such as taste, odor, appearance and overall impression, as well as the purchase intention and acceptability index. In all probiotic formulations the viable cell population in 1g of Brazilian cream cheese containing Lactobacillus was greater than 108 CFU / g after 65 days of storage at 5 ° C, being in accordance with standards stablished by ANVISA. The formulations containing Lactobacillus presented pH values and titratable acidity more acidic in relation to the base formulation, reaching values of pH lower than 6.44 and with maximum production of lactic acid of 0.43%, however, they remained constant in the storage period for 65 days. The physico-chemical analyzes showed that all the Brazilian cream cheese formulations are in accordance with the minimum standard established by ANVISA. Sensory analysis revealed that the incorporation of Lactobacillus in the formulations did not negatively influence the evaluated attributes, as well as the acceptance of the product. The formulation containing Lactobacillus paracasei obtained the most acceptance, being the others classified by the tasters as \"liked slightly\". These results allow us to conclude that Lactobacillus are suitable microorganisms for the creation of a new functional product, as well as the probiotic Brazilian cream cheese. In this way, this work contributed to the development of a technology for the production of probiotic Brazilian cream cheese in compliance with the quality standards established in the current legislation.

Probióticos do gênero Lactobacillus estão sendo amplamente investigados no tratamento da periodontite. Contudo, os efeitos de microrganismos do gênero Bifidobacterium ainda são pouco conhecidos. Este estudo avaliou os efeitos do probiótico Bifidobacterium animalis subsp. lactis (B. lactis) HN019 como adjuvante à raspagem e alisamento radicular (RAR) no tratamento da periodontite experimental (PE) em ratos. No dia 0 do experimento, 32 ratos foram alocados em 4 grupos: C (controle), PROB (probiótico), PE-RAR e PE-RARPROB. Nos grupos PE-RAR e PE-RAR-PROB, a PE foi induzida pela colocação de ligaduras de seda ao redor dos primeiros molares inferiores dos animais. No 14&deg; dia, as ligaduras foram removidas e realizou-se a RAR. Nos animais dos grupos PROB e PE-RARPROB, o probiótico B. lactis HN019 foi administrado diariamente (10 mL/dia de 109 unidades formadoras de colônia) por 15 dias tendo seu início no 14&deg; dia do experimento. Os animais de todos os grupos foram submetidos à eutanásia 29 dias após o início do experimento. As hemimandíbulas e amostras de intestino delgado foram coletadas. Foram realizadas análises histomorfométricas, microtomográficas e imunohistoquímicas. Foram investigados, também, os efeitos microbiológicos de B. lactis HN019 no biofilme associado às ligaduras durante o desenvolvimento da PE. Todos os dados foram analisados estatisticamente. O Grupo PE-RAR-PROB apresentou menores reabsorção óssea alveolar e perda de inserção conjuntiva quando comparado ao Grupo PE-RAR, bem como menor número de osteoclastos, maior expressão de citocinas anti-inflamatórias e menor expressão de citocinas pró-inflamatórias (p <0,05). No grupo PE-RAR-PROB, os valores médios da profundidade da cripta do jejuno e duodeno foram significativamente maiores que aqueles do grupo PE-RAR. A proporção de bactérias aeróbias/anaeróbias foi maior nas amostras de biofilme de animais tratados com B. lactis HN019 em relação àquelas de animais não tratados (p <0,05). Dentro dos limites deste estudo, pode-se concluir que a utilização de B. lactis HN019 como adjuvante à RAR promove benefícios histológicos, microtomográficos e imunológicos adicionais no tratamento da PE em ratos, bem como melhora a morfologia intestinal. / Lactobacillus probiotics have been investigated in periodontitis. However, the effects of the genus Bifidobacterium on periodontitis are hardly known. This study evaluated the effects of the probiotic Bifidobacterium animalis subsp. lactis (B. lactis) HN019 as an adjunct to scaling and root planing (SRP) in rats with experimental periodontitis (EP). At baseline, 32 rats were assigned to 4 groups: C (control), PROB, EP-SRP and EPSRP- PROB. In groups EP-SRP and EP-SRP-PROB, the mandibular first molars of the animals received a ligature. At day 14, the ligatures were removed and SRP was performed. Animals of groups PROB and EP-SRP-PROB were orally administered with 10 mL/day of 109 colony forming units of B. lactis HN019 for 15 days, starting at day 14. Animals were euthanized at day 29. The jaws and samples of the duodenum, jejunum, and ileum were resected. Histomorphometric, microtomographic and immunohistochemical analyses were performed. Microbiological effects of B. lactis on biofilm were also evaluated. Data were statistically analyzed. Group EP-SRP-PROB presented reduced alveolar bone resorption and attachment loss when compared with Group EP-SRP (p<0.05). Group EP-SRP-PROB showed significantly fewer osteoclasts, increased expression of anti-inflammatory cytokines and reduced expression of proinflammatory cytokines compared with Group EP-SRP (p<0.05). In group EP-SRPPROB, the mean values of crypt depth of the jejunum and dudoenum were significantly higher than the ones from group EP-SRP. B. lactis promoted a higher ratio between aerobic and anaerobic bacteria in biofilm samples (p<0.05). Within the limits of this study it can be concluded that the use of B. lactis HN019 as an adjunct to SRP promotes additional histologic, microtomographic and immunologic benefits in the treatment of EP in rats and improves the intestinal morphology.

O objetivo deste trabalho foi avaliar in vitro os mecanismos de ação coagregação, produção de peróxido de hidrogênio e ácidos orgânicos por probióticos no combate ao crescimento de Candida spp. A Estomatite sob Prótese é uma das infecções que acomete a mucosa oral, nas áreas recobertas por próteses, como resultado de condições locais ou sistêmicas que alteram a microbiota do hospedeiro, podendo estar associada à presença de Candida. O tratamento com antifúngicos tópicos é amplamente difundido para tratar candidose, porém algumas cepas de Candida se tornaram resistentes aos fármacos mais utilizados devido ao seu uso indiscriminado. Alguns trabalhos mostraram inibição do crescimento de Candida por probióticos, mas não se sabe ao certo quais mecanismos de ação agem nesta inibição. Para esta investigação foram utilizadas 77 amostras de leveduras isoladas de usuários de próteses totais, 4 cepas Candida ATCC, e 3 cepas de Lactobacillus. Realizou-se os testes de inibição de Candida por probióticos, produção de peróxido de hidrogênio e ácidos orgânicos, coagregação, curva de crescimento e inibição por ácido láctico. Os resultados demostraram a produção de H2O2 pelos lactobacilos, porém este mecanismo não pôde ser destacado como o responsável pela inibição. Os índices de coagregação obtiveram coeficiente de correlação r =0,5 com a capacidade de inibição dos micro-organismos. Houve intensa produção de ácidos orgânicos em culturas e co-culturas, de acordo com a cepa de lactobacilo presente. A presença de Candida não alterou as contagens de UFC dos lactobacilos. Já a presença de lactobacilos provocou redução nas contagens de Candida, especialmente C. albicans e C. dubliniensis. As curvas de crescimento e de pH mostraram que o comportamento das co-culturas é semelhante ao das culturas de lactobacilos. A análise de pH mostrou que a acidificação ocorreu em todas as culturas, porém L. rhamnosus promoveram uma maior acidificação. Concluímos que o ácido láctico é o mecanismo de ação mais importante dentre os estudados, sendo capaz de reduzir a contagem de UFC e alterar as curvas de crescimento de C. albicans e C. dubliniensis e que a inibição ocorre através de múltiplos fatores que podem variar de acordo com as espécies de leveduras e lactobacilos presentes. / This work objective was to evaluate in vitro the action mechanisms of coaggregation, hydrogen peroxide production and organic acids in preventing the growth of Candida spp.. The denture stomatitis is one of the infection types that affect the oral mucosa, in the area covered by prosthesis as a result of local or systemic conditions that changes the bacterial microbiota of the hostess, usually associated with the presence of Candida. The treatment using topical antifungal agents is widely spread to treat candidosis. However, some of the Candida strains became resistant to the most used drugs because of its indiscriminate use. Some pieces of work revealed success in preventing the growth of Candida by probiotics, but it is still unknown which mechanisms act on this exhibition. For this investigation there have been used 77 yeast samples isolated from total prosthesis users, being 4 ATCC strains of Candida, and 3 strains of Lactobacillus. There have been run inhibition tests of Candida by probiotics, hydrogen peroxide and organic acids production, coaggregation, growth curve and lactic acid inhibition. The results detect the H2O2 production by lactobacilli; however this mechanism could not be highlighted as the responsible for the inhibition observed. The coaggregation indexes showed correlation coefficients of r = 0.5 with the microorganisms inhibition capacity. There has been intensive organic acid production in cultures and co-cultures, according to the present lactobacilli strain. Candida presence did not alter the lactobacilli UFC count, whereas the lactobacilli presence caused reduction in Candida count, especially C. albicans and C. dubliniensis. The growth curves and pH showed that the behavior of co-cultures is similar to the lactobacilli cultures. The pH analysis showed that acidification occurred for all cultures; nevertheless, L. rhamnosus has promoted a greater acidification. We concluded that lactic acid is the most important mechanism among the studied ones, being capable of reducing UFC count and shifting the growth curves for C. albicans and C. dubliniensis. Furthermore, we concluded that the inhibition occurs through multiple factors that may vary according to the present yeast and lactobacilli species.

O trabalho teve como objetivos desenvolver sorvetes com leite de cabra, polpa de cajá e cepas potencialmente probióticas Lactobacillus rhamnosus HN001 e Lactobacillus paracasei LBC82, verificar a viabilidade dessas cepas e avaliar as características dos produtos ao longo de seu armazenamento a -18ºC por até 12 semanas, bem como a resistência das cepas incorporadas nos produtos a condições gástricas e entéricas simuladas in vitro. Primeiramente, foi desenvolvido creme de leite de cabra para aplicação no produto. Em seguida, foram realizados os ensaios de desenvolvimento dos sorvetes, os quais variaram de acordo com a adição de gordura láctea caprina (sorvetes SC) e a sua substituição por inulina (sorvetes SI) e com a adição de 2 cepas de Lactobacillus - L. rhamnosus HN001 (Lr) e L. paracasei LBC82 (Lp), tendo sido assim denominados: SCr - com gordura láctea caprina + Lr; SCp - com gordura láctea caprina + Lp; SIr - com inulina + Lr; SIp - com inulina + Lp. Os sorvetes foram armazenados a -18±3ºC por até 12 semanas (84 dias) e analisados quanto às seguintes características: overrun (durante a elaboração), fração de derretimento e dureza instrumental (após 14 dias), composição centesimal (após 21 dias), pH (após 1, 28, 56 e 84 dias), viabilidade dos probióticos (durante o processamento e após 1, 7, 14, 21, 28, 42, 56 e 84 dias) e sua resistência às condições gástrica e entérica simuladas in vitro (após 28 e 84 dias). Os sorvetes foram, ainda, comparados quanto à sua aceitação sensorial (escala hedônica de 9 pontos, com 1 = desgostei muitíssimo e 9 = gostei muitíssimo) na Embrapa Caprinos e Ovinos, Sobral, CE (após 7, 42 e 84 dias) e, para efeito de comparação entre consumidores de dois locais e público distintos, também foram submetidos à análise sensorial, 7 dias após a sua produção, na FCF/USP, SP. Todos os sorvetes, apesar dos valores de pH inferiores a 4,5, apresentaram populações médias de Lactobacllus spp. acima de 8 log UFC/g e estáveis, durante 84 dias de armazenamento. Por outro lado, as cepas testadas mostraram baixa sobrevivência nos testes in vitro realizados, com ligeiro aumento dessa sobrevivência na presença de inulina aos 28 dias de armazenamento, particularmente para SIp. Os 4 sorvetes estudados apresentaram notas médias que variaram de 6,9 (SIr, aos 42 dias) a 8,0 (SCp, aos 7 dias) na avaliação sensorial, sem diferenças significativas entre os sorvetes, mas com uma diminuição significativa nas notas ao longo do armazenamento para os sorvetes com creme (p<0,05), o que não foi observado para os sorvetes com inulina, que mostraram, ainda, maior firmeza (p<0,05), com teores inferiores de lipídeos. Quando os 2 públicos de regiões distintas foram comparados quanto à aceitação sensorial, não foi verificada diferença significativa para os sorvetes com inulina, enquanto que aqueles com creme de cabra obtiveram notas significativamente inferiores (p<0,05) do público de SP (médias de 7,60 e 7,46, respectivamente, para SCr e SCp), comparado àquele do CE (médias de 7,95 e 8,03, respectivamente, para SCr e SCp). Conclui-se que os sorvetes de leite de cabra com polpa de cajá desenvolvidos apresentaram uma boa sobrevivência das cepas de Lactobacillus estudadas no produto e boas características sensoriais. As formulações contendo inulina seriam a melhor opção, por apresentarem, ainda, características prebióticas, teores inferiores de lipídeos, com maior estabilidade quanto à aceitação sensorial com o armazenamento e quando avaliados pelo público dos 2 diferentes locais testados. Os sorvetes estudados são uma opção viável para empregar alimentos como o leite de cabra e a polpa de cajá no desenvolvimento de produtos que contenham características funcionais, portanto, alto valor agregado. / This study aimed to develop a goat milk-based ice-cream with yellow mombin pulp and supplemented with the potentially probiotic strains Lactobacillus rhamnosus HN001 and Lactobacillus paracasei LBC82, check the viability of these strains and evaluate the products\' features throughout their storage at -18ºC for up to 12 weeks, besides the resistance of the strains incorporated into the products through in vitro gastric and enteric simulated conditions. Firstly, a goat milk cream was developed to be used in the product. Next, tests were conducted for the ice-cream formulations developed. These formulations varied according to the addition of goat milk cream (ICMC) or substitution of milk cream by inulin (ICI) and with the addition of two strains of Lactobacillus - L. rhamnosus HN001 (Lr) and L. paracasei LBC82 (Lp), and were designated as follows: ICMCr - with goat milk cream + Lr; ICMCp - with goat milk cream + Lp; ICIr - with inulin + Lr; ICIp - with inulin + Lp. The ice-creams were stored at -18±3°C for up to 12 weeks (84 days) and analyzed for the following features: overrun (during production), melting rate and instrumental hardness (after 14 days), chemical composition (after 21 days), pH (after 1, 28, 56, and 84 days), probiotics viability (during production and after 1, 7, 14, 21, 28, 42, 56, and 84 days) and their resistance to in vitro simulated gastric and enteric conditions (after 28 and 84 days). The ice-creams were also compared for their sensory acceptability (9-point hedonic scale, with 1 = dislike extremely and 9 = like extremely) at Embrapa Sheep and Goats, Sobral, CE (after 7, 42, and 84 days). In order to compare sensory acceptability between consumers and local public from two different areas, sensory analysis was also conducted at the Faculty of Pharmaceutical Sciences at USP, SP, seven days after their production. The four kinds of ice-creams, even though with pH values below 4.5, showed Lactobacllus spp. mean populations above 8 log CFU/g and stable up to 84 days of storage. Nevertheless, the strains tested showed low survival through the in vitro tests and a slight increased survival in the presence of inulin at 28 days of storage, particularly for the ICI ice-creams. The four products studied had mean scores ranging from 6.9 (ICIr, at 42 days) up to 8.0 (ICMCp, at 7 days) in the sensory evaluation, with no significant differences between the ice-creams, but with a significant decrease in the scores during storage for the ice-creams with goat milk cream (p <0.05). This was not observed for the ice-creams with inulin, which also showed higher firmness (p <0.05), with lower lipids levels. When the consumers of the two different areas were compared regarding sensory acceptability of ice-creams, no significant difference was found among ice-creams with inulin, whereas those with goat milk cream received significantly lower scores (p <0.05) from SP consumers (mean 7.60 and 7.46, respectively, for ICMCr and for ICMCp), compared to those of CE (mean 7.95 and 8.03, respectively, for ICMCr and ICMCp). In conclusion, the goat milk-based ice-cream with yellow mombin pulp developed showed good survival of the Lactobacillus strains studied in the product and good sensory features. Formulations containing inulin would be the best option, as they also presented prebiotic characteristics, lower lipid levels, showed greater stability regarding sensory acceptability during storage and when evaluated by the public from the two different areas tested. The ice-creams studied are good options for employing foods like goat milk and yellow mombin pulp in the development of products with functional properties and, therefore, with high added values.