Dissertations / Theses on the topic 'Probiotics'

Il primo obiettivo di questa tesi di dottorato è stata la formulazione di una mozzarella Fiordilatte simbiotica attraverso un rivestimento di alginato di sodio commestibile come veicolo di probiotici (Lactobacillus rhamnosus ) e di frutto-oligosaccaridi prebiotici (FOS). Gli obiettivi di questo studio sono stati la valutazione della capacità della tecnica di rivestimento di intrappolare il microrganismo senza alterarne la vitalità e il monitoraggio della vitalità microrganismo durante l'intero periodo di conservazione . A questo scopo , prove di shelf life sono state condotte a tre diverse temperature di stoccaggio (a 4°C, la normale temperatura di refrigerazione, a 9°C in condizioni di moderato abuso termico e a 14°C, in condizioni di abuso termico) per valutare la qualità microbiologica, sensoriale e funzionale della mozzarella. Pertanto, sono stati monitorati sia i microorganismi di deterioramento e sia i batteri probiotici. I risultati hanno dimostrato che la vitalità dei batteri lattici presenti nel prodotto funzionale è rimasta oltre il limite imposto (107 CFU/g) per l'intero periodo di conservazione in tutte le prove sperimentali con i valori intorno a 4,52 x 107 CFU/g a 4°C, 3,42 x 107 a 9°C e 4,62 x 107 a 14°C. Inoltre , l'aggiunta di sostanze probiotiche e prebiotiche nel rivestimento ha provocato un leggero effetto antimicrobico contro Pseudomonas spp. ed Enterobacteriaceae, che ha a sua volta migliorato il sapore finale del prodotto e prolungato la sua durata commerciale. Il secondo obiettivo di questa tesi è stata l'ottimizzazione di hamburger di manzo arricchiti con frutto-oligosaccaridi (FOS) , inulina e schiuma proteica a base di crusca d’avena. Le diverse formulazioni di hamburger di manzo funzionali sono state ottimizzate dal punto di vista sensoriale mediante un panel addestrato composto da dieci membri . Una volta ottimizzata la formulazione da un punto di vista sensoriale, è stata valutata l'influenza degli ingredienti prebiotici sulle caratteristiche chimiche, fisiche, nutrizionali e tecnologiche degli hamburger di carne. Dai risultati, si è constatato che la combinazione sia dei FOS che dell’inulina, con la schiuma a base di crusca può risultare una strategia interessante per minimizzare la perdita di composti prebiotici durante la cottura. La concentrazione di fibre in tutti i campioni di carne sviluppati, è risultata essere molto superiore al livello minimo imposto (3 g di fibre per 100 g di prodotto alimentare) per un alimento prebiotico. I risultati hanno dimostrato anche che l’aggiunta delle sostanza prebiotiche ha migliorato le caratteristiche tecnologiche e sensoriali degli hamburger. ABSTRACT IN ENGLISH The first objective of this PhD thesis was the formulation of a symbiotic Fiordilatte cheese with an edible sodium alginate coating as carrier of probiotic (Lactobacillus rhamnosus) and prebiotic fructo-oligosaccharides substances (FOS). The objectives of this study are the evaluation of the coating technique to entrap the microorganism without altering its vitality and the monitoring of the microorganism vitality during the entire storage period. To this aims, shelf life tests were conducted at three different storage temperatures (4°C as refrigeration temperature, 9°C as moderate thermal abuse and 14°C as thermal abuse) to assess the microbiological, sensory and functional quality of cheese. Therefore, both spoilage and probiotic bacteria were monitored. Results proved that viability of lactic acid bacteria in the functional product remained over the imposed limit (107 CFU/g) for the entire storage period in all the experimental trials with counts accounting for about to 4.52 x 107 CFU/g at 4°C, 3.42 x 107 at 9°C and 4.62 x 107 at 14°C. Furthermore, the addition of probiotic and prebiotics substances in the coating provoked a slight antimicrobial effect against Pseudomonas spp. And Enterobacteriaceae that improved the final taste of the product and prolonged its shelf life. The second goal of this thesis the optimization of beef burgers enriched with fructo-oligosaccharides (FOS), inulin and an oat bran loaded protein foam. The various formulations of functional beef burgers were optimized from a sensory point of view by means of a panel of ten trained members. Once the formulation has been optimized, the influence of the prebiotic ingredients on chemical, physical, nutritional and technological characteristics of meat burgers was assessed. From the results, it was found that combinations of both FOS and inulin, respectively combined with oat bran loaded foam are an interesting way to minimize the loss of prebiotic compounds during cooking. The concentration of fibers was found to be much higher than the minimal imposed level (3 g of dietary fiber for 100 g of food product) for prebiotic food, in all developed meat samples. Results also suggest that prebiotic addition to meat also improved the technological and sensory characteristics of investigated burger.

Syfte: Syftet med studien var att inventera lactobacillanvändandet i vården av patienter på Akademiska sjukhuset i Uppsala. Metod: Enkäter skickades ut till 67 vårdavdelningar på sjukhuset. Svarsfrekvens var 79 % och bortfallet 21 %.

Resultat: Huvudresultatet i studien är att en stor del (76 %) av avdelningarna vid Akademiska sjukhuset i Uppsala använder sig av lactobaciller i vården av patienter. Den mest frekvent använda produkten är Proviva saft/fruktsoppa (54 %). De samlade filprodukterna står tillsammans för 24 % av användandet. Den främsta indikationen varför lactobacillprodukter ges är vid antibiotikabehandling för att undvika relaterade magproblem. Nästa stora indikationsgrundande tillstånd är den generella användningen då man serverar exempelvis filprodukter på generell basis ("det enda som finns”).

Diskussion: Att användningen av Lactobaciller är så utbredd kan bland annat vara ett resultat av intensiv marknadsföring som tilltalat både vårdpersonal och patienter. Vid några av de adminsitreringsgrundande tillstånden som avdelningarna angett finns kliniska studier som kan tänkas ligga till grund för administration, men mycket av distributionen tycks ske utan evidens som grund.

Slutsats: Lactobaciller används i stor utsträckning i vården av patienter på Akademiska sjukhuset och vid många olika tillstånd, men med tonvikt på olika mag-tarm åkommor och främst vid antibiotikabehandling.

The presented diploma thesis is focused on the preparation and the monitoring of the biological stability of beer enriched with probiotics. Probiotic bacterial strains of Lactobacillus acidophilus, Bifidobacterium breve and Bifidobacterium bifidum were used in this study. The theoretical part is divided into two sections. In the first section, probiotics are generally characterized, and their role within a gut microbiota is described. Next, the microbiology of particular probiotic microorganisms including the genera of Lactobacillus and Bifidobacterium is described. Then, some factors influencing the viability and the growth of probiotics are stated. In this section, biological effects of probiotics on the human organism and their potential clinical applications are described. The second section of the theoretical part deals with the technology of brewing, the chemical composition of beer and particular beer styles. In the experimental part, the methods of probiotic bacterial cell concentration and viability determination were optimized. Several techniques for determination of these parameters were selected, particularly the cultivation method, flow cytometry and the spectrophotometric measurement of turbidity. Then, the growth curves of the probiotic strains were measured in MRS medium. Probiotic bacteria were cultivated in model beer samples, i.e. in MRS media with several different concentrations of ethanol. It is possible to say that ethanol did not have significant effect on probiotics growth. Next, experimental cultivations of individual probiotic bacteria and their mixtures in nine real beer samples were conducted. No increase of viable cells concentrations was detected in the samples. On the contrary, a decrease of the concentrations was observed, mainly in the samples with individual bacterial strains. However, certain values of viable cells concentrations were determined at the end of the cultivations in all cases. A pale, top-fermented beer was brewed and supplemented with probiotics, and the concentrations of viable probiotic cells were monitored during 37 days of fermentation. A decrease of concentrations by two orders of magnitude of CFU/ml was observed in almost all samples. Yet, viable cells of probiotic bacteria were detected in all samples of beer at the end of the fermentation. Maximal concentration of viable probiotic cells in the brewed beer was determined with the cultivation method at (3,80 ± 0,14)10^5 CFU/ml. Chosen samples were analyzed with HPLC-RI method that quantified the common beer concentrations of ethanol in all chosen samples, lactic acid was not detected. Sensory analysis was conducted as well. Based on the results of the experimental part and the bibliography, an optimal technology of the preparation of beer enriched with probiotics is discussed in this study.

Thesis (MSc Nutrition Science)--Stellenbosch University, 2004.
ENGLISH ABSTRACT: Probiotics (live microbes) and prebiotics (non-digestible food-ingredients) are rapidly gaining interest worldwide as supplements and functional food ingredients but little South African information in this regard is available. Furthermore, the availability of South African produced probiotic concentrates for commercial use is also very limited. The aims of this study therefore were to complete a market and product assessment of probiotic and prebiotic containing products in South Africa and to evaluate probiotic strains for commercial use in South Africa. For the purposes of market and product assessment probiotic and/or prebiotic containing products manufactured in South Africa were identified. The scientific and legal correctness of health and content claims made on the labels of the products were assessed. An exploratory survey was conducted to determine the awareness of South African consumers of probiotics and prebiotics. For the evaluation of probiotic strains for potential commercial use in South Africa, a panel of twelve lactic acid bacteria (LAB) were screened for inhibitory activity against two porcine pathogens and indicator strains from the LMG-panel isolated from the faeces of patients diagnosed with AIDS. The five LAB with the best inhibitory activity were tested for growth in soymilk-base and for the effect of lyophilization on the inhibitory activity thereof. The effect of prebiotics on the growth and inhibitory activity of the strains was tested in vitro. A range of products containing probiotics and prebiotics available on the South African market was identified. Irregulatories concerning health claims on the labels were found, but content claims seemed to be less of a problem. The results also indicate that the proposed South African regulations for the labelling of probiotic and prebiotic containing products need to be revised to include the probiotic and prebiotic related health claims for which sufficient scientific evidence is available. The probiotic strains with potential for commercial use in South Africa that were identified, include Lactobacillus plantarum 423, Lactobacillus casei LHS, Lactobacillus salivarius 241, Lactobacillus curvatus DF38 and Pediococcus pentosaceus 34. These strains were grown successfully in soymilkbase and lyophilization did not have a negative effective on the inhibitory activity thereof. The growth and inhibitory activity of the five LAB were promoted when combined with 1% (w/v) Raftilose® Synergyl . It is concluded that although a variety of probiotic and prebiotic containing products are available on the South African market, the scientific and legislative correctness of especially health related claims is not satisfactory and that South African consumer awareness of these products is low. It is also concluded that a combination of at least three of the five identified LAB and 1% Raftilose® Synergy can be used by South African manufacturers for the production of probiotic and prebiotic containing supplements.
AFRIKAANSE OPSOMMING: Die belangstelling in probiotika (lewendige mikrobe) en prebiotika (onverteerbare voedselbestanddele) as supplemente en funksionele voedselbestanddele is besig om wêreldwyd toe te neem, alhoewel weinig Suid-Arikaanse inligting in hierdie verband beskikbaar is. Die beskikbaarheid van Suid-Afrikaans geproduseerde probiotika konsentrate vir kommersïele gebruik is ook baie beperk. Die doelwitte van hierdie studie was dus om 'n mark- en produkevaluering van probiotika- en prebiotika-bevattende produkte in Suid-Afrika uit te voer en om probiotiese stamme te evalueer vir uiteindelike kommersiële gebruik in Suid-Afrika. Vir die doel van die mark- en produkevaluering is probiotika- en prebiotika-bevattende produkte wat in Suid-Afrika vervaardig word geïdentifiseer. Die wetenskaplike en wetlike korrektheid van die gesondheids- en inhoudsaansprake op die etikette van die produkte is evalueer. 'n Markopname is uitgevoer om die bewustheid van Suid-Afrikaanse verbruikers van probiotika en prebiotika vas te stel. Vir die evaluering van probiotiese stamme vir potensïele kommersiële gebruik in Suid-Afrika is 'n paneel van twaalf melksuurbakteriëe getoets vir inhibitoriese aktiwiteit teen twee patogene geïsoleer uit varke asook teen indikator stamme van die LMG-paneel. Die vyf melksuurbakteriëe met die beste inhibitoriese aktiwiteit is getoets vir groei in sojamelk-basis en ook vir die effek van vriesdroging op die groei en inhibitoriese aktiwiteit van die stamme daarvan. Die effek van prebiotika op die groei en inhibitoriese aktiwiteit van die stamme is in vitro getoets. 'n Reeks van probiotika- en prebiotika-bevattende produkte wat beskikbaar is op die Suid-Afrikaanse mark, is geidentifiseer. Ongeruimdhede met die gesondheidsaansprake op die etikette is gevind, maar inhoudsaansprake was minder problematies. Die resultate dui ook daarop dat die voorgestelde Suid- Afrikaanse regulasies vir die etikettering van probiotika- en prebiotika-bevattende produkte hersien moet word om al die probiotika- en prebiotika-verwante gesondheidsaansprake waarvoor voldoende wetenskaplike bewyse beskikbaar is in te sluit. Die probiotiese stamme met potensiaal vir kommersiële gebruik in Suid-Afrika sluit die volgende in: Lactobacillus plantarum 423, Lactobacillus casei LHS, Lactobacillus salivarius 241, Lactobacillus curvatus DF 38 en Pediococcus pentosaceus 34. Hierdie stamme is suksesvol gekweek in sojamelk-basis en vriesdroging het nie' n negatiewe effek op die groei en inhibitoriese aktiwiteit daarvan gehad me. Die kombinasie van die vyf melksuurbakteriëe met 1% Raftilose® Synergy het die groei en inhibitoriese aktiwiteit daarvan bevorder. Die gevolgtrekking wat gemaak word is dat alhoewel 'n varrasie van probiotika- en prebiotikabevattende produkte beskikbaar is op die Suid-Afrikaanse mark, die wetenskaplike en wetlike korrektheid van spesifiek die gesondheids-verwante aansprake op die etikette daarvan nie bevredigend is nie en dat die bewustheid van die Suid-Afrikaanse verbruikers van hierdie produkte laag is. Die gevolgtrekking kan ook gemaak word dat 'n kombinasie van ten minste drie van die vyf geïdentifiseerde melksuurbakteriëe en 1% Raftilose® Synergy deur Suid-Afrikaanse vervaardigers gebruik kan word vir die vervaardiging van produkte wat probiotika en prebiotika bevat.

Počet obyvatel trpících různými infekčními, zánětlivými a alergickými nemocemi stejně jako výskyt laktózové nesnášenlivosti a vysoké hodnoty krevního cholesterolu, má narůstající tendenci. Některé z těchto zdravotních problémů jsou způsobeny nevyváženou střevní mikroflorou. Probiotika jsou pak chápána (nejen) jako potravní komponenty, které přispívají k ustanovení mikrobiální rovnováhy (Parker, 1974) mezi zdraví prospěšnými a škodlivými bakteriemi. Z tohoto důvodu, terapie založená na podávání probiotik pacientům přitáhla zájem ze strany vědců. Vhodný probiotický kmen se pak volí v závislosti na požadovaném zdravotním účinku (příp. zdravotním problému, který má být probiotickou terapií léčen). Lactobacillus plantarum 299v již prokázal své blahodárné účinky na lidech a zároveň byla i potvrzena jeho zdravotní bezpečnost, díky čemuž může tato bakterie být kategorizována jako probiotický kmen (Probi AB, Sweden). I díky tomu je Lactobacillus plantarum 299v ve značné oblibě přidáván do mnoha fukčních potravin a prodáván na trhu pod různými jmény, probiotický nápoj ProViva je jedním takovým příkladem. Cílem této práce bylo studovat fermentační proces na žitném, ječmenném a sojovém substrátu pomocí kmene Lactobacillus plantarum 299v, přičemž zvýšená pozornost byla věnována právě soji a ječmeni jako potenciálně novým substrátům pro výše uvedenou bakterii. Hlavními záměry bylo zkoumání růstu a metabolické aktivity bakterie Lactobacillus plantarum 299v v asociaci s různými cereálními substráty, a později bylo studováno totéž také ve směsi fermentované cereální komponenty s běžně dostupným ovocným džusem. K tomu, aby se dosáhlo optimálních podmínek fermentace, je třeba vzít v úvahu několik aspektů. Hlavní role při konceptování nového fermentovaného produktu patří především zpracování a taktéž kompozici surového materiálu, růstové kapacitě a produktivitě bakteriální kultury a stabilitě finálního produktu během skladování (De Vuyst, 2000). Tyto parametry jsou důležité hlavně ze strany výrobců. Krom toho jsou tu ale i zákazníci, pro něž je přijatelnost produktu založena z velké části na organoleptických vlastnostech finálního probiotického produktu, tj. aromatu a chuti. Přítomnost a dostupnost různých jednotlivých nutrientů, která byla obsažena ve fermentačním médiu výsledkem rozdílných použitých cereálních substrátů, pravděpodobně vyústila v odlišnosti metabolických drah, což pak později mohlo způsobit rozdíly v organoleptických vlastnostech finálního produktu.

The development of childhood atopy is postulated to be influenced by perinatal microbial exposure. Inadequate numbers of probiotic organisms forming the intestinal microbiota, is an early pre-symptomatic feature linked to the expression of allergy. Perinatal probiotic supplementation is therefore hypothesized to contribute to the microbiota mediated immunomodulatory response determining immune hypo- responsiveness to allergens. Aims: This thesis examined the anti-allergy immunomodulatory effects of probiotic bacteria invitro in the context of the pathophysiology of atopy, as a preamble to perinatal probiotic supplementation. Methods: Cytokine responses were measured after umbilical cord blood mononuclear cells (CBMCs) were co-cultured in the presence of a consortium of probiotic organisms consisting of two strains of lactobacilli and two strains of bifidobacteria. Additionally pregnant women with a familial history of allergic disease from 36 weeks gestation and their infants to age 6 months, were supplemented with probiotics or a placebo daily. The immunomodulatory effects of probiotics on immune function in vivo was then analysed within the peripheral blood of the supplemented neonates. Results: CBMCs co-cultured in the presence of the probiotic consortium generated a dose dependent, monocyte mediated release of the pro-inflammatory cytokines TNF-a, IL- 12p70, IFN-y and the immunosuppressive cytokines IL-10 and TGF-pi. The consortium down regulated PH A and SEB induced IL-13 (a key allergy orchestrating cytokine) while potentiating IFN-y (a key Thl driving cytokine) responses from CBMCs. Interestingly in the probiotic supplemented group the cord blood eosinophil count was significantly reduced. Additionally the IL-12p70 concentrations in microbial stimulated venous blood at age 6 months was significantly elevated in comparison to the placebo supplemented group. Conclusion: The immunomodulatory effect of probiotic bacteria is marked by a capacity to promote a Thl orientation of the immune system. Probiotics administered during pregnancy and early infancy may therefore be effective in the prevention of Th2 mediated atopic disorder.

Starter cultures of different bacteria strains like lactic acid producing bacteria, Staphylococcus and Kocuria are used when making salami. Starter cultures give the sausage specific flavours and improve the quality and ripening of the final product. Probiotic strains can also be added during the production of salami. Studies have shown that probiotics are good for health and are therefore added to food, such as fermented sausages. In order to work as a probiotic strain, the bacteria have to survive during the production process, storage and through the whole human gastrointestinal tract. The aim of this study was to trace the probiotic strains Lactobacillus casei and Lactobacillus paracasei in salami samples to see if they had survived the production process. Methods used were DNA extraction, PCR, colony PCR and gel electrophoresis. Out of 100 samples in duplicate run in PCR, probiotics were found in only 3 of them. To see if screening of probiotics directly from plates was possible, a colony PCR was done. Colony PCR was made on colonies of two different strains of Lactobacillus casei, Lactobacillus paracasei and Lactobacillus sakei. From each bacteria strain, 5 colonies were analysed. Result showed that colony PCR, to screen for probiotic is a possible method.

Introduction: Ageing has often been associated with a marked reduction in numbers of bifidobacteria and an increase in numbers of clostridia. In addition, ageing is associated with alterations in immunity and poor response to vaccination. Pre- and probiotics are suggested to restore the gut micro flora ecosystem and modulate immune function. The current study evaluated the influence of a novel probiotic, Bifidobacterium longum by. infantis CCUG 52486, with a potential prebiotic (glucooligosaccharide; GIOS) on the gut microbiota composition and on innate immunity in young (18-35y) and older (60-85y) subjects undergoing influenza vaccination.

The role of the human gut microbiota in impacting host’s health has been widely studied in the last decade. Notably, it has been recently demonstrated that diet and nutritional status are among the most important modifiable determinants of human health, through a plethora of presumptive mechanisms among which microbiota-mediated processes are thought to have a relevant role. At present, probiotics and prebiotics represent a useful dietary approach for influencing the composition and activity of the human gut microbial community. The present study is composed of two main sections, aimed at elucidating the probiotic potential of the yeast strain K. marxianus B0399, as well as the promising putative prebiotic activity ascribable to four different flours, naturally enriched in dietary fibres content. Here, by in vitro studies we demonstrated that K. marxianus B0399 possesses a number of beneficial and strain-specific properties desirable for a microorganism considered for application as a probiotics. Successively, we investigated the impact of a novel probiotic yoghurt containing B. animalis subsp. lactis Bb12 and K. marxianus B0399 on the gut microbiota of a cohort of subjects suffering from IBS and enrolled in a in vivo clinical study. We demonstrated that beneficial effects described for the probiotic yoghurt were not associated to significant modifications of the human intestinal microbiota. Additionally, using a colonic model system we investigated the impact of different flours (wholegrain rye and wheat, chickpeas and lentils 50:50, and barley milled grains) on the intestinal microbiota composition and metabolomic output, combining molecular and cellular analysis with a NMR metabolomics approach. We demonstrated that each tested flour showed peculiar and positive modulations of the intestinal microbiota composition and its small molecule metabolome, thus supporting the utilisation of these ingredients in the development of a variety of potentially prebiotic food products aimed at improving human health.

The role of the human gut microbiota in impacting host’s health has been widely studied in the last decade. Notably, it has been recently demonstrated that diet and nutritional status are among the most important modifiable determinants of human health, through a plethora of presumptive mechanisms among which microbiota-mediated processes are thought to have a relevant role. At present, probiotics and prebiotics represent a useful dietary approach for influencing the composition and activity of the human gut microbial community. The present study is composed of two main sections, aimed at elucidating the probiotic potential of the yeast strain K. marxianus B0399, as well as the promising putative prebiotic activity ascribable to four different flours, naturally enriched in dietary fibres content. Here, by in vitro studies we demonstrated that K. marxianus B0399 possesses a number of beneficial and strain-specific properties desirable for a microorganism considered for application as a probiotics. Successively, we investigated the impact of a novel probiotic yoghurt containing B. animalis subsp. lactis Bb12 and K. marxianus B0399 on the gut microbiota of a cohort of subjects suffering from IBS and enrolled in a in vivo clinical study. We demonstrated that beneficial effects described for the probiotic yoghurt were not associated to significant modifications of the human intestinal microbiota. Additionally, using a colonic model system we investigated the impact of different flours (wholegrain rye and wheat, chickpeas and lentils 50:50, and barley milled grains) on the intestinal microbiota composition and metabolomic output, combining molecular and cellular analysis with a NMR metabolomics approach. We demonstrated that each tested flour showed peculiar and positive modulations of the intestinal microbiota composition and its small molecule metabolome, thus supporting the utilisation of these ingredients in the development of a variety of potentially prebiotic food products aimed at improving human health.

The thesis deals with a theme of probiotic mikroorganisms which are aplicated as a dietary supplement. Health benefit on the host have been known for a long time, however, there might be some negative effects affecting a consumer. In connection with this observation defining requirements and evaluating of safety of probiotics in food was necessary. The new probiotic dietary supplement can not be expanded on the market without these processes. Every strain must be tested separately on each property. The ability of rezistance against conditions inside human gastrointestinal tract belongs to the most basic tests. The aim of the practical part is testing this ability of rezistance in vitro by simulated conditions. The model of gastrointestinal juices was designed according to the Czechoslovak codex. The form contributes to the higher rezistance of probiotics within implementing to the host. Also a dosage form of probiotics was involved to a in vitro testing.

Background Persistent diarrhoea (diarrhoea lasting more than 14 days) accounts for one third of all diarrhoea related deaths in developing countries in some studies. Probiotics may help treatment. Objectives To evaluate probiotics for treating persistent diarrhoea in children. Search methods We searched the Cochrane Infectious Diseases Group Specialized Register, CENTRAL, MEDLINE, EMBASE, and LILACS. We also contacted authors of included trials and organizations working in the field, and checked reference lists. The date of the most recent search was 13 December 2012 Selection criteria Randomized controlled trials comparing a specified probiotic agent with placebo or no probiotic in children with persistent diarrhoea. Data collection and analysis Two review authors assessed the eligibility, risk of bias, extracted and analysed data. Differences were resolved by discussion. Statistical analysis were performed using the fixed-effect model and the results were expressed as mean difference (MD) for continuous outcomes with 95% confidence intervals (CI). Main results Four trials were included, with a total number of 464 participants; one trial had a low risk of bias. Meta-analysis showed that probiotics reduced the duration of persistent diarrhoea (mean difference 4.02 days, 95%CI 4.61 to 3.43 days, n = 324, two trials). Stool frequency was reduced with probiotics in two trials. One trial reported a shorter hospital stay, which was significant, but numbers were small. No adverse events were reported.

O Brasil é o quinto maior produtor de leite do mundo, entretanto sua produtividade é considerada baixa. Um dos motivos desta baixa produtividade é a alta incidência da mastite no rebanho. A mastite é a reação inflamatória da glândula mamária, causada principalmente pela invasão e ação de espécies de micro-organismos patogênicos. O principal tratamento consiste no uso de antibióticos de aplicação intramamária. Entretanto, o uso constante de antibióticos apresenta problemas graves como o surgimento de cepas de bactérias resistentes e a presença de resíduos destes medicamentos no leite. Para reduzir esses problemas, formas alternativas de prevenção e tratamento da mastite têm sido estudadas, como o uso de preparações contendo espécies probióticas. Pesquisas recentes têm demonstrado o potencial de algumas destas espécies no tratamento e prevenção da mastite bovina tendo em vista sua atividade inibitória sobre os patógenos responsáveis por esta infecção. Desta forma, a proposta deste estudo foi desenvolver e avaliar a estabilidade de uma preparação probiótica na forma de emulsão de aplicação intramamária contendo 108 UFC/mL de um \"pool\" constituído de cinco cepas de Lactobacillus (L. acidophilus ATCC 4356, L. casei ATCC 7469, L. fermentum ATCC 9338, L. plantarum ATCC 8014 e L. paracasei Nestlé ST11). Para tanto, 12 formulações de emulsão base foram desenvolvidas e avaliadas em ensaios preliminares de estabilidade (centrifugação e estresse térmico), sendo a formulação contendo monestearato de glicerila, lanolina anidra, óleo mineral, Tween 80® e Spam 80® selecionada para formulação da preparação probiótica. Adicionalmente, selecionou-se como agente conservante sorbato de potássio visando aumentar a vida útil da preparação probiótica. A preparação, assim desenvolvida, foi submetida ao teste de estabilidade físico-químico por 90 dias a 50°C ± 2°C. Observou-se que o aspecto da preparação se manteve estável, mas com redução nos parâmetros pH, densidade e aumento na viscosidade. A manutenção da viabilidade dos Lactobacillus na preparação foi estudada em um ensaio de 120 dias, sendo avaliada a influência dos fatores: temperatura de estocagem, adição de sacarose e sorbato de potássio. Observou-se influência significativa da temperatura demonstrando a importância da refrigeração na manutenção na viabilidade dos Lactobacillus. Verificou-se também que, sob refrigeração, a adição de sacarose promoveu redução de 0,5 ciclo logarítmico em relação a população inicial de células viáveis. Observou-se redução do pH da preparação probiótica avaliada nestas condições. Estes resultados permitem concluir que a refrigeração da preparação é indispensável para a manutenção da viabilidade das bactérias probióticas. A adição de 2% de sacarose à preparação, mantida sob refrigeração, se mostrou eficiente na manutenção da população de células viáveis ao final de 120 dias de ensaio, contendo 108 UFC/g, característica desejável para um produto probiótico.
Brazil is the world\'s fifth largest milk producer, though its productivity has been considered low. One of the reasons for this low productivity is the high incidence of mastitis in the herd. Mastitis is the inflammation of the mammary gland, mainly caused by the invasion and action of pathogenic microorganisms. The treatment is accomplished through the use of intramammary application of antibiotics. However, the constant use of antibiotics has serious problems such as the emergence of resistant strains of bacteria and the presence of residues of these drugs in milk. To reduce these problems, alternative means of prevention and treatment of mastitis has been studied as the use of preparations containing probiotic species. Recent studies have demonstrated the potential of some of these species in the treatment and prevention of bovine mastitis due to their inhibitory activity on pathogen species responsible for the infection. Thus, the purpose of this study was to develop and evaluate the stability of probiotic preparation in the form of intramammary application emulsion containing 108 CFU/mL of five Lactobacillus strains (L. acidophilus ATCC 4356, L. casei ATCC 7469 L. fermentum ATCC 9338, ATCC 8014, L. plantarum and L. paracasei ST11 Nestle). Therefore, 12 basic emulsion formulations were developed and evaluated in preliminary tests for stability (thermal stress and centrifugation), and only one was selected. Additionally, potassium sorbate was selected as preservative in order to increase the shelf life of the preparation. The developed preparation was subjected to study the physicochemical stability for 90 days at 50 ° C ± 2 ° C. The results showed that the aspect of the preparation remained stable, but decreasing in pH and density and increasing in viscosity. The maintenance of the viability of the lactobacillus in the preparation was studied in a 120-day test, and flowing factors were considered: storage temperature, sucrose addition and the potassium sorbate addition. It was observed that there was significant differences in the temperature factor demonstrating the importance of cooling in maintaining the viability of Lactobacillus. It was also observed that sucrose decreased by 0,5 log cycle compared to initial population of viable cells in the refrigerated preparation. Regarding the pH, it was noted a decrease in the different formulations evaluated. These results show that the cooling of the preparation is indispensable for maintaining the viability of probiotic bacteria. The addition of 2% sucrose to the preparation proved efficient with a slower decrease in viability maintaining the population of viable cells at the end of the 120 days of the assay.

Background: HIV-1 infection results in marked immunologic insults, including pronounced CD4+ T-cell loss from gut lymphoid tissue, and structural damage to the intestinal mucosa following dysfunction of the gastrointestinal system, including compromised barrier function. Increased intestinal permeability and microbial translocation promote systemic immune activation, which is implicated in disease progression. While the development of highly active antiretroviral therapy (HAART) has been a major advancement in the treatment of HIV-1 infection, the need for novel complementary interventions to help repair intestinal structural and functional integrity remains unmet. Known properties of probiotics suggest that they may be useful tools in restoring normal intestinal flora. In this regard, the aim of the present study was to evaluate changes in microflora of the large intestine in chronic HIV infection and the possibility of correction by means of bacterial preparations (probiotics).

Post-operative infections are of high concern and are a great risk to patients who have just had an open-heart surgical procedure. Exposing the human body to elements it's clearly not used to, by opening the chest to perform heart surgery, greatly increases a patient's risk of infection. Surgery is usually the last line of defense used by physicians. This is not only because the invasiveness of the procedure, but because of the heightened risk of infection for the patient. In the treatment of surgical patients, the possibility of infection and protocol for treatment must be taken into account. Patients who undergo open-heart procedures, such as CABG's, Valve Repair/Replacement, Aneurysms, and Aortic Root Repair/Replacement, often have compromised immune systems. A compromised immune system, exposure of the chest cavity during surgery, and length of hospital stay are all contributing factors to hospital acquired infections (HAI). Infections vary depending on the nature of the surgery, the institution, surgical technique, and also varies by patient. Pneumonia, sepsis, bacterial endocarditis, and mediastinitis are all potential concerns in patients who have undergone recent open-heart surgery. The recent societal push for a more holistic approach to medicine and an effort for treatments with fewer side effects have lead to increased research in the realm of probiotics. Probiotics are "good bacteria" that are thought to aid in the intestinal microbial balance. They also have been shown to improve gastrointestinal problems. By integrating the use of probiotics into the treatment of post cardiac surgery patients, we expect to see a decrease in gastrointestinal problems and infections. We also believe that this will improve patients' overall health. This preventative treatment will save the hospital millions of dollars in revenue that is spent on treating infections and will also increase hospital bed turnover.

El papel de la dieta en el ser humano es el proporcionar suficientes nutrientes para satisfacer las necesidades metabólicas, además de brindar una sensación de satisfacción y bienestar. Hoy en día se sabe que los alimentos que consumimos juegan un papel importante en la modulación de diversas funciones fisiológicas de nuestro organismo. Las personas actualmente se preocupan más por los alimentos que consumen y sus beneficios a la salud que producen, por lo que gracias al avance del conocimiento sobre los que los alimentos pueden aportar, surgen los alimentos funcionales donde una de las categorías con mayor demanda y aceptación son los productos que contienen microorganismos beneficios a la salud, denominados alimentos con probióticos. La mayoría de los alimentos con probióticos se encuentran en el sector de lácteos, pero existe un segmento importante de la población que no los consume, por lo que el desarrollo de productos derivados de cereales con probióticos puede ser una alternativa para ellos. El objetivo de la investigación fue probar la factibilidad de la incorporación de microorganismos probióticos, específicamente Bacillus coagulans (Ganeden®) a una barra de cereal rellena de fruta y a una galleta de avena, y evaluar la sobrevivencia en condiciones de vida de anaquel. Se realizó una caracterización de la cepa de B. coagulans (Ganeden®) mediante pruebas: microbiológicas básicas, de funcionalidad, de inocuidad y de identificación molecular, posteriormente se analizaron los flujos de elaboración a nivel industrial de los productos para determinar la forma de incorporación de los microorganismos probióticos. Posterior al proceso tecnológico se evaluó en las diferentes fases de elaboración de los productos (ingredientes secos, masa, masa fermentada y producto terminado) la sobrevivencia del B. coagulans incorporado, La sobrevivencia en la vida de anaquel en seis meses almacenados a temperatura ambiente y bajo condiciones de abuso de temperatura también fueron evaluadas. En la presente investigación se comprobó la pureza, las características funcionales y de inocuidad del microorganismo utilizado B. coagulans (Ganeden®), así mismo se realizó una identificación con métodos moleculares hasta el nivel de especie, lo que da certeza del consumo en el caso de que el desarrollo llegara al mercado. De los productos a los que se les incorporó el microorganismo probiótico en esta investigación, la galleta de avena presentó mejor sobrevivencia, ya que no se observa reducción en el contenido de microorganismos en los 6 meses estudiados, mientras que la barra de cereal con relleno de fruta presentó una reducción de medio ciclo logarítmico en el mismo tiempo, por lo que se considera que es factible en un futuro el poder escalar de forma industrial el proceso estudiado y poner a disposición del consumidor una nueva opción de productos derivados de cereales con probióticos.
The role of diet in humans is to provide enough nutrients to satisfied metabolic needs, in addition to provide a sense of satisfaction and well-being. Today we know that the food we eat plays an important role in modulating physiological functions of our body. People nowadays are more concerned with the foods they eat and their health benefits, so thanks to the advance of the knowledge that food can provide functional foods arise where one of the categories with the highest demand and acceptance are products containing microorganisms with health benefits, called food with probiotics. Most probiotic foods are in the dairy sector, but there is a large segment of the population that does not consume them, so the development of probiotic-based cereal products may be an alternative for them. The objective of the research was to test the feasibility of incorporating probiotic microorganisms, specifically Bacillus coagulans (Ganeden®) into a fruit-filled cereal bar and an oatmeal cookie, and to evaluate survival in shelf-life conditions. A characterization of the B. coagulans strain (Ganeden ®) was carried out through basic microbiological, functional, safety and molecular identification tests, followed by an analysis of the industrial processing flows of the products to determine the way of incorporation of probiotic microorganisms. Subsequent to the technological process, the survival of B. coagulans was evaluated in the different stages of product processing (dry ingredients, dough, fermented dough and finished product). The survival during its shelf life during six months stored at room temperature and under conditions of temperature abuse were also evaluated. In the present investigation, the purity, functional and safety characteristics of the microorganism used B. coagulans (Ganeden®) were verified, as well as an identification with molecular methods up to the species level, which gives certainty of the consumption in the case this development reach to the market. Of the products to which the probiotic microorganism was incorporated in this research, the oat cracker presented better survival, since there was no reduction in the content of microorganisms in the 6 months studied, while the cereal bar filled with fruit presented a reduction of half log cycle at the same time, so it’s considered that in the future may be feasible to scale the process in an industrial way and to make available to the consumer a new choice of products derived from cereals with probiotics.

Ett av fem spädbarn drabbas av kolik under första tre levnadsmånaderna. I övrigt friska spädbarn skriker, gråter och är otröstbara minst tre timmar i sträck per dag mer än tre dagar i veckan. Trots att kolik är ett vanligt förekommande problem för både spädbarn och föräldrar finns det idag ingen effektiv behandling. Etiologin är oklar men rökning och mental ohälsa hos modern prepartum, födoämnesallergi och ett outvecklat magtarmsystem hos spädbarnet tros vara några bidragande faktorer. Denna litteraturstudie granskar åtta artiklar för att dels undersöka skillnaderna i tarmfloran mellan spädbarn med och utan kolik, om skillnaderna kan förklara kolik samt probiotikas effekt vid kolik. Granskade studier visar att förekomsten av Proteobacteria, koliforma bakterier och Escherichia coli är högre hos spädbarn med kolik. Till skillnad från spädbarn utan kolik uppvisar kolikbarn en mindre bakteriell mångfald, mindre kolonisation av Bacteroidetes och de gynnsamma bakteriesläkterna Lactobacillus och Bifidobacterium. Hos ammade kolikbarn som behandlades med den probiotiska bakteriestammen Lactobacillus reuteri DSM 17938 erhölls i två av tre studier  god effekt av probiotika utan några biverkningar där gråttiden reducerats till mer än hälften. Resultatet varierar geografiskt och evidensen kring probiotikas effekt hos flaskmatade spädbarn med kolik är fortfarande bristfällig. Det är oklart om en förändrad tarmflora kan orsaka kolik eller är ett resultat av koliken då tarmfloran är mycket komplex och fler studier inom området behövs.
One out of five infants suffer from colic during the first three months of life. Otherwise healthy infants suffer from severe crying and are unable to be comforted during at least three consecutive hours per day for more than three days a week. Although colic is a common problem for both babies and parents, there is currently no effective treatment. The etiology is unclear, but smoking and  prepartum maternal mental illness, allergies to foods and an undeveloped gastrointestinal system in infants are believed to be contributing factors. This study reviews eight articles that examine the differences in intestinal flora between infants with and without colic, to see if the differences can explain colic and probiotic effect in colicky infants. Reviewed studies show that the prevalence of Proteobacteria, coliforms and Escherichia coli is higher in infants with colic. Unlike infants without colic, colicky infants exhibit a minor bacterial diversity, reduced colonization of Bacteroidetes and the beneficial bacterial genus of Lactobacillus and Bifidobacterium. Breastfed colicky infants treated with the probiotic bacterial strain, Lactobacillus reuteri DSM 17938, obtained, in two out of three studies, a positive effect on colic symtoms without any side effects, where the crying time was reduced to less than half. Results vary geographically and evidence for a probiotic effect on bottle-fed infants with colic is still inadequate. It remains unclear whether an altered intestinal flora can cause colic or is a result of colic as the intestinal flora is very complex and more studies are needed in this area.

Bakgrund: Antibiotikabehandling kan medföra gastrointestinala besvär hos patienter. En vanlig orsak till detta är bakterien Clostridium difficile som normalt kan återfinnas i tarmfloran. Vid antibiotikabehandling kan flera av tarmflorans goda bakterier slås ut vilket kan resultera i att C. difficile växer till sig och genom toxiner ge gastrointestinala besvär. Detta kan orsaka patienter onödigt lidande och kostar hälso- och sjukvården mycket pengar årligen. Probiotika består av levande mikroorganismer och det finns teorier om att dessa kan ha en positiv inverkan på tarmfloran genom stabilisering av den samt förhindrar överväxt av C. difficile i floran.   Syfte: Syftet var att undersöka om behandling med probiotika kan förebygga C. difficile infektion och/eller lindra gastrointestinala besvär orsakade av C. difficile i samband med antibiotikabehandling.  Metod: Beskrivande litteraturöversikt. Litteratursökning gjordes i CINAHL och PubMed. Elva randomiserade kontrollerade studier utgjorde resultatet i litteraturöversikten efter att ha genomgått en kvalitetsgranskning. Resultat: En statistisk signifikant skillnad gällande förebyggande av C. difficile fanns i tre av de elva inkluderade studierna. Probiotika som symtomlindring mot C. difficile visade på en signifikant skillnad avseende diarré och lösa avföringar i fyra studier med signifikant fler fall i kontroll- än interventionsgrupperna. Andra symtom som undersöktes i sammanlagt sju studier var utspänd buk, buksmärta, illamående, kräkning och flatulens. Slutsats: Litteraturöversikten påvisade att probiotika i vissa fall kan ha effekt mot infektion orsakad av C. difficile i samband med antibiotikabehandling. Effekten visar sig i form av lindrande effekt på vissa gastrointestinala besvär. Litteraturöversikten kunde dock inte påvisa tillräcklig evidens på grund av varierande resultat i de inkluderade artiklarna. Det gör att probiotika i nuläget inte är aktuellt för implementering inom sjukvården i samband med antibiotikabehandling för att förebygga eller lindra infektion av C. difficile.
Background: Patients treated with antibiotics can experience gastrointestinal side effects. A common cause of the side effects is the bacterium Clostridium difficile. C. difficile can normally be found in the intestinal human flora. Antibiotic treatment can cause imbalance in the intestinal flora because antibiotics sometimes eliminate several of the good bacteria in the gastrointestinal microbiome. This can cause an overgrowth of C. difficile and through its toxins cause gastrointestinal side effects for the individual. The side effects of antibiotics and C. difficile can cause individual suffering and cost healthcare a lot of money annually. Probiotics contain living microorganisms and there are theories that these microorganisms can have a positive effect on the intestinal human flora by stabilizing the gastrointestinal microbiome and preventing overgrowth of C. difficile in the human flora.   Aim: The aim was to investigate if treatment with probiotics could prevent C. difficile infection and/or relieve the occurrence of gastrointestinal side effects caused by C. difficile in patients treated with antibiotics.    Method: A descriptive literature review. Data collection was made in CINAHL and PubMed. Eleven randomized controlled trials were included in this literature review after a quality review.   Results: A statistically significant difference in the prevention of C. difficile was found in three of the included articles. Probiotics used for relief of symptoms caused by C. difficile showed a significant difference in four studies that examined loose stools and diarrhea with more cases in the control group than the intervention group. Other symptoms examined in a total of seven articles were distended abdomen, abdominal pain, nausea, vomiting and flatulence.  Conclusion: This literature review proved that probiotics in some cases may have an effect on infections caused by C. difficile during treatment with antibiotics. The literature review could not prove sufficient evidence due to varying results in the included studies. That makes probiotics at the present time not relevant for implementation in healthcare services during treatment with antibiotics for prevention of C. difficile.

Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and PKU animals based on PAH allele inheritance. Ocular and developmental phenotypes in the PAHenu2 colony match those described in previous IGF-1 studies. Understanding the IGF-1 inheritance discrepancy will enable better osteopenia research using PAHenu2 mice and allow breeding of a healthier mouse colony for continued research. Collectively the results from this work describe a new therapeutic approach for treatment of PKU as well as a better understanding of the PAHenu2 mouse model to study this disease.

Probiotics are living microorganisms, which when administered in adequate amounts confer a health benefit on the host. They have been reported to relieve acute diarrhoea, atopic dermatitis and irritable bowel syndrome in disease-specific animal studies and in human intervention trials. However, probiotics are regularly consumed by general healthy population with limited knowledge in the immunomodulation of probiotics of local and systemic immune responses in healthy experimental models. Serving as the first line of defense against microbial infections and the largest immunological organ in animal host, the epithelium lining the small and large intestine is supposed to be the first organ to encounter probiotics as probiotics are always orally taken. It is believed that probiotics regulate the local immunities in the gut, which acts as the pivot in modulating the systemic immune responses. Accordingly, it was hypothesized that probiotic bacteria can modulate both local and systemic immune responses in healthy population; and the immunomodulation of combination of probiotics is different from that of individual strains. Wildtype healthy C57BL/6 mice were fed with different probiotic strains − Lactobacillus rhamnosus GG (LGG), Lactobacillus rhamnosus LC705 (LC705), Bifidobacterium breve Bb99 (Bb99), Propionibacterium freudenreichii ssp. shermanii JS (PJS) or Escherichia coli Nissle 1917 (EcN), or mixture of probiotics − GGmix (LGG, LC705, Bb99 and PJS) and ECPJSmix (PJS and EcN), for three weeks. After that, intestine, liver, spleen and blood were investigated. Probiotics suppressed intestinal T helper (Th)17 immune response but enhanced systemic (hepatic and splenic) Th17 immune response, suggesting that immune homeostasis was maintained in healthy individuals. Mechanism of action of LGG was further studied in this project as LGG is the widely studied probiotics. It was hypothesized that LGG exerts immunomodulatory effects by bacteria cells and/or its derived soluble factors such as lactic acid. Immunomodulatory effects of LGG cells and their soluble factors on dendritic cells (DCs), macrophages and monocytes from healthy blood donors were investigated as antigen-presenting cells (APCs) are pivots of bridging innate and adaptive immunities. Cytokine secretion profile, expressions of toll-like receptors (TLRs) and activation-related receptors of the APCs were examined. Both LGG cells and their soluble factors promoted type 1-responsiveness while soluble factors promoted type 17-responsiveness as well. Yet, lactic acid seemed not to be the one which enhanced type 1 and type 17 immune responses in soluble factors. With better understanding on the immunomodulation of probiotics in healthy models, prophylactic efficacy of probiotics in preventing infections and diseases can be availed.
published_or_final_version
Biological Sciences
Doctoral
Doctor of Philosophy

An imbalance of the T cell immune response is observed in inflammatory bowel disease. Intestinal microbes have been linked to the disease and the disease process leads to severe mucosal injury and systemic translocation of bacterial products. Aminosalicylates, corticosteroids and immunomodulators reduce these aggressive activities but are associated with potentially serious adverse events. The aim of this work was to investigate the effects of administration of prebiotics and probiotics that modulate the gut microflora and modulate the immune response, in ameliorating severity of colitis. The prebiotic, high amylose maize resistant starch was used at two different concentrations. A number of Bifidobacterium and Lactobacillus strains were used as probiotics. BALB/c mice were administered the prebiotics and probiotics and intrarectally infused with 2.5 mg trinitrobenzene sulfonic acid (TNBS) in 45% ethanol, thereby generating colitis. Mucosal cytokine responses, colonic microbial profiles and disease activity indices were monitored. The 5% concentration of high amylose maize resistant starch delayed progression of TNBS colitis as evidenced by reduced weight loss, lesser tissue damage, abrogation of the expression and synthesis of IFN-?? and upregulation of IL-4 and IL-10. The 30% concentration of high amylose maize resistant starch exacerbated the inflammatory response with an increase in acetic acid, coliforms and endopores in the colonic contents. Three strains of bifidobacteria and 3 strains of lactobacilli were individually screened for their activity against TNBS colitis. Each strain had a distinctive effect on the course of colon inflammation. Lactobacillus fermentum VRI 003 was selected for further study as it provided most protection. The ratio of immunosuppressive cytokines to pro-inflammatory cytokines was restored closer to the normal T cell cytokine levels. It also reduced the incidence of translocation of enteric bacteria into the spleens. Dosing a minimum daily dose of 6x109 CFU L. fermentum VRI-003 to ulcerative colitis patients in remission and maintained on standard therapy for 6 months prevented the exacerbation of symptoms, including diarrhea and abdominal pain, and improved the patient general well being. It also suppressed production of IFN-?? and sustained IL-10 levels. Moreover, absence of endospores and lower numbers of coliforms were detected in the faeces of UC patients during L. fermentum VRI-003 treatment. In summary, 5% high amylose maize resistant starch and L. fermentum VRI 003 prevented colon inflammation by changing the nature of the T cell immune response and modifying the colonic microflora in the murine model. The clinical evidence supported these findings.

Thesis (M.S.) University of Missouri-Columbia, 2006.
The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on August 21, 2007) Includes bibliographical references.

There is an increasing demand from the pharmaceutical and food industries for new probiotic products. Some yeast strains offer probiotic benefits, but it has not been possible to formulate them successfully into a tablet dosage form. Direct compression is regarded as the best technique for producing tablets containing dried yeast, however, the main challenge of tabletting a powder mixture containing yeast granules is the mechanical sensitivity of the cells. The objective of this work was to develop a rigid yeast tablet containing adequate numbers of viable yeast cells by optimizing the formulation to allow a relatively low compaction force to be used. In addition the mechanical properties of primary particles and granules were determined as an attempt to predict the compaction behaviour during tabletting. A tablet formulation was developed which exhibited tablets of an acceptable tensile strength. These tablets were further tested for friability, dissolution and storage stability to ensure it is fit for consumer use.

Probiotic bacteria have been investigated in the prevention and treatment of various diseases and allergies. The current study was undertaken to determine the effect of eight probiotic Lactobacillus species against bacterial human skin pathogens using several techniques. Antimicrobial activity of lactobacilli against Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Propionibacterium acnes was evaluated using lactobacilli broth cultures (BCB) and cell free supernatant (CFS). Antimicrobial activity was significantly greater with BCB compared with CFS especially for Lactobacillus plantarum and Lactobacillus acidophilus. Lactobacilli and pathogen aggregation, biofilm formation and adhesion to keratin were assessed. L. casei and L. plantarum were selected for further study as they showed the greatest co-aggregation (18.02 ± 1.34% with L. casei and 14.92 ± 1.45% with L. plantarum) with the pathogens (16.63 ± 1.65% with S. aureus 3761 and 14.58 ± 1.68% with P. aeruginosa) and prevention of biofilm formation by the pathogens. The antimicrobial activity of human beta defensin-2 (hBD-2) alone or with L. plantarum against pathogens was assessed. The results with hBD-2 showed that hBD-2 (10 μg / ml for 5 h) and L. plantarum together were significantly more inhibitory against S. aureus than hBD-2 alone. The presence of NaCl reduced the effectiveness of hBD-2 alone and with L. plantarum. In the presence of L. plantarum, inactivation of mprF and dlt genes led to increased binding of hBD-2 by the bacterial cell wall, and then inhibition growth of bacterial cell wall. Studies investigated the effect of exposure of methicillin resistant Staphylococcus aureus (MRSA) to the supernatant of L. plantarum the susceptibility of MRSA to β-lactams. MRSA became sensitive to β-lactams when treated with culture supernatant of L. plantarum. Gene expression studies demonstrated that the mecR1-mecI-mecA-PBP2 signalling pathway was impeded by exposure to culture supernatant of L. plantarum and β-lactams. The studies reported here demonstrate a possible alternative approach to dealing with skin pathogens, which may have clinical implications especially with regard to MRSA infections, and continued research is advised.

Includes bibliographical references (p. 119-130).
Production in farming of ostriches (Struthio camelus) is limited by the high mortality rate of ostrich chicks. Chicks which lack a well established microbiota are more susceptible to potentially fatal pathogenic infections. Therefore, the mortality rate may be decreased by establishing the correct gut microbiota by the use of ostrich specific probiotic strains. Five selected strains were conclusively identified and their mucin adhesion abilities characterised: Strain P1.2 was identified as Enterococcus faecalis; the identity of strain 5934.3.1 was confirmed to be Lactobacillus oris; Strains Lactobacillus brevis 512.3.1 and Lactobacillus oris 5934.3.1. The five selected strains were included in an in vivo probiotic feeding trial, where ostriches were treated with an encapsulated mixture of the five strains and/or the antibiotic tylosin.

Necrotising enterocolitis (NEC) is a serious gastrointestinal emergency that is associated with high mortality and morbidly especially in extremely low birth weight neonates. The burden of NEC is very high due to prolonged hospitalisation, and cost of surgery. Since the treatment is mainly supportive, prevention is the only option. Probiotics are defined as live microorganisms which when administered in adequate amounts confer a health benefit on the host. Systematic reviews published by our group (Deshpande et al, Lancet 2007 & Pediatrics 2010) before commencing this doctorate research have shown the potential of probiotics in prevention of NEC and mortality. Aims of this research included 1) establish the evidence-based guidelines to use probiotics 2) introduce routine use of probiotics and perform before and after cohort study 3) evaluate the effect of routine probiotics on gut and tracheal microbiome in very preterm neonates (<30 weeks gestation). 4) to advance this field further, i) perform systematic review of RCTs probiotics in low and medium income countries ii) evaluate the role of para-probiotics in preterm neonates. This thesis by publications contains 8 chapters. Chapter 3 is a publication about evidence-based guidelines to use probiotics in preterm neonates. Chapter 4 presents a stepwise approach to introduce probiotics and results of before and after cohort study reporting significant reduction of NEC in probiotic cohort. Chapter 5 discusses the clinical evidence of benefits of probiotics in preterm neonates along with guidelines for clinicians in the context of the Australian regulatory framework. Chapter 6 presents significant results of tracheal and gut colonisation by probiotic bacteria. Chapter 7 incorporates the systematic review results that suggest benefits of probiotics in low and medium income countries in terms of reduction of NEC, sepsis and mortality. Chapter 8 discusses the possible role of para-probiotics in preterm neonates. In summary, this research has added significantly to the existing body of evidence in the area of probiotics for prevention of NEC in high risk preterm neonates, contributed in advancing this area further, evaluated the role of probiotics in low and medium income countries and the role of para-probiotics in preterm neonates.

The objectives of this study were: 1) to investigate differentiation of Lactobacillus casei group by carbohydrate fermentation, PCR, gene sequencing and pulsed field gel electrophoresis, 2) to investigate tolerance of Lactobacillus strains in the presence of acid, bile salts and the or bile salt deconjugation, 3) to investigate viability of freeze dried Lactobacillus strains at various temperature during storage and their proteolytic activity, 4) to investigate the growth, viability and activity of Lactobacillus strains in skim milk containing prebiotics and 5) to investigate in vitro inhibition of Helicobacter pylori by Lactobacillus casei strains.

The objectives of this study were: 1) to investigate differentiation of Lactobacillus casei group by carbohydrate fermentation, PCR, gene sequencing and pulsed field gel electrophoresis, 2) to investigate tolerance of Lactobacillus strains in the presence of acid, bile salts and the or bile salt deconjugation, 3) to investigate viability of freeze dried Lactobacillus strains at various temperature during storage and their proteolytic activity, 4) to investigate the growth, viability and activity of Lactobacillus strains in skim milk containing prebiotics and 5) to investigate in vitro inhibition of Helicobacter pylori by Lactobacillus casei strains.

Il microbiota intestinale è riconosciuto come un fattore chiave per la salute umana, quindi la ricerca di strategie per migliorarne l'equilibrio è un campo di interesse emergente. I batteri probiotici possono stimolare il sistema immunitario e controllare la composizione del microbiota stesso. Invece, i prebiotici supportano i probiotici nell'ambiente intestinale. Infatti, un prebiotico non è digeribile dagli enzimi umani ma è disponibile per specifici batteri. Prebiotici e probiotici possono essere combinati insieme come synbiotici, in modo da migliorare la sopravvivenza e la crescita dei probiotici grazie ai prebiotici, per ottenere il massimo beneficio per la salute umana. Il progetto di dottorato ha come obiettivi la caratterizzazione di batteri probiotici che possano bilanciare il microbiota intestinale e migliorare la salute umana, e l’identificazione di nuovi prebiotici, testati attraverso modelli in vivo e in vitro. Verrà progettato almeno un simbiotico e valutati i suoi effetti benefici. I ceppi di Lactobacillus e Bifidobacterium sono stati caratterizzati per le loro proprietà probiotiche. Le loro capacità di fermentazione sono state testate su inuline commerciali e FOS differenti per il DP. I ceppi utilizzano preferibilmente FOS a catena corta e media come substrato di crescita. Inoltre, lo studio più approfondito di Lactobacillus plantarum PBS067 ha rivelato la capacità del ceppo di produrre una molecola batteriocina-like, con attività ad ampio spettro ed effetti sulle linee cellulari intestinali umane. La formulazione synbiotica progettata consiste in L. plantarum PBS067, Bifidobacterium animalis subsp. lactis PBS075 e Lactobacillus acidophilus PBS066 come probiotici, più i FOS adatti come prebiotici. La sua efficacia è stata testata su soggetti anziani sani assegnati in modo casuale a ricevere il prebiotico, il simbiotico o il placebo una volta al giorno per 28 giorni. La durata dello studio è stata di 56 giorni e all’inizio, dopo 28 giorni di somministrazione e dopo 28 giorni di follow-up, sono stati registrati la composizione del microbiota intestinale e i livelli di biomarcatori selezionati. Analizzando i profili tassonomici del microbiota, è stato evidenziato un arricchimento della comunità legato a un cambiamento significativo nei ceppi produttori di SCFAs dopo l'intervento simbiotico. I risultati sul benessere hanno suggerito l'efficacia del synbiotico nel ridurre l'incidenza dei sintomi CID legati ad una stimolazione positiva del sistema immunitario intestinale. Un estratto del fungo Grifola frondosa (Maitake) è stato valutato per il potenziale prebiotico. Le crescite dei probiotici, anche combinati in consorzio probiotico, erano paragonabili a quelle su FOS a basso DP, quindi il fungo è stato considerato un prebiotico. I metaboliti prodotti dopo la fermentazione di Maitake sono stati raccolti e analizzati tramite GC-MSD. La loro somministrazione a linee cellulari del colon-retto umane sane e cancerogene ha portato a una diminuzione della vitalità delle cellule tumorali e a un effetto protettivo dallo stress ossidativo. Infine, è stato sviluppato un microbiota intestinale ricostruito in vitro e convalidato imitando l’intervento synbiotico utilizzato in precedenza. La crescita dei batteri è stata monitorata come aumento di OD600nm e attraverso l'analisi dei metaboliti, mentre le modulazioni delle abbondanze dei singoli ceppi sono state valutate mediante qPCR. L'estratto di Maitake è stato usato come prebiotico al posto del FOS. I batteri più modulati sono risultati E. coli e L. plantarum e sono stati rilevati gli SCFAs.
The gut microbiota is recognized as a key factor in human health, so the search for strategies to improve its balance is an emerging field of interest. Probiotic bacteria could stimulate the immune system and control microbiota composition. Instead, prebiotics support probiotics in the intestinal environment. Indeed, a prebiotic is not digestible by human enzymes but is available for peculiar bacteria. Prebiotics and probiotics could be combined as synbiotics, expecting the enhancement of probiotic survival and growth thanks to prebiotics, to obtain the maximum benefit for human health. The PhD project aims to characterize probiotics balancing the gut microbiota and improving human health, and to identify new prebiotics, tested through in vivo and in vitro models. At least one synbiotic will be designed and its beneficial effects evaluated. Strains of Lactobacillus and Bifidobacterium were characterized for their probiotic properties. Their fermentation abilities were tested on commercial inulin and FOS differing for the DP. The strains preferentially used short- and medium-chain FOS as growth substrate. The deepest study of L. plantarum PBS067 revealed the ability to produce a bacteriocin-like molecule, with broad spectrum activity and effects on human intestinal cell lines. The synbiotic formulation designed consisted of Lactobacillus plantarum PBS067, Bifidobacterium animalis subsp. lactis PBS075 and Lactobacillus acidophilus PBS066 as probiotics, plus the suitable FOS as prebiotics. Its efficacy was tested on healthy elderly subjects randomly assigned to receive the prebiotic, the synbiotic, or the placebo once daily for 28 days. The study duration was 56 days and at the baseline, after 28 days of administration, and after 28 days of follow-up, the gut microbiota composition and the levels of selected biomarkers were recorded. Analysing the microbiota taxonomic profiles, an enrichment of the community linked to a significant change in the SCFAs producer strains was evidenced after the synbiotic intervention. The results on the wellbeing suggested the synbiotic effectiveness in reducing the CID symptoms incidence linked to a positive stimulation of the gut immune system. An extract from Grifola frondosa (Maitake) mushroom was assessed for the prebiotic potential. The growths of the probiotics, also combined as a probiotic consortium, were comparable to those on low DP FOS, so the mushroom was considered a prebiotic. The metabolites produced after Maitake fermentation were collected and analyzed via GC-MSD. Their administration to healthy and cancerogenic human colorectal cell lines led to a decrease in tumoral cell viability, and to a protective effect from oxidative stress. An in vitro reconstructed gut microbiota was developed and validated mimicking the previously used synbiotic. The growth was monitored as increase of OD600nm and through the analysis of the metabolites, while the modulations of single strain abundances were assessed through qPCR. Maitake extract was used as prebiotic instead of FOS. The most modulated bacteria were E. coli and L. plantarum and the SCFAs were detected.

Studi pregressi hanno mostrato che i micronutrienti come il boro (B) e il selenio (Se) e la vitamina D (VD) svolgono un ruolo chiave nel processo di ossificazione, tuttavia l’effetto delle interazioni fra queste diverse molecole, soprattutto a livello osseo, sono ancora poco conosciute. Inoltre, è nota la capacità dei probiotici di esercitare effetti benefici sull'organismo ospite. Lo scopo di questa tesi di dottorato è quindi quello di valutare l’effetto della somministrazione di B e Se e ceppi di probiotici sul processo di ossificazione e sul metabolismo osseo. I risultati ottenuti utilizzando specifiche linee reporter di osteoblasti di pesce zebra, la linea cellulare hFOB1.19 e l’integrazione delle evidenze ottenute con le analisi di trascrittomica hanno permesso di andare a fondo sui meccanismi attivati dalla sommistrazione dei diversi microelementi sul metabolismo osseo. Nelle larve di pesce zebra quello delle MAPK è risultato il pathway più regolato dalla co-somministrazione di B e VD, oltre alla segnalazione del TGF-β, all'adesione focale e alla segnalazione del calcio. Ciò ha suggerito che l’aggiunta di basse concentrazioni di micronutrienti alla dieta, può fornire un valido aiuto e ampliare le opzioni terapeutiche per il trattamento di condizioni patologiche causate da carenza di VD. Inoltre, il trattamento con i probiotici accelera la rigenerazione della pinna caudale nel pesce zebra, come suggerito dalla sovra-espressione di geni biomarker del sistema osseo e dall'aumento del contenuto dei fosfati misurato dall'analisi FTIRI a 5 giorni dopo l'amputazione (DPA) delle pinne. Inoltre, al fine di analizzare gli effetti di due ceppi probiotici, Bacillus subtilis e Lactococcus lactis, sulla formazione di osteoblasti e sulla mineralizzazione della matrice extracellulare, abbiamo creato due nuove linee transgeniche di pesce zebra contenenti sequenze codificanti proteine fluorescenti inserite nei geni che codificano per sp7 e col10a1a, in modo da poter marcare, rispettivamente lo sviluppo degli osteoblasti e della matrice extracellulare ossea. L’uso di queste linee transgeniche ci ha permesso di verificare che B.subtilis è stato il probiotico che ha permesso di recuperare al meglio recuperare gli effetti dannosi causati dall'inibitore del BMP. Inoltre, esperimenti sulle cellule hFOB1.19 hanno rivelato che l’esposizione all'estratto di B.subtilis ha aumentato significativamente i livelli di ALP. Inoltre, è stata anche confermata la validità dell’uso dei sistemi di coltura cellulare in vitro per testare gli effetti dei probiotici, utilizzando estratti batterici in alternativa alla somministrazione di cellule vive.
Micronutrients such as Boron (B) and Selenium (Se) are previously known to be playing key roles in ossification process but their interactions with VD is poorly explored. Probiotics are microbes known to exert beneficial effects on the host when supplied in adequate quantities. This research thesis aim to explore the osteogenic actions of B and Se, and selected probiotics strains. Specific zebrafish osteoblast reporter lines combined with transcriptomic analysis and hFOB1.19 cell line were used to explore the mechanism of actions. MAPK was confirmed as the most regulated pathway by the B and VD synergy groups in addition to TGF-β signaling, focal adhesion and calcium signaling in zebrafish larvae. By adding low concentrations of additional micronutrients, the results help broaden the therapeutic options for treating pathological conditions caused by VD deficiency. Probiotic treatment was found to accelerate the caudal fin regeneration in zebrafish confirmed by the upregulation of key marker genes and increased phosphates found by FTIRI analysis at 5 days post amputation (DPA) fins. Furthermore, to investigate the effects of two probiotics, Bacillus subtilis and Lactococcus lactis, on osteoblast formation and extracellular matrix mineralization, we created two new zebrafish transgenic lines containing fluorescent protein coding sequences inserted into endogenous genes for sp7 and col10a1a, that display fluorescence in developing osteoblasts and bone extracellular matrix, respectively. When larvae were exposed to a BMP inhibitor, B.subtilis was the most potent probiotics in recovering the bones from the detrimental effects caused by the BMP inhibitor. Additionally, experiments on hFOB1.19 cells revealed that B.subtilis extract significantly increased ALP levels via staining, western blot, and immunofluorescence. We also confirmed the suitability of in vitro cell culture systems to test probiotic effects by using probiotic extracts instead of live cells.

Pretenn birth is the most important cause of neonatal death and morbidity, with immense costs to families and health care systems. In spite of advances in obstetric care, approximately thirteen million pretenn birth occur annually world wide. Preterm birth is multifactorial in origin, but in the last 20 years, infection has emerged as an important and frequent cause of pretenn labour and birth. The most common pathway of infection into the uterus is the ascending route from the vagina. Since infection is associated with pretenn birth and there are adverse effects from antibiotic use, it is logical to ask whether antibiotics can prevent prematurity. Do antibiotics given systemically (for example: for urinary infection or dental caries) which have the potential to alter vaginal flora, decrease the incidence of pretenn birth? To assess the effectiveness of antibiotics given for any reason to prevent pretenn labour an umbrella review was carried out to assess systematically clinical trials of antibiotics and other relevant systematic reviews. Forty five clinical trials were included in the meta-analysis. The results indicated that the use of antibiotics during pregnancy for any reason had no effect in preventing pretenn labour before 34 weeks and this is accompanied by an increase in the maternal adverse effects. Another important finding was that metronidazole may increase the risk of pretenn labour. Antibiotics were also shown to be not effective in preventing neonatal morbidity associated with prematurity. A historical review was carried out as piece of methodological work to assess the numbers and quality of randomised clinical trials. The results confirmed our hypothesis that the number and quality of randomised clinical trials of antibiotics during pregnancy improved since the sixties. Probiotics may be a potential preventitive treatment for preterm labour. Normal vaginal flora is dominated by Lactobacilli which is associated with reduced risk of bacterial vaginosis and urinary tract infection. Probiotics consist of live Lactobacilli that are freeze dried with or with out sugar. Probiotic Lactobacilli displace bacteria, block pathogen attachment to vaginal epithelium and produce bacteriocins which inhibit pathogen multiplication. More importantly, lactobacilli produce hydrogen peroxide which maintains vaginal acidity - an unfavourable environment for pathogens. Probiotics have been shown to modulate the immune response by interfering with the inflammatory cascade that leads to preterm labour and birth. The administration of probiotics has been shown to be safe and effective in reducing and treating urogenital infections in non-pregnant populations. It is logical to ask whether probiotic therapy can restore the normal flora and may decrease the incidence of preterm birth? To assess probiotic's effectiveness and safety for preventing preterm labour and birth a Cochrane review was conducted. Four trials were assessed for inclusion in the review. One trial started in February 2005 and is still ongoing. One trial was excluded because there were no data to be extracted from the article. Of the two trials included in the review, one enrolled women after 34 weeks of pregnancy using oral fermented milk as probiotic, while the other study utilised commercially available yogurt to be used vaginally by women diagnosed with bacterial vaginosis in early pregnancy. Reduction in genital infection was the only prespecified clinical outcome for which the data were available. Results of this review showed that although the use of probiotics appears to treat vaginal infections in pregnancy, there are currently insufficient data from trials to assess impact on preterm birth and its complications. Given the lack of evidence highlighted in the Cochrane review, there was a compelling case to conduct a randomised controlled trial to test whether probiotics are effective in preventing preterm labour and birth or not. An exploratory trial was to be an important part of this thesis. A trial protocol was prepared but it was not approved by the MHRA. MHRA authorisation was refused because probiotics are licensed as food supplements and only medicinal products can be used in clinical trials. Given the failure to get MHRA approval, a decision was taken to perform an exploratory study to investigate if cervical immunology and morphology would better refme high risk status for future clinical trials. The hypothesis behind this cervix study is that a defective cervical barrier is a major cause of preterm labour. This hypothesis was assessed immunologically by using flow cytometry to quantity leucocytes subpopulation in the cervix, morphologically by cervical length and volume as assessed by 3D ultrasound, and the cervical blood circulation was assessed by 3D ultrasound with power Doppler in patients at high risk of preterm labour. Immunologically, the most prevalent cervical leukocyte was the macrophage. Macrophages and the rest of leukocyte types in general were fewer in high risk patients than in normal pregnant controls, but this difference did not reach statistical significance. This could be due to the sample size. Or this could be due to the collection of the cervical samples very early in pregnancy. The influx of leukocytes is expected later in pregnancy or at the time of cervical ripening prior to labour. Morphologically, this study showed a significant association between cervical length and preterm labour, but not cervical volume. This could be due to the sample size or due to the difficulty of estimating cervical volume. Also, there is increase in the vascularisation of the cervix in preterm labour patients and this could be due to the small sample size, difficulty of defining the cervix landmarks or premature remodelling of cervical architecture since all these women had previous histories of preterm labour.

Brachyspira are the causative agent of avian intestinal spirochaetosis, a gastrointestinal disease common in layer hens and broiler breeders. This disease costs the UK laying industry approximately £18 million per annum, resulting from reduced egg production and poor egg quality. Prevalence of Brachyspira is increasing, and due to the poor understanding of this pathogen, mitigation strategies have been largely unsuccessful. Therefore, preventative measures are essential. These studies aimed to improve the understanding of Brachyspira pathobiology and investigate Lactobacillus probiotics as a suitable mitigation strategy. Brachyspira and Lactobacillus species were characterised using phenotypic and genotypic methods. Four Lactobacillus isolates were selected for their inhibition of Brachyspira in vitro and demonstrated inhibition by a number of mechanisms. Secreted metabolites in Lactobacillus cell free supernatant inhibited Brachyspira (p value < 0.05) and metabolomic studies identified the production of organic acids to be a major contributor to inhibition. Protein denaturation in cell free supernatants significantly reduced Brachyspira inhibition (p value < 0.05), suggesting the role of bacteriocins in inhibition. Furthermore, L. reuteri isolates co-aggregated with Brachyspira in vitro, reducing pathogen viability (p value < 0.05). Pro-inflammatory responses to Brachyspira in HD11 avian macrophages were dominated by upregulation of IFNg (p value < 0.01) and pre-treatment of cells with Lactobacillus significantly reduced this response (p value < 0.0001), demonstrating the ability of probiotics to alter immune responses to Brachyspira. Galleria mellonella were utilised to study Brachyspira virulence and probiotic intervention. G. mellonella exhibited a varied response to Brachyspira iv infection and Lactobacillus isolates were able to protect against the mortality associated with Brachyspira isolates (p value < 0.05). The studies here demonstrated that Lactobacillus probiotics are a suitable mitigation strategy against Brachyspira. A number of mechanisms were identified, however future studies are required to explore these mechanisms in a more relevant in vivo chicken model.

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.

The equine cecal microbiome is an incredibly diverse ecosystem that is critical to the overall health of the horse. It is of particular interest to equine researchers because of the link between colic and the bacterial profile residing within the cecum. We investigated ten probiotics for their ability to reduce numbers of previously identified pathogenic microorganisms: Streptococcus bovis/equinus complex (SBEC), Escherichia coli K-12, Escherichia coli general, Clostridium difficile, and Clostridium perfringens. A preliminary in vitro study was used to measure the reduction in opportunistic bacteria that are commonly found in the equine gastrointestinal tract. The second in vitro was designed as a titration study using the three most effective probiotics from the first project. In this second stage of testing, different dosage levels were utilized to determine if dosage had an effect on bacterial reduction potential. Dosage levels included manufacturer’s recommended dosage (1x), twice the recommended dosage (2x), and three times the recommended dosage (3x). Lastly, an in vivo study was conducted using three cecally-cannulated horses in a Latin square design in order to measure opportunistic bacteria reduction potential in the live horse model. Cecal fluid characteristics, pH, volatile fatty acids, and ammonia concentrations were measured, along with bacterial concentrations. In the initial in vitro experiment, we observed that all probiotic treatments numerically decreased the bacterial concentrations in comparison to the control. There were three products that decreased bacterial concentrations most consistently: Command FT, CRS, and SmartDigest Ultra. These probiotics were chosen for a titration in vitro study. Selected probiotics were dosed at the recommended dose (1x), as well as at two times the recommended dose (2x) and three times the recommended dose. In the titration in vitro experiment, SmartDigest Ultra increased (P < 0.01) bacterial concentrations of E. coli K12 and C. perfringens at 2x and 3x. Due to a lack of statistical significance for Command FT and CRS (P ≥ 0.10), cost efficiency was used as a selection criteria. Command FT was approximately $0.27 per dose and CRS was approximately $0.18 per dose. Therefore, CRS was chosen for further testing in the following in vivo experiment. In the in vivo experiment, CRS was dosed at one times (1x) and two times (2x) the recommended dose. We observed that blood parameters, cecal fluid characteristics, and bacterial concentrations were not statistically (P ≥ 0.10) altered by treatment. Numerical increases for bacterial concentrations were observed for SBEC at 1x and 2x, C. difficile at 2x, and C. perfringens at 2x. Numerical decreases were observed for E. coli K12 at 1x and 2x and E. coli general at 1x and 2x. Overall, this study suggests that the selected probiotics can be safely used at the recommended dose.

The intestinal microflora can be considered as a postnatally aquired organ composed of a large diversity of bacterial cells that can perform different functions for the host. This organ is highly exposed to environmental influences and thus modulated in its composition and functions by external factors, such as nutrition. Specific bacterial components of the intestinal microflora, including lactobacilli and bifidobacteria, have been associated with beneficial effects on the host, such as promotion of gut maturation and integrity, antagonisms against pathogens and immune modulation. These two genera are mainly considered to include probiotic bacteria. Probiotics are live microorganisms which when administered in adequate amounts confer a health benefit on the host. This study has been done on probiotic strains investigating their different properties and activities. The antimicrobial and antifungal activities of two probiotic strains, Lactobacillus rhamnosus IMC 501® and Lactobacillus paracasei IMC 502®, and their 1:1 combination, named SYNBIO® were studied using four different methods. Using two modified streak methods and a well diffusion method, the inhibitory activity of the probiotics and their metabolites towards six Gram+, nine Gram- pathogenic bacterial strains and eight Candida strains, was tested. Antagonistic effect of probiotic Lactobacillus strains was also investigated by co-culturing assay highlighting a significant inhibition of most of the pathogens tested in this study. The combination SYNBIO® showed a microbicidal activity against most of the strains tested in the study. Compared to the control, most of the pathogenic bacteria and yeast were inhibited by all probiotic strains tested to various degrees. Screening Lactobacillus strains according to their activity in various environmental conditions could precede the clinical efficacy studies for adjunct treatment with probiotics in cure of different gastrointestinal and vaginal tract infections. There is also evidence suggesting that the consumption of fermented products containing lactobacilli and bifidobacteria may play a role in preventing cancer of the colon and rectum but the mechanisms responsible for this effect are not fully understood and may be linked to different factors. Colon cancer is a major cause of cancer death in affluent countries and diet plays an important role in its development. One of the essential components of the colonic cancer genesis is represented by DNA damage. Gut microbial products and food components are thought to be principally responsible for the damage that initiates disease progression. The two probiotic strains, L. rhamnosus IMC 501® and L. paracasei IMC 502®, and their combination, SYNBIO® were incubated at three different bacterial concentrations with a genotoxic human faecal water. HT29 cells were used to investigate the protective effects of the probiotic bacteria on DNA damage faecal water-induced by single cell gel electrophoresis (Comet) assay. The probiotic strains tested were not genotoxic and significantly decreased DNA damage induced by faecal water, indicating the potential to inhibit initiation. Incubation of faecal water with different concentrations of probiotics revealed that the decrease in genotoxicity was not dose dependent in some cases. In this study it was also investigated the influence of both probiotic strains on protein expression and MAPKs signalling pathways in HT29 cells, showing an improvement of the intestinal epithelium permeability. Consumers are more aware and concerned about their lifestyle than ever before. This has increased demand for foods that promote health and wellness, such as functional products containing probiotics and prebiotics, which have a beneficial effect on the balance of intestinal microbiota. Fermented dairy products are generally good food matrices to be enriched with probiotics, but the consumption of these products is limited due to growing vegetarianism and the large number of individuals who are lactose intolerant or on cholesterol-restricted diets. Thus, the development of non-dairy probiotic products, including food matrices based on fruit, vegetables and cereals, has been widely studied. This study reviews the main applications of probiotics in dairy and non-dairy food products and also in products of vegetable origin and the characteristics that enable the use of these food matrices as potential carriers of probiotic bacteria. However, in order to be beneficial, the bacterial cultures have to remain live and active at the time of consumption. One of the aim of this study was to develop new probiotic food products, such as seasoned cheeses, salami, chocolate and ice-cream with a final probiotic concentration of approximately 109 CFU/daily dose of L. rhamnosus IMC 501® and L. paracasei IMC 502® mixed 1:1 (SYNBIO®). The survival and viability of probiotics were determined during the foods shelf-life. The values of viable probiotic bacteria of all dairy and non-dairy foods were between 107 and 109 CFU/g of food at the end of the shelf-life and for some of them the values were maintained even after the expiry date. Based on the results of the current study, all the dairy (―Caciotta‖ cheese, ―Pecorino‖ cheese, ―Ba'¼scion‖ Swiss cheese and ―Fiordilatte‖ ice-cream) and non-dairy (―Ciauscolo‖ salami, Larded salami, Swiss small salami, milk chocolate, dark chocolate, organic jam and chocolate mousse) food products studied would be excellent vehicles to deliver the probiotic health effects because of the high viability of probiotics during the shelf-life of foods and in some cases even after their expiry date. The most well known food-based strategies to modulate the composition of the intestinal microbiota are the dietary use of prebiotics, probiotics and their combination, synbiotics. Since fermented foods have a great significance because they provide and preserve large quantities of nutritious foods in a wide diversity of flavours, aromas and texture, which enrich the human diet, a further aim of the study was to screen the effect of buckwheat flour and oat bran as prebiotics on the production of probiotic fibre-enriched fermented milks, by investigating the kinetics of acidification of buckwheat flour- and oat bran-supplemented milk fermented by L. rhamnosus IMC 501®, L. paracasei IMC 502® and their 1:1 combination, SYNBIO®. The probiotic strains viability, pH and sensory characteristics of the fermented fibre-enriched milk products, stored at 4°C for 28 days were also monitored. The results showed that supplementation of whole milk with the tested probiotic strains and the two vegetable substrates results in a significant faster lowering of the pH. Also, the stability of L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® during storage at 4°C for 28 days in buckwheat flour- and oat bran-supplemented samples was remarkable enhanced. The aim of this preliminary fermentation was to develop a new synbiotic product using the best combination of probiotics and prebiotics by promoting better growth and survival and be acceptable to the consumers with high concentration of probiotic strain. This new product was used to conduct a human feeding trial to validate the fermented milk as a carrier for transporting bacterial cells into the human gastrointestinal tract. The probiotic strains were recovered from faecal samples in 40 out of 40 volunteers fed for 4 weeks one portion per day of synbiotic fermented milk carrying about 10 9 viable cells. Probiotics have the potential to prevent and treat gastrointestinal and vaginal infections and diseases. This is an important field of further researches. Further well-conducted clinical studies using validated outcome measures are recommended. Among others, issues to be addressed are to further identify populations that would benefit most from probiotics/prebiotics administration, to determine the most effective dosing schedules, and to address the cost effectiveness of using probiotics and/or prebiotics for preventing and treating gastrointestinal and urovaginal infections. Undoubtedly, probiotic bacteria are most effective when safely and adequately integrated into one‘s diet. However, if used for therapeutic purposes, they should be ingested via capsules or tablets. It is important to remember that when used in pediatrics, as a preventive approach (e.g. in the case of acute diarrhea, antibiotic-induced diarrhea and allergy) their long-term use is more practical if the chosen bacteria are incorporated in the diet through yogurt, fermented milk, dairy and non-daity products or other beverages or foods consumed during the weaning process. Relative to daily supplement use, this approach helps with compliance and it reduces costs. The probiotic strains L. rhamnosus IMC 501® and L. paracasei IMC 502® are also used as deliverers in capsules or powders rather than in food form. Probiotic dietary supplements maintain for a long period of refrigerated and/or room temperature storage a high bacterial concentration, and can be sprinkled directly onto food; stirred into beverages, or taken as capsules. As the intestinal microbiota, also vaginal microbiota can change composition rapidly, for reasons that are not fully clear. This can lead to infection or to a state in which organisms with pathogenic potential coexist with other commensals. The most common urogenital infection in women is bacterial and fungal vaginosis. The majority of cases are caused by Candida albicans, but also C. glabrata, C. krusei and C. tropicalis can be problematic. In vitro studies have shown that Lactobacillus strains can distrupt bacterial vaginosis and yeast biofilms and inhibit the growth of urogenital pathogens. The use of probiotics to populate the vagina and prevent or treat infection has been considered for some time, but only quite recently have data emerged to show efficacy, including supplementation of antimicrobial treatment to improve cure rates and prevent recurrences. Therefore, we investigated the blockage of Candida pathogens adherence by Lactobacilli, under three possible mechanisms: exclusion by adhered Lactobacillus strains, displacement of adhered pathogens and competition for receptor sites (inhibition test). The inhibition results highlight a significant (P<0.05) competition of Lactobacillus plantarum 319, L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® against all the Candida strains. Since the SYNBIO® is a combination 1:1 of L. rhamnosus IMC 501® and L. paracasei IMC 502®, that are characterized as probiotics, it was used for the formulation of two new probiotic medical devices. Vaginal ovules and douches were produced using SYNBIO® as preliminary evaluation of different matrix and cell viability during 6 months. During the storage at room temperature for 6 months, Witepsol® was the matrix that showed the highest suitability to preserve viable microorganism thus increasing the shelf-life of the product respect to PEG formulation. The results suggest that the probiotic strains used in the present study could prevent colonization of the urogenital tract by relevant pathogens such as Candida strains through barrier and interference mechanisms (mainly exclusion and competition). An advantage for women is that they can self-administer the probiotics. Many more studies are needed to optimize the defensive properties of the vaginal microbiota, but the potential remains that the health of many women can be improved by probiotic intervention.

Darbo tikslas: ištirti probiotikų įtaką melžiamų karvių produktyvumui, produkcijos rodikliams ir sveikatingumui. Darbo uždaviniai: 1. Įvertinti probiotiko Probio - active įtaką melžiamų karvių pieno primilžiui ir pieno rodikliams. 2. Įvertinti probiotiko įtaką melžiamų karvių kraujo morfologiniams rodikliams. 3. Įvertinti probiotiko įtaką melžiamų karvių didžiojo prieskrandžio fermentaciniams procesams. Tyrimo metodika: tyrimas buvo atliekamas kooperatinėje bendrovėje „Blauzdžių agroservisas“, 2013 metų birželio – spalio mėnesiais. Bandymo metu buvo suformuotos dvi grupės melžiamų karvių: kontrolinė ir bandomoji, po 115 Lietuvos juodmargių veislės melžiamų 2-5 laktacijos karvių kiekvienoje. Kontrolinėje ir bandomojoje grupėse karvės buvo paskirstytos vienodai, atsižvelgiant į jų laktacijų skaičių ir veršiavimosi laiką. Bandomosios grupės karvės su įprastiniu racionu papildomai gavo probiotiko. Į kiekvienos bandomosios grupės karvių kombinuotuosius pašarus, kiekvieną šėrimo dieną individualiai, buvo įmaišoma po 150 ml probiotiko. Bandymas vyko nuo 60-os iki 120-os karvių laktacijos dienos ir truko 60 dienų. Pirmas 30 dienų bandymas vyko tvartiniu laikotarpiu, o paskutines 30 dienų – ganykliniu laikotarpiu . Bandymo metu buvo naudotas probiotikas Probio - active, įmaišytas į racioną normuojant po 150 ml kiekvienai karvei, normą padalinant į dvi dalis ir sušeriant paros laike per du šėrimus. Tyrimo metu mėginiai pieno tyrimams buvo imami bandymo pradžioje, praėjus 60 d., 90 ir... [toliau žr. visą tekstą]
Aim: to research the effects of probiotics on dairy cows’ productivity, productions values and the overall health. Objectives: 1. To evaluate the effects Probio – active probiotics toward dairy cows’ yield and milk index. 2. To evaluate the effects of probiotics on dairy cows morphological blood indexes. 3. To evaluate the effects of probiotics to the rumens fermentation processes on dairy cows. Methodology of the research: the research was carried out at the cooperative company “Blauzdžių Agroservisas” 2013 June - October. During the research two similar groups of dairy cows were created: the control group and experimental group. The groups were made out of 115 Lithuanian Black and White breed dairy cows in their 2-5 lactation stages each. Cows were considered and maintained under the same conditions. For the experimental group cows with normal diet each day in addition received 150 ml of a probiotic. The study samples of milk were taken for research at the beginning of the test after 60 days, 90 and 120 days, from the start. Haematological tests of dairy cows blood samples were taken at the beginning and the end. The rumen contents - at the beginning and after a two-month test. Results: during the research it was determined that the cows which received probiotics yielded 4.4 percent (p>0.05) more milk opposed to the ones that did not receive probiotics. It was also detriment that the use of probiotics have an effect on the quality indexes (protein, fat), and have... [to full text]

Bakgrund: Ett lämpligt näringsintag är viktigt för främjandet och bevarandet av både den allmänna och orala hälsan. Karies är en multifaktoriell sjukdom där kosten har en betydelsefull roll. Kosten kan bidra till både kariesförekomst och kariesprevention. Functional food definieras som ”livsmedel som ger hälsovinster utöver basnutritionen” och ”livsmedel som har en eller flera tillsatser som modifierats för att bidra till en hälsosam kosthållning”. Syfte: Syftet med studien var att kartlägga kunskapsnivån och tillämpningen av information till patienterna om Functional food i det kariesförebyggande arbetet inom Folktandvården, Landstinget Kronoberg. Metod: Enkätstudiens urval bestod av tandläkare, tandhygienister och tandsköterskor, totalt 106 deltagare, från 17 olika Folktandvårdskliniker tillhörande allmäntandvården i Landstinget Kronoberg. Enkäten bestod av 24 frågor som bedömdes enligt ett graderat poängsättningssystem. Resultatet redovisades deskriptivt med hjälp av tabeller och diagram. Statistiska analyser utfördes med hjälp chitvåtest. Resultat: Majoriteten av deltagarna kategoriserades i grupperna: låg kunskapsnivå samt låg till medelhög tillämpningsnivå av Functional food i det kariesförebyggande arbetet. Merparten av deltagarna hade ett stort intresse av att få ytterligare kunskap. Slutsats: Tandvårdspersonalen önskade mer kunskap om Functional food för en ökad tillämpning i patientarbetet, givet att vetenskaplig evidens uppnås.
Background: An adequate nutrition is essential for the promotion and maintenance of the general and oral health. Dental caries is a multifactorial disease in which diet is a important factor. Functional foods are defined as "foods that provide health benefits beyond basic nutrition" and "foods that has one or more additives, modified to contribute to a healthy diet". Aim: The aim of the study was to identify the level of knowledge and application regarding information to patients about Functional food in caries prevention work in Public dental services, County Council of Kronoberg. Method: The survey included 106 participants consisted of dentists, dental hygienists and dental nurses from 17 different Public dental services. The survey consisted of 24 questions. The results were presented descriptive, with tables and charts, as well as statistically analyzes using chi-square tests. Results: The majority of participants were categorized into the groups: low level of knowledge and medium or low application level of Functional food in the caries prevention work. Most of the participants had a great interest in obtaining additional knowledge. Conclusion: Dental professionals would like to get more knowledge about Functional food for an increased application in patient care, provided that scientific evidence is obtained.

This diploma thesis deals with encapsulation of natural extracts. In the theoretical part the methods of encapsulation, materials for particle preparation, as well as application of encapsulation techniques in food industry were described. In experimental part selected natural extracts of propolis, green barley and probiotics were characterized. There substances were encapsulated into alginate and chitosan. In the total of 25 types of prepared particles long-term stability in some model physiological conditions as well as in four different model foods was evaluated. Additionally, stability of selected particles in several real milk-based products was followed too. The stability of particles was determined spectroptohometrically. In natural extract a content of polyphenols, proteins, chlorophylls, as well as total antioxidant activity were analysed. To analysis of probiotics optical and fluorescence microscopy were used. In propolis and green barley antimicrobial activity was tested too. Moreover, in the sample of propolis also cytotoxic assay was applied. Agar-chitosan was chosen as the best shell material for propolis due to its optimal stability in model physiological conditions as well as model foods. Liposomes were evaluated as unstable and were not recommended for further application. As the suitable shell material for powdered green barley starch-alginate (rate 1:4) and agar-chitosan were proposed, while the second one showed better stability for released proteins. Agar-chitosan shell material was usable for fresh green barley too. For probiotics encapsulation alginate or alginate-starch were chosen because of their porosity and possibility of nutrients diffusion. In real foods the best results were reached with application of probiotic particles into milk. Coencapsulation of powdered barley and probiotics did not confirm inhibition of culture growth. Neither the antimicrobial effect of propolis and barley nor the cytotoxic effect of propolis were confirmed.

Trabalho Final de Mestrado Integrado, Ciências Farmacêuticas, Universidade de Lisboa, Faculdade de Farmácia, 2019
O conhecimento sobre a interação entre microorganismos e humanos evoluiu muito desde os dias em que todos estes seres eram considerados perigosos para a saúde. Atualmente, a microbiota é reconhecida como uma parte única e importante do nosso organismo e, o seu estudo contínuo ao longo dos anos, tem possibilitado novas oportunidades a produtos que consigam de alguma forma influenciá-la. Neste contexto, os probióticos são um bom exemplo desses produtos. Os probióticos são definidos como “microorganismos vivos que quando administrados em quantidades adequadas conferem benefício à saúde do hospedeiro”, e desde que foram primeiramente descritos em 1965, o interesse e uso como produtos promotores de saúde tem crescido, bem como os benefícios propostos sendo os géneros dos Lactobacillus e Bifidobacterium os mais usados. Embora estes produtos sejam considerados por muitos como produtos sem riscos para a saúde, é pertinente colocar-se a questão: serão os probióticos seguros? Os benefícios destes produtos são numerosos e estudados por vários autores. Os mais conhecidos encontram-se ao nível dos sistemas digestivo e imunitário. A prevenção e/ou tratamento adjuvante em neoplasias, a melhoria de estados depressivos e a redução dos níveis de colesterol são outros benefícios descritos. No entanto, tal como outros produtos na área da saúde, têm uma vertente de riscos e reações adversas, que só agora merece mais atenção por parte da comunidade científica. Algumas das preocupações de segurança descritas incluem a resistência destes microorganismos a antibióticos e a possível transferência dessa resistência a outras espécies, infeções bacterianas/fúngicas ou alterações farmacocinéticas. Para além dos não tão falados riscos, existem outras limitações e constrangimentos na sua utilização tais como a sua regulação e controlo pois não têm nenhuma legislação única e obrigatória associada. Alguns são considerados medicamentos, assegurando a sua segurança, qualidade e eficácia, mas na grande maioria dos casos são classificados como suplementos alimentares onde o controlo é menos apertado. O trabalho aqui apresentado é uma revisão de vários tópicos relacionados com os probióticos de maneira a melhor compreender o universo em que se insere, com o objetivo principal de descrever e reunir os menos conhecidos efeitos secundários e riscos associados ao seu consumo.
The knowledge about the interaction between microorganisms and humans has come a long way since the days where all these beings were considered dangerous for human health. Nowadays, our microbiota is being acknowledged as an important and unique part of numerous of our body sites and functions and its continued study has open new opportunities to products that somehow can influence and regulate it. In this context, probiotics are a good example of such products. Probiotics are defined as “live microorganisms which when administered in adequate amounts confer a health benefit on the host”, and since they were first described in 1965, their interest and usage as health promoter agents has been growing, as well as the proposed benefits, being the Lactobacillus and Bifidobacterium genera the most used. Although these products are overall considered by many to be risk free, is becomes necessary to ask: Are probiotics safe? The benefits of these products are many. The better well-known are in the digestive and immune systems but there are others that deserve to be mentioned. Prevention and/or adjuvant treatment of cancer, amelioration of depressive symptoms and reduction of cholesterol levels are some of them. Just like many other health connected products, probiotics have risks and side effects associated with them that weren’t a concern for a very long time but have now been gaining attention by the scientific community. Some of the safety concerns described include antibiotic resistance of these organisms and their possibility of transferring those genes to other bacterial species, bacterial and fungal infections or alteration of pharmacokinetics. Beyond the not so spoken risks, there are other limitations and constrains in their use, namely in their regulation and control as there isn’t a unique, mandatory legislation associated. Some are classified as drugs, with their safety, quality and efficacy assured whereas the great majority are regarded as supplements being their control less restrict. The present work is a review of several subjects regarding probiotics to better understand today’s probiotic universe with the main objective of describing and collecting the not so well known side effects and risks of probiotic consumption.