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Foram conduzidos três experimentos no confinamento experimental do Departamento de Zootecnia da ESALQ/USP com o objetivo de estudar os efeitos de diferentes aditivos em rações para bovinos terminados em confinamento. No experimento 1 foram utilizados 100 bovinos machos Nelore castrados (392 kg), distribuídos em 20 baias, por 60 dias. As rações continham 41% de sorgo moído, 40% de polpa cítrica peletizada e 15% de silagem de cana-de-açúcar. Os tratamentos foram: (1) controle, (2) monesina sódica Rumensin (MON1), (3) monensina sódica Rumenfort (MON2), levedura Saccharomyces cerevisiae Yea-Sacc 1026 (LEV1) e levedura Saccharomyces cerevisiae Biosaf SC47 (LEV2). A IMS foi reduzida pelo tratamento MON1 (P<0,05), quando comparada ao tratamento controle. O GPD dos animais não foi afetado pelos tratamentos (P>0,05). A EA dos animais não foi afetada pelos tratamentos (P>0,05). O tratamento MON2 apresentou um menor rendimento de carcaça (P<0,05) e o tratamento MON 1 apresentou uma maior AOL (P<0,05). No experimento 2 foram utilizados 96 tourinhos Nelore não castrados (396 kg), distribuídos em 16 baias por 95 dias. Os tratamentos foram: (1) Controle, (2) levedura Saccharomyces cerevisiae Biosaf SC47 (LEV), (3) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 1g /bovino/dia (PROB1) e (4) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 3g/bovino/dia (PROB2). As rações continham 59% de polpa cítrica peletizada, 35% de farelo de glúten de milho úmido e 5% de feno de tifton 65. A adição dos aditivos levedura (Saccharomyces cerevisae) e a combinação de leveduras e bactérias probióticas não afetou a IMS o GPD e a EA dos animais (P>0,05). A energia líquida de manutenção e de ganho das rações também não foi afetada pelos tratamentos (P>0,05), assim como os dados de carcaça. No experimento 3, que avaliou a digestibilidade das rações, foram utilizados 20 tourinhos Nelore, alocados em 20 baias individuais durante 15 dias, sendo 10 dias de adaptação ao marcador e 5 dias de coleta de fezes. A ração foi a mesma utilizada no experimento 2, e o marcador utilizado foi o óxido de cromo. Os tramentos utilizados foram: (1) Controle, (2) monensina sódica Rumenfort (MON), (3) levedura Saccharomyces cerevisiae Biosaf SC47 (LEV), (4) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 1g /bovino/dia (PROB1) e (5) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 3g/bovino/dia (PROB2). Os aditivos testados não afetaram as digestibilidades da MS, da MO, da FDN e da PB das rações (P>0,05). Com os resultados obtidos é possível afirmar que bovinos Nelore, castrados ou não, confinados com rações com altos teores de concentrado ricos em co-produtos como polpa cítrica e farelo de glúten de milho úmido não apresentam melhor desempenho nem melhor digestibilidade dos nutrientes quando suplementados com monensina sódica ou com micorganismos probióticos. / Three trials were conducted at the ESALQ/USP Animal Sciences Department experimental feedlot to evaluate the effects of different feed additives in feedlot finished cattle. On trial 1 100 Nellore steers (392kg) were allocated to 20 pens for 60 days. Experimental rations had 41% ground milo, 40% dried citrus pulp and 15% sugarcane silage. Treatments were: (1) control, (2) sodium monensin Rumensin (MON1), (3) sodium monensin Rumenfort (MON2), (4) yeast culture Saccharomyces cerevisiae Yea-Sacc 1026 (LEV1) and (5) yeast culture Saccharomyces cerevisiae Biosaf SC47 (LEV2). DMI was reduced by MON1 trestment (P<0,05) in relation to control. WDG was not affected by treatments (P>0,05). Treatments did not affect FE (P>0,05). Animals on treatment MON2 showed the lowest dressing percentage (DP) and those on MON1 showed the highest rib eye area (REA) (P<0,05). Trial 2 used 96 young Nellore bulls (396kg), allocated to 16 pens for 95 days. Treatments were: (1) control, (2) Saccharomyces cerevisiae Biosaf SC47 (LEV), (3) Saccharomyces cerevisiae and probiotic bacteria mix at 1g /animal/day dose (PROB1) and (4) Saccharomyces cerevisiae probiotic bacteria mix at 3g/animal/day dose (PROB2). Rations contained 59% dried citrus pulp, 35% wet corn gluten feed and 5% Tifton 65 hay. Treatments didnt affect DMI, WDG and FE (P>0,05). Rations net energy for maintenance and gain were also not affect by treatments (P>0,05), well as carcass data. Trial 3 utilized 20 young Nellore bulls allocated to individual pens for 15 days (10 days for adaptation to marker and 5 days for data collection) to evaluate rations digestibility. Experimental ration was the same utilized on trial 2, with chromium oxide as an external marker. Treatments were (1) control, (2) sodium monensin Rumenfort (MON1), (3) yeast culture Saccharomyces cerevisiae Biosaf SC47 (LEV), (4) Saccharomyces cerevisiae and probiotic bacteria mix at 1g /animal/day dose (PROB1) and (5) Saccharomyces cerevisiae probiotic bacteria mix at 3g/animal/day dose (PROB2). Treatments did not affect rations DM, OM, NDF and CP digestibilities (P>0,05). Results show that Nellore cattle, castrated or not, feedlot finished receiving rations with high levels of byproducts such as dried citrus pulp and wet corn gluten feed dont have higher performance nor better nutrients digestibility when supplemented with sodium monensin or probiotic microorganisms.

Made available in DSpace on 2016-06-02T19:02:44Z (GMT). No. of bitstreams: 1 6426.pdf: 2868015 bytes, checksum: e3bf13cc85e7606dc2d0f7dbb387ebaa (MD5) Previous issue date: 2014-11-24 / Financiadora de Estudos e Projetos / The evaluation of new materials and biomaterials modified with silver nanoparticles is a line of research that has been gaining strength in various knowledge areas including dentistry. Even considering international scientific publications, however, there are still few studies evaluating dental restoration materials that are modified with silver nanoparticles. Proposal: this study aimed to compare a photopolymerizable composite resin modified with an antimicrobial silver nanoparticle additive (an experimental resin), with the photopolymerizable composite resin in its unmodified form (a commercially available dental restoration material), by evaluating the in vitro antibacterial properties and also some of the primary physical properties of the resin. Methodology: composite resin disks were made for three experimental groups: unmodified resin (control group) and modified resin with different silver nanoparticle concentrations: 0.3% and 0.6% wt (groups 1 and 2). The antibacterial activity on the surface of the samples was evaluated by in vitro growth of of Streptococcus mutans and Lactobacillus acidophilus biofilms in a 20% sucrose medium, which is followed by counting the viable cells from the biofilms through serial dilutions after three incubation periods - 1, 4 and 7 days (n = 9). Optical transmittance was tested to measure the percentage of transmittance in the samples (n = 9). The surface roughness was evaluated using Atomic Force Microscopy (n = 9). The colour of the samples and the colour difference between the groups was measured in a colorimetry test according to the CIELab system. The presence of silver in the nanoparticle additive was analysed using X-Ray Fluorescence Spectroscopy. The quality of the dispersion and distribution of silver on the samples was analysed using Transmission Microscopy. The liberation of silver in artificial saliva was also evaluated in vitro after three incubation periods (60, 120 and 210 days) with agitation of the samples, using the Inductively Coupled Plasma Optical Emission Spectroscopy technique. Results: for the three incubation times, the number of viable cells was statistically lower in groups 1 and 2 than in the control group (p < 0.05), and groups 1 and 2 were not statistically different (p > 0.05) for the two bacterial species (ANOVA Two way/Tukey). There was no significant difference in optical transmittance between group 1 and the control (p > 0.05), and group 2 had a lower transmittance value than the other groups (p < 0.05) (ANOVA/Tukey). The analysis of the values of the arithmetic roughness, which was obtained using the NanoScope software, showed that there was no significant difference in surface roughness between the three groups (p > 0.05) (ANOVA/Tukey). From the measurements of the colours, the total colour difference (&#916;E) between all of the groups was higher than 1 (the minimum value that represents a colour alteration perceptible to human vision) and lower than 3,3 (critical &#916;E). The presence of silver in the nanoparticle additive was confirmed in the X-Ray Fluorescence test. Microscopic analysis showed good distribution of silver for groups 1 and 2 (with silver in the four quadrants of the regions analysed). The dispersion was better for group 1 than for group 2 (which showed a higher number of particle clusters). After the three incubation times, silver was not detected in the artificial saliva for the three groups or; if it was present, the concentration was below the limit of detection for this technique (< 0.02 mg/L). Conclusions: the antibacterial property on the surfaces of the experimental resins was verified for the two cariogenic bacteria considered. Furthermore, the addition of silver did not cause negative alterations to the resin modified with the lower concentration of nanoparticles, which makes the continuation of this study possible and valuable. / A avaliação de novos materiais e biomateriais modificados com nanopartículas de prata é uma linha de pesquisa que vem ganhando força em várias áreas do conhecimento, inclusive na área odontológica. Porém, mesmo considerando a publicação científica internacional, ainda há poucas pesquisas que avaliam materiais restauradores dentários modificados desta forma. Proposição: o propósito deste estudo foi comparar a resina composta fotopolimerizável modificada com um aditivo antimicrobiano nanoparticulado de prata (uma resina experimental), com a resina composta fotopolimerizável em sua forma não modificada, um material restaurador dentário disponível comercialmente, avaliando a propriedade antibacteriana, in vitro, e também algumas de suas principais propriedades físicas. Metodologia: foram confeccionados discos de resina composta para 3 grupos experimentais: resina não modificada (grupo controle), e resina modificada com diferentes concentrações de nanopartículas de prata, 0,3 e 0,6% (grupos 1 e 2). A atividade antibacteriana na superfície das amostras foi avaliada pelo crescimento in vitro de biofilme de Streptococcus mutans e Lactobacillus acidophilus em meio de cultura contendo 20% de sacarose, seguido de contagem de células viáveis provenientes dos biofilmes após 3 períodos de incubação, 1, 4 e 7 dias, através de diluições seriadas (n=9). Um ensaio de transmitância óptica foi realizado para medir a porcentagem de transmitância das amostras (n=9). A rugosidade superficial foi avaliada por Microscopia de Força Atômica (n=9). A cor das amostras e a diferença de cor entre os grupos foi medida em um ensaio de colorimetria considerando o sistema CIELab. A presença de prata no aditivo nanoparticulado foi analisada por meio da Espectrometria de Fluorescência de Raios X. A qualidade da dispersão e distribuição da prata nas amostras foi analisada através da Microscopia de Transmissão. Foi também avaliada a liberação de prata em saliva artificial, in vitro, após 3 períodos de incubação das amostras, sob agitação, 60, 120 e 210 dias, através da técnica de Espectrometria de Emissão Óptica por Plasma Acoplado Indutivamente. Resultados: para os 3 tempos de incubação, o número de células viáveis foi estatisticamente mais baixo nos grupos 1 e 2 do que no grupo controle (p < 0,05), e os grupos 1 e 2 não mostraram entre si diferença estatisticamente significante (p > 0,05), para as 2 espécies bacterianas (ANOVA Two way/Tukey). Não houve diferença significante na transmitância óptica entre o grupo 1 e o controle (p > 0,05), já o grupo 2 apresentou um valor de transmitância menor que os demais grupos (p < 0,05) (ANOVA/Tukey). A análise dos valores de rugosidade aritmética, obtidos através do software NanoScope, mostrou que não houve diferença significante na rugosidade superficial entre os 3 grupos (p > 0,05) (ANOVA/Tukey). A partir das medições de cores, a diferença total de cor (&#916;E) entre todos os grupos foi superior a 1 (valor mínimo que representa uma alteração de cor perceptível à visão humana) e inferior a 3,3 (&#916;E crítico). A presença de prata no aditivo nanoparticulado foi confirmada no ensaio de Fluorescência de Raios X. A análise microscópica demonstrou boa distribuição de prata para o grupo 1 e 2 (com prata nos 4 quadrantes das regiões analisadas), e a dispersão foi melhor para o grupo 1 do que para o grupo 2 (que apresentou maior número de aglomerados de partículas). Após os 3 tempos de incubação, não foi detectada a presença de prata na saliva artificial, para os 3 grupos, sendo que, se presente, ela encontrava-se em uma concentração abaixo do limite de detecção da técnica (< 0,02 mg/L). Conclusões: a propriedade antibacteriana foi verificada nas superfícies das resinas experimentais para as duas espécies bacterianas cariogênicas consideradas, e a adição de prata não provocou alterações negativas na resina modificada com a menor concentração de nanopartículas, o que viabiliza a continuidade do estudo.

Foram conduzidos três experimentos no confinamento experimental do Departamento de Zootecnia da ESALQ/USP com o objetivo de estudar os efeitos de diferentes aditivos em rações para bovinos terminados em confinamento. No experimento 1 foram utilizados 100 bovinos machos Nelore castrados (392 kg), distribuídos em 20 baias, por 60 dias. As rações continham 41% de sorgo moído, 40% de polpa cítrica peletizada e 15% de silagem de cana-de-açúcar. Os tratamentos foram: (1) controle, (2) monesina sódica Rumensin (MON1), (3) monensina sódica Rumenfort (MON2), levedura Saccharomyces cerevisiae Yea-Sacc 1026 (LEV1) e levedura Saccharomyces cerevisiae Biosaf SC47 (LEV2). A IMS foi reduzida pelo tratamento MON1 (P<0,05), quando comparada ao tratamento controle. O GPD dos animais não foi afetado pelos tratamentos (P>0,05). A EA dos animais não foi afetada pelos tratamentos (P>0,05). O tratamento MON2 apresentou um menor rendimento de carcaça (P<0,05) e o tratamento MON 1 apresentou uma maior AOL (P<0,05). No experimento 2 foram utilizados 96 tourinhos Nelore não castrados (396 kg), distribuídos em 16 baias por 95 dias. Os tratamentos foram: (1) Controle, (2) levedura Saccharomyces cerevisiae Biosaf SC47 (LEV), (3) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 1g /bovino/dia (PROB1) e (4) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 3g/bovino/dia (PROB2). As rações continham 59% de polpa cítrica peletizada, 35% de farelo de glúten de milho úmido e 5% de feno de tifton 65. A adição dos aditivos levedura (Saccharomyces cerevisae) e a combinação de leveduras e bactérias probióticas não afetou a IMS o GPD e a EA dos animais (P>0,05). A energia líquida de manutenção e de ganho das rações também não foi afetada pelos tratamentos (P>0,05), assim como os dados de carcaça. No experimento 3, que avaliou a digestibilidade das rações, foram utilizados 20 tourinhos Nelore, alocados em 20 baias individuais durante 15 dias, sendo 10 dias de adaptação ao marcador e 5 dias de coleta de fezes. A ração foi a mesma utilizada no experimento 2, e o marcador utilizado foi o óxido de cromo. Os tramentos utilizados foram: (1) Controle, (2) monensina sódica Rumenfort (MON), (3) levedura Saccharomyces cerevisiae Biosaf SC47 (LEV), (4) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 1g /bovino/dia (PROB1) e (5) combinação de levedura Saccharomyces cerevisiae e bactérias probióticas na dose de 3g/bovino/dia (PROB2). Os aditivos testados não afetaram as digestibilidades da MS, da MO, da FDN e da PB das rações (P>0,05). Com os resultados obtidos é possível afirmar que bovinos Nelore, castrados ou não, confinados com rações com altos teores de concentrado ricos em co-produtos como polpa cítrica e farelo de glúten de milho úmido não apresentam melhor desempenho nem melhor digestibilidade dos nutrientes quando suplementados com monensina sódica ou com micorganismos probióticos. / Three trials were conducted at the ESALQ/USP Animal Sciences Department experimental feedlot to evaluate the effects of different feed additives in feedlot finished cattle. On trial 1 100 Nellore steers (392kg) were allocated to 20 pens for 60 days. Experimental rations had 41% ground milo, 40% dried citrus pulp and 15% sugarcane silage. Treatments were: (1) control, (2) sodium monensin Rumensin (MON1), (3) sodium monensin Rumenfort (MON2), (4) yeast culture Saccharomyces cerevisiae Yea-Sacc 1026 (LEV1) and (5) yeast culture Saccharomyces cerevisiae Biosaf SC47 (LEV2). DMI was reduced by MON1 trestment (P<0,05) in relation to control. WDG was not affected by treatments (P>0,05). Treatments did not affect FE (P>0,05). Animals on treatment MON2 showed the lowest dressing percentage (DP) and those on MON1 showed the highest rib eye area (REA) (P<0,05). Trial 2 used 96 young Nellore bulls (396kg), allocated to 16 pens for 95 days. Treatments were: (1) control, (2) Saccharomyces cerevisiae Biosaf SC47 (LEV), (3) Saccharomyces cerevisiae and probiotic bacteria mix at 1g /animal/day dose (PROB1) and (4) Saccharomyces cerevisiae probiotic bacteria mix at 3g/animal/day dose (PROB2). Rations contained 59% dried citrus pulp, 35% wet corn gluten feed and 5% Tifton 65 hay. Treatments didnt affect DMI, WDG and FE (P>0,05). Rations net energy for maintenance and gain were also not affect by treatments (P>0,05), well as carcass data. Trial 3 utilized 20 young Nellore bulls allocated to individual pens for 15 days (10 days for adaptation to marker and 5 days for data collection) to evaluate rations digestibility. Experimental ration was the same utilized on trial 2, with chromium oxide as an external marker. Treatments were (1) control, (2) sodium monensin Rumenfort (MON1), (3) yeast culture Saccharomyces cerevisiae Biosaf SC47 (LEV), (4) Saccharomyces cerevisiae and probiotic bacteria mix at 1g /animal/day dose (PROB1) and (5) Saccharomyces cerevisiae probiotic bacteria mix at 3g/animal/day dose (PROB2). Treatments did not affect rations DM, OM, NDF and CP digestibilities (P>0,05). Results show that Nellore cattle, castrated or not, feedlot finished receiving rations with high levels of byproducts such as dried citrus pulp and wet corn gluten feed dont have higher performance nor better nutrients digestibility when supplemented with sodium monensin or probiotic microorganisms.

Presented diploma thesis is focused on testing encapsulation methods of enzymes and probiotic bacteria. In the theoretical part a summary of different encapsulation techniques used in food industry is given. Further, materials for encapsulation, above all polysaccharides are presented. Next, some procedures of encapsulation of biopolymers and microorganisms – mainly enzymes and probiotic cultures are discussed. In the experimental part methods for preparation of several types of particles based on polysaccharides and liposomes are introduced. Particles were used for encapsulation of selected hydrolytic enzymes and probiotic strains Bifidobacterium breve a Lactobacillus acidophilus. The encapsulation effectiveness was evaluated by analysis of total proteins and enzyme activities. Particles sizes and their stability in water, in selected model foods and model body fluids were observed, too. According to results obtained in this work it was found that encapsulation of enzymes into polysaccharide particles were succesfull in all types of particles (encapsulation effectivness was more than 50 %). Polysaccharide particles showed a very good stability in body fluids as well as in model foods. As the most suitable materials for enzymes encapsulation chitosan and liposomes were found. Polysaccharide particles were used also for the encapsulation of microorganisms. The stability of particles with lactic acid bacteria was similar to particles containig enzymes, very good stability was verified aslo in model foods and model body fluids. Encapsulation enables long-term stabilization of biologically active compounds as well as posibility of their transport and controlled releasing in gastrointestinal tract. Encapsulation of probiotic bacteria could preserve their viability and long-term survival until the product expiration date. Thus, encapsulation is one of the most promissing procedures for production of foods and food suplements of great quality and high additional value.

Thesis (MScFoodSc)--Stellenbosch University, 2004. / ENGLISH ABSTRACT: The regular consumption of probiotics is becoming a recognized trend in the food industry due to several reported health benefits. A probiotic is defined as a live microbial feed supplement that beneficially affects the host animal by improving its intestinal microbial balance. A wide variety of probiotic food products are available on the South African market and comprise an assortment of fermented milks, as well as lyophilized preparations in tablet or capsule form. Strains of Lactobacillus acidophilus and Bifidobacterium species are mostly used as probiotic microbes in the industry due to their health enhancing effect. The survival of sensitive probiotic microbial species in food matrices are influenced by various factors such as oxygen concentration, pH levels and manufacturing and storage conditions. These should be considered and monitored as the South African food and health regulations stipulate that probiotic microbes should be present at a concentration of 10⁶ cfu.ml ̄ ¹' in order to exert a beneficial effect. Some health benefits are also correlated to specific microbial species and strains and these factors have resulted in the need for the rapid and accurate identification of probiotic microbes present in food products. The probiotic microbes present in probiotic yoghurts and supplements have in the past been identified using traditional methods such as growth on selective media, morphological, physiological and biochemical characteristics. However, even some of the most sophisticated cultural-dependant techniques are not always sufficient for the identification and classification of especially Bifidobacterium, as well as closely related Lactobacillus species. Molecular techniques are more often employed for the rapid and accurate detection, identification and characterization of microbial species present in food products. The aim of this study was to detect and identify the probiotic species present in various commercial South African yoghurts and lyophilized preparations using peR-based DGGE analysis. A 200 bp fragment of the V2-V3 region of the 16S rRNA gene was amplified and the peR fragments were resolved by DGGE. The unique fingerprints obtained for each product were compared to two reference markers A and B in order to identify the bands present. The results obtained were verified by species-specific peR, as well as sequence analyses of bands that could not be identified when compared to the reference markers. Only 54.5% of the South African probiotic yoghurts that were tested did contain all the microbial species as were mentioned on the labels of these products, compared to merely one third (33.3%) of the lyophilized probiotic food supplements. Some Bifidobacterium species were incorrectly identified according to some product labels, while other products contained various microbes that were not mentioned on the label. Sequence analysis confirmed the presence of a potential pathogenic Streptococcus species in one of the yoghurt products and in some instances the probiotic species claimed on the labels were non-scientific and misleading. The data obtained in this study showed that the various South African probiotic products tested were of poor quality and did not conform to the South African regulations. peR-based DGGE analysis proofed to be a valuable approach for the rapid and accurate detection and identification of the microbial species present in South African probiotic products. This could help with future implementation of quality control procedures in order to ensure a reliable and safe probiotic product to the consumer. / AFRIKAANSE OPSOMMING: Die gereelde inname van probiotiese produkte is besig om In erkende tendens in die voedselindustrie te word, as gevolg van verskeie gesondheidsvoordele wat daaraan gekoppel word. In Probiotika word gedefinieer as In voedingsaanvulling wat uit lewendige mikrobes bestaan en wat In voordelige effek op mens of dier het deur In optimale mikrobiese balans in die ingewande te handhaaf. In Wye verskeidenheid probiotiese voedselprodukte is tans beskikbaar op die Suid- Afrikaanse mark. Hierdie bestaan hoofsaaklik uit verskeie gefermenteerde melkprodukte asook 'n reeks tablette en kapsules wat probiotiese mikrobes in gevriesdroogde vorm bevat. Lactobacillus acidophilus tipes en Bifidobacterium spesies word die algemeenste in die voedselindustrie gebruik aangesien hierdie spesifieke mikrobes bekend is om goeie gesondheid te bevorder. Die oorlewing van sensitiewe probiotiese mikrobiese spesies in voedsel matrikse word beïnvloed deur faktore soos suurstof konsentrasie, pH-vlakke en vervaardigings- en opbergings kondisies. Hierdie faktore moet in aanmerking geneem word en verkieslik gemonitor word aangesien die Suid-Afrikaanse voedsel en gesondheids regulasies stipuleer dat probiotiese mikrobes teen In konsentrasie van 10⁶ kolonie vormende eenhede per ml teenwoordig moet wees om In voordelige effek te toon. Sommige gesondheidsvoordele word direk gekoppel aan spesifieke mikrobiese spesies en spesie-tipes. Hierdie faktore het gelei tot In groot aanvraag na vinnige en akkurate metodes vir die identifikasie van probioties mikrobes in voedselprodukte. Die probiotiese mikrobes teenwoordig in probiotiese joghurts en ook die gevriesdroogde vorms in tablette en kapsules, was al geïdentifiseer deur gebruik te maak van tradisionele metodes soos groei op selektiewe media, morfologiese, fisiologiese en biochemiese eienskappe. Selfs van die mees gesofistikeerde kultuur-afhanklike tegnieke is egter nie altyd voldoende vir die identifikasie en klassifikasie van veral Bifidobacterium en na-verwante Lactobacillus spesies nie. Molekulêre metodes word dikwels aangewend vir die vinnige en akkurate deteksie, identifikasie en karakterisering van mikrobes teenwoordig in voedselprodukte. Die doel van hierdie studie was om die probiotiese mikrobes teenwoordig in verskeie Suid-Afrikaanse joghurts en gevriesdroogde aanvullings, te identifiseer deur gebruik te maak van polimerase kettingreaksie (PKR)-gebaseerde denaturerende gradiënt jelelektroforese (DGGE) analise. 'n PKR fragment van 200 bp van die V2-V3 gedeelte van die 16S ribosomale RNS (rRNS) geen is geamplifiseer, en die PKR fragmente is geskei met behulp van DGGE. Die unieke vingerafdrukke wat verkry is vir elke produk is teen twee verwysings merkers A en B vegelyk om die bande teenwoordig in die profiele te identifiseer. Die resultate is bevestig deur spesies-spesifieke PKR en ook deur die ketting volgordes van die DNS fragmente te bepaal wat nie geïdentifiseer kon word deur vergelyking met die verwysings merkers nie. Slegs 54.5% van die Suid-Afrikaanse probiotiese joghurts wat getoets is het al die mikrobiese spesies bevat soos aangedui was op die etikette van hierdie produkte, teenoor slegs 'n derde (33.3%) van die gevriesdroogde voedingsaanvullings. Sekere Bifidobacterium spesies is verkeerd geïdentifiseer op sommige van die produk etikette, terwyl ander produkte verskeie mikrobes bevat het wat nie op die etiket aangedui was nie. 'n Potensiële patogeniese Streptococcus spesie is in een van die joghurt produkte gevind soos bevestig deur DNS kettingvolgorde bepalings. In sommige gevalle was die probiotiese spesienaam wat aangedui is op die etiket onwetenskaplik en misleidend. Die resultate wat uit hierdie studie verkry is dui aan dat die Suid-Afrikaanse probiotiese produkte wat getoets is van 'n swak gehalte is en nie aan die Suid- Afrikaanse regulasies voldoen nie. Daar is getoon dat PKR-gebaseerde DGGE analise 'n waardevolle tegniek kan wees vir die akkurate deteksie en identifisering van die mikrobiese spesies teenwoordig in probiotiese produkte. Dit kan help met die toekomstige implementering van kwaliteitskontrolerings prosedures om 'n mikrobiologiese betroubare en veilige produk aan die verbruiker te verseker.

This Diploma thesis deals with preparation of probiotic cultures Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium breve enriched with prebiotics meant for application in baby food products. Natural extracts from matcha, moringa, young beat, young barley, chlorella and spirulina were selected as prebiotics. The theoretical part is focused on probiotic bacteria, their biological effects and their effects on the child´s body. The experimental part deals with the cultivation of probiotic bacteria with plant extracts, monitoring their viability and stabilization in an encapsulated form. Mixtures of probiotic cells with prebiotics were encapsulated into alginate particles to increase stability. Some of the alginate particles were processed by freeze drying. Mixtures of probiotic cultures with plant extracts were subjected to model human digestion by the action of model digestive juices in unencapsulated, encapsulated and lyophilized form. Selected extracts of plant materials were characterized in terms of amount of total and reducing sugars, total phenolic substances, individual phenolic substances and antioxidant activity. Further, two baby commercial dietary supplements containing probiotics were selected, which were characterized in terms of cell number and viability. Probiotic products were also subjected to model digestion.

Guatemalan medicinal plants were collected and screened for the presence of essential oils using steam distillation. Oil was found in 63 species from 24 families and was tested in tube dilution assays for activity against Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Lactobacillus acidophilus and Candida albicans. Several essential oils were highly active with 20 instances of oils inhibiting the microbes at an MIC of 0.31 µl/ml. Oils were also tested against cancerous and established cell lines using a 15% (v/v) agar-media which was developed to improve essential oil solubility. Assays were performed against three cancer lines: Stomach (AGS: CRL-1739), Skin (A375: CRL-1619), Tongue (CAL27: CRL-2095) and an established Monkey Kidney cell line (Vero C 1008: CRL-1586). Assessment of viability was performed using the Neutral Red assay with results indicating that many of the oils significantly inhibited cancer cell lines in vitro with 24 individual instances producing an IC50 of 0.20 µl/ml or less. Therapeutic indices indicated that many of the highly inhibitory oils were more cytotoxic to cancerous cell lines than to the established cell line.