Esta investigación experimental tuvo como objetivo determinar los cambios histomorfológicos que ocurren en el tejido dentoalveolar de la mandíbula de una progenie de ratas albinas sometidas a dietas con diferentes concentraciones de Riboflavina (B2) e iguales concentraciones del resto de nutrientes, durante la gestación y la lactancia hasta los 28 días de edad. La literatura no reporta estudios que aborden los efectos que se dan en el tejido dentoalveolar por ausencia o deficiencia de B2 desde el punto de vista histológico, y en especial cómo influye en el número de la población celular presente en la unidad dentoalveolar. Se conformaron 4 grupos de ratas albinas (raza Holtzman) con capacidad de reproducción, según el requerimiento de Riboflavina: Grupo A, grupo control, con 100% de B2; Grupo B, con 50% de B2; Grupo C, con 25% de B2 y Grupo D, con 0% de B2. Las crías de las ratas de cada grupo fueron sacrificadas a diferentes edades: 0, 7, 14, 21 y 28 días de nacimiento, y se sometieron luego a un estudio histológico en tinción con H.E. y Masson. Los resultados obtenidos muestran que la deficiencia o la carencia total de B2 ocasiona alteraciones en el crecimiento y desarrollo de las ratas crías, manifestándose una menor talla y disminución del peso corporal. Microscópicamente revelan que la carencia total de Riboflavina (0%) en la dieta materna ocasiona en el tejido dentoalveolar de las crías una población celular menor de ameloblastos, odontoblastos, fibroblastos y cementoblastos, una cantidad muy escasa de fibras colágenas en el ligamento periodontal y un hueso alveolar muy adelgazado, siempre en relación al grupo control A (100% de B2); siendo dichos efectos más notorios a partir de los 7 días en adelante. / The aim of the present experimental study was to determine the histomorphological changes that occur in the dentoalveolar tissue of the jaw of an albino rats’ offspring which were fed with diets containing different concentrations of Riboflavin (B2) and equal concentrations of the rest of nutrients during gestation and nursing until 28 days of birth. Literature does not report studies approaching effects in the dentoalveolar tissue for absence or deficiency of Riboflavin from a histological perspective, especially how it influences on the number of cellular population in the dentoalveolar unit. Four groups of albino rats with reproduction capacity (Holtzman breed) were conformed according to requirements of Riboflavin: Group A, Control Group, with 100% of B2; Group B, with 50% of B2; Group C, with 25% of B2 and Group D, with 0% of B2. The offspring of each group were sacrificed at different ages: 0, 7, 14, 21 and 28 days of birth, and then were submitted to a histological examination applying H.E. and Masson stainings. Results obtained show that deficiency and total lack of B2 cause alterations on growth and development of the offspring, which was expressed as lesser height and decrease in corporal weight. Microscopically they reveal that total lack of Riboflavin (0%) in the mother’s diet causes a smaller population of ameloblasts, odontoblasts, fibroblasts and cementoblasts; a very scarce quantity of collagen fibers in periodontal ligament; and a very thin alveolar bone in the dentoalveolar tissue of the offspring with regard to the Control Group (100% of B2). These effects were more noticeable since 7 days of birth. / Tesis

Esta investigación experimental tuvo como objetivo determinar los cambios histomorfológicos que ocurren en el tejido dentoalveolar de la mandíbula de una progenie de ratas albinas sometidas a dietas con diferentes concentraciones de Riboflavina (B2) e iguales concentraciones del resto de nutrientes, durante la gestación y la lactancia hasta los 28 días de edad. La literatura no reporta estudios que aborden los efectos que se dan en el tejido dentoalveolar por ausencia o deficiencia de B2 desde el punto de vista histológico, y en especial cómo influye en el número de la población celular presente en la unidad dentoalveolar. Se conformaron 4 grupos de ratas albinas (raza Holtzman) con capacidad de reproducción, según el requerimiento de Riboflavina: Grupo A, grupo control, con 100% de B2; Grupo B, con 50% de B2; Grupo C, con 25% de B2 y Grupo D, con 0% de B2. Las crías de las ratas de cada grupo fueron sacrificadas a diferentes edades: 0, 7, 14, 21 y 28 días de nacimiento, y se sometieron luego a un estudio histológico en tinción con H.E. y Masson. Los resultados obtenidos muestran que la deficiencia o la carencia total de B2 ocasiona alteraciones en el crecimiento y desarrollo de las ratas crías, manifestándose una menor talla y disminución del peso corporal. Microscópicamente revelan que la carencia total de Riboflavina (0%) en la dieta materna ocasiona en el tejido dentoalveolar de las crías una población celular menor de ameloblastos, odontoblastos, fibroblastos y cementoblastos, una cantidad muy escasa de fibras colágenas en el ligamento periodontal y un hueso alveolar muy adelgazado, siempre en relación al grupo control A (100% de B2); siendo dichos efectos más notorios a partir de los 7 días en adelante. / The aim of the present experimental study was to determine the histomorphological changes that occur in the dentoalveolar tissue of the jaw of an albino rats’ offspring which were fed with diets containing different concentrations of Riboflavin (B2) and equal concentrations of the rest of nutrients during gestation and nursing until 28 days of birth. Literature does not report studies approaching effects in the dentoalveolar tissue for absence or deficiency of Riboflavin from a histological perspective, especially how it influences on the number of cellular population in the dentoalveolar unit. Four groups of albino rats with reproduction capacity (Holtzman breed) were conformed according to requirements of Riboflavin: Group A, Control Group, with 100% of B2; Group B, with 50% of B2; Group C, with 25% of B2 and Group D, with 0% of B2. The offspring of each group were sacrificed at different ages: 0, 7, 14, 21 and 28 days of birth, and then were submitted to a histological examination applying H.E. and Masson stainings. Results obtained show that deficiency and total lack of B2 cause alterations on growth and development of the offspring, which was expressed as lesser height and decrease in corporal weight. Microscopically they reveal that total lack of Riboflavin (0%) in the mother’s diet causes a smaller population of ameloblasts, odontoblasts, fibroblasts and cementoblasts; a very scarce quantity of collagen fibers in periodontal ligament; and a very thin alveolar bone in the dentoalveolar tissue of the offspring with regard to the Control Group (100% of B2). These effects were more noticeable since 7 days of birth.

As bactérias probióticas, além de diversos possíveis efeitos benéficos à saúde do consumidor, podem apresentar um potencial para bioenriquecer alimentos através da produção de compostos de interesse nutricional e para a saúde durante a etapa de fermentação. Nesse sentido, a produção de vitaminas naturais por bactérias láticas, incluindo as probióticas, emerge como alternativa promissora ao uso de vitaminas sintéticas para a fortificação de alimentos. O uso de substratos vegetais para estimular a produção de vitaminas do grupo B por probióticos compreende uma solução sustentável para o reaproveitamento desse material oriundo, principalmente, do processamento de frutas. Os subprodutos de frutas são compostos por diversas substâncias bioativas e a incorporação desses ingredientes para o desenvolvimento de alimentos com alto valor agregado representa uma solução sustentável para o seu reaproveitamento. Sendo assim, o presente trabalho tem como objetivo avaliar a influência da adição de subproduto de fruta no bioenriquecimento de um produto de soja fermentado tipo frozen yogurt probiótico com riboflavina (vitamina B2). Para este fim, a capacidade de diferentes cepas de Streptococcus thermophilus, Lactobacillus spp. e Bifidobacterium spp. em produzir riboflavina foi avaliada, utilizando-se um meio de cultura desprovido desta vitamina (Riboflavin Assay médium, RAM). Paralelamente, foi avaliada a capacidade dessas cepas em fermentar os subprodutos de uva (Vitis spp.) e de caju (Anacardium occidentale L.). De acordo com os resultados de produção de riboflavina em RAM e de fermentação dos subprodutos de fruta, foram selecionadas as cepas para o desenvolvimento do produto fermentado de soja tipo frozen yogurt (FY). Dentre as cepas testadas, os Streptococcus thermophilus (ST-M6, TA-40 e TH-04) e o probiótico Lactobacillus fermentum PCC apresentaram potencial para produzir riboflavina em meio RAM. As cepas PCC e ST-M6 mostraram-se capazes de fermentar os subprodutos de caju e de uva (atingindo 8,25 e 8,40 log UFC/mL, respectivamente, para PCC e ST-M6 com subproduto de uva e 4,82 e 8,38 log UFC/mL, respectivamente, para PCC e ST-M6 com subproduto de caju em 24h). Por outro lado, as cepas TH-4 e TA-40 fermentaram apenas o subproduto de uva (atingindo 6,87 e 7,24 log UFC/mL, respectivamente, em 24h). Sendo assim, foram selecionadas as cepas de St. Thermophilus ST-M6, TH-4 e TA-40 e a de Lactobacillus fermentum PCC para a aplicação nos produtos visando o seu bioenriquecimento. O FY suplementado com 1% de subproduto de uva foi preparado, utilizando-se as cepas em co-cultura totalizando 3 combinações, ou seja, entre Lb. Fermentum PCC e as 3 cepas de St. Thermophilus. As formulações de FY foram capazes de atingir uma concentração de riboflavina de 8,40 e de 5,18 µg/mL (combinando TA-40 e PCC) em 60 dias, respectivamente, sem e com o subproduto de uva. Já a associação de STM6 e PCC, sem o subproduto, resultou na menor concentração obtida (2,30 µg/mL), atingindo 5,62 µg/mL na presença do subproduto. Dessa maneira aplicação do subproduto de uva para estimular a produção de riboflavina só foi efetiva para a formulação que combinou St. Thermophilus ST-M6 e Lb. Fermentum PCC. / Probiotic bacteria, in addition to several possible beneficial effects on the consumers health, may present a potential to bioenrich food through the production of compounds of nutritional and health interest during fermentation. In this sense, the production of natural vitamins by lactic acid bacteria, including probiotics, emerges as a promising alternative to the use of synthetic vitamins for food fortification. The use of vegetable substrates to stimulate the production of B-group vitamins by probiotics comprises a sustainable solution for the reuse of this material coming mainly from fruit processing. Fruit by-products are composed of various bioactive substances and the incorporation of these ingredients into the development of high value-added foods represents a sustainable solution for their reuse. Therefore, the present study aims to evaluate the influence of the addition of fruit by-product on the bio-enrichment of a probiotic frozen yogurt-like fermented soy product with riboflavin (vitamin B2). For this purpose, the ability of different strains of Streptococcus thermophiles, Lactobacillus spp. and Bifidobacterium spp. in producing riboflavin was evaluated using a culture medium devoid of this vitamin (Riboflavin Assay Medium, RAM). In addition, we evaluated the ability of these strains to ferment the grape (Vitis spp.) and cashew (Anacardium occidentale L.) by-products. According to the results of riboflavin production in RAM and fermentation of fruit by-products, strains were selected for the development of a fermented frozen yogurt-like (FY) soybean product. Among the strains tested, Streptococcus thermophiles (ST-M6, TA-40, and TH-04) and the probiotic Lactobacillus fermentum PCC had the potential to produce riboflavin in RAM medium. The strains PCC and ST-M6 were able to ferment the cashew and the grape byproducts (reaching 8.25 and 8.40 log CFU/mL, respectively, for PCC and ST-M6 with the grape by-product, and 4.82 and 8. 38 log CFU/mL, respectively, for PCC and STM6 with cashew by-product in 24h). On the other hand, TH-4 and TA-40 fermented only the grape by-product (reaching 6.87 and 7.24 log CFU/mL, respectively, in 24h). Therefore, the strains St. thermophiles ST-M6, TH-4, and TA-40 and the strain Lactobacillus fermentum PCC were selected for application in the products for bioenrichment. The FY supplemented with 1% of grape by-product was prepared using the co-culture strains comprising three combinations between Lb. fermentum PCC and the three strains of St. thermophiles. The FY formulations were capable of producing 8.40 and 5.18 µg/mL (combination of TA-40 and PCC) in 60 days, respectively, without and with the grape by-product. However, the association of ST-M6 and CCP, without the by-product, resulted in the lowest concentration obtained (2.30 µg/mL), achieving 5.62 µg/mL in the presence of the by-product. Thus, the application of the grape byproduct in order to stimulate the production of riboflavin was only effective for the formulation that combined St. thermophiles ST-M6 and Lb. fermentum PCC.