La presente investigación se realizó con el fin de evaluar el efecto probiótico de 3 diferentes dosis de Lactobacillus spp. como aditivo sobre parámetros productivos en Cavia porcellus. Se utilizaron 4 grupos de 8 cuyes machos destetados a las 2 semanas cada uno: 3 tratamientos y 1 control. El primer (T1), segundo (T2) y tercer tratamiento (T3) constaron de dosis diarias de 2.5x108, 2.5x109 y 2.5x1010 ufc del probiótico, respectivamente; para el grupo control (TC) suero fisiológico. El probiótico se obtuvo de un yogurt comercial mediante técnicas de aislamiento, identificación y liofilización. La administración del probiótico y el suero fisiológico fue vía oral e individualmente por 7 semanas; además, de recibir concentrado y forraje como alimento. Se empleó un diseño por bloques completos y aleatorizado, las variables en estudio fueron: peso, incremento de peso, conversión alimenticia y rendimiento de carcasa. Los resultados a las 7 semanas fueron: peso final de T1, T2, T3 y TC, 921, 933, 947 y 913 gr; incremento de peso total de T1, T2, T3 y TC, 640.5, 644.75, 632 y 654.75 gr; índice de conversión alimenticia promedio de T1, T2, T3 y TC, 2.90, 2.83, 3.11 y 2.52; y el rendimiento de carcasa promedio de T1, T2, T3 y TC 73.6%, 72.3%, 71.9% y 72.2%, respectivamente. Al análisis estadístico se encontró que no existe diferencia estadísticas significativas entre los grupos tratamiento y control, en las variables estudiadas. A la necropsia no se encontraron lesiones macroscópicas y los grupos tratamiento dieron resultados negativos de cultivos bacterianos para Salmonella spp. en un 100% de los casos, mientras que el grupo control dio un resultado positivo, lo que demuestra que el probiótico posiblemente puede inhibir el crecimiento de Salmonella.The present investigation was realized in order to evaluate the probiotic effect of 3 different doses of Lactobacillus spp. as additive on productive parameters in Cavia porcellus. There were 4 groups of 8 male guinea pigs weaned at 2 weeks each one: 3 treatments and 1 control. The first (T1), the second (T2) and the third (T3) consist of a daily doses of 2.5x108, 2.5x109 y 2.5x1010 cfu of the probiotic respectively; for the control group (TC) only physiological serum. The probiotic was obtained from a commercial yogurt by isolation, identification and freeze-dry techniques. The administration of the probiotic and the serum was individually and orally for 7 weeks, besides receiving concentrate and forage as food. Was used a complete and randomized blocks design, the variables in study were: weight, gain of weight, food conversion and carcass performance. The results at the 7 weeks were: final weight of T1, T2, T3 and TC, 921, 933, 947 and 913 gr; gain of total weight of T1, T2, T3, TC, 640.5, 644.75, 632 and 654.75 gr; average food conversion index of T1, T2, T3 and TC, 2.90, 2.83, 3.11 and 2.52; and average of carcass performance of T1, T2, T3 and TC, 73.6%, 72.3%, 71.9% and 72.2%, respectively. At the statistical analysis were found that there isn’t significant statistic difference between the treatments and control groups in the study variables. At the necropsy there wasn’t macroscopic injuries and the treatments groups show negative results in specific bacterial cultures for Salmonella spp. in 100% of the cases, while the control group have one positive result, which demonstrate that the probiotic can possibly disable Salmonella's growth.

No presente trabalho avaliou-se o efeito antiparasitário de uma preparação probiótica constituída por quatro espécies de lactobacilos, L. casei ATCC 7469, L. plantarum ATCC 8014, L. fermentum ATCC 9338 e L. acidophillus ATCC 4536, no tratamento da eimeriose aviária causada por Eimeria acervulina. Para tanto, foram utilizados 120 animais, da espécie Gallus gallus, machos, com 14 dias de idade, distribuídos em seis grupos, sendo o Grupo 1 constituído por animais tratados apenas com água e ração (controle negativo); Grupo 2 constituído por animais que receberam diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL; Grupo 3 constituído por animais infectados experimentalmente com 2x105 oocistos esporulados de Eimeria acervulina (controle positivo) e Grupo 4 onde os animais receberam diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL durante o período do experimento e após 7 dias do início do tratamento, foram infectados com 2x105 oocistos esporulados de E. acervulina; no Grupo 5 os animais foram infectados com 2x105 oocistos esporulados de E. acervulina e paralelamente receberam uma dose diária da preparação probiótica em estudo contendo 1x108 UFC/mL; no Grupo 6 os animais foram infectados com 2x105 oocistos esporulados de E. acervulina e após o surgimento dos sinais clínicos da infecção passaram a receber diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL, até o final do experimento. Durante 28 dias, semanalmente, amostras de sangue e fezes foram coletadas para determinação de OoPG, testes bioquímicos e parâmetros zootécnicos. Exames de fezes foram realizados diariamente para se determinar os períodos pré-patente e patente do parasita, bem como o efeito da preparação probiótica no combate a infecção. Os resultados demonstraram que a preparação probiótica em estudo reduziu a infecção intestinal causada por Eimeria acervulina, contribuindo para o bem estar animal. Referente aos parâmetros bioquímicos verificou-se efeito significativo sobre o perfil lipídico no 4º dia pós infecção, não sendo observado diferenças significativas para os demais parâmetros. No tocante ao desempenho zootécnico, verificou-se que a preparação probiótica não resultou em diferenças significativas em relação ao ganho de peso, eficiência e conversão alimentar dos animais avaliados. / This study evaluated the antiparasitic effect of a probiotic preparation which consisted of four species of lactobacilli (Lactobacillus casei ATCC 7469, L. plantarum ATCC 8014, L. fermentum ATCC 9338 and L. acidophilus ATCC 4536) in the treatment of eimeriosis in broilers caused by Eimeria acervulina. Therefore, during 28 days, 120 animals of Gallus gallus species, males, 14 days old, were distributed in six groups, as follow: Group 1 the animals received only feed and water \"ad libidum\" (negative control); Group 2 consisted of animals that received a daily dose of probiotic preparation which contained 1x108 CFU/mL; Group 3 the animals were experimentally infected with 2x105 oocysts of Eimeria acervulina (positive control); Group 4 the birds received a daily dose of probiotic preparation containing 1x108 CFU / mL during the experiment and after 7 days of the treatment beginning, these animals were infected with 2x105 oocysts of E. acervulina; Group 5 was composed by animals that were infected with 2x105 oocysts of the parasite and simultaneously received a daily dose of probiotic preparation containing 1x108 CFU / ml, Group 6 the animals were infected with 2x105 oocysts of E. acervulina and after clinical signs of infection were noticed, the birds were treated daily with a dose of the probiotic preparation containing 1x108 CFU / mL until the end of the experiment. During 28 days six blood and feces samples were collected and parameters such as oocyst elimination, histopathological, and biochemical were evaluated. Stool examinations were performed daily to determine the parasite prepatent and patent periods, and the probiotic effect on the infection. The results showed that the probiotic preparation under study reduced the intestinal infection caused by Eimeria acervulina, contributing to animal welfare. Regarding the biochemical parameters there was a significant effect on the lipid profile on the 4th day after infection, and no significant differences were observed for the other parameters. With regard to zootechnical performance, it was found that the probiotic preparation resulted in no significant differences in weight gain, and in feed conversion efficiency.

No presente trabalho avaliou-se o efeito antiparasitário de uma preparação probiótica constituída por quatro espécies de lactobacilos, L. casei ATCC 7469, L. plantarum ATCC 8014, L. fermentum ATCC 9338 e L. acidophillus ATCC 4536, no tratamento da eimeriose aviária causada por Eimeria acervulina. Para tanto, foram utilizados 120 animais, da espécie Gallus gallus, machos, com 14 dias de idade, distribuídos em seis grupos, sendo o Grupo 1 constituído por animais tratados apenas com água e ração (controle negativo); Grupo 2 constituído por animais que receberam diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL; Grupo 3 constituído por animais infectados experimentalmente com 2x105 oocistos esporulados de Eimeria acervulina (controle positivo) e Grupo 4 onde os animais receberam diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL durante o período do experimento e após 7 dias do início do tratamento, foram infectados com 2x105 oocistos esporulados de E. acervulina; no Grupo 5 os animais foram infectados com 2x105 oocistos esporulados de E. acervulina e paralelamente receberam uma dose diária da preparação probiótica em estudo contendo 1x108 UFC/mL; no Grupo 6 os animais foram infectados com 2x105 oocistos esporulados de E. acervulina e após o surgimento dos sinais clínicos da infecção passaram a receber diariamente uma dose da preparação probiótica contendo 1x108 UFC/mL, até o final do experimento. Durante 28 dias, semanalmente, amostras de sangue e fezes foram coletadas para determinação de OoPG, testes bioquímicos e parâmetros zootécnicos. Exames de fezes foram realizados diariamente para se determinar os períodos pré-patente e patente do parasita, bem como o efeito da preparação probiótica no combate a infecção. Os resultados demonstraram que a preparação probiótica em estudo reduziu a infecção intestinal causada por Eimeria acervulina, contribuindo para o bem estar animal. Referente aos parâmetros bioquímicos verificou-se efeito significativo sobre o perfil lipídico no 4º dia pós infecção, não sendo observado diferenças significativas para os demais parâmetros. No tocante ao desempenho zootécnico, verificou-se que a preparação probiótica não resultou em diferenças significativas em relação ao ganho de peso, eficiência e conversão alimentar dos animais avaliados. / This study evaluated the antiparasitic effect of a probiotic preparation which consisted of four species of lactobacilli (Lactobacillus casei ATCC 7469, L. plantarum ATCC 8014, L. fermentum ATCC 9338 and L. acidophilus ATCC 4536) in the treatment of eimeriosis in broilers caused by Eimeria acervulina. Therefore, during 28 days, 120 animals of Gallus gallus species, males, 14 days old, were distributed in six groups, as follow: Group 1 the animals received only feed and water \"ad libidum\" (negative control); Group 2 consisted of animals that received a daily dose of probiotic preparation which contained 1x108 CFU/mL; Group 3 the animals were experimentally infected with 2x105 oocysts of Eimeria acervulina (positive control); Group 4 the birds received a daily dose of probiotic preparation containing 1x108 CFU / mL during the experiment and after 7 days of the treatment beginning, these animals were infected with 2x105 oocysts of E. acervulina; Group 5 was composed by animals that were infected with 2x105 oocysts of the parasite and simultaneously received a daily dose of probiotic preparation containing 1x108 CFU / ml, Group 6 the animals were infected with 2x105 oocysts of E. acervulina and after clinical signs of infection were noticed, the birds were treated daily with a dose of the probiotic preparation containing 1x108 CFU / mL until the end of the experiment. During 28 days six blood and feces samples were collected and parameters such as oocyst elimination, histopathological, and biochemical were evaluated. Stool examinations were performed daily to determine the parasite prepatent and patent periods, and the probiotic effect on the infection. The results showed that the probiotic preparation under study reduced the intestinal infection caused by Eimeria acervulina, contributing to animal welfare. Regarding the biochemical parameters there was a significant effect on the lipid profile on the 4th day after infection, and no significant differences were observed for the other parameters. With regard to zootechnical performance, it was found that the probiotic preparation resulted in no significant differences in weight gain, and in feed conversion efficiency.

De nos jours, l’obésité est un problème majeur de santé publique dont la prévalence mondiale ne cesse d’augmenter. Ainsi selon la National Health and Nutrition Examination Surveys (NHANES), le pourcentage d’adultes obèses aux Etats-Unis a presque doublé entre 1976-1980 et 1999-2002. De plus, pour la première fois dans l’histoire des Etats-Unis, la longévité de l’actuelle génération d’enfants est estimée à la baisse. Cette augmentation de l’obésité résulte à la fois de prédispositions génétiques et de facteurs environnementaux tels que l’accès à la nourriture, le tissu social, le régime alimentaire ou encore le manque d’activité physique. , Aussi, de récentes études ont suggéré que la flore microbienne intestinale jouait un rôle majeur dans la conversion énergétique des nutriments et son stockage. Bien que très peu d’expériences aient été réalisées, le rôle des Lactobacillus spp. a déjà été évoqué dans la prise de poids chez l’homme. Deux importantes études, l’une menée au Danemark et l’autre au sein même de notre laboratoire, ont montré que les Lactobacillus spp. étaient présents en quantité plus importante chez les sujets obèses par rapport aux contrôles et que leur taux corrélaient à la glycémie chez des patients diabétiques de type II (obèses). L’effet des Lactobacillus spp sur la prise de poids a été beaucoup plus étudié chez les animaux. En effet, l’agriculture représente l’un des domaines majeur d’utilisation des antibiotiques comme « growth promoters », notamment l’avoparcin qui empêche le développement de nombreuses bactéries dont des Gram positives comme les Actinobacteria et les Firmicutes à l’exception toutefois des Lactobacillus spp. Les probiotiques sont aussi utilisés comme « growth promoters » et il a été montré que l’addition de 106 Lactobacillus spp. dans les régimes alimentaires d’animaux permettait de modifier leur flore microbienne intestinale et d’entrainer un gain de poids important. A partir de toutes ces constatations, nous avons émis l’hypothèse que les bactéries pouvaient jouer un rôle dans l’obésité via une modification de la composition de la flore intestinale. Il nous est également apparu essentiel d’évaluer l’effet de l’addition de concentration bactérienne importante dans l’alimentation quotidienne. L’objectif de notre étude est ainsi d’évaluer le rôle des Lactobacillus spp. en tant que facteur de croissance et d’évaluer leur impact sur la prise de poids. / The prevalence of obesity is a major world health problem that is rapidly increasing. In the USA, according to the National Health and Nutrition Examination Surveys (NHANES), the percentage of obese adults nearly doubled between NHANES 1976–1980 and NHANES 1999– 2002. Moreover, because of the increase in prevalence of American childhood obesity, the current children generation is predicted to be the first in United States to see a decrease in longevity. Obesity results from a mixture of genetic background and environmental factors including food availability, social networks, diet, and physical activity. Recent evidence suggests that gut microbiota plays a major role in energy intake, conversion and storage. Although very few reports have investigated gut microbiota and its association with obesity, several evidences support a role of Lactobacillus spp. in weight gain within human. Lactobacillus spp. were found at higher levels among obese subjects in two major studies, one conducted by us and one made in Denmark. Moreover, the rate of Lactobacillus spp. was correlated with blood glucose in subjects with type II diabetes (obese). In agriculture avoparcin which is one of the most used antibiotics to cause weight gain in animals, is effective on most bacteria including Gram-positive Actinobacteria and Firmicutes, with the notable exception on some Lactobacillus spp.. In addition, probiotics are also used as growth promoters and the addition of even 106 Lactobacillus spp in the diet of farm animals results in gut microbiota changes and weight gain. As a result, we hypothesized that bacteria could play a role in the obesity pandemic notably with the ingestion of probiotics that modified the gut microbiota structure. In addition, we stressed the necessity for further investigations to assess the effects of routinely adding high amounts of bacteria in food. The objective of our study was to evaluate the putative role of Lactobacillus sp. in growth promotion.

Submitted by Nathália Faria da Silva (nathaliafsilva.ufv@gmail.com) on 2017-07-07T17:48:43Z No. of bitstreams: 1 texto completo.pdf: 587612 bytes, checksum: 559af73c6466c747ea1f836f3b491147 (MD5) / Made available in DSpace on 2017-07-07T17:48:43Z (GMT). No. of bitstreams: 1 texto completo.pdf: 587612 bytes, checksum: 559af73c6466c747ea1f836f3b491147 (MD5) Previous issue date: 2001-03-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Fundação de Amparo à Pesquisa do Estado de Minas Gerais / Probióticos, na forma de produtos lácteos fermentados ou não, são conhecidos como alimentos funcionais e têm sido recomendados como adjunto dietético para indivíduos com hipercolesterolemia. Com o objetivo de avaliar seu efeito na modulação dos níveis de colesterol sérico e na taxa de translocação de Lactobacillus spp. para o baço, fígado, coração e rins, foi conduzido uma experimentação com duração de 56 dias, utilizando-se 170 ratos machos Wistar, com peso médio inicial de 250±32g, distribuídos em quatro tratamentos e alojados individualmente. Os grupos experimentais foram: Padrão; Controle; LDR e P. O grupo Padrão recebeu a dieta AIN-93G durante todo o período experimental. Os demais grupos receberam por 56 dias a mesma dieta acrescida de 1% de colesterol e 0,1% de ácido cólico. Do 15° ao 28° dia (após um período inicial de 14 dias de indução de hipercolesterolemia) o grupo LDR recebeu suplementação de 0,1 mL/dia/animal de leite desnatado reconstituído a 10% de sólidos não gordurosos e o grupo P recebeu 0,1 mL/dia/animal de probiótico na 10 forma de um concentrado de células contendo 10 UFC/mL de Lactobacillus acidophilus NCFM. Avaliou-se o colesterol total, LDL- colesterol e HDL-colesterol nos 28°, 42° e 56° dias de experimento (0, 14 e 28 dias após o término da administração do probiótico, respectivamente). Determinou-se também a taxa de translocação de Lactobacillus spp., nos mesmos tempos, no baço, fígado, coração e rins dos animais. O ganho de peso e a ingestão alimentar dos animais foram monitorados semanalmente, calculando -se assim, o coeficiente de eficiência alimentar (CEA). Não foi observada diferença significativa (P>0,05) entre os grupos, quanto ao consumo alimentar, ganho de peso e CEA aos 56 dias. O acréscimo dietético de 1% de colesterol cristalino e 0,1% de ácido cólico não promoveu hipercolesterolemia nos animais, evidenciado pela diferença não significativa (P>0,05) entre os grupos Padrão e Controle em relação aos níveis de colesterol total, LDL- colesterol e HDL-colesterol após 14 dias de dieta hipercolesterolêmica. No entanto, provocou aumento significativo do peso e alteração da coloração normal do fígado (de vermelho intenso para vermelho amarelado) devido ao acúmulo de gordura neste órgão, nos animais que receberam dieta rica em colesterol comparado ao grupo Padrão. O consumo de L. acidophilus pelos animais, nas concentrações indicadas, não alterou seus níveis séricos de colesterol e frações (P>0,05), pois estes se mantiveram normocolesterolêmicos. Constatou-se translocação de Lactobacillus spp. para o baço, fígado, coração e rins dos animais que receberam suplementação com L. acidophilus (grupo P). Nos órgãos dos animais dos grupos Padrão, Controle e LDR não foi observado crescimento nos tempos avaliados. Constatou-se diferença significativa (P<0,05) na contagem de células microbianas translocadas para o baço em comparação aos demais órgãos analisados. Entre o fígado e o coração não foi observada diferença significativa (P>0,05), mas ambos diferiram estatisticamente dos rins (P<0,05). As contagens, 2 2 em UFC/órgão, foram; 8,6 x 10 ; 4,8 x 10 ; 4,7 x 10 e 1,3 x 10 para o baço, fígado, coração e rins, respectivamente. Observou-se eliminação constante das células translocadas, após o término da administração do probiótico. Vinte e oito dias após o consumo do probiótico, de acordo com o modelo de regressão ajustado, a contagem de células viáveis indicou maior eliminação dos microrganismos translocados do baço (69,8%), seguido do coração (61,7%), fígado (57,0%) e rins (37,8%). O tempo para completa eliminação dos microrganismos translocados, de acordo com o modelo de regressão ajustado, permanecendo a tendência verificada no intervalo de tempo estudado (28 dias), corresponderia a 40, 45, 49 e 74 dias, para o baço, coração, fígado e rins, respectivamente. Sugere-se que, ao se desenvolver um probiótico, além da estirpe do microrganismo, do número de células e da freqüência de dosagem, devem ser definidas as taxas de translocação e de eliminação (“clearing”) destes microrganismos em modelos animais, uma vez que se desconhece como este processo poderá afetar o hospedeiro. / Probiotics, as milk fermented products or not, are known as functional foods and have been recommended as dietary adjunct for individuals with high serum cholesterol levels. To evaluate probiotics effects on the modulation of serum cholesterol and on translocation rate of Lactobacillus spp. for the spleen, liver, heart and kidneys, an experiment was carried out during 56 days, using 170 Wistar male rats, distributed in four treatments and individually housed, with initial average weight of 250±32g. The experimental groups were: Standard; Control; LDR and P. The Standard group received the AIN- 93G diet during the whole experimental period. The other groups received for 56 days the AIN -93G diet added of 1% of cholesterol and th th 0,1% of cholic acid. Of the 15th to the 28th day (after an initial period of 14 days of hipercolesterolemia induction) the LDR group received the supplement of 0,1 mL/day/animal of reconstituted skimmed milk at 10% of non fat solids and, the group P received 0,1 mL/day/animal of a 10 probiotic in the form of a concentrate of cells contends 10 UFC/mL of Lactobacillus acidophilus NCFM. Total cholesterol, high density lipoprotein (HDL) and low density lipoprotein (LDL) were evaluated th th th at 28th, 42th and 56th days of experiment (0, 14 and 28 days after the end of probiotic administration respectively). Translocation rate of Lactobacillus spp. was monitored at the same times, in spleen, liver, heart and kidney of all animals. The weight gain and alimentary ingestion were weekly verified, in order to calculate the coefficient of alimentary conversion (CEA). No significant differences (P>0,05) ware observed between groups, relative to alimentary consume, weight th increase and CEA at the 56th day. The dietary increment of 1% of crystalline cholesterol and 0,1% of cholic acid did not promote hipercholesterolemia in the animals, evidenced by the non significant difference (P>0,05), among the groups Standart and Control, of levels of total cholesterol, LDL-cholesterol and HDL-cholesterol levels after 14 days receiving hipercholesterolemic diet. However it provoked significant increase of the weight and alteration of normal coloration of liver (of red intense for red yellowish) due to fat accumulation in this organ, in the animals that received rich diet in cholesterol compared to the Standart group. Consumption of L. acidophilus, in the suitable concentrations, did not altered serum cholesterol levels and its fractions (P>0,05) of the animals, once they stayed normocholesterolemic. Translocation of Lactobacillus spp was verified at the spleen, liver, heart and kidneys of animals supplemented with L. acidophilus (group P). The evaluation of the organs from the Standard, Control and LDR groups did not indicate the presence of Lactobacillus. Significant difference was verified (P<0,05) in the translocated microbial cells count for the spleen in comparison to the other analyzed organs. Significant difference (P>0,05) was not observed between the liver and the heart translocated microbial cells count, but both differed statisticaly of the kidneys (P<0,05). The average translocation, expressed as CFU/organ, were 8,6 x 10 ; 4,8 x 10 2 ; 4,7 x 10 2 and 1,3 x 10 2 for spleen, liver, heart and kidney, respectively. A constant elimination of the translocated cells was observed after the end of the probiotic administration. The eliminated translocated cell numbers were constantly during the evaluation period. Twenty-eight days after probiotics consumption, according to the adjusted regression model, the viable cells count indicated larger elimination of translocated microrganismis from spleen (69,8%), heart (61,7%), liver (57,0%) and kidneys (37,8%). The time required for the complete elimination of translocated Lactobacillus spp., according to the adjusted regression model and supposing that the elimination of translocated cells by the organism stays constant, correspond to 40; 45; 49 and 74 days, respectively for spleen, heart, liver and kidneys. Translocation and elimination rate (“clearing”) of microrganisms in animal models are suggested as important parameters on the development of probiotic products. The dose frequency, number of viable cells and microorganism lineage must be analyzed and selection of strains that do not present translocation must be considered, since the implication of this process in the host is not known.

Submitted by FELIPE EDUARDO DE OLIVEIRA (felipe.eoliveira@ymail.com) on 2017-04-07T14:51:33Z No. of bitstreams: 1 TESE FELIPE CORRIGIDA.pdf: 5002341 bytes, checksum: 809ad9cdf3b52f15db51efb7247aa675 (MD5) / Approved for entry into archive by Luiz Galeffi (luizgaleffi@gmail.com) on 2017-04-17T14:10:43Z (GMT) No. of bitstreams: 1 oliveira_fe_dr_sjc.pdf: 5002341 bytes, checksum: 809ad9cdf3b52f15db51efb7247aa675 (MD5) / Made available in DSpace on 2017-04-17T14:10:43Z (GMT). No. of bitstreams: 1 oliveira_fe_dr_sjc.pdf: 5002341 bytes, checksum: 809ad9cdf3b52f15db51efb7247aa675 (MD5) Previous issue date: 2017-02-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Muitos produtos probióticos são compostos pela associação de lactobacilos no intuito de potencializar seu efeito benéfico. Assim, o objetivo deste estudo foi avaliar a imunomodulação e atividade anti-Candida causada por diferentes combinações de suspensões probióticas em macrófagos e em Galleria mellonella. Lactobacillus rhamnosus (LR), L. acidophilus (LA), e L. paracasei (LP) foram suspendidos para obtenção das suspensões LR, LA, LP, LR+LA, LR+LP, LA+LP e LR+LA+LP. Foram utilizadas suspensões de Candida albicans (CA) ATCC 18804, C. krusei (CK) ATCC 6258 e C. glabrata (CG) ATCC 9030. Macrófagos de camundongo (RAW 264.7) foram ativados por cada suspensão de lactobacilos e desafiados por cada uma das cepas fúngicas. Foram investigadas taxa de fagocitose, produção de citocinas e óxido nítrico e a viabilidade celular. A melhor suspensão probiótica para cada cepa fúngica seria usada nos testes in vivo, com G. mellonella, no entanto, devido à baixa virulência das espécies não-albicans, os testes in vivo prosseguiram com CA e LR. Duas cepas clínicas de CA foram acrescentadas: 21 e 60. Suspensões de LR foram injetadas na hemolinfa das larvas 24 h antes do desafio com as cepas fúngicas. Foram determinadas curva de sobrevivência, contagem de UFC/mL das cepas de C. albicans e de hemócitos. Os resultados foram analisados estatisticamente de acordo com a distribuição normal (ANOVA e Tukey ou Kruskal-Wallis e Dunn) e a curva de sobrevivência pelo teste de Log-rank (Mantel-Cox), p<0,05. As melhores suspensões para CA, CK e CG foram LR (com melhores resultados no perfil de citocinas), LP e LA (com melhores resultados sobre a fagocitose), respectivamente. Em G. mellonella, a suspensão de LR proporcionou aumento da percentagem de sobrevivência e do número de hemócitos e diminuição da contagem de UFC/mL em todos os grupos com diferenças estatisticamente significantes. Sendo assim, pode-se concluir que as suspensões de lactobacilos possuem potencial imunomodulador cepa dependente mas sua mistura não favorece essa qualidade. Além disso, possuem atividade anti-Candida, demonstrada pela diminuição na contagem de Candida. / Many probiotic products are composed by association of lactobacilli in order to potentiate their beneficial effect. Therefore, the objective of this study was evaluating the immunomodulation and anti-Candida activity caused by different combinations of probiotic suspensions in macrophages and in Galleria mellonella. Lactobacillus rhamnosus (LR), L. acidophilus (LA) and L. paracasei (LP) were suspended to obtain the suspensions LR, LA, LP, LR + LA, LR + LP, LA + LP and LR + LA + LP. Suspensions of Candida albicans (CA) ATCC 18804, C. krusei (CK) ATCC 6258 and C. glabrata (CG) ATCC 9030 were used. Mouse macrophages (RAW 264.7) were activated by each suspension of lactobacilli and then challenged by each fungal strain. Phagocytosis, cytokine and nitric oxide production and cell viability were investigated. The best lacobacilli suspension for each fungal strain would be used in the in vivo assays with G. mellonella, however, because of low virulence of the non-albicans species, in vivo tests were performed with CA and LR. Two clinical strains of CA were added: 21 and 60. LR suspensions were injected into the haemolymph of the larvae 24 h before challenging with fungal strains. Survival curve, CA strains CFU/mL counting and hemocytes were determined. The results were statistically analyzed according to their normal distribution (ANOVA and Tukey or Kruskal-Wallis and Dunn) and the survival curve by Log-rank test (Mantel-Cox), p <0.05. The best suspensions for CA, CK and CG were LR (which obtained the best results in the cytokine profile), LP and LA (which obtained the best results on phagocytosis), respectively. In G. mellonella, LR suspension increased percentage survival and hemocyte counting and decreased CFU/mL counting in all groups, with statistically significant differences in the results. Thus, it can be concluded that the suspensions of lactobacilli have immunomodulatory potential strain-dependent but their association does not favor this quality. Besides, they showed anti-Candida activity, demonstrated by the decrease of Candida counting.

Darbo tikslas: ištirti apsauginių kultūrų panaudojimo mėsos pusgaminių gamyboje galimybes, gerinant saugą ir kokybę. Darbo uždaviniai: įsisavinti apsauginių kultūrų panaudojimo mėsos pusgaminių gamyboje būdus, pusgaminių kokybinių ir saugos rodiklių nustatymo metodus; ištirti apsauginių bakterijų kultūrų: Lactobacillus spp. Staphylococcus xylosus, poveikį mėsos pusgaminių mikrobiologiniams ir fizikiniams cheminiams rodikliams, įvertinti jų saugą ir kokybę; ištirti termiškai apdorotų mėsos pusgaminių su apsauginėmis bakterinėmis kultūromis Lactobacillus spp., Staphylococcus xylosus juslines savybes; palyginti įprastų ir gautų mėsos pusgaminių su apsauginėmis kultūromis kokybę ir saugą, juslines savybes, vartojimo terminą; apdoroti statistiškai gautus duomenis, apibendrinti rezultatus ir pateikti išvadas. Rezultatai: priedas su apsauginėmis kultūromis Lactobacillus spp., Staphylococcus xylosus visiškai išnaikino E. Coli jautienos farše, supakuotame į vakuuminę pakuotę (p ≤ 0,01), modifikuotų dujų atmosferą (p ≤ 0,01), laikomo aerobinėmis sąlygomis (p ≤ 0,001) lyginant su kontrole. Apsauginių kultūros ženkliai sumažino bendrą aerobinių mikroorganizmų skaičių lyginant su kontrole (p ≤ 0,001). Apsauginių kultūrų įterpimas sumažino biogeninių aminų tiramino, spermidino ir spermino kiekius iki mažesnių už aptikimo ribą kiekių (< 5 mg/kg), tačiau mėginiuose su apsauginių kultūrų priedu, laikomuose aerobinėmis sąlygomis, nustatytas padidėjęs fenilalanino kiekis, lyginant su kontrole... [toliau žr. visą tekstą] / The aim of the work was to investigate the possibilities of using protective cultures in the manufacture of semi-finished meat in improving safety and quality. The tasks of the work were to master ways of using protective cultures in production of semi-finished meat; methods for determining quality and safety indicators of semi – finished meat; to investigate the impact of protective bacterial cultures: Lactobacillus spp., Staphylococcus xylosus to microbiological and physico-chemical parameters of semi – finished meat in order to ensure their safety and quality; to investigate organoleptic properties of the heat-treated semi – finished meat with a protective bacterial cultures Lactobacillus spp., Staphylococcus xylosus; to compare quality and safety, organoleptic characteristics, expiry date of conventional and meat preparations with protective cultures; to process statistically obtained data, summarize the results and present findings. Results: • Additive with protective cultures Lactobacillus spp., Staphylococcus xylosus completely destroyed E. coli in vacuum packed minced beef (p ≤ 0,01), in modified gas atmosphere (p ≤ 0,01), stored under aerobic conditions (p ≤ 0,001) compare with the controls. • Protective cultures significantly reduced the total number of aerobic microorganisms compare with controls (p ≤ 0,001). • Insertion of protective cultures reduced biogenic amines tyramine, spermidine and spermine below the detection limit (<5 mg / kg), however, samples... [to full text]

Submitted by Viviane Lima da Cunha (viviane@biblioteca.ufpb.br) on 2016-02-22T14:57:38Z No. of bitstreams: 1 arquivototal.pdf: 3702710 bytes, checksum: 9dee0e28c3be22319d303a2bc4134ddf (MD5) / Made available in DSpace on 2016-02-22T14:57:38Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3702710 bytes, checksum: 9dee0e28c3be22319d303a2bc4134ddf (MD5) Previous issue date: 2015-07-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Products such as cheeses, yoghurts and milk drinks can be obtained from goat's milk are an alternative to the increase in consumption and value of goatderived products. This study aimed to produce ricotta cheese from goat whey and evaluate the effects of incorporating probiotic strains Bifidobacterium lactis BB-12 and Lactobacillus acidophilus LA-5 on technological parameters, physicochemical and sensory cheese during storage refrigerated for seven days, and the protective effect of goat ricotta on the survival of probiotics tested for exposure to simulated gastrointestinal conditions. A Ricotta goat was prepared based on goat cheese whey (main ingredient) with addition of whole goat milk (20%). The incorporation of probiotics did not influence the income and syneresis of goat ricotta. During the rated cold storage period, the strains L. acidophilus LA-5, and B. lactis BB-12 had scores of about 6 log CFU / g. Samples ricotta containing the probiotic strains showed the lowest values for lactose and the highest values for lactic acid, besides having greater hardness and lower brightness during storage when compared to samples without probiotics. There were no differences in the fatty acid profile of goat ricotta with or without probiotics. All samples of ricotta were sensorially described as "soft cheese" with "homogeneous texture", however, the samples added L. acidophilus LA-5 or B. lactis BB-12 were described as "more acidic". At the end of exposure to simulated gastrointestinal conditions, the tested probiotic strains showed scores of approximately 6 log CFU / g, when incorporated in goat ricotta. These results show the possibility of using L. acidophilus LA-5, and B. lactis BB-12 in the preparation of goat ricotta, considering that such strains showed no negative impact on the general quality characteristics of the product. Further, goats show the ricotta as a satisfactory matrix for the continued viability of probiotics tested during storage as well as when exposed to the harsh conditions found throughout the human gastrointestinal tract. / Produtos como queijos, iogurtes e bebidas lácteas podem ser obtidos a partir do leite de cabra sendo uma alternativa para o aumento no consumo e agregação de valor de produtos de origem caprina. O presente estudo teve como objetivo produzir queijos ricota a partir de soro de leite caprino e avaliar os efeitos da incorporação de cepas probióticas Bifidobacterium lactis BB-12 e Lactobacillus acidophilus LA-5 sobre os parâmetros tecnológicos, físicoquímicos e sensoriais do queijo durante armazenamento refrigerado de sete dias, bem como o efeito protetor da ricota caprina sobre a sobrevivência dos probióticos testados durante exposição a condições gastrointestinais simuladas. A ricota caprina foi elaborada tendo como base soro de queijo caprino (ingrediente principal) com adição de leite integral de cabra (20%). A incorporação dos probióticos não influenciou no rendimento e sinérese da ricota caprina. Durante o período de armazenamento refrigerado avaliado, as cepas de L. acidophilus LA-5 e B. lactis BB-12 apresentaram contagens em torno de 6 log UFC/g. As amostras de ricota contendo as cepas probióticas apresentaram os menores valores para lactose e os maiores valores para ácido láctico, além de apresentarem maior dureza e menor brilho durante a armazenagem, quando comparados com amostras sem probióticos. Não foram observadas diferenças no perfil de ácidos graxos da ricota caprina com ou sem probióticos. Todas as amostras de ricota foram sensorialmente descritas como sendo “queijo macio” com “textura homogênea”, porém, as amostras adicionadas de L. acidophilus LA-5 ou B. lactis BB-12 foram descritas como sendo “mais ácidas”. Ao final da exposição às condições gastrointestinais simuladas, as cepas probióticas testadas apresentaram contagens de aproximadamente 6 log UFC/g, quando incorporadas na ricota caprina. Estes resultados mostram a possibilidade do uso de L. acidophilus LA-5 e B. lactis BB-12 na elaboração de ricota caprina, tendo em vista que tais cepas não apresentaram impacto negativo sobre as características gerais de qualidade do produto. Ainda, evidenciam a ricota caprina como uma matriz satisfatória para a manutenção da viabilidade dos probióticos ensaiados durante o armazenamento, bem como quando expostos às condições adversas encontradas ao longo do trato gastrointestinal humano.

Amino acids derived from the degradation of casein in cheese serve as precursors for the generation of desirable and undesirable flavor compounds. Microbial degradation of aromatic amino acids is associated with the formation of aroma compounds that impart putrid-fecal, barny-utensil, and floral off-flavors in cheese, but pathways for their production had not been established. This study investigated Tyr and Phe catabolism by Lactobacillus casei and Lactobacillus helveticus cheese flavor adjuncts under simulated Cheddar cheese-ripening conditions (pH 5.2, 4% NaCl, 15°C, no sugar). Enzyme assays of cell-free extracts and micellar electrokinetic capillary chromatography of supernatants indicated that L. casei and L. helveticus strains catabolize Tyr and Phe by successive transamination and dehydrogenation reactions. Major products of Tyr and Phe catabolism included off-flavor compounds formed by chemical degradation of the α-keto acids, produced by transamination, and aromatic α-hydroxy acids derived from α-keto acids by α-hydroxy acid dehydrogenases. Action of Lacrococcus lactis aminotransferase enzymes on Trp, Tyr, and Phe also leads to the formation of α-keto acids, but unlike lactobacilli, the former bacteria do not express dehydrogenase activity under cheese-like conditions (pH 5.2, 4% NaCl, 15°C, no sugar). Since aromatic α-keto acids may degrade spontaneously into undesirable flavor compounds, α-hydroxy acid dehydrogenases may be useful in controlling off-flavor development via diversion of chemically labile α-keto acids to more stable a-hydroxy acids. To test this hypothesis, we investigated the effect of D-hydroxyisocaproate dehydrogenase overexpression by a L. casei adjunct on chemical and sensory properties of reduced-fat Cheddar cheese made with and without addition of 20 mM α-ketoglutarate. The D-hydroxyisocaproic acid dehydrogenase gene (D-HicDH) was cloned into a high copy number vector pTRKH2 and transformed into L. casei ATCC334. Reduced-fat Cheddar cheeses were made with Lactococcus lactis starter only, starter + L. casei ATCC334 with pTRKH2, and starter + L. casei ATCC334 with pTRKH2: D-HicDH, and then volatile analysis was performed by gas chromatography and mass spectrometry. Statistical analysis of volatile data after 3 mo of ripening at 7°C showed profiles of ketones, aldehydes, alcohols, esters, sulfur compounds, and benzaldehyde were significantly altered by culture treatments and α-ketoglutarate addition, and these treatments also affected sensory flavor attributes of experimental cheeses. Results also indicated overexpression of D-hydroxyisocaproic acid dehydrogenase can divert labile α-keto acids into more stable compounds, but the overall effect seemed to diminish both beneficial and detrimental flavor notes.

For production of double moulded chesses are used lactic acid bacteria, which can be present in a form of non-starter lactic acid bacteria or as starter or adjunct culture. Genera Lactobacillus spp. and Enterococcus spp. are prevalent microorganisms present in these cultures. Of course, these microorganisms are for us interesting because of their possibility of coagulation, proteolytic possibility, probiotic function and antibiotic resistance, but especially because of their decarboxylation abilities. Bacteria contain decarboxylation enzymes, which are able to decarboxylized free amino acid, which rising at proteolysis during process of manufacturing and cheese ripening. Biogenic amines are the result of proteolytic activity. Biogenic amines occur practically in all foodstuffs as a common product of metabolic processes. BA are mainly presented in fermented food (cheeses), where rice just microbial action. Typical representatives of biogenic amines, which occurs in double moulded cheeses (Sedlčanský Vltavín, Bresse bleu) and in blue cheeses (Bleu des Causses, Bleu d'Auvergne) are cadaverine, putrescine, tyramine a 2 fenylethylamine and in much smaller amount histamine, spermidine and spermine too. On assessment concentration of BA is used high pressure liquid chromatography with reverse phase (RP HPLC) with utilizing simple direct derivatization with dansyl chloride and detection by UV VIS detector.