Orientador: Ana Lia Anbinder / Banca: Maria Aparecida Neves Jardini / Banca: Lucilene Fernandes Ricardo / Resumo: A doença periodontal (DP) é caracterizada pela destruição dos tecidos de proteção e suporte dentário, osso alveolar, ligamento periodontal, cemento e gengiva, e é a principal causa de perda dentária na população adulta, um grande problema socioeconômico e de saúde pública. Com o aumento da resistência antibiótica, existe interesse no uso de outras substâncias para o combate de doenças infecciosas, como os probióticos. O uso de probióticos na prevenção e tratamento de doença periodontal em animais tem apresentado bons resultados. O estudo dos efeitos do Lactobacillus reuteri viável e inativado (paraprobiótico) pode esclarecer mecanismos de ação e possibilitar o uso comercial de outras preparações, dado que os paraprobióticos teriam produção facilitada. O objetivo deste trabalho foi analisar os efeitos de L. reuteri, viável e inativado, em doença periodontal induzida em ratos. Foram utilizados 32 ratos machos, divididos em 4 grupos: (1) Grupo Controle: com saúde periodontal (n=8); (2) Grupo LIP: periodontite induzida (n=8); (3) Grupo LIP/PRO: periodontite induzida e probiótico (n=8); e (4) Grupo LIP/PARAPRO: periodontite induzida e paraprobiótico (n=8). A indução da doença periodontal foi realizada com a inserção de fio de algodão ao redor dos primeiros molares inferiores e foi realizado o tratamento com L. reuteri viável ou inativado pelo calor, via oral, diariamente, 30 dias antes da ligadura e 14 dias após, quando ocorreu a eutanásia. As hemimandíbulas direitas foram removid... (Resumo completo, clicar acesso eletrônico abaixo) / Periodontal disease (PD) is characterized by the destruction of dental protection and support tissues, alveolar bone, periodontal ligament, cementum and gingiva, and is the major cause of tooth loss in the adult population, a major socioeconomic and public health problem. With the increase of antibiotic resistance, there is interest in the use of other substances to combat infectious diseases, such as probiotics. The use of probiotics in the prevention and treatment of periodontal disease in animals has shown good results. The study of the effects of viable and inactivated Lactobacillus reuteri (paraprobiotic) can clarify mechanisms of action and enable the commercial use of other preparations, since paraprobiotics would have facilitated production. The aim of this study was to evaluate the effects of viable and inactivated L. reuteri on induced periodontal disease in rats. Thirty-two male rats were divided into four groups: (1) Control group: with periodontal health (n = 8); (2) LIP group: induced periodontitis (n = 8); (3) LIP/PRO group: induced periodontitis and probiotic (n = 8); and (4) LIP/PARAPRO group: periodontitis induced and paraprobiotic (n = 8). Induction of periodontal disease was performed with the insertion of cotton thread around the lower first molars and treatment was performed with viable or heatinactivated L. reuteri daily, 30 days prior to ligation and 14 days later, when euthanasia occurred. The right hemimandible were removed and submitted to radiographic analysis for quantification of the bone support. A computerized microtomography was performed to evaluate the alveolar microarchitecture in the first molar region. The use of paraprobiotic led to a higher percentage of weight gain when compared to other treatments. In the radiographic and microtomographic analyzes, the efficacy of the DP induction was verified, with a significant difference... (Complete abstract click electronic access below) / Mestre

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro de Ciências Agrárias, Programa de Pós-graduação em Ciências dos Alimentos, Florianópolis, 2010 / Made available in DSpace on 2012-10-25T09:03:34Z (GMT). No. of bitstreams: 1 286641.pdf: 985164 bytes, checksum: 2047f4cf8abd0f2d5bc031d595c065ed (MD5) / Em conseqüência ao aumento do interesse dos consumidores por produtos naturais ou minimamente processados, o uso de antimicrobianos naturais pode ser uma alternativa aos métodos utilizados atualmente pela indústria alimentícia. Dessa forma, a aplicação de antimicrobianos naturais vem sendo estudada como alternativa promissora. O microrganismo Lactobacillus reuteri é uma espécie heterofermentativa que reside no trato gastrointestinal, vaginal e oral do homem e outros animais de sangue quente e apresenta a capacidade de produzir antimicrobianos. O presente estudo objetiva avaliar a ação antimicrobiana de L. reuteri contra bactérias de interesse alimentar e estudar o seu controle sobre a microbiota psicrófila de camarão refrigerado. A avaliação da ação antimicrobiana foi realizada pelo método de difusão em agar e pelos métodos Silmultâneo e Deferred em caldo. O estudo do controle da microbiota psicrófila de camarão se deu através da aplicação de uma cultura de L. reuteri e do seu extrato estéril em amostras de camarão que foram mantidas a temperatura de ±7 ºC por 48 horas. L. reuteri apresentou capacidade de inibir o crescimento de Aeromonas hydrophila, Bacillus cereus, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa, Salmonella tiphymurium, Staphylococcus aureus e Vibrio cholerae sendo que a diferença entre a contagem do controle e do teste apresentou diferença estatística. O extrato estéril de L. reuteri foi capaz de reduzir a contagem de microrganismos psicrófilos em 2 ciclos Log, enquanto que a sua cultura manteve a contagem inicial dos microrganismos analisados durante o tempo de armazenamento. L. reuteri foi eficiente no controle de microrganismos psicrófilos no camarão sugerindo o seu uso como bioconservante em alimentos. / As a result of increased consumer interest in natural or minimally processed products, the use of natural antimicrobials may be an alternative to methods currently used in the food industry. Thus, the application of natural antimicrobials has been studied as a promising alternative. The microorganism Lactobacillus reuteri is a heterofermentative species that resides in the gastrointestinal tract, vaginal and oral humans and other warm-blooded animals and has the ability to produce antibiotics. This study aims to evaluate the antimicrobial activity of L. reuteri against bacteria of food interest and study its control over the psychrophilic microbiota of chilled shrimp. The evaluation of antimicrobial activity was performed by agar diffusion method end also by simultaneous and deferred methods in broth. The study of control of the psychrophilic microbiota of shrimp was made through the application L. reuteri culture and sterile extract in the samples of shrimp that were kept at a temperature of ± 7 ° C for 48 hours. L. reuteri showed ability to inhibit the growth of Aeromonas hydrophila, Bacillus cereus, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas peruginosa, Salmonella tiphymurium, Staphylococcus aureus and Vibrio cholera. The sterile extract of L. reuteri was able to reduce the count of psychrophilic microorganisms in two log cycles, whereas the culture maintained the initial count during the storage time. L. reuter was proved effective in controlling microorganisms psychrophiles shrimp suggesting its use as preservative in foods.

Orientador: Letícia Helena Theodoro / Coorientador: Valdir Gouveia Garcia / Banca: Edilson Ervolino / Banca: João Eduardo Gomes Filho / Banca: Guilherme José Pimentel Lopes de Oliveira / Banca: Karina Gonzales Silvério Ruiz / Resumo: Os probióticos (PROs) estão sendo intensamente avaliados na prevenção ou tratamento de doenças da cavidade oral, que estão associadas a uma mudança na composição e atividade microbiana do biofilme e resposta do hospedeiro. O objetivo desse estudo foi avaliar o efeito do Lactobacillus reuteri aplicado localmente ou sistemicamente como coadjuvante ao tratamento periodontal não cirúrgico de raspagem e alisamento radicular (RAR), na periodontite experimentalmente induzida em animais submetidos à quimioterapia com 5-fluorouracil (5-FU). A indução da periodontite experimental (PE) foi realizada com a colocação de um fio de algodão ao redor dos molares inferiores esquerdos de 108 ratos, que permaneceu por 7 dias. Todos os animais receberam o quimioterápico no momento da indução da PE (60mg/kg) e 48 horas após (40mg/kg) e foram separados aleatoriamente em 6 grupos com 18 animais cada um, com os seguintes tratamentos: 5FU: animais tratados com 5-FU; 5FU/RAR: animais tratados com 5-FU, que receberam tratamento com RAR seguido de 1 aplicação de solução fisiológica salina (SS; 0,16mL); Grupo 5FU/SSL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de SS (0,16mL); 5FU/SSS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com SS (0,16mL); 5FU/PL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de PRO (0,16 mL); 5FU/PS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com PRO ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Probiotics (PROs) have been intensively evaluated to prevent or treat oral cavity diseases, associated with a change in the composition and microbial activity of the biofilm and response of the host. The aim of this study was to evaluate the effect of Lactobacillus reuteri applied locally or systemically as a coadjuvant to the non-surgical periodontal treatment of scaling and root planing (SRP) in experimentally induced periodontitis in animals treated with 5-fluorouracil (5-FU). Induction of experimental periodontitis (EP) was performed by placing a cotton thread around the left lower molars of 108 rats, which were kept there for 7 days. All animals underwent chemotherapy twice. They were 60mg/kg at the time they were subjected to ligature induction and again another 40 mg/kg, 48 hours later. They were randomly divided into 6 groups with 18 animals each and they received the following treatments: 5FU: treated animals with 5-FU; 5FU/SRP: 5-FU treated animals receiving SRP treatment followed by physiological saline (SS) solution; 5FU/SSL group: animals treated with 5-FU, who received SRP treatment and local SS applications; 5FU/SSS: animals treated with 5-FU, who received SRP treatment and systemic treatment with SS; 5FU/PL: animals treated with 5-FU, who received SRP treatment and local applications of PRO; 5FU/PS: animals treated with 5-FU, who received SRP treatment and systemic treatment with PRO (0.16ml x 4 days). Six animals from each group were submitted to euthanasia a... (Complete abstract click electronic access below) / Doutor

Die Hemmwirkung des regelmäßigen Konsums probiotischer Lactobacillus reuteri-Stämme auf die Ausprägung oraler Entzündungen ist mittlerweile durch eine ganze Reihe klinischer Interventionsstudien gut belegt. Die allgemeinen Lebensumstände der untersuchten Probanden waren dabei jedoch in der Regel wenig standardisiert, so dass eine mögliche Beeinträchtigung der Validität der Studiendaten durch nicht kontrollierte externe Faktoren wie etwa Lebensstil oder Ernährung bislang nicht ausgeschlossen werden konnte. Daher war es das Ziel dieser prospektiven, randomisierten, doppelt verblindeten und placebokontrollierten Interventionsstudie über einen Beobachtungszeitraum von sechs Wochen die Auswirkungen des täglichen Konsums probiotischer, L. reuteri-haltiger Lutschtabletten auf Parameter der oralen Gesundheit von 72 Besatzungsmitgliedern einer Fregatte der Deutschen Marine zu evaluieren, die während einer Einsatzvorbereitung in See unter weitgehend vergleichbaren Lebens- und Ernährungsbedingungen ihren Dienst versahen. Zu Studienbeginn, sowie nach zwei und sechs Wochen wurden an den Ramfjordzähnen (Zähne 16, 21, 24, 36, 41, 44) der Probanden die Anzahl der auf Sondierung blutenden Zahnfleischtaschen (BoP) als primärem Studien-endpunkt erfasst. Darüber hinaus wurden als sekundäre Endpunkte die Taschensondierungstiefe (PPD), das klinische Attachmentniveau (CAL), der Gingival-Index (GI) und der Plaque Control Record (PCR) aufgezeichnet. Mit Hilfe einer doppelt verblindeten Zuteilungsstrategie wurden die Probanden zufällig der L. reuteri-Gruppe (n=36) oder der Placebogruppe (n=36) zuge-ordnet. Sie erhielten nachfolgend einen für die Studiendauer ausreichenden Vorrat an L. reuteri- oder Placebo-Lutschtabletten mit der Anweisung, diese in den nächsten sechs Wochen zweimal täglich zu konsumieren. 30 Probanden der L. reuteri-Gruppe sowie 32 Probanden der Placebogruppe beendeten die Studie mit vollständig erfassten Datensätzen. Ihre Analyse enthüllte für die L. reuteri-Gruppe einen signifikanten (p<0,001) Rückgang der beobachteten BoP-Mittelwerte von initial 41% (±22 SD) aller erfassten Messstellen auf 10 % (±13 SD) nach sechs Wochen. In der Placebogruppe hingegen kam es während des Beobachtungszeitraums zu einer signifikanten (p=0.05) Zunahme der BoP-Mittelwerte gegenüber der Ausgangssituation von initial 37 % (±20 SD) auf 43 % (±17 SD) am Studienende. Auch bezüglich aller sekundären Endpunkte (PPD, CAL, GI, PCR) konnte in der L. reuteri-Gruppe eine signifikante Verbesserung der oralen Gesundheit zwischen Studienbeginn und Studienende beobachtet werden, während sich wiederum in der Placebo-Gruppe im Beobachtungszeitraum eine statistisch verifizierbare Verschlech¬terung aller erfassten sekundären Endpunkte ergab. Die Ergebnisse dieser unter weitgehend kontrollierten Lebens- und Ernährungsbedingungen durchgeführten Untersuchung belegen, dass der regelmäßigem Konsum probiotischer, L. reuteri-haltiger Lutschtabletten unter den Einsatzbedingungen in See nicht nur eine in der Placebogruppe beobachtete Verschlechterung der oralen Gesundheit verhinderte, sondern diese vielmehr im Vergleich zum Ausgangsbefund signifikant verbesserte. Der adjunktive Konsum L. reuteri-haltiger Lutschtabletten könnte daher eine kostengünstige und einfach zu implementierende Maßnahme darstellen, um einer unter militärischen Einsatzbedingungen häufiger zu beobachtenden Verschlechterung der oralen Gesundheit wirksam vorzubeugen. / The inhibitory effect of the regular consumption of probiotic Lactobacillus reuteri strains on the occurrence of oral inflammations has been established by numerous clinical intervention trials. In general however social ,dietary and environmental conditions of the study subjects were not well controlled, so that so far, it could not be precluded, that the validity of the recorded data may have been possibly impaired by external interference factors like lifestyle and nutrition. Therefore it was the aim of this prospective, randomized, double-blind placebo controlled intervention trial to evaluate the impact of the daily consumption of probiotic L. reuteri-containing lozenges on parameters of oral health in 72 crew members of a frigate of the German Navy over an observation period of 6 weeks during a practicing mission at sea under broadly comparable living and dietary conditions. At baseline, as well as after two and six weeks the number of periodontal pockets showing bleeding on probing (BoP) on the Ramfjord teeth (16, 21, 24, 36, 41, 44) were recorded as the primary study outcome. Additionally pocket probing depth (PPD), periodontal attachment level (PAL), Gingival Index (GI) and Plaque Control Record (PCR) were recorded as secondary study outcomes. Using a computer generated random list study subjects were randomly assigned to either the L. reuteri (n=36) or the placebo group (n=36), provided with a supply of experimental L. reuteri- or placebo lozenges and instructed to consume them twice daily for the next 6 weeks. 30 subjects of the L. reuteri and 32 subjects of the placebo group finished the study with complete data sets. For the L. reuteri group statistical analysis revealed a significant (p<0.001) reduction of the observed percentage of BoP positive sites from 41% (±22 SD) at baseline to 10% (±13) at 6 weeks. In the placebo group by contrast a significant increase (p<0.05) of the percentage of BoP positive sites from 37% (±20 SD) at baseline to 43% (±17%) at 6 weeks was recorded. Also regarding the secondary study outcomes (PPD, CAL, GI, PCR) a significant improvement of all assessed parameters was observed in subjects of the L. reuteri group, while in the placebo group a significant deterioration was recorded. The results of this intervention trial performed under broadly controlled comparable living and dietary conditions revealed, that the regular consumption of probiotic L-reuteri-containing lozenges during a practicing mission at sea not only prevented a deterioration of oral health as documented for the placebo group but even improved the status of oral health when compared to baseline values. The adjunctive consumption of L.reuteri-containing lozenges may therefore present a cost-efficient and easy implementable measure for the prevention of oral health deterioration under military operational conditions.

Proefschr. Rijksuniversiteit Groningen. / Met lit. opg.-Met samenvatting in het Nederlands.

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No. of bitstreams: 1 miessi_dmj_dr_araca_int.pdf: 3518949 bytes, checksum: c91dbdb34520858a3b8383c39b76d5d7 (MD5) Previous issue date: 2018-10-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Os probióticos (PROs) estão sendo intensamente avaliados na prevenção ou tratamento de doenças da cavidade oral, que estão associadas a uma mudança na composição e atividade microbiana do biofilme e resposta do hospedeiro. O objetivo desse estudo foi avaliar o efeito do Lactobacillus reuteri aplicado localmente ou sistemicamente como coadjuvante ao tratamento periodontal não cirúrgico de raspagem e alisamento radicular (RAR), na periodontite experimentalmente induzida em animais submetidos à quimioterapia com 5-fluorouracil (5-FU). A indução da periodontite experimental (PE) foi realizada com a colocação de um fio de algodão ao redor dos molares inferiores esquerdos de 108 ratos, que permaneceu por 7 dias. Todos os animais receberam o quimioterápico no momento da indução da PE (60mg/kg) e 48 horas após (40mg/kg) e foram separados aleatoriamente em 6 grupos com 18 animais cada um, com os seguintes tratamentos: 5FU: animais tratados com 5-FU; 5FU/RAR: animais tratados com 5-FU, que receberam tratamento com RAR seguido de 1 aplicação de solução fisiológica salina (SS; 0,16mL); Grupo 5FU/SSL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de SS (0,16mL); 5FU/SSS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com SS (0,16mL); 5FU/PL: animais tratados com 5-FU, que receberam tratamento com RAR e 4 aplicações locais de PRO (0,16 mL); 5FU/PS: animais tratados com 5-FU, que receberam tratamento com RAR e 4 gavagens com PRO (0,16mL). 6 animais de cada grupo foram submetidos à eutanásia aos 7, 15 e 30 dias após os tratamentos. A área da furca dos molares foi submetida às análises histopatológica, histométrica e dos padrões de imunomarcação para TRAP, PCNA, RANKL, OPG, OCN e TGF-β1. Os dados obtidos foram submetidos à análise estatística (α=5%). O grupo 5FU/PS apresentou melhora no processo inflamatório em todos os períodos. Houve maior perda óssea alveolar (PO) nos espécimes do grupo 5FU/SSS comparado ao grupo 5FU/SSL (p˂0,01) no período de 7 dias de avaliação. No período de 15 dias os espécimes dos grupos 5FU/SSL e 5FU/SSS demonstraram maior PO comparado aos grupos 5FU (p˂0,05) e 5FU/PS (p˂0,05). Aos 30 dias a PO mostrou-se maior nos espécimes do grupo 5FU/PL comparado aos grupos 5FU/RAR (p˂0,05), 5FU/SSS (p˂0,05) e 5FU/PS (p˂0,05). Na análise intragrupo houve menor PO no grupo 5FU/RAR e 5FU/PS aos 30 dias em relação aos 7 dias (p˂0,05). Na imunomarcação de células TRAP-positivas não foi evidenciada diferença significante entre os grupos e períodos; houve maior imunomarcação de células PCNA-positivas aos 7 dias no grupo 5FU/RAR comparado ao 5FU/SSS. Prevaleceu baixo padrão de imunomarcação de TGF-β1 e OCN nos grupos 5FU, 5FU/RAR, 5FU/SSL e 5FU/SSS em todos os períodos. O grupo 5FU/PS apresentou aos 7 dias um moderado padrão de imunomarcação e aos 15 e 30 dias um alto padrão de imunomarcação de TGF-β1 e OCN; o grupo 5FU/PL apresentou um moderado padrão de imunomarcação de TGF-β1 e OCN aos 7,15 e 30 dias. Na análise de RANKL prevaleceu um alto padrão de imunomarcação nos grupos 5FU, 5FU/RAR, 5FU/SSL, 5FU/SSS em todos os períodos e o grupo 5FU/PS apresentou aos 7 e 15 dias um moderado padrão de imunomarcação e aos 30 dias predominou um padrão de imunomarcação que variou do moderado ao baixo. Prevaleceu um padrão baixo de imunomarcação em todos os grupos experimentais e períodos, com exceção dos grupos 5FU/PL e 5FU/PS aos 30 dias (moderado padrão de imunomarcação) na análise de OPG. Diante dos resultados obtidos e com a metodologia empregada pode ser concluído que o uso sistêmico do Lactobacillus reuteri promoveu redução da inflamação e beneficiou o processo de reparação dos tecidos periodontais, porém não foi capaz de reduzir a PO na região de furca, demonstrando ser uma terapia periodontal coadjuvante promissora em animais submetidos à quimioterapia com 5-FU. / Probiotics (PROs) have been intensively evaluated to prevent or treat oral cavity diseases, associated with a change in the composition and microbial activity of the biofilm and response of the host. The aim of this study was to evaluate the effect of Lactobacillus reuteri applied locally or systemically as a coadjuvant to the non-surgical periodontal treatment of scaling and root planing (SRP) in experimentally induced periodontitis in animals treated with 5-fluorouracil (5-FU). Induction of experimental periodontitis (EP) was performed by placing a cotton thread around the left lower molars of 108 rats, which were kept there for 7 days. All animals underwent chemotherapy twice. They were 60mg/kg at the time they were subjected to ligature induction and again another 40 mg/kg, 48 hours later. They were randomly divided into 6 groups with 18 animals each and they received the following treatments: 5FU: treated animals with 5-FU; 5FU/SRP: 5-FU treated animals receiving SRP treatment followed by physiological saline (SS) solution; 5FU/SSL group: animals treated with 5-FU, who received SRP treatment and local SS applications; 5FU/SSS: animals treated with 5-FU, who received SRP treatment and systemic treatment with SS; 5FU/PL: animals treated with 5-FU, who received SRP treatment and local applications of PRO; 5FU/PS: animals treated with 5-FU, who received SRP treatment and systemic treatment with PRO (0.16ml x 4 days). Six animals from each group were submitted to euthanasia at 7, 15 and 30 days after treatments. The area of the molar furcation was submitted to histopathological, histometric and immuno-labeling analysis for TRAP, PCNA, RANKL, OPG, OCN and TGF-β1. The data were submitted to statistical analysis (α = 5%). There was greater bone loss (BL) in the 5FU/SSS group compared to the 5FU/SSL group (p˂0.01) in the 7-day evaluation period. In the 15-day period, specimens from the 5FU/SSL and 5FU/SSS groups showed a higher BL compared to the 5FU (p˂0.05) and 5FU/PS (p˂0.05) groups. At 30 days the BL was higher in the 5FU/PL group compared to the 5FU/SRP (p˂0.05), 5FU/SSS (p˂0.05) and 5FU/PS groups (p˂0.05). In the intragroup analysis there was a lower BL in the 5FU/SRP group and 5FU/PS at 30 days compared to 7 days (p˂0.05).In the immunostaining of TRAP-positive cells no significant difference between groups and periods was evidenced; there was greater immunolabeling of PCNA-positive cells at 7 days in the 5FU/SRP group compared to 5FU/SSS. Low prevalence of TGF-β1 and OCN immunostaining in the 5FU, 5FU/RAR, 5FU/SSL and 5FU/SSS groups prevailed at all periods. The 5FU/PS group presented at 7 days a moderate pattern of immunostaining and at 15 and 30 days a high standard of immunoblotting of TGF-β1 and OCN; the 5FU/PL group presented a moderate pattern of TGF-β1 and OCN immunoregulation at 7, 15 and 30 days; for RANKL a high standard of immunostaining in the 5FU, 5FU/SRP, 5FU/SSL, 5FU/SSS groups prevailed at all periods, 5FU/PL prevailed at 7 days a high standard of immunostaining and a moderate immunostaining pattern at 15 and 30 days and 5FU/PS showed a moderate immunostaining pattern at 7 and 15 days, and at 30 days a predominance of the immunoblot pattern varied from moderate to low; for OPG a low pattern of immunoblotting prevailed in all experimental groups and periods, except for 5FU/PL and 5FU/PS at 30 days that presented a moderate pattern of immunostaining. Based on the results obtained in this study and the methodology used to work on it, it was concluded that the use of Lactobacillus reuteri promoted reduction of inflamation and benefited the process of repairing periodontal tissues, but it was not able to reduce the BL in the furca region, proving to be a therapy promising adjuvant periodontal in animals submitted to 5-FU chemotherapy. / Capes

The crustacean Artemia franciscana represents a potential experimental model for human studies. The aim of the project was to assess whether Artemia could be used as a model for investigation of probiotic properties. For all the experiments, Artemia cysts were rehydrated, decapsulated and allowed to hatch for 28 hours before the developed larvae (nauplii) were challenged and/or treated with probiotics; measurement of survival rate occurred after further 24 and 48 hours. Interestingly, administration of L. reuteri 17938 during an initial study showed that the probiotic rescued Artemia from spontaneous death occurring in the absence of any established and chosen challenge in a dose-dependent manner. Assuming that Artemia death was related to some kind of stress, the mode of action by means of which L. reuteri provided protective effects was investigated. Heat-treated and bead-beaten L. reuteri positively affected Artemia survival rate to a greater extent than live L. reuteri, with the highest improvement occurring during administration of bead-beaten L. reuteri. In particular, administration of different fractions of the bead-beaten L. reuteri cultures to Artemia allowed for identification of the molecule responsible for amelioration of Artemia fitness in bacterial components ranging between 3 and 300 kDa in weight. Supernatants obtained from exudates of plate-grown bacteria instead revealed itself to be lethal when added in amounts corresponding to 10⁷ CFU/ml of the bacterial culture or higher. Further studies on a genetic and immunological level on Artemia are required in order to elucidate the complex interactions establishing among the host, the probiotic and the environment. However, based on the data gathered, it is possible to suggest that Artemia can be used as a potential chronic stress model, for example as aging or starvation models. Besides this, L. reuteri shows to provide an overall protective effect against this stress, although differences in the extent of the protection were observed according to the growth protocol and the killing mode of bacterial cultures. Also, the lethality of the supernatants demonstrates once again that the probiotic activity is the result of multiple and intertwined biological pathways, sometimes in contrast to one another.

Background: Probiotic bacteria have been used for centuries to obtain a better health and the majority of publications have focused on the gut health. More recent studies do also investigate if the bacteria could have any effect in the oral microflora. Since probiotic products are becoming more common on the market, it is interesting to investigate if these probiotic bacteria exist in the oral cavity and if they exert any therapeutic effect on oral diseases. The aim of this study was to determine an uncomplicated method capable of measuring one bacterium associated with caries prophylactic properties. Methods: Saliva from three test subjects were collected before and after chewing on a tablet containing Lactobacillus reuteri. DNA from each saliva sample was extracted using FTA™ elute cards and amplified with a PCR. Saliva samples were cultured on Rogosa agar for comparison. Saliva after chewing was diluted for determination of detection level using Rogosa agar counts as standard. Amplified samples were analysed from stained electrophoresis gels.Results: The PCR method could detect Lactobacillus reuteri in saliva if the content was 250 CFU/mL or higher. An increase in CFU/mL in saliva after chewing can be observed. Saliva could before chewing show in two out of three test subjects no amplifiable DNA whilst after chewing all did.Conclusions: A method considered uncomplicated that is capable of detecting Lactobacillus reuteri in saliva has been developed. Detection level was 250 CFU/mL.

Introduktion. Probiotiska bakterier är erkända som behandling av sjukdomar i tarmen och probiotiska produkter ökar på marknaden. För att kunna utvärdera effekterna i munhålan är det av intresse att undersöka om dessa bakterier, tillsammans med ökad tillgänglighet, utvecklas till en del av den naturliga orala mikrofloran.Syfte. Syftet med studien var att med konventionell PCR-teknik undersöka och jämföra förekomsten av den probiotiska bakterien Lactobacillus reuteri i saliv insamlat från två olika populationer. Syftet var också att fastställa en detektionsnivå för den använda metoden och att bekräfta metodens förmåga att differentiera mellan L. reuteri och andra arter av lactobaciller.Material och Metod. Salivprover samlades in från två olika populationer från Sverige och Danmark. Proverna undersöktes för förekomst av L. reuteri genom PCR-teknik med primer specifik för L. reuteri.Resultat. L. reuteri detekterades i 27 % av salivproverna från båda populationer. Detektionsnivån fastställdes till 103 CFU/mL saliv och metoden hade förmåga att differentiera mellan L. reuteri och andra arter av lactobaciller.Konklusion. Metoden var specifik för detektion av L. reuteri med en detektionsnivå på 103 CFU/mL saliv. Resultaten indikerar att L. reuteri kan kolonisera munhålan hos vissa individer. De två populationerna visade ingen skillnad i prevalens. Detta kan indikera att en förmodad ökad tillgänglighet av produkter innehållande L. reuteri inte har ökat prevalensen över tid, eller att individer skiljer sig i förmågan att koloniseras. / Introduction. Probiotic bacteria are acknowledged as treatment for intestinal diseases and probiotics are increasing on the market. To properly evaluate the effects in the oral cavity it is of interest to investigate if these bacteria, along with increased availability, are becoming a natural part of the oral microflora.Aim. The aim of this study was to investigate and compare the presence of the probiotic bacteria Lactobacillus reuteri in saliva collected from two different populations using conventional PCR method. The aim was also to determine a detection level for the method used and confirm the methods ability to differentiate between L. reuteri and other lactobacillus spp.Material and Method. Saliva samples were collected from two different populations from Sweden and Denmark. Samples were tested for the presence of L. reuteri using PCR-technique with primer specific to L. reuteri.Results. L. reuteri was detected in 27 % of the saliva samples of both populations. The detection level was set to 103 CFU/mL saliva and the method was able to differentiate between L. reuteri and other lactobacillus spp.Conclusion. The method used was specific for detection of L. reuteri with a detection level of 103 CFU/mL saliva. Results indicate that L. reuteri is able to colonize the oral cavity in some individuals. The two populations showed no difference in prevalence. This might indicate that an assumed increased availability of L. reuteri in consumer probiotic products over time has not increased the prevalence, or that individuals differ in their ability to be colonized.

Orientador: Ana Lia Anbinder / Coorientadora: Liliana Scorzoni / Banca: Cristiane Yumi Koga Ito / Banca: Victor Angelo Martins Montalli / Resumo: A doença periodontal, afecção crônica inflamatória multifatorial, está entre as principais doenças bucais que afetam a população mundial. Entre as bactérias associadas à periodontite, estão Fusobacterium nucleatum e Aggregactibacter actinomycetemcomitans. Novas terapias adjuntas ao tratamento convencional têm sido propostas para a doença periodontal, entre elas o uso de probióticos. Porém, seu uso não está isento de riscos, e uma alternativa para minimizá-los é inativar os micro-organismos, mantendo suas propriedades benéficas, o que ocorre com os chamados paraprobióticos. Desse modo, são objetivos deste estudo avaliar os efeitos antimicrobianos de L. reuteri vivo, inativado pelo calor e seus produtos sobre F. nucleatum, A. actinomycetemcomitans e sobre as bactérias comensais, Streptococcus mitis e Streptococcus salivarius, além de estudar os efeitos da interação das preparações e periodontopatógenos em modelo de invertebrado. A atividade antimicrobiana in vitro foi avaliada associando-se as bactérias patogênicas ou comensais a L. reuteri vivo, inativado ou sobrenadante. Após interação, as bactérias foram cultivadas em meio seletivo para contagem de unidades formadoras de colônias (UFC). No estudo em Galleria mellonella, após a infecção com as bactérias patogênicas e as preparações de L. reuteri, foi avaliada a curva de sobrevivência e densidade hemocitária. Os dados foram analisados com o teste estatístico apropriado, ao nível de 5%. Após interação bacteriana in vitro, S. sa... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Periodontal disease, a chronic multifactorial inflammatory disease, is among the major oral diseases that affects the worldwide population. Among the bacteria associated with periodontitis are Fusobacterium nucleatum and Aggregactibacter actinomycetemcomitans. New therapies have been proposed for periodontal disease as adjunct to conventional treatment, including the use of probiotics. However, their use isn't risk-free and an alternative to that is the inactivation of the microorganisms, maintaining their beneficial properties, which occurs with the paraprobiotics. Thus, the objective of this study is to evaluate the antimicrobial effects of living and heat-killed L. reuteri, and its products on F. nucleatum, A. actinomycetemcomitans and on the commensal bacteria, Streptococcus mitis and Streptococcus salivarius, as well as to study the interaction of the preparations and periodontopathogens in an invertebrate model. In vitro antimicrobial activity was evaluated by associating the pathogenic or commensal bacteria to live, heat killed or L. reuteri supernatant. After interaction, the bacteria were cultured in a selective medium for colony-forming unit (CFU) count. In the study with Galleria mellonella, after infection with pathogenic bacteria and L. reuteri preparations, the survival curve and hemocyte density were evaluated. The data were analyzed with the appropriate statistical test at the 5% level. After bacterial interaction in vitro, S. salivarius reduced the number of ... (Complete abstract click electronic access below) / Mestre