A pele dos anfíbios, assim como de outros animais, atua como primeira proteção contra agentes patogênicos. A comunidade microbiológica ali residente é composta de algumas espécies de bactérias, e estas, possuem ação antifúngica contra patógenos conhecidos, inclusive Batrachochytrium dendrobatidis (Bd), o suposto agente principal de declínios de populações de anfíbios em diversas partes do mundo. Uma vez que as variáveis químicas e físicas de um ecossistema influenciam o crescimento, sobrevivência e atividade metabólica dos microorganismos, a microbiota cutânea que atua como barreira de proteção nos anfíbios contra agente infecciosos, provavelmente é afetada quando determinados parâmetros ecofisiológicos são alterados em ambientes florestais fragmentados, modulando assim a vulnerabilidade das populações de anfíbios aos agentes patogênicos. Nossa pesquisa esteve focada na caracterização das comunidades microbianas residentes da pele dos anfíbios em dois contextos de paisagem: fragmento e área contínua. Os parâmetros utilizados para essas análises foram a densidade microbiana e a riqueza de morfotipos de colônias bacterianas. O potencial inibitório do crescimento de patógenos também foi testado em ensaios do tipo cross-strak. As diferenças de densidade e riqueza microbiana entre as paisagens e a presença de táxons típicos de ambiente, apontam para o ambiente como um componente importante na determinação dos perfis das comunidades microbianas dos anfíbios estudados. Essas mudanças são muito provavelmente conseqüências, mas para o entendimento da extensão e natureza de tais conseqüências são necessários estudos adicionais. / The skin of amphibians, as well as that of other animals, acts as a first protection barrier against pathogens. The microbial community resident in the amphibian skin is composed of some species of bacteria that may have antibacterial or antifungal action against known pathogens, including Batrachochytrium dendrobatidis, the alleged principal agent Tleading to declines of amphibian populations around the world. Because the chemical and physical variables of the landscape influence the growth, survival and metabolic activity of microorganisms, the function of skin as a protective barrier against infectious agents in amphibians, is likely affected by parameters that are altered in fragmented forest habitats. Thus, it is important to understand how environmental conditions affect the skin microbiota of amphibians, and the possible induced changes on vulnerability of amphibians to pathogens. Our research aimed to characterize the microbial communities living skin of amphibians in two contexts of landscape: fragment and continuous area. The parameters used for this analysis were density and richness of microbial morphotypes of bacterial colonies. The potential inhibition of pathogen growth was also evaluated using a cross-streak test, and some taxa in these communities were identified using international protocols. The observed differences in microbial density and richness across landscapes, and the presence of bacterial taxa typical of given environments, point out to the role of environmental change as an important component determining the profiles of microbial communities living on the skin of amphibians. These changes are very likely consequential, but understanding the scope and nature of consequences require additional study.

Incorporation of probiotic cultures in products in order to replenish or supplement the normal gastrointestinal microflora is a well known and accepted practice. However survival of these cultures is a problem due to a number of reasons including effects of storage conditions. Various researchers from different countries around the world have reported probiotic product instability. Microencapsulation has been used in an attempt to solve this problem. However, most methods involve the use of organic solvents which is not ideal because their toxicity may cause destruction of the microbial cells. A novel encapsulation method for probiotics, which excludes the use of organic solvents, was developed by the Council for Scientific and Industrial Research (CSIR) (US Patent Application no. 20050112205). This thesis investigated the efficiency/potential of this new method for increasing stability of sensitive probiotic cultures, specifically bifidobacteria. Early studies using both culture dependent and culture independent techniques showed reduced numbers of viable cultures in probiotic products, mainly yoghurts, from all around the world. These results were confirmed in this study for similar products sold in South Africa. Most of the product labels did not specify viable numbers of probiotics nor the identity (genus and species names) of the microorganisms incorporated. Successful encapsulation of bifidobacteria was achieved using the CSIR patented method. Complete encapsulation was indicated by absence of cells on surfaces of the encapsulated particles and production of a product with an acceptable particle size distribution was obtained. It was also demonstrated that the encapsulation process produced no visible morphological changes to the bacterial cells nor did it have a negative effect on cell viability over time. The potential of interpolymer complex formation in scCO2 for the encapsulation of sensitive probiotic cultures was demonstrated for the first time. Once ingested, probiotic cultures are exposed to unfavourable acidic conditions in the upper gastrointestinal tract. It is desired that these cultures be protected from this in order to increase the viability of the probiotics for efficient colonization. Interpolymer complex encapsulated B. longum Bb-46 cells were therefore exposed to simulated gastric fluid (SGF) and subsequently to simulated intestinal fluid (SIF). It was found that the interpolymer complex protected bifidobacteria from gastric acidity, displaying pH-responsive release properties, with little to no release in SGF and substantial release in SIF. Thus the interpolymer complex demonstrated desirable characteristics retaining the encapsulated bacteria inside when conditions were unfavourable and only releasing them under favourable conditions. Survival was improved by the incorporation of glyceryl monostearate (GMS) in the matrix and by use of gelatine capsules. Protection efficiency of the interpolymer matrix was better when higher loading of GMS was used. Use of polycaprolactone (PCL) as an alternative to poly (vinylpyrrolidone) (PVP) and incorporation of ethylene oxide-propylene oxide triblock copolymer (PEO-PPO-PEO) affected the interpolymer complex negatively, rendering it swellable in the low pH environment exposing the bifidobacteria to gastric acidity. The use of beeswax seemed to have a more protective effect though results were inconclusive. Probiotic cultures must also remain viable in products during storage. Encapsulated bacteria were either harvested from the reactor after 2 h of equilibration followed by depressurization, and then ground to a fine powder or after 2 h of equilibration the liquefied product was sprayed through a capillary tube with a heated nozzle at the end, into the product chamber. Encapsulated bacteria were stored in either sterile plastic bags or glass bottles under different conditions and then viable counts were determined over time. Survival of bacteria was generally better when the products were stored in glass bottles than in plastic bags. Bacteria encapsulated in an interpolymer complex formed between PVP and vinyl acetate-crotonic acid copolymer (VA-CA), (PVP:VA-CA) survived better than non-encapsulated bacteria under all storage conditions when the product was recovered from the reaction chamber. When the product was recovered from the product chamber, numbers of viable non-encapsulated bacteria were higher than the encapsulated bacteria for all interpolymer complex formulations. This was probably due to some exposure to high shear during spraying into the product chamber. The interpolymer complex between PCL and VA-CA i.e. PCL:VA-CA seemed weaker than the PVP:VA-CA nterpolymer complex as viable counts of bacteria released from it were lower than those from the latter complex. Addition of PEO-PPO-PEO to both the PVP:VA-CA and PCL:VA-CA complexes decreased the protection efficiency. However, results indicated that sufficient release of encapsulated bacteria from the interpolymer complexes was obtained when the encapsulated material was incubated in SIF rather than in Ringer’s solution. When SIF was used for release of encapsulated bacteria, the shelf life of B. longum Bb-46 was doubled. Encapsulation in an interpolymer complex therefore provided protection for encapsulated cells and thus has potential for improving shelf life of probiotic cultures in products. Further studies will investigate the effects of encapsulating probiotics together with prebiotics in the interpolymer complex as well as effects of encapsulating combinations of different probiotic strains together, both on survival in simulated gastrointestinal tract and during storage. The unique particles produced using the patented encapsulation technique increased the stability of probiotic cultures. This technique may find significant application in industries manufacturing probiotic products, especially food and pharmaceuticals, thereby improving the well being of consumers. / Thesis (PhD(Microbiology))--University of Pretoria, 2008. / Microbiology and Plant Pathology / PhD / unrestricted

Serotonin (5-hydroxytryptamine, 5-HT) has been shown to activate carbohydrate metabolism in adult female Ascaris suum. Serotonin may be either absorbed directly from the environment or synthesized de novo from the absorbed L-tryptophan in adult female A. suum. The enzymes necessary for the synthesis of 5-HT have been identified in both intestine and muscle tissues. The serotonin absorbed from the environment is obtained either from the host's gastrointestinal contents or from the 5-HT producing bacteria in the intestine of A. suum. Numerous 5-HT producing bacteria were identified in the intestinal microflora. The physiological contributions of 5-HT producing bacteria to the 5-HT level, turnover and binding sites in the intestinal tissue of A. suum were investigated.

No description available.

No description available.

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L’esca est une maladie de dépérissement du bois de la vigne conduisant à la mort des ceps. Actuellement le vignoble mondial est atteint, et au niveau français, cette maladie ne cesse de progresser. Ainsi, 8% des ceps dans le Jura et 4,5% dans la région de Bordeaux manifestent des symptômes d’esca, selon les parcelles des chiffres beaucoup plus élevés sont obtenus, certains cépages sont aussi beaucoup plus sensibles que d’autres. Plusieurs champignons seraient impliqués dans l’esca mais leur rôle ainsi que la détermination de la microflore responsable de cette maladie est encore sujette à interrogation. Dans ce contexte, l’objectif de cette thèse a été de caractériser et de comparer les microflores fongiques et bactériennes colonisant le bois de ceps de vigne ayant exprimé ou non des symptômes foliaires d’esca. Dans un premier temps, nous avons prélevé des ceps (cultivar Cabernet Sauvignon) relativement jeunes (10 ans d’âge) car ils présentaient l’intérêt d’être peu dégradés au niveau du bois du tronc, les symptômes foliaires étant associés à la présence d’amadou (une nécrose typique de l’esca) uniquement dans les bras. Une grande diversité dans les communautés fongiques (674 OTUs) et bactériennes (222 OTUs) colonisant le bois a été observée. Cette diversité est plus importante dans le bois sain de la vigne que dans celui partiellement ou totalement nécrosé. Les techniques utilisées, i.e. isolement/séquençage de souches, empreinte moléculaire (Single Strand Conformation Polymorphism, SSCP) et pyroséquençage 454, ont montré que les communautés bactériennes ou fongiques étaient différentes dans les tissus dégradés comparés à ceux qui ne l’étaient pas. Des changements de microflores en fonction du temps (expérimentation durant 1 année) ont aussi été observés. D’une façon générale, les espèces de champignons impliquées dans l’esca sont déjà présentes dans le bois apparemment sain de ceps esca-foliaires symptomatiques mais aussi des asymptomatiques. Il n’a pas été possible de différencier ces 2 types de microflores au niveau du bois sain des plants, cette différentiation se faisant au niveau des nécroses, qui sont plus abondantes dans les ceps esca-symptomatiques. Pour la première fois nous avons montré que des communautés bactériennes spécifiques étaient associées à l’esca, leurs aptitudes trophiques étant différentes selon les tissus où elles étaient prélevées. Les espèces isolées suggèrent que certaines pourraient avoir un rôle dans la protection du végétal, d’autres dans la dégradation des structures du bois, e.g. de la lignine, préparant ainsi le terrain aux champignons dégradateurs des tissus ligneux, déjà présents à l’intérieur des ceps. Nous avons aussi étudiés des ceps plus âgés (cultivar Baco blanc), de 42 et 58 ans, qui avaient un rendement acceptable et n’avaient pas manifesté de symptômes d’esca ou eutypiose (une autre maladie du bois) l’année du prélèvement. Au niveau des tissus fonctionnels du bois, les communautés fongiques étaient caractéristiques de plants atteints par l’eutypiose (ceps de 42 ans) ou de l’esca (ceux de 58 ans). La non expression par les ceps de ces 2 maladies pourrait cependant être associée à la forte présence de champignons mycoparasites et protecteur du végétal, comme Trichoderma spp., dans ces tissus fonctionnels. Les interactions au sein des communautés fongiques créant un équilibre où le pathogène ne se développerait pas de façon extensive. Les caractéristiques du Baco blanc, un hybride, moins sensible à certaines maladies de la vigne, pourrait aussi expliquer ce résultat. Ainsi la présence d’une microflore bénéfique naturellement présente dans le bois des ceps associée à des plants ayant une tolérance à ces maladies pourrait ouvrir de nouvelles perspectives pour lutter l’esca, voire l’eutypiose, pour lesquelles aucun moyen de protection n’existe aujourd’hui. / Esca is a Grapevine Trunk Disease (GTD) that induces a decline in grapevine vigour that generally leads up with the death of the plants. Nowadays, vineyards worldwide are attacked by esca and, in France this disease increases steadily. In the Jura, 8% of the grapevines are esca-foliar symptomatic and approximately 4.5% in the Bordeaux region. However, some vineyards are more severely attacked by esca, and certain cultivars are more susceptible than others. Although several pathogenic fungi are associated with esca, their individual roles and their interaction with other microorganisms for the esca have still to be determined. In this context, the objective of the present PhD study is to characterize and compare the bacterial and fungal microflora that colonize the wood tissues of esca-foliar symptomatic and asymptomatic grapevines. First, we sampled young (10 year-old) grapevines (Cabernet Sauvignon cultivar) because they had only few necroses in the trunk and white-rot (also called amadou) was only present in the cordons of symptomatic plants. Great diversity in the fungal (674 OTUs) and bacterial (222 OTUs) communities was observed. This diversity was higher in the apparently healthy wood than in the partially or totally necrotic wood tissues. The methods used isolation/sequencing of microbial strains, a molecular fingerprinting method (Single Strand Conformation Polymorphism, SSCP) and 454 pyrosequencing showed that the fungal and bacterial communities of the necrotic and healthy wood tissues were different. Changes in the microflora over time (over a one-year period) have been observed. Fungal species involved in esca are already present in the apparently healthy wood of esca-foliar symptomatic plants but also in the asymptomatic ones. It was not possible to differentiate these 2 microflora. Only microflora from the necroses differed from those of the healthy wood with these necroses being more developed in the esca-foliar symptomatic grapevines. For the first time, we were able to determine that specific bacterial communities are associated with esca. Depending on the wood tissues, different types of bacteria were isolated, with different trophic behaviour. Two roles could be assigned to the species isolated from the various wood tissues: (i) a positive role, due to the biocontrol potential that many species have; (ii) a negative one, by predisposing the wood of grapevines to fungal attacks. We also studied, old (42 and 58 year-old) grapevines of the cultivar, Baco blanc, that produced regular harvests. The plants had no expressed foliar symptoms of esca or eutypa dieback during the sampling year. Many plant pathogens colonized the functional wood tissues, but in 58 year-old plants they were associated with esca, and in 42 year-old plants, with eutypa dieback. The absence of GTDs expression could be linked to the numerous plant protectant mycoparasites, such as Trichoderma spp., that colonized the functional wood tissues. Interactions between species within the fungal communities may create a balance that is unfavourable to the development of the pathogens. The use of Baco blanc, a hybrid less susceptible to certain grapevine diseases could also explain this result. So, because no means of protection are currently available, the combination of beneficial microflora within the garpevine wood tissues with plants that are tolerant to esca, or even eutypa dieback, could be helpful to control those diseases.

Submitted by Michele Mologni (mologni@unoeste.br) on 2019-01-29T18:40:59Z No. of bitstreams: 1 Sandra Cristina Genaro.pdf: 1632390 bytes, checksum: 89f6ec224705d20a5ce967976b70a4c3 (MD5) / Made available in DSpace on 2019-01-29T18:40:59Z (GMT). No. of bitstreams: 1 Sandra Cristina Genaro.pdf: 1632390 bytes, checksum: 89f6ec224705d20a5ce967976b70a4c3 (MD5) Previous issue date: 2018-11-22 / Among the existing types, colorectal cancer (CRC) affects approximately one million people per year, considered the second most common cause of death among women and the third most prevalent in men. Risk factors are genetic syndromes; inflammatory bowel diseases; family history; sedentary lifestyle; obesity, low fiber, high saturated fats, smoked foods or built-in food, excess red meat (> 300g/week), preparation mode in high temperatures and on the ember; medications; smoking, and excessive alcohol. These factors alter the intestinal microbiome which is colonized by pathogenic bacteria capable of provoking a local inflammatory response that, in chronic cases, activates carcinogenic components. Probiotics have increasingly attracted the attention of researchers in order to understand their action in the intestinal microbiota, aiding in the prevention and treatment of colorectal cancer. The objective of this study was to evaluate the effect of a probiotic In aggressiveness of the chemically induced colorectal tumor in rats. Twenty-five male Fisher 344 rats, 250 g, receiving ration and water ad libitum, were randomly divided into 5 groups (5 rats/group): GControl, without treatment; GTumor, tumor induction; GTumor + 5FU, tumor induction, 5-fluorouracil applied; GTumor + prob, tumor induction, supplemented with probiotic; GTumor + 5-FU + prob, tumor induction, applied 5-fluorouracil, supplemented with probiotic. For tumor induction, the animals received four intraperitoneal injections of the carcinogen 1.2-dimethylhidrazine (DMH) at the dose of 20 mg/kg body weight, being two applications per week, for four consecutive week. A 15-day interval was given and DMH applications were repeated for another four week. After 5 weeks of the last dose of the carcinogen, the treatment was initiated for ten consecutive weeks, applying a weekly dose of 15 mg/kg body weight of 5-fluorouracil, Intraperitoneal route and commercial probiotic containing Lactobacillus and Bifidobacterium at the dose of 1x109 UFC, administered by gavage, daily. Datas were analyzed by the analysis of variance One Way and the averages compared by the test of Dunnett. Used Software Statistical GraphPad Prism. The histopathologic analyses evaluated by the Chi-square ratio test. It was considered type-I error of 5% as statistically significant. Compared to the GTumor, the GTumor + prob (P < 0,0373) and GTumor + 5-FU + prob (P < 0,0003) showed attenuating effect on the aggressiveness of the colorectal tumor, with a reduction in the count of Aberrant Crypts Foci; and lower percentage of malignant neoplastic lesions in the GTumor + prob (40% of low-grade tubular adenoma, 40% of carcinoma in situ, 20% of low-grade adenocarcinoma) and GTumor + 5-FU + prob (40% of low-grade tubular adenoma and 60% of carcinoma in situ). The suplementation with probiotic has the potential to decrease the formation of aberrant crypts and mitigate the progression of tumor malignancy, potentializing the antitumor effect of 5-fluorouracil chemotherapy in the colic segments. / O câncer colorretal (CCR) acomete aproximadamente um milhão de pessoas por ano, considerado a segunda causa de morte mais comum entre mulheres e a terceira mais prevalente em homens. Os fatores de risco incluem as síndromes genéticas; doenças inflamatórias intestinais; história familiar; sedentarismo; obesidade, alimentação pobre em fibras, rica em gorduras saturadas, alimentos defumados ou embutidos, carne vermelha em excesso (>300g/sem), modo de preparação em altas temperaturas e na brasa; medicamentos; tabagismo e bebida alcoólica em excesso. Esses fatores levam a alteração da microbiota intestinal a qual é colonizada por bactérias patogênicas capazes de provocar uma resposta inflamatória local que, em casos crônicos, ativam componentes cancerígenos. Os probióticos têm atraído cada vez mais a atenção de pesquisadores com o intuito de compreender a sua ação na microbiota intestinal, auxiliando na prevenção e tratamento do câncer colorretal. O objetivo desse estudo foi avaliar o efeito de um probiótico na agressividade do tumor colorretal induzido quimicamente em ratos. Vinte e cinco ratos machos Fisher 344, 250 g, recebendo ração e água ad libitum, foram divididos aleatoriamente em 5 grupos (5 ratos/grupo): GControle, sem tratamento; GTumor, indução do tumor; GTumor+5FU, indução do tumor, aplicado 5-Fluorouracil; GTumor+Prob, indução do tumor, suplementado com probiótico; GTumor+5-FU+Prob, indução do tumor, aplicado 5-Fluorouracil, suplementado com probiótico. Para indução do tumor colorretal, os animais receberam quatro injeções intraperitoneais do carcinógeno 1,2-dimetilhidrazina (DMH) na dose de 20 mg/kg de peso corporal, sendo duas aplicações por semana, durante quatro semanas consecutivas. Deu-se um intervalo de 15 dias e as aplicações de DMH foram repetidas por mais quatro semanas. Após 5 semanas da última dose do carcinógeno, iniciou-se o tratamento por dez semanas consecutivas, com 5-Fluorouracil: uma dose de 15 mg/kg por semana, via intraperitoneal e probiótico comercial: 1x109 UFC, diariamente, por gavagem. Os dados foram analisados pela Análise de Variância One Way e as médias comparadas pelo teste de Dunnett. Utilizado software estatístico GraphPad Prism. As análises histopatológicas avaliadas pelo teste de proporção Qui-quadrado. Foi considerado erro tipo-I de 5% como estatisticamente significante. Comparados com o GTumor, o GTumor+Prob (p<0,0373) e GTumor+5-FU+Prob (p<0,0003) exibiram efeito atenuante na agressividade do tumor colorretal obervando redução na contagem de Focos de Criptas Aberrantes; e menor porcentagem de lesões neoplásicas malignas no GTumor+Prob (40% de adenoma tubular de baixo grau, 40% de carcinoma in situ, 20% de adenocarcinoma de baixo grau) e GTumor+5-FU+Prob (40% de adenoma tubular de baixo grau e 60% de carcinoma in situ). Concluimos que a suplementação com probiótico tem potencial para diminuir a formação de criptas aberrantes e amenizar a progressão da malignidade do tumor, potencializando o efeito antitumoral da quimioterapia com 5-Fluorouracil nos segmentos cólicos.

Made available in DSpace on 2017-07-10T18:13:29Z (GMT). No. of bitstreams: 1 Milton Cezar de Moura.pdf: 472197 bytes, checksum: ab2bbd7e5f98658de30016d4f0740ac7 (MD5) Previous issue date: 2011-08-11 / This study aimed to evaluated the use of probiotics Bacillus cereus var. Toyo and Bacillus subtilis C-3102 added to the diet for juvenile Nile tilapia strain of GIFT (Genetically Improved Farmed Tilapia) in order to verify the colonization of the gut epithelium and water, the influence on the bacterial microflora, the production performance, the indexes of body composition, the chemical composition and parameters of water quality. Two experiments were conducted: culture in cages with young individuals by 127 days (E1) and culture in ponds with adults by 201 days (E2). There was used the Center for Environmental Research in Aquaculture (CPAA/Toledo-PR) twenty ponds with 8.4 m3 of water and cages with useful volumes of 0.175 m3. The fishes from each experiment were randomly assigned to four treatments with five replicates. The treatments consisted of diets for each phase of commercial cultivation, added 0.5% B. cereus var. Toyoi (BC), 0.5% B. subtilis C-3102 (BS), 0.5% of the combination of two probiotics (BC+BS) and without addition probiotics (SP). In experiment E1, it was found that B. cereus var. Toyo and B. subtilis C-3102 colonized the gut epithelium of fishes and water culture, but did not influence the other parameters (P > 0.05). Survival ranged from 80% to 90% and feed conversion of 1.69 ± 0.29 to 1.96 ± 0.94 for treatments BS and SP. The CP levels ranged from 13.08% to 13.78% for BC and BC+BS and vicerosomatic index ranged from 9.71% to 10.9% for BS and BS+BC, respectively. In experiment E2 was also observed that the probiotics colonize the intestines and water cultivation, but did not influence the other parameters (p > 0.05). In this experiment, the indexes of production performance were influenced by the average temperature of the water that stood at 20.7ºC and is below the optimal range for the species. Weight and specific growth of the fishes ranged from 324.81 g day-1 and 398.51 g day-1, and 0.62% day-1 to 0.73% day-1, respectively, for BC and BS. Feed conversion ranged from 2.40 ± 0.24 to 2.92 ± 0.41 for BS and BC and survival ranged from 84.5% to 86% for BC and SP. The probiotics B. cereus var. Toyo and B. subtilis C-3102 used individually and in combination colonized the gut epithelium of fishes and cultivation water, however, did not influence the gut microflora, the production performance, the indexes of body composition, the chemical composition and parameters of water quality. / O presente objetivou avaliar a utilização dos probióticos Bacillus cereus var. Toyoi e Bacillus subtilis C-3102 adicionados à dieta para juvenis de tilápia do Nilo da linhagem GIFT (Genetically Improved Farmed Tilapia), a fim de verificar a colonização do epitélio intestinal e água do cultivo, a influência sobre a microflora bacteriana, o desempenho zootécnico, os índices de composição corporal, composição centesimal e os parâmetros da qualidade da água. Realizaram-se dois experimentos: cultivo em tanques rede com indivíduos jovens por 127 dias (E1) e cultivo em viveiros com indivíduos adultos por 201 dias (E2). Utilizou-se no Centro de Pesquisa em Aquicultura Ambiental (CPAA/Toledo-PR), vinte viveiros escavados com 8,4 m3 de água e tanques rede com volumes úteis de 0,175 m3. Os peixes de cada experimento foram distribuídos aleatoriamente em quatro tratamentos com cinco repetições. Os tratamentos foram constituídos de rações comerciais para cada fase de cultivo, adicionadas de 0,5% de B. cereus var. Toyoi (BC), 0,5% de B. subtilis C-3102 (BS), 0,5% da combinação dos dois probióticos (BC+BS) e sem adição de probióticos (SP). No experimento E1, verificou-se que os probióticos contendo B. cereus var. Toyoi e B. subtilis C-3102 colonizaram o epitélio intestinal dos peixes e a água do cultivo, mas não influenciaram os demais parâmetros analisados (p > 0,05). A sobrevivência variou de 80% a 90% e a conversão alimentar de 1,69 ± 0,29 a 1,96 ± 0,94 para os tratamentos BS e SP. Os níveis de PB variaram de 13,08% a 13,78% para BC+BS e BC e o índice vicerosomático variou de 9,71% a 10,9% para BC+BS e BS, respectivamente. No experimento E2, observou-se também, que os probióticos colonizaram o intestino e a água do cultivo, mas não influenciaram os demais parâmetros (p > 0,05). A temperatura média da água ficou em 20,7ºC, estando abaixo da faixa ótima para a espécie. O peso e o crescimento específico dos peixes apresentaram variação de 324,81 g dia-1 e 398,51 g dia-1; e 0,62 % dia-1 a 0,73 % dia-1, respectivamente para BC e BS. A conversão alimentar variou de 2,40 ± 0,24 a 2,92 ± 0,41 para BS e BC e a sobrevivência variou de 84,5% a 86% para BC e SP. Os probióticos B. cereus var. Toyoi e B. subtilis C-3102 utilizados individualmente e combinados realizaram a colonização do epitélio intestinal dos peixes e da água do cultivo, contudo, não influenciaram a microflora bacteriana intestinal, o desempenho zootécnico, os índices de composição corporal e centesimal e os parâmetros de qualidade da água.

The aim of the study was to evaluate the antimicrobial activity of the etanolic extract of the Annona crassiflora Mart., against the microrganisms of the endodontic microflora, aerobic, Enterococcus faecalis, Pseudomonas aeruginosa, fungi, Candida albicans and anaerobes, Prevotela melanogenicus, Clostridium sp., and Bacteroides fragilis. The methodology used involved the preparation of the etanolic extract, the liquid-liquid parttion and cromatographic purification using G 60 silica gel, silica gel with 10% of KOH and activated silica gel and LH-20 Sephadex. Procedure of purification was followed by antimicrobial activity bioassay. The identification of chemical compounds from the active fractions of the A. crassiflora were made using the espectroscopic method of magnetic resonance of 1H and 13C. The antimicrobial assay activity, for the aerobic microrganisms, E. faecalis, P. aeruginosa and fungi, C. albicans the diffusion method in disc was used. For the anaerobic microrganisms P. melanogenicus, Costridium sp., and B. fragillis the the diffusion method in broth tioglicolato was used. Also the minimun inibitory concentration (MIC) of the active fractions from the wood, bark and stem extracts of the A. crassiflora were determined and compare with the minimum inibitory concentratioon (MIC) of the standards drugs (Fluconazol, Ofloxacin, Metronidazole) and Calcium Hidroxyde used in this study. The antimicrobial evaluation tests demonstrated that the fractions of bark and wood ethanolic extracts the A. crassiflora Mart., did not present antimicrobial activity against the E. faecalis, however, presented positive activity but presented antimicrobial activity against the P. aeruginosa and the C. albicans. This study also demonstrated that the bark and stem fractions of the ethanolic extracts of the Annona crassiflora Mart., presented antimicrobial activity against the P melanogenicus, Clostridium sp., and B. fragilis, however bigger expectrum antimicrobial activity was for the Clostridium sp. Of the active fractions of the extract of the A.crassiflora Mart., A6, A10, A11 and A13 were gotten the acetogenin Goniodonin that presented in the samples A11 and A13 a CIM=1000μg/mL for C. albicans and Clostridium sp, CIM=2000μg/mL for P. aeruginosa and B. fragilis and CIM>2000μg/mL for P. melanogeniccus while in the samples A6 and A10 μg/mL for C presented a CIM=2000. albicans, P. aeruginosa and B. fragilis, CIM>2000μg/mL para P. melanogenica and CIM=62μg/mL. The samples A3, A7 and A12 had presented one expectro of similar RMN of the acetogenins without the presence of the lactônico ring and had demonstrated the same antimicrobiana activity for the P.aeruginosa, C. albicans, B. fragilis (MIC=2000μg/mL) and P. melanogeniccus (MIC>2000 μg/mL) while for the Clostidium sp the samples A3 and A7 had presented a MIC=62 μg/mL while a sample A12 had presented a MIC=500 μg/mL). / O objetivo deste estudo foi para avaliar a atividade antimicrobiana do extrato etanólico da Annona crassiflora Mart., frente a microrganismos da microflora endodôntica. Foram utilizados os microrganismos aeróbios, Enterococcus faecalis, Pseudomonas aeruginosa, fungo Candida albicans e anaeróbios, Prevotella melanogenica, Clostridium sp., e Bacteróides fragilis. A metodologia utilizada envolveu a preparação do extrato etanólico, a partição líquido-líquido e separação por meio de cromatografia em colunas usando como suporte o gel de sílica G 60, gel de sílica G-60 com 10% de KOH e Sephadex LH-20. O procedimento de purificação foi acompanhado pelo bioensaio da atividade antimicrobiana. A identificação dos compostos químicos das frações ativas do extrato da madeira e da casca da raiz da A.crassiflora foi feita pelo uso das técnicas espesctroscópicas de ressonância magnética nuclear de 1H e 13C, IV e UV e espectrometria de massa. No ensaio de atividade antimicrobiana, para os microrganismos aeróbios, P. aeruginosa e E. faecalis e fungo C.albicans, foi adotado o método de difusão em meio sólido pela técnica do disco. Para os microrganismos anaeróbios P. melanogenica, Clostridium sp e B. fragilis foi utilizado o método da difusão em caldo tioglicolato. Também foi determinada a concentração inibitória mínima (CIM) das frações ativas do extrato da madeira e da casca da raiz da Annona crassiflora comparando-se com a concentração inibitória mínima (CIM) das drogas padrões (Fluconazol, Ofloxacina, Metronidazol) e com o Hidróxido de Cálcio utilizadas neste estudo. Os testes de avaliação antimicrobiana demonstraram que as frações do extrato etanólico da casca da raiz e madeira da raiz da A. crassiflora Mart. não apresentaram nenhuma ação antimicrobiana frente ao E. faecalis, entretanto, apresentaram resultado positivo quanto à ação antimicrobiana frente a P. aeruginosa e a C. albicans. Este estudo também demonstrou que as frações do extrato etanólico da casca da raiz e madeira da raiz da A. crassiflora Mart. apresentaram ação antimicrobiana frente à P. melanogenica, Clostridium sp, e B. fragilis, demonstrando ter no entanto maior espectro antimicrobiano para o Clostridium sp. Das frações ativas do extrato da A. crassiflora Mart., A6, A10, A11 e A13 foi obtida a acetogenina Goniodonina que apresentou nas amostras A11 e A13 um CIM=1000μg/mL para C. albicans e Clostridium sp, CIM=2000μg/mL para P. aeruginosa e B. fragilis e CIM>2000μg/mL para P. melanogenica enquanto nas amostras A6 e A10 apresentou um CIM=2000 μg/mL para C. albicans, P. aeruginosa e B. fragilis, CIM>2000μg/mL para P. melanogenica e CIM=62μg/mL. As amostras A3, A7 e A12 apresentaram um espectro de RMN semelhante das acetogeninas sem a presença do anel lactônico e demonstraram a mesma atividade antimicrobiana para a P.aeruginosa, C. albicans, B. fragilis (CIM=2000μg/mL) e para a P. melanogenica (CIM>2000 μg/mL). Para o Clostidium sp as amostras A3 e A7 apresentaram CIM=62 μg/mL enquanto a amostra A12 apresentou um CIM =500μg/mL.