Obligate intracellular pathogens must invade host cells to survive and pose a global health risk. As such, internalization is a critical life stage and represents an excellent therapeutic target. Oxidoreductase exploitation is a thematic invasion strategy among obligate intracellular pathogens. Delineating the mechanisms and proteins mediating this exploitation could identify novel therapeutic targets for many important pathogens. Anaplasma phagocytophilum infects neutrophils by an incompletely defined mechanism, resulting in the emerging potentially fatal disease, human granulocytic anaplasmosis. The bacterial adhesin, Asp14, contributes to invasion by virtue of its C-terminus engaging an unknown receptor. Yeast two-hybrid analysis identified protein disulfide isomerase (PDI) as a putative Asp14 binding partner. Co-immunoprecipitation confirmed this interaction and identified the Asp14 C-terminus as critical to it. PDI reductase activity inhibition impaired bacterial infection of, but not binding to, host cells. A. phagocytophilum failed to productively infect myeloid-specific PDI conditional knock-out mice. This is the first demonstration of microbial PDI exploitation in vivo. Infection of PDI inhibited cells was rescued when bacterial, but not host surfaces were reduced with the reducing agent tris(2-carboxyethyl)phosphine (TCEP). Furthermore, TCEP restored bacterial infectivity after Asp14 inhibition using an antibody that reduces infection. Mutational analyses identified Asp14 residues critical for binding PDI. These data demonstrate that Asp14 binds and brings PDI to disulfide bonds within A. phagocytophilum surface protein(s) that it reduces, enabling infection. Targeting the Asp14 C-terminus could benefit approaches to prevent/treat granulocytic anaplasmosis. A similar approach would identify proteins from other obligate intracellular pathogens that could prove to be protective targets.

Thauera aminoaromatica MZ1T is an exopolysaccharide (EPS)-producing Gram negative bacterium isolated from the wastewater treatment plant of a major industrial chemical manufacturer as the causal agent for poor sludge dewatering. It shares common features with other known Thauera spp. (i.e. Thauera aromatica, and Thauera selenatis), being capable of degrading aromatic compounds anaerobically and using acetate and succinate as carbon sources. It is unique among the Thauera spp. in its production of abundant EPS which results in viscous bulking and poor sludge dewaterability. In this respect, it is similar to Azoarcus sp. EbN1 and BH72. Thaueran is the proposed name for EPS produced by MZ1T for research purpose. The focus of this research is to fully characterize the microorganism and identify and characterize the genes responsible for thaueran synthesis and export through bioinformatics, transposon mutagenesis, gene clone and expression, reverse transcriptase quantitative real time PCR, and genome sequencing and annotation. Ultimately, this knowledge will contribute to control of viscous bulking and sludge dewatering problems. However, a broader range of important environmental biotechnical processes may be forthcoming from understanding thaueran synthesis. They may include thaueran remedial solutions for heavy metal and radionuclide immobilization, anaerobic carbon channeling and sequestration, greenhouse gas mitigation through acetate incorporation into thaueran, and novel applications such as thaueran-mediated wound healing. Sequencing of MZ1T genome has been accomplished through collaboration with the Joint Genome Institute (JGI). The genome size is 4.5 Mbp, GC content is 68.3%, and there are 4,092 protein coding genes. Three putative thaueran gene clusters were found within the genome. One tight cluster with a size of 20.67kb encoding 14 genes may contain most necessary genes for thaueran formation and export. Another two clusters are loosely organized. Through transposon mutagenesis, mutants not producing abundant thaueran and not flocculating have been obtained and verified, and were further used in reverse transcriptase quantitative real time PCR to compare the differential expression levels of the presumable EPS genes among mutants, wild type MZ1T and under different growth conditions. The results indicated a correlation of the expression level of the test genes and the abundance of EPS.

Biofilms are communities of sessile microorganisms attached to a surface and imbeddedin a matrix of extracellular polysaccharide substances. These communities can be foundin diverse aquatic environments, such as in industrial pipes and in humans. By formingmicrocolony structures, which are highly resistant to adverse physical conditions as wellas antimicrobial agents, biofilms are very problematic when associated with e.g.persistent infections. In order to find new ways of controlling biofilm growth, theprocesses involved in biofilm development must be investigated further. The maininterest of this study is the occurrence of void formation inside biofilms. Thisphenomenon has been observed in several studies and has been correlated to cell deathinside the microcolonies. The occurrence of cell death has recently been associated withthe presence of nitric oxide in the biofilm. In this study, the implications of nitric oxideaccumulation on biofilm development were investigated using an individual-basedmodel. Specifically, the role of nitric oxide in void formation was considered. A largenumber of simulations were run using different parameter settings in order to determine ifnitric oxide could account for the occurrence of void formation observed experimentally.The general predictions made by the model system showed agreement to someexperimental data, but not to others. Sloughing, the detachment of chunks of cells fromthe biofilm, was observed in the majority of simulations. In some cases, the model alsopredicted the presence of live cells inside the voids, which has been observedexperimentally.

No description available.

<p>Biofilms are communities of sessile microorganisms attached to a surface and imbeddedin a matrix of extracellular polysaccharide substances. These communities can be foundin diverse aquatic environments, such as in industrial pipes and in humans. By formingmicrocolony structures, which are highly resistant to adverse physical conditions as wellas antimicrobial agents, biofilms are very problematic when associated with e.g.persistent infections. In order to find new ways of controlling biofilm growth, theprocesses involved in biofilm development must be investigated further. The maininterest of this study is the occurrence of void formation inside biofilms. Thisphenomenon has been observed in several studies and has been correlated to cell deathinside the microcolonies. The occurrence of cell death has recently been associated withthe presence of nitric oxide in the biofilm. In this study, the implications of nitric oxideaccumulation on biofilm development were investigated using an individual-basedmodel. Specifically, the role of nitric oxide in void formation was considered. A largenumber of simulations were run using different parameter settings in order to determine ifnitric oxide could account for the occurrence of void formation observed experimentally.The general predictions made by the model system showed agreement to someexperimental data, but not to others. Sloughing, the detachment of chunks of cells fromthe biofilm, was observed in the majority of simulations. In some cases, the model alsopredicted the presence of live cells inside the voids, which has been observedexperimentally.</p>

Xylella fastidiosa é uma bactéria gram-negativa que coloniza os vasos de xilema das plantas, podendo ser patogênica para algumas culturas de importância econômica. Esta bactéria é transmitida por cigarrinhas (Hemiptera: Auchenorrhyncha) da superfamília Cercopoidea (Aphrophoridae, Cercopidae e Clastopteridae) e família Cicadellidae (subfamília Cicadellinae). Na Itália este fitopatógeno esta causando uma nova doença em plantas de oliveira, denominada síndrome do declínio rápido da oliveira. Recentemente também foi detectada a ocorrência desta bactéria associada a sintomas de requeima foliar na Argentina e no Brasil. Sendo assim, surgiu a necessidade de realizar levantamentos de espécies de cigarrinhas que ocorrem nas regiões onde estão localizados os pomares de oliveira na Serra da Mantiqueira, nos estados de São Paulo e Minas Gerais, na faixa de 800 a 1800 m de altitude. O objetivo deste trabalho foi realizar a análise faunística das espécies de cigarrinhas, como também mensurar a similaridade da comunidade de espécies encontradas entre os pomares, e indicar possíveis vetores de Xylella fastidiosa para a cultura da oliveira. Foram selecionadas sete pomares de oliveira localizados na Serra da Mantiqueira, seguindo um gradiente altitudinal, sendo estes: Wenceslau Braz (MG-1780 m), São Bento do Sapucaí (Entre Vilas) (SP-1602 m), São Bento do Sapucaí (São José) (SP-1512 m), Maria da Fé (MG), com três pomares (área do Lago-1329 m, área da Suíça-1318 m, área da Atemoia-1310 m) e Cabreúva (SP-883 m). Para a captura de cigarrinhas, foram instalados em cada pomar nove cartões adesivos amarelos, em duas alturas, 0,8 m e 1,6 m do nível do solo, totalizando 18 cartões por pomar, que foram trocados a cada 15 dias no período de junho de 2015 a maio de 2017. Coletaram-se 40.324 indivíduos de 270 espécies incluídos em Auchenorrhyncha, distribuídos em 12 famílias, com maior frequência de indivíduos do grupo Cicadellidae (68%), e Cicadellinae (43% de todos os cicadelídeos coletados). A análise faunística indicou 44 espécies predominantes, sendo 20 pertencentes a grupos de potenciais vetores. Dentre elas, Clastoptera sp. 1 (Clastopteridae), Macugonalia cavifrons e Scopogonalia sp. 1 (Cicadellinae) foram predominantes em todos os pomares, exceto no pomar de Cabreúva, sendo que a primeira coloniza a oliveira. Os maiores índices de diversidade, riqueza e equitabiliade foram obtidos para os pomares de Wenceslau Braz, São Bento do Sapucaí (São José) e São Bento do Sapucaí (Entre Vilas) (0,87), respectivamente. A maior similaridade de Jaccard foi encontrada entre os pomares de Maria da Fé (55%). A maior similaridade de Morisita foi encontrada entre os pomares de São Bento do Sapucaí (São José) e Maria da Fé (área da Suíça) (91%) e a menor entre os pomares de Cabreúva e São Bento do Sapucaí (Entre Vilas) (6%). Para algumas espécies predominantes de Cicadellinae e Cercopoidea, houve diferença na eficiência de captura por armadilhas adesivas posicionadas em alturas distintas na copa das oliveiras. / Xylella fastidiosa is a gram-negative bacterium that colonizes the xylem vessels of plants and may be pathogenic to some crops. This bacterium is transmitted by leafhoppers (Hemiptera: Auchenorrhyncha) from the superfamily Cercopoidea (Aphrophoridae, Cercopidae and Clastopteridae) and family Cicadellidae (subfamily Cicadellinae). In Italy this phytopathogen is causing a new disease in olive trees, named \'olive quick decline syndrome\'. Recently, the occurrence of this bacterium associated to a \'leaf scorch\' was also detected in Argentina and Brazil. Thus, it is necessary to carry out surveys of Auchenorrhyncha species in olive orchards in the region of the Mantiqueira Mountain Range, where the olives are grown in the states of São Paulo and Minas Gerais, at 800 to 1800 m altitude range. The objectives of this work were to do faunistic analysis of Auchenorrhyncha species, to measure species similarity of the community of Auchenorrhyncha among orchards, and to indicate possible vectors of Xylella fastidiosa in olives. Seven olive orchards located in the Mantiqueira Mountain Range were selected, following an altitudinal gradient: Wenceslau Braz (MG-1780 m), São Bento do Sapuccaí (EntreVilas) (SP-1602 m), São Bento do Sapucaí (São José) (SP-1512 m), Maria da Fé (MG), with three orchards (area of Lago-1329 m, area of Suíça-1318 m, area of Atemoia-1310 m) e Cabreúva (SP-883 m). Nine yellow stick traps were installed in each orchard at two heights, 0.8 m and 1.6 m above soil level, totaling 18 cards per orchard, which were replaced fortnightly from June/2015 to May/2017. A total of 40,324 individuals from 270 species included in Auchenorrhyncha were collected, distributed in 12 families, with the highest frequency of individuals in the group Cicadellidae (68%) and group Cicadellinae (43% of all Cicadellidae). The faunistic analyses classified 44 species as predominant, 20 belonging to groups of possible vectors of X. fastidiosa. Among them, the species Clastoptera sp. 1 (family Clastopteridae), Macugonalia cavifrons and Scopogonalia sp. 1 (subfamily Cicadellinae) are predominant in all orchards, except in the Cabreúva orchard. Nymphs of Clastoptera sp. 1 were observed developing on young olive branches. Higher diversity, richness and equitability indices were observed in Wenceslau Braz, São Bento do Sapucaí (São José) and São Bento do Sapucaí (Entre Vilas) orchards, respectively. Jaccard\'s greatest similarity was found between Maria da Fé orchards (55%). The greatest similarity of Morisita was found between the orchards of São Bento do Sapucaí (São José) and Maria da Fé (91%) and the lowest one between orchards of Cabreúva and São Bento do Sapucaí (Entre Vilas) (6%). For some predominant species of Cicadellinae and Cercopoidea, capture efficiency was influenced by trap height on the olive tree canopy.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-07T13:54:35Z No. of bitstreams: 1 Fernanda Silva de Meirelles.pdf: 599414 bytes, checksum: be09ab548e388263d8d38ab7cabb0cf5 (MD5) / Made available in DSpace on 2016-10-07T13:54:35Z (GMT). No. of bitstreams: 1 Fernanda Silva de Meirelles.pdf: 599414 bytes, checksum: be09ab548e388263d8d38ab7cabb0cf5 (MD5) Previous issue date: 2010-02-26 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / In Brazil, it is considered that the annual production of tilapias overcomes 100 thousand tons, mainly due to the productive potential of the Northeast area that answered for 41% of the total production of 70.000 tons in 2004, generating income of US$ 200.000.000,00, due to your climatic conditions, technology readiness and consumption market, regional and national in expansion. The intensification of cultivations has as consequence the increase of organic matter, that it favors the multiplication of microorganisms, making possible him/it supplies of diseases in the occurrence of adverse situations to the fish. Tilapia (Oerochromis niloticus) of cultivations of the state of Pernambuco they were appraised with objective of determining the frequency of bacterial diseases. For so much, samples of several farms were analyzed, in the several aspects, with relevance for the bacterial agents. The tilapia were examined with relationship to the streptococcus presence, víbrios, coliform, pseudomonas and aeromonas. Clinical exams and autopsy were accomplished, for microscopic analysis of the lesions and better definition of the probable agent etiological. The analyses were accomplished at the Laboratory of Sanity of Aquatic Animals (LASAq) of the Rural Federal University of Pernambuco (UFRPE) and in the Center of Development and Diffusion of Technology in Aquaculture of the State University of Bahia (UNEB). The principal pathological agents were víbrios and aeromonas, that are pathogens responsible opportunists for significant losses, being identified the following species in 46 isolated: V. natriegens (1/46), V. metschnikovii (2/46), V. halioticoli (1/46), V. fischeri (2/46), V. mimicus (23/46), V. diabolicus (1/46), V. furnissi (1/46), V. cholerae o1 (1/46), V. scophthalmi (4/46), V. proteolyticus (3/46), V. argarivorans (1/46), V. ordalii (2/46) and Vibrio spp. ( 3/46). The isolated vibrios presented larger sensibility to the antimicrobianos enrofloxacina (100%) and florfenicol (98,18%) and in decreasing order, gentamicina (90,91%), cotrimoxazol (sulfametoxazol+trimetroprim) (76,36%), tetraciclin (67,27%), eritromicin (30,91%) and amoxilina (3,64%). The isolated ones tested they came 100% resistant the penicillin. In this study, 96,4% (53/55) of the víbrios they presented index multiple resistance to antibiotics (MAR), superior to 0,22, characterizing multiple resistance. In 35 isolated the identified aeromonas were: A.caviae (1/35), A. shubertii (11/35), A. media (4/35), A. popoffii (1/35), A. sobria (3/35), A. encheleia (4/35), A. veronii (4/35) e A. jandaei (4/35) Forty were tested isolated of aeromonas with relationship to the sensibility to 08 antimicrobial, of these they showed larger sensibility the florfenicol (100,0%), enrofloxacina(95,0%), gentamicina (95,0%) and in decreasing order, cotrimoxazol (sulfametoxazol+trimetroprima) (67,5%), tetraciclina (65,0%). There was smaller sensibility the eritromicina (20,0%), penicillin (5,0%) and the amoxilina (2,5%), confirming the resistance existence. The index of multiple resistance to antibiotics (MAR) for the isolated of aeromonas it varied in an interval from 0,12 to 0,62, of these 95% (38/40) they came superior to 0,22 characterizing multiple resistance. Most of the species of isolated víbrios is not considered pathogenic for the fish (environmental), but nevertheless they can represent risk for the health of the tilapia for subject of opportunism and of the consumer, mainly if consumed raw. The isolated and identified movable aeromonas in this study are commonly considered as environmental, however many of the analyzed fish presented compatible symptomatology with the illness caused by these agents opportunists. / No Brasil, estima-se que a produção anual de tilápias supere 100 mil toneladas, principalmente devido ao potencial produtivo da região Nordeste que em 2004 respondeu por 41 % da produção total de 70.000 toneladas, gerando renda de US$ 200.000.000,00, decorrente de suas condições climáticas, disponibilidade de tecnologia e mercado de consumo, regional e nacional em expansão. A intensificação de cultivos tem como conseqüência o aumento de matéria orgânica, que favorece a multiplicação de microrganismos, possibilitando o surto de doenças na ocorrência de situações adversas aos peixes. Tilápias (Oerochromis niloticus) de cultivos do estado de Pernambuco foram avaliadas com objetivo de determinar a freqüência de doenças bacterianas. Para tanto, analisaram-se amostras de diversas fazendas, nos diversos aspectos, com relevância para os agentes bacterianos. As tilápias foram examinadas quanto à presença de estreptococos, víbrios, coliformes, pseudomonas e aeromonas. Foram realizados exames clínicos e necropsias, para análise microscópica das lesões e melhor definição do provável agente etiológico. As análises foram realizadas no Laboratório de Sanidade de Animais Aquáticos (LASAq) da Universidade Federal Rural de Pernambuco (UFRPE) e no Centro de Desenvolvimento e Difusão de Tecnologia em Aqüicultura da Universidade Estadual da Bahia (UNEB). Os principais agentes patológicos foram víbrios e aeromonas, que são patógenos oportunistas responsáveis por perdas significantes, sendo identificadas as seguintes espécies em 46 isolados: Vibrio natriegens (1/46), V. metschnikovii (2/46), V. halioticoli (1/46), V. fischeri (2/46), V. mimicus (23/46), V. diabolicus (1/46), V. furnissi (1/46), V. cholerae O1 (1/46), V. scophthalmi (4/46), V. proteolyticus (3/46), V. argarivorans (1/46), V. ordalii (2/46) e Vibrio spp.(3/46). Os vibrios isolados apresentaram maior sensibilidade aos antimicrobianos enrofloxacina (100%) e florfenicol (98,18%) e em ordem decrescente, gentamicina (90,91%), cotrimoxazol (sulfametoxazol+trimetroprima) (76,36%), tetraciclina (67,27%), eritromicina (30,91%) e amoxilina (3,64%). Os isolados testados apresentaram-se 100% resistentes a penicilina. Neste estudo, 96,4% (53/55) dos víbrios apresentaram índice múltipla resistência a antibióticos (MAR), superior a 0,22, caracterizando múltipla resistência. Em 35 isolados as aeromonas identificadas foram: Aeromonas caviae (1/35), A. shubertii (11/35), A. media (4/35), A. popoffii (1/35), A. sobria (3/35), A. encheleia (4/35), A. veronii (4/35) e A. jandaei (4/35). Foram testados 40 isolados de aeromonas quanto à sensibilidade a oito antimicrobianos, tendo mostrado maior sensibilidade a florfenicol (100,0%), enrofloxacina(95,0%), gentamicina (95,0%), cotrimoxazol (sulfametoxazol+trimetroprima) (67,5%), tetraciclina (65,0%). Houve menor sensibilidade a eritromicina (20,0%), penicilina (5,0%) e a amoxilina (2,5%), confirmando a existência de resistência. O índice MAR para os isolados de aeromonas variou de 0,12 a 0,62, dos quais 95% (38/40) apresentaram índices superiores a 0,22, caracterizando multiresistência. A maioria das espécies de víbrios isoladas não é considerada patogênica para os peixes (ambientais), mas ainda assim podem representar risco para a saúde das tilápias por questão de oportunismo e do consumidor, principalmente se consumido cru. As aeromonas móveis isoladas e identificadas neste estudo são comumente consideradas como ambientais, porém muitos dos peixes analisados apresentaram sintomatologia compatível com a enfermidade causada por estes agentes oportunistas.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-14T12:43:53Z No. of bitstreams: 1 Josenaldo Silva Macedo.pdf: 689898 bytes, checksum: 3215e2e1e0646d554d6f16686dd6361d (MD5) / Made available in DSpace on 2016-10-14T12:43:53Z (GMT). No. of bitstreams: 1 Josenaldo Silva Macedo.pdf: 689898 bytes, checksum: 3215e2e1e0646d554d6f16686dd6361d (MD5) Previous issue date: 2011-02-24 / The present work was designed to investigate the incidence of the bacterium Helicobacter spp. In cats from the Recife city, Pernambuco State. The stomach samples were collected from 119 cats, without distinction between race or age, being 56 males and 63 females originated from the Environmental Survey Center (CVA), captured in the city of Recife.. The urease quick test and the histopathological examination using both the Hematoxilin-Eosin (HE) and Giemsa stains were performed. The urease quick test showed that 83,92% of the male (47/56) and 80,95% of the female (51/63) samples were positive for Helicobacter, with a total of 82,35% (98/119) of the population being positive. The Giemsa analysis demonstrated that in 73,21% of the males (41/56) and 74,60% of the females (47/63) samples the Helicobacter spp. Bacteria was found, and the total positive sample in the tested population was 73,94% (88/119). During the analysis performed using the HE stain, was observed that 59,66% (71/119) of the samples showed histopathological alterations, such as: lymphoplasmocitic infiltration with a lymphoid foliculum, microabscess, necrosis, hypotrophy, and in 40,34% of the samples(48/119) no alterations were observed. Since the Helicobacter spp. Microorganisms are able to cause gastric inflammation in cats, having showed , the research studies concerning the clinical importance and elucidate the gastric neoplasic genesis in these animals. It coud conclude that cats originated from Recife city, Pernambuco state, shows high incidence of helicobacteriosis. / O presente trabalho foi delineado para investigar a incidência da bactéria Helicobacter spp. em gatos oriundos da cidade do Recife do estado de Pernambuco. As amostras de estômagos analisadas, foram coletados de 119 gatos sem raça definida e idade, sendo 56 machos e 63 fêmeas provenientes do Centro de Vigilância Ambiental (CVA), capturados na cidade do Recife. Foram realizados para pesquisa de Helicobacter spp., os métodos de teste rápido da urease, o exame histopatológico corado pela técnica hematoxilina-eosina (HE) e pelo método de coloração Giemsa. As análises pelo teste rápido da urease mostraram que em 83,92% das amostras dos machos (47/56) e em 80,95% das amostras das fêmeas (51/63), foi detectada reação positiva para Helicobacter, sendo 82,35% (98/119) na totalidade das amostras. As análises pelo método de coloração pelo Giemsa mostraram que em 73,21% das amostras dos machos (41/56) e em 74,60% das amostras das fêmeas (47/63), foi encontrada a bactéria Helicobacter spp. , sendo 73,94% (88/119) na totalidade das amostras. Na análise das amostras coradas pela técnica de coloração hematoxilia-eosina (HE), verificou-se que em 59,66% das amostras (71/119) apresentaram alterações histopatológicas como: infiltrado linfoplasmocitário com folículo linfóide, microabcesso, necrose, hipotrofia; e em 40,34% das amostras (48/119) não apresentaram alterações histológicas. Dessa forma, os microorganismos do gênero Helicobacter spp. são capazes de gerar inflamação gástrica no gato, sendo necessário cada vez mais aprofundar as pesquisas a respeito da importância clínica e da gênese de neoplasias gástricas nesses animais. Concluiu-se neste trabalho que gatos oriundos da cidade do Recife, estado de Pernambuco apresentam alta incidência de helicobacteriose.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-20T14:50:29Z No. of bitstreams: 1 Monique Monteiro Pinto.pdf: 848527 bytes, checksum: 1b2bc1d9050d7a02d03c3c3faf35a485 (MD5) / Made available in DSpace on 2016-10-20T14:50:29Z (GMT). No. of bitstreams: 1 Monique Monteiro Pinto.pdf: 848527 bytes, checksum: 1b2bc1d9050d7a02d03c3c3faf35a485 (MD5) Previous issue date: 2013-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Probiotics are responsible for a new idea of using preventive in aquaculture, aiming at sustainable farming, environmental balance and of its products generated. However, there are few knowledge on the subject, despite its widespread use in shrimp farming, randomly in the most of times. Because of large demand from the sector for new alternatives, to maintain animal health, reduce the chance of having disease problems that the study aimed to identify and test bacteria with potential probiotic isolated from the intestinal tract of cultured shrimp (Litopenaeus vannamei) in salt water coming from commercial farm in the state of Pernambuco. Has been collected 90 animals, 50 in rainy season and 40 in dry season. The shrimps collected during the rainy season had weight of 9.50 ± 0.52 g and length of 6.72 ± 0.84 cm, while the animals from the dry period had weight of 9.76 ± 0.24 g and a length of 6, 54 ± 0.42 cm. Were obtained 72 bacterial isolates then performed the Gram 31 were tested in vitro against dual-known pathogenic species, Vibrio parahaemolyticus (ATCC 17802) and Vibrio vulnificus (ATCC 27562). Eight isolates (28.81%) showed antibacterial activity to at least one pathogen. There was no significant difference (P <0.05) in the proportion of isolates potential probiotics obtained in different seasons, rainy and dry. Isolates that showed the best results in the antagonism test in vitro were identified as Lactobacillus paracasei, Bacillus cereus, Lactococcus lactis subsp. lactis, Lactobacillus crispatus and Lactobacillus brevis. The largest zone of inhibition observed in the antagonism test was produced by Lactobacillus crispatus opposite V. vulnificus. All bacteria tested against the pathogens studied produced inhibition zone, however it is suggested the testing of antagonism in vivo to be proven the effectiveness of probiotic bacteria for marine shrimp. / Os probióticos são responsáveis por uma nova ideia de utilização de preventivos na aquicultura, visando a sustentabilidade do cultivo, equilíbrio do meio ambiente e dos seus produtos gerados. Entretanto, ainda são poucos os conhecimentos a respeito do tema, apesar da sua vasta utilização na carcinicultura, muitas vezes de forma aleatória. Em virtude da grande demanda do setor por novas alternativas para manter a saúde animal, que se objetivou identificar e testar bactérias, com potencial probiótico, isoladas do trato intestinal de camarões cultivados (Litopenaeus vannamei) em água salgada, oriundos de fazenda comercial do estado de Pernambuco.Foram coletados 90 animais, sendo 50 em período chuvoso e 40 em período de estiagem, todos apresentando-se saudáveis. Os camarões coletados no período chuvoso apresentaram peso de 9,50 ± 0,52g e comprimento de 6,72 ± 0,84cm, enquanto os animais analisados no período de estiagem o peso de 9,76 ± 0,24g e comprimento de 6,54 ± 0,42cm. Foram obtidos 72 isolados bacterianos, que foram reduzidos para 31 após a análise do Gram, os quais foram testados in vitro frente as espécies patogênicas, Vibrio parahaemolyticus (ATCC 17802) e Vibrio vulnificus (ATCC 27562). Oito isolados (28,81%) apresentaram atividade antibacteriana a pelo menos um patógeno. Não houve diferença significativa (P<0,05) na proporção de isolados potenciais probióticos obtidos nas distintas épocas do ano, chuvosa e seca. Os isolados que apresentaram os melhores resultados no teste de antagonismo in vitro foram identificados como Lactobacillus paracasei, Bacillus cereus, Lactococcus lactis subsp. lactis, Lactobacillus crispatus e Lactobacillus brevis. O maior halo de inibição observado no teste de antagonismo foi produzido por Lactobacillus crispatus frente ao V. vulnificus. Todas as bactérias testadas contra os patógenos estudados produziram halo de inibição, contudo sugere-se a realização de testes de antagonismo in vivo para que seja comprovada a efetividade das bactérias como probióticas para camarão marinho

Orientador: Adriana Zerlotti Mercadante / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-16T15:01:11Z (GMT). No. of bitstreams: 1 Mandelli_Fernanda_M.pdf: 677502 bytes, checksum: 40f71e5be548712e1d9c40d24476ba1b (MD5) Previous issue date: 2010 / Resumo: Os carotenoides pertencem a um grupo de pigmentos naturais amplamente distribuídos na natureza, com grande diversidade de estruturas e funções. O interesse pelos carotenoides tem aumentado consideravelmente nos últimos anos pela evidência dos seus já conhecidos benefícios à saúde humana, devido às suas propriedades antioxidantes, anticarcinogênicas e imunomodulatórias. As principais fontes industriais de carotenoides são a síntese química e a extração a partir de plantas. Porém, são poucos os carotenoides que possuem métodos economicamente eficientes de produção. Uma alternativa para a obtenção natural destes pigmentos é a sua produção por micro-organismos, sendo uma das fontes as bactérias termófilas que, em sua maioria, possuem carotenoides com estrutura específica para fortalecimento da membrana celular. Assim sendo, o presente estudo tem como objetivos: identificar os carotenoides da linhagem Thermus filiformis ATCC 43280, avaliar o crescimento de massa celular, produção de carotenóides e porcentagem de proteção contra o oxigênio singlete através de diferentes condições de cultivo e verificar a atividade antioxidante do extrato rico em carotenoides na desativação do radical ABTS¿+ (2,2¿- azinobis-(3-etilbenzotiazolina-6-ácido sulfônico)) e do peróxido de hidrogênio. Para atingir os objetivos, os carotenoides foram caracterizados por cromatografia líquida de alta eficiência conectada aos detectores de arranjo de diodos e espectrômetro de massas (HPLCDAD-MS/MS), e as condições de cultivo e a interação entre as variáveis foram estudadas através de planejamento estatístico fatorial. Foram identificados nove carotenoides sintetizados pela bactéria Thermus filiformis, todos apresentando o mesmo comprimento de onda máximo a 449 ± 1 nm, que é um indicativo de um cromóforo de 11 ligações duplas conjugadas, sendo duas em anel ?. O carotenoide majoritário identificado em T. filiformis foi termozeaxantina-15 que consiste do carotenoide zeaxantina ligada a uma glicose acilada com ácido graxo ramificado de 15 carbonos ((3R,3'R)-3'-[(6-O-13-metiltetradecanoil-ß-Dglucopiranosil) oxi]-ß,ß-caroten-3-ol) o que representa 35,3 % do conteúdo total de carotenóides. Foram também identificados os carotenoides: zeaxantina (14,4 %), zeaxantina monoglicosilada (4,7 %) e termozeaxantinas com ácidos graxos ramificados, com cadeia de cadeia de 11 (2,3 %), 12 (5,2 %), 13 (25,4 %) e 14 (9,0 %) carbonos, além de derivados dos carotenoides termozeaxantina-13 (0,8 %) e termozeaxantina-15 (2,8 %). Dentre as variáveis estudadas no planejamento experimental (temperatura, pH, quantidade de triptona, extrato de levedura e elementos traços de Nitsch¿s), a temperatura foi a que mais influenciou a quantidade de biomassa, carotenoides totais e proteção proporcionada pelos carotenoides contra oxigênio singlete. Com a utilização desta ferramenta foi possível atingir valores de 658 mg/L de biomassa seca, 1517 µg/g de carotenoides totais e porcentagem de proteção conferida ao DMA (dimetilantraceno) frente a oxidação por oxigênio singlete na ordem de 85 %. O extrato rico em carotenoides de T. filfiormis apresentou capacidade de desativar o radical ABTS¿+ com valor de capacidade antioxidante equivalente a Trolox (TEAC) de 2,87, valor este superior aos relatados para outros carotenoides como ß-caroteno (1,9) e zeaxantina (1,4). / Abstract: Carotenoids are a group of natural pigments widely distributed in the nature with great diversity of structures and functions. The interest on carotenoids has considerably increased in recent years by the evidence of its known benefits to human health, due to its antioxidant, anticarcinogenic and immunomodulatory properties. The main industrial sources of carotenoids are chemical synthesis and extraction from plants. However, there are few carotenoids which production is economically feasible. An alternative to obtain these natural pigments is their production by microorganisms, and one of the sources is the thermophilic bacterium that produces carotenoids with specific structure for the strength of cell membrane. Therefore the aims of this work were to evaluate the biomass, carotenoid production and quenching of singlet oxygen by the strain Thermus filiformis ATCC 43280 through different cultivation conditions and to verify the antioxidant activity by scavenging of the ABTS¿+ (2,2'-azino-bis(3-ethylbenzthiazoline-6-Sulfonic Acid)) radical and hydroxy peroxide deactivation. In order to achieve these aims, the carotenoids were characterized by high-performance liquid chromatography connected to a photodiode array and mass spectrometry detectors (HPLC-DAD-MS/MS), and the cultivation conditions and the interactions between the variables were studied using a fractionated factorial design. Nine carotenoids synthesized by Thermus filiformis, were identified, all showing the same maximum absorbance wavelength, indicating the presence of a chromophore of 11 conjugated double bounds, being two in the ?-ring. The major carotenoid identified in T. filiformis was thermozeaxanthin-15 which is the carotenoid zeaxanthin linked to a glucose, acylated with a branched fatty acid of 15 carbons ((3R, 3'R) -3'- [(6-O-13- methyltetradecanoyl-ß-D-gluycopiranosyl) oxy]-ß,ß-carotene-3-ol) what represents 35.3 % of the total content of carotenoides. Zeaxanthin (14.4 %), zeaxanthin monoglucoside (4.7 %) and thermozeaxanthins with branched fatty acids chain with 11 (2.3 %), 12 (5.2 %), 13 (25.4 %) and 14 (90 %) carbons, besides thermozeaxanthin-13 (0,8 %) and thermozeaxanthina-15 (2,8 %) derivatives were also identified. Among the studied variables in the experimental design (temperature, pH, tryptone, yeast extract and Nitsch¿s trace elements) the temperature was the one that most influenced the biomass amount, total carotenoids and percentage of protection against singlet oxygen. Using this tool, it was possible to achieve values of 658 mg/L of dry biomass, 1517 µg/g of total carotenoids and percentage of protection of DMA (dimethylanthracene) against the oxidation by singlet oxygen about 85 %. The extract rich in carotenoids from T. filiformis showed capacity of scavenging ABTS¿+ radical with Trolox equivalent antioxidant capacity (TEAC) of 2.87, being this value higher than those reported in the literature for ß-carotene (1.9) and zeaxanthin (1.4) / Mestrado / Mestre em Ciência de Alimentos