Made available in DSpace on 2016-04-28T20:15:27Z (GMT). No. of bitstreams: 1 2007-Isabele da Costa Angelo.pdf: 1629986 bytes, checksum: 663b35706e830a57e185c3e0a68096c9 (MD5) Previous issue date: 2007-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Boophilus microplus is an ectoparasite which causes great economic losses around the world. The exclusive use of acaricides and the inadequate management have conducted the development of resistance in ticks populations, environmental and food contamination by acaricides and their residues. The use of entomopathogenic fungi to arthropods control has shown interesting responses. Several studies have proved the pathogenicity of fungi toward various ticks species. The aim of the present study was to evaluate the effect of Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus and Lecanicillium lecanii toward engorged females, eggs and larvae of B. microplus tick. The isolates were cultured on malt extract medium at 25 ? 1?C and 80% of relative humidity for 15 days. Conidial suspensions were prepared in Tween 80 water solution (0.1% v/v). There were 4 treatment groups according to the following conidial concentrations: 105, 106, 107 and 108 conidia mL-1. The control group was made up of water and Tween 80 only (0.1% v/v). Treatment was based on immersion of the specimen in 1 mL of the conidial suspension. Each treatment group was made up of 10 repetitions. Changes in biology of engorged females, eggs viability and larvae mortality, were observed every 5 days up to the 20th day after treatment. The results have shown that L. lecanii changed biological parameters in engorged females through the reduction in posture period, nutritional rate, eggs production rate, and increase of incubation period. Isaria farinosa has shown reduction in posture period and in nutritional rate. Paecilomyces lilacinus was the unique isolate able to reduce the eclosion period of larvae from infected females. Isaria fumosorosea has reduced the nutritional rate and was the unique isolate that changed significantly in eclosion period when eggs were infected. Conidial concentrations reduced reproduction capacity of engorged females. The isolates of I. fumosorosea and L. lecanii have presented major potential to control B. microplus engorged females. All tested isolates have shown pathogenicity toward unfed larvae of B. microplus after in vitro infection. / Boophilus microplus ? um ectoparasito que causa grandes perdas na pecu?ria mundial. A utiliza??o exclusiva de produtos qu?micos no controle de carrapatos, associada ao manejo inadequado tem conduzido o desenvolvimento de popula??es de carrapatos resistentes, e a contamina??o dos produtos de origem animal e do ambiente pelos seus res?duos. O uso de fungos entomopatog?nicos no controle de artr?podes tem se tornado uma abordagem cada vez mais atrativa. In?meros trabalhos comprovam experimentalmente a patogenicidade dos fungos sobre diversas esp?cies de carrapatos. O objetivo do presente trabalho foi avaliar o efeito in vitro de Isaria farinosa, I. fumosorosea, Paecilomyces lilacinus e Lecanicillium lecanii sobre f?meas ingurgitadas, ovos e larvas de B. microplus. Os fungos foram repicados em meio de cultura extrato de malte, e mantidos em c?mara climatizada sob temperatura de 25?C ? 1 e umidade relativa de 80% por quinze dias. Suspens?es conidiais foram preparadas a partir do crescimento do fungo, cujos con?dios foram adicionados a solu??o de ?gua destilada e Tween 0,1%. As concentra??es 105, 106, 107 e 108 con?dios mL-1 formaram os grupos tratamento, juntamente com o grupo controle, constitu?do por ?gua destilada est?ril e Tween 0,1%. O tratamento constituiu-se de um mililitro da concentra??o conidial testada, e cada grupo foi formado por 10 repeti??es. Os par?metros de avalia??o observados para demonstrar o efeito dos fungos sobre o carrapato foram as altera??es biol?gicas de f?meas ingurgitadas, viabilidade de ovos tratados e percentual de mortalidade de larvas, acompanhado a cada cinco dias at? o 20? dia ap?s infec??o. Os resultados mostraram que L. lecanii causou altera??es nos par?metros biol?gicos de f?meas ingurgitadas, diminuindo o per?odo de postura, o ?ndice nutricional, o ?ndice de produ??o de ovos, e aumentando o per?odo de incuba??o. Isaria farinosa mostrou redu??o no per?odo da postura e no ?ndice nutricional de f?meas ingurgitadas. P. lilacinus foi o ?nico fungo capaz de reduzir o per?odo de eclos?o das larvas provenientes da infec??o de f?meas ingurgitadas. I. fumosorosea reduziu o ?ndice nutricional e foi o ?nico isolado que causou altera??o significativa no per?odo de eclos?o de larvas provenientes da infec??o de ovos. Diferentes concentra??es dos entomomopat?genos reduziram o potencial reprodutivo das f?meas ingurgitadas dos grupos tratados. Os isolados de I. fumosorosea e L. lecanii apresentaram maior potencial de controle para f?meas ingurgitadas de B. microplus. Todos os isolados testados mostraram patogenicidade para larvas n?o alimentadas de B. microplus ap?s infec??o in vitro.

Made available in DSpace on 2016-04-28T20:16:20Z (GMT). No. of bitstreams: 1 2007-Thiago Campanharo Bahiense.pdf: 1378749 bytes, checksum: ef9fdcee9b7f5b82132b5b8d53a3fbe5 (MD5) Previous issue date: 2007-02-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Boophilus microplus can cause several cattle pathology and thus economic losses. Moreover, this tick species is able to develop resistance to acaricides. The biological control is a great promise to control cattle ticks populations. The aim of the present study was to contribute to the development of a new complementary method to control B. microplus. Therefore, it has been evaluated the compatibility between deltamethrin and the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae toward B. microplus tick in a stable test. It was used a tick strain resistant to pyrethroid and calves were artificially infested. A total of 8 calves were held in individual housing and infested with B. microplus larvae every other day for 25 days. After that, 4 treatments were realized with 2 calves per group. The control group was tested receiving just water; the second group was tested receiving M. anisopliae in a 108 conidia ml-1 suspension; the third one was tested receiving a 25 ppm deltamethrin solution; and the forth group was tested receiving a solution containing a 108 conidia ml-1 M. anisopliae suspension associated with a 25 ppm deltamethrin solution. The mortality rate was 32.57% in the second group, 38.58% in the third group, and 30.92% in the forth one. The main changes in ticks biology were a decreased nutritional rate until the forth day after treatment. The deltamethrin group was an exception with a decreased rate until the second day after treatment. The production of eggs rate was also decreased until the second day after treatment in all 3 tested groups. Therefore, the association between deltamethrin and B. bassiana and M. anisopliae could be used for B. microplus integrated management. Based on compatibility between acaricide and biological agents, it could be possible the use of both simultaneously on infested cattle. Thus, it is possible to control ticks resistant to deltamethrin if an association with entomogenous fungi such as Beauveria bassiana and Metarhizium anisopliae is established. The best performance of these fungi, associated or not with deltamethrin, was observed for a short period after treatment. / O principal ectoparasito dos bovinos ? o carrapato Boophilus microplus, que tem como caracter?stica grande capacidade de resistir ?s adversidades do meio ambiente sendo causador de diversas patologias, severos preju?zos econ?micos aos produtores e possui grande capacidade de desenvolver resist?ncia a carrapaticidas qu?micos. As pesquisas realizadas utilizando controle alternativo de B. microplus t?m apontado que os fungos que naturalmente promovem a infec??o e at? a morte de diversas esp?cies e est?gios de vida de carrapatos s?o, promissores candidatos a esse papel de agente de controle microbiano. Com o objetivo de fornecer subs?dios que complementar?o o controle desta parasitose, avaliou-se in vivo a compatibilidade entre a deltametrina e os fungos Beauveria bassiana e Metarhizium anisopliae sobre cepa resistente de B. microplus a deltametrina, atrav?s do teste de est?bulo, com bovinos infestados artificialmente. Foram utilizados oito bezerros, para cada bioensaio, alojados individualmente, infestados em dias alternados com larvas de B. microplus por um per?odo de 25 dias. Ao t?rmino da infesta??o, foram realizados quatro tratamentos com dois animais por grupo. O grupo controle recebeu somente ?gua; o grupo tratado com fungo recebeu suspens?o conidial na concentra??o de 108 con?dios/ml; o grupo tratado com deltametrina recebeu solu??o na concentra??o de 25 ppm e o grupo tratado com associa??o recebeu suspens?o do fungo na concentra??o de 108 con?dios/ml com deltametrina a 25 ppm. Os resultados foram obtidos atrav?s da contagem di?ria dos carrapatos desprendidos do corpo dos animais, al?m da avalia??o dos par?metros biol?gicos das f?meas, as quais foram incubadas em c?mara climatizada. Realizaram-se dois bioensaios, sendo o primeiro com o fungo Metarhizium anisopliae, onde os percentuais m?dios de mortalidade foram: 32,57% para este fungo, 38,58 % para deltametrina e 30,92% para associa??o entre ambos. No segundo bioensaio foi utilizado o fungo Beauveria bassiana, onde os percentuais m?dios de mortalidade foram: 26,06% para este fungo, 64,30% para deltametrina e 70,64% para associa??o entre ambos. Quanto aos par?metros biol?gicos, as principais altera??es em ambos bioensaios foram a redu??o do ?ndice nutricional, principalmente at? o quarto dia ap?s tratamento, com exce??o da deltametrina que reduziu at? o segundo dia ap?s o tratamento. O ?ndice de produ??o de ovos s? foi reduzido at? o segundo dia ap?s tratamento para todos os grupos do primeiro bioensaio e somente no quarto dia ap?s o tratamento para o B. bassiana. Sendo assim, a associa??o entre deltametrina e os fungos testados pode ser utilizada como ferramenta para o manejo integrado de B. microplus, demonstrando compatibilidade in vivo entre acaricida qu?mico e bio l?gico, sendo poss?vel sua utiliza??o, simult?nea sobre bovinos infestados com B. microplus. O melhor desempenho desses fungos, associados ou n?o, foi observado em um per?odo curto ap?s o tratamento.

Made available in DSpace on 2016-04-28T20:16:21Z (GMT). No. of bitstreams: 1 2007-Everton Kort Kamp Fernandes.pdf: 2844359 bytes, checksum: a56e9f806dc8f1f0e7cc6b4641b4a78c (MD5) Previous issue date: 2007-03-30 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Beauveria bassiana is an entomopathogenic fungus used worldwide as a biological control agent of pest arthropods. The present study aimed at characterizing several isolates of B. bassiana and selects those with major potential to control ticks population and other arthropods under natural conditions. Thus, 53 isolates of B. bassiana and 7 isolates including 5 other Beauveria spp. and 1 Engyodontium albus (=Beauveria alba) were investigated. These isolates were originally from different regions of Brazil and other nations, and different hosts or substrate. Initially, all the isolates were morphologically analyzed and investigated about the potential of conidial production. Afterwards, experiments were conducted to investigate the genetic diversity of Brazilian isolates of B. bassiana, to evaluate the spatial distribution of genotypes, and to understand the genetic relationship among isolates of different species of Beauveria. Thus, all isolates were analyzed by isozymes electrophoresis, AFLP-PCR and sequencing of ITS1 and ITS2 regions. The results have permitted the taxonomic identification of isolates and the selection of those with major potential to conidial production. The results have also showed the detection of high genetic variability among B. bassiana isolates, indicating that this fungus represents a species aggregate. Furthermore, the genetic distance among isolates was directly related to the geographic distances. In addition, the isolates were evaluated to UV-B tolerance, heat tolerance, and cold activity. Heat and cold are two abiotic factors of the environment that restrict the use of entomopathogenic fungi as agents for biological control of insects. In addition, solar radiation, particularly the UV-B wavelength, is a major environmental factor that can negatively affect this entomopathogenic fungus in the field. High variability in UV-B tolerance was detected among the isolates after 2 h of radiation exposure. Conidial relative germination ranged from 0% to almost 80%, and a delayed germination was detected. Moreover, isolates of B. bassiana originating from lower latitudes tended to have lower UV-B tolerance than isolates from higher latitudes. A high variability in thermotolerance was also observed among the isolates after 2 h of heat exposure at 45 ?C, ranging from low, to medium, to high viability. The thermal death point of most resistant isolates was between 44 ?C and 45 ?C after 6 h of heat exposure. At cold temperatures most of the isolates grew at 5 ?C. An attempt to correlate the latitude of origin with thermotolerance or cold activity indicated that isolates from higher latitudes were more cold-active than isolates from nearer the equator. The present study also evaluated the virulence of the isolates toward Boophilus microplus larvae, and virulence ranged from very low to very high. It was observed that different populations of B. microplus may present different susceptibility to B. bassiana infection. Therefore, the present study has selected isolates of this entomopathogenic fungus with major effective potential to control B. microplus and other pest arthropods under environmental conditions. / Beauveria bassiana ? um fungo entomopatog?nico utilizado em v?rias partes do mundo como agente de controle biol?gico de artr?podes. Baseado nisso, o presente estudo buscou caracterizar diversos isolados de B. bassiana e selecionar aqueles com maior potencial para controlar popula??es de carrapatos e outros artr?podes em condi??es naturais. Neste sentido, 53 isolados de B. bassiana e mais sete isolados incluindo cinco esp?cies de Beauveria spp. e uma de Engyodontium albus (=Beauveria alba) foram estudadas. Esses isolados foram oriundos de v?rias regi?es geogr?ficas brasileiras e estrangeiras, e de diferentes hospedeiros ou substratos. Os isolados foram inicialmente avaliados morfologicamente e quanto ao potencial de produ??o de con?dios. Em seguida, buscou-se investigar a diversidade gen?tica dos isolados brasileiros de B. bassiana, avaliar a distribui??o espacial de gen?tipos, e conhecer a rela??o gen?tica entre os isolados das diferentes esp?cies. Para esta finalidade os isolados foram submetidos as t?cnicas de eletroforese de isoenzimas, AFLP-PCR e seq?enciamento de regi?es ITS1 e ITS2. Os resultados permitiram a confirma??o taxon?mica dos isolados e a sele??o daqueles com maior capacidade de produ??o de con?dios. Permitiram tamb?m detectar grande variabilidade gen?tica entre os isolados de B. bassiana indicando que este fungo representa um agregado de esp?cies. Al?m disso, a dist?ncia gen?tica detectada entre os isolados foi fortemente relacionada a dist?ncia geogr?fica de origem dos isolados. Posteriormente, os isolados tamb?m foram avaliados quanto a toler?ncia ? radia??o UV-B, toler?ncia ao calor e atividade ao frio. Tanto o calor quanto o frio s?o fatores abi?ticos ambientais que restringem o uso de fungos como agentes de biocontrole, enquanto a radia??o solar, particularmente UV-B, ? o maior fator ambiental que pode afetar negativamente a a??o deste entomopat?geno. Neste sentido, foi observada alta variabilidade entre os isolados analisados em rela??o ? toler?ncia a UV-B ap?s duas horas de exposi??o ? radia??o, obtendose percentuais de germina??o de con?dios que variaram entre 0% e aproximadamente 80%. Al?m disso, foi observado atraso na germina??o dos con?dios, e revelado que os isolados originados de baixas latitudes tenderam a apresentar menor toler?nc ia a UV-B do que aqueles provenientes de latitudes mais elevadas. Foi tamb?m encontrada alta variabilidade em termotoler?ncia dos con?dios ap?s duas horas de exposi??o a 45 ?C, variando entre baixa, m?dia e alta viabilidade. Os con?dios mais resistentes n?o toleraram exposi??o entre 44 ?C e 45 ?C por seis horas. Quando submetidos a 5 ?C a maioria dos isolados apresentaram atividade. Os isolados de B. bassiana obtidos de latitudes mais elevadas foram mais ativos ao frio do que aqueles originados pr?ximos ? linha do equador. Al?m disso, os isolados tamb?m foram avaliados quanto ao potencial virulento sobre larvas de Boophilus microplus, e neste sentido foi poss?vel diferenci?-los em baixa, m?dia e alta virul?ncia. Foi detectado tamb?m que diferentes popula??es desta esp?cie de carrapato podem manifestar diferentes n?veis de suscetibilidade ? infec??o por B. bassiana. Neste sentido, o presente estudo permitiu selecionar isolados deste entomopat?geno com maior potencial para o controle biol?gico de B. microplus e outros artr?podes em condi??es de campo.

Made available in DSpace on 2016-04-28T20:16:27Z (GMT). No. of bitstreams: 1 2006-Sandra Borges da Silva.pdf: 1572214 bytes, checksum: 1980363c45abbde9170747be40d0635f (MD5) Previous issue date: 2006-02-24 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Biological control of the tick Boophilus microplus with Metarhizium anisopliae and Beauveria bassiana has been evaluated by several researchers. The selection of specimens resistant to chemical products is a mechanism used by arthropods for survival and maintenance in the environment. It is already known how fungi penetrate the host and their pathogenic potential at different phases of their biological cycle, however, ticks immune response against these agents need more detailed studies. The present work had as objectives: to identify and quantify the cellular types involved in the cellular response of B. microplus inoculated with entomopathogenic (M. anisopliae and B. bassiana) and non-entomopathogenic fungi (Penicillium corylophilum and Fusarium oxysporum). In this study 60 engorged females of B. microplus were used, representing six treatment groups with 10 specimens each. Ticks were inoculated with aqueous suspension of conidia. There were two control groups: in the first one ticks were inoculated with Tween 80 0,1% aqueous solution (negative control), in the second one ticks were not inoculated (testimony group). Fungi suspension or Tween 80 solution were applied in the back part of the idiosoma of the tick. The hemolinfa samples were collected during the whole period of life of the specimens, beginning 24 hours after inoculation. The haemolymph samples were fixed in methanol and stained with Giemsa. In all studied periods, cells like pro-hemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids were observed in the specimens of all groups. Prohemocytes, plasmatocytes and spherulocytes were the most numerous cells observed in the hemolinfa. Hemocytes were absent in the group inoculated with B. bassiana 72 hours after inoculation. After this period, specimens inoculated with entomopathogenic fungi were dead. The absence of cells suggests the immune-suppressor effect of the fungi on this tick species. It was not observed when ticks were treated with M. anisopliae due to accentuated mortality caused by this fungus species. The nonentomopathogenic fungi did not affect the life cycle of this tick, being quickly eliminated of the organism. It was suggested based on the absence of conidia during sampling. / O controle biol?gico do carrapato Boophilus microplus com a utiliza??o de Metarhizium anisopliae e Beauveria bassiana tem sido avaliado por diversos pesquisadores. A sele??o de cepas resistentes ? um mecanismo adotado pelos artr?podes para sobreviv?ncia da esp?cie e sua manuten??o no ambiente. Apesar de estar esclarecida a forma como o fungo penetra no hospedeiro e comprovada sua patogenicidade sobre diferentes fases de seu ciclo biol?gico, a sua resposta imune frente a estes agentes agressores necessita de maiores estudos. O presente trabalho teve como objetivo: identificar e quantificar os tipos celulares envolvidos na resposta imune celular de B. microplus inoculados com fungos entomopatog?nicos n?o entomopatog?nicos. Na experimenta??o foram utilizados 60 f?meas ingurgitadas de Boophilus microplus, representando seis tratamentos cada um contendo 10 esp?cimes. Para os grupos inoculados com fungos entomopatog?nicos foi utilizada a suspens?o aquosa dos fungos Metarhizium anisopliae (isolado Ma 959) e Beauveria bassiana (isolado Bb 986). Nos tratamentos com fungos n?o entomopatog?nicos, Penicillium corylophilum e Fusarium oxysporum, foram formados ainda um grupo testemunha (n?o recebeu inocula??o) e um grupo inoculado com solu??o de tween 80 a 0,1% em ?gua destilada est?ril, considerado controle negativo. Nos grupos dos tratamentos com suspens?o f?ngica ou solu??o a inocula??o foi feita na regi?o posterior do idiossoma do carrapato. As coletas de hemolinfa foram realizadas durante todo o per?odo de vida dos esp?cimes, tendo inicio 24 horas ap?s inocula??o. As amostras de hemolinfa foram fixadas com metanol e coradas com Giemsa. Em todos os per?odos estudados, tanto nos esp?cimes inoculados com fungos como nos controles, foram observados pr?-hem?citos, plasmat?citos, granul?citos, esferul?citos e oenocit?ides. Pr?-hem?citos, plasmat?citos e esferul?citos foram ?s c?lulas mais numerosas na hemolinfa. Foi observada a aus?ncia de hem?citos no grupo inoculado com B. bassiana 72 horas ap?s a inocula??o, como tamb?m a morte dos esp?cimes inoculados com fungos entomopatog?nicos a partir deste per?odo. A aus?ncia de c?lulas evid?ncia o efeito imunossupressor do fungo sobre os carrapatos estudados. Caracter?stica n?o observada para Metarhizium anisopliae, mesmo tendo provocado acentuada mortalidade dos carrapatos. Os fungos n?o entomopatog?nicos n?o afetaram de forma significativa o ciclo de vida destes carrapatos, sendo rapidamente eliminados do organismo.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-06-09T16:27:04Z No. of bitstreams: 1 ISABELE DA COSTA ANGELO.pdf: 4140152 bytes, checksum: bbd35489ea24b040be77df36cd797987 (MD5) / Made available in DSpace on 2016-06-09T16:27:32Z (GMT). No. of bitstreams: 1 ISABELE DA COSTA ANGELO.pdf: 4140152 bytes, checksum: bbd35489ea24b040be77df36cd797987 (MD5) Previous issue date: 2011-03-03 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The study evaluated the protein and lipid profiles of Rhipicephalus microplus engorged females after infection by Metarhizium anisopliae, Beauveria bassiana and Fusarium oxysporum. The treatments were immersion or inoculation of conidial suspension in R. microplus. The hemolymph was collected 24 and 48 hours after treatment. The cell-free hemolymph was separated of hemocytes by centrifugation and hemocytes resuspended in phosphate buffer pH 7.2. The amount of total protein was determined in both fractions of hemolymph and hemocytes were quantified. The cell-free hemolymph was filtered through a 100 kDa and 10 kDa membranes, and analyzed by electrophoresis and liquid chromatography (HPLC). The proteome of cell-free hemolymph (treatment by injection) was evaluated by 2DPAGE. Changes were observed in amount total protein and the amount of hemocytes, but no difference was observed in the electrophoretic profile (1D-PAGE) of the cell-free hemolymph. In haemocytes, the entomopathogens reduced the amount of serpins, while F. oxysporum caused increased. In 2D-PAGE variations were observed in both expression and presence/absence of protein between the groups. The cell-free hemolymph antimicrobial activity was tested against Escherichia coli and Staphylococcus aureus and against the fungus used in the treatment of engorged ticks. The hemolymph collected 48 hours after the immersion treatment with B. bassiana apresented activity anti-B. bassiana with 48 hours of evaluation. This hemolymph was subjected to Superose column to HPLC and peak was collected and analyzed on the analytical column C18. The fractions were collected from the C18 and its apresented activity anti-B. bassiana, but showed no activity against Candida albicans. These fractions were analyzed by Maldi-Tof and most of them had in common an ion with m/z 1,119.5; however, other ions may be involved with this activity antimicrobial. The lipids present in cell-free hemolymph, in the hemocyte and fat body were extracted and analyzed by thin layer chromatography (TLC) or HPTLC for neutral lipids and phospholipids. The classes of neutral lipids in the cell-free hemolymph were cholesterol ester, cholesterol (CHO) and fatty acids (FA), which have varied depending on the fungus used, type of treatment and observation time. Phospholipids found were phosphatidylcholine and phosphatidylethanolamine, its were not significantly altered after the fungal infection. In hemocytes, the same classes of lipids were found and B. bassiana modified phospholipids, while M. anisopliae s.l. altered FA and CHO. The fat body showed, in addition to these classes of neutral lipids, the triglycerides, which increased significantly 48 hours after inoculation with M. anisopliae s.l. The lipase activity in fat body was measured and it was demonstrated that increased activity 48 hours after inoculation, mainly in the group inoculated with Metarhizium. Therefore, the results showed alterations related to the proteins expression in the hemocytes and the cell-free hemolymph after inoculation with fungi, immunosuppression of hemocytes and antimicrobial peptides induction after infection with B. bassiana, besides changes in the lipid profile of R. microplus after infection. However, further studies are necessary to understand these changes. / O trabalho avaliou os perfis prot?ico e lip?dico de f?meas ingurgitadas de Rhipicephalus microplus ap?s infec??o com Metarhizium anisopliae s.l., Beauveria bassiana ou Fusarium oxysporum. Os tratamentos foram imers?o ou inocula??o da suspens?o conidial em R. microplus. A hemolinfa foi coletada 24 e 48 horas ap?s os tratamentos. O plasma foi separado dos hem?citos por centrifuga??o e os hem?citos resuspensos em tamp?o fosfato pH 7,2. A concentra??o de prote?na total foi determinada em ambas as fra??es da hemolinfa e os hem?citos quantificados. O plasma da hemolinfa foi filtrado em membrana de 100 kDa e 10 kDa, sendo analisados por eletroforese e cromatografia l?quida de alta efici?ncia (CLAE). O proteoma do plasma da hemolinfa (tratamento por inocula??o) foi avaliado por gel 2D. Foram observadas varia??es na quantidade de prote?na total de ambas as fra??es da hemolinfa, na quantidade de hem?citos bem como na intensidade de prote?nas/pept?deos expressos no plasma da hemolinfa. Nos hem?citos, os entomopat?genos reduziram a quantidade de serpinas, enquanto F. oxysporum causou aumento. No gel 2D foram observadas varia??es na express?o bem como na aus?ncia/presen?a de prote?nas entre os grupos. O plasma da hemolinfa teve sua atividade antimicrobiana testada contra Escherichia coli e Staphylococcus aureus e contra o fungo utilizado no tratamento das f?meas ingurgitadas. A hemolinfa coletada 48 horas ap?s o tratamento por imers?o com B. bassiana apresentou atividade anti-B. bassiana com 48 horas de avalia??o. Esta hemolinfa foi submetida ? coluna Superose de CLAE e o pico coletado analisado na coluna anal?tica C18. As fra??es coletadas da C18 apresentaram atividade anti-B. bassiana, por?m n?o apresentaram atividade contra Candida albicans. Essas fra??es foram analisadas por Maldi-Tof e a maioria delas apresentou um ?on com raz?o m/z 1.119,5; no entanto, outros ?ons podem estar envolvidos com essa atividade antimicrobiana. Os lip?deos presentes no plasma da hemolinfa, nos hem?citos e no corpo gorduroso foram extra?dos e analisados por cromatografia em camada delgada (CCD) ou CCD de alta performance para lip?deos neutros e fosfolip?deos. As classes de lip?deos neutros encontradas no plasma da hemolinfa foram colesterol-?ster, colesterol (CHO) e ?cidos graxos (AG), que sofreram altera??es em fun??o do fungo utilizado, tipo de tratamento e tempo de observa??o. Os fosfolip?deos encontrados foram fosfatidilcolina e fosfatidiletanolamina, que n?o foram significativamente alterados ap?s a infec??o f?ngica. Nos hem?citos, as mesmas classes de lip?deos foram encontradas e B. bassiana alterou os fosfolip?deos, enquanto M. anisopliae s.l. alterou os AG e CHO. O corpo gorduroso apresentou, al?m destas classes de lip?deos neutros, o triacilglicerol, que aumentou significativamente 48 horas ap?s a inocula??o com M. anisopliae s.l. A atividade lipase no corpo gorduroso foi mensurada, sendo evidenciado um aumento 48 horas ap?s a inocula??o, principalmente no grupo inoculado com Metarhizium. Portanto, os resultados demonstraram altera??es na express?o de prote?nas no plasma da hemolinfa e nos hem?citos ap?s inocula??o com os fungos, imunossupress?o dos hem?citos, indu??o de pept?deos com atividade antimicrobiana ap?s infec??o com B. bassiana, al?m de altera??es no perfil lip?dico de R. microplus ap?s infec??o. No entanto, maiores estudos s?o necess?rios para o entendimento dessas altera??es.

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-08-29T16:13:08Z No. of bitstreams: 1 2010 - Paula Sant?ana Alves.pdf: 1049254 bytes, checksum: 295fd608eecdae86916f71cbebfc389e (MD5) / Made available in DSpace on 2016-08-29T16:13:08Z (GMT). No. of bitstreams: 1 2010 - Paula Sant?ana Alves.pdf: 1049254 bytes, checksum: 295fd608eecdae86916f71cbebfc389e (MD5) Previous issue date: 2016-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPQ / Stomoxys calcitrans is a hematophagous fly of which parasitism causes economic losses related to the reduction of milk production and weight gain. Due to the high level of resistance of the stable fly to most of chemical insecticides, it is necessary to search for new alternatives for their control, and one of them would be the use of entomopathogenic fungi. However, few researches related to microbial control of this fly were carried out. The objective of this study was to assess the ability of the Lecanicillium lecanii fungus in to make unfeasible eggs, larvae and pupae of the stable fly under laboratory conditions and verify the formation of inhibition zone in agar solid due to the presence of substances in the mucus and total homogenized larvae not previously exposed and sensitized to the fungus. The S. calcitrans colony was maintained in the Laborat?rio de D?pteros Hemat?fagos, UFRRJ. The tests were performed in the Laborat?rio de Controle Microbiano de Artr?podes, UFRRJ, where the strain CG 420 of L. lecanii was used to prepare the water suspensions. In assays with eggs, larvae and pupae of S. calcitrans, fungal suspensions were used at 2x108, 2x107, 2x106, 2x105 con.mL-1 concentrations, besides the positive control. In assays with eggs, they were immersed in different fungal suspension and transferred to Petri dishes with paper filter, which were added larval rearding medium and its respective suspension. Mortality was assessed five days after exposure, where it was observed whether stage change occurred. There was no significant difference in the hatching of eggs exposed to different fungal concentrations against the control. Larvae were subjected to the same treatment of eggs, and mortality was assessed ten to fifteen days after exposure. There was no significant difference in mortality of larvae exposed to the fungus when compared with the control group, however, the fungus L. lecanii was significantly able to reduce adult emergence from larvae exposed to the highest tested concentration and there was a percentage of emergence of 25,45% and 43,64% compared to 63,64% and 90,91% of the control group. The pupae were also immersed in different fungal suspensions, and transferred to Petri dishes with paper filter, added to its respective suspension. Mortality was assessed ten days after exposure. There was no significant difference in mortality of pupae exposed to different fungi concentrations. Filter paper discs were soaked in mucus samples and total homogenized from larvae of S. calcitrans not exposed and previously sensitized to the fungus L. lecanii and placed in petri dishes sown with the same fungus. It was not observed the formation of inhibition zone around the discs. The L. lecanii fungus was unable to make unfeasible eggs, larvae or pupae of S. calcitrans in tested fungal concentrations, however, was significantly able to reduce the emergence of adults when their third larvae stage were exposed to 2x108 con.mL-1 concentration. / Stomoxys calcitrans ? um d?ptero hemat?fago cujo parasitismo ocasiona perdas econ?micas relacionadas ? redu??o da produ??o leiteira e ganho de peso, al?m de estar associado ? dissemina??o e transmiss?o de agentes patog?nicos. Devido ao elevado grau de resist?ncia da mosca dos est?bulos a uma diversidade de inseticidas qu?micos, torna-se necess?rio a busca por alternativas para o seu controle, e uma delas seria a utiliza??o de fungos entomopatog?nicos. Todavia, poucas pesquisas relacionadas ao controle microbiano desta mosca foram realizadas. O objetivo deste estudo foi verificar a capacidade do fungo Lecanicillium lecanii em inviabilizar ovos, larvas e pupas de S. calcitrans em condi??es laboratoriais e verificar a forma??o de halo de inibi??o em meio de cultivo s?lido, em decorr?ncia de subst?ncias presentes no muco e no macerado total de larvas n?o expostas e previamente sensibilizadas ao fungo. A col?nia de S. calcitrans foi mantida no Laborat?rio de D?pteros Hemat?fagos, UFRRJ. Os ensaios foram realizados no Laborat?rio de Controle Microbiano de Artr?podes, UFRRJ. Nos ensaios foram utilizadas suspens?es aquosas do isolado CG 420 de L. lecanii nas concentra??es de 2x108, 2x107, 2x106, 2x105 con.mL-1, al?m do controle positivo. Nos ensaios com ovos, estes foram imersos nas suspens?es f?ngicas, e transferidos para placas de Petri com papel filtro, meio de desenvolvimento larval e 3 mL de sua respectiva suspens?o. A mortalidade foi avaliada cinco dias ap?s a exposi??o. N?o foi observada diferen?a significativa na eclos?o dos ovos tratados frente ao controle. As larvas foram submetidas ao mesmo tratamento dos ovos, e a mortalidade foi avaliada dez e quinze dias ap?s a exposi??o. N?o foi observada diferen?a significativa na mortalidade de larvas expostas ao fungo quando comparados com o grupo controle, entretanto, o fungo L. lecanii reduziu significativamente a emerg?ncia de adultos provenientes das larvas expostas ? concentra??o m?xima testada (2x108 con.mL-1) sendo observado um percentual de emerg?ncia de 25,45% e 41,82 %, em compara??o com 63,64% e 90,91% do grupo controle. As pupas tamb?m foram imersas nas suspens?es f?ngicas, e transferidas para placas de Petri com papel filtro e sua respectiva suspens?o. A mortalidade foi avaliada dez dias ap?s a exposi??o. N?o foi observada diferen?a significativa na mortalidade das pupas expostas ?s diferentes concentra??es f?ngicas. Discos de papel filtro foram embebidos em amostras de muco e macerado total proveniente de larvas de S. calcitrans n?o expostas e previamente sensibilizadas ao fungo L. lecanii e colocados em placas de Petri semeadas com o mesmo fungo (1x108 con.mL-1). N?o foi verificada a forma??o de halo de inibi??o ao redor dos discos. O fungo L. lecanii foi incapaz de inviabilizar ovos, larvas ou pupas de S. calcitrans nas concentra??es f?ngicas testadas, no entanto, reduziu significativamente a emerg?ncia de adultos quando suas larvas de terceiro est?dio foram expostas a concentra??o 2x108 con.mL-1.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-09-08T12:13:31Z No. of bitstreams: 1 2011 - Jos? Antonio Batista da Silva.pdf: 1901739 bytes, checksum: bf67996decc8c7cef031a8ce05d6f464 (MD5) / Made available in DSpace on 2016-09-08T12:13:31Z (GMT). No. of bitstreams: 1 2011 - Jos? Antonio Batista da Silva.pdf: 1901739 bytes, checksum: bf67996decc8c7cef031a8ce05d6f464 (MD5) Previous issue date: 2011-12-08 / This work was carried out from August 2007 to July 2011, in Itabora?, RJ, Brazil, and aimed to identify the Calliphoridae species exist in a mangrove swamp, verifying which are the predominant species, highlighting the proportions of males and females, and also analyzing the influence of abiotic factors such as temperature, humidity and moon phases, and also providing an analysis of the abundance, richness, diversity and similarity between the periods of sampling, ecological relationships between species of the same family and the ecological relationship between Calliphoridae and Phoridae parasitoids in Guapi-Mirim Environmental Protection Area, (Guapi-Mirim EPA) in the same city. Semiweekly collections were carried out over 48 months (96 collections). The traps were suspended at a height of 1.20 m above the ground for a period of 48 hours for each collection. To trap and collect the flies, four plastic traps (35 cm x 15 cm) were placed 100m apart each other in the studied area. The bait used was based on fish (sardine) in decomposition. The insects caught were killed by asphyxiation with 70% ethanol inside the trap. After asphyxiation all specimens were put into plastic pots containing 70% ethanol. The specimens were taken to the Laborat?rio de Transmissores de Leishmanioses (Setor de Entomologia M?dica e Forense) - IOC/FIOCRUZ, RJ, where they were separated by the collection day, counted and then identified using a stereoscopic microscope and the dichotomous keys for families and species. The abundance of Calliphoridae flies was statistically analyzed using the Statistica 7.1 program (StatSoft 2005) for the non-parametric test Kruskal-Wallis one way ANOVA (p< 0.05). Also the Kendall Tau Correlation test, Mann-Whitney (significance of 95%; p<0.05) and chi-square test (?2) were used to analyze. The Shannon-Wiener diversity index, Bray-Curtis similarity and Euclidean Distance were used. A total of 4,531flies were collected. These flies belonged to ten (10) species of the Calliphoridae family: Chrysomya megacephala (Fabricius, 1794) (86.40%), Chrysomya albiceps (Wiedemann,1819) (5.72%), Cochliomyia macellaria (Fabricius, 1775) (4.94%), Chrysomya putoria (Wiedemann, 1818) (2.10%), Lucilia eximia (Wiedemann, 1819) (0.30%), Cochliomyia hominivorax (Cocquerel, 1858) (0.22%), Chloroprocta idioidea (Robineau-Desvoidy, 1930) (0.20%), Hemilucilia segmentaria (Fabricius, 1805) (0.04%), Hemilucilia semidiaphana (Rondani, 1850) (0.04%), Lucilia cuprina (Wiedemann,1830) (0.04%). The species C. megacephala was the one that presented the greatest abundance. Spring was the preferred season for all species; temperatures between 30.5 and 32.40C and relative humidity between 56.8 and 61.7% were considered those where there was a frenzy of oviposition; the full and new moons were the phases during which there was a higher occurrence of flies; Megaselia scalaris occurred more frequently in the most abundant Calliphoridae species in the same season of the year thus allowing greater dispersion of eggs; Calliphoridae species had a higher rate survival with increased food resources resulting from the polluting activity / Este trabalho foi realizado entre agosto de 2007 e julho de 2011, no Munic?pio de Itabora?, RJ, Brasil, e teve como objetivo identificar as esp?cies de Calliphoridae existentes em uma ?rea de mangue, quantificar as predominantes, destacando as propor??es de machos e f?meas, e tamb?m analisar a influ?ncia de fatores abi?ticos, tais como temperatura, umidade do ar e fases lunares; e ainda relacionar a abund?ncia, a riqueza, a diversidade e a similaridade entre os per?odos de coletas, rela??es ecol?gicas entre as esp?cies da entomofauna da mesma fam?lia e a rela??o ecol?gica entre Calliphoridae e Phoridae parasit?ides dentro da ?rea de Prote??o Ambiental de Guapi-Mirim (APA de Guapi-Mirim), em Itabora?. Durante 48 meses, foram realizadas 96 coletas, uma a cada 15 dias. Em todas as capturas, foram utilizadas armadilhas confeccionadas em recipiente pl?stico com 35 cm de altura e 15 cm de di?metro. Cada uma das quatro armadilhas foi suspensa a uma altura de 1,20 m do solo contendo 100g de isca de peixe (sardinha) em decomposi??o por um per?odo de 48 horas. Ap?s cada captura, todos os esp?cimes foram mortos no interior das armadilhas por asfixia utilizando etanol 70%. Em seguida todos os indiv?duos foram acondicionados em potes pl?sticos, contendo etanol a 70% e encaminhados ao Laborat?rio de Transmissores de Leishmanioses (Setor de Entomologia M?dica e Forense), IOC-FIOCRUZ, RJ, em seguida todos os esp?cimes foram separados por dia de coleta, identificadas e quantificadas. Para tal procedimento utilizou-se um microsc?pio estereosc?pico e chaves dicot?micas para a identifica??o da fam?lia e das esp?cies. As an?lises estat?sticas foram feitas utilizando o programa estat?stico Statistica 7.1 (STATSOFT, 2005), atrav?s do teste de Kruskal- Wallis one way ANOVA, teste de qui- quadrado (?2), Mann-Whitney e correla??o de Kendall Tau (p<0,05). Foram utilizados os ?ndices de diversidade de Shannon-Wiener e similaridade de Bray-Curtis, assim como a Dist?ncia Euclidiana. Foram capturadas 4531 moscas pertencentes a dez (10) esp?cies da fam?lia Calliphoridae: Chrysomya megacephala (Fabricius, 1794) (86,40%), Chrysomya albiceps (Wiedemann,1819) (5,72%), Cochliomyia macellaria (Fabricius, 1775) (4,94%), Chrysomya putoria (Wiedemann, 1818) (2,10%), Lucilia eximia (Wiedemann, 1819) (0,30%), Cochliomyia hominivorax (Cocquerel, 1858) (0,22%), Chloroprocta idioidea (Robineau-Desvoidy, 1930) (0,20%), Hemilucilia segmentaria (Fabricius, 1805) (0,04%), Hemilucilia semidiaphana (Rondani, 1850) (0,04%), Lucilia cuprina (Wiedemann,1830) (0,04%). A esp?cie C. megacephala foi aquela que apresentou a maior abund?ncia, a primavera foi a esta??o do ano em que todas as esp?cies mostraram maior prefer?ncia; as temperaturas compreendidas entre 30,5 e 32,40C e a umidade relativa do ar entre 56,8 e 61,7% foram consideradas aquelas onde houve frenesi de oviposi??o; nas luas cheias e novas houve maior ocorr?ncia de moscas capturadas; Megaselia scalaris se utilizou de esp?cies abundantes nas mesmas esta??es do anos em que ela ocorreu com maior frequ?ncia, possibilitando assim maior dispers?o de ovos; a maior disponibilidade de recursos alimentares resultantes da atividade poluidora tempor?ria ofereceram uma melhor condi??o de sobreviv?ncia para as esp?cies os Calliphoridae.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-09-09T11:45:13Z No. of bitstreams: 1 2011 - Vanessa de Almeida Raia.pdf: 1415435 bytes, checksum: 6553cdee2bd5a5f2a363cb3cd11f812d (MD5) / Made available in DSpace on 2016-09-09T11:45:13Z (GMT). No. of bitstreams: 1 2011 - Vanessa de Almeida Raia.pdf: 1415435 bytes, checksum: 6553cdee2bd5a5f2a363cb3cd11f812d (MD5) Previous issue date: 2011-01-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / To fill some gaps about the intrinsic mechanisms of the biology of oviposition of R. (B.) microplus, we evaluated the concentration of lipids and carbohydrates, as well as the variability of lipid in eggs per day of posture. For this, engorged females weighing between 151 and 360 mg were placed in controlled environment (27 ? 1 ? C, 80 ? 5% RH, darkness), and after beginning of laying, three samples of the eggs pool were daily collected, packed and preserved at -20 ? C. Samples 1 and 2, weighing 50 mg each, were used to quantitate lipids and glucose, and to evaluate the lipid profile, respectively; and the third sample, containing 1 g of eggs, was used to quantitate glycogen. The determination of total lipids was performed using phospho-vanillin reagent, while the concentrations of cholesterol, triglycerides and glucose were obtained by colorimetric determination using endpoint enzymatic kit. The level of glycogen was performed using the acid 3,5 dinitrosalicylic reactive . The results were analyzed by nonparametric Kruskal-Wallis and the relationship between the concentrations of substrates and the days of laying was assessed by polynomial regression. There was no significant difference in the concentrations of these substrates in eggs from different days of oviposition, as there was no relationship of these concentrations with the days of laying. After lipid extraction, the samples were performed in thin layer chromatography, high-performance (HPTLC) for neutral lipids and phospholipids. Then, densitometry was performed using the Image Master Total Lab and the daily percentage of each lipid in the samples was calculated. Among the neutral lipids were identified the hydrocarbons, esterified cholesterol, triglycerides, fatty acids, free cholesterol and diacylglycerol. Of these, the cholesterol ester showed the highest variation, it was not detected in eggs at the fifth day of oviposition. Among the phospholipids were detected the phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine and sphingomyelin fosfatidilinusitol. This latter was absent in eggs of the seventh day onwards. Thus, the different rates of hatchability of eggs from different laying days cannot be attributed to the concentrations of total lipids, although some specific lipids such as cholesterol ester and sphingomyelin might influence the difference in larval hatchability, more researches are needed for clarify the role of these substrates in embryogenesis / Objetivando preencher algumas lacunas sobre os mecanismos intr?nsecos da biologia da oviposi??o de R. (B.) microplus, foram avaliadas as concentra??es de lip?dios e carboidratos, assim como a variabilidade lip?dica dos ovos postos em diferentes dias de postura. Para isso, f?meas ingurgitadas pesando entre 151 e 360 mg foram colocadas em estufa biol?gica sob condi??es controladas (27 ? 1oC, 80 ? 5% UR, escotofase), e ap?s in?cio da postura, tr?s amostras di?rias do ?pool? de ovos foram coletadas, acondicionadas e preservadas a ?20?C. As amostras 1 e 2, pesando 50 mg cada, foram utilizadas para dosar lip?dios e glicose, e avaliar o perfil de lip?dios, respectivamente; a amostra 3, contendo 1 g de ovos foi utilizada para dosar glicog?nio. A dosagem de lip?dios totais foi realizada atrav?s do reagente de fosfo-vanilina, enquanto as concentra??es de colesterol, triglicer?dios e glicose foram obtidas atrav?s de determina??o colorim?trica com a utiliza??o de kit enzim?tico de ponto final. A dosagem de glicog?nio foi realizada atrav?s do reativo ?cido 3,5 dinitrosalic?lico. Os resultados das dosagens foram analisados pelo teste n?o param?trico de Kruskal-Wallis e a rela??o entre as concentra??es dos substratos e os dias de postura foi avaliada atrav?s de Regress?o polinomial. N?o foram constatadas diferen?as significativas entre as concentra??es destes substratos nos ovos de diferentes dias de oviposi??o, assim como n?o houve rela??o dessas concentra??es com os dias de postura. Ap?s extra??o dos lip?dios das amostras, foram realizadas cromatografias em camada delgada de alto desempenho (HPTLC) para lip?dios neutros e fosfolip?dios. Em seguida, a densitometria foi realizada atrav?s do programa Image Master Total Lab e o percentual di?rio de cada lip?dio nas amostras foi calculado. Dentre os lip?dios neutros foram identificados hidrocarbonetos, colesterol esterificado, triglicer?dios, ?cidos graxos, colesterol livre e diacilglicerol. Destes, o colesterol esterificado foi o que apresentou maior varia??o, n?o sendo detectado nos ovos do quinto dia de oviposi??o. Dentre os fosfolip?dios foram detectados o ?cido fosfat?dico, fosfatidiletanolamina, fosfatidilcolina, fosfatidilinusitol e esfingomielina. Sendo este ?ltimo ausente nos ovos do s?timo dia de postura em diante. Assim, as diferentes taxas de eclodibilidade por dia de postura n?o podem ser atribu?das ?s concentra??es de lip?dios totais, embora alguns lip?dios espec?ficos como o colesterol esterificado e a esfingomielina possam ter influ?ncia na diferen?a de eclodibilidade larval, sendo necess?rios mais estudos para eclarecer o papel desses substratos na embriog?nese

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-09-14T14:46:35Z No. of bitstreams: 1 2011 - Jurema Schinz Diniz.pdf: 1784426 bytes, checksum: ef765e40d98144beee9ab74d12aff5f8 (MD5) / Made available in DSpace on 2016-09-14T14:46:35Z (GMT). No. of bitstreams: 1 2011 - Jurema Schinz Diniz.pdf: 1784426 bytes, checksum: ef765e40d98144beee9ab74d12aff5f8 (MD5) Previous issue date: 2011-06-02 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / This study contributes to understanding the relationship between tropical dry forests regeneration and arbuscular mycorrhizal fungi activity. The specific objective of this study is to extract and quantify glomalin, glomerospores and humic substances in different sucessional stages of a dry forest. The main objective is to study the activity of AMFs in different stages of a dry forest and to colabore to the projects: ?Functional links between aboveground changes and belowground activity with land use in the Americas: Soil biodiversity and food security? e ?Human, Ecological and Biophysical Dimensions of Tropical Dry Forest?. Total Glomalin (TG) and carbon levels were higher in inicial and late stage of dry forest, indicating the carbon storage contribution to glomalin in soil. In addition, the higher number of glomerospores in this sucessional stage shows the more activity of AMF and its potential to regeneration of disturbed dry forests. The different chemical and physical properties of soil in intermediate stage possibly contributed to low glomalin and carbon levels. The higher levels of humic substances in this stage possibly contribute to the lower activity of AMF and can be explained by the higher diversity of plants in this area. It is important to study the influence of physical and chemical properties, humic substances and plant diversity in AMF activity, in different sucessional stages of dry forests. / Este trabalho contribui para o entendimento da rela??o da regenera??o de florestas tropicais secas, com a atividade dos fungos micorr?zicos arbusculares (FMAs). Tem como objetivos espec?ficos: a extra??o e a quantifica??o de glomalina, de glomerosporos e de subst?ncias h?micas em diferentes sucess?es de floresta seca. O objetivo ? avaliar a atividade dos FMAs em diferentes est?gios sucessionais de floresta seca, acrescentando e colaborando assim para o desenvolvimento dos projetos: ?Functional links between aboveground changes and belowground activity with land use in the Americas: Soil biodiversity and food security? e ?Human, Ecological and Biophysical Dimensions of Tropical Dry Forest?. Os teores de glomalina total e de carbono org?nico foram maiores na ?rea de sucess?o inicial e tardia de floresta seca, indicando maior influ?ncia do estoque de carbono na glomalina do solo. O maior n?mero de esporos e glomalina, na ?rea de sucess?o inicial, mostrou a maior atividade dos fungos micorr?zicos arbusculares em ?rea perturbada que est? se regenerando, sugerindo a contribui??o dos FMAs para essa recupera??o do ecossistema de florestas secas. As propriedades f?sicas e qu?micas diferentes do solo na ?rea de sucess?o intermedi?ria provavelmente contribu?ram para a baixa concentra??o de glomalina e carbono. Os maiores teores de ?cidos h?micos e f?lvicos no solo de sucess?o intermedi?ria podem ter contribu?do tamb?m para a baixa atividade dos FMAs e podem ser devidos ? maior diversidade de esp?cies vegetais nesta ?rea. ? importante desenvolver estudos que comparem as propriedades qu?micas e f?sicas do solo com a atividade desses microorganismos simbiontes em diferentes sucess?es vegetais; que analisem a influ?ncia das subst?ncias h?micas na atividade dos FMAs em campo; e da influ?ncia da diversidade de esp?cies vegetais na atividade dos FMAs.

Submitted by Leticia Schettini (leticia@ufrrj.br) on 2016-09-20T14:14:37Z No. of bitstreams: 1 2016 - NAIARA DE MIRANDA BENTO RODRIGUES.pdf: 1508318 bytes, checksum: bbf2aebb5d43033339bf559d73461919 (MD5) / Made available in DSpace on 2016-09-20T14:14:37Z (GMT). No. of bitstreams: 1 2016 - NAIARA DE MIRANDA BENTO RODRIGUES.pdf: 1508318 bytes, checksum: bbf2aebb5d43033339bf559d73461919 (MD5) Previous issue date: 2016-02-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPQ / Mastitis adversely affects milk production and in general cows do not regain their full production levels post recovery, leading to considerable economic losses. Moreover the percentage decrease in milk production depends on the specific pathogen that caused the infection and enterobacteria are responsible for this greater reduction. These microorganisms are preferentially found in the habitat of animals in places contaminated with feces, urine, clay and also organic beds. Phenotypic tests are among the currently available methods used worldwide to identify enterobacteria; however they tend to misdiagnose the species despite the multiple tests carried out and they can delay the antibiotic therapy by clinic veterinary. On the other hand the MALDI-TOF MS technique has been attracting attention for its precise identification of several microorganisms at species level. In the current study, 183 enterobacteria were detected in milk (n=47) and fecal samples (n=94) collected from cows; also water (n=23) and milk line samples (n=19) collected from a farm in Rio de Janeiro with the purpose to present the MALDI-TOF MS technique as efficient methodology and also as a ?gold standard? to better understand the possible current biochemical errors in enterobacteria identification considering isolates from bovine environments. This proteomic technique confirmed 92.9% (170/183) of the enterobacteria species identified by biochemical tests that showed high sensitivity (> 81%) and specificity (> 89%). The gyrB sequencing was made in eigth from thirteen misidentified enterobacteria and confirmed 100% the MALDI-TOF results, so the proteomic technique was used as a ?gold standard? for this study. The amino acid decarboxylation test made the most misidentifications and Enterobacter spp was the largest misidentified genus (76.9%, 10/13). E.coli was prevalent (83%, 152/183) in all samples and the bovine milk presented the most enterobacteria diversity. The Salmonella sp wasn?t detected in feces bovine samples and all water samples from different points in the farm presented unacceptable microbiological standards. Was identified enterobacteria in milkers hands and nasal cavity also in the milking machines used on the property. These results aim to contribute significantly to the characterization of the Enterobacteriaceae as well in understanding of its spread in dairy production environment , assisting in need diagnostic of possible agents involved in bovine mastitis as well as to implement properly targeted prophylactic measures. / A mastite bovina afeta negativamente a produ??o de leite dificultando a recupera??o dos n?veis de produ??o total das propriedades leiteiras, levando a perdas econ?micas consider?veis. Esta redu??o no percentual da produ??o de leite pode estar associada ao agente patog?nico espec?fico que causou a infec??o, sendo as enterobact?rias frequentemente respons?veis pela mastite ambiental. Estes microrganismos s?o preferencialmente encontrados no habitat normal dos animais como locais que apresentam esterco, urina, barro e camas org?nicas. Os testes fenot?picos est?o entre os m?todos dispon?veis atualmente utilizados para identificar as enterobact?rias; no entanto, eles podem ocasionalmente identitificar erroneamente algumas esp?cies apesar dos m?ltiplos ensaios realizados. Al?m disso, a demora na sua execu??o pode tardar a antibioticoterapia realizada em campo. Por outro lado, a t?cnica de MALDI-TOF MS tem atra?do a aten??o pela sua identifica??o precisa dos v?rios microorganismos em n?vel de esp?cie. No presente estudo, um total de 183 enterobact?rias foram isoladas a partir de amostras de leite (n=47) e fezes colhidas de vacas em lacta??o (n=94); amostras de ?gua (n=23) e na linha de ordenha (n=19) em uma propriedade situada no Rio de Janeiro. A proposta foi utilizar a t?cnica de MALDI-TOF MS como um m?todo eficaz de identifica??o bacteriana de enterobact?rias e descrever a permanencia destes microrganismos no ambiente de produ??o leiteira. A t?cnica prote?mica confirmou 92,9% (170/183) das esp?cies de enterobact?rias identificadas pelos testes bioqu?micos convencionais. O sequenciamento do gene gyrB, realizado em oito das 13 enterobact?rias que apresentaram identifica??o discordante, confirmou em 100% o resultado da t?cnica prote?mica, que foi utilizada como metodologia de refer?ncia no presente estudo. O g?nero Enterobacter foi o mais discordante pelo m?todo bioqu?mico (76,9%, 9/13). A E.coli foi a esp?cie predominante (83%, 152/183) em todas as amostras avaliadas, sendo que o leite bovino apresentou maior diversidade de enterobact?rias. N?o foi detectada a presen?a de Salmonella spp. nas amostras de fezes bovinas e todas as amostras de ?gua dos diferentes pontos de coleta da propriedade apresentaram padr?es microbiol?gicos inaceit?veis. Foram isoladas enterobact?rias das m?os e cavidades nasal dos ordenhadores, bem como nas ordenhadeiras mec?nicas utilizadas na propriedade. Estes dados visam contribuir de forma significativa para a caracteriza??o das enterobacterias bem como para a compreens?o e sua descri??o no ambiente de produ??o leiteira, auxiliando no diagn?stico preciso dos poss?veis agentes envolvidos na mastite bovina bem como na implementa??o de medidas profil?ticas devidamente direcionadas.