Orientador : Ana Maria Lima de Azeredo-Espin / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-02T14:54:27Z (GMT). No. of bitstreams: 1 Lemos_TailaAndrade_M.pdf: 3951909 bytes, checksum: 3f2ba88513abb52f3e6defc4445d311f (MD5) Previous issue date: 2000 / Resuimo: Dermatobia hominis (Linnaus Jr.) conhecida popularmente como mosca do berne. É uma das principais causadoras de miíase primária em vertebrados. sendo uma das principais pragas da pecuária nacional. causando grandes prejuízos à industria coureira. laticínios e frigoríficos. A mosca do berne apresenta uma biologia interessante e ciclo de vida bastante peculiar. o que dificuta a observação da espécie na natureza e a manutenção desta em laboratório. Por estes motivos. a caracterização da variabilidade genética e da estrutura populacional são importantes ferramentas na compreensão da evolução de D. hominis. Para este fim. foram utilizadas as técnicas de RAPD, com seis populações, e de RFLP do DNA mitocondrial, com doze populações. Os resultados foram comparados entre si e com estudos anteriores. Além disso, foram comparadas diferentes abordagens que podem ser aplicadas aos dados obtidos com RAPD. Os resultados de RAPD e RFLP do DNA mitocondrial sugerem que a espécie comporta-se como uma metapopulação. na qual as populações são altamente variáveis e apresentam fluxo gênico entre si. Quanto às diferentes abordagens aplicáveis aos dados de RAPD foi observado que as medidas de Dice e Jaccard, calculadas a partir da matriz completa. e a distância de Nei (1972) calculada com as modificações de Lynch & Milligan (1994), foram as mais adequadas ao estudo da variabilidade de D. hominis. Com o uso do RFLP do mtDNA foram detectados setenta e dois baplótipos, o que denota alta variabilidade para a espécie / Abstract: Dermatobia hominis (Linnaus Jr.), the human bot fly, is one of the most important agent of primary myiases in vertebrates and one of the most important pests of the national cattle breeding and causes large losses in the industry of leather, milk and meat. The bot fly has an interesting biology and a very peculiar life cycle. These facts make difficult to observe the specie in nature and to breed it in laboratory. Due to these reasons, the characterization of the genetic variability and structure are important tools in the understanding of the D. hominis evolution. Techniques based on RAPD, with six populations, and RFLP of mitochondrial DNA, with twelve populations were applied for this purpose. Estimates of several biometrical procedures applied to the data were compared. Comparisons were also made with results of previous studies. Different approaches for analyzing RAPD data were also compared. The results of RAPD and mtDNA suggest that the specie behave as a metapopulation whose populations are variable and present gene flow among themselves. Results of different methods based on RAPD indicated that the Dice and Jaccard measures, calculated :from the complete matrix of data, and of Nei distances (1972) with the modifications of Lynch & Milligan (1994) were more adequate for the study of the variability of D. hominis. With the use of rntDNA RFLP, seventy-two haplotypes were detected, indicating high variability for this specie / Mestrado / Genetica Animal e Evolução / Mestre em Genética e Biologia Molecular

Orientador: Teresa Cristina Goulart de Oliveira Sequeira / Resumo: O presente trabalho foi desenvolvido com o objetivo de caracterizar os produtos de excreção/secreção (PE/S) de larvas de Dermatobia hominis, especialmente, no que se refere à atividade proteolítica desses produtos. Os PE/S foram obtidos de larvas de primeiro estágio (L1) cultivadas em laboratório e de larvas de segundo (L2) e terceiro (L3) estágios colhidas de bovinos parasitados. O perfil das proteínas foi obtido pela eletroforese dos PE/S em gel de poliacrilamida contendo sódio dodecil sulfato (SDS-PAGE) e a atividade proteolítica foi investigada utilizando gelatina, azocaseína e N-a-benzoil-arginina-nitroanilida (BAPNA) como substratos. Para caracterização das proteases foram realizados ensaios utilizando estes mesmos substratos, nos quais as amostras foram tratadas com os inibidores: PMSF, TPCK, TLCK, DCI, E-64, EDTA, Elastatinal, Leupeptina, Fenantrolina e Antipaína. Nos PE/S de L1 foram detectadas exclusivamente proteases com peso molecular aparente acima de 168 kDa, cuja inibição revelou pertencerem aos grupos das metalo-proteases e serina-proteases. Os zimogramas dos PE/S de L2 e L3 em géis copolimerizados com gelatina revelaram uma ampla faixa de atividade proteolítica, na qual estavam presentes proteases de alto, médio e baixo pesos moleculares aparentes. Nos ensaios realizados com inibidores de proteases, utilizando os três substratos, as proteases presentes em L2 e L3 foram identificadas como pertencentes ao grupo das serina-proteases, sendo em L3, predominantemente, do tipo tripsina. As alterações nos padrões de proteólise e nas características bioquímicas das proteases presentes nos três estágios larvais discuti- se à atividade das larvas em cada fase do seu desenvolvimento. / Abstract: In the present investigation the biochemical characteristics of the Dermatobia hominis larvae secretory/excretory products (PE/S) were analyzed mainly regard to their proteolytic activities. The PE/S of first-instar larvae were collected from larvae reared in the laboratory and of the second and third instars from larvae removed from infested cattle.The PE/S were tested in a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to identify their proteic profiles and the proteases activity were investigated using gelatine, azocasein, and N-a-benzoil-arginine-nitroanilide (BAPNA) as substrates. The proteases characterizations were performed by treating PE/S samples by the following proteases inhibitors: PMSF, TPCK, DCI, E-64, EDTA, Elastatinal, Leupeptine, Phenatroline, and Antipain. SDS-PAGE-Gelatin profiles of L1 PE/S revealed only proteases with molecular weight above 168 KDa whereas profiles from L2 and L3 PE/S revealed a high range of proteolytic activity, including high, medium and low molecular weight proteases. The assays performed with the protease inhibitors, and gelatin and azocasein as substrates, revealed that metalloprotease is the major class of proteases present in the PE/S of L1 besides the low content of serine-proteases. The major class of proteases present in the PE/S of L2 and L3 was characterized as serine-proteases, being in L3 predominantly of trypsin type. The changes in the proteolytic patterns and biochemical characteristics of the proteases found in all three instar larvae of D. hominis were with the larval activity in each phase of their development. / Mestre

Made available in DSpace on 2016-04-28T20:16:27Z (GMT). No. of bitstreams: 1 2008 - Fabio Silva de Souza.pdf: 3760278 bytes, checksum: d38f28c39076f3f01e4393fd0f31bc10 (MD5) Previous issue date: 2008-11-26 / The aim of this study was to characterize the association between the seasonality of dermatobiosis in bovines and its dipterans potential vectors as well as evaluate the association among the counts of Dermatobia hominis larvae and of potential vectors with meteorologicals data. It also had the objective of delimiting, quantifying and characterizing the geographical space of Serop?dica and Itagua? municipalities in relation to the favorable degrees for the occurrence of dermatobiosis in bovines in the rainy and dry periods using the Vista SAGA? 2007 information system. The seasonal fluctuation of D. hominis larvae was obtained by counts in ten bovines and of muscoids dipterans by capture using Adultrap? traps with sardine bait, once a month each, in three properties named A, B and C, localized in the Serop?dica, Paracambi and Itagua? municipalities, respectively, in the period of January 2006 to December 2007. The meteorological data of compensated mean temperature, pluvial precipitation and relative humidity were obtained in the Agricultural Ecology Station (22?48'S, 43?41'W and 33m of altitude). The associations between D. hominis larvae fluctuations, adult dipterans and climatological data were verified using the Spearman correlation test. Kruskal-Wallis and Mann-Whitney tests were used in the analysis of the significant differences between the rainy and dry periods in each property in the monthly average of D. hominis larvae, in the total monthly number and of dipterans family and in the analysis of other studies. In the elaboration of models by geoprocessing, some abiotic factors were used: occupation and covering of the land, altitude, declivity, soils and climatic factors, in which the evaluation, signature and monitorship functions of the Vista SAGA? 2007 program were applied. The infestation by D. hominis larvae in bovines was present during the 24 months of study in the three properties. A total of 18.966 dipterans potential vectors was captured, being 30,16% in the A property, 34,59% in the B property and 35,25% in the C property but there were no significant statistic difference between these. There was a positive and significant correlation (rs=0,63) between monthly averages of D. hominis larvae and relative humidity in 2006 at the A property; there was a positive and significant correlation between the total number of captured dipterans and pluvial precipitation (rs=0,80) and temperature (rs=0,60) at the B and C properties, respectively, in 2007. It was also observed a positive and significant correlation between the total number of captured dipterans of the Sarcophagidae family (rs=0,60) with the temperature in the C property in 2006, and between Muscidae (rs=0,67) and Calliphoridae (rs=0,76) with pluvial precipitation in the B property in 2007. There wasn t an association between monthly averages of D. hominis larvae and dipterans. It was verified a pattern lack in the association between monthly and periods fluctuations, rainy and dry of D. hominis larvae with the dipterans potential vectors fluctuations and of both with meteorologicals data. The geoprocessing allowed delimiting, quantifying and characterizing the potential of space temporal distribution of dermatobiosis in bovines. / Este estudo teve como objetivos caracterizar a associa??o entre a sazonalidade da dermatobiose em bovinos e seus d?pteros potenciais vetores, assim como avaliar a associa??o entre as contagens de larvas de Dermatobia hominis e de potenciais vetores com os dados meteorol?gicos. Teve tamb?m como objetivo, delimitar, quantificar e caracterizar o espa?o geogr?fico dos munic?pios de Serop?dica e Itagua? quanto aos gradientes de favorabilidade para a ocorr?ncia da dermatobiose em bovinos nos per?odos chuvoso e seco utilizando o sistema de informa??o Vista SAGA? 2007. A flutua??o sazonal de larvas de D. hominis foi obtida por contagens em dez bovinos e a de d?pteros musc?ides por captura, usando armadilhas Adultrap? com isca de sardinha, ambos uma vez ao m?s, nas tr?s propriedades denominadas A, B e C, localizadas nos munic?pios de Serop?dica, Paracambi e Itagua?, respectivamente, no per?odo de janeiro de 2006 a dezembro de 2007. Os dados meteorol?gicos de temperatura m?dia compensada, precipita??o pluvial e umidade relativa, foram obtidos na Esta??o Ecologia Agr?cola (22?48'S, 43?41'W e 33m de altitude). As associa??es entre as flutua??es de larvas de D. hominis, d?pteros adultos e dados climatol?gicos foram verificadas utilizando-se o teste de correla??o de Spearman. Os testes de Kruskal-Wallis e Mann-Whitney foram empregados na an?lise das diferen?as significativas entre os per?odos, chuvoso e seco, por propriedade, para a m?dia mensal de larvas de D. hominis, o n?mero mensal total e por fam?lia de d?pteros e na an?lise de dados publicados em outros estudos. Para a elabora??o dos modelos por geoprocessamento, utilizaram-se os fatores abi?ticos: uso e cobertura das terras, altitude, declividade, solos e fatores clim?ticos, aplicando-se as fun??es avalia??o, assinatura e monitoria do programa Vista SAGA? 2007. A infesta??o por larvas de D. hominis em bovinos esteve presente ao longo dos 24 meses de estudo nas tr?s propriedades. Um total de 18.966 d?pteros potenciais vetores foi capturado. Deste, 30,16% foram capturados na propriedade A, 34,59% na propriedade B e 35,25% na propriedade C, mas sem diferen?a estat?stica significativa entre estas. Houve correla??o positiva e significativa (rs=0,63) entre as m?dias mensais de larvas de D. hominis e umidade relativa, no ano de 2006, na propriedade A; houve correla??o positiva e significativa entre o n?mero total de d?pteros capturados e precipita??o pluvial (rs=0,80) e tamb?m com a temperatura (rs=0,60) nas propriedades B e C, respectivamente, no ano de 2007. Observou-se tamb?m correla??o positiva e significativa entre o n?mero total de d?pteros capturados da fam?lia Sarcophagidae (rs=0,60) com a temperatura na propriedade C no ano de 2006 e entre Muscidae (rs=0,67) e Calliphoridae (rs=0,76) com precipita??o pluvial na propriedade B em 2007. N?o ocorreu associa??o entre as m?dias mensais de larvas de D. hominis e de d?pteros. Constatou-se uma falta de padr?o na associa??o entre as flutua??es mensais e por per?odos, chuvoso e seco, das larvas de D. hominis com as flutua??es de d?pteros potenciais vetores e de ambas com os dados meteorol?gicos. O geoprocessamento permitiu delimitar, quantificar e caracterizar o potencial de distribui??o espa?o-temporal da dermatobiose nos per?odos chuvoso e seco.

Resumo: Avaliou-se comparativamente duas formulações, de ação prolongada, contendo Ivermectina (4% e 3,15%) em quatro experimentos em bovinos. Pelo grupo controle, constatou-se a presença das seguintes espécies de nematódeos gastrintestinais: Haemonchus placei, Cooperia punctata, Cooperia spatulata, Trichostrongylus axei, Oesophagostomum radiatum e Trichuris discolor. Quanto à eficácia anti-helmíntica, observou-se que a Ivermectina 4%* foi estatisticamente (P<0,05) superior à Ivermectina 3,15% contra Haemonchus placei, Cooperia punctata e Oesophagostomum radiatum. As duas formulações praticamente não diferiram quanto à eficácia anti-Boophilus microplus e antilarvas de Dermatobia hominis (berne). Decorridos 120 dias pós-tratamento, foram registrados, em relação ao grupo controle, diferenciais de ganho de peso corporal de 17,79 e 11,19 kg em bovinos medicados com Ivermectina 4% e Ivermectina 3,15%, respectivamente. / Abstract: Comparative endectocide efficacy and effect on weight gain in cattle treated with two (3.15% or 4%) ivermectin formulations. Four trials were conduced to a comparative evaluation of two long action (3.15% or 4%) ivermectin formulations. In the anthelmintic study, the 4% ivermectin formulation showed efficacy statistically higher than 3.15% ivermectin against Haemonchus placei, Cooperia punctata and Oesophagostomum radiatum. Ivermectin at both concentrations was similarly effective against Boophilus microplus and larvae of Dermatobia hominis (warble fly). After 120 days post-treatment, it was observed, relative to the control group, difference in the weight gain of 17.70kg and 11.10kg in cattle treated with 4% ivermectin and 3.15% ivermectin, respectively. / Orientador: Gilson Pereira de Oliveira / Coorientador: Alvimar José da Costa / Banca: Márcia Cristina de Sena Oliveira / Banca: Oeliton Ferreira Barbosa / Mestre

Made available in DSpace on 2016-04-28T20:16:24Z (GMT). No. of bitstreams: 1 2007- Fabiano Araujo Pires-01.pdf: 878766 bytes, checksum: 9179bfb28ddd6cee14d8d4414534b8f8 (MD5) Previous issue date: 2007-04-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / We performed a combination of proteinase assay, either in solution or immobilized in sodium dodecyl sulfate-polyacrylamide gel copolymerized with gelatin, to detect and quantify proteinases of Dermatobia hominis second (L2) and third (L3) instar larvae. In quantitative assays, we examined proteinase activity by hydrolysis of a panel of peptide bonds specific for the main proteinase classes. We verified that the pGlu-Phe-Leu p-nitroanilide substrate was hydrolyzed by crude extracts of L2 (3.0 ? 0.2 nmoles hour-1 mg of protein-1) and L3 (7.7 ? 0.1 nmoles hour-1 mg of protein-1) and that both activities were partially inhibited by transepoxysuccinyl- L-leucylamido-(4-guanidino)butane, 15 % and 3 % respectively. Also, we demonstrated that the Na-p-Tosyl-L-Arg methyl ester substrate was hydrolyzed by crude extracts of L2 (117 ? 24 nmoles hour-1 mg of protein-1) and L3 (111 ? 10 nmoles hour-1 mg of protein-1), suggesting a predominance of esterase activity in the crude larval preparation. Interestingly, the specific activity of serine-proteinases was totally inhibited by Phenylmethylsulphonyl fluoride in the L3 crude extract, while only 10 % of this enzyme class activity was inhibited in the L2 crude extract. Also, we have detected crude extract L2 (Km = 7,59) larvae have more affinity than L3 larvae (Km = 35,75) to Na-p-Tosyl-L-Arg methyl ester. The results of the qualitative assays with substrate gels suggested that L2 and L3 larvae express serine-proteinases with similar (13 kDa and 22 kDa) and distinct (50 kDa in L2 and 30 kDa in L3) relative molecular masses. Additionally, we have isolated an enriched esterase activity from L3 crude extract using successive chromatographies in Aprotinine-Agarose and DEAE-Sephacell columns. By this strategy we detected only one 50 kDa proteinase in this larvae crude extract. Finally, these findings contribute to the biochemical characterization of D. hominis L2 and L3 larvae. / Neste trabalho foram realizados ensaios de atividade de proteinase em solu??o e com prote?nas imobilizadas em gel de poliacrilamida contendo dodecil sulfato de s?dio copolimerizado com gelatina, para detec??o e quantifica??o das proteinases presentes nos extratos larvares de segundo (L2) e terceiro (L3) est?gios de Dermatobia hominis. Nos ensaios quantitativos, utilizou-se um painel de pept?deos sint?ticos espec?ficos para as principais classes de proteinases. Verificamos que o substrato pGlu-Phe-Leu p-nitroanilide foi hidrolisado pelo extrato total de L2 (3,0 ? 0,2 nmoles hora-1 mg de prote?na-1) e L3 (7,7 ? 0,1 nmoles hora-1 mg de prote?na-1) e que ambas atividades foram parcialmente inibidas pelo trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane, 15 % e 3 % respectivamente. Tamb?m, demonstramos que o substrato Na-p-Tosyl-L-Arg methyl ester foi hidrolisado pelos extratos totais de L2 (117 ? 24 nmoles hora-1 mg de prote?na-1) e L3 (111 ? 10 nmoles hora-1 mg de proteina-1), sugerindo uma predomin?ncia da atividade ester?sica nestes extratos. A atividade espec?fica de serino-proteinases foi totalmente inibida pelo phenylmethylsulphonyl fluoride nos extratos de L3, enquanto que somente 10 % desta atividade foi inibida nos extrados de L2. Al?m disso, n?s detectamos que o extrato total das larvas L2 (Km = 7,59) tem maior afinidade ao Na-p-Tosyl-L-Arg methyl ester do que o extrato total das larva de L3 (Km = 35,75). Os resultados do ensaio qualitativo com g?is de substrato sugerem que os extratos larvares L2 e L3 expressam serino-proteinases com similares (13 kDa e 22 kDa) e distintas (50 kDa em L2 e 30 kDa em L3) massas moleculares relativas. Adicionalmente, isolamos uma atividade ester?sica enriquecida do extrato total de L3 utilizando sucessivas cromatografias em colunas de Aprotinina-Agarose e DEAE-Sephacell. Com esta estrat?gia, detectamos somente uma banda de proteinase de 50 kDa neste extrato total. Estes resultados contribuem para a caracteriza??o das proteinases larvares de D. hominis.

Made available in DSpace on 2014-06-11T19:27:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-06-29Bitstream added on 2014-06-13T20:57:21Z : No. of bitstreams: 1 canavaci_fht_me_jabo.pdf: 217331 bytes, checksum: b1d45ab310a2e588fc1485227ef1cd7d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Avaliou-se comparativamente duas formulações, de ação prolongada, contendo Ivermectina (4% e 3,15%) em quatro experimentos em bovinos. Pelo grupo controle, constatou-se a presença das seguintes espécies de nematódeos gastrintestinais: Haemonchus placei, Cooperia punctata, Cooperia spatulata, Trichostrongylus axei, Oesophagostomum radiatum e Trichuris discolor. Quanto à eficácia anti-helmíntica, observou-se que a Ivermectina 4%* foi estatisticamente (P<0,05) superior à Ivermectina 3,15% contra Haemonchus placei, Cooperia punctata e Oesophagostomum radiatum. As duas formulações praticamente não diferiram quanto à eficácia anti-Boophilus microplus e antilarvas de Dermatobia hominis (berne). Decorridos 120 dias pós-tratamento, foram registrados, em relação ao grupo controle, diferenciais de ganho de peso corporal de 17,79 e 11,19 kg em bovinos medicados com Ivermectina 4% e Ivermectina 3,15%, respectivamente. / Comparative endectocide efficacy and effect on weight gain in cattle treated with two (3.15% or 4%) ivermectin formulations. Four trials were conduced to a comparative evaluation of two long action (3.15% or 4%) ivermectin formulations. In the anthelmintic study, the 4% ivermectin formulation showed efficacy statistically higher than 3.15% ivermectin against Haemonchus placei, Cooperia punctata and Oesophagostomum radiatum. Ivermectin at both concentrations was similarly effective against Boophilus microplus and larvae of Dermatobia hominis (warble fly). After 120 days post-treatment, it was observed, relative to the control group, difference in the weight gain of 17.70kg and 11.10kg in cattle treated with 4% ivermectin and 3.15% ivermectin, respectively.

Made available in DSpace on 2014-06-11T19:24:00Z (GMT). No. of bitstreams: 0 Previous issue date: 2004Bitstream added on 2014-06-13T19:10:01Z : No. of bitstreams: 1 brant_mprc_me_botfmvz.pdf: 347841 bytes, checksum: 07a65a6f458afcc54be76d10fc51457e (MD5) / Universidade Estadual Paulista (UNESP) / O presente trabalho foi desenvolvido com o objetivo de caracterizar os produtos de excreção/secreção (PE/S) de larvas de Dermatobia hominis, especialmente, no que se refere à atividade proteolítica desses produtos. Os PE/S foram obtidos de larvas de primeiro estágio (L1) cultivadas em laboratório e de larvas de segundo (L2) e terceiro (L3) estágios colhidas de bovinos parasitados. O perfil das proteínas foi obtido pela eletroforese dos PE/S em gel de poliacrilamida contendo sódio dodecil sulfato (SDS-PAGE) e a atividade proteolítica foi investigada utilizando gelatina, azocaseína e N-a-benzoil-arginina-nitroanilida (BAPNA) como substratos. Para caracterização das proteases foram realizados ensaios utilizando estes mesmos substratos, nos quais as amostras foram tratadas com os inibidores: PMSF, TPCK, TLCK, DCI, E-64, EDTA, Elastatinal, Leupeptina, Fenantrolina e Antipaína. Nos PE/S de L1 foram detectadas exclusivamente proteases com peso molecular aparente acima de 168 kDa, cuja inibição revelou pertencerem aos grupos das metalo-proteases e serina-proteases. Os zimogramas dos PE/S de L2 e L3 em géis copolimerizados com gelatina revelaram uma ampla faixa de atividade proteolítica, na qual estavam presentes proteases de alto, médio e baixo pesos moleculares aparentes. Nos ensaios realizados com inibidores de proteases, utilizando os três substratos, as proteases presentes em L2 e L3 foram identificadas como pertencentes ao grupo das serina-proteases, sendo em L3, predominantemente, do tipo tripsina. As alterações nos padrões de proteólise e nas características bioquímicas das proteases presentes nos três estágios larvais discuti- se à atividade das larvas em cada fase do seu desenvolvimento. / In the present investigation the biochemical characteristics of the Dermatobia hominis larvae secretory/excretory products (PE/S) were analyzed mainly regard to their proteolytic activities. The PE/S of first-instar larvae were collected from larvae reared in the laboratory and of the second and third instars from larvae removed from infested cattle.The PE/S were tested in a sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) to identify their proteic profiles and the proteases activity were investigated using gelatine, azocasein, and N-a-benzoil-arginine-nitroanilide (BAPNA) as substrates. The proteases characterizations were performed by treating PE/S samples by the following proteases inhibitors: PMSF, TPCK, DCI, E-64, EDTA, Elastatinal, Leupeptine, Phenatroline, and Antipain. SDS-PAGE-Gelatin profiles of L1 PE/S revealed only proteases with molecular weight above 168 KDa whereas profiles from L2 and L3 PE/S revealed a high range of proteolytic activity, including high, medium and low molecular weight proteases. The assays performed with the protease inhibitors, and gelatin and azocasein as substrates, revealed that metalloprotease is the major class of proteases present in the PE/S of L1 besides the low content of serine-proteases. The major class of proteases present in the PE/S of L2 and L3 was characterized as serine-proteases, being in L3 predominantly of trypsin type. The changes in the proteolytic patterns and biochemical characteristics of the proteases found in all three instar larvae of D. hominis were with the larval activity in each phase of their development.

Made available in DSpace on 2016-04-28T20:15:30Z (GMT). No. of bitstreams: 1 2008- Joice Aparecida Rezende Vilela.pdf: 1244449 bytes, checksum: 335f88e6cd41b29936d7ce97d585c143 (MD5) Previous issue date: 2008-03-04 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Dermatobia hominis, the human botfly, has obligatory parasitic larval forms of the subcutaneous tissue of domestic animals, wildlife and man, causing nodule myiasis. Its importance for the cattle industry is related to the economic damages caused by those larvae. The conventional chemical control has facing some problems such as the accelerated development of resistance and residues in animal products. The objective of this study was to evaluate the prevention of botfly myiasis through the use of phytotherapic neen and of the moxidectin observing the capacity of interference in the evolution of parasite biology. For later infestation, third instar larvae were collected and kept in BOD at temperature of 27 ? 1?C and 10% R.U for pupation. After 24 to 31 days, emerged adults were placed in cages with Musca domestica and Chrysomya albiceps, used as a vector for oviposition of D. hominis. The vector carrying eggs were captured and placed in BOD. After 4 to 6 days, first instar larvae were maintained in BOD at 19? C, until host infestation. In the experimental lineation was used 12 male cattle free of D. hominis natural infestation. The animals were previously placed in screened stalls in the W.O. NEITZ Research Station at UFRRJ, receiving concentrated feed, grass, and water. The cattle was distributed into three groups with four animals and submitted to the treatments, being a control animal (treated with water pouredon), two animals treated with commercial products A and B of neem oil (Azadirachta indica, 2000 ppm of the active principle Azadirachtin) applied as a pour-on (50 ml for animal), and one animal treated with moxidectin 10% long action injected subcutaneously (1ml/100 kg) in the back of the ear. Each animal was infested with 30 first instar larvae (L1) on days 03, 07, 14, and 21 after treatment. The outbreak of L1 was stimulated by thermal source and larvae were transferred with a fine brush to a shaved region along the bovine back. The infestations were mapped for monitoring every two days to evaluation of efficacy and residual effect of treatments in accordance with the larval survival at different periods of infestation. With the purpose of verify possible collateral effects, were accomplished clinical and laboratorial examinations of the animals, before and 15 days after the treatments. Regarding efficacy, the neem products applied as pour-on had statistical not significant effectiveness for inhibition of larvae development. On the other hand, injectable moxidectin 10% showed 100% efficacy until day 14 and although larvae penetration was not precluded, larvae development was inhibited, but from day 21, did not inhibit the development of the penetrated larvae. The third instar larvae that developed after the treatments were collected and kept in BOD for observation of some biological parameters. It was observed that larvae from all treatments showed regular pupation, emergence of morphologically normal flies and the presence of viable postures. / Dermatobia hominis, a mosca do berne, possui formas larvais parasitas obrigat?rios do tecido subcut?neo de animais dom?sticos, silvestres e do homem, provocando mi?ase nodular. Sua import?ncia na bovinocultura relaciona-se aos preju?zos econ?micos determinados pelas formas evolutivas larvais. No controle qu?mico convencional tem sido observado desenvolvimento acelerado de resist?ncia e res?duos nos produtos animais. Os objetivos do experimento foram avaliar a preven??o da dermatobiose atrav?s do uso do fitoter?pico nim e da moxidectina, observando a capacidade de interfer?ncia na evolu??o da biologia parasit?ria. Para a infesta??o, foram coletadas larvas de terceiro instar, que foram mantidas em estufa B.O.D. ? temperatura de 27 ? 1?C e 70 ? 10% U.R para a pupa??o. Ap?s 24 a 31 dias, os adultos que emergiram foram colocados em gaiolas com Musca domestica e Chrysomya albiceps, vetores para oviposi??o de D. hominis. Os vetores portadores de ovos foram capturados e acondicionados em estufa B.O.D. Ap?s 4 a 6 dias, as larvas de primeiro instar, em eclos?o foram mantidas a 19?C, at? o momento da infesta??o. No delineamento experimental utilizaram-se 12 bovinos machos, livres de infesta??o por D. hominis. Os animais foram estabulados em baias teladas na Esta??o de Pesquisas Parasitol?gicas W.O. NEITZ, da UFRRJ, recebendo ra??o concentrada, capim picado e ?gua ad libitum. Os bovinos foram distribuidos em tr?s grupos com quatro animais e submetidos aos tratamentos, sendo um animal controle (tratado com ?gua aplicada pour-on), dois animais tratados com produtos comerciais A e B ? base de ?leo de nim (Azadirachta indica, concentra??o de 2000 ppm do princ?pio ativo Azadirachtina) em aplica??o pour-on ao longo do dorso dos animais, dose de 50 ml por animal, e um tratado com moxidectina ? 10% Longa A??o, dose de 1ml/100 kg, em inje??o subcut?nea na parte posterior da orelha. Cada animal foi infestado com 30 larvas L1 nos dias 03, 07, 14 e 21 ap?s os tratamentos. A eclos?o das L1 foi estimulada por fonte t?rmica, e as mesmas colocadas com um pincel fino sobre a regi?o tricotomizada ao longo do dorso. As infesta??es foram mapeadas para acompanhamento a cada dois dias para avalia??o da efic?cia e tempo residual dos tratamentos de acordo com a sobreviv?ncia larval nos diferentes per?odos de infesta??o. Com a finalidade de se verificar poss?veis efeitos colaterais foram realizados exames cl?nico e laboratorial dos animais, antes e 15 dias ap?s os tratamentos. N?o foram observados altera??o dos par?metros cl?nico-laboratoriais. Com rela??o ? efic?cia, os produtos do Nim tiveram efic?cia estatisticamente n?o significativa na forma de aplica??o pour-on, na inibi??o do desenvolvimento parasit?rio. A moxidectina ? 10% apresentou efic?cia de 100% at? o 14? dia, n?o impedindo a penetra??o da larva mas a evolu??o parasit?ria, e a partir do 21? dia n?o inibiu o desenvolvimento das larvas penetradas. As larvas L3 que se desenvolveram ap?s os tratamentos, foram coletadas e mantidas em B.O.D para observa??o de alguns par?metros biol?gicos. Constatou-se que as larvas das L3 provenientes de todos tratamentos evoluiram para pupa??o, emerg?ncia de moscas morfologicamente normais e presen?a de posturas vi?veis.

La lechiguana es una enfermedad de los bovinos definida como una paniculitis fibrogranulomatosa proliferativa. Se caracteriza por una tumoración subcutánea de crecimiento rápido, con marcada proliferación de tejido fibroso que puede ocasional la muerte del animal. El objetivo de este trabajo fue confirmar la hipótesis de que la lechiguana es una enfermedad causada por una asociación entre el parasitismo de la Dermatobia hominis y la presencia de Mannheimia granulomatis.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-06-13T13:31:56Z No. of bitstreams: 1 2015 - William Soares Medeiros.pdf: 2112015 bytes, checksum: 79b821090426baeb106b29b980b2ad18 (MD5) / Made available in DSpace on 2017-06-13T13:31:56Z (GMT). No. of bitstreams: 1 2015 - William Soares Medeiros.pdf: 2112015 bytes, checksum: 79b821090426baeb106b29b980b2ad18 (MD5) Previous issue date: 2015-07-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Dermatobia hominis (Linneaus Jr., 1781) (Diptera, Cuterebridae), popularly known in English as the Human Botfly or Warble fly, the only species of this genus and endemic to the neotropical region, is a holometabolous insect with an independent phase of life in its adult form and an obligatory parasitic phase in its larval form. In its parasitic phase, the larva is the cause of furuncular myiasis occurring in a large variety of endothermic animals, with most frequent occurrence in cattle, dogs and humans. The lack of information concerning dermatobiosis creates difficulties in its diagnosis. By means of a systematic review of the literature on the subject, it can be verified that this fly shows versatility in its phoresis and has a vast list of phoretic records. Field studies are restricted to observation of its larval form, because its adult form is difficult to observe, making this a rare event. With the intention of evaluating and comparing fluctuation in the population of D. hominis, searching for correlations between the incidence of Botfly larval infestations and climatic factors and their distribution on the body surface of cattle, two areas of study were selected, in which field collections were carried out twice monthly for counting the number of larvae in the herds. Discrepancies were discovered in the average number of larval nodes in cattle between these areas of study. These nodules showed variation in their bodily distribution on the cattle, with a higher incidence in the anterior region, compromising the quality of the animals? skins for economic purposes. In one of the areas, there was a correlation between the average number of larvae and the temperature and precipitation. These climatic factors are related to edaphic characteristics, influencing the penetration of the L3 larvae and the pupal period. For a greater understanding of the dynamics and fluctuations in the population of D. hominis in these areas, ample monitoring is needed to determine if the results encountered in this study are atypical or influenced by local characteristics. Currently, the demand for research is for methods of controlling the dermatobiosis, with emphasis on integrated plague management. Other deficiencies include following the Botfly?s complete life cycle in the laboratory and understanding certain peculiar adaptations, such as atrophied oral apparatus / A Dermatobia hominis (Linneaus Jr., 1781) (Diptera, Cuterebridae), popularmente conhecida como mosca do berne, ?nica esp?cie do g?nero e end?mica da regi?o Neotropical, ? um inseto holomet?bolo com uma fase de vida livre na sua forma adulta e uma fase parasit?ria obrigat?ria em sua forma larval. Em sua fase parasit?ria, a larva ? causadora da mi?ase furuncular ocorrendo em uma grande variedade de animais endot?rmicos, com ocorr?ncia mais frequente em bovinos, c?es e no homem. A falta de informa??o sobre a dermatobiose gera dificuldade em seu diagn?stico. Atrav?s de uma revis?o sistem?tica da literatura sobre o assunto, verifica-se que a mosca apresenta versatilidade para a foresia e conta com vasta lista de registros de for?ticos. Os estudos em campo se restringem a observa??es sobre sua forma larval, pois sua forma adulta ? de dif?cil observa??o tornando este evento raro. Visando avaliar e comparar a frequ?ncia populacional de D. hominis, buscando correla??o entre a incid?ncia do berne e fatores clim?ticos e a sua distribui??o na superf?cie corporal de bovinos, foram selecionadas duas ?reas de estudo, as quais foram realizadas coleta de campo quinzenalmente para o levantamento do n?mero de bernes no rebanho. Foram encontradas diverg?ncias no n?mero m?dio de n?dulos de berne em bovinos entre as ?reas de estudo. Os n?dulos apresentaram varia??o na distribui??o corporal dos bovinos, tendo maior incid?ncia na regi?o anterior, comprometendo a qualidade da pele do animal para fins econ?micos. Em uma das ?reas houve correla??o entre o n?mero m?dio de bernes e a temperatura e precipita??o. Esses fatores clim?ticos est?o relacionados ?s caracter?sticas ed?ficas, influenciando a penetra??o das larvas L3 e o per?odo pupal. Para o maior entendimento da din?mica e frequ?ncia populacional da D. hominis nessas ?reas, h? necessidade de amplo monitoramento para averiguar se os resultados encontrados neste estudo s?o at?picos ou influenciados por caracter?sticas locais. Atualmente a demanda de pesquisa est? em m?todos para controle da dermatobiose, com destaque para o manejo integrado de pragas. Outras lacunas est?o na realiza??o de seu ciclo completo em laborat?rio e compreens?o de adapta??es peculiares, tais como aparelho bucal atrofiado.