Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-10-18T12:35:34Z No. of bitstreams: 1 2016 - Tatiana Pires Pereira.pdf: 2546753 bytes, checksum: c1673b0ae2c6812a3f60faa79b5fae75 (MD5) / Made available in DSpace on 2017-10-18T12:35:34Z (GMT). No. of bitstreams: 1 2016 - Tatiana Pires Pereira.pdf: 2546753 bytes, checksum: c1673b0ae2c6812a3f60faa79b5fae75 (MD5) Previous issue date: 2016-07-27 / Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do RJ - FAPERJ / This work was divided in four chapters, in which the first was performed with the goal of quantifying the condensed tannin (CT) content by the Stiasny?s reaction and to determine the classes of secondary metabolites present by the phytochemical prospection technique and magnetic resonance in the tropical forage legumes Cajanus cajan (guandu - GUA), Gliricidia sepium (gliricidia - GLI), Flemingia macrophylla (flemingia - FLE), Cratylia arg?ntea (cratilia - CRA), Mimosa caesalpineafolia (sabi?) (this legume divided into bark and leaf fraction ? SABc and SABf) among the treatments. The extracts obtained were divided into: total extract, number of Stiasny (NS), CT and non-tannins. The FLE, CRA, GUA,GLI, SABf and SABc had obtained the following values for total extract: 13.20; 13.06; 8.28; 14.73; 15.67 and 6.22%, respectively. The reactivity by NS, in the same order of legumes, was 11.25; 4.54; 7.37; 6.70; 23.06 and 71.62%, whereas the CT presented the following values: 1.52; 0.59; 0.61; 0.96; 3.6 and 4.43%, and non-tannin was 11.68; 12.46; 7.67; 13.75; 12.07 and 1.76%, respectively. The following classes of secondary metabolites were identified with greater evidence: saccharides, carbohydrates, non-protein amino acids and glicos?deos cardioativos. For the CT, the intensity was low for most of the legumes, with greater content in CRA, GUA and SABf. The wain compound in the extracts was methyl-inositol (sugar). The second chapter had the objective of assaying in the legumes mentioned above and one more specie, Stylosanthes spp. (estilosantes-EST), condensed tannin (CT) constituents, with the use of organic solvents, soluble CT (ECT), CT adhered to protein (PBCT), CT adhered to fiber (FBCT), and total CT (TCT), CT structural pro-pelargonidin (PP); prodelfinidin (PD) and procyanidin (PC), molecular weight (polymerization degree (DP), molecular distance distributed of the polymer (PDI); average weight of molecular mass (Mw), and average number of molecular mass (Mn), and the biological activity through precipitated proteins by phenols (PPP). The variables ECT, PBCT, and TCT presented were influenced by different species (P?0.05). The FBCT fraction was not found in the legumes. Molecular weights (DP, PDI, Mw e Mn) were affected by the different species (P?0.05), ranging from 737 to 1168 da. The structural characteristics (PP, PD, PC and PD:PC) varied among the species. In the third chapter I evaluated methanogenesis (total methane (CH4total)), incubated (CH4inc) and fermented (CH4ferm) and ruminal fermentation parameters total gas production (PGT), pH, ammonium (N-NH3), short-chain fatty acids (SCFA) and in vitro organic matter disappearance (IVOMD) as they related to CT present in the legumes and Urochloa brizantha cv. marandu hay as control (CTL). The effect of polyethylene glycol (PEG) on the leaf fraction of sabi? was tested as well, which had a CT content of 15.97%. No alteration in the pH (P?0.05) for the treatments evaluated. However, a decrease of total gas and methane production for all the treatments with presence of CT (P?0.05). When PEG was added, there was a 27.01 (8% PEG) and 35.01 (16% PEG) increase in total gas production and 3.59 (8% PEG) and 4.15 (16% PEG) of methane production. GUA, FLE, SABc and SABf were capable of modifying (P?0.05) the content of NH3-N (mg/dL), along with the CTL, which also presented lower values compared to legumes with no or only traces of CT (ETL, CRA and GLI). There was significant difference (P?0.05) for IVOMD between the legumes and control, it was observed lower disappearance (P?0.05) for FLE, GUA and SABf in relation to CTL, while SABc did disappear. The SABf IVOMD was affected by the addiction of PEG. There was lower digestibility for FLE, GUA and SABf in relation to the CTL, while the bark fraction of SAB did not disappear at all. On the SCFA profile, there was difference (P?0.05) among the treatments evaluated, with lower values for the legumes with presence of CT. In the fourth chapter I tested the effect of CT from the legumes in study (FLE, CRA, GUA, GLI, EST, SABf and SABc) on larval migration inhibition (LMI) in vitro, on the infective larvae L3 of the nematode Haemonchus contortus (HC), compared with Ivermectin and a negative control (rumen fluid and buffer). Among the legumes studied, SABf and GUA did not differ (P?0.05), with the greater (P?0.05) LMI percentage (34.75% and 34.33%) than the other entries. The legumes GUA, FLE and SABc did not differ (P?0.05), presenting moderate values of LMI (30.25%, 30.0% and 29.75%, respectively). Among the legumes studied, the lowest LMI percentage was CRA (18.46%), GLI (23.75%) and negative control (rumen fluid and buffer), with values near (P?0.05) from to Ivermectin (22.0%). / Este trabalho foi dividido em quatro cap?tulos. O primeiro realizou-se com o objetivo de quantificar o teor de tanino condensado (TC) atrav?s da Rea??o de Stiasny e conhecer as classes de metab?litos secund?rios presentes pela t?cnica de prospec??o fitoqu?mica e resson?ncia magn?tica nas leguminosas forrageiras tropicais Cajanus cajan (guandu-GUA), Gliricidia sepium (gliricidia-GLI), Flemingia macrophylla (flemingia-FLE), Cratylia arg?ntea (cratilia-CRA), Mimosa caesalpineafolia (sabi?) sendo que essa leguminosa tinha a fra??o casca e folha (SABc e SABf) entre os tratamentos. Os extratos obtidos foram divididos em: extrato total, n?mero de Stiasny (NS), TC e n?o taninos. A FLE, CRA, GUA, GLI, SABf e SABc apresentaram os valores para o extrato total 13,20; 13,06; 8,28; 14,73; 15,67 e 6,22%, respectivamente. A reatividade pelo NS, na mesma ordem das leguminosas, foi de 11,25; 4,54; 7,37; 6,70; 23,06 e 71,62%, j? o TC apresentou os seguintes valores 1,52; 0,59; 0,61; 0,96; 3,6 e 4,43% e o n?o tanino foi de 11,68; 12,46; 7,67; 13,75; 12,07 e 1,76%, respectivamente. Foram identificadas as seguintes classes de compostos secund?rios em maiores evid?ncias: os sacar?deos, carboidratos, amino?cidos n?o prot?icos e os glicos?deos cardioativos. J? para o TC, a intensidade foi baixa para grande parte das leguminosas, prevalecendo maior teor para CRA, GUA e SABf. Foi constatado como componente principal nos extratos o metil-inositol (a??car). O segundo cap?tulo teve como objetivo avaliar nas leguminosas citadas acima e mais uma esp?cie, o Stylosanthes spp (estilosantes-EST), analisar os constituintes do TC com uso de solvente org?nico, tanino sol?vel (TCE), tanino aderido ? prote?na (TCPB), tanino aderido ? fibra (TCFB) e taninos condensados totais (TCT), caracter?sticas estruturais tais como: propelargonidina (PP); prodelfinidina (PD) e procianidina (PC); peso molecular (grau de polimeriza??o?(DP); dist?ncia do peso molecular distribu?do do pol?mero (PDI); peso m?dio da massa molecular (Mw); n?mero m?dio da massa molecular (Mn); al?m de determinar a atividade biol?gica, atrav?s da t?cnica de prote?nas precipit?veis por fen?is (PPP). As vari?veis TCE, TCPB e TCT apresentadas foram influenciadas pelas diferentes esp?cies (P?0,05). A fra??o TCFB n?o foi constatada nas leguminosas. Os pesos moleculares (Mw) foram influenciados pelas diferentes esp?cies (P?0,05), variando de 737 a 1168 Da. As caracter?sticas estruturais (PP, PD, PC e PD:PC) tiveram varia??o entre as esp?cies estudadas. Objetivou-se com o terceiro cap?tulo avaliar a metanog?nese (metano total (CH4-total), incubado (CH4 inc.) e fermentado (CH4 ferm.) e os par?metros de fermenta??o ruminal (produ??o de g?s total (PGT), pH, am?nia (N-NH3), ?cidos graxos de cadeia curta (AGCC) e digestibilidade in vitro da mat?ria org?nica (DIVMO) frente aos TC presentes nas leguminosas e feno de Urochloa brizantha cv. marandu como controle (CTL). Foi testado tamb?m o efeito do polietileno glicol (PEG) sobre a fra??o folha do sabi?, que teve conte?do de TC de 15,97%. N?o foi observado altera??o no pH (P?0,05) para os tratamentos avaliados. No entanto, foram observadas diminui??o da produ??o total de g?s e produ??o de metano para todos os tratamentos com presen?a de TC (P?0,05). Para o tratamento com PEG houve aumento de 27,01 (8% PEG) e 35,01 (16% PEG) na produ??o total de g?s e 3,59 (8% PEG) e 4,15 (16% PEG) na produ??o de metano. GUA, FLE, SABc e SABf foram capazes de modificar (P?0,05) a concentra??o de N-NH3 (mg/dL) juntamente com o CTL, que tamb?m apresentou valores inferiores comparado as leguminosas com tra?os e aus?ncia do TC (ETL, CRA e GLI). Houve diferen?a (P?0,05) para DIVMO entre as leguminosas e o controle, observou-se menor digestibilidade (P?0,05) para FLE, GUA e SABf, em rela??o ao CTL, n?o sendo digest?vel o SABc. A DIVMO foi afetada pela adi??o de PEG na dieta do SABf. No perfil dos AGCC houve diferen?a (P?0,05) para os tratamentos avaliados, com menor valor para as leguminosas com presen?a de TC. O quarto cap?tulo teve como objetivo testar o efeito da t?cnica de inibi??o da migra??o larval (IML) in vitro do TC proveniente das leguminosas em estudo (FLE, CRA, GUA, GLI, EST, SABf e SABc) sobre as larvas infectantes L3 do nemat?de o Haemonchus contortus (HC) comparando com Ivermectina e controle negativo (l?quido ruminal e tamp?o). Entre as leguminosas estudadas o SABf e GUA n?o diferiram entre si (P?0,05), com as maiores porcentagens IML (34,75% e 34,33%). As leguminosas GUA, FLE e SABc n?o diferiram entre si (P?0,05), apresentando moderados valores de IML (30,25%, 30,0% e 29,75%, respectivamente). Entre as leguminosas estudadas a menor porcentagem de IML foi para CRA (18,46%), GLI (23,75%) e controle negativo (l?quido de r?men e tamp?o) valores pr?ximos do controle positivo com Ivermectina (22,0%).