The objectives of this study were to investigate the effects of autoclave heating on the rumen protein degradation characteristics of flaxseed (<i>Linum usitatissimum</i>, cv. Vimy), and to compare them to differences in diffuse reflectance infrared Fourier transform (DRIFT) and Synchrotron based Fourier transform infrared microspectroscopy (S-FTIR) measurements of the protein alpha-helix to beta-sheet ratios. Hierarchical cluster analysis (CLA) and principal components analysis (PCA) were also conducted to identify differences in the DRIFT spectra. Flaxseed samples were kept raw for control or autoclaved in batches at 120°C for 20, 40 or 60 min for treatments 1, 2 and 3, respectively. The rumen degradation kinetics of protein were measured along with the protein sub-fractions of the Cornell net carbohydrate and protein system (CNCPS), and chemical composition. Intestinal digestibility was determined using the three-step procedure outlined by Calsamiglia and Stern (1995). Protein supply to the small intestine was determined using the NRC (2001) and DVE/OEB models. The results showed that heating increased dry matter (DM) and ether extract (EE) content, while reducing neutral detergent fibre (NDF) and acid detergent fibre (ADF), with little numerical difference between the three treatments. Soluble crude protein (SCP) also decreased upon autoclaving with concomitant increases in non-protein nitrogen (NPN), neutral detergent insoluble nitrogen (NDIN) and acid detergent insoluble nitrogen (ADIN). The CNCPS protein sub-fractions with the greatest changes were the buffer-soluble true protein fraction (PB1) and the fraction representing buffer-insoluble true protein which is not bound to NDF (PB2) showing dramatic increases, indicating a decrease in the overall protein degradability. <i>In situ</i>experiments showed a reduction in effective degradable dry matter (EDDM) as well as a reduction in effective degradable crude protein (EDCP) without significant differences between the treatments. Intestinal digestibility of protein as estimated by the three-step procedure showed no changes upon autoclaving. Modeling results, with flaxseed as the only feed source, for absorbable ruminally-undegraded feed protein in the intestines using both the NRC (2001) and DVE/OEB systems showed increases as a consequence of the autoclave treatments but again there were no differences between the treatments. The degraded protein balance results showed for both the NRC (2001) and DVE/OEB models that both were decreased upon autoclave treatment. However, the values for the NRC (2001) model suggested a potential nitrogen (N) deficiency and, therefore potentially impaired microbial crude protein (MCP) production, whereas the values for the DVE/OEB system showed potential N excess and, therefore, possible loss from the rumen. DRIFT analysis of protein secondary structure ratios showed a decrease in the alpha-helix to beta-sheet ratio for the whole seed, whereas results from S-FTIR spot data for cotyledon tissue showed autoclaving had the opposite effect on the ratio. CLA and PCA were successfully used to make distinctions between the different treatment spectra and showed enhanced sensitivity upon selection of a smaller spectral window to include only the amide I and II portion of the IR spectrum. The results failed to demonstrate any differences between the autoclave treatments used in this study, and showed that autoclaving generally decreased effectively ruminal degradability of flaxseed protein. The results further indicated that autoclaving had a significant enough effect on the flaxseed to permit identification of the altered alpha-helix to beta-sheet ratio with the mid-IR spectrum, as well as differentiation between the treatments using PCA and CLA. PCA and CLA results suggest that mid-IR spectral methods are more sensitive than traditional methods when used to identify differences between the heat treatments.

The objectives of this study were to investigate the effects of autoclave heating on the rumen protein degradation characteristics of flaxseed (<i>Linum usitatissimum</i>, cv. Vimy), and to compare them to differences in diffuse reflectance infrared Fourier transform (DRIFT) and Synchrotron based Fourier transform infrared microspectroscopy (S-FTIR) measurements of the protein alpha-helix to beta-sheet ratios. Hierarchical cluster analysis (CLA) and principal components analysis (PCA) were also conducted to identify differences in the DRIFT spectra. Flaxseed samples were kept raw for control or autoclaved in batches at 120°C for 20, 40 or 60 min for treatments 1, 2 and 3, respectively. The rumen degradation kinetics of protein were measured along with the protein sub-fractions of the Cornell net carbohydrate and protein system (CNCPS), and chemical composition. Intestinal digestibility was determined using the three-step procedure outlined by Calsamiglia and Stern (1995). Protein supply to the small intestine was determined using the NRC (2001) and DVE/OEB models. The results showed that heating increased dry matter (DM) and ether extract (EE) content, while reducing neutral detergent fibre (NDF) and acid detergent fibre (ADF), with little numerical difference between the three treatments. Soluble crude protein (SCP) also decreased upon autoclaving with concomitant increases in non-protein nitrogen (NPN), neutral detergent insoluble nitrogen (NDIN) and acid detergent insoluble nitrogen (ADIN). The CNCPS protein sub-fractions with the greatest changes were the buffer-soluble true protein fraction (PB1) and the fraction representing buffer-insoluble true protein which is not bound to NDF (PB2) showing dramatic increases, indicating a decrease in the overall protein degradability. <i>In situ</i>experiments showed a reduction in effective degradable dry matter (EDDM) as well as a reduction in effective degradable crude protein (EDCP) without significant differences between the treatments. Intestinal digestibility of protein as estimated by the three-step procedure showed no changes upon autoclaving. Modeling results, with flaxseed as the only feed source, for absorbable ruminally-undegraded feed protein in the intestines using both the NRC (2001) and DVE/OEB systems showed increases as a consequence of the autoclave treatments but again there were no differences between the treatments. The degraded protein balance results showed for both the NRC (2001) and DVE/OEB models that both were decreased upon autoclave treatment. However, the values for the NRC (2001) model suggested a potential nitrogen (N) deficiency and, therefore potentially impaired microbial crude protein (MCP) production, whereas the values for the DVE/OEB system showed potential N excess and, therefore, possible loss from the rumen. DRIFT analysis of protein secondary structure ratios showed a decrease in the alpha-helix to beta-sheet ratio for the whole seed, whereas results from S-FTIR spot data for cotyledon tissue showed autoclaving had the opposite effect on the ratio. CLA and PCA were successfully used to make distinctions between the different treatment spectra and showed enhanced sensitivity upon selection of a smaller spectral window to include only the amide I and II portion of the IR spectrum. The results failed to demonstrate any differences between the autoclave treatments used in this study, and showed that autoclaving generally decreased effectively ruminal degradability of flaxseed protein. The results further indicated that autoclaving had a significant enough effect on the flaxseed to permit identification of the altered alpha-helix to beta-sheet ratio with the mid-IR spectrum, as well as differentiation between the treatments using PCA and CLA. PCA and CLA results suggest that mid-IR spectral methods are more sensitive than traditional methods when used to identify differences between the heat treatments.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-27T13:55:23Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Danilo Marques Leal - 2014.pdf: 1178429 bytes, checksum: 4e858544ab13c00a2670e3bfc26568ae (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-01-28T11:52:43Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Danilo Marques Leal - 2014.pdf: 1178429 bytes, checksum: 4e858544ab13c00a2670e3bfc26568ae (MD5) / Made available in DSpace on 2015-01-28T11:52:44Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Dissertação - Danilo Marques Leal - 2014.pdf: 1178429 bytes, checksum: 4e858544ab13c00a2670e3bfc26568ae (MD5) Previous issue date: 2014-03-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / (Sem resumo em outra língua) / A presente pesquisa teve como objetivo avaliar a produtividade e composição bromatológica do cultivar Mulato II, sob regime de cortes e submetido a doses de nitrogênio, no município de Goiânia, Goiás. Utilizou-se um delineamento experimental inteiramente casualizado com arranjo fatorial 2 X 4 (2 alturas de corte X 4 doses de nitrogênio) com três repetições e parcelas subdivididas. Os tratamentos foram constituídos por quatro doses de N (sendo a fonte ureia) (0, 50, 100 e 150 kg/ha de N), com duas alturas de entrada (0,40 e 0,50 m). Todos os cortes foram realizados no período compreendido entre 05 de dezembro de 2012 e 24 de abril de 2013, e o parâmetro utilizado para a realização dos cortes foi a altura do dossel, conforme cada tratamento. Não ocorreu interação significativa (p>0,05) entre doses de N e alturas de corte para as variáveis PMV, PMS, ECAN e RAN. As produtividades de massa verde (PMV) e massa seca (PMS), a eficiência de conversão aparente de nitrogênio (ECAN) e a recuperação aparente de nitrogênio (RAN) não foram influenciadas (p<0,05) pelas doses de N avaliadas (0, 50, 100 e 150 kg.ha-1) nem em função das alturas de corte avaliadas (0,40 e 0,50 m). A média de produtividade encontrada foi de 59.450 kg.ha-1 (PMV) e 10.367 kg.ha-1 (PMS), produzindo em média 19,62 kg de MS para cada kg de N aplicado, com uma recuperação média de 56,00%. As doses de N aplicadas e as alturas de corte não influenciaram (p>0,05) os teores de MS da planta. A média do teor de matéria seca encontrada foi de 17,49%. Os teores de PB foram influenciados (p<0,05) pelas doses de N (0, 50, 100 e 150 kg.ha-1) e alturas de corte (0,40 e 0,50 m) bem como a interação desses fatores. Os teores de proteína bruta (PB) da planta inteira Brachiaria híbrida cv. Mulato II aumentaram (p<0,05) em função das doses de N, apresentando uma relação linear crescente. Quanto à altura de corte, ocorreu também efeito significativo (p<0,05), evidenciando que os teores de PB diminuíram com o aumento da altura. Os teores de FDN foram influenciados significativamente pelas doses de N (p<0,05) e pelas alturas de corte (p<0,05), apresentando significância para a interação (p<0,05) apenas nas doses acima de 100 kg.ha-1 de N. Para os teores de FDA não houve interação significativa (p>0,05) entre as variáveis analisadas. O conteúdo de FDA foi influenciado pelo fornecimento de N (p<0,05), apresentando regressão quadrática decrescente com o aumento das doses de N, em que a dose que proporciona o menor teor de FDA é de aproximadamente 226 kg.ha-1 de N. Ocorreram diferenças significativas (p<0,05) para CT, entre as doses de N e entre as alturas de corte, sendo observadas interações significativas (p<0,05) entre doses de N X alturas de corte, para doses acima de 50 kg.ha-1. Para as frações de carboidratos, foram observadas diferenças estatísticas (p<0,05) para as frações A + B1 e B2, não sendo encontradas diferenças para a fração C. Houve aumento linear para as frações A + B1 em resposta ao aumento das doses de N. Em relação a fração B2, o aumento das frações A+B1, resultaram em efeito linear decrescente desta. A fração C não foi influenciada pela adubação nitrogenada, nem para as alturas de corte, apresentando valores médios de 8,90%. Para as frações de compostos nitrogenados não foram observadas diferenças significativas (p>0,05) entre as frações A de proteínas, para nenhum dos tratamentos. A fração B1+B2 apresentou diferença (p<0,05) entre doses de N apenas para a altura de 0,50 m. Foi observada diferença significativa (p<0,05) entre doses de N e alturas de corte para as frações B3 e C.