Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis
Ano de defesa: | 2010 |
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Autor(a) principal: | |
Orientador(a): | |
Banca de defesa: | , |
Tipo de documento: | Dissertação |
Tipo de acesso: | Acesso aberto |
Idioma: | por |
Instituição de defesa: |
Universidade Federal de Santa Maria
|
Programa de Pós-Graduação: |
Programa de Pós-Graduação em Zootecnia
|
Departamento: |
Zootecnia
|
País: |
BR
|
Palavras-chave em Português: | |
Palavras-chave em Inglês: | |
Área do conhecimento CNPq: | |
Link de acesso: | http://repositorio.ufsm.br/handle/1/10753 |
Resumo: | In vitro assays were conducted to develop a method to estimate activity of bacterial enzymes in the rumen and the effect of changes in pH (5.5, 6.0, 6.5 and 7.0) and concentration of soluble carbohydrates (0, 100, 200 and 300 mg / dl) on adhesion to forage particles and ruminal bacterial fibrolytic activity. To evaluate the enzymatic extraction methods, samples of ryegrass (Lolium multiflorum) were incubated in a bovine rumen during 24-h and used to obtain enzymatic extracts through different extraction methods: sonication, sonication + lyzozyme, lyzozyme and without previous treatment (control). Then, the enzymatic extract was incubated with carboxymethylcelullose (CMC) and measured the reducing liberated sugars. The enzymatic extraction method did not affect significantly the enzymatic activity, but the standard deviation was lower when extraction only was through sonication. In another methodological assay, the effect of the reaction time between enzymatic extract and substrate on relative enzymatic activity (nmol of reducing sugars/ g DM of sample residue/ min) was tested. The relative activity decreased exponentially with time and stabilized after 240-min of incubation. To evaluate the effects of pH and concentration of sugars on bacterial attachment to feed particles and his degradability, samples of ryegrass (Lolium multiflorum) or cynodon (Cynodon dactilon) were incubated in vitro for 24-h in mediums with different pH (5,5; 6,0; 6,5 or 7,0) or different initial concentrations of glucose (0, 100, 200 and 300 mg/dl). The DM and phosphorus content (phosphorus was assumed be of bacterial origin) were determined in the fermentation residue. In ryegrass samples degradability and the residual phosphorus content were higher than in cynodon (P<0.01) and both factors were affected linearly and positively by pH (<0.01). The increase of soluble carbohydrates concentration exerted linear and negative effects on bacterial attachment (P<0.01), but it did not affected forage degradability. The effect of pH and sugar concentration was evaluated on fibrolityc enzyme activity in residues of the samples incubated in the rumen for 24-h and extracted through sonication. The enzymatic extract was incubated for 240-min with xylan or CMC in reaction mediums with different pH or sugar concentrations. In average, the activity of the enzymes extracted from the ryegrass residue was superior to the cynodon ones (P<0.01). On the other hand, the effect of both factors varied among assays and with the sample type, e.g. in different assays was observed that the pH effect was linear (P<0.05), quadratic (P<0.05) or even do not adjusted to none of the regressions. In the same way, the effect of presence of different concentrations of sugars in the medium on enzyme activity was quadratic mainly (P<0.01). In conclusion, the bacterial attachment to the forage particles was related directly to pH and inversely with concentration of soluble sugars in the incubation medium. However, these effects were less evident on enzymatic fibrolytic activity. |
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2010-06-072010-06-072010-02-26FARENZENA, Roberta. Adherence and bacterial enzyme activity in the rumen: effect of pH and concentration of soluble carbohydrates. 2010. 101 f. Dissertação (Mestrado em Zootecnia) - Universidade Federal de Santa Maria, Santa Maria, 2010.http://repositorio.ufsm.br/handle/1/10753In vitro assays were conducted to develop a method to estimate activity of bacterial enzymes in the rumen and the effect of changes in pH (5.5, 6.0, 6.5 and 7.0) and concentration of soluble carbohydrates (0, 100, 200 and 300 mg / dl) on adhesion to forage particles and ruminal bacterial fibrolytic activity. To evaluate the enzymatic extraction methods, samples of ryegrass (Lolium multiflorum) were incubated in a bovine rumen during 24-h and used to obtain enzymatic extracts through different extraction methods: sonication, sonication + lyzozyme, lyzozyme and without previous treatment (control). Then, the enzymatic extract was incubated with carboxymethylcelullose (CMC) and measured the reducing liberated sugars. The enzymatic extraction method did not affect significantly the enzymatic activity, but the standard deviation was lower when extraction only was through sonication. In another methodological assay, the effect of the reaction time between enzymatic extract and substrate on relative enzymatic activity (nmol of reducing sugars/ g DM of sample residue/ min) was tested. The relative activity decreased exponentially with time and stabilized after 240-min of incubation. To evaluate the effects of pH and concentration of sugars on bacterial attachment to feed particles and his degradability, samples of ryegrass (Lolium multiflorum) or cynodon (Cynodon dactilon) were incubated in vitro for 24-h in mediums with different pH (5,5; 6,0; 6,5 or 7,0) or different initial concentrations of glucose (0, 100, 200 and 300 mg/dl). The DM and phosphorus content (phosphorus was assumed be of bacterial origin) were determined in the fermentation residue. In ryegrass samples degradability and the residual phosphorus content were higher than in cynodon (P<0.01) and both factors were affected linearly and positively by pH (<0.01). The increase of soluble carbohydrates concentration exerted linear and negative effects on bacterial attachment (P<0.01), but it did not affected forage degradability. The effect of pH and sugar concentration was evaluated on fibrolityc enzyme activity in residues of the samples incubated in the rumen for 24-h and extracted through sonication. The enzymatic extract was incubated for 240-min with xylan or CMC in reaction mediums with different pH or sugar concentrations. In average, the activity of the enzymes extracted from the ryegrass residue was superior to the cynodon ones (P<0.01). On the other hand, the effect of both factors varied among assays and with the sample type, e.g. in different assays was observed that the pH effect was linear (P<0.05), quadratic (P<0.05) or even do not adjusted to none of the regressions. In the same way, the effect of presence of different concentrations of sugars in the medium on enzyme activity was quadratic mainly (P<0.01). In conclusion, the bacterial attachment to the forage particles was related directly to pH and inversely with concentration of soluble sugars in the incubation medium. However, these effects were less evident on enzymatic fibrolytic activity.Ensaios in vitro foram conduzidos com o objetivo de desenvolver um método para estimar atividade de enzimas bacterianas do rúmen e avaliar os efeitos da alteração no pH e na concentração de carboidratos solúveis sobre a aderência às partículas de forragem e sobre a atividade fibrolítica bacteriana ruminal. Para avaliação de métodos de extração enzimática, amostras de azevém (lolium multiflorum) foram incubadas no rúmen de um bovino durante 24 horas e utilizadas para obtenção de extratos enzimáticos por diferentes métodos de extração: sonicação, sonicação + lisozima, lisozima e sem tratamento prévio (controle). O extrato enzimático foi então incubado com carboximetilcelulose (CMC) e medido a liberação de açúcares redutores. O método de extração de enzimas não afetou significativamente a atividade enzimática, mas o desvio padrão foi menor quando a extração foi somente por sonicação. Em outro ensaio metodológico foi testado o efeito do tempo de reação entre extrato enzimático e substrato sobre a atividade relativa das enzimas (nmol de açúcares redutores/g de matéria seca de resíduo de amostra/minuto). A atividade relativa diminuiu exponencialmente com o tempo, estabilizando-se a partir de 240 minutos de incubação. Para avaliar o efeito do pH e da concentração de açúcares sobre a aderência bacteriana às partículas de alimento e sobre a sua degradabilidade, amostras de azevém (Lolium multiflorum) ou cynodon (Cynodon dactilon) foram incubadas in vitro durante 24h em meios com diferentes pHs (5,5; 6,0; 6,5 ou 7,0) ou diferentes concentrações iniciais de glicose (0, 100, 200 e 300 mg/dl). No resíduo da fermentação foi determinado o teor de matéria seca e analisado o teor de P, assumindo-se ser de origem bacteriana. A degradabilidade e o teor de P residual foram mais altos nas amostras de azevém que de cynodon (P<0,01) e ambos fatores foram linearmente e positivamente afetados pelo pH (P<0,01). O aumento da concentração de carboidratos solúveis exerceu efeito linear e negativo sobre a aderência das bactérias (P<0,01), mas não afetou a degradação da forragem. O efeito do pH e da concentração de açúcares sobre a atividade enzimática fibrolítica foi avaliada no resíduo destas amostras forrageiras incubadas ruminalmente durante 24h e extraídas por sonicação. O extrato enzimático foi incubado durante 240 minutos com xilana ou CMC em meios de reação a diferentes pHs ou concentração de açúcares. Em média, a atividade de enzimas extraídas de resíduo de azevém foi superior à atividade daquelas oriundas de cynodon (P<0,01). No entanto, o efeito de ambos fatores variou entre ensaios e com o tipo de amostra. Por exemplo, em diferentes ensaios foi observado que o efeito de pH foi linear (P<0,05), quadrático (P<0,05) ou mesmo não foi ajustado à nenhuma destas regressões. Da mesma forma, a presença de diferentes concentrações de açúcares no meio exerceu efeito, na maioria das vezes quadrático, sobre a atividade enzimática (P<0,01). Em conclusão, a aderência bacteriana às partículas de forragem foi diretamente relacionada com o pH e inversamente relacionada com a concentração de açúcares solúveis no meio de incubação. Estes efeitos, contudo, foram menos evidentes sobre a atividade enzimática fibrolítica.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em ZootecniaUFSMBRZootecniaAçúcaresAderênciaBactérias ruminaisDegradaçãoEnzimasFibraPHAdhesionDegradationEnzymesFiberPHRumen bacteriaSugarsCNPQ::CIENCIAS AGRARIAS::ZOOTECNIAAderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveisAdherence and bacterial enzyme activity in the rumen: effect of pH and concentration of soluble carbohydratesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisKozloski, Gilberto Vilmarhttp://lattes.cnpq.br/1966801022255836Perottoni, Julianohttp://lattes.cnpq.br/7042204446342433Rocha, João Batista Teixeira dahttp://lattes.cnpq.br/3935055744673018http://lattes.cnpq.br/3702518212441044Farenzena, Roberta500400000002400500500300500b4a3eff6-7fb0-444e-9501-ff96d450f4c9c94f98aa-37bc-4a2e-b30c-9b7b4c5f97d69bbc678c-7ad7-46a2-bc8c-0af75fdce1a9287bb0df-2738-40f7-8e03-0d36bf19a982info:eu-repo/semantics/openAccessreponame:Biblioteca Digital de Teses e Dissertações do UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALDIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdfDissertação de Mestradoapplication/pdf400186http://repositorio.ufsm.br/bitstream/1/10753/1/DIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf6b0c18a48d0f91c37cf33b53ff1b70d6MD51TEXTFARENZENA, ROBERTA.pdf.txtFARENZENA, ROBERTA.pdf.txtExtracted texttext/plain147735http://repositorio.ufsm.br/bitstream/1/10753/2/FARENZENA%2c%20ROBERTA.pdf.txt035eb1fc129a122c7062903d85c6842bMD52DIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.txtDIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.txtExtracted texttext/plain147735http://repositorio.ufsm.br/bitstream/1/10753/4/DIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.txt035eb1fc129a122c7062903d85c6842bMD54THUMBNAILFARENZENA, ROBERTA.pdf.jpgFARENZENA, ROBERTA.pdf.jpgIM Thumbnailimage/jpeg4840http://repositorio.ufsm.br/bitstream/1/10753/3/FARENZENA%2c%20ROBERTA.pdf.jpg1f3e6203df59fc9f67a5e69102291c02MD53DIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.jpgDIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.jpgIM Thumbnailimage/jpeg4840http://repositorio.ufsm.br/bitstream/1/10753/5/DIS_PPGZOOTECNIA_2010_FARENZENA_ROBERTA.pdf.jpg1f3e6203df59fc9f67a5e69102291c02MD551/107532022-04-11 16:32:27.886oai:repositorio.ufsm.br:1/10753Biblioteca Digital de Teses e Dissertaçõeshttps://repositorio.ufsm.br/ONGhttps://repositorio.ufsm.br/oai/requestatendimento.sib@ufsm.br||tedebc@gmail.comopendoar:2022-04-11T19:32:27Biblioteca Digital de Teses e Dissertações do UFSM - Universidade Federal de Santa Maria (UFSM)false |
dc.title.por.fl_str_mv |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
dc.title.alternative.eng.fl_str_mv |
Adherence and bacterial enzyme activity in the rumen: effect of pH and concentration of soluble carbohydrates |
title |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
spellingShingle |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis Farenzena, Roberta Açúcares Aderência Bactérias ruminais Degradação Enzimas Fibra PH Adhesion Degradation Enzymes Fiber PH Rumen bacteria Sugars CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA |
title_short |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
title_full |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
title_fullStr |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
title_full_unstemmed |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
title_sort |
Aderência e atividade fibrolítica bacteriana ruminal: efeito do pH e da concentração de carboidratos solúveis |
author |
Farenzena, Roberta |
author_facet |
Farenzena, Roberta |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Kozloski, Gilberto Vilmar |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1966801022255836 |
dc.contributor.referee1.fl_str_mv |
Perottoni, Juliano |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/7042204446342433 |
dc.contributor.referee2.fl_str_mv |
Rocha, João Batista Teixeira da |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/3935055744673018 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/3702518212441044 |
dc.contributor.author.fl_str_mv |
Farenzena, Roberta |
contributor_str_mv |
Kozloski, Gilberto Vilmar Perottoni, Juliano Rocha, João Batista Teixeira da |
dc.subject.por.fl_str_mv |
Açúcares Aderência Bactérias ruminais Degradação Enzimas Fibra PH |
topic |
Açúcares Aderência Bactérias ruminais Degradação Enzimas Fibra PH Adhesion Degradation Enzymes Fiber PH Rumen bacteria Sugars CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA |
dc.subject.eng.fl_str_mv |
Adhesion Degradation Enzymes Fiber PH Rumen bacteria Sugars |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA |
description |
In vitro assays were conducted to develop a method to estimate activity of bacterial enzymes in the rumen and the effect of changes in pH (5.5, 6.0, 6.5 and 7.0) and concentration of soluble carbohydrates (0, 100, 200 and 300 mg / dl) on adhesion to forage particles and ruminal bacterial fibrolytic activity. To evaluate the enzymatic extraction methods, samples of ryegrass (Lolium multiflorum) were incubated in a bovine rumen during 24-h and used to obtain enzymatic extracts through different extraction methods: sonication, sonication + lyzozyme, lyzozyme and without previous treatment (control). Then, the enzymatic extract was incubated with carboxymethylcelullose (CMC) and measured the reducing liberated sugars. The enzymatic extraction method did not affect significantly the enzymatic activity, but the standard deviation was lower when extraction only was through sonication. In another methodological assay, the effect of the reaction time between enzymatic extract and substrate on relative enzymatic activity (nmol of reducing sugars/ g DM of sample residue/ min) was tested. The relative activity decreased exponentially with time and stabilized after 240-min of incubation. To evaluate the effects of pH and concentration of sugars on bacterial attachment to feed particles and his degradability, samples of ryegrass (Lolium multiflorum) or cynodon (Cynodon dactilon) were incubated in vitro for 24-h in mediums with different pH (5,5; 6,0; 6,5 or 7,0) or different initial concentrations of glucose (0, 100, 200 and 300 mg/dl). The DM and phosphorus content (phosphorus was assumed be of bacterial origin) were determined in the fermentation residue. In ryegrass samples degradability and the residual phosphorus content were higher than in cynodon (P<0.01) and both factors were affected linearly and positively by pH (<0.01). The increase of soluble carbohydrates concentration exerted linear and negative effects on bacterial attachment (P<0.01), but it did not affected forage degradability. The effect of pH and sugar concentration was evaluated on fibrolityc enzyme activity in residues of the samples incubated in the rumen for 24-h and extracted through sonication. The enzymatic extract was incubated for 240-min with xylan or CMC in reaction mediums with different pH or sugar concentrations. In average, the activity of the enzymes extracted from the ryegrass residue was superior to the cynodon ones (P<0.01). On the other hand, the effect of both factors varied among assays and with the sample type, e.g. in different assays was observed that the pH effect was linear (P<0.05), quadratic (P<0.05) or even do not adjusted to none of the regressions. In the same way, the effect of presence of different concentrations of sugars in the medium on enzyme activity was quadratic mainly (P<0.01). In conclusion, the bacterial attachment to the forage particles was related directly to pH and inversely with concentration of soluble sugars in the incubation medium. However, these effects were less evident on enzymatic fibrolytic activity. |
publishDate |
2010 |
dc.date.accessioned.fl_str_mv |
2010-06-07 |
dc.date.available.fl_str_mv |
2010-06-07 |
dc.date.issued.fl_str_mv |
2010-02-26 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FARENZENA, Roberta. Adherence and bacterial enzyme activity in the rumen: effect of pH and concentration of soluble carbohydrates. 2010. 101 f. Dissertação (Mestrado em Zootecnia) - Universidade Federal de Santa Maria, Santa Maria, 2010. |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufsm.br/handle/1/10753 |
identifier_str_mv |
FARENZENA, Roberta. Adherence and bacterial enzyme activity in the rumen: effect of pH and concentration of soluble carbohydrates. 2010. 101 f. Dissertação (Mestrado em Zootecnia) - Universidade Federal de Santa Maria, Santa Maria, 2010. |
url |
http://repositorio.ufsm.br/handle/1/10753 |
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por |
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Universidade Federal de Santa Maria |
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