Thesis (PhD(Agric))--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Exogenous fibrolytic enzymes (EFE) as additives in ruminant feeds are being researched worldwide. Promising effects on dry matter intake (DMI), digestibility and production in especially dairy cows, but also feedlot steers and even sheep have been observed. However, lack of or negative effects are also reported and the need arises for clarity on the mode-of-action of EFE. Forages are characterised as being highly heterogenic and contain varying concentrations of fibre. The fibre, in turn, varies greatly in digestibility, due to the chemical as well as anatomical build-up of this complex carbohydrate. Fibre, however, presents a major source of potential energy for ruminant animals and EFE is a viable option to increase the digestibility of forages. Therefore, a study with the aim of establishing whether EFE can affect the digestibility of forages, how it affects the digestibility and the clarification of the mode-of-action was drafted. From the literature, the first objective was readily attained and clear indications exist that EFE can indeed improve animal performance when their diets are treated with such enzymes. From the current study, it was shown that EFE can alter the rate and extent of gas production of certain forages (lucerne, kikuyu and weeping love grass) and also improve the in vitro digestibility thereof (P < 0.05). This is in agreement with other research findings and it was concluded that these effects were likely exerted during the early stages of digestion. A complete feed for sheep, when treated with the EFE, showed positive effects on the in sacco digestibility, as well as on the digestion kinetics of the feed (P < 0.05). The in vitro digestibility of the complete feed was also improved due to EFE treatment (P < 0.05). The observations on in vitro digestibility were less marked when a purified xylanase, obtained from the partial purification of the EFE cocktail, was used as the sole fibrolytic enzyme treatment. It is apparent, therefore, that enzyme specificity plays a major role in obtaining positive effects on digestibility of forages and feeds. In agreement with the literature, it is proposed that the approach to improve the digestibility of forages should be to use EFE cocktails containing various enzymes, matching the complexity of the substrate. The major aim of the study was, however, an in depth investigation of the mode-of-action of EFE. This aim was approached by observing changes in plant tissue material at the histological level upon treatment with EFE and incubation in buffered rumen fluid. Results showed that EFE had subtle, yet significant effects on cell wall material for the various tissues studied (P < 0.05). The major effect observed here was that EFE had a thinning effect on the cell wall thickness (P < 0.05). It was deduced that as EFE affected the cell wall of the plant material, earlier access by microorganisms could be achieved. Also, nutrients caught in the cell wall matrix could then be released for digestion. Therefore, observations that EFE increases the rate of digestion, as well as the extent of digestion of not only fibre, but also protein, were explained by the enzyme’s action on cell wall material. It was concluded that there is definite merit in the use of EFE to improve the digestibility of ruminant feeds and that this is partly related to effects on the cell walls of the forages. The effects can be expected to occur during the early stages of digestion, thereby potentially increasing the passage rate of digesta from the rumen. Additionally, the effect of the EFE is not limited to fibre and increased digestibility of all nutrients can be expected, thereby increasing the overall digestibility of the feed. Future research should further elucidate the mode-of-action of EFE using advanced technologies routinely employed in the plant sciences. Additionally, the main potential advantage of EFE treatment lies in improving the digestibility of poor quality roughages. Unfortunately, this is an area where limited positive effects are observed and in depth investigations should be undertaken to classify the specificity and optimum conditions of EFE to better match the complexity of the substrate being treated. / AFRIKAANSE OPSOMMING: Eksogene fibrolitiese ensieme (EFE) word tans wêreldwyd ondersoek vir die gebruik daarvan as voerbymiddels vir herkouers. Belowende effekte op DMI, verteerbaarheid en produksie van vernaam melkbeeste, maar ook voerkraalbeeste en selfs skape is al gerapporteer. Swak en selfs negatiewe effekte word egter ook waargeneem en daarom is ʼn deeglike ondersoek na die metode van werking van EFE van belang. Ruvoere word gekenmerk deurdat dit heterogeen van aard is en bevat variërende vlakke van vesel. Vesel maak op sy beurt ʼn wesenlike deel uit van die ruvoer, maar varieer baie in verteerbaarheid weens die chemiese sowel as anatomiese samestelling van hierdie komplekse koolhidraat. Ruvoer verteenwoordig egter ʼn goeie bron van potensiële energie vir herkouers en EFE word voorgestel as ʼn haalbare behandeling om die verteerbaarheid daarvan te verhoog. Dus is ʼn studie beplan met die doelwit om die effekte van EFE te definieer, hoe dit verteerbaarheid beïnvloed en die metode van werking daarvan te ondersoek. Vanuit die literatuur is dit duidelik dat daar wel baie positiewe effekte is waar ruvoere met EFE behandel is en dat diereproduksie wel bevoordeel word daardeur. Vanuit die studie is dit getoon dat die tempo en hoeveelheid gasproduksie van sekere ruvoere (lusern, kikuyu en oulandsgras) verbeter word deur EFE behandeling (P < 0.05). Hierdie bevinding was ondersteun deur verbeterde in vitro verteerbaarheid van die ruvoere (P < 0.05). Dit is in ooreenstemming met literatuur en die gevolgtrekking is gemaak dat hierdie effekte tydens die vroeëre stadia van vertering verwag kan word. ʼn Volledige skaapvoer wat met EFE behandel is, het positiewe effekte op in sacco verteerbaarheid en verterings kinetika data gehad (P < 0.05). Weereens is die in vitro verteerbaarheid van die voer verbeter (P < 0.05). Waarnemings op in vitro verteerbaarheid was veel minder opvallend wanneer ʼn gesuiwerde xylanase as enigste fibrolitiese ensiem behandeling gebruik is. Dit is dus duidelik dat ensiem spesifisiteit ʼn belangrike rol speel in die verkryging van positiewe resultate in die verteerbaarheid van ruvoere en veevoere. In ooreenstemming met literatuur word dit voorgestel dat ensiemmengsels wat verskeie ensieme bevat as EFE gebruik behoort te word ten einde aan die kompleksiteit van die substraat te voldoen. Die hoof doelwit van die studie was egter ʼn indiepte ondersoek na die metode van werking van EFE. Hierdie doelwit is benader deur die effekte van EFE op selwand strukture van plantmateriaal op ʼn histologiese vlak te ondersoek. Die ruvoere was vooraf met EFE behandel en in vitro geïnkubeer in rumen vloeistof. Die resultate het getoon dat EFE ʼn matige, dog betekenisvolle effek op die selwand materiaal van die onderskeie weefsels gehad het (P < 0.05). Die belangrikste waargeneemde effek was dat EFE ʼn verdunningseffek op die selwande gehad het. Dit is afgelei dat as EFE die selwand kan beïnvloed, mikro-organismes vroeër toegang tot die inhoud kan kry. Verder, nutriënte vasgevang in die selwand matriks raak ook beskikbaar vir vertering. Hierdie afleiding en die effek van EFE op selwande verklaar waarnemings dat EFE die tempo van vertering sowel as die vlak van vertering van nie net vesel, maar ook proteïen kan bevoordeel. Die gevolgtrekking is gemaak dat daar definitiewe meriete is in die gebruik van EFE om die verteerbaarheid van herkouervoere te verbeter en dat dit verband hou met die ensiem se werking op selwande van die ruvoere. Die effekte kan verwag word tydens die vroeë stadia van vertering om dus deurvloeitempo van digesta te verbeter. Die effek van die EFE is verder nie beperk tot vesel nie, maar positiewe effekte op ander nutriënte kan verwag word en vervolgens ʼn algehele verhoging in die verteerbaarheid van die voer. Navorsing behoort in die toekoms verder die metode van werking van EFE te ondersoek deur gebruik te maak van gevorderde tegnologie wat alledaags gebruik word in die Plantwetenskappe. Die belowendste aanwending van EFE lê in die verbetering in vertering van swak kwaliteit ruvoere. Dit is ongelukkig juis hier waar min positiewe resultate gerapporteer word en indiepte navorsing moet onderneem word om ensiem spesifisiteit en optimum kondisies te definieer sodat EFE beter opgewasse is teen die kompleksiteit van die substraat.

A preocupação com a segurança alimentar e o aquecimento global pressiona a cadeia de produção de ruminantes a buscar alternativas para incremento na produtividade e redução nas emissões dos gases de efeito estufa, uma vez que essa atividade é responsável por grande parte das emissões de metano (CH4) entérico. O desenvolvimento de produtos enzimáticos para uso na nutrição de ruminantes torna-se de grande valia para contribuir com a demanda alimentar e reduzir os impactos ambientais. O objetivo do presente estudo foi avaliar o uso de um extrato de enzimas fibrolíticas (EEF), composto por endoglucanases, exoglucanases e xilanases, produzido pelo fungo Trichoderma reesei utilizando como substrato resíduos da indústria canavieira, sobre a digestibilidade, fermentação ruminal e desempenho de ovinos em crescimento, conduzidos através de dois experimentos. No experimento 1, foi avaliado a adição de doses crescentes (0, 10, 100, 1000 e 10000 µL g-1 de substrato) do EEF sobre a produção total de gases e CH4, degradabilidade e fermentação ruminal de três forrageiras tropicais (Cynodon spp., Panicum maximum e Cenchrus ciliares L.) em um ensaio in vitro, com o objetivo de verificar a possibilidade de uso do EEF na dieta de ruminantes sem prejudicar os parâmetros fermentativos. Verificou-se que os resultados foram dependentes da dose e do substrato utilizados. Contudo, foi verificado aumento na produção total de gases e CH4 e na degradabilidade de todas as forrageiras utilizadas com a dose mais alta do EEF. Conclui-se que o EEF pode ser utilizado na dieta de ruminantes como alternativa para incremento na degradabilidade de forrageiras tropicais. O experimento 2 foi dividido em duas fases conduzidas com o objetivo de avaliar o efeito do EEF sobre a digestibilidade aparente dos nutrientes, fermentação ruminal, desempenho animal e produção de CH4 entérico de 19 ovinos em crescimento divididos em dois grupos experimentais, controle (CTL) e enzima (ENZ), recebendo dieta composta de 70% de feno de Tifton-85 e 30% de mistura concentrada. O EEF foi aplicado diretamente no feno 24 horas antes do fornecimento da refeição. Na Fase I, avaliou-se o efeito de doses crescentes (0, 20, 40 e 80 µL) do EEF sobre a hidrólise da fibra e posteriormente a dose com melhor resposta foi utilizada nos estudos de digestibilidade aparente dos nutrientes e fermentação ruminal. A dose de 40 µL foi selecionada para ser utilizada no ensaio in vivo, pois apresentou melhores respostas sobre a redução nas frações fibrosas. Os animais suplementados com o EEF apresentaram maior digestibilidade da fibra em detergente ácido e maior produção de valerato. Na Fase II, foi avaliado o efeito do EEF sobre o desempenho, saúde e produção de CH4 entérico de ovinos em crescimento. Os procedimentos utilizados foram os mesmos descritos na Fase I. A suplementação do EEF proporcionou maior percentual médio de ganho de peso e menor produção de CH4 entérico por kg de matéria seca ingerida. As análises sanguíneas demonstraram que o EEF não prejudicou a saúde dos animais. Conclui-se que o EEF pode ser utilizado na dieta animal sem trazer riscos à saúde, além de ser uma estratégia para aumento na produtividade animal e contribuir com a mitigação de CH4 entérico de ovinos em crescimento / Concern about food security and global warming is pressing the ruminant production chain to seek alternatives to increase productivity and reduce greenhouse gas emissions, as this activity accounts for most of the enteric methane (CH4) emissions. The development of enzyme products for use in ruminant nutrition is of great value in order to contribute to food demand and reduce environmental impacts. The aim of the present study was to evaluate the use of a fibrolytic enzyme extract (FEE), produced by the fungus Trichoderma reesei using as substrate residues from the sugar cane industry, on the digestibility, ruminal fermentation and performance of growing sheep, conducted through two experiments. In the experiment 1, the addition of increasing doses (0, 10, 100, 1000 and 10000 ?L g-1) of the FEE on the total gas and CH4 production, degradability and ruminal fermentation of three tropical forages (Cynodon spp., Panicum maximum and Cenchrus ciliares L.) in an in vitro assay, with the objective of verifying the possibility of use of the FEE in the diet of ruminants without harming the fermentative parameters and, consequently, animal health. The results were found to be dose and substrate dependent. However, there was an increase in total gas and CH4 production and in the degradability of all forages used with the highest dose of FEE. It is concluded that the EPS can be used in the diet of ruminants as an alternative to increase the degradability of tropical forages. The experiment 2 was divided in two phases, with the objective of evaluating the effect of the FEE on the apparent digestibility of nutrients, ruminal fermentation, animal performance and enteric CH4 production of 19 growing sheep divided in two experimental groups, control (CTL) and enzyme (ENZ), receiving a diet composed of 70% of Tifton-85 hay and 30% of concentrated mixture. The FEE was applied directly to the hay 24 hours before the meal. In Phase I, the effect of increasing doses (0, 20, 40 and 80 ?L) of the FEE on fiber hydrolysis was evaluated and the dose with the best response was then used in the studies of apparent digestibility of nutrients and ruminal fermentation. The 40 ?L dose was selected to be used in the in vivo assay because it presented better responses on the reduction in fibrous fractions. The animals supplemented with FEE showed higher acid digestible fiber digestibility and higher valerate production. The other parameters were not influenced by the addition of FEE. In Phase II, the effect of FEE was evaluated on the performance, health and production of Enteric CH4 of growing sheep. The procedures used were the same as described in Phase I. The supplementation of the FEE provided a higher percentage of weight gain and lower enteric CH4 production per kg of ingested dry matter. Blood tests showed that FEE did not affect animal health. It is concluded that FEE can be used in the animal diet without presenting health risks, besides being a strategy to increase animal productivity and contribute to the mitigation of enteric CH4 of growing sheep

A preocupação com a segurança alimentar e o aquecimento global pressiona a cadeia de produção de ruminantes a buscar alternativas para incremento na produtividade e redução nas emissões dos gases de efeito estufa, uma vez que essa atividade é responsável por grande parte das emissões de metano (CH4) entérico. O desenvolvimento de produtos enzimáticos para uso na nutrição de ruminantes torna-se de grande valia para contribuir com a demanda alimentar e reduzir os impactos ambientais. O objetivo do presente estudo foi avaliar o uso de um extrato de enzimas fibrolíticas (EEF), composto por endoglucanases, exoglucanases e xilanases, produzido pelo fungo Trichoderma reesei utilizando como substrato resíduos da indústria canavieira, sobre a digestibilidade, fermentação ruminal e desempenho de ovinos em crescimento, conduzidos através de dois experimentos. No experimento 1, foi avaliado a adição de doses crescentes (0, 10, 100, 1000 e 10000 µL g-1 de substrato) do EEF sobre a produção total de gases e CH4, degradabilidade e fermentação ruminal de três forrageiras tropicais (Cynodon spp., Panicum maximum e Cenchrus ciliares L.) em um ensaio in vitro, com o objetivo de verificar a possibilidade de uso do EEF na dieta de ruminantes sem prejudicar os parâmetros fermentativos. Verificou-se que os resultados foram dependentes da dose e do substrato utilizados. Contudo, foi verificado aumento na produção total de gases e CH4 e na degradabilidade de todas as forrageiras utilizadas com a dose mais alta do EEF. Conclui-se que o EEF pode ser utilizado na dieta de ruminantes como alternativa para incremento na degradabilidade de forrageiras tropicais. O experimento 2 foi dividido em duas fases conduzidas com o objetivo de avaliar o efeito do EEF sobre a digestibilidade aparente dos nutrientes, fermentação ruminal, desempenho animal e produção de CH4 entérico de 19 ovinos em crescimento divididos em dois grupos experimentais, controle (CTL) e enzima (ENZ), recebendo dieta composta de 70% de feno de Tifton-85 e 30% de mistura concentrada. O EEF foi aplicado diretamente no feno 24 horas antes do fornecimento da refeição. Na Fase I, avaliou-se o efeito de doses crescentes (0, 20, 40 e 80 µL) do EEF sobre a hidrólise da fibra e posteriormente a dose com melhor resposta foi utilizada nos estudos de digestibilidade aparente dos nutrientes e fermentação ruminal. A dose de 40 µL foi selecionada para ser utilizada no ensaio in vivo, pois apresentou melhores respostas sobre a redução nas frações fibrosas. Os animais suplementados com o EEF apresentaram maior digestibilidade da fibra em detergente ácido e maior produção de valerato. Na Fase II, foi avaliado o efeito do EEF sobre o desempenho, saúde e produção de CH4 entérico de ovinos em crescimento. Os procedimentos utilizados foram os mesmos descritos na Fase I. A suplementação do EEF proporcionou maior percentual médio de ganho de peso e menor produção de CH4 entérico por kg de matéria seca ingerida. As análises sanguíneas demonstraram que o EEF não prejudicou a saúde dos animais. Conclui-se que o EEF pode ser utilizado na dieta animal sem trazer riscos à saúde, além de ser uma estratégia para aumento na produtividade animal e contribuir com a mitigação de CH4 entérico de ovinos em crescimento / Concern about food security and global warming is pressing the ruminant production chain to seek alternatives to increase productivity and reduce greenhouse gas emissions, as this activity accounts for most of the enteric methane (CH4) emissions. The development of enzyme products for use in ruminant nutrition is of great value in order to contribute to food demand and reduce environmental impacts. The aim of the present study was to evaluate the use of a fibrolytic enzyme extract (FEE), produced by the fungus Trichoderma reesei using as substrate residues from the sugar cane industry, on the digestibility, ruminal fermentation and performance of growing sheep, conducted through two experiments. In the experiment 1, the addition of increasing doses (0, 10, 100, 1000 and 10000 ?L g-1) of the FEE on the total gas and CH4 production, degradability and ruminal fermentation of three tropical forages (Cynodon spp., Panicum maximum and Cenchrus ciliares L.) in an in vitro assay, with the objective of verifying the possibility of use of the FEE in the diet of ruminants without harming the fermentative parameters and, consequently, animal health. The results were found to be dose and substrate dependent. However, there was an increase in total gas and CH4 production and in the degradability of all forages used with the highest dose of FEE. It is concluded that the EPS can be used in the diet of ruminants as an alternative to increase the degradability of tropical forages. The experiment 2 was divided in two phases, with the objective of evaluating the effect of the FEE on the apparent digestibility of nutrients, ruminal fermentation, animal performance and enteric CH4 production of 19 growing sheep divided in two experimental groups, control (CTL) and enzyme (ENZ), receiving a diet composed of 70% of Tifton-85 hay and 30% of concentrated mixture. The FEE was applied directly to the hay 24 hours before the meal. In Phase I, the effect of increasing doses (0, 20, 40 and 80 ?L) of the FEE on fiber hydrolysis was evaluated and the dose with the best response was then used in the studies of apparent digestibility of nutrients and ruminal fermentation. The 40 ?L dose was selected to be used in the in vivo assay because it presented better responses on the reduction in fibrous fractions. The animals supplemented with FEE showed higher acid digestible fiber digestibility and higher valerate production. The other parameters were not influenced by the addition of FEE. In Phase II, the effect of FEE was evaluated on the performance, health and production of Enteric CH4 of growing sheep. The procedures used were the same as described in Phase I. The supplementation of the FEE provided a higher percentage of weight gain and lower enteric CH4 production per kg of ingested dry matter. Blood tests showed that FEE did not affect animal health. It is concluded that FEE can be used in the animal diet without presenting health risks, besides being a strategy to increase animal productivity and contribute to the mitigation of enteric CH4 of growing sheep

Orientador: Pedro de Oliva Neto / Resumo: Desejando ao final do trabalho obter um bioprocesso integrado usando palha de cana-de-açúcar (PC), este trabalho teve início com a utilização desse substrato para produção de enzimas fribrolíticas, xilanases e celulases, em culturas axênicas, incluindo espécies de Trichoderma e Aspergillus. A triagem para o melhor produtor foi realizada em “shaker” em fermentação submersa. A cultura do fungo T. reesei QM9414 alcançou a melhor produção de enzimas, e em tanque agitado, utilizando um biorreator de 3 L, mostrou o mesmo perfil de produção (~90 U/mL, 0.6 FPU/mL para xilanase e celulases, respectivamente). Em relação a este resultado, a produção de enzimas para as misturas binárias e ternárias destes fungos foi menor, sendo que a melhor combinação, T. reesei QM 9414+A. fumigatus M51, alcançou 60 U/mL e 0.08 FPU/mL respectivamente. Com intuito de otimizar a produção de enzimas utilizando um mix de substratos: palha de cana, como principal componente, e o farelo de trigo e a polpa cítrica, como supostos indutores de atividade enzimática, foi realizado um delineamento de misturas do tipo D-optimal. O resultado da otimização da mistura dos substratos mostrou que o trigo e a polpa cítrica não tiveram um efeito indutivo na produção das enzimas tendo a palha de cana como principal substrato. A enzima xilanase foi caracterizada em seu pH e temperatura ótimos (pH 5, e 50 ºC respectivamente), bem como a estabilidade da enzima nestes parâmetros. Alguns íons e EDTA foram aplicados para determin... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In order to obtain an integrated bioprocess using Sugarcane Straw (SS), this work began with the use of this substrate for the fibrolytic enzymes production, xylanases and cellulases, in axenic fungal cultures, including Trichoderma and Aspergillus species. The screening for the best producer was performed in shaker under submerged fermentation. The T. reesei QM9414 culture achieved the best enzyme production, and in a stirred tank using a 3 L bioreactor showed the same production profile (~90 U/mL and 0.6 FPU/mL for xylanase and cellulase, respectively). Regarding this result, the enzyme production by binary and ternary mixtures of these fungi was lower, as example the best combination T. reesei QM 9414+A. fumigatus M51, reached 60 U/mL and 0.08 FPU/mL, respectively. Aiming optimize the enzyme production by a mix of substrates using SS as the main substrate, and wheat bran and citrus pulp as supposed enzyme inductors, a D-optimal mixture design was performed. The mixture substrates optimization showed that wheat bran and citrus pulp did not have an inductive effect on the enzymes production. The enzyme xylanase was characterized by its optimal pH and temperature (pH 5 and 50 ºC, respectively, as well as the stability of the enzyme in these parameters. Some ions and EDTA were applied to determine the xylanase stability under these conditions, and the ion Mn2+ was the best inductor, 49% (10 mM). The extract containing xylanases, produced under previous optimized conditions was... (Complete abstract click electronic access below) / Doutor

Thesis (MScAgric (Animal Sciences))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: A growth and digestibility trial was conducted comparing the effect of an experimental enzyme at three different inclusions. A negative control containing no enzyme additions and a positive control containing a proven commercial enzyme were compared in a maize-soybean diet noting the performance of broilers and the digestibility of the grower feed. The commercial enzyme was a granular product with a xylanase activity of 38114.29 nkat/g and the second enzyme (ABO374) was a liquid experimental product with a xylanase activity of 1426.86 nkat/ml. Five diets were used i.e. control basal diet without enzyme supplementation (negative control), basal diet supplemented with the commercial enzyme (positive control) and three basal diets supplemented with the test enzyme at various inclusion levels (ABO 50, ABO 100 and ABO 200). The positive control was supplemented with 200 g/ton of the commercial enzyme; ABO 50 was supplemented with ABO374 at an inclusion level of 2671 ml/ton, ABO 100 with 5342 ml/ton and ABO 200 with 10684 ml/ton. Supplementation with the test enzyme (ABO 50) significantly improved BW at 23 days of age by 4.6 % (1107.4 g vs 960.96 g) and at 37 days of age by 3.2 % (2311.75 g vs 2237.81 g) over the negative control. Body weight gain for the total period of the trial was significantly improved by 3.24 % (64.32 g/bird/day vs 62.24 g/bird/day) the test enzyme supplementation (ABO 50) when compared to the negative control. During the starter phase, test enzyme supplementation (ABO 50) led to an improvement of 4.58 % (1.25 vs 1.31) in FCR in comparison with the negative control. The FCR for the total trial obtained by the test enzyme supplementation was significantly lower than the FCR obtained by the positive control. The highest EPER obtained for this trail was by the test enzyme supplemented diets and this was significantly higher than the EPER obtained by the positive control. It is clear from this growth trial that the test enzyme (ABO374) at an inclusion level of 2671 ml/ton outperformed the commercial enzyme and that it has the potential to improve the production performance of broilers on a maizeSBM based diet. The total tract digestibility method and total collection method was used to conduct the digestibility trial. The total tract digestibility method measures the difference between the amounts of each nutrient consumed from the amounts of each nutrient excreted in faeces. Only apparent digestibilities are reported for the digestibility trial. Apparent digestibility does not take the endogenous protein fraction in the faeces into account. The endogenous protein fraction is derived from digestive enzymes and proteins from the intestinal walls that are secreted into the digestive tract. The grower negative control, positive control, ABO 50, ABO 100 and ABO 200 diets used in the production trial were also used in the digestibility trial. Supplementation with the test enzyme showed no significant improvements on the apparent digestibility of dry matter, organic material, ash, crude protein, metabolisable energy or crude fibre. No significant improvements in the apparent digestibility of the amino acids (threonine, arginine, valine, lysine, methionine, cysteine and isoleucine) were noticed either and thus the digestibility of the grower feed were not influenced by the addition of enzymes due to the supplementation of the test enzyme ABO374. Pelletisation of the grower diets could have lead to the inactivation of the enzyme due to the high temperature at which pelletisation takes place. Another possible reason why enzyme supplementation did not increase nutrient digestibility, may be that the breakdown of non-starch polysaccharides by the enzymes led to an increase in the concentration of oligosaccharides in the small intestine of the birds, thus leading to the decrease in nutrient absorption Key words: body weight, body weight gain, feed conversion ratio, European production efficiency ratio, maize, soybean meal, apparent digestibility, dry matter, organic material, ash, crude protein, metabolisable energy, crude fibre, xylanase, pellitisation. / AFRIKAANSE OPSOMMING: ‘n Groei en vertering studie was uitgevoer om die effek van ‘n eksperimentele ensiem wat teen drie verskillende insluitingsvlakke by ‘n basale dieët bygevoeg is te vergelyk met ‘n negatiewe kontrole wat geen ensiem bevat het nie en met ‘n positiewe kontrole wat ‘n kommersiële ensiem bevat in ‘n mielesojaboon oliekoek dieët op die produksie vermoë van braaikuikens en die verteerbaarheid van die groei voer. Die kommersiële ensiem was ‘n granulêre produk met ‘n xylanase aktiwiteit van 38114.29 nkat/g en die eksperimentele ensiem (ABO374) was ‘n vloeistof produk met ‘n xylanase aktiwiteit van 1426.86 nkat/ml. Vyf diëte was gebruik nl. ‘n basale dieët met geen ensiem byvoeging (negatiewe kontrole), basale dieët met die byvoeging van die kommersiële ensiem (positiewe kontrole) en drie basale diëte wat met die byvoeging van die eksperimentele ensiem teen drie verskillende insluitings vlakke (ABO 50, ABO 100 and ABO 200). Die kommersiële ensiem was by die positiewe kontrole bygevoeg met ‘n insluitings vlak van 200 g/ton, ABO374 was bygevoeg by ABO 50 met ‘n insluitings vlak van 2671 ml/ton, ABO 100 met 5342 ml/ton en ABO 200 met 10684 ml/ton. Die byvoeging van die eksperimentele ensiem (ABO 50) het gelei tot die betekenisvolle verbetering van die liggaamsmassa van die voëls by die ouderdom van 23 dae met 4.6 % (1107.4 g teenoor 960.96 g) en by die ouderdom van 37 dae met 3.2 % (2311.75 g teenoor 2237.81 g) teenoor die negatiewe kontrole. Liggaams massa toename vir die hele periode van die studie was betekenisvol verhoog met 3.24 % (64.32 g/kuiken/dag teenoor 62.24 g/kuilen/dag) met die byvoeging van die eksperimentele ensiem (ABO374) teenoor die negatiewe kontrole. Voeromset verhouding was betekenisvol verbeter met 4.58 % (1.25 teenoor 1.31) toe die kommersiële ensiem bygevoeg was teenoor die negatiewe kontrole. Die hoogste europese produksie effektiwiteits verhouding wat verkry is vir die hele studie periode is deur die byvoeging van die eksperimentele ensiem (ABO374). Hierdie groei studie dui dus duidelik aan dat die gebruik van die eksperimentele ensiem (ABO374) baie beter resultate as die kommersiële ensiem opgelewer het teen ‘n insluitings vlak van 2671 ml/ton, dus het ABO374 die potensiaal om die produksie potensiaal van braaikuikens op ‘n mielie-sojaboonoliekoek dieët te verbeter. Die totale spysverteringskanaal verteerbaarheid metode was gebruik om die verteerbaarheid studie uit te voer. Die totale spysverteringskanaal verteerbaarheid metode meet die verskil tussen die nutriënt inhoud van die voer en die nutriënt inhoud van die mis. Slegs die skynbare verteerbaarheid van nutriënte word vir hierdie verteerbaarheidstudie gerapporteer. Skynbare verteerbaarheid sluit nie die endogene proteïenfraksie wat afkomstig is van verteringsensieme of die proteïene afkomstig vanaf die spysverteringskanaal se intestinale wande af in nie. Die negatiewe kontrole, positiewe kontrole, ABO 50, ABO 100 en ABO 200 groei diëte gebruik in die produksie studie is gebruik vir die verteringsstudie. Die byvoeging van die eksperimentele ensiem het geen betekenisvolle resultate opgelewer ten opsigte van droë materiaal, organiese material, as, ru-proteïen, ru-vesel of metaboliseerbare energie nie. Daar was ook geen betekenisvolle resultate opgelewer wanneer die eksperimentele ensiem bygevoeg was nie ten opsigte van die verteerbaarheid vir aminosure (treonien, arginien, valien, metionien, sisteïen en isoleosien) nie en dus is die verteerbaarheid van die groeivoer glad nie beïnvloed deur die byvoeging van die eksperimentele ensiem nie. Die verpilling van die groei voer mag dalk gelei het tot die inaktivering van die eksperimentele ensiem deur dat dit blootgestel was aan hoë temperature. ‘n Ander moontlike rede vir die mislukking van die ensiem kon gewees het dat die afbreking van die nie-stysel polisakkariedes deur die ensiem kon gelei het tot die verhoging van die oligosakkariede konsentrasie in die laer spysverterings kanaal en dus kon dit lei tot ‘n verhoogde deurvloeitempo, gevolg deur ‘n afname in die absorpsie van nutriënte. Sleutel woorde: Liggaamsmassa, liggaamsmassa toename, voeromsetverhouding Europese produksie effektiwiteits verhouding, mielie, sojaboonoliekoek, skynbare verteerbaarheid, droëmaterial, organiesematerial, as, ru-proteïen, ru-vesel, metaboliseerbare energie, verpilling.

Made available in DSpace on 2015-03-26T13:55:23Z (GMT). No. of bitstreams: 1 texto completo.pdf: 436321 bytes, checksum: 323b66a4d995a23c4570a850ccba61cb (MD5) Previous issue date: 2014-02-27 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The first experiment aimed to evaluate performance and nutritional characteristics of the effect of replacing soybean meal by wheat bran and urea in multiple supplements for beef heifers grazing Brachiaria decumbens Stapf. during the dry season. Forty-five Nelore beef heifers (Age: ±26 months; body weight = 410±2,9) were randomly assigned in five treatments and nine repetitions. The supplements contained approximately 30 % crude protein (CP) altered by replacement of soybean meal to wheat bran and urea in supplements (0, 33, 67 and 100%). The animals of the control treatment were provided only mineral mixture ad libitum and other treatments were provided with 1.0 kg/animal/day of multiple supplement. Supplemented heifers had higher (P<0,10) intakes of dry matter (DM), organic matter (OM), crude protein (CP) and digestible organic matter (DOM) when compared to animals in the control treatment. The replacement of soybean meal for wheat bran and urea influenced in a quadratic manner the intakes of pasture dry matter (PDM), digestible neutral detergent fiber (NDFD), digestible organic matter (DOM) and neutral detergent fiber (NDF) (g/kg BW). The supplementation increased the total tract digestibility of OM and CP, and also increased the levels of DOM (g/kg DM). The level of replacement had negative linear effect on the digestibility of neutral detergent fiber corrected for ash and protein (NDFap), CP and DOM. OM digestibility responded in a quadratic manner the increasing proportions of wheat bran and urea. The different proportions of the ingredients did not affect the microbial efficiency, neither the serum concentration of urea nitrogen, and urinary nitrogen excretion. We conclude that providing multiple supplements improve performance of beef heifers grazing Brachiaria decumbens during the dry season, and that the replacement of soybean meal for wheat bran and urea in the supplement does not compromise the performance of these animals. The second experiment aimed to evaluate nutritional characteristics and productive performance of providing different additives in supplements for Nelore beef heifers grazing Brachiaria decumbens during the dry season. Twenty-four beef heifers (21 months; 383±6 kg) were assigned in a completely randomized design with five treatments and seven repetitions. It was added individually to the supplement three different additives: Exogenous enzymes (Allzyme SSF®), Yeast culture (Yea-Sacc®) and Monensin (Rumensin®). The animals of the control treatment were provided only mineral mixture ad libitum and the other treatments were provided with 1.0 kg/animal/day of multiple supplement (25% CP). The different additives had no effect (P<0.10) on ADG and final body weight (FBW). The animals receiving supplement containing monensin had lower intake of pasture dry matter (PDM, kg) and neutral detergent fiber corrected for ash and protein (NDFap, g/kg BW). There were no positive effects on providing additives in the OM, NDF and CP total tract digestibility when compared to the control group. The microbial efficiency was not influenced by the treatments. We conclude that including exogenous enzymes, yeast culture or monensin individually in multiple supplements does not improve performance of beef heifers grazing brachiaria decumbens during the dry season. / Esta dissertação foi elaborada a partir de dois experimentos com novilhas Nelore submetidas a diferentes estratégias de suplementação em pasto. No primeiro experimento objetivou-se avaliar os efeitos da substituição progressiva do farelo de soja por farelo de trigo e uréia em suplementos múltiplos, sobre o desempenho, o consumo, a digestibilidade, a síntese ruminal de proteína microbiana e o balanço de compostos nitrogenados em novilhas de corte prenhez em pasto de Brachiaria decumbens Stapf no período da seca. Foram utilizadas 45 novilhas Nelore prenhez (5-6 meses de gestação) com idade e peso médio inicial de 26 meses e 410±2,9 kg, respectivamente. O experimento foi conduzido segundo delineamento inteiramente casualizado com cinco tratamentos e nove repetições. Os suplementos continham aproximadamente 30% de proteína bruta (PB) e substituição progressiva do farelo de soja por farelo de trigo e uréia em 33, 67 e 100%. Aos animais do tratamento controle (MM) foi fornecida suplementação mineral ad libitum e aos demais tratamentos foi fornecidos 1,0 kg/animal/dia de suplemento múltiplo. O consumo de matéria seca (MS), matéria orgânica (MO), proteína bruta (PB) e a matéria orgânica digerida (MOD) foram superiores (P<0,10) para os animais que receberam suplementação múltipla em relação aos que receberam suplementação mineral. Observou-se efeito quadrático do nível de substituição de farelo de soja por farelo de trigo e uréia sobre o consumo de MS de pasto (MSP), de FDN digerida (FDNd), de matéria orgânica digerida (MOD) e de FDNcp em g/kg de peso corporal (PC). A suplementação múltipla aumentou os coeficientes de digestibilidade aparente total da MO e da PB e os níveis de matéria orgânica digerida em g/kg de MS (DOM). Houve efeito linear negativo do nível de substituição no suplemento sobre os coeficientes de digestibilidade da FDNcp, da PB e DOM. O nível de substituição influenciou de forma quadrática a digestibilidade da MO. Não foram observados (P<0,10) efeitos dos níveis de substituição no suplemento sobre a síntese microbiana (NMIC), NMIC relativo (NMICR), eficiência de síntese microbiana (EFM) e sobre as concentrações de nitrogênio ureico no soro (NUS) e na urina (NUU). Conclui-se que a suplementação múltipla melhora o desempenho produtivo de novilhas prenhez em pasto de Brachiaria decumbens no período da seca, e que a substituição do farelo de soja por farelo de trigo e uréia em suplementos múltiplos não compromete o desempenho produtivo desses animais. No segundo experimento objetivou-se avaliar o efeito da adição de complexo enzimático (Allzyme SSF®), levedura ativa (Yea-Sacc®) e monensina sódica (Rumensin®) em suplementos múltiplos sobre o desempenho, consumo, digestibilidade, síntese ruminal de proteína microbiana e o balanço de compostos nitrogenados novilhas Nelore em pasto de Brachiaria Decumbens Stapf. no período da seca. Foram utilizadas 35 novilhas de corte Nelore com idade e peso médio inicial de 21 meses e 383±6 kg, respectivamente. O delineamento experimental foi inteiramente casualizado com cinco tratamentos e sete repetições. Os tratamentos foram: suplementação mineral ofertada ad libitum (MM); apenas suplemento múltiplo (S); suplemento mais complexo enzimático (ALLZYME SSF®) (S+E); suplemento mais levedura ativa (Yea-sacc®) (S+L); suplemento mais monensina sódica (Rumensin®) (S+M). Os suplementos múltiplos foram ofertados em 1 kg/animal/ dia e continham aproximadamente 25% de PB. O ganho médio diário (GMD) e também o peso corporal final (PCF) foram superiores (P<0,10) para os animais suplementados, quando comparados aos do tratamento controle (MM). Não houve diferença (P>0,10) para GMD e PCF entre os animais que receberam apenas suplemento (S) e os animais que receberam suplemento aditivado (S+E), (S+L), (S+M). Não foram verificados efeitos positivos da utilização dos aditivos sobre o consumo de nutrientes. A adição de monensina sódica diminuiu o consumo de MSP e de FDNcp em g/kg de peso corporal. Não houve efeito positivo dos aditivos sobre os coeficientes de digestibilidades da MO, da FDN e da PB quando comparados aos animais que receberam suplemento sem aditivo ou apenas mistura mineral. A eficiência microbiana não diferiu entre os tratamentos. Conclui-se que a utilização de complexo enzimático, levedura ativa e monensina sódica adicionados individualmente em suplemento múltiplo, não melhora o desempenho produtivo de novilhas de corte prenhez em pasto de Brachiaria decumbens no período da seca.