Copper is one of the most important trace elements in ruminant nutrition. Its deficiency causes certain pathologies that can be cured by supplementation, by either five ("inorganic") or complexed ("chelated") forms. With the objective of being able to quantitatively compare the distribution of copper between the two forms of supplements, the speciation of copper in ruminant fluids was studied here. For this study, copper lysine supplement was used. It was necessary to first study the acid-base and complexation chemistry of lysine with copper (II). In addition, the complexation chemistry of glycine and histidine, were investigated. Mass balance equations were used to describe the distribution of copper (II) amongst different ligands. The results of the distribution of copper (II) ions in McDougall's solution (a simulated form of bovine saliva), indicate no significant differences in the distribution of copper using the different form of supplements. 1H NMR was used to validate the results from the computer model. Using a combination of the results from the saliva simulation model and the chemical shifts from the NMR studies, the chemical shift changes that would accompany the addition of copper (II) to McDougall's solution were predicted. Results from the models do not show any appreciable differences from experimental values. Rumen samples were collected. Important peaks in the 1H NMR spectrum were assigned. The spectrum indicated that acetic acid, resulting from the fermentation in the rumen, was a good probe for monitoring the speciation pattern. Speciation calculation indicated that the bulk of the copper would be bound to ammonia in the rumen. Changes in chemical shifts that result from the introduction of copper (II) to the rumen contents were predicted. Results were compared with experimental values. Agreement between the two sets of results was found to be satisfactory. The study shows that any advantages that result from the use of copper lysine supplement are not as a result of its remaining intact. Though metal bioavailabilities are hard to predict this approach could help better our understanding of this process. The methods developed here could be extended to other metal complexation problems in biological fluids. (Abstract shortened by UMI.)

Thesis (MScAgric (Animal Sciences))--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Forages are the main feed components in ruminant production systems for the reason that they are often the major source of energy available to the animal. However, only 10 to 35% of energy intake is available as net energy because the digestion of plant cell walls is not complete. This can significantly affect livestock performance and profits in production systems that use forages as a major source of nutrients of the diet. As a result of low and variable nutritive values of forage feedstuffs, attempts to improve ruminal fibre degradability have been an ongoing research topic. The use of exogenous fibrolytic enzymes (EFE) has been proposed as means to improve forage digestibility. Positive results with regard to rumen forage digestibility and other animal production traits have consequently been obtained due to increased rumen microbial activity following EFE addition in ruminant diets. Two EFE (Abo 374 and EFE 2) and one commercial yeast preparation were firstly identified and selected for their potential to improve the cumulative gas production (GP) at 24 hours of a range of feed substrates using the in vitro GP system as a screening step to identify the superior EFE products. The different feed substrates were lucerne hay, wheat straw, wheat straw treated with urea and a commercial concentrate diet. An in vitro experiment was undertaken on these four different substrates in order to evaluate the two EFE and the yeast preparation. This was to identify the most promising EFE capable of producing a significant effect on feed digestibility using organic matter digestibility (in vitro true digestibility) and fermentation characteristics (in vitro GP system). Results from the in vitro evaluation showed that EFE significantly enhanced in vitro DM degradability and GP profiles (P < 0.05). Abo 374 enzyme showed potential to increase in vitro microbial protein synthesis (MPS) of GP residues of the concentrate diet. In addition, no correlation was found between the in vitro MPS and the 48 hours cumulative GP of all the tested substrates (P < 0.05; R2 < 0.30). Treatments were found to increase in vitro MPS, feed degradability and the cumulative GP of different quality forages and the concentrate diet, with Abo 374 being the best treatment (P < 0.05). However in vitro responses of EFE were variable depending on the energy concentration and chemical composition of different substrates. Variation in MPS was mostly due to the low recovery of purine derivates with the purine laboratory analysis. On the basis of these results, Abo 374 was selected and consequently further tested in another in vitro and in situ trial using a mixed substrate of lucerne hay and wheat straw. Abo 374 significantly improved the cumulative GP, in vitro DM and NDF disappearance of the mixed substrate (P < 0.05). In addition, no correlation was found between the in vitro MPS and the cumulative GP at 48 hours (P = 0.68; R2 < 0.25). The in situ disappearance of feed nutrients (DM, NDF and CP) with Abo 374 was similar to the control. The lack of significance of disappearance was probably due to the small number of sheep used in the study and the relatively high coefficient of variation associated with measuring ruminal digestion. Abo 374 significantly increased the in situ MPS (P = 0.0088) of the mixed substrate of lucerne hay and wheat straw. Evidence of the increased MPS and both in vitro and in situ disappearance of DM and NDF resulted from the Abo 374 activity during either the pre-treatment or the digestion process. The addition of Abo 374 to the mixed substrate of lucerne hay and wheat straw appeared to have been beneficial for microbial colonization of feed particles as a result of the increased rumen activity. It could be speculated that the primary microbial colonization was thus initiated, leading to the release of digestion products that attract in return additional bacteria to the site of digestion. This EFE may be efficient to produce some beneficial depolymerisations of the surface structure of the plant material and the hydrolytic capacity of the rumen to improve microbial attachment and the feed digestibility thereafter. Therefore, the mechanism of action by which Abo 374 improved the feed digestion can be attributed to the increased microbial attachment, stimulation of the rumen microbial population and synergistic effects with hydrolases of ruminal micro-organisms. With regard to these findings, the addition of EFE in ruminant systems can improve the ruminal digestion of DM, NDF and CP to subsequently enhance the supply of the metabolizable protein to the small intestine. Key words: crude protein (CP), exogenous fibrolytic enzymes (EFE), dry matter (DM), gas production (GP), neutral detergent fibre (NDF), microbial protein synthesis (MPS). / AFRIKAANSE OPSOMMING: Ruvoere is die hoof-voerkomponent in herkouer produksiesisteme aangesien dit dikwels die vernaamstebron van energie aan herkouer is. Slegs 10 tot 35% van die energie-inname is beskikbaar as netto-enrgie, omdat die vertering van selwande onvolledig is. Dit kan die prestasie en profyt in produksiesisteme drasties beïnvloed waar ruvoere as ’n hoofbron van nutriënte in die dieet gebruik word. Aangesien die nutriëntwaarde van ruvoere laag is en baie varieer, is navorsing vir verbeterde ruminale veselvertering steeds ’n voorgesette onderwerp. Dit is voorgestel dat eksogeniese fibrolitese ensieme (EFE) gebruik kan word vir verbeterde ruvoervertering. Positiewe resultate in ruminale ruvoerverterig en ander diereproduksie-eienskappe, is verkry as gevolg van toenemende rumen mikrobiese aktiwiteit na EFE aanvulling in herkouerdiëte. Twee EFE’s (Abo 374 en EFE 2) en `n gisproduk is geïdentifiseer en geselekteer vir hul potensiaal om die kumulatiewe gasproduksie (GP) na 24 uur met ’n reeks voersubstrate te verbeter met die gebruik van die in vitro GP sisteem as seleksiemetode om die superieure EFE produkte te identifiseer. Die verskillende ruvoersubstrate was lusernhooi, koringstrooi, ureumbehandelde koringstrooi en ’n kommersiële konsentraatdieet. ’n In vitro eksperiment was onderneem om die vier verskillende substrate te gebruik om die twee EFE’s en gisproduk te evalueer. Hierdeur sou die belowendste EFE’s identifiseer kon word wat ’n betekenisvolle effek op ruvoervertering het. Die vertering van ruvoer sal bepaal word deur organiese materiaal vertering (in vitro ware vertering), asook fermentasie-eienskappe (in vitro GP sisteem). Resultate van die in vitro evaluering het getoon dat EFE’s in vitro DM degradering en GP profiele verbeter. Dit blyk dat die Abo 374 ensiem ’n potensiële toemame in in vitro mikrobiese proteïensintese (MPS), soos bepaal deur die GP oorblyfsels van konsentraat diëte, tot gevolg gehad het. Daar was geen korrelasie tussen die in vitro GP en MPS van al die proefsubstrate nie. Dit blyk dat die behandelings ’n toename in in vitro GP, MPS en ruvoerdegradeerbaarheid van lae kwaliteit ruvoer- en konsentraatdiëte gehad het, waar Abo 374 die beste behandeling was. Die in vitro reaksies van die EFE’s was egter wisselend, afhangende van die energiekonsentrasie en die chemiese samestelling van die verskillende substrate. Variasie van MPS was meestal as gevolg van die lae herwinning van purienderivate tydens die purienanalise. Op grond van dié resultate, is Abo 374 geselekteer om verdere toetse in ander in vitro en in situ proewe te doen. Die substraat wat gebruik is, was ’n 1:1 mengsel van lusernhooi en koringstrooi. Abo 374 het die kumulatiewe RP, in vitro DM en NBV verdwyning van die gemengde substraat verbeter. Boonop is geen korrelasie tussen die MPS en in vitro GP gevind nie. In situ verdwyning van DM, NBV en RP was hoër vir Abo 374, maar nie betekenisvol nie. Die gebrek aan betekenisvolle verdwynings mag die gevolg wees van die klein hoeveelheid skape wat in die proef gebruik is, asook die relatiewe hoë koëffisient van variasie wat gepaard gaan met die bepaling van ruminale vertering. Abo 374 het die in situ MPS betekenisvol verhoog. Verhoogde MPS en in vitro en in situ verdwyning van DM en NBV is waargeneemwaarskynlik as gevolg van die aktiwiteit van Abo 374 gedurende die voorafbehandeling óf die verterings proses. Die byvoeging van Abo 374 tot die gemengde substraat van lusernhooi en koringstrooi blyk om voordelig te wees vir mikrobiese kolonisering van voerpartikels as gevolg van ’n toename in rumenaktiwiteit. Die primêre mikrobiese kolonisering het waaarskynlik gelei tot die vrystelling van verteringsprodukte wat addisionele bakterieë na die plek van vertering lok. Die EFE mag geskik wees vir voordelige depolimerisasie op die oppervlakstruktuur van die plantmateriaal, asook verbeterde hidrolitiese kapasiteit van die rumen om sodoende mikrobiese aanhegting, asook ruvoervertering te verbeter. Dus, Abo 374 se meganisme van aksie wat verbeterde ruvoervertering tot gevolg het, kan toegeskryf word aan `n verhoogde mikrobiese aanhegting, stimulering van die rumen mikrobiese populasie en die sinergistiese effek met hidrolases van rumen mikroörganismes. Ten opsigte van die bevindings, kan die byvoeging van EFE in herkouersisteme ruminale vertering van DM, NBV en RP verbeter, wat dan daaropvolgend die dunderm met meer metaboliseerbare proteïn sal voorsien. Sleutelwoorde: eksogene fibrolitiese ensieme (EFE), droëmaterial (DM), ruproteïen (RP), neutraal bestande vesel (NBV), mikrobiese proteïensintese (MPS), gasproduksie (GP).