The aim of this study was to evaluate the effect of glycosylation on exogenous xylanase stability when incubated under proteolytic conditions. Xylanase produced by Trichoderma longibrachiatum, was purified using gel filtration chromatography, ammonium sulfate salt precipitation and dialysis. A partially purified xylanase with Mr of 20- and 10 kDa was identified and contained >65% of the original xylanase activity. Glycoproteins present in the xylanase were identified by thymol sulfuric acid staining or by the FITC-Iabeled lectin method, specific for glycoproteins. This naturally glycosylated xylanase was enzymatically deglycosylated with one of two endo-N-glycosidases: PNGase F or Endo H. Efficiency of deglycosylation was determined with electrophoresis by observing protein mobility shifts or by staining with FITC-Iabeled lectin. The effect of glycosylation on the stability of the exogenous xylanase was tested by incubating the glycosylated or deglycosylated xylanase with rumen fluid (Rf), Prevotella ruminicola culture supernatant (Pr) or a commercial protease from Bacillus subtilis (Bs) for 0, 3, 6, 9 and 24h at 37°C. Results indicated that glycosylated xylanase was significantly more stable (P<0.05) against proteolytic inactivation under the relatively low protease conditions of Rf and Pr (0.018 and 0.046 mg azocasein degraded/ml/h, respectively), but not under high proteolytic conditions of Bs (1.009 mg azocasein/mllh). Also, the glycosylation effect was observed earlier when incubated with the numerous proteases of Rf (3h), than with Pr (9h). These results indicate that glycosylation enhances xylanase stability and therefore is an important characteristic for exogenous enzyme supplements for ruminants. / Dissertation (MSc (Agric))--University of Pretoria, 2005. / Animal and Wildlife Sciences / unrestricted

Thesis (MScAgric) -- Stellenbosch University, 2004. / ENGLISH ABSTRACT: The ruminant has the ability to utilize forages more efficiently than any other production animal. The utilization of forage fibre is an important aspect of ruminant production systems, as this is the main source of energy available to the animal. The availability of high-fibre forage nutrients is, however, restricted by cell wall degradability, and since low quality forages contribute a great deal to ruminant production systems worldwide, the improvement of this degradation process is of major economic importance. The use of exogenous fibre degrading enzymes has been proposed as a means of enhancing this process, with positive results being obtained from in vitro studies incorporating exogenous enzyme preparations. Positive in vivo results with regard to forage digestibility and other animal production parameters have consequently also been obtained following the addition of exogenous fibre-degrading enzyme preparations to the ruminant diet. Two initial screening experiments were undertaken in order to identify fungal enzyme preparations that may have a positive effect on in vitro fibre degradability. The initial screening employed an in vitro organic matter digestibility technique, and was successful in identifying at least six enzyme preparations displaying enhanced digestibility results that were statistically significant. A second in vitro gas production procedure was used to confirm results obtained from organic matter digestibility assays, as well as to increase screening capacity in order to evaluate new enzyme preparations more time-efficiently. Statistical analysis of results obtained from the secondary screening identified various enzyme candidates producing promising results. Only one of these, Abo 374, proved to be statistically superior to the control and other enzyme preparations. A growth trial was subsequently conducted to assess the performance of this enzyme in vivo. The trial involved individual feeding of 32 Dohne Merino ram lambs grouped according to weights into four groups consisting of 8 lambs each. Each group represented a specific application level of enzyme to the wheat straw component of a high fibre diet, amounting to 10, 5, or 1 ml enzyme supematant/kg straw. The enzyme was diluted with water at appropriate rates to obtain an application rate of 300ml/kg straw. The fourth (control) group was treated with water at the same application rate. The trial was conducted over a period of six weeks, during which feed intakes, weekly weight gains, as well as feed conversion efficiencies were recorded. Results suggested significant weight gains in the high (10ml/kg) and medium (5ml/kg) treatment groups, indicated by a P-value of 0.04. Similarly, feed conversion efficiencies were improved for above-mentioned groups (P=0.05), while feed intakes did not differ significantly between the four experimental groups. / AFRIKAANSE OPSOMMING: Die herkouer besit die vermoee om ruvoere beter as enige ander produksiedier te kan benut. Die gebruik van ruvoervesel is 'n belangrike aspek van herkouer produksiesisteme, aangesien ruvoere die hoof bron van energie aan die herkouer verskaf. Die beskikbaarheid van hoe-vesel ruvoer nutriente word egter beperk deur die degradeerbaarheid van die selwand, en aangesien lae kwaliteit ruvoere 'n groot bydrae tot wereldwye herkouer-produksiesisteme maak, is die moontlike verbetering van hierdie degraderingsproses van groot ekonomiese belang. In 'n poging om hierdie verteringsproses te help bevoordeel, is die gebruik van eksogene veselverterende ensieme ondersoek, en positiewe resultate is verkry wanneer hierdie ensieme in in vitro studies gebruik is. Goeie verbeterings ten opsigte van ruvoer verteerbaarheid en ander diereproduksie parameters is ook verkry deur middel van in vivo studies waar eksogene ensieme by die ruvoer van herkouers gevoeg IS. Twee eksperimente is ondemeem in 'n poging om ensiempreparate wat 'n moontlike positiewe effek op in vitro veselvertering mag he, te identifiseer. Die eerste, 'n in vitro organiese materiaal verteerbaarheid tegniek, was suksesvol in die identifisering van minstens ses ensiem preparate wat statisties betekenisvolle verbeterings ten opsigte van verteringsresultate geproduseer het. 'n Tweede in vitro gasproduksie prosedure is vervolgens gebruik om resultate verkry vanaf die eerste tegniek, te bevestig, asook om evalueringskapasiteit te vergroot en sodoende, nuwe ensiempreparate meer tydseffektief te evalueer. Statistiese evaluering van resultate verkry uit die tweede in vitro tegniek het 'n reeks ensieme met positiewe resultate opgelewer. Een van hierdie, Ab0374, het statisties betekenisvolle resultate ten opsigte van die kontrole, sowel as ander ensieme getoon. In 'n volgende eksperiment is 'n groeiproef gedoen om die effektiwiteit van hierdie ensiem in vivo te toets. In die proef is 32 Dohne Merino ramlammers op grond van hul gewig in vier groepe van agt skape elk verdeel, en individueel gevoer. Die groepe het verskillende toedieningsvlakke van die toetsensiem, toegedien tot die koringstrooi komponent van 'n hoe-vesel dieet, ontvang. Toedieningsvlakke was 10, 5, of lml ensiemkonsentraat/kg strooi. Elke groep se ensiemkonsentraat is verdun met die toepaslike hoeveelheid water om 'n toedieningsvlak van 300ml ensiemoplossing/kg koringstrooi te verkry. 'n Vierde groep is behandel slegs met water teen dieselfde toedieningsvlak, en het gedien as 'n kontrole. Die eksperiment is oor 'n periode van 6 weke uitgevoer. Tydens die proeftydperk is voerinnames, weeklikse gewigstoenames, sowel as voeromsetverhoudings, gedokumenteer. Resultate het betekenisvolle gewigstoenames in die hoe (lOml/kg) en medium (5ml/kg) groepe opgelewer, aangedui deur 'n P-waarde van 0.04. Voeromsetverhoudinge het ook verbeteringe getoon vir bogenoemde twee groepe (P=0.05), terwyl voerinnames nie merkbaar tussen die vier groepe verskil het nie.

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).

Copies of the author's previously published articles inserted. Bibliography: leaves 142-159. Provides information on the significance of pea straw as a ruminant feed and the potential of two biological techniques for improving its feeding value.

Attempts to improve upon the crude protein feeding system have strived to characterize feedstuffs with respect to ruminal protein degradability. In vitro an in situ procedures fall short of this goal by not accounting for ruminal turnover. Six lactating cows, fitted with ruminal and duodenal cannulae, were utilized for in vivo determination of protein degradability by employing a double-marker system. Treatments were corn silage-based diets supplemented with 2:1 soybean meal:fish meal nitrogen (SF) or 1:2 soybean meal:fish meal nitrogen (FS) at levels of 15. 9 and 16.0 percent protein. Ruminal cobalt ethylenediaminetetraacetic acid (CoEDTA) infusions marked liquid phase (LP) digesta and Yb-soaked hay was used as a solid phase (SP) marker. Duodenal digesta was separated into SP and LP at 3000xg. Least squares means of LP flows did not differ for total, precipitable protein, or microbial nitrogen, for SF and FS. Total solid phase flow of dry matter was higher for FS (9.06 kg/d) than for SF (7.97 kg/d), although intakes did not differ. Milk composition and yield did not differ for FS and SF. Average daily gain was 0.12 kg/d for SF and 1:26 kg/d for FS. Whole-tract digestibilties were not different although rumen digestibility of dry matter was 24.9% for FS and 35.7% for SF, and duodenal recovery of N was 93.8% for SF and 107.5% for FS. Inclusion of a higher level of fish meal in the diet increased the proportion of feed proteins delivered to the small intestine, increasing weight gain while having no effect on milk production. / Master of Science

Ph. D. / incomplete_metadata

Thesis (MScAgric) -- University of Stellenbosch, 2002. / ENGLISH ABSTRACT: Low quality roughage is often used in the Western Cape as the only source of roughage for the young growing ruminant. Production levels of these animals is unsatisfactory and therefore supplementation is required. It is necessary when small grains are fed to also include protein sources in the diets. For high production, these protein sources must also be protected against rumen degradation. Good results are reported with the treatment of sunflower oilcakemeal (SFOCM) with formaldehyde (0.86 g formaldehyde per 100 g protein). Protecting methionine with 2-methyl maleic anhydride results in an increase in wool production. This study looked at the effect on dry matter intake (DM-intake), wool production and live weight change with supplementing a low quality diet with urea, SFOCM and formaldehyde protected SFOCM (FSFOCM). These diets where also supplemented with no methionine, methionine and 2-methyl maleyl methionine (MMM). The DM-intake of total diet are increased (P<0.05) with the supplementing of oats straw and barley (1.1% of live weight) diet with SFOCM and FSFOCM (949 & 935 vs 848 gis/d). Supplementing MMM decreased (P<0.05) the DM-intake with 9 %. Supplementing FSFOCM increased (P<0.05) live weight change from 19 to 44 gis/d. Wool production are increased (P<0.05) with 38 % with supplementation of MMM. The apparent digestibility of crude protein (CP) of the urea diet are increased (P<0.05) by supplementing methionine when compared with no methionine supplementation (67.28 vs. 71.23 %). The apparent digestibility of crude protein (CP) of the SFOCM diet (63.71 and 64.96 vs. 70.15 %) and FSFOCM diet (66.39 and 66.26 vs. 70.89 %) are increased (P<0.05) by supplementing MMM when compared with no methionine and methionine supplementation. The apparent digestibility of fat of the SFOCM diet (61.01 and 65.65 vs. 48.33 %) and FSFOCM diet (60.93 and 63.83 vs. 38.14 %) are decreased (P<0.05) by supplementing MMM when compared with no methionine and methionine supplementation. The apparent nitrogen balance of the SFOCM diet (-0.15 and 5.41 vs. 4.62 gis/d) and FSFOCM diet (2.93 and 5.60 vs. 6.02 gis/d) are increased (P<0.05) by supplementing MMM when compared with no methionine and methionine supplementation. The DM- and CP-degradability of barley and DM-degradability of oats straw did not differ (P<0.05) between different protein treatments. The higher production with the supplementation of amino acids and proteins can be associated with more amino acids available in the lower digestibility tract and not as a result of better rumen function. / AFRIKAANSE OPSOMMING: Die benutting van strooi as 'n laegraadse ruvoer deur die jong groeiende herkouer is 'n algemene praktyk wat in die Weskaap toegepas word. Produksie is egter laag en aanvulling van beperkende voedingstowwe is dus noodsaaklik. Saam met kleingrane word proteïenbronne meestal as aanvulling gebruik. Vir hoë produksie is dit dikwels nodig om proteïen aan te vul wat teen ruminale afbraak beskerm word. Die formaldehiedbehandeling (0.86 g formaldehied per 100 g proteïen) van sonneblomoliekoekmeel (SBOKM) is alreeds baie suksesvol toegepas vir die beskerming van proteïen teen ruminale afbraak. Deur metionien met dimetielmaleïelsuuranhidried te beskerm, is wolproduksie verhoog. Die studie het die effek van aanvulling van ureum, SBOKM en formaldehiedbehandelde SBOKM (FSBOKM) saam met geen metionien, metionien en 2,3 - dimetielmaleïelmetionien (MMM) op droëmateriaal (DM) -inname, wolproduksie en liggaamsmassaverandering ondersoek. Die DM-inname van jong Merinohamels op 'n hawerstrooi en gars (1.1% van liggaamsmassa) dieet is verhoog (P<0.05) deur die aanvulling van SBOKM en FSBOKM (949 & 935 vs 848 g/s/d). Die DM-inname is egter met 9 % verlaag (P<0.05) deur aanvulling van MMM. Die aanvulling van FSBOKM het die liggaamsmassatoename verhoog (P<0.05) van 19 na 44 gIs/d. Wolproduksie is met 38% (P<0.05) deur MMM-aanvulling verhoog. Die skynbare verteerbaarheid van ruproteïen (RP) by die ureumbehandeling is verhoog (P<0.05) met metionienaanvulling teenoor geen metionienaanvulling (67.28 vs. 71.23%). By SBOKM (63.71 en 64.96 vs. 70.15%) en FSBOKM-behandelings (66.39 en 66.26 vs. 70.89%) is skynbare verteerbaarheid van RP verhoog (P<0.05) met MMM-aanvulling teenoor metionien- of geen-metionienaanvulling. By SBOKM (61.01 en 65.65 vs. 48.33%) en FSBOKM-behandelings (60.93 en 63.83 vs. 38.14%) is skynbare verteerbaarheid van vet verlaag (P<0.05) met MMM-aanvulling teenoor metionien- of geen metionienaanvulling. By SBOKM (-0.15 en 5.41 vs. 4.62 gIs/d) en FSBOKM-bahandelings (2.93 en 5.60 vs. 6.02 gIs/d) is skynbare N-balans verlaag (P<0.05) met MMM-aanvulling teenoor metionien- of geen-metionienaanvulling. Die effektiewe verteerbaarheid van DM en RP van gars en effektiewe verteerbaarheid van DM van strooi is nie beïnvloed (P>0.05) deur die aanvulling van proteïen nie. Die verhoogde produksie kan dus geassosieer word met 'n verhoogde voorsienning van aminosure in die SVK deur aanvulling en nie weens verbetering in die rumenfunksie nie.

Thesis (PhD (Forest and Wood Science))—University of Stellenbosch, 2007. / Sustainable livestock production in semi-arid Botswana could be improved through tree planting on-farm to provide much needed protein and shade. Such action can be encouraged if the growth, productivity, nutritional value of trees and their contribution to mass gain of livestock are known.. A study at Malotwana investigated two indigenous species, Acacia galpinii and Faidherbia albida, and two exotic species, Leucaena diversifolia and L. leucocephala, at three spacings in a randomised complete block design replicated five times. The three spacings represented 400, 317 and 241 trees per hectare. The study was conducted over 6.5 years. Indigenous trees were sampled biennially and exotics annually to evaluate crown width, height, stem diameter, stem number and stem volume index. Complete plant harvesting of exotic trees at 2.5, 4.5 and 6.5 years evaluated agroforestry production. Leaves from all four species and pods of exotic species were analysed for chemical composition. Two groups of eight lambs were balanced for mass at selection for an on-farm feeding trial. Browse from exotic trees, comprising a 2:1 ratio of pods to leaves, was fed at 30% as supplement to hay to one group whose mean mass was contrasted with that of the control group fed sorghum bran at 30%. A. galpinii was a promising species, adaptable to planting away from its origin. Its crown width ranged from 5.86 m in high density plots to 6.08 m at low density and was significantly different among stands (p = 0.0406) at 6.5 years. Diameter at breast height (dbh) was significantly different among stands aged 6.5 years (p = 0.0003) and ranged between 10.38 cm at high density to 12.48 cm in low density plots, demonstrating a capacity to provide both shade and poles on-farm. At 4.5 years, F. albida attained a mean height of 4.5 m and 4.5 cm in dbh but suffered 67% mortality during a severe drought. Annual fodder production of 0.647 and 0.996 metric tonnes ha-1 for leaves and pods of L. diversifolia and 1.237 and 1.431 for L. leucocephala was recorded in years of average rainfall. Yields of 0.3 and 0.59 metric tonnes ha-1 were recorded for both species in the driest year. Equally good agroforestry production was obtained from both low and high density stands suggesting that low density plantings, which foster higher plant survival and reduce disease incidence, are best suited to the semi-arid conditions of Botswana. The crude protein of leaves ranged between 16.26 (L. diversifolia) and 25.25% (F. albida). They were highly digestible with more than twice the calcium content livestock require. Crude protein and digestibility measures were significantly different among leaves and varied significantly at different spacings (p<0.0001). Pods of the exotic species contained significantly more protein than the leaves (p<0.0001). Lambs supplemented with browse gained 102.33 g per animal per day while the control group gained 83.95 g. There were significant differences between groups during growth (p<0.05). Growing of A. galpinii, complemented with L. diversifolia and L. leucocephala, can supply short and long term feed, and greatly enhance livestock production while diversifying farm feed sources

The growing demand for meat worldwide by the increasing human population (6.8 billion) calls for an increase in livestock production as well as attention to environmental sustainability. Production increases are critical especially in Africa with the highest annual population growth rate (2.5%), where most communities rely on livestock for protein supply. Attempts by intensive livestock farming to optimize production are limited by fibrous quality feeds (roughages) and their unavailability in both developed and developing countries. The overall objective of this study was to scan both domestic and wild herbivores in search for microbial ecosystems with superior fibrolytic potential that can be used as feed additives. It was hypothesized that microbes from wild herbivore can improve fibrous feed breakdown in domesticated ruminants. Experiment 1 evaluated the use of fresh or in vitro cultured faecal inoculum (FF) from two Jersey cows as a potential substitute for rumen fluid (RF). Cultured FF was a better substitute for fresh RF as demonstrated by percentage differences in exocellulase activity (0.4%) and true degradability (TD) (7%), compared to the differences observed between fresh RF and FF for exocellulase activity (33%) and TD (14%). It was applied in subsequent experimentation because it was cost effective (no surgery and reduced sample collection time). The second experiment compared the fibrolytic competence of cultured faecal inocula from three hindgut fermenters (miniature horse (mH), horse (H) and Zebra (ZB)) in summer and winter grazing in their natural environment. Both cellulase enzyme assays (exocellulase, endocellulase and hemicellulase) and in vitro maize stover digestibility study ranked the herbivores according to their fibrolytic competence as ZB > H > mH. The effect of cultured faecal inocula from H, ZB and wildebeest (WB) and its combined systems (N1=H+WB, N2=H+ZB, N3=WB+ZB and N4=H+WB+ZB) on the fermentation of maize stover were also evaluated in vitro. Both enzyme assays and MS degradability studies showed that the combined systems were higher (P<0.01) in fibrolytic activities compared to the individual systems. The microbial ecosystems were ranked as N1 > N2 > N4 > H > ZB > WB >N3; and N3 > N1 > N4 > WB > N2 > ZB >H by their exocellulase activity and degradability parameters, repetitively. The diversity of microbial ecosystems was confirmed by numerous active carboxymethyl cellulase bands present on a carboxymethy cellulose zymograms in experiment 4. The combined microbial ecosystems contain more active and variable bands of cellulases than in the individual microbial ecosystems. Systems N3 and N1 were considered as the best inocula for rumen transinoculation studies. Experiment 5 assessed the in vivo effect of direct-fed microbials from N1 and N3 on MS degradation, ruminal fermentation characteristics and cellulase enzyme profile in sheep. Feed dry matter intake increased (P<0.03) in N1 but tended to increase when inoculated with N3. The treatments, N1 and N3 increased (P<0.05) rumen exocellulase (9.4 and 33.2%, respectively) and endocellulase (82.1 and 47.1%, respectively) specific activities but not hemicellulase activity. Maize stover degradability parameters for N3 (TD, degradability of the insoluble fraction of MS, effective degradability, total SCFA and propionate) measured after 96 h of incubation tended (P>0.05) to be numerically different (1.1, 5.4, 7.1 and 7.9%, respectively). Increase in propionate for N3 was accompanied by higher total SCFA and lower CH4. A decrease in CH4 and no difference in CO2 allow both systems to be environmentally friendly since they have been associated with global warming. These studies showed that direct-fed microbials from N1 and N3 inocula have the potential of improving the utilization of maize stover feeds in ruminants, particularly in view of its simplicity and availability which allows it to be implemented at a relatively lower cost compared to other specific strains or microbial cultures. However, more research is required to identify, purify and classify the superior fibrolytic microbes in the most active ecosystems. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The aim of this study was to determine firstly if there exist variations in fibrolysis among herbivore microbial ecosystems and secondly, the effect on fibre hydrolysis of compositing the most active systems with ruminal microbial ecosystem harvested from a Jersey cow. A literature review pointed to the complexity of carbohydrate (fibre) and how the physical and chemical nature of the forage carbohydrate can present barriers that hinder digestion in the rumen, especially its association with hemicelluloses, pectin, lignin and tannins. Fresh rumen fluid was collected from fistulated herbivores (Jersey cow and sheep) and faecal samples from non-fistulated herbivores (buffalo, horse, impala, camel, elephant, llama, sheep, wildebeest and elephant). Crude protein samples were precipitated with 60% ammonium sulfate. Sample activities were monitored and optimised by incubating with carboxymethyl cellulose (CMC) for 2 h at 39°C. The crude protein samples precipitated from the 11 herbivore microbial ecosystems were active. This was confirmed by an increase in enzyme specific activity with a decrease in total crude protein concentration. In vitro pH optimisation showed a broad range of activity for all ecosystems (4.5-8.0) but for the zebra, horse and elephant which peaked at pH 5. In experiment two (Chapter 4), seasonal variation of the enzymes (exocellulase, endocellulase, cellobiase and xylanase) were monitored through winter and summer. Enzyme specific activity of exocellulase, endocellulase, cellobiase and xylanase were determined by incubation with the specific substrates, crystalline cellulose, CMC, pNPG and xylan, respectively. The amount of reducing sugar released was used to determine the enzyme specific activity. Exocellulase analysis was suitable in winter while summer was preferred for carboxymethyl cellulase and xylanase due to their relative abundance. Cellobiase analysis did not depend on any particular season. Eleven herbivore microbial ecosystems were characterised according to their fibrolytic enzyme specific activities. Enzyme catalytic activities were calculated from kinetic parameters (Km and Vmax) obtained from Eisenthal and Cornish-Bowded plots (Chapter 5). Fibrolytic enzyme expression as well as their activities differed among the 11 ecosystems (P<O.OOOI). They were classified into three groups based on fibrolytic enzyme concentrations; group A with high enzyme concentrations (horse, impala, zebra, wildebeest and the elephant), group B with intermediate (cow, llama, camel, buffalo and giraffe) and group C with low enzyme concentrations (sheep). Exocellulase activity was reasonably correlated with endocellulase activity (r = 0.8978). Xylanase activity was also correlated with carboxymethyl cellulase actvity (r = 0.7104). Enzyme kinetic studies revealed that crude protein samples from the horse, zebra, wildebeest and elephant had the highest enzyme catalytic activities. Microbial or enzyme composite systems were created from the most active ecosystems (horse, wildebeest and zebra) in an attempt to improve the Jersey cow system. These systems were B (cow and horse), C (cow and wildebeest), D (cow and zebra) and E (cow, horse, zebra and wildebeest). The specific activities and enzyme efficiencies of these new systems were determined and compared with system A (cow). Microbial synergism of these systems was also investigated by measuring the amount of gas produced and true degradability (TD) after 72 h of incubation. The composite systems Band E were the most active fibrolytic enzyme systems while C and D were intermediate when compared to that of A. In vitro microbial synergism assays showed that systems B, D, and E had the highest potential of improving milky maize stover (MM) and nutral detergent fibre (NDF) fermentation and degradability in Jersey cows. It was concluded that: (i) fibrolytic and hemicellulolytic enzyme concentrations vary from one season to another with the changing forages; (ii) microbial fibrolytic activities vary among animals grazing on the same field or different geographical regions; and (iii) lastly microbial synergisms of active ecosystems have the potential of improving fibre hydrolysis. However, there is a need to conduct in vivo experimentation to determine the real potential of these in vitro observations. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.