Thesis (Ph. D.)--University of Wisconsin--Madison, 1956. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph.D.)--Ohio State University, 2005. / Title from first page of PDF file. Document formatted into pages; contains xvii, 167 p.; also includes graphics. Includes bibliographical references (p. 148-165). Available online via OhioLINK's ETD Center

Thesis (MSc)--Stellenbosch University, 2001. / ENGLISH ABSTRACT: The use of fungal enzymes as ruminant feed digestibility enhancers was investigated. Currently, ruminants may not digest 38 to 80 % of fibrous forages' content. A renewed interest in the potential of feed enzymes for ruminants was prompted by the high costs of livestock production, together with the availability of newer enzyme preparations. Direct application of enzyme preparations can improve in vitro dry matter (DM) and neutral detergent fibre (NDF) degradation, indicating that direct-fed fibrolytic enzymes may be effective in enhancing in vivo digestion of forages. Two commercial enzyme products, Fibrozyme and Celluclast, and fungal extracellular enzyme extracts from Aureobasidium pullulans, Trichoderma reesei, Aspergillus aculeatus, and Thermomyces lanuginosus were evaluated for enhancing in vitro feed digestibility. Fibrozyme addition to both wheat straw and lucerne hay did not improve their in vitro digestibilities, even after a two hour pre-incubation period. The four fungal enzyme extracts did not enhance wheat straw's digestibility, but marginal increases were evident for lucerne hay. Celluclast addition resulted in marginal increases in the digestibility of both oat hay and oat silage, with no enhanced effect on lucerne hay and NaOH-treated wheat straw. No relationship could be found between the level of enzyme activity and the degree of feed digestion in the in vitro assay. Enzyme hydrolysis with Celluclast, in the absence of rumen fluid, gave more conclusive results. All the feed samples tested showed a positive response to Celluclast addition, even the less digestible feeds, namely sugarcane bagasse and wheat straw. In vitro results show that the assays were unsuccessful, because almost all of the experiments conducted showed inconclusive results. Alternative feed evaluation assays, which include the in vivo, in sacco and in situ methods of analysis, as well as gas production measurement and in vitro analysis with the DAISyII system, should be evaluated. A more detailed study of feed digestibility should be motivated by determining which feeds are hydrolysable, their chemical composition, i.e. how accessible the feeds are, and also evaluation of feed mixtures. The enzyme supplements also need to be evaluated for optimum temperature and pH, as well as the compilation of enzyme cocktails. / AFRIKAANSE OPSOMMING: Die gebruik van swamensieme om die verteerbaarheid van herkouervoere te verhoog, is ondersoek. Tussen 38 en 80 % van veselagtige voere se inhoud is tans onverteerbaar. 'n Hernieude belangstelling in die potensiaal van voerensieme vir herkouers word deur die hoë koste van veeproduksie, asook die beskikbaarheid van nuwe ensiempreparate gedryf Direkte byvoeging van ensiempreparate kan die in vitro droëmateriaal (DM) en neutrale onoplosbare vesel (NOV) vertering verbeter, wat daarop dui dat fibrolitiese ensieme wat direk gevoer word, effektief mag wees tydens die in vivo vertering van voer. Twee kommersiële ensiemprodukte, Fibrozyme en Celluclast, en die vier ekstrasellulêre ensieme van vier swamme, naamlik Aureobasidium pullulans, Trichoderma reesei, Aspergillus aculeatus, en Thermomyces lanuginosus is vir hul vermoë om die in vitro verteerbaarheid van voere te verbeter getoets. Byvoeging van Fibrozyme by beide koringstrooi en lusernhooi het geen verbetering in hulonderskeie in vitro verteerbaarheid tot gevolg gehad nie, selfs nie eens na 'n twee uur vooraf inkubasieperiode nie. Koringstrooi se verteerbaarheid is nie verbeter deur die byvoeging van die vier swam-ensiempreparate nie, maar 'n minimale verbetering is wel waargeneem in die verteerbaarheid van lusernhooi. Byvoeging van Celluclast het 'n minimale verbetering in beide hawerhooi en hawerkuilvoer se verteerbaarheid tot gevolg gehad, maar geen effek op lusernhooi of NaOH-behandelde koringstrooi se verteerbaarheid nie. Geen verwantskap is tussen die vlak van ensiemaktiwiteit en die mate van vertering tydens die in vitro toets gevind nie. Ensiematiese afbraak met Celluclast, in die afwesigheid van rumenvloeistof, het meer konkrete resultate gelewer. Al die voermonsters het 'n positiewe respons op die byvoeging van Celluclast getoon, selfs ook die minder verteerbare voere, nl. suikerrietbagasse en koringstrooi. In die wyer konteks was die resulate van die in vitro verteringstoetse egter onbeduidend as gevolg van groot variasie in die metings. Alternatiewe voerontledingstoetse, wat moontlik beter resultate mag lewer, sluit in in vivo, in sacco en in situ analises, asook die meting van gasproduksie en in vitro analise met die DAISyII sisteem. 'n Meer uitgebreide studie van voerverteerbaarheid wat die bepaling van die afbraak van voere, hul chemiese samestelling, met ander woorde toeganklikheid van voere, en die ondersoek van voermengsels behels, behoort aandag te geniet. Die ensiemmengsels behoort ook ten opsigte van samestelling, optimum temperatuur en pH ondersoek teword.

Thesis (MScAgric)--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Two in vitro experiments were conducted to evaluate the effect of exogenous fibrolytic enzyme application on dry matter (DM) and neutral detergent fibre (NDF) degradation and gas production (GP) of mature forages and forage-concentrate mixtures. The forages used in the first experiment were lucerne hay (LH), oat hay (OH) and wheat straw (WS). The same forages were used in the second experiment, but they were mixed with a concentrate feed to make three mixtures consisting of 80% (HC), 50% (MC) or 20% (LC) concentrate. The extracellular enzyme fraction (supernatant) of a fungal strain, ABO 374, was used as feed additive. The supernatant was used in a fresh (SU-ABO374) or lyophilized (CSIR-ABO374) form, the latter being reconstituted with water immediately before application. The liquid supernatants were applied to the incubation medium and not directly to the substrate, at a rate equivalent to 7.5 ml/kg feed DM. In the control treatments of both experiments, water was used instead of the liquid supernatants. For the DM and NDF degradability trials in both experiments, 500 mg forage samples were weighed into 50 x 50 mm dacron bags which were incubated anaerobically at 39ºC in 1.4L of a rumen liquid inoculated buffered medium in 2L fermentation jars. Bags from all treatments were removed after 2, 4, 8, 12, 24, 48, 72 and 96 h of incubation. For the gas production determinations, 500 mg of the respective substrate samples were weighed into 120 ml glass vials which were incubated for 96 h in 40 ml inoculated medium to which 0.5 ml of the respective enzyme solutions were added. Gas pressure was recorded manually with a digital pressure gauge after 2, 4, 8, 12, 24, 48, 72 and 96 h and pressure was converted to volume with a predetermined regression. The 96 h substrate residues were washed, dried, weighed and analyzed for NDF and OM. In both experiments the substrates differed in terms of DM and NDF degradability and gas production rates, but the enzyme treatments had no effect. The lack of response to enzyme application was ascribed to a number of factors, including the fact that enzyme application was into the incubation medium and not directly onto the substrates and also that no significant pre-incubation interaction time was allowed. The same preparations gave positive results in previous trials where they were applied directly onto the substrates and where a pre-incubation interaction time of 16 hours was allowed. (Key words: Exogenous enzymes, forages, concentrate based diets, DM and NDF degradation, gas production ) / AFRIKAANSE OPSOMMING: Die invloed van eksogene fibrolitiese ensieme op in vitro fermentasiekinetika van ruvoer- en gemengde voersubstrate. Twee in vitro-experimente is uitgevoer om die invloed van eksogene fibrolitiese ensieme op droëmateriaal (DM) en neutraal-onoplosbare vesel (NDF) degradering en gasproduksie (GP) van volwasse ruvoersubstrate en ruvoer-kragvoermengsels te bepaal. Ruvoere in die eerste eksperiment was lusernhooi (LH), hawerhooi (HH) en koringstrooi (KS). Dieselfde ruvoere is in die tweede eksperiment gebruik, maar hulle is met ‘n kragvoer gemeng om drie mengsels te maak, bestaande uit 80% (HK), 50% (MK) of 20% (LK) kragvoer. Die ekstrasellulêre ensiemfraksie (supernatant) van ‘n fungiale stam, ABO 374, is as ‘n voertoedieningsmiddel gebruik. Die supernatant is is in ‘n vars (SU-ABO374) of gevriesdroogde (WNNR-ABO374) vorm gebruik, waar laasgenoemde onmiddellik voor toediening gerekonstitueer is. Die vloeistof-supernatante is nie direk op die substrate gevoeg nie, maar tot die inkubasiemedium gevoeg, teen ‘n hoeveelheid ekwivalent aan 7.5 ml/kg voer DM. In die kontrolebehandeling van beide eksperimente, is water in plaas van die vloeistofsupernatante gebruik. Vir die DM- en NDF-degraderingsproewe in beide eksperimente, is 500 mg van die onderskeie ruvoere in 50 x 50 mm dacronsakkies geweeg wat anaerobies by 39ºC geïnkubeer is in 1.4L van ‘n rumenvloeistof-geïnokkuleerde medium in 2L fermentasieflesse. Vir alle behandelings is sakkies na 2, 4, 8, 12, 24, 48, 72 en 96 h inkubasie verwyder. Vir gasproduksiebepalings is 500 mg van die onderskeie substraatmonsters in 120 ml glasbotteltjies geweeg en vir 96 h in 40 ml geïnokkuleerde medium geïnkubeer waarin 0.5 ml van die onderskeie ensiemoplossings gevoeg is. Gasdruk is na 2, 4, 8, 12, 24, 48, 72 en 96 h bepaal met behulp van ‘n digitale drukmeter en druk is met behulp van ‘n voorafbepaalde regressie na volume omgeskakel. Die 96 h substraatresidue is gewas, gedroog, geweeg en ontleed vir NDF en OM. In beide eksperimente het die substrate verskil ten opsigte van DM- en NDF-degradeerbaarheid en gasproduksietempo’s, maar die ensiembehandelings het geen invloed gehad nie. Die gebrek aan respons is aan verskeie faktore toegeskryf, insluitend die feit dat ensiemtoediening in die inkubasiemedium toegedien is en nie direk op die substrate nie, asook die feit dat daar nie ‘n noemenswaardige pre-inkubasie interaksietyd toegalaat is nie. Dieselfde ensiempreparate het positiewe resultate gelewer in vorige proewe waar dit direk op die substraat toegedien is en waar ‘n pre-inkubasie interaksietyd van 16 ure toegelaat is. (Sleutelwoorde: Eksogene ensieme, ruvoere, kragvoerdiëte, DM- en NDF-degradering, gasproduksie)

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

The objective of the study was to evaluate the effect of an exogenous multi-blend enzyme ( -glucanase and xylanase) on the performance of the broiler chickens and laying hens fed diets based on wheat and barley. Experiments were conducted on a flock of broilers and two flocks of laying hens. In both cases feed and water were provided ad libitum. The enzyme effect of enzyme addition on the broiler performance involved 2080 day-old male and female chicks in 48 pens, allocated one of four dietary treatments (0, 50, 100 or 200g/ton enzyme supplementation), to 35 days of age. On day 35, ten birds from each treatment were sacrificed for the analysis of the digestive organs weight (gizzards and livers). The trial was divided into two phases: a starter (1 to 21 d) and grower (22 to 35 d). Feed consumption was measured weekly and birds were also weighed weekly. The investigation of enzyme effect in laying hen diets involved 896 birds for each specific period. Each replicate consisted of four cages (four birds per cage) with a common feeder; 16 hens/pen of 56 pens. Eggs were weighed three times a week, feed consumption weekly and birds every weeks. The addition of a multi-blend enzyme significantly improve body weight, body weight gain, food intake, and feed conversion ratio for both sexes (P<0.05) in broiler chickens. There was a significant improvement in egg production in laying hens (P<0.05). Egg weight and egg mass were not significantly improved. Wheat and barley have cell wall components (arabinoxylans and -glucans respectively) which have a negative effect on the nutritive value of these feeds and therefore performance in poultry fed diets based on these ingredients. Addition of an exogenous multi-blend enzyme( -glucanase and xylanase) could help reduce these effects and improve performance and digestibility values in poultry. The null hypothesis was there will be no difference between supplemented and un-supplemented diets based on wheat and barley in performance of poultry. The results of this study suggest that the inclusion of 50 g/ton enzyme helps improve poultry performance, especially in young birds. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

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.

This research reported here was carried out to examine alternatives to antibiotic growth promoters as a result of their being banned in the animal feed industry. Four experiments were conducted to evaluate the efficacy of non-medicated feed additives as replacements for antibiotic growth promoters in broiler feeds. The additives used were enzymes (a new thermo-tolerant powder enzyme called TXAP, phytase, lipase and a new phytase enzyme derived from E. coli called phyzyme XP), organic acid (Acid Pak), prebiotic (Bio-Mos®) and probiotic (All-Lac XCL). Mashed maize-soya based feeds were used in all the experiments, which were conducted in litter-floor pens. The first experiment was a dose-response trial. Broilers in eight replicate pens of 50 males and 50 females were fed unsupplemented feeds and five additional feeds containing increasing levels of TXAP, from 0.5 to 2.5 g/kg to 42 d. The second experiment used enzyme TXAP with two different enzymes (phytase and lipase), individually or in combination. Six replicate pens of 50 males and 50 females were fed either unsupplemented feeds or one of six additional feeds treated with TXAP, lipase, phytase , a combination of TXAP and lipase, a combination of TXAP and phytase or a combination of all the three enzymes . This trial continued for 42 d. In the third experiment three types of TXAP (Lot 1, 2 and 3) were used, with fixed levels of xylanase and amylase but varying levels of protease activities (4000, 2000 and 1000 U/kg for Lot 1, 2 and 3, respectively) in combination with phyzyme XP for 35 d. The fourth experiment used mannan-oligosaccharide (Bio-Mos®), organic acid (Acid pak 2x), probiotic (All Lac XCL 5x), individually or in combination and an antibiotic growth promoter (Zinc bacitracin) for 42 d. The chickens in this experiment were challenged with Clostridium perfringens (CP) at 21, 22 and 23 d to determine the efficacy of these additives for replacing antibiotics in hindering the effects of CP on the villus surface area. The dose-response trial did not show any significant improvement in broiler performance with any level of inclusion of enzyme TXAP. The results from this study showed some beneficial effects with the use of enzyme TXAP when fed alone and at a young age. Its use when combined with other enzymes and at later stages of growth needs further investigation. Feed additives in experiment 4 prevented the negative effects of CP as the treated chickens did not have lesions on their villus surfaces. The conditions under which these trials were conducted appeared to be such that little benefit was derived from the use of any of the feed additives used. It is possible that under less-hygienic conditions such as those in commercial operations greater benefits from these additives may be realised. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2006.

MSCAGR / Department of Animal Science