Effects of exogenous fibrolytic enzymes on in vitro fermentation kinetics of forage and mixed

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