Kilian, S. G.Bragg, R. R.Van der Westhuizen, René Johan2016-01-052016-01-05200820082008-11-30http://hdl.handle.net/11660/1969English: The inulin neoseries, trisaccharide, neokestose was produced by the yeast Xanthophyllomyces dendrorhous (Phaffia rhodozyma Y4-3) during growth on sucrose. To produce neokestose, whole cells harvested from the late exponential growth phase were incubated for 36 to 40 h at 25 oC in 0.2 M citratephosphate buffer (pH 7) containing 220 g.l-1 sucrose. Neokestose made up about 50 % of this mixture, which was purified equally well by both a carbon:celite chromatography as well as a batch filtration process, when eluting with similar amounts of water followed by a 50 % ethanol elution step. A final product was combined from various purification runs which consisted of 82.6 % neokestose, 8.7 % sucrose, 7.6 % GF3, 1.2 % glucose and 0.1 % fructose. Lactobacillus and Bifidobacterium genera are considered part of the beneficial group in in the intestine of animal and man. Bifidobacterium levels were higher than Lactobacillus levels in the caeca of New Hampshire layers, whereas in this study only Lactobacillus species were found in broilers. The reason for the absence of the Bifidobacterium species in the caecum of broilers was not determined. The prebiotic effect was evaluated on 5 week old broiler caecal material in vitro over 24 hours based on the viable levels of the total anaerobic bacteria, Lactobacillus and coliforms. The prebiotic effect was also evaluated on viable levels of added Salmonella Typhi, Escherichia coli and Campylobacter jejuni. Volatile fatty acids and pH were measured. The effect of neokestose on these groups was compared to that of inulin, a known prebiotic, and glucose. The total anaerobe and Lactobacillus levels increased over 24 hours for neokestose, inulin and glucose. Although there was no significant difference between the treatments higher levels were found for neokestose and glucose than for inulin. A decrease in the viable levels of E. coli, S. Typhi and C. jejuni were seen over 24 hours. The production of acetic acid, butyric acid and propionic acid was not significantly different for the treatments and the control. The pH decrease over 24 hours for the treatments was significantly different from the control, which indicated that lactate (not measured) production was probably higher in the neokestose, inulin and glucose treatments. In vivo tests are, however, required to fully evaluate the prebiotic and “bifidogenic” effect of neokestose for broilers.Afrikaans: Die trisakkaried neokestose, wat deel uitmaak van die inulien neo-reeks, word geproduseer deur die gis Xanthophyllomyces dendrorhous (Phaffia rhodozyma Y4-3) gedurende groei op sukrose. Selle wat verkry is vanaf die laat eksponensiële groeifase is gebruik vir die produksie van neokestose. Die selle is geïnkubeer by 25 oC vir ‘n periode van 36 tot 40 uur in ‘n 0.2 M sitraat-fosfaat buffer (pH 7) met 220 g.l-1 sukrose. Neokestose het ongeveer 50 % van hierdie mengsel beslaan. Suiwering met geaktiveerde koolstof:celite is ewe goed gedoen deur ‘n kromatografiese asook ‘n “batch” filtrasie proses, mits dieselfde volumes water en 50 % etanol aangewend wod gedurende die elluëringsstappe. Die finale produk wat ‘n samevoeging van verskeie suiwerings was het uit 82.6 % neokestose, 8.7 % sukrose, 7.6 % GF3, 1.2 % glukose en 0.1 % fruktose bestaan. Lactobacillus en Bifidobacterium genera word gesien as deel van die voordelige groep bakterieë wat aangetref word in die ingewandskanaal van mense en diere. Bifidobacterium vlakke was hoër as die Lactobacillus vlakke in die sekums van New Hampshire lê-henne, in vergelyking met braaikuikens waar daar slegs Lactobacillus spesies gevind is. `n Rede vir die afwesigheid van Bifidobacterium spesies in die sekum van braaikuikens kon nie vasgestel word nie. Die prebiotiese effek is geëvalueer op 5 weke oue braaikuikensekummateriaal in vitro oor ‘n tydperk van 24 uur en was gebasseer op die lewensvatbare vlakke van die totale anaërobe bakterië, Lactobacillus en koliforme. Die prebiotiese effek is ook addisioneel geëvalueer in die teenwoordigheid van lewensvatbare Salmonella Typhi, Escherichia coli en Campylobacter jejuni wat toegevoeg is tot die sekummateriaal. Vir addisionelle evaluasie is vlugtige vetsure en pH ook gemeet. Die effek van neokestose op hierdie groepe is vergelyk met die van inulien, ‘n bekende prebiotikum, en glukose. Die totale anaërobiese en Lactobacillus vlakke het toegeneem oor 24 uur vir neokestose, inulien en glukose. Alhoewel daar geen beduidende verskille tussen die behandelings waargeneem is nie, was die vlakke vir beide neokestose en glukose hoër as vir inulien en die kontrole. ‘n Vermindering in die lewensvatbare vlakke van E. coli, Salmonella Typhi en C. jejuni is ook waargeneem oor 24 uur. Die produksie van asynsuur, bottersuur en propioonsuur het nie ‘n beduidende verskil getoon tussen die behandelings en die kontrole nie. Daar was wel `n beduidende verlaging in pH oor 24 uur teenoor die kontrole, wat daarop dui dat laktaat produksie (nie gemeet nie) waarskynlik hoër was in die neokestose, inulien en glucose behandelings. In vivo toetse is egter nodig om die prebiotiese en “bifidogeniese” effek ten volle te evalueer vir neokestose in braaikuikens.enDissertation (M.Sc. (Microbial, Biochemical and Food Biotechnology))--University of the Free State, 2008PrebioticsFunctional foodsChickens -- Feeding and feedsFeed additivesDissertationUniversity of the Free State