Cassava (Manihot esculenta) is extensively cultivated throughout the tropics and subtropics regions due to its ability to grow in diverse soil conditions and minimal management. Experiments were made to study the cassava root fermentation by yeasts in order to enhance the nutritive value of their products (fresh pulp and chips). Both cassava chip (CC) and fresh cassava root pulp (FCR) samples were fermented by Saccharomyces cerevisiae in solid-liquid media fermentation conditions during 132 hours and dried at 30ºC. Products were analyzed for proximate composition, mineral composition, essential aminoacids and antinutrient content. There were increases (p < 0.01) in protein (30.4% in CC and 13.5% in fermented cassava root -FCR) and fat contents (5.8% in CC and 3.0% in FCR). S. cerevisiae fermented cassava products had very low hydrocyanic acid (HCN) contents (CC, 0.5 mg kg -1 and FCR, 47.3 mg kg -1 ). There was a remarkable increase in lysine content in the fermented cassava chip (FCC). The best acceptability and organoleptic attributes (color, texture and aroma) of enriched cassava chip were achieved after 132h of bioprocessing. The results of this study suggest that FCC can be nutritionally improved with S. cerevisiae for animal feeding. Key words: Saccharomyces cerevisiae, fermentation, cassava chip, fresh cassava root pulp, protein, animal feed ENRIQUECIMENTO DO VALOR NUTRITIVO DA MANDIOCA POR FERMENTAÇÃO COM LEVEDURASRESUMO: A mandioca (Manihot esculenta) é extensivamente cultivada nas regiões tropical e subtropical devido à sua habilidade de crescer em diveresas condições de clima e manejo. Experimentos foram efetuados para estudar o aumento do valor nutritivo de subprodutos derivados de raízes de mandioca (polpa fresca e raspas) por processos de fermentação. Amostras de raspas (RM) e de polpa fresca (PF) foram fermentadas por Saccharomyces cerevisiae, em condições de meio sólido-líquido durante 132 horas e secas a 30ºC. Foram avaliados a composição aproximada, composição mineral, aminoácidos essenciais e conteúdo de antinutrientes dos produtos obtidos. Houve aumentos (p < 0.01) em proteínas (30,4% em RM e 13,5% in PF) e conteúdo de gorduras (5,8% em RM e 3,0% in PF). Os subprodutos de mandioca fermentados por S. cerevisiae apresentaram baixos conteúdos de ácido hidrocianídrico (RM, 0,5 mg kg -1 e PF 47,3 mg kg -1 ). Houve aumento considerável de lisina nas raspas fermentadas (RMF). Valores aceitáveis de cor, textura e aroma das raspas de mandioca enriquecidas formam obtidos após 132 de bioprocessamento. Sugere-se que a RMF pode ser nutricionalmente melhorada para alimentação animal pelo uso de S. cerevisiae. Palavras chave: Saccharomyces cerevisiae, fermentação, raspas de mandioca, polpa fresca de mandioca, proteína, nutrição animal.

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The effect of different tropical feed sources on rumen ecology, cellulolytic bacteria, feed intake and digestibility of beef cattle was investigated. Four fistulated, castrated male crossbred cattle were randomly allocated to a 4×4 Latin square design. The treatments were: T1) urea-treated (5%) rice straw (UTS); T2) cassava hay (CH); T3) fresh cassava foliage (FCF); T4) UTS:FCF (1:1 dry matter basis). Animals were fed concentrates at 0.3% of body weight on a DM basis and their respective diets on an ad libitum basis. The experimental period was 21 days. The results revealed that the use of UTS, CH, FCF and UTS:FCF as roughage sources could provide effective fiber and maintain an optimal range of ruminal pH and NH 3 -N. Total viable and cellulolytic bacterial populations were enhanced (p<0.05) with UTS as the roughage source. Animals fed FCF had a higher rumen propionate production (p<0.05) with a lower cellulolytic bacteria count. Moreover, three predominant cellulolytic bacteria species, namely Fibrobacter succinogenes, Ruminococcus albus and Ruminococcus flavefaciens, were found in all treatment groups. Roughage intake and total DM intake were highest with UTS (2.2 and 2.5% BW, respectively) as the roughage source (p<0.05). Nutrient intake in terms of organic matter intake (OMI) was similar in UTS, CH and UTS:FCF treatments (8.0, 6.8 and 8.7 kg/d, respectively), while crude protein intake (CPI) was enhanced in CH, FCF and UTS:FCF as compared to the UTS treatment (p<0.05). Digestion coefficients of DM and organic matter (OM) were similar among treatments, while the CP digestion coefficients were similar in CH, FCF and UTS:FCF treatments, but were higher (p<0.05) in CH than in UTS. CP and ADF digestible intakes (kg/d) were highest (p<0.05) on the CH and UTS treatments, respectively. It was also observed that feeding FCF as a full-feed resulted in ataxia as well as frequent urination; therefore, FCF should only be fed fresh as part of the feed or be fed wilted. Hence, combined use of FCF and UTS as well as CH and FCF were recommended.

Industrial yeast strains are generally diploid and are often defective in sporulation. Such strains are hence thought to be less tractable for manipulation by genetic engineering. To facilitate more reliable genetic manipulation of the diploid yeast Japanese sake, we constructed variants of this strain that were homozygous for a URA3 deletion, homozygous for either MATa or MATalpha, and homozygous for either the his3 or the lys4 mutation. A ura3-null genotype enabled gene targeting to be undertaken more easily. The TDH3 promoter was inserted upstream of six yeast genes that have been implicated in flavor control to drive their constitutive overexpression. The homozygous MAT alleles, combined with the non-complementary auxotrophic mutations in the targeted transformants, allowed for tetraploid selection through mating. This resulted in the combinatorial construction of tetraploid strains that overexpress two different genes simultaneously. In addition, a recessive mutant gene, sah1-1, that is known to overproduce S-adenosylmethionine, was introduced into the diploid sake strain by the replacement of one wild-type allele and subsequent disruption of the other. The resulting sah1-1/sah1Delta::URA3 strain produced higher amounts of S-adenosylmethionine than the wild type. The novel sake yeast diploid strains we generated in this study can thus undergo simple PCR-mediated gene manipulation and mating in a manner analogous to established laboratory strains. Moreover, none of these sake strains had extraneous sequences, and they are thus suitable for use in commercial applications.

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