The effect of salt-preconditioning Torulaspora delbrueckii cells on fermentation performance

This is an abstract of a paper presented at the 16th European Congress on Biotechnology, Edinburgh, 13-16 July 2014.

Enzymatic hydrolysis and fermentation of ultradispersed wood particles after ultrasonic pretreatment

Background: A study of the correlation between the particle size of lignocellulosic substrates and ultrasound pretreatment on the efficiency of further enzymatic hydrolysis and fermentation to ethanol. Results: Themaximumconcentrations of glucose and, to a lesser extent, di- and trisaccharideswere obtained in a series of experiments with 48-h enzymatic hydrolysis of pine rawmaterials ground at 380–400 rpm for 30min. The highest glucose yield was observed at the end of the hydrolysis with a cellulase dosage of 10 mg of protein (204 ± 21 units CMCase per g of sawdust). The greatest enzymatic hydrolysis efficiency was observed in a sample that combined two-stage grinding at 400 rpm with ultrasonic treatment for 5–10 min at a power of 10 W per kg of sawdust. The glucose yield in this case (35.5 g glucose l−1) increased twofold compared to ground substrate without further preparation. Conclusions: Using a mechanical two-stage grinding of lignocellulosic raw materials with ultrasonication increases the efficiency of subsequent enzymatic hydrolysis and fermentation.

Thermotolerant fermenting yeasts for simultaneous saccharification fermentation of lignocellulosic biomass

Lignocellulosic biomass is the most abundant renewable source of energy that has been widely explored as second-generation biofuel feedstock. Despite more than four decades of research, the process of ethanol production from lignocellulosic (LC) biomass remains economically unfeasible. This is due to the high cost of enzymes, end-product inhibition of enzymes, and the need for cost-intensive inputs associated with a separate hydrolysis and fermentation (SHF) process. Thermotolerant yeast strains that can undergo fermentation at temperatures above 40°C are suitable alternatives for developing the simultaneous saccharification and fermentation (SSF) process to overcome the limitations of SHF. This review describes the various approaches to screen and develop thermotolerant yeasts via genetic and metabolic engineering. The advantages and limitations of SSF at high temperatures are also discussed. A critical insight into the effect of high temperatures on yeast morphology and physiology is also included. This can improve our understanding of the development of thermotolerant yeast amenable to the SSF process to make LC ethanol production commercially viable.

STANDARDIZATION OF CASSAVA MAHEWU FERMENTATION AND ASSESSMENT OF THE EFFECTS OF IRON SOURCES USED FOR FORTIFICATION

Cassava root is the main staple for 70% of the population in Mozambique, particularly in inaccessible rural areas, but is known to be low in iron. Anaemia is a public health problem in mothers and preschool children in Mozambique and up to 40% of these cases are probably due to dietary iron deficiency. The World Health Organization (WHO) and Food and Agriculture Organization of the United Nations (FAO) recognize the fortification of foodstuff as an effective method to remedy dietary deficiencies of micronutrients, including iron. Cassava mahewu, a non-alcoholic fermented beverage is prepared at subsistence level from cassava roots using indigenous procedures. The aim of the study was to standardize mahewu fermentation and investigate if the type of cassava fermented, or the iron compound used for fortification affected the final product. Roots of sweet and bitter varieties of cassava from four districts (Rapale, Meconta, Alto Molocue and Zavala) in Mozambique, were peeled, dried and pounded to prepare flour. Cassava flour was cooked and fermented under controlled conditions (45°C for 24 h). The fermentation period and temperature were set, based on the findings of a pilot study which showed that an end-point pH of about 4.5 was regularly reached after 24 h at 45°C. Cassava mahewu was fortified with ferrous sulfate (FeSO4.7H2O) or ferrous fumarate (C4H2FeO4) at the beginning (time zero) and at the end of fermentation (24 h). The amount of iron added to the mahewu was based on the average of the approved range of iron used for the fortification of maize meal. The mean pH at the endpoint was 4.5, with 0.29% titratable acidity. The pH and acidity were different to those reported in previous studies on maize mahewu, whereas the solid extract of 9.65% was found to be similar. Lactic acid bacteria (LAB) and yeast growth were not significantly different in mahewu fortified with either of the iron compounds. There was no significant difference between cassava mahewu made from bitter or sweet varieties. A standard method for preparation and iron fortification of cassava mahewu was developed. It is recommended that fortification occurs at the end of fermentation when done at household level.

Effect of nitrogen nutrition on yeast ecology and alcoholic fermentation

Mestrado Vinifera Euromaster - Instituto Superior de Agronomia - UL

Ruminal methane emission by dairy cattle in southeast Brazil.

ABSTRACT: Ruminal gases, particularly methane, generated during the fermentative process in rumen, represent a partial loss of feed energy and are also pointed to as an important factors in greenhouse effect. This study aimed at quantifying methane (CH 4) emission rates from lactating and dry cows and heifers, 24 month-old in average, on pasture under Southeast Brazil tropical conditions, using the tracer gas technique, sulphur hexafluoride (SF 6), four animals per category, distributed in four blocks. Measurements were performed in February and June, 2002, with Holstein and Brazilian Dairy Crossbred (Holstein ¾ x Gir (Zebu) ¼), maintained on fertilized Tanzania-grass (Panicum maximum Jacq. cv. Tanzania) and fertilized Brachiaria-grass (Brachiaria decumbens cv. Basilisk) pastures. Heifers of both breeds were maintained on unfertilized Brachiaria-grass to simulate conditions of extensive cattle farming systems. CH 4 and SF 6 levels were measured with gas chromatography. Differences in CH4 emissions were measured (p < 0.05) for genetical groups. Holstein produced more methane (299.3g day?1) than the Crossbred (264.2 g day?1). Lactating cows produced more methane (353.8 g day?1) than dry cows (268.8 g day?1) and heifers (222.6 g day?1). Holstein, with greater milk production potential, produced less CH4 (p < 0.05) per unit of dry matter intake (19.1 g kg?1) than the Crossbred (22.0 g kg?1). Methane emission by heifers grazing fertilized pasture (intensive system) was 222.6 g day?1, greater (p < 0.05) than that of heifers on unfertilized pasture (179.2 g day?1). Methane emission varied as function of animal category and management intensity of production system.

Performance and ruminal morphologic characteristics of Saanen kids fed ground, pelleted or extruded total ration

The aim of the present study was to determine feed intake and average weight gain and to evaluate the ruminal morphologic characteristics of Saanen kids slaughtered at 30, 45 and 60 days of age, according to a completely randomized design. Thirty-six non-castrated male Saanen kids were fed ground total ration, pelleted total ration, or extruded total ration. Feed intake and refusals were controlled daily and the animals were weighed at birth and then once a week. Newborn kids received a milk replacer and were weaned at 45 days. Immediately after slaughter, the animals were eviscerated, the entire digestive apparatus was removed from the carcass. The reticulo-rumen was separated, emptied, washed and weighed. Samples were collected from the dorsal sac, pillar area and ventral sac of the rumen, fixed for about 24h in Bouin's solution, dehydrated, embedded in Histosec and cut into 5 mu m sections. Results showed that dry matter intake (DMI) at weaning and post-weaning and weight gain were higher (P < 0.05) in animals that received the pelleted total ration. The weight of the reticulo-rumen accompanied body development and was heavier in these animals. Histologically, after weaning ruminal papillae were more developed in animals that received pelleted total ration. Length of papillae increased with increase of age. The ratio of papillary height to papillary width increased with age in the ventral sac and until weaning (P > 0.05). We conclude that the pelleting process of the total ration favored increased intake, with a 46.7% increase in weight gain and increase in rumen weight and papillae length, suggesting that best results are obtained with this processing. In general, no difference was observed between the results obtained with extruded and ground total ration, although animals fed extruded total ration showed an increase in rumen weight and papillae width. (c) 2004 Elsevier B.V All rights reserved.

From field to fermentation: characterisation and application of non-dairy cultures in dairy foods

This thesis investigates the phenotypic and genotypic diversity of non-dairy L. lactis strains and their application to dairy fermentations. A bank of non-dairy lactococci were isolated from grass, vegetables and the bovine rumen. Subsequent analysis of these L. lactis strains revealed seven strains to possess cremoris genotypes which did not correlate with their observed phenotypes. Multi-locus sequence typing (MLST) and average nucleotide identity (ANI) highlighted the genetic diversity of lactis and cremoris subspecies. The application of these non-dairy lactococci to cheese production was also assessed. In milk, non-dairy strains formed diverse volatile profiles and selected strains were used as adjuncts in a mini Gouda-type cheese system. Sensory analysis showed non-dairy strains to be strongly associated with the development of off-flavours and bitterness. However, microfluidisation appeared to reduce bitterness. A novel bacteriophage, ɸL47, was isolated using the grass isolate L. lactis ssp. cremoris DPC6860 as a host. The phage, a member of the Siphoviridae, possessed a long tail fiber, previously unseen in dairy lactococcal phages. Genome sequencing revealed ɸL47 to be the largest sequenced lactococcal phage to date and owing to the high % similarity with ɸ949, a second member of the 949 group. Finally, to identify and characterise specific genes which may be important in niche adaptation and for applications to dairy fermentations, comparative genome sequence analysis was performed on L. lactis from corn (DPC6853), the bovine rumen (DPC6853) and grass (DPC6860). This study highlights the contribution of niche specialisation to the intra-species diversity of L. lactis and the adaptation of this organism to different environments. In summary this thesis describes the genetic diversity of L. lactis strains from outside the dairy environment and their potential application in dairy fermentations.

Caracterização bioquímica de proteínas aderidas à fibra do bagaço de cana-de-açúcar obtidas de conteúdo ruminal de ovinos.

A conversão da celulose e da hemicelulose do bagaço de cana-de-açúcar em bioetanol tem utilizado enzimas comerciais importadas, tornando o processo caro, justificando a busca e a produção de enzimas nacionais. O presente trabalho objetivou caracterizar as proteínas aderidas à fibra do bagaço de cana-de-açúcar (BCA), obtidas do conteúdo ruminal de ovinos Morada Nova, quanto à resistência térmica e à variação de pH.