Dissertations / Theses on the topic 'Fermented food products'

1-4% of pulse fractions including pea protein and fiber, chickpea and lentil flour as well as soy flour and protein concentrate were selected and characterized. As preliminary results the functional properties of pulse ingredients are varied upon their protein content and pH of the food carrier. Orange juice, apple juice, yogurt and two probiotic fermented milk were selected for supplementation. 1% and 2% pulse fractions gave comparable results in terms of turbidity, cloud and visual stability, color and sensory attributes for both orange and apple juices beverages. All supplements improved the acidification rate of yogurt and probiotic cultures, but the highest effects were obtained with probiotic supplementation with lentil and soy flour. As for the main study, skim milk (9.5 % w/v solid content) was supplemented with 1-3% (w/v) lentil flour, pea flour or skim milk powder and they were inoculated with yogurt starter cultures or probiotic (L. rhamnosus). Acid production during the fermentation, the pH, syneresis, color, rheological properties (dynamic oscillation temperature sweep test at 4-50 ˚C), and sensory properties (only for yogurt) were studied after production and 28 days of refrigerated storage.1-3% lentil and pea flour enhanced acid production during yogurt fermentation, but the microbial population (CFU) of both S. thermophilus and L. bulgaricus were in the same range in all lentil and pea flour and skim milk supplemented yogurts, after production. Pea flour supplementation enhanced survival of L. bulgaricus after storage. The pH decreased from 4.5 to 4.1 in lentil flour and from 4.5 to 3.75 in pea flour supplemented yogurts, after 28 days. Syneresis in 1-2% lentil and pea flour supplemented yogurts was higher than other samples. In lentil supplemented yogurts, "a" and "L" values did not significantly differ in all samples and remained constant after 28 days whereas, "b" value increased as a result of supplementation. Pea flour supplementation did not alter redness or greenness of yogurts, but the yellowness was significantly higher than other yogurts. Yogurt with 3% lentil and pea flour had higher storage (G΄) and loss (G˝) moduli in comparison with samples supplemented with 1-3% skim milk and the control yogurt. 1-2% lentil and pea flour supplemented yogurt showed comparable sensory properties in comparison with 1-2% skim milk supplemented and control samples.1-3% lentil and pea flour enhanced acid production during probiotic fermentation, and the CFU's of L. rhamnosus were comparable with non-supplemented control sample after production. After 28 days, the CFU`s of 2% and 3% lentil supplemented probiotic were as high as 1% skim milk supplemented sample and the CFU`s of 3% pea flour supplemented probiotic was the highest followed by 3-2% skim milk and 1-2% pea flour supplemented samples. The pH decreased from 4.50 to 3.90 for lentil flour supplemented probiotics and from 4.50 to 4.04 for pea flour supplemented probiotics, over 28 days. Syneresis in 1-3% lentil and pea flour supplemented probiotic was significantly lower than other samples. All lentil flour supplemented samples had significantly lower "L" values and higher "b" and "a" values in comparison with skim milk supplemented samples. Pea flour supplementation slightly changed the color which was not as light as skim milk supplemented samples and they showed more yellowness in final product after production and storage. Probiotic fermented milk with 1-3% lentil and pea flour showed higher G΄ and G˝ in comparison with other samples.
Des légumineuses tels que des protéines et fibres de pois, farine de pois chiche, de lentille et de soja ont été sélectionnées et caractérisés. Des résultats préliminaires ont montré que des propriétés fonctionnelles ont variés en fonction de la teneur en protéines et du pH des légumineuses employées. Du jus d'orange et de pomme, du yogourt et deux laits fermentés à l'aide de probiotiques ont été supplémentés avec les différentes légumineuses à des taux de 1 à 4%. Les supplémentations à 1 et 2% ont donné des résultats comparables en termes de turbidité, de stabilité, de couleur et d'attributs sensoriels pour les jus d'orange et de pomme. L'addition de légumineuse a permis d'avoir une acidification plus rapide dans les yaourts et les cultures probiotiques, mais le effet le plus important a été obtenu avec farine de lentilles et le soja dans les cultures probiotiques. Comme précédemment, des laits écrémés (9,5% p/v) ont été supplémentés avec 1-3% (p/v) de farine de lentilles, de pois ou de poudre de lait écrémé. Ils ont été inoculés avec des cultures de yogourt, des probiotiques (L.rhamnosus). La production d'acide lors de la fermentation, le pH, la synérèse, la couleur, les propriétés rhéologiques (essai dynamique balayer oscillation de température à 40-50˚C), et les propriétés sensorielles (uniquement pour les yogourts) ont été étudiés après la production et durant 28 jours d'entreposage frigorifique. 1-3% de farine de lentilles ou de pois ont amélioré la production d'acide pendant la fermentation du yogourt, mais les UFC ont les même compte pour les laits suppléments que pour les témoins (lait écrémé). Il est a noter que L. bulgaricus avaient un meilleur taux de survie au jour 28 avec une supplémentation en farine de pois. La diminution du pH dans les yogourts est de 4,5 à 4,1 avec la farine de lentille et de 4,5 à 3,75 avec farine de pois, après 28 jours. La synérèse pour les yogourts supplémentés à 1 et 2% avec de la farine de lentille ou de pois était supérieure d'autres échantillons. Lorsque le taux de supplémentation augmente en farine de lentille ou de pois, il n'y a pas de différence significative pour les valeurs de a alors que la valeur b a augmenté en fonction de la supplémentation.Les yogourts faits de 1 a 3 % farine de lentilles et de pois 1 3% avaient un module élastique (G') et un module visqueux (G˝) plus élevés que les échantillons supplémentés en lait écrémé et que les témoins. Les Yogourts avec 1 à 2% de farine de lentilles et de pois possèdent des propriétés sensorielles comparable a celles des yogourts faits avec 1 a 2% de lait écrémé et celles des témoins. 1-3% de farine de lentilles ou de pois dans des laits avec probiotiques ont amélioré la production d'acide pendant la fermentation, et les UFC de L rhamnosus étaient comparable a ceux des témoins (lait écrémé) après production. Après 28 jours, les UFC pour les échantillons supplémentés avec 2 et 3% de farine de lentille étaient aussi élevées que ceux supplémentés avec 1% de lait écrème et les UFC pour les échantillons supplémentés avec 3% de farine de pois étaient plus élevées que ceux de tous les autres échantillons. Durant les 28 jours de production le pH diminue dans les laits probiotiques contenant de la farine de lentille de 4,50 à 3,90 et pour ceux contenant de la farine de pois de 4,50 à 4,04. La synérèse dans laits probiotiques avec 1 à 3% de farine de lentilles ou de pois a été significativement plus faible que les autres échantillons. Tous les échantillons contenant de farine de lentilles avaient significativement une valeur de L plus bas et des valeurs de b et a plus élevés en comparaison aux échantillons supplémentés en lait écrémé. L'addition de farine de pois a entraîné une modification de couleur b.Les laits probiotiques supplémentés avec 1 a 3 % de farine de lentilles et de pois ont des valeurs de G' et G˝ supérieures aux autres échantillons.

Bakgrund: Med industrialiseringen har de traditionella yoghurtkulturerna med en mångfald av samverkande mjölksyrabakterier fått ge plats åt de mer standardiserade. En gårdskultur framställs genom att obehandlad mjölk spontanfermenteras av gårdens egen bakterieflora och får därigenom en unik karaktär. Bakteriekulturen kan sedan ympas vidare för tillverkning av yoghurt.Syfte: Syftet med studien är framställning och vidareympning samt sensorisk och mikrobiologisk karaktärisering av termofil gårdskultur från obehandlad mjölk. Vidareympningen avser framställning av yoghurt fermenterad av gårdens egen bakterieflora.Metod: Gårdskulturen framställdes och ympades till yoghurt. Yoghurten undersöktes genom mikrobiologisk karaktärisering, antibiotikaresistens och sensorisk profilering samt jämfördes med industriell kultur och en traditionell heirloomkultur.Resultat: Resultatet visade att gårdskulturen skiljde sig både mikrobiologisk och sensoriskt. Gårdskulturen innehöll stammar av enterokocker vilka inte visade på resistens mot analyserade antibiotika.Slutsats: Det är möjligt att framställa en gårdskultur av godtagbar mikrobiologisk och sensorisk kvalitet. Gårdskulturen ger en differentierad mikrobiologisk och sensorisk karaktär i jämförelse med en industriell kultur och en traditionell heirloomkultur Metoden kan vara riskfylld och kulturen bör analyseras med avseende på patogen tillväxt. En unik gårdsyoghurt kan vara en metod för gårdsmejerister att i sitt varumärke bygga på terroir och platsens unicitet.
Background: With industrialization, the traditional yoghurt cultures with a multitude of lactic acid bacteria had to make way for the more standardized. An artisanal farm culture is produced by raw milk spontaneously fermented by the farm's own bacterial flora and thus develops a unique character. The bacterial culture can then be inoculated for the production of yoghurt.Purpose: The pupose of the study is to produce and inoculate as well as sensory and microbiological characterization of a thermophilic artisanal farm culture from raw milk. The inoculation relates to the production of yoghurt fermented by the farm's own bacterial flora.Method: The artisanal farm culture was produced and inoculated into yoghurt which was assessed by microbiological characterization, antibiotic resistance, sensory profiling and then compared with industrial culture and a traditional heirloom culture.Result: The result showed that the artisanal farm culture differed both microbiologically and with regard to sensory paramters. The farm culture contained strains of enterococci which did not show resistance to analyzed antibiotics.Conclusion: It is possible to produce an artisanal farm culture of good microbiological and sensory quality. The artisanal farm culture provides a differentiated microbiological and sensory character in comparison to an industrial culture and a traditional heirloom culture The method may be risky and the culture should be analyzed for pathogens. A unique farm yoghurt can be a method for artisan farm dairies to build their brand based on terroir.

Akamu is a lactic acid bacteria fermented cereal-based food that complements infant diets in most African countries. Uncontrolled fermentation increases the variability in quality and safety of akamu. This study was aimed at the controlled fermentation of akamu with selected lactic acid bacteria (LAB), investigation of the probiotic potential of the LAB and the effect of variation in production method on the product quality and sensory properties. PCR-DGGE analysis of traditional akamu samples revealed LAB community dominated by Lactobacillus fermentum, L. plantarum, L. delbrueckii subsp. bulgaricus and L. helveticus. Isolated yeasts were Candida tropicalis, C. albicans, Clavispora lusitaniae and Saccharomyces paradoxus. The isolated Lactobacillus plantarum strains (NGL5 and NGL7) fermented irradiated ground maize slurries and produced significant levels of lactic acid (>73 mmol L-1) and low pH ≤3.63 displaying inhibitory activity against Salmonella enterica serovar Enteritidis NCTC 5188, Escherichia coli 1077 (NCTC 11560), Bacillus cereus NCIMB 11925, Staphylococcus aureus NCTC 3750 and Listeria monocytogenes NCTC 7973 in MRS agar and E. coli 1077 in maize slurry fermentation. Viability of both strains of L. plantarum at pH 2 after 3 h was reduced from ≥8.26±0.05 to ≤4.94±0.49 Log10 CFU mL-1 while incubation in 0.3% bile allowed growth to 5.73±0.13 and 7.93±0.12 Log10 CFU mL-1 after 6 h for NGL5 and NGL7 respectively. Auto-aggregation of the L. plantarum strains at 37oC (≥25 after 5 h) correlated with adhesion to hydrocarbons (<15, 26, 33 and 64% for Hexane, Hexadecane, Ethyl acetate and Chloroform respectively). The strains failed to exhibit gelatinase or haemolytic activity but adhered to porcine mucin (OD403 nm ≥0.63 with viability ≥6.52 Log10 CFU mL-1) and Caco-2 cells (≥5.13 Log10 CFU mL-1). The ash, mineral (Ca, K, Mg, Na, S and Zn), IDF, SDFP and TDF content of the L. plantarum fermented ground maize slurries were significantly (p≤0.05) higher than that of the traditional akamu but the peak and final viscosities (139.5 and 68.5 cP respectively) were significantly (p≤0.05) the least. The aroma, appearance, colour, flavour and texture of the resultant porridges were liked moderately by 75% of the assessors. This study demonstrated that fermentation with the L. plantarum strains would contribute towards product safety and the L. plantarum strains possessed some probiotic potential that could be beneficial to the consumers particularly in those developing countries were the main staple foods are fermented cereals.

L'objectif de ce travail était de sélectionner de manière rationnelle une bactérie probiotique par la mise en place d'un crible ainsi que d'étudier et de comprendre l'impact du potentiel d'oxydoréduction (Eh) et du bullage de gaz sur la survie de Bifidobacterium bifidum dans un produit laitier fermenté. Nous avons tout d'abord développé des techniques de sélection des bactéries sur des critères de viabilité / vitalité (analyse par cytométrie en flux) ainsi que sur des critères de fonctionnalité (pouvoir antioxydant). Nous avons pu sélectionner des souches d'intérêt industriel ainsi qu'une souche d'étude académique, Bifidobacterium bifidum.Nous avons ensuite étudié l'effet de la modification du Eh par bullage de gaz sur la survie de B. bifidum dans un produit laitier fermenté. Les laits fermentés conditionnés sous atmosphère anaérobie (Azote) et/ou réductrice (Azote-Hydrogène) permettent une meilleure survie de la souche au cours du stockage (28 jours - 4 °C) par rapport au Contrôle. Puis, nous avons analysé l'effet d'une croissance sous différents Eh sur la viabilité en milieu modèle et la fonctionnalité de B. bifidum. Une croissance en condition anaérobie et/ou réductrice permet d'améliorer à la fois la résistance aux stress (stress oxydant d'ordre physiologique, stress aux sels biliaires et stress côlon), le pouvoir réducteur (sels de tétrazolium), le pouvoir antioxydant (test KRL et test des comètes) et l'adhérence par rapport au Contrôle. Une modulation des propriétés biochimiques membranaires sous Azote et sous Azote-Hydrogène pourraient expliquer ces phénomènes. L'augmentation des composés thiols exofaciaux et l'augmentation des acides gras insaturés à longues chaines est observée pour des cellules produites sous conditions Azote et Azote-Hydrogène. Ainsi ces conditions de croissance présentent une amélioration majeure de l'effet probiotique de B. bifidum, à la fois d'un point de vue de sa résistance aux stress que d'un point de vue de sa fonctionnalité

Thesis (MScFoodSc)--Stellenbosch University, 2001.
ENGLISH ABSTRACT: The preservation of various fresh fish products is achieved by either smoking, salting, canning, freezing or fermenting a highly perishable raw product. Since many of these facilities are not readily available, the use of fermentation as a means of preserving the product has been extensively practiced. However, the fermentation of fish is a time consuming practise and only by accelerating the process would it be possible to ensure the production of a more cost effective and readily available safe end-product. The quality of the fermented fish product is partially determined by the fermentation conditions and the metabolic activity of the microbes present. The rapid identification of the microbes present during the fermentation would enable the selection of possible starters to ensure an accelerated production of high quality fermented fish products. This study was thus undertaken to develop identification fingerprints for bacteria isolated from fermented fish products. A 1300 bp fragment of the 16S rRNA genes of each of the bacteria previously isolated was successfully amplified using the PCR technique. The isolates included strains of the genera Bacillus, Staphylococcus, Sphingomonas, Kocuria, Brevibacillus, Cryseomonas, Vibrio, Stenotrophomonas and Agrobacterium. The data obtained can, therefore, be used in the identification of these microbes isolated from other similar fermented fish products. The fingerprints could also be used to assist in determining the dominant microbial populations responsible for the characteristic qualitative changes occurring in the fish product during fermentation. The microbial composition of a fermenting fish product partially determines the quality of the end-product, therefore, the use of selected bacterial starters could result in the accelerated production of a microbial safe fermented fish product. A further objective of this study was to accelerate the production of a fermented fish product by inoculating macerated trout with either selected lactic acid bacteria (LAB) or with selected bacteria with high proteolytic activity over a 30 day fermentation period. The LAB included a combination of Lactobacillus plantarum, Lactococcus diacetylactis and Pediococcus cerevisiae strains, whereas the bacteria with high proteolytic activity included strains of Kocuria varians, Bacillus subtilis, two strains of B. amyloliquefaciens and a combination of these bacterial species. The quality of the fermented product was determined using changes in product pH, titratable acidity (%TA) and free amino nitrogen (FAN) formation as efficiency parameters. The data obtained during the fermentation of the macerated trout showed that the selected starters did not have a significant effect on the pH decrease in the product over a 30 day fermentation period. The LAB strains did not have a significant effect on the %TA of the fermenting fish product, yet the presence of these bacteria appeared to limit the FAN production in the product. The bacteria with high proteolytic activity resulted in slightly enhanced %TA values and a higher FAN content in the fermented product. It was also determined that the LAB and Kocuria varians, in contrast to the Bacillus spp. inoculums, did not survive the fermentation conditions well, possibly due to the low pH environment. The presence of the starter bacteria in the fermenting fish mixture at the end of the fermentation was also successfully determined with the use of the PCR-RFLP technique. The fermented fish product, obtained at the end of the fermentation period, had a good aroma and compared favourably to similar commercially available fermented fish products. The use of different microbial starters could in future enable the production of a diverse range of high quality products, which could be produced and marketed locally.
AFRIKAANSE OPSOMMING: Die preservering van ‘n verskeidenheid vars vis produkte word bereik deur die hoogs bederfbare produk te rook, te sout, te blik, te vries of te fermenteer. Aangesien baie van hierdie fasiliteite nie geredelik beskikbaar is nie, is die gebruik van fermentasie as ‘n preserverings metode al ekstensief beoefen. Die fermentasie van vis is egter 'n tydsame proses en slegs deur die versnelling van die proses sal dit moontlik wees om die produksie van ‘n meer koste effektiewe en geredelike beskikbare veilige eindproduk te verseker. Die kwaliteit van die gefermenteerde vis produk word gedeeltelik bepaal deur die fermentasie kondisies en die metaboliese aktiwiteit van die mikrobes teenwoordig. Die vinnige identifikasie van die mikrobes teenwoordig gedurende die fermentasie sal die seleksie van moontlike suursels om die versnelde produksie van hoë kwaliteit gefermenteerde vis produkte moontlik maak. Hierdie studie is dus onderneem om identifikasie vingerafdrukke vir bakteriee wat gei'soleer is van gefermenteerde vis produkte moontlik te maak. ‘n 1300 bp fragment van die 16S rRNA gene van elkeen van die bakteriee wat voorheen gei'soleer is, is suksesvol geamplifiseer deur die PCR tegniek. Die isolate sluit in stamme van die genera Bacillus, Staphylococcus, Sphingomonas, Kocuria, Brevibacillus, Cryseomonas, Vibrio, Stenotrophomonas en Agrobacterium. Die data kan dus gebruik word in die identifikasie van hierdie mikrobes as dit gei'soleer word van ander gefermenteerde vis produkte. Die vingerafdrukke kan ook gebruik word om die dominante mikrobiese populasies wat verantwoordelik is vir die kenmerklike kwalitatiewe veranderinge wat plaasvind in die vis produk gedurende die fermentasie, te identifiseer. Die mikrobiese samestelling van ‘n fermenterende vis produk bepaal gedeeltelik die kwaliteit van die eindproduk, daarom kan die gebruik van geselekteerde bakteriese suursels die versnelde produksie van ‘n mikrobies veilige gefermenteerde vis produk teweeg bring. ‘n Verdere doel van hierdie studie was om die produksie van ‘n gefermenteerde vis produk te versnel deur fyngemaakte forel met of geselekteerde melksuurbakteriee of met geselekteerde bakteriee met hoë proteolitiese aktiwiteit te inokuleer oor ‘n 30 dag fermentasie periode. Die melksuurbakteriee het ingesluit ‘n kombinasie van Lactobacillus plantarum, Lactococcus diacetylactis en Pediococcus cerevisiae, terwyl die bakterieë met hoë proteolitiese aktiwiteit stamme van Kocuria varians, Bacillus subtilis, twee stamme van Bacillus amyloliquefaciens en ‘n kombinasie van hierdie bakteriese stamme ingesluit het. Die kwaliteit van die gefermenteerde produk is bepaal deur die veranderinge in die pH, titreerbare suur (%TS) en vrye amino stikstof (VAS) vorming van die produk as effektiwiteits parameters te gebruik. Die data wat verkry is gedurende die fermentasie van die fyngemaakte forel het gedui daarop dat die geselekteerde suursels nie ‘n merkwaardige effek op die afname in pH in die produk oor ‘n 30 dag fermentasie periode het nie. Die melksuurbakteriee het nie ‘n merkwaardige effek op die %TS van die gefermenteerde vis produk gehad nie, terwyl dit geblyk het dat die teenwoordigheid van hierdie bakterieë die produksie van VAS in die produk belemmer het. Die bakteriee met hoe proteolitiese aktiwiteit het ‘n effense verhoogde %TS en ‘n hoër VAS inhoud in die gefermenteerde produk veroorsaak. Dit is ook bepaal dat die melksuurbakteriee en Kocuria varians, in teenstelling met die Bacillus spp. inokulums, nie die fermentasie kondisies goed oorleef het nie, moontlik as gevolg van die lae pH omgewing. Die teenwoordigheid van die suursel bakteriee in die fermenterende vis mengsel aan die einde van die fermentasie is ook suksesvol bepaal met die PKR-RFLP tegniek. Die gefermenteerde vis produk, verkry aan die einde van die fermentasie periode, het ‘n goeie aroma gehad en het goed vergelyk met soortgelyke kommersieel beskikbare gefermenteerde vis produkte. Die gebruik van verskillende mikrobiese suursels kan in die toekoms die produksie van ‘n diverse reeks hoë kwaliteit produkte wat plaaslik geproduseer en bemark kan word moontlik maak.

Ogi is a fermented food made from maize, sorghum or millet which serves as complementary food for infants and breakfast for adults in Nigeria, West Africa. This study characterized the microbial diversity of maize and sorghum grains and ogi produced by their natural fermentation in an attempt to understand the roles of the key microbial species and the impact of the population dynamics and selected species on changes in nutritional composition and aroma notes of ogi during fermentation. A combined approach of culture dependent and culture independent methods of analysis was applied to investigate the microbial community of grains and ogi from two different sources. Microbial diversity and viable populations varied with the source of the grain. Bacterial and fungal genera identified with the partial 16S rRNA and 26S rRNA sequence analysis respectively in maize and sorghum were Bacillus, Enterobacter, Micrococcus, Kytococcus, Pantoea, Staphylococcus, Amycolatopsis, Methanoculleus, Aspergillus, Penicillium, Eupenicillium, Acremonium, Schizosaccharomyces, Meyerozyma, Hyphopichia, and Pichia in maize grains; Enterococcus, Enterobacter, Pantoea, Bifidobacterium, Aspergillus, Cladosporum, and Penicillium in maize ogi; Enterococcus, Enterobacter, Pantoea, Aeribacillus, Cyanobacterium, Acinetobacter, Fusarium and Trametes in sorghum grains; and Pediococcus, Lactobacillus, Enterococcus, Bacillus, Cladosporum and Penicillium in sorghum ogi. Similar species were observed in both sources of maize while those of sorghum differed slightly. Predominant microbes included species of Enterobacteriaceae and moulds. Acetic acid bacteria were not identified as part of the diverse community. Following the predominance of moulds during the natural fermentation, preliminary screening was performed by PCR using specific biosynthetic gene primers to test whether they are the mycotoxin producing species. None of the genes tested were detected by PCR thus they may not be the toxin producing species. Starch, non-starch polysaccharide (NSP), phytate and phenolic compounds were determined in the grains and respective ogi to ascertain the levels of these nutritionally important components in the naturally fermented ogi and the impact of the varying microbial populations on the fate of these compounds during fermentation. In the grains, the average starch and NSP contents in each case were 80.35 g/100g and 9.40g/100g in maize and 93.12 g/100g and 8.14 g/100g in sorghum. Out of the total in grain the average percentage recovery of starch and NSP respectively in the ogi showed 63% and 42% in maize and 58% and 27% in sorghum. Maize showed good starch and fibre (NSP) retention than sorghum after fermentation. To further understand the types and levels of polymers in NSP hydrolysis in ogi fermentation, HPLC analysis of the hydrolysed extract was performed. Glucose was entirely present in maize and sorghum ogi which represents the beta-D-glucans while arabinose and xylose (in maize only), mostly lost with the pomace, signify the arabinoxylans. Overall variations in the microbial populations of sorghum seemed causal to the difference in starch and NSP recoveries. Phytate was assessed based on release of total phosphorus in the samples by enzymatic and chemical methods. Recovery of phytate in the naturally fermented ogi ranged from 18-25% in maize and 40-48% in sorghum suggesting greater phytase activity and more nutrient bioavailability in maize ogi than in the sorghum. Greater activity in maize reflects the presence of phytate hydrolysing species such as Aspergillus in the grain. Total phenolic content (TPC) was assessed by Folin-Ciocalteu colorimetric method after direct extraction of samples by saponification. TPC in the original grains ranged from 410–437 mg GAE/100g in maize and 221–247 mg GAE/100g in sorghum. Due to the nutritional significance, the amount of phenolics that are either freely soluble or are covalently bound to the food matrix were assessed. Soluble phenolics in ogi ranged from 16-38% in maize and 32-49% in sorghum based on the total soluble fraction in the original grain. In all cases loss of soluble phenolics with the waste waters accounted for 12-25% and 31-39% with the pomace. Only the LAB population seemed to correlate with the release of phenolics in the natural fermentation. Given the higher value of soluble phenolics, naturally fermented sorghum ogi appeared to have higher antioxidant potential than the maize ogi. Furthermore an attempt was made to ascertain whether the use of selected microbes would improve the antioxidant properties and aroma of ogi while minimizing the incidence of pathogens due to chance inoculation. Thus the impact of selected LAB (Pediococcus pentosaceus) and fungi (T. hirsuta and A. zeae previously shown to have phytase activity) on changes in phytate, phenolics and aroma of ogi was assessed following a parallel experiment to the previous study but using autoclaved grains. Five fermentation treatments of the pure and co-cultures were investigated. Cell populations in all culture fermentations varied and reached the average maximum of log 6-9 cfu/ml. Changes in the distribution of bound and soluble phenolics were observed showing esterase activity. Leaching of phenolics was evident in all cases but was higher in the sorghum fermentations. Higher levels of soluble phenolics were recovered in pure culture fermented ogi using T. hirsuta or P. pentosaceus than in the natural fermentation having 76% and 45% of the original soluble fraction in maize and sorghum respectively. This suggests greater antioxidant potentials than the naturally fermented ogi. Pure culture fermentations using T. hirsuta and co-culture of P. pentosaceus with A. zeae reduced phytate by 97% and 96% in maize and sorghum ogi respectively showing greater phytase activity and more nutrient bioavailabilty in the ogi than in the natural fermentation. The aroma profile of ogi was analysed by solid-phase microextraction and gas chromatography-mass spectrophotometry (SPME GC-MS). Ethyl acetate, butyl acetate and ethyl hexanoate were observed as the key active aroma components in ogi. The ester, methyl thiobutanoate was found to be unique to the naturally fermented ogi suggesting that it may have been generated by species other than the selected starter organisms. Overall in both natural and starter culture fermentations, maize ogi showed high relative abundance of volatile components suggesting good substrate compatibility and utilization during fermentation. Thus compounds with high threshold values may be significant in the aroma notes of maize ogi. P. pentosaceus and T. hirsuta in pure and in co-culture fermentations produced ogi with aroma notes mostly related to the naturally fermented product. In conclusion the diversity and levels of the initial microflora and the structural composition of grain could be major factors contributing to the nutritional compositional changes in ogi fermentation.

Les folates sont des vitamines indispensables à tous les âges, particulièrement pendant la grossesse et l’enfance, étant donné leur fonction dans la division cellulaire. Le régime alimentaire en Afrique est essentiellement basé sur les céréales, consommées toujours après transformation. La fermentation est l’un des moyens de transformation des céréales pouvant augmenter les folates dans les aliments. L’objectif de ce travail était d’utiliser la fermentation pour augmenter les ingérés en folates des populations africaines via la consommation d’aliments céréaliers fermentés. Sept aliments céréaliers fermentés couramment consommés en Afrique de l’Ouest ont été investigués. La plupart des aliments avaient des teneurs en folates (1,8–31,3 µg/100g matière fraîche) inférieures à celles des céréales de départ (13,8-73,4 µg/100g matière fraîche). Des pertes en folates ont lieu au cours de certaines étapes de procédés dont le décorticage, le séchage, le trempage, le broyage et la filtration. Toutefois, la fermentation a permis une augmentation de la teneur en folates dans certains aliments. La bioaccessibilité des folates, évaluée à l'aide d'un modèle de digestion statique in vitro, variait de 23% à 81%. Elle était influencée par la matrice alimentaire et la stabilité des folates au cours de la digestion. Il a été calculé qu’au maximum 8% des besoins journaliers en folates des jeunes enfants consommant l’un des aliments étudiés pourraient être couverts. Des essais d’inoculation de bouillies fermentés à base de mil avec des souches de bactéries lactiques sélectionnées pour leurs propriétés nutritionnelles (synthèse de folates, hydrolyse de l’amidon) permettaient d’augmenter significativement les teneurs en folates (jusqu’à 8,7 µg/100 g matière fraîche) par rapport à leur équivalent préparées de manière traditionnelle (2,5-5,4 µg/100 g matière fraîche). L’inoculation par un pied-de-cuve provenant d’une fermentation spontanée permettait aussi une augmentation significative des teneurs folates (jusqu’à 7,4 µg/100 g matière fraîche). La caractérisation de la diversité bactérienne de 7 aliments céréaliers fermentés du Burkina Faso, d’Ethiopie et de la Finlande montrait que les bactéries lactiques du genre Lactobacillus, Enterococcus, Weissella, Pediococcus, Lactococcus et Streptococcus étaient les principaux acteurs de la fermentation de ces aliments. Toutefois, une présence non négligeable d’autres microorganismes potentiellement pathogènes des genres Bacillus, Pseudomonas, Staphylococcus, Erwinia, Klebsiella, Escherichia et Acinetobacter a été identifiée. Cette contamination était liée à certaines étapes du procédé de transformation des céréales dont le stockage et broyage dans les moulins publics. Ces microorganismes pathogènes étaient réduits par fermentation et finalement éliminés après l'étape de cuisson
Folates represent an essential vitamin in the human diet at all ages, particularly during pregnancy and infancy, as it is required for the production of new cells. In many African countries, the main staple foods are based on cereals, which are always consumed after processing. Fermentation is one of the processing, which could increase folate contents in foods. The objective of this work was to increase folates intake of African people through the consumption of cereal-based fermented foods using fermentation. Seven types of cereal-based fermented foods (CBFF), commonly consumed in West Africa, were investigated in this study. Total folate content of cereal-based fermented ranged between 1.8 and 31.3 µg/100g fresh weight, and was almost always lower than in the raw material (13.8-73.4 µg/100g fresh weight). Folate losses occurred during some processing steps like debranning, soaking and drying steps. However, fermentation was able to increase the folate content in some CBFF. Folate bioaccessibility was assessed using a static in vitro digestion model, and ranged from 23% to 81%. The bioaccessible folate content was influenced by total folate content, the structure of food matrices that modulated folate release, and folate stability during digestion process. Calculations of the contributions of CBFF to the reference nutrient intake for folate showed that folate intakes from these foods would cover a maximum of 8% of the folate requirements for young children. Porridges prepared with starter cultures of lactic acid bacteria selected for the nutritional properties (folate synthesis, starch hydrolysis) had significantly higher folate contents (up to 8.7 µg/100 g fresh matter) than the porridge prepared using the traditional process (2.5-5.4 µg/100 g fresh matter). Back slopping using an inoculum from a spontaneous fermentation also enabled an interesting increase in folate contents (up to 7.4 µg/100 g fresh matter). The bacterial diversity of seven cereal-based fermented foods from Burkina Faso, Ethiopia and Finland were assessed using 16S rRNA amplicon sequencing. Lactic acid bacteria genus, including Lactobacillus, Enterococcus, Weissella, Pediococcus, Lactococcus and Streptococcus were the main bacteria present in cereal-based fermented foods. Several potentially pathogenic bacteria, namely, Bacillus, Pseudomonas, Staphylococcus, Erwinia, Escherichia, Klebsiella and Acinetobacter were also found in some intermediary products resulting from storage and wet milling. These microorganisms were reduced by fermentation and finally removed after the cooking step

Thesis (PhD Food Sc)--Stellenbosch University, 2012.
ENGLISH ABSTRACT: The fermentation of milk has been known for millennia and leads to nutritious and prolonged shelf-life dairy products. In Southern Africa, traditional fermented dairy products have the same value as local staple foods and are consumed as a part of or as a whole meal. However, the retail price and the technology make many commercialised fermented dairy products unaffordable to the majority of the population. There is thus a need for a healthy nutritious low-cost easily prepared fermented dairy product. A product that could be the answer to the above need, is Kefir. The principle advantage is that the Kefir beverage is made from reusable Kefir grains, which unfortunately are not easily available and grow slowly. Kefir grains can only be obtained from pre-existing grains, which presents a problem in the marketing of the grains. A mass culturing technique was developed to produce large masses of grains but preparation of Kefir using these grains results in a product (MG Kefir) lacking in the sensory attributes of Traditional Kefir. Thus, the overall objective of this research was to determine the impact of environmental factors on the metabolic profiles of Kefir produced using different Kefir grains and this was then followed by the subsequent enrichment of Kefir prepared with mass cultured grains so as to obtain a Kefir beverage that has improved organoleptic qualities. To determine the impact of environmental factors Traditional and MG Kefir were prepared under controlled and uncontrolled conditions. Traditional Kefir was found to give the best beverage and was thus considered as the control. Under controlled conditions the optimum incubation temperature for the production of Kefir was 22ºC as over-acidification was observed at 25ºC. The metabolic profiles of both Traditional and MG Kefir showed that both contained acetaldehyde, ethanol, acetone, diacetyl and acetic acid. In addition, the metabolic profiles revealed that an inadequate ratio of diacetyl to acetaldehyde as well as the lack of ethyl acetate was responsible for the flavour defect in MG Kefir. In order to overcome this defect, citrate and ascorbate (0.015 % w.v-1) were added during Kefir fermentation to enhance the diacetyl and ethyl acetate production. This addition showed a positive impact on diacetyl but not on ethyl acetate production. Improvement of the overall flavour of Kefir was observed as the ratios of diacetyl to acetaldehyde were higher (0.21 – 0.5) in the samples with added citrate and ascorbate than in the samples without (0.12 – 0.17). The production of ethyl acetate in MG Kefir was enhanced by combining the effects of longer incubation (24 h + 18 h at 22ºC), addition of ethanol and acetic acid at 0.79% (m.v-1) and the addition of either Lactococcus lactis ssp. diacetylactis biovar diacetylactis 318 or Candida kefyr 1283. The best yields were obtained in samples containing C. kefyr 1283 and only added ethanol (9.22 mg.L-1), indicating that ethanol is an important factor in ethyl acetate production by Kefir starter strains and suggesting that the absence of ethyl acetate is an indication that the grains do not contain a yeast capable of producing sufficient ethyl acetate. During this investigation, the impact of ethyl acetate on the organoleptic quality of Kefir during storage at refrigerated and room temperatures were also studied. The results indicated that refrigerated Kefir contained up to 40 mg.L-1 of ethyl acetate and was not found defective and thus ethyl acetate was considered a positive contributor to Kefir flavour. This is of particular interest as ethyl acetate is a potent flavour compound at concentrations below 5 mg.L-1. Improvements of MG Kefir’s flavour were successful and will be of value for commercial Kefir production where the main aim is to optimise the flavour of Kefir. However, stabilising the grain microbial consortium was found to be important as it is responsible, over time, for both stable and acceptable Kefir. Acceptability of Traditional, MG and other Kefirs (Candi-Kefir and Lacto-Kefir) prepared with microbially stabilised MG was evaluated by 85 consumers. Results indicated that pH (r = 0.978; p < 0.05) was a significant driver of liking for flavour, especially for female consumers (r = 0.982; p < 0.05). In addition, three clusters, each characterised by different liking attributes were identified. Cluster I generally disliked all the products whereas slight acidic Kefir such as Candi-Kefir (7.63) and Lacto-Kefir (7.09) were preferred by Cluster III. Cluster II showed preference to Kefir with moderate acidity and high ethanol content. In that regard, Traditional Kefir obtained the best score (7.50) and MG Kefir the lowest score (4.87). The sensory study is of value as it led to the identification of the drivers of consumers liking by the different types of consumers. In the course of this project, near infrared reflectance spectroscopy was developed as a rapid method to estimate lactic and acetic acids, which are the organic acids responsible for acidity in Kefir, as well as pH and titratable acidity (TA). The results showed that the calibration models for lactic acid (RPD = 2.57 – 3.16), pH (RPD = 2.90) and TA (RPD = 2.60) were good for screening purposes (2 < RPD < 3); indicating that these models would show if the concentrations of lactic acid, the pH or the TA varied from the normal range. This study has demonstrated that the flavour of MG Kefir, prepared with enriched grains, was successfully improved and has provided some understanding on the preference liking of Kefir, an unknown fermented dairy product to South African consumers.
AFRIKAANSE OPSOMMING: Die fermentering van melk is al vir millennia bekend en lei tot voedsame suiwelprodukte met 'n verlengde raklewe. In Suidelike Afrika het tradisioneel gefermenteerde suiwelprodukte dieselfde waarde as plaaslike stapelvoedsels en word dit as 'n maaltyd of as deel van 'n maaltyd geëet. Die kleinhandelsprys en tegnologie van kommersieel gefermenteerde suiwelprodukte maak hierdie produkte egter onbekostigbaar vir die grootste deel van die populasie. Daar is dus 'n behoefte aan 'n gesonde, voedsame, goedkoop, maklik-om-te-berei gefermenteerde suiwelproduk. 'n Moontlike produk om aan die bogenoemde te voldoen is Kefir. Die hoof voordeel is dat die Kefir drankie van herbruikbare Kefirkorrels gemaak word, maar ongelukkig is hierdie korrels nie vrylik beskikbaar nie, en vermeerder dit stadig. Kefirkorrels kan net van reeds bestaande korrels verkry word wat problematies is vir die bemarking van hierdie korrels. 'n Massakwekingstegniek is ontwikkel vir die produksie van groot hoeveelhede korrels maar die voorbereiding van Kefir met hierdie korrels lei tot 'n produk (MG Kefir) wat sensories minder aanvaarbaar is as tradisionele Kefir. Die hoofdoel van hierdie navorsing was dus om die invloed van omgewingsfaktore op die metaboliese profiele van Kefir, berei deur gebruik te maak van verskillende Kefirkorrels, te bepaal. Dit is gevolg deur die verryking van Kefir berei van massagekweekte korrels om 'n Kefir drankie met verbeterde organoleptiese kwaliteite te verkry. Tradisionele en MG Kefir is voorberei onder gekontroleerde en ongekontroleerde toestande om die impak van omgewingsfaktore te bepaal. Die beste drankie is van tradisionele Kefir verkry en is dus beskou as die kontrole. Die optimum temperatuur vir die produksie van Kefir onder gekonroleerde toestande is 22ºC aangesien oor-versuring by 25ºC waargeneem is. Die metaboliese profiele van beide tradisionele en MG Kefir het gewys dat beide produkte asetaldehied, etanol, asetoon, diasetiel en asynsuur bevat. Die metaboliese profiele het verder gewys dat 'n onvoldoende diasetiel tot asetaldehied verhouding sowel as 'n tekort aan etielasetaat verantwoordelik was vir 'n geur defek in MG Kefir. Om hierdie defek te oorkom is sitraat en askorbaat (0.015% m.v-1) tydens Kefir fermentasie bygevoeg om diasetiel en etielasetaat produksie te verhoog. Hierdie byvoeging het 'n positiewe effek gehad op diasetiel produksie, maar nie op die produksie etielasetaat nie. 'n Verbetering in die algehele geur van Kefir is waargeneem aangesien die diasetiel tot asetaldehied verhoudings hoër (0.21 – 0.5) was in die monsters met bygevoegde sitraat en askorbaat as in die monsters daarsonder (0.12 – 0.17). Die produksie van etielasetaat in MG Kefir is verhoog deur die effekte van 'n verlengde inkubasie tydperk (24 h + 18 h by 22ºC), die byvoeging van etanol en asynsuur teen 0.79% (m.v-1) en die byvoeging van óf Lactococcus lactis ssp. diacetylactis biovar diacetylactis 318 óf Candida kefyr 1283 te kombineer. Die beste opbrengs is verkry in monsters wat C. kefyr 1283 en slegs etanol (9.22 mg.L-1) bevat het. Dit dui daarop dat etanol 'n belangrike faktor is vir etielasetaat produksie in Kefir beginstamme en wys moontlik op die afwesigheid van etielasetaat wat daarop dui dat die korrels nie 'n gis bevat wat bevoeg is om genoegsame hoeveelhede etielasetaat te produseer nie. Tydens hierdie ondersoek is die impak van etielasetaat op die organoleptiese kwaliteit van Kefir gedurende opberging by verkoelde- en kamertemperatuur ook bestudeer. Die resultate het gewys dat verkoelde Kefir tot 40 mg.L-1 etielasetaat bevat het sonder dat dit defektief was. Etielasetaat is dus beskou as 'n positiewe bydraer in terme van Kefir geur. Dit is van besondere belang aangesien etielasetaat 'n sterk geurkomponent teen konsentrasies laer as 5 mg.L-1 is. Verbeteringe in MG Kefir se geur was suksesvol en sal van waarde wees vir kommersiële Kefir produksie waar die hoofdoel die optimisering van Kefir geur is. Stabilisering van die korrels se mikrobiologiese konsortium is ook belangrik aangesien daar gevind is dat dit oor tyd verantwoordelik is vir stabiele en aanvaarbare Kefir. Die aanvaarbaarheid van tradisioneel, MG en ander Kefirs (Candi-Kefir en Lacto-Kefir), voorberei van mikrobiologies gestabiliseerde MG, is deur 85 verbruikers geëvalueer. Die resultate het aangedui dat pH (r = 0.978; p < 0.05) 'n belangrike faktor is in die bepaling van verbruikers se voorkeur van geur is, veral by vroulike verbruikers (r = 0.978; p < 0.05). Drie groepe, elk gekenmerk deur verskillende voorkeur en aanvaarbaarheid eienskappe, is verder geïdentifiseer. Groep I het oor die algemeen van geen van die produkte gehou nie en Groep III het die effense suur Kefirs soos Candi-Kefir (7.63) en Lacto-Kefir (7.09) verkies. Groep II het die Kefir met 'n matige suurheid en hoë etanolinhoud verkies. Tradisionele Kefir het die hoogste telling (7.50) en MG Kefir die laagste telling (4.78) behaal. Die sensoriese studie is van waarde aangesien dit gelei het tot die identifisering van die drywers van verbruikersvoorkeure deur die verskillende tipes verbruikers. Tydens hierdie projek is 'n naby-infrarooi reflektansie spektroskopiese metode ontwikkel vir die vinnige skatting van melk- en asynsuur, die organise sure wat verantwoordelik is vir die suurheid van Kefir, asook die pH en titreerbare suurheid (TS). Die resultate het getoon dat die kalibrasiemodelle vir melksuur (RPD = 2.57 – 3.16), pH (RPD = 2.90) en TS (RPD = 2.60) voldoende was vir siftingsdoeleindes (2 < RPD < 3). Dit dui daarop dat hierdie modelle sal aandui wanneer die konsentrasie van melksuur, pH of TS afwissel van die normale reeks. Hierdie studie het gewys dat die geur van MG Kefir, berei van verrykte korrels, suksesvol verbeter is en het ook gelei tot insigte in die voorkeur van aanvaarbaarheid van Kefir, 'n onbekende gefermenteerde suiwelproduk vir Suid-Afrikaanse verbruikers.

Face aux pathologies de malnutrition en croissance exponentielle en particulier dans les pays du Sud, le développement d’aliments fonctionnels à base de céréales fermentées représente une alternative aux produits laitiers déjà commercialisés. L’incorporation, la stabilité de composés bioactifs liposolubles tels que les caroténoïdes et les phytostérols, ainsi que leur devenir lors de la digestion gastro-duodénale doivent être étudiés afin de démontrer le potentiel nutritionnel de ce nouvel aliment. En effet, il est connu que les phytostérols diminuent la biodisponibilité des caroténoïdes. Le principal objectif de ces travaux a été de concevoir un aliment fonctionnel probiotique à base de maïs fermenté enrichi en caroténoïdes et phytostérols, et d’étudier la stabilité, la bioaccessibilité et l’absorption intestinale de ces composés liposolubles. La formulation et la standardisation du procédé, incluant la sélection de starters de fermentation afin d’obtenir un aliment probiotique, ont été mis au point. Au cours de la fabrication, la stabilité des composés liposolubles a montré un taux de rétention de 73 %. L’effet des phytostérols dispersibles sur la bioaccessibilité de la β-cryptoxanthine, du β carotène et du lycopène de l’aliment fonctionnel a été évalué à l’aide d’un modèle de digestion in vitro simulant les conditions gastriques et duodénales. L’absorption intestinale des composés a également été mesurée en couplant le modèle de digestion in vitro aux cellules de type Caco-2 (TC7). L’étude a démontré que grâce à leur véhicule, les phytostérols dispersibles apportent un potentiel hypocholesterolémiant au produit, sans affecter la bioaccessibilité des carotènes. In fine, cet aliment fonctionnel a été optimisé d’un point de vue organoleptique et nutritionnel par l’utilisation de protéines sériques
Because of the exponential growth of malnutrition pathologies particularly in South countries, the development of functional foods based on fermented cereals represents an alternative to dairy products already marketed. The incorporation, the stability of lipophilic bioactive compounds such as carotenoids and phytosterols, as well as their behavior during the gastro-duodenal digestion have to be studied, in order to demonstrate the nutritional potential of this new product. Indeed, it is known that phytosterols decrease the carotenoid bioavailability. The main objective of this work was to design a probiotic functional food based on maize and enriched with carotenoids and phytosterols, and study the stability, the bioaccessibility and the intestinal absorption of these fat-soluble compounds. The formulation and the standardization of the manufacturing process, including the selection of starters for fermentation in order to obtain a probiotic product, were developed. During the production, the stability of fat-soluble compounds showed a retention level of 73%. The effect on dispersible phytosterols on the β-cryptoxanthine, β-carotene and lycopene bioaccessibilities was evaluated thanks to an in vitro digestion model mimicking the gastric and duodenal conditions. The intestinal absorption of these compounds was also estimate by crossing the in vitro digestion model with a Caco-2 cells model (TC7). The study demonstrated that thanks to their vehicles, dispersible phytosterols provide a potential cholesterol-lowering effect to this food, without affecting the carotene bioaccessibility. In fine, this functional food was optimized in terms of sensory and nutritional levels, by using whey protein isolates

Duas formulações de queijo de leite cru e fermento endógeno foram produzidas e avaliadas após 30, 60 e 180 dias de armazenamento em câmaras de maturação de dois laticínios do Sudoeste do Paraná foram estudadas para avaliar as características, como uma das etapas de desenvolvimento de um queijo típico regional. A produção foi acompanhada desde a elaboração do fluxograma de processamento, coleta das amostras de queijo para realização de análises de proteína, lipídeos, umidade, cinzas, carboidratos, extrato seco total, gordura no extrato seco, calorias; atividade de água, textura instrumental (dureza, adesividade, elasticidade, coesividade, mastigabilidade e resiliência), cor (CIE Lab), análises microbiológicas (contagem de coliformes totais, contagem de coliformes termotolerantes e contagem de Staphylococcus coagulase positiva e pesquisa de Salmonella spp.); teste de aceitação relacionada às características sensoriais (cor, aparência, odor, textura, sabor) e intenção de compra. Essa pesquisa contribuiu com informações relevantes ao processo produtivo, tais como, a constatação da viabilidade do fermento lático liofilizado elaborado a partir das bactérias ácido láticas isoladas do leite da mesorregião do Sudoeste do Paraná e cujos resultados das análises dos queijos indicaram similaridade entre as formulações, no que se refere a caracterização física e físico-química, além de boa qualidade microbiológica, onde as discrepâncias entre as amostras dos laticínios não foram percebidas sensorialmente pelos provadores. Ajustes na padronização relacionados ao controle de qualidade tecnológica serão um fator de extrema importância para o sucesso das empresas e pequenos produtores envolvidos no projeto e que se dispõem a produzir o Santo Giorno, um queijo fino, com o grande diferencial de agregar características da região, com elevado padrão de qualidade higiênico-sanitária e com indicativos de grande aceitação pelo consumidor.
Two cheese formulations made from raw milk and endogenous yeasts with 30, 60 and 180 days of maturation in two dairy Paraná Southwest were studied to evaluate their quality through physical, physical-chemical, microbiological and sensorial characteristics, as one of the stages of development of a typical regional cheese. The production was accompanied from designing the flowchart processing, where the samples were collected to perform the analysis of proteins, lipids, moisture, ash, carbohydrates, total solids, fat in dry matter, calories; water activity, instrumental texture (hardness, adhesiveness, springiness, cohesiveness, resiliency and chewiness), instrumental color (CIE Lab); microbiological quality was assessed searching for total and thermotolerant coliforms, coagulase positive staphylococci and Salmonella spp.; the acceptance related to sensory characteristics of color, appearance, flavor, texture, taste and purchase intent was evaluated through the structured hedonic scale. This research contributed information relevant to the production process, such as the realization of the viability in freeze-drying lactic acid bacteria yeast isolated from milk in the Southwestern Mesoregion of Parana and the results of the analysis of the cheese showed similarity between formulations, regarding the physical, physical-chemical characterization, moreover good microbiological quality, where the differences between samples of dairy products were not perceived by sensory panelists. Adjustments in standardization related to technological quality control is an extremely important factor for the success of dairy companies and small producers involved in the project and that they have the option of producing the Santo Giorno, a fine cheese, with the great advantage of adding features of region, with high standard of sanitary conditions and with great consumer acceptance of indicative.

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CAPES
The aim was evaluate the effect of replacing corn by bakery waste (BW) in goats diet on performance, dry matter and nutrients intake, carcass characteristics, chemical composition and fatty acid profile of the Longissimus dorsi muscle and the food intake, apparent digestibility, nitrogen balance, ruminal parameters, production and composition of milk by goats. Experiment 1: Four levels of inclusion of BW replacing corn (0, 33, 66, 100%) in the diet of kids distributed in a completely randomized design were analyzed. The diets were composed of concentrate and Cynodon spp. hay, with forage: concentrate ratio of 60:40. The intakes of dry matter, organic matter, crude protein, ether extract and carbohydrates decreased linearly, while the non-fiber carbohydrates intake, animal performance, carcass characteristics and chemical composition were not affected by replacement of corn by BW. The elaidic acid content (C18: 1 trans-9) increased in the fatty acid profile of the Longissimus dorsi muscle with the inclusion of RP, which can be used as a substitute for corn meal in goats feed composition. It was concluded that bakery waste can substitute the corn up to 100% in the concentrate mixture without affecting intake, performance and carcass and meat traits of kids. Experiment 2: Bakery waste was added replacing 0; 25; 50; 75 and 100% of the concentrate in dry matter basis. Five Saanen lactating goats, non-pregnant with an average weight of 55.7 kg were arranged in a 5 x 5 Latin Square design. Experimental diets were composed by Cynodon hay and concentrate, in a roughage:concentrate ration of 40:60. The inclusion of BW in diets decreases the intake of ether extract, acid detergent fiber and the coefficients of apparent digestibility of crude protein and ether extract. Different levels of substitution did not affect ruminal pH, but for the concentration of ruminal ammonia was found linear reduction with the level of replacement. The inclusion of BW in diets increased the efficiency of use of N. The bakery waste can replace conventional concentrate in diets for goats.
Objetivou-se avaliar o efeito da substitui??o do milho pelo res?duo de panifica??o (RP) na dieta de cabritos sobre o desempenho, consumo de mat?ria seca e dos nutrientes, caracter?sticas de carca?a, composi??o qu?mica e perfil dos ?cidos graxos do m?sculo Longissimus dorsi e em cabras em lacta??o o consumo alimentar, a digestibilidade aparente, o balan?o de compostos nitrogenados, os par?metros ruminais, a produ??o e a composi??o do leite. Experimento 1: Foram analisados quatro n?veis de inclus?o do RP em substitui??o ao milho (0, 33, 66, 100%) na dieta de cabritos distribu?dos em delineamento inteiramente casualizado. As dietas foram compostas de concentrado e feno de Cynodon spp, com rela??o volumoso:concentrado de 60:40. O consumo de mat?ria seca (MS), mat?ria org?nica (MO), prote?na bruta (PB), extrato et?reo (EE) e carboidratos totais (CT) foi linear decrescente, enquanto que o consumo de carboidratos n?o fibrosos (CNF), o desempenho dos animais, as caracter?sticas de carca?a e a composi??o centesimal n?o foram influenciados pela inclus?o do RP em substitui??o ao milho. O teor de ?cido ela?dico (C 18:1 trans-9) aumentou no perfil de ?cidos graxos do m?sculo Longissimus dorsi com a inclus?o do RP. Conclui-se que o RP pode substituir o milho em at? 100% na mistura do concentrado. Experimento 2: Foram avaliados cinco n?veis de substitui??o do concentrado pelo RP (0, 25, 50, 75, 100%), utilizando-se cinco cabras em lacta??o da ra?a Saanen, com peso m?dio de 55,7 kg, distribu?dos em delineamento em Quadrado Latino 5 X 5. As dietas experimentais foram compostas de concentrado e feno de Cynodon spp, numa rela??o volumoso:concentrado de 40:60 na mat?ria seca. Houve redu??o linear do consumo de EE, do consumo de FDA, da digestibilidade da PB e da digestibilidade do EE. A substitui??o crescente do concentrado pelo RP na dieta n?o influenciou o pH ruminal, por?m reduziu a concentra??o de N-NH3 no l?quido ruminal. A inclus?o do RP em n?veis crescentes nas dietas favoreceu a efici?ncia de utiliza??o de nitrog?nio. O RP pode substituir totalmente o concentrado na dieta de cabras em lacta??o.

La demanda creixent de productes mínimament processats i llestos per al consum planteja un important repte per a la seguretat alimentaria i ha conduit al desenvolupament de tractaments suaus que permetin inhibir el creixement microbià conservant la qualitat dels aliments. Els treballs recollits a la present tesi van plantejar diverses estratègies consistents en la combinació d'obstacles al creixement microbià per a millorar la seguretat de productes carnis llestos per al consum. Amb l'objectiu de millorar la seguretat dels embotits poc àcids, es va valorar l'aplicació del tractament per alta pressió hidrostàtica (APH) i l'addició de cultius iniciadors en embotits poc àcids. Per altra banda, per a reduir el risc de L. monocytogenes durant la conservació del pernil cuit llescat, es va avaluar l'efecte combinat de l'addició d'antimicrobians naturals (lactatat-diacetat i enterocines), afegits directament o a través de l'envasament antimicrobià, i el tractament per alta pressió hidrostàtica.
The growing demand of minimally processed ready to eat food products raises a challenge for the food safety and has derived in the development of mild treatments effective to inhibit microbial growth without damaging food quality. The articles collected in this work studied the application of some strategies consisting of a combination of hurdles to inhibit microbial growth in order to improve the safety of ready to eat meat products. On one hand, the combined effect of high pressure processing and addition of starter cultures to improve the safety of low acid fermented sausages (fuet and chorizo) was assessed. On the other hand, the combination of high pressure processing and addition of natural antimicrobials (lactate-diacetate and enterocins), added directly or included in antimicrobial packaging, in order to reduce the risk of L. monocytogenes during storage of sliced cooked ham was evaluated.

Fermentation is one of the oldest techniques for the production of foods. Lactic acid bacteria (LAB) have been progressively used in food industry since, besides improving nutritional and technological features, they also contribute to the safety of food products. Consequently, this work aimed to isolate lactic acid bacteria from several food products and to evaluate their potential antimicrobial activity and probiotic characteristics. From 202 LAB isolated from 20 fermented food products, only three isolates were selected and identified as Enterococcus faecium RS7 (fermented shrimp), Enterococcus faecium P12 (pork sausage) and Leuconostoc lactis RK18 (khmer fermented rice fresh noodles) based on their antimicrobial activity against Enterococcus faecalis and Listeria monocytogenes strains. These selected isolates were further tested for probiotic characteristics. Despite the beneficial characteristics apparently presented by the three isolates due to their antimicrobial activity, only Ln. lactis RK18 met the safety requirement and therefore it was the only isolate selected for further tests. Leuconostoc lactis RK18 did not present any of the virulence factors nor virulence genes tested (with exception of aggregation substance protein gene asa1), and it was also susceptible to all antibiotics recommended by European Food Safety Authority. Regarding beneficial characteristics, it was found that anti-listerial activity of Ln. lactis RK18 was due to the production of a Class IIa bacteriocin (<6.5 kDa in size), which remained stable at average temperatures (30 ºC to 80 ºC) and at pH values ranging from 4 to 6, and although susceptible to some detergents, it showed greatly resistance to several enzymes. Despite being very sensitive to acidic environments, when incorporated into a complex food matrix such as alheira, Ln. lactis RK18 was able to survive through simulated gastrointestinal tract (GIT) conditions and also to adhere (but not invade) to human colon adenocarcinoma cell lines Caco-2 in vitro. Even though exposure to GIT conditions had influenced the adhesion ability of Ln. lactis RK18 cells, this potential probiotic and merely 10% of its treated cell-free supernatant, were able to prevent the ability of L. monocytogenes CEP 104794 to adhere and invade Caco-2 cells. Overall, Ln. lactis RK18 appeared to be a safe strain, with no risk to human health, which harbored important features to be successfully considered as a potential biopreservative and probiotic culture. Nevertheless, further experiments should be performed for the validation of its application in the food industry.
A fermentação é uma das mais antigas técnicas na produção de alimentos. As bactérias do ácido lático (BAL) têm sido muito usadas na indústria alimentar, uma vez que melhoram as características nutricionais e tecnológicas, e contribuem para a segurança dos alimentos. Assim, o objetivo deste trabalho foi o isolamento de BAL de vários alimentos e de avaliar a sua potencial atividade antimicrobiana e características probióticas. De 202 BAL isoladas de 20 alimentos fermentados, apenas três isolados foram selecionados e identificados como Enterococcus faecium RS7 (camarão fermentado), Enterococcus faecium P12 (linguiça de porco) e Leuconostoc lactis RK18 (khmer “noodles” de arroz fermentado fresco), seleção essa baseada na sua atividade antimicrobiana contra estirpes de Enterococcus faecalis e Listeria monocytogenes. Os isolados selecionados foram testados quanto a características probióticas. Apesar da atividade antimicrobiana apresentada pelos três isolados, apenas Ln. lactis RK18 preencheu o requisito de segurança e, portanto, foi o único isolado selecionado para outros testes. Leuconostoc lactis RK18 não apresentou nenhum dos fatores ou genes de virulência testados (exceto o gene da proteína de substância de agregação asa1), e também foi suscetível a todos os antibióticos recomendados pela Autoridade Europeia de Segurança Alimentar. Relativamente às características benéficas, a atividade anti-listeria de Ln. lactis RK18 foi devida à produção de uma bacteriocina Classe IIa (tamanho <6,5 kDa), a qual permaneceu estável a temperaturas moderadas (30 ºC a 80 ºC) e em valores de pH entre 4 a 6 e, embora sensível a alguns detergentes, apresentou grande resistência a várias enzimas. Apesar de muito sensível a ambientes ácidos, quando incorporado numa matriz alimentar complexa como a alheira, Ln. lactis RK18 sobreviveu às condições simuladas do trato gastrointestinal (TGI) e aderiu (mas não invadiu) às linhas celulares de adenocarcinoma do cólon humano Caco-2 in vitro. Embora a exposição às condições do TGI tenha influenciado a capacidade de adesão de Ln. lactis RK18, este potencial probiótico e apenas 10% do seu sobrenadante, preveniram a adesão e invasão de células intestinais por L. monocytogenes CEP 104794. Em conclusão, Ln. lactis RK18 parece ser uma estirpe sem risco para a saúde humana, com características importantes que a tornam uma potencial cultura bioconservante e probiótica. No entanto, mais experiências devem ser realizadas para a validação da sua aplicação na indústria alimentar.

The incidences of diarrhoeal episodes in infants and children have mostly been associated with the consumption of contaminated weaning foods. This is especially true in developing countries where factors such as the lack of sanitation systems and electricity have been found to contribute to an increase in the incidence of microbiologically contaminated weaning foods. The process of fermentation has been found to reduce the amount of microbiological contamination in such foods as a result of the production of antimicrobial compounds such as organic acids, peroxides, carbon dioxide and bacteriocins. In this study, microbiological surveys were conducted on sorghum powder samples and their corresponding fermented and cooked fermented porridge samples collected from an informal settlement of the Gauteng Province of South Africa. The process of fermentation was found to result in significant decreases (P>0.05) in Gram-negative counts and spore counts, while aerobic plate counts decreased slightly. Lactic acid bacteria counts, however, increased significantly (P>0.05). The cooking process was found to result in further significant decreases (P>0.05) in all counts. Sorghum powder samples and fermented porridge samples were found to be contaminated with potential foodborne pathogens, including Bacillus cereus, Clostridium perfringens and Escherichia coli, however, none of the pathogens tested for were detected in any of the cooked fermented porridge samples. SDS-PAGE and phenotypic analysis of 180 lactic acid bacteria isolated from sorghum powder samples and their corresponding fermented and cooked fermented porridge samples showed that a majority of the isolates were lactobacilli and leuconostocs, however, some isolates were identified as pediococci and lactococci. These results demonstrated the heterogeneity of the lactic acid bacteria isolates that were associated with fermentation processes in this study. Of the lactic acid bacteria identified, Lactobacillus plantarum and Leuconostoc mesenteroides strains were found to have the highest distribution frequencies, being distributed in 87% and 73% of the households, respectively. Analysis of Lactobacillus plantarum (58) and Leuconostoc mesenteroides (46) strains isolated from sorghum powder samples and corresponding fermented and cooked fermented porridge samples by AFLP fingerprinting showed that they originated from a common source, which was sorghum powder. There was, however, evidence of strains that may have been introduced at household level. Antimicrobial activity of selected lactic acid bacteria was found to be mainly due to a decrease in pH in fermented and cooked fermented porridge samples. None of the lactic acid bacteria tested seemed to produce bacteriocins.

碩士
南台科技大學
化學工程系
91
Recent studies demonstrated that many natural foods have ability to restrain tumor cells, because the compounds of these foods contained can inhibit the growth of human cancer cells or induce the death of cancer cells. The fruiting body of Antrodia camphorata are said to be effective in liver therapy in folklore. The aim of this study is to cultivate Antrodia camphorata mycelium with various natural foods by the solid-state and submerged fermentation, and then the extract of fermented mycelia, fermented broth and the foods are tested by MTT assay to investigate the effectiveness on cancer cells inhibition. The results showed that the alcohol extract of fermented food is more effective in suppressing the growth of cancer cells than the extract of foods. The extract of different fermented foods posses different degree of inhibition to human cancer cells proliferation. Low concentration（0.4%）extracts of A. camphorata by solid-state culturing can effectively inhibit the proliferation of different human cancer cells. In contrast, much higher extract concentrations of A.camphorata by submerged fermentation are needed to reach the same inhibition effect. From the microscopic observation, the extracts of A. camphorata by solid-state culture can induce tumor cell death by the mechanism seeming to similar to apoptosis. The results indicated that some natural foods metabolized by A. camphorata can produce cytotoxic compounds that are detrimental to tumor cells.

碩士
國立臺灣海洋大學
食品科學系
92
Abstract Tilapia were fermented with 4% Sucrose, 1% Glucose, 2% NaCl, 50% Distilled water, and lactic acid bacteria (LAB) strains of Pediococcus (Ped.) pentosaceus MFL, Ped. pentosaceus MFS, Lactobacillus (Lb.) plantarum BCRC12250, or their combination, and non-starter. LAB were as starters in tilapia meat paste. After 12 hr fermentation, the pH value rapidly declined to under 4.6 and cut down fermented time to 12 hr. The LAB growth rapidly, became predominant during fermented tilapia meat paste period and the growth of other microorganisms was substantially inhibited. VBN of sample fermented with LAB were slow increased after 24 hr fermentation. The gel strength of the 24 hr fermented tilapia meat paste as LAB was used the starters was 621.08-742.74 cm x g and the gel strength obtained from the 24 hr fermented tilapia meat paste without LAB was 429.48 cm x g. These results may suggest that fermented tilapia meat paste with the addition of LAB can enhance the gel strength ability. Between fermented tilapia meat paste with/without the addition of LAB, its Hunter a and b value had no significant difference. But the whiteness of samples with the LAB fermented tilapia meat paste were significantly higher than without starter (whiteness: from 51.71 increased to 58.34 and 61.98). The results of SDS-PAGE of LAB fermented tilapia meat paste suggested that most of water- and salt-soluble muscle protein were rapidly degraded after 12 hr fermentation and increased in amounts of free amino acid. In the evaluation on the antioxidation effect of the cold water extracts of fermented tilapia food (50 mg/mL), the ability for scavenging DPPH free radical was examined to perform 40-94%. Scavenging DPPH free radical of the cold water extracts of fermented tilapia food by Ped. pentosaceus MFL and MFS starters demonstrated 94% antioxidative activity. The results indicated that the cold water extracts of five tilapia fermented food performed 68-98% chelating effect by assaying the ability of chelating on Fe (II) ion. The antioxidative effect was measured via the inhibition of hemoglobin-induced linoleic acid oxidation. The cold water extracts of fermented tilapia food by using Ped. pentosaceus MFL and MFS starters significantly obtained 81% antioxidative activity. Reducing power of the cold water extracts of fermented tilapia food with LAB were performed well to A700nm = 0.85-0.92. Four groups of the cold water extracts of fermented have good performance on inhibiting S. typhimurium TA100 mutation induced by 4NQO or B[a]P, the antimutagenicity observed were 12-26% and 28-55%, respectively. The antimutagenicity ability of exhibited by B[a]P was better than 4NQO. The antimutagenicity ability of the cold water extracts of tilapia and without starter fermented were not so pronounced, especially for without starter group the mutation inhibition capability was below 15%. The growth-promotion ability of four groups of the cold water extracts of fermented tilapia food with LAB to immuno-cell line, HB4C5 and THP-1, showed significant difference to the cold water extracts of tilapia and without starters. The ACE inhibiting activity ability of four groups from the cold water extracts of fermented with LAB was the better to perform (IC50 = 0.06-29.47 mg/mL) than without starters (IC50 = 16.70 mg/mL), especially for the cold water extracts by using Ped. pentosaceus MFL and MFS can reach IC50 = 0.06 mg/mL

Micro-organisms possess endogenous enzymes, however, the stability of these enzymes during storage in soymilk has not been studied. β-glucosidase is an important enzyme that could be used in the bioconversion of the predominant soy isoflavone glycosides to their bioactive aglycone forms. Fifteen probiotic micro-organisms that included Bifidobacterium sp., Lactobacillus acidophilus and Lactobacillus casei were screened for β-glucosidase activity using ρ- nitrophenyl-β-D-glucopyranoside as a substrate. Six strains were selected on the basis of β- glucosidase activity produced during fermentation of soymilk. The stability of the enzyme activity was assessed during incubation for up to 48 h and during storage for 8 weeks at frozen(-80°C), refrigerated (4°C), room (25°C) and incubation (37°C) temperatures. L. casei strains showed the highest β-glucosidase activity after 24 h of incubation followed by L. acidophilus strains, while Bifidobacterium strains showed least activity. However, β-glucosidase from Bifidobacterium animalis BB12 showed the best stability during the 48 h fermentation. Lower storage temperatures (-80°C and 4°C) showed significantly higher (P<0.05) β-glucosidase activity and better stability than that at higher temperatures (25°C and 37°C). The stability of β-glucosidase from these microorganisms should be considered for enzymic biotransformation during storage, of isoflavone β-glycosides to bioactive isoflavone aglycone forms with potential health benefits. Three strains of L. acidophilus, two strains of L. casei and one strain of Bifidobacterium were screened for β-glucosidase activity using ρ-nitrophenyl-β-D-glucopyranoside as a substrate and their potential for the breakdown of isoflavone glycosides to the biologically active aglycones in soymilk. Isoflavones quantification with HPLC and β-glucosidase activity was performed after 0, 12, 24, 36, and 48 h of incubation in soymilk at 37°C. All 6 microorganisms produced β-glucosidase, which hydrolysed the predominant isoflavone β-glycosides. There was a significant increase in the concentration of isoflavone aglycones and a subsequent decrease (P < 0.05) in the concentration of isoflavone glycosides in fermented soymilk. Based on the concentration of isoflavones during peak β-glucosidase activity, the hydrolytic potential was evaluated. L. acidophilus 4461 had the highest aglycone concentration of 76.9% after 24 h of incubation, up from 8% in unfermented soymilk (at 0 h). It also had the best isoflavone hydrolytic index of 2.01, signifying its relative importance in altering the biological activity of soymilk. Soymilk fermented containing soy isoflavones with B. animalis Bb12 was stored at various temperatures (-80°C, 4°C, 25°C and 37°C) for 8 weeks and the concentration of isoflavones determined weekly using RP-HPLC. The first order kinetic model was used to assess the degradation of each isoflavone isomer at each storage temperature. During storage at various temperatures, concentrations of individual isoflavone isomers appeared to be significantly stable (P<0.01). Interestingly, the aglycones showed much smaller degradation constants as compared to the glycosides at all the storage temperatures. Genistein and daidzein were much more stable than glycitein and had almost similar degradation pattern, despite differences in their concentrations in the fermented soymilk. It was, however, observed that 4°C was the most suitable storage temperature for the product as there was a minimal degradation of bioactive isoflavone aglycones. Three selected L. acidophilus strains were used in the fermentation of soymilk and then stored separately at various temperatures (-80°C, 4°C, 25°C and 37°C) for 8 weeks and the concentration of isoflavones determined weekly using RP-HPLC with diode array uv visible detector. The decreasing concentration of isoflavones in soymilk during storage due to degradation was found to fit first order kinetic model. Isoflavone aglycones as well as isoflavone glycosides largely appeared to be stable during storage (P<0.01). Interestingly, the aglycone forms showed much smaller degradation as compared to glycoside forms at all the storage temperatures. Of the isoflavone aglycones, daidzein was most stable followed by genistein, while glycitein was least stable. Isoflavone aglycones such as glycitein, daidzein and genistein showed smaller degradation constants in fermented soymilk at lower storage temperatures (-80°C and 4°C) and higher degradation constants at higher storage temperatures (25°C and 37°C) with each strain. In contrast, glycosides glycitin and daidzin showed higher degradation at lower storage temperatures (-80°C and 4°C) and lower degradation at higher storage temperatures (25°C and 37°C). Storage temperature was therefore found to be very important in regulating the rate of degradation of soy isoflavones in fermented soymilk. The degradation of each isoflavone compound in soymilk fermented with 2 Lactobacillus casei strains and stored at various storage temperatures (-80°C, 4°C, 25°C and 37°C) was evaluated and again found to fit the first order kinetic model. All isoflavone compounds in the soymilk appeared to be generally stable during storage (P< 0.01) at all storage temperatures. Aglycone forms however, had smaller degradation constants compared to glycosides at all storage temperature in the presence of each of the micro-organisms. Specifically, aglycones showed a unique trend of smaller degradation at lower storage temperatures (-80ºC and 4ºC) than at higher temperatures (25ºC and 37ºC). Glycoside genistin was least stable at all storage temperatures compared to other isoflavones, while aglycone daidzein was the most stable. L. casei 2607 in fermented soymilk stored at 4ºC after 8 weeks gave the least degradation for daidzein of a mere 3.78% loss from 9.53 to 9.17 ng/µL. L. casei 2607 showed greater hydrolytic potential than L. casei ASCC 290 as denoted by higher degradation of isoflavone glycosides in fermented soymilk at lower storage temperatures. The optimum storage temperature offering least degradation of bioactive isoflavone aglycones in fermented soymilk was found to be 4ºC. Liquid chromatography coupled with positive electro spray ionisation tandem mass spectrometry (MS/MS) and diode array detection was used for the quantitation and characterisation of isoflavones in fermented and unfermented soymilk made from soy protein isolate SUPRO 590. Bifidobacterium animalis ssp. lactis Bb12 was used for the fermentation of soymilk. The isoflavones were found to produce characteristic radical ions as well as molecules of H20, CO2, a sugar unit, and an alcohol through collision-induced fragmentation. Product ion fragments revealed unique fragmentation pathways for each isoflavone compound. Characteristic fragmentation of different isoflavones were unequivocally identified and differentiated. The occurrence of aldehydes such as pentanal, ethanal and methanal was shown to be specifically linked with isoflavone aglycones, daidzein, genistein and glycitein, respectively. Main glycosides such as genistin, daidzin and glycitin as well as the acetyl-, and malonyl forms also showed respective aglycone ions in their spectra fragmentation. Thus positive ion fragmentation was important in the unequivocal confirmation of isoflavones and for revealing the occurrence of other related compounds such as aldehydes in the soymilk. Comparison of endogenous β-glucosidases and β-galactosidases in selected probiotic bacteria as hydrolysing enzymes in the breakdown of the predominant isoflavone glycosides in soymilk into bioactive isoflavone aglycones is critical for an optimised processing of a probiotic functional food. β-glucosidase activity and β-galactosidase activity of probiotic organisms including L. acidophilus ATCC 4461, L. casei 2607 and B. animalis ssp. lactis Bb12 in soymilk was evaluated and correlated with the increase in concentration of isoflavone aglycones during fermentation. The concentrations of isoflavone compounds in soymilk were monitored using a Varian model HPLC with an amperometric electrochemical detector. In all microorganisms, β-glucosidase activity was found to be greater than that of β-galactosidase. The aglycone concentration in the soymilk with L. acidophilus 4461, L. casei 2607 and B. animalis ssp. lactis Bb12, increased by 5.37, 5.52 and 6.10 fold, respectively after 15 h of fermentation at 37ºC. The maximum hydrolytic potential was also observed at 15 h of fermentation for the three micro-organisms coinciding with peak activities of the two enzymes. β-glucosidase activity was found to be more than 15 times higher than that of β-galactosidase activity in the soymilk for each microorganism during fermentation. It appears β-glucosidase played a greater role in isoflavone hydrolysis. It is important to determine critical parameters such as which hydrolysing enzyme have a greater impact in the development of a probiotic functional food beverage. In this case it is essential to enhance β-glucosidase activity for its greater role in improving the biological activity of soymilk during processing. Having established that endogenous β-glucosidase plays a greater role in isoflavone biotransformation, it was essential to compare endogenous and exogenous β-glucosidases for their role in isoflavone biotransformation. β-glucosidase activity of probiotic organisms including Bifidobacterium animalis ssp. lactis Bb12, Lactobacillus acidophilus ATCC 4461 and Lactobacillus casei 2607 in soymilk was evaluated and found to relate to the increase in concentration of isoflavone aglycones during fermentation. The concentrations of isoflavone compounds in soymilk were monitored using a Varian model HPLC with an Amperometric electrochemical detector. The aglycone composition, also known as aglycone equivalent ratio, has been considered to be important for delivery of health benefits of isoflavones, was also monitored during fermentation of soymilk. Comparison of the hydrolytic effectiveness of both exogenous and endogenous enzyme during 4 h incubation in soymilk was conducted using the Otieno-Shah (O-S) index. Results showed that exogenous enzyme exhibited faster rate of isoflavone glycoside hydrolysis than that by endogenous enzyme. Highest O-S indices were obtained after 4, 3 and 2 h of incubation with enzyme solution having β-glucosidase activity of 0.288 UmL-1, 0.359 UmL-1, and 0.575 UmL-1 resulting into aglycone concentration increments of 5.87, 6.07 and 5.94 fold, respectively. Conversely, aglycone concentration in the soymilk with B. animalis ssp. lactis Bb12, L. casei 2607 and L. acidophilus 4461 increased by 3.43, 2.72 and 3.03 fold, respectively after 4 h of fermentation at 37ºC. Also, the O-S index of endogenous enzyme was much lower than that of the exogenous enzyme over the same 4 h incubation period. Optimum aglycone equivalent ratios coincided with highest O-S indices and highest aglycone concentrations in soymilk hydrolysed with exogenous enzyme. The same correlation of O-S indices and highest aglycone concentrations occurred for endogenous enzyme during the 24 h of fermentation. Therefore, obtaining highest aglycone concentration as well as optimum aglycone equivalent ratio could provide a critical beginning point in clinical trials for maximum realisation of the unique health benefits of soy isoflavones. Screening for β-glucosidase activities of probiotic bacteria in soymilk as well as comparing their hydrolytic potentials with that exogenous β-glucosidase could find wide applications in the development of different aglycone rich functional soy beverages.

Fermentation is a process by which primary food products are modified biochemically by the action of microorganisms and/or their enzymes. Several societies have, over the years, intentionally carried it out to enhance the taste, aroma, shelf-life, texture, nutritional value and other properties of food. It is used in many parts (lithe world. However, there are regional differences in use and these depend on the availability of raw materials, consumption habits. and other socio-cultural factors. This study was aimed at (comparatively) assessing, local commercial and homemade amahewu with respect to nutritional value, hygiene and other health benefits to the commirn ity. Methods employed were Thin Layer Chromatography (TLC) (mycotoxins), High Perliffmance Liquid Chromatography (HPLC) (mycotoxins, sugars and amino acids), Dumas (proteins), SOxhlet (lipids) and intubation technique (metabolisable energy) to analyse maize meal and amahewu samples from various regions. The regions sampled included mal3heleni (South Coast) and kwaNgcolosi (North Coast) villages. Commercial amahewu was analysed with kind permission from Clover SA. Species from the following genera were isolated and identified from amahewu samples: Lactobacillus, Saccharonivccs, Lcuconostoc, Lactococcus, Panioca, Entcrobacter and kleb•iella. Saccharotnyces was detected in commercial samples only. Gram-negative strains were identified in most of manheleni village samples. No traceable amounts of aflatoxin BI (AFB1), fumonisin B 1 (FBI) and zearalenone (ZEA) were found in Clover SA samples. AFB I was detected in 40% of both maize meal and amahewu samples from maBheleni (range 0.55 — 0.84ng/g and 8.3x10 5 — 9.1x10-5ng/g respectively). From the same village, 100% of the maize meal and 80% of the amahewu samples were contaminated with FBI (range 4.1 47.2ng/g and 1.4 ---- 6.9ng/g respectively). ZEA was detected in all maize meal samples (range 0.9 — 4.3ng/g). None of the amahewu samples contained detectable levels of ZEA. All maize meal and amahewu samples from kwaNgcolosi were contaminated with AF13 1 (range 8.3 — 30.I ng/g and 0.04 - 0.102ng/g respectively). FB I was detected in 75% of both maize meal and amahewu samples from the same village (range 0.5 — 4.1ng/g and 0.04 0.56ng/g respectively). ZEA was also found in all maize meal samples and 75% of amahewu samples (range 3.7 — 16.4ng/g and 0.03 -- 0.06ng/g respectively). MaBheleni, Clover SA and kwaNgcolosi maize meal and amahewu samples contained vitamins B1, 13 2 and B6 with a range of 0.31+0.21 - 4.48±0.81 B 1 ; 0.15±0.14 - 1.67±0.33 B2 and 0.05±0.07 - 0.77±1.45 lig/g B6. Fat levels ranged from 0.28±0.40 to 4.54±0.05 percentage by weight. The levels of proteins varied from 4.02±0.02 to 8.40±0.04 percentage by weight. Starch concentrations ranged from 31.51.5.28 to 75.911.92g/100g. Maize meal samples contained glucose and maltose, while glucose, fructose, sucrose, maltose, M-triose, DP 4 and 5 and DP >15 were detected in amahewu. Apparent and true metabolisable energy for homemade and commercial Freeze-dried amahewu was 13.194 and 13.696MJ/kg (AME N ); and 13.605 and 14.106M.Ekv ( 1 MEN ), respectively. This study has shown that lactic acid maize fermentation reduce' the levels of AF13 1 , FB I and ZEA toxins in maize meal, inhibits the growth of most Gram-negative bacteria, and in some instances, fermentation did improve the nutritional value. Metabolisable energy analysis represents an important tool to assess whether or not compounds ingested are converted to sources of energy in the body and utilised. Amahewu fermentation yielded beneficial products (probiotics: reduced mycotoxins levels and reduced starch). In conclusion, natural lactic acid maize fermentation to produce amahewu will do more good than harm to the consumer, therefore, people need to be advised on how to safely store their maize and also to be encouraged to consume their stored maize in fermented form.
Thesis (M.Med.Sc.)-University of Natal, Durban, 2003.

Submitted in fulfilment of the academic requirements for the Degree in Master's in Food Science and Technology, Durban University of Technology, 2017.
Pigeon pea (Cajanus Cajan) is consumed in many parts of Africa as a source of protein and carbohydrate. It is underutilised and mainly grown for subsistence. Researching on pigeon pea may enhance value addition and increase its utilization. In this study, a non-dairy probiotic yoghurt was prepared from pigeon pea milk. Yoghurt samples were prepared, using 100% pigeon pea milk, pigeon pea/soy milk in the ratio 50:50 and 100% soy bean milk. The yoghurts were inoculated with yoghurt starter cultures and divided into two equal parts. One part inoculated with Propionibacterium freudenreichii was referred to as probiotic yoghurt, while the other part served as the control. The nutritional, sensory and some functional properties of the yoghurt were determined. The microbiological quality of yoghurt samples stored at 4, 10 and 21°C, respectively, for 4 weeks, were monitored and analysed for aerobic spores’ formers, E. coli, total plate counts, mould and Propionibacterium freudenreichii weekly. The protein contents of the yoghurt samples varied from 4.54-5.85% for 100% soymilk and 100% pigeon pea yoghurt respectively. The probiotic yoghurt showed slightly lower protein content than pigeon pea yoghurt alone. All the yoghurt samples had considerably high total solids (16.04-17.41%) and were fairly good sources of amino acids. Essential amino acids in the yoghurt samples were comparable to the FAO/WHO (2007) recommended amino acid requirement for adults. Anti-nutritional factors of yoghurt samples were significantly lower (P≤0.05) than their milk counterparts, which may be attributed to the fermentation process. Probiotic yoghurt samples showed higher firmness than non-probiotic samples. Total plate counts (log 7.01- 7.46 CFU/ml) samples stored for 2 weeks at 4° C were similar. Predominant organisms were LAB and Propionibacterium freudenreichii. Storage temperature of yoghurt samples had an influence on the total plate count and LAB. Total plate count and LAB significant increased approximately by log 2 CFU/ml for the first two weeks of storage. However, moulds and E. coli were not detected in all samples. Beyond 2 weeks of storage, there was significant decline in total plate counts and LAB, while mould grew and increased. Aerobic spore formers and moulds were observed in the control yoghurt. However, E. coli was not found in all yoghurt samples throughout storage period. The pH of the milk in which yoghurt mixtures were formulated, ranged from pH 7 to 6.8 for pigeon pea and soymilk declined significantly as a result of acidification. Decline in pH at 4, 10 and 21°C was significant (p≤0.05) with the rate higher at 21, 10 than 4° C. Decline in pH resulted in increased TTA values over storage temperatures and periods. Samples stored at 21°C and 10°C had significantly higher TTA values than samples stored at 4° C. The colour values evaluated were recorded as L*, b*, a* and ∆E* during 4 weeks storage at 4, 10 and 21° C. Significantly high values (p≤0.05) were recorded for L* yoghurt samples with soymilk. The colour scale defines positive (red) and negative (green) for a* and b* positive (yellow) and negative (blue). All a* values both positive and negative were less than 3. There was no negative value recorded for b*. Colour difference ∆E* values trends increased as storage time and temperature increased. There were significant (p≤0.05) differences between samples stored at same and different storage temperatures and periods. Water holding capacity was significantly different (p≤0.05) in all the yoghurt samples stored at 4, 10 and 21°C for 4 weeks. Formulation with 100% soymilk recorded higher values. Soy yoghurt and probiotic yoghurts (100 %) showed higher water holding capacity compared to pigeon pea yoghurt and pigeon pea/soymilk yoghurt. The addition of Propionibacterium freudenreichii did not significantly affect sensory properties of the yoghurts. Acceptable yoghurt was produced from pigeon pea with comparable quality to soy which serves as control. Proximate composition was comparable to previous reports. Microbial quality and profile of all the yoghurt samples were similar. The absence of pathogenic bacteria in all the yoghurt samples confirm their safety. Soy yoghurt was most acceptable amongst the yoghurt samples but all the samples had comparable ratings, and these ratings are within commercially acceptable range (4 to 9) for yoghurt. Storage at 4oC should be the most acceptable, as storage at 21oC encourage proliferation of contaminant
M

PhD (Food Science)
Department of Food Science and Technology
Finger millet (FM) [Eleusine coracana] is an underutilised cereal grain used as a food source in South Africa. Increased research interest in FM has span over the years owing to its unique nutritional and bioactive composition. Following the recent interest in natural curative substances over their synthetic counterparts in the treatment of food dependent diseases, FM has shown potential nutraceutical effects. Some important health effects like antidiabetic, antioxidative, anti-inflammatory and antimicrobial properties have been reported in recent trials with FM. In view of the increasing utilisation and application of FM in the region of Thulamela Municipality, Vhembe District of South Africa, two common indigenous FM varieties (brown and dark brown) were obtained and analysed for their physicochemical properties, levels of minerals, phytic acid, phenolic compounds and antioxidant activities. For this process, malted non-alcoholic beverages were produced and analysed for their physicochemical properties, levels of phenolic compounds, and total phenolics and antioxidant activities. FM grains were soaked, germinated and kilned at an interval of 24 h for 96 h, using sorghum as an external reference. Mineral composition of the FM and sorghum samples were analysed using an inductively coupled plasma atomic emission spectroscopy (ICP-AES) and mass spectroscopy (ICP-MS), and atomic absorption spectrometer (AAS). Identification and quantification of phenolic compounds were performed using ultra-performance liquid chromatography mass spectrometer (UPLC-MS). All experiments were performed in triplicate except for the UPLC-MS analysis of the malted non-alcoholic beverages that was done in duplicate. Data were analysed by one way analysis of variance, and the mean values were separated by Duncan’s multiple comparison test using SPSS version 24.0. Data showed that the FM varieties were rich in macro- and micro- or trace elements. The macro-elements calcium, magnesium, potassium, phosphorus and sulphur were found in high amounts ranging from 1597.37 mg/ kg – 6775.03 mg/ kg; iron, zinc, strontium and silicon were found in significant amounts in the range 21.47 mg/ kg – 55.67 iii mg/ kg, copper and boron were found in low amounts (2.2 mg/ kg – 7.7 mg/ kg), along with selenium and cobalt (0.02 mg/ kg – 0.05 mg/ kg). Heavy metals, barium and aluminium were found in the FM varieties. Varietal difference was found to play an important role in the mineral content of the grains during malting. Malting for 24 h reduced mineral content except for sodium. Beyond 48 h of malting, mineral content increased, particularly, for 96 h in FM grain malt. Significant (p < 0.05) increases in the mineral content of FM varieties were noted at 48 h and 96 h of malting. Increase occurred at 72 h of malting for potassium, iron and boron. Malting did not have any effect on the manganese content of the dark brown FM; however, it increased the manganese content at 48 h of malting for brown FM. Malting for 96 h significantly (p < 0.05) reduced sodium content. Consecutive decrease in phytic acid content of the grains was not recorded with durations in malting time. Although statistically significant differences (p < 0.05) were observed, malting did not result in too much change in the physicochemical properties of the grains. Several flavonoids, catechin, epicatechin, quercetin, taxifolin, and hesperitin were isolated, whilst protocatechuic acid was the only phenolic acid detected in the unmalted and malted FM. Increases in catechin, epicatechin and protocatechuic acid were observed for 72 and 96 h malt of brown FM with similar observations recorded for sorghum. Complete loss of taxifolin, catechin, and hesperitin were noted with malting time. FM grains exhibited 2, 2-diphenyl-1-picrylhydrazyl (DPPH), 2, 2՛-azinobis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging action and iron reducing activities. Increased iron reducing activity alongside ABTS radical scavenging activity was recorded with malting time. A fermentation-time dependent decrease in the pH of the non-alcoholic beverages, with a corresponding increase in sugar content were recorded. A similar decrease was also recorded for the viscosities of the beverages. The FM malt beverages were found to contain a higher amount of citric acid compared to the sorghum malt beverage. A decrease in the citric acid content with fermentation time was noted in the grain malt beverages fermented with Lactobacillus fermentum, particularly for the iv FM beverage. The phenolic compounds detected in the FM malt beverages fermented with the grain microbial flora and Lactobacillus fermentum were protocatechuic acid, catechin and epicatechin. Taxifolin and kaempferol along with the earlier mentioned compounds were detected in the sorghum malt beverage. Catechin was found in higher amount compared to other phenolic compounds in the FM and sorghum malt beverages. FM malt beverages were found to contain a higher amount of total phenolics compared to the beverage prepared from sorghum malt. Fermentation with the grains’ microbial flora and L. fermentum resulted in reduced total phenolics of FM and sorghum malt beverages, particularly after 24 h of fermentation. A fermentation-time dependent decrease in total phenolics of FM beverages fermented with L. fermentum was noted. Fermentation within 24 - 48 h using the grain microbial flora showed higher total individual phenolic compounds for the dark brown FM and sorghum, compared to other fermentation periods. Fermentation of the beverages for 24 h retained a higher amount of the total phenolics compared to other fermentation periods, especially for the L. fermentum beverages. Reduced total phenolic content and antioxidant activity of the beverages were noted at 24 h of fermentation for the two microbial sources. Significant (p < 0.05) increases in total phenolics were observed within 72 – 96 h of fermentation of the brown FM malt beverage with the grains’ microbial flora. Fermentation for 72 h and 96 h with L. fermentum increased the total phenolic content of the brown FM. Increase in total flavonoid content (TFC) of brown FM malt beverage was noted at 72 h fermentation for both microbial sources. Unlike with L. fermentum, no significant (p > 0.05) change in TFC was observed for the dark brown FM beverage after 24 h fermentation with the grains’ microbial flora. Beverages exhibited DPPH, ABTS radical scavenging action and iron reducing activities, which were significantly (p < 0.05) reduced at 96 h fermentation for both microbial sources. The 24 h fermented beverage retained a higher amount of total phenolic and flavonoid contents, and had higher antioxidant activity compared to other fermentation periods for both microbial sources. The study shows that FM is a rich source of essential minerals and v phenolic compounds, and demonstrates that 72 to 96 h of malting has a positive effect on minerals and certain phenolic compounds over the 48 h malting period widely used for preparation of FM malt. The presence of hesperitin in FM grain was established. A new method was developed for the production of FM non-alcoholic beverage with measurable amounts of health-promoting compounds. An ideal fermentation period (24 h) for FM malt non-alcoholic beverage production with enhanced health-promoting compounds, using Lactobacillus fermentum was demonstrated. Fermentation limit (96 h) for production of FM malt beverage using either the grain microbial flora or L. fermentum was confirmed. These findings provide a rationale for increased utilisation of FM as a functional food grain, and its use as malt in production of non-alcoholic beverage for health promotion and wellness.
NRF

submitted in fulfilment of the academic requirements for the degree of Master of Applied Science in Food Science and Technology, Durban University of Technology, 2015.
Vitamin A deficiency (VAD) is a major health problem in sub-Saharan Africa where maize is a staple food. Amahewu, a fermented non-alcoholic,maize-based beverage is a popular drink in southern Africa.The aim of this study is to produce a provitamin A enriched and acceptable amahewu, using provitamin A biofortified maize which can be used to alleviate VAD. The optimal processing parameters for the production of amahewu using provitamin A-biofortified maize were determined. Amahewu samples were prepared with reference to a traditional method by boiling a mixture of maize meal and water (rato:1:7) at 90ᴼC, with occasional stirring, for 15 minutes. The resulting porridge was left to cool to approximately 40ᴼC, before inoculation and fermentation at 37oC. Processing parameters investigated were inoculum types (wheat bran (WB), maize malt (MM) and Lactobacillus mixed starter culture) and inoculum concentration (0.5,1 and 2% (w/w)) and varieties of provitamin A maize (PVAH 62 and PVAH 19). Wheat flour (at 2%) was used as reference inoculum to conform to the traditional practice. White maize amahewu samples processed in the same way as those of provitamin A-biofortified maize were used as references. Provitamin A amahewu samples were produced using the optimized processing parameters and then analysed for nutrient composition, including carotenoids, protein, ash, amino acids, mineral profile and invitro protein digestibility. The consumer acceptability of amahewu samples was evaluated using regular consumers of amahewu (n= 54), who rated the acceptability of the samples on a 9-point hedonic scale (1:disliked extremely, 9:liked extremely). The storage stability of the provitamin A biofortified amahewu samples was assessed by subjecting the samples to different storage conditions: 4ᴼC, 25ᴼC and 37ᴼC. The microbiological quality of the stored samples was monitored by taking samples every day for a period of five days to analyse for the presence of aerobic and anaerobic bacterial spore formers, E.coli and moulds. The provitamin A maize variety did not influence pH and Total titratable acidity (TTA) of amahewu samples during fermentation. As expected, there was a substantial drop in pH with fermentation time. After 24 hours, all the samples of amahewu, including those made with white maize, prepared using malted maize and wheat bran inocula reached a pH of 3.3-3.8 and TTA of 0.3-0.6, which were within acceptable range for amahewu. The addition of a starter culture substantially reduced fermentation time, from 24 to six hours. The inoculum of WB and MM, respectively, at a concentration of 0.5%, with or without starter culture (5%), were found to be suitable for the production of amahewu using provitamin A biofortified maize. The total provitamin A content of amahewu samples, produced using optimised parameters (i.e one variety of provitamin A biofortified maize, 0.5% MM, WB with or without starter culture), ranged from 3.3-3.8 μg/g (DW). The percentage retention of total provitamin A ranged from 79%- 90% (DW). The lowest percentage retention was observed in products fermented with the addition of starter culture. The gross energy of the amahewu samples was approx. 20 MJ/kg. There was a slight increase in the lysine content of amahewu after fermentation. The protein digestibility (approx. 91%) of amahewu samples was slightly higher than that of raw provitamin A maize (86%). Amahewu processed using starter cultures had a slightly higher iron content than those processed without a starter culture. Consumer acceptability data showed that amahewu samples made with provitamin A biofortified maize were slightly more acceptable (average rating for overall acceptability was 7.0 ± 1.2), compared to those made with white maize (average rating for overall acceptability was 6.4 ± 0.8). Principal component analysis (PCA) of Amahewu sensory data showed that 71% of variation was due to maize types and 18% of variation may be due to the inoculum used during fermentation. The use of a starter culture improves the taste and aroma acceptability of amahewu. Segmentation of consumers based on overall linking for amahewu revealed three clusters, named A, B and C. Cluster A consisted of most consumers (43%), who liked amahewu moderately. About 60% of these consumers were females. Cluster B consisted of most of the consumers (31%) who were undecided about their liking for the product. Approximately 52% of the consumers in this cluster were female. Cluster C consisted of consumers (26%) who liked amahewu very much. Sixty-four percent (64%) of these consumers were female. It appeared that gender may have some influence on overall liking for amahewu, as cluster B, consisting of undecided consumers, had more male consumers compared to clusters A and C. Age did not seem to be significantly associated with the liking of amahewu. Provitamin A biofortified amahewu samples stored under refrigerated conditions (4ᴼC) had better microbiological quality compared to those stored at 25ᴼC and 37ᴼC. Refrigeration effectively maintains the microbiological quality of amahewu for about three of days. Provitamin A biofortified maize can be used to produce β-carotene enriched amahewu that is acceptable to consumers following the processing method that is traditionally employed for white amahewu at both domestic and commercial level. Provitamin A biofortified amahewu has the potential to make a significant contribution towards alleviating VAD among rural communities, who are the most vulnerable to VAD.