Thesis (MSc Food Sc)--Stellenbosch University, 2002. / ENGLISH ABSTRACT: The start-up period of Upflow Anaerobic Sludge Bed (UASB) reactors can significantly be reduced by enhancing the time-consuming granulation process through the batch cultivation of anaerobic granular sludge and thus seeding the reactor with this cultivated granular sludge, instead of raw anaerobic sludge. The precise mechanism for granule formation is not well known, but it is believed that extracellular polymers (ECP) play a critical role in the granulation process. Information on the precise role of ECP is also limited and no universal standardised method for ECP extraction is used at present. Therefore, comparison of results from different researchers has to be made with great caution. The objectives of this study were to evaluate an ECP extraction methód so as to optimise the extraction time, and then to correlate ECP composition of UASB granules with granule metabolic activity. The impact of changes in the environmental conditions, such as sludge sources (Paarl and Kraaifontein-sludge), carbon growth substrates (yeast extract lactate, glucose medium and fruit cocktail effluent) and batch cultivation techniques (roller-table and shake-waterbath), on batch cultivation studies was also evaluated in terms of granule activity, ECP composition and granule formation. A physical extraction method was used to quantify the ECP content of UASB granules from six different sources. The optimal extraction time was taken as the time needed before cell lysis took place, and before intracellular material started contributing to the ECP content of the granules. It was concluded that the ECP composition was affected by the wastewater composition fed to the original UASB reactors, It was also found that the activity test results could be used to indirectly predict the activity of the different trophic groups present in the UASB granules. A correlation was found between the activity test results and the total ECP content, and this showed that the granules with the higher ECP yields exhibited greater biogas (SB) and methanogenic (SM) activities. However, based on the activity data and total ECP content, it appeared that a protein:carbohydrate ratio < 1 affected the activity of the granules, The sludge source used as inoculum for batch cultivation of anaerobic granular sludge had a significant effect on granule formation. The use of a pre- granulated raw anaerobic sludge, such as the Paarl-sludge, resulted in a greater increase in granule numbers at the end of the cultivation period. The acetic acid activity profiles showed that the acetoclastic methanogens that are involved in initiation of granulation by nucleus formation, were inactive or absent in the different batch systems, with the exception of the roller-table glucose cultivated Kraaifontein-sludge (RKG) batch system. The addition of glucose as carbon growth substrate for batch cultivation not only enhanced the activity of the acidogenic population, but also led to the establishment to a greater variety of granule trophic groups within all the glucose cultivated batch systems. The addition of fruit cocktail effluent as carbon substrate enhanced ECP production in the Paarl-sludge cultivated batch systems. However, the addition of carbon substrates showed no discernible trend on granule formation itself. The roller-table cultivation technique resulted in the higher increase in granule numbers, and it was speculated that the more vigorous shake-waterbath technique probably shortened the contact time between biomass and substrate. Large variations in the ECP composition of the different batch systems were found, and these were ascribed to the composition heterogeneity of different sludges. For future studies, it is advisable to characterise sludge, both chemically and microbiologically before using as inoculum. The selection of an appropriate sludge inoculum should then lead to optimisation of the granulation process. / AFRIKAANSE OPSOMMING: Die aanvangsperiode van "Upflow Anaerobic Sludge Bed" (UASB) bioreaktors kan noemenswaardig verminder word deur die tydsame granulasie proses te versnel deur die vooraf lot-kweking van anaërobe granulêre slyk waarmee 'n reaktor dus geïnokuleer kan word in plaas van rou anaërobe slyk. Die presiese meganisme van granulevorming is nog nie welbekend nie, maar daar word beweer dat ekstrasellulêre polimere (ECP) wel 'n kritiese rol speel in die granulasie proses. Inligting; oor die presiese rol van ECP is ook nog beperk, en tans word daar nog , geen universele standaard metode vir ECP-ekstraksie gebruik nie. Gevolglik moet resultate vanaf verskeie navorsers met groot omsigtigheid vergelyk word. Die doelwitte van hierdie studie was om 'n ECP ekstraksiemetode te evalueer deur die ekstraksietyd te optimiseer, en dan te korreleer met die ECP samestelling en metaboliese aktiwiteit van die UASB granules. Die inwerking van veranderinge in omgewingskondisies, soos slykbronne (Paarl- en Kraaifontein-slyk), koolstofbronne (gisekstrak-Iaktaat-, glukose-medium en vrugtekelkie-uitvloeisel) en lot-kwekingstegnieke (rol-tafel en skud-waterbad) op lot-kweking studies in terme van granule aktiwiteit, ECP-samestelling en granulevorming is ook ondersoek. 'n Fisiese ekstraksie metode is gebruik om die ECP-inhoud van UASB granules vanaf ses verskillende bronne te bepaal. Die optimale ekstraksietyd is geneem as die tyd benodig voordat sellise sal plaasvind en die intrasellulêre materiaal 'n bydrae sal lewer tot die ECP-inhoud van granules. Dit is afgelei dat die ECP-samestelling beïnvloed word deur die samestelling van die afvalwater wat vir die oorspronklike UASB bioreaktors gevoer is. Voorts is gevind dat die aktiwiteitstoets resultate indirek gebruik kan word vir die voorspelling van aktiwiteit van die verskillende trofiese groepe wat in die UASB granules teenwoordig is. 'n Korrelasie is gevind tussen die aktiwiteitstoets resultate en die totale ECP-inhoud wat aangedui het dat granules met hoër ECP opbrengste, beter biogas (SB) en metanogeniese (SM) aktiwiteit getoon het. Volgens die aktiwiteitsdata en totale ECP-inhoud het dit egter geblyk dat 'n proteïen:koolhidraat verhouding < 1 die aktiwiteit van granules beïnvloed het. Die slykbron wat as inokulum gebruik is vir lot-kweking van' anaërobiese granulêre slyk het 'n noemenswaardige invloed gehad op granulevorming. Die gebruik van 'n rou anaërobe slyk wat reeds 'n mate van granulasie getoon het, soos die Paarl-slyk, het 'n hoër toename in granule getalle aan die einde van die kwekingsperiode teweeg gebring. Die asynsuur aktiwiteitsprofiele het aangedui dat die asetoklastiese metanogene, wat hoofsaaklik betrokke is by inisiëring van granulasie deur kernvorming, onaktief of afwesig was in al die verskillende lotsisteme, met die uitsondering van die roltafel glukose-gekultiveerde Kraaifonteinslyk (RKG) lot-sisteem. Die toevoeging van glukose as koolstofbron vir lot-kweking het nie alleenlik die aktiwiteit van die asidogene populasie verhoog nie, maar het ook bygedra tot die vestiging van 'n groter verskeidenheid van granule trofiese groepe. Die toevoeging van vrugtekelkie-uitvloeisel as koolstofbron het die produksie van ECP verhoog in die Paarl-slyk gekweekte lot-sisteme. Die toevoeging van koolstofsubstrate het egter geen merkbare verandering getoon in granulasie opsigself nie. Die rol-tafel kwekingstegniek het 'n hoër toename in granule getalle tot gevolg gehad, en dit is gespekuleer dat die meer kragtige skud-waterbad tegniek waarskynlik die kontaktyd tussen die substraat en biomassa verkort het. Groot variasies is gevind in die ECP-samestelling van die verskillende lotsisteme, en dit is toegeskryf aan die heterogene samestelling van die verskillende slyke. Vir toekomstige navorsing is dit raadsaam om slyk voor gebruik as inokulum beide chemies en mikrobiologies te karakteriseer. Die keuse van 'n geskikte slyk-inokulum sal bydra tot die optimisering van die granulasie proses.

Thesis (MSc Food Sc (Food Science))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Over the last decade, studies on the quality of the water in many of South Africa.s rivers revealed an increase in pollution levels (DWAF, 1996; DWAF, 2000). In agricultural areas of the Western Cape many farmers draw water for irrigation from nearby rivers without knowing what the microbial pollution level is. In order to investigate the current quality of river water in some of the Western Cape.s major rivers and to explore the possibility of pathogen carryover from water onto produce, this research project was initiated. In an exploratory study over a five month period, the microbiological and water chemistry of three selected sites from the upper Berg and two from the Plankenbrug Rivers were assessed. Studied organisms included Salmonella, Staphylococcus, Listeria, endosporeformers, coliforms, Escherichia coli and intestinal Enterococci, while the chemical analysis consisted of pH, alkalinity, conductivity and chemical oxygen demand (COD). Faecal coliform counts ranging from 540 to 1 700 000 cfu.100ml-1 and 490 to 160 000 cfu.100ml-1 were found for the Berg and Plankenbrug Rivers, respectively. The water temperature ranged from 12º to 21ºC with COD values always below 100 mg.L-1. Potential human pathogens such as Salmonella, Staphylococcus, Listeria, endosporeformers, E. coli and intestinal Enterococci were frequently isolated from all five sites that were sampled. These results are of great concern to farmers, fresh produce retailers and consumers alike as the river water is regularly drawn for irrigation of produce that is later consumed raw or after a minimal processing step. From the exploratory study it was concluded that the water from all the sites were not suitable for use in irrigation practices as they regularly exceeded the guidelines for faecal coliforms and E.coli as set out by South African authorities. Irrigation with faecally polluted river water is one way that fruit and vegetables can become contaminated with foodborne pathogens. The risk of disease transmission from potential pathogens present in the irrigation water is influenced by the microbial load present and the numbers carried over to the produce. In this study the carry-over and survival of Escherichia coli on green beans, sugar-snap peas and cocktail tomatoes was assessed under controlled laboratory conditions. The produce was exposed to E.coli under different combinations of exposure times (5, 15 or 30 min), drying times (30 or 120 min) and different inoculum concentration ranges (107, 105, 104, 10³ and 10ª). In all cases a reduction of at least one log value in original inoculum number was found with the 10ª inoculums showing no survivors. Follow-up studies with an exposure time of 60 min and increased drying times of 6 or 12 h. Neither of these parameter changes affected the variation in numbers for the same inoculum or the E.coli survivors. Similar E.coli loads to those detected on the green beans were detected on the sugar-snap peas. The number of survivors on the cocktail tomatoes was much lower than found for the beans and peas. The number of survivors on the cocktail tomatoes was much lower than found for the beans and peas. This was attributed the 'smooth'surface of the tomato skin probably making attachment of the E.coli bacteria difficult. With the exception of the 10ª inoculum range, all E.coli survivors detected on the three types of produce studied exceeded the guideline numbers set for fresh produce. If similar survival patterns are to be found in the environment then results from this study should serve as a warning that the Plankenburg river water is unsafe for use in the irrigation of fresh produce. Some farmers are already treating river water with chlorine prior to irrigation to eliminate the chance of pathogen transfer onto produce. However, this is not a feasible solution for most farmers due to the high cost of implementing a system such as this. / AFRIKAANSE OPSOMMING: Oor die afgelope tien jaar het studies oor die gehalte van die water in baie van Suid-Afrika se riviere 'n toename in besoedelingsvlakke getoon. Baie boere in landbou gebiede van die Wes-Kaap wat watertrek vir besproeiing uit die nabygelee riviere is onbewus van die mikrobiese besoedelingsvlak van die rivier. Hierdie navorsingsprojek is geinisieer ten einde die huidige gehalte van die water in sommige van die Wes-Kaap se riviere te monitor, asook die moontlikheid van patogeen oordrag van die water na vars produkte te ondersoek. Met behulp van 'n verkennende studie oor . vyf maande tydperk, is die mikrobiologiese en water chemie kenmerke van drie geselekteerde moniteringspunte uit die boonste Bergrivier, en twee uit die Plankenburgrivier bemonster. Die bestudeerde organismes het ingesluit: Salmonella, Staphylococcus, Listeria, endosporeformers, fekale kolivorme, Escherichia coli en intestinale Enterococci. Die chemiese parameters het bestaan uit pH, alkaliniteit, geleiding, temperatuur en chemiese suurstof behoeftes (CSB). Fekale kolivorme tellings het gewissel van 540 - 1 700 000 cfu.100ml-1 en 490 tot 160 000 cfu.100ml-1 is gevind vir die Berg- en Plankenburgriviere, onderskeidelik. Die water temperatuur het gewissel van 12 º tot 21ºC met die CSB-waarde altyd onder 100 mg.L-1. Potensiele menslike patogene soos Salmonella, Staphylococcus, Listeria, endosporeformers, E. coli en intestinale Enterococci is dikwels geisoleer uit al vyf die moniteringspunte. Hierdie resultate is van groot belang vir boere, kleinhandelaars en varsprodukte verbruikers omdat die riviere se water gereeld onttrek word vir besproeiing van produkte wat later gebruik word in rou of in minimale geprosesseerde vorm. Vanuit die verkennende en uit die langer termyn studie (12 maande) is die gevolgtrekking gemaak dat die water vanuit die riviere nie geskik is vir gebruik in besproeiingspraktyke waar die riglyne vir fekale kolivorme en E. coli, soos uiteengesit deur die Suid-Afrikaanse owerhede, gereeld oorskry word nie. Besproeiing met fekale besoedelde rivierwater is een manier waarop groente en vrugte besmet kan word met voedsel patogene. Die risiko vir die oordrag van moontlike patogene teenwoordig in die besproeiingswater word beinvloed deur die mikrobiese lading teenwoordig en die getalle wat oorgedra word na die produkte. In hierdie studie is die oordrag en oorlewing van E. coli op groenboontjies, ertjies en tamaties onder beheerde laboratoriumtoestande beoordeel. Die produkte is blootgestel aan E. coli onder verskillende kombinasies van blootstellingstydperke (5, 15 of 30 min), drogingstye (30 of 120 min) en verskillende inokulum konsentrasies (107, 105, 104, 10³ en 10ª). In elke geval is . afname van ten minste een log waarde van die oorspronklike inokulum konsentrasie gevind. Die 10ª inokulum reeks het geen oorlewende selle getoon nie. Opvolg studies is gedoen met 'n blootstellingstyd van 60 min en verlengde drogingstye van 6 of 12 uur. Daar is gevind dat hierdie parameter veranderings die variasie in getalle vir dieselfde inokulum of die oorlewende E. coli nie beinvloed het nie. Soortgelyke E. coli ladings wat op die groenbone aangetref is, is ook op die ertjies aangetref. Die aantal oorlewendes op die tamaties was heelwat laer as die resultate vir die boontjies en ertjies. Dit word toegeskryf aan die "gladde" oppervlak van die tamatie-vel wat waarskynlik die aanhegting van die E. coli selle bemoeilik. Met die uitsondering van die 10ª inokulum reeks, het al die E. coli oorlewende selle gevind op die drie tipes produkte, die riglyn getalle oorskry wat vir vars produkte gestel is. As soortgelyke oorlewingspatrone gevind word in die omgewing, behoort die resultate van hierdie studie as waarskuwing te dien dat die water van die Plankenburgrivier ongeskik is vir die besproeiing van vars produkte. Sommige boere is reeds besig met chloor behandeling van die rivierwater voor besproeiing, om die kans van patogeenoordrag na die produkte uit te skakel. Dit is egter nie 'n haalbare oplossing vir die meeste boere nie as gevolg van die hoe kostes verbonde aan die implementering van' n stelsel soos hierdie.

Thesis (MSc Food Sc )--Stellenbosch University, 2003. / ENGLISH ABSTRACT: This study shows the potential of near infrared spectroscopy for both qualitative and quantitative analyses of wine and brandy in the distilling industry. Wines intended for further processing by the distillation industry have to meet certain specifications to ensure an efficient and cost-effective distilling process with a suitable product yield. Certain compounds have, therefore, been recognised as significant quality parameters and through their regulation, a product of consistent quality can be ensured. These quality parameters include alcohol, total acid, volatile acid, total sulphur dioxide, total phenolics, reducing sugars and acetaldehyde concentrations and pH. The effective control of all these compounds in the wines prior to distillation, is critical to maintain the feasibility and efficiency of the process. Fourier transform near infrared spectroscopy (FT-NIRS) can be used as a rapid and accurate analytical method for the determination of alcohol concentrations in distilling and brandy base wine. The choice of path length for the liquid sampling cell is an important consideration. In this study, alcohol with its strong absorption of OH bands in the near infrared region was not significantly affected, but difference in path length showed a significant influence on the prediction of the volatile acid concentration in distilling wine. Very strong correlations were found between the spectral data and the alcohol concentration data in distilling wine (r = 0.99, SEP = 0.18% v/v, RER =30) and brandy base wine (r = 0.92, SEP = 0.18% v/v, RER = 11.1). Good predictions were obtained for the volatile acid content of brandy base wine (r = 0.85, SEP = 0.04 g.L-1 ) but better accuracy could be obtained by incorporating a wider concentration range and more sensitive, although not the accepted reference methods, such as HPLC and GC, into the modelling. In contrast to the brandy base wine, the much cruder, turbid distilling wine (r = 0.67, SEP = 0.33 g.L-1 ) yielded poor calibration results for volatile acid. FT-NIRS can also be used as a rapid screening method to measure the total acid content, pH and total phenol levels in brandy base wine samples. Satisfactory predictions were obtained for the total acid content (r = 0.89, SECV = 0.38 g.L-1 ), pH (r = 0.84, SEP = 0.09) and total phenol levels (r = 0.71, SEP = 16.4 mq.L" GAE). The SEP of all the parameters compared well with the SEL and were within acceptable limits. The range of the concentration measurements were, however, very narrow and the accuracy of these models (measured as the ratio of the range to the standard error of prediction) were all below 10, indicating that modelling of these parameters in brandy base wine should be attempted with reference values covering a wider range. Poor correlations and predictions were obtained between the wine spectral data and the acetaldehyde (r = 0.39 , SECV =1.45 rnq.L"), residual sugar (r = 0.58, SECV =0.49 mq.L" ) and total sulphur dioxide (r = 0.35, SEP = 10.9 mq.L" ) concentration data. The unique character of potstill distillates, originating from the grape cultivar, type and age of the oak casks, lends itself to the development and blending of different brandies. The process of blending and correcting the brandy is necessary to obtain a product and brand of established and recognised character. The brandy is generally divided into different classes prior to blending based on the degree of "smoothness" or "hardness" of the matured distillate as determined by a sensorial panel. The ideal would, however, be to replace this subjective evaluation method with a more objective approach, such as an instrumental method like NIRS. Near infrared spectroscopy showed potential to successfully discriminate and classify brandy samples in terms of their sensorial classification status. The near infrared spectra of unblended three-year old brandies were measured and discriminant analysis performed using the subjective sensory classification data of the brandy. SIMCA (soft independent modelling of class analogy) models were developed by calculating separate peA (principal component analysis) models for each of the classes for which qualitative analysis were required. An assessment of class membership of an unknown sample was made on the basis of the distance to the cluster centroid. Two types of NIRS instruments, a Fourier transform near infrared system and a diode array spectrophotometer were employed to examine the discriminatory abilities of NIRS. Excellent predictions (90.9%-100%) were obtained for the "smoothest" and "hardest" classes within the 2000 and 2001 seasons with FT-NIRS. For the diode array spectra reasonable classification data were obtained for all the classes within the 2000 and 2001 seasons, as well as an excellent classification rate (100%) of the smoothest class in the 1999 season. Prediction results for class membership of the brandy samples from the three season's combined data did not deliver satisfactory discriminatory results. By applying NIRS and SIMCA, rapid and objective classification of brandy samples within seasonal context can be established to enhance the blending processes. Having accurate reference data in the calibration set is essential in near infrared spectroscopy (NIRS). The precision (usually decided by blind duplicate determination) of the reference data serves as the performance target for correlation-based near infrared analysis. The differences between reference values obtained from two independent laboratories and the Foss Winescan FT 120, which is also a predictive method, for the alcohol and volatile acid determinations in distilling wine and between two independent laboratories for the measurement of the total sulphur dioxide content, were investigated. ANOVA and post-hoc analysis (Bonferroni testing) revealed statistically significant differences (p<O.OS)between all three measurements for the volatile acid analysis. One laboratory differed significantly from the other two measurements in its measurement of the alcohol content in distilling wine. A paired t-test performed on the two laboratories' values obtained for total sulphur dioxide revealed statistically significant differences (p<O.OS) between the two sets of results. This study restated the importance of obtaining reliable reference data for calibration purposes to maintain the efficiency and accuracy and therefore the reputability of NIRS as an accurate and reliable analytical method. / AFRIKAANSE OPSOMMING: Hierdie studie dui op die potensiaal van naby infrarooi spektroskopie (NIRS) vir beide die kwalitatiewe en kwantitatiewe analise van wyn en brandewyn in die distillasie industrie. Sekere spesifikasies word gestel waaraan wyn met voldoen voor dit verdere prosessering tydens distillasie kan ondergaan. Die regulering van wynkwaliteit is noodsaaklik om die distilleringsproses ekonomies volhoubaar te maak. Sekere komponente in die wyn het 'n direkte invloed op die kwaliteit van die eindproduk en word beskou as betekenisvolle parameters wat beheer moet word om 'n produk van konstante hoë kwaliteit te lewer. Hierdie parameters sluit alkohol, titreerbare suur- en vlugtige suurinhoud, asook die pH, totale swaweidioksied, totale fenol vlakke, reduserende suikers en asetaldehied konsentrasies in. Die regulering van al hierdie komponente in die wyne is essensieel voor die aanvang van distillasie om die effektiwiteit en produktiwiteit van die proses te verseker. Fourier transformasie naby-infrarooi spektroskopie (FT-NIRS) kan benut word as 'n vinnige en akkurate analitiese metode om die alkohol konsentrasie in stook- en rabatwyn te bepaal. 'n Geskikte padlengte kuvet moet gekies word vir die ontleding van vloeistof monsters met NIRS. In hierdie studie is gevind dat die voorspelling van alkoholkonsentrasie in wyn, nie soseer beïnvloed is deur die keuse van padlengte nie. 'n Vlugtige komponent soos asynsuur, wat die maatstaf van die vlugtige suurinhoud van die wyn is, word egter regstreeks beïnvloed deur die keuse van padlengte. 'n Sterk korrelasie is gevind in die datastel vir die alkoholinhoud van die stookwyn (r = 0.99, SEP = 0.18% v/v, RER = 30) en rabatwyn (r = 0.92, SECV = 0.18% v/v, RER = 11.1). Baie goeie voorspellings is ook gevind vir die vlugtige suurinhoud van rabatwyn (r = 0.85, SEP = 0.04 g.L-1). Die akkuraatheid van die metode (uitgedruk as die RER waarde) het egter aangedui dat 'n wyer konsentrasie omvang en meer akkurate verwysingswaardes gebruik moet word om 'n geskikte model vir vlugtige suur kwantifikasie in rabatwyn te bewerkstellig. In teenstelling met die resultate verkry vir die rabatwyn, het die ruwer, turbiede stookwyn swak gekalibreer vir die vlugtige suurinhoud (r = 0.67, SEP = 0.33 g.L-1 ). FT-NIRS kan ook gebruik word as 'n vinnige seleksie metode om titreerbare suur vlakke, pH en totale fenolinhoud van rabatwyn monsters te bepaal. Sterk korrelasies is in die datastel gevind vir die titreerbare suurinhoud (r = 0.89, SECV = 0.38 g.L-1 ), pH (r = 0.84, SEP = 0.09) en totale fenol vlakke (r = 0.71, SEP = 16.4 rnq.L" GAE) van rabatwyn. Die SEP waardes verkry vanaf die NIRS bepalings het baie goed vergelyk met die standaard foute wat verkry is vir die chemiese bepalings. Die konsentrasie omvang van die verwysingswaardes was, egter, beperk en die akkuraatheid van die NIRS modelle (uitgedruk as die verhouding tussen die konsentrasie omvang en die standaard fout van voorspelling) was laer as 10. Dit het daarop gedui dat die NIRS kalibrasie van die parameters eerder uitgevoer moet word met monsters wat 'n wyer konsentrasie omvang dek en gemeet is met meer sensitiewe analitiese metodes soos HPLC en GC. Die kalibrasie en voorspelling van die asetaldehied inhoud (r = 0.39 , SECV =1.45 rnq.L"), reduserende suiker konsentrasie (r = 0.58, SECV =0.49 mq.L" ) en totale swaweldioksiedinhoud (r = 0.35, SEP = 10.9 rnq.L" ) het onaanvaarbare voorspellings gelewer. Die unieke karakter van potketel distillate wat bepaal word deur die druifkultivar, tipe en ouderdom van die eikehoutvate, bied die potensiaal vir die ontwikkeling en versnyding van verskillende style brandewyn. In 'n poging om 'n produk en handelsnaam van erkende en gevestigde karakter te verkry, moet die brandewyn eers versny word na veroudering. Die brandewyn word geklassifiseer in verskillende style op grond van sensoriese gehalte. NIRS is gebruik as 'n vinnige tegniek om te onderskei tussen verskillende style onversnyde drie jaar-oue brandewyn. SIMCA (soft independent modelling of class analogy) modelle is ontwerp deur aparte PCA (hoof komponent analise) modelle vir elke klas waarvoor kwalitatiewe analise vereis is, te ontwikkel. 'n Skating van klas lidmaatskap van 'n onbekende monster is gemaak deur die afstand van die monster na die groepsmiddelpunt te bereken. Twee soorte NIRS instrumente, 'n FT-NIR en 'n diode array spektrofotometer is gebruik tydens die studie om die diskrimineringsvermoë van elk te ondersoek. Uitstekende voorspellings (90.9%-100%) is gekry vir die "sagste" en "hardste" klasse binne die 2000 en 2001 seisoene met FTStellenbosch NIRS. Met die diode array spektra is redelike klassifikasie resultate verkry vir al die klasse binne die 2000 en 2001 seisoene, asook 'n uitstekende klassifikasie (100%) vir die sagste klas binne die 1999 seisoen. Voorspellings vir klas lidmaatskap van die brandewyn monsters vir die drie seisoene se gekombineerde data het egter onaanvaarbare diskriminasie resultate gelewer. NIRS en SIMCA sal 'n vinnige en objektiewe klassifikasie proses van onversnyde brandewyn monsters kan meebring wat tot voordeel van die versnydings- en produksieproses sal wees. Hierdie voorspellings sal egter binne seisoensverband moet geskied. Akkurate verwysingsdata in die kalibrasie datastel is 'n essensiële fasset van naby infrarooi spektroskopie. Die presisie (bepaal deur blinde duplikaat bepalngs) van die verwysingsdata dien as 'n prestasie teiken vir korrelasie-gebaseerde naby infrarooi voorspellings. Die verskille tussen die data verkry vir die alkohol- en vlugtige suurinhoud van stookwyn deur twee onafhanklike laboratoriums en 'n instrumentele metode (Foss Winescan FT 120) asook die verskille tussen die data verkry van twee laboratoriums vir die totale swaweidioksied metings, is ondersoek. ANOVA en post-hoc toetse het aangedui dat al die data verkry vir die vlugtige suur bepalings, statisties betekenisvol van mekaar verskil het (p<0.05). Vir die alkohol bepalings in stookwyn, het die data verkry van een laboratorium betekensvol verskil van die ander twee datastelle. Gepaarde t-toetse uitgevoer op die data verkry vir totale swaweldioksiedinhoud, het ook betekinisvolle verskille aangedui tussen die twee laboratoriums. Hierdie studie het die belangrikheid van betroubare verwysingsdata vir kalibrasie doeleindes beklemtoon, veral waar 'n tegniek soos NIRS se werksverrigting afhang van die akkuraatheid van dié waardes.

Thesis (MSc Food Sc )--Stellenbosch University, 2003. / ENGLISH ABSTRACT: The effect of drying conditions on the retention of harpagoside, a glucoside iridoid found in the indigenous Southern African plant Harpagophytum procumbens (Devil's Claw), was investigated. Additionally, the feasibility of using near infrared spectroscopy (NIRS) as a rapid and nondestructive analytical tool for the prediction of the iridoids harpagoside (HS), 8-p-coumaroyl harpagide (8pCHG) and moisture contents, as well as the HS:8pCHG ratio in Devil's Claw was also studied. Due to the purported medicinal activity ofthe tuberous secondary roots, Devil's Claw is of commercial interest with a well established export market in Germany. The stability of HS during processing has, however, never been investigated and the current study evaluated the effect of three drying methods (sun, tunnel and freeze-drying), as well as three tunnel-drying temperatures (40°C, 50°C and 60°C, dry bulb temperature) on HS content. Methanolic extracts (70% methanol-water) were analysed by high-performance liquid chromatography (HPLC) and a multiple extraction test (n = 12) indicated a 91.70% recovery for HS. The comparative drying results showed significant (P < 0.05) differences between tunnel and freeze drying on the one hand and sun-drying on the other, with the latter being the most detrimental technique. In comparing tunnel-drying temperatures, at a constant relative humidity (RH) of30%, drying at 40°C showed the lowest retention ofHS, differing significantly (P < 0.05) from that at 50°C. Drying at 60°C did not differ significantly (P > 0.05) from either of the other two conditions, but also resulted in lower retentions ofHS content than drying at 50°C. Overall, freeze-drying showed excellent results for HS retention (1.565% ± 0.394%; mean ± standard deviation), but economic constraints limit the commercial application of this drying method. Although further study is required to optimise tunnel-drying conditions, this method indicated acceptable and practical results (1.526% ± 0.396%), especially with conditions set to 50°C and 30% RH (1.750% ± 0.438%). Future research should concentrate on elucidating the degradation mechanisms of HS during processing, transport and storage of dried Devil's Claw root. The analysis of dried, ground Devil's Claw root by NIRS was referenced to HPLC analyses of the same methanolic extracts used in the drying studies. The spectra were generated on two different NIRS instruments by measuring diffuse reflectance of the powder whilst calibrations were performed by partial least squares regression. Both independent data set validation and full cross-validation were used to evaluate the performance and predictive abilities of the various models. Excellent moisture content (MC) calibrations were developed with a standard error of prediction (SEP) ofO.24% (range: 2.44% to 10.43%) and correlation coefficient (r) of 0.99. For the given MC range, the model performed very well against the standard error of laboratory (SEL) ofO.14%. Considering the sensitivity of the HPLC reference method (SEL = 0.035%), the HS content calibration (range: 0.693% to 2.244%) performed admirably with an SEP ofO.134%. The available range was, however, relatively small and future NIRS studies should incorporate a larger range if possible. The 8pCHG model (range: 0.069% to 0.290%) performed similarly, with a standard error of cross-validation of 0.028% and SEL of 0.007%. By inclusion of a second species, H. zeyheri, both the 8pCHG content and especially HS:8pCHG ratio (range: 1.84 to 34.48) calibrations were evaluated as possible methods of distinguishing between species. With 8pCHG values of ca. 0.9%, good separation was achieved for the first model but, contrary to previous studies, the ratio was found to be less successful. Principle component analysis of the spectra, however, showed great potential as a qualitative tool for this purpose. It is believed that NIRS can become an invaluable tool for the division of Devil's Claw into export classes, depending on HS content. This content is, however, dependant on the drying conditions used and drying parameters should be optimised to ensure the best possible quality and retention ofHS. / AFRIKAANSE OPSOMMING: Die invloed van drogingskondisies op die behoud van harpagosied, 'n iridoïed glikosied wat in die inheemse Suider-Afrikaanse plant, Harpagophytum procumbens (Duiwelsklou) voorkom, is ondersoek. Die uitvoerbaarheid van naby infra-rooi spektroskopie (NIRS) as 'n vinnige en niedestruktiewe analitiese metode vir die voorspelling van die inhoud van iridoïede, harpagosied (HS), 8-p-kumaroïel-harpagied (8pCHG) en vog, sowel as die HS:8pCHG verhouding in Duiwelsklou is ook ondersoek. Die beweerde medisinale aktiwiteit van die knolagtige sekondêre wortels van Duiwelsklou het gelei tot 'n gevestigde uitvoermark in Duitsland, en is dus van kommersiële belang. Die stabiliteit van HS gedurende prosessering is nog nooit ondersoek nie. Die huidige studie het die invloed van drie drogingsmetodes (son-, tonnel- en vriesdroging) en drie tonneldrogingstemperature (40°C, 50°C en 60°C, droëbol temperatuur) op die HS inhoud evalueer. Metanolies ekstrakte (70% metanol-water) is met hoë-druk vloeistof chromatografie ("HPLC") geanaliseer en 'n veelvoudige ekstraksie toets (n =12) het 'n 91.70% herwinning van HS getoon. Betekenisvolle (P < 0.05) verskille is gevind tussen tonnel- en vriesdroging enersyds en sondroging andersyds, met laasgenoemde die nadeligste metode. Die vergelyking van tonneldrogingstemperature, by 'n konstante relatiewe humiditeit (RH) van 30%, het getoon dat droging by 40°C die minste HS behou het en betekenisvol (P < 0.05) verskil het van droging by 50°C. Alhoewel die droging by 60°C nie betekenisvol verskil het van die ander twee kondisies nie, het dit laer HS waardes as droging by 50°C getoon. Vriesdroging het uitstekende resultate ten opsigte van die behoud van HS gelewer (1.565% ± 0.394%; gemiddelde ± standaard afwyking), maar ekonomiese beperkings plaas noodgedwonge limiete op die toepaslikheid van hierdie drogingsmetode. Verdere studie om tonneldrogingskondisies te optimiseer word is nodig (1.526% ± 0.396%), maar veral droging by 50°C en 30% RH (1.750% ± 0.438%) het prakties aanvaarbare resultate gelewer. Toekomstige navorsing moet daarop gemik word om die spesifieke degradasie meganismes van HS gedurende die droging, vervoer en opberging van Duiwelsklouwortel te verklaar. Die NIRS ontleding van gedroogde, gemaalde Duiwelsklou wortel is vergelyk met HPLC analises van dieselfde metanolise ekstrakte wat tydens die drogingstudies gebruik is. Twee verskillende NIRS instrumente is gebruik om spektra deur diffuse weerkaatsing in die poeier te meet en kalibrasies is daaropvolgend met gedeeltelike minste vierkante ("PLS") regressie gedoen. Beide onafhanklike data stel validasie en volle kruisvalidasie is gebruik om die prestasie en voorspellingsvermoë van die onderskeie modelle te evalueer. Uitstekende voginhoud (VI) kalibrasies met 'n standaard voorspellings fout (SVF) van 0.24% (klasinterval: 2.44% tot 10.43%) en 'n korrelasie van 0.99 is gevind. Die model het goed vergelyk met die standaard laboratorium fout (SLF) van 0.14%, oor die gegewe klasinterval. Met inagneming van die sensitiwiteit van HDVC as verwysingsmetode (SLF = 0.035%), het die HS inhoud kalibrasie (klasinterval: 0.693% tot 2.244%) aanvaarbaar gepresteer met 'n SVF van 0.134%. Die beskikbare klasinterval was egter relatief klein en toekomstige NIRS studies moet gebruik maak van 'n wyer klasinterval indien moontlik. Die 8pCHG model (klasinterval: 0.069% tot 0.290%) het soortgelyk gepresteer, met 'n standaard kruisvalidasie fout van 0.028% en SLF van 0.007%. Met die insluiting van monsters van 'n tweede spesie, H. zeyheri, is beide die 8pCHG inhoud en HS:8pCHG verhouding (klasinterval: 1.84 tot 34.48) kalibrasies as moontlike metodes vir onderskeiding tussen die spesies geëvalueer. Met'n 8pCHG inhoud van ca. 0.9% het die eerste model 'n goeie skeidingsvermoë getoon. In teenstelling met vorige studies egter, het die verhoudingsmodel minder sukses getoon. Hoofkomponent analise ("PCA") van die spektra was egter baie belowend as 'n kwalitatiewe onderskeidingsmetode. Volgens hierdie studie kan NIRS 'n waardevolle tegniek wees vir die verdeling van Duiwelsklou in uitvoer klasse op grond van HS inhoud. Hierdie inhoud is egter grotendeels afhanklik van die drogingskondisies en drogingsparameters wat verder verfyn behoort te word om 'n produk van die hoogste kwaliteit met behoud van HS te verseker.

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: The sensory characteristics and chemical composition of rooibos infusions were analysed to establish the extent of variation in sensory attributes and compositional parameters, and investigate whether correlations exist between the sensory characteristics and the phenolic composition. To capture as much potential variation as possible, 69 rooibos samples were collected throughout the 2009 harvesting season from different production areas. The samples were also representative of the different quality grades (A, B, C or D) which are based on the appearance of the leaves, and the colour and flavour of the infusion. Quantitative descriptive analysis was used to develop sensory profiles for each of the rooibos infusions. Considerable variation in the sensory characteristics was observed, highlighting the need for standardised terminology that can be used to describe what is often referred to as “characteristic” rooibos flavour. Aroma, flavour, taste, and mouthfeel descriptors were generated and assembled into a rooibos sensory wheel. The most recurring attributes were selected to compile a rooibos sensory lexicon which provides a definition and a reference standard for clarification of each term. A combination of “honey”, “woody” and “fynbos-floral” notes with a slightly sweet taste and subtle astringency may be regarded as “characteristic” rooibos attributes indicative of good quality tea. Pleasant “caramel” and “fruity-sweet” notes were also observed in a number of infusions, whereas negative descriptors, associated with poor quality rooibos, included “green” and “hay” flavours. Large variation in the composition of rooibos infusions was revealed through the quantification of soluble solids (SS), total polyphenols (TP), tannins and 14 monomeric phenolic compounds, as well as spectrophotometric colour measurements. High quality rooibos was associated with higher levels of SS, TP, tannins and phenolic compounds than low quality rooibos. Correlations between the compositional parameters and sensory attributes of the infusions indicated that several non-volatile compounds, including enolphenylpyruvic acid-2-glucoside, quercetin-3-glucoside, and iso-orientin, were associated with the characteristic sweet taste of rooibos, while bitterness was related to certain flavonoids such as luteolin, quercetin and aspalathin. The only compound significantly correlated to astringency was rutin, although it is likely that aspalathin and several other monomeric flavonoids also contribute to astringency. The tannin content was not associated with astringency possibly because of the limitations associated with the tannin quantification method. To determine whether compositional changes resulting from steam pasteurisation of rooibos leaves influence the sensory quality of rooibos infusions, differences in the phenolic composition and sensory attributes of infusions, prepared from unpasteurised and pasteurised rooibos, were analysed. Steam pasteurisation significantly reduced the SS, TP and aspalathin content of rooibos infusions, as well as the absorbance, especially at a wavelength of 450 nm. It also resulted in significant reductions in the intensities of most of the aroma and flavour attributes, especially the “green” notes associated with low quality rooibos. After steam pasteurisation the prominent “green” flavour of certain samples was frequently replaced by a “hay” flavour. The taste attributes, sweetness and bitterness, remained unchanged, whereas the astringency of rooibos infusions decreased significantly. / AFRIKAANSE OPSOMMING: Die sensoriese eienskappe en chemiese samestelling van rooibostee is geanaliseer ten einde die mate van variasie van die sensoriese sowel as die chemiese profiel te bepaal, asook om vas te stel of korrelasies bestaan tussen die sensoriese eienskappe en die fenoliese samestelling van rooibostee. Om soveel as moontlik potensiële variasie in te sluit, is 69 rooibos monsters tydens die 2009 oesseisoen van verskillende produksiegebiede versamel. Die monsters is ook verteenwoordigend van die verskillende kwaliteitgrade (A, B, C en D), wat op grond van die voorkoms van die teeblare, en die kleur en geur van die tee toegeken is. Kwantitatiewe beskrywende analise is gebruik om ‘n sensoriese profiel vir elke rooibostee op te stel. Aansienlike variasie in die sensoriese eienskappe is waargeneem. Dit het die behoefte aan gestandaardiseerde terminologie onderskryf, wat gebruik kan word om die term “karakteristieke” rooibos geur in meer detail te omskryf. Spesifieke terme vir aroma, geur, smaak en mondgevoel is geformuleer, en gebruik om ‘n sensoriese wiel vir rooibostee saam te stel. Terme wat die meeste voorgekom het, is gekies om in ‘n sensoriese leksikon vervat te word wat ‘n definisie en ‘n verwysingstandaard vir elke term voorstel. Die “karakteristieke” sensoriese eienskappe van goeie kwaliteit rooibostee kan soos volg gedefinieer word: ’n kombinasie van “heuning”, “houtagtige” en “fynbos-blom” geure met ‘n effense soet smaak en ‘n sagte vrankheid. Aangename “karamel” en “vrugtige-soet” geure is ook waargeneem, terwyl terme soos “groen” en “hooi” met swak kwaliteit rooibostee geassosieer is. Groot variasie in die samestelling van rooibostee is verkry deur kwantifisering van die inhoud van oplosbare vastestowwe (SS), totale polifenole (TP), tanniene, en 14 monomeriese fenoliese verbindings, asook spektrofotometriese kleurmetings. Hoë kwaliteit rooibostee is geassosieer met hoër vlakke van SS, TP, tanniene en fenoliese verbindings in vergelyking met lae kwaliteit rooibos. Korrelasies tussen laasgenoemde parameters en die sensoriese eienskappe het aangedui dat sekere nie-vlugtige verbindings, bv. fenielpirodruiwesuurglukosied, kwersetien-3-glukosied en iso-orientien, geassosieer is met die kenmerkende soet smaak van rooibos, terwyl ‘n bitter smaak geassosieer is met spesifieke flavonoïede soos luteolien, kwersetien and aspalatien. Die enigste verbinding wat betekenisvol gekorreleer het met vrankheid was rutien, maar dit is waarskynlik dat aspalatien en ander monomeriese flavonoïede ook bygedra het tot vrankheid. Die tannieninhoud het nie verband gehou met vrankheid nie, vermoedelik as gevolg van die beperkinge van die chemiese metode vir tannienbepaling. Om te bepaal of stoompasteurisasie van rooibos die sensoriese kwaliteit en chemiese samestelling van rooibostee beïnvloed, is die verskille in fenoliese samestelling en sensoriese eienskappe van die tee, berei van ongepasteuriseerde en gepasteuriseerde rooibos, ondersoek. Stoompasteurisasie het ‘n betekenisvolle verlaging teweegebring van die SS, TP en aspalatien inhoud, sowel as van die absorbansie van die tee, veral by ‘n golflengte van 450 nm. Dit het ook gelei tot ‘n statisties betekenisvolle vermindering van die intensiteit van die meeste aroma en geur eienskappe, veral die sogenaamde “groen” eienskap wat met swak kwaliteit rooibos verbind word. As gevolg van stoompasteurisasie is die opmerklike “groen” geur van sekere ongepasteuriseerde monsters herhaaldelik vervang deur ‘n “hooi” geur. Die smaakeienskappe, soet en bitter, was onveranderd, terwyl vrankheid statisties betekenisvol afgeneem het as gevolg van pasteurisasie.

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / ENGLISH ABSTRACT: Nectar formulations containing red-fleshed plum pulp and varying amounts of red-fleshed plum skin extract were developed. Red-fleshed plum nectar formulations containing 0, 8, 16, 24, and 32% skin extract were benchmarked against twenty-two commercial beverages containing red, violet and blue fruits. The total soluble solid content, pH, titratable acidity, colour, total polyphenolic, individual polyphenolic, total anthocyanin, and ascorbic acid contents, as well as antioxidant activity (oxygen radical antioxidant capacity (ORAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and ferric reducing/antioxidant power (FRAP)) of the commercial beverages and plum formulations were determined. The plum nectar formulations had similar or higher total polyphenolic content, antioxidant activity, and colour values than the average for the commercial beverages. The individual polyphenolic compounds analysed in the nectar formulations were cyanidin-3-glucoside, cyanidin-3- rutinoside, quercetin-3-glucoside, quercetin-3-rutinoside, quercetin-3-xyloside, and neochlorogenic acid. Increasing polyphenolic content and antioxidant activity was observed with an increase in skin extract content of the formulations. The sensory attributes of the formulations were plum and plantlike aroma, plum and plantlike flavour, sweetness, acidity, and astringency. Increases in plantlike aroma and flavour, acidity, and astringency in conjunction with decreases in plum aroma, plum flavour, and sweetness extract were observed with an increase in skin extract. Consumer analysis indicated that all formulations were acceptable. The 0 and 16% skin extract formulations were most preferred, while the 32% skin extract formulation was least preferred. A shelf-life study was conducted in two phases. In Phase 1, the 0, 16 and 24% skin extract formulations were stored at 0 and 5°C (analysis time points: before pasteurisation, after pasteurisation (week 0), and after 1, 2, 4, 6, 8, 12, 16, 18, 20, and 24 weeks of storage). Chemical analyses conducted included colour, total polyphenolic, individual polyphenolic compound, and total anthocyanin contents, and antioxidant activity (DPPH• scavenging activity). Results from Phase 1 indicated close associations between the 16 and 24% skin extract formulations, and between these formulations and all chemical attributes. Regression analysis of results indicated significant (P 0.05) decreases in red colour, total anthocyanins, cyanidin-3-glucoside, cyanidin-3- rutinoside, DPPH• scavenging activity, total polyphenolic content, quercetin-3-rutinoside, and quercetin- 3-xyloside for formulations stored at 0°C. The total and red colour, total anthocyanins, cyanidin-3- glucoside, and cyanidin-3-rutinoside in formulations stored at 5°C showed similar results. During Phase 2 of the shelf life study, sensory analysis was conducted on the 0 and 24% skin extract formulations stored at 5°C (preparation time points: 0, 1, 2, and 3 months). Sensory attributes, including plum, plantlike, and raisin flavour, sweetness, acidity, and astringency, were stable during storage. Chemical results from Phase 2 were similar to those of Phase 1. The chemical and sensory stability of formulations after 24 weeks of storage in Phase 1 and Phase 2 indicated that, with the exception of the anthocyanin degradation, the formulations could be beneficial to juice industries. Thus, red-fleshed plum nectars have the potential to compete with high-antioxidant fruit beverages. / AFRIKAANSE OPSOMMING: Nektarformulasies wat rooivleis pruimpulp en varieërende hoeveelhede rooivleis pruimskilekstrak bevat, is ontwikkel en ondersoek. Rooivleis pruimnektarformulasies wat 0, 8, 16, 24, en 32% skilekstrak bevat, is vergelyk met twee-en-twintig kommersiële drankies wat rooi, violet en blou vrugte bevat. Die totale oplosbare vastestof, totale polifenoliese, individuele polifenoliese, totale antosianien- en askorbiensuur inhoude, sowel as die pH, titreerbare suurheid, kleur, antioksidant aktiwiteit (suurstofradikale antioksidantkapasiteit (ORAC), 2,2-difeniel-1-pikrielhidrasiel (DPPH) radikaal blussingsaktiwiteit, en ysterreduserende/antikoksidantkrag (FRAP)) van die kommersiële drankies en pruimnektarformulasies, is bepaal. Die pruimnektarformulasies het soortgelyke of hoër totale polifenoliese inhoud, antioksidantaktiwiteit, en kleurwaardes gehad in vergelyking met die gemiddelde vir die kommersiële drankies. Die individuele polifenoliese verbindings wat in die nektarformulasies geanaliseer is, was sianidien-3-glukosied, sianidien-3-rutinosied, kwersetien-3-glukosied, kwersetien-3-rutinosied, kwersetien-3-xylosied, en neochlorogeniese suur. ‘n Toenames in die kleurwaardes, polifenoliese inhoud, en antioksidantaktiwiteit is waargeneem met ‘n toename in skilekstrak is in die formulasies. Die sensoriese eienskappe van die formulasies was pruim- en plantagtige aroma, pruim- en plantagtige geur, soetheid, suurheid, en frankheid. Toenames in plantagtige aroma en geur, suurheid en frankheid, sowel as ‘n afname in pruimaroma, pruimgeur, en -soetheid, is met ‘n toename in skilekstrak waargeneem. Verbruikersanalise het aangedui dat al die formulasies aanvaarbaar was. Die 0 en 16% skilekstrakformulasies was die mees aanvaarbaarste, terwyl die 32% skilekstrakformulasie die minste aanvaarbaar geag is. ‘n Rakleeftydstudie is in twee fases gedoen. In Fase 1 is die 0, 16, en 24% skilekstrakformulasies by 0 en 5°C gestoor (analiseringstydpunte: voor pasturisasie, na pasturisasie (week 0), en na 1, 2, 4, 6, 8, 12, 16, 18, 20, en 24 weke van berging). Chemiese analise wat gedoen is, sluit totale polifenoliese, individuele polifenoliese verbinding, en totale antosianien inhoude, sowel as kleur en antioksidantaktiwiteit (DPPH• blussingsaktiwiteit) in. Resultate van Fase 1 het ‘n nou verband tussen die 16 en 24% skilekstrakformulasies, sowel as tussen hierdie formulasies en hul chemiese kenmerke aangedui. Regressie-analise van resultate het betekenisvolle (P 0.05) afnames geïllustreer in rooi kleurwaardes, DPPH• blussingsaktiwiteit, sowel as totale antosianiene, sianidien-3-glukosied, sianidien-3-rutinosied, totale polifenoliese, kwersetien-3-rutinosied, en kwersetien-3-xylosied inhoude van die formulasies wat by 0°C gestoor is. Die totale kleur-, rooi kleurwaardes, sowel as totale antosianien, sianidien-3-glukosied, en sianidien-3-rutinosied in die formulasies wat by 5°C gestoor is, het soortgelyke resultate gegee. Gedurend Fase 2 van die rakleeftydstudie is sensoriese analise op die 0 en 24% skilekstrakformulasies wat by 5°C gestoor is, gedoen (voorbereidingstydpunte: 0, 1, 2, en 3 maande). Sensoriese eienskappe, insluitend pruim-, plantagtige-, en rosyntjiesmake, soetheid, suurheid, en frankheid, was stabiel gedurende berging. Chemiese resultate van Fase 2 was soortgelyk aan dié van Fase 1. Die chemiese en sensoriese stabiliteit van die formulasies na 24 weke van opberging in Fase 1 en Fase 2 nieteenstaande antosianienafname, het aangedui dat die formulasies voordelig kan wees vir die vrugtedrankiebedryf. Dus het rooivleis pruimnektars die vermoeë om met hoë antioksidant vrugtedrankies mee te ding.

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / Please refer to full text to view abstract.

Thesis (MSc)--Stellenbosch University, 2012. / ENGLISH ABSTRACT: The objective of this study was to investigate the impact of season on the composition and quality of male and female blesbok (Damaliscus pygargus phillipsi) muscles (Longissimus dorsi, Biceps femoris, Semimembranosus, Semitendinosus, Infraspinatus and Supraspinatus). The latter was quantified on the chemical composition (moisture, protein, fat and ash contents), fatty acid profile, mineral composition and physical attributes (pH, colour, drip and cooking loss, water holding capacity and tenderness) of the selected muscles. All of the blesbok muscles had higher (p<0.01) mean protein contents (20.6 g.100 g-1 to 23.1 g.100 g-1) when the plane of nutrition was believed to be higher (spring of 2009). A strong negative correlation (r = – 0.82; p<0.01) existed between the moisture and protein contents of the blesbok muscles. The Longissimus dorsi muscle had the highest (p<0.01) mean intramuscular fat content (3.4 g.100 g-1) when the plane of nutrition was higher. The chemical composition of the Longissimus dorsi, Biceps femoris, Semitendinosus and forequarter muscles (Infraspinatus and Supraspinatus) was affected least by the seasonal differences in the plane of nutrition and activity levels of the blesbok at the study area. However, season had a larger impact on the chemical composition of the Semimembranosus muscle. Season did not have a significant impact on the fatty acid profile of blesbok muscles, but the difference in the fatty acid profiles between male and female muscles was significant. A Principal Component Analysis (PCA) bi-plot indicated that female blesbok muscles were associated with a higher saturated fatty acid (SFA) and mono-unsaturated fatty acid (MUFA) content. Male blesbok muscles had higher (p<0.01) proportions of total polyunsaturated fatty acids (PUFA) (40.15 ± 5.39) and polyunsaturated to saturated fatty acid ratios (P:S) (0.85 ± 0.18), in comparison to female muscles (27.18 ± 8.04 and 0.54 ± 0.20, respectively). Differences in the anatomical locations of the selected blesbok muscles furthermore influenced the fatty acid profiles. The less active Longissimus dorsi muscle had higher (p<0.05) total PUFA (38.34 ± 8.62), total omega-6 (ω6) PUFA (34.46 ± 7.83), total ω3 PUFA (3.44 ± 0.84) and P:S (0.85 ± 0.24) contents, in comparison to the Infraspinatus muscle (28.96 ± 8.65, 26.23 ± 7.86, 2.31 ± 0.70 and 0.56 ± 0.19, respectively) and Supraspinatus muscle (28.85 ± 9.23, 26.05 ± 8.24, 2.28 ± 0.76 and 0.55 ± 0.21, respectively). The hindquarter muscles (Biceps femoris, Semimembranosus and Semitendinosus) had intermediate fatty acid content. Season had an impact on the calcium and zinc contents of blesbok muscles. The calcium content was higher (p<0.05) in the muscles of the animals harvested in spring (6.92 ± 1.94) compared to winter (5.61 ± 1.79). The zinc content was higher (p<0.05) in the muscles of male blesbok harvested in winter (4.04 ± 1.70) compared to spring (3.41 ± 1.67). The mineral composition was furthermore significantly different between the selected blesbok muscles. The Biceps femoris muscle had the highest (p<0.05) potassium (183.25 ± 12.79), phosphorus (180.21 ± 10.36) and magnesium (32.18 ± 1.72) content, while the sodium and calcium content was highest in the forequarter muscles. The Longissimus dorsi muscle had the highest (p<0.05) iron (3.67 ± 0.51), but significantly lower zinc content (1.63 ± 0.28), in comparison to the forequarter muscles. The pH value ≈24 h post mortem was higher (p<0.05) in the Longissimus dorsi muscle of the animals harvested in spring (5.60) compared to winter (5.54). The CIE a* (14.63 ± 0.86) and chroma (17.09 ± 0.63) values were higher (p<0.05) for winter than for male blesbok meat in spring (13.62 ± 1.08 and 16.10 ± 1.03, respectively). The latter values were also higher (p<0.05) for male compared to female (13.49 ± 0.88 and 16.22 ± 0.98) blesbok meat, at the end of the mating season (winter). The forequarter muscles had higher chroma values in comparison with the hindquarter muscles, which had higher (p<0.01) hue-angle values. Season had no influence (p<0.05) on the drip loss percentages and tenderness of blesbok muscles. The drip loss percentages were lowest (p<0.05) in the Biceps femoris and Semimembranosus muscles. The Infraspinatus and Supraspinatus muscles had the lowest (p<0.01) Warner Bratzler shear force values (20.89 ± 3.23 and 24.90 ± 5.35 N, respectively). Seasonal differences in the chemical composition of blesbok muscles were statistically significant. However, these differences were numerically small and it is therefore debatable whether they are of any biological relevance relating to human nutrition. The differences in the fatty acid profile and mineral composition as well as the physical meat quality attributes of blesbok muscles were more attributed to differences in the anatomical locations of the selected muscles, as opposed to the impact of season or gender. / AFRIKAANSE OPSOMMING: Die doel van die studie was om die impak van seisoen op die samestelling en kwaliteit van blesbok (Damaliscus pygargus phillipsi) spiere (Longissimus dorsi, Biceps femoris, Semimembranosus, Semitendinosus, Infraspinatus en Supraspinatus) te bepaal. Die seisoenale impak was gekwantifiseer op die chemiese samestelling (vog-, proteïen-, vet- en asinhoud), vetsuurprofiel, mineraal samestelling en fisiese eienskappe (pH, kleur, drup- en kookverlies, water houvermoë en taaiheid) van die geselekteerde spiere. Met ʼn hoër voedingspeil (lente 2009) het elkeen van die spiere gemiddeld ʼn hoër (p<0.01) proteïeninhoud (20.6 g.100 g-1 tot 23.1 g.100 g-1) gehad. ‘n Sterk negatiewe korrelasie (r = – 0.82; p<0.01) het bestaan tussen die vog- en proteïeninhoud van die blesbokspiere. Met ʼn hoër voedingspeil het die Longissimus dorsi spier die hoogste (p<0.01) gemiddelde intramuskulêre vetinhoud (3.4 g.100 g-1) gehad. Seisoenale verskille in die voedingspeil en aktiwiteitsvlakke van dié blesbokke het minimale verskille in die chemiese samestelling van die Longissimus dorsi, Biceps femoris, Semitendinosus en voorkwartspiere (Infraspinatus en Supraspinatus) tot gevolg gehad. Daar was wel seisoenale verskille in die chemiese samestelling van die Semimembranosus spier. Seisoen het nie ʼn beduidende invloed op die vetsuurprofiel van die blesbokspiere gehad nie, maar daar was wel beduidende (p<0.05) verskille tussen geslagte. Soos aangedui deur ʼn hoofkomponent-analise (PCA) bi-plot, was die spiere van die vroulike blesbokke meer geassosieer met hoër versadigde en mono-onversadigde vetsuursamestellings. Die spiere van die manlike diere het hoër (p<0.01) proporsies poli-onversadigde vetsure (PUFA) (40.15 ± 5.39) asook hoër poli-onversadigde tot versadigde vetsuur verhoudings (P:S) gehad (0.85 ± 0.18) in vergelyking met die spiere van die vroulike diere (onderskeidelik 27.18 ± 8.04 en 0.54 ± 0.20). Die vetsuurprofiel van blesbokspiere was ook beïnvloed deur die anatomiese ligging van die spiere. Die minder aktiewe Longissimus dorsi spier het ʼn hoër (p<0.05) totale PUFA (38.34 ± 8.62), totale omega-6 (ω6) PUFA (34.46 ± 7.83), totale ω3 PUFA (3.44 ± 0.84) en P:S (0.85 ± 0.24) inhoud gehad in vergelyking met die Infraspinatus spier (onderskeidelik 28.96 ± 8.65, 26.23 ± 7.86, 2.31 ± 0.70 en 0.56 ± 0.19) en Supraspinatus spier (onderskeidelik 28.85 ± 9.23, 26.05 ± 8.24, 2.28 ± 0.76 en 0.55 ± 0.21). Die agterkwartspiere (Biceps femoris, Semimembranosus en Semitendinosus) het intermediêre vetsuursamestellings gehad. Seisoen het ʼn invloed op die kalsium- en sinkinhoud van die blesbokspiere gehad. In die lente het die spiere gemiddeld ʼn hoër (p<0.05) kalsiuminhoud gehad (6.92 ± 1.94), in vergelyking met dié van winter (5.61 ± 1.79). Die manlike spiere van die blesbokke wat in winter geoes is, het weer ʼn hoër (p<0.05) sinkinhoud (4.04 ± 1.70) in vergelyking met dié van die lente (3.41 ± 1.67) gehad. Verder het die mineraalinhoud van die geselekteerde blesbokspiere betekenisvol van mekaar verskil. Die Biceps femoris spier het die hoogste (p<0.05) kalium- (183.25 ± 12.79), fosfor- (180.21 ± 10.36) en magnesiuminhoud (32.18 ± 1.72) gehad. Die natrium- en kalsiuminhoud was hoër in die voorkwartspiere. Die Longissimus dorsi spier het die hoogste (p<0.05) ysterinhoud (3.67 ± 0.51) gehad. Laasgenoemde het ʼn beduidend laer sinkinhoud (1.63 ± 0.28) in vergelyking met die voorkwartspiere gehad. Die Longissimus dorsi spiere van die blesbokke wat in die lente geoes is, het gemiddeld hoër pH-waardes by ≈24 uur post mortem gehad (5.60) in vergelyking met die pH-waardes van dié spiere in winter (5.54). Die CIE a*- (14.63 ± 0.86) en chroma-waardes (17.09 ± 0.63) van die manlike blesbokspiere was hoër (p<0.05) in die winter as in die lente (onderskeidelik 13.62 ± 1.08 en 16.10 ± 1.03). Aan die einde van die paartyd (winter) het die manlike blesbokke se spiere ook hoër (p<0.05) CIE a*- en chroma-waardes as die vroulike blesbokspiere (13.49 ± 0.88 en 16.22 ± 0.98) gehad. Die voorkwartspiere het gemiddeld hoër (p<0.05) chroma-waardes as die agterkwartspiere gehad, maar laasgenoemde het weer hoër (p<0.01) hue-angle waardes as die voorkwartspiere gehad. Seisoen het geen effek (p<0.05) op die drupverlies persentasies en taaiheid van die blesbokspiere gehad nie. Die Biceps femoris en Semimembranosus spiere het wel die laagste (p<0.05) drupverlies persentasies gehad. Die Infraspinatus en Supraspinatus spiere het weer die laagste (p<0.01) taaiheid (onderskeidelik 20.89 ± 3.23 en 24.90 ± 5.35) in vergelyking met die Longissimus dorsi, Biceps femoris, Semimembranosus en Semitendinosus spiere gehad (onderskeidelik 30.57 ± 6.69, 27.35 ± 3.42, 28.65 ± 4.48 en 31.51 ± 5.63). Alhoewel daar in die studie statisties beduidende seisoenale verskille in die chemiese samestelling van die blesbokspiere was, is die verskille numeries klein en is dit debatteerbaar of dié verskille enigsins biologies van toepassing is op menslike voeding. Verder het die anatomiese ligging van die geselekteerde blesbokspiere in die studie ʼn groter invloed op die verskille in die vetsuurprofiel, mineraal samestelling asook die fisiese eienskappe van die spiere gehad, in vergelyking met die impak van die oes-seisoen en die effek van geslag.

Thesis (MSc Food Sc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: Over the past two decades, there has been an increase in the use of water sources for irrigation, as well as an increase in Escherichia coli outbreaks linked to fresh produce. The full extent and type of E. coli contamination present in natural water sources is unknown and the contamination sources have also not been confirmed. The aim of this study was to enumerate and characterise E. coli from both irrigation water and potential contamination source sites. Total coliform and E. coli counts found in contamination source sites were as high as log 7.114 and log 6.912 MPN.100 mL-1, respectively. Total coliform and E. coli counts for irrigation sites were lower, with maximum counts of log 5.788 and log 5.768 MPN.100 mL-1, respectively. It was found that more than one third (5/14 = 35.71%) of the irrigation sites had E. coli counts exceeding the guidelines (<1 000 counts.100 mL-1) for ‘safe’ irrigation water for fresh produce (<1 000 counts.100 mL-1) as set by the Department of Water Affairs (DWA) and World Health Organisation (WHO), making the water unsuitable for the irrigation of fresh produce. Phylogenetic subgroups (A0, A1, B1, B22, B23, D1 and D2) and the MALDI Biotyper system (PCA dendrogram) were used to create a fingerprint of each E. coli isolated from the environment. These were then used to link E. coli strains from irrigation water to their most probable contamination origin. Escherichia coli population structure was found in this study, to be better suited for linking E. coli strains from irrigation water to their most likely source, than just applying the phylogenetic grouping. The MALDI Biotyper data in combination with the phylogenetic subgroup assignment was then used to group similar strains and link E. coli from irrigation water to their contamination sources by comparing population structures. Strains isolated from surface and groundwater showed similar distribution patterns, but groundwater strains showed a population structure more indicative of porcine and bovine origin, while surface water showed population characteristics which could not be used to make conclusive links between the irrigation water and suspected contamination sources. When investigating the population structures of individual sample sites, it was found that phylogenetic subgroups A0, A1 and B1 frequently made up the bulk of the E. coli population. It was also found that linking individual irrigation sites to contamination sources was successful, as irrigation site Berg-2 was found to have a similar population structure to contamination source site Plank-1 which represents human pollution from an informal settlement. This led to the conclusion that Berg-2 was being contaminated by human pollution, most probably from an informal settlement. Upon further investigation it was found that Berg-2 is downstream of an informal settlement, proving that E. coli population structure is a successful means of microbial source tracking (MST). Virulence factors of the 153 E. coli isolated during the study were identified and the potential risk associated with using the investigated irrigation water for irrigation of fresh produce, was determined. Two enteropathogenic E. coli (EPEC) strains were isolated from the irrigation water, one from the Plankenburg River water, and the other from a borehole in the Drakenstein area. The latter indicates that borehole water is not as safe as was once thought, and that there are bacterial contaminants finding their way into groundwater. The occurrence of an EPEC strain in river water shows that neither ground nor surface water is guaranteed to be safe, and that treatment of water being used for the irrigation of fresh produce should be implemented. / AFRIKAANSE OPSOMMING: Oor die afgelope twee dekades was daar nie net 'n toename in die gebruik van alternatiewe waterbronne vir besproeiing nie, maar ook 'n toename in uitbrake van Escherichia coli uitbrake wat aan vars produkte gekoppel kan word. Die tipe E. coli-besmetting wat in natuurlike waterbronne teenwoordig is, is onbekend en die besmettingsbron is ook nog nie bevestig nie. Daarom was die doel van hierdie studie om die voorkomssyfer van E. coli van beide besproeiingswater en potensiële kontaminasiebronne te bepaal, asook om die E. coli te karakteriseer. Totale kolivorme en E. coli-tellings wat in kontaminasiebronne gevind is, het ‘n maksimum van log 7,114 en log 6,912 MPN.100 mL-1 onderskeidelik bereik, terwyl die totale kolivorme en E. coli-tellings vir besproeiingswater laer was, met 'n maksimum van log 5,788 en 5,768 MPN.100 mL-1, onderskeidelik. Dit is bevind dat meer as 1/3 (5/14 = 35,71%) van die besproeiingswaterbronne meer E. coli bevat as wat toegelaat word in die riglyne vir "veilige" besproeiingswater vir vars produkte (<1 000 fekale koliforme.100 mL-1) wat deur die Departement Waterwese (DWA) en die Organisasie vir Wêreldgesondheid (WHO) aanbeveel word. Filogenetiese subgroepe (A0, A1, B1, B22, B23, D1 en D2) en die ‘MALDI Biotyper’-stelsel (PKA dendrogram) is gebruik om unieke profiele vir elke geïsoleerde E. coli te skep. Dié profiele is daarna gebruik om E. coli-stamme van besproeiingswater te koppel aan die mees waarskynlike oorsprong van kontaminasie. Daar is in hierdie studie bevind dat die E. coli-populasiestruktuur beter geskik was vir die koppeling van E. coli-stamme van besproeiingswater na hul mees waarskynlikste bron, as net die toepassing van die filogenetiese groepering. Dit was toe gevind dat E. coli wat uit oppervlak- en grondwater geïsoleer is, soortgelyke verspreidingspatrone het, maar grondwaterstamme se bevolkingstruktuur is meer aanduidend van fekale besmetting deur varke en beeste, terwyl oppervlakwater se bevolkingseienskappe nie duidelik genoeg was om ‘n gevolgtrekking oor moontlike bronne van besmetting te maak nie. Toe die populasiestruktuur van die individuele bemonsteringspunte ondersoek is, is daar bevind dat die filogenetiese subgroepe A0, A1 en B1 dikwels die meeste tot die E. coli bevolking bydra. Daar is ook bevind dat die koppeling van isolate in individuele besproeiingswaterbronne met hul mees waarskynlike bronne van kontaminasie suksesvol was. Besproeiingswater van Berg-2 het 'n soortgelyke populasiestruktuur as Plank-1 wat beskou is as ‘n kontaminasiebron. Dit het gelei tot die gevolgtrekking dat Berg-2 heel waarskynlik deur menslike besoedeling beïnvloed word, soos Plank-1, en dat daar moontlik ook ‘n informele nedersetting by Berg-2 betrokke is. Na verdere ondersoek is gevind dat Berg-2 inderdaad ook stroomaf van 'n ander informele nedersetting geleë is, wat bewys dat die E. coli-populasiestruktuur 'n suksesvolle manier is om E. coli kontaminasie te verbind met besproeiingswater. Patogeniese faktore, wat in E. coli voorkom en maagkieme veroorsaak, is vooraf getoets in elkeen van die 153 E. coli-isolate wat tydens die studie geïdentifiseer is. Twee ‘enteropathogenic’ E. coli (EPEC)-stamme is uit die besproeiingswater geïsoleer: een uit die Plankenburgrivier (Plank-3), en die ander uit 'n boorgat in die Drakenstein-gebied (Boorgat A1). Hierdie inligting dui aan dat boorgatwater nie so veilig is as wat voorheen vermoed is nie, en dat bakteriese kontaminasie wel vookom wat nie alleen die grondwater besmet nie, maar ook daarin oorleef. Die voorkoms van die EPEC-stamme in hierdie studie is ‘n aanduiding dat beide grond- en opppervlakwater ewe gevaarlik kan wees, en dat daar dus geen waarborg vir die veiligheid van die water is nie. Die behandeling van grond- en oppervlakwater, wat vir die besproeiing van vars produkte gebruik word, moet daarom ernstig oorweeg word om moontlike uitbrake van E. coli op vars produkte te verhoed.

Thesis (MSc Food Sc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: A wide variety of traditionally and commercially fermented milks are commonly consumed in various countries of Sub-Saharan Africa. Commercially fermented milk is produced on an industrial scale according to well-managed, standardised production processes and starters are used to initiate fermentation. Traditionally fermented milk is prepared domestically and fermentation occurs spontaneously at ambient temperatures. Lactic acid bacteria (LAB) are responsible for milk fermentation during which they convert the milk carbohydrates to lactic acid, carbon dioxide, alcohol and other organic metabolites. Acetic acid bacteria (AAB), yeasts and mycelial fungi have also been isolated from fermented milks. In this study the microbial consortium present in three traditionally fermented milks, namely omashikwa from Namibia, masse from Mozambique and chekapmkaika from Uganda and two commercially fermented milks, namely chambiko from Malawi and omaere from Namibia, were isolated and enumerated on six different selective media that included MSR + C (specific for lactobacilli), KCA + TTC (specific for lactococci), KCA + V (specific for leuconostocs), MRS + E (specific for AAB), MEA (specific for mycelial fungi) and YPD (specific for yeasts). No significant differences were found between the enumeration values obtained for the three chambiko samples, as well as for enumeration values obtained for the two omaere samples on each of the selective media, indicating low sample variance. Significant differences between enumeration values obtained for the three omashikwa samples were found on all six selective media. Significant differences between enumeration values of the three masse samples and both the chekapmkaika samples were also observed on the selective media. In addition to this, significant differences were observed between average enumeration values obtained for each media between the masse and chekapmkaika, the chambiko and omaere, as well as when the traditional and commercial milks were compared. According to the average enumeration values obtained on each media selective for LAB, the highest bacterial counts were detected on KCA + TTC medium for omaere (2.3 x 106 cfu.ml-1), KCA + V for chambiko (1.8 x 105 cfu.ml-1), KCA + TTC for omashikwa and MRS + C for masse and chekapmkaika (6.2 x 106 and 2.0 x 103 cfu.ml-1, respectively). After isolation and enumeration of the microbes present in each milk, bacterial isolates on the media selective for LAB and AAB were obtained according to the Harrison Disk method. These isolates were identified by amplifying a 1.5 kilobase (kb) part of the 16S ribosomal RNA (rRNA) gene using the polymerase chain reaction (PCR), followed by DNA sequencing. The isolates were identified by comparing the sequences obtained to sequences listed in the NCBI database using the BLAST algorithm and searching for the closest relative. The main LAB group present in the omaere was lactococci (94%), in chambiko and chekapmkaika it was lactobacilli (30% and 45%, respectively), in omashikwa it was enterococci (43%) and in masse it was leuconostocs (68%). The same microbial species were present on a number of the selective media used in this study. Lactococcus spp., Enterococcus spp. and Lactobacillus spp. were isolated from MRS + C, KCA + TTC, KCA + V and MRS + E and Leuconostoc spp. were isolated from MRS + C, MRS + E and KCA + V. Hygienic standards during traditional milk fermentation is often poor and, therefore, microbial contaminants were isolated from the traditional milk and these included Acinetobacter johnsonii and Klebsiella pneumoniae from KCA + V, Mesorhizobium loti, Acinetobacter radioresistens, Escherichia coli, Staphylococcus spp., Kluyvera georgiana, Enterobacter spp. and Klebsiella oxytoca from KCA + TTC, Staphylococcus spp. from MRS + C and Bacillus spp. from MRS + E. Since the media used for the isolation of the LAB and AAB in this study were not selective further identification of the enumerated microbes is of importance for the identification of the microbial groups present in each fermented milk. The data obtained in this study clearly shows that fermented milks from Sub-Saharan Africa vary significantly from each other in terms of microbial numbers, microbial diversity and the dominant microbial groups present. The microbial diversity of the traditionally fermented milks was more diverse than the microbial diversity of the commercially fermented milks. LAB strains isolated from these traditionally fermented milks can be used to develop novel starters and as a result new commercially fermented dairy products with unique aromas, tastes and characteristics can be produced. / AFRIKAANSE OPSOMMING: 'n Wye verskeidenheid tradisioneel en kommersieel gefermenteerde melk produkte word algeneem verbruik in verskeie lande van Sub-Sahara Afrika. Kommersieel gefermenteerde melk word geproduseer op groot skaal, deur deeglik bestuurde gestandardiseerde produksieprosesse en 'n beginkultuur word gebruik om fermentasie te inisieer. Tradisioneel gefermenteerde melk word tuis gemaak en fermentasie gebeur spontaan by kamertemperatuur. Melksuurbakterieë (MSB) is verantwoordelik vir melkfermentasie waartydens die bakterieë koolhidrate omskakel na melksuur, koolstofdioksied, alkohol en ander organiese sure. Asetaatsuurbakterieë (ASB), giste en miseliale fungi is ook al van gefermenteerde melk geïsoleer. In hierdie studie is die mikrobiese konsortium teenwoordig in drie soorte tradisioneel gefermenteerde melk, naamlik omashikwa van Namibië, masse van Mosambiek en chekapmkaika van Uganda en twee soorte kommersieel gefermenteerde melk, naamlik chambiko van Malawi en omaere van Namibië, geïsoleer en getel op ses verskillende selektiewe groeimedia insluitend MRS + C (spesifiek vir lactobacilli), KCA + TTC (spesifiek vir lactococci), KCA + V (spesifiek vir leuconostocs), MRS + E (spesifiek vir ASB), MEA (spesifiek vir miseliale fungi) en YPD (spesifiek vir giste). Geen betekenisvolle verskille is gevind tussen die mikrobiese tellings verkry vir die drie chambiko monsters nie, sowel as tussen die mikrobiese tellings verkry vir die twee omaere monsters, op elk van die selektiewe groeimedia, wat dui op lae monster variansie. Betekenisvolle verskille is gevind tussen die mikrobiese tellings verkry vir die drie omashikwa monsters op al ses selektiewe groeimedia. Betekenisvolle verskille is ook waargeneem tussen die mikrobiese tellings van die drie masse monsters en beide die chekapmkaika monsters op die selektiewe groeimedia. Daarbenewens is betekenisvolle verskille waargeneem tussen gemiddelde mikrobiese tellings verkry vir elke groeimedium tussen die masse en chekapmkaika, die chambiko en omaere asook toe die tradisionele en kommersiële melk produkte met mekaar vergelyk is. Volgens die gemiddelde mikrobiese tellings verkry op elk van die groeimedia selektief vir MSB, is die hoogste mikrobiese telling waargeneem op KCA + TTC medium vir omaere (2.3 x 106 kve.ml-1), KCA + V vir chambiko (1.8 x 105 kve.ml-1), KCA + TTC vir omashikwa en MRS + C vir masse en chekapmkaika (6.2 x 106 en 2.0 x 103 kve.ml-1, respektiewelik). Na die isolasie en tel van die mikrobes teenwoordig in elke melk is bakteriese isolate op die media selektief vir MSB en ASB verkry volgends die Harrison Disk metode. Hierdie isolate is geïdentifiseer deur amplifikasie van „n 1.5 kilobasis (kb) gedeelte van die 16S ribosomale RNS (rRNS) geen deur gebruik te maak van die polimerase kettingreaksie gevolg deur DNS klonering. Die isolate is geïdentifiseer deur die gekloneerde insetsels se volgordes te vergelyk met volgordes beskikbaar op die NCBI webwerf deur van die BLAST algoritme gebruik te maak en die naas verwante insetsel op te spoor. Die hoof MSB groep teenwoordig in die omaere was lactococci (94%), in chambiko en chekapmkaika was dit lactobacilli (30% en 45%, respektiewelik), in die omashikwa was dit enterococci (43%) en in die masse was dit leuconostocs (68%). Dieselfde mikrobiese spesies was teenwoordig op verskeie van die selektiewe groeimedia gebruik in hierdie studie. Lactococcus spp., Enterococcus spp. en Lactobacillus spp. is geïsoleer van MRS + C, KCA + TTC, KCA + V en MRS + E en Leuconostoc spp. is geïsoleer van MRS + C, MRS + E en KCA + V. Higiëniese standaarde tydens tradisionele melkfermentasie is dikwels swak en dus is mikrobiese kontaminante geïsoleer van die tradisionele melk produkte insluitend Acinetobacter johnsonii en Klebsiella pneumoniae van KCA + V, Mesorhizobium loti, Acinetobacter radioresistens, Escherichia coli, Staphylococcus spp., Kluyvera georgiana, Enterobacter spp. en Klebsiella oxytoca van KCA + TTC, Staphylococcus spp. van MRS + C en Bacillus spp. van MRS + E. Aangesien die media wat gebruik is vir die isolasie van die MSB en ASB in hierdie studie nie selektief was nie, is verdere identifikasie van die getelde mikrobes belangrik vir die identifikasie van die mikrobiese groepe teenwoordig in elke melk. Die data verkry in hierdie studie dui aan dat gefermenteerde melk produkte van Sub-Sahara Afrika betekenisvol van mekaar verskil in terme van mikrobiese getalle, mikrobiese diversiteit en die dominante mikrobiese groepe teenwoordig. Die mikrobiese diversiteit van die tradisioneel gefermenteerde melk produkte was meer divers as die mikrobiese diversiteit van die kommersieel gefermenteerde melk produkte. MSB spesies geïsoleer van hierdie tradisioneel gefermenteerde melk produkte kan gebruik word om nuwe beginkulture te ontwikkel en gevolglik kan nuwe kommersieel gefermenteerde suiwelprodukte met unieke aromas, smake en eienskappe geproduseer word.