Thesis (MSc)--Stellenbosch University, 2015. / ENGLISH ABSTRACT: Acid mine drainage (AMD) is wastewater generated by mine and industrial activity with typically high heavy metal and sulphur content potentially resulting in toxic wastewater upon exposure to dissolved oxygen, water and micro-organisms. Due to the hazardous consequences of untreated AMD, treatment methods such as semi-passive biotic treatments, including constructive wetlands and microbial bioreactors were developed. Microbial bioreactors rely on suitable carbon sources such as ethanol, grasses and manure and the creation of anaerobic conditions for the reduction of sulphate, chemical oxidizable organic matter (COD) and to neutralise pH. Domestic wastewater sludge has also been identified as an economical and readily available carbon source that allows the treatment of both AMD and domestic wastewater. A synthetic medium simulating the COD and the biological degradable organic matter (BOD) of domestic wastewater sludge was formulated to exclude variations in the evaluation of domestic wastewater sludge as carbon source in the treatment of AMD. Firstly the BOD and COD of anaerobic domestic wastewater sludge was determined and used as parameters in the formulation of the synthetic medium. A ratio of 1:1 AMD: synthetic domestic wastewater sludge (SDWWS) was the optimum ratio in terms of sulphate and COD removal. Secondly, medical drip bags were used as anaerobic bioreactors to determine the microbial diversity in AMD treated with SDWWS using different variables. Data analyses from next generation sequencing showed that Chlorobium spp. dominated the 90 d pioneer trials at relative percentages of 68 % and 76 %. Transmission electron microscopy (TEM) images and the bright green colour of the liquid contents confirmed the data analyses. Sulphates and COD were removed at > 98 % and > 85 %, respectively. A shorter incubation time was investigated in the 30 d pioneer trial. Chlorobium spp. was dominant, followed by Magnetospirillum spp. and Ornithobacterium spp. The liquid content changed to a dark brown colour. COD and sulphate concentrations were reduced by 60.8 % and 96 %, respectively, within 26 d, after which a plateau was reached. The effect of an established biofilm in the bioreactors showed that Chlorobium spp. also dominated approximately 62 %, in comparison to the 36 % in the 30 d pioneer trial. A sulphate and COD reduction of 96 % and 58 %, respectively, was obtained within 26 d and the liquid content was the same colour as in the 30 d pioneer trial. It is possible that brown Green sulphur bacteria were present. Therefore, although Chlorobium spp. was present at a higher percentage as in the 30 d pioneer trial, the removal of COD and sulphate was similar. During the 30 d trials a white precipitant formed at the top of the bioreactors, consisting primarily of sulphate and carbon that was also indicative of the presence of Chlorobium spp. Incubation at reduced temperature reduced sulphates by only 10 % and COD by 12 % after 17 d, followed by a plateau. Ornithobacterium spp. dominated in the first trial and Magnetospirillum spp. in the second trial. / AFRIKAANSE OPSOMMING: Suur mynwater (SMW) is afvalwater wat deur die myn- en industriële bedryf gegenereer word en bevat kenmerklik hoë konsentrasies swaar metale en swawel wat potensieel in toksiese afvalwater omskep kan word indien blootgestel aan opgelosde suurstof, water en mikro-organismes. Die skadelike gevolge wat blootstelling aan onbehandelde SMW mag hê, het gelei tot semi-passiewe behandelinge wat vleilande en mikrobiese bioreaktors insluit. Mikrobiese bioreaktore maak staat op n geskikte koolstofbron soos etanol, grasse en bemesting en die skep van ‘n anaerobiese omgewing vir die verwydering van sulfate en chemies oksideerbare organiese material (CSB), asook die neutralisering van pH. Huishoudelike afvalwaterslyk is ook uitgewys as ‘n ekonomies geskikte en algemeen beskikbare koolstofbron wat die behandeling van beide SMW en huishoudelike afvalwater toelaat. ‘n Sintetiese medium wat die CSB en biologies afbreekbare organiese materiaal (BSB) van huishoudelike afvalwater slyk naboots is geformuleer om die variasies in die evaluasie van huishoudelike afvalwater slyk as koolstofbron vir die behandeling van SMW, uit te sluit. Eerstens is die BSB en die CSB van huishoudelike afvalwater slyk bepaal en gebruik as n maatstaf vir die formulering van die sintetiese medium. ‘n Verhouding van 1:1 sintetiese huishoudelike afvalwater slyk (SDWWS) en SMW is optimaal ratio i.t.v. die verwydering van sulfate en CSB. Tweedens is mediese dripsakkies as anaerobiese bioreaktore gebruik om die mikrobiese diversiteit in SMW, wat met SDWWS behandel is, te bepaal deur verskeie veranderlikes te gebruik. Tweede generasie DNA-volgorde bepalingstegnieke is gebruik en data analises het gewys dat Chlorobium spp. die 90 d pionier toetslopie domineer met relatiewe persentasies van 68 % en 76 %. Transmissie elektron mikroskopie fotos en die helder groen kleur van die dripsakkies se vloeistof inhoud het die data analises bevestig. Die sulfate en CSB inhoud is onderskeidelik met > 98 % en > 85 % verminder. ‘n Korter behandelingstydperk is ondersoek met n 30 d pionier toetslopie. Chlorobium spp. was dominant, gevolg deur Magnetospirillum spp. en Ornithobacterium spp. Die vloeistof inhoud het na ‘n donker bruin kleur verander. Die CSB en sulfaat konsentrasies is met 60.8 % en 96 % onderskeidelik verminder na 26 dae waarna ‘n plato bereik is. Die effek van ‘n reeds bestaande biofilm in die bioreaktore het gewys dat Chlorobium spp. ook gedomineer het teen ‘n relatiewe persentasie van 62 % in vergelyking met die 36 % in die 30 d pionier toetslopie. ‘n Vermindering in sulfate en CSB van 96 % en 58 % is onderskeidelik is bereik binne 26 d en die vloeistofinhoud was dieselfde kleur as die bioreaktore in die 30 d pionier toetslopie. Dit is moontlik dat die bruin Groen swawel bakterieë teenwoordig was. Daarom, ondanks ‘n groter teenwoordigheid van die Chlorobium spp. teen ‘n relatiewe persentasie in vergelying met die 30 d pionier toetslopie, was die verwydering van CSB en sulfate soortgelyk. Tydens die 30 d toetslopies het ‘n wit neerslag aan die bokant van die bioreaktore gevorm wat hoofsaaklik uit sulfaat en koolstof bestaan het wat ook ‘n aanduiding van die teenwoordigheid van Chlorobium spp. is. ‘n Toetslopie wat by laer temperature uitgevoer is kon die sulfate en CSB met slegs 10 % en 12 % onderskeidelik verminder nadat ‘n plato na 17 d bereik is. Ornithobacterium spp. het die eerste toetslopie gedomineer waar Magnetospirillum spp. die tweede toetslopie gedomineer het.

Thesis (PhD (Food Science))--University of Stellenbosch, 2006. / High-rate anaerobic bioreactors are used for the treatment of various wastewaters, of which the upflow anaerobic sludge blanket (UASB) bioreactor has the widest application, especially in the food and beverage industries. In an UASB bioreactor sludge develops in a particular granular or flocculent form and the success of the anaerobic process relies on the formation of active and settable granules. These granules are formed by self-aggregation of bacteria that can be divided into different trophic groups that are responsible for the metabolic breakdown of organic substrates. The successful performance of a bioreactor is influenced by the composition of the substrate which subsequently may have an impact on the microbial consortium present in the UASB granules. In order to determine if a change in the structure of the non-methanogenic microbial community takes place, UASB brewery granules were subjected to the sudden addition of different carbon sources at different concentrations. A shift in the microbial community did occur when the granules were subjected to lactate medium (5 g.l-1). No changes in the microbial community were observed when the granules were stressed with glucose medium as carbon source, regardless of an increase in the glucose concentration. In order to better understand the effect that different wastewaters may have on the microbial consortium present in different UASB granules, the polymerase chain reaction (PCR) based denaturing gradient gel electrophoresis (DGGE) technique and sequence analysis were used to fingerprint and identify the Bacteria and Archaea present in either, winery, brewery, distillery or peach-lye canning UASB granules. Each granule type showed distinct PCR-based DGGE fingerprints with unique bands, while other bands were found to be present in all the granules regardless of the wastewater being treated. Bacillus, Pseudomonas, Bacteroides, Enterococcus, Alcaligenes, Clostridium, Shewanella, Microbacterium, Leuconostoc, Sulfurospirillum, Acidaminococcus, Vibrio, Aeromonas, Nitrospira, Synergistes, Rhodococcus, Rhodocyclus, Syntrophobacter and uncultured bacteria were identified, representing different acidogenic, acetogenic and homoacetogenic Bacteria.Different methanogenic bacteria such as Methanosaeta, Methanosarcina, Methanobacterium and uncultured bacteria belonging to the group Archaea were also fingerprinted and identified from different UASB granules. In both these studies a DGGE marker was constructed that may be used to assist in the identification of bacteria. The DGGE marker can also be used to monitor the presence of bacteria over a time period during anaerobic digestion. Bioaugmentation or the enrichment of granules results in tailor-made granules that may be used for the treatment of specific wastewaters. One of the most important contributions to the maintenance and enhancement of UASB granule formation is the inclusion of suitable microbes in the granule structure. Enterobacter sakazakii was isolated from raw winery wastewater and was found to produce sufficient amounts of desired fatty acids. This bacteria was, therefore, incorporated into batch cultured granular sludge. In order to identify and monitor the presence of the incorporated E. sakazakii in the tailor-made granules, 16S rRNA gene sequence primers and PCR conditions were developed. The use of molecular techniques such as PCR-based DGGE and sequence analysis proved to be successful methods to fingerprint and identify the microbial consortium present in the different UASB granules.

Οι βιοαντιδραστήρες μεμβρανών αποτελούν μια καινοτόμο τεχνολογία, ιδανική για την αποκατάσταση προηγμένων αστικών και βιομηχανικών λυμάτων, τα οποία περιέχουν υψηλά ποσοστά βιοαποικοδομήσιμων οργανικών ενώσεων. Η παρούσα εργασία αποτελεί ένα μέρος μιας ευρύτερης προσπάθειας ανάπτυξης μιας νέας κατηγορίας λειτουργικών μεμβρανών τεχνολογίας “Βιοαντιδραστήρα Μεμβρανών” (Membrane Bioreactors, MBRs) ή/και βελτίωσης των ήδη χρησιμοποιούμενων μεμβρανών τεχνολογίας MBR με την ενσωμάτωση στο ενεργό πορώδες τους νανοσωλήνες άνθρακα. Οι νανοσωλήνες άνθρακα δυνητικά θα προσέφεραν ταυτόχρονα υψηλότερες ταχύτητες ροής νερού, υψηλότερο ποσοστό απόρριψης οργανικών ενώσεων και ανόργανων αλάτων χαμηλού μοριακού βάρους, καθώς και υψηλότερη αντοχή της μεμβράνης στην εναπόθεση διαφόρων μικροοργανισμών. Η πρόκληση στην περίπτωση αυτή είναι η αποτελεσματική ενθυλάκωση τους στην ενεργή εκλεκτική στοιβάδα των μεμβρανών αυτών. Οι νανοσωλήνες άνθρακα από την πρώτη στιγμή της ανακάλυψης τους, έχουν προσελκύσει το ενδιαφέρον της επιστημονικής κοινότητας, λόγω της ευρείας εφαρμογής τους σε πολλά επιστημονικά και τεχνολογικά πεδία, ως συνέπεια των μοναδικών ιδιοτήτων τους. Οι χημικές, οπτικές, ηλεκτρικές και μηχανικές ιδιότητές τους, τους καθιστούν δυνητικά χρήσιμους σε πάρα πολλές εφαρμογές. Στη συγκεκριμένη περίπτωση, τα τελευταία 5-7 έτη, οι νανοσωλήνες άνθρακα έχουν ταυτοποιηθεί ως μια καινούργια γενιά νανο-πορωδών υλικών με τρομερό δυναμικό για εφαρμογές ως φίλτρα σε υλικά μεμβρανών που θα μπορούσε να φέρει πραγματική επανάσταση στο σχετικό χώρο. H δυνατότητα ελέγχου της διαμέτρου τους και κατά συνέπεια του μεγέθους των πόρων τους μέσω των οποίων λαμβάνει χώρα το φαινόμενο της διάχυσης ή ροής (από τα 4 Angstroms έως τα 15 nm), σε συνδυασμό με τα σχεδόν άτριβου χαρακτήρα γραφιτικά τους τοιχώματα, εξασφαλίζει εξαιρετικά ταχεία ροή μικρών μορίων με ταυτόχρονη καταπληκτική εκλεκτικότητα στη διαπερατότητα μορίων με βάση το μέγεθός τους. Η ροή υγρών μέσα από αυτές των νανοσωλήνων άνθρακα προβλέπεται να είναι 3-5 τάξεις μεγέθους πάνω απ’ ότι αναμένεται με βάση υπολογισμούς βασισμένους σε απλές αρχές της υδροδυναμικής. Στο πλαίσιο αυτό, μελετήθηκε ο εμβολισμός νανοπορωδών εμπορικών μεμβρανών με διάφορα είδη νανοσωλήνων άνθρακα (CNTs): μονοφλοιϊκών (με ένα τοίχωμα) (Single Wall CNT: SWCNT), διπλοφλοιϊκών (με δύο τοιχώματα) (Double Wall CNT: DWCNT), πολυφλοιϊκών (με πολλαπλά (~15) τοιχώματα) (Multi Wall CNT: MWCNT), λεπτών “πολλαπλού” τοιχώματος (με λίγα (~6-7 ) τοιχώματα) (thin MWCNT), αλλά και τροποποιημένων νανοσωλήνων άνθρακα πολλαπλού τοιχώματος με υδρόξυ-ομάδες (-OH) και καρβόξυ-ομάδες (-COOH) καθώς επίσης και νανοσωλήνων άνθρακα τροποποιημένων με διάφορα πολυμερή όπως πολυβινυλοπυρολιδόνη (PVP), πολυμεθακρυλικό γλυκιδιλεστέρα (PGMA), (PSSPC16). Οι νανοσωλήνες άνθρακα, αρχικά, χαρακτηρίσθηκαν με τη βοήθεια της φασματοσκοπίας Raman και της Ηλεκτρονικής Μικροσκοπίας Σάρωσης και μελετήθηκε η διασπορά τους σε νερό (H2O) και αιθανόλη (EtOH). Κατόπιν, εμβολίσθηκαν σε διαφόρων τύπων πορώδεις ανισοτροπικές μεμβράνες (πόρων κωνικού τύπου), αλλά και σε μεμβράνες καθορισμένου μεγέθους πόρων κυλινδρικού τύπου (track etched), στην προσπάθεια ανάδειξης μιας βέλτιστης ενθυλάκωσής τους στο ενεργό/εκλεκτικό τμήμα των μεμβρανών αυτών, κάτι που δεν είναι καθόλου προφανές. Αναπτύχθηκε μια πειραματική διάταξη εμβολισμού νανοσωλήνων άνθρακα, βασιζόμενη στην αρχή της διήθησης/φιλτραρίσματος, η οποία επέτρεψε ένα βαθμό εμβολισμού τους στις μεμβράνες και μια τάση βελτίωσης του χρόνου/των ρυθμών διέλευσης του νερού από αυτές. Στην προσπάθεια αυτή αρωγός σ’ ένα μεγάλο βαθμό αποδείχθηκε η Ηλεκτρονική Μικροσκοπία Σάρωσης. / Membrane Bioreactors are an innovative technology, ideal for the treatment and rehabilitation of advanced municipal and industrial wastewater which contain high biodegradable organic compounds. A new category of functional membranes for technology MBR, which offer higher water flow, higher rejection rate of organic compounds and inorganic salts of low molecular weight, and greater resistance to the deposition of the membrane of microorganisms may be ensured by the inclusion of various types of carbon nanotubes (CNT’s) into porous polymeric membranes and its basic principle is the efficient binding of modified carbon nanotubes in these membranes. Carbon nanotubes, from the first moment of their discovery, have attracted the interest of the scientific community, due to their wide application in many scientific and technological fields, as a result of their unique properties. More specifically, the chemical, optical, electrical and mechanical properties make them potentially useful in many applications. Important is the use of carbon nanotubes for the development of an innovative high performance membrane for use in Membrane Bioreactors Technology (Membrane Bioreactors, MBR’s). In the present study different types of carbon nanotubes were examined, such as single-wall carbon nanotubes (SWCNT’s), double-wall carbon nanotubes (DWCNT’s), multi-wall carbon nanotubes (MWCNT’s), thin multi-wall carbon nanotubes (thin MWCNT’s), and modified carbon nanotubes with hydroxy groups (-OH), carboxyl groups (-COOH) as well as carbon nanotubes modified with various polymers such as polyvinylpyrrolidone (PVP), phosphonium salt of polystyrene sulfonate (PSSPC16) and polyglycidyl methacrylate (PGMA). Initially, the different types of carbon nanotubes were characterized, using Raman Spectroscopy and Scanning Electron Microscopy. Their dispersion in H2O and ethanol was also examined. Then, they were infiltrated into various types of porous anisotropic membranes with conical porous and into defined pore size membranes (track etched), to find the most suitable combination, which would result to the best water flow through the infiltrated membrane. For this purpose, an experimental device was developed, based on the principle of filtration, which allowed both the filtration of the nanotubes in the films, and the measuring of the water flow through them. Furthermore, the optimal conditions of the system were studied that could both bring about the greater coverage of the membrane pores from nanotube suspensions, (probed by SEM), and result to the optimum water flow rate.

Dissertation submitted in compliance with the requirements for the Master's Degree in Technology: Biotechnology, Technikon Natal, 2001. / The vegetable oil industry produces effluent containing quantities of fat, oil, sodium, phosphates as well as other pollutants. Oils and greases tend to clog sewers and pumps, thus creating difficulties within the municipal wastewater treatment works. Physico-chemical treatment methods, such as (Dissolved Air Flotation) OAF, gravity separation and the use of coagulants have been attempted providing a considerable reduction in organic loading; however, discharge standards are still not met. Thus, biological treatment methods are being sought after. Aerobic treatment has been attempted however, shock loads cause problems while running such a process. The objective of this study was to assess the efficiency of anaerobic digestion to degrade Vegetable Oil Effluent (VOE) as well as the efficiency of the Anaerobic Baffled Reactor (ABR). Anaerobic digestion involves the breakdown of organic matter by the action of microorganisms in the absence of oxygen, producing methane-rich biogas. The VOE was characterized, providing significant information on its chemical composition. It was found that the effluent had high sulphate content as well as a high COD content. High sulpahte content of wastewaters have known to promote growth of Sulphate Reducing Bacteria (SRB), which utilize the same energy source as Methane Producing Bacteria (MPB) and therefore compete for the same energy source. Sulphate and lipid reduction pretreatment experiments were carried out, using barium chloride and gravitational separation respectively. The results obtained, showed that the use of barium chloride to reduce sulphate content in VOE was successful, with significant sulphate reduction. The lipid reduction experiments however, did not show any significant lipid reduction. Batch tests were conducted in serum bottles to assess the extent of biodegradation of the VOE in its raw state as well as with reduced sulpahte content. Methanogenic toxicity tests on the raw and pretreated VOE provided a range of toxicity results. These assays are relatively simple and inexpensive. Gas production was monitored to determine the rate and extent of biodegradation. The efficiency of digestion was assessed by COD reduction. Results indicated potential inhibition of the methanogenic bacteria responsible for methane production by the / M

Lignocellulosic biomass is a potential feedstock for production of sugars, which can be fermented into ethanol. The work presented in this thesis proposes some solutions to overcome problems with suboptimal process performance due to elevated cultivation temperatures and inhibitors present during ethanol production from lignocellulosic materials. In particular, continuous processes operated at high dilution rates with high sugar utilisation are attractive for ethanol fermentation, as this can result in higher ethanol productivity. Both encapsulation and membrane bioreactors were studied and developed to achieve rapid fermentation at high yeast cell density. My studies showed that encapsulated yeast is more thermotolerant than suspended yeast. The encapsulated yeast could successfully ferment all glucose during five consecutive batches, 12 h each at 42 °C. In contrast, freely suspended yeast was inactivated already in the second or third batch. One problem with encapsulation is, however, the mechanical robustness of the capsule membrane. If the capsules are exposed to e.g. high shear forces, the capsule membrane may break. Therefore, a method was developed to produce more robust capsules by treating alginate-chitosan-alginate (ACA) capsules with 3-aminopropyltriethoxysilane (APTES) to get polysiloxane-ACA capsules. Of the ACA-capsules treated with 1.5% APTES, only 0–2% of the capsules broke, while 25% of the untreated capsules ruptured within 6 h in a shear test. In this thesis membrane bioreactors (MBR), using either a cross-flow or a submerged membrane, could successfully be applied to retain the yeast inside the reactor. The cross-flow membrane was operated at a dilution rate of 0.5 h-1 whereas the submerged membrane was tested at several dilution rates, from 0.2 up to 0.8 h-1. Cultivations at high cell densities demonstrated an efficient in situ detoxification of very high furfural levels of up to 17 g L-1 in the feed medium when using a MBR. The maximum yeast density achieved in the MBR was more than 200 g L-1. Additionally, ethanol fermentation of nondetoxified spruce hydrolysate was possible at a high feeding rate of 0.8 h-1 by applying a submerged membrane bioreactor, resulting in ethanol productivities of up to 8 g L-1 h-1. In conclusion, this study suggests methods for rapid continuous ethanol production even at stressful elevated cultivation temperatures or inhibitory conditions by using encapsulation or membrane bioreactors and high cell density cultivations. / <p>Akademisk avhandling som för avläggande av teknologie doktorsexamen vid Chalmers tekniska högskola försvaras vid offentlig disputation den 4 april 2014, klockan 9:30 i KE-salen, Kemigården 4, Göteborg.</p>

Biofuels production and utilisation can reduce the emission of greenhouse gases, dependence on fossil fuels and also improve energy security. Ethanol is the most important biofuel in the transportation sector; however, its production from lignocelluloses faces some challenges. Conventionally, lignocellulosic hydrolysis and fermentation has mostly been performed by separate hydrolysis and fermentation (SHF) or simultaneous saccharification and fermentation (SSF). SHF results in product inhibition during enzymatic hydrolysis and increased contamination risk. During SSF, suboptimal conditions are used and the fermenting organism cannot be reused. Bacterial contamination is another major concern in ethanol production, which usually results in low ethanol yield. In these studies, the above-mentioned challenges have been addressed. A novel method for lignocellulosic ethanol production ‘Simultaneous saccharification filtration and fermentation (SSFF)’ was developed. It circumvents the disadvantages of SSF and SHF; specifically, it uses a membrane for filtration and allows both the hydrolysis and fermentation to be carried out at different optimum conditions. SSFF also offers the possibility of cell reuse for several cultivations. The method was initially applied to pretreated spruce, with a flocculating strain of yeast Saccharomyces cerevisiae. SSFF was further developed and applied to pretreated wheat straw, a xylose rich lignocellulosic material, using encapsulated xylose fermenting strain of S. cerevisiae. High solids loading of 12% suspended solids (SS) was used to combat bacterial contamination and improve ethanol yield. Oil palm empty fruit bunch (OPEFB) was pretreated with fungal and phosphoric acid in order to improve its ethanol yield. An evaluation of biofuel production in Nigeria was also carried out. SSFF resulted in ethanol yield of 85% of the theoretical yield from pretreated spruce with the flocculating strain. Combination of SSFF with encapsulated xylose fermenting strain facilitated simultaneous glucose and xylose utilisation when applied to pretreated wheat straw; this resulted in complete glucose consumption and 80% xylose utilisation and consequently, 90% ethanol yield of the theoretical level. High solids loading of 12% SS of pretreated birch resulted in 47.2 g/L ethanol concentration and kept bacterial infection under control; only 2.9 g/L of lactic acid was produced at the end of fermentation, which lasted for 160 h while high lactic acid concentrations of 42.6 g/L and 35.5 g/L were produced from 10% SS and 8% SS, respectively. Phosphoric acid pretreatment as well as combination of fungal and phosphoric pretreatment improved the ethanol yield of raw OPEFB from 15% to 89% and 63% of the theoretical value, respectively. In conclusion, these studies show that SSFF can potentially replace the conventional methods of lignocellulosic ethanol production and that high solids loading can be used to suppress bacterial infections during ethanol productions, as well as that phosphoric acid pretreatment can improve ethanol yield from lignocellulosic biomass. / <p>Thesis for the degree of Doctor of Philosophy at the University of Borås to be publicly defended on 31 October 2014, 10.00 a. m. in room E310, University of Borås, Allégatan 1, Borås.</p>

Fruits are among the most important commodities in global trading due to its fundamental nutritional values. In 2012, the fruits supply was 115 kg/person/year, however, only 50 % of the fruits reached their consumers and the rest ended up as waste during the long fruit supply chain. The waste from fruits is mostly dumped or burned, creating a serious environmental problem. A more sustainable handling of the waste is therefore highly desirable. One of them is conversion of the fruits wastes into biogas through anaerobic digestion. One challenge with the conversion of fruits wastes into biogas is the presence of antimicrobial compounds in the fruits, which reduce the biogas yield or even cause a total failure of the process. Fruit flavors have been reported to have antimicrobial activity against several microorganisms and being responsible for the defense system in the fruits. However, there is only scarce information about the effect of fruit flavors on anaerobic digesting microbia. The objectives of the present thesis were: 1) to investigate the inhibitory activity of the fruit flavors on anaerobic digestion; 2) to remove the flavor compound by pretreatment; and 3) to protect the cell from the flavor compounds using a membrane bioreactor. The inhibitory activity of the fruit flavors was examined from different groups of flavors by adding a single flavor compound into the batch anaerobic digesting system, at three different concentrations. Among the flavors added, myrcene and octanol were found to exhibit a strong inhibitory activity, with 50 % reduction of the methane production at low concentrations, ca. 0.005–0.05 %. These flavors can be found in oranges, strawberries, grapes, plums, and mangoes. The other flavors tested showed moderate and low inhibitory activity, which might not affect the anaerobic digestion of the fruits wastes. In order to overcome the inhibitory effects of the fruit flavor, two approaches were proposed in this thesis, namely, fruit flavor removal by leaching pretreatment and cell protection from fruit flavor using a membrane bioreactor. Orange peel waste and D-limonene were used as a model of fruit waste and inhibitor, respectively. The leaching pretreatment uses solvent to extract the limonene from the orange peel. The methane yield increased by 356 % from 0.061 Nm3/kg VS to 0.217 Nm3/kg VS, by pretreating the peel using hexane with peel and a hexane ratio of 1:12 at room temperature for 10 min. Alternative to limonene removal, the cells were encased in a hydrophilic membrane, which is impermeable to hydrophobic limonene. This method yielded more than six times higher methane yield, compared to the free cell. At the highest organic loading rate, examined in this work, 3 g VS/L/day, the methane yield of the reactor containing the free cell was only 0.05 Nm3/kg VS, corresponding to 10 % of the theoretical yield, whereas 0.33 Nm3/kg VS methane yield was achieved using a membrane bioreactor corresponding to 75 % of the theoretical yield. / <p>Thesis for the degree of Doctor of Philosophy at the University of Borås to be publicly defended on November 27th 2014, 10.00 a.m. in room E310, University of Borås, Allégatan 1, Borås.</p>

Les représentants de l'ordre des Thermotogales se trouvent généralement dans les puits pétroliers et les sources hydrothermales aquatiques et terrestres. Récemment, des études moléculaires basées sur l'analyse des gènes codant l'ARNr 16S ont prouvé l'existence de représentants thermophiles, mais également mésophiles de cet ordre dans les boues de bioréacteurs et dans les sédiments contaminés par des composés toxiques. Les expériences que nous avons conduites pour traiter notamment le lactosérum en présence de phosphogypse ou de sulfate dans des bioréacteurs anaérobies mésothermiques nous ont permis de mettre en évidence de nouvelles populations de bactéries et plus précisément de Thermotogales par des approches moléculaires et culturales, avec la description de deux nouveaux genres, « Mesotoga sulfurireducens » et Defluviitoga tunisiensis. En ce qui concerne « M. sulfurireducens», il correspond à la première bactérie mésophile isolée chez les Thermotogales avec un métabolisme original centré sur la sulfo-réduction. Toujours dans le but de traiter le phosphogypse, mais cette fois-ci en utilisant les margines comme composantes organiques dans le procédé anaérobie, une nouvelle espèce du genre Fusibacter a pu être isolée, F. tunisiensis. Globalement, nos résultats démontrent le rôle important joué par les Firmicutes fermentaires et sulfato-réductrices (ordre des Clostridiales) et celui des Proteobacteria sulfato-réductrices (deltaproteobacteria) chez les Bacteria, mais également par les Methanoarchaea acétoclastes aux côtés des Thermotogales dans la digestion anaérobie de la matière organique lorsque les effluents sont riches en sulfate. / The representatives of the order Thermotogales are usually found in oil reservoirs and hot aquatic and terrestrial springs. Recently, the existence of thermophilic and mesophilic representatives of the Thermotogales was proved by analysis of SSU rRNA genes of clones from bioreactors sludges and sediments contaminated by toxic compounds.Experiments which were led to treat lactoserum in the presence of phosphogypsum or sulphate in the mesothermic anaerobic digesters permitted to detect new populations of bacteria, in particular, Thermotogales by molecular and cultural approaches, with the description of two new genera, “Mesotoga sulfurireducens” and “Defluviitoga tunisiensis”. “Mesotoga sulfurireducens” is the first mesophilic bacterium isolated in the order of Thermotogales with an original metabolism axed on sulfo-reduction. During the treatment of phosphogypsum in the presence of margines as organic compounds in the anaerobic processus, a new species of Fusibacter was isolated: F.tunisiensis, sp. nov.Our results show the important role played by fermentative and sulphate-reducing Firmicutes (order Clostridiales) and sulphate-reducing Proteobacteria (deltaproteobacteria) within the Bacteria, but also by the acetoclastic Methanoarchaea beside Thermotogales in the anaerobic digestion of organic matter when the effluents are rich in sulphate.

Les lésions de cartilage ne cicatrisent pas spontanément et la réparation de ce tissu est un challenge. Les techniques chirurgicales restant insatisfaisantes, la thérapie cellulaire et l'ingénierie tissulaire sont maintenant envisagées. La transplantation de chondrocytes autologues (TCA) existe déjà mais cette procédure nécessite l'amplification des chondrocytes qui s'accompagne d'une perte du phénotype différencié (dont l'indicateur est le collagène de type II), au profit d'un phénotype fibroblastique (dont l'indicateur est le collagène de type I, retrouvé dans les tissus fibreux). La TCA conduit donc à une greffe de chondrocytes dédifférenciés produisant un fibrocartilage, dont les propriétés mécaniques sont différentes du cartilage hyalin natif. L'objectif de mes travaux était de développer un nouveau kit d'ingénierie tissulaire du cartilage par association de chondrocytes humains, de biomatériaux et d'une sélection de facteurs solubles. Nous avons utilisé le cocktail FGF-2/insuline (FI) pour l'amplification cellulaire et le cocktail BMP-2/insuline/T3 (BIT) pour redifférencier les chondrocytes dans des éponges de collagène. Nos résultats ont montré que cette combinaison permet la synthèse d'une matrice cartilagineuse dans les supports collagène. Cependant, cette synthèse s'est trouvée favorisée en périphérie des éponges cultivées en conditions statiques. Nous avons ensuite utilisé un bioréacteur pour perfuser les éponges et nos résultats ont révélé alors un dépôt plus homogène de cartilage dans ces supports. De manière très intéressante, nous avons aussi observé l'arrêt de l'expression du collagène de type I. Ainsi, notre approche multifactorielle combinant des chondrocytes humains, des biomatériaux collagène, une combinaison FI-BIT et une culture en perfusion permet la reconstruction d'un cartilage non fibrotique / Cartilage lesions are irreversible and cartilage repair is challenging. Actual surgical techniques remain unsatisfactory and therefore, cell therapy and tissue engineering approaches are now considered. The Autologous Chondrocytes Transplantation (ACT) already exists but this procedure requires chondrocytes amplification. During this amplification, a dedifferentiation process occurs: chondrocytes lose their differentiated phenotype (characterized by type II collagen) towards a fibroblastic phenotype (characterized by type I collagen, a component of fibrous tissues). ACT leads to the graft of dedifferentiated chondrocytes, hence provoking the production of a fibrocartilage that presents different mechanical properties than native hyaline cartilage. The aim of my work was to develop a new kit of tissue engineering for cartilage repair using human chondrocytes, biomaterials and a selection of soluble factors. We used a cocktail of FGF-2 and insulin (FI) for cell amplification and a cocktail of BMP-2, insulin and T3 (BIT) for chondrocyte redifferentiation in collagen sponges. Our results showed that the combination allows the synthesis of a cartilaginous matrix in collagen scaffolds. However, matrix production is favored in periphery of the sponges cultivated in static conditions. A perfusion bioreactor was then used to perfuse the sponges and our results revealed a more homogeneous deposition of cartilage in the scaffolds. Very interestingly, we also noticed a stop of type I collagen expression. Thus, our multifactorial approach combining human chondrocytes, collagen scaffold, the combination FI-BIT and culture under perfusion allows the reconstruction of a non-fibrotic cartilage

A dissertation submitted to the Faculty of Engineering and the Built Environment, University of the Witwatersrand, Johannesburg, in fulfilment of the requirements for the degree of Master of Science in Chemical Engineering, 2016 / In South Africa, the need for water treatment is increasing, especially in the mining sector. As active water treatment technologies are expensive, the mining sector has an increasing need for passive water treatment technology, with low maintenance and operating costs, yet efficient water treatment ability. Literature on passive water treatment suggests that these systems only offer a narrow range of treatment capabilities. Therefore, hybrid water treatment systems could be a solution to low-cost water treatment in South Africa. The Degrading Packed Bed Reactor (DPBR) is one of the units comprising the hybrid treatment group. The DPBR’s main action is to convert sulfates into sulfides and alkalinity. In practice, the main drawback of the DPBR is clogging. Clogging lessens the amount of Acid Mine Drainage (AMD) that comes into contact with Sulfur Reducing Bacteria (SRB) in the DPBR, thereby reducing the efficiency of the bioreactor. In this study, six small-scale DPBRs were constructed. Each was classified according to its unique organic source (manure, straw, vegetable food processing waste, wood shavings, chicken litter and a combined sample with layers of all the carbon sources). Synthetic AMD was fed through the six bioreactors for a period of three months. From the small-scale DPBRs, the permeability, sulfate, iron and pH of the exit samples were measured. On average, the carbon sources removed 50 % of the sulfates and 98 % of the iron from the fed AMD. The different carbon sources showed no significant difference between each other in terms their sulfate and iron removal. The range between the best performing carbon source and the poorest performing carbon source, in terms of sulfate removal, was 17%. For iron removal, the range between the best and poorest performing carbon sources was only 2%. It was found that the permeability of the carbon sources played a larger role in the efficiency of the DPBR than the type of carbon source used. Manure is highly effective in terms of pH improvement, sulfate and iron removal. However, this is at the expense of permeability, as its packing clogs very rapidly. Compost and straw have excellent permeabilities which do not change significantly over long timeframes. This is, however, at the expense of the remedial ability of the packing materials. The combined reactor, in every instance, offers a good compromise between these different behaviours. / GR2016