The Upflow Anaerobic Sludge Bed reactor (UASB) has been used successfully by the food processing, pulp and paper and municipal wastewater industries. High organic strength wastewater, limited space, extended down times and sludge handling and disposal have been critical factors in waste treatment system selection. This study investigated the performance of a laboratory-scale UASB reactor for treating clam-processing wastewater. Virginia state effluent regulations for BOD₅:TSS ranged from 0:0 to 90:90 to as high as 500:500 depending on the facility location. It was found that at a volumetric organic loading rate of 13.8 g COD/L/d the BOD₅ removal efficiency averaged 87% and TSS removal efficiency averaged 83%. The average effluent values for BOD₅ and TSS were 200 ppm and 90ppm, respectively. The conversion efficiency of COD to methane gas was 81%. At an organic loading rate of 4.8 g COD/L/d the effluent values averaged 77 and 66 mg/L for BOD₅ and TSS, respectively. Methane production was inhibited at Na⁺ concentrations above 5,000 ppm in batch experiments. At Na⁺ concentrations above 12,500 ppm inhibition was essentially 100%. Nutrient enrichment did not affect methane production. / Master of Science

This study was conducted to isolate and characterize bacteria that have the capacity to degrade both carbofuran and dicamba. The pathways of degradation for both pesticides were elucidated. An aerobic, carbofuran-degrading bacterium was isolated from a high concentration carbofuran bioreactor. The isolate degrades carbofuran at the upper limit of carbofuran solubility (approximately 700 mg L⁻¹), to carbofuran phenol. In aqueous mineral salts medium with carbofuran as Furadan 4F (6 g L⁻¹ a.i.), degradation of carbofuran to undetectable levels required approximately 100 days. Although carbofuran phenol was not completely degraded, the cells remained viable in the presence of unusually high concentrations of both surfactant and carbofuran phenol. Additional nutrient sources had little effect upon the rate of degradation of carbofuran in pure culture. A dicamba-degrading consortium enriched from wetland soil, using the batch culture method, was used to elucidate the pathway of dicamba degradation under anaerobic conditions. The consortium consisted of one sulfate reducing bacterium, one fermenter, and three methanogens. The sulfate reducing bacterium was isolated from the consortium using sulfate as a terminal electron acceptor and 2-bromoethanesulfonic acid was added to inhibit the growth of the methanogens. Since the fermenter is dependent upon the methanogens, elimination of these organisms caused the elimination of the fermenter. Three methanogens (Methanothrix, Methanosarcina and Methanospirillum sp) were isolated with acetate and headspace gas consisting of H₂-CO₂. Degradation of dicamba proceded through an initial demethylation reaction yielding 3,6-dichlorosalicylic acid, as determined by high performance liquid chromatography (HPLC) analysis of aqueous medium. This was followed by a reductive dehalogenation reaction at the meta position of 3,6-dichlorosalicylic acid forming 6-chlorosalicylic acid. The metabolites were isolated using thin layer chromatography. Confirmation of metabolite identity was achieved using HPLC, and mass spectrometry. It appears that the fermenter was responsible for mediating the demethylation reaction. The consortium was unable to mineralize the aromatic ring. The substrate specificity of the dicamba-degrading consortium was investigated. The consortium was found to have the capacity to mediate the reductive dehalogenation of both 3-chlorosalicylic acid and 2,5-dichlorobenzoic acid at the meta position. The consortium was unable to dehalogenate either 3-chlorobenzoic acid, 4-chlorosalicylic acid, 5-chlorosalicylic acid, or 2,5-dichlorophenol. Addition of the reducing agent cysteine (0.025% and 0.050%) to a yeast extract amended (0.04%) mineral salts medium containing 3-chlorosalicylic acid reduced the rate of dehalogenation compared to medium containing sodium sulfide as the reducing agent. Only limited dehalogenation of 3- chlorosalicylic acid and 2,5-dichlorobenzoic acid was observed when the sulfate reducing bacterium was cultured alone in a yeast extract amended medium, suggesting that the mutualistic efforts of a mixed population of anaerobes were necessary to efficiently mediate reductive dehalogenation. / Ph. D.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnology, Durban Institute Of Technology, 2005 xxiii, 172 p. : ill. ; 30 cm / The provision of safe and sanitary water is a constitutional right and above all, a necessity of life. As a result of the rapid urbanisation and the past policies of apartheid, a large population of South Africa dwell in informal settlements, where there is very little hope of development, as the government does not possess the resources that are necessary for a full-scale sanitation programme. Therefore, on-site treatments have been considered to provide sanitation in these dense peri-urban areas. The anaerobic baffled reactor (ABR) is one such sanitation system. This reactor utilises the phenomenon of anaerobic digestion to degrade substrates. One of the major disadvantages of any anaerobic treatment processes is the extreme sensitivity of the bacterial communities, thus inducing slow recovery rates following toxic shocks. Therefore, an understanding of these microbial consortia is essential to effectively control, operate and optimise the anaerobic reactor. Fluorescence in situ hybridization, 4’,6-diamidino-2-phenylindole (DAPI) staining and DNA sequencing techniques were applied to determine the microbial consortium, as well as their reactions to daily operating conditions. With an understanding of these populations and their responses to perturbations within the system, it is possible to construct an anaerobic system that is successful in its treatment of domestic wastewater. In situ hybridizations were conducted for three operating periods, each characterised by specific flow rates. Results showed Eubacterial population dominance over the Archaeal population throughout both of the operating periods investigated. However, these cells cumulatively consisted of 50% of the total biomass fraction, as determined by DAPI staining. Group-probes utilised revealed a high concentration of fermentative acidogenic bacteria, which lead to a decrease in the pH values. It was noted that the ABR did not separate the acidogenic and methanogenic phases, as expected. Therefore, the decrease in pH further inhibited the proliferation of Archaeal acetoclastic methanogens, which were not present in the second operating period. DNA sequencing results revealed the occurrence of the hydrogenotrophic Methanobacterium and Methanococcus genera and confirmed the presence of Methanosarcina. Sequencing of the bacterial DNA confirmed the presence of the low G+ C Gram Positives (Streptococcus), the high G+C Gram Positives (Propionibacterium) and the sulfate reducing bacteria (Desulfovibrio vulgaris). However, justifications were highly subjective due to a lack of supportive analytical data, such as acetate, volatile fatty acids and methane concentrations. Despite this, findings served to add valuable information, providing details on the specific microbial groups associated with ABR treatment processes.

Thesis (MSc)--Stellenbosch University, 2000. / ENGLISH ABSTRACT: A severely polluted industrial effluent is generated by the local gelatinmanufacturing industry. Due to increasingly stringent restrictions on discharge qualities enforced by the National Water Act of 1998 and National Environmental Management Act of 1998, as well as increasing trade-effluent charges implemented via the Local Municipal Bylaws, the industry is compelled to consider a system to pre-treat the polluted effluent. A study was undertaken to examine the viability of anaerobic treatment of the gelatin-manufacturing effluent, since the anaerobic digestion technology is well recognised for the high success rate in the treatment of high-strength, complex wastewaters. Various laboratory and pilot-scale studies were done, using different hybrid Upflow Anaerobic Sludge Blanket (UASB) and contact designs. Two mesophilic laboratory-scale hybrid UASB digester designs, fitted with polyethylene (AD-1) and polyurethane (AD-2), performed well at a hydraulic retention time (HRT) of 1.0 d. Chemical oxygen demand (COD) removal efficiencies of up to 90% (avg. 53%) for AD-1 and 83% (avg. 60%) for AD-2 at organic loading rates (OLR) of 9.56 and 4.62 kg COD.m-3.d-1, respectively, were obtained. High sulphate (S04) removal efficiencies of up to 96% (avg. 86%) for AD-1 and 98% (avg. 82%) for AD-2 were also achieved, respectively. A maximum total solid (TS) removal of 65% (avg. 25%) for AD-1 and 62% (avg. 28%) for AD-2 was reported. An average methane content of 80% (AD-1) and 79% (AD-2) with average methane yields per COD removed of 2.19 and 1.86 m3. kg CODremoved.df-o1r AD-1 and AD-2 were found, respectively. When the same digesters (AD-1 and AD-2) were combined in a muItiphase series configuration, a total COD removal efficiency of up to 97% (avg. 80%) at an OLR of 8.32 kg COD.m-3.d-1,was achieved. Excellent total S04 removals of 96% (avg. 69%) were accomplished. Up to 82% TS (avg. 29%) was also removed during this study and the biogas consisted of 89% methane (avg. 79%). For this multi-phase combination up to 92% volatile fatty acids (VFA) (avg. 48%) were removed, indicating possible selective phase separation of the respective fatty acid producing/utilising bacterial populations. The use of a laboratory-scale UASB bioreactor with recirculation, resulted in COD removal efficiencies of up to 96% (avg. 51%) at an HRT of 3.0 d, and 95% (avg. 54%) at a HRT of 1.0 d. Low performances were generally found, with average S04 and TS removals of 59% (max. 97%) and 26% (max. 67%), respectively at an HRT of 1.0 d. The biogas production was very low throughout the study (0.05 - 0.63 I,d-1 ). A pilot-scale UASB reactor (300 I) was constructed and performed satisfactory with a 58% average COD removal and maximum of 96%. S04 and TS removals up to 96% (avg. 44%) and 93% (avg. 63%), respectively, were obtained. The methane content of the biogas was 85%. The pilot-scale studies were conducted under actual field conditions, where various shock and organic loads had to be absorbed by the system. The pilot-scale contact configuration (300 I) did not perform satisfactory as a result of continuous blockages experienced in the feed and recirculation lines. Maximum COD, S04, VFA and TS removal efficiencies of 41% (avg. 27%), 62% (avg. 41%), 64% (avg. 27%) and 39% (avg. 21%), respectively, were obtained. The results of all the studies indicated acceptable COD removals with increasing OLR's. Indications of the presence of active methanogenic and sulphate-reducing bacterial populations were apparent throughout the studies. One possibility for the successful start-up and commissioning of the anaerobic reactors was the use of a well-adjusted biomass, which consisted of highly selected and adapted microbial consortium for the specific gelatinmanufacturing effluent. It was clear from this study that gelatin-manufacturing effluent can be treated successfully, especially with the use of the UASB design. A welldefined data base was constructed which could be of great value for further upscaling to a full-scale digester. / AFRIKAANSE OPSOMMING: 'n Hoogs besoedelde industriele uitvloeisel word gegenereer deur die plaaslike gelatien-vervaardigings industrie. As gevolg van toenemende streng beperkings op die kwaliteit van uitvloeisels wat bepaal word deur die Nasionale Water Wet van 1998 en Nasionale Omgewings Bestuurs Wet van 1998, asook toenemende munisipale heffings wat geimplementeer word via Plaaslike Munisipale Wette, word die industrie verplig om die uitvloeisel vooraf te behandel. 'n Studie is onderneem om die lewensvatbaarheid van anaërobe behandeling van gelatien-vervaardigings uitvloeisel te ondersoek, aangesien anaërobe verterings tegnologie alombekend is vir die goeie sukses behaal in die behandeling van hoë-sterkte, komplekse uitvloeisels. Verskeie laboratorium- en loods-skaal studies is gedoen, met verskillende hibried Opvloei Anaërobe Slykkombers (OAS) en kontak ontwerpe. Goeie werksverrigting was verkry by 'n hidroliese retensie tyd (HRT) van 1.0 d met twee mesofiliese laboratorium-skaal hibried OAS verteerder ontwerpe wat uitgevoer was met poli-etileen (AD-1) en poli-uretaan (AD-2) materiaal. Chemiese suurstof behoefte (CSB) verwyderings van so hoog as 90% (gem. 53%) vir AD-1 en 83% (gem. 60%) vir AD-2 by organiese ladingstempo's (OLT) van 9.56 en 4.62 kg CSB.m-3.d-1,was onderskeidelik verkry. Hoë sulfaat (S04) verwyderings van tot 96% (gem. 86%) vir AD-1 en 98% (gem. 82%) vir AD-2 was ook onderskeidelik verkry. 'n Maksimum totale vaste stof (TVS) verwydering van 65% (gem. 25%) vir AD-1 en 62% (gem. 28%) vir AD-2 is gerapporteer. 'n Gemiddelde metaan inhoud van 80% (AD-1) en 79% (AD-2) met 'n gemiddelde metaan opbrengs per CSB verwyder van 2.19 en 1.86 m3.kg CSBverwyder.dv-i1r AD-1 en AD-2, was onderskeidelik gevind. Met die aanwending van dieselfde twee verteerders (AD-1 en AD-2) in 'n series gekoppelde multi-fase konfigurasie, is 'n totale CSB verwydering so hoog as 97% (gem. 80%) verkry by 'n OLT van 8.32 kg CSB.m-3.d-1. Uitstekende totale S04 verwydering van 96% (gem. 69%) is behaal. Tot 82% TVS (gem. 29%) was vewyder gedurende die studie en die biogas het uit 89% metaan (gem. 79%) bestaan. Vir die multi-fase kombinasie is 'n maksimum van 92% vlugtige vetsure (WS) (gem. 48%) verwyder, wat dui op die moontlike skeiding van selektiewe fases van die onderskeie vetsuur produserende/verbruiker bakteriële populasies. CSB verwydering van tot 96% (gem. 51%) by 'n HRT van 3.0 d en 95% (gem. 54%) met 'n HRT van 1.0 d was verkry, tydens die gebruik van In laboratorium-skaal OAS bioreaktor met hersirkulasie. Lae werksverrigting was oor die algemeen waargeneem, met gemiddelde S04 en TVS verwyderings van 59% (maks. 97%) en 26% (maks. 67%) by In HRT van 1.0 d. Die biogas produksie was baie laag gedurende die studie (0.05 - 0.63 I,d-\ In Loods-skaal OAS verteerder was opgerig en bevredigende resultate was verkry met In gemiddeld van 58% CSB verwydering en maksimum van 96%. S04 en TVS verwyderings so hoog as 96% (gem. 44%) en 93% (gem. 63%) is onderskeidelik verkry. Die metaan inhoud van die biogas was 85%. Die loods-skaal studie was uitgevoer gedurende ware veld kondisies, waartydens verskeie skok en organiese ladings deur die sisteem geabsorbeer is. Die loods-skaal kontak konfigurasie (300 I) het nie bevredigende resultate getoon nie, as gevolg van voortdurende blokkasies wat ondervind is in die toevoer en hersirkulasie pype. Maksimum CSB, S04, WS en TVS verwyderings van 41% (gem. 27%), 62% (gem. 41%), 64% (gem. 27%) en 39% (gem. 21%) was onderskeidelik verkry. Die resultate van al die studies het aanvaarbare CSB verwydering aangedui by toenemende OLT's. Indikasies van aktiewe metanogene en sulfaat-reduserende bakteriële populasies was ook teenwoordig gedurende die studies. Die suksesvolle aansit-prosedure en begin van die anaërobe verteerders kan toegeskryf word aan die gebruik van In goed aangepaste biomassa, wat uit hoogs selektiewe en aangepaste mikrobiese populasies vir die spesifieke uitvloeisel bestaan. Hierdie studie het getoon dat gelatien-vervaardigings uitvloeisel suksesvol met die OAS ontwerp behandel kan word. In Goed gedefinieerde data basis kan voorsien word, wat van groot waarde sal wees vir verdere opgradering na In volskaalse verteerder.

Thesis (MSc)--Stellenbosch University, 2011. / ENGLISH ABSTRACT: Fermentation in Zymomonas mobilis has been described as a catabolic highway, with 50 % of soluble protein comprising glycolytic and fermentative enzymes. In conjunction with one of the fastest observed fermentations, the conversion of glucose to ethanol forms one of the least efficient energy extractions found in nature. The low energy yield of fermentation in Z. mobilis is a result of the usage of the Entner-Doudoroff glycolytic pathway, which has half the energy yield per mol substrate compared to the well known Embden-Meyerhof-Parnas glycolytic pathway. The work presented in this thesis forms part of a larger project to compare glycolytic regulation in different micro-organisms (i.e., Z. mobilis, Escherichia coli, Saccharomyces cerevisiae and Lactococcus lactis). These organisms were chosen based on their usage of different glycolytic mechanisms. By using supply-demand analysis for quantifying glycolytic regulation as well as similar experimental conditions (e.g. using non-growing cell cultures), we can compare the regulatory behaviour of mechanistically distinct freeenergy supplies. The aim of this thesis was to quantify the importance of anaerobic free-energy generation for the regulation of the Entner-Doudoroff glycolytic pathway in Z. mobilis. We used metabolic control analysis (MCA) and supply-demand analysis to realize this goal. The central message of MCA is that when a metabolic parameter (e.g., a conserved metabolic moiety) is deemed important for affecting a particular steady-state variable (i.e., fermentation flux), its effect on the steady state variable should be tested. An extension to MCA, supply-demand analysis, provides a quantitative framework for analyzing the regulatory importance of cellular commodities such as anaerobic free-energy. This is done through comparing the elasticities of anaerobic free-energy supply and demand, which yields the degree to which the respective reaction blocks control the flux through anaerobic free-energy metabolism, as well as determine the cellular free-energy state (ATP/ADP ratio). The regulation of anaerobic free-energy metabolism in Z. mobilis was investigated with an experimental approach. The key features of our experimental setup were the use of NMR spectroscopy for detecting metabolites, as well as employing non-growing conditions for supply-demand experiments. With NMR spectroscopy metabolites could be detected in real time without using invasive sampling techniques; the use of nongrowing conditions further simplified the analysis by enabling us to correlate fermentative behaviour exclusively with the anaerobic free-energy state. Fermentation of glucose was investigated in the wild type Z. mobilis, a recombinant containing a non-expressing plasmid, or expressing plasmids for over-expressing the glucose facilitator (TCDB 2.A.1.1.4) or glucose-6-phosphate dehydrogenase (EC 1.1.1.49). In addition, ATP demand in the non-expressing recombinant and wild type was perturbed by titrating with the uncoupler acetic acid. Our results show that the anaerobic free-energy demand, the glucose facilitator and glucose-6-phospate dehydrogenase all control the flux of ethanol production in Z. mobilis. The Entner-Doudoroff glycolytic supply activity was found to be sensitive to changes in the ratios of ATP/ADP (elasticity varied between –0.31 and –0.49) and NTP/NDP (elasticity varied between –0.31 and – 0.50). / AFRIKAANSE OPSOMMING: Fermentasie in Zymomonas mobilis word beskryf as ‘n kataboliese snelweg, waar glikolitiese en fermentatiewe ensieme 50% van totale oplosbare proteïene in die sel uitmaak. Hoewel dié fermentasie een van die vinnigstes is wat tot op hede waargeneem is, is die omskakeling van glukose na etanol een van die mees ondoeltreffende energieekstraksies in die natuur. Dié lae energie-opbrengs, soos waarneembaar in fermentasie in Zymomonas mobilis, kan toegeskryf word aan die Entner-Doudoroff metaboliese pad. Hierdie metaboliese pad lewer slegs die helfte van die energie-opbrengs per mol substraat vergeleke met die meer bekende Embden-Meyerhof-Parnas glikolitiese pad. Die navorsing in hierdie tesis is deel van ‘n omvattende projek wat poog om die regulering van glikolise in verskillende mikro-organismes (Z. mobilis, Escherichia coli, Saccharomyces cerevisiae en Lactococcus lactis) te vergelyk. Dié organismes is gekies op grond van die uiteenlopende glikolitiese meganismes waarvan hulle gebruik maak. Ten einde die reguleringsgedrag van meganisties verskillende vry-energie produksieweë m.b.v. vraag-aanbod analise te vergelyk, moet glikolitiese regulering eers onder eenderse eksperimentele kondisies (b.v. nie-groeiende selkulture) gekwantifiseer kan word. Die hoofdoel van hierdie tesis was om die belang van anaerobiese vry-energie produksie vir die regulering van die Entner-Doudoroff glikolitiese pad in Z. mobilis te kwantifiseer. Hiervoor is van Metaboliese kontrole-analise (MKA) en vraag-aanbodanalise (‘n uitbreiding van MKA) gebruik gemaak. MKA is ‘n tegniek waarmee die effek wat ‘n metaboliese parameter (soos metaboliese deel-konservering) op ‘n spesifieke bestendige toestand-veranderlike (soos fermentasiefluksie) het, gekwantifiseer kan word. Vraagaanbodanalise daarenteen, bied ‘n kwantitatiewe raamwerk waardeur die regulatoriese belang van sellulêre kommoditeite (byvoorbeeld anaerobiese vry-energie) geanaliseer kan word. Tydens laasgenoemde proses word die elastisiteit van die anaerobiese vry-energie aanbod en die elastisiteit van die vraag vergelyk. Op hierdie manier kan die mate van beheer wat die onderskeie reaksieblokkie oor die fluksie deur anaerobiese vry-energie metaboliese paaie, sowel as oor die sellulêre vry-energie toestand (ATP/ADP verhouding), bepaal word. In hierdie werk is die regulering van anaerobiese vry-energie metabolisme in Z. mobilis ondersoek deur van ‘n eksperimentele benadering gebruik te maak. Die sleuteleienskappe van dié benadering was om kernmagnetiese-resonansiespektroskopie (KMR spektroskopie) te gebruik om metabolietkonsentrasies te meet, en om van niegroeiende kondisies gebruik te maak vir die vraag-aanbod eksperimente. Metabolietkonsenstrasies kon aaneenlopend bepaal word sonder die gebruik van monsternemingstegnieke wat die reaksie sou kon beïnvloed. Eksterne invloede op die fermentasiegedrag kon ook uitgesluit word deur van nie-groeiende kondisies gebruik te maak, sodat die waargenome fermentasiegedrag uitsluitelik aan die anaerobiese vryenergie toestand toegeskryf kan word. Glukose fermentasie was ondersoek in wilde tipe Z. mobilis, en in drie rekombinante wat onderskeidelik ‘n glukose fasiliteerder ooruitdrukkingsplasmied (TCDB 2.A.1.1.4), ‘n glukose-6-fosfaat dehidrogenase ooruitdrukkingsplasmied (EC 1.1.1.49), en ‘n nieuitdrukkingsplasmied bevat het. Die ATP vraag in die wilde tipe en die nieuitdrukkingsrekombinant is geperturbeer deur titrasies met asynsuur as ontkoppelaar. Die resultate toon dan die anaerobiese vry-energievraag, sowel as die glukose fasiliteerder en glukose-6-fosfaat dehidrogenase, die fluksie van etanolproduksie in Z. mobilis beheer. Die Entner-Doudoroff glikolitiese produksie-aktiwiteit was sensitief vir veranderinge in die ATP/ADP verhouding (elastisiteite was tussen -0.31 en -0.49) en die NTP/NDP verhouding (elastisiteite was tussen -0.31 en -0.50).

published_or_final_version / Civil Engineering / Doctoral / Doctor of Philosophy

The aim of the investigation was to compare the performance of an anaerobic baffled reactor (ABR) with a completely mixed anaerobic reactor (CMAR). The ABR was operated with a hydraulic retention time (HRT) of 20 h while the CMAR was operated at 20 d. A control experiment was conducted with a CMAR operated at a constant hydraulic retention time and substrate feed concentration. During the first phase, the start-up performance of the ABR and CMAR were compared. In the second phase of the study the steady state COD removals were compared. The laboratory completely mixed anaerobic reactor was a 20L glass vessel with a stirrer coming in through the neck. A second type of reactor, anaerobic baffled reactor (ABR) was also operated. The ABR was a rectangular perspex box with internal vertical baffles alternately hanging and standing. The baffles divide the reactor into eight compartments with a total working volume of 7.5 L. Each baffle is angled at about 45Q to distribute the flow towards the centre of the upcomer. The reactors were seeded with raw sewage and allowed to stand for 3 days after which a continous feed of sucrose and basal salts was commenced. The initial HRT for the ABR and the CMAR were 60 h and 30 days respectively. When the reactors reached steady state (pH, gas production, gas composition and alkalinity), the HRT was reduced in a stepwise fashion (ABR 60 h to 35 h to 20 h and CMAR 35 d to 30 d to 20 d). At the final HRT the COD removals were similar (67 %). The ABR took 120 d to attain final steady state while the CMAR took 200 d. The organic loading tests were undertaken with a stepwise increase (doubling) in the influent substrate concentration. The feeding commenced at an organic loading rate (OLR) of 4.8 kg/m(3).d for the ABR. The flow rate (HRT) into both reactors and other parameters were kept constant (HRT of 20 h and 20 d for ABR and CMAR respectively). The substrate concentration was increased from 4 gCOD/L (4.8 kg/m(3).d) to 64 gCOD/L (76.8 kg/m(3).d) for the ABR. For the CMAR it was increased from 4 gCOD/L (0.25 kg/m(3).d) to 32 gCOD/L (2 kg/m(3).d). The method used was to increase the organic loading rate until the reactors failed. Since the two reactors had different operating HRTs, the tests began when both had the same COD removal rate of about 60 % COD reduction. The same parameters as in the start-up period were monitored for both reactors. The CMAR had a COD removal efficiency ca. 70 %, which did not fluctuate when OLR was increased. The ABR reached a maximum COD removal of 80 %. An increase in the OLR led to an initial decrease in the COD removal until the biomass recovered and the high COD (80 %) removal rates resumed. The ABR reached a maximum OLR of 76.8 kg/m(3).d whilst the CMAR reached a maximum OLR of 2.0 kg/m(3).d. The investigations showed that the ABR could be operated at higher organic loads than the CMAR and give the same organic removal rate. This verified the importance of increasing the SRT/HRT ratio in anaerobic reactors. The CMAR, however, proved to be stable to changes in the influent feed strength, as there was no immediate noticeable changes in the gas production. / Thesis (M.Sc.Eng.)-University of Natal, Durban, 2000.

Preterm birth, defined at birth before 37 weeks gestation, affects millions of newborns worldwide every year. Preterm birth is a leading cause of infant morbidity and mortality. One major cause of preterm birth is preterm premature rupture of membranes (PPROM), which can be triggered by bacterial infection and inflammation. A bacterial species that has been implicated in preterm birth and other obstetric complications is Sneathia amnii. The goals of this study were to observe cytopathogenic effects caused by S. amnii strain Sn35 and identify putative virulence factors causing those effects. Sn35 was able to adhere to, invade, and damage/kill various host cell lines. We characterized these virulence attributes. A putative virulence determinant was identified, and a fragment of the protein was expressed for polyclonal antiserum production. Antiserum was used to characterize the expression and subcellular localization of the protein in Sn35. However, antiserum was unable to prevent cytopathogenic effects.

Orientador: Luciane Dias de Oliveira / Banca: Luis Felipe das Chagas e Silva de Carvalho / Banca: Jônatas Rafael de Oliveira / Resumo: As plantas medicinais e os fitoterápicos têm sido utilizados como coadjuvantes e alternativos no combate a diversas doenças com crescente frequência, no entanto, na Odontologia seu uso ainda é bastante limitado. Com isso, o objetivo deste estudo é avaliar a ação antimicrobiana dos extratos aquoso e glicólico de própolis verde sobre micro-organismos anaeróbios de interesse odontológico, bem como sua citotoxicidade, a fim de introduzir e incentivar o uso efetivo e sistemático desse fitoterápico em produtos como dentifrícios e enxaguatórios bucais no combate a cáries e doenças periodontais. Os extratos comerciais aquoso e glicólico de própolis foram obtidos das empresas Apis Flora e Mapric, respectivamente. Para avaliação da atividade antimicrobiana foram utilizadas cepas-padrão (ATCC) de Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis Porphyromonas gingivalis e Prevotella intermedia em cultura planctônica, verificando a concentração inibitória mínima e concentração microbicida mínima (CIM e CMM), segundo Clinical and Laboratory Standards Institute. Para biofilmes monotípicos, suspensões padronizadas (107 céls/mL) foram adicionadas em poços de microplacas e após 48 h em anaerobiose foram tratados com 3 concentrações do extrato de própolis (n=12) por 5 min. Foi incluído um controle positivo (solução fisiológica) e um controle negativo (clorexidina). O biofilme foi mensurado pelos testes MTT e Cristal Violeta. Para análise de citotoxicidade, queratinócitos hu... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Medicinal plants and herbal medicines have been used as adjuvants and alternatives in fighting various diseases with increasing frequency. However, in dentistry its use is still quite limited. Therefore, the objective of this study is to evaluate the antimicrobial action of the aqueous and glycolic extracts of green propolis on anaerobic microorganisms of dental interest, in order to introduce and encourage the effective and systematic use of this herbal medicine in products such as dentifrices and mouthwashes in the fight against tooth decay and periodontal diseases. Aqueous and glycolic commercial extracts of propolis were obtained from Apis Flora and Mapric companies, respectively.To evaluate the antimicrobial activity, standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella intermedia in planktonic culture was used, verifying the minimum inhibitory concentration and minimum microbicide concentration (MIC and CMM), according to Clinical and Laboratory Standards Institute. For monotypic biofilms, standardized suspensions (107 cells / mL) was added to microplate wells and after 48 h in anaerobiosis was treated with 3 concentrations of propolis extracts (n = 12) for 5 min. A positive control (saline solution) and a negative control (chlorhexidine) was included. The biofilm was measured by the MTT and Violet Crystal tests. For cytotoxicity analysis, human keratinocytes (HaCaT) were cultured and ... (Complete abstract click electronic access below) / Mestre

Orientador: Luciane Dias de Oliveira / Banca: Silvana Soléo Ferreira dos Santos / Banca: Graziella Nuernberg Back-Brito / Banca: Luana Marotta Reis de Vasconcellos / Banca: Jônatas Rafael de Oliveira / Resumo: A resistência microbiana aos antibióticos disponíveis é preocupação constante, devido à dificuldade no tratamento de infecções causadas por cepas resistentes, em decorrência do uso indiscriminado de antimicrobianos. Assim, a busca por terapias antimicrobianas alternativas tem sido crescente e necessária, sendo a fitoterapia umas das opções de escolha. O objetivo do presente estudo foi analisar a atividade antibacteriana de extratos glicólicos de Achyrocline satureioides (macela), Cynara scolymus (alcachofra), Hamamelis virginiana (hamamelis) e Persea americana (abacateiro), pelos períodos de 5 min e 24 h de exposição sobre bactérias anaeróbias Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas endodontalis e Porphyromonas gingivalis, em culturas planctônica e biofilmes. As bactérias armazenadas a -80ºC foram ativadas em caldo Brucella enriquecido (hemina 1%, menadiona 1% e sangue de carneiro desfibrinado 5%) e incubadas em câmara de anaerobiose por 48 h a 37ºC por sete dias. A partir de culturas puras, o teste de microdiluição em caldo foi conduzido em microplacas por meio de suspensões bacterianas padronizadas em solução fisiológica estéril (NaCl 0,9%) e diluições dos extratos em caldo, sendo as placas incubadas por 48 h a 37ºC em anaerobiose. Alíquotas de cada poço foram semeadas em ágar Brucella enriquecido. Após incubação, a Concentração Inibitória Mínima (CIM) e Concentração Bactericida Mínima (CBM) foram determinadas. As concentrações efetivas de ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Microbial resistance to antibiotics available is constant concern, due to the difficulty in treating infections caused by resistant strains as a result of the indiscriminate use of antimicrobials. Thus, the search for antimicrobial alternative therapies has been growing and necessary, being one option the herbal medicine. The objective of the present study was to analyze the antibacterial activity to Achyrocline satureioides glycolic extracts (macela), Cynara scolymus (artichoke), Hamamelis virginiana (Witch-Hazel) and Persea americana (avocado), for periods of 5 min and 24 h from exhibition on anaerobic bacteria Fusobacterium nucleatum subsp. nucleatum, Parvimonas micra, Porphyromonas gingivalis and Porphyromonas endodontalis in planktonic communities and biofilms. Bacteria stored at -80°C have been activated in Brucella broth enriched (hemin 1%, menadione 1% and defibrinated sheep blood 5%) and incubated in anaerobiose chamber for 48 h at 37° C for seven days. From pure cultures, the microdiluição test in broth was conducted in microplates through standardized bacterial suspensions in sterile saline solution (NaCl 0.9%) and dilution of the extracts in broth, being incubated plates for 48 h at 37° C in anaerobiosis. Aliquots of each well were sown in Brucella agar enriched. After incubation, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) were determined. Effective concentrations of each extract were applied on the biofilms of each spe... (Complete abstract click electronic access below) / Doutor