Les résultats de cette étude de la composition du fruit à pain récolté sur un territoire contrasté du point de vue agropédoclimatique, et de son amidon, montrent que la teneur en amidon du fruit à pain apparaît comme étant influencée par la saison, mais pas par la zone de récolte. La teneur en amidon beaucoup plus élevée pour les fruits à pain développés durant la saison la plus sèche est un résultat très original. La connaissance du cycle de développement des fruits à pain à la Martinique a été décrite pour une première fois, permettant de déterminer une période de récolte optimisée par rapport à la floraison : la 16ème semaine pour une croissance maximale, ou la 17ème semaine pour une plus haute teneur en amidon. L’étude de Worrell et al., (1998), la seule publiée actuellement, menée à la Barbade, a montré un cycle plus long de 2 semaines environ.Du point de vue de la diversité génétique, les fruits à pain récoltés en Martinique constituent un seul groupe, à l’opposé de ceux récoltés en Guadeloupe qui constituent 2 sous-groupes proches.En ce qui concerne l’amidon, nos résultats montrent que les dimensions des grains d’amidon de fruits matures sont comprises entre 9 et 12 µm. Leur taille augmente avec la croissance du fruit puisque dans un fruit immature leur diamètre moyen était de 7,5 µm. Il s’agit d’amidons de type B à très haute cristallinité, dont les températures d'empesage sont autour de 75 °C. Parmi les facteurs agropédoclimatiques, l’humidité a eu un rôle actif sur la qualité de l’amidon puisque à la fois la quantité d’amylose et la masse molaire de l’amylopectine diminuent. L’amidon a une solubilité et un gonflement faible du même ordre de grandeur que les amidons classiques et une forte viscosité à la gélatinisation et à la rétrogradation, lui conférant une aptitude à l’utilisation comme gélifiant.Des essais d’applications alimentaires ont été effectués en panification et en extrusion. Des transformations en produits de type crackers ont donné des résultats moyennement satisfaisant avec 100 % de farine de fruit à pain. En panification l’incorporation de farine de fruit à pain a des actions délétères sur la valeur boulangère. L’acceptabilité d’un goût « nature, vert » conféré par la farine de fruit à pain devra notamment être évaluée lors de travaux ultérieurs.Le développement de produits à base de fruit à pain permettra / The results of this study the composition of breadfruit harvested in an area of agro climatic soil contrasting views, and its starch, show that the starch content of breadfruit appears to be influenced by season, but not by collection area. The starch content is much higher for breadfruit developed during the driest season is a very original result. Knowledge of the development cycle of breadfruit in Martinique has been described for the first time, to determine an optimal harvest period compared to flower: the 16th week for maximum growth, or the 17th week for a higher starch content. The study by Worrell et al., (1998), the only currently published, conducted in Barbados, showed a longer cycle of about 2 weeks.From the perspective of genetic diversity, breadfruit harvested in Martinique as a single group, in contrast to those collected in Guadeloupe that are close to two subgroups.With regard to starch, our results show that the size of starch granules of mature fruits are between 9 and 12 microns. Their size increases with the growth of the fruit as an immature fruit in their mean diameter was 7.5 microns. This is B-type starches with very high crystallinity, with temperatures of gelatinization are around 75 °C. Agro climatic soil factors, humidity took an active role on the quality of starch, since both the amount of amylose and amylopectin molecular weight of the decline. The starch has a solubility and low swelling of the same order of magnitude as the conventional starches and high viscosity to gelatinization and retrogradation, giving it a suitability for use as a gelling agent.Testing of food applications were made in bakery and extrusion. Changes in products like crackers gave moderately satisfactory results with 100 % breadfruit flour. Incorporation into bread flour breadfruit has deleterious actions on the baking. The acceptability of a taste "nature green" conferred by the breadfruit flour will be particularly assessed in future work.Product development based breadfruit allow several things, which meet the growing demand for food diversification.

La préservation de la richesse aquatique est devenue un souci majeur d'ordre mondial suite au risque de pénurie en eau. Au Liban, les rejets anthropiques incontrôlés et incontrôlables au bord des rivières menacent la qualité de ses eaux de surface. Pour cela, quatre cours d'eau libanais ont été choisis comme cadre d'étude : la rivière Damour, la rivière Kadisha-Abou Ali, la rivière Ibrahim et la rivière Oronte.Les différents paramètres physico-chimiques et microbiologiques étudiés ont permis dans un premier temps la construction d'une base de données de chacune de ces rivières. Une seule analyse spatio-temporelle des paramètres séparément n'aide pas à définir l'état trophique des rivières. Partant de l'idée des corrélations que peuvent exister entre certains paramètres, l'Analyse en Composante Principale (ACP) retenant la totalité de l'information sera utilisée en dépit des méthodes classiques. Cet outil statistique a permis de classer les rivières par niveau de pollution. Il a aussi aidé à observer l'impact des apports des polluants sur les différentes stations de rivières étudiées.Pour suivre le devenir des coliformes fécaux dans les eaux, l'ACP des variables microbiologiques a montré la persistance des colonies bactériennes dans les eaux malgré les conditions climatiques diverses et le régime méditerranéen torrentiel des rivières. En effet, les sédiments constituent des réservoirs potentiels de microorganismes pathogènes. Le décrochage bactérien des agrégats sédimentaires et la remise en suspension dans l'eau pose un problème alarmant de santé publique. / Preservation of aquatic wealth has worldwide become a major concern due to the risk of water shortage. In Lebanon, uncontrolled and uncontrollable anthropogenic rejections along rivers threaten the quality of its surface waters. Four Lebanese rivers were selected as study framework: The Damour river, Kadisha-Abu Ali river, Ibrahim river and Orontes river. The physico-chemical and microbiological parameters analyzed has formed a large database of these rivers. The spatio-temporal analysis of separate parameters did not help defining the trophic status of rivers. The assumption that correlations exist between certain parameters, guides us to use the Principal Component Analysis (PCA) in spite of conventional methods. This statistical tool was used to define the pollution levels in rivers. It also leads to observe the impact of pollutants inflows on different sites studied. To follow the fate of fecal coliform in water, the ACP of microbiological variables showed the persistence of bacterial colonies in water despite the various climatic conditions and the Mediterranean flow rate. Indeed, sediments are potential reservoirs of pathogenic microorganisms. The bacteria aggregated to the sediment can be present again in water column causing an alarming public health problem.

Des supramolécules basées sur la reconnaissance de viologènes par des curcubit[n]urils (CB[n]) ont été étudiées. Les systèmes développées incluent des [3]-, [4]- et [7]pseudorotaxanes, composés de multimères à base de viologène et de CB[7] ou CB[8]. L’étude physicochimique a montré que ces supramolécules sont interconverties électrochimiquement entre un état complexé, défini par les pseudorotaxanes, et un état dissocié comprenant chacun des partenaires. Le désassemblage résulte d’une pimérisation intra/intermoléculaire entre radicaux viologènes.L’étude physicochimique de complexes pentacoordinés basés sur la reconnaissance d’azo-aryl-imidazoles par une porphyrine à anse phénanthroline a été menée; l’objectif étant d’utiliser les propriétés d’isomérisation photoinduite trans-cis d’azo-chromophores pour dissocier ces complexes. Les complexes initiaux sont alors régénérés par relaxation thermique. La stabilité et les propriétés cinétiques des complexes pentacoordinés ont été évaluées. / Supramolecules based on the recognition of redox-active bipyridiniums by cucurbit[n]uril (CB[n]) have been studied. The investigated systems include a [3]-, a [4]- and a [7]pseudorotaxane, each of them composed of a multimeric viologen-based thread molecule and CB[7] or CB[8]. The physicochemical approach emphasized that these systems can be electrochemically switched between a complexed state, defined by the pseudorotaxanes, and an uncomplexed state comprising their components. The disassembly results from intra/intermolecular pimerization of the viologen radicals.A physicochemical approach of pentacoordinated complexes of a phen-strapped porphyrin with azo-arylimidazoles has been undertaken with the aim to use the photoinduced trans-cis isomerization properties of azo-chromophores to dissociate the complexes. Thermal equilibration reinstates the thermodynamically favoured complexes. The strength and the kinetic properties of these pentacoordinated species have been evaluated.

Dans le cadre du dragage de Port-Camargue (projet national ECODREDGE-MED), l'objectif principal des travaux de thèse a été d'étudier la re-sédimentation des sédiments ré-immergés et d'appréhender les transferts de matière (particulaires et moléculaires) associés. Une approche expérimentale de caractérisation a été réalisée pour appréhender ces phénomènes, aux échelles macroscopiques (mécanismes de sédimentation), mésoscopiques (transferts de matière particulaires) et moléculaires (phénomènes de sorption). Les travaux ont porté sur l'étude de cinq sédiments et de leurs fractions granulométriques. Une méthode de séparation granulaire a donc été développée et optimisée en ce sens, soulignant l'importance de vérifier la qualité de la séparation lorsque les développements portent sur les fractions granulaires. L'approche morpho-granulaire utilisée dans la caractérisation de ces systèmes a permis, dans un premier temps, de mettre en évidence le comportement cohésif des sédiments et d'identifier différents types d'agglomérats. Elle a ensuite été utilisée pour proposer une méthode de classification simple et rapide des sédiments, basée essentiellement sur la détermination, par granulométrie laser, du rapport volumique limon/sable.En second lieu, le comportement à la re-sédimentation a été appréhendé en étudiant la stabilité physico-chimique des particules dans la colonne de sédimentation grâce à un analyseur de suspensions concentrées, le Turbiscan MA2000. Cette stratégie apporte de nouvelles connaissances, notamment sur les mécanismes de sédimentation et l'impact de certains paramètres comme le rapport limon/sable, la salinité ou la fraction volumique sur ces mécanismes. Combinée à l'analyse du Carbone Organique Total (COT), des transferts de matière ont été identifiés dans la colonne de sédimentation, via les phénomènes interparticulaires (agglomération/dispersion) et les phénomènes de sorption (adsorption/désorption). / In the framework of Port-Camargue dredge (ECODREDGE-MED project), the main objective was to study the dredged sediment re-sedimentation and matter transfers (particular and molecular matter). An experimental characterization approach was realized in order to study these phenomena working at different scales : macroscopic (sedimentation mechanisms), mesoscopic (particular matter mobilization) and molecular (sorption phenomena). These research works focused on study of five sediments and their granular fractions. A dispersion granular method was developed and highlighted the importance to verify dispersion quality of processes concerning study of granular fractions. Morpho-granular approach used in theses works permitted to highlight cohesive aspect of sediments and to identify different agglomerate types. This approach was also used to propose a classification method of sediments, based on limon/sand ratio determination by laser granulometry.Secondly, re-sedimentation behavior was investigated studying physico-chemical stability of particles in the water column with a suspension analyszer (Turbiscan MA2000). The results contributed knowledge on sedimentation mechanisms and parameter (limon/sand ratio, salinity and volume fraction) influence. With the use of TOC analyze (Total Organic Carbone), matter transfers were identified in water column, via interparticular phenomena (agglomeration/dispersion) and sorption phenomena (adsorption/désorption).

L'objectif est de mieux connaître les diverses propriétés (physico-mécaniques, biologiques et chimiques) du bois de Mûrier blanc (Morus alba L.), en tant que matériau éminemment utilisé dans la fabrication de luths Iraniens. La différence de caractéristiques vibratoires de cette espèce a été mesurée en fonction de la présence d'extractibles, mais aussi pour trois traitements artisanaux qui ont été adaptés à l'échelle du laboratoire.Le bois Mûrier blanc est caractérisé par un module spécifique modéré, un amortissement plus bas que prévu et une anisotropie faible entre ses trois axes. Il est précisé que cette espèce ne peut pas être classifiée avec les mêmes normes que les autres bois utilisés pour les instruments classiques Européens. En utilisant les deux extractions indépendantes et successives, il est révélé que l'amortissement est régi par deux types d'extraits, certains l'abaissent et certains l'augmentent. Ces composés ne sont pas tous extractibles par les mêmes solvants. L'immersion de longue durée dans une eau à température ambiante ne modifie pas les propriétés mécaniques de l'espèce, toutefois, elle peut laisser le bois avec des défauts irréversibles. Un court traitement à l'eau chaude (70°C) entraîne l'augmentation de l'amortissement et la diminution du module spécifique. Dans ce dernier cas, des modifications sont presque progressives avec le temps.Le bois de Mûrier blanc semble être « très durable » par rapport à une attaque fongique, même dans des conditions extrêmes. Le délavage dans l'eau n'a pas d'effets sur la résistance de cette espèce contre basidiomycètes. Ce bois est classé « moyennement durable » contre l'attaque des termites et devient sensible après le délavage. Ses extractibles ont donc des effets toxique sur les termites. Les phénols, acides gras, stérols, hydrocarbures supérieurs, et composés aromatiques sont les composés présents dans les extraits de cette espèce. Le résorcinol, composé prédominant trouvé dans l'espèce, a été récemment rapporté comme la cause de plusieurs problèmes de santé, ce qui est aujourd'hui observé chez les artisans Iraniens. / This work aims at studying the several divers properties (Physico-mechanical, biological and chemical) of white Mulberry (Morus alba L.), as the leading material used in fabrication of Iranian lutes. The vibrational characteristic of this species is measured in relation with secondary metabolites (extractives) as well as three artisanal hygro-thermal treatments adapted to laboratory scale.White Mulberry wood is characterized with a moderate specific modulus, less than expected damping and a low anisotropy between three directions. It is specified that this species cannot be described with the same standards as other woods used in European classical instruments. Using independent and successive extractions reveal that two types of extraneous compounds govern the damping in this wood, some raising it, when the rest have a decreasing effect. These compounds are not all extractable by the same solvents.Long time immersion in water at ambient temperature hardly changes mechanical properties of the species, however, it can leave wood with irreversible defects. Hot water treatment at 70°C results for damping and moduli to be increased and decreased respectively. These changes are time related. Several mild desorption and adsorption cycles, seem to reduce damping without greatly endangering the moduli. White Mulberry wood is found to be very durable towards fungi, even in extreme conditions. Water leaching seems to be affectless on this species resistance towards basidiomycetes. This wood is rated moderately durable towards termites and becomes sensible after water leaching. Extractives seem to play important role in this wood natural resistance, as they are found to have toxic effect on termites. Resorcinol, a phenol, is the leading compound in the extractives of white Mulberry. Fatty acids, sterols, higher hydrocarbons, and aromatic compounds are also found as the constituents of secondary metabolites. Resorcinol is documented to cause irritations close to what is normally experienced by the fabricants working with white Mulberry for a long time.

La pyrogazéification est un procédé de conversion thermochimique prometteur pour la valorisation énergétique des biomasses et des déchets. Ce procédé conduit à la production d’un vecteur énergétique gazeux appelé « syngas » composé principalement de CO et d’H2 mais contenant également de nombreux polluants issus des déchets entrants et/ou générés au cours de la conversion. En fonction de sa pureté, le syngas peut être valorisé dans de nombreuses applications. Cependant, la formation simultanée de résidus solides (chars) sans voie de valorisation, ainsi que le coût élevé de l'étape de purification du syngas freinent le développement industriel de cette filière. Cette thèse s'intéresse à ces deux problématiques en étudiant la valorisation des chars de pyrolyse, avec ou sans adjonction de fonctions chimiques, comme adsorbants et catalyseurs pour la purification du syngas. Dans cette étude, les chars ont été produits par pyrolyse de déchets générés sur des navires de croisière et générés en quantités importantes par les sociétés modernes : Bois de Palettes Usagées, Boues de Coagulation-Floculation et Déchets Alimentaires. Une séquence de caractérisations multi-échelle a été mise en place afin de relier les caractéristiques physico-chimiques des chars aux conditions de production ainsi qu’à la nature des déchets entrants. Les chars résultants du mélange BF/DA montrent une composition chimique riche en espèces minérales tandis que les chars produits à partir de BPU sont des matériaux très majoritairement composés de carbone. L’activation à la vapeur ne modifie pas significativement la composition des chars, mais permet de développer efficacement leur porosité. L’étude s’est ensuite intéressée aux relations existant entre les propriétés physico-chimiques des chars et leur efficacité épuratoire. La capacité d’épuration d’H2S des matériaux s’est trouvée significativement améliorée par des surfaces spécifiques élevées, de hautes teneurs en espèces minérales et un pH de surface basique. Les propriétés les plus influentes pour l’activité catalytique des chars pour le craquage des goudrons (l’éthylbenzène et le benzène sont pris comme références) en gaz légers sont : la présence d’espèces minérales, la porosité et la présence de structures carbonées désordonnées dans la matrice du char. / The pyrogasification is a thermochemical process that consists in converting biomass and/or waste into a gaseous energy carrier named syngas. This syngas is mainly composed of H2 and CO but also contains many pollutants (such as tars, H2S, HCl, particles…) that must be removed before further utilization of the syngas (electricity and heat production, synthesis of biofuel or chemicals…). The production of solid residues (chars) and the cost of the syngas purification process jeopardize the industrial development of this process. This thesis aims at studying the in-situ valorisation of the pyrolysis chars, functionalized or not, as sorbent or catalyst for the syngas cleaning. In this study, pyrolysis chars have been produced by the pyrolysis of wastes generated on cruise-ships: Used Wood Pallets (UWP), Coagulation-Flocculation Sludge (CFS) and Food Waste (FW). A set of multi-scale characterizations has been performed in order to identify relationships between the physico-chemical properties of the chars, the production conditions and the nature of the initial biomass. Chars from the mixture of FW/CFS have high mineral contents while chars from UWP are mainly carbonaceous materials. The steam activation only slightly modifies the chemical composition of the chars but significantly increases their porosity. Then, the study focused on the relationships between the physico-chemical properties of the chars and their purification efficiency. The H2S sorption capacity was strongly improved by high surface areas, large mineral contents and alkaline pH surfaces. The most important properties for the catalytic activity of the chars for tar cracking reactions were: high mineral contents, large surface areas and the presence of disorganized carbon structures in the char.

Increasing socio-economic growth and development of South Africa’s freshwater systems require continuous augmentation of water sources to meet the growing water requirements of communities and industries. Anthropogenic disturbances have caused the water quality of many freshwater systems to drastically deteriorate due to constant disposal of domestic, industrial, and agricultural waste into surface waters. Government agencies make use of biomonitoring programmes to effectively manage the countries’ freshwater resources. These programmes use a variety of biological indicators (e.g., macroinvertebrates, fish, diatoms and algal species) and physico-chemical variables to determine the state of the environment. However, attempts to use microbial community structures as bioindicators of anthropogenic perturbations are greatly neglected. This study used molecular techniques (PCR-DGGE and 454-pyrosequencing) and multivariate analysis to develop a robust monitoring technique to determine the impacts of environmental disturbances on bacterial community compositions in river systems in the North West Province. Significant contributions made by this project included the establishment of a bacterial diversity framework for South African freshwater systems that are impacted by a variety of anthropogenic activities (e.g., urban and informal settlements, agriculture and mining). Furthermore, case studies demonstrated the prevalence of specific taxa at polluted sites, as well as positive and negative associations between taxa and environmental variables and pollutants. Finally, biogeochemical cycles could be partially matched to bacterial community structures in river systems. The first part of the project included a pilot study that investigated bacterial structures in a segment of the Vaal River in response to environmental parameters using molecular techniques and multivariate analysis. The most important observations made during this study included the generation of a larger bacterial diversity dataset by pyrosequencing compared to PCR-DGGE. In addition, metagenomic and multivariate analyses provided clues about potential biogeochemical roles of different taxa. The second and third part of the project included two case studies that investigated bacterial communities in the Mooi River and Wonderfonteinspruit in response to environmental activities. Both these systems are impacted by a variety of external sources such as urban and informal settlements, agriculture, and mining. The results demonstrated that perturbations nearby the Mooi River and Wonderfonteinspruit caused the overall water quality to deteriorate which in turn had a profound impact on bacterial community composition. Bacterial community structures at reference/control sites (Muiskraal and Turffontein dolomitic eye) had overall high species diversity (richness and evenness), whereas polluted sites showed lower species diversity and were dominated by the Beta- and Gammaproteobacteria, Bacteroidetes, and Verrucomicrobia. In addition, various potential pathogens (e.g. Eschirichia/Shigella, Legionella, Staphylococcus, Streptococcus etc.) were identified at impacted sites. Multivariate analysis suggested that bacterial communities and certain taxa (Malikia, Algoriphagus, Rhodobacter, Brevundimonas and Sphingopyxis) at polluted sites were mainly impacted by temperature, pH, nutrient levels, and heavy metals. Finally, the proportion of nitrogen and sulphur bacteria corresponded well with the nitrogen and sulphur levels measured in the Wonderfonteinspruit. Based on these results, it was concluded that bacterial community structures might provide a good indicator of anthropogenic disturbances in freshwater systems and may be incorporated into biomonitoring programs. / PhD (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

The main aim of the study was to analyse the impact of physico-chemical parameters on drinking water quality, biofilm formation and antibiotic resistant bacteria in the drinking water distribution system in Mafikeng, North West Province, South Africa. Another objective was to isolate and characterise Pseudomonas and Aeromonas species from drinking water distribution system and detect the virulence gene determinants in the isolates by PCR analysis. The physico-chemical data obtained were subjected to statistical analysis using Excel 2007 (Microsoft) and SPSS (version 14.0) programmes. Pearson’s correlation product of the moment was used to determine the correlation between EC, TDS, pH and temperature. The two tailed test of significance (p<0.05) was used in order to determine the significance of the result. Antibiotic susceptibility tests were performed using Kirby-Bauer disk diffusion method. Cluster analysis based on the antibiotic inhibition zone diameter data of different organisms isolated from different sites was determined and was expressed as dendograms using Wards algorithm and Euclidean distance of Statistica version 7. Specific PCR was used to determine the identities of presumptive Pseudomonas and Aeromonas species through amplification of the gyrB, toxA and the ecfX gene fragments. Virulence gene determinants for the confirmed Pseudomonas and Aeromonas species were detected by amplifying the exoA, exoS and exoT genes and the aerA and hylH gene fragments, respectively. A Gene Genius Bio imaging system (Syngene, Synoptics; UK) was used to capture the image using GeneSnap (version 3.07.01) software (Syngene, Synoptics; UK) to determine the relative size of amplicons. Physico-chemical parameters were monitored from three drinking water sources three times a week and bacteriological quality was monitored weekly for four months from raw and treated drinking water. Water samples were analysed for pH, temperature, total dissolved solids (TDS) and electric conductivity (EC). Bacterial consortia from drinking water samples were isolated using selective media and enumerated. The results revealed a good chemical quality of water. However, the microbial quality of the water is not acceptable for human consumption due to the presence of Pseudomonas, Aeromonas, faecal coliforms (FC), total coliforms (TC) and Heterotrophic bacteria. The results showed that the drinking water is slightly alkaline with pH value ranging between7.7 to 8.32. What is of concern was the microbial quality of the water. Pseudomonas sp., faecal coliforms (FC), total coliforms (TC) and heterotrophic bacteria were present in some of the treated water samples. The most significant finding of this study is that all drinking water samples were positive for Pseudomonas sp.(>100/100ml), but also that when one considers the TDS it demonstrates that water from the Modimola Dam has an impact on the quality of the mixed water. The prevalence and antibiotic resistance profiles of planktonic and biofilm bacteria isolated from drinking water were determined. The susceptibility of these isolates was tested against 11 antibiotics of clinical interest and the multiple antibiotic resistance (MAR) patterns were compiled. The most prevalent antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. All isolates from all samples were susceptible to ciprofloxacin. However, all faecal coliforms and Pseudomonas spp. were susceptible to neomycin and streptomycin. On the contrary all organisms tested were resistant to erythromycin (100%) trimethoprim and amoxycillin. Cluster analysis based on inhibition zone diameter data could not differentiate the various isolated into sample types. The highest prevalence of antibiotic resistant isolates was observed in Modimola Dam and Molopo eye. Biofilms were investigated in both raw water and treated drinking water sources for the presence of faecal coliforms, total coliforms, Pseudomonas spp., Aeromonas spp. and heterotrophic bacteria based on conventional microbiology and molecular methods. Drinking water biofilms were grown twice and the biofilm developing device containing copper and galvanized steel coupons were utilized. The Mini Tap filter, a home water treatment device which can be used at a single faucet, under constant flow was used during the second collection of treated water samples from cold water taps. Scanning electron micrograph revealed the existence of biofilms in all the sites investigated and the highest density was obtained on galvanized steel coupons. Isolates were tested against the antibiotics ampicillin (10μg), cephalothin (5μg), streptomycin (10μg), erythromycin (15μg), chloramphenicol (30μg), neomycin (30 μg), amoxycillin (10 μg), ciprofloxacin (5 μg), trimethoprim (25μg), kanamycin (30μg), and oxytetracycline (30μg). The multiple antibiotic resistance profiles and the presence of virulence related genes were determined. Various types of drug resistance and presence of virulence genes were observed. The most prevalent resistance phenotype observed was KF-AP-C-E-OT-TM-A. In conclusion, the results indicated the occurrence of faecal indicator bacteria in the drinking water destined for human consumption. Faecal indicator bacteria are the major contributors of poor drinking water quality and may harbour opportunistic pathogens. This highlighted survival of organisms to treatment procedures and the possible regrowth as biofilms in plumbing materials. The detection of large proportion of MAR Aeromonas and Pseudomonas species which possessed virulent genes was a cause of concern as these could pose health risks to humans. The data obtained herein may be useful in assessing the health risks associated with the consumption of contaminated water. / PhD (Microbiology), North-West University, Potchefstroom Campus, 2014

Increasing socio-economic growth and development of South Africa’s freshwater systems require continuous augmentation of water sources to meet the growing water requirements of communities and industries. Anthropogenic disturbances have caused the water quality of many freshwater systems to drastically deteriorate due to constant disposal of domestic, industrial, and agricultural waste into surface waters. Government agencies make use of biomonitoring programmes to effectively manage the countries’ freshwater resources. These programmes use a variety of biological indicators (e.g., macroinvertebrates, fish, diatoms and algal species) and physico-chemical variables to determine the state of the environment. However, attempts to use microbial community structures as bioindicators of anthropogenic perturbations are greatly neglected. This study used molecular techniques (PCR-DGGE and 454-pyrosequencing) and multivariate analysis to develop a robust monitoring technique to determine the impacts of environmental disturbances on bacterial community compositions in river systems in the North West Province. Significant contributions made by this project included the establishment of a bacterial diversity framework for South African freshwater systems that are impacted by a variety of anthropogenic activities (e.g., urban and informal settlements, agriculture and mining). Furthermore, case studies demonstrated the prevalence of specific taxa at polluted sites, as well as positive and negative associations between taxa and environmental variables and pollutants. Finally, biogeochemical cycles could be partially matched to bacterial community structures in river systems. The first part of the project included a pilot study that investigated bacterial structures in a segment of the Vaal River in response to environmental parameters using molecular techniques and multivariate analysis. The most important observations made during this study included the generation of a larger bacterial diversity dataset by pyrosequencing compared to PCR-DGGE. In addition, metagenomic and multivariate analyses provided clues about potential biogeochemical roles of different taxa. The second and third part of the project included two case studies that investigated bacterial communities in the Mooi River and Wonderfonteinspruit in response to environmental activities. Both these systems are impacted by a variety of external sources such as urban and informal settlements, agriculture, and mining. The results demonstrated that perturbations nearby the Mooi River and Wonderfonteinspruit caused the overall water quality to deteriorate which in turn had a profound impact on bacterial community composition. Bacterial community structures at reference/control sites (Muiskraal and Turffontein dolomitic eye) had overall high species diversity (richness and evenness), whereas polluted sites showed lower species diversity and were dominated by the Beta- and Gammaproteobacteria, Bacteroidetes, and Verrucomicrobia. In addition, various potential pathogens (e.g. Eschirichia/Shigella, Legionella, Staphylococcus, Streptococcus etc.) were identified at impacted sites. Multivariate analysis suggested that bacterial communities and certain taxa (Malikia, Algoriphagus, Rhodobacter, Brevundimonas and Sphingopyxis) at polluted sites were mainly impacted by temperature, pH, nutrient levels, and heavy metals. Finally, the proportion of nitrogen and sulphur bacteria corresponded well with the nitrogen and sulphur levels measured in the Wonderfonteinspruit. Based on these results, it was concluded that bacterial community structures might provide a good indicator of anthropogenic disturbances in freshwater systems and may be incorporated into biomonitoring programs. / PhD (Environmental Sciences), North-West University, Potchefstroom Campus, 2015

The main aim of the study was to analyse the impact of physico-chemical parameters on drinking water quality, biofilm formation and antibiotic resistant bacteria in the drinking water distribution system in Mafikeng, North West Province, South Africa. Another objective was to isolate and characterise Pseudomonas and Aeromonas species from drinking water distribution system and detect the virulence gene determinants in the isolates by PCR analysis. The physico-chemical data obtained were subjected to statistical analysis using Excel 2007 (Microsoft) and SPSS (version 14.0) programmes. Pearson’s correlation product of the moment was used to determine the correlation between EC, TDS, pH and temperature. The two tailed test of significance (p<0.05) was used in order to determine the significance of the result. Antibiotic susceptibility tests were performed using Kirby-Bauer disk diffusion method. Cluster analysis based on the antibiotic inhibition zone diameter data of different organisms isolated from different sites was determined and was expressed as dendograms using Wards algorithm and Euclidean distance of Statistica version 7. Specific PCR was used to determine the identities of presumptive Pseudomonas and Aeromonas species through amplification of the gyrB, toxA and the ecfX gene fragments. Virulence gene determinants for the confirmed Pseudomonas and Aeromonas species were detected by amplifying the exoA, exoS and exoT genes and the aerA and hylH gene fragments, respectively. A Gene Genius Bio imaging system (Syngene, Synoptics; UK) was used to capture the image using GeneSnap (version 3.07.01) software (Syngene, Synoptics; UK) to determine the relative size of amplicons. Physico-chemical parameters were monitored from three drinking water sources three times a week and bacteriological quality was monitored weekly for four months from raw and treated drinking water. Water samples were analysed for pH, temperature, total dissolved solids (TDS) and electric conductivity (EC). Bacterial consortia from drinking water samples were isolated using selective media and enumerated. The results revealed a good chemical quality of water. However, the microbial quality of the water is not acceptable for human consumption due to the presence of Pseudomonas, Aeromonas, faecal coliforms (FC), total coliforms (TC) and Heterotrophic bacteria. The results showed that the drinking water is slightly alkaline with pH value ranging between7.7 to 8.32. What is of concern was the microbial quality of the water. Pseudomonas sp., faecal coliforms (FC), total coliforms (TC) and heterotrophic bacteria were present in some of the treated water samples. The most significant finding of this study is that all drinking water samples were positive for Pseudomonas sp.(>100/100ml), but also that when one considers the TDS it demonstrates that water from the Modimola Dam has an impact on the quality of the mixed water. The prevalence and antibiotic resistance profiles of planktonic and biofilm bacteria isolated from drinking water were determined. The susceptibility of these isolates was tested against 11 antibiotics of clinical interest and the multiple antibiotic resistance (MAR) patterns were compiled. The most prevalent antibiotic resistance phenotype observed was KF-AP-C-E-OT-K-TM-A. All isolates from all samples were susceptible to ciprofloxacin. However, all faecal coliforms and Pseudomonas spp. were susceptible to neomycin and streptomycin. On the contrary all organisms tested were resistant to erythromycin (100%) trimethoprim and amoxycillin. Cluster analysis based on inhibition zone diameter data could not differentiate the various isolated into sample types. The highest prevalence of antibiotic resistant isolates was observed in Modimola Dam and Molopo eye. Biofilms were investigated in both raw water and treated drinking water sources for the presence of faecal coliforms, total coliforms, Pseudomonas spp., Aeromonas spp. and heterotrophic bacteria based on conventional microbiology and molecular methods. Drinking water biofilms were grown twice and the biofilm developing device containing copper and galvanized steel coupons were utilized. The Mini Tap filter, a home water treatment device which can be used at a single faucet, under constant flow was used during the second collection of treated water samples from cold water taps. Scanning electron micrograph revealed the existence of biofilms in all the sites investigated and the highest density was obtained on galvanized steel coupons. Isolates were tested against the antibiotics ampicillin (10μg), cephalothin (5μg), streptomycin (10μg), erythromycin (15μg), chloramphenicol (30μg), neomycin (30 μg), amoxycillin (10 μg), ciprofloxacin (5 μg), trimethoprim (25μg), kanamycin (30μg), and oxytetracycline (30μg). The multiple antibiotic resistance profiles and the presence of virulence related genes were determined. Various types of drug resistance and presence of virulence genes were observed. The most prevalent resistance phenotype observed was KF-AP-C-E-OT-TM-A. In conclusion, the results indicated the occurrence of faecal indicator bacteria in the drinking water destined for human consumption. Faecal indicator bacteria are the major contributors of poor drinking water quality and may harbour opportunistic pathogens. This highlighted survival of organisms to treatment procedures and the possible regrowth as biofilms in plumbing materials. The detection of large proportion of MAR Aeromonas and Pseudomonas species which possessed virulent genes was a cause of concern as these could pose health risks to humans. The data obtained herein may be useful in assessing the health risks associated with the consumption of contaminated water. / PhD (Microbiology), North-West University, Potchefstroom Campus, 2014