Mawira Sitima is a thermal spring located in the village Sitima outside of Liwonde,Malawi. The spring is used by many of the local villagers to take baths, play around and wash clothes. Washing using detergents in springs may cause harm to the aquatic ecosystems and worsen the water quality, which might pose a health risk to people swimming in the spring. This study compared physico-chemical properties of Mawira Sitima and the soil and plants around it with standards, guidelines and other studies to assess thewater quality of the spring. The water quality of another hot spring, Mawira Liwonde, was also assessed and compared to Mawira Sitima to investigate the potential for recreationaluse of Mawira Sitima. The findings were then used to produce recommendations for future monitoring of relevant variables in the spring. The results indicate that the current water quality of Mawira Sitima is of no concern for the local health and has all basis to be of recreational use. A monitoring system including electrical conductivity, water temperature, pH, total phosphorous, copper, manganese, and iron was recommended based on correlations and trends. Monitoring of extreme meteorological conditions, such as heavy rainfall or drought, were also recommended. / Mawira Sitima är en varm källa belägen i byn Sitima utanför Liwonde, Malawi. Källan används av lokalbefolkningen för att bada, leka och tvätta kläder. Att tvätta med tvättmedel i källor kan skada de akvatiska ekosystemen och försämra vattenkvaliteten vilket kan leda till hälsorisker för de som använder källan. I denna studie jämfördes fysikaliska och kemiska parametrar i Mawira Sitima, och jorden och plantor runtom den, med standarder, riktlinjer och andra studier för att utvärdera vattenkvaliteten i källan. Vattenkvaliteten i en annan källa, Mawira Liwonde, utvärderades också och jämfördes med Mawira Sitima i syftet att undersöka potenitalen för fritidsbruk av Mawira Sitima. Undersökningsresultaten användes sedan för att föreslå ett framtida övervakningssystem av relevanta variabler i källan. Resultaten i studien visar på god vattenkvalitet i Mawira Sitima och stora möjligheter för fritidsbruk. Övervakningssystemet föreslogs innefatta elektrisk konduktivitet, vattentemperatur, pH, totalfosfor, koppar, mangan och järn baserat på korrelationer och trender. Även extremväder såsom stora mängder regn eller extrem torka rekommenderades ingå i övervakningssytemet

Southeast Harney Lake Hot Spring and its effluent channel were examined for spatial and temporal variations in the concentrations of several chemical species, viz. dissolved oxygen, pH, alkalinity, phosphate-phosphorous, nitrate and nitrite-nitrogen, conductivity, chloride, flouride, temperature, calcium, magnesium, potassium, and sodium. Phosphate-phosphorous and nitrate-nitrogen both exhibited downstream increases in concentration. pH and alkalinity increased also, mainly due to CO2 evolution and temperature mediated carbon species redistribution. The diurnal pH and alkalinity curves exhibited depressions during the daylight hours due to the oxidation of ammonia to nitrate within the pool and effluent stream.

M. Tech. (Department of Biotechnology, Faculty of Applied and Computer Sciences), Vaal University of Technology. / The use of conventional culture-based approach results in vast majority of microbes (90 - 99%) unaccounted for. However, over the past years, the use of metagenomics, which is a culture-independent comprehensive approach has enabled researchers to access nearly 100% of the microbiome. In this study, three hot springs (44 – 70 oC) in Limpopo province of South Africa were investigated as potential sources of genes encoding for DNA-manipulating enzymes (DNA polymerase, DNA ligase and endonuclease), which are central in genetic engineering. They are usually grouped into four broad classes (nucleases, ligases, polymerases and modifying enzymes) depending on the type of the reaction they catalyze. Accordingly, hot spring metagenomic DNA was successfully extracted using modified SDS-CTAB method involving gel purification and electroelution. Consequently, a portion of the extracted metagenomic DNA was used for sequencing and another for fosmid library construction. Sequencing was done using Illumina MiSeq next generation sequencing platform and sequence data analyzed and de novo-assembled using CLC Genomic Workbench, which resulted in 5 681 662 reads and 7 338 contigs. A metagenome expression fosmid library of approximately 2.16 x 103 clones was also constructed using CopyControl™ HTP Fosmid Library Production Kit with pCC2FOS™ Vector. A BLAST algorithm in NCBI revealed 57 distinct genes for DNA polymerase, 29 genes for DNA ligase and more than 100 genes for endonuclease II enzymes. Hence, three genes related to thermophiles representing genes for DNA polymerase, DNA ligase and endonuclease II were selected. Accordingly, the three genes were codon-optimized, synthesized and successfully cloned into pET- 30a (+) and overexpressed in Escherichia coli BL21 (DE3) by inducing with 0.5 mM IPTG and incubating overnight at 16ºC. The cells were lysed using B-PER Reagent, protein extracted and purified using AKTA start protein purification system and purity of 85- 95 % was achieved. From this study, it can be concluded that metagenomics as an approach, can be used to mine for putative DNA-manipulating enzymes from hot spring metagenome. Besides, further study should be conducted to formulate the developed DNA-manipulating enzymes and study the practical application and chart way for commercialization. Moreover, the constructed fosmid library could also be screened for potentially novel thermo-stable biomolecules of industrial and therapeutic importance.

Les virus peuvent être retrouvés dans tous les écosystèmes où de la vie est présente. Ils constituent l’entité biologique la plus abondante de la biosphère. Si de nombreuses données sont disponibles sur l’abondance et la dynamique virale dans les écosystèmes aquatiques tempérés, peu d’études ont été menées sur ces aspects dans les milieux extrêmes, dont les sources hydrothermales. Dans l’étude présentée dans ce manuscrit, les communautés procaryotiques et virales des sources hydrothermales des Terres australes et antarctiques françaises (TAAF) ont été explorées. Dans un premier temps, les cellules procaryotiques et les particules de type viral (VLP) ont été dénombrées dans plusieurs sources chaudes terrestres et marines côtières. L’abondance microbienne et virale est de l’ordre de 105 - 106 particules/ml dans les deux types de sources avec des rapports VLP/procaryotes (VPR) qui sont généralement faibles, concordant ainsi avec rares les données disponibles actuellement dans la littérature. Dans un second temps, la diversité morphologique des VLP a été analysée par observation au microscope électronique à transmission. La présence de VLP de morphologies différentes a pu être constatée dans quelques échantillons bruts, mais également dans des cultures d’enrichissement, où elles étaient associées à des Thermococcales et des Thermotogales. Finalement, quelques souches isolées de ces échantillons ont été criblées pour la présence de virus aboutissant à la description d’un nouveau bactériovirus tempéré associé à une bactérie thermophile Geobacillus. L’effet d’un choc osmotique en présence de NaCl et l’effet d’un stress anoxique sur la production virale ont également été étudiés. La caractérisation du virus GTV1 a ensuite été entamée. Il appartient à la famille des Myoviridae et a un génome composé d’ADN double brin de 38841 pb, composé de 71 ORF prédits. Enfin, l’étude de la diversité microbienne a permis de décrire une nouvelle espèce bactérienne hautement thermophile, Calditerricola clavaformis sp.nov. / Viruses thrive in all types of ecosystems where life is found. They represent the most abundant biological entity of our biosphere. Though several studies have been conducted on viral abundance and dynamics in mesophilic aquatic ecosystems, these aspects remain largely unexplored in extremophilic environments, such as hot springs. In this study, prokaryotic and associated viral communities of the French Southern and Antarctic Lands hot springs were explored. First, prokaryotic cells and Virus-like particles (VLP) were enumerated in several terrestrial and inshore hot springs. The results reveal an abundance of 105 - 106 particles/ml in both types of hot springs studied. The virus-to-prokaryote ratios (VPR) were generally low, confirming thus actual knowledge in these types of ecosystems. The morphological diversity of VLP was then studied in raw samples as well as in enrichment cultures containing Thermococcales and Thermotogales. Several isolates obtained from these samples were then screened for viral particles which led to the discovery and description of a temperate phage (GTV1) of a thermophilic bacterium belonging to the genus Geobacillus. The effect of NaCl and anoxic stress on the viral production was studied. The genomic characterization of the GTV1 was started and revealed a 38441 bp genome with 71 predicted ORF. Finally, microbial diversity studies led also to the discovery of a new extremely thermophilic bacterium, Calditerricola clavaformis sp.nov.

Thermal waters in Karlovy Vary are characterized by carbonate sedimentation in places of their emergence. These sediments contain radium. Radium bond in every type of sinter (aragonite, calcite) has been investigated experimentally by acid sinter dissolution. It has been found out, that radium can join dissoluble fraction in both sinter types . The original hypothesis that radium can bond only aragonite-type of sinter has been refuted. Additional experiments have been processed using synthetic radiobarite. Present radium in radiobarite can be partly dissoluted using acids. If radium was incorporated in radiobarite (it is known from locality Vřídlo in Karlovy Vary and elsewhere) then maybe it would be partly dissoluted in experiments.

Le travertin, connu sous le terme de lapis tiburtinus dans l’Antiquité romaine, est une roche issue de la précipitation du carbonate dissous dans les eaux de sources chaudes (travertins) ou froides (tufs calcaires), sous l’influence de processus physico-chimiques et/ou biologiques. Ce phénomène est décrit dès l’époque gréco-romaine par les auteurs antiques (Strabon, Pline l’Ancien, Vitruve), qui témoignent d’une roche qui se forme sous leurs yeux, qui dessine le paysage et qui est largement exploitée pour la construction (p. ex. le Colisée à Rome, le Temple grec de Ségeste en Sicile). Abondamment répartis à la surface de la Terre et caractérisés par une certaine diversité morphologique, les travertins représentent d’excellents enregistreurs des conditions climatiques et hydrologiques de leur dépôt, offrant un potentiel très fiable d’archives sédimentaires utilisées au sein de problématiques paléoenvironnementales. La notion de travertins anthropiques définie ici prend en compte l’influence de l’Homme sur ces formations sédimentaires et les eaux qui leur sont associées. Elle est illustrée par une approche géoarchéologique des dépôts de travertins préservés sur le site antique de Jebel Oust, en Tunisie, où l’exploitation d’une source chaude est attestée depuis le début de notre ère jusqu’à son tarissement dès la fin de l’Antiquité tardive. La source thermale surgissant sur le versant oriental de la montagne fut l’objet d’un culte aux époques romaine puis paléochrétienne et alimentait en eau chaude, par le biais d’un aqueduc, un édifice thermal localisé en aval. Notre approche géoarchéologique met en lumière l’anthropisation du versant qui se traduit par un contrôle du fonctionnement de la source chaude et des dynamiques sédimentaires associées. En parallèle, l’analyse des travertins préservés au sein des structures antiques révèle des informations primordiales sur les conditions de déroulement du culte et sur les pratiques balnéaires (fonction des salles thermales, gestion de l’eau, phases de réfection, états d’abandon). Une vision plus générale d’une géoarchéologie des travertins anthropiques propose une nouvelle approche des problématiques liées à l’eau, en insistant sur la gestion plus ou moins complexe d’une source carbonatée, chaude ou froide, et en précisant le degré d’impact humain sur le développement des travertins. / Travertine, known as lapis tiburtinus during Roman times, are continental limestones precipitated in calcareous environments from thermal waters of hot springs (travertine) or cool waters of karstic springs (calcareous tufa). This phenomenon is well-known during Classical Antiquity and had been described by several ancient authors (Strabo, Pliny the Elder, Vitruvius) who depicted a stone that forms extremely rapidly, a stone that outlines the landscape and which is largely used for construction (e.g. The Colosseum in Roma, the Greek temple at Segesta in Sicily). These deposits are widespread on Earth’s surface showing various morphologies and are great sedimentary records of climatic and hydrologic conditions. Thus they represent valuable proxies for palaeoenvironmental studies. The notion of anthropogenic travertine takes into consideration human impact on these deposits and on travertine-depositing waters. It is documented by the study of the roman site of Jebel Oust, Tunisia, where the exploitation of a hot spring is attested from the first century A.D. to the end of Late Antiquity. The site is characterized by a temple settled around the spring’s vent associated with Roman baths located downstream and supplied with hot water via an aqueduct. Our geoarchaeological approach brings to light the anthropization of the regional geosystem expressed by an entire control over the hot spring and its associated deposits. Furthermore the study of travertines preserved in the archaeological structures reveals precious and original information about water cult and bathing practices during Antiquity (thermal rooms function, water management, repair phases, states of neglect and decay). Moreover, geoarchaeology of anthropogenic travertine intends to offer a new approach of research‘s problematic dealing with water managements and integrating human impact on travertine’s development.