This thesis focuses on developing a new method to measure trace tellurium (Te) in different environmental samples such as lake waters, mine tailings and sediments. The developed technique is based on Hydride Generation Atomic Fluorescence Spectroscopy (HG-AFS), a technique that can measure low concentration of Te and also allows for Te speciation at low cost and high efficiency in various environmental samples. To validate the method that could be used to determine Te speciation in various types of environmental samples, a series of tests has been designed for finding the best conditions to measure Te(IV) using HG-AFS and obtain accurate and reliable results. Those tests include the stability of the signal, the acidity of the solution, the volatility of Te after digestion of solids, the reduction from Te(VI) to Te(IV), the detection limit of the technique, and the validity of two digestion methods under the optimum (HG-AFS) instrumental settings. An interference study including the most common elements in the Earth’s crust such as (Ni, Fe, Pb, Cr, Cu, Co, Zn, Mn and Mo) was also performed. The results of this study showed that Cu(II) can severely interfere with Te quantification decreasing the Te signal to almost zero. Therefore, different masking agents such as 8-hydroxyquinoline, 1,10-phenanthroline, urea and thiourea were tested to reduce and eliminate this interference.

Human adenoviruses (HAdV) are non-enveloped viruses with an icosahedral capsid and a linear double-stranded DNA genome. These viruses belong to the family Adenoviridae and genus Mastadenovirus. An important property of the HAdV is that it is non-enveloped making it highly resistant to detergents and harsh environmental conditions. This virus is grouped in seven species (A-G) with more than 88 genotypes. These seven species are associated with several diseases, such as, respiratory infections, keratoconjunctivitis, urinary infections, hepatitis and gastrointestinal infections. The HAdV is one of the etiological causes of acute gastroenteritis, mainly caused by HAdV-F40 and HAdV-F41. The virus can be transmitted via the faecal-oral route, inhalation of respiratory droplets and direct contact with contaminated environments. The virus is known to be ubiquitous in environments where human contamination is likely to occur such as wastewater treatment plants. These human contaminations could occur through contaminated secretion and excretions within aqueous environments. There is currently a limited amount of information on the HAdV in water Molecular characterisation of human adenoviruses from environmental environments, particularly in Tshwane, Gauteng. Therefore, the aim of the study was to investigate the presence and genotypes of human adenovirus in environmental samples namely raw sewage and treated effluent, using molecular methods. For genotypic characterisation, Sanger sequencing was used on amplicons from 12 HAdV positive samples and next generation sequencing were used on all the amplicons from HAdV positive samples. A total of 150 environmental samples (75 raw sewage and 75 effluent) were collected from two wastewater treatment plants in Tshwane over the study period of 18 months. These environmental samples comprised of 1 L raw sewage and 10 L treated effluent samples. The primary viral recovery for the 1 L raw sewage and 10 L treated effluent samples were performed using skimmed milk flocculation procedure and glass wool adsorption elution technique, respectively. For secondary viral recovery, both environmental samples were subjected to polyethylene glycol/sodium chloride precipitation. Manual extraction was used to extract the nucleic acids from the virus concentrate with mengovirus (MV) used as an extraction control. For the quantification of HAdV, standard curves prepared from known dilutions of HAdV and MV were used. Human adenovirus was detected in 140/150 (93%) of the environmental samples comprising of 69/75 (92%) being raw sewage and 71/75 (95%) being effluent samples. The HAdV concentrations detected in wastewater treatment plant 1 (WWTP 1) ranged from 1.38x105 gc/L to 4.50 x 109 gc/L for raw sewage and 5.08x103 gc/l to 4.30x108 gc/L for effluent. The HAdV concentrations detected in WWTP 2 ranged from 6.84x104 gc/L to 1.69x1012 gc/L for raw sewage and 5.27x103 gc/L to 1.16x108 gc/L for effluent. The HAdV hexon amplification success rate from the nucleic acids was 43/140 (31%). Eighteen HAdV genotypes were successfully characterised using Sanger sequencing. The HAdV-D was the most predominant species in both WWTPs, follow by HAdV-B and HAdV-F. The HAdV-A and HAdV-E species were the least identified. Next generation sequencing identified four times as many genotypes as Sanger sequencing (77 different genotypes). The HAdV-D (types 8, 9, 13, 17, 19, 20, 23, 24, 28, 29, 32, 33, 36, 42, 44, 47, 49, 51, 56, 60, 62, 64, 67 and 81) and HAdV-B (types 2, 3, 7, 11 and 66) were the most predominant species followed by HAdV-F (types 40 and 41), HAdV-A (types 12 and 76), HAdV-E ( type 4) and HAdV-C (type 1). Testing wastewater treatment plants is advantageous as it allows for the detection and identification of HAdV types circulating in the surrounding communities. Due to the large number of species identified using NGS, it is the superior typing method and should be used for future studies. These include strains causing symptomatic and asymptomatic infections. Human adenovirus was detected at comparable frequencies in raw sewage and treated effluent wastewater, with slightly higher detection in effluent samples. However, the viability of these viruses is unknown and should be investigated in further studies. The detection of viruses in wastewater treatment plants are a public health concern as the treated effluent is discharged into rivers, which may be used by communities for domestic and recreational purposes. / Dissertation (MSc (Medical Virology))--University of Pretoria, 2020. / NRF, PRF / Medical Virology / MSc (Medical Virology) / Restricted

Metaproteomics, as a proteomic approach to analyse environmental samples, is a new and expanding field of research. The field promises new ways of determining the status of the organisms present in a sample, and could provide additional information compared to metagenomics. Being a novel field of research, robust methods and protocols have not yet been established. In this thesis, we examine several methods for a reliable extraction of protein from soil and periphyton samples. The extraction should preferably be fast, compatible with downstream analysis by mass spectrometry and extract proteins in proportion to their presence in the original sample. A variety of methods and buffers were used to extract proteins from soil and periphyton samples. Concentration determinations showed that all of these methods extracted enough protein for further analysis. For purification and digestion of the samples, several methods were used. The purified samples were analysed on three different mass spectrometers, with the Orbitrap Velos Pro delivering the best results. The results were matched against four genomic and metagenomic databases for identification of proteins, of which the UniProt/SwissProt database gave the best result. A maximum of 52 proteins were identified from periphyton samples when searching against UniProt/SwissProt with strict settings, of which the majority were highly conserved proteins. The main limitation for this type of work is currently the lack of proper metagenomic databases.

There is a need for a sensitive, specific, rapid and cost-effective assay that can be used as an early warning signal of contamination of aquatic ecosystems. The purpose of this work was to develop a sensitive stress-specific microbial bioreporter responsive to pro-oxidants. Furthermore, the bioreporter was designed to be applicable in environments possibly affected by metal processing activities. An E.coli bioreporter was developed containing a plasmid with the katG promoter sequence as the sensing sequence and with mCherry as the reporter protein. The bioreporter responded to metal pro-oxidants (Cd, As, Zn, Pb, Ag and Ag nanoparticles). A new assay growth-medium was developed and contributed to improve the sensitivity of our assay that has the best detection limit to inorganic pro-oxidants compared to other oxidative-stress sensitive bioreporters in the literature. The bioreporter detected pro-oxidants in environmental samples. The assay has a reasonable sensitivity, however, it still lacks sensitivity to detect pro-oxidants at concentrations lower than those shown to be toxic to many aquatic species. Within-lab reproducibility and robustness were determined to be acceptable. For stress-specific bioreporters to be incorporated in regulative legislations and industrial monitoring programs there is a need to improve the sensitivity of these assays, they need to be calibrated with other relevant pro-oxidants, inter-lab reproducibility needs to be established and robustness to environmental samples needs to be further tested. To further validate the sensitivity and ecotoxicological relevance of the bioreporter as a relevant predictive tool, stress-specific bioreporter assays need to be performed in parallel with traditional ecotoxicological assays using contaminated environmental samples.

No description available.

Endocrine disruptors represent a group of chemical compounds that are able to negatively influence the hormonal system of vertebrates. Environmental Protection Agency (U.S.EPA) defines these compounds as exogenous substance or mixture that interferes with the synthesis, secretion, transport, binding, activity, or degradation of natural hormones. This can be observed at the level of the individual organism, its progeny, populations and subpopulations. All these changes have negative effects on homeostasis, reproduction, development or change the behavior of the affected animals. This work focuses on 7 endocrine disruptors - natural estron, 17β estradiol, estriol, and synthetic 17α-ethynylestradiol, irgasan (triclosan), 4-nonylfenol, bisphenol A in the influent and effluent of wastewater plants in the Czech Republic. The thesis contains an optimization of endocrine disruptors determination in wastewater, a preliminary screening to determinate concentration levels, 24 hours composite samples and monitoring of one selected wastewater plant during a day. The analytical procedure is based on filtration, solid-phase extraction (SPE), gel permeation chromatography (GPC), derivatization and gas chromatography coupled with mass spectrometry (GC/MS). Keywords: endocrine disruptors, wastewaters, Czech...

A cultivation-independent approach, sequence analysis of 18S rRNA genes PCR-amplified from environmental DNA, was used to explore the diversity and distribution of eukaryotic microbes inhabiting algal mats in two acidic geothermal streams in Yellowstone National Park. The objectives were to: (1) clarify the identity of mat forming algae in Nymph Creek (2) survey microbial species in the Nymph Creek mat over seasonal intervals along a thermal gradient (3) compare microbial species in the Nymph Creek mat with those in Alluvium Creek mats (4) evaluate microbial species in algal mats formed on different substrates in Alluvium Creek. The results show that a novel red alga dominates high temperature regions (~50ºC) of Nymph Creek and two "Chlorella-like" algae predominate the cooler regions (<38ºC). The predominant algae in Alluvium Creek were distinctly different from those in Nymph Creek. Several stramenophiles and fungi were detected in each algal mat.

174f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-04-03T14:55:10Z No. of bitstreams: 1 Tese Cleber.pdf: 3023251 bytes, checksum: 3c19c67227656dc28f623ea371c6e7dd (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-04-23T15:23:58Z (GMT) No. of bitstreams: 1 Tese Cleber.pdf: 3023251 bytes, checksum: 3c19c67227656dc28f623ea371c6e7dd (MD5) / Made available in DSpace on 2013-04-23T15:23:58Z (GMT). No. of bitstreams: 1 Tese Cleber.pdf: 3023251 bytes, checksum: 3c19c67227656dc28f623ea371c6e7dd (MD5) Previous issue date: 2011 / CAPES / Nesta tese é apresentada a aplicação de técnicas quimiométricas na otimização de métodos usando a espectrometria de emissão óptica com plasma indutivamente acoplado (ICP OES) e espectrometria de absorção atômica com chama (F AAS) visando a análise de amostras ambientais. No primeiro trabalho, variáveis como potência de radiofrequência, vazão do gás do plasma, vazão do fluxo de nebulização, vazão do gás auxiliar, vazão da amostra e concentração de ácido nítrico na amostra foram otimizadas através da aplicação de planejamento fatorial completo e Box-Behnken. Parâmetros analíticos como precisão, robustez, sensibilidade e exatidão mostraram-se adequados para determinação de Al, B, Ba, Ca, Cd, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Ni, Pb, Si, Sn, Sr, Ti, Tl, V e Zn em amostras de água subterrânea, água de rio, efluente e solo usando ICP OES. A estratégia de medição e o tempo total de análise também foram otimizados. O novo método foi implantado na rotina do laboratório da CETREL e está proporcionando uma enorme redução no consumo do argônio, em torno de 35 %, gerando uma economia de aproximadamente 320 m3 de argônio anuais, além da redução nos custos associados a outros acessórios do ICP OES. No segundo trabalho, três variáveis de pré-concentração do chumbo (pH, concentração do tampão e vazão da amostra) foram otimizadas empregando planejamento fatorial completo 23. O método é baseado na sorção de Pb(II) em uma minicoluna recheada com uma resina de Amberlite XAD-4 funcionalizada com 4-(5´- bromo-2´-tiazolilazo)orcinol (Br-TAO). O método apresentou limite de quantificação de 1,7 μg L-1, faixa linear de 1,7-200 μg L-1 e fator de enriquecimento de 36, para 25,0 mL de amostra. A exatidão do método foi verificada pela determinação de chumbo em material de referência certificado de água doce NIST 1643d. O método foi aplicado na determinação de chumbo em amostras de água coletadas na cidade de Jequié-BA usando F AAS. Os teores de chumbo variaram de 4,1 a 5,7 μg L-1. As ferramentas quimiométricas utilizadas nos dois trabalhos foram muito eficazes na otimização de variáveis importantes e permitiu um estudo descritivo das variáveis e suas interações. Os dois métodos apresentaram parâmetros analíticos adequados para determinação de metais e metalóides em amostras ambientais / Salvador

Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-25T15:10:49Z No. of bitstreams: 1 Dissertação - Marla Jalene Alves.pdf: 2258783 bytes, checksum: 2a65889894741102692a4ac397f43a07 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-25T15:11:05Z (GMT) No. of bitstreams: 1 Dissertação - Marla Jalene Alves.pdf: 2258783 bytes, checksum: 2a65889894741102692a4ac397f43a07 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2016-11-25T15:11:18Z (GMT) No. of bitstreams: 1 Dissertação - Marla Jalene Alves.pdf: 2258783 bytes, checksum: 2a65889894741102692a4ac397f43a07 (MD5) / Made available in DSpace on 2016-11-25T15:11:19Z (GMT). No. of bitstreams: 1 Dissertação - Marla Jalene Alves.pdf: 2258783 bytes, checksum: 2a65889894741102692a4ac397f43a07 (MD5) Previous issue date: 2015-10-28 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The melanized fungi are widely distributed in the environment and has been commonly related in human diseases such as Chromoblastomycosis, Phaeomycosis and Eumycetoma, but in the Amazon region, which have a favorable weather for the proliferation of microorganisms, there are few studies about these fungi and little is known about their distribution in this environment and its pathogenic potential. Thus, in this study were studied melanized fungi in environmental samples from rural areas of the Amazon and the relationship of these environmental isolates with melanized fungi that cause mycoses. Environmental samples were obtained in rural areas of Iranduba and Presidente Figueiredo – Amazonas (Brazil). Also entered in the study, clinical specimens deposited in Fungal Collections from Amazonas. Two hundred thirteen samples of various environmental substrates were collected: soil, parts of living plants, decaying plant material and drinking water. Of these samples, 45 isolates were subjected to identification by sequencing the ITS region, 7 species of melanized fungi that were identified have been described in the literature as etiological agents of mycoses in humans: Cladophialophora immunda, Cladosporium sphaerospermum, Cladosporium cladosporioides, Exophiala oligosperma, Exophiala jeanselmei, Hortaea werneckii and Curvularia lunata. Most isolates were obtained from thorns. Some isolates grown at 37°C temperature. The molecular identification of clinical isolates showed similarity with Fonsecaea pedrosoi, Cladosporium sphaerospermum and Knufia epidermidis. Of these clinical specimens, only the species C. sphaerospermum was isolated from the environment. This study shows that the studied environmental substrates are potential sources of infection of melanized fungi that may cause mycoses. / Os fungos melanizados estão amplamente distribuídos no ambiente e têm sido comumente relacionados à doenças em seres humanos tais como a Cromoblastomicose, a Feomicose e o Eumicetoma, porém na região Amazônica, que possui clima favorável para a proliferação de microrganismos, há raros estudos realizados sobre estes fungos e pouco se sabe a respeito da sua distribuição neste ambiente e do seu potencial patogênico. Desta forma, na presente pesquisa foram estudados fungos melanizados em amostras ambientais de áreas rurais do Amazonas e a relação desses isolados ambientais com os fungos melanizados causadores de micoses. As amostras ambientais foram obtidas em áreas rurais dos municípios de Iranduba e Presidente Figueiredo, no Amazonas (AM). Também entraram no estudo amostras clínicas depositadas em Coleções Fúngicas do AM. Foram coletadas 213 amostras de diferentes substratos ambientais: solo, partes de plantas vivas, material vegetal em decomposição e água de consumo humano. Dessas amostras, 58 isolados foram submetidos a identificação por sequenciamento da região ITS, 7 espécies de fungos melanizados que foram identificados já foram citadas na literatura como agentes etiológicos de micoses em humanos: Cladophialophora immunda, Cladosporium sphaerospermum, Cladosporium cladosporioides, Exophiala oligosperma, Exophiala jeanselmei, Hortaea werneckii e Curvularia lunata. A maioria dos isolados foi obtida de espinhos. Alguns destes isolados apresentaram crescimento à temperatura de 37°C. A identificação molecular dos isolados clínicos mostrou similaridade com Fonsecaea pedrosoi, Cladosporium sphaerospermum e Knufia epidermidis. Dessas espécies clínicas somente a espécie C. sphaerospermum foi isolada do ambiente. Este trabalho evidencia que os substratos ambientais estudados são fontes potenciais de infecção de fungos melanizados que podem vir a causar micoses.

Orientador: Marco Aurélio Zezzi Arruda / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Química / Made available in DSpace on 2018-08-20T16:59:51Z (GMT). No. of bitstreams: 1 Gerondi_Fabiana_M.pdf: 2819307 bytes, checksum: 1b5ca46544e4e7b1d9a84e569379ee42 (MD5) Previous issue date: 2012 / Resumo: A técnica espectrometria de absorção atômica utilizando tubo metálico na chama e spray termico (TS-FF-AAS, do inglês, thermospray flame furnace atomic absorption spectrometry) consiste no uso de um tubo metálico sobre a chama do queimador do espectrômetro de absorção atômica, no qual a amostra e introduzida por meio de um capilar cerâmico. No desenvolvimento desse método, as variáveis envolvidas foram estudadas e as condições ótimas consistem no uso de ar a 0,4 mL min como carregador, 200 mL de solução de amostra (em meio ácido) que são injetados no interior de um tubo de Ni sem furos na parte inferior, localizado em uma chama oxidante na proporção de 1,5 L min de acetileno: 12 L min-1 de ar. Nessas condicoes, as menores quantidades que podem ser detectada e quantificada são de 0,15 e 0,50 mg L, respectivamente. Os valores da concentracao de prata em materiais certificados de água (SRM 1643e, Trace Elements in water) e peixe (MA-A-2, Fish Flesh Homogenate) determinados são 1,097 ± 0,062 mg L e 0,094 ± 0,020 mg g , respectivamente, que correspondem a 103 e 94% da quantidade presente no material. Assim, este trabalho propõe uma alternativa para a determinação de prata, na qual não são utilizadas etapas de preconcentração do analito, que são necessárias quando a espectrometria de absorção atômica por chama e utilizada, simplificando o procedimento experimental e diminuindo o tempo de análise e as fontes de erros / Abstract: Thermospray flame furnace atomic absorption spectrometry (TS-FF-AAS) uses a metallic tube on the flame of the atomic absorption spectrometer. The sample is introduced in the tube by a ceramic capillary. In the development of this method, the parameters involved were studied and the optimal conditions consist of using air as carrier at 0.4 mL min, 200 mL of sample (in acid medium) which are injected in a nickel tube without holes in the bottom part and it is located in a oxidant flame with 1.5 L min acetylene: 12 L min air. Under these conditions, the limits of detection and quantification are 0.15 and 0.50 mg L, respectively. The values of silver concentration in certified reference materials of water (SRM 1643e, Trace Elements in Water) and fish (MA-A-2, Fish Flesh Homogenate) are 1.097 ± 0.062 mg L-1 and 0.094 ± 0.020 mg g, respectively, which correspond to 103 and 94% of the certified value. So, this work proposes a simpler and less time consuming alternative to determine silver in environmental samples that do not use preconcentration steps, frequently used when flame atomic absorption spectrometry is used / Mestrado / Quimica Analitica / Mestre em Química