10 research outputs found
Isolation and characterization of two cyclin cDNAs from Pisum sativum L.
No full textIn order to investigate the role of cell division in plant development, we isolated two plant genes which encode homologues of animal and yeast cell cycle regulators known as cyclins. Through the use of degenerate primers and the polymerase chain reaction (PCR) we isolated a Pisum sativum sequence which showed homology to the ‘cyclin box°s functional domain found within cyclin proteins. Using this sequence as probe we isolated two different cyclin cDNAs, Pissa;CycA3;1 and Pissa;CycB1;3 from a Pisum sativum L. root tip cDNA library. The deduced amino acid sequences of both cDNAs showed the highest sequence similarity with mitotic cyclins. Analyses of Pissa;CycA3;1 and Pissa;CycB1;3 expression in different tissues, by reverse transcription-polymerase chain reaction (RT-PCR) using primers corresponding to unique regions of their cDNAs, showed their differential expression in relation to cell cycle activity. Furthermore, RT-PCR was used to analyze synchronized root tip cells; results revealed that Pissa;CycA3;1 is preferentially expressed in mid-S (SM) and during late S-G2 (Sl- G2) transition, whereas Pissa;CycB1;3 mRNA is only detectable in Sl and G2 phases
Two and three-color fluorescence flow cytometric analysis of immunoidentified viable bacteria
No full textDNA content, morphometric and molecular marker analyses of Citrus limonimedica, C. limon and C. medica for the determination of their variability and genetic relationships within the genus Citrus
No full textThis work investigated the fingerprinting and phenotyping of Citrus germplasm; species selected were of historical importance belonging to Citrus limonimedica Lush. and its supposed ancestors, along with some other species of the Citrus genus. An integrated approach based on the exploitation of nuclear DNA content, morphological traits and molecular markers, such as RAPD fingerprints and ITS-based SNPs, was employed. We studied a core collection of 54 distinct accessions, including 43 genotypes of the Citrus species (18 species or supposed species) and 11 genotypes of the Poncirus genus, which was used as the reference outgroup. Morphological trait analysis and statistical analysis of DNA content and markers were useful for reconstructing a Citrus phylogeny. In particular, our experiments aimed at estimating the genetic variation within and the genetic relatedness among C limon (L.) Burm., C. limonimedica and C. medica L. to shed light on the hybrid origin hypothesis of C. limonimedica. The results of the multidisciplinary analyses allowed us to confirm a remarkable differentiation between Poncirus and Citrus genera and to highlight a close relationship among the three investigated Citrus species but a distinct difference between these three species and other species in the Citrus genus. RAPD fingerprints and ITS polymorphisms enabled us to point out a variation gradient between lemon and citron, with C. limonimedica as a possible intermediate species. Some accessions of C. medica and C. limonimedica that deviate from such a trend suggest recurrent introgression and/or hybridisation with other species of Citrus
Resolution of viable and membrane-compromised bacteria in freshwater and marine waters based on analytical flow cytometry and nucleic acid double staining.
No full textThe membrane integrity of a cell is a well-accepted criterion for characterizing viable (active or inactive) cells and distinguishing them from damaged and membrane-compromised cells. This information is of major importance in studies of the function of microbial assemblages in natural environments, in order to assign bulk activities measured by various methods to the very active cells that are effectively responsible for the observations. To achieve this task for bacteria in freshwater and marine waters, we propose a nucleic acid double-staining assay based on analytical flow cytometry, which allows us to distinguish viable from damaged and membrane-compromised bacteria and to sort out noise and detritus. This method is derived from the work of S. Barbesti et al. (Cytometry 40:214-218, 2000) which was conducted on cultured bacteria. The principle of this approach is to use simultaneously a permeant (SYBR Green; Molecular Probes) and an impermeant (propidium iodide) probe and to take advantage of the energy transfer which occurs between them when both probes are staining nucleic acids. A full quenching of the permeant probe fluorescence by the impermeant probe will point to cells with a compromised membrane, a partial quenching will indicate cells with a slightly damaged membrane, and a lack of quenching will characterize intact membrane cells identified as viable. In the present study, this approach has been adapted to bacteria in freshwater and marine waters of the Mediterranean region. It is fast and easy to use and shows that a large fraction of bacteria with low DNA content can be composed of viable cells. Admittedly, limitations stem from the unknown behavior of unidentified species present in natural environments which may depart from the established permeability properties with respect to the fluorescing dyes
Rapid flow cytometric immunodetection of bacteria to monitor aquatic environments
No full textInternational audienceno abstrac
Rapid flow cytometric immunodetection of bacteria to monitor aquatic environments
No full textInternational audienceno abstrac
Estudo da minimização da produção de lodo em sistemas de lodos ativados para esgotos sanitários
No full textTese (doutorado) - Universidade Federal de Santa Catarina, Centro Tecnológico, Programa de Pós-Graduação em Engenharia Ambiental, Florianópolis, 2015.A produção de lodo em sistemas de lodos ativados é um grande desafio na gestão de estações de tratamento de esgotos (ETE), tanto pela crescente população servida por sistemas de coleta e tratamento de efluentes, quanto pelas regulamentações ambientais mais restritivas com relação à sua disposição final. Nesse sentido, a exploração e o desenvolvimento de processos que promovam a redução da produção deste resíduo têm sido muito incentivados. Citam-se, por exemplo, os processos baseados em teorias como a lise celular/crescimento críptico, o metabolismo desacoplado e o decaimento endógeno. O presente estudo teve como objetivo identificar os processos mais promissores para redução da produção de lodo em sistemas de lodos ativados, baseado na sua eficiência e interferência no tratamento do efluente (em escala bancada), estudando seus mecanismos de ação e verificando as potenciais condições de otimização do processo (em escala real). Os resultados obtidos em escala de bancada na UFSC, referentes à aplicação de métodos físico-químicos e biológicos, revelaram que algumas técnicas já consolidadas mundialmente como a cloração e a desintegração ultrassônica apresentam eficiência satisfatória, com redução de 25 a 55% da produção de lodo. No entanto, provocam alterações na qualidade do lodo, como a proliferação de organismos filamentosos, diminuindo o desempenho do tratamento. O processo oxic-settling-anaerobic (OSA) (transição entre condições aeróbias/anaeróbias do lodo) foi a alternativa mais eficiente, com redução do lodo excedente entre 50 a 60%, apresentando inclusive melhoras na qualidade do efluente, com eficiências superiores a 90% para DBO e um aumento de 50% na remoção de amônia quando comparado ao sistema controle. O monitoramento de uma estação de tratamento de esgotos em escala real, utilizando um reator anaeróbio (sistema tipo OSA), em paralelo ao processo de lodos ativados, para a redução de lodo excedente, permitiu verificar o rendimento do processo, bem como possibilitou identificar parâmetros chaves para se alcançar a redução de lodo, dentre os quais estão a taxa de recirculação do lodo (recomendada como sendo de 10%) e a idade do lodo global do sistema. Os resultados indicaram que a redução da produção de lodo varia entre 10 a 30%, e pode estar associada à desaceleração da cinética de crescimento da biomassa, juntamente com a solubilização dos compostos orgânicos em condições anaeróbias, enquanto o decaimento e a lise celular ocorrem principalmente em condições aeróbias. A alternância entre condições aeróbias/anaeróbias pode ser considerada o ponto chave para a redução de lodo nos sistemas do tipo OSA.Abstract : The amount of excess sludge production in activated sludge system is one of the most critical challenges in the field of sewage treatment. Considering this amount is set to rise, once the preservation of water sources and the more stringent effluent regulations have increased the population connected to the sewage treatment network. Thus, various approaches to reduce the production of excess sludge in the wastewater treatment plants (WWTP), such as lyses/cryptic growth, uncoupled metabolism and endogenous decay, have been encouraged. This study aimed (1) identify the more promising sludge reduction process based on its capacity to maintain the effluent quality and the sludge properties (bench scale experiment); (2) study the reduction mechanisms and verify the potential operational conditions that could optimize the sludge reduction process (real scale experiment). The bench scale experiment showed that the physical-chemical sludge reduction alternatives, such as chlorination and ultrasonic disintegration, whilst presented good sludge reduction (reduction of 25 to 55% on sludge production), these processes triggered changes on sludge quality resulting in a decline on treatment performance. OSA (oxic-settling-anaerobic) process was the most efficient sludge reduction alternative, with 50 ? 60% of sludge reduction, and the process also presented an improvement on effluent quality, BOD removal efficiencies remained over 90% and increase 50% of the nutrient removal efficiencies compared to control system. The real scale experiment monitored a modified activated sludge system based on OSA-like process. The process presented a sludge reduction of 10 ? 30%, which could be attributed to a decrease on biomass growth kinetics in association with organic matter solubilisation under anaerobic conditions. The sludge decay and cellular lyses occurred mainly under aerobic conditions. The transition from anaerobiosis to aerobiosis appeared one of the key points of sludge reduction in the OSA-like process and the repeated alternation between aerobic and anaerobic conditions could reduce the overall biomass even if the cryptic-growth is included
Enhancing the removal of a diverse range of hazardous chemicals from wastewaters
No full textDue to increasingly stringent legislation covering the discharge of hazardous chemicals into
the environment, existing wastewater treatment processes need to be upgraded for their
removal. This thesis explores the removal of a diverse range of hazardous chemicals during
secondary wastewater treatment with the overall aim of enhancing their removal
simultaneously by activated sludge.
Previous research in this field has made the broad comparison of full-scale activated sludge
plants (ASPs) which receive varying influent sewage compositions and flow. Consequently,
assessing the direct impact of process operation to hazardous chemical removal has been
difficult. In this study, the independent impact of the process variables solids retention time
(SRT) and hydraulic retention time (HRT) were examined using a pilot-scale ASP. To
measure ASP resilience for the removal of a wide range of hazardous chemicals of varied
chemistry and preferred removal pathways steroid estrogens, nonylphenolic surfactants and
metals were monitored. Cont/d
