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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
11

Avaliação sazonal dos efeitos do enriquecimento por N e P sobre o perifíton em represa tropical rasa mesotrófica (Lago das Ninféias, São Paulo)

Fermino, Fabiana Schumacher [UNESP] 14 February 2007 (has links) (PDF)
Made available in DSpace on 2014-06-11T19:30:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-02-14Bitstream added on 2014-06-13T19:19:49Z : No. of bitstreams: 1 fermino_fs_dr_rcla.pdf: 1579925 bytes, checksum: 9575963e41a7425b9936cc27d8075a47 (MD5) / O estudo visou (a) identificar o nutriente limitante ao desenvolvimento do perifiton em escala sazonal no Lago das Ninfeias, (b) avaliar os principais atributos da comunidade indicadores desta limitacao, (c) caracterizar as principais alteracoes estruturais da comunidade de algas perifiticas frente ao enriquecimento artificial por nitrogenio e/ou fosforo e hierarquizar os fatores direcionadores dessas modificacoes. Foram realizados enriquecimentos isolados e combinado por N e P em represa rasa mesotrofica em quatro epocas do ano (primavera, verao, outono e inverno). Quatro tratamentos foram delineados utilizando substrato difusor de nutrientes (copos de poliestireno, preenchidos com solucao Agar e nutrientes . controle: sem adicao de nutrientes; N+: 0,75 M; P+: 0,05 M; NP+: adicao combinada dos dois sais, razao molar N:P = 15). A abertura dos copos foi revestida com malha de 20 um como substrato para o perifiton. Coletas foram realizadas aos 15o, 20o, 25o e 30o dias de colonizacao. A maior variabilidade dos dados limnologicos abioticos foi atribuida ao tipo de tratamento, seguida pela escala anual. A ACP indicou forte associacao entre a adicao de fosforo e, principalmente, adicao combinada com as maiores densidades de algas, clorofila-a e conteudo de fosforo do perifiton. O fosforo foi o nutriente limitante ao longo do ano e, dependendo do atributo, foi considerado como nutriente limitante, limitante primario ou co-limitado pelo nitrogenio. Exceto a massa organica, os demais atributos do perifiton foram bons indicadores da limitacao e a composicao quimica do perifiton foi decisiva na definicao desta limitacao. Foram encontrados 203 taxons, com maior representacao da classe Chlorophyceae. A riqueza especifica variou de 33 a 66 e sofreu maior influencia da epoca do ano. / Study aimed at (a) identifying the nutrient limitation to the development of algal periphyton in a seasonal scale at the Ninfeias Reservoir; (b) evaluating the main indicator community attributes of nutrient limitation; and (c) characterizing the main structural modifications underwent by the algal periphytic community caused by the artificial enrichment by nitrogen and/or phosphorus, as well as ranking the driven factors responsible for such modifications. Isolate and combined enrichments were carried out in a shallow, mesotrophic reservoir during four occasions (spring, summer, fall, and winter). Four treatments were designed using diffusing substrate (polystyrene vials filled with agar solution and nutrients . control: no nutrient addition; N+: 0.75 M; P+: 0.05 M; NP+: combined addition of N and P, molar ratio N:P = 15). Vial mouth was covered with a 20 um mesh cloth for periphyton colonization. Samplings were performed on the 15th, 20th, 25th, and 30th colonization days. The greatest abiotic limnological data variation was related to the kind of treatment, followed by the annual scale. PCA indicated a strong association between P and mainly combined P and N addition with the highest algal densities, chlorophyll a, and the P periphyton contents. Phosphorus was the limiting nutrient over the year, and depending on the periphyton attribute, it was either the only limiting nutrient, the primary limiting one, or colimited with N. Except for the organic mass, the remaining periphyton attributes were good nutrient limitation indicators, and periphyton chemical composition was conclusive in defining limitation. Two hundred and three taxa were identified, Chlorophyceae being the dominant algal class. Species richness varied from 33 to 66, being influenced mostly by the time of the year.
12

Avaliação sazonal dos efeitos do enriquecimento por N e P sobre o perifíton em represa tropical rasa mesotrófica (Lago das Ninféias, São Paulo) /

Fermino, Fabiana Schumacher. January 2006 (has links)
Orientador: Denise de Campos Bicudo / Banca: Andréa Tucci / Banca: Liliana Rodrigues / Banca: Raoul Henry / Banca: Orlando Necchi Junior / Resumo: O estudo visou (a) identificar o nutriente limitante ao desenvolvimento do perifiton em escala sazonal no Lago das Ninfeias, (b) avaliar os principais atributos da comunidade indicadores desta limitacao, (c) caracterizar as principais alteracoes estruturais da comunidade de algas perifiticas frente ao enriquecimento artificial por nitrogenio e/ou fosforo e hierarquizar os fatores direcionadores dessas modificacoes. Foram realizados enriquecimentos isolados e combinado por N e P em represa rasa mesotrofica em quatro epocas do ano (primavera, verao, outono e inverno). Quatro tratamentos foram delineados utilizando substrato difusor de nutrientes (copos de poliestireno, preenchidos com solucao Agar e nutrientes . controle: sem adicao de nutrientes; N+: 0,75 M; P+: 0,05 M; NP+: adicao combinada dos dois sais, razao molar N:P = 15). A abertura dos copos foi revestida com malha de 20 um como substrato para o perifiton. Coletas foram realizadas aos 15o, 20o, 25o e 30o dias de colonizacao. A maior variabilidade dos dados limnologicos abioticos foi atribuida ao tipo de tratamento, seguida pela escala anual. A ACP indicou forte associacao entre a adicao de fosforo e, principalmente, adicao combinada com as maiores densidades de algas, clorofila-a e conteudo de fosforo do perifiton. O fosforo foi o nutriente limitante ao longo do ano e, dependendo do atributo, foi considerado como nutriente limitante, limitante primario ou co-limitado pelo nitrogenio. Exceto a massa organica, os demais atributos do perifiton foram bons indicadores da limitacao e a composicao quimica do perifiton foi decisiva na definicao desta limitacao. Foram encontrados 203 taxons, com maior representacao da classe Chlorophyceae. A riqueza especifica variou de 33 a 66 e sofreu maior influencia da epoca do ano / Abstract: Study aimed at (a) identifying the nutrient limitation to the development of algal periphyton in a seasonal scale at the Ninfeias Reservoir; (b) evaluating the main indicator community attributes of nutrient limitation; and (c) characterizing the main structural modifications underwent by the algal periphytic community caused by the artificial enrichment by nitrogen and/or phosphorus, as well as ranking the driven factors responsible for such modifications. Isolate and combined enrichments were carried out in a shallow, mesotrophic reservoir during four occasions (spring, summer, fall, and winter). Four treatments were designed using diffusing substrate (polystyrene vials filled with agar solution and nutrients . control: no nutrient addition; N+: 0.75 M; P+: 0.05 M; NP+: combined addition of N and P, molar ratio N:P = 15). Vial mouth was covered with a 20 um mesh cloth for periphyton colonization. Samplings were performed on the 15th, 20th, 25th, and 30th colonization days. The greatest abiotic limnological data variation was related to the kind of treatment, followed by the annual scale. PCA indicated a strong association between P and mainly combined P and N addition with the highest algal densities, chlorophyll a, and the P periphyton contents. Phosphorus was the limiting nutrient over the year, and depending on the periphyton attribute, it was either the only limiting nutrient, the primary limiting one, or colimited with N. Except for the organic mass, the remaining periphyton attributes were good nutrient limitation indicators, and periphyton chemical composition was conclusive in defining limitation. Two hundred and three taxa were identified, Chlorophyceae being the dominant algal class. Species richness varied from 33 to 66, being influenced mostly by the time of the year / Doutor
13

Regulation and mechanism of mating-type switching in Kluyveromyces lactis

Rajaei, Naghmeh January 2015 (has links)
Transposable elements (TEs) have had immense impact on the structure, function and evolution of eukaryotic genomes. The work in this thesis identified Kat1, a novel domesticated DNA transposase of the hAT family in the yeast Kluyveromyces lactis. Kat1 triggers a genome rearrangement that results in a switch of mating type from MATa to MATα. Furthermore, Kat1 acts on sequences that presumably are ancient remnants of a long-lost transposable element. Therefore, Kat1 provides a remarkable example of the intricate relationship between transposable elements and their hosts. We showed that Kat1 generates two DNA double strand breaks (DSBs) in MATa and that the DDE motif and several other conserved amino acid residues are important for Kat1 cleavage activity. DNA hairpins were formed on one end of the DSBs whereas the DNA between the DSBs was joined into a circle. Kat1 was transcriptionally activated by nutrient limitation through the transcription factor Mts1 and negatively regulated by translational frameshifting. In conclusion, Kat1 is a highly regulated domesticated transposase that induces sexual differentiation.  In another study, we developed an assay to measure switching rates in K. lactis and found that the switching rate was ~6x10-4 events/generation. In a genetic screen for mutations that increased mating-type switching, we found mutations in the RAS1 gene. The small GTPase Ras1 regulates cellular cyclic AMP levels and we demonstrated that Mts1 transcription is regulated by the RAS/cAMP pathway and the transcription factor Msn2. Since Ras activity is regulated by nutrient availability, these data likely explains why nutrient limitation induces mating-type switching.
14

Membrane bioreactor treatment of household light greywater : measurement and effects of phosphorus limitation

Van Epps, Amanda Jane 15 July 2013 (has links)
As water stresses increase across the U.S., interest in household water reuse is growing. Such reuse typically focuses on light greywater, that is all wastewater generated in the house excluding toilet waste and kitchen wastewater. As this practice becomes more widespread, higher level reuse is expected to require greater greywater treatment prior to reuse. Membrane bioreactors (MBRs) are an attractive technology for this application because they offer a robust combination of treatment processes and are already used in some households in countries such as Japan. This research sought to understand the role of phosphorus availability in determining the quality of effluent from MBR treatment of light greywater because phosphorus concentrations are expected to be low with phosphorus phased out of many consumer products. Less than 30 [mu]g/L of dissolved orthophosphate was present in synthetic greywater made from three common household products, and no measurable amount of dissolved orthophosphate was found in real greywater, but low concentrations of particulate phosphate were detected. These concentrations were well below levels believed necessary to achieve full BOD₅ removal in biological treatment. Nevertheless, MBR performance was not adversely affected until no supplemental phosphorus was provided. Measurement of extracellular enzyme activity showed an increase in the ratio of phosphatase activity to total glycosidase activity with declining phosphorus concentration, providing an early indication of nutrient stress before changes in effluent water quality were detected. Removal of three xenobiotic organic compounds (XOCs) in treatment of synthetic greywater was also evaluated under conditions of phosphorous limitation and balance. Abiotic removal mechanisms were not deemed to be important, but removal of methylparaben and sodium lauryl sulfate via biodegradation responded to nutrient limitation similarly to overall COD removal while removal of diethyl phthalate was affected to a greater extent. Measurement of plasmid DNA concentrations was evaluated as a potential indicator of the effect of nutrient limitation on plasmid-mediated biodegradation of XOCs. An overall reduction in the plasmid content was observed in all cases under conditions of phosphorus limitation; however, the extent of reduction was reactor dependent. / text
15

Particulate carbon, nitrogen and phosphorus stoichiometry of south west Florida waters

Murasko, Susan Mary 01 June 2009 (has links)
The southwestern Florida shelf marine environment has often been characterized as oligotrophic, yet these waters can support large, high biomass, persistent phytoplankton blooms, including blooms of the toxin producing dinoflagellate Karenia brevis. Little is known regarding which major nutrient potentially limits primary production in these waters as both inorganic nitrogen and phosphorus concentrations are often near the limits of analytical detection and it is difficult to estimate what percentage of the dissolved organic pool is available for phytoplankton uptake. To assess the nutrient status of phytoplankton populations on the southwest Florida shelf, this project examines the particulate nutrient stoichiometry of ambient phytoplankton assemblages from 1998-2000 as part of the ECOHAB: Florida Program. Particulate C, N, P concentrations and particulate ratios display a large range of values across the West Florida Shelf (WFS). The average particulate stoichiometry is well above the classic Redfield ratio with a geometric mean of 410C:56N:1P. Frequency percentages of particulate ratio values to total sample number binned according to potential nutrient limitation indicate that 39% (C:N) of the data have values suggesting N limitation and that from 88% (N:P) to 95% (C:P) of the data have values which suggest P-limitation. It is difficult to discern whether phytoplankton biomass is truly P-limited as related to the nutrient regime on the WFS or whether detrital contributions, which can potentially be large on this shallow shelf, are skewing the N:P and C:P ratios towards higher values. Errors which could potentially be related to the different methodologies of determining C, N and P concentrations must also be considered when interpreting the particulate nutrient ratios. The data were also analyzed as subsets to determine near-shore to offshore, latitudinal, seasonal, inter-annual and K. brevis bloom versus non-bloom trends. The near-shore to offshore transect indicates decreasing concentrations of particulate C, N, P concentrations and increasing C:N, N:P, C:P ratios with increasing distance offshore. Particulate nutrient concentrations and particulate ratio values are very similar between the Tampa Bay, Sarasota and Fort Meyers transects indicating that these latitudes are not spatially distinct with regards to these variables. There does not appear to be any relationship between the particulate C, N, P concentrations or C:N, N:P, C:P ratios and rainfall as indicated by Spearman Ranking Correlation coefficients. However, there does appear to be monthly trends across the shelf where peak particulate nutrient concentrations and particulate ratio values occur during the spring, summer and fall. The average particulate nutrient concentrations and ratios differ for each year as well as each K. brevis bloom which occurred during the study period. In summary, the particulate C, N, P concentrations and particulate nutrient ratios vary both spatially and temporally on the WFS and are potentially related to the flexibility of phytoplankton uptake kinetics in response to the varying nutrient regimes of the WFS.
16

Benthic Responses to Nitrogen and Phosphorus Deposition on Alpine Ponds in Banff National Park: A Replicated Whole-Ecosystem Experiment

Porter, Lisa, L. Unknown Date
No description available.
17

Light attenuation, phytoplankton and epiphyte diversity as a function of water quality in post flood and recolonising seagrass habitats in Hervey Bay, Queensland.

Deborah Milham Scott Unknown Date (has links)
The quest to discover simple, sensitive and global bioindicators of nutrient enrichment and ecological health continues. The present study is the first to address this quest in Hervey Bay by investigating links between key physico-chemical water quality parameters and two diatom forms. Free floating in the water column as phytoplankton, and attached to seagrass leaves and artificial substrates as microalgal epiphytes, diatoms are assessed for their abundance and diversity along an environmental gradient. Relationships between Secchi depth (Zsd), light attenuation coefficient (Kz) and minimum seagrass compensation depth (Zc) are established for the recovery, growth and survival of aquatic plants following disturbance, with equations derived specifically for this region to allow conversions from Zsd to Kz. These relationships allow predictions to be made and provide a management tool to meet water quality targets and combat elevated nutrient and sediment loads that result from anthropogenic activity and affect all marine life, from microalgae to marine mammals. Environmental links, identified as drivers of change to biodiversity, focus research effort and provide predictive management tools. Consequently, catchment and coastal activities such as dredging, construction and canal development, with the potential to increase nutrient and sediment loads, can be managed and scheduled at times least likely to adversely impact near shore ecosystems and therefore improve the balance between growth and sustainability. Seasonal and zone differences were significant for many of the water quality parameters monitored in Hervey Bay during 1994. Water clarity measures consistently showed significant differences between near and offshore zones (Secchi depth; P<0.01, turbidity; P<0.05 and TSS; P<0.01) as well as oxygen (P<0.01), soluble reactive phosphate (SRP, P<0.05), oxides of nitrogen (NOx, P<0.01), chlorophyll a (Chl a, P<0.01) and pH (P<0.01). Some default trigger values for water quality targets listed by ANZECC/ARMCANZ (2000) for concentrations of key indicators, a threshold for risk of adverse biological effects, were exceeded in dry and flood periods during the 1993 to 1995 survey. The community structure of phytoplankton and seagrass epiphytes were examined by monitoring changes to and 35 seagrass epiphyte species coincident with changes in some key water quality parameters along a water quality gradient. Chl a, Secchi depth (Zsd), soluble reactive phosphate (SRP) and pH were found to be the best subset of water quality parameters to maximise the rank correlation with phytoplankton communities whereas NOx and temperature maximised the rank correlation with the Halophila ovalis seagrass epiphyte communities. These parameters statistically link key water quality parameters to changes in phytoplankton and seagrass epiphyte density and assemblage structure and are supported as significant drivers of change in biodiversity research. Evidence for nitrogen limitation was found in the post flood surveys for phytoplankton and the growth and assemblage structure for seagrass systems and their epiphytes. Seasonal and zone differences in phytoplankton assemblage structure were most apparent with some site influence detected. Cylindrotheca closterium dominated the phytoplankton assemblage structure at the near shore zone while Thalassionema nitzschioides and Chaetoceros sp. differentiated the river from the Waste Water Treatment Plant (WWTP) creek site. Rhizolsolenia and Guinardia sp. were found in significantly higher concentrations at offshore sites (P < 0.01) and Thalassionema nitzschioides (P < 0.01), Thalassiosira sp. (P < 0.01) and Pseudonitzschia sp. (P < 0.05) were found in significantly higher abundance in the near shore zone A Trichodesmium bloom was examined and post flood changes to the phytoplankton assemblage structure associated with increased nutrient loads, reduced water clarity (Secchi depth, TSS and turbidity) and changes to pH and salinity were assessed. The results of analyses of variance provided support to multivariate statistical analyses to identify phytoplankton as a useful and sensitive bioindicator of environmental change. Post flood phytoplankton cell density increased and species diversity rose from 10 to 38 species at the Mary River mouth however, the changes to cell density and assemblage structure were not reflected in Chl a concentrations. The Mary River mouth experienced growth of mainly small phytoplankton species (< 20 µm) while Pulgul Creek, a source of WWTP effluent, experienced a disproportional increase in the larger phytoplankton species (>20µm). The increase in phytoplankton cell density at offshore sites occurred for both large and small species. Phytoplankton species dominance changed at each site during the postflood period. C. closterium remained the dominant species but increased from 34 to 648 cells/mL at Pulgul Creek however, C. similis, the second most dominant species was replaced by Pseudonitszchia sp., whose cell density rose from 2.0 to 320 cells/mL. A recent study also found that Cylindrotheca closterium, Skeletonema costatum, and Cyclotella choctawhatcheeana had strong positive relationships with coastal nutrients and suggested these species be used as potential reliable indicators of eutrophication (Toming and Jaanus, 2007). Some Pseudonitszchia sp. and dinoflagellate species have previously been noted as toxic and Chaetoceros sp. are associated with gill damage and fish kills. Rhizosolenia sp. and the Guinardia species G. flaccida and G. striatula, were indicators for the offshore relatively pristine sites, which may conversely, determine these species as indicators of environmental health in this waterway. Specificity of epiphyte attachment to particular seagrass species were detected and microalgal epiphyte assemblages on Halophila ovalis were identified as useful bioindicators of environmental gradients as this seagrass species is located at intertidal, shallow and deep water sites for a substantial part of the year. H. ovalis also recorded the highest average annual epiphyte loads (3873 ± 1882 cells/mm2) with the diatoms Diatoma vulgare and Cocconeis scutellum representing 19.6% and 17.2% of the total epiphyte cover. Highest cell density was recorded in autumn, followed by summer and winter: spring recorded the lowest epiphyte cover. Micro-algal epiphyte load was assessed as dry weight (g/m2), Chl a (µg/L) and cell density (cells/mm2) on artificial seagrass deployed along a water quality gradient. Equations were derived to describe percentage light transmission as a function of each measure of epiphyte load which allows transformation of data from one unit of expression to another and hence, allow comparison of past, current and future studies. Water column light attenuation was evaluated along a water quality gradient and linked to Secchi depths (Zsd) to derive equations that describe this relationship where for Hervey Bay, when Zsd < 4.26m, Kz = -1.2 ln Zsd + 1.74 and when Zsd ≥ 4.26m, Kz=1.37/Zsd. Species succession, diversity and seagrass growth were monitored for the first time during a recovery phase and limits for the seagrass compensation depth (Zc), were established on a seasonal basis for five seagrass species at four locations within intertidal, shallow and deep water zones in Hervey Bay and the Great Sandy Straits. Conceptual models for each site were constructed to describe habitat characteristics and include nutrient concentrations for SRP, NOx and ammonia (NH4+), Chl a, phytoplankton density, epiphyte density and light requirements measured as light attenuation, total suspended sediment, turbidity and Secchi depth at each location. The study provides a descriptive model for light attenuation and establishes (1) an annual and seasonal baseline water quality data set that characterises the waterways of Hervey Bay and the Great Sandy Straits (2) identification of phytoplankton species in Hervey Bay and their response to post flood changes in water quality (3) phytoplankton density and diversity along an environmental gradient with links established to Chl a, Secchi depth, SRP and pH (4) seagrass epiphyte assemblage structure along an environmental gradient with links established to NOx and temperature (5) conversion factors for epiphyte load expressed as dry weight, Chl a and cell density (6) functions to convert Secchi depth to light attenuation coeffiecients (Kz) (7) seagrass species succession at intertidal, shallow and deep water sites during a recovery growth phase in Hervey Bay (8) seagrass compensation depths (Zc) for five seagrass species at four locations and (9) habitat characterisation for seagrass recovery in Hervey Bay. Identification catalogues for phytoplankton and seagrass epiphytes were prepared with light and electron micrographs to assist future identification studies of diatoms in this region and for other similar biogeographical areas. The management of water quality to reach specific targets requires the capacity to predict seagrass compensation depth (Zc) as a function of water quality. Seasonal and annual light attenuation measurements are derived as a function of water quality at intertidal, shallow and deep water habitats during the colonisation of Zostera Capricornii, Halodule uninervis, Halophila ovalis, Halophila spinulosa and Halophila decipiens.
18

Determination of Nutrient Limitation on Trees Growing in Loxahatchee Impoundment Landscape Assessment (LILA) Tree Islands, Florida

Subedi, Suresh Chandra 28 March 2011 (has links)
The purpose of this study was to determine the general patterns of response by tree species common to Everglades Tree Islands (TI) when conditions limiting optimal growth are improved by fertilization on LILA tree islands. Experiments were conducted on constructed TI in the Loxahatchee Impoundment Landscape Assessment (LILA). Thirty-six trees of two species, Annona glabra and Chrysobalanus icaco, were randomly selected on each of four tree islands. Two tree islands have peat overlying limestone cores and two are composed solely of peat. Each tree was treated with one of three nutrient regimes: +N, +P, or Control (no addition of nutrients). A highly significant P-treatment effect on growth rate, leaf TP and leaf N:P ratio were observed in both species in comparison to Control trees. In contrast, neither of the species responded to N-fertilization. The mass N:P ratios and δ13C in P-treated trees exhibited a positive correlation with Relative Elevation (RE) for both species. These findings suggested that the tree growth at LILA tree islands was P-limited on both substrates (limestone and peat).
19

Begränsande ämne i Bottenhavet : Ett marint system i förändring / Limiting nutrient in the Bothnian Sea : a marine system under change

Johansson, Nadja January 2022 (has links)
During the latest decades, there have been indications that the limiting nutrient are changeable in the Bothnian Sea. The current national regulations for wastewater treatment plants which have coastland towards the Gulf of Bothnia have no regulations regarding nitrogen purification, as these basins traditionally has been seen to have phosphorus-limited conditions. The aim of this study was to investigate how the temporal change of phosphorus and nitrogen has developed during the years 1991-2021 in the Bothnian Sea and if there were any differences between coastal and offshore stations. The ultimate goal was to give an overview of the nutrient situation and develop a guideline whether the current regulations for wastewater treatment plants are sufficient. Three coastal and two offshore stations were studied at the depth 0-10 m by investigating data from national monitoring programs, using trend analysis and comparison of means. The result showed that phosphorus is increasing in all the sampling stations. Consequently, the limited nutrient has evolved towards being more nitrogen-limited. The nitrogen load has instead been larger at the coastal compared to the offshore stations, especially for locations affected by river inlets. However, the conclusion according to this study is that the current regulations are sufficiently. This is due to the more recent shift towards nitrogen-limited conditions at the stations affected by river inlets and the high contribution with phosphorus-rich water from the Baltic Proper, as at the same time wastewater treatment plants is not the greatest contributor to the nitrogen load in the Bothnian Sea.
20

The Role of Multidrug Efflux Pumps in the Stress Response of Pseudomonas aeruginosa to Organic Contamination

Fraga Muller, Jocelyn Lisa 13 September 2006 (has links)
Natural microbial communities are the ultimate drivers of change in any ecosystem. Through chemical contamination of natural environments, these communities are exposed to many different types of chemical stressors; however, research on whole genome responses to this contaminant stress is limited. This research examined the stress response of a common soil bacterium, <i>Pseudomonas aeruginosa</i>, to a common environmental pollutant, pentachlorophenol (PCP). In the first part of the research, it was revealed that nutrient-limited <i>P. aeruginosa</i> is able to respond to PCP with minimal physiological damage due to the upregulation of multidrug efflux pumps. Further study of this PCP-mediated induction of efflux pumps revealed a simultaneous increase in antibiotic resistance. It was discovered that the resistance nodulation-cell division (RND) efflux pump, MexAB-OprM, in particular is responsible for the PCP-induced increase in antibiotic resistance. Both whole cell physiological indicators and whole genome analysis were used to examine the stress response of <i>P. aeruginosa</i> to PCP. Cells were grown in a chemostat at a low growth rate to simulate nutrient-limiting growth in the natural environment. Whole cell acetate uptake rates (WAUR) and viable cell counts as colony forming units (CFU) were determined as cells were exposed to increasing concentration of PCP. At the same time, changes in gene expression were examined by Affymetrix microarray technology. Results showed little change in whole-cell physiology, with no difference in WAUR and only a slight reduction in CFU. However, the microarrays revealed that over 100 genes either increased or decreased expression greater than two-fold due to the PCP exposure. In particular, multiple multidrug efflux genes were upregulated in response to the PCP. The results were validated by real time reverse transcription polymerase chain reaction (RT-PCR) for one of these genes. Further analysis of the effects of MexAB-OprM showed that this particular efflux pump is essential for the response of <i>P. aeruginosa</i> to the toxin PCP. Induction of multidrug efflux pumps is responsible for the development of antibiotic resistance in strains of <i>P. aeruginosa</i>. Therefore, it was investigated whether PCP might induce resistance to a variety of antibiotics. The research was further extended to examine the effect of a variety of organic contaminants on MexAB-OprM efflux and antibiotic resistance development. PCP, 2,4-dinitrophenol, benzoate and Roundup® all induced antibiotic resistance. However, although MexAB-OprM is required for optimal growth in the presence of all chemicals, this particular efflux pump is only involved in increased resistance with PCP. This was confirmed using RT-PCR as <i>mexB</i> expression was induced by PCP, but not by the other three chemicals. A long term generational study on the effects of PCP did not result in a stable antibiotic-resistant phenotype; however, RT-PCR showed that <i>mexB</i> induction is a direct result of PCP exposure and can be reversed by removal of PCP. Together, these results demonstrate the necessity to understand functional responses to contaminant stress. Discovery of direct induction of multidrug efflux pumps and the resulting increase in antibiotic resistance has significant implications for environmental microbiology and public health. This research suggests that organic contamination may result in antibiotic resistance and that antibiotic resistant strains may have a survival advantage in contaminated environments. / Ph. D.

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