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Detection of cyanotoxins (microcystins/nodularins) in hepatic tissues and epidermal mats of stranded bottlenose dolphins (Tursiops truncatus) in Northeast FloridaBrown, Amber 01 January 2018 (has links)
The St. Johns River (SJR; Jacksonville, FL, USA) is a large, brackish, estuarine system characterized by considerable anthropogenic pollution, recurrent harmful algal blooms (HABs), and diverse toxin-producing cyanobacteria. The most prevalent toxins in SJR water samples are microcystins/nodularins (MCs/NODs). Additionally, the SJR provides critical habitat for a genetically and behaviorally distinct estuarine community of bottlenose dolphins (Tursiops truncatus) that routinely uses and strands in low mesohaline and oligohaline areas of the river. This population has been subject to two unusual mortality events (UME) since 2010 and has since been described as having substantial declines in population health, characterized by widespread dermatitis and emaciation. Additionally, three dolphins have been recovered from low salinity habitats with epidermal algal mats. Because dolphin illness and strandings overlapped temporally and spatially with confirmed cyanobacterial blooms in the SJR, there is concern that estuarine dolphin health may be declining due to exposure to toxic cyanobacteria and HAB events. Specific to this study, the SJR estuarine community was considered a high-risk group for cyanotoxin exposure in relation to coastal animals. This study analyzed all available hepatic tissues for estuarine dolphins, and used samples from coastal individuals that stranded outside of the known cyanotoxin bloom season as controls. Three analytical methods were used to determine MCs/NODs presence in dolphin liver and epidermal algal mat samples. An Adda ELISA and LC-MS/MS were used to determine free MCs/NODs presence while the MMPB technique was used to determine total (bound and free) concentrations and as confirmatory analyses. ELISA analyses produced high values that were not supported by concurrent LC-MS/MS or MMPB analyses, indicative of false positives. MMPB testing resulted in low-level total MCs/NODs detection in some specimens. Results indicate that both estuarine and coastal dolphins are exposed to MCs/NODs, with potential toxic and immune health impacts.
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Die raum-zeitliche Variation von Microcystis spp. (Cyanophyceae) und Microcystinen in der Talsperre Quitzdorf (Sachsen)Ihle, Tilo 26 June 2008 (has links) (PDF)
Cyanobakterien bilden zahlreiche bioaktive Substanzen mit zum Teil humantoxischer Relevanz. Nicht selten spielen dabei zyklische Peptide, zu denen unter anderem die Microcystine (MCYST) gehören, eine Schlüsselrolle. MCYST werden u.a. von Microcystis KÜTZING EX LEMMERMANN 1907 gebildet. Erkenntnisse zur ökophysiologischen Funktion der MCYST, die zweifelsfrei bei den Produzenten selbst zu suchen ist, liegen bisher kaum vor. Mit Hilfe von Freilanduntersuchungen sollten im Rahmen der vorliegenden Arbeit Kenntnisse zu einer möglichen ökologischen Funktion der MCYST erweitert und vertieft werden. Grundlage stellte dabei die Phänologie von Microcystis als einer der bedeutendsten limnischen MCYST-Produzenten dar. Microcystis zeigt im Freiland einen charakteristischen annuellen Lebenszyklus mit benthisch-pelagischer Kopplung. Ziel der vorliegenden Arbeit war es, die phänologischen Phasen des Lebenszyklus von Microcystis im Freiland zu differenzieren sowie die Dynamik der MCYST während dieser Phasen kompartimentübergreifend gesamtheitlich zu erfassen. Über eine MCYST-Massenbilanzierung sollen anschließend die dem annuellen Zyklus zugrundeliegenden Teilprozesse quantifiziert und zusammengeführt werden. Vordergründiges Anliegen war es, Phasen einzugrenzen, bei denen MCYST möglicherweise eine ökophysiologische Funktion haben könnte. Der annuelle Lebenszyklus von Microcystis wurde anhand von Biomasseänderungen am Sediment und im Pelagial der TS Quitzdorf in die phänologischen Phasen Überwinterung, Reinvasion, pelagisches Wachstum und Sedimentation unterteilt: Intakte, im Herbst aus dem Freiwasser aussedimentierte, Microcystis-Kolonien überwintern am Sediment und steigen im Frühjahr und Frühsommer zurück ins Freiwasser auf. Dort erfolgt der Wachstumsprozess, dem sich im darauffolgenden Herbst erneut ein Zusammenbruch und die Sedimentation der Freiwassergemeinschaft anschließt. Die benthisch-pelagische Kopplung wirkt dabei als interannuelles Bindeglied. Zwischen dem annuellen Lebenszyklus von Microcystis und der MCYST-Dynamik wurde eine enge Bindung nachgewiesen: Änderungen der absoluten MCYST-Konzentrationen während der Übergangsphasen Aufstieg (Frühjahr) und Sedimentation (Herbst) zeigen, dass MCYST mit den aufsteigenden bzw. aussedimentierenden Microcystis-Kolonien aus dem bzw. in das Sediment ‚transportiert’ werden. Ausschließlich während der pelagischen Phase, die sich dem Reinvasionsprozess anschließt, kommt es in Abhängigkeit vom Wachstum der Produzenten und deren Sukzession zur Neubildung von MCYST. Während den Wintermonaten wurden MCYST am Sediment intrazellulär ‚konserviert’. Der Verlauf der pelagischen MCYST-Konzentration wurde mit Hilfe eines Wachstumsmodells nachgebildet. In dieses Modell wurde die genetische Variabilität der MCYST-Produzenten sowie eine mögliche physiologische Steuerung der MCYST-Synthese über die Verfügbarkeit des anorganischen Kohlenstoffs integriert. Der prinzipielle Verlauf zeigte dabei weitestgehend Koinzidenz zwischen den real gemessenen und den simulierten MCYST-Konzentrationswerten. Abweichungen zwischen beiden konnten mit Hilfe des gesamtheitlich kompartimentübergreifenden MCYST-Bilanzierungsansatzes – in erster Linie über benthisch-pelagische Kopplungsprozesse – plausibel erklärt werden. Der Habitatwechsel ist für Microcystis prinzipiell mit Verlusten (Seneszenz/Lyse oder möglicherweise Apoptose) verbunden, sowohl für MCYST-Produzenten und Nichtproduzenten. Die auffallende Stabilität der benthischen MCYST-Zellquote während der Überwinterung gibt Grund zur Annahme, dass eine Funktion von MCYST am/im Sediment eher unwahrscheinlich ist. Da MCYST über derart lange Zeiträume am Sediment intrazellulär ‚konserviert’ werden, ist eine Bedeutung der MCYST während der Reinvasionsphase und in der frühen pelagischen Phase nicht auszuschließen. Im Speziellen wurde eine mögliche ökologische Funktion von MCYST in Zusammenhang mit der Variation der Koloniegröße bzw. dem epiphytischen Bewuchs von Microcystis-Kolonien mit Pseudanabaena mucicola geprüft: Aus dem Zusammenhang zwischen extra-/intrazellulärer MCYST-Konzentration und der Microcystis-Koloniegrößenverteilung waren keine konsistenten Schlussfolgerungen abzuleiten, welche auf eine Steuerung der Koloniebildung durch MCYST deuten. Vor dem Hintergrund, dass MCYST keinen nachweislich allelopathischen Effekt auf den Epibionten Pseudanabaena mucicola ausüben, wurde postuliert, dass zwischen dem beobachteten epiphytischen Besiedlungs-/Verteilungsmuster und der MCYST-Produktion ein indirekter Zusammenhang besteht, welcher die zeitweise Einnischung von Pseudanabaena mucicola auf Microcystis-Kolonien ermöglicht. Die Ergebnisse der vorliegenden Untersuchung lassen weder unmittelbar noch mittelbar eine Variabilität der ökophysiologischen Bedeutung von MCYST, die im Zusammenhang mit der raum-zeitlichen Verteilung potentieller Produzenten steht, erkennen. Eine divergierende Funktion der MCYST auf intra- bzw. extrazellulärer Ebene kann nicht zwingend ausgeschlossen werden. Die Mehrzahl der aus der MCYST-Phänologie und MCYST-Bilanzierung abzuleitenden Schlussfolgerungen deutet allerdings eher auf eine Funktion auf (intra-)zellulärer Ebene hin, wie etwa die Effizienzsteigerung des Kohlenstoffmetabolismus (d.h. der intrazellulä-ren Akkumulation anorganischen Kohlenstoffs) während der pelagischen (Wachstums-)Phase der Produzenten.
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Rôle de l’azote dans la structure et la fonction des communautés de cyanobactéries toxiquesMonchamp, Marie-Eve 03 1900 (has links)
Dans cette étude de trois lacs sujets aux efflorescences de cyanobactéries, nous
avons examiné la diversité des bactéries diazotrophes et des cyanobactéries toxiques. Nous avons tenté de définir les facteurs environnementaux influençant la composition des communautés phytoplanctoniques, la concentration ainsi que la composition des
microcystines (MCs). Nous avons émis l’hypothèse que l’azote jouerait un rôle majeur dans le façonnement des communautés cyanobactériennes et influencerait la concentration et composition des MCs. Des concentrations de cette toxine ainsi que le gène mcyE codant pour l’enzyme microcystine synthétase ont été détectés à chaque échantillonnage dans tous les lacs. L’azote, particulièrement sous sa forme organique dissoute (AOD) ainsi que la température de l’eau étaient les facteurs environnementaux expliquant le mieux les concentrations des MCs, tandis que la biomasse de Microcystis spp. était globalement le meilleur prédicteur. Le gène nifH codant pour l’enzyme nitrogénase (fixation d’azote) a
aussi été détecté dans chaque échantillon. Malgré les concentrations faibles en azote
inorganique dissous (AID) et les densités importantes d’hétérocystes, aucun transcrits du gène n’a été détecté par réverse-transcription (RT-PCR), indiquant que la fixation d’azote n’avait pas lieu à des niveaux détectables au moment de l’échantillonnage. De plus, le pyroséquençage révèle que les séquences des gènes nifH et mcyE correspondaient à
différents taxons, donc que les cyanobactéries n’avaient pas la capacité d’effectuer les deux fonctions simultanément. / In this study of three eutrophic lakes prone to cyanobacterial blooms, we examined
the diversity of diazotrophic bacteria and toxic cyanobacteria. We evaluated the
environmental factors effects on the community composition, the concentration and
composition of the family of toxins microcystins (MCs). Since the assimilation of nitrogen and the synthesis of MCs in cyanobacteria are thought to be under the same control of the NtcA transcriptor, we hypothesised that nitrogen played a major role in shaping cyanobacterial communities and influenced indirectly the concentration and composition of MCs. The mcyE gene coding for the microcystin synthetase enzyme and MCs
concentrations were detected at each sampling date in all lakes. Nitrogen, particularly under its organic dissolved form (DON) as well as water temperature were the environmental factors explaining the most variation in MC concentration although Microcystis spp. biomass was overall the best predictor. The nifH gene coding for the nitrogenase enzyme (N2-fixation) was also detected at all times. Even though the concentrations of dissolved inorganic nitrogen were relatively low, and that heterocysts were present in high densities, no nifH transcripts were detected by RT-PCR, indicating that no N2-fixation was going on
at detectable levels at the time of sampling. Moreover, pyrosequencing revealed that sequences of the genes nifH and mcyE corresponded to different taxa, thus cyanobacteria did not have the capacity to perform both functions simultaneously.
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Comunidades de cianobacterias bentónicas, producción y liberación de microcistinas en el río Muga (NE Península Ibérica)Vassal'lo Saco, Jara 24 February 2010 (has links)
El estudio de las comunidades de cianobacterias en la cuenca del río Muga permite profundizar en la búsqueda de factores que favorezcan o inhiban la acumulación intracelular de microcistinas y su liberación al medio externo en condiciones oligotróficas o con leves signos de eutrofia, al tiempo que permiten aportar nuevos conocimientos que ayuden a mejorar el entendimiento de los mecanismos y procesos que participan en la dinámica de las comunidades de cianobacterias bentónicas y las microcistinas asociadas en ecosistemas fluviales mediterráneos. Se ha realizado un estudio de la estructura y variabilidad de las comunidades de cianobacterias durante un ciclo anual completo mediante muestreos mensuales y muestreos con menor periodicidad durante un segundo ciclo posterior. Dichos muestreos han permitido observar diferentes patrones espacio-temporales a lo largo de todo el período de estudio en los 4 puntos de muestreo seleccionados en la cuenca del río Muga. Estos 4 tramos incluyen 1 punto regulado:aguas abajo del embalse de Boadella, y 3 sin regular:2 cabeceras y un punto aguas abajo de la población de Albanyà. Se han identificado un total de 34 especies de cianobacterias bentónicas en las 4 zonas estudiadas, predominando siempre especies de aguas limpias o poco alteradas. Dos especies, Rivularia biasolettiana (zonas más limpias y expuestas) y Phormidium retzii (sonas esciáfilas o con alta velocidad de corriente), resultan dominantes en gran parte de ellas. Se han reconocido un total de 6 microcistinas acumuladas intracelularmente en la comunidad bentónica de cianobacterias, identificando 3 de ellas: MC-RR, MC-LR y MC-YR. La concentración de microcistinas intracelulares ha disminuído a lo largo del eje principal del río Muga, determinada por diferentes factores físicos, químicos y biológicos. A nivel general, la acumulación total de microcistinas intracelulares ha estado asociada a un aumento de la temperatura del agua, un descenso en la diversidad de la comunidad de cianobacterias y pequeños incrementos de la concentración de fósforo disuelto en el agua. En cuanto a la liberación de microcistinas, ha estado vinculada a distintas perturbaciones soportadas por la comunidad cianobacteriana, tanto asociadas a factores físicos naturales como antrópicos. / The study of cyanobacteria communities in the Muga River basin enables further search for factors that promote or inhibit the intracellular accumulation of microcystins and their external release in oligotrophic or sligthly eutrophic conditions. At the same time, it provides new knowlodge to improve the understanding of the mechanisms and proccesses involved in the community dynamics of benthic cyanobacteria and associated microcystins in Mediterranean River ecosystems. A two-year study of the structure and variability of cyanobacterial communities was carried out: monthly sampling during the first annual cycle and lower frequency sampling during the second cycle. This allows observing the different spatial-temporal patterns over the study period in the 4 selected sampling points in the Muga River basin. These 4 points comprise 1 regulated point downstream of the Boadella reservoir, and 3 unregulated points, namely 2 headers and a thrid one downstream of Albanyà village. A total of 34 species of benthic cyanobacteria were identified in the 4 studied areas. The predominant species were linked to clear waters or slightly altered areas. 2 species were dominant in most of them: Rivularia biasolettiana (in cleaner and exposed areas) and Phormidium retzii (in unexposed areas or with high flow velocity).In the benthic community of cyanobacteria, a total of 6 intracellular microcystins were detected. 3 of them were identified: MC-RR, MC-LR and MC-YR. The accumulation of intracellular microcystins decrease along the stream as determined by different physical, chemical and biological factors. At a general level, the total accumulation of intracellular microcystins was associated with an increase in water temperature, a decrease of benthic cyanobacterial diversity and small increases in the concentration of dissolved phosphorus in the water. On the other hand, microcystins release was linked to the effects of different physical disturbances on the cyanobacterial community, wether of natural or anthropical origin.
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The destruction of cyanobacterial toxins with oxidants used in drinking water treatmentBrooke, Samuel January 2009 (has links)
Saxitoxins were extracted from a bloom of toxic Anabaena circinalis and used to spike treated water from Hope Valley Reservoir (HVTW) and Milli-Q water. The waters were treated with ozone using the batch method and saxitoxin levels were measured in the samples using HPLC. The results for oxidation of saxitoxins in Milli-Q water versus HVTW show that despite the presence of natural organic matter(NOM) and the production of vastly different ozone residuals, there was a similar removal of all saxitoxins in both waters. The results show that high concentrations of saxitoxins were present in solution after ozonation with doses and contact times typically used in water treatment. Relating the toxin destruction to ozone residual showed that even with a residual ozone concentration of 0.8 mg/L after 10 minutes contact in HVTW, over 60% of the initial saxitoxin content was still present in the samples. The presence of an ozone residual in the water could not be related to saxitoxin destruction and it appeared that saxitoxin removal occurred more rapidly when ozone was consumed rather than stabilised in solution. The results indicate that the mechanism for toxin removal is probably based on the reaction with a hydroxyl radical species as the oxidant rather than molecular ozone. The results obtained during these experiments indicate that ozone is not an effective oxidant for this class of compound. A range of ozone doses were applied to two different treated reservoir waters that had been spiked with microcystins LA (mLA) and LR (mLR). At the ozone dose where a residual was first measured in the sample after 5 minutes exposure time, no microcystins were detected by HPLC in either water. The removal of mLA and mLR was identical in all samples. The absence of mLA and mLR by HPLC was supported by a loss of toxicity using a highly sensitive and specific bioassay (PP2A) and by in vivo studies in mice. In both waters microcystins were removed with an ozone dose typical of that used in drinking water treatment. The results indicate that conventional ozone treatment was effective in removing hepatotoxicity at microcystin levels greater than those likely to be found in drinking water. Two waters were sampled from reservoirs in South Australia. One was collected directly from Happy Valley Reservoir (HVRW) and the other from Myponga Reservoir after treatment but before chlorination (MFCW). They were spiked with mLA and mLR and chlorinated to measure toxin removal and chlorine consumption using the CT concept. In MFCW at pH 7 there was a better removal of both mLA and mLR than in HVRW at pH 8.1. There was also a lesser effect from water temperature upon toxin removal in MFCW. Microcystin LA was less easily removed than mLR at both temperatures in both waters. For HVRW, at the higher pH, this required an initial dose of 7 mg/L of chlorine which corresponded to a CT of around 70 min.mg/L. If the water temperature was reduced to 6??C then under these conditions there would still be 40% of the initial concentration of mLA and mLR present in this water. At this temperature a final chlorine residual of 3.5 mg/L after 30 minutes, requiring a chlorine dose of 8mg/L and corresponding to a CT of about 95 min.mg/L, was required to reduce microcystin levels below the WHO guidelines. This implies that in colder climates the application of chlorine for microcystin removal may require elevated chlorine doses and CT values. Arrhenius activation energies were calculated for mLA and mLR in both waters, revealing different Ea values for both toxins. Due to the complexity of the reactions and the possible effects of pH in solution, this system was considered too complicated to be described by the Arrhenius equation. NOM was collected from Myponga Reservoir in South Australia using magnetic ion exchange (MIEX??) resin. The collected NOM was desorbed and separated into fractions of different molecular weight and character using ultrafiltration and mixed resin ion exchange. At approximately 5 mg/L dissolved organic carbon (DOC) the measured apparent second order rate constant (kapp) for mLA and mLR removal was fairly similar in both the high molecular weight fraction (designated F3), and the intermediate high molecular weight fraction (designated F2). The low molecular weight fraction (designated EN) had slightly higher kapp values as would be expected due to the less reactive nature of the NOM in this fraction. This meant more chlorine was available to react with microcystins in this fraction. Fractions F3 and F2 produced similar kapp values to those from the parent water source following treatment, indicating the similar reactivity of these NOM fractions at comparable DOC levels. Increasing the DOC concentration in the F2 fraction increased kapp for both mLA and mLR due to the additional chlorine needed to react with the additional NOM present. The results showed that pH, temperature and DOC concentration have a higher impact upon chlorination rates, and the efficiency of toxin removal, than NOM character alone. In general it is assumed that chlorine will be more effective at removing toxins in water with a low SUVA and low specific colour as these indicate less 'reactive' NOM in the water. The results of this study show that toxin removal was more effective in the EN fraction as indicated by the higher kapp. This fraction also had the lowest SUVA and lowest specific colour which supports the generally held view in water treatment. Relating the toxin removal to chlorine residual in these reconstituted fractionated NOM samples, indicated that a residual of around 1.5 mg/L after 30 minutes contact was generally adequate to remove all toxins in water with a DOC level of around 5 mg/L. This is consistent with the results obtained in real waters, where at 20??C a chlorine residual of 2 mg/L was found to be sufficient for removal of both mLA and mLR.
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Avaliação da toxicidade de Microcystis aeruginosa e de florações naturais de cianobactérias de reservatórios do rio Tietê, SP / Toxicity evaluation of Microcystis aeruginosa cultures and natural cyanobacteria blooms from reservoirs of Tietê river, SPRenata Akemi Takenaka 16 March 2007 (has links)
Os efeitos de cianobactérias sobre organismos aquáticos planctônicos foram avaliados, visando caracterizar e quantificar as cianotoxinas e determinar a toxicidade de uma linhagem em cultura monoespecífica e de florações naturais de reservatórios do rio Tietê, SP. Assim, cultivou-se uma linhagem (NPLJ-4) de Microcystis aeruginosa, reconhecidamente tóxica, em meio ASM-1 a 25°C e fotoperíodo de 12h luz/12h escuro em câmara incubadora, avaliando-se sua toxicidade em diferentes estágios do crescimento populacional (meio e final da fase exponencial, fase estacionária e fase senescente), por meio de testes ecotoxicológicos com os organismos-teste Ceriodaphnia dubia e C. silvestrii. Esses testes foram realizados de acordo com normas padronizadas pela ABNT, sendo utilizados também para avaliar a toxicidade das florações naturais e a eficiência de diferentes processos de tratamento de água na remoção de células, microcistinas e subprodutos de cianobactérias. Os resultados indicaram aumento na concentração de microcistinas com o crescimento populacional da cianobactéria. Os extratos na fase estacionária tiveram menor toxicidade, enquanto nas demais fases causaram efeito tóxico agudo, resultando em valores de CE50; 48h de: 1,4 - 4,7 × \'10 POT.6\' cel/mL (meio fase exponencial), 1,6 - 8,7 × \'10 POT.6\' cel/mL (final fase exponencial), 7,5 - 14,1 × \'10 POT.6\' cel/mL (fase estacionária) e 1,9 - 4,6 × \'10 POT.6\' cel/mL (fase senescente) para C. dubia; e 1,9 - 5,4 × \'10 POT.6\' cel/mL (meio fase exponencial), 1,6 - 10,9 × \'10 POT.6\' cel/mL (final fase exponencial), 10,2 - 15,4 × \'10 POT.6\' cel/mL (fase estacionária) e 2,0 - 4,2 × \'10 POT.6\' cel/mL (fase senescente) para C. silvestrii. Células livres de Microcystis (M. aeruginosa, M. panniformis e M. protocystis) e Pseudanabaena mucicola foram as cianobactérias dominantes nas florações dos reservatórios de Barra Bonita e células livres de Microcystis (M. aeruginosa e M. panniformis), no de Promissão. A dominância das cianobactérias em ambos os reservatórios pode estar relacionada a períodos de estabilidade da coluna de água, razões N/P de 8 - 13 (Barra Bonita) e 19 - 20 (Promissão) na superfície, temperatura da água de 19 - 30°C (Barra Bonita) e 26 - 28°C (Promissão) e disponibilidade de nutrientes (0,05 - 0,26 mg/L de fósforo total para Barra Bonita e 0,01 - 0,05 mg/L P-total para Promissão), devido ao grau de trofia dos reservatórios. A água de ambos os reservatórios, coletada durante as florações, apresentou toxicidade aos dafinídeos, sendo que a água de Barra Bonita foi mais tóxica do que a de Promissão. Todos os extratos brutos de material oriundo das florações naturais apresentaram microcistinas (239 - 1647 µg/L para Barra Bonita e 192 - 1295 µg/L para Promissão) e causaram toxicidade aguda aos dafinídeos, com valores de CE50; 48h de: 87 - 282 mg/L (Barra Bonita) e 146 - 428 mg/L (Promissão) para C. dubia, e 98 - 546 mg/L (Barra Bonita) e 110 - 391 mg/L (Promissão) para C. silvestrii. Concentrações dos extratos a partir de 80 mg/L (Barra Bonita) e 100 mg/L (Promissão) afetaram adversamente a sobrevivência e a reprodução dos dafinídeos. Os resultados mostram riscos à biota natural e à saúde humana, bem como comprometimento dos usos múltiplos dos reservatórios, exigindo ações remediadoras e, sobretudo, preventivas para conter o processo de eutrofização. / The effects of cyanobacteria upon aquatic organisms were evaluated, aiming to characterize and quantify the toxins of both, a monospecific cyanobacterial culture and material from natural blooms occurring in the reservoirs of Tietê river, SP. The already known toxic strain NPLJ-4 of Microcystis aeruginosa was cultured in ASM-1 medium at 25 Celsius degrees and 12h light/12h dark in the incubator, in order to evaluate its toxicity at different stages of the culture growth by ecotoxicological tests using the cladocerans Ceriodaphnia dubia and C. silvestrii as test-organisms. These tests were carried out according to the procedures standardized by ABNT, in order to evaluate also the toxicity of natural blooms and the efficiency of different water treatment processes in removing cells, microcystins and by-products of cyanobacteria. The results obtained indicated an increase in the concentration of microcystins along the cyanobacterial culture growth. Extracts from the stationary phase of the culture were less toxic compared with those from the other phases which had acute toxicity and adversely affected cladoceran survival, resulting in EC50; 48h values of 1,4 - 4,7 × \'10 POT.6\' cells/mL (middle exponential phase), 1,6 - 8,7 × \'10 POT.6\' cells/mL (final exponential phase), 7,5 - 14,1 × \'10 POT.6\' cells/mL (stationary phase) and 1,9 - 4,6 × \'10 POT.6\' cells/mL (senescent phase) for C. dubia; and 1,9 - 5,4 × \'10 POT.6\' cells/mL (middle exponential phase), 1,6 - 10,9 × \'10 POT.6\' cells/mL (final exponential phase), 10,2 - 15,4 × \'10 POT.6\' cells/mL (stationary phase) and 2,0 - 4,2 × \'10 POT.6\' cells/mL (senescent phase) for C. silvestrii. Cells of Microcystis (Microcystis aeruginosa, M. panniformis and M. protocystis) and Pseudanabaena mucicola were cyanobacteria species dominant in the Barra Bonita reservoir and cells of Microcystis (M. aeruginosa and M. panniformis), in the Promissão reservoir. The dominance of cyanobacteria in both studied reservoirs was related to the stability of the water column, N/P ratios of 8 to 13 (Barra Bonita) and 19 to 20 (Promissão), high water temperatures (19 - 30°C for Barra Bonita and 26 - 28°C for Promissão) and high nutrient availability (0,05 - 0,26 mg/L total phosphorus for Barra Bonita, and 0,01 - 0,05 mg/L total P for Promissão) as a consequence of the trophic state of the reservoirs. The water from Barra Bonita reservoir during the cyanobacterial blooms was more toxic to daphnids than that from Promissão reservoir. Crude extracts from all cyanobacteria blooms tested presented microcystins (239 - 1647 µg/L for Barra Bonita and 192 - 1295 µg/L for Promissão) and caused acute toxicity to daphnids, resulting in EC50; 48h values of 87 - 282 mg/L (Barra Bonita) and 146 - 428 mg/L (Promissão) for C. dubia, and 98 - 546 mg/L (Barra Bonita) and 110 - 391 mg/L (Promissão) for C. silvestrii. Crude extracts concentrations above 80 mg/L to Barra Bonita and 100 mg/L to Promissão adversely affected the survival and reproduction of daphnids. The results obtained evidenced the risks to the natural biota and possibly to the human health, and can therefore jeopardize the multiple uses of the reservoirs. They reveal the urgent necessity for remedial action, particularly to slow down and to prevent eutrophication.
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Detrimental impacts of toxic Microcystis aeruginosa from Vietnam on life history traits of Daphnia magna / Ảnh hưởng tiêu cực của loài Microcystis aeruginosa có độc ở Việt Nam lên các đặc điểm vòng đời của Daphnia magnaVo, Thi My Chi, Pham, Thanh Luu, Dao, Thanh Son 24 August 2017 (has links) (PDF)
In this study, we tested the long-term and negative effects of microcystin-producing cyanobacterium Microcystis aeruginosa from Vietnam on Daphnia magna under the laboratory conditions. The test organisms were fed with mixtures of green alga Scenedesmus armatus. and toxic M. aeruginosa at different ratios (10% Microcystis + 90% Scenedesmus, 50% Microcystis + 50% Scenedesmus, 100% Microcystis, and 100% Scenedesmus) for over a period of 21 days. The life history traits of the organisms such as, survival, maturation, fecundity were daily recorded. Besides, the intrinsic population rate of D. magna in each treatment was also calculated based on the survivorship, the reproductive age and the clutch size of the animals. The results showed that survival, maturation and reproduction of the D. magna fed with 10, 50 and 100% M. aeruginosa was impaired. Additionally, the intrinsic population rate of the exposed D. magna was lower than that of the control. This study evidenced the adverse effects of toxic M. aeruginosa on both the individual and intrinsic population levels of D. magna. To our knowledge, this is the first report on the chronically detrimental impacts of toxic M. aeruginosa isolated from Vietnam on D. magna and contributed the scientific information on the severe influences of toxic cyanobacteria world wide. / Trong bài viết này, chúng tôi nghiên cứu ảnh hưởng xấu mãn tính của loài vi khuẩn lam Microcystis aeruginosa có khả năng sản sinh độc tố microcysin từ Việt Nam lên Daphnia magna trong điều kiện phòng thí nghiệm. Sinh vật thí nghiệm được cho ăn với hỗn hợp tảo lục Scenedesmus armatus và M. aeruginosa có độc ở các tỷ lệ khác nhau (10% Microcystis + 90% Scenedesmus, 50% Microcystis + 50% Scenedesmus, 100% Microcystis, và 100% Scenedesmus) trong thời gian 21 ngày. Các đặc điểm vòng đời của sinh vật bao gồm sức sống, sự thành thục, sức sinh sản được theo dõi hàng ngày. Bên cạnh đó, tỷ lệ phát triển quần thể của D. magna trong từng lô thí nghiệm cũng được tính toán dựa vào sức sống, tuổi sinh sản và kích cỡ sinh sản của sinh vật. Kết quả cho thấy, sức sống, tuổi thành thục và sự sinh sản của D. magna cho ăn với 10, 50 và 100% M. aeruginosa bị ảnh hưởng xấu. Bên cạnh đó, tỷ lệ phát triển quần thể của D. magna trong lô phơi nhiễm thấp hơn so với đối chứng. Nghiên cứu này chứng minh ảnh hưởng xấu của M. aeruginosa có độc lên cả hai mức độ cá thể và quần thể của D. magna. Theo hiểu biết của chúng tôi, đây là báo cáo đầu tiên về ảnh hưởng xấu mãn tính của M. aeruginosa có độc phân lập từ Việt Nam lên D. magna and đóng góp thêm thông tin khoa học cho những ảnh hưởng nghiêm trọng của vi khuẩn lam có độc trên khắp thế giới.
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Development of Advanced Capillary Electrophoresis Techniques with UV and Mass Spectrometry Detection for Forensic, Pharmaceutical and Environmental ApplicationsFu, Hanzhuo 01 July 2014 (has links)
Capillary electrophoresis (CE) is a modern analytical technique, which is electrokinetic separation generated by high voltage and taken place inside the small capillaries. In this dissertation, several advanced capillary electrophoresis methods are presented using different approaches of CE and UV and mass spectrometry are utilized as the detection methods.
Capillary electrochromatography (CEC), as one of the CE modes, is a recent developed technique which is a hybrid of capillary electrophoresis and high performance liquid chromatography (HPLC). Capillary electrochromatography exhibits advantages of both techniques. In Chapter 2, monolithic capillary column are fabricated using in situ photoinitiation polymerization method. The column was then applied for the separation of six antidepressant compounds.
Meanwhile, a simple chiral separation method is developed and presented in Chapter 3. Beta cycodextrin was utilized to achieve the goal of chiral separation. Not only twelve cathinone analytes were separated, but also isomers of several analytes were enantiomerically separated. To better understand the molecular information on the analytes, the TOF-MS system was coupled with the CE. A sheath liquid and a partial filling technique (PFT) were employed to reduce the contamination of MS ionization source. Accurate molecular information was obtained.
It is necessary to propose, develop, and optimize new techniques that are suitable for trace-level analysis of samples in forensic, pharmaceutical, and environmental applications. Capillary electrophoresis (CE) was selected for this task, as it requires lower amounts of samples, it simplifies sample preparation, and it has the flexibility to perform separations of neutral and charged molecules as well as enantiomers.
Overall, the study demonstrates the versatility of capillary electrophoresis methods in forensic, pharmaceutical, and environmental applications.
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Fate and Persistence of Microcystin Congeners in Lakes and Lake SedimentsZastepa, Arthur January 2014 (has links)
Cyanobacterial blooms and their toxins are a major water quality and potential health risk around the world. This thesis developed an analytical method for microcystin congeners in sediments in order to examine their fate in lakes and establish the history of toxin-producing cyanobacteria in relation to environmental change using lake sediments. A novel method for both intra- and extracellular microcystins in lake sediments was developed, consisting of accelerated solvent extraction, hydrophilic-lipophilic balance solid phase extraction and multiple reaction monitoring-based HPLC-MS/MS quantitation. The method achieved comparable recoveries of intra- and extracellular cyanotoxins based on nine microcystins and nodularin (marine analogue). The analytical method was validated using surficial and deeper sediments from seven lakes of diverse geography and trophic state. To study the fate of microcystins, a multi-year, whole lake study of Microcystis blooms was conducted to obtain both in situ and in vitro half-life estimates of microcystin-LA (MC-LA), an understudied, but increasingly reported microcystin. MC-LA appeared to undergo slower rates of decomposition and persist longer than the more frequently studied MC-LR. Experimentally, high light intensity increased in vitro decomposition of dissolved MC-LA while high temperature enhanced decomposition in the particulate phase. Sediment deposition measurements and estimates of sediment-pore water distribution coefficients, sediment accumulation rates, and diffusive fluxes indicated that microcystin congeners differ in their fate. Notably, MC-LA preferentially distributed into pore water and remobilized (by diffusion) from sediments and into overlying water while MC-RR adsorbed more strongly to sediment particles. Finally, the sediment record of an eutrophic lake of major recreational importance was examined to identify possible drivers of toxigenic cyanobacteria and determine if the perceived increase in toxigenic cyanobacteria could be corroborated. Microcystins were detected to the bottom of the core (early 1800s), indicating that toxigenic cyanobacteria were present prior to the first permanent settlements. Microcystins were significantly correlated with changes in diatom-inferred nutrients (DI-TP and DI-TKN) within the sediment core as well as with specific algal pigments. Sediment microcystins in the upper layers also significantly correlated with a 20-year monitoring record for water column microcystins suggesting that sediment microcystins can be used as a proxy for past surface water conditions.
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Analyse des cyanotoxines dans différents organismes aquatiques et habitats de la réserve écologique de la Rivière-aux-BrochetsSkafi, Mourad 04 1900 (has links)
La diversité et la distribution des cyanobactéries dans les écosystèmes aquatiques conduisent à des effets nuisibles dans l’eau par la production d’une variété de toxines cyanobactériennes qui présentent des risques pour la faune et la santé humaine. Différentes techniques analytiques émergentes ont été développées pour détecter et quantifier les toxines cyanobactériennes dans l'environnement. Dans ce mémoire nous avons examiné la présence de cyanotoxines multi-classes, dont 12 microcystines, les anatoxines, la cylindrospermopsine (CYN), les anabaenopeptines (AP-A, AP-B) et la cyanopeptoline-A dans les eaux de surface et les poissons sauvages. L'échantillonnage a été conduit pendant l’été 2018, dans l'écosystème fluvial de la réserve écologique de la Rivière aux Brochets (QC, Canada) près de la Baie Missisquoi (Lac Champlain). La méthode analytique employée combine la chromatographie liquide ultra haute performance et une ionisation par électronébuliseur (UHPLC-ESI) avec l’usage d’un spectromètre de masse triple quadripôle. Sur les 18 cyanotoxines ciblées, 14 ont été détectées dans des échantillons d'eau de surface impactés par la floraison ; les toxines ont culminé au début de la mi-septembre avec les concentrations les plus élevées de MC-LR (3,8 μg L-1) et MC-RR (2,9 μg L-1). Parmi les 71 poissons prélevés sur le terrain (10 espèces au total), 38% avaient des détections positives d'au moins une cyanotoxine. Dans les échantillons positifs, les plages de concentration dans le muscle du poisson étaient les suivantes : la somme des microcystines ΣMC (0,16-9,2 μg kg-1), la CYN (46-75 μg kg-1), et les anabaénopeptines AP-A (1,1-5,4 μg kg-1) et AP-B (0,01 à 5,0 μg kg-1). Dans l'ensemble, 17% des échantillons de poisson étaient positifs pour AP-A ou AP-B. A notre connaissance, ceci constitue le premier signalement de bioaccumulation d'anabaénopeptines dans la faune. La somme maximale des concentractions des microcystines ΣMC dans les poissons était 1,15 fois plus élevées que la recommandation de l'apport quotidien (8 μg kg-1 de tissu-1) de l'Organisation mondiale de la santé (OMS) pour les adultes et équivalaient presque à la valeur dérivée pour les jeunes enfants 9.3 μg kg-1. La concentration de CYN était également environ 3 fois plus élevée que la limite dérivée des valeurs recommandées pour la santé humaine. / The diversity and widespread distribution of cyanobacteria in aquatic ecosystems lead to harmful effects in water through the production of a variety of cyanobacterial toxins, which pose a great danger to fauna and human health. Different emerging analytical techniques have been developed to detect and quantify cyanobacterial toxins in the environment. In this thesis we examined the presence of multi-class cyanotoxins, including 12 microcystins, anatoxins, cylindrospermopsin (CYN), anabaenopeptins (AP-A, AP-B) and cyanopeptolin-A in surface water and wild fish. Sampling was conducted during the 2018 summer season in the fluvial ecosystem of the Pike River ecological reserve (QC, Canada) near Missisquoi Bay, Lake Champlain. This study was carried out using an analytical method combining ultra-high-performance liquid chromatography and ionization by electrospray (UHPLC-ESI) with the use of a triple quadrupole mass spectrometer. Of the 18 cyanotoxins targeted, 14 were detected in surface water samples impacted by the bloom; toxins peaked in early mid-September with the highest concentrations of MC-LR (3.8 μg L-1) and MC-RR (2.9 μg L-1). Among the 71 fish sampled in the field from 10 species, 38% had positive detections of at least one cyanotoxin. In positive samples, the concentration ranges in fish muscle were as follows: the sum of microcystins ΣMC (0.16-9.2 μg kg-1), CYN (46-75 μg kg-1), AP -A (1.1-5.4 μg kg-1) and AP-B (0.01 to 5.0 μg kg-1). Overall, 17% of the fish samples were positive for AP-A or AP-B; to our knowledge, this is the first report of accumulation of anabaenopeptins in wildlife. The maximum sum ΣMC of microcystin concentrations in fish was 1.15 times higher than the recommendation of the World Health Organization (WHO) Daily Intake (8 μg kg-1 tissue-1) for adults and was almost equivalent to the derived value for young children 9.3 μg kg-1. The concentration of CYN was also approximately 3 times higher than the limit derived from the recommended human health values.
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