Whether active or passive, dispersal accompanied by gene flow shapes the population genetics and evolutionary divergence of species. Indirect methods which use genetic markers have the ability to assess effective dispersal—that which resulted in gene flow. My objective was to see if an aquatic insect and an obligate aquatic invertebrate show similar phylogeographic patterns and genetic uniqueness. Hyalella azteca and Callibaetis americanus were collected from 4-5 springs in each of six basins in the Great Basin of western North America. No dispersal or genetic studies of C. americanus have been conducted to date. However, several studies focusing on mtDNA diversity of H. azteca have revealed a tremendous degree of cryptic diversity in the desert springs of the Great Basin. Nested clade phylogeographical analysis (NCPA), FST values, AMOVA, and Mantel tests were used to examine geographical associations. I also used traditional phylogenetic approaches including maximum parsimony (MP) and likelihood (ML) analyses using cytochrome c oxidase subunit I (COI), 28S, and 16S as genetic markers. The mitochondrial COI sequence divergences in C. americanus were higher than H. azteca COI divergences within springs but lower among springs. FST values were very high in H. azteca reaching near fixation for certain alleles. C. americanus FST values were lower suggesting greater gene flow and, consequently, greater dispersal rates. Even though Mantel tests did not detect significant isolation by distance when evaluating all haplotypes together, nested clade analysis was able to examine smaller networks of related haplotypes and detect significant isolation by distance. Whereas the genetic structure in C. americanus was dominated by restricted gene flow with isolation by distance, H. azteca was characterized more by gradual range expansion followed by fragmentation. Mayflies likely showed more gene flow than amphipods because of their flight capabilities, but movement was still restricted by long distances between isolated springs.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-01-16T17:06:20Z No. of bitstreams: 1 ThiagoFariasNobrega_DISSERT.pdf: 1767927 bytes, checksum: 4bc39921c977a9668ed14d461e4bc936 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-01-19T14:01:37Z (GMT) No. of bitstreams: 1 ThiagoFariasNobrega_DISSERT.pdf: 1767927 bytes, checksum: 4bc39921c977a9668ed14d461e4bc936 (MD5) / Made available in DSpace on 2018-01-19T14:01:37Z (GMT). No. of bitstreams: 1 ThiagoFariasNobrega_DISSERT.pdf: 1767927 bytes, checksum: 4bc39921c977a9668ed14d461e4bc936 (MD5) Previous issue date: 2015-02-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Os efeitos delet?rios das atividades humanas ao meio ambiente s?o temas de v?rios estudos e consequentemente de mudan?as de paradigmas. Exemplo disso, ? que at? pouco tempo acreditava-se em um infinito poder de dilui??o dos ecossistemas em rela??o aos agentes t?xicos, e pouco se conhecia dos efeitos dos contaminantes aos organismos. Em ?reas de intenso crescimento urbano ? comum o descarte indiscriminado de subst?ncias t?xicas e consequentemente o desequil?brio das din?micas naturais dos ecossistemas. Dentre estes, os lagos s?o complexos e importantes fontes de ?gua superficial, abrigam esp?cies e t?m influ?ncia sobre o clima da regi?o ao seu redor. A Lagoa de Extremoz/RN, litoral Nordeste do Brasil, ? um corpo h?drico localizado numa regi?o de r?pido crescimento urbano. Est? sob influ?ncia de atividades rurais, urbanas e industriais, al?m do regime clim?tico prop?cio a per?odos prolongados de estiagem. O abastecimento de cerca de 300.000 pessoas depende da estabilidade h?drica dessa lagoa. O objetivo deste trabalho foi descrever as rela??es entre par?metros f?sicos, componentes qu?micos e as respostas observadas nos ensaios ecotoxicol?gicos, buscando informa??es sobre as varia??es sazonais. Para isso, foram realizadas, em dois anos, 2013 e 2014, an?lises f?sicas, qu?micas e ecotoxicol?gicas da ?gua e do sedimento superficial, em duas esta??es de amostragem na Lagoa de Extremoz (T1 e T2) e uma no rio Guajiru (T3). Nas amostras de ?gua foram aferidos o pH, turbidez, cloreto, OD, nitrato, nitrito, fosfato reativo total, metais e dureza. Para avaliar a toxicidade das amostras foram usados Ceriodaphnia dubia e C. silvestrii como organismos-teste. No sedimento foram aferidos os teores de carbonato, mat?ria org?nica e metais, caracteriza??o granulom?trica e avaliado os efeitos t?xicos sobre a sobreviv?ncia de Hyalella azteca. Verificou-se no per?odo de estiagem, ocorr?ncia de toxicidade aguda aos Hyalella azteca e nesse mesmo per?odo as concentra??es de metais em ?gua e sedimento tamb?m foram maiores. Em uma an?lise espacial percebeu-se que a maioria das amostras com efeito t?xico aos organismos-teste e com maiores concentra??es de metais foram coletadas na esta??o T2. Portanto, existem evid?ncias de que a qualidade da ?gua da Lagoa de Extremoz esteja diminuindo gradualmente e este processo se agrava pontualmente nos per?odos de menor pluviometria e na esta??o de amostragem T2. / The deleterious effects of human activities on the environment, are subject of several studies and therefore paradigm shifts. Until recently thought up in an infinite power dilution of ecosystems in relation to toxic agents, and little was known of the effects of contaminants to organisms. The intense urban growth contributes to the increase of toxic substances and the instability of the environment dynamics. Among these, the lakes are complex ecosystems important for sustaining life, by stock and provide water, to have influence on microclimates and others. The Extremoz Lake/RN, northeastern Brazil, is a water body located in a fast urban growth area. It is under the influence of rural, urban and industrial activities, further on the climate regime conducive to prolonged periods of drought. The supply of approximately 300.000 people depended on the hydraulic stability of this lake The aim of this study was to describe the relationship between physical, chemical components and the responses observed in the ecotoxicological tests, seeking information on seasonal variations. Physical, chemical and ecotoxicological analysis of water and surface sediment were held in two years, 2013 and 2014, in two sampling stations in Extremoz Lake (T1 and T2) and in Guajiru (T3) river. In all the water samples were measured pH, turbidity, chloride, DO, nitrate, nitrite, the total reactive phosphate, hardness and metals. Ceriodaphnia dubia and C. silvestrii as test organisms were used to evaluate the toxicity of the water samples. In the sediment were measured the carbonate, organic matter, metals, particle size, and evaluated the toxic effects on the survival of Hyalella Azteca. In dry season occurred acute toxicity to Hyalella azteca and in the same period the concentrations of metals in water and sediment were also higher. In a spatial analysis, it was noticed that most of the samples with toxic effects on test organisms and with the highest metal concentrations were collected at T2. So there is evidence that the Extremoz Lake quality is gradually declining and this process is occasionally worsens during periods of lower rainfall and sampling station T2.

Laboratory single-species toxicity tests are used to assess the effects of contaminants on aquatic biota. Questions remain as to how accurately these controlled toxicity tests predict sitespecific bioavailability and effects of metals. Concurrent 42-day Hyalella azteca exposures were performed with cadmium and final treated municipal effluent in the laboratory and at the University of North Texas Stream Research Facility. Further laboratory testing in reconstituted hard water was also conducted. Endpoints evaluated include survival, growth, reproduction, and Cd body burden. My results demonstrate that laboratory toxicity tests may overestimate toxicity responses to cadmium when compared to effluent dominated stream exposures. Discrepancies between endpoints in the three tests likely resulted from increased food sources and decreased cadmium bioavailability in stream mesocosms

Silver nanoparticles (AgNPs) are known to cause ecotoxic effects, but there are no existing derived ambient water quality criteria (AWQC) for these nanomaterials to protect freshwater aquatic life due to insufficient toxicological data. We exposed Hyalella azteca to silver nitrate, citrate-coated AgNPs (citrate-AgNPs), and polyvinylpyrrolidone-coated AgNPs (PVP-AgNPs) in a 10-day and 28-day water-only static renewal system with clean sand as a substrate for the amphipods and compared their point estimates with the United States Environmental Protection Agency (USEPA) AWQC for silver. We observed that all treatments decreased the survival, growth, and biomass of H. azteca, and the order of toxicity was AgNO > citrate-AgNPs > PVP-AgNPs. The LC50s of AgNO, citrate-AgNPs, and PVP-AgNPs were 3.0, 9.6, and 296.0 µg total Ag L, respectively, for the acute exposure and 2.4, 3.2, and 61.4 µg total Ag L, respectively, for the chronic exposure. Acute and chronic EC20s of citrate-AgNPs ranged from 0.5 to 3.5 µg total Ag L while that of PVP-AgNPs ranged from 31.2 to 175 µg total Ag L for growth and biomass. Both Ag released from AgNPs and the nanoparticles contributed to the observed toxicity. The dissolution and toxicity of AgNPs were influenced by surface coating agents, particle size, and surface charge. Most point estimates for AgNPs were above AWQC for silver (4.1 µg L) and the lowest concentration (0.12 µg/L) at which Ag is expected to cause chronic adverse effects to freshwater aquatic life. Our study demonstrates that the current AWQC for silver, in general, is protective of freshwater aquatic life against AgNPs tested in the present study.

A literature review of metal mixture interaction analyses identified that there was not a consistent method to determine the impact of metal mixtures on an aquatic organism. The review also revealed that a majority of the research on mixtures made use of water concentrations only. Therefore research was conducted to determine the relationship between exposure, bioaccumulation and chronic effects of the four elements As, Co, Cr and Mn individually. Mechanistically based saturation models of bioaccumulation and toxicity were determined for the benthic invertebrate Hyalella azteca, from which lethal water concentrations and body concentrations were also determined. These models were then combined with those previously done for the metals Cd, Cu, Ni, Pb, Tl and Zn to model the impact of 10 metal mixtures on bioaccumulation in short term (1-week) exposures and on bioaccumulation and toxicity in chronic (4-week) exposures at “equi-toxic” concentrations. Interactions between the metals were identified in which; Cd, Co and Ni bioaccumulations were significantly inhibited, Tl and Zn bioaccumulations were marginally inhibited, there was no impact on Cr, Cu or Mn bioaccumulation, and both As and Pb bioaccumulation were enhanced by some mixtures of metals. It was determined that strict competitive inhibition may be a plausible mechanism of interaction affecting Co, Cd and Ni bioaccumulation but not for any of the other metals. However, it is possible that other interactions such as non-competitive or anti-competitive inhibition may have been responsible. A metal effects addition model (MEAM) was developed for Hyalella azteca based on both the bioaccumulation (body concentrations) to effects and the exposure (water concentration) to effects relationships developed from the single metal only studies The MEAM was used to predict the impact of metal mixture exposures on mortality. Toxicity was under-estimated when based on measured water or body concentrations, however, its best prediction was based on body concentrations. The MEAM, when based on measured body concentrations, takes bioavailability into account, which is important since the chemical characteristics of water can greatly alter the bioavailability and therefore toxicity of metals. The MEAM was compared to the traditional Concentration Addition Model (CAM), which calculates toxic units based on water concentrations and LC50s or body concentrations and LBC50s. The CAM overestimated toxicity, but had its best prediction when based on water concentrations. Over all, the best fit to observed mortality was the prediction by the MEAM, based on body concentrations. The measurement of bioaccumulated metals and the use of the MEAM could be important in field site assessments since it takes into account changes in bioavailability due to different site water chemistries whereas the traditional CAM based on water concentration does not.

A literature review of metal mixture interaction analyses identified that there was not a consistent method to determine the impact of metal mixtures on an aquatic organism. The review also revealed that a majority of the research on mixtures made use of water concentrations only. Therefore research was conducted to determine the relationship between exposure, bioaccumulation and chronic effects of the four elements As, Co, Cr and Mn individually. Mechanistically based saturation models of bioaccumulation and toxicity were determined for the benthic invertebrate Hyalella azteca, from which lethal water concentrations and body concentrations were also determined. These models were then combined with those previously done for the metals Cd, Cu, Ni, Pb, Tl and Zn to model the impact of 10 metal mixtures on bioaccumulation in short term (1-week) exposures and on bioaccumulation and toxicity in chronic (4-week) exposures at “equi-toxic” concentrations. Interactions between the metals were identified in which; Cd, Co and Ni bioaccumulations were significantly inhibited, Tl and Zn bioaccumulations were marginally inhibited, there was no impact on Cr, Cu or Mn bioaccumulation, and both As and Pb bioaccumulation were enhanced by some mixtures of metals. It was determined that strict competitive inhibition may be a plausible mechanism of interaction affecting Co, Cd and Ni bioaccumulation but not for any of the other metals. However, it is possible that other interactions such as non-competitive or anti-competitive inhibition may have been responsible. A metal effects addition model (MEAM) was developed for Hyalella azteca based on both the bioaccumulation (body concentrations) to effects and the exposure (water concentration) to effects relationships developed from the single metal only studies The MEAM was used to predict the impact of metal mixture exposures on mortality. Toxicity was under-estimated when based on measured water or body concentrations, however, its best prediction was based on body concentrations. The MEAM, when based on measured body concentrations, takes bioavailability into account, which is important since the chemical characteristics of water can greatly alter the bioavailability and therefore toxicity of metals. The MEAM was compared to the traditional Concentration Addition Model (CAM), which calculates toxic units based on water concentrations and LC50s or body concentrations and LBC50s. The CAM overestimated toxicity, but had its best prediction when based on water concentrations. Over all, the best fit to observed mortality was the prediction by the MEAM, based on body concentrations. The measurement of bioaccumulated metals and the use of the MEAM could be important in field site assessments since it takes into account changes in bioavailability due to different site water chemistries whereas the traditional CAM based on water concentration does not.

Hyalella azteca, an amphipod crustacean, is frequently used in freshwater toxicity tests. Since the mid-1980s, numerous organizations have collected and established cultures of H. azteca originating from localities across North America. However, H. azteca is actually a large cryptic species complex whose members satisfy both the biological and the phylogenetic species concepts. Recently, two publications reported that members of the H. azteca cryptic species complex have different toxicity responses to anions and an insecticide. In this study, four members of the H. azteca species complex were identified with DNA barcoding. The genetic variation among the four clades was consistent with interspecific distances between species. These lineages (clades 1, 3, 6, and 8) were cultured in identical conditions and monitored on a weekly basis to determine two life history traits: adult mortality and juvenile production. The large-bodied clades had significantly better survival and juvenile production compared to the small-bodied clade 3. Clade 6 had very low juvenile production and high mortality; therefore, was not included in this study. Unique culture protocols may be required for each clade to optimize growth, survival, and juvenile production in laboratory conditions. Genetic barcoding has identified only two clades in a survey of 17 laboratories. Therefore these two clades (1 and 8) were compared after exposure to copper and nickel 14-day toxicity tests. Clade 8 was 2.3-2.6 times more tolerant to copper exposure than clade 1 based on their LC50 and LC25. Similarly, clade 8 was more tolerant to nickel exposure than clade 1: LC50 was 1.8 times higher for the former. Nickel LBC50 and LBC25 were significantly different between clades by a factor of 2.1-2-8. Mortality (relative to copper concentrations in tissue), growth, and bioaccumulation responses were not significantly different based on overlapping confidence intervals. Although clades 1 and 8 are both large-bodied ecomorphs, these lineages had significantly different body mass (i.e., dry weight) after 14 days. The results of this study indicate that genetically characterized cultures of H. azteca should be used in toxicity tests.

L’eutrophisation et la contamination des sédiments (ex : métaux, nutriments, etc.) sont des problèmes majeurs pour la protection des écosystèmes aquatiques et la protection des ressources d’eau douce. Le lac Saint-Augustin (Québec, Canada) est aux prises avec ces deux problèmes. Depuis 2001, plusieurs épisodes de fleurs d’eau de cyanobactéries sont rapportés durant la saison estivale (P > 70 µg/L). Galvez et collaborateurs ont proposé une méthode de restauration pour ce lac consistant : (I) à réduire la teneur en phosphore de l’eau du lac par coagulation/floculation avec du sulfate d’aluminium et (II) à effectuer un recouvrement actif de roche calcaire et de sable afin d’isoler le floc d’Al, le P et les autres contaminants (ex., métaux) et permettre la recolonisation par le benthos. L’objectif des présents travaux était d’évaluer l’écocompatibilité de cette méthode. Deux approches écotoxicologiques ont été employées : (I) essais de toxicité monospécifiques (croissance de l’algue Pseudokirchneriella subcapitata et de la lentille d’eau Lemna minor; survie et reproduction de la daphnie Daphnia magna et de l’amphipode Hyalella azteca, survie, croissance et émergence de l’invertébré Chironomus riparius ainsi que survie des larves de tête-de-boule Pimephales promelas et des alevins de truites arc-en-ciel Oncorhynchus mykiss) et (II) essai en microcosmes de 2 litres tel qu’adapté par Triffault-Bouchet. La toxicité de l’aluminium et l’impact de la roche calcaire ont été étudiés séparément : d’abord avec eau et sédiments artificiels et ensuite avec eau et sédiments du lac Saint-Augustin. Les résultats démontrent que les algues et les daphnies sont affectées par l’application d’alun. Aucun effet n’a été observé chez les autres organismes. Ceux-ci ont eu la capacité de recoloniser les milieux traités avec l’alun ou avec la technique complète. Ces résultats tendent à démontrer que la méthode de restauration est prometteuse. L’écocompatibilité de la technique ne pourra cependant être démontrée que par des essais in situ et après un suivi étroit du phytoplancton et du zooplancton dans le lac. Toutefois, dans le cadre des présents travaux, l’efficacité des traitements (en terme de déphosphatation) n’a pas permis d’atteindre le critère de qualité du MDDEP (20 µg/L). Ces paramètres devront être étroitement suivis lors des essais in situ. Mots clés : Eutrophisation, sédiments contaminés, restauration de lac, aluminium, recouvrement actif, évaluation écotoxicologique, bioessais monospécifique, microcosmes. / Eutrophication and contaminated sediments (e.g.: nutrients, metals, etc.) are preoccupying issues in aquatic ecosystems and water resource protection. Saint-Augustin Lake (Quebec, Canada) is affected by both problems in addition to the presence of de-icing salts. This small shallow lake has low renewal time and receives an intensive phosphorus transfer from bottom sediments to the water column. Since 2001, the lake presents cyanobacteria blooms every summer (P > 70 μg/L). Galvez and collaborators has proposed a remediation method for this lake that consists of (i) the precipitation of phosphorus present in the water column by the addition of alum, in association with the deposition of (ii) an active capping layer (limestone + sand) to isolated P, Al and other contaminants (e.g., metals) within bottom sediments and allow recolonisation by benthos. The objective of this study is to assess the ecotoxicological impact of that remediation method. Two ecotoxicological approaches are used to assess the impact of this method: (I) single-species bioassays (algae Pseudokirchneriella subcapitata and duckweed Lemna minor growth; daphnid Daphnia magna and amphipod Hyalella azteca survival and reproduction, invertebrate Chironomes riparius survival, growth and emergence, fathead minnow P. promelas and rainbow trouts O. mykiss survival) and (II) a 2-L microcosm as adapted by Triffault-Bouchet. Alum toxicity and limestone impact were studied separately in laboratory: first with artificial water and sediments and then using actual lake overlying water and sediments. The results show that algae and daphnids are sensitive to alum treatment. No significant effects were observed on the other organisms. These organisms had the ability to recolonize the treated media either with alum treatment or with complete treatment. These results suggest that the restoration method is promising. The environmental compatibility of the technique, would be demonstrated after testing in situ with a close monitoring of phytoplankton and zooplankton in the lake. However, the results of this study showed that the treatments dephosphatation efficacity was not enought to reach the MDDEP quality criteria (20 µg/L). Also, the concentration of residual aluminum after treatments was still over the MDDEP quality criteria after treatments. Keywords : Eutrophication, contaminated sédiments, lake restauration, aluminum, active capping, écotoxicological évaluation, monospécific bioessais, microcosms.