Orientador: Gilson Luiz Volpato / Banca: Eliane Gonçalves-de-Freitas / Banca: Mário Luis Orsi / Banca: Leonardo José Gil Barcellos / Banca: Percília Caedoso Giaquinto / Resumo: Atualmente, a função das depressões que machos de muitas espécies de ciclídeos escavam no substrato é para atração de fêmeas para a reprodução. Assim, tais depressões têm sido consideradas um fenótipo estendido dos machos, ou seja, representam um caracter a mais, pelo qual a fêmea pode escolher o melhor macho. Assim, é esperado que somente os machos escavem e, sobretudo, em contextos reprodutivos. No entanto, em nosso estudo 1 vimos que tanto machos quanto fêmeas sexualmente imaturos já escavam essas depressões e que esse comportamento surge também na ausência da interação com coespecíficos e num período restrito da ontogenia. Isso ocorreu tanto para a domesticada tilápia-do-Nilo (Oreochromis niloticus) quanto para o selvagem cará (Geophagus brasiliensis). Isso sugere que o comportamento de escavar tenha uma forte determinação genética e outra função biológica relevante além da reprodução. No estudo 2, investigamos na tilápía-do-Nilo se essa outra função poderia ser a busca por alimento, uma vez que os padrões motores na escavação e no forrageamento do substrato são semelhantes. Contudo, vimos que a resposta de peixes com fome não satisfez as predições dessa hipótese. A fome do peixe não influenciou a frequência com que machos e fêmeas escavaram e nem o tamanho de suas escavações. Finalmente, no estudo 3 mostramos que as escavações de machos e fêmeas servem como refúgio, pois suas características permitem que os peixes se escondam dentro delas, a presença de uma toca no aquário reduz drasticamente a frequência de escavação, as escavações são posicionadas mais distantes de áreas abertas (perigo) e os peixes ficam dentro das escavações durante período de inatividade noturna. Isso indica que as escavações podem ter função de abrigo para evitar perigos potenciais / Abstract: Not available / Doutor

Orientador: Margarida Maria Barros / Resumo: Visando o preparo da tilápia do Nilo, Oreochromis niloticus, para o período de inverno, foram avaliados durante 112 dias diferentes níveis de suplementação de lipídeo e vitamina C, no desempenho produtivo e parâmetros fisiológicos. Foi utilizado um lote de 192 alevinos, com peso médio de 5,57 0,50 g distribuídos aleatoriamente em 32 tanques-rede de 200 L cada, dispostos em oito aquários de 1000 L, dotados de sistema de aquecimento e de recirculação de água com filtragem mecânica e biológica. O experimento foi conduzido num esquema fatorial 2 x 3, com dois níveis de lipídeo (8,0 e 12,0%) e três níveis de vitamina C (300,0; 600,0 e 1200,0 mg de vitamina C/kg da dieta), com quatro repetições, mais dois tratamentos, um ausente de suplementação dos nutrientes testes e outro acrescido de 6,0% de lipídeo e 125,0 mg de vitamina C/kg da dieta. Foram confeccionadas oito rações práticas, isoprotéicas com 32,0% de proteína digestível. O suplemento vitamínico e mineral utilizado foi isento de vitamina C, sendo considerada como fonte somente o adicionado à dieta. A vitamina C utilizada foi a polifosfatada com 35,0% de atividade e a fonte de lipídeo o óleo de soja. Os resultados permitiram concluir que a energia adicional proveniente do lipídeo é depositada na cavidade abdominal na forma de gordura visceral; que a deposição desta gordura como reserva de energia para transpor o período de inverno foi obtida em todos os níveis de suplementação de lipídeo, sendo a suplementação com 8,0% a mais adequada; que a concentração hepática de vitamina C é proporcional a concentração desta na dieta porém, em função da capacidade de reserva do fígado a quantidade de 600,0 mg de vitamina C/kg da dieta mostra-se mais apropriada economicamente e que a ausência de vitamina C prejudica a eritropoiese e a síntese de colágeno. / Abstract: It were evaluated during 112 days different lipid and vitamin C supplementation in practical diets, on growth performance and physiologic parameters of Nile tilapia, aiming the animals preparation for winter time. It was utilized 192 fingerlings, selected from the same spawn, with average weight of 5.57 0.50 g. These were randomly stocked into 32 200-l aquaria, supplied with heat system and recycled water with mechanic and biological filter. The experiment was conducted in factorial design 2 x 3, two levels of lipid (8.0 and 12.0%) and three levels of vitamin C (300.0; 600.0 and 1,200.0 mg of vitamin C/kg diet), with four replicates each treatment, plus two treatments, absence of test nutrients supplementation and supplemented with 6.0% of lipid and 125.0 mg of vitamin C/kg diet. Eight experimental practical diets were formulated to contain 32.0% of digestible protein. Vitamin and mineral mix was absent of vitamin C, considering as a source only the supplemented in diets. The vitamin C utilized was polyphosphate 35.0% activity. Based on the results it was concluded that the additional energy coming from lipid is accumulated in abdominal cavity as visceral fat; fat deposition as a energy reserve for winter time was obtained in all lipid supplementation, however 8.0% supplementation seems to be more appropriate; vitamin C liver concentration is proportional to diet concentration, but because of liver reserve capacity the amount of 600.0 mg of vitamin C/kg diet appears to be more economic appropriate and that vitamin C absence impairment erythropoiesis and collagen syntheses. / Mestre

A Grande São Paulo, composta por 39 municípios, apresenta crescimento urbano e industrial expoente que contribuem para o constante acúmulo de poluentes no ambiente. Ainda assim, exibe áreas verdes remanescentes que contribuem para a sustentabilidade ambiental da cidade, como por exemplo, o Parque Ibirapuera e o Parque Ecológico do Tietê. Nesse contexto, o presente estudo visa verificar a viabilidade em utilizar as O. niloticus como bioindicadoras da poluição por mercúrio no ecossistema aquático dos dois parques da Grande São Paulo. Obteram-se para as tilápias do Nilo do Parque Ibirapuera pertencentes aos pontos de coleta A, B e C durante o inverno as respectivas concentrações de mercúrio total no sangue 2,72±4,20µg/L, 2,45±1,90µg/L e 1,19±0,05µg/L e, no hepatopâncreas as respectivas concentrações de mercúrio de 0,07±0,02 µg/g, 0,06 ±0,005 µg/g e 0,16 ±0,07 µg/g. No verão as tilápias do Parque Ibirapuera apresentaram nos mesmos pontos de coleta as respectivas concentrações de mercúrio total no sangue 2,50±0,84µg/L, 2,67±1,03µg/L e 2,430±0,790µg/L e, no hepatopâncreas as respectivas concentrações de mercúrio de 0,08 ±0,03 µg/L, 0,16 ±0,10 µg/L e 0,09 ±0,03 µg/L. Os peixes do Parque Ecológico do Tietê pertencentes ao ponto de coleta B durante o inverno e verão apresentaram no hepatopâncreas as concentrações de mercúrio total de 0,08±0,01µg/g e 0,10±0,0µg/g e, no sangue as concentrações e mercúrio de 3,59 µg/L e 2,00±1,00 µg/L. A análise estatística de regressão linear foi negativa, alta e com significância estatística apenas entre a idade estimada e a concentração do mercúrio no sangue sugerindo que as tilápias jovens são mais susceptíveis ao acúmulo de mercúrio. Viabilizou-se através deste estudo o uso das tilápias do Nilo como bioindicadores para a poluição por mercúrio nos lagos dos parques da Grande São Paulo. / The Greater São Paulo, formed by 39 cities, presents high urban and industrial growth that contribute to environmental pollution accumulation. In contrast, the same area has residual green areas that contributes to the city\'s environmental sustainability, such as Parque Ibirapuera and Parque Ecológico do Tietê. This study intends to verify the feasebility of using O. niloticus as bioindicators of mercury pollution on the aquatic ecosystem of two Greater São Paulo parks. At winter, on Parque Ibirapuera, the collected Nile Tilapia presented 2,72±4,20µg/L, 2,45±1,90µg/L e 1,19±0,05µg/L mercury concentration on blood and 0,07±0,02 µg/g, 0,06 ±0,005µg/g e 0,16 ±0,07 µg/g mercury concentration on liver for collect points A, B and C respectively. At summer, Nile Tilapias collected on the same three points, presented 2,50±0,84µg/L, 2,67±1,03µg/L e 2,430±0,790µg/L mercury concentration on blood and 0,08 ±0,03 µg/L, 0,16 ±0,10 µg/L e 0,09 ±0,03 µg/L on liver. The Parque Ecológico do Tietê\'s fish, from point B, presented 0,08±0,01µg/g mercury concentration on liver and 3,59 µg/L mercury concentration on blood. At summer, the mercury concentrations were 0,10±0,0µg/g on liver and 2,00±1,00 µg/L on blood. The linear regression statistical analisys was negative, high and with statistical significance only between estimated age and blood mercury concentration, indicating that young tilapia are more likely to accumulate mercury. The use of Nile Tilapia as bioindicators for mercury pollution on Greater São Paulo\'s park lakes was made viable through this study.

Doctor of Philosophy / Department of Chemistry / Daniel A. Higgins / This dissertation describes single-molecule tracking (SMT) studies for the quantitative characterization of one-dimensional (1D) solvent-filled surfactant-templated mesoporous silica (STMS) materials and other nanostructured materials, such as double-stranded DNA. SMT permits the simultaneous and quantitative assessment of the nanoscale and microscale morphologies and mass-transport properties of the materials with nanometer-scale spatial resolution. The efficiency and selectivity of catalytic reactions and chemical separations occurring in liquid-filled mesoporous materials are governed by the translational and orientational mobilities and surface interactions of the incorporated reagents and analytes. Polarization dependent SMT results demonstrate that the dye molecules used as probes of materials nanostructure are tightly confined within the one-dimensional (1D) pores of surfactant-templated mesoporous silica films. Spectroscopic single molecule tracking (sSMT) data reveal that the hydrophobic probe dyes are confined within nonpolar regions of the nanomaterials For this dissertation, surfactant templated mesoporous silica films were prepared by the spin coating of acid catalyzed tetramethoxysilane (TMOS)-based silica sols on glass substrates in the presence of Cetyltrimethylammonium bromide (CTAB). Cylindrical CTAB micelles formed during evaporation of the solvent acted as a structure directing template, forming nanometer-sized one-dimensional pores within the silica films. SMT experiments were performed using a wide-field fluorescence microscope that was sufficiently sensitive to allow detection of the fluorescence from individual dye molecules. A series of perylene diimide (PDI) dyes was employed for basic structural characterization of the silica materials. Single molecule fluorescence was recorded in the form of fluorescence videos. These videos revealed the presence of immobile dye molecules, along with those diffusing in one and two dimensions (1D and 2D). The 1D diffusing molecules provided basic evidence for the confinement mass transport of the dye molecules within the silica mesopores. Spectroscopic single molecule tracking (sSMT) studies served as an extension of basic SMT experiments and were employed to determine the location of the molecules. The polarity sensitive dye Nile Red (NR) was employed in these studies. It exhibits 1D diffusion, consistent with its confinement to the cylindrical pores, as was also the case for the PDI dyes. The sSMT data revealed that the majority of NR molecules were found in nonpolar environments having polarities similar to that of n-hexane. Single molecule emission polarization (SMEP) measurements were employed to explore the orientational confinement of the dyes. The results of these experiments demonstrated that the PDI and NR molecules diffuse with their long axes aligned parallel to the long axis of the pores. All of the dyes employed were found to be orientationally confined to ∼1 nm diameter pathways within the pores. The diffusion coefficient for the dyes was also shown to be ∼10^3 -fold smaller than in bulk solution. The results of the NR studies demonstrate that the dye molecules were confined to the hydrophobic cores of the micelles, and provide support for the conclusion that the PDI dyes are similarly confined. These studies afford an enhanced understanding of how nanostructuring of the pore-filling medium in solvent- and surfactant-filled mesoporous materials governs the mass transport and surface interactions of incorporated reagents and analytes. The dependence of molecular confinement on dye charge and structure was also explored in this dissertation. The confined translational and orientational motions of a series of four different PDI dyes diffusing along one dimension (1D) within individual cylindrical silica mesopores were investigated in these studies. Specifically, the motions of cationic and anionic PDI dyes were compared to those of two uncharged PDIs having different alkane tail lengths. All four dyes exhibited populations that were immobile, along with separate populations that diffused in either 1D or 2D. The anionic and cationic PDI dyes exhibited the largest and smallest populations, respectively, of immobile molecules, suggesting that electrostatic interactions between the charged dyes and the cationic surfactant head groups play a significant role in limiting molecular motion. The cationic and anionic PDI dyes also exhibit the largest populations of 2D diffusing molecules, suggesting they may more readily pass between the cylindrical micelles and through the silica pore walls. All four dyes also emit strongly polarized fluorescence as they move in 1D, indicating they are orientationally confined within the nanochannels. Nile Red dye was used to determine the dielectric constant, ε, of nonpolar microenvironments in double-stranded DNA (ds-DNA) single molecules both in aqueous buffer solution and when adsorbed on amine-modified chemical gradient surfaces. The value of ε within the DNA decreased with increasing buffer concentration. Values of ε ∼ 6.75 and ∼3.00 were obtained in 0.1 mM phosphate buffered saline (PBS) and in 10 mM PBS, respectively. Similar effects were observed upon adsorption to chemically graded amine-modified silica surfaces. Under 1 mM buffer, ε was measured to be ∼2.84 and ∼1.90 at the low amine (high silica), and high amine (low silica) ends of the gradient, respectively. An increase in the buffer concentration again led to a decrease in ε, but only at the low amine end. It is concluded that high buffer concentrations and binding to an amine surface cause condensation of the ds-DNA, forming less polar microenvironments within its structure. These results provide important knowledge of the factors governing the polarity of DNA microenvironments to which intercalators bind.

abstract: The Egyptian New Kingdom city of Akhetaten (modern: Tell el-Amarna, el-Amarna, or simply Amarna) provides a unique opportunity to study ancient biocultural dynamics. It was a disembedded capital removed from the major power bases of Memphis and Thebes that was built, occupied, and abandoned within approximately 20 years (c. 1352–1336 BCE). This dissertation used the recently excavated Amarna South Tombs cemetery to test competing models for the development of disembedded capitals, such as the geographic origin of its migrants and its demographic structure in comparison to contrastive models for the establishment of settlements. The degree to which biological relatedness organized the South Tombs cemetery was also explored. The results suggest that the Nile Valley into the New Kingdom (1539–1186 BCE) was very diverse in dental cervical phenotype and thus highly mobile in respects to gene flow, failing to reject that the Amarna city was populated by individuals and families throughout the Nile Valley. In comparison, the Amarna South Tombs cemetery contained the least amount of dental phenotypic diversity, supporting a founder effect due to migration from larger, more diverse gene pools to the city or the very fact that the city and sample only reflect a 20-year interval with little time to accumulate phenotypic variation. Parts of the South Tombs cemetery also appear to be organized by biological affinity, showing consistent and significant spatial autocorrelation with biological distances generated from dental cervical measurements in male, female, and subadult (10–19 years of age) burials closest to the South Tombs. This arrangement mimics the same orderliness in the residential areas of the Amarna city itself with officials surrounded by families that supported their administration. Throughout the cemetery, adult female grave shaft distances predict their biological distances, signaling a nuclear family dynamic that included many females including mothers, widows, and unwed aunts, nieces, and daughters. A sophisticated paleodemographic model using simulated annealing optimization projected the living population of the South Tombs cemetery, which overall conformed to a transplanted community similar to 19th century mill villages of the United States and United Kingdom. / Dissertation/Thesis / Doctoral Dissertation Anthropology 2018

DeKalb and Fulton Counties, which share the metropolitan Atlanta area, have seen an increase in West Nile infected vector mosquitoes; the increase is associated with close proximity to combined sewer overflow facilities. Despite completion of the remediation system in 2008, the mosquito population testing positive for West Nile virus has increased each year from 2010 through 2012. Guided by the Geographical Information System framework and using spatial analysis and regression analyses, this study described and quantified the relationship between sewer system overflows and amplification of vector mosquitoes; an additional goal was to investigate their proximity to human cases of West Nile VIrus (WNV) infections. Comparing the prominence of all WNV vectors revealed how different mosquito species occupy the area. The Culex species was not detected in adult surveillance in 2012; however, the infection rate of mosquito pools increased by 15% and the human infection more than doubled. The influence of sewer system overflows became pronounced when this study analysis also identified that a proportion of West Nile-virus positive mosquito pools was significantly higher in approximately 58% of trap sites within 1 km of sewer overflow events and 30% over 1 km distance from sewer overflow events. Thus, the research contributes to shared information both in support of previous findings and considering novel sources that contribute to the proliferation of WNV. This research can help reduce the rate of WNV infection and decrease the resources needed to protect the public.

West Nile is an important novel virus in the United States, having spread rapidly since it was first detected in New York in 1999. The Centers for Disease Control and Prevention as well as many State Health Departments, have mandated programs for surveillance of West Nile Virus activity. These programs incorporate many different aspects including existing arboserology programs with additional testing for West Nile Virus and new plans that incorporate active and passive surveillance methods. The objective of this study was to examine all aspects of the Florida West Nile surveillance program to determine if there was transmission in the animal systems prior to human cases. The predictive analyses were done using regional data graphs, spatial information, correlations and regression models. Data for sentinel chickens, bird necropsy and mosquito pool surveillance from participating counties in Florida were obtained from the State of Florida surveillance database. The human data was obtained from the State of Florida reportable disease database for each county whether participating in the state surveillance programs or not. Clinical cases were examined by demographics (gender and age) and an incidence rate was calculated to demonstrate the effects of disease. Specific statistical methods used included Pearson's coefficient correlation, Poisson distribution regression modeling to show if any of the surveillance systems were predictors for human disease. The incidence rate analysis for clinical cases showed clustering of cases in adjacent counties within a region where Florida's panhandle and adjacent counties northeast had the highest incidence. Florida's central and southern regions had moderate human incidence. This provides useful information in transmission geography for prevention and control measures. Demographic analysis showed that there were twice as many males than females diagnosed with West Nile in Florida, this was true across the groups as well. The highest number of cases was seen within the age group over 55 years of age for West Nile Neuroinvasive Disease and for West Nile Fever the highest number of cases was within the 36-54 age range. The temporal distribution was determined using graphical representations of all of the surveillance types and clinical cases. In order to include all relevant data, the temporality was set from week 20 to week 52. This study found that all of the surveillance types (dead birds, mosquitoes and sentinels) offered a specialized strength for predicting clinical cases. However, mosquitoes proved to be the least efficient out of the three surveillance systems. The regional and spatial analysis showed that positive dead birds and sentinels provided the coverage for the surveillance systems in the state. However, Pearson's correlation coefficient was low for sentinel surveillance; this may be due to higher participation showing West Nile Virus activity in areas (especially rural) that have no reported human cases. This analysis did show that West Nile is detected in mosquito pool samples before it is detected in the dead bird or sentinel surveillance systems which provides an earlier warning for human cases. The Poisson distribution regression model was only useful for the pooled years and 2003. These showed that mosquitoes, positive dead birds and sentinels were good predictors for clinical cases for the combined years and dead birds and sentinels were significant for 2003 as well. The recommendations based on the results from this study would be to continue all the current surveillance efforts but with the following enhancements: 1. Increase the coverage and consistency of submissions for all surveillance types. 2. Set standard levels of participation for all counties based on the regional analyses and populations at risk. 3. Create standardized approaches for sampling, shipping and submitting samples (especially for mosquito pool submissions) and require that participating counties adhere to these standards. 4. Only submit specific birds known to be especially susceptible to West Nile Virus (e.g. corvids). 5. Targeted prevention and education strategies for higher risk groups based on their potential levels of exposure.

Within the state of Florida, there are three arboviruses of public health importance that can cause neuroinvasive disease in humans: West Nile Virus, Saint Louis Encephalitis Virus, and Eastern Equine Encephalitis Virus. Mosquitoes (Diptera: Culicidae) within the genus Culex are known and suspected vectors of these diseases. The vectors of these diseases can be present in urban wetland habitats that allow for exposure to residential communities. Vector ecology must be investigated in order to understand the dynamics of disease transmission. In Hillsborough County, Florida the spatial and temporal distribution of these vectors are not well established. An ecological study was conducted in the University of South Florida's Ecopreserve using trapping methodologies to sample the adult and gravid females as well as collect the egg population. Collections were made at three spatial points for the duration of July through December 2013 and compared to meteorological variables. Culex erraticus, a proposed bridge vector of Eastern Equine Encephalitis, was the most abundant adult species and gravid female captured. Culex nigripalpus, primary Floridian vector of Saint Louis Encephalitis and bridge vector of West Nile Virus, was the second most abundant adult species caught as well as the majority of eggs collected. Based on the results collected, the presence of Culex erraticus and Culex nigripalpus was confirmed. The majority of Culex erraticus adults were collected in September and October and Culex nigripalpus adults were the highest in July and August. The results of the gravid and egg collection generated crucial insight regarding methodology for studying vector ecology within this urban wetland habitat. However, modeling at spatial points based on meteorological variables yielded inconsistent results that illicit further investigation regarding these arboviral vectors of disease.

Thesis (Ph. D.)--Ohio State University, 2006. / Title from first page of PDF file. Includes bibliographical references (p. 76-86).

Two cellular innate responses, the dsRNA protein kinase (PKR) pathway and the 2'-5' oligoadenylate synthetase (OAS)/RNase L pathway, are activated by dsRNAs produced by viruses and reduce translation of host and viral mRNAs. PKR activation results in eIF2a phosphorylation. As a consequence of eIF2a phosphorylation, stress granules (SGs) are formed by the aggregation of stalled SG proteins with pre-initiation complexes and mRNA. West Nile virus (WNV) infections do not induce eIF2a phosphorylation despite upregulation of PKR mRNA and protein suggesting an active suppression of PKR activation. Assessment of the mechanism of suppression of PKR activation in WNV-infected cells indicated that WNV infections do not induce PKR phosphorylation so that active suppression is not required. In contrast to infections with "natural" strains of WNV, infections with the chimeric W956 infectious clone (IC) virus efficiently induce SGs in infected cells. After two serial passages, the IC virus generated a mutant (IC-P) that does not induce SGs efficiently but does induce the formation of NS3 granules that persist throughout the infection. This mutant was characterized. 2'-5' oligoadenylate synthetases (OAS) are activated by viral dsRNA to produce 2-5A oligos that activate RNase L to digest viral and cellular RNAs. Resistance to flavivirus-induced disease in mice is conferred by the full-length 2'-5' oligoadenylate synthetase 1b (Oas1b) protein. Oas1b is an inactive synthetase that is able to suppress the in vitro synthetase activity of the active synthetase Oas1a. The ability of Oas1b to inhibit Oas1a synthetase activity in vivo and to form a heteromeric complex with Oas1a was investigated. Oas1b suppressed 2-5A production in vivo. Oas1a and Oas1b overexpressed in mammalian cells co-immunoprecipitated indicating the formation of heteromeric complexes by these proteins. Unlike mice, humans encode a single OAS1 gene that generates alternatively spliced transcripts encoding different isoforms. Synthetase activity has previously been reported for only three of the isoforms. The in vitro synthetase activity of additional OAS1 isoforms was analyzed. All tested isoforms synthesized higher order 2-5A oligos. However, p44A only produced 2-5A dimers which inhibit RNase L.