published_or_final_version / abstract / toc / Statistics and Actuarial Science / Master / Master of Philosophy

In addition to their role in hemostasis, platelets appear to contribute to vascular inflammatory diseases. Platelets achieve this through the secretion of various prothrombotic and pro-inflammatory molecules. Platelet secretion is mediated by integral membrane proteins called Soluble NSF Attachment protein REceptors (SNAREs). SNAREs come from both granule/vesicle membranes (v-SNAREs) and target membranes (t-SNAREs) to form a trans-bilayer complex that promotes membrane fusion and subsequent granule cargo release. The work described in this dissertation dissects various, yet related aspects of platelet secretion in both physiological relevant and pathological circumstances. Atherosclerosis is a leading cause of death in the westernized countries and a major contributor to heart attacks and strokes. Given the potential involvement of platelets in atherosclerosis and previous work from our laboratory showing that VAMP-8 is the primary v-SNARE for platelet secretion, one part of this dissertation focuses on the role of VAMP-8- mediated secretion in atherosclerosis. The data showed that the deletion of VAMP-8 in the ApoE-/- null model of chronic atherosclerosis attenuated plaque development compared to the wild type littermates. Aged (50 week) VAMP-8-/-/ApoE-/- mice showed a reduction in lesion size compared to ApoE-/- controls, as measured by Oil Red-O staining of the plaques in the aortic sinus and by en face analysis of plaques in the aortic arch. These data show that the loss of VAMP-8 attenuates the development of atherosclerotic plaques and suggest that platelet secretion contributes to atherosclerosis. Considering the vital role of platelet secretion in both physiological and pathological conditions, it is essential to understand how it is regulated. SNARE proteins are controlled by a range of regulatory molecules that affect where, when, and with whom they form trans-bilayer complexes for membrane fusion. One family of such regulators is the Munc18 family: platelets contain three (Munc18a-c). The second part of this dissertation focuses on the role of Munc18b/STXBP2. Mutations in the Munc18b/STXBP2 gene underlie Familial Hemophagocytic Lymphohistocytosis type 5 (FHL5), which is a life- threatening disease caused by dysregulation of the immune system. Platelets from two biallelic FHL5 patients had almost undetectable levels of Munc18b/STXBP2 levels; the levels of Munc18a increased slightly and Munc18c levels were unaffected. Syntaxin 11 levels were affected but the levels of other secretory machinery proteins were normal. Platelet secretion from dense and alpha granule in two biallelic patients and the one heterozygous patient was decreased. The release of serotonin from dense granules, and platelet factor 4 (PF4) from alpha granules was profoundly affected in the biallelic patients and partially affected in the heterozygote heterozygous patient. Lysosome release was affected only from the platelets of the biallelic patients. These data indicate that Munc18b plays a key role in platelet secretion. Ras is the prototypical member of a family of low molecular weight, GTP-binding proteins. It affects various cellular functions by cycling between an active, guanine triphosphate (GTP) and an inactive guanine diphosphate (GDP) -bound state. Little is known about the role of Ras activation in platelets. The third part of this dissertation focuses on what could be learned about Ras’ role by analyzing platelets from patients with Noonan Syndrome. Specific mutations in the genes encoding elements of Ras signaling pathways are associated with Noonan Syndrome. Platelets from Noonan Syndrome patients with a mutation in kRas (F156V) were analyzed and shown to have a defect in aggregation in response to low levels of agonist. These data suggest that Ras may play a functionally relevant role in platelet activation. In summary, the experiments presented in investigations of this dissertation support a role for platelet secretion in several pathological conditions and suggest that platelet secretion assays may serve as useful as diagnostic tools for some genetic diseases. In addition, these studies elucidate the importance of understanding the regulation of platelet exocytosis, to drive the development of new antithrombotic therapeutics.

Colorectal cancer (CRC) is defined as a cancer appearing in the colon or in the rectum. In Sweden, ~ 6300 individuals were diagnosed with the disease in 2014 and ~ 2550 individuals diagnosed with CRC die each year due to their cancer. Surgery is the main treatment option of CRC and a survival rate of ~ 10 % is estimated if distant metastases have developed. It is therefore of importance to find factors that may be useful together with tumour, node, metastasis (TNM) stage to establish early CRC diagnosis, prognosis and follow-up of CRC patients. The aim of this thesis was to study the possible association of CD93, PLA2G4C, PDGF-D and inflammatory cytokines with CRC disease progression. In a prospective study approach CD93 and PLA2G4C single nucleotide polymorphisms (SNPs) were of potential importance in CRC prognosis. The T/T genotype of CD93 was associated with an increased CD93 expression in CRC tissue. Further, CRC patients carrying this genotype were associated with disseminated CRC at diagnosis and a lower recurrence-free survival after surgery. The A allele of a SNP of PLA2G4C was a stronger predictor for CRC-specific mortality than the conventional risk factors used in the clinic for selection of TNM stage II patients for adjuvant treatment. This indicates that the T/T genotype of CD93 and the A allele of PLA2G4C may be potential genetic factors related to disease severity and spread. Furthermore, they distinguish CRC patients that may benefit from a more comprehensive follow-up and adjuvant treatment. To study the putative involvement of PDGF-D in CRC the effects of PDGF-D signalling was studied in vitro. PDGF-D signalling altered the expression of genes of importance in CRC carcinogenesis and proliferation which was blocked by imatinib, a tyrosine kinase inhibitor. This indicates that PDGF-D signalling may be an important pathway in CRC progression and a potential target in CRC treatment. The analysis of various inflammatory cytokines in plasma at diagnosis showed an association between high levels and increased total- or CRC-specific mortality two years after surgery. High levels of CCL1 and CCL24 was the only cytokines strongly correlated with a worse CRC prognosis after statistical adjustments and may be of interest for further evaluation. In conclusion, this thesis presents circulating and genetic factors such as CD93, PLA2G4C, PDGF-D, CCL1 and CCL24 that may be of importance in CRC progression and may be of clinical value together with TNM stage in establishing prognosis. / Kolorektal cancer är en tumör i kolon eller rektum. I Sverige diagnosticerades år 2014 ca6300 individer med denna cancertyp och ca 2550 personer dör årligen till följd av kolorektalcancer. Operation är det huvudsakliga behandlingsalternativet för kolorektal cancer och vidfjärrmetastaser är överlevnaden < 10 %. Det är därför viktigt att hitta markörer somtillsammans med TNM-stadium kan ge tidig information om sjukdomens prognos och lämpliguppföljning av patienter. Utveckling av kolorektal cancer sker genom ackumulering av genetiska mutationer ochepigenetisk nedreglering av tumörsuppressorgener. Därutöver spelar interaktionen mellantumören och dess närmaste omgivning, innehållande tillväxt- och inflammatoriska faktorer,en viktig roll i tumörens utveckling och metastasering. Syftet med avhandlingen var att studera associationen mellan CD93, PLA2G4C, PDGF-D samtinflammatoriska cytokiner och kolorektal cancer progression. En prospektiv studie visade att CD93 och PLA2G4C SNP var potentiellt viktiga förbedömningav kolorektal cancer prognos. T/T genotypen av SNP rs2749817 i CD93 var associerad medhögre uttryck av CD93 i kolorektal cancer vävnad, främst bland patienter i stadium IV.Därutöver observerades fler återfall efter operation hos patienter med T/T genotypen. Aallelen hos PLA2G4C SNP rs1549637 är en möjligtvis bättre markör för cancerspecifiköverlevnad vid stadium II än faktorer som idag används för att selektera patienter tilladjuvant behandling. Sammantaget antyder detta att T/T genotypen av CD93 och A allelenav PLA2G4C kan vara genetiska markörer relaterade till allvarlig tumörsjukdom ochspridning. Därutöver kan de eventuellt selektera patienter som kräver tätare uppföljning ochadjuvant behandling. För att studera den förmodade inblandningen av PDGF-D i kolorektal cancer undersöktesdess effekt på PDGF-D signalering in vitro. PDGF-D signaleringen förändradegenexpressionen av gener involverade i tumörutveckling och spridning, vilken kundeblockeras av tyrosinkinashämmaren imatinib. Det antyder att PDGF-D signalering kan vara enviktig faktor vid kolorektal cancer progression och ett potentiellt mål för behandling. Analysen av ett flertal inflammatoriska cytokiner visade en korrelation mellan högacytokinnivåer och ökad cancerspecifik och total dödlighet två år efter operation. Höga CCL1och CCL24 nivåer var de enda faktorerna som förblev signifikant associerade medcancerspecifik mortalitet vid fördjupad statistisk analys och bör studeras vidare. Sammanfattningsvis presenterar denna avhandling cirkulerande och genetiska faktorersåsom CD93, PLA2G4C, PDGF-D, CCL1 and CCL24 som eventuellt är viktiga vid bedömning avkolorektal cancer progression tillsammans med TNM stadium.

Epidemiological observations have linked increased host iron with malaria susceptibility. At the same time, blood-stage malaria infection is associated with potentially life-threatening anemia. To improve our understanding of these relationships, this work presents an examination of the mechanisms controlling the upregulation of the hormone hepcidin, the master regulator of iron metabolism, in malaria infection. Chapter 2 presents data from a mouse model of malaria infection which indicate that hepcidin upregulation in malaria infection is associated with increased activity of the sons of mothers against decapentaplegic (Smad) signaling pathway. Although the canonical Smad pathway activators, bone morphogenetic proteins (Bmp) are not increased at the message level following infection, activin B, which has been recently shown to increase hepcidin through the Smad signaling pathway in conditions of inflammation and infection, is upregulated in the livers of malaria-infected mice. Chapter 3 shows that both activin B and the closely related protein activin A upregulate hepcidin in vitro and in vivo. Chapter 3 also explores the effects of the activin-binding protein follistatin in both systems and in the same malaria-infected mouse model as presented in Chapter 2. The work presented in Chapter 4 extends these studies to human infections by demonstrating that activin A protein co-increases with hepcidin in human serum during malaria infection. Taken together, these findings are consistent with a novel role for activin proteins in controlling hepcidin upregulation in the context of malaria infection. This work may form a basis for the development of novel therapeutics that speed recovery from malarial anemia by inhibiting activins’ actions. Chapter 5 examines the role of infected red blood cell-derived microparticles in the initial recognition of a P. falciparum malaria infection, and subsequent hepcidin upregulation. Microparticles stimulate production of cytokines from peripheral blood mononuclear cells (PBMC), which also upregulate activin A message in response to both microparticles and whole infected red blood cells. These data are consistent with a model in which malaria-derived stimuli such as microparticles trigger the systemic release of activin proteins, which then act on the liver to upregulate hepcidin. Evidence has shown that cytokine levels at birth are related to malaria risk. In Chapter 6, hepcidin is measured in cord blood samples from participants in a large-scale clinical study in a malaria-endemic area, and shown to be elevated in cord blood from neonates with a clinical history of placental malaria. Cord blood hepcidin is also compared to birth levels of iron markers and other cytokines, and future clinical outcomes. Finally, the contributions of DNA methylation levels to cord hepcidin and cytokine levels are assessed by comparison of CpG methylation, at sites in genes encoding hepcidin and cytokines, to the serum concentrations of the genes’ protein products. Several intriguing associations are noted which indicate a possible novel role for DNA methylation in the determination of birth cytokine and hepcidin levels. Chapter 7 synthesizes the data presented in this thesis, interprets the possible significance of the major findings, and offers suggestions for future work.

Dried blood spots is an alternative method to collect blood samples from research subjects. However, little is known about how hemoglobin and hematocrit affect bead-based multiplex assay performance. The purpose of this study was to determine how bead-based multiplex assays perform when analyzing dried blood spot samples. A series of four experiments outline the study each with a specific purpose. A total of 167 subject samples were collected and 92 different biomarkers were measured. Median fluorescence intensity results show a positive correlation between filtered and non-filtered samples. Utilizing a smaller quantity of sample results in a positive correlation to a larger sample. Removal of hemoglobin from the dried blood spot sample does not increase detection or concentration of biomarkers. Of the 92 different biomarkers measured 56 were detectable in 100-75% of the attempted samples. We conclude that blood biomarkers can be detected using bead-based multiplex assays. In addition, it is possible to utilize a smaller quantity of sample while avoiding the use of the entire sample, and maintaining a correlation to the total sample. While our method of hemoglobin was efficient it also removed the biomarkers we wished to analyze. Thus, an alternative method is necessary to determine if removing hemoglobin increases concentration of biomarkers. More research is necessary to determine if the biomarkers measured in this study can be measured over time or within an experimental model.

Cerebrospinal fluid is examined using a variety of methods, which also include determining the number and type of each cell. Now the method of the first choice to determine the cellular elements is a microscopic method. This determination, however, can also be done using the analyzer method, which is not yet so widespread. The aim of my thesis was to compare these two methods and determine whether the examination of cerebrospinal fluid on the analyzer Sysmex XE-5000 in the "Body Fluid" mode can replace commonly used microscopic methods. To this purpose, we gathered the laboratory data measured by using both these methods. These data was compared, evaluated and statistically processed. The resulting data suggest that the values measured on the analyzer Sysmex XE-5000 are more accurate than from microscopic determination, especially at highly cell samples of cerebrospinal fluid. To this end, we came evaluation of Bland-Altman graphs and comparison graphs with marked of limits of physiological oligocytosis. For checking of the accuracy of measurements, we verified the repeatability of the analyzer for the values of leukocytes and erythrocytes, coefficients of variation corresponding to the values specified by the manufacturer's documentation. We also investigated the stability of samples of...

Orientador: Elizabeth Moreira dos Santos Schmidt / Resumo: A ovinocultura brasileira apresentou crescimento nos últimos anos e é responsável pela produção de carne e leite, assim como lã e couro, e as infecções por nematódeos gastrointestinais são o maior problema sanitário. O objetivo do estudo foi avaliar a infecção natural por nematódeos gastrointestinais de borregas inseridas em um sistema de produção agropecuária. Borregas da raça Corriedale foram divididas em três grupos de dez animais cada, definidos de acordo com o OPG e hematócrito obtidos após a primeira coleta. O grupo 1 (G1) foi composto por animais com OPG (ovos por grama de fezes) maior do que 5.000 e hematócrito menor ou igual a 24%; o grupo 2 (G2) com OPG maior do que 5.000 e hematócrito maior do que 24% e o grupo 3 (G3) com OPG menor do que 5.000 e hematócrito maior do que 24%. Amostras de fezes (testados para OPG e coprocultura) e sangue (hemograma e a determinação de variáveis bioquímicas séricas) foram coletadas em quatro ao longo do período experimental de 70 dias. Ao final do estudo duas borregas foram abatidas para coleta e identificação de parasitas adultos. Todas as variáveis foram avaliadas pelo teste de normalidade e variância homogênea, posteriormente testada pelo ANOVA, e comparação de médias pelo teste Tukey. Os gêneros Haemonchus, Trichostrongylus e Cooperia foram recuperados em coprocultura. As contagens de OPG apresentaram diferenças significativas (P<0,05) nos três grupos, nos momentos 1 e 2. Os valores do hematócrito foram significativamente menores... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Brazilian sheep population has grown in recent years, and It's responsible for meat and milk production, as well as wool and leather, and gastrointestinal nematode infections are the major sanitary problem. Our aim was to evaluate natural infection by gastrointestinal nematodes of lambs inserted in a integrated crop-livestock system. Corriedale’s lambs were divided into three groups of ten animals each, defined according to the EPG (eggs per gram of faeces) and hematocrit obtained after the first collection. Group 1 (G1) was composed by animals with EPG greater than 5,000 and hematocrit less than or equal to 24%; group 2 (G2) with EPG greater than 5,000 and hematocrit greater than 24% and group 3 (G3) with EPG lower than 5,000 and hematocrit greater than 24%. Stool samples (tested for EPG and stool test) and blood (blood count and determination of serum biochemical variables) were collected over the 70 day trial period.. At the end of the study, two ewes were slaughtered for collect and identification of adult parasites. All variables were evaluated by the normality test and homogeneous variance, later tested by ANOVA, and a comparison of means by the Tukey test. Haemonchus, Trichostrongylus, and Cooperia genera were recovered in a stool test. EPG counts presented significant differences (P <0.05) in the three groups at moments 1 and 2. The hematocrit values were significantly lower (P <0.05) for G1 in M1 and G2 in M2. The variables CHCM, total leukocytes, lipase, cholesterol... (Complete abstract click electronic access below) / Mestre

OBJETIVO: Investigar o efeito da expressão de RNAm dos SMADs e de microRNAs (miRNAs) que possuem o TGFB1 como alvo na expressão gênica (RNAm e proteína) de TGF-&#946;1 e seu papel na fisiopatologia da angiogênese em pacientes com mielofibrose (MF) e trombocitemia essencial (TE). MÉTODOS: Foram incluídos 21 pacientes com MF primária (MFP), 21 com MF pós-TE (MFPTE) e 24 com TE, além de 98 indivíduos controles pareados de acordo com gênero e idade com os pacientes. As análises realizadas no sangue periférico foram: quantificação das concentrações plasmáticas e de RNAm de TGFB1, VEGFA e FGF2; quantificação de RNAm de SMADs 1 a 7 e de miRNAs 193a-5p, 369-5p, 542-5p, 590-3p, e 590- 5p; e detecção das mutações JAK2V617F (com quantificação alélica), MPLW515K/L e CALR. Em 26 biópsias de medula óssea dos pacientes, foram determinados o grau de microvasculatura (angiogênese estimada - CD34), a imunoexpressão de TGF-b1 ativo, TGF-&#946;1 latente e c-MPL. RESULTADOS: As concentrações de TGF- &#946;1 plasmático foram semelhantes entre os pacientes e controles, enquanto o VEGFA plasmático foi maior em todos os grupos de pacientes comparados aos seus controles. O FGF2 plasmático também foi maior em todos os grupos de pacientes, e a expressão de seu RNAm foi maior nos pacientes com TE do que em seus controles. As expressões de SMADs e de miRNAs foram semelhantes entre pacientes e controles. TGF-&#946;1 e FGF2 plasmáticos apresentaram correlações positivas nos pacientes com MFP, e correlações negativas nos seus controles, assim como nos controles de MFPTE. Em todos os grupos estudados foi observada correlação positiva entre TGF-&#946;1 e VEGFA plasmáticos. Além disso, foram demonstrados diferentes perfis de correlações entre a expressão gênica de TGF-&#946;1 e os diversos SMADs e miRNAs em cada grupo de pacientes e controles. Os pacientes com MFP com maior angiogênese (de acordo com a mediana da concentração plasmática de VEGFA e FGF2) apresentaram maiores concentrações plasmáticas de TGF-&#946;1 do que aqueles com menor angiogênese. A angiogênese medular estimada (CD34) não foi diferente entre os três grupos de pacientes estudados. Além disso, não foram encontradas correlações entre a imunoexpressão de CD34 e as expressões de RNAm de TGFB1, VEGFA e FGF2 medulares nem em leucócitos de sangue periférico, ou a concentrações plasmáticas de TGF-&#946;1, VEGFA e FGF2. As imunoexpressões de TGF-b1 ativo, TGF-&#946;1 latente e c-MPL foram semelhantes entre os três grupos de pacientes. As frequências das mutações avaliadas foram similares às descritas na literatura. Os pacientes com MFPTE portadores de mutação CALR apresentaram menores concentrações plasmáticas de VEGFA e FGF2 do que os JAK2V617F positivos, enquanto os pacientes com TE portadores de mutação CALR exibiram menores concentrações plasmáticas de TGF-&#946;1 do que os portadores de JAK2V617F. CONCLUSÕES: O presente trabalho permitiu confirmar a correlação positiva entre o TGF-&#946;1 com outros dois marcadores de angiogênese (VEGFA e FGF2). As expressões de SMADs e de miRNAs estudados foram semelhantes entre pacientes e controles, visto não haver diferenças na expressão gênica de TGF-&#946;1. Entretanto, disparidades encontradas nas correlações entre a expressão gênica de TGF-&#946;1 e diferentes SMADs e miRNAs nos pacientes e controles poderiam indicar que a regulação da expressão gênica de TGF-&#946;1 nas doenças estudadas seja distinta da apresentada nos indivíduos sem essas doenças. / AIM: To investigate the effects of the expression of SMADs mRNA and microRNAs (miRNAs) that target TGFB1 in TGF-&#946;1 gene expression (mRNA and protein) and its role in the angiogenesis pathophysiology in myelofibrosis (MF) and essential thrombocythemia (ET) patients. METHODS: Twenty-one primary MF (PMF), twenty-one MF post-ET (MPET) and twenty-four ET patients were included, besides 98 controls matched for gender and age with patients. In peripheral blood were assessed: TGF-&#946;1, VEGFA and FGF2 plasmatic levels and mRNA quantification; SMADs 1 to 7 mRNA quantification and miRNAs 193a-5p, 369-5p, 542-5p, 590-3p, and 590-5p quantification; and detection of JAK2V617F (and allele burden), MPLW515K/L and CALR mutations. Estimated angiogenesis (microvessel grade - CD34), active TGF-b1, latent TGF-&#946; and c-MPL immunoexpression were determined in 26 bone marrow biopsies. RESULTS: Plasmatic TGF-&#946;1 levels were similar in patients and controls, while all the patients groups had higher plasmatic VEGFA than controls. Plasmatic FGF2 was higher in all the patients groups, and its mRNA expression was higher in ET patients than in controls. No differences in SMADs and miRNAs expression were found between patients and controls. There was a positive correlation between plasmatic TGF-&#946;1 and FGF2 in PMF, and a negative correlation between these variables in their controls, as well as in MPET controls. In all studied groups, there was a positive correlation between plasmatic TGF-&#946;1 and VEGF. In addition, different profiles of correlations were demonstrated between TGF-&#946;1 gene expression and the several SMADs and miRNAs studied in each group of patients and controls. PMF patients with higher angiogenesis (according to the median of VEGFA and FGF2 plasma levels) had higher plasmatic TGF-&#946;1 levels than those with lower angiogenesis. Estimated angiogenesis (CD34) in bone marrow biopsies were not different among PMF, MPET and ET patients. Moreover, there were no correlation between CD34 immunoexpression and TGFB1, VEGFA and FGF2 mRNA bone marrow or peripheral blood expression or plasmatic levels, as well as latent TGF-&#946;1, active TGF-b1, and c-MPL immunoexpression were similar in patients studied groups. The frequencies of evaluated mutations were similar to previously reported. MPET patients harboring CALR mutations had lower plasmatic VEGFA and FGF2 than JAK2V617F mutated, while ET patients carrying CALR mutations had lower plasmatic TGF-&#946;1 than JAK2V617F mutated. CONCLUSIONS: This study confirmed the positive correlation among TGF-&#946;1 and two other markers of angiogenesis (VEGFA and FGF2). SMADs and miRNAs expressions were similar between patients and controls, since there were no differences in TGF-&#946;1 gene expression between patients and controls. However, disparities found in the correlations between TGF-&#946;1 gene expression and different SMADs and miRNAs in patients and controls may indicate that TGF-&#946;1 gene expression regulation in studied diseases is distinct from those presented by individuals without these diseases.

Phrynops geoffroanus é uma espécie de quelônio com ampla distribuição na América do Sul, ocupando diversos tipos de habitat, inclusive rios urbanos poluídos. Com o objetivo de se conhecer aspectos ecológicos da espécie em ambientes antropizados, durante os anos de 2006 e 2007 foi realizado um estudo na bacia do rio Piracicaba, tendo como sítios amostrais o rio Piracicaba e seu afluente o ribeirão Piracicamirim. Um dos temas deste estudo foi à avaliação de aspectos sanitários da espécie. Foi investigado o perfil hematológico dos cágados no rio Piracicaba e ribeirão Piracicamirim, e avaliado um índice de estresse (razão heterófilo/linfócito), que além dos dois corpos d\'água foi adicionado um grupo de animais provenientes de cativeiro, da Fundação Parque Zoológico de São Paulo. A diferença entre um rio de maior volume, Piracicaba, e seu afluente Piracicamirim, ambos poluídos e com diferentes históricos de ocupação do solo, pouco influenciaram nas variações dos dados hematológicos dos cágados e os animais não apresentaram nenhum sinal de enfermidade através dos resultados obtidos no hemograma. Estas análises foram o tema do primeiro capítulo desta dissertação. Outro aspecto sanitário avaliado foi a microbiota bacteriana oral dos cágados em ambos os corpos d\'água, abordado no segundo capítulo. As amostras foram incubadas a 25 e 37ºC. Avaliou-se a patogenicidade das bactérias tanto para os cágados quanto para seres humanos. Houve crescimento de bactérias patogênicas para o homem e para os animais e a bactéria mais prevalente a 37ºC foi a Escherichia coli. Apesar disso, os cágados não apresentaram sinal aparente de infecção. Estudos de longo prazo são recomendados para animais residentes em ambientes antropizados, monitorando-se o estado sanitário e aspectos ecológicos e demográficos. / Phrynops geoffroanus is a freshwater turtle species, with a wide distribution in South America, living in many types of habitats, including polluted urban rivers. During 2006 and 2007, aiming to study ecological aspects of the species in anthropogenic environments, it was conducted an investigation at Piracicaba River Basin, in two sample sites, the Piracicaba river and its tributary, Piracicamirim stream. One of the topics studied was a health assessment of the turtles. It was examined the blood profile in Piracicaba and Piracicamirim turtles and evaluated a measure of stress (heterophil/lymphocyte ratio), that had another study group besides the animals from the rivers, turtles in captivity from Sao Paulo Zoo Park Foundation. The difference between a large river, Piracicaba and its tributary Piracicamirim, both polluted and with different soil use and occupation had a little influence on turtles\' hematological variation, and the animals did not show any sign of disease through the blood profile examination. These evaluations were the subject of the thesis\' first chapter. Another part of the health assessment study, subject of the second chapter, was the oral microbiota investigation, performed in Piracicaba and Piracicamirim turtles. The samples were incubated at 25 and 37º Celsius, aiming to check the bacterium\'s pathogenicity for the animals and for humans. The results showed growth of pathogenic bacterium for both and the most prevalent bacteria at 37ºC was Escherichia coli. Although, the turtles did not show any sign of infection. Long-term studies are suggested for turtles living in anthropogenic environments to monitor their health status, demography and ecological issues.

O presente trabalho teve como objetivo avaliar a influência da cesariana eletiva no eritrograma de bezerros neonatos da raça Nelore durante os primeiros trinta dias de vida. Os neonatos foram alocados em 3 grupos 8 bezerros obtidos por parto espontâneo (PE), 8 bezerros nascidos por cesariana, sem indução do parto (CSI) e 8 bezerros nascidos por cesariana, com indução do parto (CCI) pelo uso de 25 mg de Dexametasona, 24 horas antes do procedimento cirúrgico, pela via intramuscular. Os nascimentos ocorreram entre 282 e 292 dias de gestação. As amostras de sangue para a determinação do quadro eritrocitário foram colhidas por punção da veia jugular externa, utilizando-se Sistema Vacutainer&reg;. A determinação do eritrograma foi realizada no contador automatizado BC-2800 Vet Mindray&reg;. Os resultados obtidos evidenciam que a cesariana eletiva não influenciou o número de hemácias, das taxas de hemoglobina e nos valores do volume globular a crase sanguínea de bezerros na primeira semana de vida. Na primeira semana de vida observou-se, independentemente do grupo experimental uma diminuição no número de hemácias, das taxas de hemoglobina e nos valores do volume globular. Até os 5 dias de vida, os valores de Volume Corpuscular Médio (VCM) e de Hemoglobina Corpuscular Média (HCM) dos bezerros nascidos de partos espontâneos por via vaginal são estatisticamente menores do que os obtidos em bezerros nascidos de cesariana eletiva. A partir de 10 dias de vida observou-se, nos animais obtidos de parto espontâneo, uma rápida recuperação nos valores de hemácias, hemoglobina e volume globular, enquanto nos animais nascidos por cesariana (induzida e não induzida) não ocorria essa recuperação nas taxas de hemoglobina e nos valores do volume globular. Nos bezerros obtidos por cesariana observou-se que no período entre 10 e 30 dias de vida o Volume Corpuscular Médio (VCM) e a Hemoglobina Corpuscular Média (HCM) diminuíram, passando a apresentarem microcitose e hipocromia quando comparados com bezerros obtidos por parto espontâneo. Entre todos os grupos, os animais nascidos por cesariana sem indução apresentaram os valores com menor magnitude do início ao fim do período experimental. Os animais nascidos a termo demonstram uma capacidade de recuperação fisiológica mais apurada que dos animais nascidos por cesariana. / This study aimed to evaluate the influence of elective cesarean section in erythrogram of newborn calves of the Nellore breed for the first thirty days of life. Neonates were divided into 3 groups 8 calves obtained by spontaneous birth (PE), 8 calves born by caesarean section without labor induction (CSI) and 8 calves born by cesarean section with induction of labor (CCI) by the use of 25 mg of dexamethasone 24 hours before the surgical procedure, by the intra-muscular). Births occurred between 282 and 292 days of gestation. Blood samples for determining the erythrocyte frame were collected by puncture of the external jugular vein using Vacutainer &trade; System. The determination of erythrocyte was performed in the automated counter BC-2800 Vet Mindray&reg;. The results show that elective caesarean section did no affect the number of red blood cells, hemoglobin rates and values of the globular volume in the first week of life of calves. At this first week of life was observed, regardless of the experimental group a decrease in the number of red blood cells, hemoglobin rates and values of the globular volume. Until 5 days of age, the mean corpuscular volume values (MCV) and Mean Corpuscular Hemoglobin (MCH) of calves born from spontaneous vaginal deliveries are statistically lower than those obtained in calves born from elective cesarean section. From 10 days of life was observed in animals obtained by from spontaneous delivery, a quick recovery in the values of erythrocytes, hemoglobin and packed cell volume, whereas in animals born via caesarean section (induced and uninduced) did not occur such recovery in the rates of hemoglobin and packed cell volume values. In calves obtained by Caesarean section it was observed that in the period between 10 and 30 days of life Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin (MCH) decreased, submitting a microcytic hypochromic when compared with calves obtained by spontaneous delivery. Among all groups, animals born by caesarean section without induction showed the values with lower magnitude from beginning to end of the trial period. Animals born at term show a more accurate physiological recovery capability in comparingson with animals born by Caesarean section.