Orientadores: Marlene Tiduko Ueta, Eleonore Zulnara Freire Setz / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-05T12:53:18Z (GMT). No. of bitstreams: 1 Rondon_MichelleVivianeSadosSantos_M.pdf: 2874588 bytes, checksum: 28e75d1c2516ff11dfb2158b272c510f (MD5) Previous issue date: 2005 / Resumo: Durante o período de agosto de 2003 a julho de 2004 foram realizadas coletas mensais de fezes de bugios-ruivos, Alouatta guariba (Primates, Atelidae), em um fragmento de Floresta Atlântica Semidecídua, a Mata Ribeirão Cachoeira, situada no Distrito de Souzas ¿ Campinas/SP, com 234 hectares. As fezes foram coletadas ao longo de duas trilhas longitudinais dentro do fragmento florestal, um pela calha do riacho e outro por uma trilha paralela com 4.000m de comprimento, com o objetivo de realizar um estudo de helmintos intestinais nestes animais. Foram realizadas 13 coletas com um total de 112 amostras, entre fezes frescas (N=102) e secas (N=10), que foram processadas qualitativamente pelos métodos de sedimentação Hoffman e Rugai e pelos métodos de flutuação Faust e Willis. Das 112 amostras, 61 foram positivas para ovos, larvas ou adultos de nematódeos, representando 54,5%. Os tipos morfológicos de nematódeos encontrados, foram identificados neste trabalho, pela numeração seqüencial, conforme foram sendo observados. Foram encontrados sete tipos morfológicos de ovos, nove tipos morfológicos de larvas e sete de nematódeos adultos. Os ovos tipo 2 de Trypanoxyuris (Trypanoxyuris) minutus (Oxyuridae), foram encontrados em fezes nos dois percursos em nove coletas. Ovos tipo 1 foram pouco freqüentes (N=5) e observados somente em um dos meses de coleta (agosto/2003); são ovos com características morfológicas compatíveis a Strongyloides sp., já relatado para primatas do gênero Alouatta mas, devido ao pequeno número, não houve confirmação. O nematódeo adulto tipo 1 (Oxyuridae, Cosmocercinae) foi o mais comum, ocorrendo em sete coletas no riacho e duas coletas na trilha, tanto em fezes frescas, como secas. O adulto tipo 7, de Trypanoxyuris (Trypanoxyuris) minutus foi observado somente em uma coleta (junho/2004), na trilha paralela. Os outros cinco tipos morfológicos de nematódeos adultos, foram encontrados entre os meses de abril e junho/2004 na trilha, exceto o adulto tipo 6 que foi encontrado no riacho. As amostras fecais mostraram-se positivas para alguma forma de nematódeo em dez coletas do período de estudo / Abstract: From August 2003 to July 2004 I collected fecal samples from the Brown-howler-monkey Alouatta guariba (Primates, Atelidae) monthly in a 234 ha Semideciduous Atlantic Forest fragment (Mata Ribeirao Cachoeira - Campinas/SP) in order to identify intestinal helminths. Feces were collected along two transects inside the forest fragment, one following the stream and the other on a parallel trail, each 4 km long, with the purpose of examining howlers intestinal helminths. On 13 sampling trips 112 samples (102 fresh and 10 dry scats) were collected. All the scats were processed qualitatively by sedimentation (methods of Hoffman and Rugai), and by flotation (Faust and Willis). Of the 112 samples, 61 (54.5%) were positive for eggs, larvae or adults from Nematoda. Morphotypes were enumerated chronologically as they appeared in the samples. There were seven egg morphotypes, nine larvae morphotypes and seven for adults, all Nematoda. Eggs of Trypanoxyuris (Trypanoxyuris) minutus (Oxyuridae) were found in feces along both transects on nine dates. Egg type 1 was uncommon (N=5) and only observed in August 2003. They shared morphological characteristics with Strongyloides sp., already reported for the genus Alouatta. Due to the small sample, confirmation was not possible. A species of Oxyuridae (Cosmocercinae) was the most common adult nematode, occurring on seven dates in the stream transect and twice on the trail. It was found both in fresh and dry scats. Adult Trypanoxyuris (Trypanoxyuris) minutus were observed once (June 2004), on the trail. The additional five adult nematoda morphotypes were found mostly along the trail between April and June 2004, except for the adult type 6 that was found along the stream. The fecal samples were positive for one or more Nematoda on ten sample dates year around / Mestrado / Doutor em Parasitologia

Retrovírus endógenos (ERVs) são vírus que infectaram células germinativas e proliferaram no genoma do hospedeiro. A família K está integrada apenas no genoma de primatas do Velho Mundo. Os ERVs promovem alterações estruturais nos genomas hospedeiros, sendo fundamentais para a sua evolução. Esse trabalho teve como objetivo realizar uma investigação da distribuição e dinâmica evolutiva de ERV-K nos diferentes hospedeiros. Foram identificados 58 ERV-K em humanos, 38 em chimpanzés, 35 em orangotangos e 19 em macaco rhesus. Análises filogenéticas evidenciaram dois grupos principais, Grupo O/N, que compreende os provirus mais antigos e os mais recentes, e Grupo I, com provirus com tempo de integração intermediário. A dinâmica de espalhamento de ERV-K diferiu entre os hospedeiros. A fixação e eliminação dos ERV-K é resultado de fatores demográficos e populacionais, como gargalos de garrafa e expansões sofridas ao longo da evolução. Análises de quais provírus são ativos em pacientes com HIV e com cancer demonstrou que distintos ERVs são transativados, sugerindo alguma consequencia biológica para o hospedeiro. Além disso, a atividade dos ERVs não depende exclusivamente do tempo de integração, mas sim da integridade de regiões específicas contidas na LTR. / Endogenous retroviruses (ERVs) are remains of ancient viral infection in the germ line cells and subsequent vertical transmission. The K family are integrated only in humans and the Old World monkeys. ERVs play a fundamental role on genome evolution and foster variability. The aim of this work was to investigate their distribution and evolutionary dynamics in primate hosts. We found 55 ERV-K genomes in the human genome, 38 in chimpanzee, 35 in orangutan and 19 in Rhesus monkey. Two main groups were recovered by phylogenetic inference, Group O/N, comprising the newest and the oldest proviruses and, Group I, enclosing those with intermediate integration time. Although the primary integration took place in the ancestral lineage of all primates investigated, their evolutionary dynamic was different among them. I propose that ERV-K dynamics depends on the host demography experienced throughout their evolution. This work also investigated the putative source of proviral transcripts detected in HIV carries and cancer patients. The differential expression found under these conditions suggested a biological role of the ERV-K overexpression. Finally, the results showed that the ERV-K overexpression depends on the integrity of specific promoters in their LTR.

Originalmente da Bahia, a espécie Leontopithecus chrysomelas foi introduzida no estado do Rio de Janeiro provavelmente por meio do tráfico e atualmente é encontrado em uma das áreas remanescentes de ocupação natural do mico-leão-dourado (Leontopithecus rosalia) podendo comprometer a sobrevivência do último. Visando mitigar esse problema foi criado um plano de ação, onde se recomenda a captura dos animais introduzidos. Procedimentos de captura e contenção química produzem estresse e afetam diretamente a homeostase, podendo alterar diretamente a saúde e o bem estar dos animais somando-se o fato de os protocolos de capturas atuais gerarem um período de recuperação prolongado. O objetivo deste trabalho consistiu em determinar e comparar os efeitos cardiorrespiratórios da dexmedetomidina, quando associada à cetamina S(+) ou à cetamina racêmica, e do midazolam associado à cetamina S(+), por via intramuscular, na contenção química de mico-leão-da-cara-dourada. Foram utilizados 45 animais da espécie Leontopithecus chrysomelas, adultos, machos, pesando em média 532 gramas, oriundos do Centro de Primatologia do Rio de Janeiro, em parceria com a ONG PRI-MATAS. Os animais foram distribuídos em três grupos onde receberam aleatoriamente as associações: CSM - cetamina S(+) (15 mg/kg) e midazolam (0,5 mg/kg); CSD - cetamina S(+) (15 mg/kg) e dexmedetomidina (10 µg/kg); e CD - cetamina racêmica (15 mg/kg) e dexmedetomidina (10 µg/kg). Foram avaliados os períodos de latência, hábil e de recuperação. A frequência e ritmo cardíacos, frequência respiratória, saturação da oxihemoglobina periférica, pressão arterial sistólica, temperatura retal, qualidade de indução e de recuperação, sedação, antinocicepção e grau de relaxamento muscular foram monitorados a cada cinco minutos durante 50 minutos. Os dados paramétricos foram avaliados pela análise de variância (ANOVA) seguida do teste de Tukey. Os dados não paramétricos foram avaliados pelo teste de Kruskal-Wallis seguido do teste de Dunn para identificar quais grupos apresentavam diferença estatística significativa. O grau de significância estabelecido para as análises foi de 5% (p<0,05). Os valores referentes aos parâmetros de frequência respiratória, saturação da oxihemoglobina periférica, temperatura, pressão arterial sistólica, início da perda de tônus muscular, latência e os períodos de recuperação parcial e total não apresentaram diferença entre os grupos. Já os valores de frequência cardíaca, relaxamento muscular, antinocicepção, grau de sedação e consumo de lidocaína apresentaram diferença significativa entre os mesmos momentos dos grupos CSM e CSD, e CSM e CD. Conclui-se que as associações de dexmedetomidina com cetamina racêmica e cetamina S(+) apresentaram os melhores índices de relaxamento muscular, sedação e antinocicepção e mostraram-se seguras para a realização de cirurgia de vasectomia em micos-leão-da-cara-dourada. Apesar dos grupos CSD e CD apresentarem bradicardia mais acentuada, os valores mantiveram-se dentro do limite esperado. / The Golden-headed lion tamarin, originally endemic to the southern Bahia region, was introduced in the state of Rio de Janeiro probably by illegal wildlife trade activity. The species is currently found in one of the remaining areas of natural occupation of the golden lion tamarin (Leontopithecus rosalia), which may compromise the survival of the latter. In order to mitigate this problem, an action plan was elaborated and the capture of golden-headed lion tamarins recommended. Both capture and chemical restraint procedures can cause stress and compromise homeostasis with a direct effect on the health and well-being of the animals. Besides that, current chemical immobilization protocols usually result in prolonged recovery times. This study aims to determine and compare the cardiovascular and respiratory effects of three different protocols for chemical immobilization of golden-headed lion tamarins. Thirty-five adult male specimens of Leontopithecus chrysomelas were studied. Animals were randomly separated into three groups: ketamine S(+) and midazolam (15 mg kg-1 and 0.5 mg kg--1-) (KSM group), ketamine S(+) and dexmedetomidine (15 mg kg--1- and 10 µg kg-1) (KSD group) and racemic ketamine and dexmedetomidine (15 mg kg-1 and 10 µg kg-1 ) (KD group). Periods of latency, immobilization and recovery were evaluated. Heart rate and rhythm, respiratory rate, peripheral oxyhemoglobin saturation, systolic blood pressure, rectal temperature, induction and recovery quality, sedation, antinociception, and degree of muscle relaxation were monitored every five minutes for 50 minutes. Parametric data were analyzed by using repeated measures analysis of variance (ANOVA) followed by a Tukey\'s test. Non-parametric data were analyzed by the Kruskal-Wallis test followed by Dunn\'s test. Values of P< 0.05 were considered statistically significant. No significant differences were found in respiratory rate, peripheral oxygen saturation, temperature, systolic blood pressure, onset of muscle tone loss, latency and partial and total recovery periods. Heart rate, sedation and muscle relaxation degrees, antinociception, and lidocaine consumption presented significant difference at the same moments between KSM and KSD and between KSM and KD. Marked bradycardia was presented on KSD and KD groups, with values remaining within the normal range. This study demonstrated that combinations of dexmedetomidine with racemic ketamine and S(+) ketamine presents the best outcomes for muscle relaxation, sedation and antinociception and were safe for vasectomy surgery in golden-headed lion tamarins.

Magister Scientiae - MSc / Male infertility in humans has increased in the last few decades and could be as high as 40%, while up to 50% of these men have ''unexplained'' (idiopathic) infertility. Although newly developed molecular techniques have great value in detecting subtle causes of male infertility, more detailed sperm functional tests are required to identify compromised fertility, especially in a clinical set-up. Since ethical constraints often preclude the pursuit of many basic research questions in humans, non-human primates (NHPs) have been identified as key models in human-related studies. NHPs are often used in studies on male fertility/infertility, IVF or assisted reproductive technology (ART) procedures, male contraception and reproductive toxicology. However, comparing results of NHP and human studies require that techniques used for assessment must be objective, standardized and sensitive to recognize compromised sperm function. The aim of this study was to evaluate standard sperm functional tests and develop new functional tests using NHP sperm, specifically from vervet monkeys (Chlorocebus aethiops), chacma baboons (Papio ursinus) and rhesus monkeys (Macaca mulatta), for application in human and NHP studies and to ultimately develop a basic primate model. The sperm functions investigated included sperm motility, longevity, vitality, DNA integrity, acrosome reaction, and hyperactivation. The sperm functional tests evaluated were: CASA motility analysis; Sperm Longevity test; Eosin-Nigrosin and Hoechst and Propidium Iodide staining, as well as the use of WST-1 cytotoxicity assay for vitality; the TUNEL assay for DNA integrity; Acrosome Intactness Test; and induction of hyperactivation via stimulants. The validity of each test was investigated by inhibiting sperm function through the use of copper sulphate and cadmium chloride. All functional tests were successfully performed across all three species, except the TUNEL assay for DNA integrity, and was further used for validation testing. Validation testing proved that all sperm functional parameters were significantly affected by the highest concentrations of the chemicals (250 µg/ml CuSO4 and 500 µg/ml CdCl2) and if not significant, trends of reduction were seen. The tests employed were therefore sensitive to the inhibitory effect of the metals. By evaluating these established sperm functional tests we found that primates would serve as good models for research study. Furthermore, we optimized and modified techniques for sperm and functional analysis in these three primate species and this study will standardize protocols for use in future studies on male infertility. Additionally, comparing human and NHP sperm function can possibly reveal or explain the high infertility rates in humans.

Species vary in the ease with which they can solve apparently similar problems. For instance, problems will be interpreted differently by different species due to differences such as how they process the world or their ecology. The latter is the focus of the ecological approach to cognition, which posits that ecology influences decision-making such that each species performs better on tasks that are naturally relevant to them. In a previous work, my colleagues and I compared the performance of cleaner fish and nonhuman primates, which differ substantially in brain size and ecology, on a dichotomous choice task derived from the cleaner fish ecology. In the task, subjects chose between two different plates, each containing a same food; if they chose the food from plate A, they could then choose the food from plate B as well, whereas if they chose B first, A was no longer available. Fish were better than primates at solving this task, emphasizing the role of ecology in shaping one species’ decision-making. For my dissertation project, I explored possible explanations for the primates’ poor performance in the task. In a first series of studies, I investigated the possibility that species differed in the task because of differences in the capacity to recognize the relevant cues (i.e. the plate design), to solve it. I thus repeated the task with fish and nonhuman primates, using variations designed to be more salient to primates. In a first experiment, the foods were different colors, whereas in a second experiment, they were hidden to avoid the prepotent response. In a second series of studies, I tested monkeys in a computerized paradigm that differed from the plate task by removing interaction with the human experimenter, which may be distracting, and providing a more standardized testing environment. Finally, in a last study, I investigated one possible cognitive limitation to the primates in the plate task, the failure to use backwards induction to solve it. These studies allowed me to investigate the role of ecology in species’ decision-making, a perspective that is often neglected in studies of cognition.

Siglecs and CEA are two families of cell surface proteins belonging to the immunoglobulin superfamily. They are thought to be involved in cell-cell interactions and have various other biological functions. We used the GENECONV program that applies statistical tests to detect gene conversion events in each family of five primate species. For the Siglec family, we found that gene conversions are frequent between CD33rSiglec genes, but are absent between their conserved Siglec genes. For the CEA family, half of gene conversion events detected are located in coding regions. A significant positive correlation was found between the length of the conversions and the similarity of the converted regions only in the Siglec gene family. Moreover, we found an increase in GC-content and similarity in converted regions compared to non-converted regions of the two families. Furthermore, in the two families, gene conversions occur more frequently in the extracellular domains of proteins, and rarely in their transmembrane and cytoplasmic regions. Finally, these two families appear to be evolving neutrally or under negative selection.

A endocrinologia reprodutiva do muriqui do sul B. arachnoides foi avaliada em quatro fêmeas adultas pela dosagem de metabólitos fecais de estrógenos e em quatro machos adultos e um macho subadulto pela dosagem de metabólitos de testosterona e glicocorticóides durante um período de onze meses em duas diferentes condições restritivas, ilha de 600m2, Curitiba (PPC) e viveiro (15,40x5,85x4,70m) Rio de Janeiro (CPRJ). As fêmeas apresentaram grande variação individual nas concentrações de estrógenos e progestinas fecais ao longo do período de estudo, três não apresentaram atividade ovariana no período de outubro-dezembro e apenas uma apresentou atividade durante todos os períodos amostrados. Mesmo com níveis baixos de esteróides as fêmeas apresentaram comportamento perceptivo e cópulas. Os machos do PPC apresentaram níveis significativamente mais altos de glicocorticóides e níveis significativamente mais baixos de testosterona do que os machos do CPRJ (p<0,05) apresentaram. O macho subadulto apresentou níveis significativamente mais baixos para metabólitos de testosterona. Nos machos as cópulas ocorreram próximo às elevações das concentrações de testosterona. Em três situações grandes elevações dos níveis de glicocorticóides fecais puderam ser relacionadas a situações estressantes. Nosso estudo comprovou a eficácia do método empregado para monitoramento reprodutivo bem como para avaliar situações estressantes. / The reproductive endocrinology of the southern muriqui (Brachyteles. Arachnoids) was evaluated through the dosage of fecal metabolites steroids for 04 adult females and fecal metabolites of testosterone and glucocorticoids for 04 adult males and 01 subadult male. The study was conducted over an eleven month period at two environmental conditions: a) an island of 600 m2, with natural vegetation, at Curitiba Zoo (PPC) and b) a large cage of 15,40X5,85X4,70m at Rio de Janeiro Primatological Centre (CPRJ). It was observed that females had large strogen interindividual variation and fecal progestins: three females did not show ovarian activity over a partial period of the study, while ovarian activity was observed for all sample periods for one female. Despite the detection of low levels of steroids, proceptive behaviour and copulations were observed. The PPC males showed significant levels of glicorticoids and testosteorne when compared to the CPRJ males (p<0.05). The subadult male exhibited the lowest level of testosterone metabolites and those differences were significant when compared to the adult males (p<0.05). For all males, copulations occurred when testosterone levels were highest and peaks of glucorticoids were linked to stressfull situations. This study have shown that the method used was effective for reproductive monitoring as well as for evaluating stressfull situations.

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付記する学位プログラム名: 霊長類学・ワイルドライフサイエンス・リーディング大学院 / 京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第21615号 / 理博第4522号 / 新制||理||1649(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)准教授 Andrew MacIntosh, 教授 古市 剛史, 教授 髙井 正成 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DFAM

付記する学位プログラム名: 霊長類学・ワイルドライフサイエンス・リーディング大学院 / 京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第22635号 / 理博第4624号 / 新制||理||1664(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)准教授 足立 幾磨, 准教授 後藤 幸織, 教授 濱田 穣 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM