An intensive, replicated monthly sampling of snake communities inhabiting four habitat types was conducted at John F. Kennedy Space Center, Brevard County, Florida from November 2002 through October 2003. Thirteen species (580 individuals plus 74 recaptures) plus one hybrid were captured. The three most commonly captured species, Coluber constrictor, Thamnophis sirtalis, and Thamnophis sauritus, combined made up 85% of the sample. These three species were active during every month of the year, but showed modal activity patterns typical of Temperate Zone snakes. Monthly snake captures were correlated with monthly captures of potential prey species and with mean monthly temperature. Species richness in the four habitat types varied from nine to 12. Drift fences in ruderal habitats had the highest species richness, while fences in swales captured the greatest number of individuals. The most dissimilar habitat pair was scrub and swale, while the most similar pair was ruderal and hammock. Box traps proved more effective for targeting the largest snake species, while funnel and box traps were equally effective for targeting other species.

No description available.

Peninsula ribbon snakes, Thamnophis sauritus sackenii, were collected for 14 months. Of 90 individuals collected and examined, 42 were females, 29 were males, and 19 were neonates. Females had a larger body length than did males. The smallest, sexually mature female had a snout-vent length of 428 mm. Mating occurred from March through May. Parturition occurred from June through October. Vitellogenesis began in spring and ovulation occurred between April and May. Spermatogenic activity occurred from August through December and testes were regressed during other months.

Chrysopelea paradisi, also known as the flying snake, possesses one of the most unique forms of aerial locomotion found in nature, using its entire body as a dynamic lifting surface without the use of wings or membranes. Unlike other airborne creatures, this species lacks appendages to aid in controlling its flight trajectory and producing lift. The snake exhibits exception gliding and maneuvering capabilities compared with other species of gliders despite this lack of appendages. While gliding, C. paradisi morphs its body by expanding its ribs, essentially doubling its width and utilizing its entire length as a reconfigurable wing. Its cross-sectional shape transforms into a thick, airfoil shape with a concave ventral surface, outwards protruding lips at the leading and trailing edges, a somewhat triangular dorsal surface with a round apex, and fore-aft symmetry. This study investigated the aerodynamic performance of this unique shape by simulating a single, static segment of the snake's body over a wide range of Reynolds numbers (3,000 to 15,000) and angles of attack (-10 to 60o) to simulate the full range of the snake's flight kinematics. This is the first study on an anatomically accurate snake model, and few aerodynamic studies have been performed in this low Reynolds number regime. Load cell measurements and time-resolved digital particle image velocimetry (TRDPIV) were performed on a 2D anatomically accurate model to determine the lift and drag coefficients, wake dynamics, and vortex shedding characteristics. This geometry produced a maximum lift coefficient of 1.9 and maximum lift to drag ratio of 2.7, and maintained increases in lift up to 35o. Overall, this geometry demonstrated robust aerodynamic behavior by maintain significant lift production and near maximum lift to drag ratios over a wide range of test parameters. These aerodynamic characteristics may enable the flying snake to glide at steep angles and over a wide range of angles of attack, often encountered in gliding trajectories. This geometry also produced larger maximum lift coefficients than many other bluff bodies and airfoils in this low Reynolds number regime. This thesis is organized as follows. The first section contains a broad introduction on gliding flight and C. paradisi's unique mode of gliding. The following section is a manuscript that will be submitted to a journal and contains the experimental analysis on the snake's cross-sectional shape. Several appendices attached to the end of this thesis contain additional analysis and work performed throughout the duration of this project and unique Matlab algorithms developed during this research. / Master of Science

Future generations of unmanned spacecraft, aircraft, ground, and submersible vehicles will require precise relative navigation capabilities to accomplish missions such as formation operations and autonomous rendezvous and docking. The development of relative navigation sensing and control techniques is quite challenging, in part because of the difficulty of accurately simulating the physical relative navigation problems in which the control systems are designed to operate. A hardware testbed that can simulate the complex relative motion of many different relative navigation problems is being developed. This testbed simulates near-planar relative motion by using software to prescribe the motion of an unmanned ground vehicle and provides the attached sensor packages with realistic relative motion. This testbed is designed to operate over a wide variety of conditions in both indoor and outdoor environments, at short and long ranges, and its modular design allows it to easily test many different sensing and control technologies. / Master of Science

Os colubrídeos tem sido responsáveis por diversos acidentes graves, inclusive fatais, em seres humanos por todo o mundo. No Hospital Vital Brazil do Instituto Butantan, em São Paulo, 40% dos acidentes ofídicos registrados são causados por serpentes consideradas não-peçonhentas. Dentre estas, 97,3% pertencem à família Colubridae, onde 54,5% apresentam dentição áglifa e 42,8% dentição opistóglifa. Contudo, alguns acidentes envolvendo os colubrídeos opistóglifos podem desencadear manifestações clínicas semelhante ao acidente botrópico. Este fato tem causado muitas discussões, pois devido a um diagnóstico errôneo ou pela gravidade do acidente, muitas vezes é administrado o soro anti-botrópico, que em muitos casos pode ser ineficaz ou até prejudicial ao paciente. Vale ressaltar que raros são os estudos relacionados ao aparelho de \"veneno\" dos colubrídeos. Deste modo, tivemos como objetivo estudar as características anatômicas, morfológicas e ultra-estruturais da glândula de Duvernoy de seis espécies de colubrídeos opistóglifos. São eles, Philodryas olfersii e Philodryas patagoniensis da Tribo Philodryadini, Oxyrhopus guibei e Phimophis guerrini da Tribo Pseudoboini e Thamnodynastes strigatus e Tomodon dorsatus da Tribo Tachymenini. As glândulas de Duvernoy das espécies em questão foram analisadas macroscopicamente para descrição das características anatômicas. Foram então, fixadas em líquido de Bouin para posterior processamento histológico para análise morfológica e histoquímica. Para o estudo ultra-estrutural, as glândulas de Duvernoy foram fixadas em glutaraldeído 2,0% em tampão cacodilato 1,0M (pH 7,3) para posterior preparação de acordo com o protocolo pré-estabelecido. Os resultados obtidos mostraram que a referida glândula apresenta duas porções bem distintas, sendo a glândula supra-labial e a glândula de Duvernoy propriamente dita, as quais variam de forma e tamanho segundo a espécie. A glândula de Duvernoy é constituída basicamente por túbulos secretores e ductos excretores. As células dos túbulos secretores podem produzir substâncias de diferentes composições químicas, dependendo da espécie. Os resultados histoquímicos mostraram que a maioria das espécies analisadas apresenta uma glândula de natureza seromucosa, e apenas uma espécie teve a glândula de Duvernoy classificada como mucoserosa. Os grânulos de secreção presentes no citoplasma das células secretoras, observados nas eletromicrografias, corroboram os resultados histoquímicos, já que a eletrondensidade dos mesmos caracteriza a natureza das células. A função da glândula de Duvernoy está diretamente relacionada ao hábito alimentar destas serpentes e ao modo pelo qual subjugam as suas presas. As serpentes que apresentam a glândula de Duvernoy com características mucoserosas subjugam as suas presas por constricção. Já as espécies que possuem a glândula de Duvernoy com natureza seromucosa utilizam o processo de envenenamento das presas para se alimentarem. Deste modo, as serpentes garantem o sucesso na imobilização das presas, impedindo que estas utilizem táticas defensivas perigosas à serpente, independente do uso da constricção. / The colubrids have often been responsible for severe accidents, sometimes fatal for humans, all over the world. In the Hospital Vital Brazil of the Institute Butantan, in São Paulo, 40% of the snakebite registered is caused by serpents that are considered no poisonous. Among these, 97.3% belong to Colubridae family with 54.5% presenting aglyphous dentition and 42.8%, opisthoglyphous dentition. However, some accidents that involve the opisthoglyphous colubrids can break out clinic manifestation similar to the botropic accident. This fact has caused many debates, because due to an erroneous diagnosis or to the gravity of the accident, very often it is administered the anti-botropic serum, which in many cases may be ineffective or even harmful for the patient. It is important to mention that rare are the studies related to the venon-delivery-system of the colubrids. Thus, we propose to study the anatomical, morphological, histochemical and ultra structural characteristics of the Duvernoy´s gland of six species of opisthoglyphous colubrids. They are Philodryas olfersii and Philodryas patagoniensis of Tribe Philodryadini, Oxyrhopus guibei and Phimophis guerrini of Tribe Pseudoboini and Thamnodynastes strigatus and Tomodon dorsatus of Tribe Tachymenini. The Duvernoy´s glands of the species in question were analyzed macroscopically for description of the anatomical characteristics. They were then fixed in Bouin´s liquid for posterior histological procedures for morphologic and histochemical analysis. For the ultra structural study, the Duvernoy´s glands were fixed in glutaraldehyde 2.0% in buffer cacodylate 1.0 M (pH 7.3) for further preparation according to the established protocol. The results showed that the above-mentioned gland presents two well distinct portions, the salivary supralabial gland and the Duvernoy´s gland itself, with diversified form and size according to the species. The Duvernoy´s gland is formed by secretory tubules and excretory ducts. The secretory tubules cells may produce substances of different chemical compositions according to the specie. The histochemical results showed that the majority of the analyzed species present a gland of seromucous type, and only one species has the Duvernoy´s gland classified as mucoserous. The secretion granules present in the cytoplasm of the secretory cells observed in the eletronmicrographics corroborate the histochemical results, since the electron density itself characterizes the cells. The function of the Duvernoy´s gland is directly related to the feeding habit of these serpents and to the way they subjugate preys. The serpents that present the Duvernoy´s gland with mucoserous characteristics subjugate their preys by constriction. On the other hand, the species that have the Duvernoy´s gland of seromucous type use the prey poisoning for feeding. Therefore, the serpents guarantee the success of the immobilization of preys, impeding them to use dangerous defensive tactics against the snake, independently of the use of the constriction.

As serino proteases participam de diversos processos fisiológicos (tal como o de coagulação) e patológicos. Essas enzimas estão amplamente distribuídas entre as espécies, são também toxinas dos venenos de serpentes, sendo denominadas SVSPs (snake venom serine proteases). Essas SVSPs são multifuncionais e contêm uma tríade catalítica formada pelos aminoácidos HDS. Algumas SVSPs são comercialmente disponíveis, sendo indicadas para o tratamento de infarto do miocárdio, tromboses e embolia pulmonar. No veneno de Crotalus durissus terrificus estão descritas até o momento, apenas duas SVSPs sendo que a mais estudada é a giroxina que representa cerca de 2,5% do veneno total. No presente estudo foi reportado a clonagem de sete serino proteases amplificadas a partir de uma biblioteca de cDNA de glândula de veneno de um único espécime adulto de Crotalus durissus terrificus. Estes clones foram analisados com relação à organização do cDNA, estrutura e prováveis funções. A construção do modelo tridimensional da giroxina permitiu verificar as similaridades com tripsina, trombina e outras SVSPs. A glicosilação e a presença de muitas pontes dissulfetos dificultam a obtenção das SVSP recombinantes na forma solúvel e com atividade, por expressão em E.coli. Assim, neste trabalho foi abordada a expressão em células de mamífero (que realiza as modificações pós-traducionais) com resultados promissores. Para tanto, o peptídeo sinal de Igk, a seqüência madura e a região 3 UTR da giroxina foram clonados no vetor pED, originando um novo vetor (pED-Giro). Este vetor carrega o peptídeo sinal de Igk, o que possibilitou a secreção da giroxina para o meio de cultura. O vetor pED-Giro foi transfectado em células CHO DXB11 dhfr e COS-7. A giroxina foi detectada no extrato total das células COS-7 por western blot e, em seguida, purificada do meio de cultura com coluna de afinidade (Benzamidina Sepharose) e demonstrado sua integridade pelo ensaio de atividade esterásica. / The serine proteases affect several physiological processes (such as the coagulation cascade) and pathological ones. These enzymes are widely distributed beyond the species; they are also toxins from snake venoms and are called SVSPs (snake venom serine proteases). These SVSPs are multifunctional and have a catalytic triad formed by HDS amino acids. Some of them are commercially available for use in clinical treatment for heart attack, tromboses and pulmonary embolism. So far, in Crotalus durissus terrificus venom only two SVSPs are described and gyroxin is considered the most studied SVSP which represents about 2,5% of the total venom. In the present study was reported the cloning of seven serine proteases amplified from a cDNA library of a venomous gland of a single adult specimen from Crotalus durissus terrificus venom. These clones have been analyzed in relation to the cDNA organization, structure and probable functions. The three-dimensional model of the gyroxin made possible the analysis of similarities with trypsin, thrombin and other SVSPs. The glycosylation and many disulfide bonds of the SVSPs make difficult the expression in E.coli to obtain the soluble recombinant toxin with activity. The expression in mammalian cells is very promising, because it is possible to make pos translation modification and to obtain the recombinant toxin secreted to the culture medium. The IgK signal peptide, the mature sequence and 3\'UTR region of gyroxin were cloned in the pED expression vector resulting in a new vector (pED-Giro). This vector carries the Igk signal peptide, which allows the secretion of the protein to the culture medium. The pED-Giro vector was transfected in CHO DXB11 dhfr and COS-7 cells. The gyroxin was detected in COS-7 total extract by western blot and after, purified from the medium culture and its integrity was confirmed by esterase activity assay.

Submitted by admin tede (tede@pucgoias.edu.br) on 2016-08-17T14:16:30Z No. of bitstreams: 1 SILVIO JOSE DE QUEIROZ.pdf: 8869081 bytes, checksum: 28d26b617f56db3485bb151beb37b8ad (MD5) / Made available in DSpace on 2016-08-17T14:16:30Z (GMT). No. of bitstreams: 1 SILVIO JOSE DE QUEIROZ.pdf: 8869081 bytes, checksum: 28d26b617f56db3485bb151beb37b8ad (MD5) Previous issue date: 2010-08-09 / Infections caused by multiresistant microorganisms are associated with bacteria acquired in the hospital environment. The selective pressure of antimicrobials is an important factor in the selection and spread of resistance genes in the environment. The use of different molecules produced by living organisms has demonstrated activity against microorganisms. OBJECTIVES: To assess the activity of the crude venom of Bothrops moojeni on gram-negative bacteria producing and non producing metallo- -lactamase and -lactamases with wide spectrum, to identify component (s) of the poison with activity against gram-negative bacteria producing and non producing metallo- -lactamase and -lactamases with wide spectrum and to identify the antibacterial activity of crude venom of the snake B. moojeni in gram negative non-fermentative and fermentative glucose producing different enzymes to degrade -lactams. METHODOLOGY: The poison used was extracted from snake B. moojeni kept on the collection of the Center for Biological Studies and Research (CEPB) Catholic University of Goias Micro-organisms used in this study were obtained from the American Type Culture Collection and Bank of Microorganisms, Laboratory of Clinical Microbiology and MSc in Environmental and Health Sciences at the Catholic University of Goias, including Acinetobacter baumannii (blaIMP2), Pseudomonas aeruginosa (ATCC 27853), Pseudomonas aeruginosa blaIMP1, blaVIM1, blaVIM2, blaSPM1 and producing carbapenemase No. 278 bank) Escherichia coli (ATCC 25922 and ATCC 35 218), Klebisiella pneumoniae (ATCC 700603). The test was conducted with the crude venom applied on disk diffusion in different dilutions. Protein concetration was determined for each venon sample, buy using the Bradford. RESULTS: The microbiological test showed that the venom demonstrated antimicrobial activity, both for producing bacteria and non-producing -lactamase and -lactamases bactérias with wide spectrum and that this activity is probably due to the action of L-amino acid oxidase. CONCLUSIONS: Different concentrations of crude venom of the snake B. moojeni inhibited growth of gram-negative bactéria producing ond non producing metallobeta- lactamase and extende spectrum beta-lactamese ond the size of the allo of growth inhibited diminished when decreased concentration of the crud venon and consequentley the decrese of concentration of protein, demonstrating that the effect of crude venom was dose dependent. / As infecções causadas por microrganismos multirresistentes estão associadas com bactérias adquiridas no meio ambiente hospitalar. A pressão seletiva dos antimicrobianos é um importante fator de seleção e disseminação dos genes de resistência no meio ambiente. O uso de moléculas produzidas por diferentes seres vivos tem demonstrado atividade contra microrganismos. OBJETIVOS: Avaliar a atividade do veneno bruto da Bothrps moojeni sobre bactérias gram-negativas produtoras e não produtoras de metalo- -lactamase e de -lactamases de espectro ampliado, identificar o(s) componente(s) do veneno com atividade contra bactérias gram-negativas produtoras e não produtoras de metalo- -lactamase e -lactamases de espectro ampliado e identificar a atividade antibacteriana do veneno bruto da serpente B. moojeni em bactérias gram negativas fermentadoras e não fermentadoras de glicose produtoras de diferentes enzimas degradadoras de -lactâmicos. METODOLOGIA: O veneno utilizado foi extraído das serpentes B. moojeni, mantidas no serpentário do Centro de Estudos e Pesquisas Biológicas (CEPB) da Pontifícia Universidade Católica de Goiás. Os microrganismos utilizados neste estudo foram obtidos da “American Type Culture Collection” e do Banco de Microrganismos do Laboratório de Microbiologia Clínica e do Mestrado em Ciências Ambientais e Saúde da Pontifícia Universidade Católica de Goiás, sendo Acinetobacter baumannii (blaIMP2), Pseudomonas aeruginosa (ATCC 27853), Pseudomonas aeruginosa blaIMP1, blaVIM1, blaVIM2, blaSPM1 e produtora de carbapenemase banco n° 278) Escherichia. coli (ATCC 25922 e ATCC 35218), Klebisiella pneumoniae (ATCC 700603). O ensaio foi realizado com o veneno bruto aplicado sobre disco de difusão em diferentes diluições. A partir das amostras obtidas, foi realizada a dosagem de proteínas utilizando o método descrito por Bradford. RESULTADOS: O teste microbiológico mostrou que o veneno bruto possui atividade antimicrobiana, tanto para bactérias produtoras como para as não produtoras de metalo- -lactamase e -lactamases de espectro ampliado e que esta atividade se deve possivelmente pela ação da L-aminoácido oxidase. CONCLUSÕES: Diferentes concetrações diluições do veneno bruto da serpente B. moojeni inibiram o crescimento de bactérias gram-negativas produtoras e não produtoras de metalo- -lactamases e -lactamases de espectro ampliado e que o tamanho do alo de inibição do crescimento bacteriano diminuiu na medida em que diminui a concentração do veneno bruto e consequentemente à quantidade de proteínas, demonstrando que o veneno bruto foi dose dependente.

As serino proteases participam de diversos processos fisiológicos (tal como o de coagulação) e patológicos. Essas enzimas estão amplamente distribuídas entre as espécies, são também toxinas dos venenos de serpentes, sendo denominadas SVSPs (snake venom serine proteases). Essas SVSPs são multifuncionais e contêm uma tríade catalítica formada pelos aminoácidos HDS. Algumas SVSPs são comercialmente disponíveis, sendo indicadas para o tratamento de infarto do miocárdio, tromboses e embolia pulmonar. No veneno de Crotalus durissus terrificus estão descritas até o momento, apenas duas SVSPs sendo que a mais estudada é a giroxina que representa cerca de 2,5% do veneno total. No presente estudo foi reportado a clonagem de sete serino proteases amplificadas a partir de uma biblioteca de cDNA de glândula de veneno de um único espécime adulto de Crotalus durissus terrificus. Estes clones foram analisados com relação à organização do cDNA, estrutura e prováveis funções. A construção do modelo tridimensional da giroxina permitiu verificar as similaridades com tripsina, trombina e outras SVSPs. A glicosilação e a presença de muitas pontes dissulfetos dificultam a obtenção das SVSP recombinantes na forma solúvel e com atividade, por expressão em E.coli. Assim, neste trabalho foi abordada a expressão em células de mamífero (que realiza as modificações pós-traducionais) com resultados promissores. Para tanto, o peptídeo sinal de Igk, a seqüência madura e a região 3 UTR da giroxina foram clonados no vetor pED, originando um novo vetor (pED-Giro). Este vetor carrega o peptídeo sinal de Igk, o que possibilitou a secreção da giroxina para o meio de cultura. O vetor pED-Giro foi transfectado em células CHO DXB11 dhfr e COS-7. A giroxina foi detectada no extrato total das células COS-7 por western blot e, em seguida, purificada do meio de cultura com coluna de afinidade (Benzamidina Sepharose) e demonstrado sua integridade pelo ensaio de atividade esterásica. / The serine proteases affect several physiological processes (such as the coagulation cascade) and pathological ones. These enzymes are widely distributed beyond the species; they are also toxins from snake venoms and are called SVSPs (snake venom serine proteases). These SVSPs are multifunctional and have a catalytic triad formed by HDS amino acids. Some of them are commercially available for use in clinical treatment for heart attack, tromboses and pulmonary embolism. So far, in Crotalus durissus terrificus venom only two SVSPs are described and gyroxin is considered the most studied SVSP which represents about 2,5% of the total venom. In the present study was reported the cloning of seven serine proteases amplified from a cDNA library of a venomous gland of a single adult specimen from Crotalus durissus terrificus venom. These clones have been analyzed in relation to the cDNA organization, structure and probable functions. The three-dimensional model of the gyroxin made possible the analysis of similarities with trypsin, thrombin and other SVSPs. The glycosylation and many disulfide bonds of the SVSPs make difficult the expression in E.coli to obtain the soluble recombinant toxin with activity. The expression in mammalian cells is very promising, because it is possible to make pos translation modification and to obtain the recombinant toxin secreted to the culture medium. The IgK signal peptide, the mature sequence and 3\'UTR region of gyroxin were cloned in the pED expression vector resulting in a new vector (pED-Giro). This vector carries the Igk signal peptide, which allows the secretion of the protein to the culture medium. The pED-Giro vector was transfected in CHO DXB11 dhfr and COS-7 cells. The gyroxin was detected in COS-7 total extract by western blot and after, purified from the medium culture and its integrity was confirmed by esterase activity assay.

A reprodução é custosa para ambos os sexos, mas a magnitude dos gastos e sua relação com o sucesso reprodutivo diferem entre os gêneros. Os custos reprodutivos são divididos em duas categorias: custos de sobrevivência e custos energéticos. Crotalus durissus possui um ciclo reprodutivo sazonal com cópula ocorrendo no outono e a parturição no final no verão. Os machos competem por fêmeas receptivas. A inferência de custos reprodutivos associados à sobrevivência foi realizada por meio de levantamentos das taxas de atividade entre machos e fêmeas. A variação da quantidade de gordura abdominal e dos substratos energéticos do fígado e dos rins foi avaliada para mensurar o custo energético. Os resultados sugerem que para fêmeas a reprodução exige um alto investimento energético evidenciado pelos maiores níveis de gordura abdominal e de lipídios no fígado durante a fase vitelogênica e o padrão de atividade diferenciada entre machos e fêmeas sugere que a estação reprodutiva embute um custo de sobrevivência maior para os machos. / Reproduction is costly for both sexes, but the magnitude of spending and its relation to reproductive success differ between genders. Reproductive costs are divided into two categories: survival costs and energy costs. Crotalus durissus has a seasonal reproductive cycle with mating occurring in the fall and parturition in late summer. The males compete for receptive females. The inference of survival costs was accomplished through surveys of activity rates between males and females. The variation of the amount of abdominal fat and energy substrates in liver and kidneys was evaluated to measure the energy cost. The results suggest that for females reproduction requires a high energy investment - evidenced by the higher levels of abdominal fat and lipids in the liver during vitellogenic phase and activity patterns differentiated between males and females suggests that the reproductive season embeds a higher cost of survival for males.