Orientador: Luciana Bolsoni Lourenço / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-27T00:43:09Z (GMT). No. of bitstreams: 1 Seger_KarinRegina_M.pdf: 1961069 bytes, checksum: ac950c9cf223e51f3374d9378d583572 (MD5) Previous issue date: 2014 / Resumo: O gênero Dendropsophus apresenta uma grande diversidade de espécies, compreendendo atualmente mais de 90 espécies de anuros hilídeos, e se caracteriza por apresentar como principal sinapomorfia o número cromossômico 2n=30. Muitas problemáticas taxonômicas ainda cercam esse grupo, como a referente a Dendropsophus nanus e Dendropsophus walfordi. Tais espécies já foram consideradas sinônimas com base em dados morfológicos, proposta que foi refutada após análise de canto. Recentes inferências filogenéticas têm apontado o possível parafiletismo dessas duas espécies de anuros, mas nenhuma delas teve a amostragem devidamente desenhada para avaliar cuidadosamente a questão taxonômica decorrente dessa observação. Mesmo o estudo filogenético que incluiu o maior número de representantes dessas espécies contou com apenas cerca de 700 pb de um único marcador genético (gene ribossomal mitocondrial 12S). Além disso, não incluiu nenhum exemplar da localidade-tipo de ambas as espécies e, embora a amostra dessa espécie tenha sido a maior utilizada até o momento em análises filogenéticas, ainda não foi suficiente para cobrir toda a grande área de ocorrência desses anuros. Com o intuito de solucionar tais problemas e contribuir para a análise da delimitação dessas espécies, foi investigado o possível parafiletismo entre as espécies D. nanus e D. walfordi, por meio da comparação de sequências dos genes mitocondriais 12S, 16S e CO1, e com a amostragem de indivíduos de 24 localidades abrangendo a maior parte da distribuição incluindo as localidades-tipo de cada uma das duas espécies. Foi observada alta estruturação genética do grupo de interesse pela análise de variância molecular (AMOVA) e cinco principais clados (A-E) foram identificados em todos os cladogramas inferidos. O parafiletismo de D. nanus com relação a D. walfordi foi observado em todas as análises filogenéticas. As distâncias genéticas entre os clados A-E foram altas e permitem sugerir que o grupo em questão represente na realidade um complexo com mais de duas espécies diferentes / Abstract: The Dendropsophus genus presents a great diversity of species, currently including more than 90 species of hylid frogs, and its main synapomorphy is the chromosome number 2n = 30. Many taxonomic problems still surround this group, as those referred to Dendropsophus nanus and Dendropsophus walfordi. Such species have been considered synonymous based on morphological data, but that proposal was rejected after call analysis. Recent phylogenetic inferences have pointed out the possible paraphyly of these species of frogs, but they were not designed to carefully evaluate the taxonomic question arising from this observation. Even the phylogenetic study that included a large number of representatives of these species did not sampled their type locality and was based on only ~ 700 bp of a single genetic marker (mitochondrial 12S ribosomal gene). In order to solve these problems and contribute to the analysis of these species delimitation, we investigated the possible paraphyly between D. nanus and D. walfordi by comparing the sequences of the mitochondrial genes 12S, 16S and CO1, and sampling specimens from 24 localities, including the type-locality of each of these species. High genetic structuration was observed inside this group by the analysis of molecular variance (AMOVA) and five major clades (A-E) were recognized in all inferred cladograms. The paraphyly of D. nanus with respect to D. walfordi was observed in all phylogenetic analyses. The genetic distances among the Clades A-E were high and allowed to suggest that in fact a complex with more than two different species exist in the analyzed group / Mestrado / Biodiversidade Animal / Mestra em Biologia Animal

A aquicultura é apontada como um mercado estratégico para o desenvolvimento sustentável, produção de alimentos e ampliação de fronteiras inexploradas no Brasil. No entanto, como outros sistemas de produção animal, este setor enfrenta problemas com doenças resultantes de sua intensificação, como os aspectos sanitários da produção e a falta de estrutura para o diagnóstico das principais enfermidades infecciosas. Durante os últimos 20 anos, os vírus da família Iridoviridae, em especial membros do gênero Ranavirus, têm sido responsáveis por epizootias de grande impacto ecológico e econômico, envolvendo um grande número de espécies de peixes, anfíbios e répteis de importância na aquicultura de várias partes do mundo. No entanto, as informações sobre a ocorrência de infecções de peixes e anfíbios causadas por ranavírus no Brasil são limitadas. O presente projeto compreendeu o isolamento em cultivo celular de estirpe de Frog Virus 3-símile, no Estado de São Paulo, confirmada por sequenciamento nucleotídico do gene MCP e subsequente RFLP, assim como caracterização fenotípica e de cinética de replicação do isolado. Amostras de fígado, baço e rins de rãs-touro gigante provenientes de ranário comercial, positivas ao diagnóstico molecular para Ranavirus, foram utilizadas para isolamento viral em cultivo celular. Efeito citopático foi detectado na segunda passagem em células BF-2 e posterior confirmação do isolamento foi feita por PCR e RFLP. A caracterização fenotípica viral foi feita com microscopia eletrônica de transmissão, a qual permitiu a confirmação de morfologia semelhante às partículas de Ranavirus, além da realização de curva de replicação, indicando maiores títulos virais no quarto dia após inoculação. Também foi feito teste de susceptibilidade a solventes, que confirmou a presença de partículas envelopadas. Os resultados obtidos nesse estudo poderão contribuir para a futura compreensão da biologia dos iridovírus circulantes como agentes etiológicos de ranaviroses em anfíbios no Estado de São Paulo. / Aquaculture is appointed as a strategic market for sustainable development, food production and expansion of unexplored frontiers in Brazil. However, just like other animal production systems, this area faces problems due its intensification, like the production sanitation aspects and the lack of structure for the diagnosis of major infectious diseases. During the last 20 years, viruses from Iridoviridae family, especially Ranavirus genera, have been responsible for major economic and ecological impactant epizootic diseases in a great number of fish, amphibian and reptile species in many parts of the world. However, information on the occurrence of infections in fishes and amphibians caused by Ranavirus in Brazil are limited. In this context, this project aimed to isolate a Frog Virus 3-like strain detected in São Paulo state in cell culture, which was later confirmed by nucleotide sequencing of MCP gene and further RLFP technique and also phenotipical characterization and replication kinetics of the obtained strain. Liver, spleen and kidney samples from american bullfrogs from commercial frog ponds, positive by molecular diagnostic for Ranavirus, were used for viral isolation in cell culture. Citopathic effect was detected during the second passage in cells and later confirmed by PCR and RFLP. The viral characterization was carried out with transmission electronic microscopy, which confirmed similar morphology of viral particles to those of Ranavirus, besides the construction of a growth curve which indicated larger titres on the fourth day post inoculation. A test for solvent sensitivity was also performed and confirmed the presence of enveloped particles. The results obtained in this study will contribute to the understanding of the biology of the circulating iridovirus in São Paulo state as an etiological agent of Ranavirus infection in amphibians.

O objetivo deste estudo foi coletar e reunir informações sobre a diversidade de anfíbios e répteis nas bacias hidrográficas do Alto e Médio Paranapanema, estado de São Paulo e avaliar os impactos que o pastoreio de gado pode provocar na riqueza e abundância de anuros e lagartos de formações abertas de Cerrado. A riqueza de anuros registrada no Alto Paranapanema é muito superior a encontrada no Médio Paranapanema, existindo um gradiente decrescente na riqueza de espécies partindo da porção sul e sudeste do Alto Paranapanema, onde predominam a floresta ombrófila, em direção ao Médio Paranapanema que apresenta formações de Cerrado e floresta estacional. O pastoreio de gado não causou efeitos negativos na riqueza e abundância total dos anuros e lagartos amostrados. É possível que a intensidade de pastoreio ocorrida nestas formações não foi suficiente para promover modificações consideráveis na estrutura das fitofisionomias, o que pode ter minimizado alterações nos nichos ecológicos das espécies. / The aim of this study is to collect information on the diversity of amphibians and reptiles in the hydrographic basins of the Upper and Middle Paranapanema, São Paulo state and assess the impact that cattle grazing can provoke on the species richness and abundance of anurans and lizards of open Cerrado formations. The species richness of anurans reported in the Upper Paranapanema is far greater to that found in the Middle Paranapanema, with a decreasing gradient in the richness from the southern and southeastern portion of Upper Paranapanema, where the ombrophilous forest predominates, towards the Middle Paranapanema in which Cerrado formations and seasonal forests are dominant. Cattle grazing did not cause negative effects on the species richness and total abundance of the anurans and lizards sampled. It is possible that the intensity of grazing was insufficient to cause significant changes in the structure of the phytophysiognomies, which could have minimized alterations in the ecological niches of the species.

Orientador: Denis Otavio Vieira de Andrade / Banca: José Eduardo de Carvalho / Banca: Kênia Cardoso Bicego / Abstract: As ectotherms, amphibians may experience wide fluctuations in body temperature and, due to their high skin permeability, terrestrial species face a constant risk of desiccation. These two variables (temperature and dehydration) are centrally relevant for water balance regulation because they directly affect water flux through the integument, i.e. skin evaporative water loss (EWLSkin) and water uptake (WU). In addition, as nearly all anurans are lung breathers, the respiratory water loss (EWLResp) will add to EWLSkin. Although the contribution of EWLResp to total EWL (EWLTotal) is generally assumed to be negligible, the partitioning between EWLSkin and EWLResp varies among species and is affected by temperature and dehydration. Therefore, we investigated the combined effects of temperature and dehydration on the rates of EWL through the skin and the lungs of the terrestrial toad, Rhinella schneideri. Subsequently, we evaluated how water uptake (WU) through the pelvis skin was affected by temperature and dehydration. To this aim, we measured rates of EWLTotal and EWLSkin in intact and masked adult toads at 15, 25 and 35 oC under fully hydrated condition and dehydrated until they have lost 10% and 20% of their initial body mass. Masked toads were able to breath normally during the measurement of EWLSkin; EWLResp was calculated as EWLTotal minus EWLSkin. Rates of EWL were also determined using biophysical agar models of R. schneideri specimens, which allowed the estimation of ski... (Complete abstract click electronic access below) / Resumo: Como animais ectotérmicos, os anfíbios experimentam longas flutuações na temperatura corpórea e, devido à sua alta permeabilidade cutânea, eles também enfrentam o risco constante da dessecação. Essas duas variáveis (temperatura e desidratação) são muito relevantes no contexto do balanço hídrico dado que elas afetam diretamente o fluxo de água através da pele, ou seja, a perda evaporativa de água (PEAPele) e a absorção de água (RE). Além disso, dado que a maioria dos anuros possuem respiração pulmonar, a perda de água respiratória (PEAResp) irá se adicionar à cutânea. Embora a contribuição da PEAResp para a perda de água total (PEATotal) seja geralmente considerada insignificante, a divisão entre PEAPele e PEAResp varia de acordo com as espécies e é afetada pela temperatura e desidratação. Portanto, nós investigamos os efeitos combinados da temperatura e desidratação nas taxas de PEA através da pele e pulmões no sapo terrestre, Rhinella schneideri. Posteriormente, avaliamos como a absorção de água através da pele ventral (RE) foi afetada pela temperatura e desidratação. Para isso, medimos as taxas de PEATotal e PEAPele em sapos adultos intactos e mascarados à 15, 25 e 35 oC e desidratados até perderem 10% e 20% da massa corpórea inicial. Os sapos mascarados foram capazes de respirar normalmente durante a medição da PEAPele; PEAResp foi calculado como PEATotal menos PEAPele. As taxas de perda de agua também foram determinadas usando modelos de ágar de espécimes de R. schneideri... (Resumo completo, clicar acesso eletrônico abaixo) / Mestre

Agriculture ditch management (i.e. removal of vegetation and/or sediments) may disturb native wildlife, such as amphibian bioindicator species. The objective of this thesis was to determine whether ditch management affected northern leopard frogs (Lithobates pipiens). Twelve embryo and tadpole health endpoints were compared in vegetated and managed (treeless or dredged) ditches using in situ cages in 2018 and 2019, while water quality was monitored. The managed treatment did not negatively affect frog health compared to the vegetated treatment. The significantly faster development and larger body size due to warmer water temperatures at the managed treatment were likely advantageous. The water quality was typical of the region regardless of the ditch treatment and the pesticide mixture detected had limited toxicity. The main effects of the studied ditch managements on resident L. pipiens (and potentially other amphibians) appeared sub-lethal, likely beneficial, and mainly the result of temperature differences. Although poorer embryo health was observed at the vegetated treatment, it was likely unrelated to ditch management and possibly due to high specific conductivity causing premature hatching and mortality. Ditch management may be less important for frog health than the surrounding land use, and efforts should be directed at increasing suitable habitat for amphibians in agroecosystems. La gestion des fossés agricoles (plus spécifiquement l'élimination de la végétation des bandes riveraines et /ou le dragage des sédiments) peut perturber la faune indigène, comme les espèces bio-indicateurs d'amphibiens. L'objectif de cette thèse était de déterminer si la gestion des fossés affectait la grenouille léopard du Nord (Lithobates pipiens). Douze paramètres d'évaluation de la santé des embryons et des têtards ont été comparés dans des fossés végétalisés et gérés (sans arbres ou dragués) à l'aide de cages expérimentales in situ en 2018 et 2019, pendant que la qualité de l'eau a été suivie. Le traitement géré n'a pas eu d'effets négatifs sur la santé des grenouilles par rapport au traitement végétalisé. Le développement beaucoup plus rapide et la taille corporelle plus grande en raison des températures plus chaudes de l'eau lors du traitement géré étaient probablement avantageux. La qualité de l'eau était typique de la région quel que soit le traitement du fossé et le mélange de pesticides détecté avait une toxicité limitée. Les principaux effets de la gestion des fossés étudiés sur L. pipiens résidents (et potentiellement d'autres amphibiens) semblaient sublétaux, probablement bénéfiques, et résultaient principalement de différences de température. Bien que la santé des embryons ait été moins bonne lors du traitement végétalisé, elle n'était probablement pas liée à la gestion des fossés et pourrait être due à une conductivité spécifique élevée entraînant une éclosion prématurée et la mortalité. La gestion des fossés peut être moins importante pour la santé des grenouilles que l'utilisation des terres environnantes, et les efforts de protection devraient être dirigés vers une augmentation générale de l'habitat approprié pour les amphibiens dans les agroécosystèmes.

No description available.

To understand present patterns of biodiversity, knowledge of a lineage’s past – both evolutionary and geographic – is required. Here I present the first comprehensive phylogenomic study of an Amazonian poison frog genus, Ameerega, as well as the introduction of a new method for characterizing ancestral distributions via phylogenetic niche modeling, which I use to investigate Ameerega’s biogeographic past. I sequenced thousands of ultraconserved elements from over 100 tissue samples, representing almost every described Ameerega species, as well as undescribed cryptic diversity. My phylogenetic inference diverged strongly from those of previous studies. I also introduce a new phylogenetic niche modeling method, which accounts for issues of bias in other methods by incorporating knowledge of evolutionary relationships into niche models. Given modern-day and paleoclimatic data, species occurrence data, and a time-calibrated phylogeny, my method constructs niche models for each extant taxon, uses ancestral character estimation to reconstruct ancestral niche models, and projects these models into paleoclimate data to provide a historical estimate of the geographic range of a lineage. I demonstrate my method on the Ameerega bassleri group. I also use simulations to show that my method can reliably reconstruct the niche of a known ancestor in both geographic and environmental space.

Orientador: Classius de Oliveira / Banca: Rodrigo Zieri / Banca: Monica Jones Costa / Banca: Rafaela Maria Moresco / Banca: Vitor Hugo Mendonça do Prado / Resumo: A crescente demanda para produções agrícolas e o consequente uso de agrotóxicos nas lavouras, vem se tornando foco das atenções sobre o uso e as conseqüências destes contaminantes químicos ambientais. Utilizando-se de anuros coletados em ambientes agrícolas e em uma Unidade de Conservação, o Parque Nacional das Emas (PNE), ambos região Sudoeste do Estado de Goiás, investigou-se a influência de contaminantes de origem agrícola nos aspectos anatomorfológicos em larvas de anuros bem como as possíveis alterações genótoxicas destes compostos. Para isto, foram coletadas larvas de anuros de 21 espécies de área agrícola e nove da Unidade de Conservação, em diferentes ambientes. Foram realizadas análises de malformações no padrão morfológico (27 espécies), bem como análises de alterações eritrocitárias em três espécies. Os resultados demonstraram alta porcentagem de anormalidades em anuros provenientes de agroecossistemas, quando comparados aos de ambiente natural. Entretanto, as análises de alterações eritrocitárias nucleares nas larvas não mostram significância para a formação de micronúcleo em girinos entre os ambientes estudados. No entanto foi registrada uma maior frequência de células reniformes em girinos de ambiente antropizado. Portanto, nosso estudo aponta que girinos associados a ambientes agrícolas são mais suscetíveis a algum tipo de alteração morfológica, o que reforça a importância da preservação de áreas originais e ainda um olhar mais critico quanto ao uso de... / Abstract: The growing demand for agricultural production and the consequent use of agrochemicals in crops has become a focus of attention on the use and consequences of these environmental chemical contaminants. Using anurans collected in agricultural environments and in a Conservation Unit, the Emas National Park (PNE), both Southwest region of the State of Goiás, investigated the influence of contaminants of agricultural origin on the anatomorphological aspects in anuran larvae as well as the possible genotoxic changes of these compounds. For this, anuran larvae were collected from 21 species of agricultural area and nine from the Conservation Unit, in different environments. Analyzes of malformations in the morphological pattern (27 species) were carried out, as well as analyzes of erythrocyte alterations in three species. The results showed a high percentage of anurans abnormalities from agroecosystems when compared to those from the natural environment. However, the analyzes of nuclear erythrocyte alterations in the larvae do not show significance for the formation of micronucleus in tadpoles between the studied environments. However, a higher frequency of reniform cells was recorded in anthropic environment tadpoles. Therefore, our study indicates that tadpoles associated with agricultural environments are more susceptible to some type of morphological alteration, which reinforces the importance of the preservation of original areas and a more critical view regarding the use of ... / Doutor

The importance of hypovolemia in dehydrational death was assessed in two anuran species. Xenopus laeyis, a species which experiences a significant reduction in circulating plasma volume with dehydration, was used to evaluate the role of sympathetic reflex compensation in hypovolemia. Adrenergic blockade with propranolol or phenoxybenzamine produced no significant reduction in dehydration tolerance in this species, although β-blockade with propranolol appeared to have a minor effect. The role of hypovolemic shock in the terminal circulatory collapse that precedes death in dehydrating anurans was investigated in the toad, Bufo marinus. The activity of lysosomal proteinases (cathepsins) was used as an indicator of the shock state. An assay was developed for the determination of cathepsin activity in toad plasma and was used to demonstrate the presence of proteinases similar in pH dependence to mammalian cathepsins Bl and D. Plasma cathepsin D-type activity increased with dehydration in toads but was not significantly different from the activity in controls or in toads subjected to splanchnic artery occlusion. Toads appear to maintain plasma volume to 20% water loss at the expense of other extracellular fluids. These results do not support the hypothesis that hypovolemia is a direct cause of dehydrational death in anurans.

Aquatic habitats are closely linked to surrounding terrestrial environments via reciprocal subsidies. Much of the research on these subsidies has been focused on streams, while subsidies across aquatic-terrestrial boundaries of other aquatic systems, like temporary ponds, have received little attention. Temporary ponds are seasonally inundated aquatic habitats and patterns of energy transport across transient aquatic-terrestrial boundaries are not well understood. To address the lack of information regarding these cross-habitat subsidies, I quantified: 1) leaf litter inputs, 2) amphibian egg inputs, 3) terrestrial insect inputs, 4) amphibian metamorph emergence, and 5) aquatic insect emergence for eight temporary ponds. Terrestrial insect inputs to ponds represented substantial high-quality subsidies to ponds that are generally unaccounted for in similar studies. While larger ponds produced greater total fluxes to terrestrial habitats, smaller ponds were often more productive per unit area. Therefore, a mosaic of small ponds may produce greater or equivalent subsidies and may be of greater value to terrestrial food webs than a single large pond. Resource subsidies of energy and nutrients can be transported via physical forces or biotic processes, such as animal migration or emigration. I quantified net nutrient fluxes associated with amphibian migrations across aquatic-terrestrial boundaries of eight temporary ponds in Illinois. I measured carbon (C), nitrogen (N), and phosphorus (P) subsidies from forests to ponds, in the form of eggs, and the reciprocal subsidies from ponds to forests via juvenile emigration. Juvenile emergence biomass did not resemble egg biomass, as not all species bred successfully in each pond, resulting in variability in the magnitude of nutrient fluxes both across ponds and species. The forest was not always the recipient system of net nutrient fluxes. Hydroperiod, trophic interactions, and species composition explained some dynamics of N and P subsidies. Most studies have focused on the magnitude of cross-boundary fluxes, but the impact of a subsidy is mediated by the size of the flux as well as the quality and relative abundance of similar resources in the recipient habitat. Long-chain polyunsaturated fatty acids (LC-PUFAs) are necessary for proper physiological function, are unevenly distributed across the landscape, and animals differ greatly in their ability to synthesize them de novo. This creates the potential for limitation and increases their possible importance as a subsidy. I examined LC-PUFA tissue concentration and export in eight species of emerging amphibian metamorphs across eight temporary ponds in a wetland complex. I found that tissue concentrations and export of LC-PUFAs varied across species, but were generally within the ranges of several freshwater fish and aquatic insects. Anurans exported higher amounts of LC-PUFAs than salamanders, largely due to the higher emergence biomass of anurans. Further, I explored fatty acid profiles and immune function of wolf spiders in wetland and upland habitats. Wetland spiders had higher tissue levels of aquatic-derived LC-PUFAs and elevated immune function compared to upland spiders. These patterns suggest aquatically derived nutrients, such as LC-PUFAs, may benefit organisms exploiting freshwater habitats.