Ecologia de Lithobates catesbeianus (SHAW, 1802) e relações com anfíbios da região de Botucatu, SP (Amphibia, Anura)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Efeito do cloreto de cádmio sobre as gônadas de girinos de rãs-touro (Lithobates catesbeianus)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Atividade de fosfomonohidrolases presentes em fígado de girino de rã-touro (Lithobates catesbeianus) durante a metamorfose

Pós-graduação em Biotecnologia - IQ

Atividade de enzimas relacionadas ao processo de ossificação em girinos de Lithobates catesbeianus

Pós-graduação em Biotecnologia - IQ

Median lethal concentration of formaldehyde and its genotoxic potential in bullfrog tadpoles (Lithobates catesbeianus)

In order to avoid that contaminated frog farms animals escaping in the environment and become potential vector of emergent diseases, studies with disinfection protocol are strictly necessary. The formaldehyde is one of the compounds tested in fungal disinfection protocols and also used in aquaculture. This study aimed to determine the median lethal concentration (LC50-96h) of formaldehyde in bullfrog tadpoles and to evaluate the possible genotoxic effects in acute exposition. Accordingly, the animals were exposed to formaldehyde in the concentrations of 6, 9, 12, 15, and 18 mg L(-1), and after 96 h blood samples were drawn for the micronucleus (MN) test. The LC50-96h was 10.53 mg L(-1), and the MN frequency increased in proportion to the formaldehyde concentrations, with an estimated frequency in the negative control being 1.35 MN/individual. We concluded that formaldehyde is genotoxic to tadpoles of bullfrogs in the tested concentrations, and the choice of this chemical should be contemplated before its use in animals in captivity.

Isolamento e resistência a antimicrobianos de cepas de Streptococcus spp. provenientes de rãs-touro (Lithobates catesbeianus)

Twelve bullfrogs were selected from two commercial frog farms and clinically diagnosed as attacked by Streptococcus disease. Sixty samples were collected, and Streptococcus spp. was isolated from all bullfrog, being 12 (100%) from the encephalus, seven from the kidneys (58.3%), three from the liver (25%), two from the spleen (16.6%), and one from the ascitic liquid (8.3%). Streptococcus -hemolytic were isolated from all the 60 samples, which were sensible to chloramphenicol (100%), gentamycin (100%), vancomycin (96%), cefotaxime (96%), and cefoxitine (92%).

Duration of spermatogenesis in the bullfrog (Lithobates catesbeianus)

The bullfrog (Lithobates catesbeianus) has substantial economic importance and has also been used as an experimental model for biological studies in the fields of pharmcology, medicine, and reproductive biology, especially studies addressing gametogenesis. However, there is a lack of comprehensive information in the literature regarding testis structure and function in this amphibian. The main objective of the current study was to estimate the duration of the various phases of spermatogenesis in this vertebrate. Sixteen sexually mature bullfrogs received an intracoelomic administration of tritiated thymidine. Testes were analyzed at various times between I h and 33 d after administration to detect the most advanced germ cell types labeled at each interval, as well as labeled preleptotene spermatocytes, which presumably originated from spermatogonial stem cells. The duration of the spermatogonial, spermatocytic, and spermiogenic phases of spermatogenesis in the bullfrog were approximately 18, 14, and 8 d, respectively. Thus, the total duration of the spermatogenesis process from early spermatogonia through to spermatozoa was 40 d in this species, similar to that of most previously investigated mammalian species. To our knowledge, this is the first reliable report on the duration of the full spermatogenic process in any amphibian species. These findings will be very useful for tracking the pace of germ cells in studies involving spermatogonial transplantation in lower vertebrates. (C) 2009 Elsevier B.V. All rights reserved.

Profile of cortisol, glycaemia, and blood parameters of American Bullfrog tadpoles Lithobates catesbeianus exposed to density and hypoxia stressors

O objetivo deste estudo foi avaliar a resposta fisiológica (cortisol, glicemia e parâmetros sanguineos) de girinos de rã-touro (Lithobates catesbeianus) em diferentes densidades e após exposição aérea. Os animais utilizados no experimento estavam entre os estágios 31 a 39, na fase de pró-metamorfose sendo testados 1 girino/L (Tratamento 1), 5 girinos/L (Tratamento 2) e 10 girinos/L (Tratamento 3), conduzidos em 3 réplicas simultâneas durante 12 dias. O sangue foi retirado por rompimento do vaso caudal na condição de Normóxia - N (tempo zero) e Hipóxia - H (tempo de 15 minutos de exposição ao ar). Foi observado um aumento nos valores de cortisol, aos 4 e 8 dias de exposição aérea retornando aos valores basais ao final do experimento, apesar de não haver diferenças significativas. A glicemia não apresentou diferenças significativas quanto aos estressores aplicados. Os parâmetros hematológicos da série branca, principalmente, o número de linfócitos, neutrófilos e eosinófilos mostraram diferença significativa aos 12 dias de experimentação quando comparados com o momento zero; concluindo-se que, em condições controladas, o adensamento de até 10 girinos/litro e a exposição aérea por 15 minutos não apresentou danos aos girinos de rã-touro durante o período experimental. O padrão de resposta a estes estímulos talvez seja expresso em outro nível hormonal (corticosterona).

Tecido hepático e corpo adiposo de rã-touro (Lithobates catesbeianus) submetido a diferentes fotoperíodos

The study was conducted to evaluate the liver tissue weight, body fat weight, the fatty somatic index and the liver somatic index of bullfrogs subjected to three photoperiod during the growing phase (initial 30 days) and finishing (60 days). The treatments were 16 h light and 8 h dark, 12 h of light and dark and 8 h light and 16 h dark. The experimental design was completely randomized design with three replications. We used 36 bullfrogs/m2 weighing from 22 to 28 g. The results were submitted to ANOVA and Tukey Test at 5% probability. Differences were found for the weight of body fat, liver tissue weight, liver somatic index and fatty somatic index the bullfrog to the treatments and the study periods and there is greater accumulation of energy to ninety days. The photoperiod did not influence body weight and fat weight of the liver tissue of bullfrog at the end of the fattening period and the difference fatty somatic index and liver somatic index were influenced by body weight of animals.

Skin bradykinin-related peptides (BRPs) and their biosynthetic precusors (kininogens): comparisons between various taxa of Chinese and North American ranid frogs.

Bradykinins and related peptides (BRPs) occur in the defensive skin secretions of many amphibians. Here we report the structures of BRPs and their corresponding biosynthetic precursor cDNAs from the Chinese brown frog, Rana chensinensis, and the North American leopard frog, Lithobates pipiens. R. chensinensis skin contained four transcripts each encoding a different kininogen whose organizations and spectrum of encoded BRPs were similar to those reported for the pickerel frog, Lithobates palustris. In contrast, from L. pipiens, a single skin kininogen was cloned whose structural organization and spectrum of mature BRPs were similar to those reported for the Chinese piebald odorous frog, Huia schmackeri. These data also implied that the endogenous precursor processing proteases in each species pair have identical site-directed specificities, which in part may be dictated by the primary structures of encoded BRPs. Thus the spectra of skin BRPs and the organization of their biosynthetic precursors are not consistent with recent taxonomy. The natural selective pressures that mould the primary structures of amphibian skin secretion peptides are thought to be related to the spectrum of predators encountered within their habitats. Thus similarities and differences in skin bradykinins may be reflective of predator spectra rather than indicative of species relatedness.

Characterizing the impact of multiple potential enemies (predators and parasites) on the behaviour of ranid tadpoles

In order to fully understand an organism's behaviours the interactions between multiple enemies or selective pressures need to be considered, as these interactions are usually far more complex than the simple addition of their effects in isolation. In this thesis, I consider the impact of multiple enemies (fish predators and parasites) on the behaviour of three larval anurans (Lithobates sylvaticus, L. clamitans and L. catesbeianus). I also determine whether species that differ in life-histories and habitat preferences possess different antipredator mechanisms and how this affects species responses to multiple enemies. I show that the three Ranid larvae respond differently to the trade-off imposed by the presence of both fish predators and trematode parasites within the environment. The two more permanent pond breeders (L. clamitans and L. catesbeianus) increased activity when in the combined presence of predators and parasites. In contrast, the temporary pond breeder (L. sylvaticus) decreased activity in the combined presence of predator and parasites, in the same manner as they responded to fish alone. Further, the presence of fish along with parasites increased the susceptibility of both L. sylvaticus and L. clamitans to trematode infection, whereas parasite infection in L. catesbeianus was unaffected by the presence of fish. A second experiment to assess palatability of the three anuran species to fish, revealed a range of palatabilities, with L. catesbeianus being least palatable, L. clamitans being somewhat unpalatable, and L. sylvaticus being highly palatable. This result helps to explain the species differences in tthe observed behaviour to the combined presence of fish and parasites. In conclusion, the results from this study highlight the importance of considering multiple selective pressures faced by organisms and how this shapes their behaviour.

Le roseau commun (Phragmites Australis) influence-t-il la composition spécifique et le développement larvaire d'amphibiens?

Les plantes envahissantes sont considérées comme l'une des plus grandes menaces sur les espèces, mais leurs impacts sur les amphibiens demeurent peu connus. L’objectif de ce projet est de déterminer l’effet de l’établissement du roseau commun (Phragmites australis) sur la répartition des amphibiens et sur leur développement larvaire. Il est présumé que cette plante s'accapare l'espace et les ressources disponibles en produisant une importante biomasse, qu’elle peut modifier l’hydrologie des marais et la structure des communautés d’amphibiens. J’ai évalué les facteurs influençant la répartition des amphibiens selon les caractéristiques des étangs et du paysage dans 50 sites envahis ou non à divers degrés. Des expériences ont également été menées afin d’étudier les répercussions de trois densités de roseau sur des têtards de la Grenouille des bois (Lithobates sylvaticus) et sur la qualité de son habitat. Mes résultats suggèrent que le roseau à forte densité ralentit le développement larvaire de la grenouille des bois et influence les assemblages phytoplanctoniques. Cependant, il n’y a aucune relation entre la densité de la plante et la survie, la morphologie des têtards et les caractéristiques biotiques et abiotiques de l’eau. Dans notre aire d’étude, le paysage autour des étangs a une plus grande influence sur la répartition des amphibiens que l'établissement du roseau. Toutefois, la probabilité d’assèchement est plus élevée lorsque la plante est établie en grande quantité, ce qui, si l’envahissement s’intensifie, aura un effet néfaste sur la survie des têtards et mettra en péril la persistance des populations.

Physiological response of American bullfrog tadpoles to stressor conditions of capture and hypoxia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Modelo de avaliação de risco : São o crescimento e a estequiometria dos girinos afetados pelo efeito interativo entre a presença do predador e da densidade de coespecíficos?

Many prey organisms change their phenotype to reduce the predation risk. However, such changes are associated with trade-offs, and can have negative effects on prey growth or reproduction. Understand how preys assess the predation risk is essential to evaluate the adaptive value of predator-induced phenotypic and its ecological consequences. In this study, we performed a mesocosm experiment to test: i) if growth and stoichiometry of Lithobates catesbeianus tadpoles is altered in response to giant water bug presence (Belostoma spp.); ii) if these responses depend on tadpoles density in environment. Here, we show that tadpoles growth and stoichiometry are not changed by predator presence, neither in low nor in high densities. Our results suggest that tadpoles exposed to predation risk regulate their physiology to preserve the elemental stoichiometric homeostase of their body and excretion. Further, point out to need for future studies that elucidate under what conditions growth and stoichiometry are changed in response to predation risk

Time interval from ovulation to extrusion in female bullfrog in different photoperiods

It was analyzed in this work the influence of photoperiod on time interval from ovulation induction period to extrusion of ovocits in female bullfrogs (Lithobates catesbeianus). It was used 54 females reared from metamorphosis to 9 months of age under three photoperiods: dark time (DL 0:24), 16 hours of daylight (DL 16:8) and 12 hours of daylight (DL 12:12). Ovulation was induced by intramuscular application of two doses of LHRHa with 12 hours of interval between the injections. After 10, 25, 28, 31, 34 and 37 hours from the first hormone injection, 10-gram samples (3,000 eggs) were extracted from each female at each time interval and fertilized. Egg hatching rate was checked in each sample 72 hours after fertilization. Analysis of variance showed a significant effect of extrusion delay and the interaction between photoperiod and this delay. Extrusion should be carried out 33, 24 and 26 hours after the first hormone dosage in females reared in environments without light, with 12 hours of daylight and with 16 hours of daylight, respectively, to obtain the maximum fertilization rate.