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

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

Infestação experimental em girinos de Rana catesbeiana Shaw por copepoditos de Lernaea cyprinacea Linnaeus (Copepoda, Lernaeidae)

The Lernaea cyprinacea Linnaeus, 1758 copepodids experimental contamination on the Rana catesbeiana Shaw, 1802 ladpoles was analized. Sixty percent (12) of infested tadpoles was found parasitized. High mortality range, lower appetence, equilibrium loss and apathy were observed. The parasites showed the preference for the mouth and cloaca: in 72 hours lhe egg saes had already developed.

Effect of oxytocin on transepithelial transport of water and Na+ in distinct ventral regions of frog skin (Rana catesbeiana)

Thoracic, abdominal, and pelvic fragments of ventral skin of Rana catesbeiana were analysed regarding the effect of oxytocin on: (1) transepithelial water transport; (2) short-circuit current, (3) skin conductance and electrical potential difference; (4) Na+ conductance, the electromotive force of the Nat transport mechanism, and shunt conductance; (5) short-circuit current responses to fast Na+ by K+ replacement in the outer compartment, and (6) epithelial microstructure. Unstimulated water and Na+ permeabilities were low along the ventral skin. Hydrosmotic and natriferic responses to oxytocin increased from thorax to pelvis, Unstimulated Na+ conductance was greater in pelvis than in abdomen, the other electrical parameters being essentially similar in both skin fragments. Contribution of shunt conductance to total skin conductance was higher in abdominal than in pelvic skin. Oxytocin-induced increases of total skin conductance, Na+ conductance, and shunt conductance in pelvis were significantly larger than in abdomen, An oscillatory behaviour of the short-circuit current was observed only in oxytocin-treated pelvic skins. Decrease of epithelial thickness and increase of mitochondria-rich cell number were observed from thorax to pelvis, Oxytocin-induced increases of interspaces were more conspicuous in pelvis and abdomen than in thorax.

Avaliação dos fatores ambientais em baias com cobertura de polietileno sobre o desempenho da rã-touro (Rana catesbeiana)

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

Recria de Rã touro (Rana catesbeiana) em tanques-rede alojados em viveiros de tilápia

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

Estufas climatizadas para experimentos ambientais com rãs, em gaiolas

Foram construídas seis estufas climatizadas, instaladas inicialmente no Ranário Experimental da Universidade Federal de Viçosa (UFV) e, posteriormente, no Ranário Experimental da Fundação Universidade Federal do Rio Grande, com o objetivo de realizar experimentos para avaliar os efeitos do ambiente sobre o desempenho de rãs em gaiolas de fibra de vidro. Ambientes com temperaturas de 25ºC e fotoperíodo de 12/12 horas de luz/horas de escuridão (h L/E) serviram para adaptação das rãs por 15 dias antes de cada experimento. Os tratamentos consistiram em simular ambientes com temperaturas variando de 20 a 35ºC e fotoperíodos de 8/16, 12/12 e 16/8 h L/E. Foram realizados experimentos com rã-touro (Rana catesbeiana Shaw, 1802) e rã-manteiga (Leptodactylus ocellatus Linnaeus, 1758). Nessas estufas foi possível estimar que: a) os maiores ganhos de peso de rã-touro foram obtidos entre 27,6 e 29,7ºC, com melhor crescimento entre 28,2 e 30,1ºC; para rã-manteiga os melhores ganhos e conversão alimentar foram observados a 28,6 e 28ºC, respectivamente; b) a temperatura interage com fotoperíodo sobre o desempenho das rãs e seu desenvolvimento gonadal; c) a 27,7ºC (temperatura de conforto térmico) haverá menos rãs dentro d'água; d) a maior temperatura cloacal de rã-touro, 32,1ºC no seco e 33,8ºC dentro d'água, a 35ºC, evidenciou que as rãs se termorregulam; e) os níveis de tetraiodotironina (T4) no plasma decrescem na temperatura de conforto térmico; f) rã-manteiga condiciona-se ao manejo de rotina, reunindo-se ao redor do cocho na hora da alimentação.

Unpalatability of Hyla semilineata tadpoles (Anura) to captive and free-ranging vertebrate predators

This study examines the unpalatability of Hyla semilineata tadpoles, relating this possible defence mechanism to their black, presumably aposematic, colouration. Bullfrog tadpoles (Rana catesbeiana), similar in size to the H. semilineata larvae, were used as controls in the experiments. The palatability of H. semilineata tadpoles was tested by offering the tadpoles to Fish (Hoplias malabaricus), free-ranging passerine birds (Pitangus sulphuratus) and hawks (Buteo magnirostris), and domestic chickens (Gallus domesticus). All predators showed a significant preference towards the control R. catesbeiana tadpoles. However, in experiments with fish, this preference was not significant for tadpole capture, only for their ingestion, suggesting that the fish could not distinguish between the two species before tasting them. Although great kiskadees (P. sulphuratus) preferred the control R. catesbeiana tadpoles, they promptly ingested more than half of the test H. semilineata tadpoles when these were offered alone. The chickens, used as naive predators, clearly learned to avoid the black H. semilineata tadpoles after a few trials. The conspicuous colouration and unpalatability of H. semilineata tadpoles may benefit the individual as well as the group, depending on the predator involved.

Schooling and swimming behaviors of Hyla semilineata tadpoles (Anura, Hylidae)

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

Caracterização de efluentes de viveiros de engorda de rã-touro (lithobates catesbeianus, Shaw, 1802)

Aim: Current analysis characterizes the effluent from bullfrog-rearing ponds during the grow-out phase; Methods: Temperature, pH, dissolved oxygen, electric conductivity, turbidity, total phosphorus, N-NH3, N-NO3, BOD5 and COD and the number of thermotolerant coliforms (Escherichia coli) of the inlet and outlet water of the ponds were analyzed twice a week. Assay consisted of a completely randomized experimental design with two treatments (inlet and outlet water) and six repetitions in a split-plot, coupled to collection over time as subplot; Results: All variables were significantly different (p < 0.05) between treatments and over time (p < 0.05). Average rates of temperature, pH and dissolved oxygen levels of the supply water were higher when compared to those of the effluent. The other variables such as conductivity, turbidity, total phosphorus, ammonia, nitrate, biological oxygen demand, chemical oxygen demand and E. coli were higher in the effluent when compared to rates in the supply water; Conclusions: The management during grow-out phase caused the deterioration of the water quality, with increasing levels of dissolved nutrients and the number of thermotolerant coliform. Ammonia and phosphorus levels in the effluent, caused by waste food, skin and feces, accelerate the eutrophication process of the receiving water body. Further studies on effluent treatment are required.

Primordial germ cells (spermatogonial stem cells) of bullfrogs express sex hormone-binding globulin and steroid receptors during seasonal spermatogenesis

In vertebrate species, testosterone seems to inhibit spermatogonial differentiation and proliferation. However, this androgen can also be converted, via aromatase, into estrogen which stimulates spermatogonial differentiation and mitotic activity. During seasonal spermatogenesis of adult bullfrogs Lithobates catesbeianus, primordial germ cells (PGCs) show enhanced testosterone cytoplasm immunoexpression in winter; however, in summer, weak or no testosterone immunolabelling was observed. The aim of this study was to confirm if PGCs express stem cell markers-alkaline phosphatase (AP) activity and GFRα1 (glial-cell-line-derived neurotrophic factor)-and verify whether testosterone is maintained in these cells by androgen receptors (ARs) and/or sex hormone-binding globulin (SHBG) in winter. Furthermore, regarding the possibility that testosterone is converted into estrogen by PGCs in summer, the immunoexpression of estrogen receptor (ER)β was investigated. Bullfrog testes were collected in winter and in summer and were embedded in glycol methacrylate for morphological analyses or in paraffin for the histochemical detection of AP activity. GFRα1, AR, SHBG and ERβ expression were detected by Western blot and immunohistochemical analyses. The expression of AP activity and GFRα1 in the PGCs suggest that these cells are spermatogonial stem cells. In winter, the cytoplasmic immunoexpression of ARs and SHBG in the PGCs indicates that testosterone is maintained by these proteins in these cells. The cytoplasmic immunoexpression of ERβ, in summer, also points to an ER-mediated action of estrogen in PGCs. The results indicate a participation of testosterone and estrogen in the control of the primordial spermatogonia during the seasonal spermatogenesis of L. catesbeianus. © 2012 S. Karger AG, Basel.

Immunoexpression of aromatase and estrogen receptors β in stem spermatogonia of bullfrogs indicates a role of estrogen in the seasonal spermatogonial mitotic activity

Bullfrog stem spermatogonia, also named primordial germ cells (PGCs), show strong testosterone immunolabeling in winter, but no or weak testosterone immunoexpression in summer. Thus, the role of testosterone in these cells needs to be clarified. In this study, we proposed to evaluate whether PGCs express aromatase and estrogen receptors, and verify a possible role of estrogen in PGCs seasonal proliferation. Testes of male adult bullfrogs, collected in winter (WG) and summer (SG), were fixed and embedded in historesin, for quantitative analysis, or paraffin for immunohistochemistry (IHC). The number of haematoxylin/eosin stained PGCs/lobular area was obtained. Proliferating cell nuclear antigen (PCNA), aromatase, estrogen receptor β (ERβ) and PCNA/ERβ double immunolabeling were detected by IHC. The number of PCNA-positive PGCs and the histological score (HSCORE) of aromatase and ERβ immunolabeled PGCs were obtained. Although the number of PGCs increased significantly in WG, a high number of PCNA-positive PGCs was observed in summer. Moreover, aromatase and ERβ HSCORE was higher in SG than WG. The results indicate that PGCs express a seasonal proliferative activity; the low mitotic activity in winter is related to the maximal limit of germ cells which can be supported in the large lobules. In SG, the increased ERβ and aromatase HSCORE suggests that testosterone is converted into estrogen from winter to summer. Moreover, the parallelism between the high PGCs mitotic activity and ERβ immunoexpression suggest a participation of estrogen in the control of the PGCs seasonal proliferative activity which guarantee the formation of new germ cysts from summer to next autumn. © 2012 Elsevier Inc.

Role of midbrain in the control of breathing in anuran amphibians

The present study was designed to explore systematically the midbrain of unanesthetized, decerebrate anuran amphibians (bullfrogs), using chemical and electrical stimulation and midbrain transections to identify sites capable of exciting and inhibiting breathing. Ventilation was measured as fictive motor output from the mandibular branch of the trigeminal nerve and the laryngeal branch of the vagus nerve. The results of our transection studies suggest that, under resting conditions, the net effect of inputs from sites within the rostral half of the midbrain is to increase fictive breathing frequency, whereas inputs from sites within the caudal half of the midbrain have no net effect on fictive breathing frequency but appear to act on the medullary central rhythm generator to produce episodic breathing. The results of our stimulation experiments indicate that the principal sites in the midbrain that are capable of exciting or inhibiting the fictive frequency of lung ventilation, and potentially clustering breaths into episodes, appear to be those primarily involved in visual and auditory integration, motor functions, and attentional state.