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

The American bullfrog (Rana catesbeiana), recently named Lithobates catesbeianus is currently farmed for commercial purposes throughout various regions of Brazil. Stressful situations such as problems of management, inadequate facilities and environmental changes with consequent reduction of immunity are common in intensive production. The assessments of these situations of stress allow us detect these problems decreasing the injuries causes by confinement. The main objective of this study was to use the biological markers of plasma cortisol and glucose level and hematological parameters to evaluate the response of bullfrog tadpoles submitted to stressed mechanisms of capture and hypoxia. The animals were subjected to three treatments: stress due to individual capture with a hand net; stress due to batch capture with a hand net; and stress due to capture by emptying. The results obtained demonstrated that there were no statistically significant differences in the parameters tested when comparing the treatments with and without exposure to air (normoxia and hypoxia). Based on these results we can conclude that the stressful stimuli tested were not adequate to alter the biomarker tested. For the cortisol, probably this should have occurred due to the synergistic action between this hormone and thyroxin, which induces metamorphosis in these animals.

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

The aim of this study was to evaluate alterations to the physiological profile (cortisol, glycaemia, and blood parameters) of Lithobates catesbeianus caused by the stressors density and hypoxia. The organisms were in the prometamorphosis stage and exposed to different tadpole densities: 1 tadpole/L (T1), 5 tadpoles/L (T2), and 10 tadpoles/L (T3) for 12 days. The blood was collected through the rupture of the caudal blood vessel and collected under normoxia (immediate collection) and hypoxia (after 15 minutes of air exposure) conditions. Cortisol levels rose on the fourth and eighth days of treatment and returned to basal levels by the end of the experiment. The stressor mechanisms tested did not affect glycaemia. White blood cells (total number of lymphocytes, neutrophils, and eosinophils) showed a significant difference at the twelfth day of the experiment when compared with the start of the experiment. We concluded that, under controlled conditions, a density of up to 10 tadpoles/L and air exposure for 15 minutes did not cause harmful physiological alterations during the experimental period. The answer to these stressors maybe was in another hormonal level (corticosterone).

Is hypoxia a stressor to American bullfrog tadpoles?

The aim of this study was to evaluate alterations to the physiological profile of cortisol in pro-metamorphose phase tadpoles of Lithobates catesbeianus exposed to hypoxia stressor in a capture experiment and in a crowding experiment. The capture study was performed by the treatments: stress due to individual capture with a hand net, stress due to batch capture with a hand net and stress due to capture by emptying. Three simultaneous replicates was done witch 12 animals were sampled (6 normoxia - immediately blood collection) and 6 hypoxia - blood collection after 15 min of air exposition) in two collection times with 5 days by intervals. The crowding study was performed by the treatments 1 tadpole L-1, 5 tadpoles L-1 and 10 tadpoles L-1. Three simultaneous replicates was done witch 8 animals (4 normoxia and 4 hypoxia) were sampled in the zero moment (ZM) - blood collection before the experiment, 6 animals/treatment (3 normoxia and 3 hypoxia) to 4 and 8 days and 18 animals/treatment (9 normoxia and 9 hypoxia) to 12 days. The average values to plasmatic cortisol varying from 1.7 to 5.1ng mL-1 (capture study) and 1.0 to 4.2ng mL-1 (crowding study). It concludes that the biomarker tested (cortisol) showed no alterations front of the stressor used. Alternatively, a larger response pattern to these stimuli may have been expressed in another level of an unmeasured hormone (corticosterone). And the bullfrog has great ability to adapt to different management compared to other aquatic organisms, which demonstrates the plasticity of these animals.

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

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

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

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

Predicting the potential distribution of the alien invasive American bullfrog (Lithobates catesbeianus) in Brazil

The non-native invasive anuran Lithobates catesbeianus is presently distributed in Brazil, especially in the Atlantic Rainforest biodiversity hotspot. Here, we use a maximum entropy ecological niche modeling algorithm (i) to model the North American native geographic distribution of this species and (ii) to project that model onto the whole of Brazil. After applying a threshold value that balances commission and omission errors, the projection results suggested high probabilities of occurrence mostly in southern and southeastern Brazil. We also present the first report on the species known distribution in Brazil, showing good agreement with model predictions. If the predictive map is interpreted as depicting invasiveness potential of L. catesbeianus, strategies to prevent further invasion in Brazil should be focused especially in the Atlantic Rainforest biodiversity hotspot.

Is hypoxia a stressor to American bullfrog tadpoles?

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

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

A rã-touro americana (Rana catesbeiana) recentemente denominada Lithobates catesbeianus é cria da com propósito comercial em várias regiões do Brasil. Situações estressantes tais como problemas de manejo, criação inadequada e alterações ambientais com consequente redução da imunidade são comuns em produções intensivas. A avaliação destas situações de estresse permite-nos detectar estes probemas e diminuir as injurias causadas pelo confinamento. O principal objetivo deste estudo foi utilizar os marcadores biológicos de cortisol, glicemia e dados hematológicos para avaliar a resposta de girinos de rã-touro submetidos aos mecanismos estressores de captura e hipóxia. Os animais foram distribuídosem três tratamentos: estresse por captura individual com puçá; estresse por captura em massa com puçá e estresse por captura por escoamento. Os resultados obtidos demostraram não haver diferenças estatisticamente significativas entre os parametros avaliados quando comparou-se os grupos com e sem exposição ao ar (normoxia e hipoxia). Com base nestes resultados pode-se concluir que os estímulos estressores avaliados não foram adequados para alterar os valores plasmáticos dos marcadores biológicos testados. Para o cortisol, isto ocorreu provavelmente em virtude da ação sinérgica deste hormônio e a tiroxina, que induz a metamorfose nestes animais.

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).

Biological invasions and the acoustic niche: the effect of bullfrog calls on the acoustic signals of white-banded tree frogs

Invasive species are known to affect native species in a variety of ways, but the effect of acoustic invaders has not been examined previously. We simulated an invasion of the acoustic niche by exposing calling native male white-banded tree frogs (Hypsiboas albomarginatus) to recorded invasive American bullfrog (Lithobates catesbeianus) calls. In response, tree frogs immediately shifted calls to significantly higher frequencies. In the post-stimulus period, they continued to use higher frequencies while also decreasing signal duration. Acoustic signals are the primary basis of mate selection in many anurans, suggesting that such changes could negatively affect the reproductive success of native species. The effects of bullfrog vocalizations on acoustic communities are expected to be especially severe due to their broad frequency band, which masks the calls of multiple species simultaneously.

Knowing the past to predict the future: land-use change and the distribution of invasive bullfrogs

Biological invasions and land-use changes are two major causes of the global modifications of biodiversity. Habitat suitability models are the tools of choice to predict potential distributions of invasive species. Although land-use is a key driver of alien species invasions, it is often assumed that land-use is constant in time. Here we combine historical and present day information, to evaluate whether land-use changes could explain the dynamic of invasion of the American bullfrog Rana catesbeiana (=Lithobathes catesbeianus) in Northern Italy, from the 1950s to present-day. We used maxent to build habitat suitability models, on the basis of past (1960s, 1980s) and present-day data on land-uses and species distribution. For example, we used models built using the 1960s data to predict distribution in the 1980s, and so on. Furthermore, we used land-use scenarios to project suitability in the future. Habitat suitability models predicted well the spread of bullfrogs in the subsequent temporal step. Models considering land-use changes predicted invasion dynamics better than models assuming constant land-use over the last 50 years. Scenarios of future land-use suggest that suitability will remain similar in the next years. Habitat suitability models can help to understand and predict the dynamics of invasions; however, land-use is not constant in time: land-use modifications can strongly affect invasions; furthermore, both land management and the suitability of a given land-use class may vary in time. An integration of land-use changes in studies of biological invasions can help to improve management strategies.

Predator Survival Tactics and Use of Habitat Cover in Rana Catesbeiana.

Predator-prey relationships are an important aspect of the natural world, and, because of its relevance to survival and natural selection, is an interesting relationship to study. In amphibian larvae, level of activity and landscape use are often what determines the survival as prey. I studied the anti-predator behavior of the North American bullfrog (Rana catesbeiana) tadpoles when presented with dragonfly (Aeshna) larvae, a known predator of tadpoles. Tadpoles were acclimated to four different habitats with varying degrees of habitat cover, and were transferred to a new habitat with a degree of cover equal to one of the acclimation tanks. A restrained predator, and thus its chemical cue, was introduced, and the behavior, particularly the use of the habitat cover to hide from the perceived risk of predation was observed. A significantly higher frequency of inactivity was found in tank I than in II and III, and inactivity followed a general trend of decreasing with increasing habitat cover. Difference in tank cover was not found to have a significant effect on swimming behavior, but did have a significant effect on hiding behavior, which increased with higher availability. Foraging decreased significantly with the addition of a predator, but did not vary significantly with different levels of cover. Hiding behavior and reducing conspicuous behaviors (like foraging) are probably the behaviors that afford the tadpole the most success at eluding a predator in their natural environment.

Mucopolysaccharidosis Type Ii: Identification Of 30 Novel Mutations Among Latin American Patients.

In this study, 103 unrelated South-American patients with mucopolysaccharidosis type II (MPS II) were investigated aiming at the identification of iduronate-2-sulfatase (IDS) disease causing mutations and the possibility of some insights on the genotype-phenotype correlation The strategy used for genotyping involved the identification of the previously reported inversion/disruption of the IDS gene by PCR and screening for other mutations by PCR/SSCP. The exons with altered mobility on SSCP were sequenced, as well as all the exons of patients with no SSCP alteration. By using this strategy, we were able to find the pathogenic mutation in all patients. Alterations such as inversion/disruption and partial/total deletions of the IDS gene were found in 20/103 (19%) patients. Small insertions/deletions/indels (<22 bp) and point mutations were identified in 83/103 (88%) patients, including 30 novel mutations; except for a higher frequency of small duplications in relation to small deletions, the frequencies of major and minor alterations found in our sample are in accordance with those described in the literature.

Compartment Syndrome After South American Rattlesnake (crotalus Durissus Terrificus) Envenomation.

In order to report the outcome of a patient who developed compartment syndrome after South American rattlesnake (Crotalus durissus terrificus) envenomation, confirmed by subfascial pressure measurement and magnetic resonance imaging (MRI). A 63-year-old male was admitted 1 h after being bitten on the right elbow by a large snake, which was not brought for identification. Physical and laboratory features upon admission revealed two fang marks, local tense swelling, paresthesia, intense local pain, hypertension, coagulopathy, and CK = 1530 U/L (RV < 170 U/L). The case was initially treated with bothropic antivenom (80 mL, intravenously), with no improvement. Evolution within 13-14 h post-bite revealed generalized myalgia, muscle weakness, palpebral ptosis, and severe rhabdomyolysis (CK = 126,160 U/L) compatible with envenoming by C. d. terrificus. The patient was then treated with crotalic antivenom (200 mL, intravenously), fluid replacement, and urine alkalinization. Twenty-four-hour post-bite MRI showed marked muscular edema in the anterior compartment of the right forearm, with a high subfascial pressure (40 mmHg) being detected 1 h later. ELISA of a blood sample obtained upon admission, before antivenom infusion, revealed a high serum concentration of C. d. terrificus venom. No fasciotomy was performed and the patient was discharged seven days later without sequelae. Snakebite by C. d. terrificus with subfascial venom injection may lead to increased intracompartmental pressure.

Variability Modeling Of Rainfall, Deforestation, And Incidence Of American Tegumentary Leishmaniasis In Orán, Argentina, 1985-2007.

American tegumentary leishmaniasis (ATL) is a disease transmitted to humans by the female sandflies of the genus Lutzomyia. Several factors are involved in the disease transmission cycle. In this work only rainfall and deforestation were considered to assess the variability in the incidence of ATL. In order to reach this goal, monthly recorded data of the incidence of ATL in Orán, Salta, Argentina, were used, in the period 1985-2007. The square root of the relative incidence of ATL and the corresponding variance were formulated as time series, and these data were smoothed by moving averages of 12 and 24 months, respectively. The same procedure was applied to the rainfall data. Typical months, which are April, August, and December, were found and allowed us to describe the dynamical behavior of ATL outbreaks. These results were tested at 95% confidence level. We concluded that the variability of rainfall would not be enough to justify the epidemic outbreaks of ATL in the period 1997-2000, but it consistently explains the situation observed in the years 2002 and 2004. Deforestation activities occurred in this region could explain epidemic peaks observed in both years and also during the entire time of observation except in 2005-2007.