Diferentes níveis protéicos na dieta de girinos de rã-touro (Lithobates catesbeianus) e seus efeitos sobre o tecido hepático / Different protein levels in the diet of bullfrog tadpoles (Lithobates catesbeianus) and its effects on the liver tissue

Para verificação da ocorrência de alterações morfológicas no fígado, foram coletados dez animais de cada dieta, no 30º dia (1º período experimental) e no 60º dia (2º período experimental), sendo esse, o último dia do período experimental e início do clímax da metamorfose. Os girinos foram distribuídos em delineamento experimental inteiramente casualizado (DIC) com três tratamentos: dieta experimental com 32,68% de proteína bruta e duas dietas comerciais com 37,92% e 57,53% de proteína bruta. Os órgãos coletados foram fixados em solução de Bouin e depois submetidos às práticas da rotina histotecnológica e coradas com hematoxilina - Eosina (HE). Na avaliação microscópica do fígado foram encontradas diversas alterações morfológicas como: Desorganização e vacuolização de hepatócitos, infiltrado inflamatório mononuclear periportal, multifocal e difuso, infiltrado inflamatório eosinofílico, congestão, hemorragia, hemólise e necrose. As alterações hepáticas encontradas neste estudo sugerem que as dietas utilizadas para os girinos de rã-touro, não atenderam suas necessidades dietéticas, afetando a homeostasia dos mesmos, comprometendo assim, sua sanidade.(AU)

To verify the occurrence of liver morphological changes, bullfrog tadpoles were fed three diets: an experimental diet with 32.68% crude protein and two commercial diets with 37.92% and 57.53% crude protein. Tadpoles were distributed into a completely randomized design (CRD) with three treatments and four replications. Liver samples were collected twice, on the 30th and 60th day of the experiment. Tadpole livers were fixed in Bouin's solution, then subjected to routine histotechnology procedures, and stained with hematoxylin - eosin (HE). Microscopic evaluation of liver tissue showed several morphological changes like disorganization and vacuolization of hepatocytes; periportal, multifocal and diffuse mononuclear inflammatory infiltrate, eosinophilic inflammatory infiltration, congestion, hemorrhage, hemolysis and necrosis. The liver changes found in this study suggest that diets used for bullfrog tadpoles, did not meet their dietary needs, what affected their homeostasis, thus compromising their health.(AU)

Effects of the organophosphorus pesticide Folisuper 600 (methyl parathion) on the heart function of bullfrog tadpoles, Lithobates catesbeianus (Shaw, 1802)

Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles performance, jeopardizing their survival.

Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.

Effects of the organophosphorus pesticide Folisuper 600 (methyl parathion) on the heart function of bullfrog tadpoles, Lithobates catesbeianus (Shaw, 1802) / Efeitos do pesticida organofosforado Folisuper 600 (metil paration) sobre a função cardíaca de girinos de rã-touro, Lithobates catesbeianus (Shaw, 1802)

Abstract The aim of this work was to evaluate whether the heart function of bullfrog tadpoles (25 Gosner stage) is affected by their acute exposure (48 h) to a sub-lethal concentration (10 µg.L–1) of the active principle of the organophosphorus pesticide Folisuper 600R (methyl parathion - MP). Our results demonstrated that MP causes not only a reduction in tadpoles’ cardiac ventricular mass, resulting in a marked reduction in their cardiac twitch force, but also impairs their swimming performance, irrespective of increasing their heart rate. Together, these findings indicate that low and realistic concentration of MP have a negative impact on tadpoles’ performance, jeopardizing their survival.

Resumo O objetivo do presente estudo foi avaliar se a função cardíaca de girinos de rãs-touro (estágio 25 de Gosner) é afetada pela exposição aguda (48h) a uma concentração sub-letal (10 µg.L–1) do princípio ativo do pesticida organofosforado Folisuper 600R (metil paration – MP). Nossos resultados demonstraram que o MP ocasionou não apenas uma redução na massa ventricular cardíaca dos girinos, como também provocou uma redução na sua força de contração cardíaca e de seu desempenho natatório, a despeito de ter sido observado um aumento de sua freqüência cardíaca. Conjuntamente, os achados aqui obtidos indicam que uma baixa e realística concentração de MP exerce um impacto negativo sobre o desempenho dos girinos, ameaçando sua sobrevivência.

Is hypoxia a stressor to American bullfrog tadpoles? / A hipóxia é um estressor para girinos de rã-touro?

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.

Avaliou-se o cortisol em girinos de rã-touro (Lithobates catesbeianus), no estágio de pró-metamorfose ao mecanismo estressor de hipóxia juntamente com captura (1º experimento) e adensamento (2º experimento). O experimento de captura foi composto pelos tratamentos: Captura individual com puçá, Captura em massa com puçá e Captura por escoamento, com 3 réplicas simultâneas onde 12 organismos foram amostrados (6 normoxia - coleta imediata de sangue e 6 hipoxia - coleta de sangue após 15 min de exposição ao ar) em 2 tempos de coleta com intervalo de 5 dias. O experimento de densidade foi composto pelos tratamentos: 1 girino L-1, 5 girinos L-1 e 10 girinos L-1, com 3 réplicas simultâneas onde 8 animais (4 normoxia e 4 hipoxia) foram amostrados no momento zero (MZ) - coleta de sangue anterior ao experimento, 6 animais/tratamento (3 normoxia e 3 hipoxia) para 4 e 8 dias e 18 animais/tratamento (9 normoxia e 9 hipoxia) para 12 dias. Os valores médios para o cortisol plasmático foram de 1,7 a 5,1ng mL-1 (Experimento de Captura); e 1,0 a 4,2ng mL-1 (Experimento de Densidade). Conclui-se que o marcador biologico de estresse utilizado (cortisol) não foi alterado pelo agente extressor. Alternativamente a resposta a este estímulo pode ser expressa em outro nível hormonal (corticosterona). E a rã-touro apresenta ótima capacidade de se adaptar aos diferentes manejos se comparados a outros organismos aquáticos, o que demonstra a plasticidade destes animais.

Identification of Elizabethkingia meningoseptica from American bullfrog (Rana catesbeiana) farmed in Sabah, Malaysia using PCR method and future management of outbreak

Aims: High demand for frog meat in Malaysia especially the American bullfrog (Rana catesbeiana) has promoted intensive farming of the animal. However, the farming of American bullfrog is restricted by the occurrence of diseases. This study reports the first isolation of Elizabethkingia meningoseptica from specimens of American bullfrog that suffer from cataract and ‘red-leg’ syndrome. Methodology and Result: The pathogen was isolated from eyes and internal organs (liver, kidney and spleen) of the diseased bullfrog specimens. All the bacterial isolates were subjected to phenotypic characterization and antibiotic susceptibility assay, and further identified by using the 16S rDNA sequencing analysis. We designed two pair of specific PCR primers (22-25 mers) which are complimentary to the β-lactamase gene in the reference strain of E. meningoseptica ATCC49470. The result showed all the bacterial isolates shared similar phenotypic characters and antibiotic susceptibility. BLAST analysis of the 16S rDNA sequences indicated that the bacterial isolates had very high sequence homology (100%) with E. meningospetica ATCC49470 and E. meningoseptica isolates from mosquito. The two PCR primers were very specific to E. meningoseptica isolates of this study. Conclusion, significance and impact of study: This is the first isolation and characterization of bacterial pathogen, E. meningoseptica in cultured American bullfrog (Rana catesbeina) that suffered from eye cataract and ‘red-leg’ syndrome in Sabah, Malaysia. It is suspected that one of the possible transmission routes of the bacterial pathogen could be via mosquito bites. The findings suggest that there is urgent requirement for standard guideline of good farming practice to be adopted in frog farms throughout the country. Such a guideline can help in minimizing economic losses, preventing transmission of the zoonotic bacterial pathogen to farm workers, and sustaining the industry in Malaysia and upgrading frog meat quality for international market.

Un modelo anfibio para el estudio evolutivo del control respiratorio / Anphibian model study for evolutionary respiratory control

Los enormes progresos de la medicina perinatal han permitido la sobrevivencia de recién nacidos prematuros en estadios de desarrollo cada vez más precoces. Estos pequeños pacientes requieren de medios muy sofisticados y costosos para paliar su inmadurez respiratoria. Además de las complicaciones asociadas al desarrollo pulmonar, la inestabilidad respiratoria y las apneas asociadas a la inmadurez de los circuitos nerviosos productores del control respiratorio, contribuyendo éstos últimas, como factor principal de morbilidad, a las prolongadas hospitalizaciones que presenta esta tan vulnerable población. Éstos imperativos médicos combinados a la curiosidad fisiológica han contribuido al desenvolvimiento de novedosas investigaciones en neurobiología respiratoria. Si bien la mayoría de los trabajos realizados en éste campo utilizan a los múridos como modelos experimentales, las investigaciones recientes efectuadas a partir del tronco cerebral aislado de anfibios (Rana catesbeiana) han revelado las ventajas técnicas de este modelo animal, para mostrar los principales fenómenos que rigen el funcionamiento y el desarrollo del sistema de control respiratorio entre los vertebrados. El presente artículo se propone repasar los avances más recientes realizados en la investigación de la neurobiología evolutiva del control respiratorio, prestando una atención particular a la respiración episódica, así como al rol que desempeñan la modulación serotoninérgica y Gabaérgica de la actividad respiratoria en el curso del desarrollo.

Recent perinal advances have made possible that premature newborns survive increasingtly in earlier developmental stages. This babies requires sophisticated and costly critical intensive care to address the problems associated with inmadurity of the respiratory system. In addition respiratory instability and apnea reflecting inmaturity of the respiratory control system are major causes of morbidity and prolonged hospitalization in this highly vulnerable group of patients. These concerns have contributed to the development of research in respiratoy evolutionary neurobiology. While the majority of researchers working in this field use rodents as an animal model, recent research using in vitro brainstem preparations from bullfrog (Rana catesbeiana) have reveled the technical advantages of this model to study the basic principles underlying respiratory control and its ontogeny between vertebrates. The present article review the recent advances in the area of research with special interest on episodic breathing and the role of serotoninergic and GABAergic modulation of respiratory control during development.