Orexin-A inhibits fictive air breathing responses to respiratory stimuli in the bullfrog tadpole (Lithobates catesbeianus)

ABSTRACT In pre-metamorphic tadpoles, the neural network generating lung ventilation is present but actively inhibited; the mechanisms leading to the onset of air breathing are not well understood. Orexin (ORX) is a hypothalamic neuropeptide that regulates several homeostatic functions, including breathing. While ORX has limited effects on breathing at rest, it potentiates reflexive responses to respiratory stimuli mainly via ORX receptor 1 (OX1R). Here, we tested the hypothesis that OX1Rs facilitate the expression of the motor command associated with air breathing in pre-metamorphic bullfrog tadpoles (Lithobates catesbeianus). To do so, we used an isolated diencephalic brainstem preparation to determine the contributions of OX1Rs to respiratory motor output during baseline breathing, hypercapnia and hypoxia. A selective OX1R antagonist (SB-334867; 5–25 µmol l−1) or agonist (ORX-A; 200 nmol l−1 to 1 µmol l−1) was added to the superfusion media. Experiments were performed under basal conditions (media equilibrated with 98.2% O2 and 1.8% CO2), hypercapnia (5% CO2) or hypoxia (5–7% O2). Under resting conditions gill, but not lung, motor output was enhanced by the OX1R antagonist and ORX-A. Hypercapnia alone did not stimulate respiratory motor output, but its combination with SB-334867 increased lung burst frequency and amplitude, lung burst episodes, and the number of bursts per episode. Hypoxia alone increased lung burst frequency and its combination with SB-334867 enhanced this effect. Inactivation of OX1Rs during hypoxia also increased gill burst amplitude, but not frequency. In contrast with our initial hypothesis, we conclude that ORX neurons provide inhibitory modulation of the CO2 and O2 chemoreflexes in pre-metamorphic tadpoles.

Use of a native predator for the control of an invasive amphibian

Context The control of invasive alien species is essential for securing native biodiversity. As for the American bullfrog, Lithobates catesbeianus (Shaw 1802), suspected to cause ecological damage around large parts of the globe, comprehensive management techniques are currently absent. Aims To fill this gap, opportunities arising from biomanipulation of permanent water bodies inhabited by fish were explored. Methods A multi-annual experiment was performed in small and shallow ponds, and effects of complete drawdown (with amphibian and fish removal) and predation (introduction of originally occurring native northern pike, Esox lucius) on non-indigenous bullfrogs were investigated. Key results The presence of pike lead to a strong decline in bullfrog tadpole numbers, whereas no effect of drawdown was observed. Also, communities receiving pike harboured substantially less small and mostly planktivorous fish species (e.g. pumpkinseed, Lepomis gibbosus, and topmouth gudgeon, Pseudorasbora parva). Conclusions The reduction in bullfrog tadpoles may be assigned to both direct and indirect effects induced by pike. First, direct pike predation on tadpoles was observed. Second, as the occurrence of macroinvertebrate-feeding pumpkinseed was low in the presence of pike, the indirect effect of predation by macroinvertebrates on tadpoles may significantly increase, leading to tadpole decline. Implications Biomanipulation of permanent water bodies inhabited by fish can thus be regarded as a candidate for effective and sustainable control of invasive bullfrog. Piscivorous fish introduction may be applied in the specific type of water body, but requires careful consideration of the indigenous status of the introduced species, angling purposes, or specific nature values.