The physical act of eating or feeding involves the coordinated action of several organs like eyes and jaws, and associated neural networks. Moreover, the activity of the neural networks controlling jaw movements (branchiomotor circuits) is regulated by the visual, olfactory, gustatory and hypothalamic systems, which are largely well characterized at the physiological level. By contrast, the behavioral output of the branchiomotor circuits, and the functional consequences of disruption of these circuits by abnormal neural development are poorly understood. To begin to address these questions, we sought to evaluate the feeding ability of zebrafish larvae, a direct output of the branchiomotor circuits, and developed a qualitative assay for measuring food intake in zebrafish larvae at 7 days post fertilization. We validated the assay by examining the effects of ablating the branchiomotor neurons. Metronidazole-mediated ablation of nitroreductase-expressing branchiomotor neurons resulted in a predictable reduction in food intake without significantly affecting swimming ability, indicating that the assay is robust. Laser-mediated ablation of trigeminal motor neurons resulted in a significant decrease in food intake, indicating that the assay is sensitive. Importantly, in larvae of a genetic mutant with severe loss of branchiomotor neurons, food intake was abolished. These studies establish a foundation for dissecting the neural circuits driving a motor behavior essential for survival.
The medicinal leech, Hirudo medicinalis, has been recognized by plastic surgeons for its ability to reduce congestion and improve blood flow in grafted skin flaps by withdrawing excess blood. The leech's ability to digest blood is due to the presence of Aeromonas hydrophila in the gut of the leech. In this report we describe the occurrence of Aeromonas hydrophila septicaemia in rabbits used in a study to evaluate the efficacy of the medicinal leech on abdominal skin flap survival. In New Zealand White rabbits, twin 5 × 10 cm abdominal skin flaps were prepared and the epigastric vein was ligated to produce a model of venous congested skin. After 12 hours, a leech was applied to one of the congested abdominal flaps and skin survivability was assessed and compared with the unleeched flap. Five rabbits died acutely approximately 24 hours post-operatively. Gross necropsy, bacteriology and histopathology findings indicated that A. hydrophila was the causative agent of the septicaemia-toxaemia syndrome with an associated pneumonia and typhlitis. Review of the management and the experimental surgery protocol suggested that the two major disposing factors of Aeromonas septicaemia in these rabbits were stress and excessive prophylactic administration of penicillin.
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