The role of hind limb flexor muscles during swimming in the toad, Bufo marinus

Most work examining muscle function during anuran locomotion has focused largely on the roles of major hind limb extensors during jumping and swimming. Nevertheless, the recovery phase of anuran locomotion likely plays a critical role in locomotor performance, especially in the aquatic environment, where flexing limbs can increase drag on the swimming animal. In this study, I use kinematic and electromyographic analyses to explore the roles of four anatomical flexor muscles in the hind limb of Bufo marinus during swimming: m. iliacus externus, a hip flexor; mm. iliofibularis and semitendinosus, knee flexors; and m. tibialis anticus longus, an ankle flexor. Two general questions are addressed: (1) What role, if any, do these flexors play during limb extension? and (2) How do limb flexors control limb flexion? Musculus iliacus externus exhibits a large burst of EMG activity early in limb extension and shows low levels of activity during recovery. Both m. iliofibularis and m. semitendinosus are biphasically active, with relatively short but intense bursts during limb extension followed by longer and typically weaker secondary bursts during recovery. Musculus tibialis anticus longus becomes active mid way through recovery and remains active through the start of extension in the next stroke. In conclusion, flexors at all three joints exhibit some activity during limb extension, indicating that they play a role in mediating limb movements during propulsion. Further, recovery is controlled by a complex pattern of flexor activation timing, but muscle intensities are generally lower, suggesting relatively low force requirements during this phase of swimming.     

Leptospires in the marine toad (Bufo marinus) on Barbados

Leptospires were isolated from the kidneys of four of 211 toads (Bufo marinus) caught on Barbados. Two of the isolates were identified as Leptospira interrogans serovar bim in the Autumnalis serogroup (the most common cause of leptospiral illness on Barbados), and two as possibly new serovars in the Australis serogroup. Sera from 198 of the toads were examined by the leptospire microscopic agglutination test. Forty-two (21%) were positive at titers of greater than or equal to 1:100, and 54 (27%) at greater than or equal to 1:50. The predominating serogroups were Australis (50%), Autumnalis (23%) and Panama (13%). The agglutination tests on the culture-positive toads showed that serologic studies alone may be of limited value in these animals. Bufo marinus can harbor pathogenic leptospires, and it may be a significant source of the Autumnalis serogroup infections in the Caribbean.     

Microbodies (peroxisomes) in the toad,Bufo marinus

Summary Microbodies (peroxisomes), a group of cytoplasmic organelles enriched in catalase, are demonstrated in the toad, Bufo marinus, by light and electron microscopy by means of a cytochemical staining procedure that demonstrates the peroxidatic activity of catalase with diaminobenzidine (DAB). Amphibian microbodies are similar to those of other classes in their fine structure and localization in hepatocytes and kidney, where they are prominent in the proximal tubular cells. Nucleoids are present only in renal microbodies. In the proximal renal tubule an unusual group of large brown granules are identified as lysosomes by their acid phosphatase, -glucosaminidase and -glucuronidase activities.This work was supported by U.S. Public Health Service Grants Nos. NS-06856 and HD 00674. We wish to thank Dr. Richard M. Hays who generously supplied us with toads; Dr. Alex B. Novikoff for making available facilities for ultramicrotomy, Miss Betty De Prest for technical assistance; Miss Marianne Van Hooren for preparation of the photomicrographs.     

Osmoregulation of the cane toad, Bufo marinus, in salt water

Adult cane toads, B. marinus, survived in salinities up to 40% sea-water (SW). Pre-exposure to 30, then 40% SW, increased the survival time of toads in 50% SW. Plasma from toads acclimated to salt water is hyperosmotic to the environment--a result of increased plasma sodium, chloride and urea concentrations. When toads were placed in tap-water and 20% SW, all significant changes to plasma sodium, chloride, urea and osmotic pressure occurred within the first 2 days of exposure. When toads were placed in 30 and 40% SW environments, the increases in plasma sodium and chloride concentrations occurred within the first 2 days of exposure while urea and total osmotic pressure continued to rise until some time between 2 and 7 days exposure.     

Parallel metabotropic pathways in the heart of the toad, Bufo marinus

This study examined the transduction pathways activated by epinephrine in the pacemaker region of the toad heart. Recordings of membrane potential, force, and intracellular Ca(2+) concentration ([Ca(2+)](i)) were made from arrested toad sinus venosus. Sympathetic nerve stimulation activated non-alpha-, non-beta-adrenoceptors to evoke a membrane depolarization and a transient increase in [Ca(2+)](i). In contrast, the beta-adrenoceptor agonist isoprenaline (10 microM) caused membrane hyperpolarization and decreased [Ca(2+)](i). The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (0.5 mM) mimicked the isoprenaline-evoked membrane hyperpolarization. Epinephrine (10-50 microM) caused an initial membrane depolarization and an increase in [Ca(2+)](i) followed by membrane hyperpolarization and decreased [Ca(2+)](i). The membrane depolarizations evoked by sympathetic nerve stimulation or epinephrine were abolished either by the phospholipase C inhibitor U-73122 (20 microM) or by the blocker of D-myo-inositol 1,4,5,-trisphosphate-induced Ca(2+) release, 2-aminoethoxydiphenyl borate (2-APB, 60 microM). Neither U-73122 nor 2-APB had an affect on the membrane hyperpolarization evoked by beta-adrenoceptor activation. These results suggest that in the toad sinus venosus, two distinct transduction pathways can be activated by epinephrine to cause an increase in heart rate.     

Peptide-containing nerves in the urinary bladder of the toad,Bufo marinus

The distributions of peptide-containing nerves in the urinary bladder of the toad, Bufo marinus, were studied by means of fluorescence immunohistochemistry of whole-mount preparations. The bundles of smooth muscle in the bladder are well supplied by varicose nerve fibres displaying somatostatin-like immunoreactivity; these fibres probably arise from intrinsic perikarya. The urinary bladder also has a well-developed plexus of nerves containing substance P-like immunoreactive material; these elements probably represent sensory nerves of extrinsic origin. Nerve fibres showing immunoreactivity to vasoactive intestinal polypeptide (VIP) or enkephalin are rare within the urinary bladder of the toad. It is considered unlikely that any of these peptides directly mediates the hyoscine-resistant excitatory response of the smooth muscle to nerve stimulation in the toad bladder.     

Inward rectification in muscle fibres of the toad Bufo marinus

Inward rectification of the resting potassium conductance was studied in skeletal muscle fibres of the toad Bufo marinus. This conductance was shown to be blocked by Ba and Cs and located both in the surface membrane and the membranes of the tubular system. Some differences were found between the properties of this conductance channel in Bufo marinus and those reported for various species of Rana. The possible adaptative value of these differences is pointed out.     

Active phosphate transport across the urinary bladder of the toad, Bufo marinus.

Net transport of inorganic phosphate occurs in the absence of an electrochemical gradient from the mucosal to the serosal bathing solution in the isolated toad urinary bladder. This transport can be inhibited by metabolic inhibitors. The magnitude of this transport can be altered by changes in phosphate concentration or by the addition of parathyroid hormone.     

Fine structure of the photosensitive iris of the toad, Bufo marinus

The iris of the toad Bufo marinus is directly photosensitive and will constrict in response to light striking only the iris. This is true even when the iris is isolated from the rest of the eye, and therefore from reflex neuronal influences initiated in the retina. This autonomous response is probably mediated by the sphincter pupillae muscle, since no specialized photoreceptors are present in the iris, nor does the sphincter exhibit any specializations likely to subserve a purely photoreceptive function. The photosensitive sphincter appears typical of smooth muscle and, like mammalian sphincters, possesses many intercellular junctions. The iris possesses a well-developed neuronal plexus with fibers projecting into the sphincter muscle layer. Nerve terminals contain small, agranular (30-70nm) and large, dense-cored (80-120nm) vesicles. No consistent postsynaptic specializations are seen on any cells of the iris, including the cells of the sphincter muscle. The anterior pigment epithelial cells of the iris appear specialized and resemble the myoepithelial dilator muscle described by Kelly and Arnold ('72) for the iris of rats.     

Natriuretic peptide receptors in the central vasculature of the toad, Bufo marinus

Natriuretic peptide receptors in the central vasculature of the toad, Bufo marinus, were characterized using autoradiographical, molecular, and physiological techniques. Specific 125I-rat ANP binding sites were present in the carotid and pulmonary arteries, the lateral aorta, the pre- and post-cava, and the jugular vein, and generally occurred in each layer of the blood vessel. The 125I-rat ANP binding was partially displaced by the specific natriuretic peptide receptor C ligand, C-ANF, which indicates the presence of two types of natriuretic peptide receptors in the blood vessels. This was confirmed by a RT-PCR study, which demonstrated that guanylyl cyclase receptor (NPR-GC) and NPR-C mRNAs are expressed in arteries and veins. An in vitro guanylyl cyclase assay showed that frog ANP stimulated the production of cGMP in arterial membrane fractions. Physiological recordings from isolated segments of the carotid and pulmonary arteries and the lateral aorta, which had been pre-constricted with arginine vasotocin, showed that rat ANP, frog ANP and porcine CNP relaxed the vascular smooth muscle with relatively similar potency. Together, the data show that the central vasculature contains two types of natriuretic peptide receptors (NPR-C and NPR-GC) and that the vasculature is a target for ANP and CNP.     

Plasma cathepsin activity and the role of hypovolemic shock in dehydrational death in the toad, Bufo marinus

1. The pH- and activator-dependence of toad plasma cathepsin activity assayed by hemoglobin digestion was characteristic of cathepsins B1 and D. 2. Dehydration, even to the point of death, did not produce a significant elevation of plasma cathepsin activity over controls. 3. Toads were remarkably resistant to the effects of splanchnic artery ligation, which also did not produce significantly higher plasma cathepsin levels. 4. Cardiac depression via the production of a myocardial depressant factor by cathepsins does not appear to be an important factor in dehydrational death in toads.     

Renal vascular anatomy of the toad (Bufo marinus)

The renal vasculature of the toad, Bufo marinus, was studied mainly by means of scanning electron microscopy of vascular corrosion casts. All arterial branches terminated in a glomerulus. Each glomerulus was supplied by only one afferent arteriole. No shunts between afferent and efferent arterioles were observed. The glomerular channels appeared to be permanent capillaries. No evidence supporting the theory of freely shifting glomerular blood channels was found. Efferent arterioles radiated out towards the dorsal surface of the kidney where they connected with peritubular vessels. The renal portal veins produced an anastomosing plexus on the dorsal surface of the kidney, giving rise to the peritubular vessels. Peritubular vessels ran radially toward the ventral surface of the kidney, where they formed the roots of the renal veins. Attention is drawn to the possibility of hairpin countercurrent exchange between the capillary-like efferent arterioles and the peritubular vessels in the dorsal kidney.     

Adrenalin and hydroxytyramine in the parotid gland venom of the toad,Bufo marinus

Preparations from the parotid gland venom of the toad Bufo marinus have been analyzed by paper chromatography for the presence of adrenalin precursors. In addition to adrenalin itself, hydroxytyramine was identified and a second compound which appeared to be an adrenalin ester—perhaps lactyl-adrenalin. Two other compounds which appeared on the chromatogram could not be identified. Neither noradrenalin nor another possible precursor, epinine, appeared to be present.     

Behavioral thermoregulation of the toad, Bufo marinus: effects of air humidity

Two experiments were performed. The first tested the hypothesis that the toad, Bufo marinus, will select a lower ambient temperature under dry environmental conditions. This behavioral response would reduce evaporative water loss and facilitate survival on land. The second experiment measured the effects of temperature on evaporative water loss. In the first experiment, toads were placed in a thermal gradient (11-40 degrees C) for 3 days. On days 1 and 3, water-filled dishes were placed along the temperature gradient and humid air was circulated through the chamber. On day 2, water dishes were removed, and dry air was circulated through the chamber. Body temperature (Tb) was recorded with a cloacal thermistor. Selected Tb was approximately 8.6 degrees C lower during the dry conditions than during the humid conditions. The behavioral hypothermia took about 6 h to develop. In the second experiment, a reduction in Tb from 17.7 to 12 degrees C reduced evaporative water loss by 42%. Consequently, behavioral hypothermia of the toad is an important adaptation to dry environmental conditions.     

Active phosphate transport across the urinary bladder of the toad,Bufo marinus

Summary Net transport of inorganic phosphate occurs in the absence of an electrochemical gradient from the mucosal to the serosal bathing solution in the isolated toad urinary bladder. This transport can be inhibited by metabolic inhibitors. The magnitude of this transport can be altered by changes in phosphate concentration or by the addition of parathyroid hormone.This work was presented in part at the Fall Meeting of the American Physiological Society Proceedings, October, 1975; Abstract inThe Physiologist, 1975,18:384.     

Weber and noise adaptation in the retina of the toad Bufo marinus

Responses to flashes and steps of light were recorded intracellularly from rods and horizontal cells, and extracellularly from ganglion cells, in toad eyecups which were either dark adapted or exposed to various levels of background light. The average background intensities needed to depress the dark-adapted flash sensitivity by half in the three cell types, determined under identical conditions, were 0.9 Rh*s-1 (rods), 0.8 Rh*s-1 (horizontal cells), and 0.17 Rh*s-1 (ganglion cells), where Rh* denotes one isomerization per rod. Thus, there is a range (approximately 0.7 log units) of weak backgrounds where the sensitivity (response amplitude/Rh*) of rods is not significantly affected, but where that of ganglion cells (1/threshold) is substantially reduced, which implies that the gain of the transmission from rods to the ganglion cell output is decreased. In this range, the ganglion cell threshold rises approximately as the square root of background intensity (i.e. in proportion to the quantal noise from the background), while the maintained rate of discharge stays constant. The threshold response of the cell will then signal light deviations (from a mean level) of constant statistical significance. We propose that this type of ganglion cell desensitization under dim backgrounds is due to a post-receptoral gain control driven by quantal fluctuations, and term it noise adaptation in contrast to the Weber adaptation (desensitization proportional to the mean background intensity) of rods, horizontal cells, and ganglion cells at higher background intensities.     

Introduced animals and their parasites: The cane toad,Bufo marinus,in Australia

Abstract The impact of introducing animals into an established ecosystem can be directly observed through predator-prey and competition interactions. The impact of animals via more obscure relationships, such as the host-parasite relationship, are generally not considered. The cane toad Bufo marinus (Linnaeus) was introduced to Australia in 1935. Despite intensive research into many aspects of the biology of the toad, there has been no systematic survey of the parasite fauna of B. marinus in Australia. It is unknown exactly what parasites the toad may have introduced to Australia and also the range of parasites that may have adapted to the toad from native fauna since its introduction. The provisional conclusion from this review is that all the helminth parasites so far recorded from B. marmus in Australia have been acquired from local hosts. The interaction of toads and nativa fauna via their parasites remains unknown.