Galanin: presence and distribution in the brain and pituitary of Rhinella arenarum (Amphibia: Anura) during development

The immunohistochemical distribution of galanin (Gal) in the brain and pituitary of Rhinella arenarum was studied during development. Gal-immunoreactivity was first observed in the brain just after hatching in anterior preoptic area, infundibular area, median eminence and pars distalis of the pituitary as well as in the olfactory epithelium. At the beginning of prometamorphosis new Gal-immunoreactive (ir) cells were observed in the olfactory nerve and bulb. Later in prometamorphosis new Gal-ir cells were observed in the telencephalon, suprachiasmatic nucleus, rostral rhombencephalon and in the pars nervosa of the pituitary. The most numerous accumulations of Gal-ir neurons throughout the larval development were observed in the ventral hyphothalamus where numerous Gal-ir cells of cerebrospinal fluid-contacting type were found. During metamorphic climax and soon after we did not detect Gal-ir neurons in the pallium, medial or pretectal dorsal thalamus. In the median eminence and pars distalis of the pituitary many Gal-ir fibers were found during development indicating that Gal may play a role in the modulation of hypophyseal secretion. Furthermore, the distribution of Gal-ir elements observed throughout larvae development indicates that galaninergic system maturation continues until sexual maturity.     

Acknowledgments

One of the major causes of worldwide amphibian declines is a skin infection caused by a pathogenic chytrid fungus (Batrachochytrium dendrobatidis). This study documents the interactions between this pathogen and a susceptible amphibian host, the boreal toad (Bufo boreas). The amount of time following exposure until death is influenced by the dosage of infectious zoospores, duration of exposure, and body size of the toad. The significant relation between dosage and the number of days survived (dose-response curve) supports the hypothesis that the degree of infection must reach a particular threshold of about 10⁷–10⁸ zoosporangia before death results. Variation in air temperature between 12°C and 23°C had no significant effect on survival time. The infection can be transmitted from infected to healthy animals by contact with water containing zoospores; no physical contact between animals is required. These results are correlated with observations on the population biology of boreal toads in which mortalities associated with B. dendrobatidis have been identified.     

Polymerase chain reaction primers for microsatellite loci in the Common Toad Bufo bufo

The Common Toad Bufo bufo is a wide‐ranging species, with a distribution encompassing much of Europe and Asia. Few molecular studies have been undertaken on this species, and only one polymorphic microsatellite locus has been identified. Therefore, little is known about the genetic variability within and between B. bufo populations. The value of such information is essential for monitoring the species and its environment. This paper reports the characterization of 15 B. bufo microsatellite loci using individuals from a Spanish and UK sample.     

中华蟾蜍血细胞的观察

应用常规血涂片Wright氏染色、活体观察和血细胞计数法研究中华蟾蜍（Bufo　gargarizans）的血细胞形态。结果表明:红细胞有有核和无核两种类型。有核红细胞较大,椭圆形,平均19.41×14.25μm,细胞核染色质密集。白细胞圆形或椭圆形,直径比红细胞长径短,核比例一般较大。白细胞中,淋巴细胞最多,其次为中性粒细胞和单核细胞,嗜酸性粒细胞少,嗜碱性粒细胞极少。红细胞和淋巴细胞均有直接分裂现象。红细胞平均值为7.67×105/mm3,白细胞平均值为0.75×105/mm3。血栓细胞纺锤形或卵圆形,胞质少或裸核状,推侧为红细胞的衍生物。     

The toad fly Lucilia bufonivora: its evolutionary status and molecular identification

The blow fly genus Lucilia is composed largely of saprophages and facultative myasis agents, including the economically important species Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) and Lucilia sericata (Meigen). Only one species is generally recognized as an obligate agent of myiasis, Lucilia bufonivora Moniez, and this is an obligate parasite of toads. Lucilia silvarum (Meigen), a sister species, behaves mainly as a carrion breeder; however, it has also been reported as a facultative parasite of amphibians. Morphologically, these species are almost identical, and historically this has led to misidentification, taxonomic ambiguity and a paucity of studies of L. bufonivora. In this study, dipterous larvae were analysed from toad myiasis cases from the U.K., The Netherlands and Switzerland, together with adult specimens of fly species implicated in amphibian parasitism: L. bufonivora, L. silvarum and Lucilia elongata Shannon (from North America). Partial sequences of two genes, cox1 and ef1α, were amplified. Seven additional blow fly species were analysed as outgroups. Bayesian inference trees of cox1, ef1α and a combined‐gene dataset were constructed. All larvae isolated from toads were identified as L. bufonivora and no specimens of L. silvarum were implicated in amphibian myiasis. This study confirms L. silvarum and L. bufonivora as distinct sister species and provides unambiguous molecular identification of L. bufonivora.     

Host plant toxins and unpalatability of Neacoryphus bicrucis (Hemiptera: Lygaeidae)

Abstract. 1. Host plant choice of the seed bug, Neacoryphus bicrucis Say (Hemiritera, Lygaeidae), was evaluated in four different Georgia habitats. N. bicrucis utilized only species of Senecio as host plants at a granite outcrop and at old fields on the coastal plain, in the piedmont, and in the Blue Ridge Mountains.
2. In laboratory tests N. bicrucis is distasteful to the green anole lizard, Anolis carolinensis Voight. Only six of ninety field-collected bugs were eaten in palatability trials while eighty-four of the remaining ninety attacked bugs were rejected by lizards without apparent harm. However, the bugs were palatable to Fowler's toads, Bufo woodhousei fowleri Hinckley, which ate all thirty-six bugs offered to them.
3. N. bicrucis selectively sequesters pyrrolizidine alkaloids from Senecio spp. These may cause the insects' distastefulness to green anoles. In palatability trials, lizards ate all twenty insects reared on sunflower seeds, Helianthus annus, but rejected all twenty reared on Senecio smallii.     

NPY- and CGRP-like immunoreactive nerve fibers in the testis and mesorchium of the toad (Bufo arenarum)

The presence and distribution of peptidergic nerve fibers were studied in the testis and mesorchium of the toad by means of immunohistochemistry. Cryostat sections of the testis and whole-mount preparations of mesorchia were immunostained with antisera to calcitonin gene-related peptide (CGRP) and neuropeptide tyrosine (NPY). After leaving the mesorchium CGRP-immunoreactive (IR) fibers were seen predominantly running in between the seminiferous tubules. In addition, a small population of CGRP-IR nerve fibers formed thin plexuses around blood vessels. Conversely, NPY-like immunoreactivity predominated in nerve fibers that formed dense plexuses around vessels both in the mesorchium and testis. Additionally, some single NPY-IR nerve fibers could be seen in both structures studied. The functional significance of these peptidergic systems in the testis of the toad remains to be analyzed.     

The extra-limital spread of an invasive species via 'stowaway' dispersal: toad to nowhere?

The mechanisms by which invasive species spread through new areas can influence the spatial scale of their impact. Although previous research has focused on 'natural' dispersal rates following initial introductions, human-aided inadvertent dispersal by 'stowaways' on commercial and domestic transport is thought to be a major contributor to long-distance dispersal. Few data exist to support this assumption. Cane toads Bufo marinus were introduced to north-eastern Australia in 1935, and have since dispersed rapidly through the tropics. Based on information accumulated by community groups in Sydney, 400 km south of the cane toads' current Australian distribution, we document high rates of translocation (at least 50 toads arriving in Sydney per year). Most toads were translocated on commercial truck transport carrying landscaping and building materials from the current range of the cane toads in New South Wales and Queensland, and resulted in highly clumped locations of toad arrival reflecting primary truck transport destinations. Most introductions involved single toads (68 of 102 translocation events), but some introductions involved two to 19 animals. Adults of both sexes were represented equally but juveniles were rarely detected. High rates of translocation of adult toads of both sexes suggest that the eventual distribution of cane toads in Australia may be limited by the animals' bioclimatic tolerances rather than by an inability to reach suitable habitats, even in areas far distant from the toads' current range.     

Radial astrocytes and ependymocytes in the spinal cord of the adult toad (Bufo bufo L.)

Summary Immunohistochemical and ultrastructural techniques have been used to demonstrate glial fibrillary acidic protein (GFAP) immuno-positive cells in the adult toad spinal cord. Two types of GFAP-immunoreactive cells were observed: ependymocytes and radial astrocytes. GFAP-positive ependymocytes were scarce and contained the immunoreactive product in their processes. They showed intermediate filaments in the basal pole and in their processes when studied with the electron microscope. These immuno-positive ependymocytes represent the tanycytic form of ependymal cells because their processes ended at the subpial zone. The radial astrocytes showed a more intensive immunoreactive product in somata and processes when they were located far away from the ependymal layer. Cell bodies and processes were also associated with blood vessels, but most of the processes ended at the subpial zone forming a continuous subpial glia limitans. The GFAP-positive processes, which form this subpial glia limitans in the toad spinal cord, belong to both tanycytic ependymocytes and radial astrocytes, whose somata are located in the grey matter. These findings lead us to suggest that both types of GFAP-immunopositive cells might be the functional equivalents of mammalian astrocytes.     

L'ultrastructure de l'adénohypophyse du crapaud Bufo bufo L.

Résumé Cinq types de cellules granulées se différencient au cours de la métamorphose dans la pars distalis de l'hypophyse du têtard de Crapaud.A la prémétamorphose apparaissent les deux types de cellules protidiques, les cellules glycoprotidiques de type II et des cellules glycoprotidiques d'un type intermédiaire. Cette dernière catégorie cellulaire disparaît au climax.A la prométamorphose se différencient les cellules glycoprotidiques de type IV.Au climax s'observent les cellules glycoprotidiques de type III.Les cellules glycoprotidiques de type II sont vraisemblablement responsables de la sécrétion de l'hormone thyréotrope (TSH). Il n'est pas encore possible de préciser la fonction des autres types cellulaires.Des fibres nerveuses ont pu être observées dans la pars distalis entre les cellules granulées.

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The ultrastructure of the adenohypophysis in the toad, Bufo bufo L.III. Cell differentiation in the pars distalis during larval growth

Summary Five granulated cell types can be distinguished in the Toad's pars distalis during larval growth.During premetamorphosis the two types of protidic cells appear, the glycoprotein containing cells of type II and an intermediary cell type which disappears during the climax.During prometamorphosis the glycoprotein cells of type IV are apparent.During the climax the glycoprotein cells of type III can be observed.The glycoprotein containing cells of type II probably produce the thyroid-stimulating-hormone (TSH). The function of the other cell types can not be specified for the moment.Nervous fibers have been observed in the pars distalis between granulated cells.

Tous nos remerciements vont à Mme Sidler-Ansermet photographe, Mlle Schorderet secrétaire, et Mlle Schutz technicienne, de l'aide qu'elles ont apportée à la réalisation de ce mémoire.