Cytochemistry and polarization microscopy of amphibian sperm cell nuclei presumed to differ in basic protein composition

The pattern of availability of free DNA phosphates, and the kind of DNA-protein complex arrangement, both induced by nuclear basic proteins, and the richness in arginine residues in these proteins were investigated cytochemically and cytophysically in spermatozoa of the South-American Hylidae species, Hyla fuscovaria and Hyla biobeba. The aim was to demonstrate differences at the level of sperm histones in two species of Hyla until recently considered to be congeneric. The results indicated differences in the spermatozoal nuclear basic proteins and DNA-protein complexes when the two species were compared. The spermatozoa of Hyla biobeba were assumed to be likely to contain a Bloch's ''type 3'' protein type (intermediate sperm basic protein), similarly to Hyla species of North and Central America. on the other hand, the data obtained for the spermatozoa of Hyla fuscovaria indicated that they contain a protamine or protamine-like protein, differing from Hyla biobeba and Hyla species of North and Central America. It is suggested that the differences reported here may be genus-specific, since Hyla fuscovaria has recently been reclassified as Scinax fuscovaria based on parameters other than sperm histone types. These findings are in agreement with the general view of a wide variability in sperm nuclear proteins in the Anura group.

A NEW SPECIES OF HYLODES FROM SOUTHEASTERN BRAZIL (AMPHIBIA, LEPTODACTYLIDAE)

A new species of leptodactylid frog is described from the Serra das Cabras in São Paulo, southeastern Brazil. The new species is a member of the Hylodes lateristrigatus group and is characterized by its small size and high number of notes per call. Descriptions of the tadpole and advertisement call and information on natural history are provided.

A new species of Physalaemus (Amphibia; Leptodactylidae) from the Atlantic forest in southeastern Brazil

A new species of leptodactylid frog, Physalaemus atlanticus, is described from Ubatuba, northern coast of São Paulo in southeastern Brazil. The new species belongs to the Physalaemus signifer species group and is characterized by the following set of characters: small size (males 20.1-22.1 mm SVL, females 21.0-23.9 mm SVL); canthus rostralis distinct; dorsal skin texture smooth to slightly rugose; belly orange in life; advertisement call with duration of 0.6-0.84 s and frequency between 0.9-1.8 kHz. Descriptions of the advertisement call and tadpole are provided, as well as information on natural history.

HYLA-LUCTUOSA, A NEW TREEFROG FROM SOUTHEASTERN BRAZIL (AMPHIBIA, HYLIDAE)

A new species of treefrog, Hyla luctuosa, is described from the Serra do Japi in southeastern Brazil. The new species is a member of the Hyla circumdata group characterized by large size, large tympanum, and rounded subarticular tubercles on the fingers. Descriptions of the tadpole and advertisement call and information on natural history are provided.

Knowledge gaps and bibliographical revision about descriptions of free-swimming anuran larvae from Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

An austral anuran assemblage in the Neotropics: seasonal occurrence correlated with photoperiod

The study describes habitat use and temporal occurrence of calling males for an austral anuran assemblage in the Neotropics, southern Brazil. Three study sites (S1 - a permanent river and a stream, S2 - a dam and backwater, S3 - two permanent ponds periodically connected) were sampled between August 2005 and July 2006. The site S3 presented the richest and most diverse assemblage and habitat use within studied sites, and was partitioned by species groups. Richness and abundance of calling males for overall assemblage were seasonal, concentrated in spring and summer and correlated with photoperiod. Temperature, rainfall and air humidity were not correlated with the richness and abundance of calling males. Photoperiod also explained the calling seasons when species were analyzed individually. Habitat use and temporal occurrence were complementary in the partitioning of breeding resources, explaining species coexistence.

Investigation of the cytocidal potential of Rhinella jimi skin methanol extracts

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

New species of Hylodes from the Atlantic forest of Brazil (Amphibia: Leptodactylidae)

A new species of leptodactylid frog is described from Eldorado, state of Sao Paulo, southeastern Brazil. The new species is a member of the Hylodes lateristrigatus group and is characterized by its large size, slightly rugose to rugose dorsal skin texture, robust body, and high number of notes per call. Descriptions of the advertisement call and information on natural history are provided.

The thrombocyte aggregation process in the turtle Phrynopys hilarii (Chelonia). An ultrastructural study

This study investigates the thrombocyte aggregation process in the South American fresh water turtle (Phrynopys hilarii) using electron microscopy. Blood was taken from surgically exposed lateral neck vessels often turtles Phrynopys hilarii during the spring and summer seasons, when the mean temperature is 37°C. Blood samples were fixed with Karnovsky solution for processing by transmission electron microscopy. The turtle thrombocytes were spindle-shaped with lobulated nuclei. Prominent vesicles and canaliculi were found throughout the cytoplasm. The cytoplasm organelles showed an agranular endoplasmatic reticulum, Golgi complex near the centrioles and scattered free ribosomes. These cells are similar to bird thrombocytes but distinct from fish and frog thrombocytes. Blood clotting time was 5 min ± 30 sec measured by the Lee and White method. Structural alterations resulting from the aggregation process occurred after activation. Thrombocytes developed numerous filopodial projections, an increased number of vacuoles and changed from spindle to spherical shape. P. hilarii thrombocytes have different morphologic characteristics compared to other non-mammalian vertebrate cells. These cells can participate in the aggregation process, as observed in birds.

Testes size in leptodactylid frogs and occurrence of multimale spawning in the genus Leptodactylus in Brazil

Although rare, some anurans show multimale spawning. Herein we present data on testes mass relative to body mass for 11 leptodactylids and report multimale spawning in Leptodactylus chaquensis and Leptodactylus podicipinus. Testes mass averaged 4.13% of body mass for L. chaquensis and 0.75% for L. podicipinus. Testes size of both leptodactylid species with multimale spawning show that they are large and close to values recorded for rhacophorid frogs with multimale breeding (0.71-7.79%). Large testes size in frogs with multimale spawning supports the sperm competition hypothesis.

Testicular morphological changes during the seasonal reproductive cycle in the bullfrog Rana catesbeiana

Spermatogenesis and steroidogenesis undergo seasonal variations during the reproductive cycle in amphibians. Testicular morphological and morphometric seasonal variations as well as interstitial lipidic inclusions and intralobular glycoconjugates were evaluated during seasonal cycle of Rana catesbeiana. Testes of frogs collected during the annual seasons were weighed for calculation of GSI (Gonadosomatic index). Seminiferous lobule diameters (DSL) and volume densities of seminiferous lobules (VvSL), excretory ducts (VvED), and interstitial tissue (VvIT) were analyzed. Semithin sections were submitted to Periodic Acid-Schiff (PAS) and Alcian Blue (AB) methods for detection of glycoconjugates, while lipidic inclusions were detected by Sudan Black B. GSI showed no significant variations during the year. Since VvED and VvIT increased significantly during summer and were inversely proportional to VvSL, a compensatory effect between the testicular compartments may be related to the maintenance of GSI. During autumn/winter, larger lobular diameters were observed in comparison to spring/summer when spermiogenesis and spermiation were commonly observed. The increased VvIT and the numerous lipidic inclusions in the interstitial cells during summer suggest a relationship between spermiogenesis and steroidogenesis. Besides the structural stability variations occurring in the IT and SL, a possible paracrine interaction between ED and IT should be also involved in the IT development during summer. The presence of PAS and AB-positive globular structures were observed in the seminiferous lobules and excretory ducts. These structures containing acid glycoconjugates appear to be Sertoli cell apical portions, which are accumulated in the lumen of the seminiferous lobules mainly during spermiation. © 2004 Wiley-Liss, Inc.

Apparent digestibility of different ingredients in diets for bullfrog Rana catesbeiana tadpoles

The objective of this study was to determine apparent digestibility of protein and energy of ingredients commonly present in rations fed to bullfrog tadpoles Rana catesbeiana such as corn, soybean meal, wheat meal, rice bran, poultry by-product meal, blood meal, fish meal, corn starch and soybean oil. A completely randomized design was used to test ten experimental diets with four replicates. About 30% of the reference diet was replaced by the test ingredients, except for soybean oil that was used at 10% ratio and 0.5% of chromium oxide as inert marker. The values of apparent digestibility coefficient of crude protein (ADCCP) and gross energy (ADCGE) of the protein ingredients were, respectively: fish meal, 81 and 95%; poultry byproduct meal, 82 and 87%; blood meal, 64 and 53%; and soybean meal, 93 and 90%. While for the energetic ingredients, ADCCP and ADCGE, were, respectively: wheat meal, 100 and 72%; rice bran, 57 and 24%; and corn, 84 and 92%. Soybean oil and cornstarch ADCGE values were 70 and 96%, respectively; cornstarch digestibility is highly dependent on feed processing technique. The results showed that the ing redients of plant origin, except for rice bran, had good digestibility, thus indicating that tadpoles can effectively use plant protein. In this trial, the least digestible ingredients were blood meal and rice bran, thus indicating that the use of such ingredients in bullfrog tadpole diets should be limited. © Copyright by the World Aquaculture Society 2005.

Reproductive modes in frogs and their unexpected diversity in the Atlantic forest of Brazil

Amphibians, and especially the anurans (frogs and toads), exhibit a greater diversity of reproductive modes than other tetrapod vertebrates. Twenty-nine reproductive modes have been recognized for the anurans; we propose 10 more, elevating by more than 34% the number of reproductive modes known for anurans worldwide. These newly recognized reproductive modes for the frogs elevate by almost 48% the number of anuran reproductive modes known for the Neotropics. The highly complex topography of the Atlantic forest, breaking up the biome into many small microhabitats, and the high humidity, which reduces desiccation risks, have enabled the evolution of reproductive specializations such as eggs or tadpoles that develop out of water. Nearly 90% of the Atlantic forest has been cleared, and because several anurans are endemic to this region or have specialized reproductive modes dependent on the forest, this partly explains the generalized population declines and large numbers of species that have disappeared in the last few decades. © 2005 American Institute of Biological Sciences.