Electron microscopy study of lizard spermiogenesis in Tropidurus torquatus (Lacertilia)

In the course of an investigation on lizard spermiogenesis, the authors have observed events connected with the differentiation of the acrosome and middle-piece of the spermatozoa. Some differences from other reptilians were found as well as: the presence of a subacrosomal space, the absence of the fibrous sheath in the middle-piece, and a very short neck in the mature spermatozoa.

Food and feeding habits of the neotropical river otter Lontra longicaudis (Carnivora, Mustelidae)

A study of the feeding habits of the neotropical river otter, Lontra longicaudis, based on focal analysis was carried out from July 1986 to July 1987. The study was conducted at a dam in the 'Duas Bocas' Biological Reserve near the town of Cariacica, Espirito Santo State, Brazil. This reserve spreads over an area of 2 910 ha, the surface being mainly covered by the Atlantic Rain Forest. In order to identify the fish and other animal remains found in otter fecal samples, these were compared to the homologous structures of identical species living in the dam. The results of 288 samples were expressed in numbers and frequency of occurrence. Fishes were the most important food item, being present in 281 samples (97.2 %). A species of the genus Geophagus was frequently found in spraints, eaten during all months studied and present in 88.9 % of the samples. Astyanax, Pimelodella, Hoplias, Leporinus, Rhamdia, Tilapia and two other unidentified genera were found less frequently. In decreasing order of occurrence, crustaceans, amphibia, mammals, insects and birds were also encountered. Seasonal variation was verified in the scats between the dry and rainy seasons, with a higher frequency of food items occurring in the latter. In agreement with other species of otters, fishes also are the most commonly ingested prey. Two characteristics of fishes, greater abundance and easier capture, make them the major prey of otters, implying that otters are opportunistic predators eating whatever is more available.

Allometry and morphological integration in the skull of Tupinambis merianae (Lacertilia: Teiidae)

The morphogenetic processes acting in the skull of the lizard Tupinambis merianae were investigated by geometric morphometric techniques. The observed ontogenetic shape change involved a widening of the anterior extremity, stretching and narrowing of the midface, narrowing of the braincase, orbital reduction and elongation of the temporal region (origin of jaw adductor muscles). This change occurred mostly in a localized way in certain cranial regions. The major components identified were: rostrum, midface, dermal elements of braincase (functionally influenced) and endochondral elements of braincase (embryologically influenced). The growth patterns lead to an increased robustness of the skull (particularly the anterior extremity) and a reduction of cranial kinesis. These changes, together with the ontogenetic variation in dentition aid in the ontogenetic variation observed in the diet of these animals, which shift from carnivory to omnivory.

Escalated aggressive behaviour and facultative parental care in the nest building gladiator frog, Hyla faber

Agonistic encounters and facultative parental care in Hyla faber were observed in two localities in southeastern Brazil. Maximum male density was 0.9 and 3.3 males/m2 in Campinas and Ribeirão Branco, respectively. Aggression was escalated and the highly variable aggressive calls were specific to each phase of the encounter. The last, more aggressive phases rarely occurred in Campinas; in Ribeirão Branco they occurred frequently. Male parental care (egg attendance) was common in Ribeirão Branco while it was never observed in Campinas. Egg attendance lasted one to two nights and was observed only during high male density. The main benefit of egg attendance seemed to be avoiding nest intrusion by other males (sunken eggs and/or embryos invariably die). Males may build additional nests during egg attendance, but attending males did not attract females (they did not call).

Cytotaxonomic considerations with the description of two new NOR locations for South American toads, genus Bufo (Anura: Bufonidae)

Cytogenetic studies were made on Brazilian Bufo: B. marinus, B. paracnemis, B. ictericus, B. rufus, B. arenarum, B. crucifer, Bufo granulosus, B. pygmaeus and B. margaritifer (= B. typhonius). All these species had a typical karyotype of 2n = 22. Species from the marinus and crucifer groups had NORs on Chromosome 7, species from the granulosus group had NORs on Chromosome 5, and B. margaritifer had NORs on Chromosome 10. The last two locations of NORs are described for the first time for the genus Bufo in South America.

Cytogenetics of two species of Paratelmatobius (Anura: Leptodactylidae), with phylogenetic comments

In this paper we provide a cytogenetic analysis of Paratelmatobius cardosoi and Paratelmatobius poecilogaster. The karyotypes of both species showed a diploid number of 24 chromosomes and shared some similarity in the morphology of some pairs. On the other hand, pairs 4 and 6 widely differed between these complements. These karyotypes also differed in their NOR number and location. Size heteromorphism was seen in all NOR-bearing chromosomes of the two karyotypes. In addition, both karyotypes showed small centromeric C-bands and a conspicuous heterochromatic band in the short arm of chromosome 1, although with a different size in each species. The P. cardosoi complement also showed other strongly stained non-centromeric C-bands, with no counterparts in the P. cardosoi karyotype. Chromosome staining with fluorochromes revealed heterogeneity in the base composition of two of the non-centromeric C-bands of P. cardosoi. Comparison of the chromosomal morphology of these Paratelmatobius karyotypes with that of P. lutzii showed that the P. poecilogaster karyotype is more similar to that of P. lutzii than P. cardosoi. These cytogenetic results agree with the proposed species arrangements in the P. cardosoi and P. lutzii groups based on morphological and ecological data.

Histoenzymological and ultrastructural changes in lateral muscle fibers of Oreochromis niloticus (Teleostei: Cichlidae) after local injection of veratrine

The effects of veratrine have been investigated in mammalian, amphibian, and crustacean muscle, but not in fish. In this work, the action of veratrine was studied in the lateral muscle of the freshwater teleost Oreochromis niloticus after intramuscular injection. Histoenzymological typing and electron microscopy of muscle fibers before and 15, 30, and 60 min after veratrine injection (10 ng/kg fish) were used to indirectly assess the morphological changes and the oxidative and m-ATPase activities. In some cases, muscles were pretreated with tetrodotoxin to determine whether the ultrastructural changes were the result of Na+ channel activation by veratrine. Veratrine altered the metabolism of fibers mainly after 30 min. Oxidative fibers showed decreased NADH-TR activity, whereas that of glycolytic and oxidative-glycolytic type fibers increased. There was no change in the m-ATPase activity of the three fiber types, except at 60 min postveratrine, when a novel fiber type, which showed no reversal after acidic and alkaline preincubations, appeared. Ultrastructural damage involved sarcomeres, myofibrils, and mitochondria, but the T-tubules remained intact. Pretreatment with tetrodotoxin (1 ng/ml) prevented the ultrastructural changes caused by veratrine. These results show that in fish skeletal muscle veratrine produces some effects that are not seen in mammalian muscle.

Structural characterization of nuclear phenotypes during Scinax fuscovarius spermatogenesis (Anura, Hylidae)

In anuran amphibian Scinax fuscovarius, the spermatogenesis occurs in structures called seminiferous loculi, in which germ epithelium is organized in spermatocysts. Each cyst contains cells in the same stage of cytodifferentiation. Characteristics of each cellular type and their groups made the identification and differentiation of the germ lineage cells possible. In the basis of the epithelium there are the spermatogonia I, the biggest cells and always associated with the Sertoli cell. After the phase of mitotic proliferation, the cysts containing variable number of spermatogonia II are originated, quite smaller and with cellular boundaries a little distinct. After differentiation and growth in volume, the spermatocytes I appear, the nuclei of which are spherical and with different degrees of compaction of the nuclear material. Starting the meiotic process, the spermatocytes II are originated, which by means of the second meiotic division become haploid cells, the spermatids I. These two last spermatocysts are very similar. In this phase, the cells will go through a prominent process of differentiation until they form the spermatids II, which are elongated and begin to be organized in bundles supported by prominent Sertoli cells. With the process of spermiogenesis, spermatozoa appear, usually observed in compact bundles with tails turned to the lumen and their heads fitted in their support cells. In more advanced stages, the spermatozoa can be observed free in the locular lumen, ready to follow the spermatic path.

New species of Phrynohyas from Atlantic rain forest of southeastern Brazil (Anura, Hylidae)

A new species of hylid frog is described from south of the State of São Paulo, southeastern Brazil in Atlantic rain forest. The new species is a member of the genus Phrynohyas and is characterized by snout rounded in dorsal view and nearly truncate in lateral view, small adhesive disks, fingers slightly webbed, nasal bones widely separated, and frontoparietals without medial articulation.

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.

Visual signaling and reproductive biology in a nocturnal treefrog, genus Hyla (Anura: Hylidae)

We investigated reproductive activity, courtship behavior, call structure, body size, clutch size, oviposition site, and reproductive mode of Hyla sp. (aff. ehrhardti). Males called in all months, but showed a peak of activity during the rainy season. Three pair formations were observed and courtship involved stereotyped behavioral sequences, including visual signaling. Males emitted three different vocalization types: advertisement calls, courtship calls, and a vocalization of unknown function. Females attained larger body sizes than males and deposited an average of 74.5 unpigmented eggs per clutch. Early larval stages are aquatic but restricted to water in constructed subterranean nests; subsequent to flooding of nests, exotrophic tadpoles live in ponds or streams. Courtship behavior in Hyla sp. (aff. ehrhardti) is complex and the diverse repertoire of limb movements, exhibited by males, probably provide visual stimuli to females in this nocturnal treefrog. Hyla sp. (aff. ehrhardti) belongs to the H. albomarginata group. Considering the reproductive modes in this group, the complexes of H. albosignata and of H. albofrenata can be considered more close related to each other than to the H. albomarginata complex. © Koninklijke Brill NV, Leiden, 2004.

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.

Strong selection against hybrids at a hybrid zone in the Ensatina ring species complex and its evolutionary implications

The analysis of interactions between lineages at varying levels of genetic divergence can provide insights into the process of speciation through the accumulation of incompatible mutations. Ring species, and especially the Ensatina eschscholtzii system exemplify this approach. The plethodontid salamanders E. eschscholtzii xanthoptica and E. eschscholtzii platensis hybridize in the central Sierran foothills of California. We compared the genetic structure across two transects (southern and northern Calaveras Co.), one of which was resampled over 20 years, and examined diagnostic molecular markers (eight allozyme loci and mitochondrial DNA) and a diagnostic quantitative trait (color pattern). Key results across all studies were: (1) cline centers for all markers were coincident and the zones were narrow, with width estimates of 730 m to 2000 m; (2) cline centers at the northern Calaveras transect were coincident between 1981 and 2001, demonstrating repeatability over five generations; (3) there were very few if any putative F1s, but a relatively high number of backcrossed individuals in the central portion of transects: and (4) we found substantial linkage disequilibrium in all three studies and strong heterozygote deficit both in northern Calaveras, in 2001, and southern Calaveras. Both linkage disequilibrium and heterozygote deficit showed maximum values near the center of the zones. Using estimates of cline width and dispersal, we infer strong selection against hybrids. This is sufficient to promote accumulation of differences at loci that are neutral or under divergent selection, but would still allow for introgression of adaptive alleles. The evidence for strong but incomplete isolation across this centrally located contact is consistent with theory suggesting a gradual increase in postzygotic incompatibility between allopatric populations subject to divergent selection and reinforces the value of Ensatina as a system for the study of divergence and speciation at multiple stages. © 2005 The Society for the Study of Evolution. All rights reserved.

Evaluating the karyotypic diversity in species of Hyla (Anura, Hylidae) with 2n = 30 chromosomes based on the analysis of ten species

Ten species of Hyla with 2n = 30 from Brazilian fauna were analysed cytogenetically. Hyla minuta is the unique presenting all bi-armed metacentric or submetacentric chromosomes in the karyotype, therefore, with the highest FN = 60. The remaining species have a variable number of uni-armed telocentric or subtelocentric chromosomes: H. cruzi, H. elianeae, and H. rubicundula with three pairs (FN = 54), H. berthalutzae, H. elegans, H. microps, and H. nana with four pairs (FN = 52), and H. nahdereri and H. sanborni with five pairs (FN = 50). The uni-armed elements are among pairs 5, 6, 7, 11, 14, and 15, which also appeared with metacentric or submetacentric morphology. The remaining chromosome pairs 1, 2, 3, 4, 8, 9, 10, 12, and 13 were never found to be telocentric or subtelocentric. AgNOR patterns are species-specific, the majority of the species exhibiting a single pair with AgNORs, with the exception of H. elegans and H. nana with more than one chromosome pair bearing this cytological marker. C banding was obtained in H. berthalutzae, H. cruzi, H. elegans, H. elianeae, H. microps, H. minuta, H. nahdereri, and H. nana, which showed positively stained centromeric heterochromatin. Our analysis confirms the great karyotypic diversity in the species of Hyla with 2n = 30, with no species sharing identical karyotypes.