Brazilian Journal of Biology

Treatment of human breast epithelial cells MCF-10F with 17-β-estradiol has been reported to result in E2-transformed cells which have given rise to highly invasive C5 cells that in turn generate tumors in SCID mice. From these tumors, various cell lines, among which C5-A6-T6 and C5-A8-T8, were obtained. Although different phases of the tumorigenesis process in this model have been studied in molecular biology and image analysis assays, no cytological data on apoptotic ratios and mitotic abnormalities have been established to accompany the various steps leading to 17-β-estradiol-treated MCF-10F cells to tumorigenesis. Here we detected that the apoptotic ratio decreases with the transformation and tumorigenesis progress, except for the tumor cell line C5-A8-T8, probably on account of its more intense proliferation rate and a more rapid culture medium consumption. Increased frequency of mitotic abnormalities contributed by triple- and tetrapolar metaphases, and by lagging chromosomes and chromosome bridges observed at the anaphase found by transformation and tumorigenesis progress. However, no difference was found under these terms when the C5-A6-T6 and C5-A8-T8 tumor cell lines were compared to each other. Present findings are in agreement with the nuclear instability and enrichment of dysregulated genes in the apoptotic process promoted by transformation and tumorigenesis in 17-β-estradiol-treated MCF-10F cells.

In the lakes of the Middle Rio Doce, Minas Gerais (MG), two groups of larval Libellulidae are distinguished by preferences of habitat use: one uses mainly aquatic macrophytes and the other uses the bottom substrate. The goal of this work was to verify if there is a morphological distinction between the two groups of species. Thirteen body measures were taken from the larvae and analyzed. No difference was found between the two groups of species regarding the body size, but shape differences were observed for two morphological variables. The species that use mainly macrophytes tend to have larger relative measures of the labium and smaller measures of the abdomen width. Advantages in resource obtainment and in vulnerability to predation are probably the explanations for the morphological divergence among these larval groups.

The red hybrid tilapia, Oreochromis niloticus (Linnaeus, 1758) x O. mossambicus (Peters, 1852) is a fertile hybrid used in the semi-intensive level of fish culture in the Northeast of Brazil. It is a territorial cichlid and is highly aggressive towards conspecifics during the breeding season. The purpose of this study was to investigate and describe the aggressive behaviour displayed by the males of this hybrid in non-reproductive and reproductive contexts. Behavioural observations revealed that aggression displayed by the reproductive males of red hybrid tilapia included threatening, undulation, parallel, lateral and frontal attacks, chasing, escape and submission. Possession of a territory influenced male aggressiveness, which was more intense in their own territory than that observed in a neutral situation. The males built nests, irrespective of female presence. All the behavioural patterns were in accordance with those previously described for one parental species, the Nile tilapia, O. niloticus.

Causes and implications of sexual dimorphism have been studied in several different primates using a variety of morphological characters such as body weight, canine length, coat color and ornamentation. Here we describe a peculiar coat color characteristic in the squirrel monkey that is present only in adult females over five years old and which increases with age. Neither males nor young animals manifest this phenomenon, which is characterized by a spot of black hairs located anteriorly to the external ear (pinna). This characteristic could be used to discriminate adult females of Saimiri sciureus in the wild without the need of capture techniques.

We studied the role of white-lipped and collared peccaries (Tayassu pecari and Pecari tajacu) as seed predators and dispersers in the Atlantic rainforest of Brazil. The Atlantic rainforest ecosystem is highly threatened and has experienced dramatic declines in its populations of large mammals. Local extinctions can disrupt essential plant-animal interactions such as seed dispersion and seed predation. We tracked seeds from time of consumption to germination to assess the direct impact peccaries have on seed survival. We offered fruits of 20 species found in the Atlantic rainforest to the peccaries. Seeds were categorised as intact, scarified, ingested or defecated, and germination tests were performed. The overall impact by both peccary species was similar. Seeds were sometime scarified by mastication, always with fatal consequences. Most seeds that were consumed were destroyed during ingestion and digestion. Only small seeds (<10 mm) were found in the feces and germination tests suggest a positive effect from the passage through the guts. Peccaries clearly have a double role as both seed predators and as small seeds dispersers, which is a specialised role within the granivore/frugivore community of the Atlantic rainforest.

Teleostean fins when partially amputated suffer a regenerative process called epimorphic regeneration, characterized by the following stages: healing, based on the formation of a multistratified epidermal layer, the formation of a mass of pluripotent cells known as blastema, the differentiation of these cells, the synthesis and disposition of the extracellular matrix, morphological growth and restoration. The epidermis has a fundamental role in the regenerative process of fish fins, as the healing time of this structure leads it to a faster regenerative process and it also works as a defense against the external environment. In this sense, due to the fast regeneration shown by the epidermis, the aim of this paper is to study the histology of the regenerative dynamics of the carp fin tail (Cyprinus carpio), under the light and transmission electron microscope. Epidermic regeneration begins right in the first hours after the fin amputation and it continues throughout the regenerative process. After 24 hours, an apical epidermal cap is established. Cytoplasmatic prolongations and intercellular junctions are observed and the cells of the basal layer of the epidermis change from the cubic form to the cylindrical, due to the development of the cytoplasmatic organelles responsible for the synthesis of the basal membrane, lost after amputation. These results show the importance of histological studies in regenerative processes. We believe that the association of molecular biology with histological studies can throw further light onto these regenerative dynamics.

Physiological studies of the mussel Perna perna in Brazil are almost 30 years behind those of other, more exhaustively investigated species, such as Mytilus edulis. Little is known about the variations in physiological rates due to size and the consequences of maintaining P. perna in laboratory conditions. This work investigated the variations in respiration, clearance, excretion and absorption efficiency rates of P. perna, classified by size and acclimatized in a laboratory, monitoring the mussels respiration rates and biometry over a period of 30 days, in laboratory conditions. The respiration, clearance and excretion rates presented an allometric relation with the dry weight of the organisms, with b values of 0.66, 0.48 and 0.91 respectively. On the other hand, these same rates, when considered by weight (specific rates) showed a relationship that was inverse to the size of the organisms. Only the absorption efficiency was independent of the weight of the mussel. In terms of acclimatization, it was observed that it takes 10 days for the respiration rate of the mussel P. perna to stabilize in laboratory conditions, after which it follows a routine metabolism.

Chelonia mydas is a sea turtle that feeds and nests on the Brazilian coast and a disease called fibropapillomatosis is a threat to this species. Because of this, it is extremely necessary to determine a methodology that would enable the analysis of blood leukocyte function in these sea turtles. In order to achieve this aim, blood samples were collected from C. mydas with or without fibropapillomas captured on the São Paulo north coast. Blood samples were placed in tubes containing sodium heparin and were transported under refrigeration to the laboratory in sterile RPMI 1640 cell culture medium. Leukocytes were separated by density gradient using Ficoll-PaqueTM Plus, Amershan Biociences. The following stimuli were applied in the assessment of leukocyte function: Phorbol Miristate-Acetate (PMA) for oxidative burst activity evaluation and Zymosan A (Saccharomyces cerevisiae) Bio Particles, Alexa Fluor 594 conjugate for phagocytosis evaluation. Three cell populations were identified: heterophils, monocytes and lymphocytes. Monocytes were the cells responsible for phagocytosis and oxidative burst.

The presence of pollutants in the ocean may affect different physiological parameters of animals. Oxygen consumption and ammonia excretion were evaluated in D-shaped larvae of mussels (Perna perna) exposed to zinc sulphate (ZnSO(4)) and benzene (C(6)H(6)). When compared to the control group, both pollutants presented a significant reduction in oxygen consumption. A reduction in the ammonia excretion was also observed, both for ZnSO(4) and C(6)H(6) and also in the oxygen consumption. The results indicate that anaerobic metabolism may occur at the beginning of P. perna mussels development, as observed in veliger larvae. The O:N ratio under experimental conditions showed low values indicating that catabolism in veliger larvae was predominantly proteic.

The Pampas deer (Ozotoceros bezoarticus) is one of the most endangered Neotropical cervid with populations that have been drastically reduced to small and isolated ones, mainly because of its habitat destruction. Random amplified polymorphic DNA (RAPD) markers were used to analyze population divergence and genetic variation within and between two populations corresponding to distinct subspecies. The RAPD markers displayed substantial genetic variation with all animals possessing unique RAPD phenotypes over 105 polymorphic bands produced by 15 primers. An analysis of molecular variance (AMOVA) and a neighbor-joining cluster analysis were performed to assess levels of differentiation between populations. No differentiation was recorded and about 96.0% (P<0.00001) of the total variance was attributable to variation within populations. This result is quite distinct from data obtained by the analysis of the mtDNA control region, and is discussed on the basis of genetic differences between the different markers and the male-biased dispersal patterns generally observed in the mammal species. The data presented herein are potentially useful for future taxonomic and genetic studies in this species, for the monitoring of the genetic variation observed within these populations, and for the development of management guidelines for its conservation.

Oxygen of tropical freshwater environments fluctuates drastically. Eutrophic lakes and ponds of warm waters frequently reach very low oxygen concentrations. This is the most common habitat of the banded knife fish "tuvira" Gymnotus carapo. This electric fish is reported to present bimodal breathing to cope with low environmental oxygen. Biochemical responses can be also observed in fishes facing hypoxia but none were studied in tuvira. In the present study, haematological and metabolic changes were investigated in two groups of fish exposed to hypoxia for 1 and 3 hours. Haematocrit, red blood cells and haemoglobin concentration indicated erythrocyte release from hematopoietic organs and swelling of red blood cells. Glycogen, glucose, lactate, pyruvate, and amino acids were quantified in liver, kidney and white muscle. The metabolic profile of G. carapo to cope with hypoxia suggested liver gluconeogenesis probably supported by proteolysis. The kidney and liver presented the same biochemical trend suggesting similar metabolic role for both organs. Glucogenolysis followed by glucose fermentation and protein mobilisation was observed in the white muscle. The air breathing behaviour of tuvira works in parallel with metabolism to prevent damages from hypoxia. Metabolic adjustments are observed when the air taking is avoided.

Armadillos of the species Dasypus novemcinctus have been used as an experimental model of leprosy. Besides non-human primates, they are the only species naturally infected with Mycobacterium leprae and when experimentally inoculated, reproduce the lepromatous form of the disease producing large quantities of bacilli. This species has been maintained in captivity by numerous researchers and specific housing and feeding requirements have been developed to guarantee their survival during long experimental periods. In the 'Lauro de Souza Lima' Institute, armadillos receive dog food, ground beef, boiled eggs and vitamin C. However, despite the balanced diet, anemia has been observed in some captive animals, especially in armadillos inoculated with M. leprae in advanced stages of infection. Thus, the objective of the present study was to evaluate the effect of iron sulfate supplementation in the feed provided for armadillos, both inoculated and non-inoculated with M. leprae, by means of the evaluation of their hematological profile. Fourteen armadillos received 10 mg/animal of iron sulfate (Hematofer(R)) diluted in sterile water mixed with their daily feed for 50 days. Hemograms and serum iron dosages for each armadillo were performed before and after supplementation. The hematocrit values increased significantly after iron supplementation, both in armadillos inoculated and non-inoculated with M. leprae. It is possible that the amount of iron in the feed is insufficient for the formation of hemoglobin, leading to microcytic anemia. Dietary supplementation with iron sulfate reversed this state, showing the importance of understanding the metabolism of exotic species for their maintenance in captivity, and thus ensuring their well-being.

In marine bivalve mollusks, unsaturated molecules called carotenoids are present in the natural diet and play an important role in different biological process, especially in reproduction. In order to gain more insights into these compounds in Nodipecten nodosus it was necessary to develop a suitable protocol for extraction of carotenoids from the gonads. Female gonads of cultured scallops (75 mm length) were lyophilized and macerated in liquid N2. To verify the effect of composition in organosolvents on the extracting solutions, two organic solvents were tested: acetone and hexane (Ac = O:Hex) at four ratios, 1:1, 1:3, 1:5, and 2:3, in four static extraction times: 0, 5, 10, and 15 minutes. Total carotenoids and astaxanthin contents were determined in the crude extracts by UV-visible spectrophotometry and high performance liquid chromatography (HPLC), respectively. Triplicate aliquots of 50 mg were used for each treatment. The results indicated that the best single extraction (0.312 +/- 0.016 microg carotenoids/mg) was attained with Ac = O: Hex 1:3, for 15 minutes. Through exhaustive extraction methodology (10x), a superior yield (0.41 +/- 0.001 microg carotenoids/mg) was obtained from a gonad sample in comparison to the highest value found for a single extraction. Astaxanthin content was reduced by 8.6% in carotenoid extract preservation assay, i.e., -18 degrees C, 26 days incubation, under N2 atmosphere.

Previous studies have shown the superior competitive ability of honeybees compared with native bees in the exploitation of floral resources and nesting sites besides their low efficiency in pollinating native plant species. However, there is little evidence of the effect of this invading species on autochthonous plant populations in natural environments. Thus experiments were performed to test the pollination efficiency of honeybees in two species of Jatropha (Euphorbiaceae), J. mollissima (Pohl) Baill. and J. mutabilis (Pohl) Baill., after a single flower visitation. Samplings were carried out between March and April 2006 in a hyperxerophilous shrub-arboreal Caatinga at Estação Biológica de Canudos, Bahia (9º 56´ 34" S, 38º 59´ 17" W), the property of Fundação Biodiversitas. Apis mellifera was efficient at pollinating J. mollissima (100%) and J. mutabilis (85%). This high efficiency may be explained by 1) the simple floral characteristics of both plant species, which facilitate access to the sexual organs of the plant; and 2) the body size of A. mellifera that fits the flower's dimensions.

A conglomerate of small, rigid, fusiform spicules known as actinotrichia sustains the edge of tail fins of teleost. After amputation, these structures show an extremely fast regenerative capacity. In this study we observed the effect of a nonsteroidal anti-inflammatory drug, naproxen, used in the treatment of degenerative articular diseases, during the process of actinotrichia regeneration. For this purpose, regenerating tissue from animals in contact with the drug was submitted to histochemical and ultrastructural analysis in comparison to tissue from animals under normal conditions, i.e., not in contact with the drug in question. Actinotrichia regeneration was similar in both animals, indicating that naproxen, at the dose used in the present study, did not interfere with actinotrichia synthesis during the regenerative process of the tail fin. This could be because naproxen did not influence the expression of the genes required for the regeneration process, such as the Sonic hedgehog (Shh) gene, which is involved in actinotrichia formation.

To investigate whether source proximity or bioavailability is the major factor controlling both Hg concentration and Hg speciation in marine fishes, total- and organic-Hg content in muscle and liver tissues from different populations of Cephalopholis fulva (piraúna) from inshore and offshore waters of the Brazilian northeastern coast were analyzed. Average total-Hg in muscle (104 ng x g(-1) w.w.) and liver (2,865 ng x g(-1) w.w.) tissues, as well as organic-Hg concentrations in muscle (169 ng x g(-1) w.w.) and liver (1,038 ng x g(-1) w.w.), were much higher in the offshore population of C. fulva than in the inshore ones. In the inshore population total-Hg and organic-Hg average concentrations in muscle tissue were similar and reached only 49 ng x g(-1) w.w., while in liver they reached 412 ng x g(-1) w.w. for total-Hg and 180 ng x g(-1) w.w., for organic-Hg. Concentrations of both Hg species in the two populations were higher in liver than in muscle. The average percentage contribution of organic-Hg to the total Hg content was higher in muscle (98-100%) than in liver (42-53%), but similar between the two populations. Total-Hg and organic-Hg concentrations in muscle and liver from the offshore population showed significant (P < 0.05) positive correlation with fish length. However, in the inshore population only the total-Hg and organic-Hg in muscle tissues correlate significantly with fish size. Although the coastal environments are enriched in total Hg relative to open waters, the significant higher Hg concentrations in the offshore population of C. fulva and the significant correlation found between organic-Hg in liver with fish size suggest higher bioavailability of Hg in offshore waters relative to inshore ones.

This work describes the gametogenic cycle of the scallop Nodipecten nodosus kept in a culture system. To this end, during one year, samples were taken from the broodstocks every 30 days to be submitted to macroscopic and microscopic analyses and to measure the amount of astaxanthin. To perform the microscopic evaluation, 5 micro slices from the median portion of the female part of the gonad were submitted to the pattern methodology for histological analyses with paraffin and HE coloration. The remaining portion of the female gonad was lyophilised to extract and quantify the levels of astaxanthin using HPLC. The microscopic analyses revealed four well defined stages for the reproductive cycle. Analyses of data taken throughout the year indicated preferential spawning periods from December to January and from July to September. The astaxanthin analyses showed higher amounts of this carotenoid during the advanced pre-spawning and the initial spawning periods than during gametogenesis, initial pre-spawning, advanced spawning, and the spent stages. According to these results, it was possible to establish a descriptive table of the sexual stages of the female portion of the gonad and the amount of astaxanthin in the sexual stage of the scallop Nodipecten nodosus.

Raillietiella gigliolii is a Neotropical pentastomid parasite found in Amphisbaena alba. Collections were made in northeastern Brazil in a remnant area of Atlantic Forest (07 masculine 10' S and 35 masculine 05' W) in the municipality of Cruz do Espírito Santo, Paraíba State, and in a Humid Forest area (07 masculine 16' S and 39 masculine 26' W) on the slopes of the Chapada do Araripe Mountains, municipality of Crato, Ceará state, Brazil. Nine specimens of A. alba and 12 of A. vermicularis were collected to gather basic ecological data (prevalence and mean intensity of infection) concerning these parasites. Raillietiella gigliolii was found infecting the lungs of both species. The prevalence for A. alba was 55.5% (5/9), with a mean intensity of infection of 5.0 +/- 2.53 and amplitudes of 1-13. A. vermicularis demonstrated prevalence of 50%, with a mean intensity of infection of 5.3 +/- 2.1 and amplitudes of 1-14. This represents the first record of R. gigliolii as a parasite of A. vermicularis. Our results suggest that R. gigliolii is a generalist parasite species and that an overlapping diet is the determinant factor in the sharing of its final hosts.

A general analysis of the behaviour of "Cebus" shows that when this primate moves position to feed or perform another activity, it presents different ways of locomotion. This information shows that the brachial biceps muscle of this animal is frequently used in their locomotion activities, but it should also be remembered that this muscle is also used for other development activities like hiding, searching for objects, searching out in the woods, and digging in the soil. Considering the above, it was decided to research the histoenzimologic characteristics of the brachial biceps muscle to observe whether it is better adpted to postural or phasic function. To that end, samples were taken from the superficial and deep regions, the inserts proximal (medial and lateral) and distal brachial biceps six capuchin monkeys male and adult, which were subjected to the reactions of m-ATPase, NADH-Tr. Based on the results of these reactions fibres were classified as in Fast Twitch Glycolitic (FG), Fast Twitch Oxidative Glycolitic (FOG) and Slow Twitc (SO). In general, the results, considering the muscle as a whole, show a trend of frequency FOG> FG> SO. The data on the frequency were studied on three superficial regions FOG=FG>SO; the deep regions of the inserts proximal FOG=FG=SO and inserting the distal FOG>FG=SO. In conclusion, the biceps brachii of the capuchin monkey is well adapted for both postural and phasic activities.