Ultramorphological changes in gill rakers of Astyanax altiparanae (Characidae) kept in contaminated environments.

Most water bodies in Brazil, and in the world, are contaminated by some types of pollutants, ranging from sewage to metal/chemicals, carcinogenic products, and biodegradable detergents. Despite the extensive knowledge on their effects on fish biology and especially on gill morphology, research that concerns their impacts on gill rakers and implications in parameters such as food consumption cannot be found in the literature. Gill rakers are vital because, together with gills, they are responsible for the defense and protection of the organism and for selecting appropriate food for survival. When detergents, which can act as toxic chemical agents, get in contact with the body of the fish, they can cause severe effects that must be understood. Therefore, our study investigated ultramorphological changes in gill rakers of Astyanax altiparanae (Lambeth) caused by the exposure to biodegradable detergents. Fish were exposed to a 1 mg/L dilution of a mixture of detergents and pure water from an artesian well for 5 months. Results revealed that the first month of exposure to detergent caused dilation of chemical receptors in taste buds and the rise of a large number of orifices for mucus release among pavement cells in gill rakers, although only a small amount of mucus was found in fish exposed both to pure water and the detergent dilution. After 5 months, there was an increase in the dilation of these chemoreceptors, excess mucus on gill rakers of detergent groups, and the emergence of microbridges between microridges in pavement cells.

The scale epithelium as a novel, non-invasive tool for environmental assessment in fish: Testing exposure to linear alkylbenzene sulfonate.

Increasing pollution levels have turned our attention to assessing lethal and sublethal effects of toxic agents using the most informative techniques possible. We must seek non-invasive or non-lethal sampling methods that represent an attractive alternative to traditional techniques of environmental assessment in fish. Detergents are amongst the most common contaminants of water bodies, and LAS (Linear Alkylbenzene Sulfonate) is one of the most used anionic surfactant on the market. Our study analyzed morphological alterations (histological and histochemical) of the scale epithelium of Prochilodus lineatus under exposure to two concentrations of LAS, 3.6mg/L and 0.36mg/L, for a period of 30 days and evaluated at 14, 21 and 30 days. In order to establish morphological analysis of the scale epithelium as a new non-lethal environmental assessment tool that is reliable and comparable to classic methods, the relative sensibility of this technique was compared to a commonly used method of environmental assessment in fish, the estimation of the effects of pollutants upon branchial morphology. Two experiments were carried out, testing animals in tanks, and in individual aquariums. Results of analyses on gill tissue show that exposure to 3.6mg/L of surfactant caused severe damage, including hyperplasia, hypertrophy and fusion at 14 days, with aneurisms at 21 and 30 days; while exposure to 0.36mg/L had lighter effects on the organ, mainly lower incidence of fusion and hyperplasia. Aditionally, scale morphology was altered severely in response to 3.6mg/L of LAS, consistently showing increased mucous and club cell production. Epithelial thickness was the most variable parameter measured. Scale epithelium sensibility has the potential to be a reliable environmental marker for fish species since it has the advantage of being less invasive when compared to traditional methods. However, more studies are required to increase the robustness of the technique before it can be generally applied.

Scale morphology of Prochilodus lineatus with emphasis on the scale epithelium.

The fish body is entirely covered by a thin, smooth and glandular epidermis, closely attached to the scales inserted on the dermis. The descriptive work on this tissue dates to twenty or thirty years ago, bears very little photographic record and does not focus on the scale epithelium, despite the fact that it is in direct contact with the environment. Thereupon, the present study characterizes the scale epithelium of Prochilodus lineatus, a robust species of fish. The observations show that the scale is completely covered by epithelium thicker on the proximal end of the scale, multilayered on the dorsal surface and undifferentiated on the ventral surface, and covered by mucous producing cells, mostly acid mucous. The scale is formed by plywood-like collagen matrix of collagen type III and supported by a network of elastic fibers on the ventral face. Differentiated cellular types are present, such as club cells, considered to be responsible for the release of alarm substances, which suggests possible use in environmental assessment as a non-invasive technique.

Histochemical and ultrastructural analysis of hepatic glycogen and collagen fibers in alloxan-induced diabetic rats submitted to long-term physical training.

Alterations in liver functions are common among diabetic patients, and many symptoms in the liver have been reported, including changes in glycogen stores and in the amount of collagen fibers. The practice of physical training and its morphological effects in this organ, however, are scarcely studied. In order to observe the morphological effects of alloxan-induced diabetes and the alterations arising from the practice of long-term chronic physical training in the liver, samples were collected and processed, and then analyzed by means of the histochemical techniques Periodic Acid-Schiff and Picrosirius-Hematoxylin, and ultrastructural cytochemical test of Afzelius. Through evaluation of the tissue, it was observed a drastic reduction in hepatic glycogen stores of sedentary diabetics, recovered in trained diabetic rats. Furthermore, it was detected a decrease in the content of perisinusoidal collagen fibers in the diabetic liver, also recovered due to the development of a training protocol. On ultrastructural level, cytochemical analysis confirmed the loss of glycogen and the recovery obtained by training. In conclusion, the practice of a long-term chronic physical training protocol may be considered an important assistant in the treatment of diabetes, mitigating the occurrence of possible damages to liver tissue.

Collagen and reticular fibers in left ventricular muscle in diabetic rats: physical exercise prevents its changes?

Diabetic cardiomyopathy contributes to the high incidence of mortality in both types of diabetes. We aimed to investigate the histochemical aspects of collagen and reticular fibers in the cardiac muscle and evaluate the influence of physical exercise on these aspects. Wistar rats were divided in 4 groups: sedentary control (SC), trained control (TC), sedentary diabetic (SD) and trained diabetic (TD). Diabetes was induced with alloxan (35 mg/kg). Training program consisted of swimming 1h/day with a load of 4.8% of body weight for TD and 5.2% for TC, during 8 weeks. At the end of the experiment, samples of the left ventricle were collected, fixed in Bouin and processed on historesin. Sections were stained with periodic acid of Schiff, picrosirius-hematoxylin and ammoniacal silver. The PAS technique shows that individuals of group SD presented more intense reaction that the other groups. Picrosirius-hematoxylin technique showed a possible deposition of collagen fibers in SD. The TD group presented a reaction a lot similar to the controls' for both techniques', showing a possible prevention of this deposition. These results indicate that physical exercises might have an important role on the prevention of some negative alterations caused by experimental diabetes.

The midgut of Cephalotes ants (Formicidae: Myrmicinae): ultrastructure of the epithelium and symbiotic bacteria.

The ultrastructural analysis of the midgut of Cephalotes atratus, C. clypeatus, and C. pusillus reveled that the midgut epithelium lays on a basal lamina and is composed basically of three cell types: digestive cells, regenerative cells, and goblet cells. In these ants, the rough endoplasmic reticulum, in addition to producing digestive enzymes, is involved in the formation of concretions and ion storage in specialized vacuoles present in the midgut. These concretions are spherocrystals and may contribute to stabilize the pH and to maintain symbiotic bacteria found between microvilli. The ultrastructure analysis of these bacteria revealed the presence of a double envelope typical of gram-negative bacteria. For the three species examined, the ultrastructure similarities are conspicuous, suggesting that this may be the pattern for the genus Cephalotes. Details of the relationship between bacteria and microvilli were examined.

Molecular and ultrastructural profiles of the symbionts in Cephalotes ants.

The molecular and ultrastructural profiles of the symbionts found in the midgut and ileum of Cephalotes atratus, Cephalotes clypeatus, and Cephalotes pusillus were determined using the V3 region of the bacterial 16S rDNA gene and transmission electron microscopy (T.E.M.). Two samples of C. atratus, three of C. clypeatus, and six of C. pusillus were analyzed. The coefficients of similarity ranged from 80% to 94% for the samples of symbionts from C. clypeatus and C. atratus, despite being collected in geographically distant sites. The variability within symbionts found in the samples of C. pusillus varied from 29% to 55%, in samples geographically close as well as distant. PCR-DGGE was effective for the purpose of this study and can be considered a versatile tool to analyze gut microbiota. Details of the ultrastructural aspect of these bacteria are presented.

Intrinsic and extrinsic factors in the conservation of ants and their use as bioindicators / Fatores intrínsecos e extrínsecos na conservação de formigas e seu uso como bioindicadores

Ant communities are sensitive to fragmentation responding through: extinction of specialist species compared with large habitats, invasion of generalist species, changes in community composition and richness of species. Because of these characteristics, this review aims at examining the main risks that ants have been facing to maintain their communities, as well as to report current conservation practices in Brazil and in the world.

Comunidades de formigas são sensíveis à fragmentação e respondem por meio: da extinção de espécies especialistas comparadas com grandes habitats, invasão de espécies generalistas, mudanças na composição da comunidade e na riqueza de espécies. Por causa dessas características, esta revisão pretende descrever os principais riscos que as formigas têm enfrentado para manter suas comunidades, como também descrever práticas atuais no Brasil e no mundo.

Symbiotic bacteria and the structural specializations in the ileum of Cephalotes ants.

Light, scanning, and transmission electron microscopy were used to examine the ultramorphology and ultrastructure of the ileum of Cephalotes atratus, Cephalotes clypeatus, and Cephalotes pusillus. Sections along the ileum revealed differences among the three main regions: proximal (or pylorus), medial, and distal. The structural specializations present in the ileum of these three ants have implications especially to the symbiotic bacteria harbored in this region of the digestive tract. The structural similarities are conspicuous for the three species examined, suggesting that this is the pattern adopted by the genus Cephalotes.

Proventriculus of Cephalotes ants: a structural and comparative analysis.

The use of optic microscopy and scanning electron microscopy propitiated the comparative examination of the structure of the proventriculus bulb of Cephalotes atratus, Cephalotes clypeatus and Cephalotes pusillus. This portion of the digestive tract possesses highly sclerotized projections which act in the selection of victuals. This structure is of importance to phylogeny studies and thus was compared with others known species of ants in a dendrogram adapted from Eisner (1957). We did not detect differences among the three studied species in the nature of histochemistry and ultra morphology.

A new strategy of endosymbiont midgut bacteria in ant (Ponerinae).

The location and ultrastructure of bacteria associated with microvilli in the midgut of Odontomachus bauri were examined by transmission electron microscopy. These filamentous type bacteria are the second morphotype described in the midgut of this ant. They colonizes only the ectoperitrophic space, more specifically attaching along microvilli. A thick capsule attaches bacteria to microvilli and protect them from acidic pH and digestive enzymes. Details of the location and association with microvilli are discussed.

First report of endocytobionts in the digestive tract of ponerine ants.

We conducted ultrastructural studies to examine the presence of microorganisms in the midgut of the ant Odontomachus bauri (Ponerinae), as a contribution towards understanding the relationships between microorganisms and their hosts. The presence of microorganisms in this region, including inside the cells, suggests their participation in food digestion as symbiontic organisms and represent a new possibility to exploit food sources in the environment.

Contribution of the Malpighian tubules for the maintenance of symbiotic microorganisms in cephalotes ants.

Given the physiological importance of the Malpighian tubules to homeostasis in ants, this study aimed to characterize the enzymology, histology, histochemistry, and ultramorphology of the Malpighian tubules of Cephalotes atratus, C. clypeatus, and C. pusillus, as a contribution for the understanding of this organ, as well as to examine its role in the maintenance of symbiontic microorganisms in the ileum of these ants.

Ileum of the Cephalotes ants: a specialized structure to harbor symbionts microorganisms.

We conducted comparative morphological studies of the tract digestive of Cephalotes atratus, C. clypeatus and C. pusillus, aiming at describing the relationships among these three species, as well as the histochemical, enzymatic, and ultra-morphological differences of the wall and the contents of the digestive tract, therefore contributing for the understanding of the relationships between symbionts and their host.

Effects of swimming training on bone mass and the GH/IGF-1 axis in diabetic rats.

The aim of this study was to examine the influence of moderate swimming training on the GH/IGF-1 growth axis and tibial mass in diabetic rats. Male Wistar rats were allocated to one of four groups: sedentary control (SC), trained control (TC), sedentary diabetic (SD) and trained diabetic (TD). Diabetes was induced with alloxan (35 mg/kg b.w.). The training program consisted of a 1h swimming session/day with a load corresponding to 5% of the b.w., five days/week for six weeks. At the end of the training period, the rats were sacrificed and blood was collected for quantification of the serum glucose, insulin, GH, and IGF-1 concentrations. Samples of skeletal muscle were used to quantify the IGF-1 peptide content. The tibias were collected to determine their total area, length and bone mineral content. The results were analyzed by ANOVA with P<0.05 indicating significance. Diabetes decreased the serum levels of GH and IGF-1, as well as the tibial length, total area and bone mineral content in the SD group (P<0.05). Physical training increased the serum IGF-1 level in the TC and TD groups when compared to the sedentary groups (SC and SD), and the tibial length, total area and bone mineral content were higher in the TD group than in the SD group (P<0.05). Exercise did not alter the level of IGF-1 in gastrocnemius muscle in nondiabetic rats, but the muscle IGF-1 content was higher in the TD group than in the SD group. These results indicate that swimming training stimulates bone mass and the GH/IGF-1 axis in diabetic rats.

Histology and histochemistry of the ventriculus of Dolichoderus (=Monacis) bispinosus (OLIVIER, 1792) (Hymenoptera: Formicidae).

In this study we histologically and histochemically describe the ventriculus of Dolichoderus bispinosus. The epithelium consists of two basic cell types, highly basophilic generative cells, and digestive cells. The latter present several cytoplasmic vesicles, rich in acidic and neutral polysaccharides, and basic proteins. Also, these cells exhibit an apocrine secretion pattern. A mass of fibrous material is observed on the surface of the epithelium. Finally, we discuss the results obtained.

Ultramorphological analysis of the venom glands and their histochemical relationship with the convoluted glands in the primitive social paper wasp Polistes versicolor (Hymenoptera: vespiade)

The venom glands are part of the most important defense weapon in Aculeata: the venom apparatus. The arrangement of these glands can vary among species, but in general they are composed of long secretory tubules connected to a muscular sac-like reservoir. Although the occurrence of these variations has been documented, many studies neglected the existence of a well-developed secretory portion in the lumen of the reservoir named convoluted gland. This study is an ultramorphological analysis of the venom glands and their histochemical relationship with the convoluted glands in the primitive social wasp Polistes versicolor. In this wasp, the venom glands are constituted by two tubular portions tha penetrate individually in the venom reservoir, inside of which we can find the convoluted glands. Besides morphological differences in their cells, histochemical analysis of the venom and convoluted glands clearly show differences between them. While the venom glands indicate positive reaction only for proteins, the convoluted glands present positive reaction for proteins, neutral glycoconjugates, and lipids. The secretion of the convoluted gland cells may modify the compounds passing through the embedded tubular region

Ultramorphology and histochemistry of fat body cells from last instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae).

The fat body cells of Pachycondyla (=Neoponera) villosa are disposed in a single layer between the cuticle and the digestive tract, forming a group of cells enclosed by a thin membrane. Histological studies have revealed three different cellular types: trophocyte (more abundant), urate, (located among the trophocytes), and oenocyte (the scarcest), usually observed laying near the cuticle. Histochemically, the trophocytes showed a positive reaction for basic proteins in the nucleus and cytoplasm, as well as a strong positive reaction in the cytoplasmic granules. The test for carbohydrates showed a strong positive reaction throughout the cytoplasm, while the test for lipids was positive for the cytoplasmic vesicles. The urate cells showed a positive reaction for basic proteins in the nucleus and in the areas of the cytoplasm surrounding the vesicles. These cells did not react to the PAS test or to Sudan Black B. The oenocytes showed a weak positive reaction to PAS and a strong positive reaction to Sudan Black B and Mercuric-bromophenol Blue.

Ultramorphology and histochemistry of fat body cells from last Instar larval of the Pachycondyla (=Neoponera) villosa (Fabricius) (Formicidae: Ponerinae)

As células do corpo gorduroso de Pachycondyla (=Neoponera) villosa distribuem-se como uma única camada entre a cutícula e o trato digestivo, formando um conjunto de células agrupadas e recobertas por uma fina membrana. Não foram identificados tipos celulares distintos por meio da ultramorfologia, porém a histologia revelou três tipos celulares distintos: os trofócitos, mais abundantes, as células de urato, distribuídas por entre os trofócitos, e os enócitos, menos abundantes que os demais. Os enócitos são comumente observados próximos da cutícula. Histoquimicamente, os trofócitos apresentaram reação positiva para proteínas básicas no núcleo e no citoplasma e reação fortemente positiva nos grânulos citoplasmáticos. O teste para carboidratos foi fortemente positivo em todo o citoplasma, enquanto para os lipídeos observou-se reação positiva nas vesículas citoplasmáticas. Em relação às células de urato, estas apresentaram reação positiva para proteínas básicas no núcleo e citoplasma, por entre as vesículas. Essas células não apresentaram reação para o teste de PAS e Sudan Black B. Quanto aos enócitos, estes apresentaram citoplasma fracamente positivo ao PAS e fortemente positivo ao Sudan Black B e para o azul de bromofenol.

Macromolecular array patterns of silk gland secretion in social Hymenoptera larvae

The cocoon, produced by most holometabolous insects, is built with silk that is usually produced by the larval salivary gland. Although this silk has been widely studied in the Lepidoptera, its composition and macromolecular arrangement remains unknown in the Hymenoptera. The macromolecular array patterns of the silk in the larval salivary gland of some meliponids, wasps, and ants were analyzed with polarized-light microscopy, and they were compared with those of Bombyx mori (Lepidoptera). There is a birefringent secretion in the glandular lumen of all larvae, due to filamentous structural proteins that display anisotropy. The silk in the distal, middle and proximal regions of the secretory portion of Formicidae and Vespidae glands presented a lattice optical pattern. We found a different pattern in the middle secretory portion of the Meliponini, with a zigzag rather than a lattice pattern. This indicates that the biopolymer fibers begin their macromolecular reorganization at this glandular region, different from the Formicidae and the Vespidae, in which the zigzag optical pattern was only found at the lateral duct. Probably, the mechanism of silk production in the Hymenoptera is a characteristic inherited from a common ancestor of Vespoidea and Sphecoidea; the alterations in the pattern observed in the Meliponini could be a derived characteristic in the Hymenoptera. We found no similarity in the macromolecular reorganization patterns of the silk between the Hymenoptera species and the silkworm.