Aparecida Alves do Nascimento

Fish Gills Alterations as Potential Biomarkers of Environmental Quality in a Eutrophized Tropical River in South-Eastern Brazil

Gill anomalies in three common fish species of different taxonomic order, habitat dwelling and feeding habits (one Characiformes, Oligosarcus hepsetus; one Siluriformes, Hypostomus auroguttatus; and one Perciformes, Geophagus brasiliensis) from a eutrophized tropical river in south‐eastern in Brazil were compared. The aim of this study was to search for sentinel species that could be used as potential biomarkers of environmental quality. Most fish had gills with histological changes, namely epithelial lifting, interstitial oedema, leucocyte infiltration, hyperplasia of the epithelial cells, lamellar fusion, vasodilatation and necrosis. On the other hand, lamellar blood congestion and lamellar aneurysm, which are more serious and often irreversible changes, were recorded for the water column carnivorous O. hepsetus and, to a lesser extent, for the bottom‐dwelling detritivorous H. auroguttatus. A histopathological alteration index (HAI) based on the occurrence and severity of gills anomalies indicated that O. hepsetus (mean score = 11.4) had significantly higher values (Kruskall–Wallis H2,41 = 15.95, P = 0.0003) compared with G. brasiliensis (mean score = 7.0). Overall, the omnivorous G. brasiliensis had comparatively lesser occurrence of most gill anomalies compared with other two species, being less suitable as biomarker of environmental quality. In contrast, the water column–dweller O. hepsetus (water column) and the bottom‐dweller H. auroguttatus had gills most susceptible to changes, making them more suitable for using as histological biomarkers of the environmental quality in entrophized tropical rivers.

Histological and histochemical analysis of the gonadal development of males and females of Armases rubripes (Rathbun 1897) (Crustacea, Brachyura, Sesarmidae)

The objective of this study was to provide information on the histological characteristics of the gonads of male and female Armases rubripes crabs, and to try to establish a relationship between the microscopic and macroscopic stages previously identified. Thirty-six crabs were collected by hand between February 2003 and January 2004 in banks of Spartina alterniflora on Sahy Beach in Mangaratiba, Rio de Janeiro state, Brazil. The histological analysis of the ovaries of A. rubripes demonstrated a gradual process of development of the oocytes. According to their cellular characteristics, five types of cells were distinguished: oogonia, oocyte I, oocyte II, oocyte III and oocyte IV. The ovaries showed four stages during gonadal activity: stage I (rudimentary), stage II (developing or maturing), stage III (developed or mature) and stage IV (resting). The results of the histochemical analyses showed that the ovaries vary according to the gonad development stage. The histological aspect of one section of the male gonad was always the same in all of the seminiferous tubules, where the lumen of these tubules always contained spermatozoa and/or spermatids. It was not possible to characterize the three stages of gonad development in the males. This agrees with previous reports in the literature. However, in the females there was a relationship between the gonad stages distinguished macroscopically and the results obtained through the histological and histochemical analysis, due to the presence of different cell types, as well as the lysis process and reabsorption of the oocytes in spent females.

Correlação morfométrica do ovário de fêmeas bovinas em diferentes estádios reprodutivos

The aim of this study was to evaluate morphometric characteristics of bovine ovaries recovered on different reproductive stages. The ovaries used (N=200) were recovered at slaughterhouses and sent to the laboratory, were the size of the ovary, of follicles and of corpus luteum were measured. The pairs of ovaries were classified according to the reproductive stage, and to the estrous cycle or pregnancy stages of the animals from which they were recovered. There was a difference (P 0,05) on organ size between animals presenting estrous cycles or pregnancy, but pregnant cows presented larger corpora lutea and smaller diameter of the largest follicle (P<0,05). Evaluated parameters were affected by estrous cycle and pregnancy stages (P<0,05). There was a significant correlation between ovary and corpus luteum sizes in the animals presenting estrous cycles (R=0.69; P<0,001). In conclusion, ovarian morphologic characteristics changes according to the reproductive stages, could be used as parameters for clinic and functional evaluations of the organ.

Can fish gill anomalies be used to assess water quality in freshwater Neotropical systems

Gill anomalies in two fish species (Geophagus brasiliensis and Astyanax bimaculatus) were compared among three freshwater systems with different water quality: one eutrophic river, one eutrophic reservoir, and one oligotrophic reservoir. The raised hypotheses are that reservoirs with low water quality (eutrophic) have fish with more gills anomalies compared with reservoirs with high water quality (oligotrophic), and that the more stable environmental conditions of eutrophic rivers have fish with better healthy conditions than eutrophic reservoirs that have lesser stable environmental conditions. Gills of 36 adult individuals of G. brasiliensis and 23 of A. bimaculatus collected during the winter 2008 and winter 2009 were examined, and the proportions of occurrence of nine histological alterations were compared for the two species among the three systems using a binomial t test for independent samples. Histological changes in fish gills that are reversible and unspecific, such as epithelial lifting, interstitial edema, leukocyte infiltration, hyperplasia of the epithelial cells, lamellar fusion, and vasodilatation were common in both fish species in the three systems. However, lamellar aneurism, which is a more serious and often irreversible anomaly, and lamellar blood congestion occurred only in fish from the two reservoirs. Alternatively, necrosis occurred more in fish from the river. Fish gill anomalies in both species did not differ between the two reservoirs, despite having different water quality. We rejected the hypothesis that reservoirs with lower water quality have fish with more gill injuries compared with high water quality reservoirs. Moreover, the eutrophic river seems to affect differently the healthy condition of fish species, compared with the eutrophic reservoir.

Distributions of the endocrine cells in the gastrointestinal tract of nectarivorous and sanguivorous bats: A comparative immunocytochemical study

The present study was conducted to clarify the influence of feeding habits on regional distribution and relative frequency of endocrine cells secreting cholecystokinin (CCK), gastrin (GAS), serotonin (5-HT) and enteroglucagon (GLUC) in the nectarivorous Anoura geoffroyi and Glossophaga soricina and the sanguivorous Desmodus rotundus bats of the Phyllostomidae family, by specific immunohistochemical methods. The regional distribution and frequency of the different types of endocrine cells varied according to their location in the GIT. 5-HT immunoreactive cells (IR), detected throughout the GIT of three bats, were the most predominant gastrointestinal endocrine cells. GAS-IR cells in A. geoffroyi were found at the base of the pyloric gland, while in G. soricina they could also be observed in the middle to basal portions of the gland. GLUC-IR cells were located in the fundic region of A. geoffroyi, G. soricina and D. rotundus. These endocrine cells were more abundant in the sanguivorous bat. In nectarivorous bats were compared to sanguivorous bat, which differ in dietary habits, difference in the distribution and relative frequency of gut endocrine cells would be predicted. The absence of some, and decrease in frequency of other, gastrointestinal endocrine cells may reflect, in part, its interspecies differences or dietary habits.

Immunocytochemical study of gastrintestinal endocrine cells in insectivorous bats (Mammalia: Chiroptera)

The regional distribution and relative frequency of endocrine cells in the stomach and intestine of Phyllostomidae: Lonchorhina aurita and Molossidae: Molossus molossus bats were studied immunohistochemically. Three types of immunoreactive (IR) endocrine cells--to serotonin (5-HT), gastrin (GAS) and enteroglucagon (GLUC)--were found in the gastric mucosa and four types of IR cells were identified in the intestinal mucosa. This study showed an interespecfic difference in the regional distribution and relative frequency of endocrine cells in the Chiropteran alimentary tract.

Immunocytochemical study of the distribuition of endocrine cells in the pancreas of the Brazilian sparrow species Zonotrichia Capensis Subtorquata (Swaison, 1837)

In the present study, we investigated types of pancreatic endocrine cells and its respective peptides in the Brazilian sparrow species using immunocytochemistry. The use of polyclonal specific antisera for somatostatin, glucagon, avian pancreatic polypeptide (APP), YY polypeptide (PYY) and insulin, revealed a diversified distribution in the pancreas. All these types of immunoreactive cells were observed in the pancreas with different amounts. Insulin-Immunoreactive cells to (B cells) were most numerous, preferably occupying the central place in the pancreatic islets. Somatostatin, PPA, PYY and glucagon immunoreactive cells occurred in a lower frequency in the periphery of pancreatic islets.

Histological Description of Morphogenesis of the Gastroesophageal Mucosa of Gallus gallus domesticus (Linnaeus, 1758)

La morfogenesis de la mucosa gastroesofagica implica alteraciones morfologicas relacionadas con su estructura y la funcion de cada segmento. El presente estudio describe la histogenesis del epitelio secretor mucinoso y de las glandulas del esofago, molleja y proventriculo del pollo (G. gallus), ademas de identificar los cambios em el patron de secrecion de glicocosaminoglicanos (GAGs). Se utilizaron treinta y ocho embriones de pollo. El material recogido fue procesado de acuerdo a la rutina histologica y posteriormente las secciones se tinieron con hematoxilina-eosina para su analisis histologico, con tricromico de Gomori para identificar el tejido conectivo y las fibras de colageno y con PAS y AB pH 2,5 para el analisis GAGs neutro y acido. En una etapa avanzada de desarrollo de los embriones, se pudo obervar en la pared del esofago cuatro capas: mucosa, submucosa, muscular y serosa, mientras que el proventriculo y molleja se muestra constituida por tres capas: mucosa, muscular y serosa. En los tres segmentos de la superficie, el epitelio se identifico como mucinoso y en el esofago solo en la etapa inicial de desarrollo. Las glandulas del proventriculo se empiezan a formar en las primeras etapas de desarrollo, mientras que en las glandulas tubulares de la molleja comienzan su solo despues del dia 15. Las diferencias en la produccion de GAGs en estas regiones del tracto digestivo estan relacionadas con las etapas de desarrollo, las funciones y necesidades fisiologicas de cada segmento del cuerpo y se adapta gradualmente a la vida despues de la eclosion.

An immunocytochemical study of the endocrine cells in the stomach and duodenum of Zonotrichia capensis subtorquata (Passeriformes, Emberizidae)

The main purpose of this study was to evaluate the regional distribution pattern and relative frequency of some endocrine cells in the three portions of the gastrointestinal tract (GIT)--the proventriculus, gizzard and duodenum- of the rufous-collared sparrow (Zonotrichia capensis subtorquata), by immunohistochemical methods using six types of polyclonal antisera, specific for serotonin (5-HT), somatostatin (D cells), glucagon, motilin, polypeptide YY (PYY) and insulin. In the proventriculus, endocrine cells immunoreactive for all of these markers were observed. The somatostatin-immunoreactive cells were found with greater frequency, with the presence of cytoplasmic processes. In the gizzard, endocrine cells secreting somatostatin, 5-HT and PYY were detected, while those secreting glucagon and insulin were not. In the final part of the gizzard, endocrine cells secreting 5-HT were more frequent, and cells secreting somatostatin and insulin were not detected. All of the cell types studied were observed in the duodenum in different frequencies, except for cells immunoreactive for glucagon and insulin. The somatostatin-positive (D cells) were the most numerous, being more prevalent in the intestinal glands. The other endocrine cells were identified in smaller numbers, some of them located in the intestinal villi and Lieberkuhn glands. The finding of these cell types in the duodenum confirms their preferential location in the final portions of the principal segments of the digestive system and suggests control by feedback of its functions. In conclusion, some interesting distributional patterns of gastrointestinal endocrine cells were found in this species of sparrow.

Histochemical and immunohistochemical study on endocrine cells (5HT, GAS, and SST) of the gastrointestinal tract of a teleost, the characin Astyanax bimaculatus

Endocrine cells secrete hormones through the mucosa of the gastrointestinal tract (GIT) and act on the overall regulation of digestive processes such as nutrient absorption, gut motility and intestinal blood flow. This study aimed to determine regional distribution and frequency of endocrine cells secretory of serotonin (5-HT), somatostatin (SST) and gastrin (GAS) in the GIT of a small-bodied widespread characin Astyanax bimaculatus using histological, histochemical and immunohistochemical techniques. Fragments of the stomach and gut fixed for 8h in Bouin liquid were subjected to histological processing and immunohistochemical routine. For the histological analyses, the technique of staining with hematoxylin and eosin (HE) was used, whereas for the histochemical analyses Gomoris trichrome, periodic acid+Schiff (PAS) and Alcian blue pH 2.5 (AB) were used to further immunohistochemical processing. The stomach has a mucosa lined with a simple columnar epithelium with mucus-secreting cells; the glandular region (proximal and distal portions) has folds and pits, whereas the non-glandular region has pits only. The intestinal epithelium is simple with plain cylindrical grooved and goblet cells. The anterior region has thin folds with few goblet cells, and the posterior region with thick folds and many goblet cells. The regional distribution and frequency of endocrine cells varied across regions of the GIT with the stomach showing the highest amount of immunoreactive (IR) cells. Only the 5-HT was found in the stomach (epithelia and glands) and gut regions, with comparatively higher frequency in the stomach. SST-IR cells were found in the stomach (epithelia and gastric glands) with higher frequency in the glandular region, whereas GAS-IR were found in the gastric glands only. The stomach was the only organ to have all the three types of endocrine cells, indicating that this organ is the main site of digestion of food in this species.