Evaluation of heavy metal sediment toxicity in littoral ecosystems using juveniles of the fish Sparus aurata.

The toxicity of sediments from two littoral ecosystems of the Gulf of Cadiz was tested using juveniles of the fish Sparus aurata (seabream). Concentrations of total carbon and nitrogen, organic carbon, 14 heavy metals (Fe, Mn, Cu, Zn, Pb, Cd, Ag, Hg, As, Sn, V, Ni, Co, Cr), and the surfactant linear alkyl benzenesulfonate (LAS) in the sediments were measured. Chemical analysis was performed in the stations to determine the degree and nature of contamination. Four different endpoints were selected in the toxicity test: survival, superficial alteration, hematocrit analysis, and histological damage. After 14 days, survival, superficial alteration, and hematocrit analysis did not reveal effects of the different sediments tested. The histological and cellular damage revealed a more sensitive response to measured chemicals in sediments and they were found to be a powerful tool to evaluate sediment toxicity effects. Semiquantitative evaluation of the histological damage demonstrated correlation with sediment concentrations of some of the heavy metals (Cr, Cd, Pb, Ag, Cu) and the surfactant (LAS). Data derived from chemical concentrations and toxicity tests were assembled by multivariate statistical techniques (principal components analysis) to identify the ranges of chemical concentrations associated with an adverse effect. The results obtained, as suggested by site-specific sediment quality values, were the following: Cr>/=90.2; Cd>/=1.24; Pb>/=52.5; Ag>/=0.68; Cu>/=71.2; LAS>/=8.7 mg kg-1 of dry sediment. These results are mainly in concordance with studies performed in other areas of the world and therefore support wide application of the method.

Characterization of digestive enzyme activity during larval development of gilthead seabream (Sparus aurata).

The evolution of the digestive enzyme equipment in seabream from hatching to 30 days old larvae was studied; there was a progressive increase in the activity of protease, amylase and acid and alkaline phosphatase from day 15 onwards. The use of specific inhibitors, and SDS-PAGE provided evidence to suggest that most of the proteases belonged to the serine group. A high α-amylase activity was also denoted. Zymograms of larval extracts indicated that exogenous food has more a qualitative than a quantitative role in the secretion of digestive enzymes in this species.

A histochemical and immunohistochemical study of digestive enzymes and hormones during the larval development of the sea bream, Sparus aurata L.

The distribution of different hydrolytic enzymes and the localization of the hormones which regulate glucose metabolism during development of the digestive tract of the sea bream, Sparus aurata L., were studied. The yolk sac contains trypsin, glucose-6-phosphatase, ATPases and acid and alkaline phosphatase activities. Positive insulin, glucagon and somatostatin cells were observed in the pancreas and in the lumen of the intestinal tract during endogenous feeding. From hatching until 3 days later, the digestive tract of sea bream larvae shows no enzymatic activities. During exogenous feeding, the activities of the phosphatases and trypsin generally increase, as do the amounts of the hydrolytic enzymes and trypsin, as well as the pancreatic and intestinal hormones. The enzymatic activities gradually decrease from the anterior part towards the posterior part of the digestive tract.

New staining methods for semithin sections of tissues of Seriola dumerili embedded in methacrylate.

Semithin sections of different tissues of Seriola dumerili R. Embedded in glycol methacrylate, were stained with two staining methods normally used in conventional haematology and histology: Gutierrez's cytopanchrome and Gutierrez's polychrome V.O.F. The results observed in all tissues embedded in this resin and stained with V.O.F. are satisfactory and comparable, as to staining affinity, to those obtained on paraffin-embedded tissues. Gutierrez's trichrome stain appears to be a useful tool in the study of oogenesis for detecting changes in staining according to cellular structure and maturation stage. The results observed with cytopanchrome are similar to those obtained with toluidine blue in glycol methacrylate-embedded tissues. The use of these two staining techniques, requires no previous oxidation, alkali treatments, high temperatures or removal of the embedding medium. Furthermore, both dyes can be used at the same time: in this case, cytoplasm of previtellogenic oocytes showed a stronger affinity for cytopanchrome, and were more intensely stained by V.O.F. as the vitellogenesis of Seriola dumerili progressed. Both stain, and especially the cytopanchrome one, also are available for staining the liver, gills and testis.

Carbohydrate and protein histochemistry during oogenesis in Halobatrachus didactylus (Schneider, 1801) from the Bay of Cadiz (Spain).

Histological and histochemical characteristics were studied in Halobatrachus didactylus (Schneider, 1801) during oogenesis. Three phases could be differentiated: previtellogenesis (oogonia and basophilic oocytes), vitellogenesis (yolk synthesis) and maturation-spawning. Glycogen, glycoproteins and proteins rich in certain amino acids were present in the previtellogenic as well as in the vitellogenic cytoplasm oocytes. No acid mucosubstances were detected. Three types of yolk (vesicles, vacuoles and granules) contained different types of organic reserves; granules were essentially proteic whereas globules were lipidic. Carbohydrates and proteins were present in vesicles.

Comparative histopathological alterations in the digestive gland of marine bivalves exposed to Cu and Cd.

This study compares the histopathological alterations in the digestive gland cells of mussels, Mytilus galloprovincialis and clams, Ruditapes phillipinarum following exposure to copper and cadmium. The results show degenerative processes undergone in the digestive gland ranging from inflammatory responses to extreme vacuolation, particularly in Cd-exposed individuals. Unsaturated neutral lipids tend to accumulate in pathologically enlarged lysosomes of the homogeneous-type or heterogeneous-type depending of the species and of metal. Lipofucsins containing granules were mainly found in Cu-exposed mussels and Cd-exposed clams. No granules were detected in Cd-mussels. The comparison of the methods indicate that paraffin sections are also a suitable material for the localization of lipofucsins.