Luís André Sampaio

Deleterious effects of water-soluble fraction of petroleum, diesel and gasoline on marine pejerrey Odontesthes argentinensis larvae.

Accidental discharges and oil spills are frequent around the world. Petroleum-derived hydrocarbons are considered one of the main pollutants of aquatic ecosystem. The importance of petroleum and refined fuels is notorious because todays society depends on them. Researches related to the toxic water-soluble fraction (WSF) of petroleum and derivatives to aquatic biota are scarce. For this reason, deleterious effects of WSF of Brazilian petroleum, automotive diesel and unleaded gasoline to marine pejerrey Odontesthes argentinensis larvae were studied employing toxicity tests and histopathological examination. Each WSF was generated in a laboratory by mixing four parts of seawater with one part of pollutant by approximately 22 h. Larvae were exposed during 96 h to different concentrations of WSF of petroleum, diesel, and gasoline, plus a control. After 96 h of exposure to the different WSFs, three larvae were sampled for histopathological studies. The median lethal concentration after 96 h (LC50) of exposure for WSF of petroleum was equal to 70.68%, it was significantly higher (P<0.05) than the values for WSF of diesel and gasoline, which were 13.46% and 5.48%, respectively. The histological examination of pejerrey larvae exposed to WSF of petroleum, diesel and gasoline after 96 h revealed a variety of lesions in the larvae. The gills, pseudobranchs and esophagus presented epithelial hyperplasia, and the liver presented dilatation of hepatic sinusoids, hepatocitomegaly, bi-nucleated and nuclear degeneration of hepatocytes, such as pyknotic nuclei. The acute toxicity of diesel and gasoline is at least fivefold higher than Brazilian petroleum. However, all toxicants induced histopathological abnormalities in pejerrey larvae. The results are of importance since much attention has been paid to large visible surfaces of petroleum spills instead of potential toxic effects of dissolved aromatic hydrocarbons, which are more available to marine biota.

Evaluation of DNase activity in seminal plasma and uptake of exogenous DNA by spermatozoa of the Brazilian flounder Paralichthys orbignyanus

Sperm mediated gene transfer (SMGT) has been successfully used in mammals, amphibians, birds, and some invertebrates. In fish, this methodology has failed or had poor efficiency for the production of transgenic specimens, presumably because the processes regulating the interaction between spermatozoa and exogenous DNA are not well understood. Therefore, the objective was to develop a SMGT protocol for the Brazilian flounder Paralichthys orbignyanus, with an emphasis on the role of seminal plasma DNase on exogenous DNA uptake by fish spermatozoa. In this study, there was strong DNase activity in the seminal plasma of P. orbignyanus; however, this DNase activity was decreased or eliminated by washing the spermatozoa with solutions containing EDTA (DNase activity was completely inhibited by 40 mM EDTA). Three washing solutions were tested, all of which maintained sperm quality. Moreover, it was determined that the no more than 50 ng of exogenous DNA/10(6) cells should be used for SMGT in fish. Finally, it was demonstrated that fish spermatozoa were capable of spontaneous uptake of exogenous DNA after elimination of DNase activity; this was confirmed by exogenous DNA amplification (PCR using sperm genomic DNA as a template) after DNase I treatment. We concluded that whereas DNase activity was an important obstacle for exogenous DNA uptake by fish spermatozoa; controlling this activity improved the efficiency of SMGT in fish.

Identification, tissue distribution and evaluation of brain neuropeptide Y gene expression in the Brazilian flounder Paralichthys orbignyanus.

Neuropeptide Y (NPY) is one of the most potent stimulants of food intake in vertebrates, mammals and fish. However, the present knowledge about feeding behaviour in fish is still limited and based on studies in a few species. The Brazilian flounder Paralichthys orbignyanus is being considered for aquaculture, and it is important to understand the mechanisms regulating feeding in order to improve its performance in captivity. The objectives of this study were to clone NPY cDNA, evaluate the mRNA levels in different tissues of flounder, and also evaluate brain NPY expression to associate food intake with NPY expression levels. A 597 bp NPY cDNA was cloned from Brazilian flounder brain. NPY expression was detected in all the peripheral tissues analysed. No significant differences were observed in brain NPY gene expression over 24 h after food intake at a temperature of 15 ± 3°C. No correlation was observed among plasma glucose, total protein, cholesterol, triglycerides and NPY expression levels during this 24 h period. On the other hand, mRNA levels were increased after two weeks of fasting at elevated temperatures. Our results suggest that NPY mRNA levels in Brazilian flounder are affected by temperature.

Benzocaína e eugenol como anestésicos para juvenis do pampo Trachinotus marginatus

The aim of this experiment was to evaluate the efficacy of benzocaine and eugenol as anesthetics for Trachinotus marginatus juveniles. Five concentrations of benzocaine (25, 50, 75, 100, and 150ppm) and eugenol (25, 37.5, 50, 62.5, and 75ppm) were tested. The trials were conducted in glass aquariums filled with 30l seawater (salinity 30‰ and temperature 19°C). The fish (51.4±13.5g) were individually exposed to different concentrations of each anesthetic (n=5 for each concentration). They were measured and weighed under anesthesia, and then returned to another aquarium without anesthetic for recovery. The results were evaluated using Analyses of Variance (One-Way - ANOVA) followed by the test of Duncan (95% confidence level) when necessary. Anesthesia time varied from approximately 11min (25ppm) to 2min (150ppm) for benzocaine, and from 10min (25ppm) to 2min (75ppm) for eugenol. Recovery time ranged from 3min (25ppm) to 14min (150ppm) for benzocaine and from 2min (25ppm) to 8min (75ppm) for eugenol. Benzocaine and eugenol, both at 50ppm, induce quick anesthesia and recovery (3mim for anesthesia and 5mim for recovery) for juvenile Trachinotus marginatus, and therefore are considered efficient anesthetics for this species. However, it is important to emphasize that the cost of benzocaine is 1/3 lower than eugenol to anesthetize juvenile pompano.

Growth of Juvenile Brazilian Flounder, Paralichthys orbignyanus, Cultured at Different Salinities

ABSTRACT The Brazilian flounder, Paralichthys orbignyanus, is found in coastal and estuarine waters of the Western South Atlantic Ocean. It is being considered for aquaculture due to its high market price and wide environmental tolerance. In this study, the hypothesis of enhanced growth of Brazilian flounder at a salinity equivalent to its isosmotic point was tested. Hatchery raised juveniles were cultured during 12 months at three salinities: 2,11, and 30 ppt in a semi-recirculation system (15-20% daily water renewal). Every 15 days, length and weight were measured. At the end of the study, flounders were sacrificed and their gonads sampled to determine sex and to ascertain gonad development. A greater proportion of males (approximately 90%) was observed at all salinities. All individuals were mature, but females contained no gametes beyond oocyte III. Growth was significantly higher at 30 ppt when compared to 2 ppt, but no differences were observed for flounders reared at 11 ppt. Thus the hypothesis that growth of Brazilian flounder could be enhanced in a isosmotic environment was refused. These results suggest that Brazilian flounder may be a suitable candidate for estuarine and coastal aquaculture, but additional information will be required before aquaculture facilities are developed in Brazil.

Effect of salinity on survival, growth and biochemical parameters in juvenile Lebranch mullet Mugil liza (Perciformes: Mugilidae)

Teleost fish growth may be improved under isosmotic condition. Growth and metabolic performance of juvenile Mugil liza (isosmotic point: 12‰) were evaluated after 40 days in different salinities (0, 6, 12 and 24‰). Tests were performed in quadruplicate (30 fish/tank; 0.48 ± 0.1 g body weight; 3.27 ± 0.1 cm total length) under controlled water temperature (28.2 ± 0.1oC) and oxygen content (>90% saturation). Fish were fed on artificial diet (50% crude protein) four times a day until apparent satiation. Results showed that salinity influenced juvenile mullet growth. Fish reared at salinity 24‰ grew better than those maintained in freshwater (salinity 0‰). Gill Na+,K+-ATPase activity and whole body oxygen consumption showed an U-shape-type response over the range of salinities tested, with the lower values being observed at the intermediate salinities. Although no significant difference was observed in liver glycogen content at different salinities, it tended to augment with increasing salinity. These findings indicate that energy demand for osmorregulation in juvenile M. liza can be minimized under isosmotic condition. However, the amount of energy spared is not enough to improve fish growth. Results also suggest that M. liza is able to alternate between different energy-rich substrates during acclimation to environmental salinity.

Growth of Critically Endangered annual fish Austrolebias wolterstorffi (Cyprinodontiformes: Rivulidae) at different temperatures

Given the importance of knowledge of the biology of endangered species for the planning of conservation and management efforts, the aim of this study was to investigate the influence of temperature on the growth of Austrolebias wolterstorffi (Ahl, 1924). To clarify the thermal influence on the growth of the species, temperatures of 16, 20, 24 , and 28°C were tested in triplicate. The present study showed that a water temperature of 28°C is detrimental to the growth of the species. Among the other tested temperatures, it was found that the optimum temperature for growth decreases as an individual ages and is slightly lower for females. It was demonstrated here that males reach a higher weight and length, and grow faster than females, reaching sexual maturity earlier. To optimize the growth of this species in captivity, the ideal temperature of the water during the initial life period is 24°C, until after puberty when the temperature should decrease to about 21°C.

Effect of temperature on growth of the threatened annual fish Austrolebias nigrofasciatus Costa & Cheffe 2001

This study evaluated the effect of temperature on growth of Austrolebias nigrofasciatus, an endemic and threatened annual killifish species of the Patos-Mirim lagoon system in Southern Brazil. In order to verify the effect of temperature on initial growth of A. nigrofasciatus, eggs stored in the laboratory were hatched and juveniles reared for eight weeks at 16 and 22 oC. The standard length of newly hatched fishes was 4.67 ± 0.25 mm and after eight weeks they reached 23.68 ± 3.73 and 22.68 ± 5.36 mm, respectively at 16 and 22 oC. However, initial growth of fish reared at 22 oC was faster and they reached sexual dimorphism at an earlier age compared to those reared at 16 oC. Final length of females reared at 22 oC was 23.00 ± 2.83 mm, they were significantly larger than those reared at 16 oC (17.91 ± 2.47 mm). Males were significantly larger than the females at 16 oC, but there was no difference for growth between sexes of fish reared at 22 oC. The sex ratios were 1:0.6 and 1:1.1 (M:F) at 16 oC and 22 oC, respectively, suggesting temperature determination of phenotypic sex. Considering the results, it appears that juveniles to be developed in captivity should be kept at 22 oC during the first six weeks of life, thus ensuring a higher growth rate until puberty.

Produção de juvenis do linguado Paralichthys orbignyanus: efeito da duração do período de co-alimentação durante o desmame

Weaning is a critical period for production of marine fish juveniles. The objective of this research was to evaluate the effect of co-feeding on survival, growth, and cost of production after the weaning of juveniles of Brazilian flounder Paralichthys orbignyanus. Juvenile flounder (32 days after hatching) were fed on HUFA enriched Artemia and subsequently three periods of co-feeding with dry diets (15, 20, and 25 days) were tested, a control group, where flounder were fed exclusively on Artemia was also followed until the end of the experiment. Juveniles fed on Artemia showed significantly higher (P 0.05) for juveniles fed on dry diets, but a higher survival rate was registered when the co-feeding period was longer. The results obtained demonstrate that flounder fed exclusively on Artemia have a better performance than those weaned into dry diets. However, unitary cost of production is reduced when juveniles are weaned 58 days after hatching.

Feeding Rate and Frequency Affect Growth of Juvenile Atlantic Spadefish

Abstract The Atlantic spadefish Chaetodipterus faber is an excellent candidate for aquaculture development, but success will depend on the identification of proper feeds and feeding regimens for this species. Accordingly, we evaluated the growth performance of juvenile Atlantic spadefish (3.60 ± 0.03 g [mean ± SE]) fed at rates of 3, 5, or 7% of body weight (BW) per day, either in a single feeding (1×) or in three equal feedings (3×). Weight gain, specific growth rate, feed conversion ratio, and feed intake were significantly affected by both feeding rate and frequency. Weight gain and the specific growth rate increased significantly with feeding rate, and growth was generally greater and more efficient in the 3× groups than in the 1× groups. Fish fed at higher feeding rates accumulated significantly more lipid within the body and had associated decreases in moisture, protein, and ash content, but carcass composition was unaffected by feeding frequency. We suggest that the growth of juvenile Atlantic spad...