Marcelo Hideo Okamoto

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.

NPY and sbGnRH gene expression in juvenile and adult male Brazilian flounder Paralichthys orbignyanus

The objective of this study was to evaluate neuropeptide Y (NPY) and sea bream gonadotropin-release hormone (sbGnRH) gene expression in juvenile and adult males of Brazilian flounder. Hypothalamuses from fish were sampled for total RNA extraction. After cDNA synthesis, real-time PCR was used to measure gene expression. NPY showed approximately 2-fold increases in their mRNA levels while sbGnRH showed 3-fold increases in adult fish. These results suggest that these peptides could be involved on hypothalamic regulation of Brazilian flounder sexual maturation.

Oxidative stress parameters in juvenile Brazilian flounder Paralichthys orbignyanus (Valenciennes, 1839) (Pleuronectiformes: Paralichthyidae) exposed to cold and heat shocks

The aim of this study was to determine oxidative stress parameters in the liver and gill of Brazilian flounder juveniles (307.0 ± 16.0 g and 30.0 ± 4.0 cm) submitted to different water temperature (17.1, 23.0 and 28.8oC) for 72 h and maintained at salinity 25‰. After the acclimation of 7 days, in 23oC, fish were transferred to 200 L tanks containing seawater (salinity 25‰) at 28.8oC (heat shock), 17.1oC (cold shock) or 23.0oC (control), five replicates (five fish tank-1). The sampled collection occurred in 0 (pre-challenge), 3, 24, 48 and 72 h after temperature shock. Flounder exposed to 17.1oC and 28.8oC showed significantly higher TBARS levels and GST activity in the liver post-exposition (PE) in relation to the control (23oC). CAT activity in liver present a significantly increase at 17.1oC, in first 48 h, and subsequently decrease in 72 h PE in relation to 28.8oC. The gills of flounder showed significantly higher TBARS levels, GST and CAT activity when submitted at 17.1 and 28.8oC in relation to 23.0oC. There were observed changes in lipid peroxidation levels (LPO), CAT and GST activities in the liver and gill of Brazilian flounder in response to reactive oxygen species (ROS) produced by thermal shocks.

Taxa e frequência alimentar no crescimento de juvenis de pampo

The objective of this work was to evaluate the ideal feeding rate and frequency for juvenile pompano (Trachinotus marginatus). Two experiments were carried out in a completely randomized design, with three replicates each. In experiment I, 25 fish (4.8±0.6 g and 6.48±0.01 cm) were stocked in 15 tanks (50 L) during 21 days and fed 4, 8, 12, 16, and 20% body weight per day. In experiment II, 20 fish (4.1±0.1 g and 6.6±0.1 cm) were stocked in 15 tanks (40 L) during 28 days and fed 2, 6, 8, and 10 times a day. The tested feeding rates and frequencies did not influence survival. Final weight and length in experiment I were significantly lower in fish fed 4% body weight per day, whereas in experiment II only weight was significantly lower in fish fed 2 and 6 times a day. At the end of both experiments, apparent feed conversion showed significant difference, with the worst value observed for fish fed 20% body weight per day in experiment I and 2 times a day in experiment II. Juvenile pompano show better growth performance when fed 8% body weight per day and 8 times a day.

Clonagem e avaliação da expressão gênica do sbGnRH em machos juvenis e adultos de linguado, Paralichthys orbignyanus

The objectives of this study were to clone sbGnRH cDNA, evaluate the mRNA levels in different tissues of flounder, and also evaluate brain sbGnRH expression in juvenile and adult males. Using RT-PCR the cloning of a 297 nucleotides coding region of sbGnRH from Brazilian flounder brain was demonstrated for the first time. Expression of sbGnRH was detected in several peripheral tissues. Brain gene expression in the adult flounder was higher than those found in juvenile. These results suggest that sbGnRH is involved on the Brazilian flounder puberty.

Sclerosing hemangioma mandibular in Paralichthys orbignyanus: case report

In this study, we described the first case of sclerosing haemangioma in an adult Brazilian flounder Paralichthys orbignyanus. Produced by artificial reproduction, the fish was approximately 10 years old and was maintained at a breeding stock. When removed from the tank, mandibular lesion with excoriations and hemorrhagic foci were noted. Tumor samples were collected from the mandible for histopathological analysis. Proliferation of numerous blood vessels surrounded by dense connective stroma was observed microscopically. The etiology of this neoplasia is unknown, but the fact that the specimen remained in captivity for many years, may have contributed to the appearance of this type of lesion, due to sporadic mechanical shocks to the tank wall.