Environmental cadmium exposure and metabolic responses of the Nile tilapia, Oreochromis niloticus

The contamination of water by metal compounds is a worldwide environmental problem. This study was undertaken to evaluate the impact of short-term cadmium exposure on metabolic patterns of the freshwater fish Oreochromis niloticus. The fish were exposed to 320, 640, 1280 and 2560 mug/l sublethal concentrations of Cd++ (CdCl2) in water for 7 days. The specific activities of the enzymes phosphofructo kinase (PFK-E.C.2.7.1.11.), lactate dehydrogenase (LDH-E.C.1.1.1.27.) and creatine kinase (CK-E.C.2.7.3.2.) were decreased in white muscle after cadmium treatments, indicating decreases in the capacity of glycolysis in this tissue. Cadmium exposure induced increased glucose concentration in white muscle of fish. on the other hand, cadmium exposure at sublethal concentrations increased phosphofructo kinase and LDH in red muscle of fish. Cadmium significantly decreased total protein concentrations in liver and white muscle regardless of tissue glycogen levels. The data suggest that cadmium acts as a stressor, leading to metabolic alterations similar to those observed in starvation. (C) 2001 Elsevier B.V. Ltd. All rights reserved.

Relação Cálcio/Fósforo Disponível em Rações para Tilápia do Nilo (Oreochromis niloticus)

The objective of this work was to evaluate the effect of Ca: available P (P-a) ratio in the diets on performance and bone mineral depositions of juvenile Nile tilapia (2.5 +/- 0.5g). Seven isoproteic and isoenergetic diets (28% CP and 3100 Kcal of DE/Kg of diet), with similar crude fiber and sulfur aminoacids content, were used. The diets had 0.50 and 0.25; 0.50 and 0.50; 0.50 and 0.75; 0.80 and 0.40; 0.80 and 0.80; and 0.80 and 1.20% of Ca and P-a which corresponded to Ca/P-a ratios of 2:1;1:1 and 1:1.5, respectively. An additional control diet, with low levels of Ca and Pa (0.02 and 0.12%, respectively) was used. A completely randomized design with a 2'3 factorial arrangement (Ca level and Ca:P-a ratio) on treatments, plus the additional control diet and three replications was used. Five fishes per aquaria were randomly allotted to 21 fiberglass aquaria (80L). The temperature of the water was maintained close to 26degreesC, and the oxygen, pH and ammonia levels of the water were monitored. The results showed that it was necessary a minimum dietary level of 0.25% of P-a for a satisfactory bone mineral depositions, and that the best performance results were achieved using diets with Ca: P-a ratios between 1:1 and 1:1.5.

Sex determination studies in two species of teleost fish, Oreochromis niloticus and Leporinus elongatus

Genetic analyses of sex determination have identified sex chromosomes in many teleost fish species. However, there are several cases for which sex ratios do not fit perfectly with the expectations of heterogametic systems, suggesting the influence of either minor sex determining genes or environmental influences on the process of sex differentiation. The frequent absence of sex chromosome markers makes the identification of minor sex-determining genes very difficult. It is easier to test first the hypothesis of environmental sex determination (ESD) by studying the temperature effect, since temperature-dependent sex determination has been demonstrated to occur in several vertebrate groups including 1 fish species. To contribute to a better understanding of fish sex determination, we have tested the effects of high temperatures on sex ratios of Oreochromis niloticus, and have attempted to isolate sex chromosome molecular markers in Leporinus elongatus. Treatments of O. niloticus fry at 36 degrees C applied for 10 days and more, and starting 1 week after fertilization markedly increased the proportion of males, and progeny-testing these males confirmed that some of them are sex-reversed genetic females. Two non-coding sequences of L. elongatus Z and W chromosomes were cloned by genomic subtraction. They cross-hybridized with the genome of a close species without providing sex-specific patterns. A collection of L. elongates individuals was subjected to gonadal and chromosomal sexing, and DNA hybridization with both sequences. These analyses revealed 3 individuals having atypical W chromosomes. Interestingly, 2 of these were males having a ZW karyotype. We assume that these atypical sex chromosome arise by exchanges between Z and W chromosomes, and that a transition between female and male heterogamety is underway in this species.

Use of ideal protein concept for precision formulation of amino acid levels in fish-meal-free diets for juvenile Nile tilapia (Oreochromis niloticus L.)

This study was undertaken in a closed system with Nile tilapia (Oreochromis niloticus) to examine the effects of total replacement of fish meal (FM) by soybean meal. Nile tilapia fingerlings with an average weight of 5.34+/-0.08 g were hand-fed one of the five isoenergetic (approximate to13.5 MJ digestible energy kg(-1)) and isoproteic (approximate to31% of digestible protein) experimental diets to satiation, six times a day during 85 days in eight replicate fibreglass tanks (six fish per tank). The control diet containing FM was substituted by soybean meal, with and without essential amino acids (lysine, methionine and threonine) or dicalcium phosphate supplementation. The supplemental amino acids were added at levels to simulate the reference amino acid profile of Nile tilapia carcass protein, based on the ideal protein concept. The results showed that soybean meal diet supplemented only with dicalcium phosphate was inferior to the control diet with FM and soybean meal diets supplemented with dicalcium phosphate and essential amino acids. Multiple essential amino acids and dicalcium phosphate incorporation in soybean meal diets was associated with performance, whole-body composition and carcass yield equal to that of the fish fed with the control diet containing FM. These data suggest that a diet with all plant protein source, supplemented with essential amino acids, based on tissue amino acid profile, can totally replace FM in a diet for Nile tilapia, without adverse effects on the growth performance, carcass yield and composition.

SYNAPTONEMAL COMPLEX-ANALYSIS IN SPERMATOCYTES OF TILAPIA, OREOCHROMIS-NILOTICUS (PISCES, CICHLIDAE)

Some adaptations of the synaptonemal complex (SC) whole-mounting technique first used in plants permitted its application to meiotic studies in tilapia, Oreochromis niloticus. Direct observation of the chromosome pairing process and bivalent structure during the meiotic prophase of this fish species by light and electron microscopy permitted the analysis of SCs in autosomes and the possible identification of sex chromosomes. The analysis of SCs in spermatocytes of 0. niloticus revealed that all 22 bivalent chromosomes completely paired, except for the occurrence of a size heteromorphism in the terminal region of the largest bivalent associated with the presence of an incompletely paired segment during the synapsis process, which may be the cytological visualization of an XX/XY sex chromosome system in this species.

A LINE2 repetitive DNA sequence from the cichlid fish, Oreochromis niloticus: sequence analysis and chromosomal distribution

We report the cloning and characterization of a long interspersed nucleotide element (LINE) fi-om a cichlid fish, Oreochromis niloticus, and show the distribution of this element, called CiLINE2 for cichlid LINE2, in the chromosomes of this species. The identification of an open reading frame in CiLINE2 with amino acid sequence similarity to reverse transcriptases encoded by LINE-like elements in Caenorhabditis elegans, Platemys spixii, Schistosoma mansoni, Gallus gallus (CRI), Drosophila melanogaster (I factor), and Homo sapiens (LINE2), as well as the structure of the element, suggest it is a member of this family of non-long terminal repeat-containing retrotransposons. Search of a DNA sequence database identified sequences similar to CiLINE2 in four other fish species (Haplotaxodon microlepis, Oreochromis mossambicus, Pseudotropheus zebra, and Fugu rubripes). Southern blot hybridization experiments revealed the presence of sequences similar to CiLINE2 in all Tilapiini species analyzed from the genera Oreochromis, Tilapia, and Sarotherodon, and gave an estimated copy number of about 5500 for the haploid genome of O. niloticus. Fluorescent in situ hybridization showed that CiLINE2 sequences were organized in small clusters dispersed over all chromosomes of O. niloticus, with a higher concentration near chromosome ends. Furthermore the long arm of chromosome 1 was strikingly enriched with this sequence. The distribution of LINE2-related elements might underlie the difference in chromosome banding patterns observed between cold-blooded vertebrates and mammals.

Digestibilidade Aparente e Efeito Macro-Microscópico em Tilápia do Nilo (Oreochromis niloticus) Arraçoadas com Torta de Dendê

To evaluate the nutritional value of palm kernel meal on the performance Nile tilapia (Oreochromis niloticus), five isonitrogenous (30% crude protein), isoenergetic (2800 kcal/kg of digestible energy), and isofibrous (10% crude fiber) diets, with increasing levels of palm kernel meal (0, 7, 14, 21, 28 and 35%). were ad libitum fed to fingerlings with initial body weight of 1.52g +/- 0.04, housed per 120 days in 35 aquarium of 60 liters with fingerlings each. The environmental culture condition were monitored during all experimental period. Statistical analyses of recorded data were performed through polynomial regression models. To determine the apparent digestibility, macroscopic analysis of viscera and microscopic analysis of intestine epithelium were performed. The obtained results indicated that the inclusion of palm kernel meal up to 35% Nile tilapia diets did not affects the apparent digestibility and did not induce any pathological effects on viscera and intestinal epithelium.

Diversidade genética de quatro linhagens de Oreochromis niloticus utilizando o marcador RAPD

The aim of this study was to analyze the genetic diversity of four Nile Tilapia (Oreochromis niloticus) strains using the RAPD marker. Fin samples of GIFT (G), Chitralada (C), Supreme (S) and Bouake (B) juvenile stocks have been collected. The 11 primers used yielded 81 fragments of which 41.98% were polymorphic. The percentage of polymorphic loci (G: 18.52%; C: 19.75%; S: 20.99% and B: 24.79%) showed that there was a genetic differentiation among the strains, showing the G(st) values a high (BxG: 0.231; BxC: 0.224; GxC: 0.194 and SxC: 0.208) and elevated (BxS: 0.315 and GxS: 0.270) differentiation. The highest gene flow (N(m)) was among the GxC (2.082) strains. The distance and genetic identity values (0.044 and 0.957 respectively) and the dendrogram indicate that the GxC is the most genetically similar strains. The genetic similarity was high among of the strains (G: 0,932; C: 0,903; S: 0,891 and B: 0.900). These results will enable a correct reproductive and genetic strains management.

Effects of environmental colour on growth of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758), maintained individually or in groups

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mutagenic and genotoxic effects of the Atrazine herbicide in Oreochromis niloticus (Perciformes, Cichlidae) detected by the micronuclei test and the comet assay

Atrazine is the triazinic herbicide most found in the rural aquatic environments due to its extensive use and its stability in such places. The mutagenicity and the genotoxicity of different concentrations of the Atrazine herbicide were determinated by the micronucleus test and the comet assay, using Oreochromis niloticus as test-system. The tested concentrations of Atrazine herbicide were 6.25, 12.5 and 25 mu g/L, both for the micronuclei test and for the comet assay. The results showed a significant rate of micronuclei and nuclear abnormalities for all the tested concentrations of Atrazine herbicide. For the comet assay, we also observed results significantly different from the control in 6.25, 12.5 and 25 mu g/L concentrations. Due to these results, we could infer that such herbicide may be dangerous to the lives of those organisms exposed to it. (c) 2007 Elsevier B.V. All rights reserved.

A New and Fast Technique to Generate Offspring after Germ Cells Transplantation in Adult Fish: The Nile Tilapia (Oreochromis niloticus) Model

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Selênio orgânico na alimentação de matrizes de tilápia-do-Nilo (Oreochromis niloticus)

Organic selenium in the diet of Nile tilapia (Oreochromis niloticus) broodstock. The aim of this study to test the effect of organic selenium supplementation in the diets of Nile tilapia broodstock. Growth, reproductive performance and performance of fingerlings were evaluated. Eighty adult fish were used, with average weight of 121.7 g and divided into groups of four animals (3 female and 1 male) stocked in 20 tanks with 1.5 cubic meters. The fish were fed five diets containing different levels of organic selenium (0.0, 0.25, 0.50, 0.75, and 1.0 mg kg-1 ration). In the experimental period, females from all tanks were captured and the eggs collected. The experimental design was entirely randomized, with five treatments and four replicates. To evaluate progeny performance, the fingerlings were shared in four aquariums and fed during 30 days with commercial diet (45% CP). The supplementation with organic selenium did not affect the productive performance as with the reproductive parameters of the broodstock (p > 0.05). on the other hand, the broodstock fed with 0.50 mg of organic selenium produced fingerlings that presented improved weight gain (p < 0.05) compared to fish fed the control diet.

Desempenho produtivo de alevinos de tilápia-do-nilo (Oreochromis niloticus) arraçoados com farinha de soja

This experiment was carried out in the Research Laboratory on Nutrition of Aquatic Organisms, Aquaculture Center UNESP, FMVZ - Botucatu, SP, Brazil. The study aimed to evaluate the effect of raw full-fat soybean meal on feeding in fingerlings of the Nile tilapia (Oreochromis niloticus). The suitable level of consumption of raw full-fat soybean meal (RSM) was determined by means of productive performance. The experimental design was a Multivariate Profile Analyses, with four treatments (four replicates each one). Each isoproteic (26% CP) and isoenergetic (3000 Kcal ME/ kg) diet received raw full-fat soybean meal at 6.0%, 12.0% and 18.0%. A treatment without soybean meal was used as control. Forty eight fingerlings with similar size (mean Wt ± SD: 6.47 ± 0.64 g; and mean Lt ± SD: 54.94 ± 7.79 mm) at the beginning of the experiments were used. No significant difference (p>0.05%) in weight gain among the four treatments was found. However, the treatments with 12.0 and 18.0% RSM showed a reduction in the coefficient of digestibility in these treatments. This shows a deleterious effect of the raw full-fat soybean meal at these considered levels (12.0% and 18.0% RSM).

Nucleotide sequence of 5S rDNA and localization of the ribosomal RNA genes to metaphase chromosomes of the Tilapiine cichlid fish, Oreochromis niloticus

In this study, we report the cloning and nucleotide sequence of PCR-generated 5S rDNA from the Tilapiine cichlid fish, Oreochromis niloticus. Two types of 5S rDNA were detected that differed by insertions and/or deletions and base substitutions within the non-transcribed spacer (NTS). Two 5S rDNA loci were observed by fluorescent in situ hybridization (FISH) in metaphase spreads of tilapia chromosomes. FISH using an 18S rDNA probe and silver nitrate sequential staining of 5S-FISH slides showed three 18S rDNA loci that are not syntenic to the 5S rDNA loci.