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.

Histoenzymological and ultrastructural changes in lateral muscle fibers of Oreochromis niloticus (Teleostei: Cichlidae) after local injection of veratrine

The effects of veratrine have been investigated in mammalian, amphibian, and crustacean muscle, but not in fish. In this work, the action of veratrine was studied in the lateral muscle of the freshwater teleost Oreochromis niloticus after intramuscular injection. Histoenzymological typing and electron microscopy of muscle fibers before and 15, 30, and 60 min after veratrine injection (10 ng/kg fish) were used to indirectly assess the morphological changes and the oxidative and m-ATPase activities. In some cases, muscles were pretreated with tetrodotoxin to determine whether the ultrastructural changes were the result of Na+ channel activation by veratrine. Veratrine altered the metabolism of fibers mainly after 30 min. Oxidative fibers showed decreased NADH-TR activity, whereas that of glycolytic and oxidative-glycolytic type fibers increased. There was no change in the m-ATPase activity of the three fiber types, except at 60 min postveratrine, when a novel fiber type, which showed no reversal after acidic and alkaline preincubations, appeared. Ultrastructural damage involved sarcomeres, myofibrils, and mitochondria, but the T-tubules remained intact. Pretreatment with tetrodotoxin (1 ng/ml) prevented the ultrastructural changes caused by veratrine. These results show that in fish skeletal muscle veratrine produces some effects that are not seen in mammalian muscle.

Molecular and cytogenetic analysis of the telomeric (TTAGGG)n repetitive sequences in the Nile tilapia, Oreochromis niloticus (Teleostei: Cichlidae)

The majority of chromosomes in Oreochromis niloticus, as with most fish karyotyped to date, cannot be individually identified owing to their small size. As a first step in establishing a physical map for this important aquaculture species of tilapia we have analyzed the location of the vertebrate telomeric repeat sequence, (TTAGGG)n, in O. niloticus. Southern blot hybridization analysis and a Bal31 sensitivity assay confirm that the vertebrate telomeric repeat is indeed present at O. niloticus chromosomal ends with repeat tracts extending for 4-10 kb on chromosomal ends in erythrocytes. Fluorescent in situ hybridization revealed that (TTAGGG)n is found not only at telomeres, but also at two interstitial loci on chromosome 1. These data support the hypothesis that chromosome 1, which is significantly larger than all the other chromosomes in the karyotype, was produced by the fusion of three chromosomes and explain the overall reduction of chromosomal number from the ancestral teleost karyotype of 2n=48 to 2n=44 observed in tilapia.

Production of Nile tilapia Oreochromis niloticus and freshwater prawn Macrobrachium rosenbergii stocked at different densities in polyculture systems in Brazil

The effect of stocking prawns Macrobrachium rosenbergii at increasing densities in ponds with Nile tilapia Oreochromis niloticus reared at low density was evaluated. Twelve 0.01-ha earthen ponds were stocked with 1 tilapia/m2 and 0, 2, 4, or 6 post-larvae prawn/m2. Three replicates were randomly assigned to each prawn density. Postlarval prawns were stocked a week prior to tilapia juveniles and both were harvested 175 d after the beginning of the experiment. Tilapia final average weight, survival, production, and food conversion rates did not differ significantly among treatments (P > 0.05); the averages were 531 g. 67%, 3,673 kg/ha, and 1.91, respectively. Prawn survival rates did not differ for the three stocking densities (mean 90%). However, final weight and production were significantly different (P < 0.05) as follows: 34.0, 23.0, and 14.7 g and 639, 909, and 818 kg/ha, respectively for 2, 4, and 6 prawns/m2 densities. Stocking densities up to 6 prawn/m2 did not affect tilapia production and required neither additional feeding nor significant changes in management. The polyculture system allowed an increase in total production with the same amount of supplied feed, thus improving the system sustainability.