Physical mapping of the Nile tilapia (Oreochromis niloticus) genome by fluorescent in situ hybridization of repetitive DNAs to metaphase chromosomes - a review

The Nile tilapia (Oreochromis niloticus) has received increasing scientific interest over the past few decades for two reasons: first, tilapia is an enormously important species in aquaculture worldwide, especially in regions where there is a chronic shortage of animal protein; and second, this teleost fish belongs to the fascinating group of cichlid fishes that have undergone a rapid and extensive radiation of much interest to evolutionary biologists. Currently, studies based on physical and genetic mapping of the Nile tilapia genome offer the best opportunities for applying genomics to such diverse questions and issues as phylogeography, isolation of quantitative trait loci involved in behaviour, morphology, and disease, and overall improvement of aquacultural stocks. In this review, we have integrated molecular cytogenetic data for the Nile tilapia describing the chromosomal location of the repetitive DNA sequences, satellite DNAs, telomeres, 45S and 5S rDNAs, and the short and long interspersed nucleotide elements [short interspersed nuclear elements (SINEs) and long interspersed nuclear elements (LINEs)], and provide the beginnings of a physical genome map for this important teleost fish. (C) 2004 Elsevier B.V. All rights reserved.

Comparative cytogenetics of cichlid fishes through genomic in-situ hybridization (GISH) with emphasis on Oreochromis niloticus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Physical chromosome mapping of repetitive DNA sequences in Nile tilapia Oreochromis niloticus: Evidences for a differential distribution of repetitive elements in the sex chromosomes

Repetitive DNAs have been extensively applied as physical chromosome markers on comparative studies, identification of chromosome rearrangements and sex chromosomes, chromosome evolution analysis, and applied genetics. Here we report the characterization of repetitive DNA sequences from the Nile tilapia (Oreochromis niloticus) genome by construction and screening of plasmid library enriched with repetitive DNAs, analysis of a BAC-based physical map, and hybridization to chromosomes. The physical mapping of BACs enriched with repetitive sequences and C(o)t-1 DNA (DNA enriched for highly and moderately repetitive DNA sequences) to chromosomes using FISH showed a predominant distribution of repetitive elements in the centromeric and telomeric regions and along the entire length of the largest chromosome pair (X and Y sex chromosomes) of the species. The distribution of repetitive DNAs differed significantly between the p arm of X and Y chromosomes. These findings suggest that repetitive DNAs have had an important role in the differentiation of sex chromosomes. (c) 2007 Elsevier Ltd. All rights reserved.

Analytical approach to the metallomic of Nile tilapia (Oreochromis niloticus) liver tissue by SRXRF and FAAS after 2D-PAGE separation: Preliminary results

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Application of the comet assay in erythrocytes of Oreochromis niloticus (Pisces): a methodological comparison

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Induction of micronucleus and nuclear alterations in fish (Oreochromis niloticus) by a petroleum

In this study, micronucleus and nuclear alterations tests were performed on erythrocytes of Oreochromis niloticus (Perciformes, Cichlidae) in order to evaluate the water quality of the Atibaia river, in an area that receives effluents discharge of a petroleum refinery and also to evaluate the effectiveness of the treatments used by the refinery. Water samples were collected in five different sites related with a refinery from São Paulo State, Brazil. For the micronucleus and nuclear alterations tests, O. niloticus specimens were exposed for 72 h to the water samples and in pure ground water (negative control). The results herein obtained indicated that the treatments used by the refinery diminished the cytogenetic damage; however they were not fully effective, since the final mill has induced damages in the genetic material of the test organism. (C) 2008 Elsevier B.V. All rights reserved.

Genomic organization and comparative chromosome mapping of the U1 snRNA gene in cichlid fish, with an emphasis in Oreochromis niloticus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genotoxicity and mutagenicity of water contaminated with tannery effluents, as evaluated by the micronucleus test and comet assay using the fish Oreochromis niloticus and chromosome aberrations in onion root-tips

Cytotoxicity of metals is important because some metals are potential mutagens able to induce tumors in humans and experimental animals. Chromium can damage DNA in several ways, including DNA double strand breaks (DSBs) which generate chromosomal aberrations, micronucleus formation, sister chromatid exchange, formation of DNA adducts and alterations in DNA replication and transcription. In our study, water samples from three sites in the Córrego dos Bagres stream in the Franca municipality of the Brazilian state of São Paulo were subjected to the comet assay and micronucleus test using erythrocytes from the fish Oreochromis niloticus. Nuclear abnormalities of the erythrocytes included blebbed, notched and lobed nuclei, probably due to genotoxic chromium compounds. The greatest comet assay damage occurred with water from a chromium-containing tannery effluent discharge site, supporting the hypothesis that chromium residues can be genotoxic. The mutagenicity of the water samples was assessed using the onion root-tip cell assay, the most frequent chromosomal abnormalities observed being: c-metaphases, stick chromosome, chromosome breaks and losses, bridged anaphases, multipolar anaphases, and micronucleated and binucleated cells. Onion root-tip cell mutagenicity was highest for water samples containing the highest levels of chromium.

Partial molecular characterization of the Nile tilapia (Oreochromis niloticus) alpha-cardiac muscle actin gene and its relationship to actin isoforms of other fish species

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Respostas da Tilápia do Nilo (Oreochromis niloticus) à atratividade e palatabilidade de ingredientes utilizados na alimentação de peixes

Avaliaram-se as respostas da tilápia do Nilo à atratividade e palatabilidade de 14 ingredientes. O método utilizado foi de dupla escolha, comparando-se cada ingrediente peletizado à uma ração controle. Foram empregados quatro aquários (750 litros) contendo, cada um, três alevinos e dois comedouros instalados nos cantos direito e esquerdo, sendo registrados tempo decorrido entre colocação do ingrediente e chegada dos animais aos comedouros, freqüência de visitas aos comedouros, número de grânulos ingeridos e freqüência de ejeções. Os tratamentos foram comparados pela prova não-paramétrica de Kruskal-Wallis e a comparação dos ingredientes para o conjunto de variáveis, por intermédio da Análise de Agrupamento e Análise de Componentes Principais. Os ingredientes foram assim classificados: a) baixa atrato-palatabilidade = farelos de trigo, soja e algodão, farinha e raspa de mandioca, farinha de girassol e fubá de milho, b) média atrato-palatabilidade = levedura de cana-de-açúcar e glúten de milho e c) alta atrato-palatabilidade = ovo integral liofilizado, farinhas de crisálidas, peixes, carne e camarão. Ingestão de grânulos (manhã e tarde) e freqüência de visitas aos comedouros (tarde) foram consideradas as variáveis mais discriminatórias e freqüência de ejeção de grânulos e tempo gasto para aproximação dos comedouros (manhã e tarde), as menos discriminatórias. As respostas comportamentais dos peixes variaram de acordo com o ingrediente apresentado. A avaliação do grau de atrato-palatabilidade deve ser realizada considerando-se uma combinação de parâmetros.

Digestibilidade aparente de macrófitas aquáticas pela tilápia-do-nilo (Oreochromis niloticus) e qualidade da água em relação às concentrações de nutrientes

Objetivou-se com este estudo determinar os coeficientes de digestibilidade aparente (CDA) da proteína bruta e dos aminoácidos de duas espécies de macrófitas aquáticas flutuantes (Eichhornia crassipes e Pistia stratiotes) pela tilápia-do-nilo (Oreochromis niloticus) e verificar a qualidade da água dos aquários de digestibilidade em relação às concentrações de nitrogênio e fósforo. Foram elaboradas três rações, marcadas com 0,10% de óxido de cromo-III, sendo uma ração-referência (purificada) e as demais contendo 30% de cada uma das macrófitas aquáticas. As tilápias-do-nilo (58,8 + 18,5 g) foram alimentadas até a saciedade aparente e a coleta de fezes foi feita pelo sistema Guelph modificado. Os CDA médios da proteína e dos aminoácidos foram, respectivamente, 93,17 e 93,32% para a ração-referência; 59,23 e 60,35% para E. crassipes; e 52,24 e 57,40% para P. stratiotes. Não foram constatadas diferenças significativas entre os valores de CDA da proteína e dos aminoácidos dos ingredientes vegetais. Os resultados obtidos demonstraram reduzida eficiência da tilápia-do-nilo em assimilar a maioria dos aminoácidos de E. crassipes e P. stratiotes. As excretas das tilápias-do-nilo contribuíram para o aumento das concentrações de nitrogênio e fósforo na água dos aquários, independentemente da ração fornecida.

Physiological responses of Nile tilapia, Oreochromis niloticus, fed vitamin C- and lipid-supplemented diets and submitted to low-temperature stress

Water temperature alterations can determine harmful physiological modifications in fish, which should be prepared to cope with this, and nutrition strategies seem to be essential. This study evaluated the effects of different levels of vitamin C and lipids on physiological responses of Nile tilapia, Oreochromis niloticus, submitted to temperature stress. There were two phases: Phase I - preparing fish to store vitamin C and lipid at appropriate temperature, and Phase II - evaluating the contributions these reserves make to fish physiology under low-temperature stress. The experiment used a 3 x 2 factorial design with three vitamin C levels (300, 600, and 1200 mg/kg diet) and two lipid levels (8.0 and 12.0%), plus absence of nutrient test and a diet of 6.0% lipids and 125.0 mg/kg vitamin C. In Phase I, 192 fish were kept at 26.0 +/- 1.0 C for 112 d, and in Phase II, 48 fish were kept at 18.0 +/- 0.5 C for 32 d and at 15.0 +/- 0.5 C for 11 d. Fish fed C0L0 diet showed lower erythrocytes values in both phases; higher vitamin C supplement determined higher red blood cell (RBC) number and higher hematocrit (Htc) (Phase II); Htc was significantly lower in Phase II; after temperature stress, fish fed C0L0 diet had higher mean corpuscular volume, lower hemoglobin corpuscular concentration, and significantly lower vitamin C concentration in the liver; and higher supplementation determined a higher concentration in the liver (Phases I and II). Higher plasmatic cortisol concentration was seen in fish fed C0L0 diet. In conclusion, our results show that the absence of vitamin C in diets impairs RBC formation and does not enable fish to cope with stress; excess vitamin C is efficient in mitigating stress and 600 mg/kg diet is economic and physiologically sufficient to prepare fish for coping with low-temperature stress. Lipid supplementation does not determine alterations in stress biochemical parameters.

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.