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.

Short interspersed repetitive elements (SINEs) from the cichlid fish, Oreochromis niloticus, and their chromosomal localization by fluorescent in situ hybridization

In the present study, we describe the cloning and characterization of a new SINE-like element from O. niloticus (ROn-2) and show the distribution of this SINE and a previously isolated SINE, ROn-1, in the chromosomes of O. niloticus. The ROn-2 element is 359 base pairs (bp) in length, contains short direct terminal repeats, a tRNA-related region similar to tRNA Val and tRNA Arg, a tRNA-unrelated region, and a poly-A tail. Analysis of the chromosomal distribution of ROn-1 and ROn-2 by fluorescent in situ hybridization showed that both SINE sequences are present in all chromosomes of tilapia, and organized in small clusters. The only exception was a large cluster of ROn-1 repeats found in the middle of the long arm of chromosome 1. In view of our data we discuss the hypothesis that the absence of large clusters of SINE sequences and the structural composition of these sequences may explain the absence of base-specific fluorochrome bands in the chromosomes of tilapia.

The ultrastructure of the intestinal epithelium in Oreochromis niloticus on a diet with different concentrations of zinc

The effects of diets with variable zinc levels on the midgut epithelial cells were studied in Oreochromis niloticus L. One hundred and twenty fry of tilapia were apportioned into 4 experimental groups (I, II, III and IV groups), with 30 fish in each treatment, 5 replicate aquaria per treatment containing 6 fish each. The animals of the 4 groups were fed with isonitrogenous (30% crude protein) and isoenergetic (3000 Kcal/Kg of digestible energy) diets with increasing quantities of zinc (44.59; 149.17; 309.93; 599.67 mg Zn/kg of diet), twice a day, for 93 days. Three fish from each group were sacrificed at 36, 66 and 93 days and samples of midgut were removed for ultrastructural analysis. After 93 days of treatment, 3 animals of each experimental group were used for the analysis of zinc concentration by atomic absorption spectrophotometry. The comparative relative index (CRI) revealed that the animals in groups II, III and IV contained, respectively, 1.99%, 34.67% and 22.78% more zinc than the mean concentration in animals from group I. The ultrastructural analysis showed enterocytes with swelling of smooth surfaced endoplasmic reticulum and dilated mitochondria with variable matrix rarefaction and cristae number reduction in the fish exposed to 599.67 mg Zn/Kg of diet at 66 and 93 days of treatment.

Morphological aspects of muscle regeneration in the Nile tilapia (Oreochromis niloticus)

The aim of our study was to analyze the morphological events in the skeletal muscle of the Nile tilapia (Oreochromis niloticus) after a traumatic lesion. Thirty-two fish were used, on which a small longitudinal incision was made in the muscle. The fish were sacrificed after 7, 14, 21, and 42 days and muscle samples were collected from the lesion and processed for morphological analysis. Muscle regeneration in the tilapia occurred gradually through the analyzed period, possibly due to the proliferation and differentiation of myosatellite cells, which were more morphologically evident 7 and 14 days after lesion.

Nile tilapia, Oreochromis niloticus, productive performance and carcass characteristics as related to food management

Male juveniles of Nile tilapia, Oreochromis niloticus, were stocked in 12, 300-m 2 ponds at a density of 1.7 fish/m 2 to evaluate the effects of different food management methods (natural foods and diets in pellet, floating or minced form) on fish production and carcass characteristics. Water quality variables monitored during the period were within acceptable levels for the species. Total fish production was significantly different (P < 0.05) and the highest values were obtained with diets in pellet (5,997 kg/ha) and floating (5,441 kg/ha) form. The fish fed diets had higher contents of body fat (1.57 to 1.98%) and visceral fat (12.64 to 25.04%) than fish fed natural food, which had levels between 0.17 and 0%, respectively. Natural food treatment yielded lower fish production and fish with lower body fat, while treatments that yielded higher fish production (rations) higher percentage of visceral fat. © 2004 by The Haworth Press, Inc. All rights reserved.

Energia digestível para larvas de tilápia-do-nilo (oreochromis niloticus) na fase de reversão sexual

This experiment was conducted to evaluate different levels of digestible energy on the performance of Nile tilapia larvae (Oreochromis niloticus) during the sexual reversion phase. Three hundred and seventy-five larvae with initial average length and weight of 21.0±4.0 mg and 1.19±0.72 cm, respectively, were allotted to 25 30L-aquarium. A completely randomized design with five treatments and five replicates was used. The aquarium with 15 larvae was the experimental unit. The diets were formulated to contain levels of 3,300, 3,525, 3,750, 3,975, and 4,200 kcal/kg of digestible energy and to be isoprotein (38.6% digestible protein). The animals were fed ad libitum five times a day. At the end of the experiment, the averages of weight (PF), survival rate (SO), condition factor (FC) and length (CF) were analyzed. Linear reduction of fishes PF and CF, as the dietary digestible energy levels increased, was observed. No differences on fish SO and FC of different treatments were observed. Deleterious effects on performance due to the increasing dietary digestible energy levels were noticed for Nile tilapia larvae during the sexual reversion.

Tocopherol in the lipid stability of tilapia (Oreochromis niloticus) hamburgers

Presence of tocopherol is effective for fish preservation during frozen storage, inhibiting lipid degradation by oxidation. This work evaluated the antioxidant effects of α-tocopherol in diet and postmortem addition on the final quality of hamburgers produced from tilapia fillets kept frozen for zero, 30, 60, and 90 days. Chemical composition varied within the values found for tilapia fish. The increase in α-tocopherol levels reduced the values of thiobarbituric acid reactive substances (TBARS) in the samples at all time intervals. Tocopherol supplementation in diets protected the hamburgers from lipid oxidation more effectively than postmortem addition. © 2007 Elsevier Ltd. All rights reserved.