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.

The correlation between subordinate fish eye colour and received attacks: a negative social feedback mechanism for the reduction of aggression during the formation of dominance hierarchies

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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)

Effects of a small fish cage farm on zooplankton assemblages (Cladocera and Copepoda: Crustacea) in a sub-tropicalreservoir (SE Brazil).

In Brazil, the environmental impacts of fish cage farming in water reservoirs have not been well studied. As this activity is being increasingly practiced, investigations on the impacts of this practice are strongly needed. The goal of this study was to investigate the effects of a small cage tilapia farm on zooplankton assemblages in an oligo/mesotrophic reservoir (Jurumirim Reservoir, Paranapanema River). Zooplankton, limnological variables, and water samples were obtained trimonthly during a year at two sample sites, one was located adjacent to the cage farm and the control area was located one kilometer away from it. Eighteen species were identified and Cladocera was the dominant group. The same species of microcrustaceans were identified at both sites. Among the ecological attributes studied, only evenness showed a tendency towards being higher in the control site. Significant differences between studied variables in the sites were observed only for material in suspension. The results of the study indicate that, during the studied period, the cage farm did not generate detectable changes in the zooplankton assemblages and their ecological attributes. However, small differences in some limnological variables could be an indication of some environmental changes associated with the fish farm system.

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.

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.

Anesthesia of fish with benzocaine does not interfere with comet assay results

Fish blood erythrocytes are frequently used as sentinels in biomonitoring studies. Usually, fish blood is collected by painful cardiac or caudal vein punctures. Previous anesthesia could decrease animal suffering but it is not known at present whether anesthesia can cause confounding effects. Therefore, using the alkaline single cell gel (SCG)/comet assay with blood erythrocytes of the cichlid fish Nile tilapia, we tested for a possible modulation of induced DNA damage (methyl methanesulfonate; MMS) by the anesthetic benzocaine administered by bath exposure (80mg/l for ∼10min). Furthermore, benzocaine (80-600mg/l) was tested for its genotoxic potential on fish erythrocytes in vitro and for potential interactions with two known genotoxins (MMS and hydrogen peroxide). Our results did neither indicate a significant increase in the amount of DNA damage (even after a 48h follow-up), nor indicated interactions with MMS-induced DNA damage when fish were exposed to benzocaine in vivo. There was also no increase in DNA damage after in vitro exposure of fish erythrocytes to benzocaine. Clear concentration-related effects were observed for the two genotoxins in vitro, which were not significantly altered by the presence of benzocaine. These results suggest that anesthesia of fish does not confound comet assay results and the use of blood samples from anesthetized fish can be recommended with regard to animal welfare. © 2002 Elsevier Science B.V. All rights reserved.

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.

Utilization of fish protein hydrolysate in prepared diets for pacu, Piaractus mesopotamicus, larvae

Growth and survival rates of pacu, Piaractus mesopotamicus, larvae fed prepared diets containing different animal protein sources were evaluated. Four diets with the same level of crude protein (CP) (36%) and calories (4.02 kcal gross energy/g of diet) were fed to the larvae. Diets were formulated to contain one of four protein sources: (1) fish meal (FM), (2) tilapia residue silage (TS), (3) protein hydrolysate from tilapia residue (HT), and (4) eviscerated tilapia residue (HET). Larvae were fed Artemia nauplii for six days, prior to the start of the study, and the prepared diet was supplied from day 7 until the study concluded. Variance analysis showed no significant differences (P > 0.05) for survival rates and larval final lengths among treatments. However, final average weights were significantly different (P < 0.05 for larvae fed FM and HT. Average survival rates were relatively high and ranged from 68.1% to 73.9%. After the live food was replaced by prepared diets, no larval growth was observed for any treatment. Fish protein hydrolysate (HT and HET) and fish silage showed potential to be used as ingredients in the diet of pacu larvae. However, hydrolysate inclusion levels, processing methods to minimize nutrient lixiviation, and the best moment to replace live food with an inert diet (weaning) need further investigation. © 2003 by The Haworth Press, Inc. All rights reserved.

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.

Relative bioavailability of zinc in supplemental inorganic and organic sources for Nile tilapia Oreochromis niloticus fingerlings

A 90-day feeding experiment was conducted with sex reversed Nile tilapia (Oreochromis niloticus) fingerlings fed purified or practical diets supplemented with different zinc sources to evaluate fish growth performance and zinc and iron retention in fish bones, fillets, liver, skin and eyes. The relative bioavailability value (RBV) of zinc in the supplemental sources tested was also calculated. Fish were fed with isonitrogenous and isoenergetic purified or practical diets supplemented with 150 mg Zn kg -1, as zinc sulphate monohydrate (ZnSO 4), zinc oxide (ZnO) or zinc amino acid complex (Zn-AA). The feeding trial was conducted in 30, 50 L aquaria where four 0.66 ± 0.01 g (mean ± SD) fingerlings were initially stocked. No significant differences were observed for any growth performance variables (P > 0.05). In practical diets, only ZnSO 4 and ZnO presented bone zinc retention similar to that for the standard zinc source. Zinc concentration in the bone of fish fed practical diet supplemented with Zn-AA (171 ± 3.62 μg g -1) was significantly lower than that verified for the practical diets supplemented with the standard zinc source (200 ± 17.7 μg g -1) or with ZnSO 4 (204 ± 19.9 μg g -1). Assuming the concentration of zinc in bones as the response criterion, the supplemental zinc RBV from ZnSO 4 (105%) was higher than the RBV for Zn-AA (95.1%) or ZnO (94.9%). Iron concentration in the bones of animals fed the non-zinc-supplemented purified diet was significantly higher than that observed for purified diet supplemented with Zn-AA (P < 0,05). The results of the present work allowed us to conclude that ZnSO 4 in relation to ZnO or Zn-AA was the supplemental zinc source with higher zinc bioavailability to Nile tilapia. © 2005 Blackwell Publishing Ltd.

Ventilatory frequency of Nile tilapia subjected to different stressors

Ventilatory frequency (VF) was investigated in the fish Nile tilapia, Oreochromis niloticus, subjected to confinement, electroshock or social stressor. Fish were allowed to acclimatize to tank conditions for 10 days (1 fish/aquarium). VF baseline was determined 5 days after adjustment had been started. At the 10th day of isolation, stressor effects on VF were assessed. The stressors were imposed during 60 min: pairing with a larger resident animal (social stressor), or gentle electroshock (AC, 20 V, 15 mA, 100 Hz for 1 min every 4 min), or space restriction (confinement). VF was quantified immediately after the end of the stressor imposition. Baseline of VF was not statistically different among groups. Social stressor clearly induced VF to increase, while an increased or decreased VF can be observed for both confinement and electroshock. However, fish tend to increase their VF in response to confinement and decrease in the case of electroshock. These results suggest that VF is a sensitive behavioural indicator for distinguishing stress responses in the fish Nile tilapia among different stressors. © 2006 Elsevier GmbH. All rights reserved.