Sex matters: Gamete-specific contribution of microRNA following parental exposure to hypoxia in zebrafish.

Oxygen availability varies among aquatic environments, and oxygen concentration has been demonstrated to drive behavioral, metabolic, and genetic adaptations in numerous aquatic species. MicroRNAs (miRNAs) are epigenetic modulators that act at the interface of the environment and the transcriptome and are known to drive plastic responses following environmental stressors. An area of miRNA that has remained underexplored is the sex specific action of miRNAs following hypoxia exposure and its effects as gene expression regulator in fishes. This study aimed to identify differences in mRNA and miRNA expression in the F1 generation of zebrafish (Danio rerio) at 1 hpf after either F0 parental male or female were exposed to 2 weeks of continuous (45 %) hypoxia. In general, F1 embryos at 1 hpf demonstrated differences in mRNA and miRNAs expression related to the stressor and to the specific sex of the F0 that was exposed to hypoxia. Bioinformatic pathway analysis of predicted miRNA:mRNA relationships indicated responses in known hypoxia signaling and mitochondrial bioenergetic pathways. This research demonstrates the importance of examining the specific male and female contributions to phenotypic variation in subsequent generations and provides evidence that there is both maternal and paternal contribution of miRNA through eggs and sperm.

Malathion alters the transcription of target genes of the tumour suppressor tp53 and cancerous processes in Colossoma macropomum: Mechanisms of DNA damage response, oxidative stress and apoptosis.

Different classes of pesticides such as fungicides, herbicides, and insecticides, can induce differential expression of genes that are involved in tumorigenesis events in fish, including the expression of tumor suppressor tp53. The degree and duration of the stressful condition is decisive in defining which tp53-dependent pathway will be activated. Herein we evaluate the target genes expression that participates in the regulation pathway of the tumor suppressor tp53 and in the cancerous processes in tambaqui after exposure to malathion. Our hypothesis is that malathion promotes a gene response that is differentially regulated over time, with positive regulation of tp53 target genes related to the apoptotic pathway and a negative regulation of genes that promote antioxidant responses. The fish were exposed to a sublethal concentration of the insecticide for 6 and 48 h. Liver samples were used to analyze the expression of 11 genes using real-time PCR. Overall, the malathion promoted over time increases in tp53 expression and differential expression of tp53 related genes. The exposure resulted in the activation of damage response related genes, caused a positive expression of atm/atr genes. The pro-apoptotic gene bax was up-regulated and the anti-apoptotic bcl2 was down-regulated. Increased expression of mdm2 and sesn1 in the first hours of exposure and no effect on the antioxidant genes sod2 and gpx1 were also observed. We also witnessed an increase in the expression of the hif-1α gene, with no effect on ras proto-oncogene. The extension of this stressful condition accentuated tp53 transcription, and minimized the levels of mdm2, sens1 and bax; however, it down regulated the levels of bcl2 and the bcl2/bax ratio, which indicates the maintenance of the apoptotic response to the detriment of an antioxidant response.

Vitellogenin Ab structure of the amazonian Arapaima gigas.

The vitellogenin is composed by polypeptides that are precursors of egg yolk proteins that provides embryo and larvae nutrition. The mRNA encoding for vitellogenin Ab (Vtg-Ab; 4,536 bp long and 1,512 amino acids) were obtained by RNA-Seq library sequencing of pirarucu gonads. The Vtg-Ab sequences had high homology with Vtgs of other three teleosts species of the order Osteoglossiformes. The transcript of ovarian Vtg was identified based on structural criteria, and so we classify the Vtg of pirarucu as Vtg-Ab due to the truncated or shortened phosvitin (N-terminal end) and phosvitinless domain (C-terminal end). The Vtg-Ab of pirarucu present two major deletions with 133 amino acids in the Lipovitellin I domain and 89 amino acids in the truncated or shortened Phosvitin domain, both located in the N-terminal end region. The three-dimensional (3-D) structure Vtg-Ab protein shows the presence of a typical 4α-helices bundle protein that runs in anti-parallel. In general, the characterization of Vtg-Ab may be the useful elucidation of the hormonal regulation of vitellogenesis and improve the production of pirarucu for broodstock management in aquaculture and preparation of Vtg antibody production (species-specific) for sex identification.

Temporal exposure to malathion: Biochemical changes in the Amazonian fish tambaqui, Colossoma macropomum.

The main toxicity mechanism of organophosphate insecticides such as malathion is the acetylcholinesterase enzyme inhibition. However, fish responses to organophosphates may vary depending on the activation of different defense mechanisms as well as the length of exposure. As such, the evaluation of acetylcholinesterase activity, in combination with the evaluation of biotransformation and antioxidants enzymes levels, is useful for indicating damage in fish exposed to this insecticide. Moreover, evaluating mitochondrial activity might evidence how the hierarchic responses occur in relation to the length of time that the fish is exposed. Therefore, the aim of our study is to evaluate whether the length of exposure to malathion differentially affects the biochemical responses of tambaqui. Our hypothesis is that the physiological alterations due to exposure are time dependent. Fish were exposed to sublethal concentrations of the insecticide during 6, 12, 24, 36, and 48 h. Contrary to expectations, there was no acetylcholinesterase activity inhibition during the experiment, which indicates an absence of neurotoxicity. Phase II biotransformation mechanism was activated early, especially in the liver. Oxidative damage was evident in the first hours of exposure and was concurrent with the activation of antioxidant enzymes. Mitochondrial bioenergetics were differentially affected by the length of exposure. The data suggest that the tambaqui regulates mitochondrial respiration differently over time, seeking to maintain homeostasis and ATP demand, and ensures the activation of response mechanisms, thus minimizing oxidative damage and avoiding the neurotoxicity of malathion.

Hypoxia tolerance in two amazon cichlids: mitochondrial respiration and cellular metabolism adjustments are result of species environmental preferences and distribution.

The amazon fishes' responses to hypoxia seem to be related to the Amazon basin diversity of aquatic environments, which present drastic daily and seasonal variations in the dissolved oxygen concentration. Among these fishes' adaptation to hypoxia, behavioral, metabolic, physiological, and biochemical responses are well known for some species. In this work, we aimed to identify how two different aquatic environments, normoxic forest streams and hypoxic lakes, dictate the responses to hypoxia for two cichlid species, Mesonauta festivus and Aequidens pallidus. In our results, we found that A. pallidus is less tolerant to hypoxia, which seems to be related to this animal's natural normoxic environment. Even though this species modulated the mitochondrial respiration in order to improve the oxygen use, it also showed a lower decrease in metabolic rate when exposed to hypoxia and no activation of the anaerobic metabolism. Instead, M. festivus showed a higher decrease in metabolic rate and an activation of the anaerobic metabolism. Our data reveal that the natural dissolved oxygen influences the hypoxia tolerance and the species' tolerance is related to its ability to perform metabolic depression. The interest results are the absence of mitochondrial respiration influences in these processes. The results observed with A. pallidus bring to light also the importance of preserving the forests, in which streams hold very specialized species acclimated to normoxia and lower temperature. The importance of hypoxia tolerance is, thus, important to keep fish assemblage and is thought to be a strong driver of fish biodiversity.

Genome assembly and annotation of the tambaqui (Colossoma macropomum): an emblematic fish of the Amazon River Basin.

Colossoma macropomum, known as "tambaqui", is the largest Characiformes fish in the Amazon River Basin and a leading species in Brazilian aquaculture and fisheries. Good quality meat and excellent adaptability to culture systems are some of its remarkable farming features. To support studies into the genetics and genomics of the tambaqui, we have produced the first high-quality genome for the species. We combined Illumina and PacBio sequencing technologies to generate a reference genome, assembled with 39× coverage of long reads and polished to a consensus quality value (QV) of 36 with 130× coverage of short reads. The genome was assembled into 1269 scaffolds (a total of 1,221,847,006 bases), with a scaffold N50 size of 40 Mb, where 93% of all assembled bases were placed in the largest 54 scaffolds corresponding to the diploid karyotype of the tambaqui. Furthermore, the NCBI Annotation Pipeline annotated genes, pseudogenes, and non-coding transcripts using the RefSeq database as evidence, guaranteeing a high-quality annotation. A Genome Data Viewer for the tambaqui was produced, which will benefit groups interested in exploring the unique genomic features of the species. The availability of a highly accurate genome assembly for tambaqui provides the foundation for the discovery of novel ecological and evolutionary insights, and is a helpful resource for aquaculture.

Transcriptomic evidences of local thermal adaptation for the native fish Colossoma macropomum (Cuvier, 1818).

Brazil has five climatically distinct regions, with an annual average temperature difference up to 14 ºC between the northern and southern extremes. Environmental variation of this magnitude can lead to new genetic patterns among farmed fish populations. Genetically differentiated populations of tambaqui (Colossoma macropomum Cuvier, 1818), an important freshwater fish for Brazilian continental aquaculture, may be associated with regional adaptation. In this study, we selected tambaquis raised in two thermally distinct regions, belonging to different latitudes, to test this hypothesis. De novo transcriptome analysis was performed to compare the significant differences of genes expressed in the liver of juvenile tambaqui from a northern population (Balbina) and a southeastern population (Brumado). In total, 2,410 genes were differentially expressed (1,196 in Balbina and 1,214 in Brumado). Many of the genes are involved in a multitude of biological functions such as biosynthetic processes, homeostasis, biorhythm, immunity, cell signaling, ribosome biogenesis, modification of proteins, intracellular transport, structure/cytoskeleton, and catalytic activity. Enrichment analysis based on biological networks showed a different protein interaction profile for each population, whose encoding genes may play potential functions in local thermal adaptation of fish to their respective farming environments.

How will farmed populations of freshwater fish deal with the extreme climate scenario in 2100? Transcriptional responses of Colossoma macropomum from two Brazilian climate regions.

Tambaqui (Colossoma macropomum Cuvier, 1818) is an endemic fish of the Amazon and Orinoco basins, and it is the most economically important native species in Brazil being raised in five climatically distinct regions. In the face of current global warming, environmental variations in farm ponds represent additional challenges that may drive new adaptive regional genetic variations among broodstocks of tambaqui. In an experimental context based on the high-emission scenario of the 5th Intergovernmental Panel on Climate Change (IPCC) report, we used two farmed tambaqui populations to test this hypothesis. RNA-seq transcriptome analysis was performed in the liver of juvenile tambaqui from northern (Balbina Experimental Station, Balbina, AM) and southeastern (Brumado Fish Farming, Mogi Mirim, SP) Brazilian regions kept for 30 days in artificial environmental rooms mimicking the current and extreme climate scenarios. Three Illumina MiSeq runs produced close to 120 million 500 bp paired-end reads; 191,139 contigs were assembled with N50 = 1595. 355 genes were differentially expressed for both populations in response to the extreme scenario. After enrichment analysis, each population presented a core set of genes to cope with climate change. Northern fish induced genes related to the cellular response to stress, activation of MAPK activity, response to unfolded protein, protein metabolism and cellular response to DNA damage stimuli. Genes biologically involved in regulating cell proliferation, protein stabilisation and protein ubiquitination for degradation through the ubiquitin-proteasome system were downregulated. Genes associated with biological processes, including the cellular response to stress, MAPK cascade activation, homeostatic processes and positive regulation of immune responses were upregulated in southeastern fish. The downregulated genes were related to cytoskeleton organisation, energy metabolism, and the regulation of transcription and biological rhythms. Our findings reveal the signatures of promising candidate genes involved in the regional plasticity of each population of tambaqui in dealing with upcoming climate changes.

Temperature and food availability alters the physiology and aerobic capacity of tambaqui (Colossoma macropomum).

Fish of the Amazon experience both daily and seasonal variation in temperature and food availability. In the present work, we investigated the influence of nutrient status changes resulting from feeding Colossoma macropomum five flooded forest fruits on aerobic and swimming performance. To assess the effects of diet, three groups of fish were provided different types of food. One group of was provided with commercial feed, another was given fruit feed, and the last group was not fed. The effects of temperature were tested by keeping half of each group of animals at either 26 °C or 33°C for 30 days. After this period, the animals were transferred to either a respiratory chamber or swimming tunnel. Then, tissues were collected for relevant analyses. Results showed that C. macropomum depresses aerobic metabolism when food is withheld (27% reduction in MO2), and observed effects were intensified at the highest temperature (40% reduction in MO2 at 33 °C). Further, increased temperature negatively influenced hematological (Ht, Hb, VCM), biochemical (glucose, triglycerides, cholesterol), hepatossomatic index and enzymatic (MDH and CS in white muscle) parameters. The animals feeding commercial and fruits enriched diets, have a physiological and biological advantage when compared to the unfed animals at both temperatures. Metabolic suppression is an important strategy for maintaining swimming capacity in C. macropomum. It is suggested that even after a long period of fasting, carbohydrates are important energetic substrates.

Ecophysiology, genotoxicity, histopathology, and gene responses of naphthalene injected Colossoma macropomum (Cuvier, 1818) exposed to hypoxia.

The present study aimed to evaluate the biological responses of Colossoma macropomum to naphthalene injection and subsequent hypoxia exposure, emphasizing the expression of the tumor suppressor gene tp53. Tambaquis were intraperitoneally injected with naphthalene (50 mg/kg) and, after 96 hours, the fish were transferred to respirometry chambers and, submitted to progressive hypoxia for the determination of critical PO2. In a subsequent experiment, the fish received an intraperitoneal injection of naphthalene and were kept for 96 hours under normoxia. Successively, fish were challenged with acute hypoxia (PO2

Oxygen-dependent distinct expression of hif-1α gene in aerobic and anaerobic tissues of the Amazon Oscar, Astronotus crassipinnis.

The aquatic habitats of the Amazon basin present dramatic variation of oxygen level, and, to survive such changes, many aquatic animals developed biochemical and physiological adaptations. The advanced teleost Astronotus crassipinnis (Perciformes) is a fish tolerant to hypoxia and known to endure such naturally variable environments. Hypoxia-Inducible factor-1α (hif-1α) is among the most important and studied genes related to hypoxia-tolerance, maintaining regular cellular function and controlling anaerobic metabolism. In the present work, we studied hif-1α expression and related it to changes in metabolic pathways of Astronotus crassipinnis exposed to 1, 3 and 5 h of hypoxia, followed by 3 h of recovery. The results show that A. crassipinnis depresses aerobic metabolic under hypoxia, with a decrease in glycolysis and oxidative enzyme activities, and increases its anaerobic metabolism with an increase in LDH activity coupled with a decrease in oxygen consumption, which indicates an increase in anaerobic capacity. In addition, the animal differentially regulates hif-1α gene in each tissue studied, with a positive relationship to its metabolic profile, suggesting that hif-1α might be one of the most important induction factors that regulate hypoxia tolerance in this species.

Differential survivorship of congeneric ornamental fishes under forecasted climate changes are related to anaerobic potential

Abstract Two Amazonian closely related tetras - cardinal Paracheirodon axelrodi and green neon P. simulans - were artificially acclimatized to environmental chambers mimicking future climate change scenarios (mild, moderate and extreme), using a microcosm facility. P. simulans survived (100%) to all scenarios after 30 days exposure, while P. axelrodi presented decreasing survival percentages according to environmental severity. These differences may be the reflection of distinct natural acclimatization to microhabitats between the species, which differ in thermal conditions. Survival responses might be related to differences in relative gene expression of lactate dehydrogenase (Ldh), suggesting that P. axelrodi anaerobic potential is lower or non-existent compared to P. simulans, not tolerating long-term thermal challenges. Accordingly, increases in temperature and in CO2 levels caused increases in energy demand and resulted in activation of the anaerobic pathway, as demonstrated by the higher enzyme levels measured in head and tail portions of both species. Sustained anaerobic glycolysis is possible when fish live in challenging environments (low oxygen or high temperature). Our results clearly show that P. simulans has a larger scope for survival to higher energy demands due to its increased anaerobic potential compared to P. axelrodi.

Differential survivorship of congeneric ornamental fishes under forecasted climate changes are related to anaerobic potential.

Two Amazonian closely related tetras - cardinal Paracheirodon axelrodi and green neon P. simulans - were artificially acclimatized to environmental chambers mimicking future climate change scenarios (mild, moderate and extreme), using a microcosm facility. P. simulans survived (100%) to all scenarios after 30 days exposure, while P. axelrodi presented decreasing survival percentages according to environmental severity. These differences may be the reflection of distinct natural acclimatization to microhabitats between the species, which differ in thermal conditions. Survival responses might be related to differences in relative gene expression of lactate dehydrogenase (Ldh), suggesting that P. axelrodi anaerobic potential is lower or non-existent compared to P. simulans, not tolerating long-term thermal challenges. Accordingly, increases in temperature and in CO2 levels caused increases in energy demand and resulted in activation of the anaerobic pathway, as demonstrated by the higher enzyme levels measured in head and tail portions of both species. Sustained anaerobic glycolysis is possible when fish live in challenging environments (low oxygen or high temperature). Our results clearly show that P. simulans has a larger scope for survival to higher energy demands due to its increased anaerobic potential compared to P. axelrodi.

Biomarker responses and PAH ratios in fish inhabiting an estuarine urban waterway.

Many cities worldwide are established adjacent to estuaries and their catchments resulting in estuarine contamination due to intense anthropogenic activities. The aim of this study was to evaluate if fish living in an estuarine urban waterway were affected by contamination, via the measurement of a suite of biomarkers of fish health. Black bream (Acanthopagrus butcheri) were sampled in a small urban embayment and a suite of biomarkers of fish health measured. These were condition factor (CF), liver somatic index (LSI), gonadosomatic index (GSI), hepatic EROD activity, polycyclic aromatic hydrocarbon (PAH) biliary metabolites, serum sorbitol dehydrogenase (s-SDH) and branchial enzymes cytochrome C oxidase (CCO), and lactate dehydrogenase (LDH) activities. The biomarkers of exposure EROD activity, and pyrene- and B(a)P-type biliary metabolites confirmed current or recent exposure of the fish and that fish were metabolizing contaminants. Relative to a reference site, LSI was higher in fish collected in the urban inlet as was the metabolic enzyme LDH activity. CF, GSI, s-SDH, CCO, and naphthalene-type metabolites were at similar levels in the urban inlet relative to the reference site. PAH biliary metabolite ratios of high-molecular-weight to low-molecular-weight suggest that fish from the urban inlet were exposed to pyrogenic PAHs, likely from legacy contamination and road runoff entering the embayment. Similarly, the sediment PAH ratios and the freshness indices suggested legacy contamination of a pyrogenic source, likely originating from the adjacent historic gasworks site and a degree of contamination of petrogenic nature entering the inlet via storm water discharge. Biomarkers of exposure and effect confirmed that black bream collected in the Claisebrook Cove inlet, Western Australia, are currently exposed to contamination and are experiencing metabolic perturbations not observed in fish collected at a nearby reference site.

Large-scale SNP discovery and construction of a high-density genetic map of Colossoma macropomum through genotyping-by-sequencing.

Colossoma macropomum, or tambaqui, is the largest native Characiform species found in the Amazon and Orinoco river basins, yet few resources for genetic studies and the genetic improvement of tambaqui exist. In this study, we identified a large number of single-nucleotide polymorphisms (SNPs) for tambaqui and constructed a high-resolution genetic linkage map from a full-sib family of 124 individuals and their parents using the genotyping by sequencing method. In all, 68,584 SNPs were initially identified using minimum minor allele frequency (MAF) of 5%. Filtering parameters were used to select high-quality markers for linkage analysis. We selected 7,734 SNPs for linkage mapping, resulting in 27 linkage groups with a minimum logarithm of odds (LOD) of 8 and maximum recombination fraction of 0.35. The final genetic map contains 7,192 successfully mapped markers that span a total of 2,811 cM, with an average marker interval of 0.39 cM. Comparative genomic analysis between tambaqui and zebrafish revealed variable levels of genomic conservation across the 27 linkage groups which allowed for functional SNP annotations. The large-scale SNP discovery obtained here, allowed us to build a high-density linkage map in tambaqui, which will be useful to enhance genetic studies that can be applied in breeding programs.

Influence of the natural Rio Negro water on the toxicological effects of a crude oil and its chemical dispersion to the Amazonian fish Colossoma macropomum.

The increment in crude oil exploitation over the last decades has considerably increased the risk of polycyclic aromatic hydrocarbon (PAH) contamination to Amazonian aquatic environments, especially for the black water environments such as the Rio Negro. The present work was designed to evaluate the acute toxicity of the Urucu crude oil (CO), the chemically dispersed Urucu crude oil (CO + D), and the dispersant alone (D) to the Amazonian fish Colossoma macropomum. Acute toxicity tests were performed, using a more realistic approach, where fish were acclimated to both groundwater (GW), used as internal control, and natural Rio Negro water (RNW) and exposed to CO, CO + D and D. Then, biomarkers such as ethoxyresorufin-O-deethylase (EROD), superoxide dismutase (SOD), lipid peroxidation (LPO), serum sorbitol dehydrogenase (s-SDH) in liver, DNA damage in blood cells, and the presence of the benzo[a]pyrene-type, pyrene-type, and naphthalene-type metabolites in fish bile were assessed. Fish exposed to CO and CO + D, at both water types tested, presented increased biomarker responses and higher PAH-type metabolites in the bile. However, fish exposed to these treatments after the acclimation to RNW increased the levels of LPO, s-SDH (hepatotoxicity), DNA damage in blood cells (genotoxicity), and benzo[a]pyrene-type metabolites when compared to fish in GW. Our data suggests that some physicochemical properties of Rio Negro water (i.e., presence of natural organic matter (NOM)) might cause mild chemical stress responses in fish, which can make it more susceptible to oxidative stress following exposure to crude oil, particularly to those chemically dispersed.

Transcriptome profiling of two Iberian freshwater fish exposed to thermal stress.

The congeneric freshwater fish Squalius carolitertii and S. torgalensis inhabit different Iberian regions with distinct climates; Atlantic in the North and Mediterranean in the South, respectively. While northern regions present mild temperatures, fish in southern regions often experience harsh temperatures and droughts. Previous work with two hsp70 genes suggested that S. torgalensis is better adapted to harsher thermal conditions than S. carolitertii as a result of the different environmental conditions. We present a transcriptomic characterisation of these species' thermal stress responses. Through differential gene expression analysis of the recently available transcriptomes of these two endemic fish species, comprising 12 RNA-seq libraries from three tissues (skeletal muscle, liver and fins) of fish exposed to control (18 °C) and test (30 °C) conditions, we intend to lay the foundations for further studies on the effects of temperature given predicted climate changes. Results showed that S. carolitertii had more upregulated genes, many of which are involved in transcription regulation, whereas S. torgalensis had more downregulated genes, particularly those responsible for cell division and growth. However, both species displayed increased gene expression of many hsps genes, suggesting that they are able to deal with protein damage caused by heat, though with a greater response in S. torgalensis. Together, our results suggest that S. torgalensis may have an energy saving strategy during short periods of high temperatures, re-allocating resources from growth to stress response mechanisms. In contrast, S. carolitertii regulates its metabolism by increasing the expression of genes involved in transcription and promoting the stress response, probably to maintain homoeostasis. Additionally, we indicate a set of potential target genes for further studies that may be particularly suited to monitoring the responses of Cyprinidae to changing temperatures, particularly for species living in similar conditions in the Mediterranean Peninsulas.

Genomic Resources Notes accepted 1 April 2015 - 31 May 2015.

This article documents the public availability of transcriptomic resources for (i) the stellate sturgeon Acipenser stellatus, (ii) the flowering plant Campanula gentilis and (iii) two endemic Iberian fish, Squalius carolitertii and Squalius torgalensis.

Rapid regulation of blood parameters under acute hypoxia in the Amazonian fish Prochilodus nigricans.

Prochilodus nigricans, locally known as curimatã, is an Amazonian commercial fish that endures adverse environmental conditions, in particular low dissolved oxygen, during its migration. Poorer environmental conditions are expected in the near future. Prochilodus nigricans overcomes current seasonal and diurnal changes in dissolved oxygen by adjusting erythrocytic levels of ATP and GTP, modulators of Hb-O2 affinity. Will this fish species be endangered under more extreme environmental conditions as hypoxia and acidification tend to occur in a shorter period of time? As P. nigricans does not exhibit any apparent morphological alterations to exploit the air-water interface, it must rely on fast adjustments of blood properties. To investigate this aspect, basic hematology indices, pHe, pHi, plasma lactate, erythrocytic levels of ATP and GTP and functional properties of the hemolysate of P. nigricans were analyzed over a period of 6h in hypoxia and subsequent recovery in normoxia. The levels of erythrocytic GTP were four times higher than ATP and were reduced to » of the original level after 3h under hypoxia. Erythrocytic levels of ATP were unaffected over the experimental period. All other analyzed blood parameters exhibited a time-course change in animals under hypoxia and returned to normoxic levels. Considering the hemolysate functional properties and the ability to regulate the above mentioned blood characteristics, P. nigricans is able to endure short-term changes in dissolved oxygen.

Genetic variability of wild and captivity populations of Colossoma macropomum (Cuvier, 1818) / Variabilidade genética de populações selvagens e de cativeiro de Colossoma macropomum (Cuvier, 1818)

Tambaqui (Colossoma macropomum) is among the most important fish species of the Amazon and one of the most cultivated in Brazil. In the present work we have evaluated the genetic variability of wild and captivity populations of C. macropomum. Enzymatic markers were used to estimate the genetic variability of 41 specimens from a wild group; and 30, 33 and 45 from three captivity groups, which came from Pentecostes (Ceará State), Jaboticabal (São Paulo State) and Itacoatiara (Amazonas State), respectively. Nine isoenzymic systems were used to evaluate the genetic variability of these populations. Using zimogram data we obtained the polymorphism level, allele number, allelic frequency, observed and expected heterozigosity, Wright F statistics (FIS, FST), genetic distance, level of similarity and group analysis. The isoenzymic data showed that, from the nine systems, six presented polymorphic loci (Fbp-2, G6pdh-2, G6pdh-3, Pgi-1, Pgi-2 and Pgm-1). The populations from Pentecostes and Jaboticabal presented loss of genetic variability and low heterozigosity, compared to the wild population and to the artificial population acquired at Itacoatiara fish farm. Based on these results and on fish farmer information we could consider the population from Itacoatiara as recently derived from a wild population. Concluding, we suggest that the artificial populations of tambaqui, which contain animals originated from this funding population at Pentecostes, should be renewed with the introduction of a new group of individuals with genetic variability equivalent to the wild population.

O tambaqui é uma espécie de peixe bastante importante na região amazônica e uma das espécies mais cultivadas no Brasil. O objetivo deste trabalho foi avaliar a variabilidade genética do Colossoma macropomum de cativeiro e selvagem, utilizando marcadores isoenzimáticos. Utilizamos 41 indivíduos de uma população da natureza e 30, 33 e 45 de populações de cativeiro de Pentecoste, Jaboticabal e Itacoatiara, respectivamente. Nove sistemas isoenzimáticos foram utilizados para verificar a variabilidade genética do tambaqui, bem como os níveis de polimorfismos, números de alelos, freqüências alélicas, heterozigosidade observada e esperada, estatística F de Wright (FIS e FST), distância e similaridade genética, e análise de agrupamento. Das nove isoenzimas analisadas apenas seis sistemas apresentaram polimorfismo (Fbp-2, G6pdh-2, G6pdh-3, Pgi-1, Pgi-2 and Pgm-1). Verificamos que as populações de Pentecostes e Jaboticabal estão com falta de heterozigotos e apresentam-se estruturadas geneticamente em relação às populações de Itacoatiara e da Natureza que apresentam excesso de heterozigotos e não são estruturadas. Concluímos que os indivíduos de Itacoatiara são provenientes de populações selvagens e sugerimos que se realize uma renovação dos plantéis de tambaqui de Pentecostes e Jaboticabal, com o intuito de recuperar a variabilidade genética perdida.