Dietary effect of multi-strain prebiotics and probiotics on growth, hemato-biochemical parameters, intestinal histomorphometry, and resistance to hypoxia in juvenile tambaqui (Colossoma macropomum).

This study evaluated the effects of a multi-strain prebiotics and probiotics on the diet of tambaqui Colossoma macropomum. One hundred and fifty juvenile tambaqui (20.2 ± 3.6 g and 10.32 ± 5.78 cm) were randomly distributed in 15 experimental units with a volume of 80 L and fed for 45 days with a diet containing the symbiotic additive at five inclusion levels (0, 2, 4, 6 and 8 g kg feed- 1). At the end of the period, growth performance, survival, hemato-biochemical and intestinal parameters, as well as the fish's resistance to stress were evaluated. The additive did not alter the growth performance, hemato-biochemical and intestinal parameters of the tambaqui. However, the hemato-biochemical parameters of aspartate aminotransferase (AST), plasma protein, hematocrit, glucose, triglycerides and cholesterol showed differences between treatments after the stress challenge. AST increased significantly during challenge and post-challenge. Plasma protein increased significantly during and after the challenge. The hematocrit was highest at 48 h after the challenge and lowest in the 2 g, 6 g and 8 g groups. Glucose was significantly reduced 24 h after the challenge, while triglycerides were lower 24 h and 48 h after the challenge. Cholesterol increased significantly in the challenge. There was an interaction between the factors sampling time and symbiotic concentration for hematocrit and glucose. In hypoxic stress situations, the 2 g concentration was more favorable for the fish. We recommend further studies with 2 g kg feed- 1 in trials lasting more than 45 days.

Edwardsiella tarda in Tambaqui (Colossoma macropomum): A Pathogenicity, Antimicrobial Susceptibility, and Genetic Analysis of Brazilian Isolates.

Edwardsiella tarda is a crucial pathogenic bacterium in tropical aquaculture. This bacterium was recently isolated from tambaqui (Colossoma macropomum), a commercially important fish species in Brazil. This study assessed the antimicrobial susceptibility, pathogenicity, and genetic diversity of the tambaqui-derived E. tarda isolates. Fourteen bacterial isolates isolated from tambaqui were identified as E. tarda by using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry and dnaJ gene sequencing. Antimicrobial susceptibility tests were conducted against seven drugs using the disc diffusion assay. The pathogenicity test conducted by intraperitoneal injection of 2.4 × 107 colony-forming units (CFU) fish-1 of E. tarda (ED38-17) into tambaqui juveniles eventually revealed that neither clinical signs nor death were present. However, splenomegaly and whitish areas in the spleen and kidneys were observed. The histological investigation also revealed granulomatous splenitis, nephritis, and hepatitis occurring internally. Repetitive extragenic palindromic-PCR fingerprinting separated the 14 isolates into three genetic groups. The antibiogram revealed that all E. tarda isolates were wild-type (WT) to florfenicol (FLO), norfloxacin (NOR), neomycin (NEO), erythromycin (ERY), and oxytetracycline (OXY); however, some were non-wild-type to sulfamethoxazole/trimethoprim (7.1%) and amoxicillin (21.4%). Therefore, through experimental infection, E. tarda ED38-17 could induce pathogenic effects in C. macropomum. Additionally, three distinct genetic types were found, and the E. tarda isolates were WT to FLO, NOR, NEO, ERY, and OXY. These findings raise awareness of a bacteria causing unseen lesions, a pathogen that will potentially impact tambaqui aquaculture in the future.

Oogenesis and in vitro reproduction of the twospot astyanax Astyanax bimaculatus (Linnaeus, 1758) exposed to conspecific alarm substance.

Stress situations can be essential to trigger reproduction in fish; however, it may also inhibit it. One of those situations involves the release of the conspecific alarm substance (CAS), a natural stressor, into the water by specific fish epidermal cells after a predator attack. Little is known about the effects of that substance on fish reproduction. This study aimed to evaluate the effects of CAS exposure on the oogenesis and reproduction of the twospot astyanax Astyanax bimaculatus before the hormonal induction for artificial reproduction. No macroscopic or cellular changes in the ovaries were observed for the females exposed to CAS, and the oocyte stages show all females in the same phase of maturation (Spawning Capable). Females exposed to CAS spawned 20 min before the females without exposure. On the other hand, they ovulated only once, whereas the females from the control group ovulated multiple times for approximately two hours after hormonal induction. Moreover, the precocious ovulation of the females submitted to CAS did not generate offspring, since all generated zygotes did not develop. In contrast, the control group females produced more than 11 thousand healthy larvae. Exposing the female fish to CAS during their reproductive management in captivity may reduce breeding success.

Feeding behaviour, locomotion rhythms and blood biochemistry of the neotropical red-tail catfish (Phractocephalus hemioliopterus).

The study evaluated the feeding behaviour of Phractocephalus hemioliopterus through the animals' ability to adapt to the self-feeding system, their preferred feeding times and locomotor activity, as well as the blood biochemistry of juveniles fed in a light/dark cycle. The study was carried out through two experiments, the first of which contained two phases. In experiment 1 - phase I, 24 juveniles (35.28 ± 0.62 g) were distributed in eight 48 l tanks. The tanks were equipped with a self-feeding system and the experiment consisted of evaluating whether the animals were able to adapt to the self-feeding system, as well as evaluating the preferred feeding times and locomotor activity of these animals. A feeding challenge to the animals was introduced in phase II, based on the results of phase I. The results of the first phase evidenced a nocturnal feeding preference. Thus, the feeding challenge consisted of measuring whether the animal would feed during the day and how long it would take to adapt. When the animals consumed 100% of the amount of feed provided daily, phase II was ended. In experiment 2, 24 juveniles of P. hemioliopterus (182.00 ± 14.03 g) were distributed in eight 96 l tanks. This experiment consisted of two treatments with four repetitions, one with exclusive feeding during the middle of the light cycle and another with exclusive feeding in the middle of the dark cycle. At the end, blood samples were collected from the animals for blood biochemistry evaluations. In experiment 1 - phase I, the results showed that the fish adapted very well to the self-feeding system and had a strictly nocturnal feeding behaviour and locomotor rhythm. When they were submitted to the feeding challenge in phase II, the feed intake was stabilized from the 17th day onwards, proportionally to the nocturnal consumption observed in the first phase, thus demonstrating feeding plasticity. In experiment 2, the feeding times influenced the animals' biochemical parameters. Animals fed during the night had higher values of cholesterol and triglycerides than animals fed during the day. It is concluded that P. hemioliopterus has fast adaptability to a self-feeding system, with strictly nocturnal feeding and locomotor behaviours. However, it has feeding plasticity, adapting its behaviour according to food availability. Blood biochemical parameters are influenced by the light/dark feeding cycle.

Mercury metalloproteomic profile in muscle tissue of Arapaima gigas from the Brazilian Amazon.

Metalloproteomics is an innovative methodology for identifying of protein-associated mercury. Thus, we analyzed the muscle proteome of Arapaima gigas (pirarucu), collected in the Madeira River of the Brazilian Amazon, to identify protein-associated mercury, with the aim of identifying possible mercury biomarkers in fish muscle tissue. After obtaining the protein pellet, we conducted two-dimensional electrophoresis (2D PAGE) to fractionate the muscle proteome. Total mercury in muscle tissue and protein pellets and mapping of mercury content in protein spots of the 2D PAGE gels was determined using graphite furnace atomic absorption spectrometry (GFAAS). The protein-associated mercury identification was performed using liquid chromatography coupled with sequence mass spectrometry (LC‒MS/MS). Total mercury determinations by GFAAS indicated concentrations on the order of 153 ± 1.90 mg kg-1 and 142 ± 1.50 mg kg-1 (total precipitation of protein fraction) and 139 ± 1.45 mg kg-1 (fractional precipitation of protein fraction) in muscle tissue and protein pellets, respectively. Mercury concentrations in the range of 48 ± 0.90 to 165 ± 3.00 mg kg-1 were found in twelve protein spots. Among the 2D PAGE protein spots, eleven Hg-binding proteins were identified using LC‒MS/MS, which showed characteristics of mercury exposure biomarkers for important metabolic functions, such as five parvalbumin isoforms, triosephosphate isomerase, cofilin 2 (muscle), and fructose-bisphosphate aldolases.

Chromosomal Diversification in Ancistrus Species (Siluriformes: Loricariidae) Inferred From Repetitive Sequence Analysis.

The Ancistrus genus has extensive chromosomal diversity among species, including heteromorphic sex chromosomes occurrence. However, studies have been shown that chromosomal diversity may still be underestimated. Repetitive sequences represent a large part of eukaryotic genomes, associated with mechanisms of karyotypic diversification, including sex chromosomes evolution. This study analyzed the karyotype diversification of two Ancistrus species (Ancistrus sp. 1 and Ancistrus sp. 2) from the Amazon region by classical and molecular chromosomal markers. Conventional chromosome bands and fluorescence in situ hybridization using probes 18S and 5S rDNA, besides (CA)n, (CG)n, (GA)n, (CAC)n, (CAG)n, (CAT)n, (GAA)n, (GAC)n, (TAA)n, and (TTAGGG)n in tandem repeats were determined on the karyotypes. Ancistrus sp. 1 and Ancistrus sp. 2 presented karyotypes with 2n = 38 (20 m + 14sm+4st, XX/XY) and 2n = 34 (20 m + 14sm, without heteromorphic sex chromosomes), respectively. Robertsonian rearrangements can explain the diploid number difference. C-bands occurred in pericentromeric regions in some chromosomes, and a single 18S rDNA locus occurred in both species. The 5S rDNA showed variation in the number of loci between species karyotypes, suggesting the occurrence of unstable sites and rearrangements associated with these sequences in Ancistrus. The microsatellite mapping evidenced distinct patterns of organization between the two analyzed species, occurring mainly in the sex chromosomes in Ancistrus sp. 1, and in the centromeric and pericentromeric regions of chromosomes m/sm in Ancistrus sp. 2. These data shows the extensive chromosomal diversity of repetitive sequences in Ancistrus, which were involved in Robertsonian rearrangements and sex chromosomes differentiation.

Chloramine-T application for Trichodina sp. in Arapaima gigas juveniles: Acute toxicity, histopathology, efficacy, and physiological effects.

Trichodinids are problematic ectoparasites for intensive arapaima fish farming, which can make their breeding costly and unprofitable. The search for effective and safe therapeutic molecules is constant, and chloramine-T has stood out as an effective biocide to treat various pathogens in aquaculture. Here we investigated the acute toxicity (LC50-4h) of chloramine-T and its behavioural, morphological, histological, physiological, and antiparasitic effects on the control of trichodinids in arapaima juveniles. Initially arapaima were exposed to chloramine-T (0, 10, 20, 40, 50, 60, 80, 100 mg L-1) for 4 h. During this period, behavioural, morphological, and mortality changes were recorded. Immediately after a toxicity test, gills were collected for the histological analysis. The LC50-4h was calculated at 23.8 mg L-1 of chloramine-T. Behavioural changes like erratic swimming, loss of balance, gasping at the water surface, excessive mucus, jumping at the water surface, lethargy and gill hyperventilation, and morphological changes like loss of scales, ocular opacity, and skin darkening, were considered mild in arapaima exposed to 10 and 20 mg L-1 of chloramine-T for 4 h. The arapaima gills exposed to 10, 20 and, 40 mg L-1 of chloramine-T presented epithelial hyperplasia, dilated venous sinus and eosinophilic infiltrate, but they were reversible and of a low pathological degree. Based on the acute toxicity test (LC50-4h) results, concentrations were defined for the in vivo efficacy test, during which arapaima were exposed to 0, 10, 15, and 20 mg L-1 of chloramine-T for 1 h. After a therapeutic bath, physiological and parasitological analyses were performed. The use of 20 mg L-1 of chloramine-T significantly reduced (p < 0.05) parasitism by Trichodina sp. with an efficacy of 50.27 % and 53.23 % in gills and on the body surface, respectively. Chloramine-T did not change haematological parameters (erythrogram and thrombogram) or the biochemical parameters of arapaima after 1 h of exposure. However, monocytosis and neutrophilia were noted at slightly higher concentrations (15 and 20 mg L-1), but changes in fish homeostasis were not characterised. Thus, we demonstrate that the application of a therapeutic bath with 20 mg L-1 of chloramine-T for 1 h reduces parasitism by trichodinids and does not affect general arapaima juvenile health.

Changes in gill and air-breathing organ characteristics during the transition from water- to air-breathing in juvenile Arapaima gigas.

The obligate air-breathing Amazonian fish, Arapaima gigas, hatch as water-breathing larvae but with development, they modify their swim bladder to an air-breathing organ (ABO) while reducing their gill filaments to avoid oxygen loss. Here, we show that significant changes already take place between 4 weeks (1.6 g) and 11 weeks (5 g) post hatch, with a reduction in gill lamellar surface area, increase in gill diffusion distance, and proliferation of the parenchyma in the ABO. By using a variety of methods, we quantified the surface area and diffusion distances of the gills and skin, and the swim bladder volume and anatomical complexity from hatch to 11-week-old juveniles. In addition, we identified the presence of two ionocyte types in the gills and show how these change with development. Until 1.6 g, A. gigas possess only the H+ -excreting/Na+ -absorbing type, while 5-g fish and adults have an additional ionocyte which likely absorbs H+ and Cl- and excretes HCO3- . The ionocyte density on the gill filaments increased with age and is likely a compensatory mechanism for maintaining ion transport while reducing gill surface area. In the transition from water- to air-breathing, A. gigas likely employs a trimodal respiration utilizing gills, skin, and ABO and thus avoid a respiratory-ion regulatory compromise at the gills.

Metalloproteomic Strategies for Identifying Proteins as Biomarkers of Mercury Exposure in Serrasalmus rhombeus from the Amazon Region.

This manuscript describes the results of a metalloproteomic study of mercury in samples of muscle and liver tissue of the species Serrasalmus rhombeus, popularly known as black piranha and characterised as the most voracious and aggressive predator in the Brazilian Amazon. The metalloproteomic study involved using two-dimensional electrophoresis (2D PAGE) to fractionate the proteome of the muscle and liver tissue samples, along with atomic absorption spectrometry in a graphite furnace (GFAAS) to identify mercury associated with protein SPOTs and mass spectrometry with electrospray ionisation (ESI-MS/MS) to characterise the mercury-binding proteins. The protein SPOTs characterised showed concentrations in the order of 156 mg kg-1, which ranks as the highest concentrations of mercury determined so far in metalloproteomic studies involving fish species in the Amazon region. Based on FASTA sequences of proteins characterised by ESI-MS/MS, bioinformatics studies were performed that allowed identifying nine proteins with characteristics of biomarkers of mercury exposure. Of those proteins, glutathione peroxidase stands out as an enzyme of great importance in the antioxidant defence of organisms subjected to oxidative stress caused by xenobiotics.

Acute exposure to environmentally relevant concentrations of copper affects branchial and hepatic phosphoryl transfer network of Cichlasoma amazonarum: Impacts on bioenergetics homeostasis.

The toxic effects of copper (Cu) are linked to dysfunction of metabolism and depletion of adenosine triphosphate (ATP). Nevertheless, the effects related to phosphoryl transfer network, a network of enzymes to precise coupling of the ATP-production and ATP-consuming process for maintenance of bioenergetic, remain unknown. Therefore, the aim of this study was to determine whether the phosphoryl transfer network could be one pathway involved in the bioenergetic imbalance of Cichlasoma amazonarum exposed for 96 h to environmentally relevant concentrations of Cu found in Amazonia water around mines. Branchial mitochondrial creatine kinase (CK) activity was significantly lower in fish exposed to 1500 µg/L Cu than in the control group, while branchial cytosolic CK activity was significantly greater. Branchial (exposed to 750 and 1500 µg/L Cu) and hepatic (exposed to 1500 µg/L Cu) pyruvate kinase (PK) activity was significantly lower in fish exposed to Cu than in the control group. Branchial and hepatic ATP levels were significantly lower in fish exposed to 1500 µg/L than in the control group. Branchial reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to 750 and 1500 µg/L Cu compared to control. Hepatic ROS and LPO levels were significantly higher in fish exposed to 1500 µg/L than in the control group. Branchial and hepatic Cu levels were significantly higher in fish exposed to 1500 µg/L compared to other groups. Exposure to 750 and 1500 µg/L Cu impairs bioenergetics homeostasis, which appears to be mediated by ROS overproduction and lipid peroxidation.

Disturbance of oxidant/antioxidant status and impairment on fillet fatty acid profiles in Brycon amazonicus subjected to acute heat stress.

Amazon fish are vulnerable to climate change. Current temperature increases in Amazonian rivers are likely to continue in the coming years. Elevated temperatures impair homeostasis and subject fish to oxidative stress; nevertheless, the effects of elevated temperature on plasma and muscle oxidative stress as well as fillet fatty acid composition in matrinxã (Brycon amazonicus) remain unknown. Therefore, the aims of the present study were to determine whether antioxidant system is able to withstand acute thermal stress to avoid plasma and muscle oxidative damage and to determine the manner in which matrinxã adjust their muscle fatty acid metabolism in a response to acute heat stress. We exposed juvenile matrinxã to four temperatures (28 °C as control and 30, 32, and 34 °C) for 72 h and observed the effects on plasma and muscle oxidant/antioxidant status and on fatty acid composition. Plasma reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in matrinxã exposed to 34 °C compared with the control group, while they were significantly higher in the muscle of matrinxã exposed to 32 °C and 34 °C compared with the control group. Superoxide dismutase (SOD) activity and total antioxidant capacity (TAC) levels were significantly lower in the plasma and muscle of matrinxã exposed to 34 °C compared with the control group. Plasma glutathione peroxidase (GPx) activity was significantly lower in matrinxã exposed to 32 and 34 °C compared with the control group, while it was significantly lower in the muscle of matrinxã exposed to 34 °C. The total content of saturated fatty acid (SFA) was significantly higher in the fillet of matrinxã exposed to 34 °C compared with the control group, while the total content of polyunsaturated fatty acid (PUFA) was significantly lower. Based on this evidence, we conclude that acute heat stress at 32 °C and 34 °C causes plasma and muscular oxidative stress, and the enzymatic and non-enzymatic antioxidant system is unable to prevent oxidative damage. Moreover, increased levels of total SFA and decreased levels of PUFA occur in an attempt to withstand heat stress and maintain membrane fluidity; nevertheless, these responses can represent negative impacts for consumers.

Effect of seasonal variations on fatty acid composition and nutritional profiles of siluriformes fish species from the amazon basin.

Siluriformes is a highly diverse order of species that exhibit a wide array of morphologies, diets and life strategies. This study evaluated the effect of seasonal variations on the proximate composition, fatty acid composition and nutritional profiles of nine fish species of Siluriformes order from the Amazon Basin. The proximate composition varied throughout seasonal periods with larger differences in total lipid content. Hypophthalmus edentaus showed the largest variation (9.26-26.29%) between drought and flood periods, respectively. The fatty acid composition also varied throughout seasonal periods; the polyunsaturated fatty acid (PUFA) contents significantly (p < 0.05) increased for most of the studied species in the flood period and n-3 fatty acids represented 23% of this increase. Docosahexaenoic acid was the most abundant PUFA during the flood period and Pseudoplatystoma tigrinum (40.17 mg g-1) and Pseudoplatystoma fasciatum (39.93 mg g-1) showed the highest contents. Some species such as Ageneiosus inermis exceed the recommended daily intake of EPA + DHA. All the fish species presented favorable indices of nutritional quality for total lipids. The principal component analysis resulted in a two-principal component model that described 85.15% of the total data variance. It also highlighted that the nine fish species were affected by seasonal variations in their habitat, some to a greater extent than others. The results suggested that the consumption of those fish species contributed to human health.

Ichthyofauna from tributaries of Urubu and Amazonas rivers, Amazonas State, Brazil / Ictiofauna de afluentes dos rios Urubu e Amazonas, Estado do Amazonas, Brasil

Abstract: The Amazonas River basin comprises the world's highest fish species diversity. Anthropogenic interferences in aquatic environments represent a pressure over the maintenance of ecological stability and biodiversity. We inventoried the ichthyofauna of 13 disturbed/modified tributaries of Urubu and Amazonas rivers in the region of the middle Amazon River, between June 2018 and March 2019. A total of 164 species were captured, represented by 11 orders, 37 families and 96 genera. Characiformes was the richest order, followed by Cichliformes and Siluriformes. The most representative families in number of species were Cichlidae, Serrasalmidae, and Characidae. Hemigrammus levis was the most abundant species, and Acarichthys heckelii the most common, registered in all sampled sites. In the present study, species with economic interest were collected, as well as many species recently described and one still waiting for formal description, identified provisionally as Moenkhausia aff. colletii. Therefore, the high fish diversity registered, even in disturbed environments in Middle Amazonas River, denotes the makeable ecological importance of this region for fishes resources and supports the necessity of evaluation of other aquatic environments in the region, as well as the potential impacts on composition, maintenance, and survival of ichthyofauna in environments directly affected by human activities.

Resumo: A bacia do rio Amazonas compreende a maior diversidade de espécies de peixes do mundo. Interferências antropogênicas em ambientes aquáticos representam uma pressão sobre a manutenção da estabilidade ecológica e da biodiversidade. Inventariamos a ictiofauna de 13 afluentes perturbados/modificados dos rios Urubu e Amazonas na região do médio do rio Amazonas, entre junho de 2018 e março de 2019. Foram capturadas 164 espécies no total, representadas por 11 ordens, 37 famílias e 96 gêneros. Characiformes foi a ordem mais rica, seguida por Cichliformes e Siluriformes. As famílias mais representativas em número de espécies foram Cichlidae, Serrasalmidae e Characidae. Hemigrammus levis foi a espécie mais abundante e Acarichthys heckelii a mais comum, registrada em todos os locais amostrados. No presente estudo foram coletadas espécies de interesse econômico, bem como muitas espécies recentemente descritas e uma ainda aguardando descrição formal, identificada provisoriamente como Moenkhausia aff. colletii. Portanto, a alta diversidade de peixes registrada, mesmo em ambientes perturbados no médio rio Amazonas, denota a importância ecológica marcante dessa região para os recursos pesqueiros e suporta a necessidade de avaliação de outros ambientes aquáticos da região, bem como os possíveis impactos na composição, manutenção e sobrevivência da ictiofauna em ambientes diretamente afetados pelas atividades humanas.

Organotropism of total mercury (THg) in Cichla pinima, ecological aspects and human consumption in fish from Amazon region, Brazil.

Specimens of the Cichla pinima are of ecological and economic importance in the Amazon region and are a good bioindicator species of Hg bioaccumulation. Adult specimens of Cichla pinima were obtained by fisheries in the Tapajós river region's impacted gold mining area. Tissues such as muscle, liver, skin, and gill were submitted for total mercury (THg) analysis. For hepatic bioavailability, assays were carried out in the whole liver and in the sub-cellular hepatic fraction. The weight-length relationship showed an equation of W = 2E - 06L3.3002 (R2 = 0.856) indicating an allometric growth. The mean THg values found in the muscle tissue of 676 ± 258 µg kg-1 were below the maximum limit established for human consumption, but were similar to Cichla fish from other areas impacted by Hg in the Amazon region. The trends of levels in the tissues were as follows: liver>muscle>gonads>skin>gill, with no significant differences between the gills and skin. A significant and positive correlation as rs = 0.65 was obtained between the THg contents in cytosolic fraction and the total hepatic THg (liver homogenate). However, only 7% of the THg were found and were available to the sub-cellular hepatic fraction. This profile can be an indicative of a hepatic cellular transference in fish exposed to high Hg levels in the Amazon region now that low concentrations of Hg have been found in the sub-cellular fraction. However, data of differential THg partition in the subcellular compartments should be considered, since others' hepatic fractions can act in the Hg linkage into the cell.

In Situ Localization of Ribosomal Sites in Peckoltia and Ancistomus (Loricariidae: Hypostominae) from the Amazon Basin.

Loricariidae is a diverse group of fish from the neotropical region, occupying a wide variety of freshwater environments. Cytogenetic data have brought important insights into Loricariidae diversity because they help validate undescribed species as well as our understanding of inter- and intraspecific diversity. However, conventional cytogenetic approaches are limited in their ability to detect variability in some lineages, as seen in the Peckoltia clade, owing to their apparent conserved karyotype. Thus, the aim of this work was to map 5S and 18S ribosomal (rDNA) sites in five species of Peckoltia and one species of Ancistomus from the Amazon basin, and discusses the mechanisms of organization and diversification of these clusters. The species analyzed were found to have 2n = 52 and share KF = 38 m-sm +14st-a chromosomes, except Peckoltia vittata with KF = 34 m-sm +18st-a. Extensive variations in the number and location of 5S and 18S rDNA sites were observed among species. These data indicate that inversions are not the most important events in karyotype evolution in this group, and should prove useful in identifying the species studied here. In addition to inversions, transpositions are important evolutionary events that are involved at least in rDNA clusters spreading in Peckoltia and probably in other species of Hypostominae.

Total Mercury Determination in Muscle and Liver Tissue Samples from Brazilian Amazon Fish Using Slurry Sampling.

This paper presents a slurry sampling method for total mercury determination by graphite furnace atomic absorption spectrometry (GFAAS) in tissue of fish from the Amazon. The tissue samples were lyophilized and macerated, and then the slurry samples were prepared by putting 20 mg of tissue, added to a solution containing Triton X-100, Suprapur HNO3, and zirconium nitrate directly in sampling vials of a spectrometer. Mercury standard solutions were prepared under the same conditions as the slurry samples. The slurry samples and the mercury standard solutions were sonicated for 20 s. Twenty microliters of slurry samples were injected into the graphite tube, which contained an internal wall lined with tungsten carbide. Under these conditions, it was possible to thermally stabilize the mercury up to an atomization temperature of 1700 °C. The method was validated by mercury determination in reference materials DORM-4 and DOLT-4. The LOD and LOQ were 0.014 and 0.045 mg kg-1, respectively, and recovery percentages in relation to the concentration values were certified in the order of 98%.

Ecomorphological correlates of twenty dominant fish species of Amazonian floodplain lakes / Correlações ecomorfológicas de vinte espécies de peixes dominantes em lagos de várzea da Amazônia

Abstract Fishes inhabiting Amazonian floodplain lakes exhibits a great variety of body shape, which was a key advantage to colonize the several habitats that compose these areas adjacent to the large Amazon rivers. In this paper, we did an ecomorphological analysis of twenty abundant species, sampled in May and August 2011, into two floodplain lakes of the lower stretch of the Solimões River. The analysis detected differences among species, which could be probably associated with swimming ability and habitat use preferences.

Resumo Os peixes que habitam os lagos de várzea da Amazônia apresentam morfologias bastante diversas, possibilitando a exploração bem sucedida dos diferentes habitats que formam estas áreas adjacentes aos grandes rios amazônicos. Neste artigo, relatamos os resultados de uma análise ecomorfológica de vinte espécies abundantes, coletadas em maio e agosto de 2011, em dois lagos das várzeas do trecho inferior do rio Solimões. As analises indicaram diferenças entre as espécies como função do atributos ecomorfológicos, provavelmente associadas com a capacidade natatória e com preferências por habitat.

Mercury fractionation in dourada (Brachyplatystoma rousseauxii) of the Madeira River in Brazil using metalloproteomic strategies.

This paper presents the results of mercury fractionation in muscle samples of dourada (Brachyplatystoma rousseauxii) from the JIRAU Hydroelectric Power Plant in the Madeira River Basin in the Amazon region of Brazil. The proteome of the dourada muscle was separated by two-dimensional polyacrylamide gel electrophoresis (2D PAGE). The mercury present in the protein spots was determined by graphite furnace atomic absorption spectrometry (GFAAS) after acid mineralisation in an ultrasound bath. The protein spots in which the presence of mercury was detected were characterised by electrospray ionisation tandem mass spectrometry (ESI-MS/MS) after tryptic digestion. The GFAAS determinations indicated that 65% of the mercury was linked to the protein fraction with a molar mass (Mm) of less than 90 kDa. The mercury concentrations in the seven spots in which this protein fraction was present were in the range of 11.40-35.10 µg kg(-1). Based on the mercury concentrations, it was possible to estimate that the protein spots contained approximately 1-3 mercury atoms per protein molecule. The ESI-MS/MS analysis allowed characterisation of the seven protein spots as the following proteins: protein NLRC5 (molar mass=18.10, pI=6.30); 39S ribosomal protein L36 mitochondrial (molar mass=15.40, pI=8.23); N-alpha-acetyltransferase 20 (Mm=15.95, pI=8.80); Mth938 domain-containing protein (Mm=15.01, pI=9.60); ubiquitin-40S ribosomal protein S27a (Mm=9.80, pI=7.60); parvalbumin alpha (Mm=12.40, pI=3.80) and parvalbumin beta (Mm=13.10, pI=3.45).

Effects of the acute exposition to glyphosate-based herbicide on oxidative stress parameters and antioxidant responses in a hybrid Amazon fish surubim (Pseudoplatystoma sp).

The aim of this study was to investigate the effects of acute glyphosate (active ingredient) exposure on the oxidative stress biomarkers and antioxidant defenses of a hybrid surubim (Pseudoplatystoma sp). The fish were exposed to different herbicide concentrations for 96 h. The thiobarbituric acid-reactive substances (TBARS), protein carbonyls and antioxidant responses were verified. The 15 mg a.pL(-1) of herbicide resulted in the death of 50% of the fish after 96 h. An increase in liver and muscle TBARS levels was observed when fish were exposed to the herbicide. The protein carbonyl content was also increased in the liver (4.5mg a.pL(-1) concentration) and brain (2.25 mg a.pL(-1) concentration). The antioxidant activities decreased in the liver and brain after exposure to herbicide. Levels of ascorbic acid in the liver (2.25 mg a.pL(-1) and 4.5 mg a.pL(-1) concentrations) and brain (2.25 mg a.pL(-1) concentration) were increased post-treatment. Levels of total thiols were increased in the liver and brain (2.25 mg L(-1) and 7.5mg a.pL(-1), respectively). Glyphosate exposure, at the tested concentrations affects surubim health by promoting changes that can affect their survival in natural environment. Some parameters as TBARS and protein carbonyl could be early biomarkers for Roundup exposure in this fish species.

GFAAS determination of mercury in muscle samples of fish from Amazon, Brazil.

In the present study, a simple, rapid and sensitive method was developed for the determination of mercury concentrations in the muscle tissue of fish from the Brazilian Amazon using graphite furnace atomic absorption spectrometry (GFAAS) following acid mineralization of the samples in an ultrasonic cold water bath. Using copper nitrate as a chemical modifier in solution and sodium tungstate as permanent modifier, we were able to attain thermal stabilization of the mercury up to the atomisation temperature of 1600 °C in the GFAAS assay. The calculated limits of detection (LOD) and quantification (LOQ) were 0.014 and 0.047 mg kg(-1), respectively.