Carbohydrate tolerance in Amazon tambaqui (Colossoma macropomum) revealed by NMR-metabolomics - Are glucose and fructose different sugars for fruit-eating fish?

In the present study, two approaches were followed to evaluate the metabolic responses of tambaqui (Colossoma macropomum), a frugivorous species, to intraperitoneal (IP) administration of glucose (GLU) and fructose (FRU) in fed (FED) and 10-day fasted (FAST) fish. Glucose and fructose tolerance tests were performed to assess the carbohydrate utilization and complementary NMR-metabolomics analyses were done to elucidate the impacts of sugar mobilization on the metabolic profile of plasma, liver and muscle. Blood was sampled from FED groups at 0, 3, 6 and 24 h; and at 0 and 24 h from FAST groups. Significant differences were observed in the hyperglycaemic peak between sugars at 3 h (GLU - 13.7 ± 2.0 mM vs. FRU - 8.7 ± 1.1 mM; saline 6.3 ± 0.6 mM) and on the return to normoglycaemia (GLU - 8.5 ± 2.2 mM vs. FRU - 5.2 ± 0.9 mM; saline 4.9 ± 0.6 mM) 6 h after IP on the FRU fish. The NMR-metabolomics approach allowed to conclude that tambaqui seems to be more responsive to the feeding regime (FED vs. FAST) than to the injected sugar (FRU vs. GLU). From the studied tissues, plasma showed no significant variations between feeding regimes at 24 h after IP, while muscle and liver revealed some variations on the final metabolome profile between FED and FAST groups. The metabolome variations between feeding regimes are indicative of changes on the amino acid utilization. Fish from FAST group seem to utilize amino acids as energy source rather than for protein synthesis and muscle growth. Variations on glucose concentration in muscle can also indicate different utilization of the sugars depending on the feeding regime.

Eugenol induces body immobilization yet evoking an increased neuronal excitability in fish during short-term baths.

Several studies have suggested eugenol as a suitable anaesthetic for fish. However, it has also been regarded as a toxic and aversive substance to several aquatic organisms, including fish. This study sought to assess the eugenol-induced behavioural alterations and its seizurogenic potential to fish. Moreover, a distinctive methodology for an in vivo evaluation of the brain activity was also presented. Prior to the evaluation of eugenol-induced responses, fish were exposed to pentylenetetrazole (PTZ), to characterize any seizure-like patterns. Antagonizing responses to PTZ were assessed in fish receiving diazepam (BDZ) and subsequently exposed to PTZ. Tambaqui fish juveniles, Colossoma macropomum (15.8 ± 2.8 g) were used as models and assayed as follows: (i) fish exposed to PTZ (15 mM) and (ii) fish receiving a dose of BDZ (10 mg Kg-1) and later exposed to PTZ (15 mM) (BDZ-PTZ group). Thereafter, fish were evaluated throughout (iii) eugenol exposure at 65 µL L-1 (ethanolic solution) and recovery. Control fish and a vehicle control group (ethanol at 585 µL L-1) were also established. PTZ baths elicited body immobilization preceded by hyperactivity in a stereotyped seizure-like behaviour with increased EEG wave amplitude and frequency. PTZ effects in the brain were attenuated by a pre-administration of BDZ. Upon eugenol exposure, tambaqui had an intense neuronal excitability, showing a clonus-like seizure behaviour, also corroborated by the EEG patterns, which were consistent with a seizure-like response. Responses of eugenol-exposed fish resembled those of the PZT-exposed animals, with epileptiform discharges. EMG was in line with the EEG modulation, showing increased tracing oscillations and higher mean amplitudes in PTZ-exposed fish whereas in BDZ-PTZ group muscle contraction was less frequent and powerful. Fish exposed to eugenol showed initially some muscle activity followed by a loss of muscle tonus over time. In summary, our results showed that upon eugenol exposure, although a time-dependent body immobilization was attained, fish presented an intense neuronal excitability comparable to that evoked by PTZ. Eugenol failed to promote depression of the CNS and therefore may be not suitable to be used for general anaesthesia of C. macropomum. As eugenol could be implicated in seizurogenesis and be potentially toxic to the fish brain, protocols suggesting the broad use of eugenol for short-term anaesthesia or euthanasia of fish should be carefully revised, as it raises important concerns in terms of ethics and fish welfare.

Acoustic homogeneity in the piranha Serrasalmus maculatus.

Different studies suggest some social calls could be used in fish identification if their specificity is unambiguously assessed. Sounds of different populations of piranhas Serrasalmus maculatus Kner, 1858 were recorded to determine their homogeneity between rivers inside a single basin (Araguari and Grande River, upper Paraná River basin) and between separated basins (Amazon and Paraná basins). All fish from the different populations produced sounds with similar acoustic features. Consequently, the populations were not discernible based on individual sound characteristics. This high homogeneity between sounds from different populations indicates their usefulness for conservation projects using passive acoustic monitoring in piranhas. Moreover, it supports the use of acoustic features as complementary key characteristics in taxonomic studies.

Integrated use of biomarkers to evaluate the reproductive physiology of Astyanax fasciatus and Hoplias malabaricus males (Teleostei: Characiformes) in polluted reservoirs.

The reproductive physiology of fish can be changed by the presence of pollutants in the water, which act as endocrine disrupting compounds (EDC). We evaluated the impacts of water contaminants in polluted reservoirs acting as possible EDC on the reproductive physiology of Astyanax fasciatus and Hoplias malabaricus males. We used biomarkers with different levels of biological organization. Hoplias malabaricus adult males were collected in the summer and winter at five different sites in the Tietê River Basin: the Ponte Nova reservoir (PN), considered a reference site due to the low anthropogenic influence; the Billings reservoir (BIL) at two different branches; and the Guarapiranga reservoir (GUA) at two different branches. Astyanax fasciatus adult males were collected at PN and BIL. BIL and GUA are subjected to great anthropogenic action. We analyzed gonadal histomorphology, testosterone (T), 11-ketotestosterone (11-KT), estradiol (E2) plasma levels, and gene expression of hepatic vitellogenin (vtgA) and pituitary follicle stimulating hormone (fshß). In the PN reservoir (reference), the biomarkers analyzed in both species did not differ between the periods analyzed. This is an evidence that the animals keep the same reproductive activity during both seasons. The changes in the plasma concentration of gonadal steroids in both species in polluted reservoirs suggest the presence of EDC compounds in the water and/or adjusts of the physiological setpoint to allow the reproduction in such adverse conditions. The use of vtgA as biomarker suggests the presence of estrogenic compounds, mainly in BIL, but with a more evident response of H. malabaricus. However, even considering physiological changes, both species present testes during the maturation phase that allow the reproduction in an environment with a high degree of pollution.

Aerobic Metabolism Impairment in Tambaqui (Colossoma macropomum) Juveniles Exposed to Urban Wastewater in Manaus, Amazon.

The main purpose of the present study was to investigate the potential use of metabolic parameters as non-specific biomarkers of pollution. The Igarapé do Quarenta is a small urban river crossing an industrial area in the city of Manaus, Amazon, and receives the city wastewater without treatment. The fish tambaqui (Colossoma macropomum) were exposed to water collected from two different sites of that stretch for 96 h. After exposure, routine metabolic rate (RMR) was measured, and fish were euthanized for measurements of electron transport system (ETS) activity, Copper (Cu) and Cadmium (Cd) bioaccumulation and biliary PAHs. Water in the sampling points presented low oxygen and high pH, conductivity, dissolved ions, Cu, Cd and ammonia. Bile concentrations of PAHs were high suggesting industrial pollution. The tambaqui exposed to water from Igarapé do Quarenta showed increased RMR and decreased ETS/RMR suggesting impairment of metabolic fish performance and the potential use of these parameters as biomarkers.

Use of bioacoustics in species identification: Piranhas from genus Pygocentrus (Teleostei: Serrasalmidae) as a case study.

The genus Pygocentrus contains three valid piranha species (P. cariba, P. nattereri and P. piraya) that are allopatric in tropical and subtropical freshwater environments of South America. This study uses acoustic features to differentiate the three species. Sounds were recorded in P. cariba, two populations of P. nattereri (red- and yellow-bellied) and P. piraya; providing sound description for the first time in P. cariba and P. piraya. Calls of P. cariba were distinct from all the other studied populations. Red- and yellow-bellied P. nattereri calls were different from each other but yellow-bellied P. nattereri calls were similar to those of P. piraya. These observations can be explained by considering that the studied specimens of yellow-bellied P. nattereri have been wrongly identified and are actually a sub-population of P. piraya. Morphological examinations and recent fish field recordings in the Araguari River strongly support our hypothesis. This study shows for the first time that sounds can be used to discover identification errors in the teleost taxa.

Ecological impacts of pesticides on Astyanax jacuhiensis (Characiformes: Characidae) from the Uruguay river, Brazil.

Brazilian freshwater ecosystems are continuously exposed to pesticides and domestic sewage. The Uruguay River was chosen for this study because of its international importance, as it flows through Brazil, Argentina, and Uruguay. It receives contaminants such as pesticides and domestic residues. Thus, the aim of this study to assess the accumulation of pesticides in muscle of the fish Astyanax jacuhiensis, its biochemical responses, and the presence of pesticides in water. In total, seven pesticides were registered in water from both river sites. Eight pesticides were detected in fish muscle. The biochemical responses showed that brain lipid peroxidation (LPO) and protein carbonyl (PC) in A. jacuhiensis were higher in the summer. Muscle showed the highest LPO levels in the spring and the highest PC in the summer. Liver LPO and PC levels were higher in the spring and summer. In the gills, the PC was higher in the spring and the LPO in the spring and winter. In the brain and in the gills, glutathione-S-transferase activity was high in the summer and autumn. Catalase activity was lower during the winter and spring. Non-protein thiol (NPSH) levels were lower in the brain in the winter and spring. Muscle tissue showed lower NPSH in the winter (site 1). Liver NPSH showed increased levels in liver in the spring and winter (site 2). The biochemical results clearly is related to pesticides and/or to the presence of other contaminants in the water such as metals or domestic sewage. The accumulation of pesticides in fish muscle added evidence that pesticides have been used in the area surrounding the Uruguay River. In conclusion, the biomarkers assayed in the present study could be used in future investigations considering other sampling sites along Uruguay River.

Tambaqui females (Colossoma macropomum) spawn after hormonal induction with buserelin acetate.

The aim of this study was to evaluate induced reproduction in tambaqui females using buserelin acetate as compared with the traditional treatment regimen with carp pituitary extract (CPE). Reproductive traits of females with a body weight (BW) of 8.47 ±â€¯1.52 kg were evaluated in ten females treated with buserelin acetate at the dose of 0.5 mL/kg BW, in a single application, and in ten females treated with CPE at the dose of 5.5 mg/kg BW, in two applications (10 % and 90 %, with a 12-h interval between applications). Spawning rate did not differ between the females treated with buserelin acetate (40 %) and CPE (40 %). Weight, fertilization rate and hatching rate did not differ between the two treatment groups. Degree-hours (determined as the average temperature multiplied by time, in hours, for spawning after the treatment with the second dose of CPE and after the single treatment with buserelin acetate) for spawning and number of oocytes per gram of gametes collected were greater (P < 0.05) in the females treated with buserelin acetate than in the females treated with CPE. Production index, absolute fecundity and relative fecundity were greater (P < 0.05) in the females treated with CPE. The hormone buserelin acetate promotes reproduction in tambaqui females with there being a similar spawning rate and oocyte quality, however, lesser production indices and fecundity than when there is the conventional treatment regimen imposed with carp pituitary extract.

Yellow-eyed piranhas produce louder sounds than red-eyed piranhas in an invasive population of Serrasalmus marginatus.

Serrasalmus marginatus is a piranha species native from the lower Paraná River basin and has been invasive in the upper Paraná River basin since the 1980s. In piranhas, sounds of different species have different features. The aim of this study was to investigate if the sounds produced by this species could be used to distinguish two morphotypes: red- and yellow-eyed S. marginatus from the Araguari River (upper Paraná River basin). All the temporal and frequency features of the sounds were equivalent in both groups of eye colour; it corresponds to the species-specific signature described for S. marginatus. Nonetheless, the amplitude features were all statistically different between red- and yellow-eyed piranhas. Yellow-eyed specimens produced louder sounds. In different fish species, colour change in eyes can be due to the absence or the presence of a dominant allele. It can also be involved in social rank or during reproduction. Different hormones and neuropeptides can modulate vocal features. It is hypothesized that a mutation or different hormonal concentrations could explain both sound amplitude and eye colour playing a role in animal communication in S. marginatus.

Metabolic adjustment of Pyrrhulina aff. brevis exposed to different climate change scenarios.

The increases in CO2 concentrations and, consequently, temperature due to climate change are predicted to intensify. Understanding the physiological responses of Pyrrhulina aff. brevis to the climatic scenarios proposed by the IPCC (2014) for the next 100 years is of fundamental importance to determine its susceptibility. Thus, the present study aimed to evaluate the effects of the predicted climatic scenarios for the year 2100 on the metabolic adjustments of P. aff. brevis . Specifically, the rate of oxygen uptake, electron transport system capacity, glycogen and lactate content and the role of Na+K+-ATPases and H+-ATPase were evaluated. P. aff. brevis individuals were exposed for 15 days to the simulated climatic scenarios in climate scenario rooms, where temperature and CO2 in the air were controlled. Two rooms were used to simulate the climatic scenarios predicted by the IPCC (2014): moderate (RCP 6; 2.5 °C and 400 µatm CO2 above current levels) and extreme (RCP 8.5; 4.5 °C and 900 µatm CO2 above current levels), in addition to the "control room" that represents the current scenario. There was an increase in the metabolic rate (MO2) in the animals acclimated to the climate change scenarios (RCP 6 and RCP 8.5) compared to the current scenario. These responses showed a typical effect of temperature on energy demand in relation to the increase in temperature and CO2. Our data showed an increase in O2 consumption (MO2), lactate levels and H+-ATPase activity of the animals acclimated to the moderate and extreme climate change scenarios. Such adjustments presented a clear metabolic imbalance, an alteration that may imply challenges for survival, growth, distribution and reproduction in the face of the expected environmental changes for the year 2100.

Temperature and hypoxia on oxidative stress responses in pacu Piaractus mesopotamicus.

The aim of this study was to verify the effects of the interaction between different temperatures and levels of dissolved oxygen in the oxidative stress parameters of pacu juveniles. A total of 81 pacu juveniles (61.7 ± 9.1 g) were exposed to three temperatures (18, 23, and 28 °C), acclimated for a period of 30 days, and then submitted to three levels of dissolved oxygen: control or normoxia (7 mg L-1); moderate hypoxia (4 mg L-1); and severe hypoxia (2 mg L-1) for 12 h. Glutathione-S-transferase (GST) activity, total antioxidant capacity against peroxyl radicals (ACAP), and protein thiol content (PSH) and LPO (lipid peroxidation) [measured by the TBARS] were measured in gill, liver, muscle and brain. The results indicated that the interaction between different temperatures and dissolved oxygen levels caused alterations in the antioxidant system and induced lipid and protein damage in pacu juveniles. In addition, the effects were organ specific. In conclusion, exposure to moderate and severe hypoxia affect oxidative stress parameters and have been shown to be organ-specific in pacu juveniles. The interaction between 23 °C and hypoxia caused greater disturbances in oxidative stress markers, such as PSH in the gills and liver and LPO in the muscle.

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.

Tambaqui (Colossoma macropomum) acclimated to different tropical waters from the Amazon basin shows specific acute-stress responses.

Stress responses in teleosts include the release of hormones into the bloodstream. Their effects depend on the species and on the environmental conditions. The Amazon basin collects waters of diverse chemical composition, and some fish are able to inhabit several of them. However, the effects of these waters on the stress axis are still unknown. Here we show how acute air-exposure differently affects stress biomarkers in tambaqui (Colossoma macropomum), a tropical model species, when acclimated to two Amazonian waters (Rio Negro -RN- water rich in humic acids and poor in ions, and groundwater -IG- with no humic acids and higher concentration of ions). This study described primary and secondary stress responses after air exposure including plasma cortisol, energy metabolites, pH and ions, skin mucus energy metabolites, as well as gills and kidney Na+/K+-ATPase and Na+/H+-exchanger (NHE) activities. Several differences were described in these stress biomarkers due to the acclimation water. The most remarkable ones include increased mucus glucose only in RN-fish, and mucus lactate only in IG-fish after air exposure. Moreover, an inverse relationship between plasma cortisol and Na+ concentrations as well as a direct relationship between plasma ammonia and branchial NHE activity were observed only in RN-fish. Our results demonstrate how important is to study stress responses in fish acclimated to different environments, as physiological differences can be magnified during episodes of high energy expenditure. In addition to having a direct application in aquaculture, this study will improve the management of critical ecosystems such as the Amazon.

Impact of activation solutions on fresh and frozen-thawed sperm motility and fertilization success for two species of migratory freshwater fishes.

Extracellular environment conditions, ionic concentration, pH, osmolality, and temperature influence sperm activation and sperm quality. The aim of this study was to evaluate the effects of different activating solutions on sperm quality and fertilization rate of fresh and post-thaw sperm in Brycon orbignyanus and Prochilodus vimboides. Activation solutions with ions: NaCl, KCl, CaCl2 (150 mOsm kg-1), without ions: glucose (150 mOsm kg-1) and water: reverse osmosis (RO) (∼0 mOsm kg-1) and tank water (TW) were tested. In experiment 1, fresh sperm motility was activated in each activating solution and motility rate (%), motility quality score (0-5), and motility duration (seconds) were subjectively evaluated using a microscope. In experiment 2, sperm was cryopreserved and post-thaw sperm quality was assessed in each activating solution. Methyl glycol was used as cryoprotectant and for B. orbignyanus a solution of BTS® 5% in water reverse osmosis - 325 mOsm kg-1 was used as extender, while for P. vimboides it was used a solution of glucose 5% in water reverse osmosis - 325 mOsm kg-1. In straw, cryoprotectant, extender, sperm were respectively 10%, 80% 10% (V/V). B. orbignyanus fresh sperm activated in NaCl, KCl, glucose solutions, TW and RO yielded higher averages for all the subjective parameters analysed. In fresh sperm of P. vimboides the highest values were observed when glucose solution was used for sperm motility activation, and the highest fertilization rates were observed in samples activated in glucose or RO solutions. B. orbignyanus post-thaw sperm activated in TW (45.1%) or RO (39.7%) presented the highest values for motility. The highest values of curvilinear velocity (VCL) were observed using glucose (69.5 µm s-1), NaCl (67 µm s-1) and KCl (68.4 µm s-1), but the highest fertilization rates were observed when glucose (3.6%), RO (3.5%) and TW (2.5%) were used. P. vimboides post-thaw sperm activated in glucose solution presented the highest motility rate (41%), VCL (43.7 µm s-1), fertilization rate (18.2%) and hatching rate (13.7%). In order to achieve the best seminal quality, fertilization and hatching rates in both fresh and post-thaw sperm, the glucose solution, TW and RO are indicated for use as sperm motility activators in B. orbignyanus, whereas for P. vimboides the glucose solution and RO are indicated.

Effects of cold stress on juvenile Piaractus mesopotamicus and the mitigation by ß-carotene.

This study investigated the effects of cold stress on morphometrical and hematological biomarkers, energy metabolism, and oxidative stress in different tissues of P. mesopotamicus, and the protective role of ß-carotene. Fish were fed with a control diet (CD) and the same diet supplemented with 105 mg/kg ß-carotene (BD) for 60 days. After the feeding trial, fish fed CD or BD diets were exposed to control (24 °C) and low temperature (14 °C) for 24 h. Fish (CD and BD) exposed to thermal stress showed lower hepatosomatic index. The hemoglobin increased only in CD-fed fish exposed to 14 °C. Increased glycemia, plasmatic protein depletion, and decreased hepatic glycogen were observed in fish fed the CD, while only the lipid levels in liver were augmented in BD-fed fish exposed at 14 °C. Regarding the oxidative stress, increased antioxidant enzymes activity and lipid peroxidation were observed in CD-fed fish exposed to cold. The two-way ANOVA showed an interaction between dietary treatment and temperature for glucose and oxidative stress biomarkers, with the highest values recorded in 14 °C-exposed fish fed with the CD. Our study demonstrated that cold stress had the greatest impact on fish oxidative status, and ß-carotene reduces harmful effects induced by cold in P. mesopotamicus.

Biotransformations, Antioxidant System Responses, and Histopathological Indexes in the Liver of Fish Exposed to Cyanobacterial Extract.

Radiocystis fernandoi, a microcystin (MC) producer, has been common in cyanobacterial blooms in tropical regions. Microcystin is a hepatotoxin that causes tissue damage and even death in animals, including humans; its detoxification process may involve biotransformation and activation of the antioxidant defense system. We evaluated the detoxification pathway, examined the antioxidant defense system responses, and determined the alterations and the organ histopathological indexes in the liver of the tropical fish Hoplias malabaricus after acute and subchronic intraperitoneal exposure to microcystin. The crude microcystin extract of R. fernandoi had predominantly MC-RR and MC-YR. The detoxification process was activated by increasing ethoxyresorufin-O-deethylase activity, whereas glutathione S-transferase was inhibited. The activity of the antioxidant defense enzymes superoxide dismutase (SOD) and glutathione peroxidase decreased after acute exposure; the SOD-catalase system and the glutathione level increased after subchronic exposure. The carbonyl protein level, lipid peroxidation (LPO), and DNA damage were unchanged after acute exposure, whereas protein carbonyl was unchanged, LPO decreased, and DNA damage increased after subchronic exposure. Histopathological alteration indexes differed between acute and subchronic exposure, but the histopathological organ indexes indicate liver dysfunction in both exposure periods. We conclude that MC-RR and MC-YR induce different liver responses depending on the time of exposure, and the antioxidant defense responses after subchronic exposure may help to partially restore the liver function. Environ Toxicol Chem 2020;39:1041-1051. © 2020 SETAC.

Microplastics and copper effects on the neotropical teleost Prochilodus lineatus: Is there any interaction?

Microplastics (MP) are emerging contaminants widely found in aquatic ecosystems. In addition to MP toxicity itself, there is increasing concern about the MP adsorption capacity and the interactive effects with other contaminants, such as copper. The objective of this research was to investigate the effects of polyethylene microplastic and its association with copper (Cu) in genotoxic, biochemical, and physiological biomarkers of the neotropical teleost Prochilodus lineatus. Fish were exposed for 24 and 96 h to MP (20 µg L-1) and Cu (10 µg L-1) and MP + Cu. The results showed that MP and Cu, both isolated and in combination, promoted DNA damage in erythrocytes (96 h) and liver cells (24 and 96 h) indicating that MP and Cu are genotoxic. Fish exposed only to Cu (96 h) showed a decrease in lipid peroxidation in the liver despite of the decrease in glutathione content, indicating the efficiency of other antioxidant defenses. Brain acetylcholinesterase was inhibited in the animals from all the treatments. Although MP did not influence on Cu accumulation in tissues, decreased plasma Na+ and Ca2+ (24 h) occurred after the exposure to MP and Cu, isolated and combined. Exposure to MP and MP + Cu resulted in decreased activity of Ca2+-ATPase (24 h). Taken altogether, these results showed that MP and Cu depicted genotoxic, neurotoxic, and physiological effects on P. lineatus, both alone and combined. An interaction between Cu and MP was observed in plasma Ca2+, where the combination of both contaminants caused a greater effect than the contaminants alone.

Reproductive cycle of the tetra Astyanax bimaculatus (Characiformes: Characidae) collected in Amazonian streams.

A histological characterization of gonadal development in the tetra Astyanax bimaculatus was performed, aimed at determining its reproductive cycle in streams localized inside the Amazonian forest. Collections were carried out monthly from August 2017 to July 2018 at the Zoobotânica Foundation of Marabá, PA. Collected specimens were weighed and measured, and their gonads and liver were removed and weighed to calculate gonadosomatic and hepatosomatic indexes. Gonads were fixed and treated for routine histology for light microscopy. Materials were stained with toluidine blue and haematoxylin and eosin. The Amazonian A. bimaculatus species presented two reproductive periods in the year, one at the end of the winter season and another during the summer. Females showed an asynchronous development of their oocytes and only two reproductive phases of development were observed during the whole period 'developing' and 'spawning capable'. Males presented cystic spermatogenesis, with an anastomosing tubular testis containing spermatogonia spread along the germinal epithelium (unrestricted spermatogonial). These morphological characteristics are considered phylogenetically more primitive. Male specimens were observed to have five different phases during the period: immature, initial maturation, mid maturation, final maturation and regression. The huge fluctuations in Amazonian streams was observed, in which water volumes varied considerably across seasons, culminating even in total drought. In spite of this, A. bimaculatus could be found during all seasons, showing its impressive reproductive adaptation to its conditions.

Structure and Function of the Armored Keel in Piranhas, Pacus, and their Allies.

The serrasalmids: piranhas, pacus, and their relatives, are ubiquitous Neotropical fishes with diverse diets, ecologies, and behaviors. Serrasalmids have a bony, serrated keel which lines the underbellies of these fishes, the structure for which the family is named. We examined the diversity and structure of the keel in piranhas and allies using micro-computed tomography scanning in over 30 species of serrasalmids, a third of the species richness for the family, and for 95 total characiform specimens. The keel is highly diverse across serrasalmids, with serrae shape dictating the overall form of the keel. Serrae shape varies considerably among different species and even within keels themselves. The keel morphology can be divided into distinct anterior and posterior regions, as separated by the pelvic fins. Compared to other characiform fishes, serrasalmid skeletons are frequently damaged. Gouging perforations and signs of healing (serrae fusion) are common on the keel. We propose the keel is a defensive structure based on the high incidence of injury (>50%) in our dataset. This is the highest incidence of damage ever recorded in the skeletons of bony fishes. The loss of the anterior keel region in rheophilic taxa suggests competing performance demands and selective pressures on this structure. Competition and aggression among conspecifics or confamilials is a frequently invoked phenomenon for explaining animal weaponry and armor in terrestrial vertebrates. The keel in serrasalmids and other instances of armor in fishes could be complementary study systems for examining competitive rivalry in vertebrates. Anat Rec, 2018. © 2018 American Association for Anatomy.