Growth and carbon turnover of Piaractus mesopotamicus Holmberg, 1887 (Osteicthyes: Characidae): contribution of extruded feed and natural food.

Piaractus mesopotamicus, is a fish usually farmed in semi-intensive systems with access to natural food and supplementary feed. This study evaluates effects of feed allowance on the productive performance, carbon turnover and proportions of nutrient (carbon) contribution of feed and natural food for the growth of pacu. Juvenile fish were stocked in fiberglass tanks and fed to 100, 75, 50, 25, 0% apparent satiety (ApS), with a practical, extruded (C4 photosynthetic pathway) feed in a randomized design trial (n=3); plankton production for simulated semi-intensive farming system condition was induced by chemical fertilization. A control treatment was set up in tanks devoid of natural food. Data on muscle stable carbon isotope ratios were used to study carbon turnover using a relative growth-based model. Low variation of the δ13C impaired fitting a turnover model curve for the 0 and 25 % ApS treatments. Fish of the 100% and 75% ApS treatments reached circa 95% and 82.85% of the carbon turnover, respectively. Extruded feed was the main nutrient source for the growth of pacu in the semi-intensive, simulated farming condition. The current study contributes to the knowledge of the relationship between feeding rates and carbon turnover rates in the pacu muscle.

3D spheroid culturing of Astyanax mexicanus liver-derived cell lines recapitulates distinct transcriptomic and metabolic states of in vivo tissue environment.

In vitro assays are crucial tools for gaining detailed insights into various biological processes, including metabolism. Cave morphs of the river-dwelling fish species, Astyanax mexicanus, have adapted their metabolism allowing them to thrive in the biodiversity-deprived and nutrient-limited environment of caves. Liver-derived cells from the cave and river morphs of A. mexicanus have proven to be excellent in vitro resources to better understand the unique metabolism of these fish. However, the current 2D cultures have not fully captured the complex metabolic profile of the Astyanax liver. It is known that 3D culturing can modulate the transcriptomic state of cells when compared to its 2D monolayer culture. Therefore, to broaden the possibilities of the in vitro system by modeling a wider gamut of metabolic pathways, we cultured the liver-derived Astyanax cells of both surface and cavefish into 3D spheroids. We successfully established 3D cultures at various cell seeding densities for several weeks and characterized the resultant transcriptomic and metabolic variations. We found that the 3D cultured Astyanax cells exhibit an altered transcriptomic profile and consequently represent a wider range of metabolic pathways, including cell cycle changes and antioxidant activities, associated with liver functioning as compared to its monolayer culture. Enzymatic assay measuring antioxidants in 2D culture and 3D spheroids also revealed enhanced antioxidative capacity of 3D cultured spheroids, in line with the differential gene expression data. Additionally, the spheroids also exhibited surface and cave-specific metabolic signatures, making it a suitable system for evolutionary studies associated with cave adaptation. Notably, cavefish derived spheroids enriched for genes responding to xenobiotic stimulus, while the ones from surface enriched for immune response, both of which resonated with known physiologically adaptations associated with each morph. Taken together, the liver-derived spheroids prove to be a promising in vitro model for widening our understanding of metabolism in A. mexicanus and of vertebrates in general.

Multiple biomarkers response of Astyanax lacustris (Teleostei: Characidae) exposed to manganese and temperature increase.

The present study aimed to evaluate the toxicity of Mn (6.65 mg/L) at different exposure times (96 h, 7, 14, and 21 days) and evaluate its possible toxic effects on the fish Astyanax lacustris through multi-biomarkers and the maximum critical temperature (CT Max). The results show an increase in the Mn accumulation (liver and gills) with increasing exposure time. The glutathione S-transferase (GST) activity showed differences in the group exposed to Mn for 96 h compared to the group exposed for 21 days. The acetylcholinesterase (AChE) activity increased in the fish exposed for 7 days compared to the control group. On the other hand, no genotoxic changes were observed. The CT Max showed that the loss of equilibrium of 50% of the fish occurs at a temperature of 39ºC, with and without the Mn presence. Furthermore, the catalase gene expression (oxidative stress) did not show alterations.

Oxidative Stress in a Wild Population of the Freshwater Fish Hyphessobrycon Luetkenii Chronically Exposed to a Copper Mining Area: New Insights into Copper Toxicology.

In this study, we examine markers of oxidative stress in the tetra Hyphessobrycon luetkenii collected from two locations in the copper contaminated João Dias creek (southern Brazil). Also, specimens were translocated from a clean reference section of the creek to a polluted stretch and vice-versa. Fish were held at in submerged cages for 96 h and then sacrificed. Nuclear abnormalities in erythrocytes and total antioxidant capacity, lipid peroxidation and protein carbonylation in gills, brain, liver and muscle displayed similar trends in both groups. Lipid peroxidation increased in all tissues of individuals translocated to the polluted site but only in liver and muscle of those translocated to the reference site. Increased protein carbonylation was also observed in gills of individuals translocated to the reference location. These results suggest similar oxidative stress among fish from the reference and polluted locations and that long-term metals exposure may require adaptations toward oxidative stress responses.

Biotransformation and oxidative stress responses in fish (Astyanax aeneus) inhabiting a Caribbean estuary contaminated with pesticide residues from agricultural runoff.

The estuarine ecosystem of Madre de Dios Lagoon (MDL), in the Caribbean Coast of Costa Rica, is exposed to contamination with pesticide residues coming from the upstream agricultural areas. Biomarkers can provide a better indication of the fitness of biota in real mixture exposure scenarios than traditional lethal dose toxicity measurements. Here, we measured biomarkers of biotransformation, oxidative stress, and neurotoxicity on Astyanax aeneus, an abundant fish species in MDL. Glutathione S-transferase activity (GST), catalase activity (CAT), lipid peroxidation (LPO), and cholinesterase activity (ChE) were measured in fish collected during seven sampling campaigns, carried out between 2016 and 2018. Pesticide residues were analyzed in surface water samples collected every time fish were sampled. Residues of 25 pesticides, including fungicides, insecticides, and herbicides, were detected. The biomarkers measured in A. aeneus varied along the sampling moments, with biotransformation and oxidative stress signals showing a coupled response throughout the assessment. Furthermore, significant correlations were established between three biomarkers (GST, LPO, and CAT) and individual pesticides, as well as between GST and LPO with groups of pesticides with shared biocide action. Among pesticides, insecticide residues had a major influence on the responses observed in fish. This work demonstrates the chronic exposure to pesticide residues in MDL and how such exposure is related to physiological responses in fish that can affect their health and potentially, the trophic networks. This early warning information should be considered to improve the protection of estuarine ecosystems in the tropics.

What happens to Hoplias malabaricus fed on live prey (Astyanax altiparanae) previously exposed to copper? A multiple biomarker approach.

Copper waterborne toxicity is well understood in aquatic organisms. However, the dietary copper effects are much less known, especially in tropical fish. The toxicity of copper via the trophic route could be influenced by the composition of the food, and diets naturally impregnated with copper seem to have greater toxicity at lower concentrations than artificially impregnated ones. Thus, our objective was to investigate the effects of copper on juveniles of the Neotropical fish Hoplias malabaricus fed on live prey (Astyanax altiparanae) previously exposed to the metal (20 µg L - 1) for 96 h. The prey fish were given to H. malabaricus every 96 h, totaling 10 doses at the end of the experiment. Thus, after 40 days fish were killed and tissues were sampled. Blood showed to be the only tissue in which copper accumulated. Anemia was found and there was damage to the DNA of erythrocytes. Furthermore, ionic imbalances were observed in plasma. There was an increase in the concentration of Na+ and Cl- and a decrease in Ca2+, which were associated with increased copper uptake in the gastrointestinal tract of fish fed on copper exposed prey. All the antioxidant enzymes evaluated in the gills showed decreased activity compared to the control group. Copper seems to have interfered in the energy metabolism of H. malabaricus, since a lower condition factor and feed conversion efficiency rate were observed in fish fed with copper diet. The present study confirms the trophic route as an important copper toxicity pathway for H. malabaricus and reinforces the idea that metal toxicity can be increased when it is naturally impregnated in the prey tissues, even if the prey has been exposed to the metal only for a short period of time.

Fish biomarker responses reflect landscape anthropic disturbance in savanna streams.

Disturbance in the landscape surrounding streams can interfere with water quality and cause harm to aquatic organisms. In this study, we evaluate the influence of land use on the genetic and biochemical biomarkers of fish in streams of Brazilian savanna (Cerrado). We also evaluated whether biomarker responses are seasonally consistent. For this purpose, individuals of the Neotropical tetra fish Astyanax lacustris were exposed in cages for 96 h, in 13 streams draining agroecosystems with different degrees of disturbance during the dry and wet seasons. After exposure, blood, liver, and gills were collected for multibiomarker analyses (micronuclei, erythrocytic nuclear abnormalities, lipid peroxidation, antioxidant enzymes, and biotransformation enzyme). The results showed that the gradient of anthropic disturbance was positively associated with genotoxic damage (erythrocytic nuclear abnormalities) and negatively associated with antioxidant and biotransformation enzymes of the liver in both seasons. No association of the gradient of anthropic disturbance with the frequency of micronuclei and for most gill enzymes was found for both seasons. Landscape disturbance was also negatively associated with water quality in the wet season. These results indicate that changes in land use interfere with the genetic and biochemical processes of organisms. Thus, the multibiomarker approach may represent an effective strategy for assessing and monitoring terrestrial landscape disturbance.

Does color play a predominant role in the intake of microplastics fragments by freshwater fish: an experimental approach with Psalidodon eigenmanniorum.

Microplastics (MPs) have been reported in fish species from several freshwater environments. However, the mechanisms underlying MPs ingestion by fish are still unclear, although they are important to determine the pathway of MPs along freshwater environments food webs. Here, we investigates a fundamental question of why wild freshwater fish ingest plastic. To address this, we conducted a laboratory experiment to assess MP fragments intake according to color (red, green, yellow, white, black, and blue) by a small omnivorous fish species Psalidodon eigenmanniorum (Characidae). Results showed that yellow and blue were the most consumed fragments, whereas fish avoided white fragments. Although it is not yet clear how plastic coloration relates to the selectivity and feeding of freshwater fish, the visual skills at a species-specific level could plausibly explain why certain colors are attractive or deterrent to a particular fish species. This data set can be used as a screening tool that could help to understand the mechanisms underlying the patterns of plastic ingestion by fish, with special emphasis on the color of plastic particles. Future research on mechanisms MPs intake by fish, also providing a multi-species approach is highly recommended.

Genome-wide analysis of cis-regulatory changes underlying metabolic adaptation of cavefish.

Cis-regulatory changes are key drivers of adaptative evolution. However, their contribution to the metabolic adaptation of organisms is not well understood. Here, we used a unique vertebrate model, Astyanax mexicanus-different morphotypes of which survive in nutrient-rich surface and nutrient-deprived cave waters-to uncover gene regulatory networks underlying metabolic adaptation. We performed genome-wide epigenetic profiling in the liver tissues of Astyanax and found that many of the identified cis-regulatory elements (CREs) have genetically diverged and have differential chromatin features between surface and cave morphotypes, while retaining remarkably similar regulatory signatures between independently derived cave populations. One such CRE in the hpdb gene harbors a genomic deletion in cavefish that abolishes IRF2 repressor binding and derepresses enhancer activity in reporter assays. Selection of this mutation in multiple independent cave populations supports its importance in cave adaptation, and provides novel molecular insights into the evolutionary trade-off between loss of pigmentation and adaptation to food-deprived caves.

Enhanced lipogenesis through Pparγ helps cavefish adapt to food scarcity.

Nutrient availability varies seasonally and spatially in the wild. While many animals, such as hibernating animals or migrating birds, evolved strategies to overcome periods of nutrient scarcity,1,2 the cellular mechanisms of these strategies are poorly understood. Cave environments represent an example of nutrient-deprived environments, since the lack of sunlight and therefore primary energy production drastically diminishes the nutrient availability.3 Here, we used Astyanax mexicanus, which includes river-dwelling surface fish and cave-adapted cavefish populations, to study the genetic adaptation to nutrient limitations.4-9 We show that cavefish populations store large amounts of fat in different body regions when fed ad libitum in the lab. We found higher expression of lipogenesis genes in cavefish livers when fed the same amount of food as surface fish, suggesting an improved ability of cavefish to use lipogenesis to convert available energy into triglycerides for storage into adipose tissue.10-12 Moreover, the lipid metabolism regulator, peroxisome proliferator-activated receptor γ (Pparγ), is upregulated at both transcript and protein levels in cavefish livers. Chromatin immunoprecipitation sequencing (ChIP-seq) showed that Pparγ binds cavefish promoter regions of genes to a higher extent than surface fish and inhibiting Pparγ in vivo decreases fat accumulation in A. mexicanus. Finally, we identified nonsense mutations in per2, a known repressor of Pparγ, providing a possible regulatory mechanism of Pparγ in cavefish. Taken together, our study reveals that upregulated Pparγ promotes higher levels of lipogenesis in the liver and contributes to higher body fat accumulation in cavefish populations, an important adaptation to nutrient-limited environments.

Effects of heat shock on energy metabolism and antioxidant defence in a tropical fish species Psalidodon bifasciatus.

Predictions about global warming have raised interest in assessing whether ectothermic organisms will be able to adapt to these changes. Understanding the physiological mechanisms and metabolic adjustment capacity of fish subjected to heat stress can provide subsidies that may contribute to decision-making in relation to ecosystems and organisms subjected to global climate change. This study investigated the antioxidant defence system and energy metabolism of carbohydrate and protein responses in the gill, liver and kidney tissues of Psalidodon bifasciatus (Garavello & Sampaio 2010), a Brazilian freshwater fish used in aquaculture and in biological studies, following exposure to heat shock at 31°C for 2, 6, 12, 24 and 48 h. The fish presented signs of stress in all tissues tested, as evidenced by increased lipid peroxidation concentration at 2 h and phosphofructokinase, hexokinase and malate dehydrogenase activity at 48 h in the gills; increased glutathione-S-transferase activity at 12 h, citrate synthase activity at 24 h and concentration of reduced glutathione (GSH) concentration at 12 and 48 h in the liver; and through increased activity of superoxide dismutase at 48 h, glutathione reductase at 24 h, glucose-6-phosphate dehydrogenase at 48 h and concentration of GSH at 24 h in the kidney. In the kidneys, changes in the antioxidant system were more prominent, whereas in the gills, there were greater changes in the carbohydrate metabolism. These results indicated the importance of glycolysis and aerobic metabolism in the gills, aerobic metabolism in the liver and pentose-phosphate pathway in the kidneys during homeostasis. The biomarker response was tissue specific, with the greatest number of biomarkers altered in the gills, followed by those in the kidneys and liver.

Cold and warm waters: energy metabolism and antioxidant defenses of the freshwater fish Astyanax lacustris (Characiformes: Characidae) under thermal stress.

Subtropical fish are exposed to seasonal variations in temperature that impose a set of adaptations on their metabolism necessary for the maintenance of homeostasis. In this study, we addressed the effects of temperature variation on the metabolism of Astyanax lacustris, a species of freshwater fish common in the subtropical region of Brazil. Biomarkers of carbohydrate and protein metabolism, antioxidant defense, and oxidative damage were evaluated in the liver of A. lacustris exposed to low (15 °C) and high (31 °C) temperature thermal shock, with controls at 23 °C for 2, 6, 12, 24, 48, 72, and 96 h. A high energy demand was observed during the first 48 h of exposure to 15 °C, which is necessary for metabolic adjustment at low temperatures, with an increase in glycolysis, citric acid cycle, and amino acid catabolism. In addition, at 31 °C, glucose was exported in the first 12 h of exposure, and an increase in the citric acid cycle suggested acetyl-CoA as the pathway substrate, originating from the oxidation of lipids. The antioxidant defenses did not change at 15 °C, as opposed to 31 °C, in which there were changes in several antioxidant defense markers, indicating a response to the production of ROS. However, oxidative stress was observed at both temperatures, with oxidative damage detected by lipid peroxidation at 15 °C and protein carbonylation at 31 °C.

Dietary supplementation with nerolidol improves the antioxidant capacity and muscle fatty acid profile of Brycon amazonicus exposed to acute heat stress.

An increase in water temperature in the Amazon River has elicited concerns about commercially important fish species associated with food security, such as matrinxã (Brycon amazonicus). Studies have demonstrated the positive effects of diets supplemented with plant-based products that combat heat stress-induced oxidative damage. The aim of this study was to determine whether dietary supplementation with nerolidol prevents or reduces muscle oxidative damage and impairment of the fillet fatty acid profile of matrinxã exposed to heat stress. Plasma and muscle reactive oxygen species (ROS) and lipid peroxidation (LPO) levels were significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while plasma superoxide dismutase (SOD) and glutathione peroxidase (GPx) activity was significantly lower. The total content of saturated fatty acids (SFA) in fillets was significantly higher in fish exposed to heat stress compared to fish not exposed to heat stress, while he total content of polyunsaturated fatty acids (PUFA) was significantly lower. Nerolidol prevented the increase of muscle LPO and plasma ROS and LPO levels in fish exposed to heat stress, and partially prevented the increase in muscle ROS levels. Diets containing nerolidol prevented the inhibition of muscle GPx activity in fish exposed to heat stress, and partially prevented the decrease of plasma GPx activity. The nerolidol-supplemented diet prevented the increase of fillet SFA in fish exposed to heat stress, while partially preventing the decrease of PUFA. We conclude that acute heat stress at 34 °C for 72 h causes plasma and muscular oxidative damage, and that homeoviscous adaptation to maintain membrane fluidity can represent a negative impact for fish consumers. A nerolidol diet can be considered a strategy to prevent heat stress-induced oxidative damage and impairment of muscle fatty acid profiles.

Ecotoxicity of losartan potassium in aquatic organisms of different trophic levels.

The intensive use of the antihypertensive losartan potassium (LOS) has culminated in its high occurrence in aquatic environments. However, insufficient studies had investigated its effects in non-target organisms. In this study, ecotoxicity of LOS was assessed in aquatic organisms from distinct trophic levels (Desmodesmus subspicatus, Daphnia magna, and Astyanax altiparanae). Genotoxicity was assessed by the comet assay in D. magna and A. altiparanae, and biochemical biomarkers for the fish. LOS was more toxic to D. subspicatus (EC50(72h) = 27.93 mg L-1) than D. magna (EC50 = 303.69 mg L-1). Subsequently, this drug showed to induce more DNA damage in D. magna than A. altiparanae, when exposed to 2.5 mg L-1. No significant stress responses were observed by the fish biomarkers, suggesting that higher trophic levels organisms are more tolerant to LOS toxicity. LOS showed relatively low toxic potential for a short period of exposure, but with different patterns of toxicity for the organisms from distinct trophic levels, contributing to further risk assessment of LOS.

Silencing of Transposable Elements Mediated by 5-mC and Compensation of the Heterochromatin Content by Presence of B Chromosomes in Astyanax scabripinnis.

The way in which transcriptional activity overcomes the physical DNA structure and gene regulation mechanisms involves complex processes that are not yet fully understood. Modifications in the cytosine-guanine sequence of DNA by 5-mC are preferentially located in heterochromatic regions and are related to gene silencing. Herein, we investigate evidence of epigenetic regulation related to the B chromosome model and transposable elements in A. scabripinnis. Indirect immunofluorescence using anti-5-mC to mark methylated regions was employed along with quantitative ELISA to determine the total genomic DNA methylation level. 5-mC signals were dispersed in the chromosomes of both females and males, with preferential accumulation in the B chromosome. In addition to the heterochromatic methylated regions, our results suggest that methylation is associated with transposable elements (LINE and Tc1-Mariner). Heterochromatin content was measured based on the C-band length in relation to the size of chromosome 1. The B chromosome in A. scabripinnis comprises heterochromatin located in the pericentromeric region of both arms of this isochromosome. In this context, individuals with B chromosomes should have an increased heterochromatin content when compared to individuals that do not. Although, both heterochromatin content and genome methylation showed no significant differences between sexes or in relation to the occurrence of B chromosomes. Our evidence suggests that the B chromosome can have a compensation effect on the heterochromatin content and that methylation possibly operates to silence TEs in A. scabripinnis. This represents a sui generis compensation and gene activity buffering mechanism.

Sublethal effects of propiconazole on the metabolism of lambari Deuterodon iguape (Eigenmann 1907), a native species from Brazil.

The objective of this study was to analyze the sublethal effects of propiconazole on Deuterodon iguape, a native fish common in Brazil, which has potential for aquaculture and use as a bioindicator. The hypothesis was to test whether D. iguape has a metabolism similar to Danio rerio so that its use in bioassays may be validated. Lethal concentration (LC50) and metabolic rates were studied in fish exposed to propiconazole. Specific oxygen consumption and ammonia excretion for D. iguape and D. rerio increased by 0.01 µg L-1 and then decreased as the propiconazole concentration increased. The decrease in the averages of specific oxygen consumption at the concentration of 0.1 µg L-1 represented a reduction in the metabolic rate compared to the control of 71% for D. iguape and 40% D. rerio. For the ammonia excretion, at the same concentration, there was a reduction of 68.7% and 45.4% for D. iguape and D. rerio, respectively. When comparing ammonia excretion of the two species for each concentration of propiconazole, there was a significant difference (p < 0.05) in relation to the control and for the highest concentration (0.1 µg L-1). As for specific oxygen consumption, there was a statistically significant difference only for the concentration of 0.1 µg L-1. D. iguape proved to be a good and useful bioindicator for ichthyologists or ecologists in studies of moderate pesticide contamination in freshwater aquatic environments, as its metabolic response was similar to D. rerio.

Evaluation of the oestrogenic potential of oestrone and bisphenol-A on the reproduction of Astyanax bimaculatus males after subacute exposure.

In the last decades, oestrogenic compounds have often been reported in environmentally relevant concentrations in aquatic environments around the world. Most laboratory studies of oestrogens try to understand the effects of a single contaminant, but in natural environments, the effects may be quite different due to interactions with other compounds. The present study aimed to compare the action of oestrone (E1) and bisphenol-A (BPA), acting singularly and in combination, on the spermatogenesis of Astyanax bimaculatus. After exposure to 100 ng/L of E1, BPA and a mixture of the two for 15 days, our results showed that E1 and the E1 + BPA mixture significantly altered the number of spermatogenic cells. BPA presented high cytotoxicity when compared to other treatments. Analysis of the two oestrogenic compounds suggests that the E1 + BPA mixture has no additive or synergistic effects. Together, the results of the present study indicate that endocrine-disrupting chemicals (EDCs) analysed alone may behave differently than when administered with other substances.

Aluminum at environmental concentrations affects the sperm quality of the freshwater teleost Astyanax altiparanae: An in vitro study.

Aluminum (Al) is present in rivers and reservoirs in concentrations above that is allowed by regulatory agencies (e.g. 0.5 mg L-1 Al), which can impair fish reproduction. The present study evaluated the in vitro effects on the sperm of Astyanax altiparanae upon Al exposure at different concentrations (0, 0.05, 0.1, 0.3, and 0.5 mg L-1) with various exposure periods (50 s, 10 min, and 30 min). The following biomarkers were evaluated: membrane vitality, DNA fragmentation, morphology, kinetics (10 s and 30 s after sperm activation), and sperm mitochondrial activity. Al damages the membrane vitality of gametes at 0.3 and 0.5 mg L-1 after 50 s of exposure. After 30 min of exposure, there was a decrease in membrane vitality at 0.1 and 0.5 mg L-1, and the membrane vitality decreased with increased exposure time. Within 30 s after sperm activation, Al (0.3 and 0.5 mg L-1) reduced sperm motility by more than 50% at the longest exposure time, while at 0.1 and 0.5 mg L-1, Al exposure reduced motility over time. The average path speed (VAP; 10 s post-sperm activation) was reduced at longer exposure times at 0.05 and 0.5 mg L-1 of Al. Increased exposure time had deleterious effects on mitochondrial activity at the highest concentrations tested. Al did not damage DNA and sperm morphology. In conclusion, Al negatively influences the sperm quality of A. altiparanae with a potential effect of exposure time and increasing concentrations.

Effects of subchronic exposure to environmentally relevant concentrations of the herbicide atrazine in the Neotropical fish Astyanax altiparanae.

Atrazine (ATZ) is among the most widely used herbicides in the world, and yet it has a potential to contaminate aquatic environments due to pesticide leaching from agricultural areas. In the Neotropical region, studies about the effects of this herbicide in native aquatic wildlife is scarce.Our study aimed at investigating the effects of a 30-day exposure to a commercial atrazine formulation on oxidative stress parameters, histopathology in testis and liver, and hormone levels in males and female of yellow-tailed tetra fish (Astyanax altiparanae). Adults were exposed to low but environmentally relevant concentrations of atrazine as follows: 0 (CTL-control), 0.5 (ATZ0.5), 1 (ATZ1), 2 (ATZ2) and 10 (ATZ10) µg/L. Our results showed decreased GST activity in gills in all groups of exposed animals and increased CAT activity in gills from the ATZ10 group. In the liver, there was an increase in lipid peroxidation in fish from ATZ1 and ATZ2 groups. Histological analysis of the liver showed increased percentage of sinusoid capillaries in ATZ2 fish, increased vascular congestion in ATZ1 and increased leukocyte infiltration in the ATZ10 group. Hepatocyte diameter analysis revealed a decrease in cell size in all groups exposed to ATZ, and a decrease in hepatocyte nucleus diameter in ATZ1, ATZ2 and ATZ10 groups. Endocrine parameters did not show significant changes following ATZ exposure, although an increase of triiodothyronine/thyroxine (T3/T4) ratio was observed in ATZ2 fish. Our results provide evidence that even low, environmentally relevant concentrations of ATZ produced oxidative damage and histological alterations in adult yellow-tailed tetra.

Persistent growth of microtubules at low density.

Microtubules (MTs) often form a polarized array with minus ends anchored at the centrosome and plus ends extended toward the cell margins. Plus ends display behavior known as dynamic instability-transitions between rapid shortening and slow growth. It is known that dynamic instability is regulated locally to ensure entry of MTs into nascent areas of the cytoplasm, but details of this regulation remain largely unknown. Here, we test an alternative hypothesis for the local regulation of MT behavior. We used microsurgery to isolate a portion of peripheral cytoplasm from MTs growing from the centrosome, creating cytoplasmic areas locally depleted of MTs. We found that in sparsely populated areas MT plus ends persistently grew or paused but never shortened. In contrast, plus ends that entered regions of cytoplasm densely populated with MTs frequently transitioned to shortening. Persistent growth of MTs in sparsely populated areas could not be explained by a local increase in concentration of free tubulin subunits or elevation of Rac1 activity proposed to enhance MT growth at the cell leading edge during locomotion. These observations suggest the existence of a MT density-dependent mechanism regulating MT dynamics that determines dynamic instability of MTs in densely populated areas of the cytoplasm and persistent growth in sparsely populated areas.