[Sargassum seaweed assaults the French West Indies]. / Algues sargasses à l'assaut des Antilles.

SARGASSUM SEAWEED AS SAULTS THE FRENCH WEST INDIES. Since 2011, Martinique and the islands of Guadeloupe have been affected by repeated groundings, culminating in an exceptional wave in 2018. While the sargassum ( Sargassum natans and S. fluitans ) involved in these phenomena are neither toxic nor urticating, indirect toxicity linked to the presence of microorganisms and heavy metals (arsenic, mercury, etc.) in sargassum clusters has been described. Similarly, after a 24 to 48 hours stay on the shore, sargassum algae enter a putrefaction cycle responsible to produce hydrogen sulfide (H2S) and ammonia (NH3). The acute toxicity of these gases is well known. However, very few data are available on the clinical effects of prolonged exposure to low doses of H2S and NH3. Our team has recently described the syndromic features of chronic exposure, supposing for deleterious effects on the cardiovascular, respiratory and neurological systems.

ALGUES SARGASSES À L'ASSAUT DES ANTILLES. Depuis 2011, la Martinique et les îles de la Guadeloupe sont touchées par des échouements à répétition d'algues sargasses qui ont culminé avec une vague exceptionnelle en 2018. Si les sargasses (Sargassum natans et S. fluitans) impliquées dans ces phénomènes ne sont ni toxiques ni urticantes, une toxicité indirecte liée à la présence de micro-organismes et de métaux lourds (arsenic, mercure…) dans les amas de sargasses est décrite. De même, après un séjour de vingt-quatre à quarante-huit heures sur le littoral, les algues sargasses entrent dans un cycle de putréfaction responsable de la production d'hydrogène sulfuré (H2S) et d'ammoniac (NH3). La toxicité aiguë de ces gaz est bien connue. Il existe en revanche très peu de données disponibles sur les effets cliniques d'une exposition prolongée à de faibles doses d'H2S ou NH3. Notre équipe a récemment décrit le tableau syndromique de l'exposition chronique et suppose des effets délétères sur le système cardiovasculaire, respiratoire et neurologique.

Acute toxicity of total ammonia to Macrobrachium rosenbergii postlarvae at different salinity levels.

Nitrogen compounds, particularly ammonium, nitrite and nitrate, are a major problem in shrimp production systems. These compounds can accumulate in the aquatic environment and reach harmful or even lethal levels. Thus, monitoring the levels of nitrogenous compounds such as ammonia and studying their effects on the animals are essential. One tool used for this purpose is acute toxicity testing based on the evaluation of LC50 values. Furthermore, tools that can help improve the performance of aquatic organisms in culture are needed. The present study aimed to evaluate the effect of salinity on the toxicity of total ammonia to postlarvae of the freshwater prawn Macrobrachium rosenbergii. For this purpose, acute toxicity testing (LC50-96h) was performed using 540 postlarvae with a mean weight of 0.13 g and a mean total length of 2.47 cm, divided into 54 experimental units of two liters each. A completely randomized design in a 3×6 factorial scheme was used, combining three salinities (0, 5, and 10 g.L-1) and six total ammonia concentrations (0, 8, 16, 32, 64, and 128 mg.L-1), with three replicates per combination. The LC50 values for M. rosenbergii postlarvae at 24, 48, 72, and 96 h and their respective confidence intervals (95%) were estimated using the trimmed Spearman-Karber method. The results showed that salinities of 5 or 10 g.L-1 did not reduce the acute toxicity of total ammonia.

Lethal and Sublethal Toxicity of Un-Ionized Ammonia to Early-Life Stages of Danio rerio.

Un-ionized ammonia (NH3) is a prevalent contaminant found in aquatic ecosystems, frequently associated with complex mixtures of other toxic contaminants. Early embryo-larval stages of zebrafish Danio rerio became an important model for water quality monitoring, and it is necessary to characterize its susceptibility to NH3 exposure. Fertilized eggs were exposed to NH3 concentrations ranging from 0.02 to 5.23 mg NH3 L-1 until 168 h postfertilization (hpf). The lethal concentration to 50% of exposed zebrafish during 96 h was 2.07 mg NH3 L-1, 25% above the median value reported values for early developmental stages of fishes. Sublethal toxicity endpoints indicated the lowest observed effect concentrations for slow blood circulation at 24 hpf, decreased heart ventricular contractions at 72 hpf, incomplete yolk sac absorption at 96 hpf, failure in swim bladder inflation at 96 hpf, developmental retardation at 96 hpf, decreased total length, decreased swimming speed, altered trajectories, and acetylcholinesterase inhibition at 168 hpf of 0.85, 0.06, 0.15, 0.06, 0.15, 0.61, 1.35, 0.35, and 0.85 mg NH3 L-1, respectively. Environmentally relevant NH3 concentrations can affect zebrafish's early development and larval viability, and our results help discriminate NH3 contribution to the toxicity of complex environmental mixtures when zebrafish is used in water quality monitoring.

Copper II oxide nanoparticles (CuONPs) alter metabolic markers and swimming activity in zebra-fish (Danio rerio).

The present study aimed to compare the metabolic effects caused by using copper oxide nanoparticles with two distinct morphologies nanorods and nanosphere. The CuONPs in the form of nanorods were characterized in the order of 500 nm, on a scale of 20, 100, and 500 nm. Meanwhile, the nanosphere CuONPs were characterized in the order of 5 nm, on a 30 nm scale. The analysis of metabolic rate was performed using the closed respirometry technique, specific ammonia excretion, and swimming ability as biomarkers, the physiological effects on Danio rerio were investigated. For the experiments, 88 fish were used, exposed for 24 h at concentrations of 0, 50, 100, and 200 µg/L of copper oxide nanoparticles in the form of nanospheres and nanorods, respectively. The tests carried out with the nanorods demonstrated metabolic alterations in fish, with an increase of 294% and 321% in the metabolic rate at concentrations of 100 µg/L and 200 µg/L, respectively. Furthermore, there was a decrease in specific ammonia excretion by 34% and 83% and in swimming capacity by 34% and 55% at concentrations of 100 and 200 µg/L, respectively, when compared to the control. The tests performed with nanospheres did not show significant changes compared to the control. These experiments showed that different morphological structures of the same copper oxide nanoparticle caused different effects on fish metabolism. It is concluded that the characterization of nanoparticles is essential to understand their effects on fish, since their structural forms can cause different toxic effects on D. rerio.

Effects of ammonia on gill (Na+, K+)-ATPase kinetics in a hololimnetic population of the Amazon River shrimp Macrobrachium amazonicum.

Water quality is essential for successful aquaculture. For freshwater shrimp farming, ammonia concentrations can increase considerably, even when culture water is renewed frequently, consequently increasing the risk of ammonia intoxication. We investigated ammonia lethality (LC50-96 h) in a hololimnetic population of the Amazon River shrimp Macrobrachium amazonicum from the Paraná/Paraguay River basin, including the effects of exposure to 4.93 mg L-1 total ammonia concentration on gill (Na+, K+)-ATPase activity. The mean LC50-96 h was 49.27 mg L-1 total ammonia, corresponding to 1.8 mg L-1 un-ionized ammonia. Except for NH4+ affinity that increased 2.5-fold, that of the gill (Na+, K+)-ATPase for ATP, Mg2+, Na+, K+ and ouabain was unchanged after ammonia exposure. Western blotting of gill microsomal preparations from fresh caught shrimps showed a single immunoreactive band of ≈110 kDa, corresponding to the gill (Na+, K+)-ATPase α-subunit. Ammonia exposure increased (Na+, K+)-ATPase activity by ≈25%, coincident with an additional 130 kDa α-subunit immunoreactive band, and increased K+-stimulated and V(H+)-ATPase activities by ≈2.5-fold. Macrobrachium amazonicum from the Paraná/Paraguay River basin is as tolerant to ammonia as are other Amazon River basins populations, showing toxicity comparable to that of marine crustaceans.

Expression of ion transport proteins and routine metabolism in juveniles of tropical gar (Atractosteus tropicus) exposed to ammonia.

Tropical gar (Atractosteus tropicus) thrives in aquatic habitats with high levels of total nitrogen (TAN) and unionized ammonia (NH3). However, the tolerance of TAN and NH3, the excretion mechanisms involved, and the effects of these chemicals on routine metabolism are still unknown. Therefore, our objectives were to assess the acute toxicity of TAN and NH3 in A. tropicus juveniles after a 96-h exposure (LC50-96 h) to NH4Cl and after chronic exposure to two concentrations (15% and 30% of LC50-96 h TAN) for 12 days, as well as to evaluate the transcriptional effects associated with Rhesus proteins (rhag, rhbg, rhcg) and ion transporters (NHE, NKA, NKCC, and CFTR) in gills and skin; and to determine the effects of TAN and NH3 on routine metabolism through oxygen consumption (µM g-1 h-1) and gill ventilation frequency (beats min-1). LC50-96 h values were 100.20 ± 11.21 mg/L for TAN and 3.756 ± 0.259 mg/L for NH3. The genes encoding Rhesus proteins and ion transporters in gills and skin showed a differential expression according to TAN concentrations and exposure time. Oxygen consumption on day 12 showed significant differences between treatments with 15% and 30% TAN. Gill ventilation frequency on day 12 was higher in fish exposed to 30% TAN. In conclusion, A. tropicus juveniles are highly tolerant to TAN, showing upregulation of the genes involved in TAN excretion through gills and skin, which affects routine oxygen consumption and energetic cost. These findings are relevant for understanding adaptations in the physiological response of a tropical ancestral air-breathing fish.

Protective effect of resveratrol on citrullinemia type I-induced brain oxidative damage in male rats.

Citrullinemia Type I is an inborn error, which leads to accumulation of citrulline and ammonia in blood and body tissues. We evaluated the in vitro effects of citrulline, ammonia and the influence of resveratrol on oxidative stress parameters in the cerebrum of 30- and 60-day-old male Wistar rats. Citrulline (0.1, 2.5, 5.0 mM), ammonia (0.01, 0.1, 1.0 mM) and resveratrol (0.01, 0.1, 0.5 mM) were added to the assays to measure thiobarbituric acid reactive substances (TBA-RS), total sulfhydryl content and the activity of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). Citrulline (2.5 and 5.0 mM) increased TBA-RS in the cerebellum of 30-day-old and in the cerebral cortex and cerebellum of 60-day-old. Citrulline (5.0 mM) increased SOD and reduced GSH-Px in the hippocampus of 30-day-old, whereas in the cerebellum it increased GSH-Px. In the cerebral cortex, 2.5 and 5.0 mM citrulline reduced GSH-Px. In 60-day-old, 2.5 and 5.0 mM citrulline increased SOD in the cerebellum, increased GSH-Px in the cerebral cortex and 5.0 mM citrulline reduced CAT and increased SOD in the cerebral cortex. Ammonia (0.1 and 1.0 mM) reduced the sulfhydryl content in the cerebral cortex of 30- and 60-day-old, 1.0 mM ammonia increased SOD and reduced GSH-Px in the cerebellum of 30-day-old and increased SOD in the hippocampus and cerebellum of 60-day-old. Resveratrol was able to prevent the majority of these alterations. Thus, citrulline and ammonia induce oxidative stress in the cerebrum of rats; however, resveratrol was able to exert antioxidant effects against these substances.

Effects of resveratrol on alterations in cerebrum energy metabolism caused by metabolites accumulated in type I citrullinemia in rats.

We investigated the in vitro effects of citrulline (0.1, 2.5 and 5.0 mM) and ammonia (0.01, 0.1 and 1.0 mM), and the influence of resveratrol (0.01 mM, 0.1 mM and 0.5 mM) on pyruvate kinase, citrate synthase, succinate dehydrogenase (SDH), complex II, and cytochrome c oxidase activities in cerebral cortex, cerebellum and hippocampus homogenates of 60-day-old male Wistar rats. Results showed that 2.5 and 5.0 mM citrulline decreased pyruvate kinase activity in cerebral cortex and, at a concentration of 5.0 mM, increased its activity in hippocampus. Additionally, 5.0 mM citrulline increased citrate synthase activity in the cerebellum of rats. Citrulline (5.0 mM) reduced complex II and cytochrome c oxidase activities in cerebral cortex and hippocampus. With regard to ammonia, at 0.1 and 1.0 mM, decreased complex II activity in cerebral cortex and at 1.0 mM decreased its activity in cerebellum and hippocampus. Ammonia (1.0 mM) also decreased cytochrome c oxidase activity in cerebral cortex and cerebellum of rats. Resveratrol was able to prevent most of the alterations caused by these metabolites in the biomarkers of energy metabolism measured in the cerebrum of rats. Data suggest that these alterations in energy metabolism, caused by citrulline and ammonia, are probably mediated by the generation of free radicals, which can in turn be scavenged by resveratrol.

Ammonia exposition during gestation induces neonatal oxidative damage in the brain and long-term cognitive alteration in rats.

Ammonia is involved in the pathogenesis of neurological conditions associated with hyperammonemia, including hepatic encephalopathy. Few is known about the effects of gestational exposition to ammonia in the developing brain, and the possible long-term consequences of such exposure. We aimed to evaluate the effects of ammonia exposure during the gestation and the possible long-term cognitive alterations on pups. Eight female rats were divided into two groups: (1) control (saline solution); (2) ammonia (ammonium acetate, 2,5mmol/Kg). Each rat received a single subcutaneous injection during all gestational period. The brains from 1-day-old rats were obtained to the determination of thiobarbituric acid reactive species (TBARS), protein carbonyl and nitrite/nitrate levels. Some animals were followed 30 days after delivery and were subjected to the step-down inhibitory avoidance task. It was observed a significant increase in protein carbonyl, but not TBARS or nitrite/nitrate levels, in pups exposed to ammonia. Rats exposed to ammonia presented long-term cognitive impairment. Gestational exposition to ammonia induces protein oxidative damage in the neonatal rat brain, and long-term cognitive impairment.

Toxicity of ammonia, nitrite and nitrate to Litopenaeus vannamei juveniles in low-salinity water in single and ternary exposure experiments and their environmental implications.

Information on toxicity of nitrogen compounds for Litopenaeus vannamei in coastal ecosystems and culture under low salinity is scarce. Acute toxicity trials were conducted in L. vannamei to determine the single and combined effects of ammonia, nitrite and nitrate at a salinity of 3 g/L. The 96 h-LC50 was 29.0 mg/L for total ammonia nitrogen (TAN); 10.6 mg/L for nitrogen as nitrite (NO2--N); and 900 mg/L for nitrogen as nitrate (NO3--N). The joint effects of ammonia, nitrite and nitrate exposure were antagonistic at 24-72 h; and additive from 72 to 96 h. The proposed safety levels of single exposure to TAN, NO2--N and NO3--N for L. vannamei are 1.45, 0.53 and 45.0 mg/L, respectively. When in mixture, the proposed level of TAN/NO2--N/NO3--N is 0.05 TU (Toxicity Unit) corresponding to 0.48, 0.08 and 14.6 mg/L of TAN, NO2--N and NO3--N, respectively.

Multiple lines of evidence of sediment quality in an urban Marine Protected Area (Xixová-Japuí State Park, SP, Brazil).

Marine Protected Areas (MPAs) aim to protect habitats, biodiversity, and ecological processes as a conservation tool. These areas have been affected by contamination, which threats the biodiversity and ecological functioning. In this study, we evaluated the sediment quality of Xixová-Japuí State Park (XJSP), an MPA located in an urbanized Bay (Santos, Southeast Brazil) by integrating multiple lines-of-evidence. Six sites were selected within the XJSP and analyzed for sediment chemistry, toxicity, and benthic community descriptors using Sediment Quality Triad approach (SQT). Whole-sediment Toxicity Identification Evaluation (TIE) was employed as a complementary line of evidence to confirm the presence of domestic effluent discharges as a potential stressor. The SQT showed that sediments collected within XJSP are impacted by contaminants, exhibiting chronic toxicity and changes in benthic community. TIE results indicated that trace metals, organic contaminants, and ammonia contributed to the observed effects. Our results also indicate a lack of effectiveness of MPA in protecting the biodiversity due to the contamination sources, which requires efforts to pollution control in order to ensure the environmental conservation and management plan goals.

Flesh quality and stress responses of Piaractus mesopotamicus after exposure to sublethal levels of ammonia and subsequent recovery / Qualidade da carne e respostas de estresse em Piaractus mesopotamicus após exposição à níveis subletais de amônia e subsequente recuperação

This study aimed to evaluate de effect of the exposure to sub-lethal levels of ammonia on blood parameters and flesh quality of juvenile pacu (Piaractus mesopotamicus), including assessment of such parameters post recovery in ammonia-free water. Juveniles (27.1 ± 5.4 g) were exposed to concentrations of ammonia at 0.0 (control); 0.5; and 1.0 mg N-NH3 L-1 for 10 days followed by the same period of recovery in ammonia-free water. On the 10th day post exposure and after a recovery period, samples of blood were taken for glucose, lactate analyses and evaluation of haematocrit. To evaluate lipid peroxidation, proximate composition and sensory analysis, samples of muscle/fillets were also obtained. Exposure to ammonia caused alterations in haematological response and negatively affected sensory analysis of pacu fillet. However, proximate composition was unchanged and lipid peroxidation process was not intensified in muscle. In conclusion, exposure to sublethal levels of ammonia induces secondary stress responses and altered the organoleptic characteristics of pacu flesh. Nevertheless, a recovery period of 10 days was sufficient to allow for a complete restoration of the homeostasis and organoleptic characteristics of the fillet.

O objetivo do presente estudo foi avaliar os efeitos da exposição a níveis subletais de amônia nos parâmetros sanguíneos e qualidade da carne de juvenis de pacu (Piaractus mesopotamicus). Além disso, também foi realizada a avaliação desses parâmetros pós-recuperação em água livre de amônia. Os peixes (27,1 ± 5,4 g) foram expostos a 0,0 (controle); 0,5 e 1,0 mg N-NH3 L-1 por 10 dias seguidos pelo mesmo tempo de recuperação em água livre de amônia. Ao final dos 10 dias de exposição e após o período de recuperação foram coletadas amostras de sangue para determinação dos níveis de glicose, lactato e dos valores de hematócrito. Para avaliação dos níveis de peroxidação lipídica, da composição proximal e realização da análise sensorial, amostras de músculo/filé também foram coletadas. A exposição à amônia ocasionou alterações nos parâmetros sanguíneos e afetou negativamente a análise sensorial dos filés de pacu. Entretanto, a composição proximal e o grau de peroxidação lipídica não foram alterados. Concluindo, a exposição a níveis subletais de amônia induziu respostas secundárias de estresse em juvenis de pacu e alterou características organolépticas do filé. Um período de recuperação de 10 dias foi suficiente para restaurar a homeostase e a qualidade do filé.

Lethal and sublethal effects of ammonia in Deuterodon iguape (Eigenmann 1907), potential species for Brazilian aquaculture / Efeito letal e subletal da amônia sobre o lambari (Deuterodon iguape, Eigenmann 1907), espécie potencial para a aquicultura brasileira

The cultivation of the lambari, Deuterodon iguape, supplies much of the Brazilian market with live bait for sport fishing. The high densities used to maximize production can increase the concentration of ammonia. In order to evaluate the sublethal and lethal effects of different concentrations of ammoniacal nitrogen (non-ionized ammonia plus ionized ammonia), D. iguape were exposed to this xenobiotic. The LC50 values for 24, 48, 72, 96 h of ammonia-N were 6.17, 5.57, 3.88 and 2.90 mg L-1 at 23°C. The LC50 values of 24, 48, 72, 96 h of NH3-N (non-ionized ammonia with nitrogen) were 0.015; 0.013; 0.009; 0.007 mg L-1. The specific oxygen consumption increased at the ammonia-N concentrations tested. The values for the concentrations of 0.1; 0.25; 0.5 and 1.0 mg L-1 were: 0.25, 0.33; 0.31 and 0.44 mlO2 g-1 h-1. At the concentration of 1.0 mg L-1 of ammonium chloride there was a 41.33% increase in the consumption level in relation to the control. Ammonia excretion also increased with increasing ammoniacal nitrogen concentration. The fish excreted, on average, 0.0320, 0.0364, 0.0368 and 0.0370 mg/g/h of ammonia. Comparing these results with the means of ammonia excretion of the control (0.0 mg L-1), it was observed that these values represent an increase in the excretion of 146%, 177%, 189% and 184%, respectively. After 24 h of exposure to ammonia, the O:N ratio decreased by 43.46%. Our results indicate pronounced metabolic effects and increased toxicity with increasing ammonia concentrations. We recommend avoiding concentrations higher than 0.25 mg L-1 NH3-N in the culture water.

O cultivo do lambari, Deuterodon iguape, abastece grande parte do mercado brasileiro com iscas vivas para a pesca esportiva. As altas densidades utilizadas para maximizar a produção podem aumentar a concentração de amônia. A fim de avaliar os efeitos subletais e letais de diferentes concentrações de nitrogênio amoniacal (amônia não ionizada mais amônia ionizada), D. iguape foram expostos a este xenobiótico. Os valores de CL50 para 24, 48, 72, 96 h de amônia-N foram 6,17; 5,57; 3,88 e 2,90 mg L-1 a 23 °C. Os valores de CL50 de 24, 48, 72, 96 h de NH3-N (amónia não ionizada) foram de 0,015; 0,013; 0,009 e 0,007 mg L-1. O consumo específico de oxigênio aumentou nas concentrações de nitrogênio amoniacal testadas. Os valores para as concentrações de 0,1; 0,25; 0,5 e 1,0 mg L-1 foram: 0,25, 0,33; 0,31 e 0,44 mlO2 g-1 h-1. Na concentração de 1,0 mg L-1 de cloreto de amônio houve aumento de 41,33% no nível de consumo em relação ao controle. A excreção de amônia também aumentou com o aumento da concentração de nitrogênio amoniacal. Os peixes excretaram, em média, 0,0320; 0,0364; 0,0368 e 0,0370 mg/g/h de amônia. Comparando esses resultados com as médias de excreção de amônia do controle (0,0 mg L-1), observou-se que esses valores representam um aumento na excreção de 146%, 177%, 189% e 184%, respectivamente. Após 24 h de exposição à amônia, a relação O: N diminuiu em 43,46%. Nossos resultados indicam efeitos metabólicos pronunciados e aumento da toxicidade com o aumento das concentrações de amônia. Recomendamos evitar concentrações superiores a 0,25 mg L-1 de NH3-N na água da cultura para o D. iguape.

Lethal and sublethal effects of ammonia in Deuterodon iguape (Eigenmann 1907), potential species for Brazilian aquaculture / Efeito letal e subletal da amônia sobre o lambari (Deuterodon iguape, Eigenmann 1907), espécie potencial para a aquicultura brasileira

The cultivation of the lambari, Deuterodon iguape, supplies much of the Brazilian market with live bait for sport fishing. The high densities used to maximize production can increase the concentration of ammonia. In order to evaluate the sublethal and lethal effects of different concentrations of ammoniacal nitrogen (non-ionized ammonia plus ionized ammonia), D. iguape were exposed to this xenobiotic. The LC50 values for 24, 48, 72, 96 h of ammonia-N were 6.17, 5.57, 3.88 and 2.90 mg L-1 at 23°C. The LC50 values of 24, 48, 72, 96 h of NH3-N (non-ionized ammonia with nitrogen) were 0.015; 0.013; 0.009; 0.007 mg L-1. The specific oxygen consumption increased at the ammonia-N concentrations tested. The values for the concentrations of 0.1; 0.25; 0.5 and 1.0 mg L-1 were: 0.25, 0.33; 0.31 and 0.44 mlO2 g-1 h-1. At the concentration of 1.0 mg L-1 of ammonium chloride there was a 41.33% increase in the consumption level in relation to the control. Ammonia excretion also increased with increasing ammoniacal nitrogen concentration. The fish excreted, on average, 0.0320, 0.0364, 0.0368 and 0.0370 mg/g/h of ammonia. Comparing these results with the means of ammonia excretion of the control (0.0 mg L-1), it was observed that these values represent an increase in the excretion of 146%, 177%, 189% and 184%, respectively. After 24 h of exposure to ammonia, the O:N ratio decreased by 43.46%. Our results indicate pronounced metabolic effects and increased toxicity with increasing ammonia concentrations. We recommend avoiding concentrations higher than 0.25 mg L-1 NH3-N in the culture water.(AU)

O cultivo do lambari, Deuterodon iguape, abastece grande parte do mercado brasileiro com iscas vivas para a pesca esportiva. As altas densidades utilizadas para maximizar a produção podem aumentar a concentração de amônia. A fim de avaliar os efeitos subletais e letais de diferentes concentrações de nitrogênio amoniacal (amônia não ionizada mais amônia ionizada), D. iguape foram expostos a este xenobiótico. Os valores de CL50 para 24, 48, 72, 96 h de amônia-N foram 6,17; 5,57; 3,88 e 2,90 mg L-1 a 23 °C. Os valores de CL50 de 24, 48, 72, 96 h de NH3-N (amónia não ionizada) foram de 0,015; 0,013; 0,009 e 0,007 mg L-1. O consumo específico de oxigênio aumentou nas concentrações de nitrogênio amoniacal testadas. Os valores para as concentrações de 0,1; 0,25; 0,5 e 1,0 mg L-1 foram: 0,25, 0,33; 0,31 e 0,44 mlO2 g-1 h-1. Na concentração de 1,0 mg L-1 de cloreto de amônio houve aumento de 41,33% no nível de consumo em relação ao controle. A excreção de amônia também aumentou com o aumento da concentração de nitrogênio amoniacal. Os peixes excretaram, em média, 0,0320; 0,0364; 0,0368 e 0,0370 mg/g/h de amônia. Comparando esses resultados com as médias de excreção de amônia do controle (0,0 mg L-1), observou-se que esses valores representam um aumento na excreção de 146%, 177%, 189% e 184%, respectivamente. Após 24 h de exposição à amônia, a relação O: N diminuiu em 43,46%. Nossos resultados indicam efeitos metabólicos pronunciados e aumento da toxicidade com o aumento das concentrações de amônia. Recomendamos evitar concentrações superiores a 0,25 mg L-1 de NH3-N na água da cultura para o D. iguape.(AU)

Flesh quality and stress responses of Piaractus mesopotamicus after exposure to sublethal levels of ammonia and subsequent recovery / Qualidade da carne e respostas de estresse em Piaractus mesopotamicus após exposição à níveis subletais de amônia e subsequente recuperação

This study aimed to evaluate de effect of the exposure to sub-lethal levels of ammonia on blood parameters and flesh quality of juvenile pacu (Piaractus mesopotamicus), including assessment of such parameters post recovery in ammonia-free water. Juveniles (27.1 ± 5.4 g) were exposed to concentrations of ammonia at 0.0 (control); 0.5; and 1.0 mg N-NH3 L-1 for 10 days followed by the same period of recovery in ammonia-free water. On the 10th day post exposure and after a recovery period, samples of blood were taken for glucose, lactate analyses and evaluation of haematocrit. To evaluate lipid peroxidation, proximate composition and sensory analysis, samples of muscle/fillets were also obtained. Exposure to ammonia caused alterations in haematological response and negatively affected sensory analysis of pacu fillet. However, proximate composition was unchanged and lipid peroxidation process was not intensified in muscle. In conclusion, exposure to sublethal levels of ammonia induces secondary stress responses and altered the organoleptic characteristics of pacu flesh. Nevertheless, a recovery period of 10 days was sufficient to allow for a complete restoration of the homeostasis and organoleptic characteristics of the fillet.(AU)

O objetivo do presente estudo foi avaliar os efeitos da exposição a níveis subletais de amônia nos parâmetros sanguíneos e qualidade da carne de juvenis de pacu (Piaractus mesopotamicus). Além disso, também foi realizada a avaliação desses parâmetros pós-recuperação em água livre de amônia. Os peixes (27,1 ± 5,4 g) foram expostos a 0,0 (controle); 0,5 e 1,0 mg N-NH3 L-1 por 10 dias seguidos pelo mesmo tempo de recuperação em água livre de amônia. Ao final dos 10 dias de exposição e após o período de recuperação foram coletadas amostras de sangue para determinação dos níveis de glicose, lactato e dos valores de hematócrito. Para avaliação dos níveis de peroxidação lipídica, da composição proximal e realização da análise sensorial, amostras de músculo/filé também foram coletadas. A exposição à amônia ocasionou alterações nos parâmetros sanguíneos e afetou negativamente a análise sensorial dos filés de pacu. Entretanto, a composição proximal e o grau de peroxidação lipídica não foram alterados. Concluindo, a exposição a níveis subletais de amônia induziu respostas secundárias de estresse em juvenis de pacu e alterou características organolépticas do filé. Um período de recuperação de 10 dias foi suficiente para restaurar a homeostase e a qualidade do filé.(AU)

Acute Toxicity of Ammonia, Nitrite and Nitrate to Shrimp Litopenaeus vannamei Postlarvae in Low-Salinity Water.

Shrimp farming in low salinities waters is an alternative to increasing production, and counteracting disease problems in brackish and marine waters. However, in low-salinity waters, toxicity of nitrogen compounds increases, and there is no available data of its acute toxicity in shrimp postlarvae. This study determined the acute toxicity of ammonia, nitrite and nitrate in Litopenaeus vannamei postlarvae in 1 and 3 g/L salinity, as well as the safety levels. The LC50 confirms that nitrite is more toxic than ammonia and nitrate in low salinity waters, and that its toxicity increases with a decrease in salinity. The safe levels estimated for salinities of 1 and 3 g/L were 0.54 and 0.81 mg/L for total ammonia-N, 0.17 and 0.25 mg/L for NO2-N, and 5.6 and 21.5 mg/L for NO3-N, respectively.

Biological responses of Neotropical freshwater fish Lophiosilurus alexandri exposed to ammonia and nitrite.

This study aimed to elucidate the responses of the Neotropical fish Lophiosilurus alexandri exposed to ammonia and nitrite, following a period of recovering. Acute toxicity tests lasted 96h, subchronic toxicity tests lasted eight days and the detoxification trial lasted four days. Groups of 12 juveniles were maintained in 90-L tanks and treated with increasing concentrations of ammonia and nitrite, except during the recovery test. All treatments were performed with two replicates. The median lethal concentrations (LC50) of 24, 48, 72 and 96h were estimated at 30.12; 24.35; 19.24 and 18.68mg·L-1 TA-N; 5.37; 4.57; 3.75 and 3.66mg·L-1 NH3-N and 20.37; 7.78; 7.09 and 5.86mg·L-1 NO2--N, respectively. The NO2- caused significant decrease in hematocrit and increase in the urea levels during short-term exposure, with recovery of homeostasis after the subchronic and detox period. Acute exposure to ammonia increased the enzyme profile of transaminases, glucose and urea. Urea concentration remained high in the subchronic and detox tests. Histopathologies were observed in animals exposed to ammonia in both toxicity tests. It was highlighted detachment of epithelium, hyperemia and necrosis in the gills. Exposure to NO2- caused epithelium detachment and aneurysm. Vacuolization and swelling of hepatocytes were the most common injury for both nitrogenous compounds. We concluded that the L. alexandri has moderate tolerance to ammonia and nitrite. The recovery period revealed remedial response to ammonia and nitrite exposure.

Resveratrol prevents ammonia-induced mitochondrial dysfunction and cellular redox imbalance in C6 astroglial cells.

BACKGROUND: Resveratrol is a polyphenolic compound that presents several protective effects in the central nervous system, including gliotoxicity associated to hyperammonemia, a key element for the development of hepatic encephalopathy. In this condition, mitochondrial dysfunction leads to a reactive oxygen species (ROS) overproduction, which, in turn, exacerbates mitochondrial failure and causes cellular damage. OBJECTIVE: This study sought to determine whether prevention of mitochondrial dysfunction and the maintenance of cellular redox status by resveratrol contribute to its protective action toward ammonia toxicity. METHODS: C6 astrocyte cell line was pre-incubated in the presence or absence of resveratrol (100 µM) for 1 hour. After pre-incubation, resveratrol was maintained and 5 mM ammonia was added for 24 hours, followed by the evaluation of ROS production, mitochondrial functionality, antioxidant enzymatic and non-enzymatic defenses, energy metabolic parameters, and genotoxicity. RESULTS: We showed that resveratrol prevented the increase in ROS production, the decrease of mitochondrial membrane potential (ΔΨm), and bioenergetics deficit caused by ammonia in C6 astroglial cells. In addition, resveratrol avoided the ammonia-induced upregulation of NOX activity and impairment in enzymatic and non-enzymatic antioxidant defenses. Ammonia also induced DNA damage that was prevented by resveratrol, indicating its genoprotective effect. CONCLUSIONS: In summary, our study demonstrates that resveratrol prevents ammonia-induced cytotoxicity, as well as supports the role of resveratrol on mitochondrial/cellular redox functionality.

Ammonia exposure and subsequent recovery trigger oxidative stress responses in juveniles of Brazilian flounder Paralichthys orbignyanus.

The effects of ammonia exposure and recovery on oxidative stress parameters and histology of juvenile Brazilian flounder Paralichthys orbignyanus were evaluated. The fish were exposed to 0.12, 0.28 and 0.57 mg NH3-N L-1, plus a control, for 10 days followed by the same recovery time in ammonia-free water. Gill, liver and muscle samples (n = 9) were collected after 1, 5 and 10 days of exposure and after recovery for oxidative stress analysis (antioxidant capacity against peroxyl radicals (ACAP); glutathione S-transferase (GST) activity; lipoperoxidation levels measured through thiobarbituric acid reactive substances (TBARS) content). For histological assessment, gill, liver and brain samples were collected. Exposure to all NH3-N concentrations induced different time- and dose-dependent changes in oxidative stress parameters. Reduced antioxidant capacity of the liver and muscle and enhanced TBARS levels in the gills and liver were demonstrated. Differently, a high ammonia concentration elicited lower hepatic TBARS levels. Enhanced GST activity in all organs and increased antioxidant capacity of the gills were also observed. No ammonia-induced histopathological effects were demonstrated. After recovery, most parameters (liver ACAP, GST activity in the muscle and liver and TBARS in the gills) returned to baseline levels. However, liver TBARS and gill GST activity remained altered 0.57 mg NH3-N L-1 treatment. The recovery period also led to a decrease in gill antioxidant capacity and an increase in muscle antioxidant capacity. In conclusion, a concentration of 0.12 mg NH3-N L-1 induces oxidative stress and antioxidant responses in juvenile Brazilian flounder. Moreover, a 10-day recovery period is not sufficient to restore fish homeostasis.

Chronic effects of nitrogenous compounds on survival and growth of juvenile pink shrimp / Efeito crônico de compostos nitrogenados sobre a sobrevivência e crescimento de juvenis de camarão rosa

Abstract In response to growing worldwide market demand, intensive shrimp farming, based on high feed, has developed over the past decade. The nitrogenous compounds mainly generated by animal excretion can cause deterioration of water quality and produce chronic or even acute toxicity to aquatic animals. As prevention, theoretical safety levels have been estimated from acute toxicity tests and they are traditionally used to prevent toxic effects on biota. However, are those concentrations of nitrogenous compounds really safe to Farfantepenaeus paulensis? The current study aimed to investigate the lethal and sublethal effects of ammonia, nitrite and nitrate to juvenile F. paulensis based on safety levels. Each experiment was performed independently in 100 L tanks for 30 days. The survival rates and wet weight of all shrimps were recorded every 10 days. The concentrations tested for ammonia, nitrite and nitrate were respectively: treatment "T1/4", a quarter of the safety level (0.91 mg/L TA-N, 2.55 mg/L NO2--N and 80.7 mg/L NO3--N); treatment "TSL", the safety level (3.65 mg/L TA-N, 10.2 mg/L NO2--N and 323 mg/L NO3--N); and treatment "T2X", twice the safety level (7.30 mg/L TA-N, 20.4 mg/L NO2--N and 646 mg/L NO3--N). For F. paulensis cultivation, the real safety level for nitrite was estimated to be 2.55 mg/L NO2--N. For ammonia and nitrate, the recommended concentrations were <0.91 mg/L TA-N corresponding to 0.045 mg/L NH3-N and <80.7 mg/L NO3--N, respectively.

Resumo Em resposta à crescente demanda do mercado mundial, a carcinicultura intensiva tem se desenvolvido ao longo da última década. Os compostos nitrogenados gerados principalmente pela excreção dos animais podem causar a deterioração da qualidade da água e produzir toxicidade crônica ou mesmo aguda para os animais cultivados. Como prevenção, os níveis de segurança teóricos são estimados a partir de testes de toxicidade aguda e são tradicionalmente usados para evitar efeitos tóxicos sobre a biota. No entanto, as estimativas das concentrações dos compostos nitrogenados são realmente seguras para Farfantepenaeus paulensis? O presente estudo teve como objetivo investigar os efeitos letais e subletais da amônia, nitrito e nitrato em juvenis de camarão marinho F. paulensis com base em níveis de segurança. Cada experimento foi realizado de forma independente em tanques com capacidade de 100 L durante 30 dias. As taxas de sobrevivência e peso úmido de todos os camarões foram registrados a cada 10 dias. As concentrações testadas para amônia, nitrito e nitrato foram respectivamente: "T1/4", um quarto do nível de segurança (0,91 mg/L N-AT, 2,55 mg/L de N-NO2- e 80,7 mg/L N-NO3-); "TSL", nível de segurança (3,65 mg/L N-AT, 10,2 mg/L de N-NO2- e 323 mg/L N-NO3-); e "T2X", duas vezes o nível de segurança (7,30 mg/L N-AT, 20,4 mg/L de N-NO2- e 646 mg/L de N-NO3-). Para a criação de F. paulensis, o nível de segurança real para nitrito foi estimado em 2,55 mg/L N-NO2-. Para amônia e nitrato, concentrações recomendadas foram: <0,91 mg/L N-AT correspondente a 0,045 mg/L N-NH3 e <80,7 mg/L N-NO3-, respectivamente.