A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals / Comportamento fisiológico e tolerância de Lactuca sativa aos metais pesados nitrato de chumbo e nitrato de prata

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-1, Pb = 180 mg. Kg-1), double (Ag = 50 mg. Kg-1, Pb = 360 mg. Kg-1) and triple (Ag = 75 mg. Kg-1, Pb = 540 mg. Kg -1) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Este estudo teve como objetivo avaliar os efeitos de diferentes concentrações de chumbo (Pb) e prata (Ag) na germinação, crescimento inicial e alterações anatômicas de Lactuca sativa L. As plantas utilizam vários mecanismos para reduzir os impactos causados pela ação antrópica, como elementos xenobióticos de solos e água contaminada por metais pesados. Esses metais foram fornecidos como nitrato de chumbo e nitrato de prata e foram estabelecidos os seguintes tratamentos: controle para ambos os metais, dose máxima de metais pesados, para solos cultiváveis, permitida pelo Conselho Nacional do Meio Ambiente (Ag = 25mg.Kg-1, Pb = 180mg.Kg-1), dobro (Ag = 50mg.Kg-1, Pb = 360mg.Kg-1) e triplo (Ag = 75mg.Kg-1, Pb = 540 mg.Kg -1) desta dosagem. Foram realizados testes de vigor e germinação das sementes e possíveis alterações anatômicas nas folhas e raízes das plantas de alface. A espécie apresentou alta capacidade de germinar sob estresse de Ag e Pb, e a germinação nunca foi completamente inibida; entretanto, a germinação diminuiu com o aumento das concentrações de Pb, mas não sob estresse de Ag. O uso de doses crescentes dos metais, reduziu o vigor das sementes e aumentou o teor de clorofila. Também foi observado aumento da biomassa nas plantas a partir dos tratamentos submetidos ao Pb. Os efeitos fitotóxicos dos metais foram mais acentuados aos 15 dias após a semeadura. Anatomicamente, L. sativa foi influenciada pelas concentrações de metais, e teve uma redução de até 79,9% na espessura da epiderme radicular na maior concentração de Pb, embora algumas estruturas não tenham sofrido alterações significativas. Os resultados sugerem que L. sativa apresenta tolerância a altas concentrações de metais pesados, mostrando possíveis mecanismos para superar o estresse causado por esses metais. Nesta pesquisa a alface possivelmente utilizou o mecanismo de exclusão de metais retendo Pb e Ag nas raízes preservando o aparato fotossintético na parte aérea das plantas. De forma geral o elemento químico Pb se mostrou mais tóxico que Ag, nestas condições experimentais.

A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals.

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-1, Pb = 180 mg. Kg-1), double (Ag = 50 mg. Kg-1, Pb = 360 mg. Kg-1) and triple (Ag = 75 mg. Kg-1, Pb = 540 mg. Kg -1) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Lead exposure affects cephalic morphogenesis and neural crest cells in Gallus gallus embryo.

The morphogenesis of the head of vertebrates is a process that involves rapid growth and dynamic movements of various cell populations, including the neural crest cells (NCC). These pluripotent cells generated during neurulation have high proliferative and migratory capacity but xenobiotic agents can affect these migratory periods and cause congenital malformations. Lead (Pb) is the most common toxic metal in the environment and a potent teratogen that can affect growth and induce malformations. Despite the known toxic effects of Pb, there is a gap in knowledge about the impact of realistic concentrations of Pb at critical periods of early development. Here, we evaluated mortality, embryonic morphology, NCC migration, and the amount of Pb deposition in chicken embryos after 3 to 4 days of exposure. One of the most interesting observations in this study is that only about 34% of the injected Pb was present in the embryos after 4 days. We observed that exposure to Pb, even under low concentrations, increased mortality and the occurrence of malformations during embryonic development, especially in the cephalic region (CR). Although Pb was found widely distributed in the CR, no relation between its presence and the migration routes of cephalic NCC was observed. But the number of NCC and their migratory distance were reduced. These changes are consistent and explain the morphological anomalies described in this study, which also correlates with the morphofunctional abnormalities reported in the literature. Therefore, this study highlights the concern of exposure to low concentrations of this metal.

Physiological changes in the hemolymph of juvenile shrimp Litopenaeus vannamei to sublethal nitrite and nitrate stress in low-salinity waters.

Juveniles of the shrimp Litopenaeus vannamei (3.3 ±â€¯0.4 g) were exposed separately to nitrite (0.0, 1.1, 2.6, and 5.3 mg/L nitrogen as nitrite [NO2--N]) and nitrate (0, 90, 225 and 400 mg/L nitrogen as nitrate [NO3--N]) concentrations equivalent to 0, 10, 25, and 50% of the LC50-96 h value of NO2--N and NO3--N in low salinity water (3 g/L). Shrimps responded to nitrite and nitrate according to changes in oxyhemocyanin, glucose, lactate and ion levels in the hemolymph after 6, 12, 24, and 48 h of exposure. Oxyhemocyanin levels decreased with increasing nitrite and nitrate levels and were higher at 50% exposure to the contaminants. Compared to the control, glucose and lactate increased significantly at 50% exposure to nitrite and nitrate, particularly at 12 and 24 h. Na+ in the hemolymph changed with nitrite and nitrate, while K+ only changed ˜with nitrite.

Nitrate level safety to Amazon River shrimp juveniles.

The study's objective was to evaluate the nitrate level safety for Macrobrachium amazonicum juvenile in the laboratory, a potential native species for culture in Brazil. The experiment consisted of six treatments with six replicates in a completely randomized block design: 0, 250, 500, 1,000, 1,500, and 2,000 mg L-1. Physical and chemical water quality parameters were recorded every 12 h, while the shrimp mortalities in the 24-h interval. Except for nitrate, all physical and chemical water quality parameters remained within the ideal range rearing to this species. No deaths were observed during the first 6 h of exposure range 0-500 mg L-1 concentrations. At 250 mg L-1 N-NO3-, the mortality (10%) started from 48 h. At 500 mg L-1 N-NO3-, shrimp mortalities occurred after 24 h, reaching 60% after 72 h. In the treatments with 1000 and 1500 mg L-1 N-NO3- concentrations, dead shrimps can be observed after 24 h, with a mortality rate of 78% and 90% of the population in 96 h, respectively. All shrimps exposed at 2000 mg L-1 died in 96 h. The LC50 values obtained decreased with increasing exposure time. Based on LC50 (96 h), the N-NO3- level safety to M. amazonicum is 48.5 mg L-1.

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.

Acute toxicity of nitrate in Litopenaeus vannamei juveniles at low salinity levels / Toxicidade aguda do nitrato em juvenis de Litopenaeus vannamei em baixa salinidade

Different technologies have been developed to improve the performance of Litopenaeus vannamei in low salinity, mainly in super-intensive systems like recirculation and BFT (Biofloc Technology System) systems. However, there is an accumulation of toxic nitrogenous compounds to the shrimps such as nitrate, that at high concentrations and depending on the salinity of the culture water can be lethal. Acute toxicity tests allow to analyze the relationship between the compound and other abiotic or biotic variables. The aim of this research was to determine the acute toxicity and safety level of nitrate (N-NO3 -) for juveniles of L. vannamei at salinities of 5 and 10g.L-1. For salinity of 5g.L-1, a control and 5 treatments were tested, with nitrate concentrations of 100, 500, 1500, 2500 and 3500mg.L-1.For salinity of 10mg.L-1, a 4500mg.L-1nitrate concentration was added. Juveniles were exposed to concentrations during 24, 48, 72, 96 hours in static system. The Mean Lethal Concentration (LCC50) was calculated and the recommended safety level for L. vannamei cultivation is 60.05 and 127.61mg.L-1 of nitrate for salinities 5 and 10g.L-1, respectively.(AU)

Diferentes tecnologias foram desenvolvidas para melhorar o desempenho do Litopenaeus vannamei em baixa salinidade, principalmente em sistemas super intensivos como sistema de recirculação e BFT (Biofloc Technology System). No entanto, há um acúmulo de compostos nitrogenados tóxicos aos camarões, como o nitrato, que em altas concentrações e dependendo da salinidade da água pode ser letal. Os testes de toxicidade aguda permitem analisar a relação entre o composto e outras variáveis ​​abióticas ou bióticas. O objetivo deste trabalho foi determinar a toxicidade aguda e o nível de segurança do nitrato (N-NO3 -) em juvenis de L. vannamei nas salinidades de 5 e 10g.L-1. Para a salinidade de 5g.L-1, um controle e cinco tratamentos foram testados, com concentrações de nitrato 100, 500, 1500, 2500 e 3500mg.L-1. Para salinidade de 10mg.L-1, foi adicionada uma concentração de nitrato de 4500mg.L-1. Os juvenis foram expostos às concentrações durante 24, 48, 72, 96 horas em sistema estático. A Concentração Letal Média (CL50) foi calculada e o nível de segurança recomendado para o cultivo de L. vannamei é de 60,05 e 127,61mg.L-1 de nitrato para salinidadesde 5 e 10g.L-1, respectivamente.(AU)

Subacute intoxication with sodium nitrate induces hematological and biochemical alterations and liver injury in male Wistar rats.

Nitrate pollution has emerged as a problem of great importance because in recent years, the levels of nitrate in soil and groundwater have increased, mainly through anthropogenic activities, such as the use of fertilizers in agriculture, domestic wastewater and septic tanks, industrial waste and deforestation. In animals, nitrate reduction to nitrite (NO2) and nitric oxide (NO) promote the formation of methemoglobin in the blood and the generation of highly reactive intermediates that induce oxidative stress in target organs. Exposition to nitrates has been associated with methemoglobinemia, reproductive toxicity, metabolic and endocrine alterations and cancer. This study analyzed acute intoxication with sodium nitrate (NaNO3) in male Wistar rats, aged 12-16 weeks. Four groups with n = 10 rats each were formed: group 1 was the control, and group 2, group 3 and group 4 were treated for 10 days with intragastric doses of 19, 66 and 150 mg/kg/d NaNO3, respectively. Hematological, metabolic and histological biomarkers in the liver were analyzed. The results showed high percentages of methemoglobin, an increase in NO2 in the plasma and an accumulation in the liver. Moreover, there were high counts of white blood cells and platelets in all treated groups. Additionally, there was an increase in the spleen weight in group 4. High levels of glucose, triglycerides, lactate dehydrogenase (LDH), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were observed and were significantly increased in groups 3 and 4. For oxidative stress biomarkers, there were increases in Thiobarbituric Acid Reactive Substances (TBARS), total GSH and SOD activity, mainly in group 4. Changes in mitochondrial activity were not significant. Histopathological analyses of the liver showed inflammation, infiltration of mononuclear cells, steatosis, ischemia and necrosis, and these findings were more evident at high doses of NaNO3 in which high of S-nitrosylation were found. In conclusion, NaNO3 was reduced to NO2, thereby inducing methemoglobinemia, whereas other reactive species generated oxidative stress, causing hematological and metabolic alterations and injury to the liver.

Nitrate and nitrite poisoning in sheep and goats caused by ingestion of Portulaca oleracea / Intoxicação por nitratos e nitritos em ovinos e caprinos causada pela ingestão de Portulaca oleracea

Three outbreaks of poisoning by Portulaca oleracea were reported in sheep and goats in Northeast Brazil. In the first outbreak, 8 out of 20 sheep were affected and later died. In the second outbreak, three goats and one sheep died out of a flock of 30 animals that included both species. In the third outbreak, two out of 19 sheep were affected, and they recovered after a treatment of 2% methylene blue at a dose of 4 mg/kg body weight. In the first and second outbreaks, the animals ingested P. oleracea after it was cut and offered in feeders. In the third outbreak, the flock was grazing in an area that had been invaded by the plant. To determine the toxicity, P. oleracea was administered experimentally at a dose of 80g/kg of body weight to seven sheep, weighing 19-30 kg. One control sheep received green grass. One to four hours after P. oleracea ingestion, the animals showed clinical signs of poisoning characterized by cyanotic mucous membranes, bloat, ruminal pH of 8-9, pollakiuria, aerophagia, involuntary movements of the upper lip, apathy, tachypnea and tachycardia. Five animals recovered, including one that was treated with 1% methylene blue, and two animals died. During necropsy, the mucous membranes were brownish, and the blood was dark brown. Diphenylamine tests of the plant and of rumen contents were positive for nitrates. Positive results for nitrates were also found in 24 samples of P. oleracea that were collected in different places in the states of Pernambuco and Paraíba. We conclude that P. oleracea accumulates nitrates at toxic levels and may cause poisoning in sheep and goats.(AU)

Relatam-se três surtos de intoxicação por Portulaca oleracea em ovinos e caprinos no Nordeste do Brasil. No primeiro surto morreram oito de 20 ovinos. No segundo morreram três caprinos e um ovino de um total de 30 animais das duas espécies. No terceiro surto foram afetadas duas ovelhas de um rebanho de 19 animais, que se recuperaram após o tratamento com azul de metileno a 2% na dose de 4 mg/kg/vivo. Nos surtos 1 e 2 os animais ingeriram P. oleracea cortada e oferecida no coxo e no surto 3 estavam pastoreando em uma área invadida pela planta. Para determinar a toxicidade de P. oleracea foram utilizados oitos ovinos, sendo sete experimentais e um controle, com peso entre 19 e 30 kg. A planta foi administrada por via oral, na dose de 80 g/kg/peso corporal. O animal controle recebeu capim verde e concentrado. Entre uma a quatro horas após a ingestão da planta os animais apresentaram sinais clínicos caracterizados por mucosas cianóticas, timpanismo gasoso, pH ruminal de 8-9, polaquiúria, aerofagia, movimentos involuntários do lábio superior, apatia, taquipnéia e taquicardia. Cinco animais se recuperaram, incluindo um que foi tratado com azul de metileno a 2%, e dois morreram. Na necropsia observaram-se mucosas de coloração marrom e sangue marrom escuro. O teste de difenilamina realizado na planta e no conteúdo ruminal foi positivo para nitratos. Resultados positivos para nitratos foram detectados em 24 amostras coletadas em diferentes locais dos estados de Pernambuco e Paraíba. Conclui-se que P. oleracea acumula nitratos em níveis tóxicos, e quando ingerida por ovinos e caprinos pode provocar intoxicação e morte.(AU)

Nitrate and nitrite poisoning in sheep and goats caused by ingestion of Portulaca oleracea / Intoxicação por nitratos e nitritos em ovinos e caprinos causada pela ingestão de Portulaca oleracea

Three outbreaks of poisoning by Portulaca oleracea were reported in sheep and goats in Northeast Brazil. In the first outbreak, 8 out of 20 sheep were affected and later died. In the second outbreak, three goats and one sheep died out of a flock of 30 animals that included both species. In the third outbreak, two out of 19 sheep were affected, and they recovered after a treatment of 2% methylene blue at a dose of 4 mg/kg body weight. In the first and second outbreaks, the animals ingested P. oleracea after it was cut and offered in feeders. In the third outbreak, the flock was grazing in an area that had been invaded by the plant. To determine the toxicity, P. oleracea was administered experimentally at a dose of 80g/kg of body weight to seven sheep, weighing 19-30 kg. One control sheep received green grass. One to four hours after P. oleracea ingestion, the animals showed clinical signs of poisoning characterized by cyanotic mucous membranes, bloat, ruminal pH of 8-9, pollakiuria, aerophagia, involuntary movements of the upper lip, apathy, tachypnea and tachycardia. Five animals recovered, including one that was treated with 1% methylene blue, and two animals died. During necropsy, the mucous membranes were brownish, and the blood was dark brown. Diphenylamine tests of the plant and of rumen contents were positive for nitrates. Positive results for nitrates were also found in 24 samples of P. oleracea that were collected in different places in the states of Pernambuco and Paraíba. We conclude that P. oleracea accumulates nitrates at toxic levels and may cause poisoning in sheep and goats.(AU)

Relatam-se três surtos de intoxicação por Portulaca oleracea em ovinos e caprinos no Nordeste do Brasil. No primeiro surto morreram oito de 20 ovinos. No segundo morreram três caprinos e um ovino de um total de 30 animais das duas espécies. No terceiro surto foram afetadas duas ovelhas de um rebanho de 19 animais, que se recuperaram após o tratamento com azul de metileno a 2% na dose de 4 mg/kg/vivo. Nos surtos 1 e 2 os animais ingeriram P. oleracea cortada e oferecida no coxo e no surto 3 estavam pastoreando em uma área invadida pela planta. Para determinar a toxicidade de P. oleracea foram utilizados oitos ovinos, sendo sete experimentais e um controle, com peso entre 19 e 30 kg. A planta foi administrada por via oral, na dose de 80 g/kg/peso corporal. O animal controle recebeu capim verde e concentrado. Entre uma a quatro horas após a ingestão da planta os animais apresentaram sinais clínicos caracterizados por mucosas cianóticas, timpanismo gasoso, pH ruminal de 8-9, polaquiúria, aerofagia, movimentos involuntários do lábio superior, apatia, taquipnéia e taquicardia. Cinco animais se recuperaram, incluindo um que foi tratado com azul de metileno a 2%, e dois morreram. Na necropsia observaram-se mucosas de coloração marrom e sangue marrom escuro. O teste de difenilamina realizado na planta e no conteúdo ruminal foi positivo para nitratos. Resultados positivos para nitratos foram detectados em 24 amostras coletadas em diferentes locais dos estados de Pernambuco e Paraíba. Conclui-se que P. oleracea acumula nitratos em níveis tóxicos, e quando ingerida por ovinos e caprinos pode provocar intoxicação e morte.(AU)

[Exposure to nitrates in drinking water and its association with thyroid gland dysfunction]. / Exposición a nitratos en agua y su relación con disfunción de la glándula tiroides: revisión sistemática ¿Existen riesgos para la salud de la población?

BACKGROUND: Nitrate exposure may be associated with thyroid gland dysfunction. AIM: To review the available evidence about the relationship between nitrates in drinking water and thyroid gland dysfunction. MATERIAL AND METHODS: A wide search was performed using Medline, Cochrane, Lilacs, IBECS and Scielo databases using pertinent keywords, finding a total of 66 related studies. After filtering and in depth reviewing, a total of 12 studies were included in this review. RESULTS: The main results reveal the importance of this ion for human health, finding evidence both in animals and human beings that suggest pathological changes in the gland as its relationship with the occurrence of subclinical hypothyroidism, and potentially cancer of the thyroid gland. In Chile, nitrate is not considered a critical contaminant so its regular measuring and control is not enforced. CONCLUSIONS: In light of the present review we believe that there is evidence to consider nitrate as a critical contaminant whose measurement, registration and correct implementation of valid policies would have a direct benefit for the population of this country. Without this information, it is not possible to quantify the damage to human health, especially in vulnerable groups residing in areas at greatest risk of exposure.

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.

Exposición a nitratos en agua y su relación con disfunción de la glándula tiroides: revisión sistemática ¿Existen riesgos para la salud de la población? / Exposure to nitrates in drinking water and its association with thyroid gland dysfunction

Background: Nitrate exposure may be associated with thyroid gland dysfunction. Aim: To review the available evidence about the relationship between nitrates in drinking water and thyroid gland dysfunction. Material and Methods: A wide search was performed using Medline, Cochrane, Lilacs, IBECS and Scielo databases using pertinent keywords, finding a total of 66 related studies. After filtering and in depth reviewing, a total of 12 studies were included in this review. Results: The main results reveal the importance of this ion for human health, finding evidence both in animals and human beings that suggest pathological changes in the gland as its relationship with the occurrence of subclinical hypothyroidism, and potentially cancer of the thyroid gland. In Chile, nitrate is not considered a critical contaminant so its regular measuring and control is not enforced. Conclusions: In light of the present review we believe that there is evidence to consider nitrate as a critical contaminant whose measurement, registration and correct implementation of valid policies would have a direct benefit for the population of this country. Without this information, it is not possible to quantify the damage to human health, especially in vulnerable groups residing in areas at greatest risk of exposure.

Identification of up-regulated genes from the metal-hyperaccumulator aquatic fern Salvinia minima Baker, in response to lead exposure.

Lead (Pb) is one of the most serious environmental pollutants. The aquatic fern Salvinia minima Baker is capable to hyper-accumulate Pb in their tissues. However, the molecular mechanisms involved in its Pb accumulation and tolerance capacity are not fully understood. In order to investigate the molecular mechanisms that are activated by S. minima in response to Pb, we constructed a suppression subtractive hybridization library (SSH) in response to an exposure to 40µM of Pb(NO3)2 for 12h. 365 lead-related differentially expressed sequences tags (ESTs) were isolated and sequenced. Among these ESTs, 143 unique cDNA (97 were registered at the GenBank and 46 ESTs were not registered, because they did not meet the GenBank conditions). Those ESTs were identified and classified into 3 groups according to Blast2GO. In terms of metabolic pathways, they were grouped into 29 KEGG pathways. Among the ESTs, we identified some that might be part of the mechanism that this fern may have to deal with this metal, including abiotic-stress-related transcription factors, some that might be involved in tolerance mechanisms such as ROS scavenging, membrane protection, and those of cell homeostasis recovery. To validate the SSH library, 4 genes were randomly selected from the library and analyzed by qRT-PCR. These 4 genes were transcriptionally up-regulated in response to lead in at least one of the two tested tissues (roots and leaves). The present library is one of the few genomics approaches to study the response to metal stress in an aquatic fern, representing novel molecular information and tools to understand the molecular physiology of its Pb tolerance and hyperaccumulation capacity. Further research is required to elucidate the functions of the lead-induced genes that remain classified as unknown, to perhaps reveal novel molecular mechanisms of Pb tolerance and accumulation capacity in aquatic plants.

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.

Chronic effects of nitrogenous compounds on survival and growth of juvenile pink shrimp.

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.

Ecotoxicological risks of calcium nitrate exposure to freshwater tropical organisms: Laboratory and field experiments.

This study aimed to analyze laboratory and field data to assess the ecotoxicological risks of calcium nitrate exposure to freshwater tropical biota. Short-term laboratorial tests resulted in estimated EC50 values of 76.72 (67.32-86.12)mg N-NO3₋ L(-1) for C. silvestrii and 296.46 (277.16-315.76) mg N-NO3₋ L(-1) for C. xanthus. Long-term laboratorial tests generated IC25 values of 5.05 (4.35-5.75) and 28.73 (26.30-31.15) mg N-NO3₋ L(-1) for C. silvestrii and C. xanthus, respectively. The results from in situ mesocosm experiments performed in the Ibirité reservoir (a tropical eutrophic urban water body located in SE Brazil) indicated that C. silvestrii and C. xanthus were not under severe deleterious acute impact due to the treatment because the higher nitrate concentrations determined were 5.2 mg N-NO3₋ L(-1) (t=24 h; sediment-water interface) and 17.5 mg N-NO3₋ L(-1) (t=600 h; interstitial water). However, an abrupt decrease in the densities of Cyanophyceae members and other benthic taxa was observed. In summary, the present work contributes greatly to the toxicity data linked to two taxonomically distinct organisms that have never been screened for calcium nitrate sensitivity. Furthermore, considering the problem of the management and restoration of eutrophic environments, our study reports a comprehensive field assessment that allows the elucidation of the possible toxic impacts caused by the addition of calcium nitrate (a remediation technique) on aquatic and benthic organisms as well as the implications on the aquatic ecosystem as a whole, which may greatly allow expanding the current knowledgebase on the topic.

The chronic toxicity of ammonia, nitrite and nitrate on juvenile Farfantepenaeus brasiliensis (Crustacea: Decapoda) / Toxicidade crônica da amônia, nitrito e nitrato em juvenis de Farfantepenaeus brasiliensis (Crustacea: Decapoda)

In general, the adverse effect of a chemical compound present in water varies with the concentration and time of exposure to the compound, the nature of the chemical species and age of the exposed organisms. Thus, nitrogen does not necessarily cause adverse effects on shrimp, but may, instead, promote sub-lethal effects by long-term exposure. Juvenile Farfantepenaeus brasiliensis (initial mean weight = 0.61 g ± 0.07) were exposed to sub-lethal concentrations of ammonia (0.44 and 0.88 mg L-1), nitrite (5.30 and 10.60 mg L-1) and nitrate (45.60 and 91.20 mg L-1) corresponding to the safe levels for the species. After 40 days of exposure of juveniles to ammonia, nitrite and nitrate, all groups differed significantly (p 0,05) from the control group regarding the growth and survival. Based on the results, it was determined that the shrimp F. brasiliensis was susceptible to nitrogen compounds in concentrations equivalent to supposedly safe levels previously proposed for the specie. Thus, the security levels of ammonia, nitrite and nitrate for pink shrimp juveniles were 0.88 mg L-1, 10.60 mg L-1 and 91.20 mg L-1, respectively.

Em geral, o efeito adverso de um composto químico presente na água varia com a concentração, o tempo de exposição ao composto, à natureza do produto químico e a idade das espécies de organismos expostos. Assim, o nitrogênio não necessariamente causa efeitos adversos, mas pode, em vez disso, promover efeitos subletais por meio da exposição em longo prazo. Juvenis de Farfantepenaeus brasiliensis (peso médio inicial = 0,61 g ± 0,07) foram expostos a concentrações subletais de amônia (0,44 e 0,88 mg L-1), nitrito (5,30 e 10,60 mg L-1) e nitrato (45,60 e 91,20 mg L-1) correspondente aos "níveis de segurança" para a espécie. Após 40 dias de exposição dos juvenis à amônia, nitrito e nitrato, todos os grupos diferiram significativamente (p 0,05) do grupo controle em relação ao crescimento e sobrevivência. Com base nos resultados, o camarão F. brasiliensis foi susceptível aos compostos nitrogenados em concentrações equivalentes aos níveis supostamente seguros anteriormente propostos para a espécie. Assim, os níveis de segurança de amônia, nitrito e nitrato propostos para juvenis de camarão-rosa são 0,88 mg L-1, 10,60 mg L-1 e 91,20 mg L-1, respectivamente.

The chronic toxicity of ammonia, nitrite and nitrate on juvenile Farfantepenaeus brasiliensis (Crustacea: Decapoda) / Toxicidade crônica da amônia, nitrito e nitrato em juvenis de Farfantepenaeus brasiliensis (Crustacea: Decapoda)

In general, the adverse effect of a chemical compound present in water varies with the concentration and time of exposure to the compound, the nature of the chemical species and age of the exposed organisms. Thus, nitrogen does not necessarily cause adverse effects on shrimp, but may, instead, promote sub-lethal effects by long-term exposure. Juvenile Farfantepenaeus brasiliensis (initial mean weight = 0.61 g ± 0.07) were exposed to sub-lethal concentrations of ammonia (0.44 and 0.88 mg L-1), nitrite (5.30 and 10.60 mg L-1) and nitrate (45.60 and 91.20 mg L-1) corresponding to the safe levels for the species. After 40 days of exposure of juveniles to ammonia, nitrite and nitrate, all groups differed significantly (p 0,05) from the control group regarding the growth and survival. Based on the results, it was determined that the shrimp F. brasiliensis was susceptible to nitrogen compounds in concentrations equivalent to supposedly safe levels previously proposed for the specie. Thus, the security levels of ammonia, nitrite and nitrate for pink shrimp juveniles were 0.88 mg L-1, 10.60 mg L-1 and 91.20 mg L-1, respectively.(AU)

Em geral, o efeito adverso de um composto químico presente na água varia com a concentração, o tempo de exposição ao composto, à natureza do produto químico e a idade das espécies de organismos expostos. Assim, o nitrogênio não necessariamente causa efeitos adversos, mas pode, em vez disso, promover efeitos subletais por meio da exposição em longo prazo. Juvenis de Farfantepenaeus brasiliensis (peso médio inicial = 0,61 g ± 0,07) foram expostos a concentrações subletais de amônia (0,44 e 0,88 mg L-1), nitrito (5,30 e 10,60 mg L-1) e nitrato (45,60 e 91,20 mg L-1) correspondente aos "níveis de segurança" para a espécie. Após 40 dias de exposição dos juvenis à amônia, nitrito e nitrato, todos os grupos diferiram significativamente (p 0,05) do grupo controle em relação ao crescimento e sobrevivência. Com base nos resultados, o camarão F. brasiliensis foi susceptível aos compostos nitrogenados em concentrações equivalentes aos níveis supostamente seguros anteriormente propostos para a espécie. Assim, os níveis de segurança de amônia, nitrito e nitrato propostos para juvenis de camarão-rosa são 0,88 mg L-1, 10,60 mg L-1 e 91,20 mg L-1, respectivamente.(AU)

Potential application in mercury bioremediation of a marine sponge-isolated Bacillus cereus strain Pj1.

Sponges are sessile marine invertebrates that can live for many years in the same location, and therefore, they have the capability to accumulate anthropogenic pollutants such as metals over a long period. Almost all marine sponges harbor a large number of microorganisms within their tissues. The Bacillus cereus strain Pj1 was isolated from a marine sponge, Polymastia janeirensis, and was found to be resistant to 100 µM HgCl(2) and to 10 µM methylmercury (MeHg). Pj1 was also highly resistant to other metals, including CdCl(2) and Pb(NO(3))(2), alone or in combination. The mer operon was located on the bacterial chromosome, and the volatilization test indicated that the B. cereus Pj1 was able to reduce Hg(2+)-Hg(0). Cold vapor atomic absorption spectrometry demonstrated that Pj1 volatilized 80 % of the total MeHg that it was exposed to and produced elemental Hg when incubated with 1.5 µM MeHg. Pj1 also demonstrated sensitivity to all antibiotics tested. In addition, Pj1 demonstrated a potential for biosurfactant production, presenting an emulsification activity better than synthetic surfactants. The results of this study indicate that B. cereus Pj1 is a strain that can potentially be applied in the bioremediation of HgCl(2) and MeHg contamination in aquatic environments.