Commentary of the SKLM to the EFSA opinion on risk assessment of N-nitrosamines in food.

Dietary exposure to N-nitrosamines has recently been assessed by the European Food Safety Authority (EFSA) to result in margins of exposure that are conceived to indicate concern with respect to human health risk. However, evidence from more than half a century of international research shows that N-nitroso compounds (NOC) can also be formed endogenously. In this commentary of the Senate Commission on Food Safety (SKLM) of the German Research Foundation (DFG), the complex metabolic and physiological biokinetics network of nitrate, nitrite and reactive nitrogen species is discussed with emphasis on its influence on endogenous NOC formation. Pioneering approaches to monitor endogenous NOC have been based on steady-state levels of N-nitrosodimethylamine (NDMA) in human blood and on DNA adduct levels in blood cells. Further NOC have not been considered yet to a comparable extent, although their generation from endogenous or exogenous precursors is to be expected. The evidence available to date indicates that endogenous NDMA exposure could exceed dietary exposure by about 2-3 orders of magnitude. These findings require consolidation by refined toxicokinetics and DNA adduct monitoring data to achieve a credible and comprehensive human health risk assessment.

Ecotoxicity characterization assisted performance assessment of electro-bioremediation reactors for nitrate and arsenite elimination.

The performance of combined reduction of nitrate (NO3 - ) to dinitrogen gas (N2 ) and oxidation of arsenite (As[III]) to arsenate (As[V]) by a bioelectrochemical system was assessed, supported by ecotoxicity characterization. For the comprehensive toxicity characterization of the untreated model groundwater and the treated reactor effluents, a problem-specific ecotoxicity test battery was established. The performance of the applied technology in terms of toxicity and target pollutant elimination was compared and analyzed. The highest toxicity attenuation was achieved under continuous flow mode with hydraulic retention time (HRT) = 7.5 h, with 95%, nitrate removal rate and complete oxidation of arsenite to arsenate. Daphnia magna proved to be the most sensitive test organism. The results of the D. magna lethality test supported the choice of the ideal operational conditions based on chemical data analysis. The outcomes of the study demonstrated that the applied technology was able to improve the groundwater quality in terms of both chemical and ecotoxicological characteristics. The importance of ecotoxicity evaluation was also highlighted, given that significant target contaminant elimination did not necessarily lower the environmental impact of the initial, untreated medium, in addition, anomalies might occur during the technology operational process which in some instances, could result in elevated toxicity levels.

Assessment of the effects of sodium nitrate (NaNO3) on the reproductive system, liver, pancreas, and kidney of male rats.

Nitrate (NO3) toxicity is a serious global issue that results in impairment of physiological systems of our body. The present study aimed to investigate the effects of different concentration of NaNO3 (10, 100, 500 and 1000 mg/kg bw) on the male reproductive system, liver, kidney and pancreas. Adult male Wistar rats were divided into five groups of five animals each (n = 5). The first group served as controls. The second, third, fourth and fifth groups of rat were orally intubated with 10, 100, 500 and 1000 mg/kg bw of NaNO3 for 52 days. After the treatment period, the rats were sacrificed and NO3 induced alterations on selected organs were assessed. There was a dose dependent decrease in sperm motility, serum concentration of testosterone, body weight and organ weight, and increase in abnormal sperm morphology in the NaNO3 treated groups compared with the controls. Further, histological analysis confirmed that NO3 induced toxicity. Shrunken seminiferous tubules and loss of spermatids in testes, shrinkage of acinar cells of the pancreas, sinusoidal congestion and necrosis in the liver, atrophy of glomeruli and congestion of renal tubules of the kidney were the histological alterations observed in rats treated with100 and 500 mg/kg NaNO3. However, 100% mortality was observed in rats treated with 1000 mg/kg NaNO3. The present study clearly demonstrated the toxic effects of NaNO3 on both the reproductive system and other organs of the body. The study might inform human studies; where in the chances of male infertility may be more a problem for individuals in areas with NO3-rich ground water.

Aggregating exposures and toxicity equivalence approach into an integrated probabilistic dietary risk assessment for perchlorate, nitrate, and thiocyanate: Results from the National food monitoring study and National Food Consumption Database.

Perchlorate, nitrate, and thiocyanate, namely thyroid disrupting chemicals (TDCs), are found ubiquitously in the environment, leading to broad human exposure and primary uptake through the food web and drinking water. TDCs are all competitive inhibitors of thyroid iodide uptake activity, but limited studies have assessed the cumulative risk of dietary exposure to multiple TDCs. Thus, in this study, we analyzed the individual exposure risk from 310 food samples in 11 categories, and also assessed the cumulative health risks from TDCs for the Taiwanese population using a perchlorate equivalent concentration (PEC) approach. Consequently, this study not only demonstrated the non-carcinogenic health risks from individual exposure but also highlighted that the cumulative exposure to these TDCs may adversely affect human thyroid functioning. Vegetables, livestock, fruits, and dairy products are the most susceptible to PEC exposure. We highlighted nitrate as the main contributor to PEC exposure. Finally, controlling the overall TDC concentrations from vegetables, livestock, fruits, and dairy products is emphasized in this study. This is the first study to conduct a cumulative risk assessment of dietary exposure to TDCs using the PEC approach for the Taiwanese population through probabilistic and sensitivity analyses.

Groundwater quality evaluation and human health risks assessment using the WQI, NPI and HQnitrate models: case of the Sfax intermediate aquifer, Sahel Tunisia.

Groundwater is a vital natural resource required to satisfy the domestic and agricultural needs. In general, human health is linked to the quality of the consumed water. For instance, long-term exposure to high nitrate levels in groundwater may cause problems. Hence, the present study was conducted to assess the nitrate contamination of groundwater as well as its related health risks for the inhabitants of the Sfax region, Sahel Tunisia. Irrigation groundwater suitability has been evaluated with sodium content (%Na), electrical conductivity (EC), magnesium hazard (MH), sodium adsorption ratio (SAR), permeability index (PI), Kelly's ratio (KR) and soluble sodium percent (SSP). The results indicate that the selected groundwater is characterized by low to moderate quality for irrigation. Furthermore, the drinking water quality index (DWQI) was assessed using potential of hydrogen (pH), total dissolved solids (TDS), magnesium (Mg2+), calcium (Ca2+), sodium (Na+), chloride (Cl-), sulfate (SO42-), potassium (K+), bicarbonate (HCO3-) and nitrate (NO3-). The results indicate that 3.63% of samples have good quality of water, while 41.82% have poor to very poor water quality and the rest (54.55%) are unfit for drinking. The nitrate pollution index (NPI) model revealed that about 42% of the samples present significant to very significant type of pollution. Based on human health risk assessment, the children are at higher risks compared to the other affected groups. The obtained results could be used as a basic document for realistic management of groundwater quality and to provide an overview for decision-making authorities to take necessary actions.

Nitrate contamination in drinking water and colorectal cancer: Exposure assessment and estimated health burden in New Zealand.

BACKGROUND: Epidemiological evidence in multiple jurisdictions has shown an association between nitrate exposure in drinking water and an increased risk of colorectal cancer (CRC). OBJECTIVE: We aimed to review the extent of nitrate contamination in New Zealand drinking water and estimate the health and financial burden of nitrate-attributable CRC. METHODS: We collated data on nitrate concentrations in drinking water for an estimated 85% of the New Zealand population (∼4 million people) who were on registered supplies. We estimated nitrate levels for the remaining population (∼600,000 people) based on samples from 371 unregistered (private) supplies. We used the effective rate ratio from previous epidemiological studies to estimate CRC cases and deaths attributable to nitrate in drinking water. RESULTS: Three-quarters of New Zealanders are on water supplies with less than 1 mg/L NO3-N. The population weighted average for nitrate exposure for people on registered supplies was 0.49 mg/L NO3-N with 1.91% (95%CI 0.49, 3.30) of CRC cases attributable to nitrates. This correlates to 49.7 cases per year (95%CI 14.9, 101.5) at a cost of 21.3 million USD (95% 6.4, 43.5 million USD). When combining registered and unregistered supplies, we estimated 3.26% (95%CI 0.84, 5.57) of CRC cases were attributable to nitrates, resulting in 100 cases (95%CI 25.7, 171.3) and 41 deaths (95%CI 10.5, 69.7) at a cost of 43.2 million USD (95%CI 10.9, 73.4). CONCLUSION: A substantial minority of New Zealanders are exposed to high or unknown levels of nitrates in their drinking water. Given the international epidemiological studies showing an association between cancer and nitrate ingestion from drinking water, this exposure may cause an important burden of preventable CRC cases, deaths, and economic costs. We consider there is sufficient evidence to justify a review of drinking water standards. Protecting public health adds to the strong environmental arguments to improve water management in New Zealand.

Probabilistic risk and benefit assessment of nitrates and nitrites by integrating total diet study-based exogenous dietary exposure with endogenous nitrite formation using toxicokinetic modeling.

The impacts of dietary nitrates and nitrites on human health have been a controversial topic for many years. However, the risk and benefit assessment of nitrates and nitrites is complicated by the large variation in nitrate and nitrite intake among people and the endogenous nitrite formation in the body. This study conducted a probabilistic risk-benefit assessment of dietary nitrates and nitrites based on internal dose by integrating exogenous and endogenous exposures with human trial data on cardiovascular benefits. A total diet study was carried out to quantify the age-specific dietary intakes of nitrates and nitrites. A previously well-validated human toxicokinetic model was used to predict internal doses for different age groups. In addition, the integrated approach was applied to different populations from different countries/regions based on reported exposure estimates to conduct a comprehensive risk-benefit assessment of dietary nitrates and nitrites. The results demonstrated that vegetable consumption was the main contributor to the internal nitrate and nitrite levels in all age groups. Exposure to nitrates and nitrites exceeding acceptable daily intakes in a variety of foods showed cardiovascular benefits. The probabilistic risk assessment showed that the exposure to nitrates and nitrites did not pose an appreciable health and safety risk. Therefore, the present results suggest that dietary nitrates and nitrites have clear cardiovascular benefits that may outweigh potential risks. Our analysis contributes significantly to addressing the controversy regarding risks and benefits from dietary nitrates and nitrites, and our approach could be applied to other dietary constituents with the potential for both risks and benefits.

Health Risk Assessment for Exposure to Nitrate in Drinking Water in Central Java, Indonesia.

Since 2005, over 30 epidemiological studies have evaluated the association between nitrate in drinking water and adverse health outcomes. Conditions that lead to nitrate pollution in water, such as open defecation, the proximity of septic tanks to water sources, and the use of inorganic fertilizer, are rampant in Indonesia, which has experienced little research evaluating nitrate in drinking water. We conducted a health risk assessment for exposure to nitrate in drinking water and evaluated the nitrate concentration in key water sources in two villages of rural Central Java, Indonesia. The nitrate concentrations in the drinking water ranged from 3.55 mg/L to 26.75 mg/L as NO3-. Daily nitrate intake estimates, calculated at 50% and 95% exposure to the maximum nitrate concentration of the drinking water in both villages, were above the levels associated with birth defects, colorectal cancer, and thyroid conditions observed in other studies. There was a large variation in nitrate concentrations between and within the villages at different water sources. Further research into whether these health outcomes exist in rural Central Java, Indonesia will be required to better understand this risk.

Cadmium, chromium, nickel and nitrate accumulation in wheat (Triticum aestivum L.) using wastewater irrigation and health risks assessment.

The wastewater utilization for irrigation purposes is common practice in peri-urban areas located in vicinity of developed cities. This water contains elements like chromium (Cr), nickel (Ni), cadmium (Cd) and nitrate (NO3-N) that poses health risk when exposed to human. In this study effect of wastewater irrigation from Chakara wastewater plant, Faisalabad on growth of wheat and health risks was assessed. Pot experiment was conducted at Institute of Soil and Environmental, University of Agriculture, Faisalabad using different concentration of wastewater as treatment 100% tap water, 25% wastewater + 75% tap water, 50% wastewater + 50% tap water, 75% wastewater + 25% tap water, 100% wastewater. The results indicated that the wastewater irrigation negatively effects the plant growth and physiological parameters. The minimum plant height, grain weight, spike length, osmotic potential and SPAD values were recorded 50.33 cm, 1.47 g plant-1, 7.00 cm, 423 and 38.91 respectively in 100% wastewater irrigation. The risk quotient (RQ TEs) for each toxic element and cumulative risk index (RI TEs) values were calculated. The cadmium risk quotient (Cd RQ) for adults was on margin and value was >1 for in 75% wastewater + 25% tap water and 100% wastewater irrigation, while the RQ for Ni and Cr was <1. Maximum RI TEs values calculated in 100% wastewater irrigation 0.424 and 0.294 for children and adults respectively. Hence it was concluded that wastewater irrigation significantly increased the accumulation rate of metals and nitrate in wheat and cause potential health risks for children and adults.

Non-carcinogenic health risk assessment with source identification of nitrate and fluoride polluted groundwater of Wardha sub-basin, central India.

In order to understand the pollution status of groundwater with geochemical evolution and appraisal of its probable public health risk due to nitrate (NO3-) and fluoride (F-), a total of 93 groundwater samples were collected during pre-monsoon (May) period from Wardha sub-basin, central India. By employing Piper plot, transition from Ca-HCO3 type water (recharge waters) to Na-Cl (saline water) type water through mixed Ca-Na-HCO3, mixed Ca-Mg-Cl (reverse ion exchange waters) and Ca-Cl types (leachate waters), were observed. The Geogenic processes such as silicate, dolomite, halite and carbonate weathering along with calcite precipitation and ion exchange process were identified as major controlling factors for evolution and alteration of groundwater chemistry. The Saturation index highlighted that the groundwater in the area is oversaturated with respect to the mineral calcite and dolomite, and under saturated with gypsum, fluorite and halite. The high NO3- and F- concentration overpassing the permissible limit were found in 54.8% and 18.5% of samples. The plot of F- with Na+/Ca2+, Na+/Mg2+ and F-/Cl- established fluoride bearing rock weathering is responsible for F- contamination. Based on the cluster analysis, the groundwater was grouped into Cluster-I Ca-Na-HCO3 type (61.3%) and Cluster-II Na-Ca-HCO3-Cl type (30.1%). The total hazard index (HI) based on human health risk assessment (HHRA) model for cumulative NO3- and F- toxicity through oral and dermal pathways were computed as 100%, 97.85% and 96.77% for children, female and male populations respectively. The HQ(nitrate) > 1 through ingestion pathway were in 84.95%, 68.82% and 62.37%, and HQ(fluoride) > 1 in 83.87%, 62.37% and 43.01% of the groundwater samples were recorded for children, female and male population respectively. The risk assessment study highlighted very high toxicity and severe health impact of ingestion of contaminated groundwater on public health.

Groundwater nitrate pollution risk assessment of the groundwater source field based on the integrated numerical simulations in the unsaturated zone and saturated aquifer.

Groundwater pollution risk assessment in the groundwater source field (GSF) is crucial to ensure groundwater quality safety. A systematic method of assessing groundwater pollution in the GSF was established by combining the numerical models of groundwater flow and solute transport in the vadose zone and aquifer. It is featured by revealing the paramount fate of contaminant from the surface to receptor "well (wells)" via the pathway of vadose zone and aquifers. The method was verified in the phreatic and semi-confined aquifers of a vital GSF, Beijing-Tianjin-Hebei region (BTHR) in China. Nitrate was selected as the model pollutant. The results indicated that the groundwater pollution risk of the phreatic aquifer was dominated by the mediate level (45.27%), and that the second semi-confined aquifer was mainly ranked as relatively low (30.29%) and mediate (38.17%) levels. The groundwater pollution risk maps of the two aquifers were similar. The high and relatively high risk areas were affected by the high intensities of groundwater pollution sources (GPSIs) or short distances from the pollution sources to the pumping well. The low and relatively low risk areas were controlled by low GPSIs and adequate attenuation and denitrification of nitrate in the aquifer. The groundwater pollution risk in the semi-confined aquifer was lower than that in the phreatic aquifer. The groundwater pollution risk mapping provides a valuable scientific reference for the groundwater pollution prevention and control with the focus on the "pollution source" and "groundwater source field". The proposed method can be further applied to the protections of the GSFs in the BTHR.

Incorporating Exogenous and Endogenous Exposures into Dietary Risk Assessment of Nitrates and Nitrites in Vegetables: A Probabilistic Integrated Toxicokinetic Modeling Approach.

This study aimed to estimate the dietary risk of nitrates and nitrites in vegetables based on internal dose in a probabilistic manner by integrating exogenous exposure based on measured concentrations in vegetables with endogenous exposure using a toxicokinetic (TK) model. We optimized and validated a previous TK model and incorporated Monte Carlo simulations to account for variability across different age populations for predicting internal dose. High levels of nitrates were detected in leafy vegetables (from 545 ± 274 to 1641 ± 873 mg/kg). Nitrite contents of vegetables were generally low (from 1.26 ± 1.40 to 8.20 ± 14.1 mg/kg). The dietary risk was found to be different based on internal versus external dose, suggesting that it is critical to include endogenous nitrite formation into risk assessment. Nitrate and nitrite exposure from vegetables is unlikely to result in appreciable risks for most populations but may be a potential risk for preschoolers.

Groundwater quality evaluation using water quality index (WQI) for drinking purposes and human health risk (HHR) assessment in an agricultural region of Nanganur, south India.

An effort has been made to understand the groundwater quality of Nanganur region for drinking purpose utilizing water quality index (WQI). Nitrate contamination in groundwater was assessed and the associated health risks to rural populations were estimated for different age groups, infants, children and adults in the agricultural region of Nanganur, South India, where residents rely on only groundwater for drinking use. Groundwater was slightly alkaline and moderately hard. The concentration of nitrate in the groundwater ranged from 25 to 198 mg/L, with a mean of 66.14 mg/L, and 61% of groundwater samples exceeded the World Health Organization (WHO) safe limit of 50 mg/L. The WQI values ranged from 92 to 295, with an average of 153, and about 86% of groundwater samples were poor quality for drinking uses. Results showed that the non-carcinogenic health risk for adults ranged from 6.0E-01 to 4.8E+00, for children 8.1E-01 to 6.4E+00, and for infants 9.4E-01 to 7.4E+00, respectively. Health risk caused by excessive intake of nitrate contaminated groundwater for children and infants were 1.15 and 1.75 times larger than that for adults, which suggested that health risk degree of infants have greater health risk than children and adults in the study region. The order of nitrate contribution to non-carcinogenic health risk among the studied age groups was infants > children > adults. Therefore, health risk reduction measures should be implemented to reduce exposure to nitrate contaminated drinking water in the study region.

Assessment, mapping, and management of health risk from nitrate accumulation in onion for Iranian population.

Excess accumulation of nitrate in vegetable crops is normally related to the excessive use of nitrogen fertilizers applied in agricultural fields. This study was performed to evaluate the level of nitrate concentration in onion production in Isfahan province of Iran and the potential health risks associated with high level of nitrate in this crop, as an example of a case-study. The risk was estimated by using the United States Environmental Protection Agency (USEPA) method and presented as risk maps. Additionally, the impact of an improved fertilizer management practice in selected vulnerable fields for reducing nitrate concentration in onion production was evaluated. For this assessment, four different onion fields in which onion bulb nitrate concentration was higher than the safety level (70 mg kg-1 FW), provided by Welch (2003) in a survey study, were chosen. The risk maps showed that despite higher risk possibility in adults, in comparison with children, all values were lower than the acceptable level. Further, the split application of nitrogen fertilizers in all fields significantly reduced plant nitrate concentration with no reduction in yield and even observed increase in the overall economic yield production. Therefore, considering crop yields and nitrate accumulation impacts, improved nitrogen management could provide an opportunity to promote onion crop production while reducing negative health effects in high-risk regions in Isfahan province of Iran.

In situ assessment of Karaj River genotoxic impact with the alkaline comet assay and micronucleus test, on feral brown trout (Salmo trutta fario).

The in situ evaluation of the genotoxic impact of the Karaj River was performed using the comet and micronucleus (MN) assays in erythrocytes, liver, gill and kidney of indigenous brown trout, Salmo trutta fario from three different stations, including Varangerud, Asara and Purkan. The results showed that DNA damage significantly increased in sampled fish erythrocytes, liver and gill from low levels in the upstream river (Varangerud) via intermediate levels in downstream (Purkan) to high levels in the middle of the river (Asara), correlating with the river increasing pollution gradient. Gill was the most sensitive tissue followed by blood and liver. Kidney did not respond to the genotoxic gradient of the river. MN test (as a complementary assay) of liver cells of fish was a sensitive biomarker of genotoxic exposure. MN test in blood, gill and kidney did not reflect the genotoxic condition of the river.

Risk assessment and monitoring programme of nitrates through vegetables in the Region of Valencia (Spain).

This study was carried out to determine current levels of nitrate in vegetables marketed in the Region of Valencia (Spain) and to estimate the toxicological risk associated with their intake. A total of 533 samples of seven vegetable species were studied. Nitrate levels were derived from the Valencia Region monitoring programme carried out from 2009 to 2013 and food consumption levels were taken from the first Valencia Food Consumption Survey, conducted in 2010. The exposure was estimated using a probabilistic approach and two scenarios were assumed for left-censored data: the lower-bound scenario, in which unquantified results (below the limit of quantification) were set to zero and the upper-bound scenario, in which unquantified results were set to the limit of quantification value. The exposure of the Valencia consumers to nitrate through the consumption of vegetable products appears to be relatively low. In the adult population (16-95 years) the P99.9 was 3.13 mg kg-1 body weight day-1 and 3.15 mg kg-1 body weight day-1 in the lower bound and upper bound scenario, respectively. On the other hand, for young people (6-15 years) the P99.9 of the exposure was 4.20 mg kg-1 body weight day-1 and 4.40 mg kg-1 body weight day-1 in the lower bound and upper bound scenario, respectively. The risk characterisation indicates that, under the upper bound scenario, 0.79% of adults and 1.39% of young people can exceed the Acceptable Daily Intake of nitrate. This percentage could join the vegetable extreme consumers (such as vegetarians) of vegetables. Overall, the estimated exposures to nitrate from vegetables are unlikely to result in appreciable health risks.

Health risk assessment for exposure to nitrate in drinking water from village wells in Semarang, Indonesia.

The levels of nitrate in 52 drinking water wells in rural Central Java, Indonesia were evaluated in April 2014, and the results were used for a health risk assessment for the local populations by using probabilistic techniques. The concentrations of nitrate in drinking water had a range of 0.01-84 mg/L, a mean of 20 mg/L and a medium of 14 mg/L. Only two of the 52 samples exceeded the WHO guideline values of 50 mg/L for infant methaemoglobinaemia. The hazard quotient values as evaluated against the WHO guideline value at the 50 and 95 percentile points were HQ50 at 0.42 and HQ95 at 1.2, respectively. These indicated a low risk of infant methaemoglobinaemia for the whole population, but some risk for the sensitive portion of the population. The HQ50 and HQ95 values based on WHO acceptable daily intake dose for adult male and female were 0.35 and 1.0, respectively, indicating a generally a low level of risk. A risk characterisation linking birth defects to nitrate levels in water consumed during the first three months of pregnancy resulted in a HQ50/50 values of 1.5 and a HQ95/5 value of 65. These HQ values indicated an elevated risk for birth defects, in particular for the more sensitive population. A sanitation improvement program in the study area had a positive effect in reducing nitrate levels in wells and the corresponding risk for public health. For example, the birth defect HQ50/50 values for a subset of wells surveyed in both 2014 and 2015 was reduced from 1.1 to 0.71.

Development of a 4-NQO toxic equivalency factor (TEF) approach to enable a preliminary risk assessment of unknown genotoxic compounds detected by the Ames II test in UV/H2O2 water treatment samples.

An approach to enable a preliminary risk assessment of unknown genotoxic compounds formed by MP UV/H2O2 treatment of nitrate rich water, is described. Since the identity and concentration of specific genotoxic compounds is not established yet, a compound specific risk assessment cannot be performed. This limitation is circumvented by introducing a toxic equivalency factor, converting the concentration of unknown genotoxic compounds expressed by an Ames II test response into equivalent concentrations of 4-nitroquinoline oxide (4-NQO), to enable a preliminary risk assessment. Based on the obtained 4-NQO equivalent concentrations for the tested water samples and 4-NQO carcinogenicity data, an indication of the associated risk of the by MP UV/H2O2 treatment produced nitrated genotoxic compounds is obtained via the margin of exposure (MOE) approach. Based on a carcinogen study by Tang et al. (2004), a body weight of 70 kg and a drinking water consumption of 2 L per day, the 4-NQO equivalent concentration should not exceed 80 ng/L associated with a negligible risk. Application of this approach on samples from MP UV/H2O2 treated water of a full scale drinking water production facility, a 4-NQO equivalent concentration of 107 ng/L was established. These results indicate a safety concern in case this water would be distributed as drinking water without further post treatment.

Risk assessment of nitrate and petroleum-derived hydrocarbon addition on Contricriba weissflogii biomass, lifetime, and nutritional value.

Coastal diatoms are often exposed to both petroleum-derived hydrocarbon pollution and eutrophication. How these exposures influence on algal biomass, lifetime, and nutritional value are unknown. To examine a more accurate risk assessment of the pollutants on the role of diatoms in coastal ecosystem functions, Conticribra weissflogii was maintained at different concentrations of nitrate (N) and/or water-soluble fractions of No.0 diesel oil (WSF). Algal density, cell growth cycle, protein, chlorophyll a, superoxide dismutase (SOD) activity, and malonaldehyde (MDA) were determined for the assessment of algal biomass, lifetime, nutritional value, photosynthesis and respiration, antioxidant capacity, and lipid peroxidation, respectively.When N addition was combined with WSF pollution, the cell growth cycles were shortened by 27-44%; SOD activities were decreased by 1-64%; algal density, the concentrations of chlorophyll a, protein, and MDA were varied between 38 and 310%, 62 and 712%, 4 and 124%, and 19 and 233% of the values observed in N addition experiments, respectively. Coastal ecosystem functions were severely weakened by N and WSF additions, and the influence was increased in the order: N

Ecotoxicological assessment of the impact of nitrate (NO(3)(¯)) on the European endangered white-clawed crayfish Austropotamobius italicus (Faxon).

Field studies and laboratory experiments were conducted to assess the impact of elevated nitrate (NO(3)(¯)) concentrations on the European endangered white-clawed crayfish Austropotamobius italicus (Faxon) in the Henares River Basin (Central Spain), within an area that is vulnerable to nitrate pollution. Two sampling surveys were carried out in the summer of 2009 and 2011 to collect freshwater crayfish at eight sampling sites along this vulnerable area. The invasive read-swamp crayfish Procambarus clarkii (Girard) was the only collected crayfish species. Nitrate toxicity experiments however showed that A. italicus is one of the most tolerant species to nitrate toxicity. Although the food consumption was the most sensitive endpoint to nitrate toxicity (followed by the escape response and mortality), the no-observed-effect concentration (NOEC) for this endpoint after 14 days of exposure to nitrate was as high as 100mg NO(3)(¯)N/l, with some crayfish being still alive after fourteen days of exposure to a nominal nitrate concentration of 800mg NO(3)(-)N/l. Besides, a safe concentration of nitrate for A. italicus, along with its respective 95% confidence limits, were estimated to be 68.5 (22.4-187) mg (NO(3)(¯)N/l. Overall we conclude that elevated nitrate concentrations would not be responsible for the absence of white-clawed crayfish in the Henares River Basin. Other environmental factors, particularly the presence of P. clarkii and its fungal pathogen, would be major causes.