Snapshot of the pollution-driven metabolic and microbiota changes in Carassius gibelio from Bucharest leisure lakes.

In the last decades, increased intakes of contaminants and the habitats' destruction have produced drastic changes in the aquatic ecosystems. The environmental contaminants can accumulate in aquatic organisms, leading to the disturbance of the antioxidant/prooxidant balance in fish. In this context, we evaluated the level of organic, inorganic and microbiological pollutants in four leisure lakes (Chitila, Floreasca, Tei and Vacaresti) from Bucharest, the largest city of Romania, in order to compare their effects on hepatopancreas and gills metabolism and antioxidant defense mechanisms in Carassius gibelio, the most known and widespread freshwater fish in this country. The lowest level of oxidative stress was recorded in the case of fish collected from the Vacaresti lake, a protected wetland area where aquatic organisms live in wild environmental conditions. In contrast, significant oxidative changes were observed in the hepatopancreas and gills of fish from the Chitila, Floreasca and Tei lakes, such as reduced glutathione S-transferase activity and glutathione level, and increased degree of lipid peroxidation, being correlated with elevated levels of pesticides (such as 2,4'-methoxychlor) and Escherichia coli load in these organs. Although different patterns of pollutants' accumulation were observed, no important interindividual variations in cytosine methylation degree were determined. In conclusion, the presence and concentrations of metals, pesticides and antibiotics varied with the analyzed tissue and sampling site, and were correlated with changes in the cellular redox homeostasis, but without significantly affecting the epigenetic mechanisms.

Tissue Bioconcentration Pattern and Biotransformation of Per-Fluorooctanoic Acid (PFOA) in Cyprinus carpio (European Carp)-An Extensive In Vivo Study.

The perfluoroalkyl substances (PFAS) represent a persistent class of synthetic chemicals that spread in the environment as a result of industrialization. Due to their bioaccumulative and endocrine disruption implications, these chemicals can affect food quality and human health, respectively. In the present study, the bioconcentration and biotransformation of perfluorooctanoic acid (PFOA) in common carp (Cyprinus carpio) were evaluated in a biphasic system (exposure and depuration). Carp were continuously exposed, under laboratory conditions, to 10 (Experiment 1) and 100 (Experiment 2) µg/L PFOA for 14 weeks, followed by a wash out period of 3 weeks. Fish organs and tissues were collected at 8, 12, 14 weeks of exposure and at week 17, after the depuration period. The results obtained from the LC-MS/MS analysis showed the presence of PFOA in all studied organs. The highest values of PFOA were identified in the gallbladder (up to 2572 ng/g d.w.) in Experiment 1 and in the gallbladder (up to 18,640 ng/g d.w.) and kidneys (up to 13,581 ng/g d.w.) in Experiment 2. The average BCF varied between 13.4 and 158 L/Kg in Experiment 1 and between 5.97 and 80.3 L/Kg in Experiment 2. Four biotransformation products were identified and quantified in all organs, namely: PFBA, PFPeA, PFHxA, and PFHpA. PFBA was proven to be the dominant biotransformation product, with the highest values being determined after 8 weeks of exposure in the kidney, gallbladder, brain, liver, and gonads in both experiments. Because freshwater fish are an important food resource for the human diet, the present study showed the fishes' capacity to accumulate perfluoroalkyl substances and their metabolites. The study revealed the necessity of monitoring and risk studies of new and modern synthetic chemicals in aquatic resources.

Sensitivity of Pathogenic Bacteria Strains to Treated Mine Water.

Mine water as a result of meteoric and/or underground water's contact with tailings and underground workings could have an elevated content of metals associated with sulfate, often acidic, due to the bio-oxidation of sulfides. When entering aquatic ecosystems, the mine water can cause significant changes in the species' trophic levels, therefore a treatment is required to adjust the alkalinity and to remove the heavy metals and metalloids. The conventional mine water treatment removes metals, but in many cases it does not reduce the sulfate content. This paper aimed to predict the impact of conventionally treated mine water on the receiving river by assessing the genotoxic activity on an engineered Escherichia coli and by evaluating the toxic effects generated on two Gram-negative bacterial strains, Pseudomonas aeruginosa and Escherichia coli. Although the main chemical impact is the severe increases of calcium and sulfate concentrations, no significant genotoxic characteristics were detected on the Escherichia coli strain and on the cell-viability with a positive survival rate higher than 80%. Pseudomonas aeruginosa was more resistant than Escherichia coli in the presence of 1890 mg SO42-/L. This paper reveals different sensitivities and adaptabilities of pathogenic bacteria to high concentrations of sulfates in mine waters.

In Vivo Bioconcentration, Distribution and Metabolization of Benzophenone-3 (BP-3) by Cyprinus carpio (European Carp).

Organic UV-filters, such as oxybenzone (BP-3), have attracted researcher attention in recent years due to its capacity to interfere with the proper functioning of the endocrine system and its widespread presence in the aquatic environment. The aim of this study was to investigate the bioconcentration, distribution and metabolization of BP-3 in one of the most common fish species in Romania, namely Cyprinus carp (European carp). Exposure experiments were performed for 11 weeks using a BP-3 nominal concentration level of 100 µg/L. The BP-3 concentration level decreased over time and needed to be re-established daily. Biological samples (fish organs and tissues) from control and test were taken at t0 (before contamination) and at t3, t5, t8 and t11 weeks from the beginning of the experiment. From the third week, BP-3 was identified and quantified, in all organs, in concentration values ranging between 3.2 and 782 ng/g d.w., the highest concentration being detected in the intestinal content, followed by gonads (up to 468 ng/g d.w.) and skin (up to 453 ng/g d.w.). In the case of gill and liver, the BP-3 concentration increased in the first five weeks, and then decreased to 15 and 6 ng/g d.w., respectively, which could be explained by a fast BP-3 metabolization. During the exposure period, six metabolites were identified through LC-MS/MS, all of them known for their endocrine disruptor and toxic properties being higher than those of the parent compound. The study is important from an ecological perspective and also in view of human health concerns involving food quality.

Antioxidative Defense and Gut Microbial Changes under Pollution Stress in Carassius gibelio from Bucharest Lakes.

Fish are able to accumulate by ingestion various contaminants of aquatic environment, with negative consequences on their intestine, being continuously threatened worldwide by heavy metals, pesticides and antibiotics resulted from the human activities. Consequently, the health of other species can be affected by eating the contaminated fish meat. In this context, our study aimed to perform a comparison between the changes in intestine samples of Carassius gibelio individuals collected from different artificial lakes in Bucharest (Romania), used by people for leisure and fishing. The presence of various metals, pesticides and antibiotics in the gut of fish was assessed in order to correlate their accumulation with changes of antioxidative enzymes activities and microbiome. Our results showed that fish from Bucharest lakes designed for leisure (Chitila, Floreasca and Tei lakes) have an increased level of oxidative stress in intestine tissue, revealed by affected antioxidant enzymes activities and GSH levels, as well as the high degree of lipid peroxidation, compared to the fish from protected environment (Vacaresti Lake). Some heavy metals (Fe, Ni and Pb) and pesticides (aldrin and dieldrin) were in high amount in the gut of fish with modified antioxidative status. In conclusion, our study could improve the knowledge regarding the current state of urban aquatic pollution in order to impose several environmental health measures.

Oxidative Stress and Histopathological Changes in Gills and Kidneys of Cyprinus carpio following Exposure to Benzethonium Chloride, a Cationic Surfactant.

One cationic surfactant with a wide spectrum of microbiocidal activity is benzethonium chloride (BEC). Despite being widely used, the toxicity data on vertebrate organisms are limited. Therefore, we aimed to evaluate within this study the acute toxicity of BEC on the gills and kidneys of Cyprinus carpio (European carp). An alteration of the antioxidant enzymes activities (glutathione reductase, glutathione peroxidase and glutathione S-transferase) was noticed after 96 h of exposure, along with an elevation of lipid peroxidation and decreased concentration of reduced glutathione, which confirmed that BEC was able to induce toxicity to these tissues. These metabolic effects were correlated with unspecific structural changes observed in gills and kidneys, having moderate degree of severity (such as an increase of melanomacrophages aggregation incidence and cytoplasm vacuolation of goblet cells in collecting tubules) and generally being compatible with life for the exposure time studied. The most severe structural effects were observed in gills after 96 h, noticing a lamellar aneurysm, hemorrhages and lamellar epithelium disruption due to the blood vessels and pillar cells damages and increased blood flow inside the lamellae. By our research we can confirm the utility of biochemical and histological analyses in the fish organs as tools for monitoring the water quality and ecotoxicological potential of chemicals.

Degradation of Carbamazepine from Aqueous Solutions via TiO2-Assisted Photo Catalyze.

Photocatalytic degradation of carbamazepine (CBZ) from spiked aqueous solutions, via a UV/TiO2 system, was investigated, and the optimum photocatalyst type (P25 Degussa) and dose (500 mg/L), as well as irradiation time (45 min), were established. The degradation process kinetics was studied, and a degradation rate constant of 3.14 × 10-5 M min-1 was calculated for CBZ, using the Langmuir-Hinshelwood equation. Experiments performed in the presence of scavengers showed that the main reactive species involved in the degradation process are holes and free hydroxyl radicals; superoxide radicals also play a role in CBZ degradation. Eight transformation products of CBZ were identified, and a possible degradation pathway, consisting of four routes, was proposed. Toxicity and genotoxicity tests were also performed for both untreated and treated CBZ solutions, proving that the use of a UV/TiO2 system represents a suitable treatment approach for aqueous systems with CBZ content.

Bioavailability, Accumulation and Distribution of Toxic Metals (As, Cd, Ni and Pb) and Their Impact on Sinapis alba Plant Nutrient Metabolism.

This study presents the behavior of white mustard seedlings Sinapis alba grown for three months in laboratory polluted soil containing As, Cd, Ni and Pb. Four different experiments were performed in which As was combined with the other three toxic metals in different combinations (As, AsCd, AsCdNi, AsCdNiPb), keeping the same concentrations of As and Cd in all tests and following the national soil quality regulations. The effects of these metals were monitored by the analytical control of metal concentrations in soil and plants, bioavailability tests of mobile metal fractions using three different extracting solutions (DTPA + TEA + CaCl2-DTPA, DTPA + CaCl2-CAT, and CH3COONH4 + EDTA-EDTA) and calculation of bioaccumulation and translocation factors. Additionally, micro, and macro-nutrients both in soil and plant (root, stem, leaves, flowers and seeds) were analyzed in order to evaluate the impact of toxic metals on plant nutrient metabolism. Metals were significantly and differently accumulated in the plant tissues, especially under AsCdNi and AsCdNiPb treatments. Significant differences (p < 0.05) in the concentration of both As and Cd were highlighted. Translocation could be influenced by the presence of other toxic metals, such as Cd, but also of essential metals, through the competition and antagonism processes existing in plant tissues. Significantly, more Cd and Ni levels were detected in leaves and flowers. Cd was also detected in seeds above the WHO limit, but the results are not statistically significant (p > 0.05). The extraction of metallic nutrients (Zn, Cu, Mn, Ni, Mg, K, Fe, Ca, Cr) in the plant was not influenced by the presence of toxic metal combinations, on the contrary, their translocation was more efficient in the aerial parts of the plants. No phytotoxic effects were recorded during the exposure period. The most efficient methods of metal extraction from soil were for As-CAT; Cd-all methods; Pb and Ni-DTPA. The Pearson correlations (r) between applied extraction methods and metal detection in plants showed positive correlations for all toxic metals as follows: As-CAT > DTPA > EDTA, Cd-DTPA > CAT > EDTA, Ni-EDTA = DTPA > CAT, Pb-EDTA = DTPA = CAT). The results revealed that Sinapis alba has a good ability to accumulate the most bioavailable metals Cd and Ni, to stabilize As at the root level and to block Pb in soil.

Toxic Metals (As, Cd, Ni, Pb) Impact in the Most Common Medicinal Plant (Mentha piperita).

This study aimed to evaluate the behavior of Mentha piperita under Cd, Pb, Ni, and As soil contamination and their transfer from soil in plants as well as translocation in the roots/stems/leaves system compared with a control without metal addition. The mint seedlings were exposed for a three-month period using two metal mixtures in the same concentrations such as AsCd and AsCdNiPb (23.7 mg/kg As, 5 mg/kg Cd, 136 mg/kg Ni, and 95 mg/kg Pb). The results of metal concentration in plants showed that Cd, Ni, and Pb were accumulated in different parts of the plant, except for As. In plants organs, the order of metal accumulation was roots > stems > leaves. No significant impact on the growth, development, and chlorophyll content compared to the control was observed in the first month of exposure. After three months of exposure, phytotoxic effects occurred. Generally, the transfer coefficients and translocation factors values were less than 1, indicating that Mentha piperita immobilized the metals in root. The laboratory experiments highlighted that for a short period of time, Mentha piperita has the capacity to stabilize the metals at the root level and was a metal-tolerant plant when using a garden rich-substrate.

Risk screening of pharmaceutical compounds in Romanian aquatic environment.

The aquatic environment is under increased pressure by pharmaceutically active compounds (PhACs) due to anthropogenic activities. In spite of being found at very low concentrations (ng/L to µg/L) in the environment, PhACs represent a real danger to aquatic ecosystems due to their bioaccumulation and long-term effects. In this study, the presence in the aquatic environment of six non-steroidal anti-inflammatory drugs (ibuprofen, diclofenac, acetaminophen, naproxen, indomethacin, and ketoprofen), caffeine, and carbamazepine were monitored. Moreover, their aquatic risk and ecotoxicity by three biological models were evaluated. The monitoring studies performed in Romania showed that all studied PhACs were naturally present at concentrations >0.01 µg/L, pointing out the necessity to perform further toxicity tests for environmental risk assessment. The toxicity studies were carried out on aquatic organisms or bacteria and they indicated, for most of the tested PhACs, an insignificant or low toxicity effects: lethal concentrations (LC50) on fish Cyprinus carpio ranged from 42.60 mg/L to more than 100 mg/L; effective concentrations (EC50) on planktonic crustacean Daphnia magna ranged from 11.02 mg/L to more than 100 mg/L; inhibitory concentrations (IC50)/microbial toxic concentrations (MTC) on Vibrio fischeri and other bacterial strains ranged from 7.02 mg/L to more than 100 mg/L. The PhAC aquatic risk was assessed by using the ratio between measured environmental concentration (MEC) and predicted no effect concentration (PNEC) calculated for each type of organism. The average of quotient risks (RQs) revealed that the presence of these compounds in Romania's aquatic environment induced a lower or moderate aquatic risk.