Uptake, tissue distribution and elimination in a native fish species Astyanax eigenmanniorum exposed to inorganic mercury.

The increasing contamination of water bodies with mercury (Hg) raises concerns about the possible effects of this metal on native fish species. Our current understanding of its dynamics in fish organs remains limited. In this study, adult individuals of the native species Astyanax eigenmanniorum were exposed to three environmentally relevant HgCl2 concentrations (5, 100, and 170 µg L-1) for 96 h. To evaluate total Hg (THg) elimination, new individuals were exposed to 100 µg L-1 of HgCl2 (96 h), and at the end of the exposure period, half of the fish were placed in tanks with clean water for 168 h. In both assays, the organs were removed, and THg levels were measured using ICP-MS. The uptake of IHg in A. eigenmanniorum showed a differential accumulation in the organs. Gills, intestine, and brain were the tissues with the highest THg levels. Finally, no elimination of THg in the water was observed, but intestine and gills significantly removed the THg accumulated. Probably a Hg redistribution through the tissues could take place.

Trace element concentrations in six fish species from freshwater lentic environments and evaluation of possible health risks according to international standards of consumption.

Multi-element concentrations (Al, As, Cd, Cr, Hg, Ni, Pb, Se, and Sr) were analyzed in the muscle of six fish species (Hoplias malabaricus, Oligosarcus jenynsii, Rhamdia quelen, Bryconamericus iheringii, Astyanax fasciatus, and Odontesthes bonariensis) with different diets and habits from the Río Tercero Reservoir (RTR) in Córdoba, Argentina, during the wet and dry seasons. Besides, potential human health risks, associated with the consumption of these elements, have been assessed considering the average daily intake (EDI) in children and adults. Additionally, the target hazard quotient (THQ) and carcinogenic risk (CR) were evaluated taking into account the intake by the general population, fishermen, and consumption frequency recommended by the American Heart Association (AHA), the maximum scientific reference in cardiology in the USA and worldwide. All species presented quantifiable values in muscle for all the analyzed elements (Al, As, Cr, Hg, Ni, Se, and Sr), except for Cd and Pb, being Al and Sr the most accumulated elements in all species in both seasons. The consumption of edible muscles of the species studied in this reservoir represents a toxicological risk to humans. Mercury and As were the main elements that presented a health risk through the consumption of fish. Their concentrations in most fish species were above the maximum daily allowable concentrations, and THQ values were several times greater than 1. In addition, according to AHA recommendations, the cancer risk caused by As was greater than the acceptable value of 10-4 in all species studied, and in both seasons, with the exception of A. fasciatus, in the rainy season. These results indicate that the consumption of fish from the RTR exposes the inhabitants to possible health risks, especially when considering the consumption frequency recommended by the AHA. Therefore, fish intake from this reservoir should be limited to minimize potential risks to the health of consumers. Finally, the results of this study are useful for controlling pollution and developing preventive and palliative policies to protect populations in contact not only with the reservoir but also with other areas of the world with similar conditions.

Potential human health risks from metals and As via Odontesthes bonariensis consumption and ecological risk assessments in a eutrophic lake.

The concentration of Al, Cr, Fe, Mn, Ni, Cu, Zn, Hg, Sr, Mo, Ag, Cd, Pb and As was analyzed in water, sediment, and muscle of Odontesthes bonariensis from the eutrophic San Roque Lake (Córdoba-Argentina). The monitoring campaign was performed during the wet, dry and intermediate season. The concentration of Cr, Fe, Pb, Zn, Al and Cd in water exceeded the limits considered as hazardous for aquatic life. The highest metal concentrations were observed in sediment, intermediate concentrations, in fish muscle, and the lowest in water, with the exception of Cr, Zn, As and Hg, which were the highest in fish muscle. Potential ecological risk analysis of heavy metal concentrations in sediment indicated that the San Roque Lake posed a low ecological risk in all sampling periods. The target hazard quotients (THQs) and carcinogenic risk (CR) for individual metals showed that As in muscle was particularly hazardous, posing a potential risk for fishermen and the general population during all sampling periods. Hg poses a potential risk for fishermen only in the intermediate season. It is important to highlight that none of these two elements exceeded the limits considered as hazardous for aquatic life in water and sediment. This result proves the importance of performing measurements of contaminants, in both abiotic and biotic compartments, to assess the quality of food resources. These results suggest that the consumption of this fish species from this reservoir is not completely safe for human health.

Effects of carbamazepine on cortisol levels and behavioral responses to stress in the fish Jenynsia multidentata.

Carbamazepine (CBZ) is an anticonvulsant drug, prescribed worldwide for the treatment of epilepsy, bipolar disorder and trigeminal neuralgia, which has been frequently detected in aquatic environments. The objective of this study was to analyze if CBZ modifies scototaxis and shoaling behaviors and/or whole-body cortisol levels of the one-sided livebearing fish Jenynsia multidentata under stress condition. Female adults of J. multidentata were exposed to 0, 10, 50 and 200µgCBZ/L during 14days. After CBZ exposure, fish were subjected to restraint stress during 15min. Control animals were not exposed to CBZ or stress. In the light/dark preference test (scototaxis), the individuals under acute restraint stress (without CBZ) exhibited a significant increase in the mean speed and in the time spent both in the light compartment and in the bottom of the tank with respect to controls. They also showed a tendency to stay longer frozen in the light compartment. Fish exposed to 10 and 50µgCBZ/L showed a significant reduction in mean speed compared to stressed fish without CBZ. A reduction in the time spent in the bottom of the tank was also observed in fish exposed to 10µgCBZ/L. Fish exposed to 200µgCBZ/L showed a decreasing tendency in all behavioral endpoints (time spent in the light compartment, mean speed, time spent at the bottom and freezing) in comparison to stressed fish not exposed to CBZ. Considering whole-body cortisol results, fish under acute restraint stress (without CBZ) significantly increased their hormone levels with respect to the control group, while fish exposed to CBZ and acute restraint stress, significantly decreased their whole-body cortisol levels. There were no significant changes in shoaling behavior due to either stress or CBZ exposure and no significant differences in whole-body cortisol levels between experimental groups. Considering that the light/dark and shoaling tests measure different stress response behaviors regulated by different neuroendocrine systems, these results could indicate that CBZ has a differential effect on fish behavioral stress response and cortisol levels, depending on the behavioral test used and stressor applied.

Occurrence and bioaccumulation of pharmaceuticals in a fish species inhabiting the Suquía River basin (Córdoba, Argentina).

In South America, there is a lack of data concerning the occurrence and levels of pharmaceuticals in main rivers as well as their negative effects on the biota. Here we report the occurrence as well as the spatial and temporal variations of some common prescribed pharmaceuticals in the Suquía River basin (Córdoba, Argentina). We also report the bioconcentration of two of them in Gambusia affinis, a widely distributed fish species inhabiting the river basin. The influence of the wastewater treatment plant of Córdoba City was critical (up to 70 km downstream). Among 15 compounds analyzed, atenolol, carbamazepine and diclofenac were the most frequently detected (reaching sub µg L(-1) levels), showing different distribution patterns. Bioconcentration of atenolol and carbamazepine was studied under laboratory controlled conditions. Estimated bioconcentration factors (BCFs) were: 0.13 and 0.08 L kg(-1) upon exposure to 100 and 1,000 µg L(-1) atenolol in water, respectively; while BCFs were 0.7 and 0.9 L kg(-1) when exposed to 10 and 100 µg L(-1) carbamazepine, respectively. To the extent of our knowledge, this is the first report on pharmaceuticals in superficial waters of Argentina as well as the first report on the bioaccumulation of atenolol in whole body fish.

Reproductive impairment of a viviparous fish species inhabiting a freshwater system with anthropogenic impact.

The potential threat to animal reproduction by contaminated freshwater systems posed the necessity to identify and develop bioindicators and biomarkers to be used for screening and evaluation of the effects in organisms. The main goal of this work was to determine, through histological analyses and changes in gonopodium morphology, whether a freshwater system polluted by anthropogenic activities-sewage, agricultural, and industrial-could cause alterations at the organ level. We also propose the live-bearing fish, Jenynsia multidentata, as a species suitable to study the effects of contaminated aquatic environments. We compared male fish sampled at two different stations in Suquía River basin (Córdoba, Argentina), both differing in degree of pollution, through liver and testis histology and gonopodial morphometric parameters. The water quality, based on the physicochemical characteristics of the studied stations, varied markedly with a decrease in water quality at the downstream site (station 2). At the highest polluted area, detrimental effects on liver and testis were evidenced on histological analysis. Male individuals from station 2 also presented noticeable structural changes of the anal fin, such as a straight gonopodium and abnormal tip area. The present results demonstrate that a freshwater system polluted by the impacts of anthropogenic activities has detrimental effects to J. multidentata. The alterations registered in individuals from the polluted station indicate an impairment of male reproductive performance and imply a risk for other live-bearing species as well as the entire biodiversity. We consider J. multidentata a sentinel species that is useful to evaluate the potential risk present in the studied basin not only to itself but to other species as well.

Histopathological changes in the gills and liver of Prochilodus lineatus from the Salado River basin (Santa Fe, Argentina).

This study evaluated the histopathological changes in gills and liver of Prochilodus lineatus inhabiting the Salado River basin. Fish were collected in four different sampling stations. The histological lesions in the tissues were examined under light microscopy and evaluated with quantitative analyses. The morphometric analysis of the gills showed a significant shortening of secondary lamellae and a lower percentage of area for gas exchange in fish from station 1 (an urban area, located near the mouth of the Salado River) in comparison with fish gills from the reference site (station 4, a relatively pristine area). Moreover, a significantly higher area occupied with necrotic foci and the occurrence of an important inflammatory response were observed in fish liver of station 1 than the samples caught from other stations. Thus, histopathological evidences showed differences among sites, which could be related to different environmental conditions.

Attenuating effects of natural organic matter on microcystin toxicity in zebra fish (Danio rerio) embryos -- benefits and costs of microcystin detoxication.

To contribute to the understanding of joined factors in the environment, impact of pure microcystins (-RR and -LF) on zebra fish (Danio rerio) embryos were investigated individually and in combination with a natural organic matter (NOM). The applied NOM was a reverse osmosis isolate from Lake Schwarzer See (i.e., Black Lake, BL-NOM). Teratogenic effects were evaluated through changes in embryonic development within 48 h of exposure. Detoxication activities were assessed by the activities of phase II biotransformation enzymes, soluble and microsomal glutathione S-transferase (s, mGST). Oxidative stress was assessed by determining both the production of hydrogen peroxide and by analyzing the activities of the antioxidative enzymes, guajacol peroxidase (POD), catalase (CAT), glutathione peroxidase (GPx), and the glutathione restoring enzyme glutathione reductase (GR). Energetic costs were evaluated by determining contents of fat, carbohydrates, and proteins in both exposed and control embryos. BL-NOM attenuated toxic effects of MC-LF and MC-RR verified by less pronounced teratological effects within 24 h, in particular, as well as less rise in the activity of s-GST, when compared with embryos exposed to either pure toxins or in combination with organic matter. BL-NOM also diminished oxidative effects caused by MC-LF; however, it failed to attenuate oxidative stress caused by MC-RR. Content of lipids was significantly reduced in exposed embryos following a trend similar to that obtained with teratological and enzymatic assays confirming the attenuating effect of BL-NOM. Physiological responses to microcystins and NOM required energetic costs, which were compensated to the expense of the energy resources of the yolk, which in turn might affect the normal development of embryos.

Differential detoxification and antioxidant response in diverse organs of Corydoras paleatus experimentally exposed to microcystin-RR.

We assessed changes in activities of both detoxification and antioxidant enzymes as well as lipid peroxidation levels in liver, gill, intestine and brain of Corydoras paleatus exposed to dissolved microcystin-RR (MC-RR). Fish were captured at an unpolluted area, transported to the laboratory, and acclimated previous to experiments. Exposures were carried out using MC-RR at 0.5, 2, 5 and 10 microg L(-1). After exposures for 24h, fish were sacrificed and dissected separating liver, gills, intestine and brain of each fish. Organs were used for enzyme extractions, evaluating both antioxidant and detoxification systems through the assay of glutathione reductase (GR), guaiacol peroxidase (POD), glutathione peroxidase (GPx), catalase (CAT) as well as glutathione S-transferase (GST). Additionally, thiobarbaturic acid (TBA) method was used to evaluate the peroxidation of lipids (LPO). GST was inhibited in all studied organs at most MC-RR concentrations used. Activities of GR, POD and GPx were enhanced in liver at 2 microg L(-1), but inhibited in gills at all tested concentrations. CAT activity was enhanced in liver at all studied concentrations. Antioxidant response in liver is activated at low toxin concentrations, followed by a drop at the highest MC-RR levels. On the contrary, detoxification activity is inhibited in liver and brain in a dose-dependent way. On the other hand, MC-RR (>or=2 microg L(-1)) induced LPO in brain of exposed fish, but not in other organs. This finding becomes to this organ in one of the most severely affected. Results show that gills are also very affected, since both antioxidant and detoxification systems were inhibited in this tissue. Thus, inhibition of these defense systems could increase the uptake of different toxics through gills of fish exposed to dissolved MC-RR, leading to an increased health risk for fish. The different response observed on diverse organs exposed to MC-RR might be related to the uptake route as well as on biotransformation and bioaccumulation capabilities of different studied organs.

Recovery of Escherichia coli in fresh water fish, Jenynsia multidentata and Bryconamericus iheringi.

Escherichia coli concentration was determined in digestive tract and muscle of Jenynsia multidentata and Bryconamericus iheringi through bioassays. Field experiments were also conducted with J. multidentata collected in the Suquía River, Córdoba, Argentina. E. coli was quantified by the most probable number, using lauryl sulphate tryptose broth with 4-methylumbelliferyl-beta-D-glucuronide. For bioassays, E. coli concentrations 10(2), 10(3), 10(4), 10(5), 10(6)CFU/ml were introduced in aquarium water. E. coli was recovered from the digestive tracts of J. multidentata and B. iheringi in all the concentrations assayed. Bacterial critical load in water for the recovery of bacteria from muscle, was 10(3)CFU/ml for both species. The regression analysis between E. coli loads in water and those found in digestive tract and muscle showed a positive linear relationship for J. multidentata and B. iheringi. The same relation was observed between the concentration of bacteria in digestive tract and muscle in both species. In field experiments, E. coli was recovered from digestive tract and muscle of J. multidentata. The presence of E. coli in the studied fish suggests that they can carry bacteria to non-polluted waters. However, further studies are necessary to evaluate its significance for public and environmental health.