Biomarkers of a native fish species (Cnesterodon decemmaculatus) application to the water toxicity assessment of a peri-urban polluted river of Argentina

Environmental monitoring by means of biomarker parameters assessed in different species is a useful tool. It has the advantage of providing a quantitative response as well as valuable information of ecological relevance on the chronic adverse effects caused by water pollution. The aim of this study was to assess the response of biochemical and physiological parameters of Cnesterodon decemmaculatus, a native teleost, simultaneously caught in two sites of Reconquista river, a highly polluted peri-urban river. This study compared the measured parameters with that of specimens of the same species captured in an unpolluted body water, and correlated the detected changes with the physicochemical profile of the water at each site. A comparison was made of selected parameters of gill, brain and liver and of somatic indices of fish collected from polluted and reference sites. The main parameters whose changes allowed to discriminate between sampling sites were gill (Na(+),K(+))-ATPase, brain acetylcholinesterase (AChE) and liver aminotransaminases activities; tissues' protein content and liver somatic index (LSI) were also sensitive biomarkers in brain and liver, respectively. The results showed that the response of the measured biomarkers allowed for the differentiation of sampling sites according to their water quality and confirmed that Cnesterodon decemmaculatus may be a useful test organism for the biomonitoring of freshwater environments. In addition, the simultaneous measurement of the physicochemical parameters of the water samples showed a good correspondence between the biomarkers responses and the environmental chemical stress conditions.     

Biomarkers assessment in juvenile Cyprinus carpio exposed to waterborne cadmium

The impact of long-term exposure to waterborne cadmium (Cd) on Cyprinus carpio was evaluated through changes of selected parameters considered as biomarkers of toxicity. Fish were exposed to 1.6 mg l(-1) Cd for 14 days and then transferred to Cd-free water for 19 days. The measured parameters were gill ATPases, brain acetylcholinesterase (AchE), liver glutamate oxaloacetate (GOT) and glutamate pyruvate (GPT) transaminases, muscle water content, and protein content of liver, gills and brain. Condition factor and liver somatic index were also calculated. Branchial ATPase activities were impaired in a dissimilar way: the (Na(+),K(+))-ATPases were inhibited by approximately 30%, while the Mg(2+)-ATPase was significantly activated by 70%. Brain AchE showed no changes after Cd exposure. Both liver GOT and GPT activities were increased by the metal by 63 and 98%. Water content of the skeletal muscle showed no significant alterations. After the 19-day recovery phase, changes in the Mg(2+)-ATPase and GPT were reversed to values similar to controls, but the Cd exposure resulted in an irreversible alteration in GOT activity. Results indicate that the sublethal Cd concentrations are stressful to carp, particularly with reference to branchial enzymes which may disrupt the osmotic and ionic balance of the animals.     

Sub-chronic effect of neem based pesticide (Vepacide) on acetylcholinesterase and ATPases in rat.

Acetylcholinesterases (AChE), Na(+)-K+, Mg2+ and Ca(2+)-ATPases were monitored in rat brain when treated orally with 80, 160 and 320 mg/kg of Vepacide, an active ingredient from neem seed oil, daily for 90 days. Brain AChE, Na(+)-K+ and Ca(2+)-ATPases were inhibited whereas Mg(2+)-ATPase levels were enhanced in both the sexes after 45 and 90 days of treatment. The relative sensitivities of these ATPases to Vepacide indicated that Ca(2+)-ATPase being more sensitive than Na(+)-K(+)-ATPase in both the sexes. The magnitude of Ca(2+)-ATPase inhibited by this compound was higher than that of brain AChE. It appears to be sexual dimorphism in the alterations of brain AChE, Na(+)-K+ and Mg(2+)-ATPases by Vepacide with females being significant when compared with males. After 28 days of post treatment the alterations observed were approached to those of controls both in male and female rats showing reversal of the toxicity. These results indicated that the ATPases were potently inhibited by Vepacide and seemed to be its precise target among the enzyme studied. This can be used as biochemical marker of exposure to this neem derived product.     

Biochemical alterations in euryhaline fish, Oreochromis mossambicus exposed to sub-lethal concentrations of an organophosphorus insecticide, monocrotophos

The euryhaline fish, Oreochromis mossambicus was exposed to sub-lethal concentration (1.15 mg l(-1)) of a organophosphorus insecticide, monocrotophos (MCP) for 30 days and allowed to recover for seven days. Alanine aminotransferase (ALAT), aspartate aminotransferase (AAT), acid phosphatase (AcP), alkaline phosphatase (ALP), glycogen, lactate dehydrogenase (LDH), Reduced glutathione (GSH), gluthathione-S-transferase (GST) and acetylcholinesterase (AChE), were assayed in plasma and different tissues at regular intervals of day -3, -7, -15, -30 and after recovery period of seven days. The ALAT and AAT activities were increased in plasma and kidney, where as liver and gill showed decrease. Increase in AcP and ALP activities were observed in plasma, gill and kidney, and reduction of 42% and 50% was observed in liver. Glycogen was depleted in plasma, liver and gill indicates of typical stress related response of the fish with pesticide. LDH activity was decreased in liver and muscle, indicating tissue damage and muscular harm, but a significant increase in LDH activity in gill and brain was observed. Depletion in GSH activity was observed in all the tissues, there by enhancing the lipid peroxidation resulting in cell damage. The induction in hepatic GST levels indicates the protection against the toxicity of xenobiotic-induced lipid peroxidation. There was a significant recovery in all the above biochemical parameters studied in plasma and different tissues, after seven days recovery period. These results revealed that MCP affects the intermediary metabolism of O. mossambicus and that the assayed enzymes can work as good biomarkers of organophosphorus contamination.     

Toxicological response of silver catfish (Rhamdia quelen) after acute exposure to a commercial insecticide containing thiamethoxam

Abstract

This study assessed the hematological, enzymatic and osmoregulatory responses of silver catfish (Rhamdia quelen) exposed to sublethal concentrations (1.125 and 3.750?µg/L) of a commercial thiamethoxam-containing insecticide used on rice crops. Groups of 6 fish per tank (in triplicate, n?=?3, total 54 fish) were exposed for up to 96?h to different concentrations of the compound. After this period, fish were placed in clean water for 48?h. Two fish from each tank (6 per treatment) that had been exposed to the insecticide for 24?h were anesthetized with eugenol and blood was collected to evaluate hematological and biochemical parameters. Blood, liver and muscle were collected for determination of metabolic parameters, plasma cortisol, Cl^-, Na⁺ and K⁺ levels and H⁺-ATPase and Na⁺/K⁺-ATPase activity in the gill. H⁺-ATPase activity was higher in fish exposed to 1.125?µg/L insecticide at 24?h compared to control (0.0?µg/L). Differences in cortisol levels were evidenced throughout the experimental period. These results indicated that exposure to the insecticide changed the hematological, biochemical and metabolic profile of the animals, suggesting concern about environmental safety. Therefore, we discourage the use of this pesticide in areas that come into contact with water bodies inhabited by fish.     

The significance of water ionic strength on aluminium toxicity in brown trout (Salmo trutta L.)

The toxicity of aluminium to fish is related to interactions between aluminium and the gill surface. We investigated the possible effect of water ionic strength on this interaction. The mortality of brown trout (Salmo trutta L.) exposed to three different degrees of Al polymerisation was compared in water with increased ionic strength (mean 7.31 x 10(-4) M) after additions of the base cations Ca2+, Mg2+, Na+ or K+, and in water with no such addition (mean ionic strength 5.58 x 10(-4) M). Only a very slight ameliorating effect of increased ionic strength was observed, while the degree of Al polymerisation was of major importance in fish mortality. In addition, it was observed that smaller fish survived the Al exposures for a longer time than larger fish. We hypothesise that this is because larger fish are more susceptible to hypoxia than smaller fish.     

Biochemical changes produced by beta- and gamma-hexachlorocyclohexane isomers in albino rats

Administration of beta- and gamma-isomers of hexachlorocyclohexane (HCH) at 800 ppm dietary level for 2 weeks to albino rats produced noticeable hepatocellular damage as indicated by elevations in serum aminotransferases and decreases in hepatic soluble enzymes. Although serum total LDH activity was not altered, the LD5 isoenzyme was proportionately higher in the HCH isomers treated animals. Treatment of rats with beta- and gamma-isomers of HCH increased the hepatic glucose-6-phosphate dehydrogenase and aldolase activities suggesting a higher rate of glucose oxidation. Liver glucose-6-phosphatase activity was decreased in these animals indicating inactivation of gluconeogenesis in liver. Dietary beta- and gamma-HCH decreased the liver mitochondrial DNP/Mg++/Ca++-activated ATPases thus affecting the energy metabolism. An unaltered ratio of DNP/Mg++-ATPase, a study of swelling pattern of hepatic mitochondria, and NAD+ permeability test suggested the maintenance of structural integrity of mitochondrial membrane in these pesticide fed animals. Liver microsomal Na+,K+-ATPases were lower in these animals.     

The influence of lindane and lindane metabolites on microsomal and mitochondrial ATPase in vitro

In vitro investigations of the influence of lindane and its metabolites were performed on microsomal and mitochondrial ATPases from liver, kidney and brain of rat and mouse. The microsomal Na+-K+-ATPases in rat liver were inhibited by the tested substances. An increase of activity was observed only with 2.5 X 10(-5) M gamma-HCH. Effects on the microsomal Na+-K+-ATPase from kidney and brain of rat were also indicated. The mitochondrial enzyme in rat liver was stimulated by all the compounds tested at concentrations of 10(-4) M - 10(-2) M. The effects on mitochondrial enzymes from kidney and brain varied in dependence on the tested substances. In the microsomes and mitochondria of mouse an influence on the Na+-K+-ATPases similar to the effects on the preparations from organs of rat was evident.     

Chemical contamination of the Rybinsk Reservoir, northwest Russia: relationship between liver polychlorinated biphenyls (PCB) content and health indicators in bream (Abramis brama)

The Rybinsk Reservoir (Russia) is the largest artificial waterbody in Europe (4550 km2) and provides drinking water for population of the cities located along the coast line. Industrialization in Cherepovets at the northeastern portion of the reservoir, including one of the largest metallurgical facilities in Europe, has resulted in chemical contamination of the reservoir. The extent of polychlorinated biphenyls (PCB) contamination in bream liver, a common fish species, taken from six locations in the Rybinsk Reservoir and Volga River, and biochemical and morphometric biomarkers of fish health were investigated. Liver PCB concentrations ranged from non-detected to 3.4 microg/g wet wt of liver, with the greatest concentrations found in fish taken near the industrialized area in Sheksna Reach of Rybinsk Reservoir. The source of the bream contamination is the PCB pollution of bottom organisms and sediments conditioned with industrialization facilities of Cherepovets. The patterns of the PCB congeners in the livers of bream taken near Cherepovets were similar at all of the stations that were sampled around the reservoir and Volga River. Among the common fish health biomarkers used only liver total ChE activity and liver-somatic index in bream near Cherepovets can reflect environmental pollution. Other morphometric (FCF, Clark's condition factors, and spleen-somatic index) and biochemical (protein content and acetylcholinesterase activity in the brain) biomarkers related with fish health varied among locations, but were not correlated to the concentrations of PCBs in the bream livers.     

Multiple biomarkers of pollution effects in Solea solea fish on the Tunisia coastline

This field study investigates the morphological indices (condition index, hepatosomatic index) and biochemical (catalase (CAT), glutathione S-transferase (GST), acetylcholinesterase (AChE), metallothionein (MT), lipid peroxidation) parameters in liver, gills and kidney of common sole (Solea solea) originating from different sites of the Tunisian coast area impacted by different anthropogenic activities. Differences among sites and tissues for AChE, GST, CAT, MT and TBARS were found and possibly related to known sources of domestic and industrial discharges in the studied sites. Liver, gills and kidney CAT, liver and kidney MT and brain AChE were key biomarkers to discriminate fish of different sites. So, we suggest using these biomarkers in future biomonitoring.     

Alterations in liver marker enzymes and recovery ability of cyprinids as indicators of aquatic pollution

Information on the detection of pesticides in fresh water Lakes of Kashmir (Dal and Mansbal) through GC–MS technique is scarce, and also the recovery in biochemical parameters (AST, ALT and ALP) of fish after transferring them to clean media has not been reported yet. The water samples were collected from three sites and analyzed for their pesticide profile by dispersive liquid–liquid micro extraction (DLLME) followed by GC–MS. Influence of pesticides on liver marker enzymes of Cyprinus carpio and Carassius carassius was also investigated. The results obtained showed the presence of three main pesticides viz. chlorpyrifros, dimethoate and dichlorvos in waters of Dal Lake whereas no pesticide was detected in waters of Mansbal. The higher values for AST, ALT and ALP activities and decrease in protein content were obtained in the samples from the Dal Lake compared with those from Mansbal Lake (p < 0.05). These data when compared with the values found in C. carpio and C. carassius from both the Dal Lake and Mansbal Lake kept under laboratory conditions after 15, 30, 45 and 60 days of maintenance in clean media found that during depuration, all the enzyme activities came down significantly (p < 0.05) indicating the compensatory response by the fish against the pesticide stress. Therefore, these parameters could be used as indicators of pesticide pollution in aquatic organisms and were recommended for environmental monitoring for investigating the mechanism involved in the recovery pattern.

     

First evidence of polybrominated diphenyl ether (flame retardants) effects in feral barbel from the Ebro River basin (NE, Spain)

Polybrominated diphenyl ethers (PBDEs) are chemicals of environmental concern due to their lipophilic, persistent and bioaccumulable characteristics as well as for their potential endocrine disrupting role. Former studies carried out in a tributary of the Cinca river (Ebro basin, NE Spain) revealed high levels of PBDEs in fish due to the discharges of effluents rich in PBDEs coming from a nearby industrial park in Barbastro. In this study, several biomarkers of pollutants exposure were measured in barbel, Barbus graellsii, before (upstream) and after (downstream) the main industrial site (Barbastro) in the Vero river. The results evidenced an enhanced hepatic phase I and II metabolism (measured as reductases, glutathione S transferase and uridinediphospho-glucuronosyltransferase), and of the antioxidant enzyme glutathione peroxidase. Conversely, fishes collected from downstream reaches had their phase I CYP1A dependent ethoxyresorufin O-deethylase, antioxidant diaphorase and brain cholinesterase activities depleted. In addition, the histological study of the liver and kidney of these fish evidenced an increase of the number and size of macrophage aggregates in most individuals collected downstream. Bivariate correlated analyses showed that the above mentioned biomarkers were correlated to measured PBDE congeners, thus indicating that the observed biological effects were unlikely to be related to other environmental factors than PBDEs. Overall, the measured biochemical and histological markers provide new evidence that in field exposed fish, PBDEs levels were associated with high activities of phase I and II metabolic enzymes, oxidative stress in liver, neurotoxicity in brain and histopathological effects in both liver and kidney.     

Effects of environmentally relevant concentrations of metallic compounds on the flatfish Scophthalmus maximus: biomarkers of neurotoxicity,oxidative stress and metabolism

Flatfish species, such as the turbot (Scophthalmus maximus), are common targets for toxic effects, since they are exposed through the food chain (ingestion of contaminated preys) and are in direct contact with the waterborne contaminant and sediments. Furthermore, these fish species live in close proximity to interstitial water that frequently dissolves high amounts of contaminants, including metals. Despite this significant set of characteristics, the present knowledge concerning flatfish contamination and toxicity by metals is still scarce. To attain the objective of assessing the effects of metals on a flatfish species, S. maximus specimens were chronically exposed to lead, copper and zinc, at ecologically relevant concentrations, and biochemical (oxidative stress: catalase and glutathione S-transferases activities, and lipid peroxidation; neurotoxicity: cholinesterase activity) parameters were assessed on selected tissues (gills and liver). Copper had no significant effects on all tested parameters; lead was causative of significant increases in liver GSTs activities and also in lipoperoxidation of gill tissue; exposure to zinc caused a significant increase in catalase activity of gill tissue. None of the tested metals elicited noteworthy effects in terms of neurotoxicity. The obtained results showed that only the metal lead is of some environmental importance, since it was able to cause deleterious modifications of oxidative nature at relevant concentrations.     

Development and validation of a terrestrial biotic ligand model predicting the effect of cobalt on root growth of barley (Hordeum vulgare)

A Biotic Ligand Model was developed predicting the effect of cobalt on root growth of barley (Hordeum vulgare) in nutrient solutions. The extent to which Ca(2+), Mg(2+), Na(+), K(+) ions and pH independently affect cobalt toxicity to barley was studied. With increasing activities of Mg(2+), and to a lesser extent also K(+), the 4-d EC50(Co2+) increased linearly, while Ca(2+), Na(+) and H(+) activities did not affect Co(2+) toxicity. Stability constants for the binding of Co(2+), Mg(2+) and K(+) to the biotic ligand were obtained: logK(CoBL)=5.14, logK(MgBL)=3.86 and logK(KBL)=2.50. Limited validation of the model with one standard artificial soil and one standard field soil showed that the 4-d EC50(Co2+) could only be predicted within a factor of four from the observed values, indicating further refinement of the BLM is needed.     

Multiple biomarkers responses in Prochilodus lineatus allowed assessing changes in the water quality of Salado River basin (Santa Fe, Argentina)

This field study assessed water quality of Salado River basin by using a set of biomarkers in the fish Prochilodus lineatus. Multiple biomarkers were measured, including morphological indexes (condition factor, liver somatic index), hematological (red and white blood cells) and biochemical (glucose, total protein and cholinesterase activity) parameters. Besides, detoxication and oxidative stress markers (antioxidant enzymes, lipid peroxidation) were measured in liver, gills and kidney. Despite water quality assessment did not show marked differences among sites, biomarkers responses indicate that fish are living under stressful environmental conditions. According to multivariate analysis glucose, glutathione S-transferase activity, lipid peroxidation levels and the count of white blood cells are key biomarkers to contribute to discrimination of sites. So, we suggest use those biomarkers in future monitoring of freshwater aquatic systems.     

Responses of antioxidant status and Na+–K+-ATPase activity in gill of rainbow trout,Oncorhynchus mykiss,chronically treated with carbamazepine

In recent years, chemical pollution by the residual pharmaceuticals has been increasingly important issue due to its widely present in the aquatic environment. However, the toxicological effects of residual pharmaceuticals on fish have not been adequately researched. The aim of this work is to investigate the toxic effect of CBZ, an anticonvulsant drug commonly present in aquatic environment, on antioxidant status and Na⁺–K⁺-ATPase in gill of rainbow trout exposed to sublethal CBZ (1.0 μg L⁻¹, 0.2 mg L⁻¹ and 2.0 mg L⁻¹) for 7, 21 and 42 d. After prolonged exposure of CBZ at higher test concentration (0.2 or 2.0 mg L⁻¹), oxidative stress was apparent as reflected by the significant higher LPO and CP levels in fish gill, as well as the significant inhibition of antioxidant enzymes activities including SOD, CAT, GR and GPx. Besides, reduced glutathione level and Na⁺–K⁺-ATPase activity were significantly lower than those of the control after 42 d of exposure to CBZ at higher test concentration (0.2 or 2.0 mg L⁻¹). The results of this study indicate that chronic exposure of CBZ has altered multiple physiological indices in fish gill; however, before those parameters are used as special biomarkers for monitoring residual pharmaceuticals in aquatic environment, more detailed experiments in laboratory need to be performed in the future.     

Histopathological biomarkers in gills and liver of Oreochromis niloticus from polluted wetland environments, Saudi Arabia

Fish live in direct contact with their immediate external environment and, therefore, are highly vulnerable to aquatic pollutants. In this study, Oreochromis niloticus were caught at three different sites in Al-Hassa irrigation channels, namely Al-Jawhariya, Um-Sabah and Al-Khadoud. The histological changes in gills and liver were detected microscopically and evaluated with semi-quantitative analyses. Also, heavy metals have been determined in the water samples in these sites. Results showed that all sites were polluted by different kinds of heavy metals. Cd and Pb were mostly detected at concentrations above the WHO reference values. Meanwhile, various histopathological abnormalities were observed in gills and liver of fish specimens. In the gill filaments, cell proliferation, lamellar cell hyperplasia, lamellar fusion, lifting of the respiratory epithelium, and the presence of aneurysmal areas were observed. In the liver, there was vacuolization of the hepatocytes, sinusoidal congestion, necrosis of the parenchyma tissue, nuclear pyknosis, eosinophilic hepatocellular degeneration, pigment accumulation, an increase in the number and size of melanomacrophage centers. Liver tumors with severe chronic inflammation were occasionally found in fish at Al-Khadoud area (first-time report). The histological lesions were comparatively most severe in the liver. Despite heavy metals assessment did not show marked differences among sites, histopathological biomarkers indicated that the surveyed fish are living under stressful environmental conditions. So, we suggest use those biomarkers in future monitoring of aquatic systems.     

Relationship between arsenic accumulation in tissues and hematological parameters in mullet caught in Faro Lake: a preliminary study

The authors investigated the arsenic (As) accumulation in different tissues (muscle, gill, liver, stomach, and intestine) and the possible correlation between tissue concentration and hematological parameters in mullet (Mugil cephalus Linnaeus, 1758) caught in Faro Lake (Messina, Sicily, Italy). On all fish, hematological analyses of blood samples, measurement of biometric indices, and the removal of the muscles, gills, liver, stomach, and intestine for the determination of arsenic concentration were performed. A hemogram was performed to find effects of arsenic concentration in tissues on hematological variables. One-way analysis of variance showed significant differences of arsenic concentration in different tissues, with higher values in the gill. The correlation between hematological parameters and tissue arsenic concentration showed a statistical significance for red blood cell (RBC), hemoglobin concentration (Hb), and hematocrit (Hct) with the liver As concentration. Biometric indices (weight, length, and fork length) showed a significant correlation with As concentration of the muscle and liver also. Our results indicate the role of some hematological parameters as biomarkers useful to monitoring anthropogenic load of arsenic in water and sediment, because variations of these parameters represent one of the effects that arsenic exposure can have on fish.

     

The relationships between heavy metal (Cd,Cr, Cu,Fe, Pb,Zn) levels and the size of six Mediterranean fish species

Heavy metal (Cd, Cr, Cu, Fe, Pb, Zn) concentrations in the muscle, gill and liver of six fish species (Sparus auratus, Atherina hepsetus, Mugil cephalus, Trigla cuculus, Sardina pilchardus and Scomberesox saurus) from the northeast Mediterranean Sea were measured and the relationships between fish size (length and weight) and metal concentrations in the tissues were investigated by linear regression analysis. Metal concentrations (as microg/g d.w.) were highest in the liver, except for iron in the gill of Scomberesox sauris and lowest in the muscle of all the fish species. Highest concentrations of Cd (4.50), Cr (17.1) and Pb (41.2) were measured in liver tissues of T. cuculus, Sardina pilchardus and A. hepsetus, respectively. The liver of M. cephalus showed strikingly high Cu concentrations (202.8). The gill of Scomberesox saurus was the only tissue that showed highest (885.5) iron concentrations. Results of linear regression analysis showed that, except in a few cases, significant relationships between metal concentrations and fish size were negative. Highly significant (P<0.001) negative relationships were found between fish length and Cr concentrations in the liver of A. hepsetus and M. cephalus, and Cr concentrations in the gill of T. cuculus. Cr and Pb concentrations in the liver and Cu concentrations in all the tissues of Scomberesox saurus also showed very significant (P<0.001) negative relationships. Negative relationships found here were discussed.     

Development of a biotic ligand model (BLM) predicting nickel toxicity to barley (Hordeum vulgare)

A biotic ligand model (BLM) was developed to predict nickel toxicity, affecting root growth of barley (Hordeum vulgare), in nutrient solutions. The extent to which Ca(2+), Mg(2+), Na(+), K(+) ions and pH each influenced nickel toxicity was determined. Higher activities of Mg(2+) linearly increased the 4d EC50 Ni (2+) , while Ca(2+), Na(+), K(+) and H(+) activities did not significantly influence Ni(2+) toxicity. Stability constants for the binding of Ni(2+) and Mg(2+) to the biotic ligand were obtained: logK(NiBL)=5.27 and logK(MgBL)=3.47. Further, it was calculated that on average 57% of the biotic ligand sites needed to be occupied by nickel to induce 50% root growth inhibition. Auto-validation of the BLM indicated that predicted EC50s differed from the observed EC50s by a factor of less than 2, indicating that the BLM concept may also be used to predict metal toxicity to terrestrial plants.